LIBRARY Main Lib, Agric, Depfc UNIVERSITY OF CALIFORNIA. Deceive BIOLOGY Sl^/4 OGY Accessions No. 1 ' { (& b xviii LIST OF ILLUSTRATIONS. FIG. PACK 117. Colonies of Sternberg's Micrococcus, x 100 (Frankel and Pfeiffer) . 237 118. Acute Catarrhal Pneumonia, x 480 (Hamilton) > . . 239 119. Infectious Pleuro-pneumonia of Cattle (Hamilton) . . . 240 120. Infectious Pleuro-pneumonia of Cattle (Hamilton) . . .241 121. Bacillus of Influenza, x 1000 (Itzerott and Niemann) . . 248 122. Bacillus of Influenza, x 1200 (E.M.C) . 249 123. Bacilli of Plague and Phagocytes, x 800 (Aoyama) . . . 253 124. Spirillum Obermeieri in Blood of Monkey inoculated with Spirilla after Removal of the Spleen (Soudakewitch) . . . 258 125. Pure-cultivations of Streptococcus Pyogenes (E.M.C.) . 263 126. Free Surface of Diphtheritic Larynx, x 350 (Hamilton) . . 331 127. Bacillus of Diphtheria ; from a Cultivation on Blood Serum, x 1000 (Frankel and Pfeiffer) . . . . . . .332 128. Pure-cultures of Bacillus Diphtheria on Glycerine-Gelatine (E.M.C.) 333 129. Typhoid Fever. Ileum of Adult, showing Sloughy and Infiltrated Patches (Hamilton) . . . . . . .341 130. Typhoid Bacilli from a Colony on Nutrient Gelatine, x 1000 (Frankel and Pfeiffer) . . . . . . .342 131. Typhoid Bacilli, x 950 (Baumgarten) ..... 342 132. Flagella of Typhoid Bacilli, x 1000 (Frankel and Pfeiffer) . . 343 133. Colonies of the Typhoid Bacillus (Frankel and Pfeiffer) . . 344 134. Pure-culture of Typhoid Bacilli inoculated in the Depth of Nutrient Gelatine (Baumgarten) . . . . . . 344 135. Typhoid Bacilli in a Section of Spleen, x 800 (Fliigge) . . 345 136. Typhoid Bacilli in a Section of Intestine invading the Submucous and Muscular Layers, x 950 (Baumgarten) .... 346 137. Ulceration of the Intestine in a Typical Case of Swine-fever (E.M.C.) 348 138. Klein's Bacillus of Swine-fever (No. 1) . . . . 349 139. From a Preparation of Bronchial Mucus of Klein's Swine-fever Bacillus (No. 2) ....... 349 140. Bacilli from an Artificial Culture with Spores, Bacillus No. 2 (Klein) 349 141. Blood of Fresh Spleen of a Mouse after Inoculation with Swine-fever Bacillus No. 2 (Klein) 350 142. Bacilli of Swine Erysipelas (Baumgarten) .... 356 143. Blood of Pigeon inoculated with Bacilli of Swine Erysipelas, x 600 (Schutz) . . ... . . • .356 144. Pure-culture in Nutrient Gelatine of Bacilli from Swine Erysipelas (Baumgarten) . . . . . . . 357 145. Cover-glass Preparation of a Drop of Meat Infusion containing a Pure-cultivation of Comma-bacilli (Koch) . . . .361 146. Arthrospores of Comma-bacilli (Hueppe) .... 361 147. Flagella of Comma-bacilli ; stained by Loffler's Method (Frankel and Pfeiffer) . . . . . . . . :$62 148. Involution Forms of Comma-bacilli, x 700 (Van Ermengem) . 362 149. Colonies of Comma-bacilli on Nutrient Gelatine ; natural size (Koch) 362 150. Colonies of Koch's Comma-bacilli, x 60 (E.M.C.) . . . 363 151. Cover-glass Preparation from the Contents of a Cholera Intestine, x 600 (Koch) . . . . . . .363 152. Cover-glass Preparation of Cholera Dejecta on Damp Linen, x 600 (Koch) ......... 363 15.'!. Section of the Mucous Membrane of a Cholera Intestine, x 600 (Koch) . . . 364 LIST OF ILLUSTRATIONS. XIX FIG. PAGE 154. Pure-cultivations in Nutrient Gelatine of Koch's and of Finkler's Comma-bacilli (E.M.C.) ...... 365 155. Comma-shaped Organisms with other Bacteria in Sewage-con- taminated Water, x 1200 ...... 36(5 156. Comma-bacilli of the Mouth, x 700 (Van Ermengem) . . 367 157. Finkler's Comma-bacilli, from Cholera nostras, x 700 (Fliigge) . 367 158. Deneke's Comma-bacilli, from Cheese, x 700 (Fliigge) . . 367 159. Pure-cultivation of the Spirillum Finkler- Prior, in Nutrient Gelatine (E.M.C.) . . . . . ... .370 160. Tropical Dysentery ; Mucous Membrane of Large Intestine (Hamilton) 372 161. Tubercle of the Lung in a very Early Stage, x 400 (Hamilton) . 376 162. Primary Tubercle of Lung two to three weeks old, x 50 (Hamilton) 377 163. Large Oval Giant Cell from Tubercle of Lung, x 300 (Hamilton) . 377 164. Bacillus Tuberculosis, from Tubercular Sputum, x 2500 (E.M.C.) . 379 165. Pure-cultivation of the Tubercle Bacillus on Glycerine Agar-agar (E.M.C.) . 380 166. Pure-cultivation in Glycerine Agar-agar after ten months' growth (E.M.C.) . , ... . .381 167. Pure-cultivations of Tubercle Bacillus in Glycerine Agar-agar ; a sub- culture from a Pure-culture in Glycerine Milk (E.M.C.) . . 381 16S. Section through a Lupus Nodule of the Nose (Hamilton) . . 387 169. Tubercular Ulceration of Mucosa of Ileum (Hamilton) . . 388 170. Section of Lupus of the Skin ; Giant Cell containing Tubercle Bacillus (Fliigge) ....... 389 171. Tuberculosis of Pleura; "Grape-disease" (E.M.C.) . . .390 172. Tubercular Ulceration of the Intestine of a Cow (E.M.C.) . . 393 1 ::!. Tubercular Ulceration of the Intestine of a Rabbit (E.M.C.) . . 395 174. Tubercular Lungs of Rabbit (E.M.C.) . . . . .396 175. Cover-glass Preparation of Pus from a Chancre, x 1050 (Lustgarten) 410 17<). Wandering Cell containing Bacilli (Lustgarten) . . . 410 177. Section of Liver from a Case of Actinomycosis in Man (E.M.C.) . 417 178. Actinomycotic Tumour in the Throat of a Steer (E.M.C) . . 424 179. Actinomycotic Tumour of the Cheek (E.M.C.) .... 424 180. Steer with Emaciation the Result of Actinomycosis (E.M.C.) . 425 181. Actinomycotic Growths from the Pleura resembling " Grape- Disease" (E.M.C.) . . . . . .425 182. Actinomycosis of the Skin (E.M.C.) . . . . .430 is:!. Part of Human Foot with Madura Disease (E.M.C.) . . .448 1S4. Bacilli of Glanders, x 700 (Fliigge) . . . . .452 185. Section of a Branch of the Pulmonary Artery showing Glanders Bacilli penetrating the Wall (Hamilton) .... 453 186. Pure-culture of the Tetanus Bacillus in Grape-sugar Gelatine (Frankel and Pfeiffer) . . . . . . . .458 187. Foot of Sheep showing Disease of Horn (Brown) . . . 465 188. Section through the Foot showing a Crack extending through the Wall 465 189. Secreting Membrane covered with Fungoid Growths (Brown) . 466 190. Advanced Form of Disease of Skin between the Claws (Brown) . 466 191. Distortion of Hoof in an Advanced Form of Foot-rot (Brown) . 467 192. Diseased Comb (Cowan) . . . . . .469 193. Spores of Bacillus Alvei (E.M.C.) . . . . .470 191. Pure-culture in Nutrient Gelatine (Cheshire and Cheyne) . . 470 XX LIST OF ILLUSTRATIONS. *-:u. rAOK 195. Cultivation on the Surface of Gelatine (Cheshire and Cheyne) . 471 196. Cladothrix Dichotoma (Zopf) . . . 479 197. Friedlander's Pneumococcus, x 1500 (Zopf) . . . 483 198. Ascococcus Billrothh (Cohn) . . 498 199. Clostridium Butyricum (Prazmowski) . . . . . 503 200. Bacillus Cyanogenus, x 650 (Neelsen) .... 507 "201. Pure-cultivation of Bacillus figurans on the Surface of Nutrient Agar-agar (E.M.C.) . . . 509 202. Photograph of Part of an Impression Preparation of Bacillus figurans on Nutrient Gelatine, x 50 (E.M.C.) . . . 510 203. Part of the same Specimen, x 200 . . . . .510 204. Bacillus Indicus : Colonies in Agar, x 60 . . . .518 205. Bacillus Neapolitans, x 700 (Emmerich) .... 522 206. Bacillus Megatherium (De Bary) . . . . .524 207. Pure-culture of Bacillus Megatherium in Gelatine (E.M.C.) . . 524 208. Bacillus Putrificus Coli, x 1000 (Bienstock) .... 529 209. Bacillus Pyogenes Foetidus, x 790 (Passet) .... 530 210. Bacillus Saprogenes, No. 1 (Rosenbach) . . 531 211. Bacillus Subtilis with Spores (Baumgarten) .... 535 212. Pure-culture of Bacillus Subtilis in Nutrient Gelatine (Baumgarten) . 535 213. Pure-culture of Bacillus Subtilis on the Surface of Nutrient Agar (E.M.C) . . 536 214. Bacterium Zopfii (Kurth) ...... 542 215. Beggiatoa Alba (Zopf) ..... .543 216. Phase-forms of Beggiatoa Persicina (Warming) . . . 544 217. Cladothrix Dichotoma (Zopf) ...... 545 218. Crenothrix Kuhniana (Zopf) ...... 546 219. Leuconostoc Mesenteroides (Van Tieghem and Cienkowski) . . 550 220. Micrococcus in Pyaemia in Babbits (Koch) .... 556 221. Proteus Mirabilis ; Swarming Islands on the Surface of Gelatine, x 285 (Hauser) 5(51 222. Proteus Mirabilis ; Involution Forms, x 524 (Hauser) . . 5(51 223. Proteus Vulgaris, x 285 (Hauser^ . . . . .562 224. Sarcina, x 600 (Fliigge) ...... 5(53 225. Spirochteta Plicatile (E.M.C.) . . . . . . 566 226. Comma-bacilli in Water contaminated with Sewage . . . 567 227. Comma-bacilli of the Mouth, x 700 (Van Ermengem) . . 568 228. Deneke's Comma-bacilli, from Cheese, x 700 (Fliigge) . . 568 229. Streptococcus in Progressive Tissue Necrosis in Mice (Koch) . 571 230. Vibrio Rugula, x 1020 (Prazmowski) . . . . .574 231. Black Torula; Pure-cultivation on Potato (E.M.C.) . . . 581 232. Head and Neck of Calf with Advanced Ringworm (Brown) . . 585 233. Non-pigmented Amoeboid Forms (Marchiafava and Celli) . . 590 234. Pigmented Amoeboid Forms (Golgi) ..... 590 235. Semi-lunar Bodies of Laveran (Golgi) ... . 590 236. Rosette Forms with Segmentation (Golgi) . . . 591 237. Flagellated Forms (Vandyke Carter) . . . . .591 238. " Surra " Parasites, occurring Singly and Fused, x 1200 . . 598 239. Parasites in the Blood of Rats (Lewis) .... 599 240. A Monad in Rat's Blood, x 3000 (E.M.C.) .... 601 241. Monads in Rat's Blood, x 1200 (E.M.C.) .... 602 242. Monads in Rat's Blood stained with Methyl Violet, x 1200 (E.M.C.) 603 LIST OF ILLUSTRATIONS. XXI HO. I'AGK 243. Organisms in the Blood of Mud-fish (Mitrophanow) . . . 604 244. Organisms in the Blood of the Carp (Mitrophanow) . . . 605 245. Amoeba cola in Intestinal Mucus (Losch) .... 611 2 It;. Warm Stage (Schafer) . . . . . . .613 247. Warm Stage (Strieker) ... . 614 IMS. Combined Gas Chamber and Warm Stage (Strieker) . . . 614 249. Vertical Micro-photographic Apparatus (Leitz) . . . 622 250. Koch's Steam Steriliser (Muencke) ..... 623 251. Hot-air Steriliser (Muencke) ...... 623 252. Section of Hot-air Steriliser (Muencke) . . 623 253. Hot-water Filtering Apparatus with Ring Burner (Rohrbeck) . 624 254. Wire-cage for Test-tubes (Muencke) , 626 255. Platinum-needles (E.M.C.) ...... 626 256. Damp Chamber for Plate-cultivations (E.M.C.) . . .626 257. Apparatus for Plate cultivations (E.M.C.) . . .627 253. Box for Glass-plates (Muencke) . . .627 259. Glass Benches for Glass-plates (Becker) .... 627 260. Israel's Case (Becker) . . . 628 261. Damp Chamber for Potato Cultivations (E.M.C.) . . 628 262. Koch's Serum Steriliser (Muencke) . . 629- 263. Serum Inspissator (Muencke) .... . 62D 264. D'Arsonval's Incubator (Muencke) , 631 265. Schlosing's Membrane Regulator (Muencke) .... 632 266. Gas Burner protected with Mica Cylinder (Muencke) . 633 267. Koch's Safety Burner (Muencke) ..... 633 268. Babes' Incubator (Muencke) . . . .634 269. Moitessier's Gas-pressure Regulator (Muencke) . . . 634 270. Reichert's Thermo-regulator (Muencke) . . 635 271. Meyer's Thermo-regulator (Muencke) ..... 636 272. Siphon Bottle with Flexible Tube, Glass Nozzle, and a Mohr's Pinch- cock (E.M.C.) .637 273. Desiccator (E.M.C.) . . . . . . .638 DESCRIPTION OP PLATES. DESCRIPTION OF PLATE I. Bacteria, Schizomycetes, or Fission Fungi. Following p. 14. 1. Cocci singly and varying in size. 2. Cocci in chains or rosaries (strepto- coccus). 3. Cocci in a mass (staphylococcus). 4 and 5. Cocci in pairs (diplococcus). 6. Cocci in groups of four (merismopedia). 7. Cocci in packets (sarcina). 8. Bacterium termo. 9. Bacterium termo x 4000 (Dallinger.and Drysdale). 10. Bacterium septiccemice hcemorrhagicce . 11. Bacterium pneu- monice crouposce. 12. Bacillus subtilis. 13. Bacillus murisepticus. 14. Bacillus diphtheria. 15. Bacillus typhosus (Eberth). 16. Spirillum undula (Cohn). 17. Spirillum volutans (Cohn). 18. Spirillum cholera Asiaticce. 19. Spirillum Obermeieri (Koch). 20. Spirochceta plicatilis (Fliigge). 21. Vibrio rugula (Prazmowski). 22. Cladothrix Forsteri (Cohn). 23. Cladothrix dichotoma (Cohn). 24. Nonas OJtenii (Cohn). 25. Nonas Warmingii (Cohn). 26. Rhabdomonas rosea (Cohn). 27. Spore-formation (Bacillus alvei). 28. Spore-formation (Bacillus anthracis). 29. Spore-formation in bacilli cultivated from a rotten melon (Frankel and Pfeiffer). .30. Spore-formation in bacilli cultivated from earth (Frankel and Pfeiffer). 31. Involution-form of Crenothrix (Zopf). 32. Involution-forms of Vibrio serpem (Warming). 33. Involution- forms of Vibrio rugula (Warming). 34. Involution-forms of Clostridium polymyxa (after Prazmowski). 35. Involution-forms of Spirillum cholera AsiaticfB. 36. Involution-forms of Bacterium aceti (Zopf and Hansen). 37. Spirulina-form of Beggiatoa alba (Zopf). 38. Various thread-forms of Bacterium merismopedioides (Zopf). 39. False-branching of Cladothrix^Zopf). DESCRIPTION OF PLATE II. Pure-cultivations of Bacteria. Following p. 100. FIG. 1. — In the depth of Nutrient Gelatine. A pure-cultivation of Koch's comma-bacillus (Spirillum choleras Asiaticae) showing in the track of the needle a funnel-shaped area of liquefaction enclosing an air-bubble, and a white thread. Similar appearances are produced in cultivations of the comma-bacillus of Metchnikoff. FIG. 2. — On tlie surface of Nutrient Gelatine. A pure-cultivation of Bacillus typhosus on the surface of obliquely solidified nutrient gelatine. xxii DESCRIPTION OF PLATES. xxiii FIG. 3. — On the surface of Nutrient Agar-agar. Pure-cultivation of Bacillus indicus on the surface of obliquely solidified nutrient agar-agar. The growth has the colour of red sealing-wax, and a peculiar crinkled appearance. After some days it loses its bright colour and becomes purplish, like an old cultivation of Micrococcus prodigiosus. FIG. 4. — On the surface of Nutrient Agar-agar. A pure-cultivation obtained from an abscess (Staphylococcus pyogenes aureus). FIG. 5.— On the surface of Nutrient Agar-agar. A pure-cultivation obtained from green pus (Bacillus pyocyaneus). The growth forms a whitish, transparent layer, composed of slender bacilli, and the green pigment is diffused throughout the nutrient jelly. The growth appears green by transmitted light, owing to the colour of the jelly behind it. FIG. (\.-On tlie surface of Potato. A pure-cultivation of the bacillus of glanders on the surface of sterilised potato. DESCRIPTION OF PLATE III. Plate-cultivation. Following p. 108. This represents the appearance of a plate-cultivation of the comma-bacillus of Cholera nostras, when it is examined over a slab of blackened plate-glass. The drawing was made from a typical result of thinning out the colonies by the process of plate-cultivation. At this stage they were completely isolated one from the other ; but later they became confluent, and produced complete liquefaction of the gelatine. DESCRIPTION OF PLATE IV. Streptococcus Pyogenes. Following p. 178. FIG. 1. — From a cover-glass preparation of pus from a pyaemic abscess. Stained with gentian-violet by the method of Gram, and contrast-stained with eosin. x 1200. Powell and Lealand's apochromatic T\ Horn. imm. E. P. 10. FIG. 2. — From cover-glass preparations of artificial cultivations of the strepto- coccus in broth and in milk at different stages of growth, x 1200. Powell and Lealand's apocbromatic -fa Horn. imm. E. P. 10. In these preparations there is a great diversity in size and form of the chains and their component elements. In the drawing examples are figured of the following: (a) Branched chains. (b) Simple chains composed of elements much smaller than the average size. O) Chains with spherical and spindle-shaped elements at irregular intervals. These are conspicuous by their size, and are sometimes terminal. (d e) Chains in which the elements are more or less uniform in size. (/) Complex chains with elements dividing both longitudinally and transversely, and varying considerably in size in different lengths of the same chain. XXiv DESCRIPTION OF PLATES. DESCRIPTION OF PLATE V. Bacillus Anthracis. Following p. 192. FIG. 1. — From a cover-glass preparation of blood from the spleen of a guinea- pig inoculated with blood from a sow. x 1200. Powell and Lealand's apochromatic T\ Horn. imm. E. P. 10. FIG. 2. — From a section of a kidney of a mouse. Under a low power the preparation has exactly the appearance of an injected specimen. Under higher amplification the bacilli are seen to have threaded their way along the capillaries between the tubules, and to have collected in masses in the glomeruli. Stained with Gram's method (gentian-violet), and eosin. x 500. FIG. 3. — Bacillus anthracis and Microcoocus tetragenits. From a section from the lungs of a mouse which had been inoculated with anthrax three days after inoculation with Mlcrococcus tetragenus. A double or mixed infection resulted. Anthrax-bacilli occurred in vast numbers, completely filling the small vessels and capillaries, and in addition there were great numbers of tetrads. Stained by Gram's method (gentian- violet), and with eosin. x 500. DESCRIPTION OF PLATE VI. f Bacillus Murisepticus. Following p. 224. FIG. 1. — From a section of a kidney of a mouse which had died after inocula tion with a pure-cultivation of the bacillus. With moderate amplification, the white blood-corpuscles have a granular appearance, and irregular granular masses are scattered between the kidney tubules. Stained by Gram's method with eosin. x 200. FIG. 2. — Part of the same preparation with high amplification. The granular appearances are found to be due to the presence of great numbers of extremely minute bacilli, x 1500. DESCRIPTION OF PLATE VII. Casual Cow-pox. Following p. 278. FIG. 1. — Case of W. P , a milker, infected from the teats of a cow with natural cow-pox. There was a large depressed vesicle with a small central crust and a tumid margin, the whole being surrounded by a well-marked areola and considerable surrounding induration. FIG. 2. — The same case a week later, showing a reddish-brown crust on a reddened elevated and indurated base. DESCRIPTION OF PLATES. XXV DESCRIPTION OF PLATE VIII. Bacillus diphtherias and Bacillus typhosus. Following p. 382. Kid. 1. — Cover-glass preparation from a pure-cultivation of Bacillus diph- therias on blood serum ; obtained from the throat in a typical case of diphtheria. Stained with gentian-violet, x 1200. FIG. 2. — Cover-glass preparation from a pure-cultivation of Bacillus typhosus on nutri,ent-agar ; from the spleen in a case of typhoid fever, Stained with gentian-violet, x 1200. DESCRIPTION OF PLATES IX. AND X. Swine Fever. Following p. 348. PLATE IX. — Part of intestine from a typical case of swine fever, showing scattered ulcers and ulceration of the ileo-csecal valve. PLATE X. — From the same case of swine fever. The lungs were extensively inflamed and partly consolidated, and the lymphatic glands were enlarged and of a deep reel or reddish-purple colour. DESCRIPTION OF PLATE XI. Bacillus tuberculosis. Following p. 378. The figures in this plate represent the bacilli of tuberculosis in different animals, examined under the same conditions of amplifica- tion and illumination, x 1200. Lamp-light illumination. FIG. 1. — Bacilli in pus from the wall of a human tubercular cavity. In this specimen the bacilli are shorter than those in tubercular sputum, and are very markedly beaded. FIG. 2. — Bacilli in pus from a tubercular cavity from another case in man. They are present in the preparation in enormous numbers. The pioto- plasm occupies almost the whole of the sheath, and the bacilli are strikingly thin and long. FIG. 8.— Bacilli in sputum from an advanced case of phthisis, showing the ordinary appearance of bacilli in sputum ; some beaded, others stained in their entirety; occurring both singly and in pairs, and in groups resembling Chinese letters. FIG. 4. — Bacilli in a section from the lung in a case of tuberculosis in man. The bacilli in human tuberculosis are found in, and between, the tissue cells ; and sometimes, as in equine and bovine tuberculosis, in the interior of giant cells, but not so commonly. FIG. 5. — From a cover-glass preparation of the deposit in a sample of milk from a tubercular cow. The bacilli were longer than the average length of bacilli in bovine tissue sections, and many were markedly beaded. XXvi DESCRIPTION OF PLATES. FIG. (5. — From a section of the brain in a case of tubercular meningitis in a calf, showing a giant cell containing bacilli with the characters usually- found in sections of bovine tuberculosis. FIG. 7. — From a section of the liver of a pig with tubercle bacilli at the margin of a caseous nodule. FIG. 8. — From a cover-glass preparation of a crushed caseous mesenteric gland from a rabbit infected by ingestion of milk from a cow with tuberculosis of the udder. FIG. 9. — From a section of lung in a case of equine tuberculosis, showing a giant cell crowded with tubercle bacilli. FIG. 10. — From a section of lung from a case of tuberculosis in the cat, with very numerous tubercle bacilli. FIG. 11. — From a cover-glass preparation of a crushed caseous nodule from the liver of a fowl, with masses of bacilli. These are for the most part short, straight rods ; but other forms, varying from long rods to mere granules, are also found. FIG. 12. — From sections of the liver and of the lung in a case of tubercu- losis of a Khea. Isolated bacilli are found, as well as bacilli packed in large cells, colonies of sinuous bacilli, and very long forms with terminal spore-like bodies and free oval grains. The preparations from which these figures were drawn were all stained by the Ziehl-Neelsen method, with the exception of the first, which was stained by Ehrlich's method. DESCRIPTION OF PLATE XII. Tubercular Mammitis. Following p. 394. FIG. 1.— From a section of the udder of a milch cow. The tubercular deposit is seen to invade the lobules of the gland. Lobules comparatively healthy are marked off, more or less sharply, from the diseased ones in which the new growth in its progress compresses and obliterates the alveoli. Stained by the Ziehl-Neelsen method and with methylene-blue. x 50. FIG. 2. — Part of the same preparation. On the right of the section part of a healthy lobule is seen. On the left a lobule is invaded by tubercular new growth composed of round cells, epithelioid cells and typical giant cells. Tubercle bacilli can be seen both singly and collected in groups. They are found in and between the cells, and in the interior of giant cells. Bacilli may be seen between the cells lining an alveolus and projecting into its lumen, x 800. DESCRIPTION OF PLATE XIII. Tuberculosis in Swine. Following p. 400. Section of liver of a pig with scattered tubercular nodules. Microscopical sections of the liver showed tubercle bacilli in very small numbers. DESCRIPTION OF PLATES. XXvii DESCRIPTION OF PLATE XIV. Bacillus Leprae. Following p. 408. FIG. 1. — From a section of the skin of a leper. The section is, almost in its entirety, stained red, and, with moderate amplification, has a finely granular appearance. Stained by the Ziehl-Neelsen method (carbolised fuchsine and methylene-blue). x 200. FIG. 2. — Part of the same preparation with high amplification, showing that the appearances described above are due entirely to an invasion of the tissue by the bacilli of leprosy, x 1500. DESCRIPTION OF PLATES XV. AND XVI. Actinomyces. Following p. 432. PLATE XV. FIG. 1.— From a preparation of the grains from an actinomycotic abscess in a boy ; examined in glycerine. The drawing has been made of a com- plete rosette examined by focussing successively the central and peripheral portions. Towards the centre the extremities of the clubs are alone visible ; they vary in size, and if pressed upon by the cover-glass give the appearance of an irregular mosaic. Towards the periphery the clubs are seen in profile, and their characteristic form recognised. At one part there are several elongated elements, composed of separate links, x 1200. FIG. 2. — Different forms of clubs from preparations in which the rosettes have been flattened out by gentle pressure on the cover-glass, x 2500. (a) Single club. (ft) Bifid club, (c) Club giving rise to four secondary clubs. (cT) Four clubs connected together, recalling the form of a bunch of bananas. ( tPf^MlOd AOSWCsTS ^ndl lOB* CCB^tstMTl HI if sffEmffP|W ^IflffBttfv AO<6? preparations are left for one minute in solution of tannin, washed in dirtilipd water, transferred for a minute to 50 per cent, phospho- molybdic acid, again washed and stained from three to five 92 BACTERIOLOGY. in hot saturated solution of f uchsine in aniline water, washed in watery dried on filter paper, and mounted in balsam. The tannin solution consists of 1 part of tannin to 100 cc. of 50 per cent, alcohol. Nicolle and Morax also, have modified Loffler's method. Per- fectly clean cover-glasses are used, and the film is dried without fixing in the flame. Cover-glasses are covered with the mordant, and heated for about ten seconds, and when steam rises the mordant is shaken off and the film rinsed with water. The same process is repeated three or four times, and finally the cover-glass is stained with Neelsen's solution, holding it over the flame once or twice for a quarter of a minute ; it is then washed and examined. Bunge prefers as a mordant a mixture of aqueous solution of tannin with 1 in 20 aqueous solution of sesquichloride of iron in the proportion of 3 parts of the tannin solution, 1 part of the iron solution, with the addition of 1 cc. of a saturated watery solution of f uchsine added to 10 cc. of the mixture. The mordant is kept before use, and applied for five minutes. The preparation is then washed and stained with ISTeelsen's solution. In another plan the cover-glasses are immersed for one half to one minute in 5 per cent, solution of acetic acid, washed and dried. The mordant is then applied three or four times, and the cover-glasses washed, dried, and then stained with gentian-violet, dipped in 1 per cent, acetic acid, washed, dried, and mounted. Peroxide of hydrogen may be added to the mordant r drop by drop ; it becomes reddish-brown in colour, and must be shaken up and filtered before use. Cover-glasses are exposed to its action for about a minute, and Neelsen's solution is used for staining. Hessert dispenses with the mordant. The film is fixed by treating cover-glasses with a saturated alcoholic solution of corrosive sublimate. .After washing, the cover-glass is stained for thirty to forty minutes in a hot dye, by preference a 10 per cent, watery solution of saturated alcoholic solution of fuchsine. COVER-GLASS IMPRESSIONS. One of the most instructive methods for examining micro- organisms is to make an impression-penetration. This enables us, in many cases, to study the relative position of individual micro- organisms one to another in their growth on solid cultivating media, and in some cases produces the most exquisite preparations for the microscope. A perfectly clean, usually small-sized, cover-glass is carefully deposited on a plate-cultivation, and gently and evenly pressed down. One edge is then carefully levered up, with a needle, MICROSCOPICAL EXAMINATION OF BACTERIA. 93 and the cover-glass lifted off by means of forceps. It is then allowed to dry, passed through the flame three times, and stained as already described. In some cases of plate- cultures, especially where no liquefaction has taken place, the growth is bodily trans- ferred to the cover-glass, and a vacant area left on the gelatine or agar-agar, corresponding exactly with the form and size of the cover-glass employed. PRESERVATION OF PREPARATIONS. After examining a cover-glass preparation with an oil immersion objective the cedar oil must be carefully wiped off, and the slide set aside for the Canada balsam to set. At a convenient time all preparations should be sealed with a ring of Hollis' glue ; the cedar oil used at subsequent examinations of the specimen will not be able to work its way under the cover-glass, and prevent the balsam from hardening. When it is ringed cedar oil can be readily wiped off, and the specimen cleaned without danger of moving the cover-glass and injuring the preparation. (B) BACTERIA IN SECTIONS OF TISSUES. Methods of Hardening and Decalcifying Tissues. — To harden small organs, such as the viscera of a mouse, they should be placed on a piece of filter- paper at the bottom of a small wide -mouthed glass jar, and covered with about twenty times their volume of absolute alcohol. Larger organs, pathological growths, etc., are treated in the same way, but must first be cut into small pieces, or cubes, varying from a quarter of an inch to an inch in size. Miiller's fluid may also be employed, and methylated spirit may be sub- stituted for alcohol, from motives of economy. Tissues hardened in absolute alcohol are ready for cutting in two or three days, and those hardened in Miiller's fluid in as many weeks. Teeth, or osseous structures, must first be placed in a decalcifying solution, such as Kleinenberg's. When sufficiently softened, they are allowed to soak in water, to wash out the picric acid, and then transferred through weak spirit to absolute alcohol. Ebner's solu- tion also gives excellent results, especially when the structures to be decalcified are placed in fresh solution from time to time. Methods of Embedding, Fixing, and Cutting. — The author finds that freezing with ether combined with the method of em- bedding in celloidin gives excellent results. The pieces of tissue to be embedded are placed, after the process of hardening is com- 94 BACTERIOLOGY, pleted, in a mixture of ether and alcohol for an hour or more. They are then transferred to a solution of celloiclin in equal parts of ether and alcohol, and left there, usually for several hours. The piece of tissue is then placed in a glass capsule, and some of the celloidin solution poured over it. The capsule can be placed FIG. 27. — SWIFT'S FREEZING MICROTOME. bodily in 60 to 80 per cent, alcohol, and left until the following morning. The celloidin will then be of the consistency of wax. The piece of tissue is next cut out, and after trimming off superfluous celloidin is put in water until it sinks. It is then transferred to gum, and frozen and cut with a freezing microtome. For cutting with Jung's microtome, the tissues are embedded MICROSCOPICAL EXAMINATION OF BACTERIA. 95 in paraffine or celloidin, and mounted on cork ; or, if firm enough, they may be fixed upon cork without any embedding material at all. Paraffine, dissolved in chloroform, will be found very service- able as an embedding material. Corks ready cut for the clamp of the microtome are smeared over with the solution of celloidin. This can be applied with a glass rod to the surface which is to receive the piece of tissue- The corks are then set aside for the film of celloidin to harden. In the case of lung, or degenerated broken-down tissue, the specimen should be left for a much longer time than is found to be sufficient for firmer structures. When ready, it is removed from the celloidin solution with forceps and placed upon the pre- FIG. 28. — JUNG'S MICROTOME. pared cork. Enough of the solution, which is of syrupy consistence, is allowed to fall on the piece of tissue to cover it completely, and the mounted specimen is placed in the alcohol to harden. The specimen will be ready for cutting next day. The specimen may be more neatly embedded by fixing it with a pin in a small paper tray, pouring the celloidin solution over it, and then placing the tray in alcohol to harden the celloidin. The embedded specimen is then fixed on a cork, which has been cut for the clamp of the microtome. The celloidin in the section disappears in the process of clearing with clove-oil. In the case of specimens embedded in celloidin, or mounted directly on a cork, the tissue, as well as the blade of the knife, should be kept constantly bathed with alcohol, and the sections transferred from the blade with a camel's-hair brush, and floated in alcohol. 96 BACTERIOLOGY. For fixing directly on cork, small organs and pieces of firm tissue such as the kidneys of a mouse, or liver, we may employ gelatine or glycerine gelatine, liquefied over a Bunsen burner in a porcelain capsule. Glycerine gelatine may be used with advantage for fixing irregular pieces of tissue, as it does not become of a consistency that would injure the edge of the knife. The cork, with specimen affixed, is placed in alcohol, and is ready for cutting sections next day. Material infiltrated with paraffine must be cut perfectly dry, and the sections prevented from rolling up by gentle manipulation with a camel's-hair brush. They must then be picked off the blade of the knife with a clean needle, and dropped into a watch-glass containing xylol. This dissolves out the paraffine. The sections are then transferred to alcohol to get rid of the xylol, and then to the staining solution. Staining Bacteria in Tissue Sections, — Sections of fresh tissues made with the freezing microtome are to be floated in '8 per cent, salt solution, and then carefully transferred, well spread out on a platinum lifter, to a watch-glass containing absolute alcohol. Simi- larly, sections selected from those cut with Jung's microtome may be transferred from the spirit to absolute alcohol. The sections may be then stained by any of the methods to be described. It is often advisable to employ some method which will enable one to study the structure of the tissue itself ; and sections, however stained, should always be first examined with low powers, to enable one to recognise the tissue under examination, and to examine in many cases the topographical distribution of masses of bacteria. With a power of about 250 diams. (one-sixth), very many bacteria can be distinguished ; and with the oil immersion lenses the minutest bacilli and micrococci can be recognised, and the exact form of individual bacteria accurately determined. As most good modern instruments are provided with a triple nose-piece, there is no loss of time in examining a preparation successively with these different powers. Weigert's Method. — A very useful method for staining both the tissue and the bacteria is as follows : Place the sections for from six to eighteen hours in a 1 per cent, watery solution of any of the basic aniline dyes (methyl violet, gentian violet, fuchsine, Bis- marck brown). To hasten the process, place the capsule containing the solution in the incubator, or heat it to 45° C. A stronger solution may also be employed, in which case the sections are far more rapidly stained, and are easily over-stained. In the latter case MICROSCOPICAL EXAMINATION OF BACTERIA. 97 they must be treated with a half-saturated solution of carbonate of potash. In either case the sections are next washed with distilled water, and passed through 60 per cent, alcohol into absolute alcohol. When almost decolorised, spread out the section carefully on a platinum lifter and transfer it to clove-oil, or stain with picro-carmine solution (Weigert's) for half an hour, wash in water, alcohol, and then treat with clove-oil. After the final treatment with clove- oil, transfer with the platinum lifter to a clean glass slide. Dry the preparation by pressure with a piece of filter-paper folded several times, and preserve in Canada balsam, dissolved in xylol. Gram's Method. — In the method of Gram sections are stained for ten minutes in a capsule containing aniline-gentian- violet solution. Great care must be taken not to injure the sections. If there is any difficulty in finding them, it is best to carefully pour off the stain and fill up the capsule with water. The sections are then readily visible, and can be taken up on the end of a glass rod and placed in the iodine and iodide of potassium solution, where they remain for two or three minutes, until stained uniformly brown and resembling in appearance a boiled tea-leaf. They are then placed in absolute alcohol, and washed by carefully moving the sections in the liquid with a glass rod. When completely decolorised they are spread out on a lifter, and transferred to clove-oil until completely clarified. Each is transferred with a lifter to a slide, and the clove-oil is run off and then completely removed by gently pressing two or three layers of filter-paper upon the section. Finally, the section is mounted in Canada balsam. The process of decolorisation may be hastened by transferring the section from alcohol to clove-oil, and back again to alcohol, repeating this two or three times. On examination the tissue appears colourless, or slightly tinged yellow from too long immersion in the iodine solution, while the micro-organisms are stained blue or blue-black. Double staining is obtained by transferring the sections after decolorisation to eosin, Bismarck brown, or vesuvin. They are left in a watery solution for two or three minutes, then again washed in alcohol, before clarifying in clove-oil and mounting in balsam. Another instructive method is to place the decolorised sections in picro-carminate of ammonia for three or four minutes, and then treat with alcohol and clove-oil. A similar result is obtained by placing the sections in Orth's solution (picro-lithium carmine), transferring to acidulated alcohol, and then passing through clove-oil and mounting in balsam. 7 98 BACTERIOLOGY. In Ehrlich's method delicate sections are liable to be injured by immersion in the nitric acid, and therefore Watson- Oheyne suggested the use of formic acid. ' The Ziehl-Neelsen method, in which sulphuric acid is used instead of nitric acid, is much to be preferred to Ehrlich's method. Ziehl-Neelsen Method. — The solution is warmed, and sections left in it for ten minutes. The red colour, which disappears when the section is placed in weak sulphuric acid (25 per cent.), may partly return when the section is placed in water. In this case the section must be again immersed in acid and passed backwards and forwards from acid to water until the red colour has completely, or almost completely, disappeared. It must be thoroughly washed in water to remove all traces of the acid, and then placed in a watery solution of methylene blue for two or three minutes, washed again in water, immersed in alcohol, clarified in clove-oil, and mounted in the usual way. Sections are brilliantly stained, and the results are very permanent. Many special methods of staining have been introduced, and will be given in subsequent chapters with the description of the bacteria to which they apply. The methods already described are those which are more or less in constant use in studying bacteria and in conducting original researches. CHAPTER IX. PREPARATION OF NUTRIENT MEDIA AND METHODS OF CULTIVATION. To cultivate micro-organisms artificially, and, in the case of the pathogenic bacteria, to fulfil the second of Koch's postulates, they must be supplied with nutrient material free from pre-existing micro-organisms'. Hitherto various kinds of nutrient liquids have been employed, and in many cases they still continue to be used with advantage, but for general use they have been, in a great measure, supplanted by the methods of cultivation on sterile solid media about to be described. The advantages of the latter methods are numerous. In the first place, in the case of liquid media, in spite of elaborate precautions and the expenditure of much labour and time, it was almost impossible or extremely difficult to obtain a pure culture. When a drop of liquid containing several kinds of bacteria is introduced into a liquid medium, we have a mixed cultivation from the very first. If in the struggle for existence some bacteria were unable to develop in the presence of others, or a change of temperature and soil allowed one form to predominate over another, then we might be led to the conclusion that many bacteria were but developmental forms of one and the same micro- organism ; while possibly the contamination of such cultures might lead to the belief in the transformation of a harmless into a patho- genic bacterium. The secret of the success of Koch's methods greatly depends upon the possibility, in the case of starting with a mixture of micro-organisms, of being able to isolate them completely one from another, and to obtain an absolutely pure growth of each cultivable species. When sterile nutrient gelatine has been liquefied in a tube and inoculated with a mixture of bacteria in such a way that the individual micro-organisms are distributed throughout it, and the liquid is poured out on a plate of glass and allowed to solidify, the individual bacteria, instead of moving about freely as in a liquid medium, are fixed in one spot, where they develop individuals of 99 DESCRIPTION OF PLATE II. Pure-cultivations of Bacteria. FIG. 1. — In the depth of Nutrient Gelatine. A pure-cultivation of Kochs comma-bacillus (Spirillum choleras Asiatics) showing in the track of the needle a funnel-shaped area of liquefaction enclosing an air-bubble, and a white thread. Similar appearances are produced in cultivations of the comma-bacillus of Metchnikoff. FIG. 2. — On the surface of Nutrient Gelatine. A pure-cultivation of Bacillus typhosus on the surface of obliquely solidified nutrient gelatine. FIG. 3. — On the surface of Nutrient Agar-agar. • Pure-cultivation of Bacillus indicus on the surface of obliquely solidified nutrient agar-agar. The growth has the colour of red sealing-wax, and a peculiar crinkled appearance. After some days it loses its bright colour and becomes purplish, like an old cultivation of Micrococcus prodigiosus. FIG. 4. — On the surface of Nutrient Agar-agar. A pure-cultivation obtained from an abscess (Staphylococcus pyogenes aureus). FIG. 5. — On tJie surface of Nutrient Agar-agar. A pure-cultivation obtained from green pus (Bacillus pyocyaneus). The growth forms a whitish, transparent layer, composed of slender bacilli, and the green pigment is diffused throughout the nutrient jelly. The growth appears green by transmitted light, owing to the colour of the jelly behind it. FIG. (5. — On the surface of Potato. A pure-cultivation of the bacillus of glanders on the surface of sterilised potato. Plate TL CO •o CO (N •00 OF THB TJNIVERSITY NUTRIENT MEDIA AND METHODS OF CULTIVATION. 101 squeezing through a linen cloth or a meat press. The red juice thus obtained must be brought up to a litre by transferring it to a large measuring glass and adding distilled water. It is then poured into a sufficiently large and strong beaker, and set aside after the addition of 10 grammes of peptone, 5 grammes of common salt and 100 grammes of best gelatine. In about half an hour the gelatine is sufficiently softened, and subsequent heating in a water-bath causes it to be completely FIG. 30.— HOT Am STERILISKK. dissolved. The danger of breaking the beaker may be avoided by placing a cloth, several times folded, at the bottom of the water-bath. The next process requires the greatest care and attention. Some micro-organisms grow best in a slightly acid, others in a neutral or slightly alkaline, medium. For example, for the growth and characteristic appearances of the comma bacillus of Asiatic cholera a faintly alkaline soil is absolutely essential. This slightly alkaline medium will be found to answer best for most micro-organisms, and may be obtained as follows : — With a clean glass rod dipped in the mixture, the reaction upon litmus- paper may be ascertained, and a concentrated solution of carbonate of soda must be added drop by drop, until red litmus- 102 BACTERIOLOGY. paper becomes faintly blue. If it has been made too alkaline, it can be neutralised by the addition of lactic acid. Finally, the mixture is heated for an hour in the water-bath. Ten minutes before the boiling is completed, the white of an egg beaten up with the shell is added, and the liquid is then filtered while hot. For the filtration, the hot- water apparatus (Fig. 31) can be used with advantage, furnished with a filter of Swedish paper, which may be conveniently made in the following way : — About eighteen inches square of the best and stoutest filter paper is first folded in the middle, 'and then creased into sixteen folds. The filter is made to fit the glass funnel by gathering up the folds like a fan, and cutting off the superfluous part. The creasing of each fold should be made firmly to within half an inch of the apex of the filter, which part is to be gently inserted into the tube of the funnel. To avoid bursting the filter at the point, the broth, when poured out from the flask, should be diiected against the side of the filter with a glass rod. During filtration the funnel should be covered over writh a circular plate of glass, and the pro- cess of filtration must be repeated, if necessary, until a pale, straw- coloured, perfectly transparent filtrate results.- The sterilised test-tubes are filled to about a third of their depth by pouring in the gelatine carefully and steadily, or by employing a small sterilised glass funnel. The object of this care is to prevent the mixture touching the part of the tube with which the plug comes into contact ; otherwise, when the gelatine sets, the cotton- wool adheres to the tube and becomes a source of embarrassment in subsequent procedures. As the tubes are filled they are placed in the test-tube basket, and must then be sterilised. They are either lowered into the steam steriliser, when the thermometer indicates 100° 0., for twelve minutes for four or five successive days, or they FIG. 31. — HOT-WATER FILTERING APPARATUS. NUTRIENT MEDIA AND METHODS OF CULTIVATION. 103 may be transferred to the test-tube water-bath, and heated for an hour a day for three successive days. If the gelatine shows any turbidity after these processes it must FIG. 32.— METHOD OF MAKING A FOLDED FILTER. be poured back from the test-tubes into a flask, boiled up for ten minutes, and filtered once more, and the processes of sterilisation just described must be repeated. FIG. 33. — STEAM STERILISED Nutrient Agar-agar. — Agar-agar is a substance prepared from seaweed which grows on the coasts of Japan and India, and is supplied in long crinkled strips. It boils at 90° C., and 104 BACTERIOLOGY. remains solid up to a temperature of about 45° C. It is there- fore substituted for gelatine in the preparation of a jelly for the cultivation of those bacteria which will only grow, or grow best, in the incubator at the temperature of the blood. It may also be employed at ordinary temperatures for bacteria which liquefy gelatine. The preparation is conducted on much the same principles as those already described. Instead, however, of 100 grammes of gelatine, only about 20 grammes of agar-agar are employed (1'5 to 2 per cent.), and to facilitate its solution it must be allowed to soak in salt water overnight. For the filtration, flannel is substituted for filter- paper, or may be used in combination with the latter. The hot-water apparatus is invariably employed, unless, to accelerate the process, the glass funnel and receiver are bodily transferred to the steam steriliser. If the conical cap cannot be replaced, cloths laid over the mouth of the steriliser must be em- ployed instead. It may be necessary to repeat the process of filtra- tion, but it must not be expected that such a brilliant transparency can be obtained as with gelatine. The final result, when solid, should be colourless and clear; but if slightly milky, it may still be employed. A little liquid gradually collects in the tubes, being expressed by the contraction of the agar-agar. Wort-gelatine is used in studying the bacteria of fermentation. It is made by adding from 5 to 10 per cent, of gelatine to beer- wort. Glycerine Agar-agar. — This is prepared by adding 5 per cent, of glycerine to nutrient agar-agar, after the boiling and before the filtration, and other modifications can be made for special purposes by the addition of grape-sugar or of gelatine. FIG. 34. — INCUHATOR. NUTRIENT MEDIA AND METHODS OF CULTIVATION. 105 After the final treatment in the steam steriliser some of the of gelatine and agar-agar are placed upright and allowed to set, and others are placed on an inclined plane or in the blood-serum in.Npissator, and left to gelatinise with an oblique surface. (B) METHODS OF EMPLOYING NUTRIENT JELLY IN TEST-TUBES AND ON GLASS PLATES. Test-tube-cultivations. — To inoculate test-tubes containing nutrient jelly, the cotton- wool plug is first twisted round in case there are any adhesions between the plug and the test-tube. It is then removed with the thumb and index finger of the right hand, and placed between the fourth and fifth fingers of the left hand, instead of being put down on the laboratory table and thereby probably contaminated with bacteria or the spores of mould fungi. A sterilised needle charged, for example, with blood or pus con- taining bacteria, or with a colony from a plate-culture, is thrust once in the middle line into the nutrient jelly, and steadily withdrawn. The tube should be held horizontally or with • its mouth downward, to avoid, as far as possible, accidental contamination from the gravitation of germs in the j , , . i i FIG. 35. — METHOD OF IXOCULAT- air; and the plug replaced as quickly ^ A TJMJT.TCBB COXTAINING as possible. The cotton- wool project- STERILE NUTRIENT JELLY. ing beyond the mouth of the tube is then thoroughly burnt in the flame of a Bunsen burner or blow- pipe, and an india-rubber cap fitted over the mouth of the tube The chances of error arising from contamination of the culti- vations are reduced by avoiding draughts at the time of inoculation, and it is best that these manipulations should be carried on in a quiet room in which the tables and floor are wiped with damp cloths, rather than in a laboratory in which the air becomes charged with germs through constant sweeping and dusting, and the entrance and exit of classes of students. In conducting any investigation a dozen or more tubes should be inoculated, and if by chance an adventitious germ, in spite of all precautions, gains an entrance, 106 BACTERIOLOGY. the contaminated tube can be rejected, and the experiments con- tinued with the remaining pure cultivations. When, however, one tube containing a liquid medium is in- oculated from another, as in the process of preparing plate-cultures, or when a culture is made from a tube in which the growth has liquefied the gelatine, it is obvious that the tubes cannot be inverted or held horizontally, and they must then be held and inoculated as in Fig. 38. To inoculate those tubes of nutrient media which have been solidified obliquely, the point of a straight sterilised needle charged with the material to be cultivated is traced over the surface of the jelly from below upwards, or the inoculated material may be spread out with a hooked or looped needle. Examination of Test-tube-cultivations. — The appearances pro- duced by the growths in test-tubes can be in most cases sufficiently examined with the naked eye. In some cases the jelly is partially or completely liquefied, while in others it remains solid. The growths may be abundant or scanty, coloured or colourless. The nutrient jelly may itself be tinged or stained with products resulting from the growth of the organisms. When liquefaction slowly takes place in the nesdle track, or the organism grows without producing this change, the appearances which result are often very delicate, and in some cases very characteristic. The appearance of a simple white thread, of a central thread with branching lateral filaments, of a cloudiness, or of a string of beads in the track of the needle, may be given as examples. In some cases much may be learnt by examining the growth with a magnifying glass. Here, however, a difficulty may be encountered, for the cylindrical form of the tube so distorts the appearance of its contents, that the examination is rendered somewhat difficult. To obviate this, a very simple contrivance may be employed with advantage. This consists of a rectangular vessel, about four inches in height and two inches in width, which may be easily constructed by cementing together two slips of glass to form the back and front, with three slips of stout glass with ground edges forming the sides and base (Cheshire). The front may be constructed of thin glass, and the base of the vessel made to slope so that the test-tube when placed in the vessel has a tendency to be near the front. The vessel is filled with a mixture of the same refractive index as the nutrient gelatine. The latter has a refractive index rather higher than water, which is about 1'333 ; alcohol has a ref reactive index of 1'374. The vessel is filled with water, and alcohol is then added until the proper density is reached. The test-tube is placed in the NUTRIENT MEDIA AND METHODS OF CULTIVATION. 107 vessel, and held in position by means of a clip. The vessel can be fixed on the inclined stage of the microscope, and the contents of the tube conveniently examined with low-power objectives. Plate- cultivations. — By this method, as already mentioned, a mixture of bacteria, whether in fluids, excreta, or in cultivations on solid media, can be so treated that the different species are isolated one from the other, and perfectly pure cultivations of each of the cultivable bacteria in the original mixture established in various nutrient media. We are enabled also to examine under a low power of the microscope the individual colonies of bacteria, and to distinguish by their characteristic appearances, micro-organisms which, in their individual form, closely resemble one another, or are even identical. The same process, with slight modification, is also employed in the examination of air, soil, and water, which will be referred to later. The preparation of plate-cultivations, therefore, must be described in every de- tail : and to take an example, we will sup- pose that a series of plates is to be pre- pared from a test- cube- cultivation. Arrangement of Levelling Apparatus. — In order to spread out the liquid jelly evenly on the surface of a glass plate, and hasten its solidifica- tion, it is necessary to place the glass plate upon a level and cool surface. This is obtained in the following manner : Place a large shallow glass dish upon a tripod stand, and fill it to the brim with cold water ; carefully cover the dish with a slab of plate-glass, or a pane of window-glass, and level it by placing the spirit-level in the centre and adjusting the screws of the tripod. Substitute for the spirit-level a piece of filter-paper the size of the glass plates to be employed, and cover it with a shallow bell-glass. Sterilisation of Glass Plates. — The glass plates are sterilised in an iron box placed in the hot-air steriliser, at 150° C., from one to two hours. As these plates are used also for other purposes, a quantity ready sterilised should always be kept in the box. Preparation of Damp Chambers. — The damp chambers for the reception of the inoculated plates are prepared thus : Thoroughly FIG. 36. — LEVELLING APPARATUS. 108 BACTERIOLOGY. cleanse and wash out with 1 in 20 carbolic acid a shallow glass dish and bell. Cut a piece of filter-paper to line the bottom of the glass dish, and moisten it with the same solution. Method of Inoculating the Test-tubes. — In a glass beaker or an ordinary glass tumbler, with a pad of cotton-wool at the bottom, place the tube containing the cultivation, the three tubes to be inoculated, three glass rods which have been sterilised by heating in the flame of a Bunsen burner, and a thermometer. Provide a strip of paper, a large label, a pencil, a pair of forceps, and inoculating needles. All is now ready at hand to commence the inoculation of the tubes. Liquefy the gelatine in the three tubes by placing them in a beaker containing water at 30° C., or by gently warming them in the flame of the Bunsen burner. Keep the tubes, both before and after the inoculation, in the warm water, to maintain the gelatine in a state of liquefaction. Hold the tube containing the cultivation FIG. 37. — IRON Box GLASS PLATES. FOR FIG. 38.— METHOD OF INOCULATING TEST-TUBES IN THE PREPARATION OF PLATE-CULTIVATIONS. and a tube of the liquefied gelatine as nearly horizontal as possible between the thumb and index finger of the left hand. With the index finger and thumb of the right hand loosen the plugs of the tubes. Take the looped platinum needle in the right hand and hold it like a pen. Remove the plug from the culture- tube by using the fourth and fifth fingers of the right hand as forceps, and place it between DESCRIPTION OF PLATE III. Plate-cultivation. This represents the appearance of a plate-cultivation of the comma-bacillus of Cholera nostras, when it is examined over a slab of blackened plate-glass. The drawing was made from a typical result of thinning out the colonies by the process of plate-cultivation. At this stage they were completely isolated one from the other ; but later they became confluent, and produced complete liquefaction of the gelatine. Plate IE. fc o f-H H t i— i H H P O NUTRIENT MEDIA AND METHODS OF CULTIVATION. 109 the fourth and fifth fingers of the left. Remove the plug of the other tube in the same way, placing it between the third and fourth fingers of the left hand. With the needle take up a droplet of the cultivation and stir it round in the liquefied jelly. Replace the plugs, and set aside the cultivation. Hold the freshly inoculated tube be- tween the index finger and thumb of either hand, almost horizontally, then raise it to the vertical, so that the liquid gelatine gently flows back. By repeating this motion and rolling the tube between the fingers and thumbs the micro-organisms which have been introduced are distributed throughout the gelatine. Any violent shaking, and consequent formation of bubbles, must be carefully avoided. From the inoculated tube, in the same manner inoculate a fresh tube of liquefied gelatine, introducing into it three droplets with a sterilised needle. After tilting and rolling this tube, as in the previous case, the same process is repeated with a third tube, which is inoculated from the second tube. This last tube must be inoculated in different ways, according to experience, for different micro-organisms. Some- times a sufficient separation of the micro-organisms is attained by inoculating the last tube with a straight, instead of a looped, needle, dipping it from the one into the other from three to five times. The next process consists in pouring out the gelatine on glass plates and allowing it to solidify. Preparation of the Gelatine- Plates. — The directions to be observed in pouring out the gelatine are as follows : — Place the box containing sterilised plates horizontally, and so that the cover projects beyond the edge of the table ; remove the cover, and withdraw a plate with sterilised forceps ; hold it between the finger and thumb by opposite margins, rapidly transfer it to the filter-paper under the bell-glass, and quickly replace the cover of the box. On removing the plug from the tube which was first inoculated, an assistant raises the bell-glass, and the contents of the tube are poured on to the plate ; with a glass rod the gelatine must be then rapidly spread out in an even layer within about half an inch of the margin of the plate. The assistant replaces the bell- glass, and the gelatine is left to set. Meanwhile a glass bench or metallic shelf is placed in the damp chamber, ready for the reception of the plate-cultivation, and when the gelatine is quite solid the plate is quickly transferred from under the bell-glass to the damp chamber ; precisely the same process is repeated with the other tubes, and the damp chamber, labelled with the details of the experiment, is set aside for the colonies to develop. Not only plate- cultures should be carefully labelled with date and description, but 110 BACTERIOLOGY. the same remark applies equally to all preparations — tube- cultures, potato-cultures, drop-cultures, etc. Corresponding with the fractional cultivation of the micro- organisms obtained in this manner, the colonies will be found to develop in the course of a day or two, the time varying with the temperature of the room. The lower plate will contain a countless number of colonies which, if the micro-organism liquefies gelatine, speedily commingle, and produce, in a very short time, a complete liquefaction of the whole of the gelatine. On the middle plate the colonies will also be very numerous, but retain their isolated position for a longer time ; while on the uppermost plate the colonies are completely isolated from one another, with an appreciable surface of gelatine intervening. Examination of Plate- cultivations. — The macroscopical appear- ances of the colonies are best studied by placing the plate on a FIG. 39. — DAMP CHAMBER CONTAINING PLATE-CULTIVATIONS. slab of blackened glass, or on a porcelain slab if the colonies are coloured. To examine the microscopical appearances, a selected plate is placed upon the stage of the microscope. The smallest diaphragm is employed, and the appearances studied principally with a low power. These appearances should be carefully noted, and a sketch or photograph of the colony made. The morphological characteristics of the micro-organisms of which the colony is formed can be examined in the following way : A small looped or hooked platinum needle is held like a pen. and the hand steadied by resting the little finger on the stage of the microscope. The extremity of the needle is steadily directed to the space between the lens and the gelatine without touching the latter, until, 011 looking through the microscope, it can be seen in the field, above or by the side of the colony under examination. The needle is then dipped into the colony, steadily raised, and withdrawn. Without removing the eye from the microscope this manipulation can be seen to be successful by the colony being disorganised or NUTRIENT MEDIA AND METHODS OF CULTIVATION. Ill completely removed from the gelatine. It is, however, not easy to lu« successful at first, but with practice this can be accomplished with rapidity and precision. A preparation is then made by rubbing FIG. 40.— PASTEUR'S LARGE INCUBATOR. the extremity of the needle in a droplet of water on a slide, covering with a cover-glass, and examining in the fresh state, or by spreading out the droplet on a cover-glass, drying, passing three times through the flame, and staining with a drop of fuchsine or gentian violet. Inoculations should be made in test-tubes of nutrient gelatine OF THF. UNIVERSITY 112 BACTERIOLOGY. and agar-agar, from the micro-organisms transferred to the cover- glass before it is dried and stained, from any remnants of the colony which was examined, or from other colonies bearing exactly similar appearances. In this way pure cultivations are established, and the rnacroscopical appearances of the growth in test-tubes can be obtained. The plates should be replaced in the damp chamber as soon as possible ; drying of the gelatine, or contamination with micro-organisms gravitating from the air during their exposure, may spoil them for subsequent examination. A much simpler method of plate-cultivation is to dispense with the levelling apparatus, and pour the liquefied ielly into shallow, flat dishes. FIG. 41.— PETRI'S J J j)ISH< -They take up much less room, and in many ways are more convenient (Fig. 41). Nutrient agar-agar can also be employed for the preparation of plate-cultivations, but it is much more difficult to obtain satis- factory results. The test-tubes of nutrient agar-agar must be placed in a beaker with water and heated until the agar-agar is completely liquefied. The gas is then turned down, and the temperature of the water allowed to fall until the thermometer stands just above 50° C. The water must be maintained at this temperature, and the test-tubes must be in turn rapidly inoculated and poured out upon the glass plates, or better still, into glass dishes, as already described. A very much simpler plan is to liquefy the agar, pour it into FIG. 42. — GLASS BENCHES AND SLIDES. a shallow dish, and allow it to solidify. The culture material is thinned out in sterilised broth, and a few drops are spread out over the surface of the agar. The dishes are then placed in the incubator at 37° C. Glass plates may also be employed in a much simpler way. The nutrient jelly is liquefied, poured out, and allowed to set. A needle charged with the material to be inoculated is then drawn in lines over the surface of the jelly. This method is of use for inoculating different organisms side by side, and watching the effect of one upon the other, or a micro-organism in this way may be NUTRIENT MEDIA AND METHODS OF CULTIVATION. 1 13 sown upon the gelatine which has been already altered by the growth of another micro-organism ; the change produced in the gelatine, as in the case of the Bacillus pyocyaneus, extending far beyond the limits of the growth itself. Nutrient jelly may also be spread out on sterilised glass slides, which after inoculation are placed in damp chambers for the growths to develop. Esmareh's Roll-cultures. — Esmarch introduced a modification of the method of plate- cultivation which may sometimes be used with advantage. The ordinary test-tubes may be employed, or tubes considerably larger in size. After the liquid jelly has been inoculated in the tube, instead of pouring it out on to a glass plate or into a dish, the cotton-wool plug is replaced, and an india-rubber cap fitted over the mouth of the tube. The tube is then placed horizontally on a block of ice or in a vessel containing iced water. The neck of the tube is steadied with the left hand, and the tube turned round and round with the right hand. In a very short time the gelatine sets, and the tube is lined inside with a thin coating. There is far less danger of contamination, and the cultures are in a much more convenient form when circumstances render it necessary to move them. (c) PREPARATION AND EMPLOYMENT OF SOLIDIFIED BLOOD SERUM. Solid Blood Serum.— The tubercle-bacillus, the bacilli of glanders and of diphtheria, and many other micro-organisms, thrive well when cultivated on solid blood serum. This medium has the additional advantage of remaining solid at all temperatures. The technique required for its preparation and sterilisation is as follows : Several cylindrical vessels, about 20 cm. high, are thoroughly washed with carbolic acid (1 in 20), and then with alcohol, and finally rinsed out with ether. The ether is allowed to evaporate, and the vessels are then ready for use. The skin of the animal selected — calf, sheep, or horse — is washed with carbolic at the seat of operation, and the bleeding performed with a sterilised knife or a trocar and canuula. The first jet of blood from the vein is rejected, and that which follows is allowed to flow into the vessels until they are almost full. The ground-glass stoppers, greased with vaseline, are replaced, and the vessels set aside in ice, as quickly as possible, for from twenty-four to thirty hours. By that time the separation of the clot is completed, and the clear serum can then be transferred to 8 114 BACTERIOLOGY. plugged sterilised test-tubes. These should be filled, with a sterilised pipette, to about one-third of their capacity. Formerly the tubes were sterilised by Tyndall's process of dis- continuous sterilisation. The tubes were placed in Koch's serum steriliser, with the temperature maintained for an hour or more at 56° C., and this was repeated for six successive days, the temperature on the last day being gradually raised to 60° C. This completed the sterilisation, and to solidify the serum the tubes were arranged in the inspissator at the angle required, and the temperature was kept between 65° C. and 68° C. Directly solidification took place the tubes were removed. The new process is much less tedious, and consists in taking every possible pre- caution to obtain the blood without contamination by bac- teria in the air or in the vessels employed. There is then no need to sterilise the serum, and it can be coagu- lated immediately. The tubes are tested by placing them in an incubator at 37° C. for a week, and if any show signs of contamination they are discarded, and the rest can be used or kept in stock. The serum should then present the character of being hard, solid, of a pale straw colour, and transparent. A little liquid collects at the lowest point, and the serum is sometimes milky in appearance at -its thickest part. Loffler's Blood Serum is prepared by mixing two- thirds of fresh serum with one-third of broth, prepared in the usual way but with the addition of 1 per cent, grape-sugar. The mixture is decanted into test-tubes, avoiding the formation of air- bubbles, and it is then coagulated in the usual way. The serum may be employed not only in test-tubes, but also in small flasks, glass capsules, or other vessels, all of which must be cleansed and sterilised. Hydrocele fluid and other serous effusions may be prepared in the same manner. (Gelatine may be added to the serum in the pro-- portion of 5 per cent. FIG. 43. — KOCH'S SERUM STERILISER. NUTRIENT MEDIA AND METHODS OF CULTIVATION. 115 Inoculation of the Tubes. — A small portion of a culture or of the material to be inoculated is taken up with a sterilised platinum needle, and traced over the sloping surface of the serum ; or a fragment of tissue, such as diphtheritic membrane or tubercle, may be introduced into the tube and rubbed gently over the serum so as not to break the surface. FIG. 44. — HUEPPE'S SERUM INSPISSATOR. (D) PREPARATION AND EMPLOYMENT OF STERILISED POTATO. Potato-cultivations. — Sterilised potatoes form an excellent medium for the cultivation of many micro-organisms, more especially the chromogenic species. Potato -cultivations also give in some cases very characteristic appearances, which are of value in distinguishing bacteria which possess morphological resemblances. Preparation of Sterilised Potatoes. — Potatoes, preferably smooth - skinned, which are free from "eyes" and rotten spots, should be selected. If they cannot be obtained without eyes and spots, these must be carefully picked out with the point of a knife. The potatoes are well scrubbed with a stiff brush, and allowed to soak in 1 in 20 carbolic for a few minutes. They are then transferred to the potato- 116 BACTERIOLOGY . receiver, and steamed in the steam steriliser for twenty minutes to half an hour, the time varying according to the size of the potatoes. When cooked, the potato-receiver is withdrawn and left to cool, the potatoes being retained in it until required for use. Damp chambers are prepared ready for the potatoes, the vessels being cleansed and washed with carbolic as described for plate- cultivations. Small glass dishes of the same pattern as the large ones may be employed for single halves of potatoes. Potato-knives and scalpels, which have been sterilised in an iron box by heating them in the hot-air steriliser at 151° C. for one hour, should be ready to hand. Knives sterilised by heating them in the name of a Bunsen burner should afterwards be placed upon a sterilised glass plate and covered with a bell-glass. It must not be forgotten, FIG. 45. — Box FOR STERILISING INSTRUMENTS. however, that heating the blades in the flame destroys the temper of the steel, and therefore knives and other instruments should preferably be sterilised in the hot-air steriliser, enclosed in an iron box, or simply enveloped in cotton-wool. Inoculation of Potatoes. — The coat-sleeves should be turned back, and the hands, after thorough washing with good lathering soap, be dipped in 1 in 40 carbolic. An assistant opens the potato- receiver, and a potato is selected and held between the thumb and index linger of the left hand. With the knife held in the right hand, the potato is almost completely divided in the direction which will give the largest surface. The assistant raises the cover of the damp chamber, and the potato is introduced, and while the knife is withdrawn, allowed to fall apart. The cover is quickly replaced, and another potato treated in the same way, is placed in the same damp chamber. The four halves are then quite ready for inoculation. As an extra precaution, the left hand is again dipped in carbolic, and one half of a potato is taken up between the tips of the thumb and index finger, care being taken to avoid touching the cut surface. Holding it with its cut surface vertical, NUTRIENT MEDIA AND METHODS OF CULTIVATION. 117 a small portion of the substance to be inoculated is placed on the centre with a sterilised platinum needle. With a sterilised scalpel the inoculated substance is rapidly spread over the surface of the potato with the flat of the blade, to within a quarter of an inch of the margin, and the potato is then as quickly as possible replaced in the damp chamber. With another sterilised scalpel a small portion of the potato from the inoculated surface of the first half is in the same way spread over the surface of the second half, thus thinning out the bacteria as in plate-cultivations. Exactly the same is repeated with a third potato, and even a fourth, so that a still further thinning out or fractional cultivation of the micro- organisms may be obtained. In some cases it is necessary to place the cultures in an incubator (Fig. 40) ; others grow very well at the FIG. 46. — DAMP CHAMBER FOR POTATO-CULTIVATIONS. temperature of the room. As in plate-cultivations, the potato may also be inoculated by simply streaking it in lines with a needle charged with the material to be cultivated. Potato in Test-tubes. — Large surfaces of potato are employed when we wish to obtain cultures of micro-organisms in considerable quanti- ties, as in the examination of the products of chromogenic bacteria ; but under ordinary circumstances potato is employed in test-tubes. The central portions of raw potatoes are cut out in cylindrical pieces with a cork-borer. These are divided obliquely in their whole length, and each half is placed in a test-tube. The test-tubes are plugged with cotton-wool, and then steam in the steam-steriliser for twenty minutes. The sloping surface is inoculated in the same way as obliquely solidified jelly, and the advantages are great. The cultures are obtained in a more convenient form, and there is less danger of contamination. Potato-paste may be employed when it is desirable to obtain an extensive growth of certain bacteria. The potatoes are boiled for an hour, and the floury centre squeezed out of the skins. This is then mashed up with sufficient sterilised water to produce a thick 118 BACTERIOLOGY. paste, and is heated in the steam steriliser for half an hour for three successive clays. (E) PREPARATION AND EMPLOYMENT OF BREAD-PASTE, VEGETABLES, FRUIT, WHITE OF EGG. Some micro-organisms, more especially mould fungi, grow very well on bread-paste. This is prepared by removing the crust from slices of bread and drying them in the oven. They are then broken up, and reduced to a fine powder with a pestle and rnortar. Small, carefully cleansed, conical, or globe-shaped flasks are plugged with cotton-wool and sterilised in the oven. When cool, a small quantity of the powder is placed in them, and sterilised water added in the proportion of one part to every four of the powder. The paste is sterilised by steaming in the steriliser at 100° C. for half an hour for three successive days. The flasks can be reversed, and may be inoculated with a platinum needle. Boiled carrots and other vegetables, and various kinds of stewed fruit, are also occasionally employed for the cultivation of bacteria. The sterilisation of these media must be carried out on the principles already explained. White of egg may be solidified in shallow glass dishes, in the steam steriliser. After inoculation the dishes should be placed in a damp chamber. LIQUID MEDIA. (F) PREPARATION OF STERILISED BROTH, LIQUID BLOOD SERUM, URINE, MILK, VEGETABLE INFUSIONS, AND ARTIFICIAL NOURISH- ING LIQUIDS. Nutrient liquids are still largely employed. For inoculation ex- periments when the presence of gelatine is undesirable, for studying the physiology and chemistry of bacteria, and when for any object a rapid growth of micro-organisms is necessary, the employment of liquid media is not only advisable, but absolutely necessary. Liquid media comprise two distinct groups — natural and artificial. Natural media include meat broth, blood, urine, milk, and vegetable infusions ; artificial media are solutions composed from a chemical formula representing essential food constituents. Broth may be made from beef, pork, chicken, or fish in the manner which has been described for the preparation of nutrient gelatine, simply with omission of the gelatine. After the process NUTRIENT MEDIA AND METHODS OF CULTIVATION. 119 of neutralisation with carbonate of soda solution, the flask of broth is placed in\ the steam steriliser for half an hour at 100° C. A clear liquid results on nitration which is transferred to plugged sterilised flasks or test-tubes, and sterilisation effected by exposing the*n in the steam steriliser for half an hour at 100° C. for two FIG. 47. — APPARATUS FOE STERILISATION BY STEAM UNDER PRESSURE. or three successive days, or] by; I using the apparatus for sterilising by steam under pressure. For some bacteria a more suitable cultivating medium is obtained by the addition of glycerine or grape-sugar. Liquid Blood Serum. — The preparation of blood serum has already been described. ' It may be required for cultivations before the final treatment by which it is solidified, for example, in the method of drop-cultivation, and may be used with the addition of glycerine or grape-sugar. Hydrocele fluid, peritonitic arid pleuritic 120 BACTERIOLOGY. effusions, can also be employed after sterilisation in the steam steriliser. The fluid should be withdrawn with a sterilised trocar and cannula, and received into plugged sterilised flasks. Urine. — -In order to obtain urine free from micro-organisms the following precautions must be observed : The orifice of the urethra must be thoroughly cleansed with weak carbolic. The first jet of urine should be rejected, and the rest received into sterilised vessels, which must be quickly closed with sterilised plugs. If these pre- cautions be not attended to, the urine must be rendered sterile by the means described for the sterilisation of broth. Milk. — If milk has been drawn into sterile flasks, after thoroughly cleansing and disinfecting the teats and hands, it may be kept without change. If procured without these precautions, it must be steamed in the steriliser for half an hour for five successive days. Vegetable and other Infusions. — Infusions of hay, cucumber, and turnip are used for special purposes, and more rarely decoctions of plums, raisins, malt, and horse-dung. They are mostly prepared by boiling with distilled water, after maceration for several hours. The filtrate is received into sterile flasks and sterilised in the usual way in the steam steriliser. Artificial Fluids. — Pasteur's solution is prepared by mixing the ingredients in the following proportions : — Distilled water .100 Pure cane-sugar . . . . . . 10 Ammonium tartrate ..... 1 Ash of yeast '075 Mayer's modification of the nourishing fluid employed by Colin is as follows : — Distilled water 20 Ammonium tartrate "2 Phosphate of potassium . . . . "1 Sulphate of magnesium ... "1 Tribasic calcium phosphate .... '01 Drop- cultures. — This method of cultivation is a particularly instructive one. It enables us to study many of the changes which take, place during the life history of micro-organisms. This is illustrated, for example, in a drop-culture of the anthrax bacillus, in which we can watch the gradual growth of a single bacillus into NUTRIENT MEDIA AND METHODS OF CULTIVATION. 121 a long filament, and the subsequent development of bright oval spores. It is necessary carefully to observe the minutest details in order to maintain the cultivation pure. An excavated slide is thoroughly cleaned, and then sterilised by being held with the cupped side downwards in the flame of the Bunsen burner. A ring of vaseline is painted round the excavation, and the slide is then placed under a glass bell. Meanwhile a carefully cleansed cover- glass is also sterilised by passing it through the flame, and should be deposited on a sterilised glass plate. With a sterilised looped needle, a drop of sterile broth is transferred to the cover-glass, and this is inoculated by touching it with another sterilised needle charged with the material to be examined, without disturbing the form of the drop. It is quite sufficient just to touch the drop FIG. 48. — DROP CULTIVATION. (a) Drop of broth ; (6) layer of vaseline. instead of transferring a visible quantity of blood, juice, or growth, as the case may be. The slide is then inverted and placed over the cover-glass, so that the drop will come exactly in the centre of the excavation, and is gently pressed down. On turning the slide over again the cover-glass adheres, and an additional layer of vaseline is painted round the edges of the cover-glass itself. The slide must be labelled, and if necessary, placed in the incubator, and the results watched from time to time. Instead of broth, liquid blood serum may be employed in this form of cultivation. If it is required to preserve the drop-cultivation as a microscopic preparation, the cover-glass is gently lifted off and allowed to dry. Any vaseline adhering to the cover-glass should be wiped off, and the cover-glass can then be passed through the flame and stained in the usual manner. Moist Cells. — Unless drop-cultures are very carefully prepared 122 BACTERIOLOGY. they are liable to dry up, if kept for examination for several days. Many therefore prefer employing a moist cell, of which there are several different forms in use. The drop-culture slide may be converted into a moist cell by having a deep groove cut round the circumference of the con- cavity. This groove is filled with sterilised water by means of a pipette. A ring of vaseline is painted with the camel's-hair brush outside the groove, and the cover-glass, with the drop -cultivation, is inverted and placed over the concavity. This form is very useful, as the slide can be easily cleansed and effectually sterilised by holding it in the flame of the Bunsen burner. A very simple form of moist cell recommended by Schafer FIG. 49. — SIMPLE METHOD OF FOKMING A MOIST CELL. may be used in some cases, but possesses the disadvantage of not admitting of sterilisation by heat. A small piece of putty or modelling wax is rolled into a cord about two inches long and J inch thick. By uniting the ends a ring is formed, which is placed on the middle of a clean glass slide. A drop of water is placed in the centre of the ring, and the cell roofed in by applying a cover-glass. A cell somewhat similar in form, which has the advantage of permitting of thorough cleansing, may be constructed by cementing a glass ring with flat surfaces to an ordinary slide. Vaseline is applied with a camel's-hair brush to the upper surface of the ring, and one or two drops of water placed with a pipette at the bottom of the cell. The cover-glass, with the preparation, is then inverted over the cell and gently pressed down upon the glass ring. The vaseline renders the cell air-tight, and, to a certain extent, fixes the cover-glass to the ring. NUTRIENT MEDIA AND METHODS OF CULTIVATION. 123 Warm Stages. — To apply warmth while a preparation is under continuous observation, we must either place the microscope bodily FIG. 50.— WARM STAGE. within a special incubator, with the eye-piece protruding through an opening, or we must employ some means of applying heat directly to the preparation. A simple warm stage may be made of an oblong copper plate, FIG. 51. — WARM STAGE SHOWN IN OPERATION. two inches long by one inch wide, from one side of which a rod of the same material projects. The plate has a round aperture in the 124 BACTERIOLOGY. middle, half an inch in diameter, and is fastened to an ordinary slide with sealing-wax. The drop to be examined is placed on a large-sized cover-glass and covered with a smaller one. Olive oil or vaseline is painted round the edge of the smaller cover-glass to prevent evaporation, and the preparation is placed over the aperture in the plate. The slide bearing the copper plate is clamped to the stage of FIG. 52.— ISRAEL'S WARMING APPARATUS IN OPERATION. the microscope. The flame of a spirit-lamp is applied to the extremity of the rod, and the heat is conducted to the plate and thence transmitted to the specimen. In order that the temperature of the copper plate may be approximately that of the body, the lamp is so adjusted that a fragment of cacao butter and wax, placed close to the preparation, is melted. Israel's Warming Apparatus. — It is obvious that in employing very high powers a difficulty will be presented by the warm stages NUTRIENT MEDIA AND METHODS OF CULTIVATION. 125 commonly used for accurate observations, such as Schafer's or Strieker's, owing to their interference with the illumination. To FIG. 53.— SECTION OF ISRAEL'S WARMING APPARATUS AND DROP-CULTURE SLIDE, overcome this an apparatus has been constructed by which the slide is warmed from above (Figs. 52, 53). The drop-culture slides are provided with a shallow groove, *1 mm. deep and 1 mm. broad, cut round the concavity. Into this the cover-glass fits, so that its upper surface is level with that of FIG. 54. — ISRAEL'S WARMING APPARATUS. the slide. The heating apparatus consists of a flat disk-shaped box with a central conical aperture. The entrance and exit pipes are fixed on at a right angle to the 126 BACTERIOLOGY. side (Fig. 54). The former, z, is of metal, ancLthe latter, a, of glass fitted with a thermometer, the bulb of which, k, is contained within the box. A partition, s, keeps up a current between the openings FIG. 55. — GAS CHAMBER IN USE WITH APPARATUS FOR GENERATING CARBONIC ACID. of the pipes, which are supported on a stand and connected by tubing with the hot-water supply. A mixture of paraffine and vaseline is recommended for indicating the temperature of the chamber, and experience has shown that if a temperature of 37° C. is required the temperature of the water in the box must range between 42° and 47° C. FIG. 56.— GAS CHAMBER. Gas Chambers. — To investigate the action of gases or vapours upon micro-organisms, a modification of the moist cell may be employed. A piece of glass tubing is first fixed to the slide by means of NUTRIENT MEDIA AND METHODS OF CULTIVATION. 127 sealing-wax, and the ring of putty is so placed as to include one end of it, leaving a small interval at the side, or a little notch is made in the putty opposite, so as to afford an exit for the gas or vapour. FIG. 57.— MOIST CELL ADAPTED FOR TRANSMISSION OF ELECTRICITY. Application of Electricity. — To study the effect of electricity we may prepare a drop-culture in the moist cell. The cover- glass to be used is provided with two strips of tinfoil, which are FIG. 58.— APPARATUS ARRANGED FOR TRANSMITTING ELECTRICITY. isolated from the brass of the microscope, and so arranged that a current of electricity may be passed through them. A much simpler plan, which may also be employed, is to take an ordinary glass slide and coat the surface with gold-size. The 128 BACTERIOLOGY. slide is then pressed firmly down on gold-leaf or tinfoil and allowed to dry. When dry, the metal is scraped away, leaving two triangles with a small interval between them. FIG. 59. — SLIDE WITH GOLD-LEAF ELECTRODES. The liquid containing the micro-organisms is placed between the electrodes, covered with a cover-glass, and then subjected to the electric current. (G) METHODS OF EMPLOYING AND STORING LIQUID MEDIA. Cultivations in liquid media can be carried on in test-tubes, but it is more satisfactory to employ special forms of flasks, bulbs, and U tubes, such as those employed by Pasteur and his school, and by Lister, Sternberg, and Aitken. Lister's flasks. — These flasks were especially introduced by Lister as a means of storing liquid nutrient media. They are so constructed that after removal of a portion of the contents, on restoring the vessel to the vertical position, a drop of liquid always remains in the extremity of the nozzle, which prevents regurgitation of unfiltered air, Sternberg' s Bulbs. — The method of intro- ducing liquid into the. bulb employed by Sternberg, and of sterilising and inoculating it, is as follows : The bulb is heated slightly over the flame, and the extremity of the neck, after the sealed point has been broken off, is plunged beneath the surface of the liquid. As the air cools the liquid is drawn into the bulb, usually filling it to about one-third of its capacity. C^. .— - — The neck of the flask is again sealed up, and the liquid which has been introduced is sterilised by FlG' G|--STERX- BERG s BULB. repeatedly boiling the flasks in the water- bath. They should then be placed in the incubator for two or three days ; FIG. 60.— LISTER'S FLASK. NUTRIENT MEDIA AND METHODS OF CULTIVATION. 129 Fi 62.— AITKEN'S TUBE. i\nd if the contents remain transparent and free from film, they may be set aside as stock-bulbs, to be used when required. To inoculate the liquid in the bulb the end of the neck is heated to sterilise the exterior, the bulb is gently warmed, and the extremity of the neck nipped off with a pair of sterilised forceps. The open extremity is plunged into the liquid containing the micro-organisms, and a minute quantity enters the tube and mingles with the fluid in the bulb without fear of contamination by atmospheric germs. The extremity of the neck is once more sealed up in the flame of a Bunsen burner. Aitkeris Tubes. — These tubes are plugged and sterilised, and the nutrient medium introduced as into ordinary test-tubes. Instead of withdrawing the cotton-wool plug, they are inoculated through a lateral arm. The sealed extremity of the arm is nipped off with sterilised forceps, and the inoculating needle is carefully introduced through the opening thus made. It is directed along the arm until it touches the opposite side of the test-tube, where it deposits the material with which it was charged. The needle is withdrawn, and the end of the lateral arm again sealed up in the flame ; the test-tube is then tilted until the liquid touches the deposited material ; on restoring the tube to the vertical, the material is washed down with the nutrient liquid. Miguel's Bulbs. — The tube a boule of Miquel is also a very useful form. It consists of a bulb of 50 cc. capacity, blown in the middle of a glass tube. The part of the tube above the bulb is contracted in two places, and can either be left quite straight or made to curve slightly. Between the contrac- tions the tube is plugged with asbestos. The portion of the tube below the bulb is $ shaped, and drawn out at its extremity into a fine ' point. The bulb is charged with nutrient liquid and inoculated by aspiration, and the point of the $ tube sealed in the flame of a Bunsen burner. 9 FIG. 63.— MIQUEL'S BULB. 130 BACTERIOLOGY. FIG. 64. — PASTEUR'S FLASK. Pasteur's Apparatus. — Special forms of tubes, bulbs, and pipettes are employed by the school of Pasteur. The tubes are provided with lateral or with curved arms drawn out to a fine point, and with slender necks plugged with cotton-wool. A double form (Fig. 65) shaped like a tuning- fork, each limb with a bent arm, is con- venient for storing sterilised broth. The sealed end of an arm is nipped off with sterilised forceps, the sterile broth aspirated into each limb, and the arm again sealed in the flame ; a series of such tubes can be arranged upon a rack on the working table. Bulbs with a vertical neck drawn out to a fine point, others with a neck bent at an obtuse angle, plugged with cotton- wool, and a lateral curved arm drawn out to a fine point, are also employed. For a description of these various vessels and their special advantages, the works of Pasteur and Duclaux must be con- , . , FIG. 65.— PASTEUR'S DOUBLE suited. TUM, (H) CULTIVATION OF ANAEROBIC BACTERIA. To cultivate anaerobic organisms the same media are employed as for aerobic organisms, but the methods must be modified, or special apparatus used, so that the oxygen in the air may be excluded. In the preparation of plate-cultivations, before the film of gelatine has completely hardened it is covered with a sheet of mica, arid the edges are sealed with melted paraffine. By this process the air is not completely excluded, so that only those organisms which are not strictly anaerobic can be grown by this method. Liborius recom- mends boiling a considerable volume of gelatine in a tube, cooling it,. and after thoroughly distributing the organisms in the still liquid jelly, rapidly solidifying it by placing the tube in iced water. By this NUTRIENT MEDIA AND METHODS OF CULTIVATION. 131 process very little air re-enters the jelly, and colonies of even strictly anaerobic bacteria will develop in the lower part of the tube. The drawback is the difficulty encountered in examining the colonies, and in preparing sub-cultures. For this purpose the tube must be broken, or carefully warmed until the jelly can be shaken out. Esmarch first prepares a roll culture, and when the gelatine film has set, the tube is completely filled with liquefied gelatine which has been cooled down almost to the temperature at which it solidifies. The same difficulty arises as in the previous method, in the examination of the colonies. Buchner places the culture tube inside a much larger tube containing a small quantity of pyro- gallic acid and closed with a gutta-percha cap. The pyrogallic acid absorbs the oxygen, but the method is not altogether success- ful. The most satisfactory plan is to exhaust the air with an air pump, or to substitute an atmosphere of hydrogen which does not affect the growth of the bacteria. Various forms of flasks and tubes for cultivating bacteria have been devised, which can be easily con- nected with an exhausting 66>_FRA-NKEL,S ANAEROBIC TUBE-CULTURE' Glass tube through which hydrogen is passed; b, exit tube; c, india-rubber stopper coated externally with paraffin (FRANKLAND). apparatus, and readily sealed by the flame of the blowpipe when the air has been removed. If hydrogen is employed the most convenient plan is to use a Kipp's apparatus, from which the hydrogen is passed through two bottles, one containing a solution of lead, to remove any sulphuretted hydrogen, and the other pyrogallic acid, to intercept any oxygen. 132 BACTERIOLOGY. In the method recommended by Frankel a tube of gelatine is liquefied, and inoculated. A gutta-percha stopper is substituted for the cotton-wool plug (Fig. 66). It is perforated by two holes, through which two tubes pass which are bent at a right angle. One tube only just passes through the stopper, the other reaches down to the bottom of the test-tube. The horizontal part of each tube has a narrow neck. The long tube has a plug of steril- ised cotton-wool, and is connected, with a short piece of india-rubber tubing by which it can be connected with Kipp's apparatus. The hydrogen drives the air out of the liquefied jelly and out of the test-tube, and after about half an hour the horizontal tubes are sealed up, and the test-tube is made into a roll culture. Liborius employs a tube with a narrow neck and a lateral arm (Fig. 67). The tube is filled up to the height of the arm with either nutrient agar or a mixture of nutrient agar with 2 per cent, of grape- sugar. The liquefied jelly is inoculated in the usual way, and hydrogen passed through the lateral arm. When the air has been completely driven out, the tube is sealed up. To cultivate anaerobic organisms in broth, such as the tetanus bacillus, a flask is inoculated with the bacillus, and a stream of hydrogen is passed through the broth by means of a tube passing down to the bottom of the flask. The air in the flask escapes by a lateral arm which is bent down at a right angle, and immersed in a capsule of mercury. When the air has been completely expelled the entrance tube is hermetically sealed, and the mercury in the capsule prevents any air from re-entering the flask by the lateral arm (Fig. 68). FIG. 67.— ANAEROBIC CUL- TURE-TUBE (LlBORIUS). METHOD OF FIXING CULTURES. The colonies in plate -cultivations and the growths of bacteria in test-tubes may be stopped at any stage of their growth, and permanently fixed by exposing the culture to the fumes of formic NUTRIENT MEDIA AND METHODS OF CULTIVATION. 133 aldehyde. The test-tubes, dishes, or 'capsules are placed in a cylin- drical glass vessel containing a pledget of cotton-wool moistened with FIG. 68. — APPAKATUS FOR ANAEROBIC CULTURES. (ROSCOE AND LUNT.) formic aldehyde. The vessel is fitted with a ground glass stopper and set aside. The growth almost immediately ceases. Any liquefied gelatine is hardened, so that the exact appearances of cultures are obtained in a permanent form. CHAPTER X. EXPERIMENTS UPON THE LIVING ANIMAL. To carry out the last of Koch's postulates, and so complete the chain of evidence in favour of the causal relation of micro-organisms to disease, and to study the mode of action of a pathogenic bacterium, it is necessary to introduce into a living animal a pure cultivation of the micro-organism or its chemical products. For this purpose various animals are employed, such as mice, rabbits, guinea-pigs, pigeons, and fowls. Inhalation. — The animals may be made to inhale an atmosphere impregnated with micro-organisms by means of a spray. In this way Priedlander succeeded in administering the bacteria of pneu- monia to mice ; and the production of tuberculosis by experimental inhalation has thrown light upon the clinical records of cases reported as instances o£ the infectiousness of phthisis. Ingestion. — A sheep fed upon potatoes which have been the medium for the cultivation of the anthrax bacillus dies in a few days. Rabbits fed on cabbage sprinkled with a culture of the bacillus of fowl cholera, rapidly succumb to the disease. Animals fed upon the nodules of bovine tuberculosis or upon tubercular flesh and milk will be readily infected. Milk, or bread soaked in milk, is a very convenient medium, and from a public health point of view, a most instructive way of administering and testing the effect of pathogenic bacteria. Vaccination and Subcutaneous Inoculation. — Vaccination may be performed by making a few superficial scratches and inoculating the wound with a sterilised platinum needle charged with the micro- organisms. Another simple method is to take a sterilised scalpel, infect the point with the material to be inoculated, and then make a minute puncture or incision. In either case a situation should be selected, such as the root of the ear, which cannot be licked by the animal after the operation. 134 EXPERIMENTS UPON THE LIVING ANIMAL. 135 Subcutaneous inoculation is very simple and effectual, and con- sequently the method most frequently employed. The animal selected — for example, a guinea-pig — is held by an assistant, who covers it with a towel, leaving only the hind extremities exposed. By so doing, and gently laying it upon its back, with its head low, a guinea-pig passes apparently into a state of hypnotism, and the FIG. 69.— KOCH'S SYRINGE. trivial operation can be performed with little or no movement on the part of the animal. From a spot on the inner side of the thigh the hair is cut close with a small pair of scissors curved on the flat, and the skin must be thoroughly purified with 1 in 20 carbolic acid. A small fold of skin is then pinched up with a pair of sterilised forceps, and with a pair of sharp sterilised scissors, or with a tenotomy knife, a minute incision is made. A sterilised platinum loop is charged with the material to be inoculated, and the loop is gently inserted under the skin, forming a small pocket in the subcutaneous tissue. The needle is then withdrawn, and the sides of the wound gently pressed into apposition and painted over with collodion. FIG. 70.— SYRINGE WITH ASBESTOS PLUG. In inoculating a mouse the same process is adopted, with the exception that the root of the tail is the usual site of the operation. In some cases it may be necessary to inoculate cultures diffused in sterilised salt solution, or blood or lymph containing bacteria, or a culture in broth, or a filtrate containing the toxic products, 136 BACTERIOLOGY. and then a hypodermic syringe may be required. One of the ordinary pattern may be used, but it is very much better to employ a syringe which has been especially constructed to admit of thorough dis- infection. Koch's syringe is a convenient form, the liquid being expressed by pressure on a rubber ball. The author has generally preferred to improvise a substitute for the hypodermic syringe which can be quickly made, and is destroyed after use, so that there can be no possible risk of accidentally infecting other animals. A short length of ordinary glass-tubing is sterilised, and plugged at one end with sterilised cotton- wool ; about three inches from the plug a bulb is blown about the size of a marble, and two inches below this the glass is drawn out into a long capillary tube. A sufficient quantity of the liquid to be injected rises up into the tube by capillary attraction, or can be drawn up by means of an india-rubber ball, until the bulb is full. The point of the capillary tube is inserted through the opening in the skin, and gently pushed into the subcutaneous tissue, and then withdrawn for a short distance. By pressure on the bulb the contents of the tube are injected. In dealing with chemical products there is no risk in applying the lips and blowing out the contents of the tube, or indeed of filling it by suction, for if too much force were applied the liquid which might enter the mouth would be stopped by the cotton-wool plug. A number of these capillary tubes can be placed in a small case, and when it is necessary to go to a distance to investigate an outbreak, they will be found most convenient to bring back lymph or blood to the laboratory for further study. Sternberg takes a piece of glass-tubing, blows a bulb at one endr and draws out the other end into a thin tube. By heating the bulb and then dipping the tube into the liquid to be inoculated the latter rises in the tube as the bulb cools. After inserting the point of the tube subcutaneously the bulb is again heated, and the liquid is forced out into the tissues. Intravenous Injection. — A cultivation of micro-organisms may be mixed with sterilised water, and then injected with a syringe directly into the circulation. This may be performed without much difficulty by injecting, with a hypodermic syringe, the large vein at the base of the ear in rabbits, or the jugular vein in large animals. Special Operations. — In many cases it is absolutely necessary to perform an operation of greater severity. After the administration of an anaesthetic, infective material may be inserted, or injected, into the peritoneal cavity, or injected into the duodenum in the manner EXPERIMENTS UPON THE LIVING ANIMAL. 137 employed in the case of Koch's comma bacilli by Nicati and Bietsch. In such cases antiseptic precautions must be rigidly followed, and use made of iodoform and other antiseptic dressings. The disinfec- tion of the skin of the animal, of the instruments employed, and of the hands of the operator, are details essential to secure success. To inoculate tubercular matter, sputum may be rubbed up with distilled water, and some of the mixture injected into a tracheal fistula ; or the first steps of the operation of iridectomy may be performed and tubercular material inserted into the anterior chamber of the eye, but this method is only justifiable when it is absolutely necessary for the results and changes to be observed from day to day. To inoculate rabbits or other animals with the virus of rabies, the skull is trephined, and an emulsion prepared from the spinal cord of a rabid animal is injected beneath the dura mater. Before every inoculation the instruments must be sterilised in a hot-air steriliser or by immersion in boiling water in a flat dish or enamel tray heated by a spirit-lamp, and after each operation all instruments should be placed in carbolic acid (1 in 20) or in boiling water, wiped dry, and again sterilised in the hot-air steriliser, before they are put away. If these precautions are not observed, instances of accidental infection are sure to occur. After the inoculation is completed a careful record must be made of the date and details of the experiment. The form in which the virus was used, the quantity employed, and the seat of inoculation, must be taken into account. The animals must be kept under close observation, the temperature taken, and any signs of illness, such as ceasing to feed, difficulty in breathing, staring coat, and any local signs, such as the development of a tumour or an enlargement of the lymphatic glands, must be carefully noted. It is perhaps hardly necessary to add that in this country no experiments of any kind may be performed on living animals without a license. METHOD OF DISSECTION AND EXAMINATION. All animals that die after an experimental inoculation should be examined immediately after death. Every precaution must be taken in conducting the dissection, to exclude extraneous micro- organisms, and all instruments employed must have been sterilised in the hot-air steriliser, or by immersion in boiling water. If a mouse, for example, has died after inoculation with anthrax, it should be at 138 BACTERIOLOGY. once pinned out by its feet on a slab of wood or in a gutta-percha tray, and bathed with 1 in 40 carbolic. In the same way, before examining a dead rabbit, a stream of carbolic should be directed, over it to lay the fur, which otherwise interferes with the dissection. The hair should be cat away with sterilised scissors from the seat of inoculation, which is the first part to be examined, and any suppuration, haemorrhage, redema, or other pathological change should be carefully noted. From any pus or exudation that may be present, material for inoculations should at once be taken, and cover-glass-preparations made for microscopical examination. To examine the internal organs and to make inoculations from the blood of the heart or spleen, the skin is cut through from below upwards in the median line of the abdominal and thoracic regions. The abdominal cavity is then opened, and the walls pinned back on either side of the animal. Any abnormal appearances in the peritoneum should be noted, and the state of the spleen should be carefully examined by turning the intestines aside. After noting its appearances, it should be removed with sterilised forceps and scissors, and deposited upon a sterilised glass slide, and incised with sterilised scissors. The cut surface is then touched with the point of a sterilised inoculating needle, and cultures are made in test-tubes of nutrient gelatine and agar-agar, and also on potato, and in broth in the form of drop- cultivations. Precisely the same care must be taken in examining lymphatic glands, tubercles, or pathological nodules ; any chance putrefactive micro-organisms on the surface should be destroyed by carbolic acid or the actual cautery; an incision is then made, and a minute fragment snipped out of the centre of the nodule, which can be inoculated in the living animal or transferred to a cultivating medium. The examination of the thorax is made by cutting through the ribs on either side of the sternum with sterilised scissors, and turning the sternum up where it will be out of the way. The pericardium is then opened, and the right auricle or ventricle pierced with the point of a sterilised scalpel, and inoculations and cover- glass-preparations are made from the blood which escapes. The lungs also require to be especially studied. They should be incised with a sterilised scalpel, and inoculations and cover-glass- preparations made from the cut surface. It may be necessary to embed a piece of lung or fragment of spleen, so that it shall be free from air. This may be done by isolating a fragment with the precautions just described, and depositing it upon the surface of a test-tube of nutrient agar-agar. The contents of another tube, EXPERIMENTS UPON THE LIVING ANIMAL. 139 which have been liquefied, and allowed to cool almost to the point of gelatinisation, must then be poured over it. From a potato a little cube must be cut, the tissue deposited in the trough thus formed, and the cube replaced, or cultures may be prepared by any of the methods which have been described for dealing with anaerobic bacteria. Blood may also be taken directly from a vein by laying it bare by dissection, making a small opening with sterilised scissors, and inserting a looped platinum needle, the needle of a hypodermic syringe, a capillary tube, or the extremity of the capillary neck of a Sternberg's bulb. If the cultivation, in spite of these precautions, is contaminated, or if there was more than one organism present in the blood or tissues under examination, it will be necessary to separate the different kinds by plate -cultivation. Having completed the dissection, the organs of such a small animal as a mouse may be removed en masse, and transferred to absolute alcohol for subsequent examination. In other cases it may be only necessary to reserve portions of each organ. In experimenting with a virulent micro-organism like anthrax, any remaining part of the animal should be cremated, and the hands and all instruments should be thoroughly disinfected. Isolation of Micro-organisms during Life. — Micro-organisms in the living subject may be isolated from the pus of abscesses, or other discharges, and from the blood and tissues. Abscesses should be opened, and other operations performed, when practicable, with Listerian precautions, and a drop of the discharge taken up with Ji looped needle or capillary pipette, as already explained. To make a cultivation from the blood of a living person, the tip of a finger must be well washed with soap and water and sponged with 1 in 20 carbolic. Venous congestion is produced by applying an elastic band or ligature to the finger, which is pricked with a sterilised sewing needle. From the drop of blood which exudes the necessary inoculations and examinations can be made. Another way of extracting blood from the living patient is to apply a leech. This method has been found of considerable value in experimenting upon the blood of patients suffering from malaria, and may be useful in other diseases, if the blood is required for further examination, or in quantity. CHAPTER XI. EXAMINATION OF AIR, SOIL, AND WATER. AlR. THE air, as is well known, contains in suspension, mineral, animaly and vegetable substances. The mineral world is represented by such substances as silica, silicate of aluminium, carbonate and phospate of calcium, which may be raised from the soil by the wind, and particles of carbon, etc., which gain access from acci- dental sources. Belonging to the animal kingdom we find the debris of perished creatures, as well as, sometimes, living animals. The vegetable world supplies micrococci, bacilli, and other forms of the great family of bacteria, spores of other fungi, pollen seeds, parts of flowers, and so forth. The air of hospitals and sick rooms has been found to be especially rich in vegetable forms; fungi and spores have been stated to be present in particularly large numbers in cholera wards; spores of tricophyton have been dis- covered in the air of hospitals for diseases of the skin, and of achorioii in wards with cases of favus. The tubercle bacillus is said to have been detected in the breath of patients suffering from phthisis. These points indicate that, in addition to the interest for the micro- biologist, considerable importance, from a hygienic point of view, must be attached to the systematic examination of the air. A knowledge of the microbes which are found jn the air of marshy and other unhealthy districts, and in the air of towns, dwellings, hospitals, workshops, factories, and mines, will be of practical value. Miquel, who has particularly studied the bacteria in the air, has found that their number varies considerably. The average number per cubic metre of air for the autumn quarter at Mont- souris is given as 142, winter quarter 49, spring quarter 85, and summer quarter 105. In air collected 2,000 to 4,000 metres above the sea-level, not a single bacterium or fungus spore was found, while in 10 cubic metres of air from the Rue de Rivoli (Paris) the number was computed at 55,000. 140 EXAMINATION OF AIR, SOIL, AND WATER. 141 The simplest method for examining the organisms in air consists in exposing plates of glass or microscopic slides coated with glycerine, or with a mixture of glycerine and grape sugar, which is stable, colourless, and transparent. Nutrient gelatine spread out on glass plates may be exposed to the air for a certain time, and then put aside in damp chambers for the colonies to develop. Sterilised potatoes, prepared in the usual way, may be similarly exposed. In both the last- mentioned methods separate colonies develop, which may be isolated, and pure cultivations carried on in various other nutrient media. Nutrient gelatine has also been employed in the special methods of Koch and Hesse. Koch's Apparatus. — This consists of a glass jar, about six inches high, the neck of which is plugged with cotton-wool. In the interior is a shallow glass capsule, which can be removed by means of a brass lifter. The whole is sterilised by exposure to 150° C. for an hour in the hot-air steriliser. The nutrient gelatine in a stock-tube is liquefied, and the contents emptied into the glass •capsule. The jar is exposed to the air to be examined for a definite time, the cotton -wool plug replaced, and the apparatus set aside for the colonies to develop. Hesse's Apparatus. — The advantage of this apparatus is that it enables the experimenter to examine a known volume of air. A glass cylinder, 70 cm. long and 3*5 cm. in diameter, is closed at one end by an india-rubber cap, perforated in the centre. Over this fits another cap, which is not perforated. The opposite end of the cylinder is closed with a caoutchouc stopper, perforated to admit a glass tube plugged with cotton-wool. The tube can be connected by means of india-rubber tubing with an aspirating apparatus, which consists of a couple of litre-flasks, suspended by hooks from the tripod-stand which supports the whole apparatus. The cylinder, caps, and plug are washed with solution of carbolic acid, and then with alcohol. After being thus cleansed, 50 cc. of nutrient gelatine are introduced, and the whole sterilised by steaming for half an hour for three successive days. After the final sterilisation, the cylinder is rotated on its long axis, so that the nutrient medium solidifies in the form of a coating over the whole of the interior. When required for use, the cotton-wool plug is removed from the small glass tube, and the latter connected with the upper flask by means of the india-rubber tubing. The apparatus is placed in the air which is to be examined, the outer india-rubber cap removed from the glass cylinder, and the upper flask tilted until the water begins to flow into the lower one. 142 BACTERIOLOGY. The emptying continues by siphon action, and air is drawn in along the cylinder to replace the water. When the upper flask is empty, the position of the two is reversed, and the flow again started. When a sufficient volume has been drawn through the cylinder, the outer cap and the cotton-wool plug are replaced, and it is set aside for the colonies to develop. As an example, twenty-five litres of air from an open square in Berlin gave rise to three colonies of bacteria and sixteen moulds; on the other hand, two litres from a school - FIG. 71.— HESSE'S APPARATUS. • room just vacated by the scholars gave thirty-seven colonies of bacteria and thirty-three moulds. Porous substances, such as sand, powdered glass, or sugar, may be used for the filtration of samples of air ; and an apparatus is employed in a convenient form to be conveyed to the laboratory for the subsequent examination. Petri's Apparatus consists of a glass-tube 9 cm. long, containing two sand-filters separated from each other. A known volume of air is aspirated through the tube. The bacteria are arrested and can EXAMINATION OF AIR, SOIL, AND WATER. 143 be examined by spreading the sand out in a dish and covering it with nutrient gelatine ; or it may be shaken up with sterilised water and pi ate -cultivations prepared. The sand-filter nearest to the aspirator should remain free from bacteria. Sedywick and Tucker employ a glass cylinder which is drawn out at one end into a narrow tube to contain sterilised powdered cane sugar. Both ends of the apparatus are plugged with sterilised cotton-wool. By means of an exhausting apparatus a known volume of air is drawn through the tube. The cotton-wool plug is re- FIG. 72. — SEDGWICK AND TUCKER'S TUBE. moved, and liquid gelatine is introduced into the cylinder, the plug is replaced, and the sugar is shaken into and quickly dissolves in the jelly. The cylinder is then treated in the same way as a roll-culture, and set aside for the colonies to develop (Fig. 72). Various forms of " aeroscopes " and " aeroniscopes " have from FIG. 73.— POUCHET'S AEROSCOPE. time to time been employed. Pouchet's aeroscope consists of a small funnel, drawn out to a point below which is a glass slip coated with 144 BACTERIOLOGY. glycerine. The end of the funnel and the glass slip are enclosed in an air-tight chamber, from which a small glass tube passes out and is connected by india-rubber tubing with an aspirator (Fig. 73). The air passing down the funnel strikes upon the glycerine, which arrests any solid particles. For a full description of the apparatus employed by Maddox, Cunningham, and Miquel, reference should be made to the writings of these authors, and particularly to the treatise published by the last-named. SOIL. Surface soil is exceedingly rich in bacteria. Miquel has com- puted that there exists in a gramme of soil an average of 750,000 germs at Montsouris, 1,300,000 in the Rue de Rennes, and 2,100,000 in the Rue de Monge. As agents in putrefaction and fermenta- tion they play a very important rdle in the economy of nature ; but there exist in addition, bacteria in the soil which are patho- genic in character. Pasteur has succeeded in isolating the bacillus of anthrax from the earth. Sheep, sojourning upon a plot of ground where animals with anthrax have been buried, may succumb to the disease. Pasteur considered that the spores were conveyed by worms from buried carcasses to the surface soil. The bacilli of malignant redema and tetanus are also present in soil. Nicolaier produced tetanus in mice and rabbits by inoculating a little garden earth under the skin. To obtain a cultivation of the microbes in soil a sample of the latter must be first dried and then triturated. It may then be shaken up with distilled water, and from this a drop transferred to sterilised broth. The employment of solid media is, however, much more satisfactory : a sample of earth is collected, dried, and triturated, and a small quantity sprinkled over the surface of nutrient gelatine prepared for a plate-cultivation. In another method the gelatine is liquefied in a test-tube, the powder added, and distributed, in the usual way, throughout the medium, which is then poured out upon a glass plate or made into a roll-culture. In the same way the dust which settles from the air in houses and hospitals, or food substances in powder, may be distributed in nutrient gelatine, and examined both for aerobic and anaerobic bacteria. The different kinds which develop, must be thoroughly investigated as regards their morphological and biological charac- ters, and pathogenic properties. EXAMINATION OF AIR, SOIL, AND WATER. 145 WATER. In the case of water, as in that of air, a knowledge of the micro-organisms which may be present is not only of interest to the bacteriologist, but of the greatest importance in practical hygiene. Common putrefactive bacteria and vibrios may not be hurtful in themselves, but they indicate the probability of the presence of organic matter in which there may be danger. The detection of Bacillus coli communis may be taken to indicate a probable contamination with human excreta. The Microzyme Test which was introduced for the detection of putrefactive bacteria, consisted in adding three or four drops of the sample of water to 1 or 2 cc. of Pasteur's fluid, the nourishing fluid having been previously boiled in a sterilised test-tube. If the microzymes or their germs existed in the water, the liquid in a few days became turbid from the presence of countless bacteria. This test is of 110 real value, for it does little more than indicate that bacteria are present, which we know to be the case in all ordinary water, and even in ice. On the other hand, the bacteriological test of Koch is a most valuable addition to the usual methods of water analysis. It enables us not only to detect the presence of bacteria, but to ascertain approximately their number, and to study very minutely their morphological and biological characteristics. The importance of a thorough acquaintance with the life- history of the individual micro-organisms cannot be too strongly insisted upon. For example, by such means the spirillum of Asiatic cholera can be distinguished from most other comma-shaped organisms, and inasmuch as its presence may be an indication of contamination with choleraic discharges, such water should be condemned for drinking purposes, even though we are not yet in a position to affirm that the microbe is the cause of the disease. The detection of the bacillus of typhoid fever or of the Bacillus coli communis in suspected water or milk would be evidence of considerable importance. Koch's test, in short, consists in making plate-cultivations of a known volume of water, counting the colonies which develop f isolating the micro-organisms, and studying the characters of each individual form. Collection and Transport of Water Samples. — Sternberg's bulbs, or Erlenmeyer's conical flasks of about 100 cc. capacity, may be employed with advantage for collecting the samples of water. The latter are cleansed, plugged, and sterilised in the hot-air steriliser. 10 146 BACTERIOLOG Y . When required for use, the plug is removed and held between the fingers, which must not touch the part which enters the neck of the flask. About 30 cc. of the water to be examined are intro- duced into the flask, and the plug must be quickly replaced and covered with a caoutchouc cap. If collected from a tap, the water should first be allowed to run for a few minutes, and the sample should, be received into the flask without the neck coming into contact with the tap. From a reservoir or stream, the flasks may be filled by employing a sterilised pipette. During transport contact between the water and cotton-wool plug must be avoided, and if likely to occur the sample must be collected and forwarded in a Sternberg's bulb. Examination ~by Plate-cultivation. — The apparatus for plate- cultivation should be arranged as already described. Crushed ice FIG. 74. — APPARATUS FOR ESTIMATING THE NUMBER OF COLONIES IN A PLATE-CULTIVATION. may be added to the water in the glass dish to expedite the setting of the gelatine, so that the plate may be transferred as quickly as possible to the damp chamber. The caoutchouc cap is removed from the flask, and the cotton- wool plug singed in the flame to prevent contamination from adventitious germs on the outside of the plug. The flask is then held slantingly in .the hand, and the plug twisted out and retained between the fingers. With a graduated pipette a measured quantity (y1^ or -^ cc.) of the sample is transferred to a tube of liquefied nutrient gelatine, and the plugs of the flask and of the tube quickly replaced. If the water is very impure, it may be necessary to first dilute the sample with sterilised water. The inoculated tube must be gently inclined backwards and forwards, and rolled as already explained, to distribute the germs throughout the gelatine, and the gelatine finally poured on a plate. When the gelatine has set, the plate is transferred to a damp chamber, which should be carefully labelled and set aside in a place EXAMINATION OF AIR, SOIL, AND WATER. 147 of moderate temperature. In about two or three days the cultivation may be examined. In some cases the colonies may be counted at once ; more frequently they are so numerous that the plate must be placed on a dark background, and a special process resorted to. A glass plate, ruled by horizontal and vertical lines into centimetre .squares, some of which are again subdivided into ninths, is so arranged on a wooden frame that it can cover the nutrient -gelatine plate without touching it (Fig. 74). A lens is used to assist in dis- covering minute colonies. If then the colonies are very numerous, the number in some small division is counted, if less in some large one, and an average is obtained from which the number of colonies on the entire surface is calculated. A separate calculation of the liquefied colonies should be also made, and their number, as well as the total number of colonies present in 1 cc. of the sample, recorded. Any peculiar macroscopical appearances, colours, etc., should be noted, and then the microscopical appearances of the colonies studied. Lastly, examination of the individual organisms should be made by cover-glass preparations, and by inoculation of nutrient gelatine, potatoes, and other media. Instead of plates, Petri's dishes may be used both for gelatine .and agar-agar cultivations. FIG. 75. — ESM ARCH'S ROLL-CULTURE. .«, India-rubber caps; b b b, longitudinal line drawn on the tube; c,c,c, transverse lines for counting colonies (FRANKLAND). Another plan is to take a measured quantity of the sample of water and prepare a roll-culture, using a large-sized test-tube (Fig. 75). The colonies can be counted with the aid of a lens (Fig. 76). Microscopical preparations and sub-cultures can be made from the colonies, and the anaerobic bacteria can be examined by Frankel's modification of this method (p. 131). A drop of the sample of water may also be added to liquefied nutrient gelatine in a test-tube, the organisms distributed, and the gelatine allowed to solidify in the tube. A rough comparison of water samples may be made in this way. Microscopic Examination. — A drop of the water may be mounted and examined without staining ; or allowed to evaporate on a cover- 148 BACTERIOLOGY. glass, which is then passed through the flame, and stained in the- usual manner. Parietti's Method. — As typhoid fever bacilli are apt to be crowded out by more rapidly growing micro-organisms, some method had to be devised for restraining the growth of the latter, and Chantemesse and Widal suggested the use of carbolic acid. Parietti put this into practice by the method he introduced. This consists in adding to tubes of broth about five drops of a mixture composed of sterilised water (100 parts), hydrochloric acid (4 parts), and carbolic acid (5 parts). The tubes are first tested by incubation, and are then ready for use. A few drops of the suspected water are added FIG. 76.— APPARATUS FOR COUNTING COLONIES IN A ROLL CULTURE. to the broth, and if it becomes turbid in a day or two the typhoid1 fever bacillus is present in the form of a pure- cult tire. An excess of bacteria in a fresh sample indicates an excess of organic matter, and points to possible contamination with sewage. Where there is such contamination we are very likely to find pathogenic bacteria ; and moreover impure water is a constant source of danger, for if the contagia of infectious diseases ar& introduced they will retain their vitality in such water for a long period, and will in some cases even multiply, whereas the same organisms introduced into pure water would in a short time perish. The actual number of bacteria in water is not of very great importance, and it must be remembered that if a sample is set aside for a few days there will be an enormous increase in the number of bacteria present ; but in dealing with perfectly fresh samples it EXAMINATION OF AIR, SOIL, AND WATER. 149 may be said that water containing less than 100 bacteria to the cubic centimetre is very pure water. Water containing 1,000 or more should be filtered. Water containing 100,000 to 1,000,000 is contaminated with surface water or sewage. It is necessary to bear in mind that in typhoid fever and Asiatic cholera the excreta contain the bacteria in great numbers, and wells and streams receiving surface water may be contaminated in various ways. The cholera bacillus dies as a rule quickly in distilled water, while it preserves its vitality for a long time in water of a bad quality. It is necessary to lay stress upon the fact that a bacterio- logical analysis may show the presence of pathogenic bacteria when their detection is not possible by any other means. They may be present in water in such small numbers that no chemical analysis would detect any contamination, but as they are living organisms capable of increasing in a suitable environment, they can readily be discovered by bacteriological methods. The examination of rain water, drinking water, tap water, sea water, various liquids and infusions, by these methods, opens up a wide field for research. Pettenkofer has shown that impregna- tion of water containing many bacteria with carbonic acid diminishes the number of the latter. The examination of waters before and after filtration, or after addition of chemical substances, are matters which require further investigation, though a great deal of work has already been accomplished. The reader will find in Micro-organisms in Water by P. and G. Frankland, a very complete account of this subject with valuable analytical tables. CHAPTER XII. PHOTOGRAPHY OF BACTERIA. THE production of pictures of microscopic objects by photographic means was attempted at an early date. Some authorities regard the very earliest recorded experiments as being the first experi- ments alike in photography and micro-photography. The experi- ments of Wedgwood and Sir Humphry Davy were embodied in a paper read before the Royal Institution in 1802. They obtained with the solar microscope impressions upon paper, and with greater success upon white leather, though the results were transitory when exposed to daylight. In 1816 Mcephore Niepce described his experiments in con- nection with fixing the image obtained by the camera. He was at first only able to obtain negatives, and these were transitory. But, after joining with Daguerre, who had been experimenting in the same direction, a process was invented which was published in 1839 under the name of daguerreotype. This invention, and the rapid improvements which followed, were taken advantage of by Reade, Donne, Hodgson, Kingsley, and Talbot, who were early workers in the field of micro-photography. So early as 1845 it is stated that Donne produced a work illustrated with engravings copied from daguerreotypes. Subsequently this interesting branch of photography was taken up by many in France and Germany, in America, and in England. Of those to whom we are indebted for the literature of the subject, and for many improvements, the names of Wenham, Dancer, Draper, Maddox, Shadbolt, Redmayne, Woodward, Highley, Deecke, Moitessier, Gerlach, Koch, Sternberg, Frankel, Pfeiffer, and Pringle may especially be mentioned. Of these workers the name of Woodward stands pre-eminently foremost. His skill in microscopical manipulations, combined with access to- the very best apparatus and objectives, placed at 150 PHOTOGRAPHY OF BACTERIA. 151 his disposal in the Museum at Washington, enabled him to obtain photographs of diatoms which probably have never been surpassed. To Koch belongs the credit of being the first to extend the application of micro-photography to the delineation of bacteria. A series of instructive photographs was first published by him in 1877. These were photographs of cover-glass preparations, and all admirably illustrated the subjects from which they were taken ; while two, showing the flagella of bacilli and spirilla, were triumphs in this new departure. Lewis, in India, was one of the first to illustrate his writings on the subject of micro-organisms by means of photographs. About the same time Sternberg, in America, took some excellent photographs of bacteria. Heliotypa reproductions of these were published in 1884. Hauser and Van Ermengem and many other bacteriologists successfully resorted to photography for illustrating their researches, and Frankel and Pfeiffer's, and Itzerott and Niemann's atlases of photographs of bacteria, in microscopical specimens and cultivations, are especially worthy of mention. Opinions have differed widely as to the merits of photographic illustrations. Many, taking the standpoint solely of a comparison with drawings, have decried their use. By judging from such a comparison alone the real value of photographs may be lost sight of. On the other hand, many who have looked at the question from all sides, have been led to value even a defective photograph more than an ordinary drawing. In his first publication on this subject, Koch strongly advocated photography on the ground that illustrations would then be as true to nature as possible. The photographs which accompanied his paper were all taken from preparations of bacteria which had been made from blood, cultivations, or infusions, by drying a thin layer on a cover-glass and staining, or from specimens prepared in the same way but left unstained. But when, having committed himself to this opinion, Koch attempted, later, to photograph the bacteria in animal tissues, he was led to modify his previous conclusion. For though no trouble was spared, yet disappointing results were met with. This was owing, he explains, to the fact that the smallest and most interesting bacteria can only be made visible in animal tissues by staining them, and thus obtaining the advantage of colour. This introduced the same difficulties which are met with in photographing coloured objects, such as tapestry and oil paintings. 1 52 BACTERIOLOGY. As these difficulties had been to a certain extent obviated by the use of eosin-collodion, Koch adopted the same method for photo- graphing stained bacteria. By the use of eosin-collodion, and by shutting off portions of the spectrum by coloured glasses, he succeeded in obtaining photographs of bacteria which had been stained with blue and red aniline dyes. But, owing to the long exposure which was necessary, and the unavoidable vibrations of the apparatus, the results were so wanting in definition that they not only proved unsatisfactory as substitutes for drawings, but did not in some cases give any evidence of what was to be seen in the preparations. Koch, in consequence, stated that he would abstain from publishing photographic illustrations until he had the advantage of improved methods. We find, however, in spite of this, that in 1881 Koch published a series of reproductions from his negatives in illustration of what could be accomplished by photography. Here again we find that many of the photographs of cover- glass preparations were admirable, but those of tissue-sections gave evidence of the difficulties Koch encountered, and were undoubtedly unsatisfactory from the want of flatness of field, some of the illustrations recalling rather a map of a mountainous country than a microscopical preparation. In consequence of the difficulties met with in attempting to photograph bacteria stained with the aniline dyes most commonly used, Koch recommended that the preparations should be stained brown, pointing out as his reason that, though the bright and concentrated colour of the red and blue aniline dyes catches the eye far more readily than the somewhat sombre brown colours, yet no one up to the time of his publication had succeeded in obtaining good photographs of bacteria which had been stained either blue or red, and mounted in Canada balsam, while there was no difficulty in obtaining photographic representations of prepara- tions stained yellow or brown. Though this stain could be easily employed in most cover-glass preparations, it was by no means easy to obtain a good differential stain of bacteria in the tissues by employing Bismarck brown. An attempt was, therefore, made to photograph preparations stained blue and red by the aid of the dry-plate process, and by interposing glasses of suitable tints. After many fruitless experi- ments this method had to be abandoned, arid the method of staining the object brown was adopted. In many cases this gave excellent PHOTOGRAPHY OF BACTERIA. 153 results ; in others again, compared with the results of staining with blue or red stains, there was much to be desired, and further improvement was needed. That a stain, such as yellow or brown, must be employed which absorbs the blue rays, and acts on the sensitive plate like black, which absorbs all the light, constituted the first condition laid clown by Koch as an essential for success. It was further pointed out that the suitability of the stain could be ascertained by first passing the light, to illuminate the preparation, through a solution of ammonio- sulphate of copper, under which condition the bacteria would appear black on a blue ground. The second condition was, that sunlight must be employed, but that direct projection upon the object was disadvantageous, and it must, therefore, be diffused by the interposition of one or more plates of ground glass. Lastly, an illuminating condenser was recommended, of such construction that the diffused sunlight brightly illuminates the object from all sides. Sternberg encountered the same difficulty in photographing red, blue or violet preparations, while he produced excellent pictures of preparations stained with aniline brown, or a weak solution of iodine (iodine grs. iij, potassic iodide grs. v, distilled water grs. 200). Thus the results of a large number of attempts to photograph the tubercle bacillus in sputum, only ended in producing such extremely faint impressions, that any one unacquainted with the object as seen under the microscope could form scarcely any idea of its form or minute structure with even an accompanying explanation and the closest inspection of the photograph. Dufrenne, in attempting to photograph the same object by the ordinary method, found the plates were uniformly acted on, or the image was so faint, or so lacking in contrast, that they were useless for obtaining proofs on paper or glass. By interposing green glass between the objective and the sensitive plate, so that the red rays were absorbed, while the green rays passed through and acted on the plate, he states that better results were obtained. The work of Hauser illustrated the great value of photography in the production of pictures of impression-preparations and colonies in nutrient gelatine. To give the general effect, as well as faithfully reproduce the minute details in these difficult subjects would in most cases create insurmountable difficulties, except to the most accom- plished draughtsman. Hauser employed Gerlach's apparatus and Schleussner's dry 154 BACTERIOLOGY. plates, and obtained the illumination by means of a small incan- descent lamp, which gave a strong, white light, with three or four Bunsen elements. In another respect Hauser's results were of practical value. The preparations to be photographed were stained brown as recommended by Koch, but they were mounted in the ordinary way in Canada balsam. The objection to the mounting medium most commonly employed was thus set aside. The prevalent idea, however, that the preparations must be stained brown was still a formidable obstacle, and the way out of this difficulty was clearly- shown by Yan Ermengem's photographs. These were pictures of comma-bacilli which had been stained with fuchsine and methyl violet. These photographs afforded the first practical illustration of the value of isochromatic plates in micro-photography which had been previously noted by Yan Ermengem in 1884, and their intro- duction marks a distinct era in the progress of micro-photography. A short explanation may be given of what is meant by isochro- matic, or what have been more properly termed orthochromatic dry plates. The difficulties encountered in photographing certain stained preparations have been mentioned.1 It is a familiar fact that in portraits, blue or violet comes out almost or quite white, while other colours, such as yellow, are represented by a sombre shade or perhaps black. This failure in correctly translating colours is explained by the want of equality between the strength of the chemical and luminous rays. If the rays of the spectrum are pro- jected upon a photographically sensitive surface, the greatest effect is found to take place at the violet end. In other words, the violet and blue rays are more actinic or chemically powerful, while the yellow and orange have scarcely any effect. The dyes employed in staining give corresponding results : blue and violet give but a faint impression, yellow and orange a black picture. These results are most clearly demonstrated in a photograph of an oil painting taken in the ordinary way ; and they led to experiments being made which have resulted in orthochroinatic photography. The effect of interposing coloured glasses has already been referred to. It was found later that, if plates were coloured yellow, e.g., with turmeric, the blue and violet rays were intercepted, and their actinism reduced. In 1881, Tailfer and Clayton produced the so-called isochromatic plates. The emulsion of bromide of silver and gelatine was stained with eosin, and it was claimed that colours would be represented with their true relative intensity. Chlorophyll and other stains have been tried, and by such methods the ordinary gelatine dry plates can be so treated that they will reproduce PHOTOGRAPHY OF BACTERIA. 155 various colours, according to their relative light intensity, and thus be rendered iso- or, what is now more commonly known as, ortho- chromatic. APPARATUS AND MATERIAL. Micro-photographic Apparatus. — As is well known, various forms of apparatus have from time to time been recommended and em- ployed by different workers. Many use the microscope in a vertical position, with the camera superposed or fitted to the eye -piece end of the microscope tube ; or the microscope tube may be screwed off from the body of the microscope, and a pyramidal camera adjusted in its place, the base of the pyramid being represented by the ground glass screen. Others again prefer that the microscope and camera should be arranged horizontally. In another form the ordinary microscope is dispensed with, the objective, stage, and mirror are adapted to the front of the camera, and provided with suitable arrangements for holding the object, supporting the mirror, and adjusting the different parts. Lastly, the camera may be dispensed with, the operating-room, which must be rendered impervious to light, taking its place, while the image is projected and focussed upon a ground glass screen, which has a separate support.* The horizontal position affords greater stability than the vertical, so that the former is to be preferred. The vertical model with the camera fixed to the microscope is particularly to be avoided, as the weight of the camera bears directly upon the microscope, and must affect the fine adjustment, and any vibration in one part of the apparatus is communicated throughout. The simplest apparatus consists of a camera fixed upon a base- board four or five feet in length, upon which the microscope can be clamped, and which carries also a lamp and a bull's-eye condenser (Fig. 77). Simplicity and economy must always be borne in mind in recommending any apparatus of this kind, for to insist upon the necessity of a very elaborate apparatus, or a specially fitted- up room, or that a special room should be built with windows facing in a definite direction, will in most cases at once place photography beyond the reach of those who might otherwise employ it. Yet to fulfil * For an excellent account of the forms of apparatus which have been employed by different workers the reader is referred to the section on Micro- photography in Beale's How to work with the Microscope. 156 BACTERIOLOGY. all the purposes for which the apparatus may be required, including the employment of the highest powers, and also that one may be enabled to work for long intervals of time with due comfort, an accurate and complete apparatus will be found to be most desirable. Though most preparations will admit of being photographed when the stage of the microscope is vertical, yet if we require to photograph micro-organisms in liquids, or colonies upon partially liquefied gelatine, the apparatus must admit of being placed so that the stage of the microscope becomes horizontal. In addition, the apparatus is rendered somewhat complex if we employ powerful FIG. 77. — HORIZONTAL MICRO-PHOTOGRAPHIC APPARATUS. artificial light. Sunlight, no doubt, is the best and cheapest, but it is not always available, especially in a city like London; and, moreover, evenings and dull days will probably be the very time which can be best spared for this work. We must, therefore, fall back upon the paraffine lamp, or the magnesium, oxyhydrogen, or electric light. To fulfil all these conditions Swift has constructed an apparatus under the author's directions (Fig. 78). It is merely a modification of the ordinary horizontal model, which admits of being readily placed PHOTOGRAPHY OF BACTERIA. 157 in the vertical position, while the illumination is supplied from an oxyhydrogen lantern. To place the apparatus in the vertical position two small hinged brackets, at the end distant from the camera, are forced up with a smart blow of the hand. The corresponding ends of the stretcher bars are dislodged from their fittings, and allowed to descend ; when 158 BACTERIOLOGY. horizontal, the opposite extremities of the bars are easily released from their sockets. The leg or support at this end can then be FIG. 79. — REVERSIBLE MICRO-PHOTOGRAPHIC APPARATUS ARRANGED IN THE VERTICAL POSITION. turned up and fixed underneath the apparatus by a button, and the end of the apparatus itself gently lowered to the ground. PHOTOGRAPHY OF BACTERIA. 159 A hinged end-piece is also to be turned out to increase the base upon which the whole apparatus will stand when raised to the vertical. The two-legged support at the opposite end of the apparatus is next worked down by a quick thread screw, and on raising the apparatus to the vertical, the two-legged support drops to the ground, and assists in maintaining the stability of the whole. If it is thought necessary, a simple means can be readily devised for clamping the apparatus, in either position, to the wall of the room, so as to eliminate as much as possible all chances of vibration. A second quick thread screw moves the base-board upon which the camera and central sliding-board are mounted, so that the camera, microscope and lantern can be raised to a convenient height from the ground. The various parts of this apparatus may be described in detail. The Microscope and its Attachments. — It is most essential that the microscope should be perfectly steady. The microscope was made by Zeiss, and to ensure steadiness, the horse- shoe foot piece tits under a projecting ledge, and is then clamped by a cross-piece, so that it is firmly fixed. The microscope with the means for clamping it and the oxy- hydrogen lantern are carried upon an independent sliding-board, which admits of movement to or from the camera. The sliding- board also moves upon a centre, which enables the microscope to be turned out from the median line ; in fact, to be turned at a right angle to the position it occupies when ready for the exposure. The object of this contrivance is to enable the operator to sit down by the side of the apparatus, and with comfort to arrange the object in the field of the microscope. On turning the microscope back into the median line, it is fixed in the optical axis of the apparatus by means of a suitable stop. The sliding-board is provided with a small grooved wheel receiving an endless cord, made of silk or fishing-line, which passes round the grooved, milled head of the fine adjustment of the microscope. When the sliding-board is returned to the median line of the apparatus, the milled wheel connected with the fine adjustment impinges upon the wheel of the long focussing rod. The latter is provided with an india-rubber tire, which grips the teeth of the milled wheel, and thus the long focussing rod is placed in connection with the fine adjustment of the microscope. Illumination. — The oxy-hydrogen lamp has been more frequently employed by the author than the parafnne lamp, partly on account of the diminished time in exposure, especially when employing very ^-5w^ *? KN. OF TRR X UNIVERSITY ] °f rTT^ ui 160 BACTERIOLOGY. PHOTOGRAPHY OF BACTERIA. 161 high powers ; this is of great importance where there is likely to be vibration from passing traffic. With rapid plates and the highest powers, the exposure has only been two or three seconds, whereas with the paraffine lamp it may vary from three to ten minutes, or even longer. Walmsley gives the following table for exposures with the paraffine lamp : — 1| inch objective . . . . 3 to 45 seconds. 7 on 3 » ,, . . . . l ,, v\J ,, •=ir i . , 3 minutes. l 9 -§ » » . . . . Z i 4. 1 0 10" " " " " The illuminating apparatus represented in the accompanying engraving (Fig. 78) consists of a lantern which not only moves together with the microscope on the central sliding-board, but can be moved independently to or from the microscope, and be clamped with screws at the requisite distance for obtaining the best illumination. It is provided with two 3-inch condensing lenses of long focus, constructed of optical glass, which is much whiter than that used for ordinary lantern condensers. The lime-cylinders should be of the hardest and best quality, as they give a more actinic light than those made of soft lime. The " Excelsior " lime-cylinders are strongly recommended. They are supplied in hermetically sealed tins which can be easily opened and re-sealed, so that a cylinder can be taken out and used, and the rest preserved for a future occasion. The hydrogen can be obtained by using the coal-gas supplied to the house, and the oxygen should be supplied preferably in a compressed state in iron bottles. Not only are the bottles much less cumbrous than the bags, but a small quantity of gas can be used, and the residue left for an indefinite time ; moreover, the gas is always at hand to be turned on when required. On the other hand, the retention of unused gas in bags is liable to cause their corrosion, owing, it is believed, to impurities which are carried over in the manufacture of the oxygen. If gas is not laid on in the house, then it also must be procured in a compressed state in bottles. As the blow-over jet is recommended on account of its safety, the bottles should be supplied in this case with a supplementary valve. It is then just as easy and free from danger to employ the compressed gas as it is to make use of the house-supply. The Camera. — A long-focus, half-plate camera is mounted upon a sliding platform. This admits of the camera being pushed up to 11 162 BACTERIOLOGY. the microscope when it is in the long axis of the apparatus, so as to make a light-tight combination. The opening which is filled in an ordinary camera by the lens can be shut off by means of an internal shutter, which is opened and closed by turning a screw at the side of the camera. The dark-back is provided with plate-carriers, so that either half, quarter, or lantern-size plates can be employed. It will be found convenient to have two or more dark- backs, so that several plates may be exposed without rearranging the light for each exposure. Much more elaborate and expensive micro- photographic cameras have been constructed by Zeiss, and also by Swift. The latter has FIG. 81.— PHOTOGRAPH OF AN IMPRESSION PREPARATION. carried out a suggestion made by Pringle for a support at the ocular end (Fig. 80). The Dark-room. — In every bacteriological laboratory there should be a developing room provided with shelves, gas, water-tap, and sink, but these arrangements are not absolutely indispensable. All that is essential is a room impervious to light ; and a closet or cupboard, if it can be ventilated, will answer perfectly well, with a jug and basin instead of the tap and sink. The steam-steriliser employed in the preparation of nutrient media for cultivating bacteria, if not required at the time for such purposes, may be filled to the brim with water, and will form an excellent cistern and tap, while a pail, or small sanitary bin, may be utilised as a sink. Various kinds of lamps are made for the dark-room, burning PHOTOGRAPHY OF BACTERIA. 163 either candles, oil, or gas. In any case, the light must pass through two thicknesses of ruby glass. Dry Plates. — A small supply of any of the ordinary plates in the market may be procured for preliminary trials in acquiring a knowledge of the processes ; but to overcome the difficulties of certain stained preparations, the isochromatic or orthochromatic plates should be used. The | plate will be found to be the most suitable size. There are numerous formulae for the requisite solutions for developing and fixing the negatives, and instructions are usually enclosed in the boxes of dry plates, but it is best to abstain from trying a number of different formulae, as it leads to a great expendi- ture of time. There is a temptation to do this, it being supposed that there is probably some great advantage in one formula over another. It is much better to get accustomed to the behaviour under different exposures of one, or perhaps two methods. In France the iron developer is much in vogue, and is recom- mended by Tailfer and Clayton for use with their isochromatic plates. It has the advantage of great simplicity in the mode of employment, and, therefore, is very suitable for a beginner. In England, on the other hand, the alkaline developer is very much used, as it gives more command over the plate, enabling the photographer more fully to compensate for incorrect exposure. It is very desirable before attempting to take photographs with the microscope to learn how to take photographs with an ordinary landscape camera, and to get thoroughly accustomed to the use of some good developer, so that mistakes may be corrected and the clearest and sharpest negatives obtained. PRACTICAL MANIPULATION. Arrangement of Apparatus. — For working with the paraffiiie lamp, the mode of procedure is, as regards the illumination, briefly as follows. The sub-stage condenser is dispensed with when a low power is employed, as well as the mirror, and the lamp is so placed that the image of the flat of the flame appears accurately in the centre of the field of the microscope. A bull's-eye condenser is then interposed, so that the image of the flame disappears, and the whole field is equally illuminated. With high powers the sub-stage achromatic condenser is necessary, and a more intense illumination is obtained by using the flame edgewise. In using a low power with the oxyhydrogen light, the lantern is withdrawn some little distance from the microscope, and the top combination of the achromatic condenser removed. 164 BACTERIOLOGY. It is best to begin with the use of a low power, and a trial object, such as the blow-fly's tongue, spine of Echinus, or trachea of silkworm. In order to explain the management of the apparatus (as represented in Fig. 78) the steps in the arrangement of the apparatus and exposure of the plate will be described in detail for the employment of a high power and the oxyhydrogen light. The solutions being ready for use, it is proposed to take a photograph of tubercle bacilli in sputum, with a -f% apochromatic oil-immersion objective. The first point to claim attention is the arrangement of the light. Having lighted the gas at the hydrogen jet, the lime-cylinder should be revolved until heated equally all round. The oxygen is then carefully turned on until only a small spot of incandescence is produced. The central sliding-board is turned out, a low power screwed on to the microscope, and the image of the bright spot focussed and accurately centered. To protect the sight, an eye-pieca provided with a smoked glass shade is used. The immersion objective is then substituted for the low power, and the oxygen turned on until the right admixture of gas is obtained to produce a brilliant illumination. It is well at this stage to sit down to focus the selected object, and to spend some little time in searching for the most characteristic part of the specimen to be photographed. This being decided upon, the eye- piece is carefully withdrawn, and the central sliding-board rotated back into the median line. To make a light-tight connection between the camera and the microscope, the camera is pushed up until a velvet-lined tube, which occupies the position of the lenses of an ordinary camera, is enclosed within a short wide tube which is adapted to the eye-piec3 end of the microscope. On opening the camera-shutter the image will be projected upon the ground glass screen of the camera. It is necessary, however, to obtain the exact focus, and to effect this the ground-glass screen is turned away, and the dark-back with a piece of plain glass is substituted. Here again time may be well spent in getting the sharpest image, with the aid of a focussing glass of proper focal length. The greatest delicacy in manipulation is necessary, as in working with such high powers a turn too much of the fine adjustment will cause the image to vanish. Having determined the best visual focus, which will be found with the high-power objectives of most makers to correspond with the chemical focus, the dark-back must be cautiously removed, to prevent any vibration, and the plain PHOTOGRAPHY OF BACTERIA. 165 glass replaced by a sensitive plate. To effect this change, the operator retires to the dark-room, and opens a box of plates with as little exposure to the red light as possible. Having removed a plate, it is necessary to ascertain which is the sensitive side* This may be done by momentarily exposing it to the red light, and seeing which is the sensitive side by the dull appearance of the film. A less satisfactory way is to moisten the tip of a finger, and press it at one corner of the plate. The film side will be recognised by imparting a sticky sensation. The film must be dusted with a camel's- hair brush, as well as the dark- back, and the plate is placed film-side downwards in the dark- back, which is then securely closed. Care should be taken that the plates then remaining in the box are packed away before light is admitted to the dark-room. Exposure of the Plate. — On returning to the apparatus, the camera-shutter is closed. Then the dark-back is gently slid into its place, and its slide withdrawn. A few moments are allowed to elapse, so that tne least possible vibration, which might be caused by inserting the dark- back, has had time to cease, and all is ready for the exposure. In the case of the object we have selected, three seconds will probably be the exposure required. This is done by opening and closing the earner a -shutter with one hand, while a watch can be held in the other. The slide of the dark-back is then carefully closed, and the plate is ready to be carried off to the developing room. As the light will not be again required until the next exposure, the oxygen must be turned off, while the coal-gas may be allowed to play over the lime. Development and Fixation of the Image. — It is well to be systematic, and * therefore, before the plate is taken out of the dark-back, light is admitted to the dark-room, and everything arranged so that the position of the trays and bottles may be remembered in the dark. First, let the ruby lamp be lit, place two dishes or trays close by, and a row of four dishes within easy reach. Pour out some fixing solution in the first porcelain dish, alum in No. 2, and water in Nos. 3 and 4. Put the necessary quantity of " pyro " solution into the glass measure, and place it with the ammonia drop-bottle in front of the ruby light. Then, when all light except that from the ruby lantern has been excluded, everything is ready to commence the development of the plate. 166 BACTERIOLOGY. Opening the dark-back, the plate may be turned out on to the palm of the hand. The film side is then uppermost, and the plate is to be transferred in the same position to a tray, and covered with water. This is to soak the film and obtain an equal action of the developer ; or the solution of fresh pyro may be poured on to the plate without previous soaking, if the flow is uniform, and the formation of bubbles avoided. In the first case the water is run off and the pyro allowed to flow evenly over the plate. To protect the plate from prolonged exposure to the ruby light, a second tray may be inverted over it, or the developing tray covered with a piece of card-board. Gently rock the tray for a minute or so, then to a few drops of ammonia in a measuring glass add the pyro from the developing tray, and pour the mixture back again over the plate. After again gently rocking the tray for a few minutes, more ammonia is added by drops in the same way. If the exposure has been properly timed, — and the time necessary must be ascertained by trial for each preparation, — the image will gradually begin to appear, and the action must be allowed to continue until sufficient density has been obtained. To determine this requires some experience. It is generally recommended to take the plate out of the tray and hold it for a moment, film-side towards the operator, in front of the ruby light. Though the plate is not nearly so sensitive when the image has commenced to develop, and there is, therefore, not the same danger of fogging, a safer plan is to occasionally turn the plate film downwards in the tray, and when the image appears on the back the development will be found to be completed. With such a preparation as tubercle bacilli in sputum it is not easy to trace the gradual formation of the image, and hence the advantage of commencing with a well-marked object such as the blow- fly's tongue. It is then easy to watch the gradual progress of the image. The bright parts or high-lights appear first, then gradually the half-tones, or less brightly-lighted parts, and lastly every shade except the deepest shadows is represented. When, however, all action seems to have ceased, we must still wait until we have judged, in the manner already described, that the density is sufficient. This being determined, we pour off the developing solution and thoroughly wash the plate with water. It is then ready to be placed in dish No. 1, containing " hypo," and here it must be left for some minutes after all appearance of creaminess has disappeared from the back. White light may now be admitted, the plate removed from the hypo and thoroughly washed under the tap, PHOTOGRAPHY OF BACTERIA. 167 and then placed in dish No. 2. When another plate is ready to take its place, transfer it to dish No. 3, and then to No. 4, and, after a good final washing under the tap, place it upon a rack to dry. If there is any tendency for the film to detach itself from the plate, " to frill," the alum bath must be used before fixing, as well as after. Frilling or blistering may be due to an error in manufacture, and. is liable to occur in hot weather, or when using a developer too strong in ammonia. If it occurs during washing or fixing, the alum bath must be employed before the hypo. Fogging, or the appear- ance of a veil over the plate, may arise from error in the manu- facture, from admission of extraneous light, from over-exposure, or from prolonged exposure to the ruby light during development. Care must be taken that the camera and dark-room are light-tight. Crystallisation, or powdery deposit, upon the negative when dry, is due to insufficient washing out of the hyposulphite of soda. Thin- ness of the image, or want of density, may be due to insufficient development, too weak a developer, or too short or too long an exposure. Too great density results from too long immersion in the developer. Spots may sometimes occur upon the negatives. They may be caused by dust upon the plate or by air bubbles in the developer. In the text-books of photography full accounts of failures will be found, their causes and prevention ; but it will be advantageous when these difficulties are encountered to take the negatives to a skilled photographer and get advice upon them. It is necessary to persevere, and not be disheartened if several negatives have to be made of a preparation before a successful result is obtained. It may here be remarked that the beginner will far more rapidly learn the technique if he avail himself of a practical demon- stration from a photographer. When he has learnt to obtain suc- cessful negatives, if he prefer silver prints, and time is an object, it will be found to be true economy to get the printing and mounting done by a professional photographer. The credit of a successful photograph of bacteria is due to the bacteriologist who prepares the microscopical specimen and obtains the negative. Determination of the Amplification. — The amplification varies not only with the objective employed, but with the distance of the focussing screen from the object. In order to ascertain the amplification afforded by a certain objective at a certain distance, a photograph should be taken, under the same conditions, of the lines of a micrometer slide. It is easy then to calculate the amplification 168 BACTERIOLOGY. obtained in the micro-photograph ; supposing, for example, in the micro- photograph the lines which are i^Vrr "1C^ apart are delineated 1 inch apart, the magnifying power must be 1,000 diameters. Moreover, having thus ascertained the amplification, we can accurately compute from the photograph the size of the objects taken. Value of Photographs. — It is not necessary to compare the relative merits of diagrams and photographs. Diagrams which do not purport to be accurate representations, but are intentionally the means for simplifying instruction, will always be valuable, even if we have the original preparations under the microscope before us. We must consider the relative merits of photographs and of drawings which purport to be exact representations of what is seen under the microscope. Thus in the case of micro-organisms, when their biological characters are studied under low powers of the microscope, photographs are preferable because they give a more faithful re- presentation. At the same time, apart from this comparative value, we must not lose sight of the actual value of photography in placing within the reach of the student or investigator, who may not be a draughtsman, a most valuable means for illustrating all kinds of preparations. For double-stained or triple -stained tissue pre- parations an accurately coloured drawing leaves CULTIVATION OF little to be desired ; but if we reproduce the same BACILLUS AN- \yy a woocl engraving, and so lose the advantage of the coloured picture, which is instructive in indicating the method of staining, a photograph will, in many cases, be far more satisfactory. When we have to deal with the growth of bacteria en masse, as in test-tube and plate- cultivations, with colonies as seen under a low power of the microscope, and with impression-prepara- tions both under low and high powers, unless the bacteriologist is a most accomplished draughtsman as well as an accurate and reliable observer, photography undoubtedly affords the best mode of illustration. The apparatus being ready and at hand, a negative can be produced in a few minutes of a preparation which, from the amount of detail it contains, would take perhaps several hours to draw and colour. From that negative any number of facsimiles can FIG. 82. — PHOTO- GRAPH OF PHOTOGRAPHY OF BACTERIA. 169 be obtained, whereas an original drawing, even in the best hands, if cut on wood or lithographed, is almost certain to fall short of being an exact copy. With regard to individual bacteria, the result is more satisfactory in many cases than a drawing ; for there is the advantage of being absolutely certain that any particular structure, form, or shape which may be represented is actually what exists, and not what may have been evolved by unconscious bias in the mind of the observer. Many illustrations might be given of this. Thus Lewis, who was a most conscientious observer, published an account of organisms in the blood of rats in India, and illustrated it with a wood engraving and with micro-photographs. The identity of the organisms which were found in the common brown rat of this country was established much more readily from • these photographs than from the wood engraving or the description in the letterpress. A micro-organism, even under the highest powers, appears as so minute an object that to represent it in a drawing requires a very delicate touch, and it is only too easy to make a picture which gives an erroneous impression to those who have not seen the original. If, on the other hand, to represent the object more clearly we draw an enlarged picture, we can only do so by repre- senting what we think the object would be like if it could be amplified to the size represented. In such cases a photographic enlargement is certainly more valuable. Photography enables us also to record rapid changes, and it is possible that as the art advances we may find that the film is more sensitive than the human retina, and brings out details in bacteria which would be otherwise unseen. Photographs can be readily transmitted by post, and when we can neither make a great number of preparations to illustrate !-orne object, nor perhaps be able to go to the expense of having a drawing reproduced, this method will be of value in enabling others to benefit by our observations. The author is convinced that if the employment of photography is encouraged in bacteriological and other research laboratories for depicting microscopical preparations and cultivations of bacteria, the results of increasing experience and practice will lead to its being made more general use of as a faithful and graphic method, valuable alike for class demonstrations and for illustrating publications. PAET II. ETIOLOGY AND PREVENTION OF INFECTIVE DISEASES. 171 CHAPTER XIII. SUPPURATION, PYAEMIA, SEPTICAEMIA, ERYSIPELAS. ABSCESS. WHEN inflammation is followed by an accumulation of leucocytes and of plasma which does not coagulate, the result is a white or creamy liquid called pus, and when the surrounding tissues are involved so that a cavity develops containing pus, we have what is termed an abscess. The almost constant association of bacteria with the production of pus has created a belief that they are the direct cause of suppuration. Ogston found micrococci present in all acute abscesses, and concluded that' acute inflammation was invariably due to their presence. The fact that inflammation occurs more frequently in the external tissues of the body is accordingly explained by the ready entrance of bacteria which are in the air ; and suppuration following pericarditis, pleurisy, and other conditions in which air is excluded is attributed to the presence of pyogenic cocci, which have gained access by the blood stream. There is no pyogenic organism constantly present, but several different .species of bacteria have been isolated from pus and carefully studied, and the antiseptic treatment is based upon the principle of excluding bacteria in surgical operations, and destroying any which may have previously obtained access to wounds and broken surfaces. Inflamed tissue and pus form a most suitable medium for the growth of bacteria, which in some cases are unquestionably only accidental epiphytes. In tuberculosis, actinomycosis, and glanders, pus formation may take place without the presence of pyogenic cocci ; and it is generally believed that chemical irritants, such as croton oil, turpentine, iodine, cadaverin, and tuberculin, will excite the formation of pus in the absence of bacteria, although Klemperer, after a number of very careful experiments, maintains that no genuine pus will be produced if the chemical irritants are first carefully sterilised. 173 174 INFECTIVE DISEASES. The bacteria which have been isolated from pus include : — Staphylococcus pyogenes aureus, albus, and citreus, Staphylo- coccus cereus flavus and albus, Streptococcus pyogenes, Micrococcus pyogenes tenuis, Micrococcus pneumonia crouposse, Bacillus pyo- cyaneus, Bacillus pyogenes foetidus, Micrococcus tetragenus, Bacillus intracellularis meningitidis, Gonococcus, Bacillus septicus vesicse, Urobacillus liquefaciens septicus, Bacillus typhosus, Bacillus coli communis, Bacillus anthracis, Bacillus tuberculosis, Bacillus mallei, and Actinomyces. FIG. 83. — SUPPURATION OF SUBCUTANEOUS TISSUE. d, Leucocyte containing micrococci ; d\ leucocyte with pale nucleus showing necrosis ; c, fixed connective tissue cells, much enlarged, containing several nuclei, of which some (n') are pale and necrotic ; numerous cocci, diplococci, and short chains. (CoRNiL and RANVIER.) Some idea of the distribution of the bacteria most commonly occurring in pus may be gathered from the records made by Passet and by Karlinski. Passet examined acute abscesses, and found Staphylococcus pyogenes aureus and albus in 11 cases, Staphylococcus pyogenes albus alone in 4, Staphylococcus pyogenes albus and citreus in 2, Streptococcus pyogenes alone in 8, Staphylococcus pyogenes albus and Streptococcus pyogenes in 1, and Staphylococcus pyogenes albus and citreus, and Streptococcus pyogenes in 1. SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 175 Karlinski tabulated his cases thus : — . A § .2 1 § i ^ 0 * 1 • " CO 3 £'£• I t || || iq II 5 3 11 jl | DISEASE. a "£>* ^g '>>%> 3 8> o w1 '§ 8 ll •<3 |l A fl ft CO II 1! fl CD | '§ PH "^ 00 PK fr 1 Mastitis 36 22 4 4 6 — Subcutaneous Abscess 30 10 2 8 6 2 2 — Phlegmon ..... 24 _ 24 — — Furuncle ..... 20 9 10 1 — — Bubo 17 8 1 1 7 — Subperiosteal Abscess 16 6 10 — Panaritium Cutaneum 16 7 9 — Panaritium Osseum . 10 7 3 — — Dental Abscess .... 10 1 4 1 3 1 — Horclcolviiii in i? 4* Abscess of the Middle Ear 1U 4 O 2 j _ 2 Carbuncle ..... 4 2 1 1 — 4 Osteomyelitis .... 3 2 — 1 — — — — — Total 200 82 7 55 45 6 3 2 4 PY^MIA AND SEPTICAEMIA. When pyogenic micrococci get access to the blood stream they may be carried into distant parts, and by multiplying produce meta- static abscesses in the lymphatic glands, bones, joints, and internal organs, a condition which is recognised as pyaemia. If there is a general invasion of the blood stream by micrococci, and absorption of their poisonous products, septicaemia results, and death may occur before the development of any secondary lesions. When septic micro-organisms multiply locally, and their chemical products are absorbed, or their products are separated from putrid material and injected into the circulation, the result may be called sapraymia. The blood in septicaemia contains living organisms, and is infective. The blood in sapraemia contains only the toxic chemical products, and is not infective. The one is septic infection and the other septic intoxication. Pyaemia may follow accidental wounds, surgical operations, parturition, acute suppuration of bones, scarlet fever, typhoid fever, and other diseases. To avoid pyaemia in surgery and midwifery, the greatest care must be taken to prevent micro-organisms from being conveyed by instruments, sponges, bandages, and by the hands of the surgeon or the obstetric physician. By the use of antiseptics and absolute cleanliness the chances of infection are reduced to a minimum. 176 INFECTIVE DISEASES. Rosenbach examined six cases of inetastatic pyaemia : Strepto- coccus pyogenes was found five times, partly in the blood and partly in the metastatic deposits, and twice in company with Staphylococcus pyogenes aureus. Baumgarten, also, found Streptococcus pyogenes in the internal organs in pysemic cases, and Eiselsberg found Streptococcus pyogenes in company with Staphylococcus pyogenes aureus in the blood of cases of septicaemia. Frankel isolated a streptococcus from puerperal fever, which he at first called Streptococcus puerperalis, but subsequently identified with Streptococcus pyogenes. These researches have been confirmed by others. Winkel obtained a pure cultivation of a streptococcus from the blood of the heart in a case of puerperal peritonitis. It produced erysipelatous redness when inoculated in the rabbit's ear, and in form arid in cultivation was similar to the streptococcus in erysipelas. Gushing also found Streptococcus pyogenes associated with puerperal infection. The cocci were found in endometritis diphtheritica as well as in secondary puerperal inflammation. These observations were still further confirmed by Baumgarten, and Bumm isolated the same organism in puerperal mastitis. DESCRIPTION OF BACTERIA IN Pus. A description may now be given of the cocci most frequently found. Staphylococcus pyogenes aureus and albus and Strepto- coccus pyogenes and Gonococcus are the most important of these. Staphylococcus pyogenes citreus, cereus albus and flavus, are pro- bably merely epiphytic. Micrococcus tetragenus, Micrococcus pyogenes tenuis, Bacillus pyogenes foetid us, Bacillus pyocyarieus, Bacillus coli communis, Bacillus septicns vesicse, Urobacillus lique- faciens septicus, arid Bacillus intracellularis meningitidis will be described fully in Part III. The description of Actinomyces, of Micrococcus pneumonise crouposse and of the bacilli of anthrax, tuberculosis, glanders, and typhoid fever, will be found in other chapters in Part II. Staphylococcus pyogenes aureus (Rosenbach). — Yellow coccus in pus (Ogston). Cocci singly, in pairs, very short chains, and irregular masses. Cultivated on nutrient agar-agar, an orange - yellow culture develops, looking like a streak made with oil paint. One variety grows on nutrient gelatine without liquefying it ; another produces rapid liquefaction, and the growth subsides as SUPPURATION, PYAEMIA, SEFOCLEMIA, ERYSIPELAS. 177 aii orange -yellow sediment. On potatoes and blood serum a similar orange-yellow culture grows luxuriantly. They may also form colourless growths in sub-cultures, and are then indistinguishable from Staphylococcus pyogenes albus. The cocci do not cause any septic odour in pus, nor does any gas develop. Albumin is con- verted by their action into peptones. They produce rapid ammoniacal fer- mentation in urine (Shattock). The micro-organisms injected into the pleura or knee of a rabbit produce, as a rule, a fatal result on the following day ; but if it survives longer, it eventu- ally dies of severe phlegmon. If injected into the knee of a dog, suppuration occurs, followed by disintegration of the joint. Injected into the peritoneal cavity of animals, they set up perito- nitis, and introduced into the jugular vein they produce septicaemia and death. When a small quantity of a cultivation is introduced into the jugular vein after previous fracture or contusion of the bones of the leg, the animal dies in about ten days, and abscesses are found in and around the bones, and in some cases in the lungs and kidneys, and the cocci are found in the blood and pus. Gar re caused sup- FIG. 84. — Pus WITH STAPHYLO- cocci, x 800 (FLUGGE). wound near his finger nail. Bockhart suffered from several pustules after vaccinating his arm with a pure-culture suspended in salt solu- himself a hypodermic FIG. 85.— SUBCUTANEOUS TISSUE OF A RABBIT 48 injectioii of a pure- HOURS AFTER AN INJECTION OF STAPHYLOCOCCI, X * «.»:>(> (IiAt-.M«jAKTKN). culture and produced an abscess. This micro- organism is practically ubiquitous. It has been cultivated from the skin and mucous membranes and secretions of healthy persons, a iul it occurs in the air, in soil, in dust, and in water, and in 12 178 INFECTIVE DISEASES. association with suppuration, pyaemia, puerperal fever, and acute osteomyelitis. Staphylococcus pyogenes albus (Rosenbach).— Cocci micro- scopically indistinguishable from the above. In cultivations also they resemble Staphylococcus pyogenes aureus, but the growth consists of opaque white masses. They, as a rule, liquefy nutrient gelatine very rapidly, and subside to the bottom as a white sediment ; more rarely they liquefy very slowly ; and a variety has also been described which does not produce any liquefaction. They are similar to the above-mentioned in their pathogenic action. Pure-cultivations of the organism were obtained from a case of acute suppuration of the knee-joint. Staphylococcus pyogenes citreus (Passet). — Cocci singly, in pairs, very short chains, and irregular masses. If cultivated on nutrient gelatine or nutrient agar-agar, a sulphur or lemon-yellow growth develops. When inoculated under the skin of mice, guinea- pigs, or rabbits, an abscess forms after a few days, from which a fresh cultivation of the micro-organism can be obtained. Staphylococcus cereus albus (Passet). — Cocci, morphologic- ally similar to the above, but distinguished by forming on nutrient gelatine a white, slightly shining layer, like drops of stearine or wax, with somewhat thickened, irregular edges. In the depth of gelatine they form a greyish- white, granular thread. In plate-cultiva- tions, on the first day, white points are observed, which spread themselves out on the surface to spots of 1 to 2 mm. When culti- vated on blood serum a greyish-white, slightly shining streak develops, and on potatoes the cocci form a layer which is similarly coloured. Staphylococcus cereus flavus (Passet). — Cocci which produce in nutrient jelly a growth which, at first white, becomes lemon- yellow, somewhat darker in colour than Staphylococcus pyogenes citreus. Microscopically Staphylococcus cereus flavus corresponds with Staphylococcus cereus albus. Inoculation experiments with both kinds give negative results. Streptococcus pyogenes (Rosenbach).~Cocci occurring singly, in masses, and in chains. The individual cocci are small spherical cells, with a special tendency after fission for the resulting elements to remain attached to each other, forming chains or rosaries. In cultures on solid media they often occur in the form of staphylococci, but in liquid cultures there may be a few, three or more elements, linked together ; or a great number, forming long chains which may be straight, serpentine, or twisted. DESCRIPTION OF PLATE IV. Streptococcus Pyogenes. FIG. 1.— -From a cover-glass preparation of pus from a pyaemic abscess. Stained with gentian-violet by the method of Gram, and contrast-stained with eosin. x 1200. Powell and Lealand's apochromatic -^ Horn. imm. E. P. 10. FIG. 2. — From cover-glass preparations of artificial cultivations of the strepto- coccus in broth and in milk at different stages of growth, x 1200. Powell and Lealand's apocbromatic ^ Horn. imm. E. P. 10. Jn these preparations there is a great diversity in size and form of the chains and their component elements. In the drawing examples are figured of the following: (a) Branched chains. (b) Simple chains composed of elements much smaller than the average size. v (c) Chains with spherical and spindle-shaped elements at irregular intervals. These are conspicuous by their size, and are sometimes terminal. (d e) Chains in which the elements are more or less uniform in size. (/) Complex chains with elements dividing both longitudinally and transversely, and varying considerably in size in different lengths of the sam.e chain. Plate IV xrx .* Figl. ../ Fig 2 STREPTOCOCCUS PYOGENES Vincent Bnolta, Day *Son,,Iil)i. SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 179 The individual elements composing the chains will be found to vary considerably in size : here and there in a preparation will be found a chain composed of excessively small cocci, in another part the elements will be all on a larger scale, and again in another part they will be peculiarly conspicuous on account of their si/o. So great is the diversity in the size of the cocci in some of the chains, that one might imagine that there was more than one kind of streptococcus present in a preparation, until on examining some of the longest chains it is observed that various sizes are repre- sented in different lengths of the same chain. Very characteristic appearances result from the fact that the cocci enlarge and divide both longitudinally and transversely ; and, indeed, the largest, for the most part, clearly show a division in two directions, resulting in the formation of tetrads. In addition to the forms resulting from the fission of the cocci, there are here and there in a chain, and sometimes terminally, larger elements, which are spherical, spindle-shaped, or in the form of a lemon. In the length of the chains, as in the size of the individual cocci, there is usually great diversity, In some cases they are composed of only a few, three or four cocci; in others eight, ten, or twenty. Here and there an exquisite rosary will extend in a straight line across the field of the microscope, or be twisted, curved, or serpentine ; in some preparations twisted or entangled strands are observed which are composed of several hundred elements. Such chains will be found to be much thicker in one part than another. Another char- acteristic appearance is produced by separation of the elements resulting from fission in the long direction of the chain, by which lateral twigs or branches are formed. Another character, which is very striking, may be seen when the individuals in a chain have become separated ; an unstained or faintly stained membrane may be found bridging across the interval. This will become still more visible in preparations contrast-stained with eosiu. In plate-cultivations the appearances of the colonies are not very striking. They appear to the naked eye after three or four days as extremely minute, greyish-white, translucent dots, which under the microscope have a slightly yellowish -brown colour. They are finely granular and well defined. They do not liquefy the gelatine, and after several weeks may not exceed the size of a pin's head. If the surface of nutrient gelatine solidified obliquely be traced over once or twice with a platinum needle bent at the extremity into a little hook charged with the cocci, a ribbon-shaped film develops in two or three days. This film is composed of minute, 180 INFECTIVE DISEASES. greyish-white, translucent dots or droplets, which can be more easily recognised with the aid of a pocket lens (Fig. 87). According to the number of organisms sown on the jelly, the dots or colonies may be completely isolated, or form a more or less continuous film. The film by reflected light has an iridescent appearance like mother-of-pearl, but has a bluish or bluish-grey tint by transmitted light, and with a pocket lens appears distinctly brownish. The gelatine is not liquefied, and even after several weeks the cultivation is limited to the inoculated area, and the individual colonies are, as a rule, not larger than pins' heads. In gelatine-cultivations of the same age, but kept in the incubator at 18° C., the colonies get irregular in form, especially at the margin of the film, and give the growth an arborescent, fringed, or serrated appearance. Cultivated on the oblique surface of nutrient agar-agar at 37° C. the growth is very similar, forming a film composed of minute white colonies like grains of sand ; but the film appears less transparent, is whiter, and the colonies have a greater tendency to get irregular in form. If inoculated with one tracing of the needle the growth is scanty, but tends to get thicker in the centre than towards the margins, which may have a terraced appearance. Inoculated in the depth of gelatine, there appears after a day or two at 18° C. a thread-like growth along the track of the inoculating needle. This delicate growth is found on examination with a pocket lens to consist of a linear series of extremely minute granules. In a few days more, the beads or granules become more marked, but even after weeks, the cultivation only appears like a string of minute, white, compact, globular masses or grains. In broth at 37° C. the cocci in twenty-four hours create a turbidity, and gradually develop beauti- ful chains varying in length according to the age of the cultivations. Even in forty-eight hours there may be chains of eight, ten, twenty, or a hundred elements. After a few days the growth settles down at the bottom of the tube in the form of a white deposit, while the supernatant liquid becomes again clear. Inoculated subcutaneously in the ear of rabbits, they produce in. two days an inflammatory thickening with erysipelatous redness, or sometimes suppuration. They may occur in vaccine lymph, as the result of con- tamination, and Pfeiffer suggested that before calf lymph is employed for vaccination it should be tested on a rabbit's ear. If in two days no rash has been produced, the possibility of the presence in the lymph of Streptococcus pyogenes or erysipelatis is excluded. SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 181 According to Fliigge and others, after subcutaneous inoculation of mice with a small quantity of a cultivation, there is no result in 80 per cent, of the animals experimented upon. Sometimes there is limited pus formation at the seat of inoculation, sometimes the animals die without any very striking pathological appearances. They occur in abscesses, pyaemia, and septicaemia, and are often found in diseases such as scarlet fever and typhoid, associated with septic complications. They have been isolated from air, soil, and water. The streptococcus found in erysipelas agrees in description, and is merely a variety of Streptococcus pyogenes. It has been definitely established by the researches of Frankel and Freudenberg, and later by those of the author, Raskin, Prudden, and Bayard Holmes, that Streptococcus pyogenes is frequently found in scarlet fever and diphtheria, and in other diseases associated with septic complica- tions. The author has isolated Streptococcus pyogenes from acute abscesses, from suppuration after surgical operations, from pyaemia, from pyaemia after scarlet fever, and from purulent peritonitis. Some of these cultures have been kept up for very long periods, extending over some years, so that opportunities occurred for a complete investigation into the life history of this micro-organism. Variations in the appearances of cultures have been observed when obtained from the same source. A number of cultures from pus were prepared 011 gelatine and agar, made according to the usual formula, but at different dates, and, therefore, varying slightly in composition and quality. Sub-cultures were also started in nutrient gelatine of precisely the same composition, but from primary cul- tures of the same micro-organism in different media — agar-agar, milk, and broth. The descriptions of the streptococcus hitherto published were then found to be inadequate. The different cultures and sub-cultures presented striking variations in the microscopical and macroscopical appearances. Some sub-cultures on gelatine, for exam- ple, exhibited a finely dotted appearance, others showed every variety in the size, and degree of opacity of the colonies (Fig. 89). Cultures in broth also, varied in appearance, owing to slight variation in the composition of the medium, to slight differences of temperature, an«l other conditions difficult to determine. The addition of glycerine to broth materially alters the appearance of the culture. It was con- clusively proved that minute differences arise from different conditions of the cultivating media. The author was led to study exhaustively the streptococcus of acute suppuration in bo vines. Primary cultures of Streptococcus pyogenes from man, and primary cultures from 182 INFECTIVE DISEASES. a case of purulent peritonitis in a cow, were carried through sub-cultures under exactly similar conditions. -Cultivations of the Streptococcus pyogenes bovis exhibited variations in microscopical and cultural characters which were even more marker! than in the case of the Streptococcus pyogenes hominis. By selecting certain cultures from both sources there was a striking similarity if not identity between them, but, when compared under exactly identical conditions, there was more difference in cultural characters between the Streptococcus pyogenes bovis and the Streptococcus pyogenes hominis than between the Streptococcus pyogenes hominis and the Streptococcus erysipelatis, and they may therefore be regarded as distinct varieties (Figs. 89, 90). Some of the diseases and conditions in which Streptococcus pyogenes has been found may be alluded to more in detail. Spreading Gangrene.— From, a case of spreading gangrene, which was identical with Ogston's erysipelatoid wound gangrene, and regarded by him as the most intense and dangerous form of erysipelas, Rosenbach obtained pure-cultivations of a streptococcus by incising the skin of the limb, and inoculating tubes from the turbid reddish fluid which escaped. That the streptococcus was identical with Streptococcus pyogenes was ascertained by comparison with a cultivation derived from pus, of the mode of growth, and of the effect on animals. Surgical Fever. — Eiselsberg proved the presence of a streptococcus in the blood of several cases of surgical fever in Billroth's clinic. The organism was identified by cultivation with Streptococcus pyogenes. Diphtheria. — In three cases of typical diphtheria, Loffler found a streptococcus. He isolated it by cultivation, found that it was similar in form, characters on cultivation, and effects after inoculation, to Fehleisen's streptococcus of erysipelas. Loffler was not inclined to regard them as identical, because Fehleisen never found his cocci in the blood-vessels. Fltigge named the organism Streptococcus articulorum, and states, that after subcutaneous inoculation or injection of a cultiva- tion in mice, a large proportion of the animals die, and in the sections of the spleen and other organs the streptococci are again seen. Baum- garten investigated the same subject, and decided that the streptococcus was identical with Streptococcus pyogenes. Small-pox. — Hlava has established the presence of Streptococcus pyogenes in the pustules of variola, and Gam' found streptococci in the internal organs in a case of variola haBmorrhagica. In a fatal case of variola complicated with pemphigus, Garre found a streptococcus in the pemphigus vesicles. Whether it was identical with Streptococcus erysipelatis Garre left an open question. Yellow Fever. — Babes observed the presence of streptococci in the vessels of the kidney and liver in yellow fever. Cultivation experiments are wanting. It was probably a case of secondary infection with Strepto- coccus pyogenes. SUPPURATION, PY.EMIA, SEFHC^MIA, ERYSIPELAS. 183 Bilious Fever, — Babes, in a case of fievre bilieuse typhoide, found masses of streptococci filling the vessels of the liver, kidney, and spleen. This was probably another instance of secondary infection with Strepto- coccus pyogenes. Measles. — From the blood and inflammatory post-products in measles, Babes isolated a streptococcus, which he describes as closely resembling the Streptococcus pyogenes. Ulwiitfrr l-liijocdi-dit/x. — Wyssokowitsch found cocci in the internal organs in ulcerative endocarditis, and produced the disease in animals, after injury to the valves, by injection of Streptococcus pyogenes and other organisms. Weichselbaum, by microscopical research and by cultivation experiments, proved the presence of Streptococcus pyogenes in acute verrucous endocarditis. Baumgarten confirmed this. He found FIG. 86.— ULCERATIVE ENDOCARDITIS : SECTION OF CARDIAC MUSCLE, x 700 (Kocn). Streptococcus pyogenes alone in one case and accompanied by Staphylo- coccus aureus in another. Broncho-pneumonia, — Thaon found a streptococcus in the lungs of children in fatal cases of broncho-pneumonia, complicating measles, diphtheria, and whooping cough. It was regarded as identical with the streptococcus isolated by Loftier from diphtheria. Frankel discovered a streptococcus in the lungs of a case of true croup complicated with broncho-pneumonia, and by cultivation established its identity with Streptococcus pyogenes. Anthrax. — Charrin found cocci in rabbits, examined some hours after death from anthrax. These, when isolated, produced death in rabbits from septicaemia, without suppuration. Chains composed of from fifteen to twenty elements were found in all the organs. This was probably another instance of Streptococcus pyogenes. SypJrili*. — Kassowitz and Hochsinger found the presence of a strepto- 184 INFECTIVE DISEASES. coccus in the tissues and internal organs, and especially in the blood- vessels, in fatal cases of congenital syphilis. These observers regarded their discovery as having an important bearing on the etiology of syphilis, but Kolisko pointed out that it was only the result of septic infection with presence of Streptococcus pyogenes, as had already been established in scarlet fever. Cerebro- spinal Meningitis. — From the meningeal exudation of a case of apparently idiopathic cerebro-meningitis, Banti found Streptococcus pyogenes and Staphylococcus aureus and albus. The cocci probably entered through an abscess of the jejunum. FlG. 87. — PURE-CULTURES OF STREPTOCOCCUS PvOGENKS. a, On the surface of nutrient gelatine ; I, in the depth of nutrient gelatine c, on the surface of nutrient agar. Blepharadenitis and Dna-ywyxfix. — Widmark isolated by cultivations Streptococcus pyogenes and other organisms from cases of blepharadenitis and phlegmonous dacryocystis. In phlegmonous dacryocystis Widmark found Streptococcus pyogenes almost exclusively. LeukcBmia. — Fliigge cultivated a streptococcus from necrotic patches in the spleen of a fatal case of leukaemia. Cultures corresponded very closely with Streptococcus pyogenes. Inoculation in the ears of rabbits produced similar results to Streptococcus pyogenes or erysipelatis. Fliigge calls it Streptococcus pyogenes malignus, but concludes that it is probably dentical with the streptococcus from pus. SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 185 ERYSIPELAS. Erysipelas is an acute inflammation of the skin, occurring in connection with wounds, when it is traumatic, and on surfaces apparently sound, when it is idiopathic, as in erysipelas of the face. It is highly contagious in surgical wards, and it gives rise to rapidly fatal puerperal fever in lying-in hospitals. In such t-ascs the virus is obviously conveyed from sick to healthy persons by direct contact, or by instruments and sponges, or by the hand of the surgeon, physician, or nurse, and possibly by the air. FIG. 88.— SECTION OF SKIN IN ERYSIPELAS. v,v, two lymphatic vessels containing leucocytes and m,in, streptococci ; t, connective tissue ; a, connective tissue and wandering cells, x 600 (CoRNiL and RANVIER). Streptococcus of Erysipelas. — In 1882, Fehleiseii isolated a streptococcus in erysipelas, described the appearances on cultivation, and maintained that it could be distinguished from the streptococcus of suppuration. Rosenbach agreed that the two micro-organisms could be distinguished by parallel experiments, and named the one Streptococcus pyogenes and the other Streptococcus erysipelatis. (Fehleisen). Rosenbach asserted that the colonies of the latter were more opaque and whiter than those of Streptococcus pyogenes and the growth more marked in the depth of nutrient gelatine, while microscopically the chains were better marked and larger, and the individual cocci larger than in Streptococcus pyogenes. Others who investigated this subject could not distinguish them with certainty, either by their morphological or cultural characters or effects on inoculation. Passet found that inoculation of Strepto- coccus pyogenes induced a condition very similar to that produced by inoculation of. Streptococcus erysipelatis. Hoffa arid Hajek described minute differences, but Biondi and Eiselsberg failed to confirm these. 186 INFECTIVE DISEASES. Baumgarten failed to prove any essential difference. Mitchell Prudden found that Streptococcus pyogenes injected into the sub- cutaneous . tissue of the ears of rabbits, produced in one no effect ; in four, slight transient redness ; in five, local redness followed by abscess ; in twelve, well-marked erysipelatous redness, followed by complete resolution in seven, abscess in three, and death in two. Passet, Biondi, Eiselsberg, Baumgarten, and Mitchell Prudden concluded that, in their morphological, biological, and pathogenic characters, so far as animals are concerned, the two organisms are practically identical. The author investigated the morphology and cultural characters of the Streptococcus erysipelatis, which he had isolated from a typical case. This result cleared up the conflicting statements which had been made by different observers. By carrying out absolutely parallel experiments, the Streptococcus pyogenes and Streptococcus erysipelatis were unquestionably distinguishable, as Fehleisen and Rosenbach had asserted. In both cases, however, inoculation of a trace of a culture from a solid medium produced only transient redness. Injection hypodermically of a broth-culture produced in both cases a spreading erysipelatous redness, followed by suppuration. It was found that primary cultures of the two micro-organisms, cultivated under precisely the same conditions, differed in the size and character of their chains, in the size of the individual elements, in the greater opacity of the colonies of Strepto- coccus erysipelatis, in a greater tendency to confluence, and in a more rapid growth. The author found that the difference was most marked in broth-cultures. Abundant flocculi were formed by Streptococcus pyogenes ; a powdery deposit with special tendency to form a granular adhesive film at the bottom of the culture flask, in the case of the streptococcus of erysipelas. Lastly, they differed in their power of resisting germicides. Fehleisen inoculated patients in hospital suffering with malignant growths, and produced a typical erysipelas with sub-cultures after an incubation of from sixteen to twenty hours. The disease was marked by rigors, fever, and general disturbance. Patients who had recently suffered from erysipelas had an immunity. Emmerich succeeded in proving the presence of streptococci in the air of a hospital where erysipelas had broken out. These cocci in their form, their characters on cultivation, and their inoculation results, were identified with the Streptococcus erysipelatis. It is riot therefore exclusively parasitic. Streptococci identical or agreeing very closely in their description SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 187 \vitli Streptococcus pyogenes. have been found in cattle plague, foot and mouth disease, strangles, contagious mammitis in cows, and progressive tissue necrosis in mice, and they will be referred to fully in subsequent chapters. EXAMINATION AND CULTIVATION OF STREPTOCOCCI. Cover-glass preparations can be stained with the watery solutions of the aniline dyes. In some cases very beautiful preparations can be obtained by using Neelsen's solution, and removing excess of b. c. . d. FIG. 89. — STREPTOCOCCUS PYOGENES HOMINIS. Pure-cultures on nutrient gelatine. «, Sub-culture from agar. c, Sub-culture from milk. b, Sub-culture from broth. d, Sub-culture from milk. stain by rinsing in alcohol. To examine pus, milk, or broth, take nn ordinary platinum needle bent at the extremity into a booklet. Dip it into the liquid to be examined, and spread it on a cover- idass into as thin a film as possible; the preparation is treated in the ordinary way, that is 'to say, the film is allowed to dry, and the cover is taken up with forceps, and passed three tinio through the flame with its prepared side uppermost. Gram's Method with Eosin. — In this way the streptococci art- stained blue, and stand out in marked contrast to the rest of the preparation. Use freshly prepared solution. Float tbe cover-glasses 188 INFECTIVE DISEASES. on the solution for ten minutes to half an hour, then trai.sfer them to iodine-potassic-iodide solution, until they assume the colour of a tea leaf ; then immerse them in alcohol until they are decolorised ; dip them in an alcoholic solution of eosin for a few moments, and then transfer them to clove oil to clarify the film ; to remove the clove oil gently press the cover between two layers of clean filter paper, then mount in xylol balsam. A good method for cultivating streptococci is to employ a steril- ised looped platinum wire, and to spread a droplet, for example, of pus or blood, over the surface of nutrient agar-agar solidified obliquely. a- '>• c. FIG. 90.— STREPTOCOCCUS PYOGENES Bovis. Pure-cultures on nutrient gelatine. a, Sub-culture from agar. b, Sub-culture from broth. c, Sub-culture from milk. d, Sub-culture from milk. The tubes are then placed in the incubator at 37° C. ; the strepto- cocci will appear in the course of two or three days in the form of minute dotted colonies. If present alone, and in considerable quantities, the inoculated surface will exhibit a pure cultivation consisting of a number of such colonies, whilst a flocculent mass is observed in the liquid which collects at the bottom of agar-agar tubes ; this flocculent mass will be found to be composed of chains. From such a tube inoculate a number of the small flasks employed in Pasteur's laboratory for cultivations in liquids. In this way a number of pure- cultivations in milk and broth are established, which can le readily examined from time to time. From a pure- SUPPURATION, PYAEMIA, SEPTICAEMIA, ERYSIPELAS. 189 cultivation in broth or agar-agar tubes of nutrient gelatine can be inoculated. Cover-glass-preparations from the growths on solid media can be made in the usual way, and stained with either a watery solution of fuchsine or gentian violet : but to stain prepa- rations made from milk or broth, or from the liquid in agar-agar tubes, use the method of Gram ; the stain will then be removed, except from the streptococci, .and very beautiful preparations result. GONORRHCEA. Gonorrhoea is the result of a catarrhal inflammation of the mucous membrane of the urethra, vagina, or conjunctiva caused by a characteristic pyogenic organism discovered by Neisser in 1879. Gonococcus of Neisser. — Cocci, usually in pairs 1-6 p in length, -8 ^ in width, and tetrads, with those surfaces of the com- ponent elements which are in contact, flattened. The elements are more or less kidney-shaped, and are separated by a clear unstained interval. They are found free in the pus and also in the interior of the pus cells. They stain with the aniline dyes, but are decolorised by Gram's solution. They do not grow on the ordinary media, such as gelatine, agar, and potato, in marked contrast to the common pyogenic cocci ; but Bumm succeeded in obtaining a cultivation by using human blood serum, which was procured for the purpose from the placenta. They give rise to a very delicate growth in the form of an almost invisible film, with a moist appearance, which attains its full development in a few days. Steinschneider used human blood serum and aufar incubated at 35° 0. Krall recommended either agar with grape-sugar and blood serum, or the same mixture with the addition of 5 per cent, glycerine. Others have employed nutrient agar with the surface moistened with sterilised human blood. More recently Keifer has been successful with a medium which is prepared in the following way : a si-it ic- fluid is filtered arid sterilised by Tyndall's process, to this is add«'d an equal quantity of the following mixture, agar 3'5, peptone 5, glycerine 2, salt '5 (per cent.). The ascitic agar is solidified in a Petri's dish, and the culture incubated at 36° C. They have also been cultivated in albumin from plovers' eggs, and in the fluid obtained from a case of synovitis of the knee joint. Inoculation of rabbits, dogs, horses, and monkeys, has been invariably unsuccessful, but sub-cult -nros produce the disease in the health urethra. OF TBK UNIVERSITY 190 INFECTIVE DISEASES. The cocci are found in pus from the urethra and other mucous membranes affected by the disease. They have also been found in urethral and inguinal abscesses in association with Staphylococcus pyogenes aureus. METHOD OF STAINING. Cover-glass preparations are made in the usual way, and double stained with Loffler's methylene blue, and eosiii. Schiitz recommends floating the cover-glasses for five or ten minutes in a saturated solution of methylene blue in 5 per cent, solution of carbolic acid. They are washed in water, rinsed in very weak acetic acid, and again washed in water. Safranin may be used as a contrast stain. FIG. 91.— GONOCOCCUS x 800 (BUMM). a, free cocci ; b, cocci in pus cells ; c, epithelial cell containing cocci. EGYPTIAN OPHTHALMIA. There are two forms of ophthalmia in Egypt, one associated with Gonococcus and the other with a bacillus closely resembling the bacillus of mouse-septicaemia, but there are minute differences. Bacillus of Ophthalmia (Koch and Kartulis). Minute rods which do not grow on gelatine but readily on blood serum and nutrient agar, forming a plainly visible, whitish-grey shining growth. Animals are insusceptible, but cultures produced the disease in the human conjunctiva in two out of six cases. CHAPTER XIV. ANTHRAX. ANTHRAX is a very fatal malady, and most irregular in its be- haviour. At one time it attacks only one or two animals, and at another time it will destroy nearly all the stock on a farm. Farmers formerly regarded the disease as non- communicable, and possibly the result of excessive or improper feeding, or faulty sani- tation, or of climatic conditions over which no control could be exercised. It is obvious that so long as the disease was regarded as the result of unknown conditions, no explanation could be given of its recurrence from time to time, or of certain animals contracting the disease and others not, and no measures of any use could be suggested to cope with an outbreak. Anthrax has always been more prevalent on the Continent than in England, and this to some extent accounts for the fact that it has received greater attention abroad. In France, Germany, Hungary, Russia, and in India and Persia, anthrax at times produces wide- spread losses. In Siberia it is still known on this account as the Siberian Plague. On the Continent there are certain localities known as anthrax districts on account of their reputation for anthrax — for example, in the Upper Bavarian Alps in Germany and in Auvergne in France. In 1849, Pollender happened to examine the blood of a cow after death from anthrax, and discovered peculiar rod-like bodies among the blood cells. The same observation was made independ- ently by Brauell and Davairie about the same time, but the greatest importance must be attached to the publication of Davaine's further researches in 1863. Many ridiculed the discovery of bacilli, and stoutly maintained that they were only blood crystals or accidental structures of no importance. For many years very little progress was made, and the statements of other observers who were able to verify and add to Pollender's and Davaine's discoveries, were still received with scepticism. 191 192 INFECTIVE DISEASES. Within the last few years a great change of opinion has taken place. Bacteriologists have investigated the whole subject, so that at the present day we know exactly the cause of anthrax. Bacillus anthracis (Bacteridie du charbon, Bacillus of splenic fever, Wool-sorters' disease, or malignant pustule). — Rods 5 to 20 ^ long and 1 to 1'25 p broad, and threads; spore-formation present. As a thorough knowledge of the life-history of this bacillus is of the greatest importance, the various steps to be followed in a practical study of it will be successively treated in detail. Its morphological * vf.m ™ /a£8 FIG. 92. — BACILLUS AXTHRACIS, x 1200. Blood corpuscles and bacilli unstained ; from an inoculated mouse (FRANKEL and PFEIFFER). and biological characteristics have been very completely worked out, and it serves as an excellent subject for gaining an acquaintance with most of the methods employed in studying micro-organisms. A mouse inoculated with the bacillus or its spores will die in from twenty-four to forty-eight hours, or more rarely in from forty-eight to about sixty hours. Examination after Death. — The spleen is found to be considerably enlarged, and may be removed, and examined by making cover-glass preparations, inoculations in nutrient media, and subsequently sections. Cover-glass Preparations. — In cover-glass preparations of the blood of the spleen the bacilli are found in enormous numbers. Preparations should also be made with blood from the heart and with the exudation from the lungs and other organs ; it will be DESCRIPTION OF PLATE Y. Bacillus Anthracis. FIG. 1. — From a cover-glass preparation of blood from the spleen of a guinea- pig inoculated with blood from a sow. x 1200. Powell and Lealand's apochromatic T\ Horn. imm. E. P. 10. FIG. 2. — From a section of a kidney of a mouse. Under a low power the preparation has exactly the appearance of an injected specimen. Under higher amplification the bacilli are seen to have threaded their way along the capillaries between the tubules, and to have collected in masses in the glomeruli. Stained with Gram's method (gentian-violet), and eosin. x 500. FIG. 3. — Bacillus anthracis and Micrococcus tetragenus. From a section from the lungs of a mouse which had been inoculated with anthrax three days after inoculation with Micrococcus tetragenus. A double or mixed infection resulted. Anthrax-bacilli occurred in vast numbers, completely filling the small vessels and capillaries, and in addition there were great numbers of tetrads. Stained by Gram's method (gentian-violet), and with eosin. x 500. Plate V. -^ \ y/s js» * I'xC^ / Fig Fig 2. Fig 3. BACILLUS ANTHRACIS ANTHRAX. 193 noted that in these the bacilli are present in very small numbers, or altogether absent. The bacilli should be examined both unstained and stained. The rods are straight or sometimes curved ; rigid and motionless. They can be stained with a watery solution of any of the aniline dyes, and are then seen to be composed of segments with their extremities truncated at right angles ; between the segments a clear linear space exists, which gives them a characteristic appear- ance (Plate V., Fig. 1). By double staining, with Gram's method and eosin, the rods are seen to consist of a membrane or hyaline sheath with protoplasmic contents. Drop-cultures. — A little of the blood from the spleen or heart may be employed to inoculate sterilised broth or blood serunu Several of these cultures should be prepared, and some of them placed in the incubator, and examined at intervals of a few hours. It will be observed that the rods grow into long homogeneous fila- ments, which are twisted up in strands, and partly untwisted in long and graceful curves. The filaments begin to swell, become faintly granular, and bright, oval spores develop (Plate 1). The cultures in the incubator develop rapidly. A temperature of 30° to 37° C. is the most favourable for spore-formation. The spores are eventually set free, and by making a fresh cultivation, or by injecting them into a mouse or guinea-pig, they germinate again into the characteristic bacilli, which in their turn grow into filaments and spores. When the spore germinates it swells, the envelope becomes jelly- like, and gives way at one or other pole, and the contents escape and grow into a rod. Test-tube Cultivations in Nutrient Gelatine. — Typically characteristic appearances are obtained by inoculating a 5 to 8 per cent, nutrient gelatine. A whitish line develops in the track of the inocu- lating needle, and from it fine filaments spread out in the surrounding medium (Fig. 93). The fila- FIG. 93.— PURE CUL- ments are more easily observed with a magnifying TIVATION OF BA- glass. In a more solid nutrient gelatine the growth appears only as a thick white thread. As lique- LATINE. faction of the gelatine progresses, these appearances gradually alter, and the growth subsides to the bottom of the tube as a white flocculent mass. In exhausted culture- media, and sometimes in the blood, filaments are seen in a state of degeneration. 13 194 INFECTIVE DISEASES. This has also been observed in sections of the internal organs of a rabbit which had been inoculated with the anthrax bacillus and had. died of septicaemia the following morning. Test-tube Cultivations in Nutrient Agar-agar. — Cultivated upon a FIG. 94.— COLONIES OF BACILLUS ANTHRACIS, x 80 (FLUGGE). a, after 24 hours ; b, after 48 hours. sloping surface of nutrient agar-agar a viscous snow-white layer is developed, but without access of air no cultivation can be obtained, the bacilli being aerobic. This can be demonstrated by completely embedding a piece of lung or spleen pulp containing bacilli, in nutrient agar-agar (p. 22). Potato - cultivations. — In about thirty -six to forty-eight hours a creamy- white or very faintly yellowish layer forms over the inoculated surface, usually with a translucent edge, and sometimes a strong, penetrating odour of sour milk. Plate-cultivations. — From the spleen or blood of the heart cultivations may be made in nutrient gelatine on plates. The colonies develop in about two days, according to the temperature of the room. They appear to the naked eye as little white spots FIG. 95.— IMPRESSION-PREPARA TION OF A COLONY, X 70. ANTHRAX. 195 or specks, which, on examination with a low power of the microscope and small diaphragm, exhibit two distinct forms. One form, on careful focussing, has the appearance of a little compact ball of FIG. 96.— MARGIN OP A COLONY, x 250 twisted threads ; in the other, liquefaction of the gelatine has commenced, and the threads spread out like locks or plaits of hair in the neighbouring gelatine. These appearances are perfectly characteristic (Figs. 94, 96). Cover-glass Impressions. — The plate- cultivations should be also examined as soon as the colonies appear, by making cover-glass im- pressions (Fig. 95). The filaments, examined with a high power, will be seen to consist of a number of rods or segments which are perfectly regular in form. On the other hand, filaments from a tube-cultiva- tion in a solid medium will often be found to be composed, not only of rods, but here and there of the so- called involution-forms (Fig. 97). FIG. 97.— FILAMENTS WITH OVAL AND „ , , . i IRREGULAR ELEMENTS, X 800. From cultures m gelatine and glycerine agar, very striking preparations are sometimes 'obtained, with numerous large spherical and lemon-shaped elements. In a 196 INFECTIVE DISEASES. cover-glass preparation from a potato-culture the individual segments will be found to have a great tendency to be isolated one from the other, and there is copious spore-formation. Preservation of Spores. — .Spores may be preserved simply by allow- ing anthrax blood to dry and then sealing it in a tube. The spores from a potato-cultivation are treated as follows : — The inoculated surface bearing the creamy cultivation is sliced off in a thin layer, and is mashed up with distilled water in a glass capsule. Sterilised silk-thread is cut up into lengths of about a quarter of an inch, and allowed to soak in the paste for some hours, under a bell-glass. The threads are then picked out with a pair of forceps, and laid upon a sterilised glass plate, covered with a bell-glass, and allowed to dry. From the plate, when perfectly dry, they are transferred to a small test-tube, which can be plugged with cotton-wool, or sealed in the Bunsen burner. Examination of the Tissues. — The organs should be hardened in absolute alcohol, and sections prepared and stained by the ordinary methods. The method of Gram is the most instructive, and eosin a very satisfactory contrast stain. The capillaries in the lungs, liver, kidney, spleen, skin, mucous membrane, etc., will be found to contain bacilli. In some cases the bacilli are so numerous that a section under a low power has the appearance of an injected specimen. Inoculation of Animals. — A thread containing spores, a drop of blood from an infected animal, or a minute portion of a cultivation, introduced under the skin of a mouse or guinea-pig, causes a fatal result, as a rule, in from twenty-four to forty-eight hours. Sheep fed upon potatoes which have been the medium for cultivating the bacillus, die in a few clays. Goats, hedgehogs, sparrows, cows, horses, swine, and dogs are all susceptible. Rats are infected with diffi- culty. Frogs and fish have been rendered susceptible by raising the temperature of the water in which they lived. Cats, white rats, and Algerian sheep have an immunity from the disease. Attenuation of the Virus. — Toussaint attenuated cultures by exposing them for ten minutes to 55° 0. Pasteur obtained a similar- result by resorting to lower degrees of temperature ; and Koch, Gaffky, and Loffler concluded from their experiments, that from 42° to 43° C. the bacillus was most easily deprived of its poisonous properties. By cultivating the bacillus in neutralised broth at 42° to 43° C. for about twenty days, the infecting power is weakened, and animals inoculated with it (premier vaccin) are protected against the disease. To obtain a still more perfect immunity, they are ANTHRAX. 197 inoculated a second time with material (deuxieme vaccin) which has been less weakened. The animals are then protected against the most virulent anthrax, but only for a time. From a weakened culture, according to Klein, new cultures of virulent bacilli can be started, and a culture that can be used as a vaccine for sheep kills a guinea-pig, and then yields bacilli that are fatal to sheep. The virulence of the bacillus is also altered by passing the bacillus through different species of animals. The bacillus of sheep or cattle is fatal when re-inoculated into sheep or cattle; but if inoculated in mice, the bacilli then obtained lose their virulence for sheep or cattle, only a transitory illness results, and the animals are protected for a time against virulent anthrax. Exposure to a temperature of 55° C., or treatment with *5 to 1 per cent, carbolic acid, deprives the bacilli of their virulence. Chauveau obtained a similar result by cultivating the bacillus at 38° or 39° C. under a pressure of eight atmospheres. The possibility of mitigating the virus depends upon the species of animal ; rodents cannot be rendered immune by any known anthrax vaccine. The nature of the toxic products has been described in a previous chapter (p. 42). METHODS OF STAINING THE BACILLUS ANTHRACIS. Cover-glass preparations of blood, etc., can be stained with a watery solution of any of the aniline dyes, or with Neelsen's solution and subsequent treatment with alcohol (p. 87). The preparations may be dried and mounted permanently in Canada balsam, but the typical appearances are best observed in freshly stained specimens examined in water. The sheath and protoplasmic contents can be demonstrated in cover-glass preparations from the blood or spleen which have been stained with eosin after the method of Gram. Spores must be stained by the ^ special methods already described. /^^ C^ The most satisfactory preparations are obtained by double-staining with FIG. 98.— SPORES OF BACILLUS Ziehl-Neelsen solution and methy- ANTHRACIS STAINED WITH , , /T^. ~v GENTIAN VIOLET, x 1500. lene blue (Fig. / ). Tissue sections are best stained by the method of Gram, and after-stained with eosin, picrocarminate of ammonia, or picro-lithium- carmine. 198 INFECTIVE DISEASES. ORIGIN AND MODE OF SPREAD. As every outbreak of anthrax is the result of the introduction into the system of the bacilli, the question naturally arises, how are they introduced on the farm ? Where do they come from ? and what are the channels of infection ? The spores of the bacilli may get into the soil, and may remain there in a dormant state for many years. The spores were believed by Pasteur to be taken up by earth-worms, carried to the surface and deposited in their castings. Animals grazing are thus liable to be infected; but Koch's experiments tended to disprove this theory. Anthrax has been known to break out among cattle grazing on a field where several years previously some Russian hides from infected animals had been buried. By some means or other the spores may contaminate the grass, and hay imported from an anthrax district may start the disease on a farm on which it had never been known to occur. The spores may in a similar way be introduced with blood manure and bone manure, and with refuse used as manure. The skin, hair, wool, hoofs, and horns of infected animals, if soiled with blood, are contaminated by the bacillus. Another way in which the disease can be communicated may be illustrated by the transmission of the disease to man. Those who handle carcasses, wool or hides of infected animals are liable to contract the disease. Slight scratches, cuts, bites, and pimples, may readily be inoculated with the bacilli or their spores. Veterinary surgeons, butchers, herdsmen, cattle drovers — in fact, all those whose occupation leads them to cut open or skin cattle, sheep, or horses, or to handle hides and wool — are liable to fall victims to this disease. In one case which was brought to the author's notice, a veteri- nary surgeon had been called to see a bullock which had died suddenly in a meadow. A post-mortem examination was made, and the veterinary surgeon wiped his hands, which were soiled with blood, on some rough grass, and then washed them in a stream. The sedgy grass made some small cuts on his fingers, and the result was that he was simultaneously inoculated with the blood of the bullock. Local anthrax followed, two of his fingers were amputated, and he fortunately recovered. In another case a butcher dressed the carcass of a beast which had died suddenly, and while doing so scratched a pimple on his neck. An anthrax pustule developed, and after a very serious illness he also recovered ; but in many cases the attack is fatal. " Wool -sorters' disease" is ANTHRAX. 199 anthrax of the lungs. Bales of foreign wool contain not only wool from living sheep, but wool which has been clipped from skins of dead sheep. If any of the sheep died from anthrax the wool is sure to be contaminated with blood containing the bacilli, and then wool-sorters engaged in picking the wool readily inoculate them- selves through a scratch or pimple, or by inhaling the spores. In many cases Wool-sorters' disease is fatal. A farm may become extensively infected by the living animal. Blood containing the bacilli may be discharged from the mouth and nostrils, or be passed with the contents of the intestinal canal and bladder. The droppings contaminate the pasture or byre, and spore formation, especially in warm weather, quickly takes place. From this cause the disease may not only be conveyed to healthy cattle grazing with infected animals, but fresh cases may occur, year after year, on the same far in, and if hay is cut and sold off the farm, other cattle at a distance are similarly infected. If the flooring of cattle sheds is once soiled by infected animals it is easy to account for those otherwise mysterious outbreaks which occur when the cattle are taken in for the winter. Another source of danger arises when blood from a diseased animal is washed into brooks or streams, for thus the disease may be carried to farms in which it was previously unknown. PREVENTIVE MEASURES. Early recognition and prompt action are essential to prevent the spread of any communicable disease. Unfortunately in the case of anthrax only too often the very first indication of the existence of the disease is the sudden death in the pasture or byre of an apparently healthy beast, or possibly of one or more sheep. Nevertheless, the importance of being able to recognise any early indications is very great, because an im- mediate and careful examination should at once be made of the stock on the farm, and suspicious cases isolated from the rest. The stock-man may notice that one or two animals tend to keep away from the others. They look dull and cease feeding, and possibly shivering may be observed. In horses swelling of the throat may occur, and in some places, there is discharge of blood from the orifices. Death follows the appearance of these symptoms in a few hours, and often with startling suddenness. Cattle die rapidly, but sheep, though rapidly contracting the disease, do not as a rule die so suddenly. 200 INFECTIVE DISEASES. The characteristic sign after death is enlargement of .the spleen to three or four times its natural size. It is not only -enlarged, but extremely soft and dark in colour. Blood spots are visible on the internal organs generally, and the intestine often contains a quantity of blood. The examination of a drop of blood will show under the microscope the characteristic bacilli. It is, however, quite unnecessary to make an elaborate post-mortem examination in order to satisfy oneself whether the disease is really anthrax or not. If an animal has died suddenly and has created a suspicion of anthrax, all that it is necessary to do is to cut cff an ear — or a foot in the case of a sheep — and make a cover-glass preparation at the first opportunity. A farmer with a case of anthrax must be made to realise the fact that an enormous quantity of poisonous material has to te dealt with. In fact, an infected animal is more dangerous when dead than alive. The owner or person in charge must immediately notify to a police constable the existence, or even a suspicion of the existence, of the disease. Prompt measures must be taken to destroy the carcass and all traces of the blood, and thus to reduce to a minimum the chance of the disease spreading to the rest of the stock, and of creating fresh outbreaks in the future. Every possible precaution must be taken to prevent the blood of the dead animal from contaminating the pasture, byre, or water supply. The rest of the stock should be removed from the pasture or cowshed where the disease has broken out. It is desirable to give a complete change of food and water, and the whole of the stock should be examined every day for a week, and any animals showing a rise of temperature should at once be isolated from the rest. Preventive inoculation has been recommended to protect the rest of the stock, but there is not sufficient evidence of the safety of the process to lead to the adoption of this treatment. Animals ready for the butcher may be removed from the risk of infection by immediate slaughter. To disinfect the pasture the best plan is a heavy top-dressing of lime, and after six weeks stock may be readmitted, though not without some risk. If year after year cases of anthrax occur on a particular pasture, the most obvious precaution is to keep stock from it altogether and convert it into arable land. As roots grown on anthrax- infected soil have been known to convey the disease, the wisest course if we have to deal with a small field or comparatively small tract of land is to throw it out of cultivation or to plant it with trees. ANTHRAX. 201 DISPOSAL OF THE CARCASS. The surest method to render harmless all the bacilli which exist in the carcass is burning, but cremation offers practical difficulties, especially if several carcasses have to be destroyed. In the case of an animal dying in a town, the local conditions may render it best to adopt destruction by burning or by means of chemicals. In such a case the carcass should be covered with quicklime, and then taken, in charge of an officer of the Local Authority, to a horse -slaughterer's or knacker's-yard, and destroyed by exposure to a high temperature, or by chemical agents especially in the vicinity of chemical works. Under the usual circumstances of death occurring on a farm, fortunately the simple plan of burial, with the addition of lime or other chemical agents, is perfectly efficacious, and even without the use of chemicals if the carcass has been left unopened, as the bacilli die rapidly if air is excluded. Some experiments carried out by M'Fadyean clearly indicate the importance of leaving the carcass unopened. On July 16th a sheep was infected with anthrax by feeding it with a virulent culture. Five days later it died, and a microscopic examina- tion of blood from the ear, immediately after death, showed very many anthrax bacilli. The carcass was left unskinned and unopened until July 27th, when the various organs were cut out of the chest and abdomen and placed in a tin box. The box was then buried at a depth of about two feet in garden earth, and left there undisturbed until February 15th, when it was exhumed. The organs had become con- verted into adipocere, and this was thoroughly mixed up with water and administered to a sheep. The sheep remained perfectly healthy. In another experiment a rabbit was inoculated with anthrax on June 1st. It died on June 3rd, and blood from the ear contained the bacilli. The rabbit was left unopened for three days, and then placed in a flower pot and buried in garden earth at a depth of two feet; It was exhumed on February 15th. The tissues were all destroyed by putrefaction, and the earth in contact with the bones was administered to a sheep without conveying the disease or producing any ill effects. Thus, in the first experiment, the lungs and the intestines, in which spore formation was most likely to occur, were used as a test, and in the second experiment the entire carcass. In both cases there was destruction or disappearance of the bacilli, and these tests, therefore, confirm in a very marked way the opinion that prompt burial of the unopened carcass is a perfectly safe plan to adopt. 202 INFECTIVE DISEASES. If an animal has died in a meadow, a pit six feet deep should be dug close to the carcass, and if quicklime can be procured with- out delay the carcass should be buried with a layer about a foot in depth beneath it and with about the same quantity to cover it, and the pit filled up with the excavated soil. If there are any traces of blood where the animal lay, the contaminated ground should be covered with quicklime or drenched with strong carbolic acid, and the whole of the site of burial fenced off for six months. If an animal dies near a brook or stream then the carcass must be removed for burial to a sufficient distance to prevent any reasonable probability of contamination of the water. If death has occurred in the byre, the carcass must be removed to the nearest and most convenient spot for burial, any fodder or litter which may have been in contact with the deceased animal must be destroyed, and the shed and cart and any utensils, hurdles, etc., disinfected. For the latter purpose thorough scouring with water and then washing with limewash is recommended. The limewash should be prepared immediately before use, and four ounces of chloride of lime, or half a pint of commercial carbolic acid, be added to each gallon of limewash. The following is an illustration of the value of preventive measures based upon a knowledge of the exact nature of the disease. A farm on the banks of the Yeo was repeatedly attacked by anthrax. One morning two sheep died, and other cases followed. The farmer learnt that his predecessor had buried cattle which had died of anthrax on the very spot where the sheep were folded. He removed his flock, and had no further losses among the sheep, but he continued to lose cattle grazing in the pastures by the river. These pastures were occasionally flooded by the Yeo. Another farmer in the same locality heavily manured a field, and shortly afterwards anthrax broke out in a most deadly form on his farm. What was the cause of these mysterious outbreaks ? The explanation was forthcoming, and prevention an easy matter. The river Yeo received the washings from the wool factories at Yeovil, and the pastures were contaminated by anthrax spores in the deposit which was left behind when the flood subsided. In the second instance, it was found that the manure used for dressing the pasture consisted of a quantity of refuse from the wool factories. Infected wool from foreign countries is one of the principal sources of the disease in this country, and the remedy is to insist upon the factories destroying their refuse instead of its being allowed to contaminate, the rivers or to be sold as manure. ANTHRAX. 203 So long as this source of the disease was unknown anthrax continued to be spread through the agency of the wool factories. Anthrax spores may also be introduced with foreign oats, hay, and manure, so that it is almost impossible absolutely to prevent the importation of the disease; but the danger of its unlimited extension and disastrous losses can be minimised, and the com- munication of the disease to man and to swine entirely avoided by simple precautions. ANTHRAX IN SWINE. The occurrence of anthrax in swine is a subject upon which there has long been considerable diversity of opinion. Some of the FIG. 99. — ANTHRAX IN SWINE. From a photograph taken during life, showing a swollen condition of the neck and throat six days after ingestion of part of the viscera of a bullock which had died from anthrax. earliest writers on the diseases of animals speak of outbreaks of anthrax among swine, but whether any or all of these outbreaks were examples of true anthrax has long been a matter of un- certainty; for it is well known that diseases quite distinct were included under the name anthrax. Menschel states that in an outbreak in which twenty-four persons were attacked with malignant pustule, many of them from eating the flesh of beasts suffering from anthrax, pigs which were fed on the same flesh also became affected, and a woman who ate some of the diseased pork was subsequently ill. Roche-Lubin, while apparently accepting the occurrence of anthrax in swine, taught that the pig resisted inoculation with the blood of ^ different species. 204 INFECTIVE DISEASES. In this country accounts have been published from time to time of a fatal disease in pigs induced by eating the flesh of animals which had died of what was described as " blood-poisoning." Some very striking cases occurred in the practice of Mr. Wilson, of Berkhampstead, and were reported in the Veterinarian. A farmer consulted Mr. Wilson respecting an illness with which his pigs were affected, stating that two or three were dead and many others seriously ill. They were strong hogs, ranging from six to nine months old. On inquiry it was ascertained that the farmer had lost a beast suddenly about a week previously, that the carcass had been opened in the yard, and the viscera thrown to the pigs. Mr. Wilson expressed the belief that the disease was anthrax, and stated that he found the pigs exhibiting many of the symptoms observable in cattle, with the additional one of enlargement round the throat from infiltration of a yellow fluid causing discoloration of the skin. Also, in the reports of the Agricultural Department of the Privy Council thirteen pigs were reported as suffering from anthrax in 1886, and one hundred and fifty-nine in 1887. But the question arose whether the disease in the pigs was genuine anthrax or septic poisoning. Williams says : " The flesh of animals which have died or have been killed whilst suffering from the disease [anthrax] should not be used as food either for men, pigs, or dogs, as it is apt to cause death by blood poisoning " ; and Steel writes : " Pigs, dogs, and poultry should not be allowed to feed on blood, flesh, and eject a of anthrax victims," but no statement is made as to the nature of the illness produced. No doubt these writers have been greatly influenced by the opinion of many bacteriologists, for Toussaint maintained that pigs could not be infected with anthrax, and a similar view was at one time upheld in this country by Klein, who stated that pigs were very insusceptible. In Germany also, pigs have been credited with an immunity from this disease. In the face of these conflicting statements the author carried out a series of experiments in order to ascertain the nature of the disease in swine resulting from the ingestion of the offal of animals which had died of anthrax ; and the result of inoculation with blood of animals which had died of anthrax, and with pure cultivations of the Bacillus anthracis. As a result of these experiments genuine anthrax was produced in swine (a) by feeding them with anthrax offal ; (b) by injection of blood of a bullock which had died of anthrax ; (c) by passing ANTHRAX. 205 bacilli through the guinea-pig, and transmitting them to swine by injection of blood from the spleen ; (d) by injecting a pure cultiva- tion of the anthrax bacillus; (e) and lastly, the anthrax bacillus was isolated from swine in which the disease was accidentally induced on a farm, and the disease reproduced by inoculation of guinea-pigs and mice with blood from the spleen. The Author's Conclusions. — Swine of all ages can be affected with anthrax. If the disease is induced by ingestion of anthrax offal, the tonsils are ulcerated, and constitute the point of access of the bacilli to the blood. In such cases the characteristic symptom is FIG. 100. — ANTHRAX IN SWINE. From a photograph taken post-mortem. Death occurred four days after the ingestion of offal from a bullock which had died of anthrax, and there was well-marked oedema of the throat, cheeks, and eyelids. enormous swelling around the throat. If the disease is induced by hypodermic injection, the same cedematous infiltration of the tissues occurs at the place selected for inoculation. Death may occur in twenty-four hours, or not until after five or six days. There is a rapid rise of temperature, usually a rash -like discoloration of the skin, sometimes loss of power over the limbs, and general weakness and disinclination to move ; the a nimal may lie helplessly on its belly, and utter plaintive cries when disturbed. At the post- mortem the most characteristic feature is the gelatinous oedema which, in the case of ingestion of offal, is found around the throat. There is usually congestion of all the organs and engorgement of 206 INFECTIVE DISEASES. the heart and large vessels, fluid in the cavities of the chest and abdomen, and enlargement and haemorrhage into the lymphatic glands. There is in some cases inflammation of the intestines with .submucous and subserous haemorrhages. The spleen may be normal in size, pale and flabby, and the liver only slightly congested and friable ; in other cases the condition is characteristic, the spleen is the seat of haemorrhage, causing more or less local enlargement, which is superficially of a deep purple colour ; the liver may also be greatly congested, very friable, and marked with purple patches. The examination of the blood of the heart and spleen for anthrax bacilli must be carried out with great perseverance and discrimi- nation, as they are present only in small numbers, and in some cases have given place entirely to septic organisms. Inoculation with the blood will produce either typical anthrax, or malignant cedema or some other form of septicaemia. Possibly in, the cases arising from ingestion of offal the ulcerated condition of the throat affords a nidus and a means of access for septic organisms. It is also well known that blood in a state of putrefaction may contain the bacillus of malignant cedema. In the presence of putrefactive organisms the anthrax bacillus rapidly disappears. If, therefore, inoculation of guinea-pigs or mice is used as a test for ascertaining the nature of an outbreak in swine, it must not be concluded, if Pasteur's or some other form of septicaemia result, that the disease was not anthrax, while, on the other hand, the discovery of the anthrax bacillus in the blood of the pig, or the production of anthrax in guinea-pigs or mice, is positive evidence as to the nature of the original disease. Peuch, in France, had obtained similar results by injecting pigs with anthrax blood and anthrax cultures. He also carried out some interesting experiments bearing on public health. The leg of a pig which had died of anthrax was covered with pounded sea-salt. Previously to the curing, a slice of the flesh was squeezed in a meat- press, and the liquid thus obtained was employed for inoculation. The animals inoculated died of typical anthrax. In six weeks the curing was considered to be completed, and a slice was cut from the ham and soaked in filtered water. The juice was extracted in the meat-press, and employed for the inoculation of four guinea-pigs and three rabbits. Slight swelling and a certain amount of redness at the seat of inoculation were the only results. A few drops of the muscle-juice were added to sterilised broth, and produced a mixed cultivation of micrococci and motile bacilli. A rabbit and two guinea-pigs inoculated with the cultivation remained quite healthy. ANTHRAX. 207 These experiments demonstrated that salting destroys the viru- lence of the flesh of pigs which have died of anthrax, but in order to obtain this result the salting must be thoroughly carried out. If the process be incomplete the flesh is still virulent. Thus the leg of a pig salted for only fourteen days furnished a juice which possessed a certain amount of virulence. Out of three inoculated rabbits, one died in ninety-seven hours of anthrax, and the others recovered. Three guinea-pigs all succumbed, and a fourth guinea- pig inoculated with a cultivation from the muscle-juice also died. Peuch considers that there is danger in consuming flesh which has not been thoroughly cured. As it has been clearly shown that pigs may become infected with anthrax, these animals come under the Anthrax Order of 1886. This provides for the disposal of the carcass; and although Peuch has shown that salting destroys the virulence of the flesh of pigs which have died of anthrax, there can be no doubt that it is quite right that such animals should be condemned as unfit for food. Further, the recognition of the occurrence of true anthrax in swine is an additional reason for condemning the Continental practice of eating hams, sausages, etc., in the raw state. Indeed, the viru- lence of anthrax flesh suggests one possible explanation of some of those obscure cases of meat poisoning which have occurred in this country. It is possible that the flesh of animals which had died of anthrax was used in the preparation of sausages, pork- pies, etc., and that the cooking was not sufficient to deprive the meat of its poisonous properties. EQUINE ANTHRAX. Veterinary authorities have described " Anthrax in the Horse," but it remains to be seen whether there are not two or more affec- tions included under this heading. Fleming says : " The most acute form of anthrax, the apoplectic, is somewhat rare in the horse, and has perhaps been most frequently observed on the Continent. Though cases are recorded, but through an error in diagnosis, under other names in the veterinary literature of this country, I have only witnessed two cases in England ; though during the intense summer heat in the north of China I had several." The question to which the author is in a position to give a definite answer is, whether the disease produced by the Bacillus anthracis ever occurs in the horse. Whether that has been pre- viously determined, at any rate in this country, it is difficult to say. 208 INFECTIVE DISEASES. Fleming in describing the pathological anatomy of anthrax in the horse, says : " The spleen is double and treble its ordinary volume ; its surface is sometimes bosselated by tumours; its texture is softened and transformed into a viscid reddish-brown or violet mass, and the mesenteric glands are infiltrated. The blood in it has been found to contain bacteridia when examined soon after death." Williams, who says that " anthrax in the horse rarely occurs in this country," adds, that it is prevalent in India, and is there termed " Loodiana disease," and in Africa " Horse-sick- ness." But " Horse-sickness," from recent researches, is certainly not anthrax. Williams described a case which occurred in 1879 as one of anthrax. A carriage-horse died suddenly while in harness ; " a large black tumour was found in the lungs, and the pulmonary arteries were engorged with black tarry blood, which, when micro- scopically examined, was found to contain the bacilli in a most perfect form, and very numerous indeed." In 1884, an outbreak of charbonous fever occurred in Liverpool. Williams proceeded to investigate the outbreak, and found two horses dead on his arrival, one having died only a few hours previously. The bacilli from the blood in this case are figured, and the following statement made : " These bacilli seem to differ from those of splenic fever, being rather smaller in diameter, and so far as my observations go, multiply by fission only, not developing spores." On the other hand, the author investigated the blood of a mare which was supposed to have died of anthrax, and on examining cover-glass preparations of the blood, it was found to contain large numbers of bacilli with the characteristic microscopical appearances of anthrax bacilli. To place the question beyond any possible doubt a number of tubes of agar-agar were inoculated. These, after three days in the incubator, produced typical cultivations, and on examina- tion by the ordinary methods and by double-staining, yielded very beautiful preparations of filaments and spores. At the same time that the cultivations were prepared, two mice were inoculated at the root of the tail with a trace of the blood. Two days afterwards they were both found dead, and with the characteristic post-mortem appearances, spleen much enlarged, and anthrax bacilli in enormous numbers. There can be no doubt that true anthrax occurs in the horse ; and the author, in 1887, recommended that it should be scheduled under the Contagious Diseases (Animals) Act, and equine anthrax has been included in the Anthrax Order of 1895. More recently Pemberthy has described cases of equine anthrax ANTHRAX. 209 which he believes to have been the result of infection from feeding on foreign oats or imported hay. Preventive Inoculation. — The prevention of anthrax by means of protective inoculation or vaccination has been attempted on a very large scale in France, and it is claimed that the results have been very beneficial to agriculture in that country : — Animals Mortality. Total Na. of Vacci- Total Average No. of Veteri- nated loss Ios7 Year. Animals Vacci- nated. nary Re- ports. after Receipt of Reports. After 1st Vacci- nation. After 2nd Vacci- nation. During rest of Year. Total. per cent. before Vacci- nation. /1S82 270..040 112 243,199 756 847 1,037 2,640 1-08 10% 1883 268,505 103 193,119 436 272 784 1,492 0-77 1884 316,553 109 231,693 770 444 1,083 2,247 0-97 1885 342,040 144 280,107 884 735 990 2,609 0-93 1886 313,288 88 202,064 652 303 514 1,469 0-72 1887 293,572 107 187,811 718 737 968 2,423 1-29 Sheep ^ 1888 269,574 50 101,834 149 181 300 630 0-62 1889 239,974 43 88,483 238 285 501 1,024 1-16 1890 223,611 69 69,865 331 261 244 836 1-20 1891 218,629 65 53,640 181 102 77 360 0-67 1892 259,696 70 63,125 319 183 126 628 0-99 ,1893 281,333 30 73,939 234 56 224 514 0-69 Total . 3,296,815 990 1,788,677 5,668 4,406 6,798 16,872 0-94 (0-32%) (0-24%) (0-38%) /1882 35,654 127 22,916 22 12 48 82 0-35 5% 1883 26,453 130 20,501 17 1 46 64 0-31 1884 33,900 139 22,616 20 13 52 85 0-37 1885 34,000 192 21,073 32 8 67 107 0-50 1886 39,154 135 22,113 18 7 39 64 0-29 Oxen 1887 48,484 148 28,083 23 18 68 109 0-39 or 1888 34,464 61 10,920 8 4 35 47 0-43 Cows 1889 32,251 68 11,610 14 7 31 52 0-45 1890 33,965 71 11,057 5 4 14 23 0-21 1891 40,736 68 10,476 6 4 4 14 0-13 1892 41,609 71 9,757 8 3 15 26 0-26 ,1893 38,154 45 9,840 4 1 13 18 0-18 438,824 1,255 200,962 177 82 432 691 0-34 (0-09%) (0-04%) (0-21%) i 14 210 INFECTIVE DISEASES. The vaccine is supplied, by a company in Paris, in two strengths. Reports are supplied by veterinary surgeons, and the results have been tabulated by Chamberland and published, and commented upon by Cope in a report to the Board of Agriculture (1894). The column of deaths, in the above table, includes the animals which died from the vaccination, and those which died from natural infection. It is claimed that the percentage of losses has been reduced from 10 per cent, to "94 per cent, in sheep, and from 5 per cent, to '34 per cent, in cattle. Cope, in the report just referred to, regards these conclusions as somewhat fallacious, because in order to prove that the animals inoculated received immunity, it should be shown that they were subsequently exposed to the risks of natural infection. This was not the case. But a report obtained from the Bureau in Paris gives the actual number of animals on each of the infected farms, and the number which have died of the disease ; and when compared with Chamberland's statistics it is evident that nine-tenths were not on farms where the disease appeared — at least, during 1889-92 — and that the deaths from anthrax on those farms where it was reported to exist were, if anything, higher than they were supposed to be prior to the introduction of the system of vaccination ; and in spite of the immense number of animals vaccinated the official returns obtained from Paris, by Cope, indicate that the mortality from anthrax, calculated in the ordinary way, remains as high as ever. Anthrax in France. Year. No. of Outbreaks re- ported. No. of Animals in Premises. No. of which Died. Percentage of Loss. 1889 618 22,599 1,458 6-5 1890 536 ' 24,073 1,123 4-7 1891 570 21,356 1,444 6-8 1892 607 28,199 1,581 5-6 CATTLE. 1889 — 6,059 700 11-6 1890 — 5,365 771 14-4 1891 — 7,299 849 11-6 1892 — 5,058 804 15-9 SHEEP. 1889 16,540 755 4-6 1890 18,708' 352 1-Sl 1891 14,057 545 4-2 1892 — 23,141 777 3-4 ANTHRAX. 211 In Germany, veterinary and agricultural authorities agree that the results have not met with the success which has been claimed for vaccination in France. Experiments were undertaken for the German Government, and in one set of experiments twenty-five sheep were vaccinated with the first vaccine without an accident, but three died five days after the second vaccine. In another experiment two hundred and fifty-one sheep were vaccinated with only one death, and subsequent inoculation with virulent anthrax proved that they had immunity. Six head of cattle were vaccinated without any loss, and six more were used for a control experiment. Inoculation with virulent virus proved fatal to the control animals, but the vaccinated were pro- tected. These, with other animals similarly vaccinated, amounting in all to two hundred and sixty-six sheep and eighty-three head of cattle, were then turned out to graze on infected pastures with two hundred and sixteen unvaccinated sheep as a control experiment. Within five months four of the vaccinated and eight of the un- vaccinated sheep died of anthrax, and one of the vaccinated and one of the unvaccinated cattle. The result of these experiments led to the following conclusions : — (1) That the first vaccine is mild and harmless. (2) That the second vaccine, even in the hands of experts, is dangerous and often fatal. (3) That sheep are more affected than cattle by the injections,, exhibiting fever and other indications of illness. (4) That cattle and sheep which recover from the vaccination have an immunity against anthrax when tested by experimental inoculation. (5) That vaccinated cattle and sheep tested by exposure to natural infection by grazing on infected pastures contract the disease in the ordinary way. (6) That the time for which immunity is conferred has not been determined. In England, Klein tested the vaccine, with the residt that animals either succumbed to the vaccine, or to virulent anthrax after recovery from the vaccine. Protective inoculation has also been employed in a few instances by leading agriculturists, but with very unsatis- factory results. Stamping -out System. — In Germany the conclusion is that the safest measures are destruction of carcasses and disinfection, and that inoculation will have no effect in lessening the loss caused by this disease. 212 INFECTIVE DISEASES. Iii England the stamping- out system has been advocated for many years, and is still regarded as the only reliable means for suppressing the disease ; and the possible introduction of the disease among healthy stock by vaccination, and especially in localities in which anthrax is unknown, would be contrary to the principles upon which the system is based. These principles are illustrated by the following extracts from the Anthrax Order of 1895 : — NOTIFICATION. 2. — (1) Every person having or having had in his possession or under his charge, an animal affected with or suspected of anthrax, shall, with all practicable speed, give notice of the fact of the animal being so affected or suspected, to a constable of the police force for the police area wherein the animal so affected or suspected is or was. (2) The constable shall forthwith give information of the receipt by him of the notice to an Inspector of the Local Authority, who shall forth- with report the same to the Local Authority. (3) The Inspector of the Local Authority shall forthwith give information of the receipt by him of the notice to the Medical Officer of Health of the Sanitary District in which the affected or suspected animal is or was. Duty of Inspector to act immediately. 3. An Inspector of a Local Authority on receiving in any manner whatsoever information of the supposed existence of anthrax, or having reasonable ground to suspect the existence of anthrax, shall proceed with all practicable speed to the place where such disease, according to the information received by him, exists, or is suspected to exist, and shall there and elsewhere put in force and discharge the powers and duties conferred and imposed on him as Inspector, by or under the Act of 1894 and this Order. Public Wanting as to Existence of Disease. 4. — (1) The Local Authority may, if they think fit, give public warning by placards, advertisement, or otherwise, of the existence of anthrax in any shed, stable, building, field, or other place, with or without any particular description thereof, as they think fit, and may continue to do so during the existence of the disease, and, in case of a shed, stable, building, or other like place, until the same has been cleansed and dis- infected in accordance with this Order. (2) It shall not be lawful for any person (without authority or excuse) to remove or deface any such placard. Milk of Diseased or Suspected Cow not to be Removed. 5. Where anthrax exists or has existed in any shed, stable, building, or other place, it shall not be lawful to remove from such shed, stable, ANTHRAX. 213 building, or other place the milk of any cow which is affected with or suspected of anthrax. Removal of Dung or other Things. 6. It shall not bei lawful for any person to send or carry, or cause to be sent or carried, on a railway, canal, river, or inland navigation, or in a coasting vessel, or on a highway or thoroughfare, any dung, fodder, or litter that has been in any place in contact with or used about a diseased or suspected animal, except with a Licence of the Local Authority for the District in which such place is situate, on a certificate of an Inspector of the Local Authority certifying that the thing moved has been, so far as practicable, disinfected. Disposal of Carcasses. 1. — (1) The carcass of an animal which at the time of its death was affected with or suspected of anthrax shall be disposed of by the Local Authority as follows : — (i.) Either the Local Authority shall cause the, carcass to be buried as soon as possible in its skin in some convenient or suitable place removed from any dwelling house and at such a distance from any well or watercourse as will preclude any risk of the contamination of the water therein, and at a depth of not less than six feet below the surface of the earth, having a layer of lime not less than one foot deep beneath, and a similar layer of lime above, the carcass ; (ii.) Or the Local Authority may, if authorised by Licence of the Board, cause the carcass to be destroyed, under the inspection of the Local Authority, in the mode following: The carcass shall be disinfected, and shall then be taken, in charge of an officer of the Local Authority, to a horse-slaughterer's or knacker's-yard approved for the purpose by the Board, or other place so approved, and shall be there destroyed by exposure to a high temperature, or by chemical agents. (2) With the view to the execution of the foregoing provisions of this Article the Local Authority may make such Regulations as they think fit for prohibiting or regulating the removal of carcasses, or for securing the burial or destruction of the same. (3) Before a carcass is removed for burial or destruction under this Article it shall be covered with quicklime. In no case shall the skin of the carcass be cut, nor shall anything be done to cause the effusion of blood. (4) A Local Authority may cause or allow a carcass to be taken into the District of another Local Authority to be buried or destroyed, with the previous consent of that Local Authority, but not otherwise. Digging Up. 8. It shall not be lawful for any person, except with the Licence of the Board or permission in writing of an Inspector of the Board, to dig 214 INFECTIVE DISEASES. up. or cause to be dug up, the carcass of any animal that has been buried. Disinfection in Case of Anthrax. 9. — (1) The Local Authority shall at their own expense cause to be cleansed and disinfected in the mode provided by this Article — (a) All those parts of any shed, stable, building, or other place in which a diseased or suspected, animal has been kept or has died or been slaughtered ; (b) Every utensil, pen, hurdle, or other thing used for or about any diseased or suspected animal ; (c) Every van, cart, or other vehicle used for carrying any diseased or suspected animal on land otherwise than on a railway. (2) The mode of the cleansing and disinfection of such shed, stable, building, or other place, or the part thereof, shall be as follows : — (i.) All those parts aforesaid of the shed, stable, building, or other place shall be swept out, and all litter, dung, or other thing that has been in contact with, or used about, any diseased or suspected animal shall be effectually removed therefrom ; then (ii.) The floor arid all other parts of the shed, stable, building, or other place with which the diseased or suspected animal or its droppings or any discharge from the mouth or nostrils of the animal has come in contact, shall be, so far as practicable, thoroughly washed or scrubbed or scoured with water ; then (iii.) The same parts of the shed, stable, building, or other place shall be washed over with limewash made of freshly burnt lime and water, and containing in each gallon of limewash four ounces of chloride of lime or half a pint of commercial carbolic acid, the limewash being prepared immediately before use ; (iv.) Except that where any place as aforesaid is not capable of being so cleansed and disinfected, it shall be sufficient if such place be cleansed and disinfected so far as practicable. (3) The mode of the cleansing and disinfection of such utensil, pen, hurdle, or other thing, and such van, cart, or other vehicle aforesaid, shall be as follows : — (i.) Each utensil, pen, hurdle, or other thing, van, cart, or other vehicle, shall be thoroughly scraped, and all litter, dung, sawdust, or other thing shall be effectually removed therefrom ; then (ii.) It shall be thoroughly washed or scrubbed or scoured with water ; then (iii.) It shall be washed over with limewash made of freshly burnt lime and water, and containing in each gallon of limewash four ounces of chloride of lime or half a pint of commercial carbolic acid, the limewash being prepared immediately before use. (4) All litter, dung, or other thing that has been removed from any such shed, stable, building, place, van, cart, or vehicle as aforesaid, shall be forthwith burnt or otherwise destroyed or disinfected to the satisfac- tion of an Inspector of the Local Authority. ANTHRAX. 215 (5) The Local Authority may make such Regulations as they think fit for the purpose of carrying out the provisions of this Article. Occupiers to give Facilities for Cleansing. 10. — (1) Where the power of causing any place, thing, or vehicle to be cleansed and disinfected under this Order is exercised by a Local Authority, the owner and occupier and person in charge of the place, thing, or vehicle shall give all reasonable facilities for that purpose. (2) Any person failing to comply with the provisions of this Article shall be deemed guilty of an offence against the Act of 1894. Regulations of Local Authority as to Movement of Animals, Fodder, etc. 11. A Local Authority may make such Regulations as they think fit for the following purposes, or any of them : — (a) For prohibiting or regulating the movement of any diseased or suspected animal into or out of any shed, stable, building, field, or other place, or any part thereof ; (6) For prohibiting or regulating the movement of any animal into or out of any shed, stable, building, field, or other place, or any part thereof, in which there is or has been any diseased or suspected animal ; and (c) For regulating the removal out of any shed, stable, building, field, or other place of any fodder, litter, or other thing that has been in contact with or used for or about any diseased or suspected animal ; but nothing in any such Regulation shall authorise movement in contravention of any provision of any Order of the Board for the time being in force ; and a Regulation under paragraph (6) of this Article shall operate so long only as any animal which in the judgment of the Local Authority is diseased or suspected remains in the shed, stable, building, field, or other place to which the Regulation refers, and, in case of a shed, stable, building, or other like place, until the same has been cleansed and disinfected in accordance with this Order. Slaughter hi Anthrax and Compensation. 12. — (1) A Local Authority may if they think fit cause to be slaughtered — (a) Any animal affected with anthrax or suspected of being so affected ; and (6) Any animal being or having been in the same field, shed, or other place, or in the same herd or flock or otherwise in contact with animals affected with anthrax, or being or having been in the opinion of the Local Authority in any way exposed to the infection of anthrax. (2) The slaughter of animals under this Article shall be conducted in such mode as will so far as possible prevent effusion of blood. 216 INFECTIVE DISEASES. (3) The Local Authority shall out of the local rate pay compensation as follows for animals slaughtered under this Article : — (a) Where the animal slaughtered was affected with anthrax the compensation shall be one-half of the value of the animal immediately before it became so affected ; and (5) In every other case the compensation shall be the value of the animal immediately before it was slaughtered. (4) Provided, that if the owner of the animal gives notice in writing to the Local Authority, or their Inspector or other officer, that he objects to the animal being slaughtered, it shall not be lawful for the Local Authority to cause that animal to be slaughtered except with the further special authority of the Board first obtained. Keeping of Swine in Slaughter-houses . 16. It shall not be lawful for any person, in any case in which the slaughter of any animal is authorised or required by this Order, to use for such slaughter any slaughter-house in which swine are kept. Whether an anthrax virus can be obtained which is absolutely incapable of creating centres of infection, and can therefore be recommended with safety for vaccination as an auxiliary and voluntary measure, is a matter for further investigation. CHAPTER XV. QUARTER-EVIL. — MALIGNANT (EDEMA. — RAG-PICKERS' SEPTICAEMIA. — SEPTICLEMIA OF GUINEA-PIGS.— SEPTICAEMIA OF MICE. QUARTER-EVIL in cattle, malignant oedema, and rag-pickers' septicaemia in man, septicaemia in guinea-pigs, and septicaemia in mice, are all varieties of septicaemia produced by bacilli. An account of quarter-evil, malignant oedema, and rag-pickers' septicaemia may appropriately follow the chapter on anthrax, as they have certain similarities to that disease. They are, however, not only distinct from anthrax, but must be carefully distinguished from each other. In connection with these forms of bacillary septicaemia in man and cattle we may study bacillary septicaemia in small animals. QUARTER- EVIL. The disease known in this country as quarter-evil or black-leg is identical with the French Charbon symptomatique and the German Rauschbrand. Symptomatic anthrax in a very slight degree resembles anthrax. The disease occurs usually in young cattle from a few weeks to about twelve months old, and attacks sheep and horses, but not swine or poultry. It is characterised by the develop- ment of an emphysematous swelling of the subcutaneous tissue and muscles, generally over the hind quarter. Infected animals cease feeding, the temperature rises, lameness supervenes, and death occurs in about forty-eight hours. The tumour on incision is found to contain a quantity of dark sanguineous fluid, with characteristic bacilli. Bacillus of Quarter-evil (Bacille du charbwi symptomatique, Rauschbrand bacillus).- — Motile rods with rounded ends, 3 to 5 p. in length, -5 to *6 p. in. breadth. Spore-formation present. The spores are oval, generally situated near the extremity of the rods, and when fully developed considerably exceed the rods in diameter. 217 218 INFECTIVE DISEASES. Involution forms are freely developed in old cultures, and in cultures made in unsuitable media. The bacilli possess numerous flagella, and their power of movement at <0fft once distinguishes them from anthrax bacilli. 0 4? They can be cultivated 1 « in the ordinary media o OM» P in the absence of oxy- ^ gen, but more readily with the addition of grape-sugar or glyce- rine. Radiating fila- f 9" ments grow out from ^ the more or less spher- ical colonies directly / o -* V •>'* FIG. 101. BACILLI OF QUAKTER-EVIL x 1000. From an agar culture (FRANKEL and PFEIFFER). liquefaction commences. In the depth of nutrient gelatine the growth occurs in two or three days at 20° to 25° 0. towards the lower part of the track of the inoculating needle. The gelatine slowly liquefies, and there is considerable formation of gas with the development of a peculiar odour. Spore- formation occurs freely in cultures, but not in the blood of infected animals until after death. Guinea-pigs inoculated with a pure- culture, or with spore-bearing threads, die in twenty-four to thirty-six hours. An em- physematous infiltration with sanguineous serum is produced at the seat of inoculation, and the surrounding muscles are of a dark colour. The internal organs are more or less Fl(, m PURE_CULTURE OF congested. The bacilli are found in the BACILLI OF QUARTER-EVIL local exudation and in the surrounding IN GRAPE-SUGAR GELA- TINE (FRANKEL PFEIFFER). tissue, and some hours surrounding after death in and QUARTER-EVIL. 219 increasing numbers in the blood of the heart and in the internal organs. Quarter-evil and malignant oedema, though possessing points of resemblance, are distinct diseases. Not only do the bacilli in the two cases differ in minute morphological and biological details, but Kitasato showed that guinea-pigs rendered immune against virulent quarter-evil had no immunity against malignant oedema. Protective Inoculation. — Arloing, Cornevin and Thomas have produced immunity by inoculating healthy cattle with a small quantity of the fluid from the tumour of an infected animal. Recovery takes place, and subsequent inoculation with a strong dose is without effect. Similar results may be obtained by intravenous injection of a few drops of the exudation. For general application of the system of protective inoculation, the virulent liquid and affected muscles are dried at 32° to 35° C., and the dried mass triturated with water and heated to 100°C. This is used as the first vaccine. An infusion similarly prepared, but only heated to 80° C., forms the second vaccine. The dry powder is a convenient form for general distribution, and -^ of a gramme is triturated with 5 cc. of water, and | cc. is injected into each animal. In about ten days the second vaccine is employed, and cattle so treated are said to have a complete immunity from fatal doses. The place chosen for the injection is the under surface of the tail, a short distance from the extremity. The hair is clipped at this spot, and the point of a syringe is pushed in between the skin and the bone, and the vaccine slowly injected. Eoux and Chamberland produced immunity by inoculation of filtered cultures. Cultures in broth were deprived of bacilli by heating to 115° C., or by filtration through porcelain. Guinea-pigs were inoculated with three doses of 30 cc. at intervals of two days, and subsequently injected with a solution of virulent black-leg powder and lactic acid, which killed control animals in twenty-four hours. Kitt employed cultures on agar a fortnight old, or fresh cultures sterilised by steam for thirty minutes. It was found possible to confer immunity in oxen, sheep, and guinea-pigs against the most virulent extract. Kitt's method has the advantage over others of only necessitating one single injection. Whether these experiments are of scientific interest rather than of practical value may be regarded as an open question. On the Continent, and especially in France, vaccination against quarter-evil has been carried out extensively ; and by comparing the 220 INFECTIVE DISEASES. mortality among the vaccinated and unvaccinated in localities where the disease commonly occurs, it has been said that the results are extremely favourable. The matter was investigated in this country by a committee of the Midland Veterinary Medical Association, and in the course of the experiments some surprising results were obtained. Six calves and four sheep were vaccinated, and five calves and two sheep were left unvaccinated as a control experiment. The seventeen animals were subsequently inoculated with virulent virus in the form of dried and powdered muscle. In forty- eight hours all the sheep died, and all the calves exhibited a swelling at the seat of inoculation. In another set of experiments, healthy calves inoculated with fresh juice from the tumour in a case of quarter-evil were not materially affected. The possibility of those calves which possess a natural immunity being classed as protected by the inoculation must be admitted, and the efficacy and safety of the process is by no means established. MALIGNANT CEDEMA. The disease known by surgeons as progressive gangrene, gan- grenous emphysema, or surgical gangrene, has been shown by the researches of Chauveau, Arloing, Rosenbach and Babes, to be due to a bacillus identical with the microbe septique of Pasteur and the bacillus of malignant oedema of Koch. The bacillus or its spores may be spread by the neglect of antiseptics. The disease occurs especially after compound fractures and gun-shot wounds. If a guinea-pig is subcutaneously inoculated with earth, putrid fluid, or hay dust, death frequently occurs in from twenty-four to forty- eight hours. At the autopsy the most characteristic symptom is a widespread subcutaneous oedema accompanied by air-bubbles. This originates from the point of inoculation, and contains a clear reddish liquid full of motile and n on- motile bacilli. The internal organs are little changed, the spleen is enlarged and of a dark colour, and the lungs are hypersemic, and have hsemorrhagic spots. Examined immediately after death, few or no bacilli are detected in the blood of the heart, but in that of the spleen, liver, lungs, and other organs, in the peritoneal exudation, and in and upon the serous coating of the abdominal organs, they are present in large numbers. If, on the other hand, the animal is not examined until some time after death, the bacilli are found in the blood of the heart, and distributed all over the body. Bacillus (Edematis Maligni, Koch (Pasteur's Septicaemia). — MALIGNANT (EDEMA. 221 Rods from 3 to 3'5 /*, long and 1 to 1*1 /x wide ; they mostly lie in pairs, and then appear to be double this length. The rods are rounded at their ends, and form threads which are sometimes straight, but more commonly curved. In stained preparations they have a somewhat granular appearance. They are motile, possessing flagella, and form spores. The bacilli are distinguished from anthrax bacilli by their being somewhat thinner, by their rounded ends, and by their motility. Moreover, anthrax bacilli never appear as threads in fresh blood, and are differently distributed throughout the body. They nre anaerobic, and can be cultivated on blood serum and on neutral solution of Liebig's meat extract in an atmosphere of carbonic acid. By embedding material containing bacilli in nutrient agar-agar \ FIG. 103. BACILLI OF MALIGNANT (EDEMA x 950. From the subcutaneous tissue of a guinea-pig. (BAUMGARTEN.) and nutrient gelatine, characteristic cultivations are obtained. The following process may be adopted to obtain a pure cultivation. A mouse inoculated subcutaneously with dust, as a rule, dies in one to two days. It is then pinned out, back uppermost, 011 a slab of wood, and the hair singed with a Paquelin's cautery from one hind leg up to the neck, across the latter, and down again to the opposite hind leg. Following the cauterised line, the skin is cut through with .sterilised scissors, and the flap turned back and pinned out of the way. With curved scissors little pieces of the subcutaneous cedematous tissue, in the neighbourhood of the inoculated spot, are cut out, and sunk with a platinum needle in a 1 per cent, nutrient agar-agar, or 5 per cent, nutrient gelatine. Fragments of tissue may also be embedded by the method already described for anaerobic bacteria. 222 INFECTIVE DISEASES. The inoculated tubes are placed in the incubator. In a few hours a whitish turbidity spreads out from the piece of tissue, and upwards in the needle track. Examined microscopically, the turbidity is found to be due solely to the development of the bacilli of oedema. The surface exposed to the air exhibits no trace of the bacilli. To investigate the tubes microscopically, a sterilised glass tube with a capillary end may be used, with its neck plugged with sterilised cotton -wool, and provided at the mouth with a suction ball. The capillary end is thrust into the cultiva- tion, and a small fragment removed by aspiration. In the course of the first day the bacilli spread throughout a great part of the agar-agar in such a way that a more or less equally diffused cloudiness of the medium en- sues, with subsequent appearance of strongly marked clouds or lines of turbidity. At the same time gas- bubbles develop along the needle track, and a collection of liquid takes place, while spore-formation also commences. The following day these appearances are more marked, the opacity is more pronounced, the development of gas increases, and the liquid contains more spore-forming bacilli and nu- merous free-spores. The nutrient -gelatine cultures during the first day show no macroscopic change, but FIG. 104. "PURE-CULTURE after a few days the piece of tissue is sur- OF BACILLUS OF MALIG- rounded with a white halo. This gradually spreads in all directions, and is apparently beset with hairs. The gelatine liquefies, and the fragment of tissue, degenerated bacilli,. and spores, sink to the bottom. The cultivation is also very characteristic in a ^ per cent, nutrient agar-agar. If placed in the incubator, in a few hours a cloudiness forms around the piece of embedded tissue, which is caused by bacilli gradually spreading in all directions in the nutrient medium. Mice inoculated from these cultivations die more quickly than from the original infection from dust. On potatoes they are cultivated by introducing a piece of liver or other tissue containing the bacilli, into the interior of a sterilised potato, and incubated at 38° C. The bacillus is not deprived of its virulence by cultivation. NANT (EDEMA IN GRAPE- SUGAR GELATINE (FRAN- KEL and PFEIFFER). j / MALIGNANT (EDEMA. 223 The spores of the oedema -bacilli appear to be very widely dis- tributed. They are found in the upper cultivated layers of the soil, in hay dust, in decomposing liquids, and especially in the bodies of suffocated animals, which are left to decompose at a high temperature. From any of these sources animals can be successfully inoculated. The bacillus is not only pathogenic in guinea-pigs, rabbits, and mice, but also in man and in farm animals, including calves but not cattle. Pure -cultures inoculated in animals produce oedema at the seat of inoculation without appreciable gas formation and without any putrefac- tive odour. The odour * and frothy effusion resulting from the in- oculation of earth are / ^ .^ due to other bacteria, \i which are introduced f simultaneously with X*~ / 4^ the bacilli of malig- \ ^ V iiant cedema. The _^ • ^^^ spleen is sometimes slightly enlarged. By touching with a cover- glass the capsule of FlG> m BACILLI OF MALIGNANT (EDEMA x 1000. the spleen, or by ex- From an agar culture (FBANKEL and PFEIFFER). amining the serous effusion, the bacilli are found in abundance ; but if a preparation is made from the interior of the spleen or from the blood of the heart, no bacilli will be found until several hours after death. In this respect there is a marked difference from anthrax. Another difference is shown in spore-formation, which occurs in the living body in malignant oedema, but never in anthrax. Animals which recover from the disease are said to be protected. Protective Inoculation. — Roux and Chamberland produced immunity by injecting the chemical products in the nitrate obtained from cultures in broth. The serum from fatal cases will, it is said, confer immunity on other animals. There is a variety of this bacillus in soil according to Fliigge, agreeing in morphological and cultural but not in pathogenic characters. 224 INFECTIVE DISEASES. RAG-PICKERS' SEPTICAEMIA. Hag-pickers' disease has a resemblance to anthrax or wool-sorters' disease. After death the spleen is found to be enlarged, the in- ternal organs are congested, and there are haemorrhages on the serous membranes. Bordoni -Uffreduzzi isolated bacilli which are quite easily distinguished from anthrax bacilli. They were found in the blood and in sections of the internal organs. Proteus hominis eapsulatus. — Rods with rounded ends, singly, in pairs, and in filaments, somewhat smaller than anthrax bacilli, and often irregular in form. Spore-formation not described ; they have a well-marked capsule. Colonies are circular, appearing at first granular, and later possessing a filamentous structure. In the depth of gelatine they grow in the shape of a round-headed nail, like a culture of Friedlander's pneumococcus. On the surface of gelatine they form a shining white layer. On agar the growth is somewhat transparent. On potato a moist, glistening film gradually spreads over the surface. They do not liquefy blood serum, and the growth is similar to that obtained on agar. They prove fatal to mice and dogs, but rabbits and guinea-pigs are not very sus- ceptible. Dogs die usually on the second day after intravenous injection, and after death there is congestion of the internal organs and of the intestinal mucous membrane. (Edema is produced at the seat of inoculation in mice. There are haemorrhages in the lymphatic glands, and congestion of the liver and kidneys. Similar organisms have been described by Kolb and by Babes in purpura hsemorrhagica. SEPTICAEMIA OF GUINEA-PIGS. Guinea-pigs and mice sometimes die of septicaemia, characterised by congestion of the lungs, liver, and kidneys, inflamed peritoneum, pleural and pericardial exudation, congested spleen, and congestion of the mucous and serous coats of the intestine. Klein isolated a bacillus from the blood and the internal organs in these cases. Bacillus of Septicaemia in Guinea-pigs. — Rods with rounded ends, motile, with pleomorphic forms, cocci, short rods and filaments. Colonies appear as small, circular, white dots, which enlarge and become irregular in outline. In the depth of gelatine a white filament develops, and on the surface the growth rapidly spreads with a crenated outline. Broth becomes turbid, and after the second day a copious white sediment is deposited. Spore-formation not observed. DESCRIPTION OF PLATE VI. Bacillus Murisepticus, FIG. 1. — From a section of a kidney of a mouse which had died after inocula- tion with a pure-cultivation of the bacillus. With moderate amplification, the white blood-corpuscles have a granular appearance, and irregular granular masses are scattered between the kidney tubules. Stained by Gram's method with eosin. x 200. FlG. 2. — Part of the same preparation with high amplification. The granular appearances are found to be due to the presence of great numbers of extremely minute bacilli, x 1500. Plate VI >- > i < . i. i Ce Figl. V, > I Fig, 2. BACILLUS MURISEPTICUS SEPTICAEMIA OF MICE. 225 According to Wooldridge, the chemical products of this bacillus, separated by filtration, produce on inoculation immunity against virulent bacilli. SEPTICAEMIA OF MICE. Mice inoculated with a minimum quantity of putrid fluid often die of septicaemia. They rapidly sicken, their eyes inflame, their eyelids stick together, they become soporific, and death occurs in forty to sixty hours. There is slight oedema at the seat of inoculation, and enlargement of the spleen ; the bacilli are found free and in the interior of white corpuscles, both in the cedematous tissue and in the blood capillaries. Bacillus of Septicaemia of Mice (Koch). —Extremely minute bacilli, -8 to 1 p long, and •1 to -2 fj. broad, and filaments. In cultivations in gelatine they do not appear to make threads, but the bacilli lie together in masses. Spores have been observed. The bacilli are probably non-motile. They are most commonly in the interior of white blood corpuscles. In these they increase, and in many cases a white blood cell is represented only by a mass of bacilli. A minimal quantity of blood containing the bacilli produces the disease if inoculated in house-mice or sparrows. Field-mice have an immunity. Rabbits and guinea-pigs inoculated in the ear suffer only from a local erythema, , J J ' FIG. 106.— PURE CUL- which disappears after five or six days, and TIVATION OF THE renders them for a time immune. Rabbits BACILLUS OF SEPTI- inoculated in the cornea suffer from an intense C^MIA OF MICE IN . a ,. » ,, mi i_ -IT .e • NUTRIENT GELATINE. inflammation of the eyes. The bacilli form m J After two days. plate-cultivations scarcely perceptible cloud-like specks, and in a test-tube of nutrient gelatine they form a delicately clouded cultivation along the needle track. An identical bacillus has been isolated in swine measles. 15 (CHAPTER XVT. SEPTICEMIA OF BUFFALOES. SEPTIC PLEURO-PNEUMONIA OF CALVES. — SWINE FEVER.— SEPTICAEMIA OF DEER.— SEPTICAEMIA OF RABBITS. — FOWL CHOLERA. — FOWL ENTERITIS. — DUCK CHOLERA. — GROUSE DISEASE. THERE are several varieties of septicaemia occurring naturally in buffaloes, deer, calves, and birds, and artificially induced by inocula- tion of rabbits with septic material. They are associated with bacteria which agree in their morphological and cultural characters, though in some cases differing in their pathogenic properties. As the differences between the bacteria cultivated from these different sources is not greater than the differences which exist between the morphological, biological, and pathogenic effects of varieties of the tubercle bacillus, it will be convenient and fully justifiable to follow Hueppe and Baumgarten, and regard them as varieties of the bacillus of hcemorrhagic septicaemia. EPIDEMIC DISEASE OF BUFFALOES. Oreste and Armanni investigated an epidemic among herds of young buffaloes in Italy (Eilffd-seuche). The disease was extremely acute, death occurring in from twelve to twenty -four hours. It was probably identical with an epidemic disease described by Bellinger in deer. The symptoms were fever, rapid pulse, discharge of mucus from the nose and mouth, and a local swelling of the head and face leading to suffocation. The only marked feature after death was hsemorrhagic inflammation of the small intestine. The bacilli were identical with those found by Schiitz in swine fever. Cultures inoculated in young buffaloes produced the disease. The bacilli were pathogenic to mice, guinea-pigs, rabbits, pigeons, and fowls, death taking place in from one to three days. 226 EPIDEMIC DISEASE OF DEER AND BOARS. 227 SEPTIC PLEURO-PNEUMONIA IN CALVES. Septic pleuro-pneumoriia is a disease which attacks young calves within the first two months after their birth. Percussion and auscultation reveal lung mischief. The disease is very rapid and fatal, death occurring on the second or third day. In the less acute cases one or more lobes of the lungs are found after death in a state of lobular and inter-lobular pneumonia. The inter-lobular connective tissue is distended with exudation, giving rise to white or yellowish bands between the inflamed lobules, which produce a marbled appearance, recalling the condition of the lungs in infectious pleuro-pneumonia. The internal organs are congested, and there are very often haemorrhagic spots on the mucous and serous coats of the small intestine. All the organs contain rods identical with those of septicaemia of rabbits. Rabbits, guinea-pigs, and mice were infected. A calf was injected in the pleural cavity with a broth - culture, and died in twenty hours. SWINE FEVER. This disease will be described in a separate chapter. Several bacteria have been isolated by different investigators. In swine fever in Germany (Schwein-seuche) Loffler and Schiitz isolated a bacillus which has been identified with the bacillus isolated by Salmon and Smith from hog-cholera in America, and with the bacillus of rabbit septicaemia and of fowl cholera. EPIDEMIC DISEASE OF DEER AND BOARS. A very fatal epizootic (Wildseuche) occurred in the royal game preserves near Munich, destroying one hundred and fifty-three deer and two hundred and thirty-four boars (Bellinger). The disease lasted from twelve hours to six days. In the less acute cases pneu- monia and pericarditis supervened. In cattle there was also severe haemorrhagic inflammation of the small intestine. In another form it produced swelling of the head, face, neck, and tongue. The virus proved fatal to rabbits in six to eight hours, and to sheep and goats in about thirty hours. A pig inoculated with a few drops of blood died in twenty-two hours. Kitt also investigated this malady. The bacteria were found to be identical, in their appearance and pathogenic properties, with extremely virulent bacteria from swine fever. Schiitz distinguished them from the bacteria obtained from swine fever by their pathogenic effect on pigeons, but cultures obtained from swine fever do not act uniformly in this respect. 228 INFECTIVE DISEASES. SEPTICAEMIA IN RABBITS. Koch minutely investigated a disease of rabbits produced by inoculation with impure river water and with putrid meat infusion. Bacteria are found in the blood in abundance, and may be readily cultivated. The smallest quantity inoculated subcutaneously or in the cornea of a rabbit produces a rise of tempera- ture and laboured breathing after ten to twelve hours, and death in sixteen ^^ ^ f ^ to twenty hours. The spleen and lymphatic glands are found to be FIG. 107. -BACTERIUM OF RABBIT enlarged, and the lungs congested, SEPTICAEMIA; BLOOD OF SPAR- ° . ' ROW, x 700 (Kocn). Dut there are no extravasations, and no peritonitis. Mice and birds are very susceptible ; guinea-pigs and white rats have an immunity. DAVAINE'S SEPTICAEMIA. A disease was produced by Davaine by injecting rabbits with putrid blood. Rabbits, mice, fowls, pigeons, and sparrows are sus- ceptible, and guinea-pigs and rats are insusceptible to the bacteria found in this disease. Rabbits inoculated with a trace of blood con- taining the bacteria, or with a culture, died in from twenty-four to thirty-six hours. The spleen, liver, lungs, and intestines are highly congested, and sometimes extravasations and peritonitis are found. FOWL CHOLERA. Fowl cholera is an epidemic disease of the poultry-yard much dreaded in France, and well known through the researches of Perroncito, Toussaint, Pasteur, and Kitt. •*• ,-, FIG. 109. — BACTERIUM OF FOWL 1 ™ ^ BAC™Mf°F F7L °*OL1f *» CHOLERA, x 2500. Muscle juice x 1200. From blood of inoculated Fowl. Fowls suffering from the disease usually die in from twenty-four to forty-eight hours. The disease shows itself by the fowls becoming FOWL CHOLERA. 229 somnolent. They suffer from weakness of the legs, and their wings trail. There is frequently diarrho3a, with slimy greenish evacuations, and death usually ensues after a slight convulsive attack. On making a post-mortem examination the viscera will be found to be congested, and there is intense inflammation of the mucous membrane of the intestine, with haemorrhages. The blood from the heart, and the intestinal contents, contain the bacilli which were at one time believed to be peculiar to this disease. Inoculation subcutaneously, or administration with food, of a small quantity of a broth cultivation will produce death in twenty-four to thirty-six hours. Pigeons, pheasants, sparrows, fn ^fcf^3;^i ^ffs?''^" FIG. 110. — BACTERIUM OF FOWL CHOLERA. Section from liver of Fowl x 700 (FLUGGE). rabbits, and mice are susceptible. In guinea-pigs, sheep, and horses, an abscess develops at the seat of inoculation. Rabbits are readily infected by sprinkling a broth-cultivation on cabbage leaves or any suitable food. It was with this microbe that Pasteur proposed to eradicate the plague of rabbits in Australia. Fowl cholera has an additional interest, as it was with this disease that Pasteur first investigated the attenuation of virus. Broth -cultures which were several months old were found, when injected, to produce apparently only a local effect. This weakening of the virus was attributed by Pasteur to exposure to oxygen. After recovery the fowls were protected against the action of virulent cultures, while fowls not immunised died the following day. Kitt, by working with pure-cultures on solid media, showed that the X^TBRATT?^ [ " OF THK *N I UNIVERSITY 1 230 INFECTIVE DISEASES. weakening was not due to prolonged exposure to oxygen, but that old contaminated broth- cultures after a time completely lost their power, owing to the antagonism of the bacteria accidentally present. Filtered broth- cultures contain the toxic products of the bacillus, and produce slight illness and subsequent immunity. FOWL ENTERITIS. Fowl enteritis is an acute infectious disease of fowls, the course and symptoms of which are regarded by Klein as distinct from fowl cholera. The fowls suffer from diarrhoea, with liquid greenish evacuations, but are never somnolent, and death occurs in one or two days. After death the mucous membrane of the intestine is found to be congested, and coated with grey or yellowish mucus; the liver is congested, spleen enlarged, and lungs normal. There are a few bacilli in the blood of the heart, very many in the spleen and liver, and they are in the form of a pure-culture in the mucus of the intestine. Klein says that the bacilli are a little longer and thicker than those found in fowl cholera, which they only slightly resemble, and that the course of the disease, the symptoms and pathological appearances, definitely distinguish it from fowl cholera, but that nevertheless it belongs to the same family of bacilli Pigeons are said to be insusceptible, rabbits only slightly susceptible. By feeding and by subcutaneous inoculation the disease can be communicated to healthy fowls, but there is no sign of illness until the fourth day. As regards attenuation, the bacilli behave like those from cases of fowl cholera. DUCK CHOLERA. Duck cholera is an epidemic disease of ducks which was investi- gated by Cornil. The symptoms are similar to those of fowl cholera. They suffer from diarrhoea and weakness, followed by death in two or three days. The bacillus cultivated from the blood of ducks is pathogenic in ducks but not in fowls or pigeons, and large doses are required to kill rabbits. GROUSE DISEASE. Grouse disease is an acute infectious disease of red grouse. According to Klein the chief pathological feature is severe pneu- monia ; there is also patchy redness of the serous and mucous linings GROUSE DISEASE. • -Y O juv FIG. 111.— BACILLUS OF H^MORRHAGIC SEPTICAEMIA. Blood of a Rabbit after death from Septicaemia x 950 (BAUM- GARTEN). of the intestine, and the liver is congested and dark, but the spleen is not enlarged. The bacilli are found in the heart, lungs, and liver, and in the extravasated blood. Cultures inoculated in mice and guinea-pigs produce pneumonia and death. Sparrows are sus- ceptible, and other small birds. Fowls, pigeons, and rabbits are insusceptible. Bacillus of Haemor- rhagic Septicaemia. — Very short rods, with rounded ends, *6 to '7 JJL in width and 1'4 //. in length. In stained preparations the rods are observed to be deeply stained at the ends and to have a clear interval in the middle; they were on this account mis- taken by earlier observers for dumb-bell micrococci or diplococci. They are non- motile, and spore-formation is unknown. They grow readily in the ordinary media. The colonies in nutrient gelatine appear about the third day. They are circular in form, with a sharp dark outline, and of a yellow colour, lighter at the peri- phery. Later, the central zone is finely granular, and of a dark yellowish-brown colour, with the lighter peripheral zone more clearly defined. In the depth of gelatine a delicate filament develops in the track of the needle, composed of minute spherical colonies, somewhat trans- parent, and yellowish-white in colour. At the point of puncture there may be no growth visible, or a flat and very limited growth. Inoculated on the surface of nutrient media a thin layer develops, with an irregular serrated and thickened border. On potato different results have been obtained by different observers. Some maintain that a greyish-white or yellowish film will develop at the temperature of the'blood ; but according to Caneva, the bacilli, whatever their source, FIG. 112. — BACILLUS OF H.EMORRHAGIC SEPTICAEMIA (Rabbit Septicaemia). Pure- culture in Gelatine after four days (B AUMG ARTEN. ) 232 INFECTIVE DISEASES. will not grow on potato, while Bunzl-Feclern maintains that the bacilli from fowl cholera and rahbit septicaemia do grow upon potato, but those from septicaemia in deer, buffaloes, and swine do not. Opinions differ with regard to their action on milk. The reaction for phenol and indol is given in all cases, except with cultures obtained from septicaemia of buffaloes. The virulence of the bacilli may be diminished and attenuated, but it may subsequently be restored by successive inoculation in animals. The pathological lesions vary in different animals. The most common result is con- gestion of the internal organs and haemorrhage. The bacilli culti- vated from cattle or deer produce fatal results when inoculated in swine. The bacilli from any of these sources inoculated in pigeons will produce fowl cholera, but the bacilli isolated by Schiitz from swine, and those from deer, are not fatal to fowls. Further, the bacilli cultivated from swine fever are fatal to guinea-pigs, while the bacilli from rabbit septicaemia have very little effect upon them. The bacilli have been found in association with diseases of cattle, swine, deer, birds, rabbits, and mice, and have been cultivated from healthy mucous membrane. Yeranus Moore found the bacilli in the mucus from the upper air passages, of 71 per cent, of cattle, 85 per cent, of cats, and 33 per cent, of dogs. From these sources inoculations were made in rabbits, and rapidly fatal septicaemia was produced, associated in less acute cases with peritonitis, pleurisy, and pericarditis. CHAPTER XVII. PNEUMONIA. — INFECTIOUS PLEURO-PNEUMONIA OF CATTLE. — INFLUENZA. ACUTE CROUPOUS PNEUMONIA. PNEUMONIA is an acute inflammation of the lungs with fibrinous infiltration of the air vesicles and interstitial tissue. There are varieties of pneumonia, and one form is commonly believed to be infectious. The lung passes through three stages — engorgement, red hepati- sation, and grey hepatisation. In the first stage the lung is of a deep red colour, but still vesicular ; in the second stage the affected part is more or less solid, and has the consistency of liver, owing to the fibrinous lymph which is poured out into the alveolar cavities. In the grey hepatisation, the exudation contains more leucocytes and less fibrin, and this is followed by the stage of suppurative softening and final absorption. The sputum at the commencement of the disease is rusty, from the presence of blood, and later on has the appearance of prune juice. Examination of the sputum by Gram's method will reveal numerous micro-organisms, and two of these are deserving of special study — the pneumococcus of Friedlander, which is present in a considerable proportion of cases, and Sternberg's micrococcus, which was found in sputum by Talamon. In 1888, there was considerable prevalence of pneumonia in Middlesbrough, with strong tendency to occur in groups of cases; but there was admittedly room for doubt whether the clinical and post-mortem appearances were not identical with ordinary pneumonia. Dr. Ballard maintained that there were facts and considerations which appeared to show that the disease was com- municable from the sick to the healthy, and that it was a specific febrile disease, and Klein isolated and described the micrococcus present in these cases. Bacterium Pneumonias Crouposae (Pneumococcus, Fried- 234 INFECTIVE DISEASES. FIG. 113.— BACTERIUM PNEUMONLE CROUPOS^E, FROM PLEURAL CAVITY OF A MOUSE, x 1500. A, B. Thread-forms. C, D, E. Short rod-forms. G. Diplococci. H. Cocci. I. Streptococci. (Zopf.) lander). — Cocci ellipsoidal and round, singly, or in pairs (diplococci), rods and thread-forms. The cell- membrane thickens, and develops into a gelatinous capsule, which is (round if the coccus is single, and ellipsoidal if the cocci occur in pairs or in rod-forms. Cultivated in a test-tube of nutrient gelatine they grow in the form of a round- headed nail, without liquefaction of the gelatine (Fig. 114). The cocci when artificially cultivated have no capsule, but it again appears after their injection into animals, The cocci can also be cultivated on blood serum and on boiled potatoes. They occur in pneumonic exudation. In- oculation of dogs with a cultivation of the cocci occasionally gave positive results; but in rabbits no results followed. Guinea- pigs proved to be susceptible in some cases ; but thirty- two mice, after injection of a cultivation diffused in sterilised water, into the lungs, died without exception. The lungs were red and solid, and contained the cocci, which were also present in the blood, and in enormous numbers in the pleura! exudation. Inhalation experiments by spray- ing the cocci diffused in water into mouse cages produced pneumonia and pleurisy in three out of ten mice. The nail-shaped cultivation is not always produced, nor are these conclusions accepted — - IIG. 114. — FBIEDLANDBBS by all investigators. METHODS OF STAINING FRIEDLANDER's PNEUMOCOCCUS. PNEUMOCOCCUS. Pure- culture in nutrient- gelatine four days old (BAUMGARTEN). Cover-glass preparations of pneumonic sputum or exudation may be treated as follows :— PNEUMONIA. 235 (a) (b) brown. balsam. (c) hours ; ( » FIG. 123.— BACILLI OF PLAGUE AND PHAGOCYTES, x 800. From human lymphatic gland. (AOYAMA.) congested and enlarged. In less acute cases an abscess of the abdominal wall occasionally results. The bacilli are sometimes found in the pleural and peritoneal exudation. The liver and spleen also contain many bacilli. Those in the blood are a little longer than those in the lymphatic glands. Inoculations can readily be made from guinea-pig to guinea- pig by using the pulp of the spleen, or the blood. Cultures lose their virulence gradually, but the virus can be intensified by successive inoculations in animals. The disease is infectious to mice as well as inoculable. Pigeons are insusceptible. Rats and flies may convey the bacilli. According to Aoyama, the bacilli found in the blood of plague 254 INFECTIVE DISEASES. patients and in the buboes are not identical. The bacilli in the buboes are different in form, and they stain by Gram's method. There is no doubt that the micro-organism which was found in blood is very similar to the bacillus of fowl cholera, and it is quite possible that the so-called plague bacillus is really identical with the bacillus of hsemorrhagic septicaemia, and that the real nature of the contagium in bubonic plague is unknown. Protective Inoculation. — Yersin, Calmette, and Borrell claim not only to have produced immunity, but to have cured animals after infection. Cultures on agar heated to 58° C. for an hour were attenuated, and rabbits after intravenous or subcutaneous inoculation were protected against virulent cultures. The serum of immunised rabbits was capable of protecting from subsequent virulent cultures, and neutralising the effect of a previous inoculation of a virulent culture. A horse was inoculated with cultures which killed mice in two days, and after six weeks a serum was obtained which produced immunity in mice and guinea-pigs. Stamping-out System.— It is not until the sixteenth century that we hear of preventive measures being attempted in England, and then they appear to have been adopted only when an outbreak threatened to be very serious. Early in the sixteenth century all those who had the plague in their houses were ordered to put up wisps, and to carry white rods in their hands. In 1543 the Plague Order of Henry VIII. was issued. In place of wisps the sign of the cross was to be made on every infected house, and to remain there for forty days. Persons afflicted with the disease were to refrain, if possible, from going out of doors, or for forty days to carry a white rod in the hand. All straw from the infected houses was to be carried into the fields and burnt. Churchwardens were directed to keep beggars out of churches on holy days, and all streets and lanes were to be cleansed. In 1547 the means of notification was a blue cross with the addition of the inscription Lord have mercy upon us. Later on the colour of the cross was changed to red. With the outburst of the plague in 1563, came an attempt to enforce a terrible system of compulsorily shutting up infected families. The doors and windows in such houses were to be closed, and no inmates were to leave the premises and no visitors to be allowed for forty days, No better incubator on a large scale could possibly have been devised for both breeding and intensifying the virulence of the plague bacillus, or whatever may be the contagium vivum of this disease. This compulsory shutting up of the sick with the healthy amounted to a compulsory infection of many of the unfortunate inmates who might THE PLAGUE. 255 otherwise have escaped, and very naturally the order was frequently infringed. In 1568 the Lord Mayor of London drew up instructions for the Aldermen for dealing with the plague. It was enacted that constables and officers should search out infected houses and report to the authorities. In other words, that there should be notification by the police. All infected houses were to be shut up, and no person to be allowed to come out for twenty days. All bedding and clothes used by the victims were to be destroyed. At Westminster these instructions were to be enforced under a penalty of seven days in the stocks, with imprisonment to follow, making in all -a punishment of forty days. In 1581 the Lord Mayor transferred notification from the constables to searchers. Two honest and discreet matrons in every parish were to search the body of every such person that happened to die in the parish. They were ordered to make a true report to the clerk of the parish, and the said clerk had to report to the wardens of the parish. For failing to notify, the penalty was an exemplary term of imprisonment. The searchers were of course likely to be offered heavy bribes by the people to suppress reports, owing to their anxiety to avoid the shutting up of infected houses. The continued prevalence of the plague led to the publication, in 1593, of a book by Simon Kellwaye. One chapter "teacheth what orders magistrates and rulers of citties and towns should cause to be observed," which included among other regulations that no dunghills were to be allowed near the city, and the streets were to be watered and cleansed. No surgeons or barbers who let blood were to cast the same into the streets. All those visiting and attending the sick to carry something in their hand to be known from other people ; and if the infection were in few places, all the people were to be kept in their houses during the time of their visitation, and when this was over, all clothes, bedding, and other such things used upon the sick, were to be burnt. In 1603 Thomas Lodge recommended that discreet and skilful men should be appointed in every parish to notify sickness to the authorities, and so cause them to be visited by expert physicians, and that such as were sick should be separated from the whole, and that isolation hospitals should be built outside the City in separate and unfrequented places. In 1665 the Great Plague of London occurred, and was attributed by some to the importation of an infected bale of silks from the. Levant. . According to Hodges the disease stayed among the common people, and hence was called The Poor's Plague. He criticised the system of shutting up infected houses, and strongly recommended that those who were untouched in infected houses should receive " accommodation outside the city." The sick were to be removed to convenient apartments provided on purpose for them. To quote his own words, " Timely separation of the infected from the well is absolutely necessary to be done." For the purification of houses his directions were to place " a chafing 256 INFECTIVE DISEASES. dish in the middle of the room, where proper things were burnt and exhaled all around." The use of sulphur and quicklime was mentioned.* Preventive measures were drawn up and published by the Lord Mayor and Aldermen. Examiners in Health, watchmen, and searchers w.ere appointed. Surgeons were selected to assist the searchers in making their reports, and a fee of twelve pence was allowed for every case. The disease was immediately notified to the Examiner of Health. Rules for disinfec- tion were made, and every infected house was shut up, and no one removed except to a pest-house or tent. Orders were issued for cleaning and sweep- ing the streets. Hackney coaches were not to be used after conveying patients to the pest-house until they had been well aired. Regulations were also made dealing with loose persons, assemblies, and drinking taverns. The plague was scarcely over before the whole city was in flames. A new city speedily rose upon the ashes of Old London. A few sporadic cases of plague are given in the London Bills of Mortality down to 1679r when they finally ceased. London was sterilised by the great fire. '* Great as this calamity was," wrote Thomas Pennant, "yet it proved the provi- dential cause of putting a stop to one of far more tremendous nature. The plague, which, for a series of ages, had, with very short intervals^ visited our capital in its most dreadful forms, never appeared there again after the rebuilding of the city in a more open and airy manner ; which removed several nuisances, which if not the origin of a plague, was assuredly one great pabulum, when it had seized our streets." In the years 1720-22 there was a terrible outburst of plague in France. It was attributed at Marseilles to importation by a ship from Syria. This caused a panic in England, and the Lords Justices considered it necessary for the public safety that measures should be taken to defend the country from a fresh invasion of this disease. Dr. Richard Mead was entrusted with drawing up the required recommendations. Mead laid it down as an essential doctrine that the plague was not native to this country, and therefore the first thing was to prevent importation, and if such a misfor- tune occurred, it was to be prevented from spreading. How was this to be accomplished ? Briefly stated, his system was as follows : Lazarettoes were to be provided for the reception of infected men and merchandise. The healthy were to change their clothes and to be kept in quarantine, and the sick were to be kept remote from the healthy and their clothes destroyed. If, through a miscarriage in the public care, by the neglect of officers or otherwise, the disease was imported, then " the civil magistrates were to make it as much for the interest of the afflicted families to discover * During outbreaks of the plague amulets were extremely popular. Walnuts filled with mercury, pieces of cloth coated with arsenic, and arsenical cakes, were very generally worn. The College of Physicians recommended issues on the arms and legs. Dr. Hodges wrote, that the more of the ulcers that were made the better, although their largeness answered as well as more in number. If: two issues were preferred, it was recommended to make one on the left arm and the other on the opposite leg. A somewhat similar plan was adopted in Circassia by small-pox inoculators. RELAPSING FEVER. 257 their misfortune, as it was when a house was on fire, to call in the assistance of the neighbourhood." The shutting up of infected houses was condemned in the strongest terms, and a system of notification and isolation was proposed on the lines originally suggested by Dr. Hodges. 1 . .1 Council of Health was to be established, and entrusted with such powers as might enable them to see all their orders executed with im- partial justice. 2. NotificfitioH. — The ignorant old women employed as searchers were to be replaced by understanding and diligent men, who were to report cases immediately to the Council of Health. 3. Isolation. — Physicians were at once to be despatched to visit the suspected cases, and when the suspicion of plague was confirmed, all the families in which the sickness occurred were to be isolated. The sick were to be separated from the sound, and isolation houses to be provided three or four miles out of the town. The removal of the sick was to be made at night, so as to avoid the danger of spreading infection, and all possible care was to be taken to provide such means of conveyance for the sick that they might receive no injury. The poor were to be isolated in houses provided for the purpose, but the rich were to be allowed to be in their own homes provided that care was taken to separate the healthy from the sick, and no pains were to be spared to provide clean and airy apartments. All expenses were to be paid by the public, and a reward was to be given to the person who made the first discovery of infection in any place. Mead further pointed out that general sanitation must be carefully attended to. Officers were to see that the streets were washed and kept clean from filth, carrion, and all manner of nuisances. Beggars and idle persons were to be taken up, and such miserable objects as were fit neither for the hospitals nor for the workhouses, were to be provided for in an establishment for incurables. Houses also were to be kept clean, and sulphur was to be used as a disinfectant. After centuries of experience we have learnt that the necessary conditions for avoiding the plague are more accurate knowledge on the part of the profession and the public of the way in which the disease spreads, and the adoption of sanitary precautions, which must include personal cleanliness, sanitary dwellings, absence of overcrowd- ing, immediate notification, prompt separation of the sick from the healthy, disinfection of infected dwellings, destruction of infected clothing, and extra-mural burial or, better still, cremation. It was because the very reverse of these sanitary conditions existed that the virus of the plague found a suitable environment in Old London and in recent times in Hong- Kong. RELAPSING FEVER. Relapsing or famine fever is a contagious disease producing a slate of high fever lasting about seven days, followed by apparent " 17 258 INFECTIVE DISEASES. recovery, and in about fourteen days by another attack of fever, which may be repeated after another week. Starvation, in association with overcrowding and filth, is intimately connected with the causation of the disease. The subjects of the disease contaminate the air around them, and the virus is principally conveyed by tramps and dirty people. Obermeier discovered spirilla in the blood during the paroxysms of fever. The constant occurrence of the spirillum in relapsing fever, and the fact of its not being found in any other conditions, render it very probable that it is the cause of the disease. D N\ m FIG. 124.— SPIRILLUM OBERMEIERI IN BLOOD OF MONKEY INOCULATED WITH SPIRILLA AFTER REMOVAL OF THE SPLEEN (SOUDAKEWITCH). Spirillum Obermeieri (Spirochceta Obermeieri, Oohn).— Threads similar to the Spirillum plicatile. In length they are mostly 16 to 40 yu-, with regular screw-curves. They move very rapidly, and exhibit peculiar wave-like undulations. They are absent from the blood during the non-febrile intervals, but are found in the interior of leucocytes in the spleen. In blood serum and 50 per cent, salt solution, they preserve their movements. In cover-glass preparations they are readily stained by any of the aniline dyes, and in sections, by preference, with Bismarck brown. They are not TYPHUS FEVER. — YELLOW FEVER. 259 found in the urine, sweat, or saliva. They have not been cultivated artificially on any nutrient media. Monkeys have been successfully inoculated with blood containing the spirilla by Koch, Carter and Soudakewitch ; and Koch found the spirilla in the vessels of the brain, liver, and kidneys, after death. According to Soudakewitch a fatal result is produced in monkeys if the spleen is removed, and the spirilla are found in great numbers in the blood ; but if the spleen is not excised the spirilla rapidly disappear, and recovery follows. Miinch and Motschutkowsky transferred blood containing the spirilla to healthy persons, and produced typical relapsing fever. TYPHUS FEVER. Typhus fever is a highly contagious disease, which lasts for two or three weeks, and produces a measly eruption. Like the plague, it is intimately associated with overcrowding and filth, and is liable to occur where these conditions exist in cities, in armies, and in prisons. The virus produces profound changes in the blood, and after death the internal organs are found to be congested, especially the lungs, which are very friable. The spleen is softened and often enlarged, and the blood is dark and imperfectly coagulated. The virus is dissipated by fresh air. It is given off by the breath of patients, and possibly from the skin. It clings to the clothes of patients, and the disease may be conveyed by their agency. One attack, as a rule, confers immunity. Some persons are naturally insusceptible, failing to contract the disease though daily exposed to it. Hlava has described a bacterium which he believes to be the specific micro-organism. Thoinot and Calmette found the same bacterium with others, but there was no particular micro-organism constantly present. There can be little doubt that the nature of the contagium is unknown. Stamping-out System. — Sanitary precautions, and especially the operation of the Public Health Acts in relation to lodging- houses, prisons, and the better housing of the working classes, have been instrumental in almost completely stamping out the disease in this country. YELLOW FEVER. Yellow fever is a disease of tropical climates, characterised by abdominal tenderness, hsemorrhagic vomiting (black- vomit), and jaundice. The disease may end fatally, or recovery occur in about two or three weeks. It is especially prevalent in the West Indies and in parts of North and South America. 260 INFECTIVE DISEASES. The virus may be conveyed by infected ships, and has in this way made its appearance at British and French seaport towns. The disease is generally believed to be contagious, but the source of the virus is not known. According to Sternberg the virus is not conveyed by water, but spreads where there is overcrowding and filth. Bacteria in Yellow Fever. — Freire asserts that there is a specific micrococcus in yellow fever which ca,n be grown on all ordinary nutrient media, and that cultures can be used for protective inoculation with satisfactory results. Carmonay Yalle also claims to have discovered the contagium ; but Sternberg, who has carried on investigations extending over several years, maintains that there is no characteristic micro-organism present in the blood or in the tissues after death. Aerobic and anaerobic cultures were made from the blood, liver, kidney, urine, stomach, and intestines. The liver was found to contain after death a number of bacilli, most frequently Bacillus coli communis and Bacillus cadaveris. Blood or fresh liver does not produce any disease in rabbits or guinea-pigs, but liver tissue kept for forty-eight hours and inoculated subcutaneously in guinea-pigs is extremely pathogenic. Similar results occur after inoculation of healthy liver which has been kept in the same way. We may conclude from these experiments that the nature of the contagium is unknown. Stamping-out System. — Sternberg states that there are many facts relating to the origin and extension of yellow fever epidemics which support the theory that the virus is present in the evacua- tions, and that accumulations of fsecal matter and of organic material of animal origin furnish in certain climates a suitable soil for the development of the contagium. According to this view the evacuations should be thoroughly disinfected, and with other sanitary precautions and efficient quarantine at seaports, the disease may be stamped out, and the danger of importation from the natural home of the disease reduce:! to a minimum. CHAPTER XIX. SCARLET FEVER. — MEASLES. SCARLET FEVER. SCARLET FEVER is a highly contagious disease peculiar to man. It produces inflammation of the tonsils and adjoining parts, fever, and a general punctiform eruption. The period of incubation is about a week, and the rash usually appears on the second day. In some cases the disease manifests itself in an extremely mild form, known as latent scarlet fever, in which there is only a slight febrile attack, or a mild sore throat, with very little or no rash. Many cases would not be recognisable as such if they were not capable of conveying scarlet fever, or unless other cases followed or occurred simultaneously which were undoubtedly typical cases of the disease. The occurrence of such cases in the early history of an epidemic often causes the greatest difficulty in tracing the origin of the outbreak, and indeed in some cases renders it quite impossible to do so. The virus is given off by the skin, in desquamation, and possibly by the urine. It maintains its vitality in clothing for months, and sometimes longer. It may also be conveyed by the hands of the physician to women during parturition. The disease may be transferred by subcutaneously inoculating persons, who have not previously contracted scarlet fever, with virus obtained by puncturing the eruption on the skin. After death the internal organs appear to the naked eye more or less healthy. The liver is soft, the kidneys are congested, the ileum is inflamed, and Peyer's patches enlarged and congested; but these conditions are also produced by other causes. There are inflam- matory changes in the lymphatic follicles of the tonsils, and the larynx and trachea. Other morbid lesions, especially in the kidneys, are associated with the sequelae and complications, and though commonly occurring in scarlet fever are also found in other diseases. 261 262 INFECTIVE DISEASES. These changes appear to be clue to the poison which is in the blood, and is excreted by the kidneys. The epithelium is in a state of cloudy swelling, a condition found in other febrile diseases and in septic poisoning. Bacteria in Scarlet Fever. — The occurrence of micro-organ- isms in cases of scarlet fever has been observed by several investi- gators— Coze and Feltz, Crooke, Loftier, Babes, Heubner and Bahrdt, and notably by Frankel and Freudenberg, and more recently by Klein, the author, Raskin, and others. Coze and Feltz found cocci in the blood, and Crooke, in cases of scarlet fever with severely affected throat, found bacilli, cocci, and streptococci in the organs of the throat, and cocci in the internal organs. Crooke left it an open question whether these cocci were the specific organisms of scarlet fever, or were to be regarded as diphtheritic or septic associates. He inclined, for clinical reasons, to the latter view. Loffler, in cases of scarlatinal diphtheria, found the same chain - forming micrococcus which he had found in typical diphtheria. Babes was able constantly to prove the presence of a strepto- coccus in inflammatory products secondary to scarlatina. Heubner and Bahrdt, in a fatal case of scarlet fever in a boy, complicated with suppuration of the finger and knee-joints, and with pericarditis, found a streptococcus identical in form with Strepto- coccus pyogenes, but cultivations were not made. The secondary infection started from diphtheritically affected tonsils, which were followed by retro-pharyngeal abscesses. Frankel and Freudenberg examined, for micro-organisms, three cases of scarlatina with well-marked affection of the throat. In all three cases they obtained cultivations of cocci from the submaxillary lymphatic glands, spleen, liver, and kidney. These cocci could not be distinguished from Streptococcus pyogenes derived from pus, nor from the undoubtedly identical streptococcus which one of them (A. Frankel) had repeatedly cultivated in large numbers from puerperal affections. In two of the cases Streptococcus pyogenes was the only organism present, and in all three cases it was far in excess of other colonies which developed. The organisms were also found in sections of the organs by microscopical examination. Frankel and Freudenberg could in no way distinguish the strepto- coccus in scarlatina from the streptococcus in pyaemia and septi- caemia. The identity of this streptococcus with Streptococcus pyogenes and Streptococcus puerperalis was established by comparison of their macroscopical and microscopical appearances in cultivations on SCARLET FEVER. 263 nutrient agar-agar, nutrient gelatine, and in broth, both at the ordinary and at higher temperatures, and also by experiments on animals. They concluded that it could be stated with certainty that the organisms in question did not stand in causal relation to scarlet fever. They considered that special methods of microscopical and biological research were apparently needed for demonstrating the true scarlet fever contagium, which probably was especially present in the skin. They considered that the presence of the a. b. c. FlG. 125.— PURE-CULTIVATIONS OF STREPTOCOCCUS PYOGENES. (a) On the surface of nutrient gelatine ; (b) In the depth of nutrient gelatine : (c) On the surface of nutrient agar. streptococcus was due to a secondary infection, to which the door was opened by the lesions of the throat — a view which was sup- ported by the fact that the organisms were found in submaxillary lymphatic glands. They preferred to use the term " secondary " to " complicated " or " combined " infection, because this expresses the fact that by the effect of the scarlatinal virus the soil is rendered suitable for this ubiquitous microbe when it has once gained an entrance. This streptococcus was found by Klein in five out of eleven cases INFECTIVE DISEASES. of scarlet fever in man, twice in association with certain other micro-organisms, and three times alone. The micro-organisms were isolated by inoculating tubes of nutrient gelatine, solidified obliquely, by streaking the surface with blood taken from the finger, the arm, or the heart after death. Those cases from which the organism was obtained were all cases with ulcerated throat, and the culture experiments, from the living patient, were made on or about the day at which the temperature was at its maximum. Name of Patient. Condition of Tonsils. Source Micro-Organisms Blood. Isolated' Death or Recovery. 1 L F . Severely Finger Streptococcus Ultimately aged 5 ulcerated recovered. fStaphylococcus ] 2 K F , aged 2 Much ulcerated . " pyogenes aureus •( Liquefying micro- coccus 1 Died of f pygemia. ^Streptococcus J 3 4 H — L — , aged 8 (a woman), Ulcerated Much Arm J Staphylococcus (Streptococcus None [•Recovered. [Not stated.] aged 40 ulcerated 5 (a girl), 55 •5 5J aged 19 6 B M , Ulcerated Finger i Streptococcus Recovered. aged 15 7 E W . Much „ None [Not stated.] aged 22 ulcerated 8 R H , Ulcerated • ;» aged 8 9 F G , M Heart Streptococcus Died. aged 2J 10 E F — „ None ,, aged 3 11 E B , Advanced „ | 5J aged 20 months ulceration i Klein regards this streptococcus as the actual cause of scarlet fever in man. The author, Raskin, Holmes, and others who have investigated this subject agree with the conclusions of Fraiikel and Freudenberg. The author is convinced that the streptococci in suppuration, puerperal septicaemia, pyaemia, and septicaemia, and in certain cases of measles, scarlatina, and diphtheria, are identical ; and from overwhelming evidence we are justified in concluding that — (1) The nature of the contagium of scarlet fever is unknown. (2) The streptococcus regarded by Klein as the contagium is the Streptococcus pyogenes. MILK-SCARLATINA. 265 (3) This streptococcus is found, sometimes in company with Staphylo- coccus pyogeiies aureus, as a secondary result in scarlet fever and many other diseases, and its exact relation to scarlet fever and its identity with the streptococcus from pus and puerperal fever, were definitely established in 1885 by Frankel and Freudenberg. MILK- SCARLATINA. It would not be necessary to say anything further on the etiology of scarlet fever if the generally accepted belief, that scarlet fever is a disease peculiar to man, were accepted by the Medical Department of the Local Government Board; but the theory is officially held that scarlet fever is in its origin a disease of cows. Bovine scarlatina is supposed to be an eruptive disease of the teats, and it is maintained that the virus, by contaminating the milk, produces scarlet fever in the human subject. As this theory is very naturally accepted by many medical officers of health, and is mentioned in English medical text-books, it will be necessary to discuss this question in considerable detail, and especially as these opinions were promulgated in this country with official support, and have since been proved to be erroneous. The theory of the origin of the exanthemata in diseases of the lower animals is a very old one. The Arabians imagined that small- pox arose from the camel. Jenner adopted a similar theory, and expressed his belief that small-pox originated in the horse, being generated by horses suffering with " greasy " hocks. Thus Jenner wrote : "May not accidental circumstances have again and again arisen, still working new changes upon it, until it has acquired the contagious and malignant form under which we now com- monly see it, making its devastations among us ? and from a consideration of the change which the infectious matter undergoes from producing a disease in the cow, may we not conceive that many contagious diseases now prevalent amongst us may owe their present appearance, not to a simple, but a compound origin ? For example, is it difficult to imagine that measles, scarlet fever, ulcerated sore throats, and spotted skin, all spring from the same source, assuming some variety in their forms according to the nature of their new combinations?" Baron informs us that tliis idea was prevalent in Jeimer's mind as early as 1787. It is related that in that year he accompanied his nephew, George Jenner, into a stable to look at a horse with diseased heels, and, pointing to them, he it-marked : " There is the source of small-pox. I have much to say 266 INFECTIVE DISEASES. on that subject which I hope in due time to give to the world." And again in 1794, when writing in connection with this subject, he adds : " Domestication of animals has certainly provided a prolific source of diseases among man." Jenner's views were found to be incorrect, and it was shown by Loy and others that the grease bears no relation to cow-pox, and it is now known that Jenner mistook horse-pox for the disease known as the grease. No one at the present day supports Jenner's theory of small-pox in man arising from any disease of the horse. Indeed, the origin of small-pox from a disease of the horse was not upheld even by Jenner's pupil and nephew, Henry Jenner. The latter promulgated the idea that small- pox originated from the cow. He believed that small-pox, in fact, was cow-pox intensified in its virulence by being passed through man. He thus expressed himself • " Nor may it, perhaps, be too hypothetical to suppose that the cow-pox may possibly be the small-pox in its original unadulterated state, before it became contaminated by passing through the impure and scrofulous habits of human constitutions." The theory of the origin, in animals, of human febrile diseases was, later, advocated by Copland, who stated, firstly, that scarlet fever in man was originally a disease of the horse, and that it formerly occurred, and had recently occurred, epidemically as an epizootic among horses ; secondly, that it was communicated in comparatively modern times from horses to man ; thirdly, that it might be, and had been, communicated to the dog. But this opinion has not been accepted, for the disease called scarlatina in the horse is a lion- infectious disease, generally attack- ing but one or two horses in a large stud. • It neither spreads by contagion nor infection ; and Williams states that it is impossible to transmit it from the horse to any other animal, and that many cases of the so-called scarlatina of the horse are in reality identical with purpura. The theory was again revived, but in another form, and has been adopted by the Medical Department of the Local Government Board. Owing to failure in tracing, in some cases of milk scarla- tina, the contamination of the milk from a human source, the theory was started that in such cases the disease is derived from the cow — that, in other words, there is a disease, scarlet fever in the cow, which is responsible for outbreaks of scarlet fever in man. In 1882 an epidemic of scarlatina in St. Giles and St. Pancras was investigated by Mr. W. H. Power for the Board. The disease was distributed with a milk supply from a Surrey farm. In this case MILK-SCARLATINA. 267 two facts were ascertained : the one, that a cow recently come into milk had been suffering from some ailment from the time of her parturition, of which loss of hair in patches was the most con- spicuous manifestation ; the other, that there existed no discoverable means by which the milk could have received infective quality from the human subject. In 1885 an outbreak of scarlet fever occurred in Marylebone in connection with milk from a farm at Hendoii, and again Power failed to establish infection from any human source in any commonly accepted way — such, for example, as handling of milk, or milk utensils, by persons carrying scarlatina infection. But on examining the cows with a view to ascertain any new condition pertaining to them, it came to light during the inquiry that some of them, which had recently been introduced from Derbyshire, were suffering from a vesicular disease of the teats. At this stage Klein became associated with Power in the inquiry; and their belief in the existence of a disease among the cows on the farm capable of producing scarlatina among human consumers of the cow's milk, became unreserved. Klein took away with him samples of milk, contents of vesicles, and discharges from ulcers, and afterwards two of the cows were purchased and kept under observation. Dr. Cameron of Hendon has given a detailed description of the clinical history of this disease. He expressed his belief that it was a specific disease capable of being communicated to healthy cows by direct inoculation of the teats with virus conveyed by the milker from a diseased animal. The condition of the teats is described as follows : The teats became enlarged, swollen to nearly twice their natural size, and cedematotis. On handling them there was no feeling of induration. Vesicles appeared on the swollen teats and upon the udder between or near the teats. These varied in number from two to four on a teat, and in size from a pea to a horsebean. The vesicle contained a clear fluid. The vesicles were rubbed and broken in milking, and left raw sores, sometimes red, in other cases pale in colour, with raised, ulcerated edges. Sometimes a few accessory vesicles formed around the margin of these ulcerated sores. After the rupture of the vesicle a brown scab formed, which might remain attached for five or six weeks, or fall off in ten days or a fortnight, a smaller one forming ; afterwards. A thin, watery fluid exuded from un.lrr the scab, and the sore ultimately healed. Cameron examined the teats of several cows five or six weeks 268 INFECTIVE DISEASES. after they were attacked. The scabs then varied in size from a shilling to a florin ; they were about one-eighth of an inch thick in the centre, thinning off towards the edges. Some of the cows were also suffering from an eruption on the rump and hind quarter, consisting of patches of eczematous crusts. When a crust was picked off, the hair came off with it, exposing a raw, moist sore, the crusts and sores looking exactly like eczematous scabs and sores ; but this condition corresponds in description with eczema, the result of ringworm which is very common in young stock. In addition to his own observations, Cameron obtained infor- mation from the farmers, and others familiar with cows, who thought they recognised in the disease at the farm one stage of a disease which they were able to describe. Cameron thus gives an account of what he and his informants together would regard as a connected clinical history of the disease. He did not see the earlier symptoms, and hence these were of necessity learnt from other persons. The account, therefore, of these symptoms was to be held liable to future correction or modification. Cameron stated that he learnt this disease was capable of being communicated to milkers by inoculation with virus from the vesicles on the teats, though the milkers on the Hen don farm escaped. " A trusty informant received the virus into a recent scratch on the forefinger while milking a diseased cow. General weakness, malaise, and loss of appetite resulted, and after about four or five days a vesicle or small blister appeared on the finger. This broke, and several others formed on the back of the hand. The whole hand and fingers became swollen and inflamed, the inflammation extending in broad lines as far as the elbow. The general disturbance lasted a fortnight." In the course of the inquiry, Cameron adds that it was strongly asserted by several people, who examined the cows, that they were suffering from cow-pox. He, however, dismissed the diagnosis of cow-pox on the ground that no papule had been observed or subsequent formation of pustule, areola, or pitting, and because the vesicles were not umbilicated. These reasons given for dismissing the diagnosis of cow-pox at Hendon were totally inadequate ; a comparison having been made between the characters of the eruption of vaccinia as it appears on an infant's^arm, instead of the eruption of the natural or so-called spontaneous disease 011 the teats of the cow. MILK-SCARLATINA. Klein stated that on the teats and udders of two cows which he investigated there were several flat irregular ulcers, varying in diameter from one-quarter to three-quarters of an inch. Some ulcers were more or less circular, others extended in a longitudinal direction on the teat. The ulcers were covered with a brownish or reddish-brown scab. The animals looked thin, but not strikingly so. In feeding capacity, milking power, arid body temperature there was nothing abnormal. Four calves were inoculated in the corium of the groin and the inside of the ear, with scrapings from the ulcers after removal of the crusts. In one, which may be taken as an example of the result obtained, there was vesiculation at the margin of the spot inoculated, and in the centre the commencement of the formation of a crust. On the seventh day each sore on the ear had enlarged to about half an inch in breadth, and was covered in its whole extent by a brownish crust. On the eighteenth day they had all healed up and become converted into flat scars. To search for micro-organisms, Klein removed the crust from an ulcer on the teat, scraped off the most superficial layer, squeezed the ulcer, and made cover-glass preparations. Tubes of nutrient gelatine and nutrient agar-agar were also inoculated, and a streptococcus was isolated which in morphological and cultural characters agreed with those of Streptococcus pyogeues. Two calves were inoculated in the groin with the cultivated micro-organism. One calf died in twenty-seven days. At the necropsy there were found peritonitis, and hsemorrhagic spots on the omentum ; the liver, kidneys, and lungs were congested, and there were petechise under the pleura, and pericarditis. The second calf was killed, and at the necropsy the lungs and kidneys were congested, and there were hsemorrhagic patches on the spleen. In these cases, the post-mortem appearances and anatomical features recalled to Klein the lesions of scarlatina. In the kidney, for example, the cortex was congested, and there were haemorrhages, glomerulo-nephritis, and granular or opaque swelling of the epithelial cells and infiltration with round cells. From the blood of the heart the streptococcus, which had been used in the inoculation, was recovered. In view of this evidence it was con- cluded that the streptococcus was the virus of the cow disease, and that it produced in calves a disease very closely resembling that of scarlatina in man. Two of the cows selected from the Hendon farm were killed, and it was observed in one that the lungs were congested, and 270 INFECTIVE DISEASES. that there were numerous adhesions by recent soft lymph between the lower lobes of the lung and the costal pleura. In the liver there were several reddish streaks and patches. The spleen and kidneys, with the exception of slight congestion, appeared normal. Sections of the kidney showed well-marked glomerulo- nephritis and infiltration of the sheath of the cortical arterioles with numerous round cells. The epithelium of the convoluted tubules was swollen, opaque, and in many places disintegrating. In the other cow there was great congestion of the lungs and pleural adhesions ; the cortex of the kidney was congested, but its medulla was pale. On microscopic examination there was a good deal of round - celled infiltration in the walls of the inf undibula and bronchi in the lung, and round the arterioles in the kidney. In sections of the ulcers on the teats, the corium was found to be infiltrated throughout the whole extent of the ulcer with round cells. In the superficial layers of the stratum Malpighi, close to the stratum lucidum, as also in the stratum lucidum itself, there were numerous cavities of different sizes. These cavities lay side by side, the most superficial ones being covered by the stratum lucidum, or extending between the layers of this stratum. At the marginal parts the cavities, although placed side by side, were well separated from one another by thicker or thinner trabeculse, composed of epithelium, while at or near the centre of the ulcer these trabecula? were destroyed, the cavities had become confluent, and the covering layers of the cuticle having here also given way, their contents extended on to the free surface of the ulcer. In short, Klein states that all the anatomical details of the distribution and arrangement of these cavities recalled vividly the conditions observed in the vesicles of cow-pox. Yet as a result of this investigation he concluded that the cow disease at Heridon was bovine scarlatina, and that towards its study and supervision every effort ought to be directed in order to check the spread of scarlet fever in man. As a result of this conclusion, the Board of Agriculture resolved to have the whole subject fully investigated, and the author was directed to study the bacteriology and micro-pathology of this disease and to report thereon. Professor Axe investigated the origin of the outbreak of the disease in the cows, and Professor M'Fadyean carried out an investigation into the possibility of inoculating cows with the virus from cases of scarlet fever in man. MILK-SCARLATINA. 271 THE AUTHOR'S INVESTIGATION. An outbreak of an eruptive 'disease of the teats, alleged to be identical with the so-called Hendon cow disease, was raging in some farms in Wiltshire. In this case every facility was given by the owner of the estate for a thorough investigation into the disease. Not only were animals sent from the farm to London, but the author was allowed to visit the farms, to inspect all the infected animals, and to make every investigation, with the hearty co- operation of the bailiff of the farms, and the voluntary assistance of the head cowmen and those under them. Some of these cowmen were unusually intelligent, while two had had experience of cows for more than half a century. Thus, there was not only every opportunity for studying the disease on the lines indicated by Klein, but it was possible by repeated visits to the farms to enter into the clinical history of the disease in the cow, to study very fully the nature of the disease on the hands of the milkers, and to trace the probable mode of its introduction on the estate, and the way in which it spread from one part of the herd to another. Two cows were sent to London with disease of the teats and of the udder between the teats. On the right teats of one there were numerous sores, covered with crusts varying in size and in thickness, and generally fissured. In some they were flat, in others conical ; some were with difficulty removed with forceps, others were readily detached. The crusts varied in colour from reddish-brown to very dark brown or almost black. On detaching or scraping a crust there was a granulating and somewhat indurated base. On the right anterior teat there were several ulcers, from which appa- rently the thick crusts had been detached, and new scabs were forming. On the left posterior teat there were unusually large, dark brown, or blackish crusts, covering a very extensive are;i of ulceration, extending over the whole of the lower third of the teat. In the other cow from Wiltshire there was the same disease on the teats, but not in such a severe form. The sores were covered with thick crusts, but though varying in size they were more regular in form, and more circumscribed. Having entirely removed the crusts from some of the ulcers, a number of inoculations in nutrient gelatine and nutrient agar-agar were made from the discharge, and cover-glass preparations were made and stained in the ordinary way. Cultures were obtained of the organisms commonly found in pus. 272 INFECTIVE DISEASES. With the discharge and with scrapings from the ulcers two calves were inoculated. Of the two calves, one was inoculated by scarification in both ears ; the other, a small calf, was scarified in the left ear. Scrapings from the ulcers were rubbed into the places thus prepared. In addition, in the small calf an incision was made through the corium in the left groin, scrapings from different ulcers on the teats were well rubbed in with the blade of the scalpel, and a portion of crust inserted into a small pocket in the subcutaneous tissue. In the ears and the groin there were positive results. In the large brown calf one of two places inoculated in the right ear passed through the following changes : On the third day there was apparent vesiculation and commencing formation of crust. From day to day the crust thickened, and on the eighth day the crust was at its height and detached at its edges. By removing the scab an ulcer was exposed ; there was slight inflammatory thickening. About the thirteenth day the ulcer had quite healed. Very similar appearances resulted in the ear of the smaller calf. The result of inoculation in the groin was of a very much severer character. In the course of two or three days the incision had apparently commenced to heal by scabbing, but there was a surround- ing area which was inflamed, and painful on manipulation. The inflammatory thickening which resulted continued to increase around the seat of inoculation, and the thickening could be felt to extend deeply into the groin. Suppuration followed, and on firm pressure pus welled up through the wound. The wound then showed very little disposition to heal, and the calf began to exhibit marked constitutional symptoms. During the second week after inoculation the animal became very dull, and was reported by the attendant as refusing to feed. Diarrhoea supervened, and lasted for several days, and bloody urine was passed. The calf was also noticed to cough, and the cough gradually increased in severity. Thirty-six days after the date of inoculation it was decided to kill the calf and examine the condition of the viscera. The appearances which were found at the post-mortem examination were as follows : — The upper and middle lobes of each lung were adherent to the walls of the chest ; there was congestion, especially of the middle lobe, and patches of recent adherent lymph. Posterior parts of the upper lobes of both lungs were completely consolidated, and on section varied in colour from brick-red to greyish -white. The interlobular tissue was infiltrated with inflammatory products, which mapped out the tissue of the lung in small indurated areas, in which the tissue was granular-looking and friable. THE AUTHOR'S INVESTIGATION. 273 These appearances in the upper lobes were due to septic pleuro-pneumonia. They closely resembled, and were supposed to be due to, infectious pleuro-pneumonia. They were, however, found identical with the con- dition observed in septic pleuro-pneumonia in calves, and the disease was not conveyed by infection to other animals in the same stall. Scattered through the other lobes of both lungs were white, mostly firm, nodules raised above the level of the surface of the lung. They were surrounded by a zone of congestion, and in some cases sections were composed of indurated, in others of friable, lung tissue. In the posterior part of the right upper lobe there was a recent infarct. The bronchial glands at the roots of each lung were enlarged to two or three times their natural size, and were firm and hard on section. The parietal surface of the pericardium was covered with recent adherent lymph. The visceral surface of the pericardium was normal. Along the external surface of the aorta were chains of enlarged lymphatic glands connected by dilated lymphatic vessels. These glands were dark red or purplish in colour, from haemor- rhage into their substance. The heart was normal, and the endocardium not stained. There were chains of red glands on the oesophagus similar to those along the aorta. The appearance of the mesenteric glands was very striking. The mesentery, along the lymphatic vessels, was dotted with glands, varying in size from a large shot to a pea, which were deep red or prune-coloured. In addition, there were here and there enlarged glands without haemorrhage into their substance, and greyish in colour. There were scattered petechiae on the spleen. The kidneys were firm on section, and there was marked congestion in both, while it was more pronounced in one kidney than the other. The liver was congested, the congestion being more marked in patches. Sections from the consolidated upper lobes showed under the micro- scope thickening of the pleura and infiltration with round cells. The exudation filled the alveoli, and was breaking down in some cases in the centre. The vessels were injected, and there were haemorrhages into the alveoli. The periphery of the lobules was infiltrated with round cells. In sections of the kidney there was slight infiltration around glomeruli and arterioles with round cells ; the epithelium in the convoluted tubules was granular and disintegrating ; there was haemorrhage in the straight tubules, and engorgement of vessels. In sections of liver the inter- and intra-lobular vessels were engorged ; there were interlobular collections of round cells displacing the liver cells, and the interlobular connective tissue was infiltrated with round cells ; the liver cells were granular and cloudy. There can be no doubt from the symptoms and post-mortein appearances that this calf had been suffering from septicaemia .is the result of introducing the septic virus and crust subcutaneously in the groin. The two Wiltshire cows were killed, and there was nothing of importance to note in one, but in the other an incision into the udder revealed an enormous abscess. 18 274 INFECTIVE DISEASES. Though the naked-eye appearances of the kidney in this case were practically healthy, the results of examining sections of the kidney under the microscope were extremely instructive and interest- ing, as they showed that marked changes had taken place which were indicative of septic complication. The sections showed glomerulo- nephritis ; there was infiltration of the capsule of Bowman with round cells ; there was infiltration also of the sheaths of the vessels with round cells, especially in the cortex. The blood-vessels in the boundary zone of the medulla were engorged, the arterioles of the glomeruli were also engorged, and there were slight haemorrhages into the capsule. The epithelium of the convoluted tubules was granular, opaque, and in some parts breaking down. Sections of the ulcers of the teats of these cows were also carefully examined, and the appearances corresponded exactly with the description given by Klein. On visiting the farms it was found that there were altogether about a hundred and sixty cows. Only a few had proved refractory, and had not taken the disease at all. The rest had contracted the disease in varying degrees of severity. About fifty at a time were dry, and they escaped until they were in milk again. The milk was drunk on the farms and in the village, and a quantity was supplied to a large town. Most careful inquiries were instituted to ascertain the existence of scarlatina among consumers of the milk. So far the research was completely analogous to the Hendon investigation; but, in spite of the contamination of the milk, no cases of scarlatina were found either on the farms or in the village, and there was no epidemic in the town in which the milk was distributed. The disease, in fact, was cow-pox, and in no way connected with scarlet fever ; and to assist others who may undertake a similar inquiry the details will now be given of the author's investigation into the nature of the outbreak in Wiltshire. THE DISEASE PROVED TO BE COW-POX. Locality of the Wiltshire Outbreak. — There is considerable interest attached to the fact that the farms were situated a few miles from Cricklade. They are close to the borders of Gloucestershire, and about twenty-five miles from Berkeley. They are, therefore, within that district in which in Jenner's time cow-pox was particularly prevalent. Time of Year. — The outbreak commenced about the end of September 1886, and lasted until about the middle of December. In an outbreak AN OUTBREAK OF COW-POX. 275 in 1885, a few miles from these farms, but on a separate estate, the disease appeared in June and July. Origin of the Outbreak. — The author made careful inquiries as to the origin of the outbreak, but beyond ascertaining with certainty that the disease appeared first at one farm, and was conveyed from this to the other farms, all evidence was negative. The milkers were unable to say whether it commenced in one particular cow or whether it broke out in several simultaneously. The only information which could be obtained, which was very suggestive, was that the milkers were in the habit of receiving their friends from neighbouring farms on Sundays. The friends would assist in the milking, to get the work done as quickly as possible on these occasions. As it was reported that the same disease had occurred that summer on a neighbouring farm, it is quite possible that it was introduced by one of the milkers' friends. Mode of Dissemination. — When the disease made its first appearance, the bailiff, attributing it to the farm being, for some reason, unhealthy, decided to remove the cows to other farms. The herd was therefore divided and sent to two other farms. From these cows the disease was communicated to healthy cows, and, as this interchange was repeated, not only of the cows, but of the milkers, the disease was communicated to four separate farms. In all cases the disease was limited to the teats, and was conveyed from the teats of a diseased cow to the teats of a healthy cow by the hand of the milker. In no case was there any evidence of the disease being produced in healthy cows by other means than contact. Bulls and dry cows remained free from the disease, while the cows in milk, numbering about a hundred and twenty, were all attacked, with the exception of about a dozen, which proved to be entirely refractory. These facts explain how it is that the disease has been known from time immemorial as the "cow-pox." We never hear of cattle-pox or bull-pox. We have not, in other words, to deal with an infectious disease like cattle-plague or pleuro-pneumonia, but with a disease which is communicated solely by contact. The disease was only observed in the cows in milk, and was limited to the parts which come in contact with the hand of the milker. The virus was mechanically transferred from diseased to healthy cows, being communicated to all, or nearly all, the animals in the same shed, whether the milker had vesicles on his hand or not. Character of the Erujrtion on the Cow. — In a recent case which was carefully examined the teats were visibly inflamed, partly red and partly livid in colour. On each teat there were vesicles, some broken, and others which appeared to be just forming. In other cases there was nothing more than the remains of broken and dried vesicles, and more or less characteristic crusts on the teats. On visiting a byre at the time that the cows were brought in to be milked, it was a striking sight to look along the line and see one animal after another affected with the eruption ; and thus one character 276 INFECTIVE DISEASES. of the disease was clearly shown — the tendency to spread through a whole herd. On examining the eruption carefully, the degree of severity was found to differ very much in different animals. In a few cases the condition was most distressing, both to the cow and to the observer. In such cases the teats were encrusted with huge, dark brown or black crusts, which, when handled in milking, were broken and detached, exposing a bleeding, suppurating, ulcerated base. Such ulcers varied in size from a shilling to a florin, and in form were circular, ovoid, or irregular. Weeks afterwards, when the animals had recovered, the site of these ulcers was marked by irregular scars. All the milkers agreed as to the general characters of the malady, laying particular stress on the teats being red, swollen, and painful when handled. Vesicles would then appear on the teats — two, three, four, or more on each teat. They were soon broken in milking, and irritated into sores, which became covered with thick crusts. From four to six weeks elapsed before they had entirely healed. Other more observant milkers insisted that before the teats were red and swollen, spots or pimples first appeared which came to a head. This head increased if it was not broken, which might be the case if it was situated between the bases of the teats, until it formed a greyish vesicle of the size of a threepenny-piece or even larger. General Symptoms in the Cow. — As to the general condition of the cows nothing abnormal was observed. They appeared in the best of health, and in only one particular was any difference from their condition in health stated to exist. This was, that in the majority of the cases there could be no doubt that the milk had diminished. This might escape notice by inexperienced milkers in any particular animal, but the total amount of milk supplied by the herd was considerably below the average. History of the Eruption communicated to the Milkers. — The most striking characteristic of this outbreak was the communicability of the disease to the milkers. A milker, with vesicles which presented typically the characters of casual cow-pox, was taken to London and kept under observation. The various cases will be described in the order in which they first presented themselves, their history being given as much as possible in their own words. CASE I. — J. R., milker, informed the author that he was the first to catch the eruption from the cows. He stated that it came as a hard, painful spot, which formed " matter " and then a " big scab." He had been inoculated about seven weeks previously. He pointed to the scar which remained on his right hand. This scar presented the characters of an irregular cicatrix, indicating considerable loss of substance. He stated that he had also two places on his back, where he supposes he had inoculated himself by scratching. He had continued milking ever since, but had had no fresh places. CASE II.— W. H., milker. He stated that he was inoculated from the cows about the same time as J. R. They were the two milkers of the herd in which the cow-pox first made its appearance. The eruption AN OUTBREAK OF GOW-POX. 277 appeared in one place on each hand. He pointed to two irregular scars as the remains of the eruption. CASE III.— J. L., milker, stated that he also caught the disease from the cows. On his right hand a spot appeared which formed a blister, then discharged matter and produced a bad sore. Lumps formed at the bend of his elbow and in his armpit. He lost his appetite, felt very poorly, and was obliged to leave off work for two or three days. CASE IV. — W. K., a labourer on the farm, was put on as a milker to take the place of one of the others with bad hands. After his fifth or sixth milking — that is to say, about three days after first milking the cows, —pimples appeared on his hands, which became blistered and then ran on to bad sores. He pointed to three irregular scars on the first and third fingers and palm of the right hand. Lumps appeared in his elbow and in his armpit, but he did not feel very poorly in consequence. CASE V.— J. F., milker, stated that about a month ago he noticed spots which appeared on both hands. His fingers swelled and were pain- ful. He said it came first like a pimple, and felt bard. Then it " weeped out" water in four or five days. There were red marks creeping up to his arm. There was a sort of throbbing pain, and he could not sleep at night. On the right hand there was a scar, but on the left hand there was an ulcer about the size of a shilling covered with a thick black crust. The crust was partially detached, and exposed a granulating ulcer. It was in this stage the exact counterpart of the ulcers on the cow's teats. CASE VI.— W. H., junior, milker, stated that he had both hands bad about a month previously : first on the index finger of the left hand, and then on the right hand on his knuckle and between the first and second fingers. He said that it came up like a hard pimple, and the finger became swollen and red. After a few days it " weeped out " water, and then matter came away. Both his arms were swollen, bat his left arm was the worst. About a fortnight after, he noticed kernels in his armpits, which were painful and kept him awake at night. His arms became worse, he could not raise them, and he had to give up milking. He also had had a " bad place " on the lower lip. On examina- tion, I found that the axillary glands were still enlarged and tender. He volunteered the statement that the places were just like the sore teats. CASE VII. — J. H., the bailiff's son, also milked the cows. He had a sore on the upper lid of his right eye and on his left hand. In both cases he had been previously scratched by a cat, and the scratches were inoculated from the cow's teats. The right hand also had been inoculated. The eruption broke out a fortnight previously. His hands were swollen, red, and hot. He felt very poorly and went to bed. Little spots like white blisters appeared on the back of his right hand. His mother remarked that they "rose up exactly as in vaccination." Thick dark brown scabs formed. He was very ill for two or three days, but did not send for a doctor. He had painful lumps at the bend of his arm and in the armpit. He gave up milking, and had not taken to it since. On examining him, the thick crusts on his right hand were identical (UNIVERSITY ^^r^ Pir irm 278 INFECTIVE DISEASES. with the stage of scabbing in vaccinia. The scabs fell off in about three weeks to a month, and left permanent, depressed scars. CASE VIII. — W. P., milker. This case was pointed out on the occasion of another visit, and is the only one in which the eruption was seen in its earlier stages. The history of this boy is as follows. He had taken the place of one of the other milkers who had vesicles on his fingers, and had been obliged to give up milking. After the seventh time of milking he noticed a small pimple on his right cheek. This became larger and vesicular. On examination it presented a depressed vesicle with a small central yellowish crust and a tumid margin, the whole being surrounded by a well-marked areola and considerable surrounding induration. On raising the central incrustation a crater-like excavation was seen, in which lymph welled up and trickled down the boy's cheek. On the following day the crust had re-formed, and was studded with coagulated lymph. The areola became more marked, and on pricking the margin of the vesicle, the exuding contents were slightly turbid. From this day the surrounding infiltration increased enormously, the whole cheek was inflamed, and the eyelids so oedematous that the eye was almost closed. There was enlargement of the neighbouring lymphatic glands. The crust which had re-formed thickened day by day. It retained the character of central depression, and was situated on a reddened, raised, and indurated base (Plate VII.). From this date the surrounding induration gradually diminished. The crust changed in colour from dark brown to black, and finally fell off, leaving an irregular, depressed scar. This scar, when seen several months afterwards, was found to be a permanent disfigurement. The eruption appeared on the fourth day after exposure to infection, and allowing two days for incubation, the vesicle was at its height on the seventh or eighth day, and a typical tamarind- stone crust fell off on the twenty-first day after infection, leaving a depressed, irregular cicatrix. A vesicle also formed on the thumb of the left hand. Two days after the pimple appeared on his cheek, the lad said that he first noticed a pimple on his thumb, and this, on examination, presented a greyish flattened vesicle, about the size of a sixpence. Later, its vesicular character was much more marked, and a little central crust had com- menced to form. The margins became very tumid, giving it a marked appearance of central depression. The vesicle was punctured at its margin with a clean needle, and from the beads of lymph which exuded a number of capillary tubes were filled. Two days afterwards suppuration had commenced, the vesicle con- tained a turbid fluid, and the areola was well marked. Later, the crust had assumed a peculiar slate- coloured hue, and, on pressing it, pus welled up through a central fissure. The areola had increased, and there was considerable inflammatory thickening. The lymphatic glands in the armpit were enlarged and painful. Though there was deep ulceration, which left a permanent scar, the ulceration did not assume DESCRIPTION OF PLATE VII. Casual Cow-pox. FIG. 1. — Case of W. P , a milker, infected from the teats of a cow with natural cow-pox. There was a large depressed vesicle with a small central crust and a tumid margin, the whole being surrounded by a well-marked areola and considerable surrounding induration. FIG. 2. — The same case a week later, showing a reddish-brown crust on a reddened elevated and indurated base. Plate TOT. Eg 2. CASUAL COW-POX. OF THB UNIVERSITY OF r AN OUTBREAK OF COW-POX. 279 quite so severe a character as in some of the other milkers. Possibly this may be accounted for to some extent by the fact that the pock was covered with a simple dressing instead of being subjected to the irritation and injury incidental to working on the farm. Re-vaccination of the Milker*. — There were in all eight milkers, varying in age from seventeen to fifty-five, who had vesicles on their hands from milking the cows. Seven had been vaccinated in infancy, but not since ; one had been revaccinated on entering the navy at fifteen. They were all revaccinated by a public vaccinator after complete recovery from the casual cow-pox (that is to say, from three to four months afterwards), and were all completely protected. On the other hand, two of the three milkers who had escaped infection from the casual cow-pox were also vaccinated, with the result in one of typical revaccination, in the other of very considerable local irritation. Retro-vaccination of Calves. — The result of retro-vaccinating calves with the humanised lymph was strictly in accordance with the experience of Ceely, who has pointed out that in retro-vaccination from the milker's hands the results are doubtful, and depend greatly on the animals selected. "Those of a light colour and with thin skins were generally preferred, but often without avail, scarcely one-half of the operations succeeding." " Vaccine lymph, in passing from the cow to man, undergoes a change which renders it less acceptable and less energetic on being returned to many individuals of the class producing it ; some refuse it altogether." Two cases out of four succeeded, and an eruption was produced with all the typical characters of vaccinia, but running rather a rapid course, and the protection passing off after a few weeks, while the result obtained in calves inoculated with pus or scrapers from ulcers was in accordance with what is well known to occur if pus instead of lymph is taken for carrying on calf to calf vaccination. That the cow-pox in Wiltshire was identical with the so-called Hendon cow-disease there can be little room for doubt, for in both cases we find that — 1. The disease spread through a whole herd of milch cows. 2. The disease was characterised by the appearance of vesicles, which were broken by the hand of the milker, and irritated into deep ulcerations. 3. The disease was conveyed from one cow to another by the hand of the milker. 4. The vesicular eruption was communicable to the hand of the milker. 5. The disease was not fatal, and in cows which were killed and examined the post-mortem appearances could not be distinguished from accidental complications. 6. The naked-eye appearances and the duration of the ulcers of the teats were the same. 280 INFECTIVE DISEASES. 7. Sections of the ulcers showed under the microscope identical appearances of a cellular character, and the purulent discharge of the ulcers contained pyogenic cocci. 8. The results produced by inoculation of calves with the septic virus were identical. If we examine the chain of argument which has been brought forward to maintain the existence of cow-scarlatina at Hendon, we find that it was urged : — 1. That the Hendon cow disease was a disease in which the post-mortem appearances resembled scarlatina. 2. That this disease was associated with a streptococcus, which produced, by inoculation in calves, a disease with post-mortem appearances similar to those of the Hendon cows. 3. That a streptococcus regarded as identical with the one above mentioned was found in certain cases of scarlatina in man, which when inoculated in calves produced post-mortem appearances similar to the post-mortem appearances in the original Hendon cows and in certain cases of scarlatina in man. But the microscopical appearances of the kidney of a Wiltshire cow were identical with those which were regarded as indicating scarlatina in a Hendon cow ; and, indeed, the statements as to the post-mortem appearances in the Hendon cows, when studied, not only do not necessarily indicate scarlatina, but they cannot even be considered of primary importance, or as throwing much light on the question of scarlatina at all. The description of the naked- eye appearances in both cows only suggests coincident pleurisy or pleurisy with pneumonia. The microscopical appearances in both were suggestive of septic complication. A careful examination of the post-mortem appearances of calves inoculated with scraping of an ulcer of a Hendon cow, or with cultivations of the streptococcus from certain cases of scarlatina, brings to light much more striking changes. These appearances, however, cannot be regarded as indicative of scarlatina. They are in reality the post-mortem appearances of septic poisoning, and occur commonly in many diseases. This is clearly shown by com- paring the post-n.ortem appearances in the calf which was killed while suffering from septicaemia as the result of inoculation from the ulcers of a Wiltshire cow. These visceral changes are not to be distinguished from the post-mortem appearances described in the calves inoculated by Klein, Consequently, that the strepto- AN OUTBREAK OF COW-POX. 281 coccus found in certain cases of scarlet fever should produce on inoculation in calves certain post-mortem appearances which are found in many diseases, and should fail to produce fever, ulceration of the tonsils, or scarlatinal rash, or any condition in the least resembling, clinically, the disease in man, and yet that the result should be regarded as scarlatina in the calf, is a conclusion quite untenable. It is true that visceral lesions similar in character were produced in calves whether inoculated with scrapings or with streptococci from ulcers of the Hendon cows or with streptococci from certain cases of scarlet fever. In both cases the streptococcus is pathogenic, and inoculation of Streptococcus pyogenes or the inoculation of septic virus, is liable to produce septicaemia, These facts constitute a mass of evidence which justifies the conviction that the pathological data which appeared to support the theory that the vesicular disease of the teats of cows at Hendon was scarlatina in the cow, admit of an entirely different interpretation^ and there can be no longer any doubt that the milk was not infected by the cows but with the virus of scarlet fever from some human source which Mr. Power failed to discover. All the other evidence reported to the Board of Agriculture pointed to the same conclusion. The disease at Hendon was admittedly introduced from Derbyshire ; and from Professor Axe's report it appears that only a part of the herd was sold to the farmer at Hendon; other cows with the same eruption were transferred to other dairy farms, and the disease communicated to healthy cows as at Hendon, but in no instance did scarlet fever occur among the consumers of the milk. At the farm of the brother of the dealer the disease was communicated to three of the milkers, and the eruption diagnosed by Dr. Bates as vaccinia. All this evidence must be regarded as conclusive. The con- tamination of the milk at Hendon with scarlet fever must neces- sarily have been a mere coincidence ; and the conclusion that the milk could not possibly have become infected from any human source is untenable. Professor Axe even ascertained that scarlet fever existed at Hendon during several months of 1885, and that the dwellings where cases occurred stood within MX hundred yards of the cowsheds which contained the incriminated cows, and that out of fourteen men on the farm six lived in a district where cases occurred. Professor Axe has also stated that the father and brother of a girl with scarlet fever, visited the dairy during her illness. Whether any of those engaged on the 282 INFECTIVE DISEASES. farm suffered from latent scarlet fever does not appear to have been ascertained. There is, it is true, no evidence to show that any one daily carried infection to the milk, but the exact path of infection is not always easy to trace ; and because it was not actually traced it was hardly reasonable to assume that the possibility of contamination from a human source could be altogether eliminated. In attempting to communicate scarlet fever to cows Professor M'Fadyean confirmed the negative results which had been experi- enced in some earlier experiments by Klein. In 1882 Klein inoculated and fed cows and yearling heifers with diseased products from human patients, using desquamated cuticle and the discharges from the throat ; but the experiments all failed. M'Fadyean's failures were still more marked. Cows and calves were inoculated with blood from scarlet fever patients, and they were made to drink water thickened with desquamated cuticle, but all the experiments proved unsuccessful. The author believes that the outbreak at Hendoii was one of cow-pox, which was prevalent in this country in 1886. The outbreak in Wiltshire could not be distinguished bacteriologically or clinically or in its micropathology, from the disease at Hendon, and the Wiltshire outbreak proved on investigation to be true cow- pox. This conclusion was questioned at the time, as cow-pox was generally believed to be extinct in England ; but that view is entirely fallacious, and the author's conclusions have since been fully confirmed by independent observers, whose work will be referred to in another chapter (p. 321). Stamping-out System. — The Notification Act of 1890 may be voluntarily adopted in sanitary districts, but it would be a great advantage if notification were carried out uniformly all over the country. Prompt information may lead to detecting the origin of cases of scarlet fever, and isolation and disinfection will assist in pre- venting its spread. Epidemics have occurred on a large scale owing to scarlet fever existing among those engaged in dairy work, and the precaution not being taken of stopping the milk supplied to the consumers. Scarlet fever cannot be so readily controlled as small- pox, for it may be spread by mild cases before the nature of the disease is suspected, ancl small-pox cannot be conveyed in milk. MEASLES. 283 MEASLES. Measles is a contagious disease peculiar to man. It lasts for one or two weeks, and produces fever, catarrh of the respiratory mucous membrane, and a characteristic rash. It is highly contagious, especially before the nature of the disease is revealed; there is consequently great difficulty in preventing its spread in schools and households. The contagium appears to be given off from the body, principally if not entirely, by the breath. One attack is pro- tective against future attacks. The whole population of a country may acquire a certain degree of immunity. Measles introduced into countries where it was previously unknown assumes a most malig- nant form. There are no characteristic post-mortem appearances. Bacteria in Measles. — Micrococci have been found in the blood, catarrhal exudation, and skin, by Keating, Babes, and others, but they are accidental epiphytes of no importance, or associated with secondary complications, as in scarlet fever. Canon and Pielicke have found in the blood small bacilli varying in form. They do not grow on nutrient agar or blood serum, but cultures were obtained by pricking the finger of a patient suffering from measles, and allowing the blood to drop into sterilised broth. After a few days the broth became cloudy, and later, a flocculent deposit formed. The bacilli were also obtained from the nasal and conjunctival secretions. The nature of the contagium of measles is unknown. Stamping- out System. — Measles is not easily controlled by the stamping-out system ; it is, in fact, extremely difficult, almost impos- sible, to prevent its spread, as it is especially infectious during the period of incubation. Notification, isolation, and disinfection assist in controlling an epidemic, but the value of the system does not apply to the same extent in measles as in other infectious diseases. CHAPTER XX. SMALL-POX. — CATTLE PLAGUE. SMALL-POX. SMALL-POX is an infections and inoculable disease of man, charac- terised by sadden and severe fever, followed in forty-eight hours by a characteristic papular eruption which gradually becomes vesicular and then pustular. The virus is contained in the vesicles, and in a concentrated form in mature pustules. It also passes into the air from the breath and skin. Infection may occur from the dead body, and clothes and bedding may retain the contagium for months. One attack, as a rule, gives immunity against future attacks. Small-pox is undoubtedly a disease foreign to this country. Its home is in the East. Some of the old writers held that it spread to Europe from Alexandria about the year 640 A.D., following in the wake of the Arab conquests in Egypt, Palestine, Persia, along the Asiatic coast, through Lycia, Gallicia, along the coast of Africa, and across the Mediterranean to Spain ; others maintained that it was not introduced until the end of the eleventh or beginning of the twelfth century, by the returning Crusaders. At any rate, small- pox was imported from the East, and probably from Egypt. Hero- dotus, who visited Egypt, leads us to infer that epidemics were unknown there during the rule of the Pharaohs ; but Egypt undoubtedly became a hotbed of pestilence during the Mohammedan occupation. Prosper Alpinus imagined that both the plague and the small-pox were concocted in the putrid waters of the Nile, but he would probably have been more correct if he had suggested that they arose from the insanitary condition of the Arab conquerors and their filthy camp followers, who did their best to destroy all that remained of that magnificent civilisation which had existed in the days of the ancient Egyptians. We do not know the exact period at which small-pox was first imported into England, and the records of the disease are very meagre until the sixteenth century. 284 SMALL-POX. 285 In 1593 Simon Kell \vaye appended to his work on the Plague a short treatise on the small-pox. " Oftentimes," he wrote, " those that are infected with the plague are in the end -of the disease sometimes troubled with the small pocks or measels, as also by good observation it hath been seen that they are fore-runners or warnings of the plague to come." According to Kellwaye the disease arose from the " excrements of all the foul humours in our bodies, which striving with the purest doth cause a supernatural heat and ebullition of our blood, always beginning with a feaver in the most part." Small-pox steadily increased in the seventeenth century until it was a formidable scourge, for no advantage was taken of all the experi- ence which had been gained in dealing with the plague. No public measures were adopted to cope with the disease, and the people came to regard the new pestilence as a visitation which was unavoidable. Early in the eighteenth century, small-pox inoculation was introduced, and this was superseded in the nineteenth century by vaccination. Examination of small -pox cases after death does not reveal any characteristic lesions in the internal organs, but sections of small- pox vesicles show an important structure. A vesicle is formed by the exudation raising up the outer layer of epidermis, and the chief feature is the formation of a vacuolated structure in which, especially in the later stages, bacteria are found in abundance. Bacteria in Small-pox. — Cohn and Weigert found cocci in variolous lymph. Hlava found Streptococcus pyogenes in the pustules, and Garre streptococci in the internal organs in a case of variola hsemorrhagica. In a fatal case of variola complicated with pemphigus Garre found a streptococcus in the pemphigus vesicles. Klein and Copeman have found a small bacillus which they regard as characteristic, but its biological characters are unknown, as it will not grow on any nutrient media. The bacteria commonly found in variolous pus are the usual pyogenic organisms. The nature of the contagium of small-pox is unknown. Protective Inoculation. — Experience had taught that a person was not, as a rule, attacked with small-pox a second time; but when and how the method of artificially inducing a mild form -of the disease was discovered, or when this preventive treatment was first employed, is unknown. Avicenna of Bokhara was credited with the discovery, and it was supposed that the practice was carried by Tartar and Chinese traders to Surat, Bengal, and China, and by the Mahommedan pilgrims to Mecca. In Constantinople it was supposed by some to have been introduced from the Morea by an old woman, and by others by the women of Circassia. The 286 INFECTIVE DISEASES. Circassian women fastened three needles together, and pricked the skin over the pit of the stomach and heart, the navel, the right wrist, and the left ankle. The variolous matter was applied to the bleeding points, and the eruption came out in five or six days. In Constantinople scarifications were made on the forehead, wrists, and legs, and carefully selected virus applied to the incisions. The needle used was a three-edged surgeon's needle, or the operation was performed with a lancet. The virus was obtained by pricking the vesicles, and pressing out the matter into a clean glass vessel. The Armenians preferred to be inoculated in both thighs. In Barbary a slight wound was made between the thumb and forefinger, and the virus obtained from a mild form of small-pox. In Hindustan the operation was performed at certain seasons of the year, and a preparatory regimen enforced. The inoculators were very careful in the selection of the virus, as they had learnt its varying intensity, and they were credited with being able to control the amount of the eruption. They preferred to inoculate the outside of the arm, midway between the wrist and the elbow in males, and between the elbow and the shoulder in females. The skin over the part to be inoculated was first well rubbed with a piece of cloth ; then, with slight touches of a small instrument, little wounds were made over an area which might be covered by a small coin, and sufficient to cause just an appearance of blood. A pledget of cotton -wool charged with the variolous matter, and moistened with water, was applied to the wound. This virus was obtained from inoculated pustules of the preceding year. In China the contents of the variolous pustules were dried and kept for several years. If the virus was to be used from fresh pustules the " acrimony " of the matter was corrected by steaming. The dried powder was made into a paste, which was wrapped up in cotton -wool and introduced into the nostril. The Greeks were more cautious in their procedure, and were said to inoculate tens of thousands without an accident. They operated only upon those in perfect health, punctures were made with needles, and the virus was used in the crude state, freshly obtained from the " kindly " pustules of a young child. They were particularly careful in the choice of the "ferment." Dr. Perrot Williams, in 1722, wrote that the practice of com- municating small-pox had long been employed in South Wales. The oldest inhabitants said that it had been a common practice with them "time out of mind," but Lady Mary Wortley Montagu was responsible for the general adoption of small-pox inoculation in SMALL-POX. 287 England by persuading physicians in London to employ it. Lady Mary had her child inoculated in Turkey. An old Greek woman inocu- lated one arm, and Mr. Maitland, surgeon to the Embassy, the other. The disease ensued in due course with an eruption of a hundred pustules. This was the first time that the Byzantine method of inoculation was performed upon an English subject. In 1721 Dr. Harris delivered a lecture before the College of Physicians, and described the successful inoculation of four children of the French consul at Aleppo, by means of a thread imbued with variolous pus. A "daughter of Lady Mary was inoculated in England by Maitland in 1721, and subsequently a number of criminals were inoculated by him. Incisions were made through the cutis, and pledgets which had been steeped in variolous pus from ripe pustules, were applied to the wound. This was known as Maitland's or the reformed operation, but it was soon modified, as troublesome ulcers resulted. Shortly afterwards Maitland encountered another obstacle. The child of a Mr. Batt was inoculated, had plenty of pustules, and soon recovered, but six of Mr. Batt's domestic servants, " who all in turn were wont to hug this child while under this operation, and whilst the pustules were out, never suspecting them to be infectious, were all seized at once with the right natural small-pox of several and very different kinds." Dr. Jurin in 1729 reverted to the' Eastern method, and recom- mended virus from a mild case of small-pox, but the virus was still taken from perfectly maturated pustules, and the operation continued to be followed by bad results. In order to diminish the risks, Burgess in 1766 advocated certain improvements. An incision about an inch long was made on each arm through the cuticle, but not so deep as to wound the cellular tissue. A variolous thread was laid along the whole length of the wound and fixed with plaster. Ulcerations and other accidents continued to take place, and a new epoch in the history of inoculation was the introduction of the Suttonian method, in 1764-6. It was said that Mr. Button, with his assistants, inoculated one hundred thousand persons. The method was kept secret at first, but the essential points were all discovered and published by Dr. Dimsdale. Dimsdale recommended a very slight puncture with a lancet wet with variolous matter. Subsequently, Sutton published an account of his method, and the result of his operation may be given in his own words. "The lancet being charged with the smallest perceivable quantity (and the smaller the better) of unripe, crude, or watery matter, immediately 288 INFECTIVE DISEASES. introduce it by puncture, obliquely, between the scarf and true skin, barely sufficient to draw blood, and not deeper than the sixteenth part of an inch. Neither patting, nor daubing of the matter, in or over the punctured part, is at all necessary to its efficacy. This practice, indeed, is rather prejudicial than otherwise, as it may affect the form of the incision, and thus be apt to confound our judgment upon it. " Indications of the Incision. — In the incipient state of variolous increase in the incision, a small florid spot appears on the part of access, resembling a flea-bite in size ; and on passing the finger lightly over it a hardness is felt not larger than a small pin's head. This florid appearance and hardness denote that the variolous principle is effectually imbibed, and their indications point no farther, unless the progress to vesication be very slow, in which case an uncomfortable number of pustules may be expected to follow. The florid spot in most instances of inoculation is somewhat larger, or more extended on the second, than on the third day after the insertion. "About the fourth day from inoculation, should the incision begin to vesicate, an itching sensation will be complained of on the place of insertion — the occurrence of which symptom is the first indication of a favourable event, yet not of sufficient importance to justify any present relaxation in the preparatory proceedings. " The vesication of the incision in most instances will begin to be visible on the fourth or fifth day after the insertion of the matter ; the sooner it becomes so, the more favourable may be expected to be the event. The extent or diameter of the vesication at this stage does not usually exceed that of a large pin's head, and it has invariably a dint or small depression." Adams obtained still more striking results by inoculating with variolous lymph from pearl-pox, a mild variety of small-pox. Starting with lymph obtained from this benign form of small-pox, and selecting the cases, and carrying on arm to arm variolation, the results obtained were practically identical with the phenomena obtained by inoculation of the arm with cow-pox lymph. Similar results were obtained by Guillou, but more rapidly. In 1827 there was an epidemic of variola, and Guillou, having no vaccine virus, took variolous lymph from a girl fifteen years of age on the fifth clay of the eruption. The case was one of varioloid or mild small-pox, attri- buted to previous vaccination. The variolous lymph was inserted in ten places on the arm of a healthy infant still at the breast. This inoculation produced ten beautiful " vaccine " vesicles, from which, on the ninth day, forty -two infants were inoculated under the eyes of two of the local authorities. These furnished virus for the inoculation of one hundred, who were inoculated in the presence of magistrates and many medical men. This experiment was repeated with success. Variolous lymph was taken from two lads at school, and in ten SMALL-POX. 289 cases produced appearances with a perfect similarity to ordinary vaccination. Thiele produced a benign vesicle in the following manner. Variolous lymph was diluted with warm cow's milk, and inoculated like ordinary vaccine lymph. Large vesicles resulted. There were febrile symptoms from the third to the fourth day, and a secondary onset of fever much more pronounced between the eleventh and fourteenth days. The areola was strongly marked, and not con- fined to the inoculated place, which was occasionally surrounded by minute secondary vesicles. After watching through ten removes, the vesicles finally assumed the characters of an ordinary vaccination with cow-pox lymph. As soon as the secondary fever ceased to occur inoculation was practised from arm to arm without diluting the lymph with cow's milk. The lymph was designated lacto-varioline, and the result was variolation in its mildest form. The result of variolating the cow will be discussed in another chapter. Small-pox inoculation, or variolation, protected the individual when genuine small-pox was produced, and .endangered the com- munity. Persons inoculated became centres of infection, and con- veyed the disease to others. Haygarth, although in favour of inoculation, strongly condemned its use without precautions to prevent the spread of the disease. " The most serious and solid objection," he wrote, " that has been advanced against inoculation is deduced from a comparison of the Bills of Mortality for a series of years in various places. They show that a larger proportion of inhabitants have died of the small-pox in towns where it is prac- tised than in the same before it was known, or in others where it is prohibited." Even Dr. Dimsdale, an ardent inoculator, admitted that more lives were lost in London than before inoculation commenced, and the practice was more detrimental than beneficial to society ; and he added : " The disease by general inoculation throughout London spreads by visitors, strangers, servants, washerwomen, doctors, and inoculators, by means of hackney coaches in which the sick are sent out to take the air, or by sound persons approaching them in the streets. The poor in London are miserably lodged ; their habitations are in close alleys, courts, lanes, and old dirty houses ; they are often in want of necessaries, even of bedding. The fathers and mothers are employed constantly in laborious occupation abroad, and cannot attend the inoculated sick." In 1798 Jenner, who had practised small-pox inoculation, proposed the use of a benign non-infectious lymph obtained from a disease of the cow or horse as a substitute 19 290 INFECTIVE DISEASES. for variolous lymph, and in 1840 small-pox inoculation was prohibited by Act of Parliament. Stamping-out System. — The disappointing and dangerous results of small-pox inoculation led to a widespread demand for some new method for dealing with small-pox. This induced Haygarth to turn his attention to the subject, arid towards the end of the eighteenth century to bring before the medical profession and the public a plan for stamping out the disease. Haygarth, who was a close observer and an able physician, studied the question of the communicability of the disease from one person to another, and its conveyance by infected clothing and other means, and ultimately drew up rules and regulations for its prevention, the importance of which we are only now beginning to fully acknowledge. Haygarth's essential doctrine was " that mankind was not necessarily subject to the small-pox, and that it was always caught by infection from a patient or the poisonous matter," and might be avoided by observing his Rules of Prevention. These rules comprised a regular system of notification and isolation. Inspectors were to be provided to report cases of small-pox, and people were to be rewarded for carrying out the instructions. Several examples were given of the results at Chester, where the plan was adopted. Haygarth met with considerable encouragement from some of the leaders of the profession. Dr. Fothergill wrote to him in 1778, saying, " I have mentioned the intention of freeing this country from the small-pox to divers of the faculty, and shall continue to do so as it falls in my way. The proposal is variously received, but in exact proportion to their humanity." In 1793 Haygarth made considerable addition to his rules, and urged that legislation should follow to make them compulsory. Provision was to be made to reward the poor for observing the rules, and public thanks to the wealthy for their support were to be published in the parish church and newspapers. Transgression of the rules was to be punished by a fine of from £10 to £50, one half to go to the informer and the other half to the fund which supplied the expense of rewards to the poor, and all details were to be supplied to the press. It was further suggested that Great Britain should be divided iuto districts, including a certain number of parishes or townships, and that to each of them a surgeon or apothecary should be appointed as inspector to see that the regulations were exactly observed. In addition, there were to be directors of inspectors, superin- tended by a commission of Physicians in London and in Edinburgh. All salaries were to be paid by the county rates, and the rewards for observing the rules of prevention were to be guaranteed out of the parish funds. On the requisition of the director and inspector of a circuit, power was to be given to two or more justices of the peace to appoint a separate house for the reception of patients with the small-pox. In conclusion, SMALL- POX. 291 Haygarth maintained that the plague had been completely exterminated from this country, for above a century, by civil regulations, and that there seemed to be little doubt that the small-pox was propagated on principles similar to the plague, and that it also might certainly be exterminated from this island. Haygarth's teachings had a profound influence upon both the profession and the educated public, but his system of compulsory notification was never carried out, for no legislation followed to enforce his recommenda- tions. This is a matter deeply to be regretted, for towards the end of the eighteenth century small-pox was declining in London, general sanitation was making rapid advances, small-pox inoculation, which created fresh centres of infection, was falling into disfavour, small-pox hospitals were built, which served to limit centres of infection, and the profession and the public were influenced by the teaching of Haygarth with regard to the various ways of avoiding the spread of the disease. It only required the compulsory adoption of Haygarth's system uniformly all over the country to have kept the disease in control, if not to have entirely extirpated it from Great Britain. That a similar conviction existed at the time is evidenced by an article which appeared in 1779 in the Medical and Chirurgical Review, in which the following statements were made : — k< Plans for the extirpation of the small-pox have been suggested. . . . To do this, however, the exertions of the physician are incompetent unless they be aided by the powerful hand of Governments, but this has hitherto been withheld. The grand means, however, of extirpating this destructive malady is an early and strict separation of the infected from those that are sound." Small-pox in the present century has been largely controlled by legislation, especially in recent years, by the Public Health Acts for England and Wales, for Scotland, and for Ireland ; the Epidemic and other Diseases Prevention Act ; the Public Health Amendment Act ; the Labouring Classes' Dwellings Acts ; the Housing of the Working Classes Act ; the Public Health (Ships) Act ; the Local Government Board Act — and various orders and memoranda of the Local Govern- ment Board ; the Infectious Diseases Notification Act ; the Infectious Diseases Prevention Act ; and the Public Health (London) Act. By the Public Health Act of 1875 England was divided into Urban and Rural Sanitary Districts, and powers were given to enforce regulations of the Local Government Board for guarding against the spreading of infectious diseases ; to provide medical aid and accommodation for infected persons, to promote cleansing, ventilation, and disinfection, to provide hospitals, to provide for 292 INFECTIVE DISEASES. destruction or disinfection of infected bedding, clothing, and other articles, and to appoint Medical Officers of Health. As to the value of notification and isolation in cities such as London we have the evidence of the Metropolitan Asylums Board. In their Report for 1889 we read in reference to the diminution of small-pox : " These very satisfactory results confirm the view taken by the Committee two years ago to the effect that the rapid and systematic removal from crowded districts of infected persons, each of whom might have become a centre of contagion, is an important factor in stamping out small-pox from the metropolitan population. The notification of cases will also greatly facilitate the action of the managers in this direction." More recently there has been a most striking confirmation of these statements. An outbreak of small-pox occurred in Maryle- bone, and by the energy of the officials of the Board this outbreak was suppressed in a few days by means of notification and immediate isolation. The Isolation Hospitals Act of 1893 gives power to County Councils to provide, or cause to be provided, an isolation hospital in any district within their county. An application to a County Council for the establishment of an isolation hospital may be made by any one or more of the authorities defined as local authorities having jurisdiction in the county or any part of the county. Further, the County Council may direct an inquiry to be made by two medical officers of health in the county as to the necessity of an isolation hospital being established for the use of the inhabitants of any particular district in the county, and in the event of such medical officers reporting that such a hospital ought to be established for the use of the inhabitants of a district, may take the same proceedings in all respects for the establishment of such hospital, as if a petition had been presented by a local authority for the establishment of an isolation hospital for the district named in the report of such medical officers of health. Lastly, the Local Government Act of 1894 provides for the formation of District Councils ; and the powers, duties, and liabilities are principally those which were conferred by the Public Health Act of 1875. In the opinion of the author the Government of this country should enter into friendly negotiations with the Governments of other countries, so that there might be concerted action to prevent an avoidable disease like small-pox. Much good might result from the formation of a permanent International Board of Health. If civilisation is not yet CATTLE PLAGUE. 293 sufficiently advanced to admit of a system of international notification, our Consular authorities should be instructed to give immediate notifica tion of the existence of small-pox in other countries, and every measure should be enforced to diminish the possibilities of importation. The duties of a Central Health Office, presided over by a Minister of Health, should include the collection of information as to the existence of small- pox in other countries, and details should be published in the Annual Reports of the Department. Regulations, for example, for dealing with the importation of rags from small-pox stricken places should be enforced, as in the case of cholera ; and if, in spite of these precautions, isolated cases occurred in this country, they should be dealt with promptly. Notification shopld be enforced uniformly all over the country, and there is not the slightest reason why the authorities and the public should not immediately receive information of the existence of small-pox, whilst to procure immediate isolation we have only to imitate the excellent ambulance system of the Metropolitan Asylums Board. To procure prompt notification there must be no loophole for evading the Act, and there should be a heavy penalty for failure to notify not only small-pox, but any caxe which may reasonably be supposed to be one of small-pox. The police should be required to report any case of small-pox in common lodging-houses or shelters ; they should have power to require any tramp suffering from small-pox, or from any disease which may reasonably be supposed to be small-pox, to be examined by the medical officer of the Union, and kept under observation, or transferred at once to the isolation hospital ; and inmates of the workhouse should be daily inspected, and no case -allowed to leave when there is the least suspicion of small-pox infection. Objections no doubt will be raised to this proposal, but the frequency with which small-pox is spread by tramps fully justifies these measures. All these measures should be carried out as a matter of routine, and without the semblance of panic. Isolation should be uniformly enforced all over the country, and vaccination should be relegated to the position of a voluntary auxiliary measure, which should never be allowed to take the place of sanitary regulations to stamp out the disease. CATTLE PLAGUE. Cattle plague is a highly contagious disease of bo vines producing high fever, and characterised by an eruption with a resemblance to human small-pox. The disease is transmissible to other ruminants, and is inoculable in man. One attack gives immunity against future attacks. Cattle plague and small-pox are not intercom- municate, and are specifically distinct diseases, but the resemblance between them was recognised from early times. Ramazzini published an account of the cattle pest in Italy in 1711, and described the pustules which broke out over the body as similar to those of variola in 294 INFECTIVE DISEASES. kind and appearance. Dr. Layard, in 1780, described this disease of horned cattle as an eruptive fever of the variolous kind, with the appearance and stages of small-pox. This resemblance was endorsed by Murchison, one of the Commissioners appointed in 1866 to inquire into the origin and nature of cattle plague. Murchison pointed out that in both diseases the eruption con- sisted of pustules and scabs, and that in both it extended from the skin to the interior of the mouth and nostrils ; in both, the pustules and scabs were preceded or accompanied by patches of roseola ; in both, they were occasionally interspersed with petechise ; and in both, they sometimes left behind pitted scars and discolora- tions on the cutis. The other prominent symptoms of rinderpest were also those of small-pox — viz., pyrexia, lumbar pain, salivation, and running from the nostrils, alvine flux, albuminuria, hsematuria, and " the typhoid state." The anatomical lesions of the internal organs in rinderpest and unmodified small-pox were identical — viz., congestion or inflammation of the mucous membranes of the air passages and digestive canal, patches of ecchymosis and even gangrene of the stomach and other mucous surfaces, and darkly coloured blood. In both rinderpest and small-pox the duration of the pyrexial stage was on an average about eight days. In both diseases a peculiarly offensive odour was exhaled from the body before and after death. The two diseases resembled one another in their extreme contagiousness, and in the facility with which the poison was transmitted by foinites. Both diseases were easily propagated by inoculation, and in both cases the inoculated disease was milder and less fatal than that resulting from infection. In both diseases there was a period of incubation, which is shorter when the poison has been introduced by inoculation than when it has been received by infection. Ceely described the result of an accidental inoculation of cattle- plague virus in the human subject. A vesicle was produced which so closely corresponded with the result of inoculated cow-pox that Ceely inclined to the belief that cattle plague was a malignant form of cow-pox. The following is the account of this case as reported by Ceely. Mr. Hancock, a veterinary inspector at Uxbridge, was engaged in superintending the autopsy of a bullock recently dead of cattle plague. His assistant, who was performing the operation, while occupied in removing the skin from the scrotum, accidentally punctured the back of Mr. Hancock's hand with the point of the knife. The puncture being slight was disregarded at the time, but was washed as soon as practicable, and thought of no more. Five CATTLE PLAGUE. 295 days afterwards, a small, slightly elevated, hard pimple was felt and seen on the site of the puncture. This gradually advanced till the ninth day of the puncture, the fourth from papulatipn, when the enlargement became distinctly vesicular. At that time there were but slight constitutional symptoms. On the next day, the tenth from the receipt of the puncture, the fifth from papula tion, and the second from vesiculation, Mr. Hancock consulted Mr. Rayner, of Uxbridge, who, on seeing the hand, inquired if the patient had been handling the udder of a cow, as he thought he could recognise a cow-pox vesicle of the ninth day. The vesicle was distended with thin lymph, its margin elevated and slightly brown, its centre depressed and brownish, and the whole surrounded with a large bright red areola. There was then considerable tumefaction extend- ing from the knuckles to above the wrist. The absorbent vessels were considerably inflamed, and, like the axillary glands, were tender and painful ; the pulse, naturally slow, was accelerated ; there was much pain in the back and limbs, severe distracting headache, etc. ; all of which symptoms continued to increase during the two following days. At the end of that time the diffused areola had extended as far as the elbow. Fifteen days after the puncture, and ten days after papulation, the local inflammation and constitutional symptoms had partially subsided. The vesicle contained a rather turbid brownish fluid, and there were present all the indications of a declining vaccine vesicle. Murchison also saw and described the case, and gave practically the same account of it. He pointed out that the appearances and the entire history were very different from the results of a poisoned wound, but coincided with the appearances seen after vaccination. In 1832 Macpherson, in Bengal, inoculated eleven native children with cattle- plague crusts. There was no result in six, others suffered from local inflammation, and in one a vesicle formed. With lymph from this vesicle other children were inoculated. The results in all were similar in appearance to those of vaccination. Two children were subsequently inoculated with human variola, and were said to be protected. In 1834 Macpherson's example was followed by Mr. Furiiell in Assam. Furnell inoculated four children with cattle-plague crusts without result, but his assistant succeeded with crusts taken from the back and abdomen of the diseased cattle, and carried on the lymph from child to child. In one case there was a general eruption. Furnell inoculated his own child from one of the native children : a copious eruption followed, and the child died. Furnell 296 INFECTIVE DISEASES. after this misfortune issued a strong warning against taking the virus from the cow. The experiments were made in the belief that cattle plague was really small-pox in cattle, and that the virus would protect against human variola. Similar results were obtained by Mr. Wood at Gowalpara in 1838. Bacteria in Cattle Plague. — Semner cultivated streptococci from the blood and lymphatic glands of a sheep suffering from cattle plague. A calf inoculated with a cultivation died in seven days. The cocci were stated to lose their virulence by cultivation, and the weakened cultivation to protect against the virulent disease. The micro-organism was very probably Streptococcus pyogenes, and the calf may have died of septic infection. There can be 110 doubt that the nature of the contagium of cattle plague is unknown. Protective Inoculation. — In the great epidemic of cattle plague in England in 1866, owing to a belief that the analogy between cattle plague and small-pox was closer than it really is, vaccination with cow-pox was attempted as a preventive measure, but was proved to be absolutely useless. Stamping-out System. — When cattle plague was imported in 1865 into London, dairymen and stock-owners made no attempts to prevent the extension of the disease, so that it spread rapidly all over the country through disposal of infected cattle. The losses were enormous, and an Order in Council was passed in July 1865, directing dairymen and others to notify outbreaks of any contagious or infectious disease among the animals under their charge. A Veterinary Department of the State was formed, and inspectors appointed in various parts of the country. A short Act was passed in February 1866. A stamping-out system, consisting of compulsory notification and the slaughter of diseased animals, was soon brought to the notice of the public. There was violent opposition, but nevertheless, after some delay, the system was carried out. The number of cases of cattle plague had reached 18,000 weekly, and on the introduction of the stamping-out system the disease rapidly declined. The disease was again imported into Great Britain in 1872, and there were outbreaks in 1877. In each instance the disease was promptly stamped out, and ever since that year the disease has been kept out of this country. CHAPTER XXI. SHEEP-POX. — FOOT-AND-MOUTH DISEASE. SHEEP-POX. SHEEP-POX, or variola ovina, is an acute febrile disease accompanied by a general vesiculo-pustular eruption, highly infectious, and capable of being propagated by inoculation or clavelisation. It is a common disease in some parts of Europe. In France the disease is called la clavelee, and in Italy vaccuolo. It has been introduced on several occasions into this country, but has been effectually stamped out. As in human small-pox, there are varieties — the benign and the malignant, the discrete and the confluent ; and one attack is protective against the disease in future. It is very closely analogous to human small-pox. Vaccination with cow-pox lymph has been employed to protect sheep from sheep- pox, but unsuccessfully, and lymph for vaccination has been raised from sheep-pox to protect human beings from small-pox. These experiments were first performed in Italy. Marchelli, in 1802, took lymph from the vesicles of sheep-pox, and inoculated children. Sacco repeated these experiments, and found there was no appreciable difference from the results obtained with cow-pox lymph. Dr. Legni carried on the inoculations with ovine virus from arm to arm for several years, and when small-pox occurred in Pesaro, it was said that all those who were inoculated with the sheep virus were protected. Inoculation of children with ovine virus, direct from sheep, was repeated by Sacco and Magnani in 1806. Marson in England succeeded in producing on the human subject a vesicle with the physical characters of the vaccine vesicle. The vesicle had a bluer tinge, and subsequent inoculation of the patient with human variola was ineffectual. Other experimenters were unsuccessful, but their failures, as in the case of variolation of the cow, do not invalidate the results of those who were successful. 297 298 INFECTIVE DISEASES. Sheep-pox and cow-pox are quite distinct diseases. Sheep-pox is highly infectious, whereas cow-pox is only conveyed by direct inoculation, and is never infectious, and further, cow-pox inoculated in sheep does not produce sheep- pox. Bacteria in Sheep-pox. — Hallier and Zurn, Klein, and others, have found micrococci and bacteria in the lymph of the vesicles of sheep-pox, but they are only accidental epiphytes. The nature of the contagium is unknown. Protective Inoculation. — Extensive experiments were carried out in England to test the protective power of vaccination against sheep-pox. According to Marson and Simmonds, it was very difficult to get cow-pox to take on sheep, and when an effect was produced, the resulting affection, even when developed to its fullest extent, was very unlike the same disease in the human subject. In the sheep it seldom produced anything more than a small papule, which occa- sionally resulted in the formation of a minute vesicle, or more commonly, a pustule, which was sometimes, although very rarely, surrounded by a slight areola. Generally, however, neither vesica- tion nor pustulation followed, but a small scab was produced, which soon fell from the site of the puncture, leaving no trace behind. The disease passed quickly and irregularly through its several stages, and terminated by the eighth or ninth day, and not unfrequently even before that time. Lymph was but rarely obtainable, and then only in the smallest quantity, and this on the fifth or sixth day suc- ceeding the vaccination. The effects were only local, and the animal's health was not impaired. Sheep were found to be just as susceptible of the cow-pox virus on subsequent repetition of the inoculation as they were in the first instance, and hence the conclusion that cow-pox was utterly worthless as a protective against sheep-pox. According to Depaulr however, cow-pox takes characteristically on sheep, and sheep- pox lymph inoculated on cows produces a result indistinguishable from the appearances obtained with the inoculation of cow-pox lymph, It is impossible to say whether these conflicting results depended upon the employment in the experiments of different breeds of sheep or different stocks of vaccine lymph. The objection to clavelisation or ovination is that the disease may be introduced in localities where it was previously unknown. By ovination, as in the analogous case of variolation, fresh centres of infection are created, whereas every precaution should be taken to prevent the introduction of the disease. Stamping-out System. — Sheep-pox has been imported into this FOOT-AND-MOUTH DISEASE. 299 country on several occasions. It was introduced in 1847, and again in 1862; in 1865 it was introduced again, and active measures of repression were at once taken. The diseased flocks were carefully isolated, and day by day as fresh cases occurred the diseased animals were killed and buried. Owing to the adoption of these precaution- ary measures, the affection did not extend beyond the flock among which it first appeared. It was introduced again in 1866 at Long Buckby, in Northamptonshire. In this case the disease was exter- minated by the slaughter and burial of the whole flock, and imme- diate application of disinfectants to the hurdles and other things with which the sheep had been in contact. Then it was introduced again in Cheshire, and strict isolation being enforced the infection died out. Since 1866 we have had no outbreak of sheep-pox in this kingdom, but foreign sheep have been landed with sheep-pox in 1868, 1869, 1870, 1871, 1875, 1876, 1878, and 1880, but the disease has been prevented from spreading. The Sheep-pox Order of 1895 provides for the notification of the disease, for disinfection and for compulsory slaughter of infected sheep, and prohibits the movement of diseased or suspected sheep, and the local authority may, if they think fit, order the slaughter of suspected sheep and of sheep which have been in contact with diseased sheep. FOOT-AND-MOUTH DISEASE. Foot-and-mouth disease is a highly contagious and infectious febrile disease, characterised by a vesicular eruption affecting the lips, tongue, roof of the mouth, and feet of sheep, cattle, and pigs, and according to some observers it also attacks horses, poultry, hares, and rabbits. Sometimes the mouth only is affected, in other cases the principal seat of the eruption is in the feet. The vesicles soon break and give rise to ulcers. When these occur in the mouth they cause pain and difficulty in taking food. Extensive ulceration may occur on the feet, causing great pain and lameness. In milch cows it sometimes happens that the eruption occurs on the udder and teats, and it is this manifestation of the disease which has received so much attention from Eayer. The milk is contaminated by the discharge of the vesicles, and is unfit for use, either as food for the human being or for the lower animals. It induces a vesicular eruption in the mouth, larynx, pharynx, and intestinal canal. It acts most vigorously when administered warm to young animals, and calves occasionally die quite suddenly after sucking cows 300 INFECTIVE DISEASES. affected with the eruption on the teats. Fatal effects also result when the milk is administered to young pigs. It has been stated that no injurious consequences arise from the consumption of the milk by human beings, but there is abundant evidence to the contrary, and the conflicting opinions probably arise from the fact that milk is seldom drunk direct from the cow, and rarely in an undiluted form. Hertwig experimented upon himself with milk freshly drawn from a cow with the eruption. He drank a pint, and two days afterwards experienced slight fever, restless- ness, and headache. The mouth was dry and hot, and there was tingling in the skin of the hands and fingers. These symptoms continued for seven days after taking the milk. On the ninth day vesicles had formed on the tongue, principally on the edges, and 011 the mucous membrane of the cheeks and lips (the largest being about the size of a lentil). They were yellowish- white in colour, and contained a whitish turbid liquid, which flowed when the vesicles were pricked with a needle. At the same time a number of vesicles- developed on the hands arid fingers ; and most of them at the time of their first appearance were the size of a millet seed. They were firm to the touch, yellowish -white, and occasioned a slight tingling. The vesicles of the mouth increased in size and eventually broke, and the epithelium detached itself completely from the affected parts, leaving dark red spots, which disappeared gradually. The slight fever present during the first days ceased after the appearance of the eruption ; but from this time, until the disappearance of the red spots, Hertwig felt a continual burning pain in the mouth, and speaking and deglutition caused considerable uneasiness. On the lips the vesicles dried up, and were covered with thin brownish crusts, which fell off ten days after the appearance of the first vesicles. The vesicles which developed on the hands ran a slower course. From the tenth to the thirteenth day they filled with a liquid, like turbid lymph. They were large and confluent, and finally broke and dried up. Bacteria in Foot-and-mouth Disease.— Klein in 1885 isolated from the vesicles a streptococcus which in its microscopical and its cultural characters on gelatine, agar and blood serum resembled Streptococcus pyogeiies. Minute differences in the size of the colonies and in their rate of growth, and in the character of the chains, were observed on making comparative cultures with Streptococcus pyogenes from a human source, but no comparison was made with Streptococcus pyogenes from acute suppuration in cattle. Baumgarten regarded this micro-organism as Streptococcus FOOT-AND-MOUTH DISEASE. 301 pyogenes, and not as the contagium of the disease. The author has pointed out the variation which exists in the size of the chains and of the colonies, and the difference which is found in the rate of growth of cultures of Streptococcus pyogenes, and these variations are especially marked in Streptococcus pyogenes bo vis. Klein believes that the administrations of broth cultures produced the disease in sheep, but the results were very probably due to accidental infection. It is well known how very readily foot-and-mouth disease is spread. The appearance of a case in a flock of sheep or a herd of cattle will be almost certain to be followed by all or nearly all of the other animals being infected with great rapidity. The virus clings to the clothes of shepherds and others who have been in contact with infected sheep, and may be readily conveyed to healthy animals by those who have been visiting infected premises. Schottelius described chains composed of rounded elements, some of which resembled an amoeba or plasmodium. The chains were said to be motile, and delicate growths were obtained in blood serum and agar, and in broth and on potato. Inoculation in sheep and pigs and numerous small animals gave negative results. These organisms were described as streptocytes, to distinguish them from bacteria. Piani and Fiorentini investigated the contents of the vesicles, and also described corpuscular elements exhibiting amoeboid movements. They regarded these bodies as protozoa, and concluded that foot-and- mouth disease is due to their presence. Until a micro-organism is cultivated which will produce sheep- pox in sheep on a farm or on premises where the disease does not exist, and where there can be no possibility of accidental infection, we are fully justified in concluding that the nature of the contagium of this disease is unknown. Stamping-out System. — Foot-and-mouth disease was imported into this country in 1839. It has been successfully dealt with by the stamping-out system, which in this case is very difficult to apply because of the very short period of incubation, and the value of the stamping-out method very greatly depends upon the length of the incubation period. Foot-and-mouth disease very often, from infection to recovery, does not exceed ten days; yet accord in.-: to the reports of the Board of Agriculture, when foot-and-mouth disease exists in a manageable state, perfect isolation and effectual disinfection have proved equal to the complete control of the spreading of the infection, and the final extinction of the disease. Nothing more is necessary in any case than to close up all the channels through which infected matter can be conveyed; but in 302 INFECTIVE DISEASES. order that this may be done close supervision by conscientious and responsible officers is required ; without it the case is hopeless. The Foot-and-mouth Disease Order of 1895 enforces notification, isolation, and disinfection, and the question of slaughter is left to the local authority. (1) A local authority may, if they think fit, cause to be slaughtered — («) Any cattle, sheep, or swine affected with foot-and-mouth disease or suspected of being so affected ; and (&) Any cattle, sheep, or swine being or having been in the same field, shed, or other place, or in the same herd or flock or otherwise in contact with animals affected with foot-and-mouth disease, or being or having been, in the opinion of the local authority, in any way exposed to the infection of foot-and-mouth disease. CHAPTER XXII. HORSE-POX.— COW-POX. CONSTITUTIONAL GREASE OR HORSE- POX. HORSE- POX is a vesicular disease of the horse communicable from animal to animal by inoculation, but never infectious. It is communicable by inoculation to man, and the attenuated virus produces phenomena indistinguishable from the results of vaccina- tion with cow-pox lymph. The existence of this disease of the horse had long been known to farmers and farriers, but Jenner was the first to draw attention to it in writing. " There is a disease to which the horse from his state of domestication is frequently subject. The farriers have termed it the grease ; it is an inflammation and swelling in the heel accompanied at its commencement with small cracks and fissures, from which issues matter possessing properties of a very peculiar kind." Jenner gave several instances in which this disease was communicated to man and to cows. Thus, a man named Merret attended to some horses with sore heels and also milked the cows. The cows were infected, and the man had several sores upon his hands. William Smith, on another farm, attended to horses with sore heels and milked the cows also. The cows were infected, and on one of Smith's hands there were several ulcerated sores. Simon Nicholls applied dressings to the sore heels of one of his master's horses and at the same time milked the cows, and the cows were infected in consequence. A mare, the property of a, dairy farmer, had sore heels, and was attended to by the men of the farm, Thomas Yirgoe, William Wherret, and William Haynes. They contracted "sores on their hands, followed by inflamed lymphatic glands in the arms and axillae, shiverings succeeded by heat, lassitude, and general pains in the limbs," and the disease was also communicated to the cows. 303 304 INFECTIVE DISEASES. But Jenner's experience of this disease was not limited to cases in which the eruption occurred in the heel. He mentions a case in which — " An extensive inflammation of the erysipelatous kind appeared without any cause upon the upper part of the thigh of a sucking- colt. The inflammation continued several weeks, and at length terminated in the formation of three or four small abscesses." Those who dressed the colt also milked the cows on the farm, and communi- cated the disease to them. Subsequently, Jenner gave a more comprehensive description of this disease. "The skin of the horse' is subject to an eruptive disease of a vesicular character, which vesicle contains a limpid fluid, showing itself more commonly in the heels. The legs first become cedematous, and then fissures are observed. The skin contiguous to these fissures, when accurately examined, is seen studded with small vesicles sur- rounded by an areola. These vesicles contain the specific fluid. It is the ill- management of the horse in the stable that occasions the malady to appear more frequently in the heel than in other parts. I have detected it connected with a sore on the neck of the horse, and on the thigh of a colt." Mr. Moore, of Chalford Hill, described a case in 1797, and re- garded the disease as virulent grease. His horse was attacked with what was supposed to be ordinary " grease." A cow was subse- quently infected, and the disease communicated to the servant, who had " eruptions on his hands, face, and many other parts of the body, the pustules appearing large, and not much unlike the small- pox, for which he had been inoculated a year and a half before, and had then a very heavy burden." In 1798, Mr. Fewster, of Thornbury, met with a case of this equine malady, and wrote a very full account to Jenner of its transmission to the human subject. " William Morris, aged thirty-two, servant to Mr. Cox of Almonsbury in this county, applied to me the 2nd of April, 1798. He told me that four days before he found a stiffness and swelling in both his hands, which were so painful it was with difficulty he continued his work ; that he had been seized with pain in his head, small of the back, and limbs, and with frequent chilly fits succeeded by fever. On examination I found him still affected with these symptoms, and there was great prostration of strength. Many parts of his hands on the inside were chapped, and on the middle joint of the thumb of the right hand there was a small phagedaenic CONSTITUTIONAL GREASE OR HORSE-POX. 305 ulcer, about the size of "a large pea, discharging an ichorous fluid. On the middle finger of the same hand there was another ulcer of a similar kind. These sores were of a circular form, and he described their first appearance as being somewhat like blisters arising from a burn. He complained of excessive pain, which extended up his arm into the axilla. On the 5th of April I again saw him, and found him still complaining of pain in both his hands, nor were his febrile symptoms at all relieved. The ulcers had now spread to the size of a seven-shilling gold coin, and another ulcer, which I had not noticed before, appeared on the first joint of the forefinger of the left hand,, equally painful with that on the right. I ordered him to bathe hi& hands in warm bran and water, apply escharotics to the ulcers, and wrapped his hands up in a soft cataplasm. The next day he wa& much relieved, and in something more than a fortnight got welL He lost his nails from the thumb and fingers that were ulcerated." Mr. Tanner, a veterinary surgeon, was the first to succeed in experimentally transmitting horse- pox to the teats of a cow by inoculating some of the liquid matter from the heel of a horse. From handling the cow's teats he became infected himself, and had two- pustules on his hand, which brought on inflammation, and made him unwell for several days. The matter from the cow and from his own hand proved efficacious in infecting both human subjects and cattle. In 1801 Dr. Loy published his experiments. A butcher had painful sores from dressing a horse suffering from * grease/ and Dr. Loy succeeded in transmitting the disease to the udder of a cow. Matter was taken from the cow and inserted into the arm of a child. Dr. Loy also inoculated a child direct from a horse suffering from * grease/ and subsequently five other children from this child. From his experiments and observations Dr. Loy was led to differentiate constitutional grease from the merely local affection commonly known as the grease, and thus he explained the failure on the part of many experimenters to transmit this disease to- the cow. " This fact induces me " — he says — " to suspect that two kinds of grease exist, differing from each other in the power of giving disease to the human or brute animal ; and there is another circum- stance which renders this supposition probable. The horses that communicated the infection to their dressers were affected with a general as well as a topical disease. The animals at the commence- ment of their disease were evidently in a feverish state, from which they were relieved as soon as the complaint appeared at their heels r 20 306 INFECTIVE DISEASES. and an eruption upon the skin. The horse, too, from which the infectious matter was procured for inoculation, had a considerable indisposition, previous to the disease at his heels, which was attended, as in the others, with an eruption over the greatest part of his body; but those that did not communicate the disease at all, had a local affection only. From this perhaps may be explained the want of success attending the experiments of the gentlemen I have mentioned." Experiments with horse-pox were also made about this time on the Continent. Sacco made some observations upon this disease at Milan. Several horses were suffering from what was called giardoni, and Sacco's servant was attacked on both arms, from dressing one of his horses troubled with this disease. Several children and cows were inoculated from the horses, but without success. In another instance, a coachman went to the hospital with the eruption on his hands, and the disease was successfully communicated to three out of nine children. In 1803 Dr. Marcet described some experiments which had been made at Salonica by M. La Font. The disease was known to the farriers in Macedonia as javart. In one case, a horse was attacked with feverish symptoms that ceased as soon as the eruption appeared. The fore legs were much swelled and several ulcers formed. M. La Font took some of the discharge from an ulcer and inoculated a cow and three children, and succeeded in transmitting the disease to two of the latter. Vaccinogenic grease was observed in Paris in 1812, and Baron cites the case of a coachman who, after dressing a horse with the " grease," had a crop of pustules on his hands, from which the disease was experimentally transmitted by inoculation to two children. A series of inoculations was started from an infant who was infected from one of the scabs taken from the pustules on the hand of the coachman. In 1813 Mr. Melon, a surgeon at Lichfield, met with vaccinogenic grease in the horse, and some of the virus was sent to Jenner, who carried on a series of arm to arm equinations for some months. And again in 1817, vaccinogenic grease broke out in a farm at Wansell. The farm-servants and the cows were infected, and Jenner employed this equine matter for a series of inoculations for eight months. In 1817 Baron described a case of a young man who had not less than fifty pustules on his hands and wrists from dressing a horse with this disease, and in the following year Baron obtained some fresh equine virus from the hands of a boy who had been infected directly CONSTITUTIONAL GREASE OR HORSE-POX. 307 from a horse. The disease assumed a pustular form, and extended over both arms. In 1818 Kahlert met with this equine disease in Bohemia, and confirmed the experiments made by Loy and Sacco. Kahlert noticed that the joint of the foot was swollen, and moisture exuded from it, and that the posterior part of the pastern was slightly red, swollen and hotter than the neighbouring parts, and a clear yellowish fluid with a peculiar odour escaped. At the slightest touch the animal showed signs of pain ; the hair was stuck together. The disease was successfully transmitted to cows and from cows to children. In 1860 the horses at Bieumes, near Toulouse, were attacked by an epizootic malady ; in less than three weeks there were more than one hundred cases. According to the veterinary surgeon, M. Sarrana, the animals suffered from slight fever, rapidly followed by local symptoms, the most marked of which were swelling of the hocks, and an eruption of small pustules on the surface of the swollen parts, which -were, at the same time, hot and painful. After three to five clays there was a discharge from the pastern which continued for eight to ten days, during which the inflammation gradually diminished. The pustules dried up, and in about a fortnight the crusts with patches of hair fell off, leaving more or less marked scars. The eruption appeared at the same time on different parts of the body, especially on the nostrils, lips, buttocks, and vulva. Sarrans believed that the mares taken to the breeding establishment at Rieumes had been infected from the ropes which had been used in tying up other affected animals, and had become thereby infected with the virus of this disease. One of the mares was taken by the owner, M. Corail, to the veterinary school to be examined by M. Lafosse. About eight days after this visit significant symptoms appeared : loss of appetite, lameness, stiffness of both pastern joints, and a hot, painful swelling of the left pastern joint. The hair was staring, and there were vesicles on the skin, from which a liquid exuded having an ammoniacal odour but less foetid than the secretion in eaux auxjambes. M. Lafosse successfully transmitted the disease to cows, and from cows to children and to a horse. In 1863 the subject of vaccinogenic grease or horse-pox again received great attention in France. A student named Amyot was engaged in dressing a horse on which an operation had been per- formed. The leg which had been operated on became the seat of a very confluent eruption of horse- pox, which was followed by such an abundant flow of serosity that at first the nature of the affection was mistaken, and it was thought to be a complication of eaux aux 308 INFECTIVE DISEASES. jambes. Amyot had a wound on the dorsal aspect of the first inter- phalangeal joint of the little finger of his right hand ; in spite of this, he continued to dress the horse entrusted to his care. The wound on his finger became accidentally inoculated with the virus, which flowed in great abundance from the horse's leg. The wound was made on August 3rd, and the next day it was swollen, and rather painful. On the 5th, Amyot suffered from malaise and great weakness; on the 6th, 7th, and 8th, vesicles appeared successively on the fingers of his left hand, and on his forehead between the two eyebrows. On the 9th, these vesicles were fully developed ; those of the fingers consisted of very large epidermic bullse on a bluish-red base. On opening them, a perfectly limpid fluid escaped in such abundance that small test-tubes might have been filled with it. The vesicle on the forehead was surrounded by a bluish- red areola, within which, the epidermis, of a leaden-grey hue, was raised, and had a slight central depression. The liquid which flowed from it when it was opened, and which continued to ooze, was also very abundant and of a deep citrine colour. The vesicles which had developed on the dorsal side of Amyot's fingers were extremely painful. The incessant shooting pains, of which they were the seat, prevented him from getting any rest for three days. On the 10th, inflammation of the lymphatics followed ; both arms were swollen and very painful, with red lines indicating the course of the lymphatic vessels. The glands of the axillae were also enlarged. The lymphatic glands behind the jaws were also swollen and pain- ful. Amyot's chief sufferings were occasioned by the intense local pain caused by the vesicles on the fingers, and by the inflammation of the lymphatic vessels and glands, which continued in this state up to the 18th of August. It was only at the end of the month that the vesicles were completely cicatrised. Bouley felt very great anxiety in the presence of the grave symptoms which accompanied the eruption. The eruption on the forehead was especially a cause of great uneasiness, because glanders manifests itself in a similar way. With virus from Amyot's vesicles the disease was transmitted to cows and to children. Further, this outbreak enabled exhaustive experiments to be made, by which it was definitely established that horse-pox is never infectious, but, like cow-pox, is transmitted solely by contact. In 1880, M. Baillet, Director of the National Veterinary School of Toulouse, was informed that a contagious malady had developed CONSTITUTIONAL GREASE OR HORSE-POX. 309 in the mares, which had been served by the stallions at the breeding establishment at Rieumes, belonging to M. Mazeres. M. Peuch was delegated to investigate this outbreak, and he visited for that purpose Berat, Rieumes, and Labastide-Clermont. At Berat three mares were examined. In one, there were scars and crusts, the remains of an eruption on the lips and in the vicinity of the vulva ; in another, there were several reddish circular ulcers in the same region ; and in a third, there were dried pustules with blackish adherent crusts at the circumference of the vulva and extending over the perinseum. On the lower part of the left flank a vesicle was discovered surmounted by a crust, and when the latter was detached a sero-sanguinolerit liquid oozed from the exposed surface. M. Peuch recognised the true nature of this disease, having several times previously had the opportunity of examining mares with a vesicular eruption round the vulva after coition, which eruption he had studied from its first appearance to complete cicatrisation, and had ascertained to be horse-pox. On proceeding to Rieumes, M. Peuch inspected eleven stallions, six horses, and five asses. In one ass there were several vesicles on the right side of the penis scattered about from the base to the glans. In another ass there was a trace of a vesicle on the penis and a characteristic vesicle on the left nostril. In an old bay mare there were the remains of an eruption on the circumference of the vulva, and in an old white mare there were not only vesicles on the vulva, but in addition vesicles on the inner side of the lower lip. M. Peuch drew special attention to these cases as likely to be confounded with aphthous stomatitis, but the existence of the same eruption on other parts of the body is an important aid in making a diagnosis of horse-pox. At Labastide-Clermont one mare was particularly noticed. This mare had been served on the 19th and 21st of April, and on the occasion of the inspection, May llth, there were the remains of an eruption around the vulva, and lymphangitis existed in the right posterior limb, which was engorged, hot, and painful in its whole extent, so that the animal walked with difficulty. The proprietor had contracted the disease in attending to his mare, and exhibited a vesicle on the thumb of the right hand, excoriated and blackened, but still recognisable. Some of the crusts collected from the cases at Berat were used for inoculating a cow. The result was successful, and the disease was transmitted by inoculation to a heifer and several students and children. 310 INFECTIVE DISEASES. M. Peuch ascertained that horse-pox caused considerable alarm from the fact that the breeders regard this eruptive affection as syphilitic, and this alarm consequently brings discredit upon the breeding establishment whence the illness has spread. He was also led to appreciate the great necessity for further study of this disease in relation to dourine or maladie du coit. In 1882 M. Peuch had the opportunity of investigating a case of horse-pox in Algeria. The disease occurred in a thoroughbred Arab. There was an eruption of vesicles, and there was also an ulcer the size of a five-franc piece in the nostril. In the mouth and on the lips there were a number of small vesicles about the size of a pea. The sublingual glands were engorged, hot, and painful on pressure. The coat, in patches, on the lateral aspect of the neck, on the shoulders, the flanks, and in the hollow of the heel, was staring, giving the appearance of small paint-brushes. On passing the hand over these, vesicles could be detected partly dry and partly secreting. The disease was transmitted to cows, and from cows to about one thousand five hundred persons. Cases similar to the one just described, in which there is more or less marked ulceration of the nostril or nasal septum, must be care- fully distinguished from glanders. And again, when the sublingual glands are affected the disease may be mistaken for strangles. NATURE AND AFFINITIES. Horse-pox and human small-pox are quite distinct diseases, and the theory that horse-pox is derived from grooms or other attendants suffering from small-pox may be dismissed without further comment. Hor^e-pox is never infectious, but is communicated solely by contact — either by grooms inoculating the virus with their hands, sponges, or brushes, or by horses coming into contact with each other, and in breeding establishments by coition. Auzias Turenne, who wrote exhaustively on this subject, maintained that horse-pox came into the same category of diseases as syphilis in man. "A un point de vue, le grease pustuleux inocule offre la plus parfaite ressemblance avec la verole inoculee, par le produit des accidents secondaires. Des deux cotes nous voyons, absence de contagion par la voie de 1'atmosphere, travail local, retentissement lymphatique et ganglionaire, fermentation universelle de Forganisme, eruption gerierale et immunite acquise centre de nouvelles atteintes. " A un autre point de vue, la ressemblance avec la variole est NATURE AND AFFINITIES. 311 frappante. Mais il s'en distingue eriorniement par 1'absence de la contagiosite atmospherique." Human small-pox belongs to a different group of diseases, and has affinities rather with small-pox of sheep and cattle plague, diseases which are not only inoculable, but highly infectious. Human small-pox is an infectious disease characterised by sudden and severe fever, followed after forty-eight hours by a generalised eruption ; horse - pox commences as a local affection, and constitutional symptoms follow. Auzias Turenne, guided by analogy, described the general- ised eruptions following "grease" or horse-pox as greasides ("comme on dit syphilides"). Horse-pox and human syphilis are absolutely distinct diseases; and there is no more ground for believing that horse-pox originates in human syphilis than there is for accepting the theory that it arises from grooms suffering from small-pox. Syphilis artificially inocu- lated on the human subject only resembles the casual or intentional inoculation of virulent horse-pox. The stages of papulation, vesicu- lation, ulceration, scabbing, and the formation of a permanent scar, occur in inoculated syphilis, and if we examine Ricord's illustrations and study the experiments of Auzias Turenne, we cannot fail to be struck with the remarkable similarity to the results obtained and depicted by Jeimer. But in order to follow the argument of Auzias Turenne we must study the natural and casual horse-pox. And if we are not familiar with what has been written on this subject, and if we restrict our knowledge to the artificially cultivated horse- pox, we shall fail to recognise the disease when we meet with it, and we shall be liable to attribute the results of the full effect of the virus to accidental contamination. Another question of very great interest is the relation of horse- pox to cow-pox. Jenner first of all propounded the theory that all cow-pox arose from horse-pox, or as he termed it " the grease," and thus cow-pox and horse- pox were manifestations of the same disease. But it was established that cow-pox also arose quite independently of horse-pox, and Jenner was led to distinguish between cow-pox, a disease peculiar to the cow, and the eruptive affection transmitted to the cow from the horse, which farmers and others, by a strange perversion of terms, called the cow-pox. Whether the eruption of cow-pox can be distinguished from the eruption of horse-pox com- municated to the cow, and whether cow-pox and horse-pox are identical, or only analogous, are questions which call for further investigation. 312 INFECTIVE DISEASES. Bacteria in Horse-pox. — Outbreaks of horse-pox have not been investigated from the bacteriological point of view, and the nature of the contagium is unknown. COW-POX. Cow-pox is a vesicular disease of the teats of cows. It is never infectious, and only attacks cows in milk, the virus being transferred from cow to cow by the hand of the milker. The disease is com- municable to milkers, and the virus artificially inoculated produces what is commonly known as vaccinia. In its clinical history and epidemiology cow-pox is totally distinct from human small-pox, and the hypothetical and entirely erroneous suggestion that the disease arises from milkers suffering from human small-pox is responsible for the belief which prevailed until recently, that cow-pox was an extinct disease in this country ; but, by the author's researches, this has been shown to be a mistake. Cow-pox is not a rare disease, and it has never been found to arise from a milker suffering from small-pox. As this is a matter of great importance in discussing the etiology of the disease, the history of outbreaks and the clinical characters of cow-pox will be given in considerable detail. According to Jenner, cow-pox had been known among farmers from time immemorial. He refers to cases occurring in 1770, 1780, 1782, 1791, 1794, 1796, and 1798. In 1799 cow-pox was raging in the dairies in London, and outbreaks were investigated by Woodville, Pearson, and Bradley. In the same year cow-pox broke out at Norton Nibley, in Gloucestershire. Pearson and Aikin referred to the prevalence of cow-pox in Wilts, Somerset, Devon, Bucks, Dorset, Norfolk, Suffolk, Leicestershire, and Staffordshire ; and Barry men- tioned its prevalence in Ireland. From this time onwards, for a long period, natural cow-pox received little or no attention in this country. Fresh stocks of lymph were raised for the purposes of vaccination, but no further attention was given to studying the disease in the cow. In 1836 Leese described an outbreak of cow-pox, and in 1838 Estlin discovered an outbreak in Gloucestershire. In 1838-39 cow-pox was met with by Mr. Fox, of Cerne Abbas, and again in 1839, in Dorsetshire, by Mr. Sweeting. Ceely frequently met with cow-pox in the Vale of Aylesbury, and particularly refers to outbreaks in 1838, 1840, 1841, and 1845. But after this, outbreaks of this disease in the cow were not recorded, though several medical practitioners met with the disease and raised fresh stocks of vaccine lymph. Thus, when COW-POX. 313 inquiries were made in 1857, it was found that Mr. Donald Dalrymple, of Norwich (on two occasions), Mr. Beresford, of Nar- borough, in Leicestershire, Mr. Gorham, of Aldeburgh, Mr. Alison, of Great Retford, Mr. Coles, of Leckhampton, Mr. Rudge, of Leominster, and one or two others, had met with outbreaks of cow-pox. In 1885 cow-pox was discovered by the author in Wiltshire. The publication of the fact led to the recognition of the disease in the same year in many parts of England, and cases were met with in man in 1888 by Mr. Forty in Gloucestershire, and by Mr. Bucknill near London in 1894. In Italy, cow-pox was found by Sacco in the plains of Lombardy in 1800, and by other practitioners in 1808-9. In 1812 it was observed at Naples by Miglietta; in 1830 in Piedmont ; and in 1832 and 1843 at Rome, by Dr. Maceroni. More recently, several out- breaks of cow-pox have been met with in this country, and the stocks of vaccine lymph renewed. In France, in 1810, cow-pox was found in the department of La Meurthe, and in 1822 at Clairvaux ; at Passy, Amiens, and Rambouillet in 1836 ; at Rouen in 1839 ; at St. Illide, at St. Seine, and at Perylhac, in 1841 ; in 1842 at Pagnac; in 1843 at Deux Jumeaux, where, during the previous thirty years, several fresh stocks of lymph had been raised and circulated. The disease occurred in a cow belonging to M. Majendie in 1844, and it was found at Wasseloune, in the department of Bas Rhin, in 1845 ; it occurred in three other departments in 1846 ; at Rheims, and in the department of Eure et Loire, in 1852 ; in the arrondissement of Sancerre, and at Beziers in 1854; and at Guyonville in 1863. It broke out on farms in three villages near Nogent in 1864 (the disease was introduced by newly purchased cows ; milkers were infected, and from one of these milkers a lymph stock was established); it also occurred in 1864, at Petit Quevilly, near Rouen; and in April 1866 at Beaugency ; in 1881 at Eysines, near Bordeaux, and again at the same place in 1883 ; and in 1844 at Cerons. In Germany, as soon as attention had been drawn to the disease, cow-pox was frequently discovered. There were as many as thirty- eight outbreaks reported in one year in Wurtemberg. It is hardly necessary, after reciting these instances, to insist that cow-pox is far from being a rare disease, as many have sup- posed who are unacquainted with the literature of the subject and unfamiliar with the appearances of the natural disease in the cow. 314 INFECTIVE DISEASES. NATURAL AND CASUAL COW-POX. To appreciate the characters of the natural disease in the cowy we must dismiss from our minds the artificial disease vaccinia, for the ordinary results of vaccination stand in much the same relation to the natural disease cow-pox as the benign vesicle of variolation to natural small-pox. The description of cow-pox given by Jenner, in 1798, was the first published account. The disease in the cow was described as consisting of irregular pustules on the teats, of a palish blue colour, surrounded by an erysipelatous inflammation, and characterised by a tendency to degenerate into phagedsenic ulcers. The animals were indisposed and the secretion of milk lessened. In referring to an outbreak which occurred epizootically in London in February 1799, Dr. Bradley gave a coloured plate of the disease on the arm and fingers of a milker. The cow-pox, he said, in this instance, " appears to have been very mild, for no loss was experienced by the farmers from the deficiency of milk, as usually happens." These early descriptions were supplemented by an account of cow'-pox by Mr. Lawrence, author of A Philosophical and Practical Treatise on Horses, and on the Moral Duties of Man toward the Brute Creation. Lawrence's article on cow-pox not only affords evidence that this disease was known to those who had the care of cattle before Jenner's paper was published, but it shows that it had also been made the subject of practical observation and study by veteri- narians. Lawrence concluded by saying : " Whatever may be the fate of cow-pox inoculation, it has and will give further occasion to a pretty large and open discussion, which is always beneficial as having a tendency to produce discovery and promote improvement ; and when the public ardour for the present topic shall have become a little cool and satisfied, I hope it will be turned by enlightened men towards another, perhaps of nearly as great consequence — namely, the prevention of the original malady in the animals them- selves. Those who have witnessed it and only reflected upon the excessive filth and nastiness which must unavoidably mix with the milk in an infected dairy of cows, and the corrupt and unsalubrious state of their produce in consequence, will surely join me in that sentiment." Lawrence was almost a century before his time. Cow-pox was not again brought forward in this light until 1887-88, when the author reported the contamination of the milk at the Wiltshire farms, and NATURAL AND CASUAL COW-POX. 315 advocated the advisability of placing this disease under the Con- tagious Diseases (Animals) Act. The numerous pathological details wanting in the early accounts of cow-pox were supplied by the painstaking and laborious re- searches of Robert Oeely. From his classical papers in the Trans- actions of the Provincial Medical Association, we can obtain a complete picture of the natural disease in the cow. In Ceely's experience in the Yale of Aylesbury, outbreaks occurred at irregular intervals, most commonly appearing about the beginning or end of the spring, rarely during the height of summer. There were outbreaks at all periods from August to May and the beginning of June, cases being met with in autumn and the middle of winter, after a dry summer. The disease was occa- sionally epizootic, or occurring at times at several farms at no great distance from each other, but was more commonly sporadic or nearly solitary. It was to be seen sometimes at several contiguous farms ; at other times at one or two farms. Many years might elapse before it recurred at a given farm, although all the animals might have been changed in the meantime. Cow-pox had broken out twice in five years in a particular vicinity at two contiguous farms, while at an adjoining dairy, in all respects similar in local and other circumstances, it had not been known to exist for forty years. It was sometimes introduced into a dairy by recently purchased cows. Twice it had been known to be so introduced by milch heifers. It was considered that the disease was peculiar to the milch cow ; it came primarily while the animal was in milk, and it was casually propagated to others by the hands of the milkers. Sturks, dry heifers, dry cows, and milch cows milked by other hands, grazing in the same pastures, feeding in the same sheds, and at contiguous stalls, remained exempt from the disease. For many years, the "spontaneous" origin of cow-pox had not been doubted in the Yale of Aylesbury. In all the cases that Ceely had noticed he could never discover the probability of any other origin. Condition of Animal primarily affected. — There was much diffi- culty in determining at all times, with precision, whether this disease arose primarily in one or more individuals in the same dairy. Most commonly, however, it appeared to be solitary. The milkers believed they were able to point out the infecting individual. In two instances, there could be very little doubt on this point. In August 1838, three cows were affected with the disease. The first was attacked two months after calving and seven weeks after 316 INFECTIVE DISEASES. weaning. This animal was considered in good health, but it looked out of condition. Heat and tenderness of the teats and udder were the first noticed signs. The other two were affected in about ten days. In December 1838, in a large dairy, a milch cow slipped her calf, had heat and induration of the udder and teats, with cow-pox eruption, and subsequently leucorrhcea and greatly impaired health ; the whole dairy, consisting of forty cows, became subsequently affected, and also some of the milkers. In another dairy, at the .same time, it first appeared in a heifer soon after weaning, and in about ten or twelve days extended to five other heifers and one cow, milked in the same shed, affecting the milkers. And in another dairy thirty cows were severely affected, and also one of the milkers. It appeared to originate in a cow two months after calving. The only symptoms noticed were that the udder and teats were tumid, tender, and hot just before the disease appeared. Condition of Animals casually affected. — In some animals, it was less severe than in others, depending on the state and condition of the skin of the parts affected, and the constitution and habit of the animal. It was sometimes observed to dimmish the secretion of milk, and in most cases it commonly did actually affect the amount artificially obtained ; with this exception, and the temporary trouble, and accidents to the milk and the milkers, little else was observed ; the animal continued to feed and graze apparently as well as before. The topical effects varied very much in different individuals ; the mildness or severity being greatly influenced by temperament and condition of the animal, and especially by the state of the teats and udder, and the texture and vascularity of the skin of the parts affected. Where the udder was short, compact, and hairy, and the skin of the teats thick, smooth, tense, and entire, or scarcely at all chapped, cracked, or fissured, the animal often escaped with a mild affection, sometimes with only a single vesicle. But where the udder was voluminous, flabby, pendulous, and naked, the teats long and loose, and the skin corrugated, thin, fissured, rough, and unequal, then the animal scarcely ever escaped a copious eruption. Hence, in general, heifers suffered least, and cows most, from the milkers' manipulations. Progress of the Disease. — Cow-pox once arising or introduced, and the necessary precautions not being adopted in time, appeared in ten or twelve days on many more cows in succession, so that among twenty-five cows perhaps by the third week nearly all would be affected ; but five or six weeks or more were required before the teats were perfectly free from the disease. NATURAL AND CASUAL COW-POX. 317 Propagation by the Hand of the Milker. — Ceely was able to confirm the way in which the disease was said to spread. In December 1838, on a large dairy farm, w^here there were three milking-shedsr cow-pox broke out in the home or lower shed. The cows in this shed being troublesome, the milker from the upper shed, after milking his own cows, came to assist in this for several days, morning and evening, when in about a week some of his own cows began to exhibit the disease. It appears that, having chapped hands, he neglected washing them for three or four days at a time, and thus conveyed the disease from one shed to another. During the progress of the disease through this shed, one of the aft'ected cows, which had been attacked by the others, was removed to the middle shed, where all the animals were perfectly well. This cow, being in an advanced stage of the disease, and of course difficult to milk and dangerous to the milk- pail, was milked first in order by the juvenile milker for three or four days only, when, becoming unmanageable by him, its former milker was called in to attend exclusively to it. In less than a week, all the animals of this shed showed symptoms of the disease, though in a much milder degree than it had appeared in the other sheds, fewer manipulations having been performed by an infected hand. Topical Symptoms of the Natural Disease. — For these, Ceely was- almost always, in the early stage, compelled to depend on the obser- vations and statements of the milkers. They stated that for three or four days, without any apparent indisposition, they noticed heat and tenderness of the teats and udder, followed by irregularity and pimply hardness of these parts, especially about the bases of the teats and adjoining the vicinity of the udder ; these pimples on skins not very dark are of a red colour, and generally as large as a vetch or a pea, and quite hard, though in three or four days many of these increase to the size of a horse-bean. Milking is genera lly very painful to the animal ; the tumours rapidly increase in size, vesicate, and are soon broken by the hands of the milker. Milking now becomes a troublesome and occasionally a dangerous process Ceely adds: "It is very seldom that any person competent to judge of the nature of the ailment has access to the animal before the appearance of the disease on others of the herd, when the cow first affected presents on the teats acuminated, ovoid, or globular vesications, some entire, others broken, not infrequently two or three interfluent ; those broken have evidently a central depression with marginal induration ; those entire, being punctured, diffuse a more or less viscid amber-coloured fluid, collapse, and at once indicate the 318 INFECTIVE DISEASES. same kind of central and marginal character. They appear of various sizes, from that of a pin's head, evidently of a later date, either acuminated or depressed, to that of an almond or a filbert, or ever larger. Dark brown, or black, solid, uniform crusts, especially on the udder near the base of the teats, are visible ; at the same time, some much larger are observed on the teats ; these, however, are less regular in form and less perfect. Some are nearly detached, others quite removed, exhibiting a raw surface with a slight central slough. On the teats, the crusts are circular, oval, oblong, or irregular ; some flat, others elevated, some thin and more trans- lucent, being obviously secondary. The appearance of the disease in different stages, or at least the formation of a few vesicles at different periods, seems very evident. The swollen, raw, and en- crusted teats seem to produce uneasiness to the animal only while subjected to the tractions of the milkers, which it would appear are often nearly as effectual as usual." Referring again to the character of the vesicle, Ceely says, that " those fortunate enough to have an opportunity of watching the disease in its progress may observe that, when closely examined, they present the following characters : In animals of dark skin, at this period, the finger detects the intumescent indurations often better than the eye, but when closely examined the tumours present at their margins and towards their centres a glistening metallic lustre or leaden hue ; but this is not always the case, for occasionally they exhibit a yellowish or yellowish- white appearance." In describing the crusts in detail, Ceely says that " large black solid crusts, often more than an inch or two in length, are to be seen in different parts of these organs, some firmly adherent to a raw elevated base, others partially detached from a raw, red, and bleeding surface ; many denuded, florid, red, ulcerated surfaces, with small central sloughs secreting pus and exuding blood, the teats exceedingly tender, hot, and swollen. ... In some animals, under some circumstances, this state continues little altered till the third or fourth week, rendering the process of milking painful to the animal, and difficult arid dangerous to the milker." " In many, however, little uneasiness seems to exist. The parts gradually heal ; the crusts, although often partially or entirely renewed, ultimately separate, leaving apparently but few deep irregular cicatrices, some communicating with the tubuli lactiferi, the greater part being regular, smoothly depressed, circular, or oval." Ceely illustrated his classical memoir with a series of valuable coloured drawings. One plate is a faithful picture of the disease on NATURAL AND CASUAL COW-POX. 319 the teats as it is ordinarily met with ; the other is a composite picture, consisting of the disease as ordinarily observed in the cow, to which is superadded a number of depressed vesicles as they occur in inocu- lated cow-pox. It is, however, an improvement on a plate published by Sacco. The latter is an elaborate drawing, representing the udder and teats of a cow, with an eruption purporting to be the natural cow-pox. Jenner had described a bluish tint in the vesicles in natural cow-pox, arid Sacco deliberately represents the natural disease by a highly coloured diagrammatic illustration in which he depicts clusters of vesicles of inoculated cow-pox, coloured blue, and with a silvery lustre. Hering has given a coloured plate of the natural cow-pox. On the teats are a number of oval and circular bullous vesicles and crusts. More recently, Layet has pointed out the same characters in the cow-pox discovered near Bordeaux in 1883 and 1884. The classical characters of the inoculated disease were wanting, particu- larly the central depression. In Wiltshire, the author could only distinguish, on the cow's teats, globular and broken vesicles and thick prominent crusts and ulcers, appearances which had very little in common with the ordinary results of vaccination. The early accounts of the severe character of the disease will appear by no means exaggerated to those who have had an oppor- tunity of studying the effects on the hands of the milkers, or indeed to those who have made themselves familiar with the descriptions given by Jenner, in some of his cases : — " Joseph Merret had several sores on his hands, swelling and stiffness in each axilla, and much indisposition for several days. " Mrs. H. had sores upon her hands which were communicated to her nose, which became inflamed and very much swollen. " Sarah Wynne had cow-pox in such a violent degree that she was confined to her bed, and unable to do any work for ten days. " William Rod way was so affected by the severity of the disease that he was confined to his bed. " William Smith had several ulcerated sores on his hands, and the usual constitutional symptoms, and was affected equally severely a second and a third time. " Wrlliam Stinchcomb had his hand very severely affected with several corroding ulcers, and a considerable tumour in the axilla. " Sarah Nelmes had a large pustulous sore on the hand, and the usual symptoms. "A girl had an ulceration on the lip from frequently holding her finger to her mouth to cool the raging of a cow-pox sore by blowing upon it. 320 INFECTIVE DISEASES. " A young woman had cow-pox to a great extent, several sores which maturated having appeared on the hands and wrists. " A young woman had several large suppurations from cow-pox on the hands." Pearson in his investigations encountered, and was informed of t similar experiences. " Thomas Edinburgh was so lame from the eruption of cow-pox on the palm of the hand as to necessitate his being for some time in hospital. For three days he had suffered from pain in the armpits, which were swollen and sore to the touch. He described the disease as uncommonly painful, and of long continuance. " A servant at a farm informed Pearson that in Wiltshire and Gloucestershire the milkers were sometimes so ill as to lie in bed for several days. " Mr. Francis said that cow-pox was very apt to produce painful sores on the hands of milkers. " A servant of Mr. Francis said that cow-pox affected the hands and arms of the milkers with painful sores as large as a sixpence. " Mr. Dolling describes the disease as ' a swelling under the arm, chilly fits, etc., not different from the breeding of the small-pox. After the usual time of sickening, namely, two or three days, there is a large ulcer, not unlike a carbuncle, which discharges matter.' " Dr. Pulteney described the disease as causing 'a soreness and swell- ing of the axillary glands, as under inoculation for the small-pox, then chilliness and rigors and fevers, as in the small-pox. Two or three days- afterwards abscesses, not unlike carbuncles, appear generally on the hands and arms, which ulcerate and discharge much matter.' " Mr. Bird wrote a short account : ' It appears with red spots on the hands, which enlarge, become roundish, and suppurate, tumours take place in the armpit, the pulse grows quick, the head aches, pains are felt in the back and limbs, with sometimes vomiting and delirium.' " Annie Francis had pustules on her hands from milking cows. These pustules soon became scabs, which, falling off, discovered ulcerating and very painful sores, which were long in healing. Some milk from one of the diseased cows, having spurted on the cheek of her sister and on the breast of her mistress, produced on these parts of both persons pustules and sores similar to her own on her hands." In more recent times these descriptions have been confirmed. In 1836 cow-pox was discovered at Passy, near Paris. A black cow, in very poor condition, had cow-pox six weeks after calving. Bousquet had no opportunity of seeing the eruption in the early stage, but on examination he found reddish-brown crusts on the teats, which later gave place to puckered scars. The milk-woman, Fleury, who had had small-pox, nevertheless contracted the disease from the cow. She had several vesico -pustules on the right hand NATURAL AND CASUAL COW-POX. 321 and on her lips. A vesico-pustule, when opened with a lancet, discharged like an abscess. In a letter to Mr. Badcock, dated April 3rd, 1845, Ceely referred to another new stock of lymph raised from a milker's hand. He added : — " In the enclosed lymph I see nothing unusually severe, except on very thin skins ; although the milker's hand exhibits now rough ulcers, one on the hand deep enough to encase a bean." Recent discoveries of cow-pox in England. — After Ceely 's cases in 1840-41, no cases of casual cow-pox on the hands of milkers were recognised as such and recorded in this country for nearly fifty years. In the outbreak of cow-pox discovered by the author in December 1887, in Wiltshire, the disease was communicated to nearly all the milkers. The reader is referred to the account of this outbreak, which has already been given in the chapter on scarlet fever (p. 274). The author's researches were confirmed by Mr. Forty in 1888, and Mr. Bucknill in 1895. In June 1888 Mr. Forty, in practice at Wotton-under-Edge, Gloucestershire, reported to the Local Government Board, that at a farm at Alderley, an eruptive disease on the udder and teats was occurring amongst cows, and that the farmer's son, and other persons engaged as milkers, had contracted an eruption like that of the cows. The farmer's son had been under Mr. Forty's care suffering from an eruption, and circum-anal piles. Mr. Forty had watched the course of the eruption from papules to vesicles and scabbing, and concluded that the eruption could not be distinguished from vaccinia. Klein visited the farm, and found a number of cows with sores on the teats and udders. The sores were of various sizes and outline, mostly irregular, and covered with brown or black scabs. Those on the teats were larger and more irregular than those on the udder. Klein was shown several milkers who had had sores on one or more fingers ; one had had a bad arm with swollen axillary glands. The farmer had also contracted the eruption ; but in these persons only scabs were visible as the remnants of their sores. A girl of about twenty had taken the place of an incapacitated milker, and noticed a red pimple form on the dorsal surface of her right thumb. Eight days afterwards there was a slightly raised circular vesicle, with dark centre and pale periphery ; the centre of the vesicle was slightly depressed. It was just under half an inch in diameter ; there was peripheral redness, but no marked areola. The girl had three good vaccination marks. Klein experimented on calves with lymph from the vesicle and 322 INFECTIVE DISEASES. crusts from the cow's teats, with the result that from both sources an eruption was produced, which in appearance and course was like vaccinia. With lymph from one of the calves, a public vaccinator inoculated a number of infants, and fine vesicles developed, indis- tinguishable from vaccinia. In 1894 Mr. Bucknill met with a case in a milkman. He had been milking a cow affected with cow-pox, and on the ninth day after exposure to infection, and the seventh day after the eruption of the first papule, there were three pocks on the fore-arm. The pocks were elevated, circular, and umbilicated, with a dull, creamy-white ring at the circumference, and there was well-marked induration and extensive areola. There were four excellent marks of primary vaccination. The vesicles contained clear lymph, and re-inoculation of the arm failed to take. An attempt to re-vaccinate the man with current calf lymph produced only topical irritation. INOCULATED COW-POX. Natural or Virulent Lymph. — Severe symptoms are not limited to milkers casually infected from the cow. Under certain conditions, artificial inoculation of fresh virus from the cow reproduces the •disease without any mitigation. Thus, in Jenner's cases : — " James Phipps. The incisions assumed at their edges rather a darker hue than in variolous inoculation, and the efflorescence around them took on more of an erysipelatous look. They terminated in scabs and subsequent eschars. " Susan Phipps was inoculated from the cow by inserting matter into a superficial scratch on December 2nd. The child's arm now showed a disposition to scab, and remained nearly stationary for two or three days, when it began to run into an ulcerous state, and then commenced a febrile indisposition, accompanied with an increase of auxiliary tumour. The ulcer continued spreading near a week, during which the child continued ill, when it increased to a size nearly as large as a shilling. It began now to discharge pus ; granulations sprung up, and it healed." Jenner's lymph was employed by Mr. Cline with similar results. " The child sickened on the seventh day, and the fever, which was moderate, subsided on the eleventh. . . . The ulcer was not large enough to contain a pea." Precisely similar experiences have since been encountered, in the early removes of fresh stocks of virulent lymph. Bousquet in France, in his first trials with a new lymph, in 1836, made three punctures, but he had soon to abandon this practice, because the intensity INOCULATED COW-POX. 323 of the inflammation was sometimes so great that it spread over the entire arm as far as the glands of the axilla. In one case, the vesicles were enormous, and the inflammation so violent, that baths, poultices, fomentations, and antiphlogistic diet scarcely sufficed to reduce it. The crusts when they fell off left ulcerations which were very slow to undergo cicatrisation. In some cases, the vesicles which resulted hollowed out the skin so deeply that they left regular holes. In the following year Estlin, in England, started a stock of fresh vaccine virus from the cow, and found on inoculating children that the new lymph was extremely active. In 52 the disease was regular, ,, 1 severe erysipelas, ,, 4 erythematous eruptions of a violent character, ,, 2 highly inflamed ulcerated arms, ,, 1 no effect after twice vaccinating, „ 8 result unknown ; supposed to have been favourable. 68 Cultivated or Attenuated Lymph. — When cow-pox lymph has been mitigated by successive transmission through the human subject, or by cultivation on the belly of the calf, with careful selection of vesicles, it will produce effects which are as follows : About the end of the second day after insertion, or early on the third day, a slight papular elevation is noticeable. By the fifth or sixth day, it has become a distinct vesicle, of a bluish-white colour, with raised margin and central cup- like depression. By the eighth day, the vesicle is perfect. It is circular, pearl-coloured, distended with clear lymph, and the central depression is well marked. On the same day, or a little earlier, the areola begins to appear, and gradually extends to a diameter of from one to three inches, accompanied with induration and tumefaction of the subjacent connective tissue. After the tenth day, the areola begins to fade, and the vesicle at the same time begins to dry in the centre ; the lymph becomes opaque and gradually concretes, and by the fourteenth or fifteenth day, a hard mahogany-coloured scab is formed which contracts, dries, blackens, and falls off between the twentieth and twenty-fifth days. A circular, depressed, foveated, and sometimes radiated scar remains behind. By selecting characteristic vesicles on the calf or on the human subject, and by collecting the lymph at an early stage on the fifth, sixth, or seventh day, this artificial disease, commonly known MN 324 INFECTIVE DISEASES. vaccinia, can be kept up in this comparatively mild form. But under certain conditions, such as a peculiarity in the subject inocu- lated, or if lymph be taken too late, there will be, just as in variolatipn, tendency to revert to the full intensity of the natural virus. Bacteria in Vaccine Lymph. — Cohn, Sanderson, and Godlee described micrococci in vaccinal vesicles. Quist. and Ferre in 1883 investigated the same subject. Yoigt in 1885 distinguished three species of micrococcus — a diplococcus, a large coccus, and a third form. Bauer in the same year described the presence of bacilli and sphsero- cocci. Marotta in 1886 regarded a tetracoccus as the specific micro- organism, and Tenhot in 1887 distinguished a dozen micrococci, two bacilli, and two yeasts. In the same year Garre isolated a micrococcus which appeared to him to be the contagium, but inoculated on a child it neither produced local vesicles nor immunity ; while Guttmann pointed out three micro-organisms which appeared to be rather more constantly present than others. Pf eiffer much more fully investigated the bacteriology of vaccine lymph, and found Saccharomyces vaccinse, which was seldom present in human lymph but constantly found in calf lymph ; sarcinae, both in human and calf lymph, including Sarcina lutea, Sarcina tetragonus, Sarcina aurantiaca, Sarcina muscopus ; bacteria and bacilli were found only exceptionally in human lymph, but frequently in calf lymph. These included a bacterium corre- sponding with Proteus vulgaris. Three mice were inoculated subcutaneously with a drop of the liquefied gelatine, but the result was negative. The injection of a considerable quantity proved fatal to guinea-pigs and rabbits, a result which was probably due to ptomaine poisoning. There were also several bacilli which did not liquefy gelatine ; these were not investigated. Staphylococcus cereus albus was found very frequently, and Staphylococcus pyogenes aureus occasionally. Pure-cultivations of these micrococci inoculated on the skin of calves produced a rapid local irritation, followed by vesiculation, but without the classical characters of the vaccine vesicle. The inoculated part was com- pletely healed in three to five days. According to Pfeiffer they explain the so-called false vaccine. Micrococcus pyogenes albus was almost constantly present. Numerous other micrococci were found, but not constantly present ; vaccine lymph being a splendid medium for the growth of micrococci. Pfeiffer pointed out that the effects of Staphylococcus pyogenes aureus, albus, and citreus, and of Streptococcus pyogenes on rabbits had an important bearing upon the practice of vaccination, and he recom- INOCULATED COW-POX. 325 mended that calf lymph should be tested before use upon children by inoculation of the ear of a rabbit. If after two days no erysipelas occurs in the inoculated rabbit, the absence of streptococci may be considered as almost proved. Two or three rabbits should be inocu- lated at the same time. The author's researches into the bacteriology of vaccine lymph extended over some, years. They independently confirmed and extended the results obtained by Pfeiffer. Having on several occasions examined vaccine lymph and vaccine pus, and failed to find a specific bacterium, the author proceeded to make a more systematic examination of the different species of bacteria in samples of current vaccine lymph. Pure-cultivations were obtained by plate-cultivation, and inoculation of the surface of nutrient agar, obliquely solidified in test tubes. Various current stocks of lymph were used in the investigation. Among the specimens of calf lymph, No. 1 yielded a torula, Bacillus pyocyaneus and Bacillus subtilis; No. 2, a bacterium, a variety of proteus, Staphylococcus pyogenes aureus, and yellow bacte- rium ; No. 3, a bacterium, micrococcus, yellow bacterium, and torula ; No. 4, yellow micrococcus, white micrococcus, white torula, yellow sarcina, white diplococcus, Staphylococcus cereus albus, and a mould fungus; No. 5, yellow sarcina, Staphylococcus pyogenes aureus, yellow micrococcus, white bacillus, Staphylococcus pyogenes albus, large white micrococcus, yellow bacterium, and a white micrococcus. Among the specimens of human vaccine lymph, No. 1 contained a white micrococcus, proteus, and Staphylococcus pyogenes aureus ; No. 2, a micrococcus, a tetracoccus, a white liquefying micrococcus, and a yellow bacterium ; No. 3, white micrococcus, yellow micrococcus, Staphylococcns aureus and flavus, a bacterium, a white micrococcus, a bacillus resembling Bacillus subtilis, Staphylococcus pyogenes cereus and a brown tetracoccus. The author is familiar with these different species of bacteria, and not one of them is peculiar to vaccine lymph ; there was no bacterium constantly present in human and calf vaccine, and there was not one which could be regarded as the contagium. To sum up, most of them are well known saprophytic bacteria, and some were identical with bacteria commonly found in suppuration. Vaccine lymph is a most suitable cultivating medium for micro- organisms, and bacteria invariably got access to the contents of the vaccine vesicle. There is no evidence to be obtained by the present methods of research as to the bacterial nature of the contagium of vaccine lymph. Copeman obtained similar results, and thus con- firmed the author's conclusions. Klein and Copeman have also observed minute bacilli in calf- 326 INFECTIVE DISEASES. lymph and in variolous lymph. Numerous attempts to cultivate them in nutrient media and in the living animal failed entirely, and the identity of the bacilli could not be determined. Pfeiffer, Guarnieri, Monti, Buffer, and Plimmer have drawn attention to structures in lymph, which they believe to be of the nature of parasitic protozoa. These bodies have been studied, more especially in the tissues. They are four times the size of ordinary micrococci, and are found in the clear vacuole in the protoplasm of epithelial cells. Whether they are really parasites or altered anatomical elements has not been determined. No other conclusion can be drawn from all these observations, except that the nature of the contagium of cow-pox is unknown. ORIGIN OF COW-POX. Jenner's original theory was that cow-pox . was derived from " grease," but subsequently he distinguished between cow-pox, a disease peculiar to the cow, and " grease," a disease transmitted to the cow from the horse, and the mistake of confounding these two diseases was attributed to farmers and farriers. Thus he wrote : — " From the similarity of symptoms, both constitutional and local, between the cow-pox and the disease received from morbid matter generated by a horse, the common people in this neighbourhood when infected with this disease, through a strange perversion of terms, frequently called it the cow-pox." Jenner's theory of the origin of cow-pox has been discouraged ; so also has the view of its being a " spontaneous " disease in the cow, though Ceely, after many years of research in the Yale of Aylesbury, could never discover the probability of any other origin. Both opinions have given way to the theory that cow-pox is small-pox transmitted to the cow — an opinion advocated by Baron, and supported by an erroneous interpretation of Ceely's and Badcock's variolation experiments. Thus the cow-pox and grease of farmers and farriers no longer attracted attention in this country, and as natural cow-small-pox has never been discovered, cow-pox has been credited with being extinct. For a full discussion of this subject the reader is referred to the work by the author on the History and Pathology of Vaccination, but the variolation experiments alluded to will be briefly mentioned. In 1801 Gassner inoculated eleven cows with small-pox lymph, and succeeded in one in producing phenomena indistinguishable from the results of ordinary vaccination with cow-pox, and children were inoculated from the cow. ORIGIN OF COW-POX. 327 In 1828 Dr. McMichael reported that several physicians in Egypt had obtained similar results, and children were successfully " vaccinated." In 1836 Dr. Martin, in America, inoculated the cow's udder with variolous lymph, and by inoculating children produced an epidemic of small-pox with fatal cases. In 1839 Reiter of Munich, after fifty unsuccessful attempts, succeeded in producing a vesicle, and a child inoculated from the vesicle contracted small-pox. In 1839 Dr. Thiele, after a number of unsuccessful attempts to inoculate cows with variolous virus, succeeded in producing a vesicle with the physical characters of the vaccine vesicle, and from it a stock of lymph was raised from which over three thousand persons were inoculated. Thiele's method was to inoculate the udder with lymph, and to select for the purpose young cows which had recently calved and had delicate skins. In England Ceely succeeded by inoculating the vulva of a heifer. One of the punctures developed into an enormous vesicle, which was undoubtedly variolous. His assistant punctured his hand with the lancet which had been used to open the vesicle, and febrile symptoms appeared, followed by an eruption on the face, neck, trunk, and limbs, at first papular, then vesicular, and finally pustular. The lymph was used in children, and " vaccine " vesicles were produced. One child suffered from vomiting delirium, and extensive roseola, but there was no eruption in any other case. In 1840 Badcock of Brighton inoculated a cow successfully, and later succeeded in variolating thirty-seven out of two hundred cows upon which he experimented. In 1847 variolation of the cow was successfully performed at Berlin, but the virus produced variola, and one of the children inoculated died of confluent small-pox. In 1864 Chauveau inoculated seventeen animals with virulent small-pox lymph. Very small papules resulted, and the virus from the papules produced variola in a child, which was infectious to others. Klein in this country until recently was uniformly unsuccessful. Yoigt, Fischer, King, Eternod, Haccius, Hime and Simpson, have all succeeded in inoculating cows and producing variola-vaccine. The results of these experiments have been very generally misin- terpreted, and claimed by some as conclusive evidence of the identity of cow-pox and small-pox. Instead of the vesicle being regarded as the most attenuated form of variola, the experimenters are said to have succeeded in producing cow-pox. It is quite true that they produced phenomena indistinguishable 328 INFECTIVE DISEASES. from the phenomena of an ordinary vaccination, but that does not mean that they produced the disease cow-pox. The vesicle which followed the inoculation, whether papular or vesicular, was small-pox. Ceely, Badcock, Yoigt, and others, succeeded in ingrafting the cow with small-pox, and when suitable lymph and suitable subjects were employed, the virus was so attenuated that a benign vesicle resulted. Similar results were obtained by Sutton and Dimsdale, and identical results by Adams, Guillou, and Thiele, by inoculating the human subject with variolous lymph without first ingrafting the disease on the cow. Vaccination with variola-vaccine is simply a modification of the Suttonian system of small-pox inoculation, only in the first remove the cow is substituted for the human subject. All those who were inoculated with Ceely's, Badcock's, or Simpson's variola-vaccine, were not in the usual meaning of the word vaccinated ; they were not inoculated with cow-pox but they were variolated, and in such an extremely attenuated form that the persons so variolated do not convey the infection. By judicious selection it is thus possible to obtain a strain of lymph from variola which, by direct inoculation of the human subject or by first inoculating a cow, is deprived of infectious properties, and produces on the arm the physical characters of an ordinary vaccine vesicle. This has been regarded as a proof of the identity of small-pox and cow-pox, but it is not so. Variola, and cow-pox are not the only diseases caused by a virus which can be attenuated until only a vesicle is produced with the characters of an ordinary vaccine vesicle. The results which have been obtained with the virus of cattle plague and of sheep- pox and horse- pox have been given in previous chapters ; and no one would urge on this account that human small-pox, cattle plague, cow-pox, sheep-pox, and horse-pox are all manifestations of the same disease. Cow-pox has never been converted into human small-pox, and, in their clinical history and epidemiology, natural cow-pox and human small-pox are so different, that the comparative pathologist is no more pre- pared to admit their identity than he is prepared to admit the identity of cow-pox and sheep-pox, or small-pox and cattle plague. Protective Inoculation. — Whether vaccination of all heifers on a farm would protect them from cow-pox when they came into milk is not known, the duration of the immunity in calves afforded by vaccination having not been determined. Calves undoubtedly have an immunity after vaccination, lasting for some weeks. In 1896 Beclere, Chambon, and Meriard experimented upon the immunising power of the serum of vaccinated calves. They COW-POX AND SMALL-POX. 329 concluded from experiments on animals and children that the serum of a vaccinated calf from ten to fifty days after vaccination will give immunity against inoculated cow-pox. They further stated that, whereas the immunity given by vaccination in the ordinary way is not complete until the eighth day, the immunity obtained by injection of the immunising serum is immediate. The serum has also been credited with therapeutic properties and has, it is said, proved efficacious in cases of small-pox. Jenner believed that cow-pox did not protect against itself but protected against small-pox, and for a century this has been a subject of much controversy. The reader is referred to the Reports and conclusions of the Royal Vaccination Commission. Stamping-out System. — It would undoubtedly be an advan- tage if cow-pox were scheduled under the Contagious Diseases Animals Act. Cow-keepers and dairy-men, being anxious that their trade should not be interfered with, very commonly conceal the existence of the disease, and perhaps nothing is known about it, unless a milker infected from the cows seeks for medical advice. The contamination of the milk with lymph, pus, crusts, and sometimes blood, renders it unwholesome, and therefore precautions ought to be taken to prevent its occurrence. If the infected cows in a herd are the last to be milked, and the milker washes his hands after the milking, the disease will not spread. CHAPTER XXIII. DIPHTHERIA. DIPHTHERIA is a specific infectious disease, especially of children, characterised most commonly by inflammation, and infiltration with lymph cells and fibrine, of the mucous membrane of the fauces, pharynx, larynx and trachea, followed by necrosis of the mucous membrane and the formation of a greyish-white false membrane, the diphtheritic membrane. In some cases a diphtheritic membrane forms in the stomach, intestine, the urinary organs and in wounds. After the separation of the membrane an ulcer remains, which may gradually heal. In the superficial part of the diphtheritic membrane there are masses of bacteria including cocci, streptococci, and bacilli. The diphtheria bacilli are not found in the blood or in the internal organs. There is no doubt of the fact that diphtheria is a disease which can be communicated from one person to another ; but the question of its origin is still a vexed one. There is a close association with insanitary conditions and decaying animal and vegetable refuse, and dampness. Localities with damp houses, defective drainage, and a cold exposure, are favourable to the development of diphtheria; but that does not necessarily indicate that these conditions can originate it. On the other hand, assuming the disease to be due to a living contagium, these insanitary conditions would afford a suitable environment predisposing to the development, and facilitating the spread, of the disease. Scarlet fever and measles predispose to diphtheria; and defective sanitary conditions, causing sore throat, may indirectly act as a predisposing cause. A great many cases have been quoted to illustrate the possibility of the conveyance of diphtheria by milk, and the theory which best harmonises with all these observations is the existence of a specific bacillus, which may be readily transferred from the throat of the diseased to the healthy ; which finds also in milk a suitable soil for its growth, and by its agency may be trans- mitted to the consumer. Such a bacillus was discovered by Loffler, 330 DIPHTHERIA. 331 and may be easily obtained from the throat of diphtheritic patients in the fol- lowing manner : — Culture Outfit. — Steel rods like or- dinary knitting needles, about six inches in length, are beaten out or rough- ened at one end, and a pledget of wool is twisted round so as to form a swab. These swabs are placed in clean test- tubes, which are then plugged with cotton- wool. The test-tubes and swabs are steri- lised by heating in the hot air steriliser f or an hour at 1 50° C. The so-called culture o u t fi t consists of a small box con- taining a test-tube of blood serum and a swab. They can be always kept ready for use, and after use should be conveyed by hand for further examination. The danger of trans- m i 1 1 i n g virulent diphtheritic material by post is obvious. When the examina- tion of the tube has been completed, the f^'^^^^^^f^'^^^^f^ ^S^M£SjtKI":r FIG. 126. — FREE SURFACE OF DIPHTHERITIC LARYNX x 350 (HAMILTON). — A, Deposit of diphtheria bacillus on surface of false membrane ; B, false membrane ; (7, mucosa ; /, lymph-cells and false membrane surrounded by meshes of fibrine ; e, surface of mucosa deprived of its epithelium ; l,v, lymph-cells containing shed epithelium. 332 INFECTIVE DISEASES. culture outfit and its contents should be disinfected or destroyed. To inoculate the tubes the patient, if it is possible, should be turned to the light, the mouth well opened, the tongue depressed, and the swab, without touching the teeth or the tongue, should be passed straight to the tonsils or pharynx, and especially to the membranous exudate. The swab is carefully and quickly withdrawn, and at once very gently rubbed over the surface of the blood serum. The culture outfit is then sent to the laboratory with full particulars, and the tubes are placed in the incubator at 37° 0., and can be examined after twelve hours. If the throat has been disinfected IS S'SSl? .-.!» V"W.-V*y \ ^ ^ "Wi ^ x>\ ->• *>- " \t »-\ • " flr-f M'^t ^^^•fc » - » 1 ^ A r *- Wj,l >f *H" ft , ^ ^ * *&& *:-& f FIG. 127. — BACILLUS OF DIPHTHERIA ; FROM A CULTIVATION ox BLOOD SERUM, x 1000 (FRANKEL and PFEIFFER). before examination, this must be taken into account, as the failure to find bacilli would not then necessarily indicate a wrong diagnosis. In all undoubted cases of diphtheria, growths will be obtained either in the form of a pure-culture of the bacillus, or far more commonly there will alsolbe colonies of various bacteria, especially Streptococcus pyogenes. Bacillus of Diphtheria — Rods, straight or slightly curved. •3 to '8 /* in breadth, and 1*5 to 6'5 ft in length. They occur singly, in pairs^ sometimes in chains, and sometimes as short leptothrix forms. In some cultures very irregular forms are observed, the bacilli being swollen at one or both ends or thicker in the middle portion, or the bacillus may contain oval or spherical DESCRIPTION OF PLATE VIII. Bacillus diphtherias and Bacillus typhosus. FIG. 1. — Cover-glass preparation from a pure-cultivation of Bacillus diph- therife on blood serum ; obtained from the throat in a typical case of diphtheria. Stained with gentian-violet, x 1200. FIG. 2. — Cover-glass preparation from a pure-cultivation of Bacillus typhosus on nutrient-agar ; from the spleen in a case of typhoid fever Stained with gentian-violet, x 1200. Plate MIL /•' » • »ji ** *•"•• *•% V * •',•»* •*, •••» , t>*ol YcA-^&fe V» Fig 1. BACILLUS DIPHTHERIA . BACILLUS TYPHOSUS DIPHTHERIA. 333 elements. They differ greatly in size and shape, often in the same cultures, and still more in cultures obtained from different sources. Spore formation is unknown. In unstained preparations there are highly refractive elements which correspond with the deeply stained parts of the bacillus. They stain readily with the ordinary aniline dyes. At certain stages of their growth they stain irregularly, the protoplasm of the rod being broken up into irregular segments. The bacillus is non-motile, and does not liquefy gelatine ; it grows at 20° C., but much more readily at higher temperatures. Colonies in gelatine plate -cultivations are yellowish-brown, and opaque, granular, and circular, but with more or less irregular margin. In plate -cultivations on agar and on glycerine agar the same description applies. On the surface of gelatine the appearances depend greatly on the method of inoculation. The growth may occur in the form of a whitish film, but if a sub-culture has been prepared from broth the growth is often composed of a number of iso- lated white colonies (Fig. 128, a). On blood serum, after twelve hours the colonies appear in the form of little elevated greyish - white or pearl-grey dots, which coalesce, forming a film if the serum is moist. On the surface of 1 per cent, alkaline glycerine agar, the appearances are found to vary, and this medium is not so suitable for the cultivation of the bacillus. In slightly alkaline broth, with or without the addition of 1 per cent, grape-sugar, the culture is cloudy, or a fine granular deposit occurs along the sides and bottom of the tube, while the broth remains clear. On potato the growth is almost invisible, in the form of a dry, thin glaze. Irregular forms are very numerous on microscopical examination, whilst the rods are thicker than usual (Welch and Abbott). In milk the organisms grow readily. FlG. 128. — PUKK-CULTURES OF BACILLUS DIPHTHERIA ON GELATINE : a, isolated colonies ; b, filmy growth. 3.34 INFECTIVE DISEASES. Dried diphtheritic membrane and cultures dried on silk threads retain their vitality for several months. A broth-culture in forty-eight hours may be used for inoculating guinea-pigs. A few drops will cause death in from three to five days ; there is hypersemia and oedema at the seat of inoculation, the lymphatic glands are enlarged, there is fluid in the peritoneal, pleural and pericardial cavities, and the lungs are congested. The bacillus is found at the seat of inoculation, but not, as a rule, in the blood or internal organs. Inoculation of rabbits produces extensive local oedema, enlargement of the lymphatic glands, and death in from four days to three weeks. Roux and Yersin pointed out that in less acute cases there was paralysis of the hind limbs. Mice and rats have an immunity. Cultures lose their virulence with age, but the filtrate from old cultures contains more toxic substance than that from fresh cultures. The toxin has been described in a previous chapter (p. 46). Old cultures sterilised by heating for an hour to 60° C. or 70° C. will render guinea-pigs immune in two weeks. The toxic substance is believed to be destroyed by this process, while according to Frankel the immunity-giving substance which is also present in the culture is not affected. According to Behring's researches, the blood of immune animals contains diphtheria antitoxin, consequently the blood of an im- mune animal is capable of neutralising the toxic properties in a filtered culture, not only in the living animal but when added to the culture in a test-tube. These researches led to the employment of the serum of an immune animal as a therapeutic agent in the treat- ment of diphtheria in man (p. 58). Bacteriological Diagnosis. — The diphtheritic bacilli are not only found in the throat while the lesions exist, but they are found after all sign of the disease has disappeared. In some cases they persist for a few days, in others for three or four weeks, and in rarer cases they have been found several months afterwards. They have also been found in the throats of persons in health, especially of those who have been in contact with cases of diph- theria, such as healthy children in infected families and healthy nurses. The bacilli which persist in the throat after recovery may be virulent up to the time of their disappearance, or they may gradu- ally become attenuated, and entirely lose their pathogenic properties. The value of a microscopical examination as an aid in the diagnosis of diphtheria has been considerably exaggerated, and unless the DIPHTHERIA. 335 bacillus when isolated is tested by inoculation the test may prove to be entirely fallacious. Loffler and Yon Hoffman both found bacilli in healthy throats, and thus created doubt as to the importance of the Loffler bacillus. Hoffman found this bacillus in the throats of twenty-six out of forty-five individuals, some of them suffering from scarlet fever, measles or some other non-diphtheritic affections, while the rest were healthy. The bacilli from these sources showed slight differences in morphological and cultural characteristics, and Hoffman was unable to decide whether these bacilli were diphtheria bacilli, which had become harmless, or whether they were accidental epiphytes, belonging to a closely allied but different species. Roux and Yersjn confirmed these observations. In a hospital for children in Paris without any question of the existence of diphtheria they found the so-called pseudo- diphtheria bacilli in fifteen cases out of forty-five. In a school, in a seaside place entirely free from diphtheria, the same bacilli were found in twenty-six out of fifty-nine children. They were also found in children with simple sore throats, and in five out of seven cases in measles. Roux and Yersin concluded that these bacilli were not distinct from the Loffler bacillus. There were slight variations, but there was no constant difference except in their pa.thogenic properties. The appearance of the colonies, the growth in broth, and the peculiar morphological elements showed characters common to both, and there was, in fact, less difference than there is between attenuated anthrax and virulent anthrax in form and in cultures; but inoculations of the bacillus did not cause death, though in some cases in guinea-pigs there was marked cedema at the seat of inoculation. On the other hand, Loffler's bacilli possess different degrees of virulence, some cultures producing only temporary cedema, while others cause death in twenty-four hours. Yirulent diphtheria bacilli subjected to a current of air can in two weeks be deprived of their virulence partially, and in four weeks entirely. Weakened bacilli can be raised in virulence by the simultaneous injection of the streptococcus of erysipelas, but bacilli deprived of their virulence and bacilli originally non-virulent cannot be made to assume virulent properties. Escherich maintained that they could be distinguished by comparative cultures ; that the pseudo-diphtheria bacilli made broth alkaline, so that in forty-eight hours litmus was turned red by Loffler's bacilli and blue by the false bacilli. The bacilli themselves, according to Hoffman, are, as a rule, shorter, wider and more uniform in size. 336 INFECTIVE DISEASES. Parke and Beebe, with a view to clearing up this question, made cultures from three hundred and thirty healthy throats. They found bacilli of three varieties : bacilli characteristic in growth producing acid reaction in broth, but having no virulence ; bacilli not characteristic in growth producing an alkaline reaction in broth, not virulent ; and bacilli producing acid reaction in broth and virulent. The virulent characteristic diphtheria bacilli were found in eight cases, non- virulent diphtheria bacilli in twenty-four, and non-virulent false diphtheria bacilli in twenty-seven. They con- cluded that the eight cases must have been in contact with diphtheria, although the throats were healthy. With regard to the bacillus in the twenty-four cases they regarded it as the true diphtheria bacillus which had lost its virulence, and the bacillus found in the twenty-seven cases showing differences in size and manner of staining and the reaction produced in broth was properly designated pseudo- diphtheria bacillus. DIPHTHERITIC DISEASES IN ANIMALS. There are diptheritic diseases of the lower animals which are in some respects similar to, and, some observers maintain, identical with, human diphtheria. In pigeons there is a disease accompanied with the formation of false membranes associated with a bacillus described by Loffler. Bacterium of Diphtheria of Pigeons (Bacillus columbarum, Loffler). — Short rods with rounded ends, mostly in irregular masses. In plate-cultivations on nutrient gelatine they formed whitish patches on the surface, and compact, ball-like masses when embedded in the gelatine. They were also cultivated on blood serum and potatoes. Subcutaneous inoculation of a pure- cultivation produced in pigeons local inflammation and necrosis ; inoculation in the mucous mem- brane of the mouth gave the appearances of the original disease. Other animals were only locally affected, except mice, in which characteristic symptoms and death resulted. They were isolated from the diphtheritic exudations in pigeons, and in sections were found in the vessels of the lungs and liver. A similar disease is known to attack fowls, and there are also diseases with development of false membranes of the respiratory passages in horses, cats and swine. Outbreaks of these diseases have been said to occur in times of prevalence of diphtheria in man, and their intercom municability has been suggested. Dr. Turner supposes that diphtheria in man originates in diseases DIPHTHERITIC DISEASES IN ANIMALS. 337 simulating diphtheria in cats, pigs, and horses ; and Klein, who accepts this theory, maintains that cats suffer from genuine diphtheria, and that after death the lungs are found full of grey, consolidated lobular patches, and the kidneys are enlarged and white. Human diphtheritic membrane inoculated subcutaneously in cats produces a painful swelling in the groin, and fever, and a fatal termination in a week. The subcutaneous and muscular tissues at the seat of inoculation are hsemorrhagic and oedematous. The internal organs are congested, and in the kidneys the medulla is congested, while the cortex is fatty. A recent culture produces illness in twenty-four hours, a painful tumour forms at the seat of inoculation, and death ensues in from two days to a week. Pneumonia and fatty white kidney are found after death, and the tissues at the seat of inoculation are hsemor- rhagic, and in parts almost gangrenous. Klein found that diphtheritic membrane or a pure-culture in- oculated into the cornea after removal of the superficial epithelium produced ulceration, and in two cases perforation of the cornea r and purulent panophthalmitis. Bacilli were again recovered from the ulcer similar in cultural characters, but conspicuously shorter and thinner. An epidemic occurred amongst cats at the Brown Institution. Five out of fourteen died. The symptoms were, running from the eyes, sometimes a muco-purulent discharge, sneezing, coughing, and pulmonary trouble, resulting in emaciation and death in from one to three weeks. Aften death lobular pneumonia and large white kidney were found ; and in" one case a diphtheritic condition of the trachea, preparations of which showed diphtheria bacilli in crowds under the microscope. Klein regarded this disease as an epidemic of cat-diphtheria, and believed that the disease was possibly induced accidentally by the cats drinking milk, which was infected in the course of some other experiments with diphtheria. He states that on account of the very definite results obtained by inoculating diphtheritic, membrane and cultures of the bacillus, subcutaneously and on the cornea, and of the condition of the lung and kidney in cats naturally or experimentally infected, the disease must be con- sidered as equivalent to human diphtheria\A and the cat capable of communicating the disease to other cats, and also to human beings. These conclusions have not yet met with the acceptance cf veterinary authorities. The results of the experimental inoculations 22 338 INFECTIVE DISEASES. are certainly by no means conclusive. It does not follow from these experiments that the disease diphtheria naturally occurs in the -cat or that under ordinary circumstances cats may contract the disease from the human subject; but the experiments show that, like guinea-pigs and rabbits, cats are susceptible to the toxic effects of the extremely poisonous principles developed during the growth of Loffler's bacillus. MILK DIPHTHERIA. It has been shown that milk infected with diphtheria has been the cause of epidemics among the consumers ; there have also been epidemics apparently associated with the milk supply, in which it has not been possible to trace the source from which the milk was infected. A difficulty in tracing the origin in no way excludes the possibility of contamination from a human source. In the light of recent researches we should expect that it would be easy to overlook the source of the virus, if it be true that diphtheria may exist without .any symptoms indicating its presence, and be unrecognised until the throat has been examined for diphtheria bacilli. As this fact was unknown until quite recently, the absence of an acknowledged •case of diphtheria was taken as evidence that no diphtheria existed, .and consequently that the milk must have been infected by a diseased condition of the cow. Mr. Power, whose views upon milk scarlatina have already been referred to, endeavoured to trace the -origin of a milk epidemic to the very common disease of " garget," or mammary abscess. This idea may be dismissed without further •consideration ; but the theory of some disease existing in the cow capable of producing diphtheria in man was resumed by Dr. Cameron, who suggested that there might be an eruptive disease of the teats producing diphtheria, and by Mr. Power, who supported the theory in an investigation of a milk-diphtheria outbreak in 1886 .at Camber ley. Diphtheria in this case existed in the neighbourhood, but as the source of human infection could not be traced, attention was drawn to two cows in the herd which had recently calved, and •especially to one with chapped teats. Following this line of inquiry, Klein investigated the behaviour of milch cows to the diphtheria bacillus. Two cows were injected subcutaneously under the skin of the shoulder with a Pravaz' syringe filled with a sub-culture in broth. There was a rise of temperature, and on the third day a painful tumour, which enlarged to the size of a child's head. In about a fortnight the tumour began to decrease, and ultimately one cow MILK DIPHTHERIA. 339 died and the other was killed. Such results might have been anticipated as the result of injecting a large quantity of the toxic products of the bacillus, but certain other phenomena were observed to which importance was attached. On the fourth day, on one of the cows an eruption on the teat was first noticed, consisting of small vesicles passing into pustules and crusted ulcers. Examina- tion of the contents of the vesicle revealed the bacillus. With matter from the vesicles and pustules two calves were inoculated, and a similar vesiculation produced at the seat of inoculation. The milk of the cows was inoculated on nutrient gelatine, and produced a culture of Bacillus diphtherise. The question naturally arose whether this eruption had any connection with the original experimental inoculation. No other cows in the locality from which these cows were obtained had a similar eruption, and it was taken for granted that it was the result of the experimental inocu- lation. By accepting the possibility of this eruption being identical with the chaps on the teats of the Camberley cows, the theory was gradually built up that cows suffer from diphtheria, which manifests itself in the form of an eruptive disease of the teats, and that the disease is conveyed in the milk to the consumers. In the original experiment the bacilli were found to have multiplied abundantly in the tumour at the seat of inoculation. The eruption might have been, as admitted by Klein, a symptom of the work of the chemical poison, and the elimination of the bacilli by the milk is also possible ; but that there is in cows a vesicular disease of the teats which is the origin of human diphtheria is not accepted by veterinarians, and there is not sufficient evidence to justify the con- clusion that the infectivity of the milk in epidemics of milk diphtheria, has been proved to be due to a morbid condition of the cow. CHAPTER XXIV. TYPHOID FEVER. TYPHOID FEVER is a specific febrile disease peculiar to man, with characteristic pathological lesions in the intestine, mesenteric glands, and spleen. The Peyer's glands pass through three stages. They become swollen from infiltration of round cells in lymph follicles, due, it is supposed, to the presence of the typhoid fever bacillus. The enlargement of the lymph follicles is followed by coagulation necrosis until the entire patch becomes necrosed, and sloughs away, leaving an ulcer. The disintegration of the patch may extend in depth, and result in perforation and peritonitis, or the ulcer may heal, and a pigmented scar take the place of the Peyer's patch. The lymphatic glands are found more or less enlarged, and may be easily felt in the groin, axilla, and neck. In some cases there is a tendency to haemorrhage, followed by infarctions in the spleen and lungs, which may develop into pysemic abscesses In the mesenteric glands similar changes take place, but without ulceration. Pneumonia may occur as a pulmonary complication. The bacteria of pneumonia and Streptococcus pyogenes may be found in association with the bacillus of typhoid fever. It is now generally accepted that the disease is conveyed by water and food which have become contaminated with the virus contained in typhoid evacuations. This has been practically proved by the number of cases which have been shown to have been intimately connected with contamination of drinking water from wells and other sources, by sewers, cesspools and faulty drains, the sewage presumably having been infected with typhoid excreta ; but whether sewage independently of typhoid contamination can originate typhoid is still an open question. Accepting the former theory as a working hypothesis, we must assume that a typhoid fever bacillus exists in the intestinal evacuations, and that it must be able to retain its vitality under very varying conditions until it gains 310 TYPHOID FEVER. 341 access by the mouth to a fresh host, and by its development in the intestine, and by the absorption of its toxic products, produces the phenomena which we recognise as typhoid fever. It ' -> , FIG. 129. — TYPHOID FEVER. ILEUM OP ADULT, SHOWING SLOUGHY AND INFILTRATED PATCHES (HAMILTON). Typhoid fever is also disseminated by milk ; sewage- contaminated water having been added to the milk, or used for washing the milk cans and other vessels. 342 INFECTIVE DISEASES. Typhoid fever cannot be communicated to the lower animals. Numerous experiments have been made by feeding and by injecting typhoid stools, but with absolutely negative results. Murchison gave typhoid fever discharges to pigs, Klein experimented with rabbits, monkeys, and other animals. Motschutkowsky injected the blood from cases of typhoid into monkeys, rabbits, and other animals, but with negative results. FIG. 130.— TYPHOID BACILLI FROM A COLONY ON NUTRIENT GELATINE, x 1000 (FRANKEL AND PFEIFFER). Various micro-organisms have been described in typhoid, but the one to which most importance is attached is a bacillus which was first discovered by Eberth, but cultivated and fully described by Gaff ky. Gaffky cultivated it from typhoid evacuations, from typhoid ulcers, from the mesenteric glands, and from the spleen. It is found in scattered colonies in the spleen, and is rarely if ever present in the blood. Bacillus of Typhoid Fever.— Rods 1 to 3 /x in length, and -5 to -8 /A in breadth, and threads (Plate VIII., Fig. 2). Spore-formation has not been observed, but the protoplasm may be broken up, producing appearances which may be mistaken for spores. They are actively motile, and provided some with a single and others with very numerous flagella, which are from three to five times as long as the bacilli. They stain well with aqueous solutions FIG. 131.— TYPHOID BACILLI, x 950 (BAUMGARTEN). TYPHOID FEVER. 343 of aniline dyes, and grow well at the temperature of the room. In plate-cultivations minute colonies are visible in thirty-six to forty- eight hours ; they are circular or oval, with an irregular margin ; they appear granular by transmitted light, and are yellowish-brown in colour. Cultivated in the depth of gelatine a well-defined shiny film forms at the point of puncture, and a greyish-white filament, com- posed of closely packed colonies, develops in the track of the needle (Fig. 134). On the surface of gelatine a greyish-white translucent film forms, with sharply defined margin (Plate II., Fig. 2). On agar there is a whitish transparent layer. They flourish in milk. On potato at the temperature of the blood there is no culture visible, but FIG. 132.— FLAGELLA OF TYPHOID BACILLI, x 1000, STAINED BY LOFFLER'S METHOD (FRANKEL AND FFEIFFEB). ' the inoculated area appears moist and shining, and cover-glass pre- parations made from the potato will demonstrate that there is really a copious growth of the bacillus. This almost invisible growth is not peculiar to this micro-organism. Whether this bacillus is really peculiar to typhoid is much dis- puted. Bacilli very closely resembling it, if not actually identical, have been found under other conditions. These pseudo-typhoid bacilli are regarded by some bacteriologists as varieties resulting from the different environment afforded by a saprophytic existence. Numer- ous experiments have been made on animals with pure-cultures of the bacillus, but in the production of typhoid fever they have been no more successful than the experiments with typhoid stools. 344 INFECTIVE DISEASES. Frankel and Simmonds inoculated a number of rabbits in the vein of the ear, producing death, in some cases .in forty-eight hours. Seitz administered broth-cultures by Koch's method of introducing them into the stomach after the administration of opium in guinea-pigs, and death resulted in several instances. But in all these cases the results depended upon the poisonous products found in the FIG. 133.— COLONIES OF TYPHOID BACILLUS. Three days old. x 100 (FRANKEL AND PFEIFFER). cultivations, a similar result following the injection of sterilised cultures. An account of the products has already been given (p. 41). Cassedebat isolated three species of bacilli from water, which could be distinguished with great difficulty, and only after the most careful comparison. The bacillus which most closely resembles it is the Bacillus coli com- munis; in fact, Boux regards it as a non- pathogenic variety of the typhoid bacillus. Others claim to be able to distinguish it by careful comparison and the application of tests. Special importance is attached to potato cultures, the typhoid bacillus forming an invisible film, and Bacillus coli communis a well-marked yellowish growth. Terni pointed out that Bacillus typhosus retains its motility in media containing hydrochloric acid, while Bacillus coli communis and other bacilli resembling those of typhoid lost their motility. Schild maintained that Bacillus typhosus FIG. 134. — PURE- CULTURE OF TYPHOID BACILLI INOCULATED IN THE DEPTH OF NUTRIENT GELATINE (BAUMGAR- TEN). TYPHOID FEVER. 345 was destroyed by exposure to the vapour of formalin, while Bacillus coli eommunis and similar bacilli isolated from water gave subcul- tures after exposure for two hours. Typhoid bacilli do not give the reaction for indol, and there is no development of gas in cultures in the depth of nutrient agar containing 2 per cent, of grape-sugar. According to Miiller, sterilised milk is coagulated in twenty-four hours, at 37° C., by Bacillus coli eommunis, but not by the Bacillus typhosus until several weeks have elapsed ; and, further, cultures on acid potato give different results. The typhoid bacillus on micro- scopical examination shows marked polar staining, but Bacillus coli eommunis only shows a slight tendency of the protoplasm to break up. FIG. 135.— TYPHOID BACILLI IN A SECTION OF SPLEEN, x 800 (FLUGGE). Kitasato suggested the negative indol test, and recommended Salkowski's method. Broth-cultivations are treated with a solution of sodium or potassium nitrite : 1 cc. of the nitrite solution (-02 gr. to 100 cc. of water) is added to 10 cc. of a broth-culture after twenty- four hours in the incubator, and on adding a few drops of strong sulphuric acid, the typhoid cultures remain colourless, but cultures of bacilli apparently identical give the red colour. On the other hand, Losener maintains that he has cultivated from earth, water and healthy human evacuations bacilli which could not be distin- guished from typhoid bacilli by any of these tests. The detection of the typhoid bacillus in water has been 346 INFECTIVE DISEASES. described in another chapter (p. 147) ; but sufficient has been said to show that bacteriological reports in which it is stated that the typhoid fever bacillus has been found in water causing typhoid epidemics must be accepted with great reserve ; and further, no one is justified in stating that the typhoid fever bacillus is undoubtedly the cause of typhoid fever. It is not found in every case of typhoid, it is not found in the blood, but it is found in those tissues which are FIG. 136. — TYPHOID BACILLI IN A SECTION OF INTESTINE, INVADING THE SUBMUCOUS- (T.J.) AND MUSCULAR LAYERS (M.), x 950 (BAUMGARTEN). commonly the seat of secondary invasion of epiphytic bacteria, whose normal habitat is the intestinal canal. Lastly, as the disease does not exist in the lower animals, the crucial test cannot be applied. The etiology of typhoid fever is still enveloped in doubt, and the nature of the contagium has not yet been determined. CHAPTEK XXV. SWINE FEVER. PIG TYPHOID, or swine fever, is a highly contagious disease peculiar to swine, causing death in from ten to thirty days, associated with a nbrinous pneumonia, enlargement of, and haemorrhage into, the lymphatic glands, and characteristic ulcers of the mucous membrane of the stomach and intestines. The lesions may assume the form of extensive croupous or diphtheritic deposit, which may fill the intes- tinal tube. But the most characteristic appearance results when the lower part of the ileum and commencement of the colon is dotted all over with elevations of the mucous membrane, resembling leather buttons or iiux vomica seeds, and sometimes with concentric rings, so that they have been compared to slices of calumba root. Swine fever is difficult to detect in the early stage, and sometimes symptoms are absent altogether in animals suffering from the disease and quite capable of transmitting it ; or nothing may be noted except cough, and possibly enlargement of the inguinal glands. In typical cases the animals are noticed not to feed, to exhibit dulness, and to have occasional rigors. Partial paralysis may follow, producing unsteady gait or loss of power over the hind legs. Diarrhoea sets in, and the evacuations become blood stained. There is occasionally a diffused or patchy reddish or purplish rash on the skin. After death the appearances most commonly found are inflammation of the peritoneum, and redness and enlargement of the mesenteric glands and the lymphatic glands in the lungs. There is generally ulceration, especially of the colon and ileo-csecal valve, or a diphtheritic exudation, sometimes pale yellow, more commonly greyish or black, similar to the centres of necrosis within the ulcers. The spleen is enlarged and liver congested, and there are haemorrhages in the kidneys. As the lungs are so commonly affected, Klein proposed the name pneumo- enteritis ; but the pulmonary lesions are not constant. Indeed, the cases in which the intestines and 347 348 INFECTIVE DISEASES. lungs are simultaneously affected are not numerous, and sometimes the lungs may be found to be perfectly healthy in cases with PIG. 137.— ULCERATION OF THE INTESTINE IN A TYPICAL CASE OF SWINE-FEVER. long-standing lesion of the intestine. Old pigs may linger on for weeks, and ultimately recover, and in the meantime act as centres for the dissemination of the disease. DESCRIPTION OF PLATES IX. AND X. Swine Fever. PLATE IX. — Part of intestine from a typical case of swine fever, showing scattered ulcers and liberation of the ileo-csecal valve. PLATE X.— From the same case of swine fever. The lungs were extensively inflamed and partly consolidated, and the lymphatic glands were enlarged and of a deep red or reddish-purple colour. Plate IX. SWINE FEVER K. Croolulunk.ftei.1. Vincent Brcoks.Day &£o HateX SWINE FEVER. Tfi.neen.1 BrocTca.Day *Soa, SWINE FEVER. 349 FIG. 138.— BACILLUS OF SWINE-FEVER No. 1. (KLEIN.) Budd first pointed out that this disease might be compared to human typhoid, both diseases being attended by a peculiar ulceration of the intestinal follicles ; but the diseases are not to be considered in any sense identical or interchangeable. Bacteria in Swine Fever. —In 1877 Klein published a research in a Report to the Local Government Board, in which he claimed to have discovered bacilli characteristic of the disease. They were described as similar to Bacillus subtilis, or Bacillus anthracis, but smaller in size. These bacilli developed into long leptothrix filaments, and formed spores. It was further asserted that on inoculation, cultures produced lesions indicative of swine fever ; the bacilli were also pathogenic in mice and rabbits. Later this bacillus was re- nounced in favour of another. In the following year Det- mers described a bacillus, but subsequently renounced it in favour of a micrococcus. In 1882 Pasteur maintained that the virus of swine fever in France (rouget) was a dumb-bell micrococcus, which produced the same effect in pigeons as the microbe of fowl -cholera. Though rouget or swine measles is probably a different disease, the occurrence of this micro-organism is of interest in this connection. In 1883 Klein again investigated swine fever, and discovered Bacillus No. 2, and maintained that these bacilli were found in ^ »*^=s £-j-frm * ^^ the blood, in the peritoneal and ^^ .? « , bronchial exudations ; and in the ** ^ «=» % air vesicles of the lungs, in the form of leptothrix filaments ten , FIG. 139.— BACILLUS No. 2. FROM A PREPARATION OF BRONCHIAL Mucus OF A PIG. (KLEIN.) FIG. 140.— BACILLUS No. 2. FROM AN ARTIFICIAL CULTURE. (KLEIN.) or twenty times the length of single rods. Cultivations were made on solid media. The organisms in these cultures were minute rods actively motile, occurring singly or forming chains, two or three 350 INFECTIVE DISEASES. times as long as Bacterium termo ; and in preparations made from diseased organs they were found to possess a very narrow trans- parent halo, a sort of hyaline gelatinous capsule. Inoculation of cultures failed to produce the lesions found in animals naturally infected. Two pigs were inoculated, one with a sub-culture from the swollen bronchial gland of a pig that had died of pig-typhoid, and a second with a culture obtained from the spleen of a mouse that had been inoculated from another case of swine fever. After two days the inguinal glands near the seat of inoculation became swollen, and the temperature rose slightly. After three or four weeks the animals recovered. Mice on the fifth or sixth day after inoculation showed symptoms of illness, then respiration became superficial and slow. Death occurred on the sixth or seventh day. Rabbits showed a rise of temperature, and death followed between the fifth and eighth days, the temperature falling before death. At the post-mortem ex- amination there was usually peritonitis, with copious exuda- tion. The kidney, spleen, and liver were enlarged and dark in many cases, there was red hepa- tisation of the lobes of the lungs, and generally pericarditis and haemorrhage under the pericardium. In 1885 Salmon, in the annual report of the United States Bureau of Animal Industry, published the result of his investigations into American hog cholera, which is identical with English pig typhoid. A motile figure-of-eight bacterium was isolated, each part being about twice as long as broad. The bacterium grew on nutrient gelatine without liquefying it, and on potato produced a brownish growth ; broth tubes became turbid on the following day. Colonies in plate- cultivations were oval or circular, and brownish in colour. Six pigs inoculated subcutaneously were all said to have died of hog cholera, and the bacterium was again obtained from the blood of the heart and spleen. The bacteria proved fatal to mice, rabbits, guinea-pigs, and pigeons. In 1893 Welch and Clement described the hog-cholera bacillus as variable in form, and they further stated that a culture obtained FIG. 141.— BLOOD OP FRESH SPLEEN OF A MOUSE, AFTER INOCULATION WITH (KLEIN.) BACILLUS No. 2. SWINE FEVER. 351 from Klein, while not possessing the characters originally ascribed to it, could not in its form, biological characters, or pathogenic pro- perties, be distinguished from the American hog- cholera bacillus. In 1887 pig typhoid was investigated at Marseilles by Eietsch, Jobert, and Martinaud, and a bacillus found. This grew rapidly on all the nutrient media. In gelatine a growth was obtained in twenty-four hours at 18° C. ; on blood serum and agar an opaque growth developed ; and on potato the growth was yellowish. It was asserted that a young pig was killed by a culture in twenty- two days, and that the characteristic ulcerations were observed in the intestines. In 1887 pig typhoid was prevalent in Sweden, and Bang and Selander experimented with cultures from a rabbit that died after inoculation with a fragment of spleen from a diseased pig. The bacilli were motile, varying from rods to cocci, without spore-formation and pathogenic in mice, guinea-pigs, and rabbits, but not in pigeons. Pigs fed on broth -cultures were said to have succumbed to genuine pig typhoid. In the blood they were generally found in the form of short oval bacteria, but in the blood of the heart longer rods were sometimes found. Metchnikoff described a bacillus isolated by Chantemesse from an outbreak in France, as highly polymorphic. Smith identified the hog-cholera bacillus with the bacillus found by Schiitz, and this in turn has baen identified with the bacillus of haemorrhagic septicaemia. From these researches it would appear to be probable that one of the bacteria isolated by Klein, and those found by Salmon, Smith, Bang, Welch and Schiitz are identical ; and further, that they have been identified with the bacillus of hsemorrhagic septicaemia. We may sum up the characters thus : — Bacillus of Klein, Salmon, Smith and Schiitz. — Very small rods, actively motile ; spore-formation not observed ; colonies circular and brown by transmitted light. Inoculated in the depth of gelatine faintly yellowish- white colonies develop along the track of the needle ; on the surface an opalescent film ; on potato they produce a straw- coloured layer, changing to brown. There is absence of indol in cultures containing peptone; the bacilli are fatal to mice, guinea- pigs, rabbits, and pigeons. Swine die after intravenous injection, but not, as a rule, after subcutaneous injection. According to Caneva, the bacillus obtained from the Marseilles epidemic would appear to be closely allied, if not identical, with the bacillus of ferret disease (Eberth and Schimrnelbusch), and the bacillus of Texas fever (Billings). 352 INFECTIVE DISEASES. Billings appears to have isolated two bacilli, one identical with the Marseilles bacillus, and the other with the hog- cholera bacillus. Bacillus of Rietsch and Jobert. — Rods about twice as long as broad, rather shorter than the bacillus of typhoid fever, longer and thicker than the American bacillus. They exhibit end-staining. They possess flagella, and are actively motile. They grow rapidly in nutrient media. They are only feebly pathogenic. They are also said to be distinguished from the bacilli of hog-cholera by producing indol in solutions containing peptone, and by causing an acid reaction in milk. Bacillus of M'Fadyean. — M'Fadyean investigated swine fever in 1895, and found bacilli which he differentiated from hog-cholera bacilli. The method employed was to inoculate the surface of nutrient agar and of potato, with fragments torn out of the centre of a lymphatic gland, with specially constructed forceps. Inocula- tions were also made from the spleen pulp, and blood, in the usual way. They are from 1 to 2 /A in length, and '6 /* in breadth. They never grow into filaments, they do not form spores, and they are actively motile. They are readily stained by the watery solutions of the aniline dyes, and are decolorised by Gram's method ; with methylene-blue they show end-staining. The bacilli grow on gelatine without liquefaction, forming a thin white line along the needle track. On agar a thin, transparent pellicle forms, which is not easily visible at first, but gradually acquires a faint greyish tint. More characteristic appearances result in plate- cultivations of gelatine-agar, at 37° C. The colonies are distinctly visible in eighteen hours, appearing when viewed by transmitted light as bluish-white, circular specks ; each colony has a dark centre and a granular margin. In broth the bacilli produce turbidity after twenty-four hours. On potato there is no visible growth, even when the surface is inoculated with an abundance of material. On solid blood serum the growth is scanty. They grow in milk without producing coagulation. They are harmless to guinea-pigs and feebly pathogenic to rabbits. Several experiments were carried out upon swine. In the first series the most rigid precautions were taken to prevent accidental infection with swine fever. Four young pigs were inoculated, upon a farm where there was no previous history of the disease. These pigs were killed, and the post-mortem examinations were said to show indications of swine fever, principally patches of diphtheritic material in the colon, and healing ulcers. The next series of pigs was inoculated at the Royal Veterinary College. Cultures were SWINE FEVER. 353 administered with milk to two pigs. Five days afterwards one pig died ; and the mesenteric glands were congested, and the mucous membrane showed spots of necrosis. The other pig was killed, and there were ulcers in the colon. In a third experiment, eight pigs were fed with milk and broth cultures. These pigs were all killed at different dates, and most of them had ulceration of the colon ; in control experiments the intestine was normal. M'Fadyean compared his bacillus with a culture of the hog- cholera bacillus, and found that the American organism grows at a lower temperature in gelatine, and colonies appear in plates much earlier. They produce a less transparent and thicker growth, and much greater turbidity in broth and a more abundant sediment. On potato they form an abundant growth at 37° C., at first yellow, later brown, with considerable resemblance to a glanders culture. Colonies upon gelatine-agar are distinguished by their opacity and sharp outline. Agar, potato, and broth cultures of the American organism consist of short ovoid forms like the bacilli of fowl cholera, while the bacillus isolated by M'Fadyean has a closer resemblance to the bacillus of glanders. M'Fadyean asserts that the American organism is not pathogenic to the pig. Pigs after feeding on broth cultures remained healthy, and showed no trace of swine fever when killed from one to three weeks afterwards. On the other hand, broth cultures of his bacillus produced the characteristic ulceration of the bowel. M'Fadyean claims, therefore, to have discovered the true pathogenic organism of swine fever. He does not appear to have compared this bacillus with that obtained from the epidemic of swine fever at Marseilles. From the description of the mor- phological and other details there seems to be a close resemblance between the two. Not less than three and possibly four species of bacilli have been cultivated from swine fever, two at different times by Klein, one by Reitsch Jobert and Martinaud, and one by M'Fadyean ; and cultures of all these bacilli have been credited with producing swine fever in experimental animals, and each one has been pronounced to be the contagium of the disease. We must conclude either that contaminated cultures were inoculated in some cases, or, what is far more probable, the swine fever which resulted in experimental animals was due to accidental infection ; and until a bacillus has been cultivated from swine fever from which another investigator can prepare sub-cultures, and with those sub-cultures produce the typical u Iterations of swine fever in pigs on a farm, or on premises 23 354 INFECTIVE DISEASES. in which swine fever is unknown, we are justified in concluding that the contagium has not yet been discovered. The mistakes which are likely to occur when the same investi- gator isolates bacilli from cases of swine fever, and subsequently inoculates or feeds healthy swine, cannot be better illustrated than by quoting from a leaflet issued by the Board of Agriculture, pointing out the great precautions necessary to prevent accidental infection. "There seems reason to believe that the disease is not infre- quently introduced by means of persons who have been in contact with diseased animals. Pig owners, therefore, are advised to prevent strangers from at any time approaching their pigs, and should the admission to the premises of spayers or castrators be necessary, those persons should be required, before approaching the animals, to thoroughly wash their hands with soap and water, and to wash and •disinfect their boots with a solution of carbolic acid and water, or .some other suitable disinfectant. Such persons might also with advantage be required to wear, while operating, a waterproof apron, which should be washed and disinfected before the wearer is per- mitted to approach the animals to be operated on." Protective Inoculation. — The experiments of Salmon and of Schweinitz have been referred to in another chapter (pp. 41, 46). A method of protective inoculation was attempted in America, but the experiments were unsuccessful, and the plan was abandoned. Stamping -out System. — Notification is compulsory, and the order in force is the Swine Fever Order of 1896, but the stamping- out system has not been applied in a thoroughly satisfactory manner, .and the disease is still very prevalent. CHAPTER XXVI. SWINE MEASLES. — DISTEMPER IN DOGS. — EPIDEMIC DISEASE OF FERRETS. — EPIDEMIC DISEASE OF MICE. SWINE MEASLES. SWINE MEASLES, or swine erysipelas, is described as an acute, in- fectious disease of swine which is very prevalent in France and Germany, but is included in this country in the term " swine fever." According to some, it is a distinct disease, and distinguished from pig- typhoid by absence of the ulceration of the intestines which is so characteristic of that disease ; while, according to others, ulceration of the intestine and ileo-ccecal valve may be found post-mortem. The onset of the symptoms, as in pig typhoid, is very rapid ; the animals cease to feed, and show other general signs of illness ; the voice is hoarse, and there is a rapid rise of temperature. On the neck, chest, and abdomen, red patches make their appearance, which extend and coalesce, and change to a dark reddish or brownish colour. These symptoms may be followed by convulsions, and sometimes by paralysis of the hind legs; and death occurs in from one to four days. It is especially a disease of young pigs, and from 50 to 60 per cent, of infected animals die. On post-mortem examination there is haemorrhage and oedema in the patches of the skin, the lymphatic glands are swollen and dark red, the peritoneum is ecchymosed, the intestinal mucous membrane is congested and swollen, and the solitary follicles and Peyer's patches are prominent, and in the neighbourhood of the ileo- ccecal valve there are, according to Fliigge, ulcers of considerable size. The liver and spleen are congested and enlarged. Pasteur investi- gated swine measles or rouget, and described a figure-of-eight micro- coccus, which he believed to be the contagium of this disease. This organism appears to be identical with the bacterium of hsemorrhagic .septicaemia, which is also commonly found in pig-typhoid. In experimenting with the virus obtained from the spleen 365 356 INFECTIVE DISEASES. Pasteur found that, by successive inoculation of rabbits, the virulence was exalted for rabbits, but attenuated for swine, and the virus which had thus been passed through the rabbit was used as a vaccine for swine, to protect them against virulent erysipelas. Pasteur found that by passing the virus through pigeons it was made more virulent for swine. In the blood, and the juice of the internal organs, and of the lymph glands, Schiitz found a minute bacillus identical with the bacillus of mouse septicaemia. Bacillus of Swine Erysipelas (Schiitz).— Extremely minute rods '6 to 1'8 //. in length, morphologically and in cultural charac- ters identical with the bacillus of mouse septicaemia. Filaments and involution forms. Spore-formation present. House mice if inoculated with a pure culture die in two to four days. Pigeons are also very susceptible. Fowls and guinea-pigs FIG. 142.— BACILLI OF SWINE FIG. 143.— BLOOD OF PIGEON INOCULATED ERYSIPELAS (BAUMGARTEN). WITH BACILLI OF SWINE ERYSIPELAS, x 600 (SCHUTZ). are immune. Rabbits after inoculation of the ear suffer from erysipelatous inflammation, identical with that produced by inocu- lation of the bacillus of mouse septicaemia. The bacilli are also pathogenic in swine and sheep. Protective Inoculation. — With Pasteur's vaccine immunity is said to be produced which lasts about a year. Schiitz and Schottelius found the minute bacilli in Pasteur's vaccine, which they had already found in cases of swine erysipelas in Germany. The results of vaccination in France are said to be very satis- factory, but in test experiments in Germany they were not so favourable. Out of 119 vaccinated swine. 5 per cent, died as the result of the inoculation, while the average loss in the ordinary way is 2 per cent. Metchnikoff found that the blood of immunised rabbits was antitoxic, and Lorenz maintains that the serum of swine which SWINE FEVER. 357 have recovered from swine erysipelas is also antitoxic, and will produce immunity in other animals. The treatment introduced by Lorenz is to inject serum in the proportion of 1 cc. to every 10 kilogrammes of the weight of the animal's body. Two days afterwards -5 to 1 cc. of virulent culture is injected, and after twelve days the dose is doubled. Lorenz inoculated 294 pigs ; 12 were suffering from swine erysipelas, and of these 6 recovered and 6 died. In the opinion of the author this disease requires re-investigation, for if it be true that rouget or schweinrothlauf is associated with ulceration of the intestines, the recognition of it as a disease distinct from our English swine fever apparently rests upon the pres- ence of a bacillus, which cannot be distin- guished from the bacillus of mouse septicaemia. The question arises whether this bacillus is really the cause of a distinct disease, swine erysipelas, or, on the other hand, whether the bacillus is really the bacillus of mouse septicaemia which has been isolated from certain cases of swine fever. The bacillus of mouse septicaemia is widely distributed, and it may only be an accidental concomitant in rouget or schweinrothlauf. The presence of the bacterium of haemorrhagic septicaemia in both rouget and pig typhoid would not prove identity, as this micro-organism is un- doubtedly only secondary in both diseases. There is great need, therefore, for further careful investigation. Clinical and patho- logical observations must be made in this country, to determine whether there are really two diseases included under the name " swine fever." If this prove to be the case, we must ascertain the clinical and pathological differences between rouget and pig typhoid. How can rouget be distinguished from cases of swine fever in which there is a patchy rash, paralysis of hind legs, but no ulceration of the intestine ? Further, how is swine erysipelas with ulceration of the intestine and ileo-ccecal valve to be distinguished from an ordinary case of pig typhoid ? FIG. 144.— PURE-CULTURE IN NUTRIENT GELA- TINE OF BACILLI IN SWINE ERYSIPELAS (BAUMGARTEN). 358 INFECTIVE DISEASES. DISTEMPER IN DOGS. Distemper is an infectious febrile disease of dogs, characterised by bronchial catarrh and discharge from the eyes. Bronchitis and pneumonia may supervene, or there may be intestinal catarrh ter- minating in dysenteric diarrhoea, sometimes complicated by jaundice. The disease may affect the nervous system, and produce convulsive contractions of the muscles of the nose, ears, lips, and limbs. Occasionally there is an eruption, especially in animals which are out of condition. The virus exists in the discharge from the nostrils and eyes, and is given off from the lungs and the skin. One attack of the disease does not confer entire immunity ; and some dogs are completely insusceptible. Bacteria in Distemper. — Millais has isolated a micro-organism resembling the pneumococcus of Friedlander, which he believes to be the cause of the disease. The bacillus occurs with other bacteria and micrococci in the nasal discharge. Protective Inoculation. — Mixed cultures of these bacteria liquefy the gelatine, and the liquid has been used as a vaccine. When applied to the nose, it is said to produce a mild attack of distemper, which protects as much as an attack of the disease contracted naturally. These results require confirmation. Inoculation of the nasal discharge in healthy dogs has been practised, so that they may have the disease under favourable con- ditions ; but the system should not be encouraged, as dogs need not necessarily contract distemper.. Vaccination with cow-pox lymph has been advocated, but it is perfectly useless. Stamping-out System. — Dogs suffering from distemper must be completely isolated. Any straw or litter which has been in contact with a diseased dog should be burnt. Clothing, collars, chains, and the kennel or premises inhabited, must be thoroughly disinfected. The animal after recovery should be washed with carbolic soap. EPIDEMIC DISEASE OF FERRETS. Eberth and Schimmelbusch investigated an epidemic disease of ferrets (frettchen-seiiche), and isolated a bacillus, which in mor- phology and cultivation agrees very closely with the bacillus of hsemorrhagic septicaemia. EPIDEMIC DISEASE OF MICE. Lbffler investigated an epidemic disease which occurred in mice kept in confinement, and isolated a bacillus resembling Bacillus typhosus. EPIDEMIC DISEASE OF MICE. 359 Bacillus Typhi Murium. — Rods varying in length ; and fila- ments ; motile ; flagellated. The colonies are circular, brownish and granular on the surface of obliquely solidified gelatine. The bacteria inoculated on the surface produce a greyish-white semi- transparent growth, and on agar and potato the appearance of the growth is very similar. They can be cultivated readily in milk and in broth. White and field mice are killed in from one to two weeks, when given bread moistened with a culture. Lofner claims to have used this method with success in Thessaly, where there was a plague of field mice causing great losses to agriculturists. CHAPTEK XXVII. ASIATIC CHOLERA. — CHOLERA NOSTRAS. — CHOLERAIC DIARRHCEA FROM MEAT-POISONING. — DYSENTERY. — CHOLERAIC DIARRHCEA IN FOWLS. ASIATIC CHOLERA. THERE are several diseases in man associated with diarrhoea, which have certain characters in common, but are totally distinct. They include Asiatic cholera, cholera nostras, dysentery, and choleraic diarrhoea. Asiatic cholera is an endemic disease of the Delta of the Ganges, a locality which has become notorious as the home of cholera. Cholera is a filth disease ; and the accumulation of filth on the banks of the Ganges, with contamination of the water, and the climate, afford most favourable conditions for the development of the cholera virus. Four great cholera epidemics have originated in, and spread from, India : in 1817, in 1826, in 1846, and in 1865. Cholera follows the routes of pilgrims and caravans, and now, owing to the rapid means of communication by steamers and railways, it spreads to the most distant parts of the world, covering in a few weeks or days distances which in former times could only be traversed in several months or even years. In 1892 the epidemic passed from India, through Afghanistan, to Russia in Asia, and quickly spread westwards along the route of the trans-Caspian railway ; and all this occurred within the space of a few weeks. By Russian emigrants it was carried to Hamburg and Antwerp ; and the virus, finding a suitable environment in the former place, produced a severe epidemic there. Thus, in about three months, it was brought into close proximity with England. Mecca is one of the great infective centres of the world, for there all the conditions are found for the propagation of cholera, including filth, overcrowding, and the water of the famous Holy Well, which is used for ablutions and drinking purposes. The return of 860 ASIATIC CHOLERA. 361 the pilgrims to Egypt, and the proximity of England to Egypt, necessitate the greatest possible precautions to prevent the intro- duction of the disease into this country. In 1884 a German Commission was sent out to India, and Koch discovered a micro-organism which he described as a curved or comma-shaped bacillus, and pronounced to be the contagium of this disease. FIG. 145.— COVER-GLASS PREPARATION OF A DROP OF MEAT INFUSION, containing a pure-cultivation of comma-bacilli, with (a) spirilliform threads, x 600. (Kocn.) Spirillum choleras Asiatics (Comma-bacillus, Koch). — Curved rods, spirilla, and threads. The curved rods or commas are about half the length of a tubercle-bacillus. They occur isolated, or attached to each other forming S~snaPecl organisms or longer screw-forms, the latter resembling the spirilla of relapsing fever. FIG. 146.— ARTHROSPORES ; («) Comma-bacillus breaking up into spheres; (b, c), formation of spheres in spiral forms ; (d, e\ groups of spheres ; (/) spirilla with spheres from an old cultivation; (ff) germination of the spheres. (HUEPPE.) Finally they may develop into spirilliform threads. In old cultiva- tions threads are found with swellings or irregularities (Fig. 148). The commas are actively motile, and possess flagella (Fig. 147). Their movements) and development into spirilla may be studied in drop- cultivations. Arthrospore formation has been described by Hueppe (Fig. 146). In plate -cultivations, at a temperature of from 362 INFECTIVE DISEASES. 16d to 20° C., the colonies develop as little specks, which begin to be visible after about twenty-four hours. Examined with a low power, and a small diaphragm, these colonies have the following characteristics. They appear as little masses, granular, and FIG. 147. — FLAGELLA OF COMMA-BACILLI ; STAINED BY"; LOPFLER'S METHOD (FRANKEL ' AND PFEIFFER). yellowish-white in colour, and sometimes very"- faintlyi tinged « with red, which have liquefied the gelatine, and sunk down to the bottom of the resulting excavations. In test-tubes of slightly alkaline nutrient gelatine (10 per cent.), / Of, FIG. 148.— INVOLUTION FORMS, x 700 FIG. 149.— COLONIES OF COMMA-BACILLI (VAN ERMENGEM). ON NUTRIENT GELATINE, NATURAL SIZE (KOCH). the appearance of the growth is very striking. In typical cultures it begins to be visible in about twenty-four hours. Liquefaction sets in very slowly, commencing at the top of the needle track ASIATIC CHOLERA. 363 around an enclosed bubble of air, and forming a funnel continuous with the lower part of the growth ; the latter preserves for several days its resemblance to a white thread (Plate II., Fig. 1). In about eight days, however, lique- faction takes place along the whole of the needle track. On the surface of agar-agar the cultivation develops as a white, semi-transparent layer, with well-defined margin. The appearance on blood serum is very similar ; liquefaction very slowly takes place. In broth they form a wrinkled film on the surface, there is a rapid and abundant growth at the temperature of the FIG. 150. COLONIES OF KOCH'S COMMA- BACILLI, x 60. FIG. 151.— COVER-GLASS PREPARATION FROM THE CONTENTS OF A CHOLERA INTESTINE, x 600. (a) Remains of the epithelial cells; (b) Comma-bacillus; (c) Group of comma-bacilli (Koch). FIG. 152.— COVER-GLASS PREPARATION OF CHOLERA DEJECTA ON DAMP LINEN (two days old), x 600. Great prolife- ration of the bacilli with spirilla (a) (Koch). blood, and the same applies to sterilised milk ; and they will even multiply in sterilised water. In potato- cultivations the microbe will only grow at the temperature of the blood (37° C.), forming a slightly brown, transparent layer. Inoculation of a cultivation of the bacillus in the duodenum of guinea-pigs, with and without 364 INFECTIVE DISEASES. ligation of the bile-duct, has given positive results. More recently these results have been confirmed by the following method : Five cc. of a 5 per cent, solution of potash were injected into the stomach of a guinea-pig, and twenty minutes after, 10 cc. of a cultivation of comma-bacilli, diffused in broth, were similarly intro- duced. Simultaneously with the latter, an injection of tincture of opium was made into the abdominal cavity, in the proportion of 1 cc. for every 200 grammes weight of the animal. Those who have had success with inoculation experiments maintain that choleraic symptoms were produced without any trace of peritonitis or putrid infection, and that the comma -bacilli of Koch were again found in the intestinal contents, and fresh cultivations established. FIG. 153.— SECTION OF THE Mucous MEMBRANE OF A CHOLERA INTESTINE, x 600. A tubular gland (a) is divided transversely ; in its interior (6) and between the epithelium and the basement membrane (c) are numerous comma-bacilli (Koch). On the other hand, these results have been disputed, the fatal effects of the inoculation attributed to septicsemic poisoning, and the proliferation of the bacilli considered to be dependent upon an abnormal condition of the intestines, induced by the injection of tincture of opium. It has, however, been shown that these organisms, like several others which have been isolated from intestinal dis- charges, produce definite poisonous substances. The comma-bacilli were found in the superficial necrosed layer of the intestine, in the mucous flakes and liquid contents of the intestinal canal of cases of Asiatic cholera. It is stated that they were, also detected ASIATIC CHOLERA. 365 in a tank which contained the water supply of a neighbourhood where cholera cases occurred; but comma-shaped organisms are frequently present in sewage-contaminated water. Koch's comma- bacilli are aerobic, and their development is arrested by deprivation of oxygen. They are destroyed by drying on. a cover glass, but retain their vitality longer when dried on silk threads. Cultures are sterilised by exposure for fifteen minutes to 55° C., and by various antiseptic substances. FIG. 154.— PURE-CULTIVATIONS IN NUTRIENT GELATINE, a, KOCH'S CHOLERA BACILLUS, twenty-four hours old. I, TINKLER'S BACILLUS, twenty-four hours old. METHODS OF STAINING THE COMMA-BACILLI OF KOCH. In cover-glass preparations they may be well stained in the ordinary way, with an aqueous solution of methyl-violet or fuchsine, or by the rapid method, without passing through the flame (p. 85, Babes' method). Nicati and Reitsch's method. A small quantity of the stools, or of the scraping of the intestinal mucous membrane, is spread out on a glass slide and dried, then steeped during some seconds in sublimate solution, or in osmic acid (1 to 100). It is then stained by immersion in fuchsine-aniline solution (1 or 2 grammes of Bale fuchsine dissolved in a saturated aqueous solution of aniline), washed, dried, and mounted in Canada balsam. 366 INFECTIVE DISEASES. In sections of the intestine their presence may be demonstrated by : — (a) Koch's method. Sections of the intestine, which must be well hardened in absolute alcohol, are left for twenty-four hours in a strong, watery solution of methylene-blue, or for a shorter time if the solution is warmed ; then treated in the usual way. (6) Babes' method. Sections, preferably from a recent case of cholera, and made as soon as possible after death, are left for twenty-four hours in an aqueous solution of f uchsine, then washed in distilled water, faintly acidulated with acetic acid, or in sublimate solution 1 in 1000, passed rapidly through alcohol, and finally treated in the usual way. Klein investigated cholera in India, and does not accept Koch's conclusions. With regard to the inoculation experiments, Klein believes that the living choleraic comma -bacilli, even if introduced in large numbers into the small intestine, are quite innocuous, but capable of great multiplication if the intestine is previously, from some cause or another, diseased ; the chemical products of the comma -bacilli then act as poisons analogous to the ptomaines obtained from other putrefactive bacteria. The observations made by Roy, Brown, and Sherrington, in Spain, tended to confirm Koch's views. Comma-bacilli were found to be present, in some cases, in enormous numbers, and the frequency of their occurrence led these observers to believe that they must bear some relation to the disease. At the same time, as they failed to find them in all cases, they regarded the existence of a causal relation as not proven. They failed to find the Naples bacterium, or a small, straight bacillus noted by Klein ; and they drew attention to certain FIG. 155. -COMMA-SHAPED ORGANISMS peculiar mycelium-like threads WITH OTHER BACTERIA IN SEWAGE- CONTAMINATED WATER, x 1200. m the mucous membrane of the intestines ; but these cannot be considered to have any significance. Methylene-blue has been employed by Koch and others, including the author, for staining sections of the intestine from cholera cases, and had they been constantly present, it is hardly possible that such striking objects could have been overlooked. Again, we must bear in mind that hypho-mycetous fungi occasionally have been found to occur sapro- phytically in the intestinal canal, as well as in the lungs, external auditory meatus, and elsewhere. Cunningham, of Calcutta, maintains ASIATIC CHOLERA. 367 that Koch's comma-bacilli are not constantly found ; and that the comma -bacilli obtained from typical cholera cases show a great variation in cultivation, and cannot be distinguished from comma-bacilli from other sources. Cunningham asserts that comma-bacilli resembling Koch's are found in the intestine in * health. Sternberg, on the other hand, made a number of examina- tions of the evacuations of yellow , , ,,, . ,. FIG. 156.— COMMA-BACILLI OF THE MOUTH, fever patients and healthy mdi- x m (VAN ERMENGEM). viduals, and failed to find any micro-organism resembling the cholera spirillum. Various comma-bacilli have been isolated from different sources and compared with Koch's comma -bacillus. Comma-bacilli have been found in the mouth by Lewis ; in cholera nostras by Finkler and J^yf Prior ; in cheese by Deneke ; »'l)/(/r' V. ^ i11 nay infusion and sewage by Weibel ; in the intestines of fowls by Gamaleia, and in water by Sanarelli. Whether the comma-bacillus FIG. 157.— FINKLER'S COMMA-BACILLI ; is the cause of cholera or FROM CHOLERA NOSTRAS, x 700 not? its detection is an aid in (FLUGGE). ,. . T,. , ,. diagnosis. It we are dealing with a case alleged to be one of Asiatic cholera, and a micro- organism is found in the intestinal evacuations, which can be differentiated from the comma-bacillus described by Finkler in cholera nostras, and identified with the comma-bacillus de- -^ . scribed by Koch, we are justified ^> ^ \ in regarding the case as one of "•» * p"* ^v'Z.i' •* "* Asiatic cholera. But we cannot ^~^* • ^ ^~* diagnose Koch's comma -bacillus, '^^NfV* f ^X with certainty, unless we know the source of the culture. The FJG m_DENEKE>s COMMA-BACILLI, clinical symptoms of cholera in FROM CHEESE, x 700 (FLUGGE). man, and especially the presence of rice-water stools, must be taken into account, together with the biological, morphological, and chemical characteristics of the bacilli which are found to be present. There are several 368 INFECTIVE DISEASES. chemical tests which can be applied to cultures. According to Frankel, the Bujwid- Dunham test can be relied upon to distin- guish Koch's comma-bacillus from the comma-bacillus of Finkler- Prior (cholera nostras), and from those found by Gamaleia. The comma-bacilli are inoculated in broth containing peptone, and, after twelve hours in the incubator, a drop of strong sulphuric acid added to the culture will produce a red colour, owing to the presence of indol. A test which distinguishes Koch's comma- bacillus from Finkler- Prior's and Deneke's was introduced by Cahen. A solution of litmus is added to the broth, and the culture placed in the incubator, until the following day ; in the case of Koch's commas, the colour will have disappeared. Koch points out that in the bacteriological diagnosis of cholera the first step is to examine the mucus in the evacuations, or in the intestine if the examination is made after death. Cover-glass preparations should be stained with dilute Ziehl-Neelsen solution. Cultures are next made in peptone, and in eight hours will give the indol reaction. In twenty-four hours the colonies may be examined on plate-cultivations. The peptone cultures are prepared by adding a trace of the choleraic evacuations, or of mucus containing the bacilli, to a sterilised 1 per cent, solution of peptone, with '5 to 1 per cent, of common salt. The solution must be alkaline, and the culture is placed in the incubator at 37° C. The pathogenic effects can be ascertained by diffusing the bacilli from an agar-culture in broth, and injecting it into the peritoneal cavity. Toxic Products. — Brieger isolated several toxic products which he had found in association with putrefaction, such as cadaverin and putrescin ; but there were also present two new toxic substances, one producing cramps and muscular tremors in inoculated animals, and the other lowering the temperature and depressing the action of the heart. Later, Brieger in conjunction with Frankel, succeeded in isolating a tox-albumin from pure cultures. Guinea-pigs were killed in two or three days, but rabbits had an immunity. Pfeiffer found that cultures contained a poisonous principle which proved fatal to guinea-pigs in extremely minute doses. It is broken up by alcohol and by boiling, and secondary products formed, of very much mitigated virulence. Similar toxic products were obtained from cultures of both Finkler- Prior's and Metchnikoff's commas. Protective Inoculation — Haffkiiie has introduced a system of protective inoculation, which is founded on the principle of inducing the formation of antitoxins, or defensive proteids. Comma-bacilli when first cultivated from a cholera patient are not sufficiently ASIATIC CHOLERA. 369 virulent, and the virulence is increased by cultivation in the peritoneal cavities of a succession of guinea-pigs. This successive cultivation is carried on until a virus is obtained which proves fatal in a few hours when inoculated into the peritoneum. A culture from the peritoneum is obtained on an agar plate- cultivation, and a pure sub-culture on agar is thoroughly shaken up with broth. This constitutes the vaccinating fluid. It may be used as a living vaccine, or the comma-bacilli killed by the addition of carbolic acid. Haffkine, having studied the pathological and physiological effects on some sixty persons, mostly scientists interested in the subject, and finding the treatment to be harmless, transferred his operations to localities in India affected by cholera. The inhabitants of the northern part of India were the first to come forward and submit themselves to the inoculation. In the course of the first year 22,703 were inoculated in the North- West Provinces and Oudh, and in the Punjab. All classes ofi the population were included. In the second year operations were carried out in those parts of the country where cholera always prevails, and where, therefore, the method could be more satisfactorily tested. From March 1894, to July 1895, 19,473 individuals were inoculated in some of the most affected localities. From observations made at Calcutta by Dr. Simpson, from March 1894 to August 1895, cholera occurred in 36 houses containing inoculated people. There were 521 inhabitants in the infected houses, of whom 181 were inoculated from 1 to 459 days before the occurrence, while 340 remained uninoculated. The uninoculated had 45 cases with 39 deaths from cholera ; the inoculated had 4 deaths, 1 occurring 451 days after the first inoculation, and 3 others from 1 to 4 days after the first inoculation. These four cases had not been re- inoculated. If the occurrences in inoculated and non-inocu- lated during the first 10 days were set aside, and those considered that occurred after the 10 days expired, then, according to Dr. Simpson, the proportion of cases was 19 '2 7 and that of deaths 17 '24 times smaller in the inoculated then in the uninoculated. Cholera broke out in the Gya gaol, and inoculations were made after 6 cases, with 5 deaths, had occurred. During the stay of the prisoners in the gaol, there were 209 uninoculated, with 7 cases and 5 deaths, and 211 inoculated, with 5 cases and 4 deaths. In July and August in the same year cholera attacked the East Lancashire Hegiment. Out of 773 men there were 133 inoculated and 640 uninoculated. 24 370 INFECTIVE DISEASES. The occurrences of cases and deaths were : — In 640 uninoculated 120 cases (18'75 %), 79 deaths (12-34 %). In 133 inoculated 18 cases (13-53 %), 13 deaths (9'77 %). These results were, it is said, due to the weakness of the vaccines procurable at that period of the work, and to the small doses used. There were a great many records kept of the results of inocula- tion of coolies on tea estates in different localities. After a summary of the results Haffkine concludes, in his Report to the Government of India, that, in his opinion, the experimental stage was not yet in so advanced a condition as to be completely closed; but that the observations made and records collected justified steps being taken to give the inoculations a more extended trial. CHOLERA NOSTRAS. Cholera nostras, English cholera, or English dysentery, produces an inflamma- tion of the mucous membrane of the bowels with croupous exudation. The large intestine is commonly affected, and the mucous membrane may be covered with small superficial ulcers. The disease is associated with severe diarrhoea. Finkler and Prior obtained a comma- bacillus from the evacuations, which they believed to be identical with the comma- bacillus found by Koch in Asiatic cholera. Koch pointed out that there were marked differences in the biological character of the two micro-organisms. Spirillum Finkler-Prior (Comma- bacillus in Cholera nostras). — Curved rods, thicker than the comma-bacillus of Koch, and spirilla. The colonies on plate- cultivations are very much larger than those of the comma -bacillus of Koch of the same age. They have a very faint yellowish-brown tinge, a well-defined border, and a distinctly granular appearance. They liquefy nutrient gelatine very rapidly, so that the first plate of a series is, as a rule, completely liquefied on the day following inoculation, and the second plate in two or three days more. In a test-tube cultivation in nutrient gelatine the appearances are especially characteristic : the gelatine is very rapidly FIG. 159.— PURE-CULTIVATION OF THE SPIRILLUM ^INKLER- PRIOR, IN NUTRIENT GELA- TINE. In thirty-six hours. CHOLERAIC DIARRHCEA FROM MEAT POISONING. 371 liquefied along the whole track of the needle, so that the cultivation resembles a conical sack, or the finger of a glove turned inside out. On a sloping surface of nutrient agar-agar a white moist layer forms very quickly. On potato they grow at the ordinary temperature of the air, producing a brownish layer and corrosion of the surface of the potato. They have been shown to be pathogenic. CHOLERAIC DIARRHCEA FROM MEAT POISONING. There are two varieties of choleraic diarrhoea from meat poisoning, and both are associated with vomiting, diarrhoea, pain in the abdo- men, in severe cases followed by suppression of urine, collapse, and death. These conditions are brought about by poisonous foods, and include those cases of poisoning by tinned meats, pork pies, hams, cheese, sardines, and other articles of food improperly prepared. In most cases putrefaction has taken place, owing to the action of various bacteria. Associated with their growth we find highly poisonous substances, but no bacteria are found in the body in these cases. They are all due to chemical poisoning ; but Klein has also described cases of poisoning due to the growth of bacteria without the presence of putrefaction. The latter were of the nature of an infectious disease. In the Welbeck poisoning cases, described by Ballard, the poisonous hams contained a short bacillus, which was also found in the kidney and spleen in the fatal cases in man. In the Carlisle epidemic, which was due to poisonous pork pies, the pork and gravy stock proved fatal to mice, and from the infected mice a bacillus was cultivated, which, administered to mice by feeding or subcutaneous inoculation, produced enteritis, diarrhoea, and congestion of the lungs. Gartner cultivated Bacillus enteritidis from the spleen in a fatal case of meat poisoning. Gaffky obtained a similar bacillus in cases of gastro-enteritis, following the consumption of meat and sausages, which had been made of horseflesh. Bacillus of Choleraic Diarrhoea from Meat-poisoning (Klein). — Rods from 3 to 9 /A in langth, 1*3 /x wide, rounded at their extremities, singly or in chains of two. Spore- formation occurs, the spores being 1 //, thick, ovtil, and situated in the centre or at the end of the rod. Feeding mice with the bacilli and inoculation produced positive results. At the autopsy, pneumonia, peritonitis, pleuritis, enlargement of the liver and spleen, and haemorrhages were observed, and bacilli were present in the blood and exudations of these animals. They 372 INFECTIVE DISEASES. occurred in the blood and juices, and especially in the glomeruli of the kidneys, of several fatal cases of choleraic diarrhoea. Bacillus enteritidis (Gartner). — Short rods in pairs, and short chains. They are motile ; spore-formation not observed. Colonies are granular, and old colonies at the margin have an appearance of FIG. 160.— TROPICAL DYSENTERY. Mucous membrane of large intestine some months after an acute attack : a,a, representing remains of mucosa ; 6,6, inter- vening parts corresponding to the muscularis (HAMILTON). powdered glass. On the surface of gelatine a thick greyish-white film develops, which in time becomes wrinkled. In the depth of gelatine a white filament forms. The gelatine is riot liquefied. On agar the film is slightly yellowish. On potato it is similar in colour, moist and shining. On blood serum it is very similar. Mice fed CHOLERAIC DIARRHCEA IN FOWLS. 373 with the bacilli die in one or two days. Subcutaneous injection is fatal in guinea-pigs and rabbits in from two to five days. Dogs, cats, and fowls are immune. The bacilli were obtained from a cow suffering from a disease associated with diarrhoea, and from the spleen of a man who died twelve hours after partaking of the flesh of this animal. DYSENTERY. Dysentery is a disease of tropical climates associated with in- flammation and ulceration of the large intestine (Fig. 160). At first the discharge from the bowel is a whitish or brownish mucus, which soon becomes blood-stained; later the evacuations become thin and watery, with altered blood clots, fragments of mucous membrane, and pieces of false membrane ; and in some cases they become purulent. The virus is believed to be in the intestinal discharges, which by contaminating water or soil may give rise to other cases. Micrococci have been found in dysentery, but the micro-organism which has received most attention is a protozoon, the Amceba coli, which will be described in another chapter. CHOLERAIC DIARRHOEA IN FOWLS. Choleraic diarrhoea in fowls, or gastro-enteritis cholerica, is an infectious disease of fowls, occurring in Russia during the summer. The disease is very like fowl-cholera. The birds are sleepy, and suffer from diarrhoea, but the temperature is not raised, as in fowl-cholera. After death there is usually an abundance of greyish liquid in the small intestine, which is stained with blood. It was investigated by Gamaleia, who found a comma-bacillus, to which he gives the name Vibrio Metchnikovi. Spirillum of Fowl- enteritis (Vibrio Metchnikovi). — Curved rods and spirilla; thicker, shorter, and more curved than Koch's commas. They are motile, and possess a single flagellum at one end. They stain with the usual dyes. Spore-formation doubtful. In plate- cultivations minute white colonies appear in from twelve to sixteen hours, and the gelatine is liquefied. The colonies in about three days resemble those of both Finkler- Prior's and Koch's comma-bacilli, some colonies being more like the one kind, and some like the other. In the depth of gelatine the growth is very much like that of Koch's comma-bacillus, possessing the characteristic air-bubble appearance. 374 INFECTIVE DISEASES. On agar a slightly yellowish growth is obtained, resembling that of Koch's commas ; on potato a yellowish -brown or chocolate layer develops after incubation at the temperature of the blood, very similar to cultures from Asiatic cholera. Broth becomes turbid, and a wrinkled film forms on the surface ; the addition of sulphuric acid gives the indol test. The spirilla grow in milk, and coagulate it ; the milk becoming strongly acid, and the casein being precipitated. They are pathogenic in chickens, pigeons, and guinea-pigs. Pigeons die in about twelve hours after a subcutaneous injection ; and the spirilla are found abundantly in blood from the heart. Guinea-pigs die from acute septicaemia in about twenty-four hours. The spirilla are found in the blood and internal organs. Inoculation of pigeons and guinea-pigs with sterilised cultures will produce immunity. CHAPTER XXVIII. TUBERCULOSIS. TUBERCULOSIS is a communicable disease of man and animals, charac- terised by the formation of new growths associated with the presence of the tubercle bacillus. Yon Bayle, in 1810, was the first to describe little growths like millet seeds, which were considered to be character- istic of consumption or phthisis. Laennec, in 1834, attached much more importance to the existence of caseous matter and classified miliary tubercle, crude tubercle, granular tubercle, and encysted tubercle, as varieties of tuberculosis. Virchow would not accept all these varieties as tubercular, and only regarded those conditions associated with the presence of miliary tubercles as genuinely tubercular. Laennec's so-called crude tubercle, for example, was simply due to pneumonic caseation. Yillemin threw entirely fresh light upon this controversy by proving that tuberculosis was a communicable disease. Rabbits and guinea-pigs, inoculated with tubercular sputum or caseous tubercle, developed miliary tubercle in a few weeks. Sanderson confirmed these experiments, and pointed out that foreign bodies would produce experimental tuberculosis in rabbits. Cohnheim also confirmed the experiments of Villemin, and maintained that tuberculosis was a specific inoculable disease, and, therefore, everything was tubercular which, on inoculation, produced tuberculosis. Koch, in 1882, announced the discovery of the tubercle bacillus, and expressed the opinion that without the tubercle bacillus there could be no tuberculosis. Tubercle was defined as tissue containing the tubercle bacillus, whatever might be the clinical manifestations of the case, or the microscopical and naked-eye appearances of the diseased parts. A tubercle is a small growth about the size of a millet seed. In the early stage it is circular, hard, grey in colour, and lustrous ; but when it undergoes necrosis and caseation it becomes soft and yellowish. In the very early stage it consists of a little collection of round 375 376 INFECTIVE DISEASES. cells, in which it is possible, though often with extreme difficulty, to demonstrate the tubercle bacillus. The cells originate in the proliferation of endothelial connective tissue and white blood cells. Later on, large oval or circular multi- nucleated cells, or giant cells, make their appearance. The tubercle bacilli are only occasionally found in the interior of human giant cells, whereas in the lower animals, in equine and bovine tuberculosis more especially, the bacilli are often present in great numbers, and very commonly in the form of conspicuous rings, visible under a low power of the microscope. FIG. 161.— TUBERCLE OF THE LUNG IN A VERY EARLY STAGE, x 400 : a, An alveolar wall ; b, blood-corpuscles in capillaries of the same ; c, blood-corpuscles extra vasated into the alveolar cavities; d, alveolar capillaries filled with blood-corpuscles carried forward by the tubercle which is growing into the alveolar cavity ; e, large endothelium-like cells, of which the tubercle in this stage is mainly composed ; /, portion of a branch of the pulmonary artery injected (HAMILTON). Whether the absence of blood-vessels or the action of the bacillus is the main factor in producing caseation, is an open question. When suppuration follows caseation, as commonly happens in tuberculosis of the lungs in man, and in experimental tuberculosis in animals, an abscess forms. In cattle there is a remarkable tendency to the formation of calcareous deposit in the caseous masses. The tubercle may not degenerate and die, but live and develop. TUBERCULOSIS. 377 The giant cells, which are more or less central, have been described as sending off processes, which, by dividing and subdividing, and FIG. 162. — PRIMARY TUBERCLE OF LUNG TWO TO THREE WEEKS OLD, x 50: a, Portion of wall of a branch of the pulmonary artery ; b,b, giant cells with concentric arrangement of fibrous tissue ; c, centre of tubercle beginning to caseate ; d, small branch of pulmonary artery seen on transverse section ; e, injected capillaries of the alveolar walls (HAMILTON). interlacing, form a reticnlum, or meshwork. Towards the periphery of the tubercle the reticulum may become arranged in the form of a capsule as the age of the tubercle advances, and the reticular giant cell becomes eventually converted into fibrous tissue. The bacillus has disappeared, and the tubercle has healed. Giant cells cannot be relied upon to indicate tuberculosis. They are not always present in tubercu- losis, and they are not peculiar to tubercle, being found, for example, in actinomycosis. The only certain indication of tuberculosis is the pre- sence of the tubercle bacillus, which FIG. 163.— LA RGB OVAL GIANT CELL FROM TUBERCLE OF LUNG x 300 : a, Granular centre; b, nucleated periphery forming a mantle-like sheath ; c, processes from the same. 378 INFECTIVE DISEASES. can be revealed either by microscopical examination of the suspected tissue, or after inoculation in guinea-pigs. Bacillus Tuberculosis (Koch). — Rods, 2 to 4 //, and occa- sionally 8 /A long, very thin, and rounded at the ends. They are straight or curved, and frequently beaded, and occur singly, in pairs, or in bundles ; there are also involution forms and short branched threads. Spore-formation is observed in old cultures. They are non- motile. Tn the interior of giant cells they are often accompanied by grains which exhibit the same colour reaction. The bacilli in tissue sections of bovine tuberculosis are shorter and less granular than those in human tubercular sputum, but in milk they are quite as long, and even longer, and very distinctly granular or beaded, and are thus brought much closer, morpho- logically, to the bacilli in human sputum. Speaking generally, however, the average length of the human bacilli is greater than the average length of the bacilli in cow's milk, but the longest of the bovine bacilli cannot be distinguished in length from the longest human bacilli. There are, however, exceptional cases, for in some preparations of pus from human lungs the bacilli are remarkable, not only for their thinness, and their uniformly beaded character, but more particularly for their extraordinary length. They should be compared with other preparations, in which the bacilli, though in human sputum, are sometimes much more distinctly rod-shaped, much shorter and thicker, with complete absence of any beaded appearance. Neither length nor granularity is a characteristic sufficient to denote any specific difference between human and bovine bacilli. The author has examined minutely the bacilli in tuberculosis of other animals, such as the horse, pig, and cat ; and of birds — the fowl, guinea-fowl, pheasant, and ostrich. Here, again, minute morphological differences can be observed. For example, in many cases in fowls the bacilli are conspicuously short and straight. In the liver and lungs of an ostrich, packets of short rod-forms are found, while in other parts of the same sections the bacilli attain a very great length. Many of the long, sinuous forms exhibit a peculiar terminal enlargement. There are also short rods with a similar appearance, and free ovoid bodies, singly and in groups, which, from their connection with the bacilli, and their sharply defined outline in the free state, are similar to spores in old cultures. Thus, morphological differences are found under different circum- stances, and within limits the morphology of the tubercle bacillus varies with its environment. Koch first succeeded in cultivating the bacillus by employing DESCRIPTION OF PLATE XI. Bacillus tuberculosis. The figures in this plate represent the bacilli of tuberculosis in different animals, examined under the same conditions of amplifica- tion and illumination. x 1200. Lamp-light illumination. FlG. 1. — Bacilli in pus from the wall of a human tubenmlar cavity. In this specimen the bacilli are shorter than those in tubercular sputum, and are very markedly beaded. FIG. 2. — Bacilli in pus from a tubercular cavity from another case in man. They are present in the preparation in enormous numbers. The proto- plasm occupies almost the whole of the sheath, and the bacilli are strikingly thin and long. FlG. 3. — Bacilli in sputum from an advanced case of phthisis, showing the ordinary appearance of bacilli in sputum ; some beaded, others stained in their entirety ; occurring both singly and in pairs, and in groups resembling Chinese letters. FlG. 4. — Bacilli in a section from the lung in a case of tuberculosis in man. The bacilli in human tuberculosis are found in, and between, the tissue cells ; and sometimes, as in equine and bovine tuberculosis, in the interior of giant cells, but not so commonly. FIG. 5. — From a cover-glass preparation of the deposit in a sample of milk from a tubercular cow. The bacilli were longer than the average length of bacilli in bovine tissue sections, and many were markedly beaded. FIG. 6. — From a section of the brain in a case of tubercular meningitis in a calf, showing a giant cell containing bacilli with the characters usually found in sections of bovine tuberculosis. FIG. 7. — From a section of the liver of a pig with tubercle bacilli at the margin of a caseous nodule. FIG. 8. — From a cover-glass preparation of a crushed caseous mesenteric gland from a rabbit infected by ingestiori of milk from a cow with tuberculosis of the udder. FlG. 9.— From a section of lung in a case of equine tuberculosis, showing a giant cell crowded with tubercle bacilli. FIG. 10. — From a section of lung from a case of tuberculosis in the cat, with very numerous tubercle bacilli. FIG. 11. — From a cover-glass preparation of a crushed caseous nodule from the liver of a fowl, with masses of bacilli. These are for the most part short, straight rods ; but other forms, varying from long rods to mere granules, are also found. FIG. 12. — From sections of the liver and of the lung in a case of tubercu- losis of a Khea. Isolated bacilli are found, as well as bacilli packed in large cells, colonies of sinuous bacilli, and very long forms with terminal spore-like bodies and free oval grains. The preparations from which these figures were drawn were all stained by the Ziehl-Neelsen method, with the exception of the first, which was stained by Ehrlich's method. Plate XL. \ ,'. A \ "Fit* 3 Fig 5. v- Pig 6 X Pig 7. \ \ -7 \ a ' x^ ^^ \vj' f i * •/ ' BACILLUS TUBERCULOSIS. •vTlBKA/fp1 OF THB "UNIVERSITY Or TUBERCULOSIS. 379 blood serum. Solid blood serum, with or without the addition of gelatine, was employed, and the cultures incubated at 37° C. The growth takes place very slowly, and only between the temperatures of 30° C. and 41° C. In about eight or ten days the growth appears as little whitish or yellowish scales and grains. The bacillus can also be cultivated in a glass capsule, on blood serum, and the appearances of the growth studied under the microscope. The scales or pellicles were described by Koch as made up of colonies of a perfectly characteristic appearance, which may be still further studied by making a cover-glass impression. They are then seen to be composed of bacilli, arranged more or less with their long axis corresponding with that of the colony itself, and with an appreciable interval between the individual bacilli. The colonies themselves appear as fine curved lines, the smallest being mostly S~snaPe(i- Longer colonies have serpentine twistings and bendings, which often recall the curves of fancy lettering. The ends of the lines run to sharp points, but the middle of the growth is spindle -formed. The youngest colonies are extremely delicate and narrow, but the older colonies increase in size, are thicker across, and, FlG- 164. - BACILLUS TUBERCULOSIS, FROM TUBERCULAR SPUTUM, x 2500. From Photo- blending with each other, graphs gradually obliterate the characteristic appearances ; a lamellated growth results, which increases, and gives the appearance to the naked eye of the scale or pellicle already described. The blood serum is not liquefied unless putrefactive bacteria contaminate the culture. A fresh tube can be inoculated with one of the little scales, and a new generation started. The scales gradually increase in size, and consist entirely of bacilli. In about three to four weeks the cultivation ceases to increase, and it is then necessary to inoculate a fresh tube. In liquid blood serum a film forms on the surface of the liquid, and is easily broken by agitation. In neutralised broth there is very little indication of success. When a triturated culture is added to the broth, a granular, sandy, whitish deposit collects at the bottom of the vessel, with indications of an increase in amount. Koch also tried nutrient agar-agar, which did not prove to be at 380 INFECTIVE DISEASES. BACILLUS TUBERCULOSIS. FIG. 165. — PURE-CULTIVATIONS ON GLYCERINE-AGAR FROM HUMAN TUBERCULAR SPUTUM, a, After six months' growth. (Fifth sub-culture.) b and c, After ten months' growth. (Fourth sub-cultures.) TUBERCULOSIS. 381 all a favourable medium. Some increase took place, but there was no continuous growth over the inoculated area. Glycerine Agar-agar. — Nocard and Roux were among those who worked at the subject and confirmed Koch's observations, Nocard attempted to get cultures of avian tuberculosis on blood serum to which peptone, salt, and cane sugar had been added. The results were more success- ful than with ordinary serum. But he encountered a difficulty in the rapid drying of the surface of the medium, which rendered the tubes unfit for use. It occurred to Nocard and Roux to obviate this by the addition of a hygroscopic agent, and accord- ingly they added sterilised glycerine. The result, which far exceeded their expectation, evidently was not solely attributable to the prevention of desiccation. Following up their discovery, and being anxious to find a medium more easily prepared than blood serum, they added 6 to 8 per cent, of glycerine to ordinary nutrient agar-agar. The bacillus grew so abundantly in this mixture that a culture in fifteen days equalled in extent a culture on blood serum which was several weeks old. The bacillus was found to grow abundantly in veal broth, to which glycerine had been added in the proportion of 5 per cent., the bottom of the flask being covered in about three weeks with a flocculent deposit, having some resemblance to anthrax cultivations in liquid media. In beef broth, chicken broth, and in Cohn's liquid, cultures were obtained after the addition of glycerine. Description of Cultivations on Glycerine Agar-agar. — The cultivations on the sloping surface of obliquely solidified glycerine agar- agar begin to appear in from four to six days as very minute white colonies. These steadily increase in size, and either look moist and FIG. 166.— PURE-CULTI- VATION IN GLYCERINE AGAR-AGAR, after ten months' growth. FIG. 167. -PURE-CULTI- VATION IN GLYCERINE AGAR-AGAR, — A SUB- CULTURE FROM A PURE- CULTURE IN GLYCERINE- MILK. In two months. 382 INFECTIVE DISEASES. smooth, or, even at this early stage, appear dry and crinkled. According to the number of bacilli inoculated, the colonies will either remain isolated or coalesce and form a more or less continuous film. If the nutrient agar-agar has only recently been prepared, there is usually a quantity of liquid present, and the bacillus forms a white coating over the inoculated area and beyond it. The appearances are much more characteristic when this medium is, comparatively speaking, dry. A semi-transparent membranous growth develops, thickens, and assumes a characteristic lichenous appearance. Such a culture, examined with a pocket lens, resembles a model in wax in miniature of the folds of the gastric mucous membrane. In about six weeks to two months the culture has fully developed. In old cultures, especially when the individual colonies remain isolated, the appearance is very characteristic. Some cul- tures in appearance closely resemble cultivations on blood serum. The consistency of the growth depends upon the character of the soil and the age of the culture. When the medium is moist the growth is moist and viscous, but more often it is distinctly tallowy, and in old and dry cultures scaly and friable. Cultivations in Glycerine Broth. — In a few days minute flakes are visible, which steadily increase in size, and subside to the bottom, of the flask, forming in time a very copious deposit. On shaking the flask, this deposit, which is extremely tenacious, rises in stringy masses, and gives an appearance which is more or less character- istic. If the flask is left undisturbed, a delicate veil-like film forms over the surface, which can be readily broken up by gentle agitation, forming flakes which gradually sink in the liquid. If undisturbed for several weeks this film increases in thickness, is irregularly fissured, and has more the appearance of masses of tallow floating on the surface. The growth also may be seen to extend up the side of the flask above the liquid. Pasteur or Erlenmeyer flasks can be employed for these cultures. Solidified egg-albumin added to the glycerine broth seems to increase the amount of growth, which clings to the albumin, and waves to and fro in the liquid when the flask is gently shaken. The author has confirmed the observation of Nbcard and Roux, that sub-cultures from glycerine agar-agar, or from glycerine broth, will give cultures in ordinary broth without glycerine. Ordinary broth with egg-albumin, and without glycerine, will also give a good growth when inoculated from previous sub cultures, although the attempt to produce primary cultures in these media has hitherto failed. Cultivations in Glycerine- Milk, and other Media. — In milk the TUBERCULOSIS. 383 author found there was only a slight increase in the number of bacilli inoculated, but milk with glycerine in the proportion of 5 per cent, forms a more favourable medium. The author has also cultivated the bacillus on sterilised urine and glycerine, and ordinary nutrient gelatine with 5 per cent, of glycerine. On potato the growth of the bacillus is extremely slow. Beevor succeeded in obtaining cultures at the ordinary temperature of the room. Examination of Cultivations. — To examine the bacilli in these various preparations the author prefers to use Neelsen's method, floating the cover-glasses for from five to ten minutes on warm carbolised fuchsine, and passing them through dilute sulphuric acid. In some cultures the bacilli are shorter and thicker than is commonly observed in human sputum, and they are for the most part without the beaded appearance. In old cultures on glycerine agar-agar the number of granular or beaded bacilli increases, and there are also numerous peculiar forms. There are bacilli, sometimes two or three times the length of an ordinary bacillus, provided with a club-shaped enlargement at one or both extremities, and in rare cases with lateral branches. They are no doubt identical with the bacilli with swollen extremities and the branched forms observed by Nocard and Roux. In milk the appearance is very striking, many bacilli attaining in old cultures a great length, and all are more uniformly beaded than in any other cultivations. Staining preparations by the method of Grain in all cases exaggerate this appearance. The important part played by the environment is shown by the morphological differences observed in artificial cultivation under varying conditions, and by the fact that by successive cultivation the bacillus can be educated to grow upon a medium which is un- suitable for obtaining primary cultures. Impression preparations of the growth of the bacillus on the surface of glycerine agar-agar in capsules show a tendency to the formation of serpentine colonies, composed of bundles of more or less parallel bacilli. Spore-formation. — In old cultivations true spore-formation can readily be observed, both in stained and unstained preparations. In the latter case they are recognised in the form of one or two highly refractive bodies in individual bacilli. Toxic Products of Cultures. — The poisonous substances found in cultures, and the composition and use of tuberculin, have already been described (p. 43). Inoculation Experiments. — A relatively small portion of a culti- /vT' B R * ^~\^ f OF THK I UNIVERSITY ) ^w. r> _ • 384 INFECTIVE DISEASES. vation inoculated into the subcutaneous tissue, into the peritoneal or pleural cavities, into the anterior chamber of the eye, or directly into the blood stream, produces after three or more weeks artificial tuberculosis in guinea-pigs and rabbits. Dogs and cats can also be infected by experimental inoculation. When a trace of tubercular virus is inserted subcutaneously in the thigh of a guinea-pig, in about a week or ten days a chain of enlarged glands will be easily felt in the vicinity of the seat of inoculation. This affords an unfailing test, which can be applied when there is difficulty in ascertaining by the microscope the presence of the bacilli in the material under examination. It also affords a valuable method for testing the effects of antiseptics on tubercular virus. The appearances observed at the autopsy are swollen lymphatic glands, in the neighbourhood of the inoculation, followed by softening and abscess ; enlargement of the spleen and liver, with formation of caseous tubercles ; and tubercular deposits in the lungs, bronchial glands, and peritoneum. After inoculation of the eye, grey tubercles appear on the iris, and undergo enlargement and caseation, followed by tuberculosis of the eyeball and organs generally. The bacilli appear to be the direct cause of tuberculosis, and the presence of the bacillus in the sputum of patients is a distinctive sign of the existence of this disease. The detection of the bacillus has, consequently, become a test which is constantly applied. The bacilli are found in all tubercular growths of man, monkeys, cattle (Perlsuckt), birds, and many other animals, and in cases of artificial tuberculosis, in rabbits, guinea-pigs, cats, etc. In man the bacillus can be detected in the tissues, in the sputum, in the blood, and in the urine. Tuberculosis may also be produced by inhalation and feeding experiments. The channels of infection in man are also most probably the pulmonary and intestinal mucous membranes. The possibility of inoculation of skin wounds is open to doubt. The bacilli or their spores are inhaled from the air, or taken in with food. Morphologically identical bacilli have also been observed, but very sparsely, in sections of lupus. METHODS OF EXAMINING THE TUBERCLE BACILLUS. Numerous methods have been recommended for examining the tubercle bacillus. A few of these will be described, as many are only of historical interest. TUBERCULOSIS. 385 The Ziehl-Neelsen method is preferred by the author both for sections and cover-glass preparations. KocK* ni'iijhinl method. — Cover-glass preparations or sections are laid in Koch's solution (No. 23, r) for twenty-four hours, or for one hour if the solution is warmed to 40° C. Rinse in water ; immerse in a watery solution of vesuvin for two minutes ; rinse again in water, and examine ; or, after rinsing in water, treat with alcohol, clove-oil, and Canada balsam. rJn-HcTC* method. — Cover-glass preparations are allowed to float in a watch-glass, containing a solution of gentian-violet or fuchsine, added to aniline water. A saturated alcoholic solution of the dye is added till precipitation commences (10 cc. aniline water, and 10 to 20 drops of the colour solution). The cover-glasses are left in the solution for about half an hour ; then washed for a few seconds in strong nitric acid (one part commercial nitric acid to two of distilled water), and rinsed in distilled water. After-stain with vesuvin or methylene-blue, rinse in water, dry and preserve in Canada balsam. . Ehrlich-Koch method. Saturated alcoholic solution of methyl- violet or fuchsine 11 Aniline water ........ 100 Absolute alcohol ........ 10 Preparations are left for twelve hours in this solution (colouring of the cover-glass preparations can be expedited by warming the solution). Treat the preparations with (1 to 3) solution of nitric acid a few seconds. Wash in alcohol (GO per cent.) for a few minutes (cover-glass prepara- tions need only be rinsed a few times). After-stain with diluted solution of vesuvin or methylene-blue for a few minutes. Wash again in 60 per cent, alcohol, dehydrate in absolute alcohol. Clear with cedar-oil, mount in Canada balsam. Rindfteisch's method. — Prepare a solution composed of Saturated alcoholic solution of fuchsine . . 10 drops Aniline water ....... 2 drams. Pour it into a watch-glass, and float the cover-glass ; warm the watch- glass over a spirit-lamp until steam rises. Remove it from the flame, and set it aside for five minutes. Take out the cover-glass, and transfer it for a few seconds to acidulated alcohol (two drops of nitric acid in a watch-glass full of alcohol). Wash in distilled water, dry, and preserve in balsam. After-stain, if necessary, with Bismarck-brown, or methylene- blue. Gibbet n«t //<><}.— Cover-glass preparations are placed in Gibbes' double- staining solution which has been warmed in a test-tube, and, as soon as steam rises, poured into a watch-glass. They are allowed to remain for five minutes, and then are washed in methylated spirit till no more colour comes away, dried in the air or over a spirit-lamp, and mounted in Canada balsam, Jf the solution is used without warming, the cover-glasses 386 INFECTIVE DISEASES. must be left in it for an hour. Sections are treated on the same principles, but must be left in the solution for several hours. The crumpling of the sections by the action of nitric acid is avoided. Bawngdrten's method. — Cover-glass preparations of sputum are made as already described, and immersed in a very dilute solution of potash (1 to 2 drops of a 33 per cent, solution of potash in a watch-glass of dis- tilled water). The cover-glass is pressed down on a slide, and examined with a high power. The bacilli can be thus examined in the unstained condition, and to avoid any mistake from confusion with other species, the cover-glass can be removed, dried, passed through the flame, and stained with a drop of an aqueous solution of fuchsine, or gentian-violet. The putrefactive bacteria are stained, but the tubercle bacilli remain absolutely colourless. Baumgarten's new method. — A solution is prepared as follows : Drop 4 to 5 drops of concentrated alcoholic methyl-violet solution into a small watch-glass full of water, (a) Stain the sections in this solution, wash them in water, and decolorise in absolute alcohol (five to ten minutes) ; or, before treating with alcohol, immerse the sections for five minutes in a half- saturated solution of carbonate of potash. Pass through clove-oil, and mount in a mixture of Canada balsam, free from chloroform, and clove-oil (equal parts). The object of this process is to differentiate the tubercle bacilli from chance bacteria, inasmuch as the tubercle bacilli are gradually decolorised by the clove-oil. (6) Sections stained in the above solution are placed for five minutes in alcohol, and then in a concentrated solution of Bismarck-brown in 1 per cent, solution of acetic acid. The after-treatment may be conducted as already described. Ziehl-Neelsen method. — Cover-glass preparations may be quickly stained in Neelsen's solution warmed in a watch-glass till steam rises. Sections are left for from five to ten minutes in the solution, and then washed in a watery solution of sulphuric acid (25 per cent.), rinsed in distilled water, and immersed in methylene-blue solution. After two or three minutes they are passed through alcohol and oil of cloves, and mounted in Canada balsam. FranlceVs method. — Sputum preparations are rapidly double-stained by the following method : Prepare a solution by adding concentrated alcoholic methyl-violet or fuchsine solution, drop by drop, till opalescence arises, to 5 ccm. of aniline- water heated to 100° C. Float the prepared cover-glasses two minutes in the warmed solution. The process of after- staining and decolorisation is effected by placing the preparation for one to two minutes in one of the following solutions : for fuchsine-stained preparations, a saturated solution of methylene-blue in a mixture of Alcohol 50 Distilled water .30 Nitric acid 20 which is filtered before use ; for preparations stained in methyl- violet, a saturated solution of vesuvin may be used in Alcohol .70 Nitric acid ... 30 TUBERCULOSIS. 387 Ehrlich's Method and Eosin. — The author has found that after sections have been stained with methyl-violet and Bismarck-brown by Ehrlich's method, as described by Koch, they may with advantage be immersed in a weak alcoholic solution of eosin, then rinsed in clean absolute alcohol, clarified with clove-oil, and mounted in Canada balsam. The giant cells are then stained pink, while their nuclei are brown, and the bacilli blue. TUBERCULOSIS IN MAN. The disease manifests itself in various forms in man, and most frequently in the lungs, producing phthisis or consumption. The sputum contains the bacilli in large numbers, and is extremely virulent. Scrofula and lupus are forms of tuberculosis ; they are FIG. 168.— SECTION THKOUGH A LUPUS NODULE OF THE NOSK. probably produced by an attenuated variety of the tubercle bacillus. Lupus can be distinguished from tuberculosis of the skin ; and scrofulous lymphatic glands are distinguished from tubercular glands by the tendency of the latter to produce generalised tuberculosis. This difference in the intensity of the virus in the two cases, Lingard illustrated by the effect upon inoculated guinea-pigs. Cavities in the lungs are often thickly lined with bacilli. They are present in great numbers in the caseous matter, though in equine and bovine tuberculosis this is not the case. Whether the disease in man is contagious is an open question, though numerous cases of supposed communication between husband and wife, brothers and sisters, have been reported, and Ransome 388 INFECTIVE DISEASES. showed that tubercle bacilli were present in the breath in phthisis. On the other hand, the experience in consumption hospitals does not FIG. 169.— TUBERCULAR ULCERATION IOP MUCOSA OF HUMAN ILEUM. Between the ulcers there are tubercular lymph-follicles (HAMILTON). support this view, there being no evidence of the communication of the disease to nurses and hospital attendants, TUBERCULOSIS. 389 TUBERCULOSIS OF CATTLE. In cattle the disease may occur as the result of inhaling bacilli, or of ingestion with food. It is very frequently found in the lungs ; and calves may be infected by milk from cows with tubercular udders. Calves may also suffer from congenital tuberculosis, the bacilli having been transmitted from the mother during gestation. Breeding in-and-in, over-production of milk, and confinement with insanitary surroundings, predispose to tuberculosis. The disease is known in Germany as " Perlsucht " ; and in this country the lesions on the pleura are known as "grapes," and the animals themselves are commonly called " wasters." The disease may also exist in the lungs or in other organs, in a limited form, without any indication of ill health. In such cases the disease can be detected by injection of tuberculin, a marked rise of temperature occurring in t u b e r c u 1 a r animals. In advanced cases, the symptoms commonly observed are cough, difficulty in breath- ing, staring coat, wasting, and diarrhoea ; and if the udder is infected, nodules in the gland, arid thin bluish milk. In the lungs, after slaughter, a few small cheesy tubercles may be found in animals apparently in perfect health and in prime condition for the market. In advanced cases, the lungs on section show large yellow masses, containing calcified matter, and the bronchi may be full of yellowish pasty contents. The disease will be found to involve the bronchial glands. The serous membrane may be covered with little warts or grape-like masses. The lymphatic glands may be enlarged to an enormous size. Tubercular ulceration of the intestine is sometimes found, but not commonly. In tubercular disease in the udder, a painless swelling is found which may affect one or more quarters of the gland. Transmission of Tuberculosis from Man to Cattle. — It is FIG. 170. —SECTION OF LUPUS OF THE SKIN, x 700. Giant cell containing a tubercle bacillus (FLUGGE). 390 INFECTIVE DISEASES. for obvious reasons impossible to ascertain by experiment whether tuberculosis can be transmitted from cows to man by milk or other- wise ; but some light may be thrown upon this important question by ascertaining the result of inoculating bovines with human tuber- culosis. If calves can be infected with tuberculosis from a human source by inoculation or ingestion experiments, and especially if the effect of administering human and bovine tubercle to calves by these means is found to be the same, such experiments will not only serve to dispel any doubt there may be as to the identity of the two affections, but they will strengthen the hands of those FIG. 171.— TUBERCULOSIS OF PLEURA ; ".GRAPE- DISEASE." who insist upon the necessity of more thorough inspection of dairy cows, and of power to deal with tubercular animals. Inoculation of a Calf with Human Tubercular Sputum. — The author obtained sputum containing numerous bacilli from an advanced case of phthisis. The sputum was shaken up with sterilised salt solution and injected into the peritoneal cavity. A few weeks afterwards the^calf showed signs of illness. The animal looked dull, did not feed well, had a slight cough, and showed less inclination to move about than usual. These symptoms gradually increased, and death occurred forty-two days after inoculation. Extensive lesions TUBERCULOSIS. 391 were discovered at the post-mortem examination. The mesentery was adherent to the abdominal wall, at the seat of the inoculation, and to the rumen ; the liver was adherent to the diaphragm. There was extensive tubercular deposit at the seat of inoculation, and an abscess the size of a walnut. Extending over the mesentery from this point there were hundreds of wartlike, fleshy, new growths, some quite irregular in form, others spherical or button-shaped. There were similar deposits on the under surface of the liver, on the spleen, in the gastro-splenic omentum, and on the peritoneal surface of the diaphragm. The spleen was adherent to the rumen, and on dissecting away the adhesions another abscess was opened. The lungs were congested and the pleurae thickened. On microscopical examination of sections extremely minute tubercles were found to be disseminated throughout the whole of the substance of the lungs and liver, and tubercle bacilli were found in these and in the peritoneal deposits. The abscesses contained Streptococcus pyogenes. The calf died of pyaemia, but sufficient time had elapsed for marked local infection leading to generalised miliary tuberculosis. TUBERCULOSIS IN RELATION TO THE PUBLIC MILK SUPPLY. There is not the slightest doubt that when the udder is involved the milk is highly virulent to the lower animals, and presumably is, therefore, dangerous to man. The virulence of the milk was first insisted upon by Klencke in 1846, and confirmed by Gerlach in 1869, and later, by others. This subject was again brought forward with the discovery of the tubercle bacillus, and the demonstration of its existence in the milk in certain cases of bovine tuberculosis. Koch pointed out that the milk only contained bacilli, and was only infective, when the udder itself was tubercular. By this he explained the contradictory results obtained by various experimenters with milk from cows un- doubtedly suffering from " Perlsucht." Koch considered that positive effects were obtained with milk when it happened to contain tubercle bacilli, and negative with milk from which they were absent. Bang in a number of cases verified the presence of tubercle bacilli in milk, and, owing to the contradictory results of previous investigations, repeated the ingestion experiments. The milk was found to be virulent both to pigs and rabbits. In this country Woodhead and M'Fadyean tested milk for tubercle bacilli. They examined six hundred cows in the Edinburgh dairies, and found thirty-seven suffering from mammitis, but in only 392 INFECTIVE DISEASES. six were they able to demonstrate the presence of tubercle bacilli in the milk, and then only in small numbers. Hirschberger found in twenty cases of tuberculosis in cattle that the milk of eleven was virulent to guinea-pigs. Three cows out of nine in which the disease was restricted to the lungs gave infected milk. On the other hand, Nocard inoculated milk from eleven tuberculous cows, of which only one had diseased udder, and only this one gave infective milk. Bang injected rabbits with milk from twenty-one cases of tuberculosis, with the udders apparently normal, and the milk was virulent in two. The author had two cases of udder tuberculosis under observation, and as no experiments had at the time been made in this country with milk known to contain tubercle bacilli, it was decided to study the effect on rabbits, and test the results obtained by Bang. These cases were both interesting and instructive, and may be referred to in detail. One was a case of advanced general tuberculosis. There was extreme emaciation, general apathy, and a peculiar dull expression of countenance. The skin was dry and harsh, the coat staring, and there was loss of hair in patches about the face and neck. There was dulness on percussion over a large area of the thorax, and the respirations were increased in rapidity. There was also occasional cough and some diarrhoea. But the most interesting condition was observed on examination of the udder. The gland was swollen, especially posteriorly, and distinct induration could be felt on examination. The deposit appeared to be more or less limited to the posterior quarters. The cow evinced no pain during the examination of the udder, not even on the application of firm pressure. The author took samples in test-tubes of the milk from all four teats ; when freshly drawn, it differed noticeably from the normal secretion. It was a thin, watery, turbid fluid with whitish flakes in suspension, but it was not gelatinous or muco-purulent in character, and was free from any markedly yellow colour. After being set aside in the laboratory for some hours it separated into a layer of cream and a turbid liquid of a yellowish tint, while at the bottom of the test-tube there was a whitish flocculent deposit, especially in the samples from the posterior quarters. There were tubercle bacilli both in the cream and in the deposit. In the cream they were only present in small numbers, and were detected, therefore, only after careful search. But in the deposit they were readily found, as in a cover-glass preparation there were sometimes four or five in the field of the microscope. The method adopted for the examination of this deposit was as follows : The whole of the liquid in the test-tube was carefully poured off, and a trace of the sediment spread out on a cover-glass. This was allowed to dry, and passed through the flame, and stained in hot Ziehl- Neelsen solution in the usual manner. TUBERCULOSIS. 393 The other cow was also a case of general tuberculosis, and presented somewhat similar lesions of the udder. The induration of the gland was readily detected, and examination of the milk showed, as in the previous case, the presence of tubercle bacilli. It will be observed that in neither of these cases was the disease limited to the udder ; in both the implication of the gland was part of general tuberculosis. FIG. 172.— TUBERCULAR ULCERATION OF THE INTESTINE OF A Cow. The first cow was killed, and the following lesions were found at the post-mortem examination. T/nn'dx. — The lungs and bronchial glands were extensively invaded with tubercular deposit. The glands were greatly enlarged and densely fibrous, in many cases with central, stone-like masses, grating on section against the edge of the knife. In the lung there was every stage, from the early deposit to purulent cavities, cheesy masses, and calcified dtbrt*. Abdomen. — There were a few caseous nodules in the liver, but none in the spleen. The mesenteric glands formed an almost continuous chain 394 INFECTIVE DISEASES. of large tumours, mostly with central cretification. Tubercular deposit in the intestines could be recognised from the outside, and on laying them open the mucous membrane was found to be studded with tuber- cular ulcers. These ulcers were most numerous in the large intestine, and varied in size from a sixpence to a florin. Some were circular, others slightly irregular in form, and others again distinctly oval. In the latter case they were generally situated with their long diameter transversely. The base of the ulcer involved the muscular coat, and was irregularly radiated. The margin was broad, and elevated above the general surface, producing a ring-like appearance. Mammary Gland. — The udder was infiltrated throughout with tuber- cular new growth, but the invasion was most marked in the posterior quarters. There was apparently very little tendency to caseation. Microscopical Examination of the Udder. — In order to study the histo- logical characters of the gland, and the distribution of the bacilli, sections were stained with logwood and rubin, and others again with fuchsine and methylene-blue. The tubercular new growth consisted of the usual histological elements, round cells, epithelioid cells, and giant cells. Healthy lobules here and there were sharply marked off from those in which the growth was compressing and obliterating the alveoli in its progress. Bacilli were present in the giant cells, and also distributed in vast numbers throughout the tubercular tissue generally. Bacilli were found in epithelioid cells close to the alveolus, and also between the cells lining the alveoli. In parts also the new growth had involved the milk ducts, and therefore it was easy to account for the presence of the bacilli in the milk. The bacilli were found in considerable numbers also in sections of the intestinal ulcers. EXPERIMENTAL INFECTION OF BABBITS. Ingestion. — A rabbit received the contents of a test-tube which had been filled with milk from one of the posterior teats, mixed with a small quantity of bran. In four weeks there was commencing emaciation ; later, diarrhoea set in, and death occurred exactly fifty- eight days after administration of the milk. At the post-mortem examination the mesenteric glands were found to be much enlarged and caseous. A cover-glass preparation from a crushed gland revealed numerous tubercle bacilli. On opening the intestines there was a patch of ulceration, showing the point of access of the bacilli. The intestinal ulceration was a reproduction, to a certain extent, of the condition in the cow which had been the source of the virus. Subcutaneous Injection. — A second rabbit was injected under the skin of the back by means of a capillary pipette with about ten drops of milk, including some of the deposit from the bottom of the test-tube. The sample of milk had in this case also been taken from DESCRIPTION OF PLATE XII. Tubercular Mammitis. FIG. .1.— From a section of the udder of a milch cow. The tubercular deposit is seen to invade the lobules of the gland. Lobules comparatively healthy are marked off, more or less sharply, from the diseased ones in which the new growth in its progress compresses and obliterates the alveoli. Stained by the Ziehl-Neelsen method and with methylene-blue. x 50. FIG. 2.— Part of the same preparation. On the right of the section part of a healthy lobule is seen. On the left a lobule is invaded by tubercular new growth composed of round cells, epithelioid cells and typical giant cells. Tubercle bacilli can be seen both singly and collected in groups. They are found in and between the cells, and in the interior of giant cells. Bacilli may be seen between the cells lining an alveolus and projecting into its lumen, x 800. %.,J — •*:.'.•?">•? ,'- :';•-. |^1>* '-;.. v^r--;-.....^/ ./ \Vs._. ,-/ ~Y>: Ftfl. Fig 2. TUBERCULAR MAMMITIS E.Jf.CrooJwluinJe/ecit TUBERCULOSIS. 395 one of the posterior teats. The rabbit was placed in a separate hutch, and death from general tuberculosis occurred ninety- two days after inoculation. The diaphragm and mesentery were studded with tubercles the size of a pin's head. The kidneys superficially showed whitish rounded nodules projecting above the surface. These were found on section to be continuous, with wedge-shaped deposits in the sub- stance of the kidney. The lungs presented a very striking appear- ance, being, in short, a mass of tubercular deposit ; and the bronchial and tracheal glands were similarly affected. In sections of the kidney and lung the bacilli were present, but they were distributed irregularly ; in one part of a section it was difficult to detect a single bacillus, in other parts they were present in large numbers. The milk from the two cows, previously to their coming under observation, had been mixed with the general supply of a dairy. There is indeed ample evidence that, both in this and in other countries, the milk of tuberculous animals finds its way into the market. The question which naturally arises is the possi- bility of any manifestation of tuber- culosis in man, arising from the consumption of unboiled milk con- taining tubercle bacilli. We must admit that there is no direct evidence of the transmission of tuberculosis by milk from cow to man ; but this may arise from the difficulty in tracing such a source of infection, owing to the long time which elapses before symptoms manifest themselves in man. Yet, if milk be a source of infection, we should naturally expect that primary tuberculosis of the intestine would be by no means an uncommon manifestation of the disease; and this in the adult is not in accordance with clinical experience. Such an argument would tend to centra-indicate danger to adults; but, on the other hand, the possible danger to children has been rightly insisted upon by the earliest writers on this subject. Woodhead has recently stated that, from his experi- ence in two large hospitals, he has been much struck by the fact that, in children who had died from other diseases during the course of tubercular disease of the abdominal glands, there was frequently FIG. 173.— TUBERCULAR ULCERA- TION OF THE INTESTINE OF A RABBIT. 396 INFECTIVE DISEASES. not any trace of tubercular disease in other parts ; thus pointing to the intestine as the channel by which the bacillus made its way into the body. Woodhead also remarks that in a large number of cases FIG. 174.— TUBERCULOSIS OF THE LUNGS. From a photograph of the lungs of a rabbit which had been injected sub- cutaneously with about ten drops of milk, including in suspension a small quantity of the deposit at the bottom of a sample of milk from a cow with tuberculosis of the udder. Death occurred from general tuberculosis ninety-two days afterwards. The appearance of the lungs was very striking. They were almost completely composed of tubercular deposit. The bronchial glands, as well as the tracheal, of which one is seen in the photograph, were also enlarged and caseous. There were tubercular deposits in the kidneys and other organs, and also at the seat of inoculation. of general tuberculosis, where the possibility of infection by the pulmonary passages was evidently excluded, the tubercular process TUBERCULOSIS. 397 appeared to have invaded the body by the intestinal canal. These facts, taken in connection with the occasional existence of tubercle bacilli in milk, went far to prove, in his opinion, that milk was a source of tubercular infection, especially to young children. From his own experiments and observations the author has drawn the following conclusions : — 1. Cows with tuberculosis of the udder are to be found in dairies in this country. 2. The milk of these cows is, as a rule, mixed with the general supply. 3. The milk in cases of udder tuberculosis contains tubercle bacilli. 4. Rabbits inoculated with, or fed upon, milk containing tubercle bacilli contract tuberculosis. 5. Direct evidence of transmission of tuberculosis by milk to man is wanting, but from the effect of such milk on the lower animals it is reasonable to conclude, in the present state of our know- ledge, that there may be danger in using the milk of cows with tubercular udders, and therefore strict inspection of dairies should be enforced ; and boiling of milk before use will, as a rule, be a wise, if not absolutely a necessary precaution. Bellinger has shown that the virulence of cow's milk is reduced by dilution with water in the proportion of 1 in 40 and even of 1 in 100, and that therefore there would be much less danger in consuming tubercular milk which had been mixed with the milk of healthy cows, than there would be in taking it direct from the infected cow. This is a matter of scientific interest ; but it would be no justification for a dairyman to mix the milk of a tubercular cow with milk of cows known to be healthy. The milk of cows suffering from tuberculosis should undoubtedly be rejected. TUBERCULOSIS AND THE PUBLIC MEAT SUPPLY. The question of the advisability of allowing the flesh of tuber- cular animals to be sold for food, especially when the disease exists in a very small degree, is a vexed one. Numerous experiments have been made upon the infectivity of the flesh of tubercular animals. Kastner inoculated the juice expressed from the flesh of tubercular cows. Sixteen guinea-pigs were unaffected after injection 398 INFECTIVE DISEASES. of 1 to 2 cc. into the peritoneal cavity. Nocard injected ten to twenty drops of the muscle juice of the hearts of tubercular cattle, in which the disease was well marked, and none of the guinea-pigs were infected. With juice of the muscles of the thigh derived from ten tubercular cows Nocard inoculated forty guinea-pigs, and one only showed signs of tubercle. Nocard concluded that if there was any danger in the flesh of tuberculous animals, it was the exception and not the rule. On the other hand, Chauveau and Arloing produced tuberculosis in two guinea-pigs out of ten inoculated with muscle juice from a tubercular steer. In 1890 a Royal Commission was appointed to investigate this subject, and the report was issued in 1895. Martin, on behalf of the Commission, tested the flesh of twenty-one tubercular cows. In two cases only was evidence obtained of the presence of the bacillus by inoculation of guinea-pigs. The flesh of eight cows affected with mild tuberculosis produced tubercle in one instance by inoculation, but the ingestion experiments were negative. The flesh of five cows severely affected with tubercle gave the disease in four cases, either by feeding or inoculation, but only one gave the disease both ways. Martin thought that some of the results were due to the butcher infecting the meat in the process of dressing the carcase, either by his hands or knives. Woodhead made a series of experiments to test the effects of roasting and boiling on the tubercular virus in meat. It was found that in boiling and roasting experiments, as ordinarily carried out in the kitchen, the temperature, however high it may be on the surface, seldom reaches 60° C. in the centre, except in the case of joints less than about six pounds in weight. Boiling and roasting were found insufficient to destroy tubercular virus enveloped in rolls of meat. The following were among the conclusions of the Commissioners : — We have obtained ample evidence that food derived from tuberculous animals can produce tuberculosis in healthy animals. The proportion of animals contracting tuberculosis after experimental use of such food is different in one and another class of animals ; both carnivora and herbivora are susceptible, and the proportion is high in pigs. In the absence of direct experiments on human subjects, we infer that man also can acquire tuberculosis, by feeding upon materials derived from tuber- culous food-animals. The actual amount of tuberculous disease among certain classes of food-animals is so large as to afford to man frequent occasions for contracting tuberculous disease through his food. As to the proportion of tuberculosis acquired by man, through his food or through other means, we can form no definite opinion, but we think it probable that an TUBERCULOSIS. 399 appreciable part of the tuberculosis that affects man is obtained through his food. The circumstances and conditions with regard to the tuberculosis in the food-animal which lead to the production of tuberculosis in man are, ultimately, the presence of active tuberculous matter in the food taken from the animal, and consumed by the man in a raw or insufficiently cooked state. Tuberculous disease is observed most frequently in cattle and in swine. It is found far more frequently in cattle (full grown) than in calves ; and with much greater frequency in cows kept in town cow- houses than in cattle bred for the express purpose of slaughter. Tuber- culous matter is but seldom found in the meat substance of the carcase ; it is principally found in the organs, membranes, and glands. There is reason to believe that tuberculous matter, when present in meat sold to the public, is more commonly due to the contamination of the surface of the meat with material derived from other diseased parts, than to disease of the meat itself. The same matter is found in the milk of cows when the udder has become invaded by tuberculous disease, and seldom or never when the udder is not diseased. Tuberculous matter in milk is exceptionally active in its operation upon animals fed either with the milk or with the dairy produce derived from it. No doubt the largest part of the tuberculosis which man obtains through his food is by means of milk containing tuberculous matter. Provided every part that is the seat of tuberculous matter can be avoided and destroyed, and provided care be taken to save from contami- nation by such matter the actual meat substance of a tuberculous animal, a great deal of meat from animals affected by tuberculosis may be eaten without risk to the consumer. Ordinary processes of cooking applied to meat which has got con- taminated on its surface are probably sufficient to destroy the harmful quality. They would not avail to render wholesome any piece of meat that contained tuberculous matter in its deeper parts. In regard to milk we are aware of the preference by English people for drinking cow's milk raw — a practice attended by danger, on account of possible contamination by pathogenic organisms. The boiling of milk, even for a moment, would probably be sufficient to remove the very dangerous quality of tuber- culous milk. TUBERCULOSIS IN EQUINES. Tuberculosis is not very common in the horse, but when it does occur, it is frequently mistaken for glanders. There may be miliary tuberculosis in the lungs, or nodules disseminated throughout the lungs, liver, spleen, and bones. In a number of cases investigated by Nocard, the disease commenced in the abdominal organs, and the affection of the lungs appeared to be secondary. The author has examined several cases of equine tuberculosis. In some cases 400 INFECTIVE DISEASES. the lungs were affected with the disease in a miliary form. The bacilli could not be distinguished from bacilli in sections of the bovine disease. Giant cells were extraordinarily numerous, and in many cases were densely packed with bacilli, so that they could be recog- nised en masse under a low power. The bacilli were also distributed in the tissue generally, but were much more numerous in the giant cells. TUBERCULOSIS IN DOGS. Peters described a case of tuberculosis in a pet dog, from eating sputum from a tubercular patient. This is said to be a not uncommon cause of canine tuberculosis. TUBERCULOSIS IN CATS. Nocard reported a case of tuberculosis in a cat from eating tuber- cular sputum. The abdominal organs were diseased. Bellinger has described two cases of miliary tuberculosis. M'Fadyean also has described a case. The bacilli are very plentiful in the lung. A minute examination of the individual micro-organisms by the author did not reveal any distinctive character. TUBERCULOSIS IN SWINE. The author examined the tubercular liver of a pig. The pig was about six months old, and after suffering from cough and emaciation, died. The liver had caseous nodules scattered throughout its substance, some the size of a pea, and others larger. Tubercle bacilli without distinctive characters were found on examination of sections ; but it was in some parts of a preparation difficult to detect any bacilli, and in other parts there were not more than five or six in the field of the microscope. Tuberculosis in swine is said to be very rare in America. TUBERCULOSIS IN BIRDS. Hens, guinea-fowls, turkeys, pheasants, and partridges, are sub- ject to tuberculosis, and ostriches and other birds kept in confinement may contract the disease. Tuberculosis in fowls appears to be introduced principally with the food, the disease occurring commonly in the intestines and liver. DESCRIPTION OF PLATE XIII. Tuberculosis in Swine. Section of liver of a pig with scattered tubercular nodules. Microscopica sections of the liver showed tubercle bacilli in very small numbers. Plate XIII C Vincent Broole3.Day *5o7i,Zit)i. TUBERCULOSIS. 401 The author has examined several cases of so-called spontaneous tuberculosis in fowls. Sections of the liver were in one case remark- able on account of the extraordinary invasion of the caseous deposits with bacilli. Cover-glass preparations had been made from the liver in the following way for diagnostic purposes : A tubercle was readily picked out on the point of a scalpel and crushed between two slides, and the cover-glass preparations stained with the Ziehl- Neelsen solution. The bacilli are for the most part very small. A few attain a considerable length, but the majority are in the form of small, straight rods, with many sizes intervening between these rods arid isolated granules. In July 1888 the author received from Mr. Bland Sutton the liver and lungs of a Rhea, which had died in the Zoological Gardens. The lung was infiltrated with caseous deposits, and there were scattered caseous nodules in the liver varying in size from a pea to a marble. The naked-eye appearance of a section of the liver through these nodules, at once recalled to mind the naked-eye appearance of the deposits in the pig's liver already described. But whereas in microscopical preparations of the pig's liver, bacilli were very scantily present, the sections of the lung and liver of the E-hea contained bacilli in such extraordinary numbers that, under a power of fifty diameters, the collections of bacilli could be recognised as red granular masses. These red masses under a high power were re- solved into dense colonies of bacilli. In their number and their distribution in the tissues, in their varying size, and in the extra- ordinary length of the longest forms, they presented very interesting points for observation. From the naked-eye appearance of the disease and the general microscopical characters, as well as the presence of bacilli agreeing in their staining reactions with the classical tubercle bacilli, the author had no hesitation in pronouncing the disease to be avian tuberculosis. Klein, who had examined a similar case, alluded to it in a description of leprosy; but this disease is unknown in the lower animals, and all attempts to infect them from man have been almost, if not entirely, negative. The bacilli in the Rhea are principally collected in the caseous parts, but they are also found in the tissue generally, and often collected in large cells. In size they vary to a marked extent. In the cells they often form compact masses of short bacilli, but in other parts, both in collections and singly, they attain a greater length than is observed in any other form of tuberculosis. Some of the bacilli present a vary interesting appearance. They are provided 26 402 • INFECTIVE DISEASES. terminally with a sharply defined ovoid body. There are also collections of short bacilli, many with these spore-like appearances. The author has also seen free ovoid forms, sometimes singly, some- times in groups. From their connection with the bacilli and their sharply defined outline they are very suggestive of spores. Johne examined the livers of a number of fowls accidentally infected by phthisical sputum. Nocard reported an outbreak in a poultry-yard where the man in charge had consumption. He also found the disease amongst fowls fed with the infected organs of tubercular cattle. Subcutaneous inoculation, and feeding of fowls with sputum or bovine virus, will produce the disease. Experimental inoculation of tubercular virus from different sources affords an illustration of the different pathogenic effects obtained by varieties of the same species of bacillus. The bacillus of fowl- tuberculosis is a distinct variety. A very small proportion of guinea-pigs, inoculated in the peritoneal cavity with fowl-tubercle, succumb to the disease, though so susceptible to the effects of human or bovine virus. Maffucci maintains that guinea-pigs have an immunity, and that rabbits rarely develop a generalised tuberculosis. Cultures are not identical in appearance with those obtained from man, and on microscopical examination show many long, thick, and branched forms, which are only rarely found in cultures from a human source. Stamping-out System. — In 1888 a Departmental Committee was appointed to inquire into pleuro-pneumonia and tuberculosis, and they considered that legislation ought to be directed not only to the protection of cattle from tuberculosis, but also to prevent the possibility of the disease being communicated to man. The following extracts are from the recommendations of the Committee, which were made on the lines of : — A. PREVENTION. B. EXTIRPATION. A. — Preventive Measures. These should include provision for : — Improved hygiene of cattle sheds, etc. (especially in the direction of providing proper ventilation, pure water supply, and adequate disinfection of stalls, etc., wherein tubercular animals have been kept). This has been partly met in the Dairy and Milk Shops Order, but its administra- tion by the local health authorities is at present imperfect ; and we would suggest that it should be much more stringently enforced, and that veterinary inspectors should be given more extended powers of entry into all places where animals are kept. TUBERCULOSIS. 403 Improvement in the hygienic surroundings of animals should include isolation of all suspected cases, precautions against the flesh or milk of diseased animals being given as food to others — e.g., to pigs, fowls, etc. — and care that fodder, litter, and water should not be taken from one animal or stall and given to another. Our attention has been drawn to the frequency with which animals, obviously diseased, sometimes even in the last stage of the malady, are sold in open market. Although in England and Ireland, under the provisions of the Nuisances Removal Act as embodied in the Public Health Act, 1885, the medical officer of health or inspector of nuisances may seize such animals, yet such seizure is rarely performed. We find the veterinary inspector has no power to prevent such sales, or to seize the beasts for slaughter, since tuberculosis is not included in the Contagious Diseases (Animals) Act of 1878. We further find that there is actually a regular trade in such stock infected with tuberculosis, and that they go by the name of " wasters " and " mincers," being frequently slaughtered in the neighbourhood of the larger towns, to which such portions of the meat as are likely to escape the observation of the inspector of nuisances are sent, for the purposes of sale among the poorer inhabitants, and especially for the making of sausages. We are, therefore, very strongly of opinion that power should be given to the veterinary inspector to seize all such animals in fairs, markets, or in transit. Notwithstanding the uniform prevalence of the disease in Europe and elsewhere, there seems to be no reason to apprehend that, with our present regulations for the slaughter of animals at the port of debarka- tion, and for quarantine of those imported for breeding, there is any special danger of increasing the infection in England by introduction from abroad. The danger, however, exists in regard to the stock brought from countries, which are exempt from slaughter on landing, and sub- jected to the ordinary veterinary inspection during the present period of detention of twelve hours. It is, therefore, evident that the present rules for the prevention of the introduction of disease into the United Kingdom from abroad, are incomplete. Since all authorities are agreed that the disease is very marked by heredity, we think it highly desirable that breeders should in their own, as well as in the public interest, discontinue breeding from tuberculous stock. B. — Extirpation. In order to insure the gradual extirpation of tuberculosis, we are of opinion that it should be included in the Contagious Diseases (Animals) Acts for the purposes of certain sections of those Acts, so as to provide : — (a) For the slaughter of diseased animals, when found diseased on the owner's premises. 404 INFECTIVE DISEASES. (6) For the payment of compensation for the slaughter of such animals. (c) For the seizure and slaughter of diseased animals exposed in fairs, markets, etc., and during transit. (d) For the seizure and slaughter of diseased foreign animals at the place of landing in this country. Notification of this disease should not be compulsory, because it may exist without developing any sufficient outward evidence to enable the owner to detect it, and its growth is so slow, that non-notification of its •existence, even in a large number of cases, would do little to nullify the stamping-out effect of the Act of 1878. The powers and responsibilities of inspectors in ordering the slaughter of diseased animals should be the same for tuberculosis as for pleuro- pneumonia, according to section 51 (5) of the Act of 1878. Further, tubercle, though hereditary, is nevertheless much less conta- gious than the other diseases included under the Act of 1878, and it is clear, therefore, that the immediate slaughter of diseased animals would go far to stamp it out, though, doubtless owing to heredity, this stamping- out process would be gradual in its effect. A supplementary report was made by Professor Horsley, in which he expressed the opinion that there ought to be legislation to prevent breeding from diseased animals, and compulsory notifi- cation : — 1. Breeding. Tuberculosis is notorious, even among the laity, as a disease which is transmitted from parent to offspring. This is a fact with which cattle breeders are specially familiar, and which finds strong expression in the evidence attached to this report. Further, this generally received truth has been completely confirmed by the results of scientific investigation, as is also duly set f6rth in the report. Considering, therefore, the extreme importance of this point, I think that the act of wittingly breed- ing from animals so affected should be made an indictable offence. The only objection that can be raised to such legislation, which if effected would prevent the dissemination of the disease among cattle in this country, is that, owing to the present state of want of knowledge among cattle owners, and even veterinary surgeons, of the early symptoms, and physical signs on examination, of this disease, prosecutions would occa- sionally occur in cases in which no fault could properly be attributed to the owner, and that, therefore, such prosecutions would be needlessly vexatious. Considering, however, the extreme rarity with which such cases would occur, and that, as in the matter of non-notification, each case would be tried before district magistrates on its own merits, this objection is •deprived of the force it might have possessed. TUBERCULOSIS. 405 2. Notification of the Existence of the Disease. This point' requires no explanation, since it is clear that, unless the veterinary inspectors or authorities receive information of occurrence of diseases, it is impossible to ensure the thorough carrying out of the provisions of the Contagious Diseases (Animals) Act. That deliberate non-notification should be punished cannot be doubted by any one. Objection, however, to legislation in this direction has been put forward, on the same grounds as those upon which the prevention of breeding from diseased animals was contested. As, however, I consider that these objections have been already shown to have no weight, I recommend that both the forbiddance of breeding from diseased animals, and the notification of the disease, should be included in any legislation for tuberculosis. The difficulty referred to by the Committee, is presented by cases of the disease which cannot be detected by the ordinary methods of examination, and might possibly be overcome by the use of tuberculin as a diagnostic agent. CHAPTER XXIX. LEPROSY. — SYPHILIS. — RHINOSCLEROMA. — TRACHOMA. LEPROSY. LEPROSY occurs in three forms — tubercular, anaesthetic, and mixed tubercular. It may be classed with the granulomata, as the most common form of the disease is characterised by deposits in the skin, mucous membrane, and internal organs. These deposits are composed of small cells, and large cells resembling giant cells. The cells become deposited in the surrounding tissues, and so the tubercle enlarges, involving the epidermis and developing into an ulcerating sore ; or, after a certain stage of development, beginning to decline, and finally leaving a puffy discoloration. In the ansesthetic form the cells invade the connective tissue of nerves. In the mixed form the varieties occur together, but the tubercular character predominates. Tubercular leprosy commences with the development of an erytheinatous patch, which becomes infiltrated, and finally tubercu- lated, the tubercles varying in size from a millet seed to a marble, or even larger. The eruption on the head and face produces a characteristic leonine expression. The progress of the disease is very slow. After death the following changes may be found in the internal organs : Cirrhosis of the liver and spleen, enlargement of the lymphatic glands, and a condition of the lungs corresponding to cheesy bronchial pneumonia. In the ansesthetic form patches develop on the skin, which become anaesthetic ; ulceration follows, and the fingers and toes, or the entire hand and foot, may slough off. The disease is undoubtedly communicable, but the infectivity is of a very low type. The infectiousness is illustrated by the well-known case of Father Damien. Arning inoculated a man named Keanu, a condemned criminal, and leprosy developed three years afterwards, but this case 406 LEPROSY. 407 is not regarded as conclusive, as the man had a family history of leprosy. The disease has never been known to spread from patients in this country, who have contracted the disease abroad. The bacilli of leprosy were first observed by Hansen in 1874, and subsequently fully described by him, and his observations confirmed by Neisser, in 1879. Bacillus Leprae. — Rods 5 to 6 /x in length and 1 /x in breadth. The bacilli are straight or curved, resembling very closely the tubercle bacilli. They are present in the leprous tubercles of the skin and mucous membrane, in the lymphatic glands, and in the liver, testicles, and kidney; and in the nerves in the anaesthetic variety. They are found between the cells, and in colonies in the cells. They stain readily with the aniline dyes, especially by the Ziehl-Neelsen and Gram's methods. The bacilli are found in extra- ordinary numbers in the skin; and they are rather straighter than tubercle bacilli, and stain more readily. Numerous unsuccessful attempts to cultivate the bacillus have been made by many bacteriologists. The author has made repeated inoculations upon glycerine-agar, upon which the tubercle bacillus grew abundantly, but always with disappointing results. On the other hand, Bordoni-Uffreduzzi showed the author a cultivation which he had obtained from the bone marrow of a leper. The cultivation was made on blood serum and glycerine, and cover- glass preparations resisted decolorisation with acid. There were slight morphological differences when compared with the -appear- ance of bacillus leprse in the tissues, and the results were hardly conclusive. The English Leprosy Commission also reported successful cultiva- tion of the leprosy bacillus. The author had the opportunity of examining one of the first cultures received in this country, and found that the bacilli stained deeply in ordinary cover-glass pre- parations, they did not resist decolorisation by the Ziehl-Neelsen method, and they corresponded in culture with one of the varieties of Bacillus subtilis, commonly found on the skin. Inoculation of animals has given equally unsatisfactory results. Numerous experiments have been made by Beaven Rake on small animals and birds, with invariably negative results. The blood of leprous patients, tubercles from the living subject, fragments of the skin and of the internal organs after death, have been inoculated by different observers without result. Melcher and Ortmann alone claim to have produced really definite results. These observers excised leprous tubercles from the living subject, a'nd inoculated fragments 408 INFECTIVE DISEASES. in the anterior chamber of the eye of rabbits. The animals died after some months with extensive deposits in the ccecum, lymphatic glands, spleen, and lungs. These tubercles varied in size from a pin's head to a millet seed, and contained bacilli, resembling leprosy bacilli in their staining reactions. The question naturally arises whether the lesions were really indicative of leprosy or tuberculosis. Until the experi- ments are independently confirmed, and the result of inoculation differentiated from tuberculosis, it would be rash to accept these experiments as conclusive. It has been suggested that tuberculosis and leprosy are identical. There is a similarity in the bacilli and in the lesions of leprosy and tuberculosis, the injection of tuberculin produces a reaction in leprosy nodules, and many lepers die from tubercular disease of the lung. But while tuberculosis is very readily transmitted to guinea-pigs and rabbits by inoculation of fragments of tubercular tissue, leprosy is inoculable, if at all, in most exceptional instances. The bacilli of tubercle are cultivated with the greatest facility, the bacilli of leprosy, if at all, only with exceptional difficulty; tubercle bacilli are found in giant cells, leprosy bacilli in the so-called leprosy cells. Leprosy bacilli are straighter than human tubercle bacillir and differ slightly in their behaviour to staining reagents. On the other hand, the morphological differences are not greater than those existing between different forms of tubercle bacilli obtained from tuberculosis in animals and birds. It would be premature to regard leprosy as a variety of tubercle until cultivations of the bacillus have been obtained, and carefully compared with those of the tubercle bacillus. Differences in morphological details and results of inoculation would then carry less weight as a means of differentiation. The tubercular pneumonia of lepers would be regarded, if the bacilli are identical, as a development of leprosy in the lungs, and not, as at present, a result of double infection with tuberculosis. METHODS OF EXAMINING THE BACILLUS OF LEPROSY. Cover-glass preparations may be made in the ordinary way, or by a special method, which consists in clamping a nodule with a pile clarnp until a state of anaemia of the tissue is produced. On pricking with a needle or sharp knife a drop of clear liquid exudes, from which cover- glass preparations may be made, and stained by Neelsen's method. For sections the author prefers Neelsen's method and methylene-blue. They can also be stained by Gram's method, which, as a rule, brings out very clearly the beaded appearance of the bacilli. DESCRIPTION OF PLATE XIY. Bacillus Leprae. FIG. 1. — From a section of the skin of a leper. The section is, almost in its entirety, stained red, and, with moderate amplification, has a finely granular appearance. Stained by the Ziehl-Neelsen method (carbolised fuchsine and methylene-blue). x 200. FIG. 2. — Part of the same preparation with high amplification, showing that the appearances described above are due entirely to an invasion of the tissue by the bacilli of leprosy, x 1500. ; - , : -.- J Plgl. Tig 2 BACILLUS LEPTLE ^SrtAtfp OF THB UNIVERSITY Of LEPROSY. 409 Method of Babes. — Preparations are stained in rosaniline hydrochlorate in aniline water, decolorised in 33 per cent, hydrochloric acid, and after- stained with methylene-blue. Stamping-out System. — The history of leprosy in the British Islands during the Middle Ages, and the conditions under which it both increased and declined, have been discussed by several writers. A large number of institutions of a charitable and ecclesiastical character were established in endemic areas and were occupied by the lepers either voluntarily, or compulsorily by means of the Act De leproso amovendo. These institutions were to a very small extent a means of segregation. According to Dr. Newman the disease, which had reached its zenith about the twelfth or thirteenth century, began to decline from that time owing to " a general and extensive social improvement in the life of the people, to a complete change in the poor and insufficient diet (which it is evident consisted far too largely of bad meat, salt, putrid and dried fish, and an almost entire lack of vegetables) and to agricultural advancement, improved sanitation and land drainage." Of all the unfavourable conditions it would appear that food in some way was especially associated with the cause of the disease, either by introducing the bacillus or by rendering the tissues a suitable soil for its reception and development. In other countries segregation has been attempted voluntarily or compulsorily, but it has never been completely carried out. There can be very little doubt that the presence of a leper in a healthy community is no greater source of danger than the presence of an individual suffering from tuberculosis, but, for other reasons, voluntary isolation should be carried out as completely as local circumstances will permit. The Leprosy Commission in India recommended — (a) That the sale of articles of food and drink by lepers should be prohibited, and that lepers should be prevented from following certain specified occupations. (b) That the concentration of lepers in towns should be discouraged. (c) That Leper Asylums should be established in which lepers might live voluntarily. (d) That Leper Farms scattered over the country should be encouraged. (e) That the few children who are born of lepers should be removed to Orphanages. They concluded that by means of improved sanitation and good dietetic conditions a diminution of leprosy will result. 410 INFECTIVE DISEASES. © ® %1 SYPHILIS. Syphilis is a disease peculiar to man, and communicable 'only by inoculation. The local infection is followed by a period of latency, and by a period during which generalised eruptions appear. One attack confers immunity from future attacks. The virus in its most virulent form is found in the primary seat of inoculation, and in the indurated glands which follow. It is also supposed to be present in the blood and secretions. Lustgarten, Eve and Lingard have found bacteria which they believed to be specific. Bacillus in Syphilis (Lustgar- ten).— Rods resembling the bacilli of leprosy and tuberculosis, 3 to 4 ^ long, -8 //, thick. Two or more colourless, ovoid points in the course of the rod are visible with a high power ; it is thought that possibly they are spores. The bacilli are always found in the interior of nucleated cells, which are more than double the size of leucocytes. They have been ob- . „. ..^w.,** . served in the discharge of the fc'fij^ primary lesion, and in tertiary «V-&.^: gummata. f ?$*$/'• Alvarez and Tavel state that an identical bacillus is found in '^l^v;; normal secretions (smegma). Eve and Lingard have described a bacil- lus associated with specific lesions, which differs from the above in its morphology and behaviour towards FIG. 176.— WANDERING CELL CON- staining reagents. TAINING BACILLI ^ rim* ^a&'viWdsssww*'*' FIG. 175. — COVER-GLASS PREPARA- TION OF PUS FROM A CHANCRE, X 1050 (LUSTGARTEN). •: #&, METHODS OF STAINING THE BACILLUS OF SYPHILIS. Method of Lustgarten : — Sections are placed for twelve to twenty-four hours in the following solution, at the ordinary temperature of the room, and finally the solution is warmed for two hours at 60° C. : — Concentrated alcoholic solution of gentian-violet . 11 Aniline water . ..... 100 SYPHILIS. 411 The sections are then placed for a few minutes in absolute alcohol, and from this transferred to a 1-5 per cent, solution of permanganate of potash. After ten minutes they are immersed for a moment in a pure concentrated solution of sulphurous acid. If the section is not completely decolorised, immersion in the alcohol and in the acid bath must be repeated three or four times. The sections are finally dehydrated with absolute alcohol, cleared with clove-oil, and mounted in Canada balsam. By this method the bacillus is distinguished from many bacteria, but not from the bacilli of tubercle and leprosy which are stained by this process. Method of De Giacomi : — Cover-glass preparations are stained with hot solution of fuchsine containing a few drops of perchloride of iron. They are then decolorised in strong perchloride of iron, and after-stained with vesuvin or Bismarck- brown. Method of Doutrelepont and Schiltz : — Sections are stained in a weak aqueous solution of gentian-violet and after-stained with safraniu. The nature of the contagium in syphilis is unknown. Protective Inoculation. — Inoculation of the virus, or syphilisa- tion, as a protective measure, was at one time practised and strongly advocated ; but it is rightly regarded in this country as dangerous and unjustifiable. From the experiments of Bicord it would appear that the local results in the vesicular stage resemble the results of the inoculation of virulent vaccinogenic grease or horse- pox. The inoculation goes through the stages of papule, vesicle, ulcer, scab, and scar. The accidental inoculation which occurs in cases of vaccino-syphilis may so closely resemble the results of inoculation with very virulent cow-pox, that it is sometimes difficult to decide as to the exact nature of these cases. RHIXOSCLEROMA. Rhinoscleroina is a rare disease, resembling lupus, and pro- ducing in the nostrils and neighbouring parts nodular swellings, composed of granulation-tissue. The disease is met with in America, Egypt, Austria, and Italy. There are no giant cells, but peculiar large cells, which were first described by Mikulicz. Frisch discovered bacteria in sections, and Cornil and Alvarez pointed out the existence of a capsule. In morphology and cultivation they resemble, according to Dittrich, Friedlander's pneumococcus. They are probably identical with this micro-organism, and Paltauf and Eiselsberg, and others, found that they produced septicaemia in rabbits and guinea-pigs. Bacterium of Rhinoscleroma (Bcudttus of Rhinosderama, 412 INFECTIVE DISEASES. Cornil and Alvarez). — Cocci and short rods, 1*5 to 3 /x, in length,. *5 to '8 JJL thick. Deeply coloured points or granules may occur in the course of the rods when stained, but it is very doubtful whether these can be considered as spores. The bacteria are en- capsuled, the capsule being round when enclosing a coccus, and ovoid when enclosing a rod. The capsule is composed of a tough resisting substance; two or more capsules may unite by fusion, enclosing two, three, or a greater number of rods. The bacilli were observed in sections of the tumours, which developed on the lips and in the nasal and pharyngo-laryngeal regions. METHOD OF STAINING THE BACILLUS OF RHINO- SCLEROMA. Method of Cornil and Alvarez : — Sections are immersed in a solution of methyl-violet (B) for twenty- four to forty-eight hours, with or without the addition of aniline-water ; and are then decolorised after treatment with the solution of. iodine in iodide of potassium. If the sections are left to decolorise in alcohol for forty-eight hours, the capsule is rendered visible. TRACHOMA. Trachoma is a disease of the conjunctiva, common in Egypt. The new growth is composed of round cells, and may be regarded, according to Kartulis, as the chronic stage of either gonorrhoeal or Egyptian ophthalmia. Koch failed to find any micro- organisms in the swollen lymph follicles. Sattler asserted that he had culti- vated a micrococcus which produced the disease when inoculated on the conjunctiva. Other observers have found the common pyogenic micrococci in the secretions, especially Staphylococcus pyogenes aureus and albus. CHAPTER XXX. ACTINOMYCOSIS. — MADURA DISEASE. ACTINOMYCOSIS. ACTINOMYCOSIS belongs to the class of infective granulomata. It is a chronic inflammatory affection characterised by the presence of a special microphyte, which by irritation produces a neoplasm, composed -of round cells, epithelioid cells, giant cells, and fibrous tissue. These neoplasms form nodular tumours of various sizes. In some cases there is a tendency to develop very large tumours, and in others to break down early and suppurate. In cattle, cretification takes place in the fungus tufts. Actinomycosis closely resembles tuber- culosis in its histological characters. The disease attacks man, horses, cattle, and pigs. Many interesting observations have been made upon the origin of this disease in man. Two cases have been recorded in support of the theory of direct infection from the cow. Stelzner described a case of actinomycosis in a man who had had the care of animals, some of which had suppurating glands. Hacker had a case of actinomycosis of the tongue in a man who had charge of cows, one of which had a tumour of the jaw which he had opened. On the other hand, Moosbrugger found that out of 75 cases, 54 were in men, and 21 in women, including 2 children. In 11 of these men the occupation was not stated. In 33 their occupation did not bring them into contact with diseased animals ; they were, for example, millers, glaziers, tailors, shop people, and students. Only 10 cases occurred among farmers, peasants, and farm-labourers, and in only one case out of the 10, had the patient been brought into contact with diseased animals. Out of the 21 women, there were only 4 peasants, and none of them had been associated with diseased cattle. Infection by the flesh of diseased animals has also been dis- cussed. But there is no evidence of prevalence of the disease 413 414 INFECTIVE DISEASES. among slaughterers and butchers, who would be particularly liable to it, if flesh were a source of infection. The chances of infection by ingestion are minimised by the flesh being almost always cooked. Actinomycosis occurs also in pigs, and pork is very often eaten in an uncooked state ; but Israel has pointed out that this may probably be excluded, as many of the cases occurred among strict Jews. The evidence points to the disease originating in man and lower animals from the same source, and there is a very strong suspicion attached to cereals. This view is supported by important obser- vations, with reference to the part played by cereals in inducing the disease in cattle, and it gains additional support from a case described by Soltmann, where the disease resulted from an awn of wall barley. A boy, aged eleven, accidentally swallowed an awn of Hordeum murinum. He became very ill, and suffered great pain behind the sternum, extending to the back. An abscess formed, covering an area extending over six intercostal spaces, and when opened, the awn of this grass was found in the evacuated pus. The pain, however, continued, and fresh deposits occurred, and when the boy was taken to the hospital, the ray-fungus was detected. Possibly the spores of the fungus can be conveyed both by air and water. This disease in cattle has long been known in this country, but its various manifestations were either mistaken for other diseases, or simply received popular names. Indeed, the various forms are still familiar to many as wens, clyers or crewels, scrofulous, tuber- cular or strumous abscesses, polypus, lymphoma, cancer of the tongue, scirrhous tongue, indurated tongue, ulcerated tongue, cancer of bone, bone tubercle, osteo-sarcoma, fibroplastic degeneration of bone, spina ventosa, and carcinoma. Bovine antinomycosis is especially prevalent in river valleys, marshes, and on land reclaimed from the sea. The disease occurs at all times of the year, but general experience leads to the belief that it occurs more commonly in the winter. It is more frequently met with in young animals, and usually occurs between one and three years, but it may be found at almost any age, and probably affects equally both sexes. There is little if any evidence to show that the disease is heredi- tary. In numerous cases, the family history has been most carefully inquired into by the author ; and in the case of some imported pedigree animals, the disease was quite unknown on the farm where they had been bred. The tongue is so commonly the seat of the disease, that suspicion ACTINOM YCOSIS. 415 at once falls on food as the means by which the parasite is conveyed. Skin wounds produced by rubbing against the mangers, posts, or wire fencing, may also become infected. The evidence is very strong in favour of believing that the micro- organism gains access to the system through wounds or lacerations of the mucous membrane and skin, or through carious teeth. It has been pointed out that the common occurrence of the disease at the time of the second dentition may be owing to the wounds produced in the alveolar mucous membrane by the shedding of the teeth. Experience also points to straw being sometimes a factor in the production of the disease, and it is possible that thistles and frozen roots also, by wounding the mucous membrane, may afford a way for the entrance of the micro-organism. The disease in the jaws, both in man and in cattle, is very commonly associated with carious teeth. The cowsheds, pastures, and drinking tanks may become infected with the discharges from diseased animals. The discharge con- taminates the fodder in the sheds, and falls on thistles and siliceous grasses in the pasture, which may first wound, and then introduce t]je micro-organism. The discharge is also coughed out of the mouth, and expelled from the nose, in cases in which a tumour in the pharynx, or the nasal chambers, has undergone suppuration. Jensen believed that the disease was produced by different kinds of grain, especially when cultivated on ground reclaimed from the sea. He mentions an instance of a farm, where nearly the whole of the young stock, about thirty in number, had actinomycosis after feeding on mixed forage, grown on a certain field. Two years after- wards the same disease occurred in the same stalls in four animals, after being fed on barley-straw from the same field. According to Jensen, the fungus grows on grain, husks, and straw of different cereals, but most abundantly on barley, which is also the most likely to wound the mucous membrane. Johne's observations tend to corroborate this view, for in twenty-two out of twenty-four cases in which he found barley sticking in the tonsils of pigs, he found the beard thickly beset with a fungus very similar to, if not identical with, the ray-fungus. These observations are of great interest in connection with Soltmann's case. Experience points to the belief that the disease is not readily communicable from animal to animal, and it is possible that when it affects a large number of cattle in a herd, the same causes have been acting to produce the disease in a number, which in another in.-tunce may only produce it in one. At the same time, isolated 416 INFECTIVE DISEASES. cases -are possibly not quite so common as they are reported to be. It is well known that, as a rule, the services of a veterinary surgeon are not called for except in hopeless or very severe cases. The cowmen themselves, in many districts, treat the cows successfully, and then send them into the market, and thus the existence of previous cases may not have come to the knowledge of the veterinary surgeon. Historical. — In 1845 Professor von Langenbeck, of Kiel, made notes of a case of vertebral caries in a man, and prepared drawings of peculiar bodies in the pus from an abscess. The drawings were published together with a reference to the case by Israel in 1878. There can be little doubt that these structures were the fungi of actinomycosis. But the first to publish observations was Lebert in 1848. Lebert received from M. Louis some pus, of a thick, almost gelatinous consistency, which had been obtained from an abscess of the thoracic wall in a man aged fifty. The patient had been attacked four months previously by a pulmonary affection, which was suspected by M. Louis to be cancerous in nature. The pus contained a very considerable number of little spherical bodies of a slightly greenish-yellow colour, about the size of a pin's head. They could be readily crushed between two strips of glass, and on examination with a power of fifty diameters two elements could be distinguished : a soft connective substance, and many hard, narrow, wedge-shaped corpuscles, arranged in a radiating manner. Under a high power these bodies were observed to be -2^ to ^ of an inch in length, -^^ in width at the base, and -.^ in width at the apex. Some of these corpuscles were regular, while others showed one or two constrictions, with intermediate flask-shaped swellings. Lebert tested these structures with reagents, with the following results. The bodies were found to remain unaltered by concentrated mineral acids. Acetic acid freed them from foreign elements adhering to their surface. Solution of caustic potash did not affect them if used cold, but a boiling solution reduced the cuneiform structures to a fine greyish powder without dissolving them. Ether, alcohol, and chloroform had no effect upon them when used either hot or cold. Solution of potash, in which these bodies had been heated, mixed with a solution of sulphate of copper and brought to boiling point, did not offer any uniform red colour, which would have been the case if they had contained albumin. Thus, the chief chemical characters of albuminous and fatty substances were wanting, and they resembled chitine in their behaviour to reagents. ACTINOMYCOSIS. 41 r Lebert bore in mind the possible existence of some helminthic debris, of which these bodies might be booklets, but he sought in vain for echinococci and cysticerci. Actinomycotic pus was later described and figured by Robin. In the illustration accompanying the description, the fungi are most FIG. 177. — SECTION OF LIVER FROM A CASE OF ACTINOMYCOSIS IN MAX. accurately depicted. Robin states that he had found, in two or three cases in the pus of deep-seated chronic abscesses, yellowish grains attaining a diameter of one-tenth of a mm., surrounded by a sort of halo or thin, viscous, finely granular stratum, containing leucocytes. These grains were composed of elements 2 to 6 mm. in length, swollen 27 418 INFECTIVE DISEASES. at one end and tapering off at the other, arranged in a regular series, radiating from a common centre which consisted of granular matter. They were highly refractive, possessed a brilliant centre and sharply -defined outline ; they were dissolved, or at least rendered indistinct, by acetic acid, and proved insoluble in ammonia and ether. The disease in man was next described by Israel in the paper mentioned above. Ponfick was the first to clearly recognise the identity of the disease in man with the disease in cattle, and he described a number of cases in man. Israel subsequently published a work on the subject. The various cases which had been observed up to that date were described, and the disease classified according to the seat of invasion. From this time onwards numbers of cases in man have been described, and various important researches published, of which those of Bostrom and Moosbrugger may be especially mentioned. In England, Acland recognised a case on examining the liver after death (Fig. 177). H. Taylor was the first, in this country, to detect the fungus during the life of a patient. Shattock found .specimens of the disease in museums. Skerrit, Powell and Godlee, Eve, Delepine, Hansome, Poore, Malcolm Morris and others have published cases. In Italy, Perroncito studied the sarcomata of cattle, and claims to have first observed the micro-organism in 1863. In 1875 he described it in the Encyclopaedia Agraria, and, from the negative results obtained by inoculation experiments, was led to regard it, not as the cause, but as a result of the disease. Rivolta of Turin also claims to have been the first to have •discovered the fungus in actinomycosis bovis. As early as 1868 he published a paper on a sarcoma tous tumour of the jaw of -an ox. Hahn of Munich, in 1870, undoubtedly met with the fungus, for he states that in a case of " wooden-tongue " he found characteristic organised structures, which he provisionally described as a species of mould fungus. Bellinger was the first to recognise the nature of this disease in cattle. In 1876 he pointed out that new growths occasionally occurred on the upper and lower jaws of cattle, which either started from the alveoli of the back teeth, or from the spongy tissue of the bone, and by increasing in size loosened the teeth. In their progress they destroyed bone, muscles, mucous membrane, and skin. After some time they frequently broke down, forming ulcers, abscesses, and fistulas ; but in some cases tumours were formed, which attained ACTINOMYCOSIS. 419 the size of a child's head. Bellinger stated that this disease had been known by various names, — Osteosarkome, Winddorn (Spina ventosa), Knochenkrebs, Knochenwurm ; in other instances it had been regarded as bone tuberculosis, or mistaken for a simple chronic glossitis. Among breeders of cattle and owners of stock in Germany it had been known under the following names : Ladendruck, Laden- geschwulst, dicker Backen, £dckel, Kinnbeule, Kiefergeschwulst, etc. Bollinger pointed out that these swellings consisted of several centres of growth, bound together by connective tissue. They were often as large as a walnut or a hen's egg, and of a pale yellow colour and moist appearance. The cut surface presented yellowish- white, suppurative foci, while in other cases the growths had a spongy texture, owing to the formation of lacunae or hollow spaces in a fibrous stroma, which contained a turbid, thick, yellow, caseous pulp. Microscopical examination of the tumour showed that it had a structure like a sarcoma, while the squeezed-out pulp consisted principally of pus cells, granulation cells, fat granules, and granular detritus. In addition, there were numerous opaque, pale-yellow, and coarsely granular bodies of different sizes, which had a mulberry- like appearance, and were sometimes encrusted with chalk. After careful examination Bollinger found that these bodies were true fungi, and he further maintained, from the constancy of their appearance in all parts of the sarcomatous growth, that they were not accidental, but of pathogenic significance. This was found to be the case, not only in fresh preparations, but in old specimens preserved in the museum. This remarkable form of mycosis was observed by Bollinger, not only in the upper and lower jaws, but also in the tongue. It had long been observed that the tongue was sometimes covered with more or less tubercular growths, scattered abundantly over the surface of the mucous membrane, mostly the size of a millet seed or hemp seed, but often reaching the size of a cherry or walnut, or even larger. In the fresh state these nodules were greyish -white, and semi- transparent, but they soon became cloudy or distinctly puriform in the centre ; they were surrounded externally with a connective tissue capsule. If the nodules were situated on the surface of the tongue, destruction of the mucous membrane very readily followed, leading to the formation of ulcers. The tongue also might become affected with an interstitial glossitis, which often, in spite of the partial atrophy of the muscular fibres, led to a great enlargement and wood-like hardness of the tongue. On account of this peculiar character, such a tongue was long known in South 420 INFECTIVE DISEASES. Germany as Holzzunge. In other cases the condition was regarded as " tubercle of the tongue," " chronic sarcoma," " chronic interstitial glossitis," or simply " degeneration of the tongue." Bollinger described this disease as occurring in cattle of all agesy developing itself gradually, and being always incurable. As a ruler the animals were slaughtered, because the diminished mobility and enlargement of the tongue interfered with feeding. He also pointed out that this disease of the tongue was by no means rare, as he had had no less than six such tongues from different parts of Bavaria in the space of a year, and he also had been able to prove the existence of the disease in museum specimens. On further continuing his researches, Bollinger found the same fungus in tumours which occurred in the pharynx, larynx, and the mucous membrane of the stomach. These tumours were very common in the throat in some parts of North Germany, where as many as 5 per cent, of the animals had been known to be affected. The disease frequently occurred in the form of subcutaneous neoplasms, called Lymphome, Hohzgeschivulste, Fibrome, Tuberkel, Tuberkel-scropheln. This disease also appeared in the form of abscesses, which were called, in many districts, Schlundbeulen. These growths were found in the neighbourhood of the parotid gland, the larynx, and pharynx, and were similar in every respect to the affection of the jaw. They were described as starting apparently from lymphatic vessels in these parts. Bollinger discovered the fungus in a case of so-called fibroid of the second stomach of a cow, a spongy growth nearly the size of the fist ; and he believed that in another case the disease manifested itself in the form of tubercular ulceration of the intestines. Bollinger submitted the fungus to Dr. Harz, a botanist, who described the fungi as mulberry-like masses from *5 to 1 mm. in diameter. They appeared to the naked-eye as opaque, white grains,. and when calcined were difficult to recognise. On slight pressure the tufts of the fungi fell apart into segments of unequal size, each of which appeared to correspond to an individual fungus. The latter was described as beginning at the pointed end of the wedge, with a somewhat cone-shaped basal cell, which, in the absence of a mycelium, perhaps took its place, and bore a great number of short linked hyphse. At the ends of the hyphse there were oval, globular, or elongated club-shaped bodies, the reproductive cells or gonidia. Cultivation experiments, and inoculation of the tongue of a calf with liquid containing the micro-organism, failed. Harz proposed ACTINOMYCOSIS. 421 to call the fungus, from its ray-like appearance, actinomyces ; but what the position of the fungus in nature might be, was difficult to determine. It did not, he believed, belong to the yeast fungi, but to the mould fungi, and might be compared to Botrytis, Monosporium, arid Polyacti.s. Bollinger concluded that there could be no doubt that actino- mycosis occupied an important position in the pathology of cattle diseases. As further evidence of the prevalence of the affection, he remarked that Zippelius of Obernburg had observed in the course of about ten years' practice not less than 254 cases of lymphoma, in the neighbourhood of the larynx and pharynx, besides 157 cases of disease of the jaw ; and Bollinger says that he had very little doubt that the greater part of the former, and very likely all the cases in the jaw, were due to the fungus which he had discovered. In certain parts of Eranconia, according to a communication received from Professor Frank, these tumours of the throat were extremely abundant in cattle. Bellinger's researches were followed by those of Siedamgrotzky, and later by a communication from Johne. Johne described the various forms of the disease which had up to that date been recognised, including a description of actinomycosis of the bones of the jaws, of the fauces, of the larynx, of the oesophagus, of the stomach and intestinal canal, and of the udder. He carried out a series of experiments, by which it was clearly established that the disease could be communicated from cattle to cattle. Previously Bollinger, Harz, Perroncito, Ponfick, Siedamgrotzky, and Johne had failed, but subsequently by employing fresh material from the living animal, both Johne and Ponfick succeeded. Siedamgrotzky not only confirmed Bellinger's researches, but he described the presence of the fungus in so-called " multiple sarcomas '' of the mucous membrane of the O3sophagus. Rabe described the presence of the fungus in tumours known as Winddorn, and pointed out that, in at least one case, he considered that the disease had been carried by the lymphatics. There were eleven subcutaneous tumours in a row on the face, which were connected by swollen, rope-like, lymphatic vessels. They appeared to be secondary to a growth on the nostril, the size of a hen's egg. Perroncito described a case of " sarcoma " of the intestines and stomach, which proved to be actinomycosis. Many additional communications were made on the subject of this disease. Ponfick produced it in the lungs by intravenous injection, and subsequently three cases occurring naturally in the 422 INFECTIVE DISEASES. practice of veterinary surgeons were published. They not only deserve especial mention, but as this form of the disease appears to be so seldom recognised, they will be given in detail. Plug described a case in the lungs. The cow had been out of health for four weeks, did not eat, and had a cough, and two days previous to the visit had become rapidly worse. Schmidt found dyspnoea with abdominal respiration ; the nostrils were dilated, the head protruded, and the mouth kept open. There was dulness on percussion, and crepitation. The animal was killed, and the lungs, which alone were diseased, were sent to Plug. The pleura on exami- nation was normal, but beneath it were numbers of miliary tubercles, many equal in size to a pin's head. On section the lung had a granular appearance from the presence of countless numbers of minute deposits, which all had the appearance of grey tubercles ; in none was there any central softening. They were present in enormous numbers around the bronchi, and in the vessels of the interlobular tissue. Microscopical examination showed, in the middle of most of these nodules, the presence of greenish-yellow, radiating bodies, which under a high power appeared to be undoubtedly actinomycotic granules. In many there were only rudimentary fungi consisting of four or five clubs ; there was only one rosette in each tubercle. The fungus was surrounded by round cells and fibrous tissue. Larger nodules resulted from the agglomeration of several tubercles, or from diffuse infiltration of round cells in the neighbour- hood of a tubercle. Hirik met with a somewhat similar case. A ten-j^ear-old cow was slaughtered, and in the middle lobe of the right lung there were yellowish nodules about the size of a pea, scattered over an area the size of the palm of the hand. These nodules were not at first sight distinguishable from ordinary tubercles, but on closer inspection they appeared to be somewhat different, and could be easily shelled out from the thickened lung tissue. On making a section, pus welled up at several points, and contained yellowish, calcareous particles. These particles, on microscopical examination, were found to be strongly calcified tufts of the actinomyces embedded in granulation cells. Addition of hydrochloric acid dissolved the calcareous matter, but had no action on the fungus. Pusch described a third case. The lungs of a cow, which had been killed on suspicion of having pleuro-pneumonia, were sent for examination. The front lobe of the left lung was collapsed and firm, the pleura was thickened and opaque ; the larger bronchi were enlarged, filled with pus, and their walls thickened. In the posterior ACTINOMYCOSIS. 423 lobe of the left lung there was a cavity the size of the fist, which had been opened, and the contents had, for the most part, escaped mf what remained was a greyish, purulent liquid, full of yellowish bodies. By the side of this cavity there was another collection of pus, the size of a walnut. In the lower part of the second lobe of the right lung there was a firm, grey tumour, the size of a hen'* egg, over which the pleura was much thickened. On section thi& was cavernous, with similar purulent contents, and yellow grains. These grains under the microscope proved to be ray-fungi. The wall of the cavity consisted of dense connective tissue lined with a soft granulation tissue, bathed in pus. There was no disease of any other parts in 'this casa, so that it corresponded in this respect with the two previous ones. Pusch adds that it was difficult to determine whether the organism had gained access to the lungs by the blood- vessels, or by the inspired air. In his case he inclined to the latter view, and concludes by saying that the organism is probably very common and attached to the most varied objects, from which it may be conveyed by the air. Pusch refers in the same paper to an interesting case which occurred in the practice of Eggeling. The latter had under his care a cow with extensive paralysis. The spinal cord was compressed by a compact swelling in the neck, consisting of the nodules of actinomycosis. There were no manifestations of disease in any other part of the body. Prevalence of the Disease. — The author found that the disease was not generally recognised as a common affection of cattle in this country, in spite of the interest excited by the work of Fleming, to wiiom is due the credit of first recognising a case in England. In 1887 there was a disease prevailing in Norfolk, and in the following year outbreaks were investigated by the author in Essex, Hertfordshire, Cambridgeshire, and Middlesex. In the Norfolk outbreak the author found on one farm 8 per cent, of the beasts affected with the so-called " wens " or " sitfasts," which proved on microscopical examination to be cases of actinomycosis. These growths had previously been described in veterinary text-books as the result of strumous or scrofulous inflammation ; but in all the specimens of wens received from this country and the colonies, the author has been able to demonstrate the presence of the ray- fungus. A case of pulmonary actinomycosis, with grape-like growths on the pleura, indicated that wens were not the only manifestation of this disease, which had teen lost sight of under the designation of 424 INFECTIVE DISEASES. tuberculosis. Many other cases were examined, and the disease was shown to be prevalent in this country. FIG. 178. — From a photograph of a Norfolk steer. There is a growth about the size of an orange in front of the throat, an example of a so-called " scrofulous " or "strumous" tumour. This growth was associated with a large polypoid growth in the pharynx which, by interference with deglutition, produced emaciation (Fig. 180). In Australia actinomycosis commonly occurs in the form of tumours of the upper and lower jaw, which were attributed to " cancer " or to " scrofulous inflammation." The disease is still commonly known in Australia as " cancer " and " lumpy jaw." Reports of the prevalence of actinomycosis in the United States have been published, by the Board of Live Stock Commissioners for the State of Illinois. In their Report for 1890 several interesting communications were pub- lished. Mr. Casewell, State Veterinarian, investigated an outbreak of this disease, known also in America as " lumpy jaw,' FIG. 179.— A NORFOLK HEIFER WITH A LARGE ' WEN " IN THE PAROTID REGION. ACTINOMYCOSIS. 425 -on a farm in Yates City, where there were 80 head of cattle, and 1C were found to be suffering from actinomycosis. Mr. Casewell FIG. 180.— Photograph of a steer nearly three years old, but about the size of a yearling. The emaciation and deplorable aspect recall the appearance of " a piner "or " waster " (tuberculosis). reported that the disease was prevalent in nearly every county in that State, and that in his opinion it was spreading. In one instance 109 cases were slaughtered. Actinomycosis in Relation to Tuberculosis. — When we consider the very high percentage of cases of tuberculosis which has been reported in some localities, the im- portance of differentiating actino- mycosis from tuberculosis cannot be over-estimated. The very great contrast in. the appearance of the micro-organisms in the two cases renders this a very easy matter for the pathologist. But practical veterinarians and breeders of cattle are liable to mistake some . manifestations of actinomycosis for tuberculosis. T, . - ,, FIG. 181.— ACTIXOMYCOTIC NODULES It is of the greatest im- FROM THE PLEURA. portance to bear in mind that wens or clyers are really not tubercular, but actiiiomycotic ; and 426 INFECTIVE DISEASES. that a condition of the lungs may occur as the result of actino- mycosis, which from the naked-eye appearances may be mistaken for "grapes" or " angleberries." It will be well also to remember in connection with the above remarks, that extreme emaciation may result in actinomycosis, producing a condition which, without a post-mortem examination, would probably be attributed to tuber- culosis, the animal being regarded as a " piner " or " waster." If these possible fallacies are taken into account, the excessive per- centage of tubercular cases so commonly reported will be very considerably reduced. There is no evidence to show that the flesh of animals suffering from actinomycotic tumours is unfit for human consumption. In very severe cases it is unwholesome, but there is no evidence that it can produce actinomycosis in man. MANIFESTATIONS OF ACTINOMYCOSIS IN MAN. (I.) Invasion by the Mouth and Pharynx. — The fungus may gain access through carious teeth, or wounds or fistulse of the jaw, and very possibly by inflammatory processes in the pharynx and tonsils. The disease attacks the 'lower jaw most frequently. The turnout- is found in close connection with the bone, or in the sub-maxillary or sub-mental regions, and also in the prse-tracheal region. It occurs, though rarely, in the interior of the bone. In a case described by Israel, which occurred in a woman aged forty- six, there was a small tumour about the size of a cherry attached to the external surface of the lower jaw, with an opening through which a probe could be passed into the bone. The tumour was incised and scraped away, and a cavity discovered in the bone, admitting a small sharp-spoon. Later, a further operation was performed : the periosteum was detached, the cavity of the bone enlarged, and the contents scraped out, consisting of granulation tissue, fragments of bone, and the yellowish fungi. At the bottom of the cavity the fang of the canine tooth was found. ISTo return of the growth occurred. The first cases of actinomycosis which were observed in America were connected with the jaw. In 1884 Dr. Murphy described two cases at Chicago. The first was that of a woman aged twenty-eight. Two weeks previously she had suffered from severe toothache, with swelling in the throat and great pain in swallowing. It disappeared after poulticing, but she was again attacked with toothache, and a swelling appeared on the angle of the jaw on the left side. ACTINOMYCOSIS. 427 mouth could not be opened without difficulty ; the tonsil was much enlarged, and pus was set free on incision. She still suffered with toothache, and a small swelling now occurred on the left side of the neck below the jaw. She had several carious teeth. The swelling, which was about the size of a walnut, was punctured, and a drainage tube inserted ; a creamy-looking discharge containing yellow granules continued to escape, but the swelling and induration increased. A further operation was decided upon. The carious tooth was removed, and a probe passed into the alveolus showed a communication with the external wound ; the angle of the jaw was chiselled aw~ay, and the alveolus scraped out. lodoformed gauze was applied, and the case recovered. The second case was a man aged eighteen, who had also suffered with severe toothache and swelling at the angle of the jaw. On examination a carious tooth was noticed. The swelling was well marked, and there was fluctuation ; it was as large as a pigeon's egg, and situated below the jaw. When punctured, thick creamy pus escaped containing the fungi ; the sinus was scraped out, and in ten days the wound was healed. Another swelling appeared, and this was treated as before, and the case recovered. The peculiar feature of these growths is their apparent migra- tion. Israel states that in one case a tumour occurred on the alveolar process, close to carious teeth, and later was close to the edge of the jaw in the sub-maxillary region. From thence it disappeared, and a large swelling formed below the hyoid bone, and after this had been incised and had healed, an abscess formed above the clavicle. Actinomycotic tumours in this region would sometimes appear to correspond very closely with wens or clyers in cattle : they may dis- charge through the skin, and the opening close, or a fistula result ; but they differ, from their tendency to form burrowing abscesses instead of recognisable tumours. In this respect they recall chronic inflammation rather than the sarcoma-like growths in cattle. Cases in which the upper jaw is attacked are not so frequent as those in the lower jaw. The progress is usually described as slow, and there is a tendency for the deep-seated soft parts to be involved, while in the lower jaw there is a tendency for the tumour to come to the surface. There may be burrowing suppuration, or small tumours, which, after a time, fluctuate and form distinct abscesses. These may involve the skin, discharge their contents, and leave fistulous openings. In other cases the disease has been described as extending from the alveolar process to the temporal bone, or the base of the skull, 428 INFECTIVE DISEASES. destroying bones and even reaching the brain ; or the growth may descend by the spinal column, implicating the vertebrae, and travel- ling and pointing in various directions. (II.) Invasion by the Respiratory Tract. — In one recorded case the disease existed for seven years, was localised to the bronchi (Bronchitis actinomycotica), and did not extend into the lungs. The sputum was examined, and contained the characteristic fungus. If the micro-organisms are inhaled they pass into the bronchioles and alveoli, and produce proliferation of round cells, which undergo fatty degeneration. The resulting patches of peri-bronchitis or pneumonia become yellowish-white ; suppuration and haemorrhage from the capillaries follow, and small cavities result, containing pus cells, fat granules, blood, and the fungi. In the neighbourhood of the new growth there is compression of the alveoli, and ultimately the formation of a dense stratum of connective tissue, separated from the cavities by a lining of granulation tissue containing the character- istic fungus. The symptoms are usually obscure ; but the sputum may contain the fungi, which are often visible to the naked eye. The apices of the lungs are not, as a rule, affected. There is con- siderable clinical resemblance to chronic phthisis : cough, night- sweats, pallor, shortness of breath, and haemoptysis are symptoms common to both. Light may be thrown upon the case by the examina- tion of the sputum. The presence of the actinomyces will be positive evidence as to the nature of the disease. The existence of these symptoms, with absence of tubercle bacilli, would lead to the suspicion of actinomycosis, even failing the discovery of the fungus in the sputum. In the second stage the symptoms are more characteristic. The diseas3 spreads to neighbouring parts, and pleurisy commonly super- venes. This extension may involve the peri-pleural tissues. Thus the disease may follow the prse-vertebral tissues, descend behind the insertion of the diaphragm, and point as an ordinary psoas or lumbar abscess ; it may perforate the diaphragm and reach the abdominal cavity. Peritonitis or sub-phrenitic abscess may then result. In some cases adhesions have formed, and the disease has extended to the liver or spleen, or other abdominal organs. The disease may also extend forwards in the direction of the anterior mediastinum and the pericardium. The primary affection of the lung becomes of secondary import- ance. Grave symptoms occur, hectic fever, night-sweats, rigors, and marked pallor. In the third stage, the disease comes to the surface, either over the chest, or in the neighbourhood of the dorsal ACT1NOMYCOSIS. 429 or lumbar vertebrae ; a swelling appears of a livid colour, and if punctured no fluid escapes, but if allowed to make its own way to the surface, the skin gives way, a muco-purulent discharge mixed with pieces of the growth escapes, and the fungi can readily be recognised. (III.) Invasion of the Digestive Tract. — In a case under Chiari, death, with general marasmus, took place at the age of thirty-four, after two years' illness. The mucous membrane of the intestines was almost completely covered with whitish patches, raised in the centre, and covered with yellow and brown granules closely adherent ta the adjacent tissues. The teeth were carious. Small nodules about the size of a pea may be found in the sub- mucous tissue, and in the mucous membrane itself. They soften and form ulcers with undermined edges, the base reaching the muscular layer. They may undergo cicatrisation, but generally the disease extends through the peritoneum to the abdominal cavity, and perforates the bladder or the intestines, or makes its way through the abdominal wall. Symptoms are either absent or not character- istic. The fungus may sometimes be found in the evacuations, or by exploratory puncture. (IV.) Undetermined. — In addition there are a number of recorded cases presenting very varied symptoms and anatomical relations, in which it has not been possible to satisfactorily determine the path of infection. Delepine has described a most interesting case of an actinomycotic tumour of the brain. MANIFESTATIONS OF ACTINOMYCOSIS IN CATTLE. (I.) In the Digestive system we find the disease attacking : — (a) The lips, gums, buccal mucous membrane and palate, and appearing as nodules, wart-like growths, or ulcers. The nodules and ulceration of the palate were well shown in a specimen sent to the author for examination, under suspicion of being the result of severe foot and mouth disease. (b) The upper and lower jaw, where it probably originates in carious teeth, and extending and invading the neighbouring cavities and sinuses destroys the tissues with which it comes in contact, expanding the bones into thin plates or reducing them to the appearance of pumice-stone. (c) The tongue, where we see it most commonly in the form of nodules or wart -like patches under the mucous membrane, with a special tendency to ulcerate, through the irritation of the teeth. These nodules may extend into the deep muscles, and often collect in rows 430 INFECTIVE DISEASES. more or less parallel to the superficial muscular fibres. Complete tranverse sections of the tongue, double-stained, readily show this arrangement, even to the naked eye. Induration of the tongue results from secondary interstitial glossitis. The author has seen, in one case only, a tumour embedded in the substance of the tongue about the size of a small Tangierine orange, and more or less isolated from any surrounding growth. (d) The pharynx, where the disease may \ occur in the form of polypoid growths producing asphyxia. FIG. 182. — A NORFOLK STEER WITH EXTENSIVE ACTINOMYCOTIC ULCER ATION OF THE SKIN OF THE FLANK. (II.) In the Respiratory system we may meet with the disease in : — (a) The nasal cavities, originating primarily there or resulting from extension of a growth from the lips, or the pharynx. (6) The larynx and trachea, generally in the form of polypoid growths, sessile or pedunculated, which arise primarily or occur secondarily, by extension from the tissues in the neighbourhood. (c) The lungs, where the differentiation of the disease is most important, as neoplasms in the lungs, especially in the early stages, and nodular growths on the pleura, may be mistaken for tuberculosis. The disease is very rarely found in connection with the Nervous system (III.), but probably does not so rarely attack the Reproductive system (IV.). ACTINOMYCOSIS. 431 (V.) The skin and subcutaneous tissues are a favourite seat of this disease, producing the so-called wens or clyers so commonly seen in the fen country. A wen is first recognised as a small tumour, the size of a marble or walnut, which increases in size sometimes with great rapidity, and breaks down and discharges its muco-purulent contents through the inflamed and ulcerated skin ; or it may go on increasing, and form a large compact growth, the size of a child's head. These growths when excised, hardened, and cut, have a characteristic honeycombed appearance, produced by the interlacing bands of fibrous tissue, which form a spongy structure, from the interstices of which the fungus tufts and thick yellowish pus have for the most part dropped out. Actinomyces Hominis.— Careful examination of pus from a, case of actinomycosis in man will reveal to the naked eye little yellowish-white or yellow bodies, which a casual observer might mistake for grains of iodoform. On collecting some of the discharge in a test-tube, and holding it between the light and the eye, the tufts of fungi appeared as brownish or greenish-brown grains, embedded in a muco-purulent matrix. On spreading some of the discharge on a glass slip, the largest tufts of the fungus are found to be about the size of a pin's head. They have a distinctly sulphur- yellow colour by reflected light, but appear of a yellowish or greenish- brown tint by transmitted light. With a sewing needle, or a platinum wire flattened at the end into a miniature spatula, the grains can be readily picked out of the discharge, or taken off the dressing, transferred to a clean slide, and gently covered with a cover-glass. Examined with an inch objective, they have the appearance of more or less spheroidal masses of a pale greenish -yellow colour. On removing the preparation from the microscope, and gently pressing down the cover-glass with the finger, the grains flatten out like specks of tallow ; and on again examining with the same power they are found to have fallen apart into a number of irregular and sometimes wedge-shaped fragments of a faintly brown colour, affording a characteristic appearance. By preparing another specimen, and covering it with a cover-glass without completely flattening out the grains, the spherical, oblong and reniform masses of which the tufts are composed can be recognised with a J- in. objective as rosettes of clubs. By examining the peripheral part of a rosette with a Yj-in., and especially after pressing the grains into a thin layer, with or without the addition of a drop of glycerine, the characteristic clubs are most readily demonstrated, and the most varied shapes observed by carefully 432 INFECTIVE DISEASES. examining the form of the individual elements. As in the bovine , fungus, every variation in form is found, from single clubs to clubs with lateral offshoots, clubs bifid at the extremity, palmate or fan-shaped groups, and banana-like bunches. In many cases the clubs are divided by transverse fission into two, three, or more segments. As a rule, the clubs are irregular in shape, and of about equal size, while a few are conspicuous by their length. In other parts of the preparation the clubs are replaced by long slender forms, which are sometimes transversely divided into a number of short links. With suitable illumination many clubs are seen to taper off into slender filaments. In addition there are free filaments, which are twisted, branched, and sometimes distinctly spirilliform. Many of the clubs are, composed of layers differing in their refractive power, and many have the appearance of a cejntrajjchannel. There are also in the preparation small, highly refractive bodies, fat granules, granular detritus, round cells, pus cells, and sometimes blood corpuscles. The grains differ, as a rule, from those from a bovine source, in the absence of that sensation of grittiness so often transmitted to the finger when pressing the cover-glass upon them, and in the slightly greater tendency of the tufts to retain their compact form. By teasing the grains in a drop of water on a slide, and examining the preparation with a -g- or a ~% objective, the explanation of the latter is forthcoming ; for by this process the clubs are gradually washed away, and a central core remains, which is com?- posed entirely of a dense network of filaments. This can readily be observed by using a small diaphragm, and it will be found that the rosettes of clubs are now replaced by tangled masses, having some resemblance to miniature tufts of cotton-wool. These filaments constitute the delicate network which is seen in sections stained by the method of Gram. This can be readily verified by making a cover-glass preparation of the grains, and staining by that method. The characters' of the fungus can readily be studied by proper illumination, without staining. The clubs have a faintly greenish tint, and in form and arrangement are quite characteristic and easily recognisable. Permanent preparations may be made by mounting the fungus in glycerine. DESCRIPTION OF STAINED SPECIMENS. The fungus may be stained in alcoholic solution of eosin in the manner to be described for the bovine organism, or in orarige-rubinr DESCRIPTION OF PLATES XV. AND XVI. Actinomyces. PLATE XV. FIG. 1. — From a preparation of the grains from an actinomycotic abscess in a boy ; examined in glycerine. The drawing has been made of a com- plete rosette examined by focussing successively the central and peripheral portions. Towards the centre the extremities of the clubs are alone visible ; they vary in size, and if pressed upon by the cover-glass give the appearance of an irregular mosaic. Towards the periphery the clubs are seen in profile, and their characteristic form recognised. At one part there are several elongated elements, composed of separate links, x 1200. FIG. 2. — Different forms of clubs from preparations in which the rosettes have been flattened out by gentle pressure on the cover-glass, x 2500. (a) Single club. (#) Bifid club. (0) Club giving rise to four secondary clubs. (cT) Four clubs connected together, recalling the form of a bunch of bananas, (e) Mature club with a lateral bud. (/) Apparently a further development of the condition represented at (e). (^) Club with a lateral bud and transverse segmentation. (Ji) Single club with double tranverse segmenta- tion, (i) Club with oblique segmentation, (j) Collection of four clubs, one with lateral gemmation, another with oblique segmentation. (&) Club with lateral buds on both sides, and cut off square at the extremity. (Z) Club with a daughter club which bears at its extremity two still smaller clubs, (nt) Club divided by transverse segmentation into four distinct elements. (#) Elongated club composed of several distinct elements. (0) and (jy~) Clubs with terminal gemmation. (^) Palmate group of clubs. (r) Trilobed club, (s) Club with apparently a central channel. (£) Filament bearing terminally a highly refractiveoval body. PLATE XVI. FIG. 1. — From a section of a portion of the growth removed from a boy during life. The tissue was hardened in alcohol, and cut in celloidin. The section was stained by Gram's method and with orange-rubin. x 50. FIG. 2. — From the same section. A mass of extremely fine filaments occupies the central part of the rosette. Many of the filaments have a terminal enlargement. The marginal part shows a palisade of clubs stained by the orange-rubin. x 500. FIGS. 3 and 4. — From cover-glass preparations of the fungus teased out of the new growths produced by inoculation of a calf with pus from a boy suffering from pulmonary actinomycosis. Stained by Gram's method and orange-rubin. The threads are stained blue and the clubs crimson (a) In the younger clubs the thread can be traced into the interior of the club (#). In some of the older clubs the central portion takes a yellowish stain, and in others the protoplasm is not continued as a thread, but is collected into a spherical or ovoid or pear-shaped mass. In others, again irregular grains stained blue are scattered throughout the central portion (Fig. 4). x 1200. FIG. 5. From a pure-culture on glycerine-agar. («) branching filaments, (J) a mass of entangled filaments. Gram's method, x 1200. FIG. 6. — From a similar but older cultivation, (a) a filament with spores (J) chains of spores simulating streptococci. Gram's method, x 1200. ACTINOMYCOSIS. 433- and in either case mounted in glycerine. But although the fungus can be detected without any staining process, there may sometimes be doubtful appearances, and then cover-glass preparations should be made and stained by the method of Gram with eosin. The filaments can be readily recognised, and this is of great value, as it forms an additional means for the diagnosis of the disease. In combination with orange-rubin we have a test that is as characteristic and useful as staining for tubercle bacilli. The discharge, scraping from a growth, sputum, or the isolated fungus is squeezed between two- cover-glasses, which are then slid apart ; they are allowed to dry, passed through the flame in the ordinary manner, and then stained. The cover-glasses can be cleared in clove-oil, the excess of clove-oil being removed by gentle pressure between pieces of blotting-paper,, and then the preparation can be mounted in balsam and rendered per- manent. On examination of these specimens the masses of filaments will be found to be stained blue, and the tissue elements pink. These filaments vary very much in extent and character in different pre- parations. In some cases there are masses of short threads, which are either straight, sinuous, or twisted, and branched. In other parts the field is occupied by very short, straight, or curved and sometimes spiral fragments ; in others, again, there are comparatively long strands. On examination with a high power, and with careful illumination, some filaments will be observed to be moniliform, while others are provided with a terminal oval body. There are also free spherical, and oval, bodies stained blue, which represent the spores of the organism. When orange-rubin is used instead of eosin, the clubs will be stained and easily recognised. This method enables one to determine the exact relation of the threads to the- club-shaped bodies ; and this is an interesting point, as it has been suggested that the threads are not connected with the clubs, but are merely an adventitious micro-organism growing in the track of the ray-fungus. The threads are stained blue and the clubs crimson. In the younger clubs the protoplasm of the thread can be traced inta the interior of the club. In some of the older clubs the central portion takes a yellowish stain, and in others the protoplasm is not continued as a thread, but is collected into a spherical, ovoid, or pear-shaped mass. In others again, irregular grains, stained blue, are scattered throughout the central portion. The sheath of the thread is stained pink ; and the protoplasm, stained blue, fills the sheath, or consists of small spherical or irregular grains, giving a distinctly beaded appearance. The effect of various reagents should be tried upon the isolated 28 434 INFECTIVE DISEASES. grains. The grains are picked out of the pus and transferred to watch-glasses containing strong potash, xylol, and benzol. If returned to a slide and covered with a cover-glass, the clubs are found unaltered. Water or weak potash washes away the clubs, and the filaments become easily distinguished ; ether and strong acids have no effect upon them. Corallin soda, Hanstein's violet, and iodine zinc-chloride fail to give any particular reaction. Hoffman's blue stains the clubs, but without bringing out any structural details which could not be observed in the unstained specimens. Actinomyces bovis. — The fungus in cattle may in the same way be detected with the naked eye in the muco-purulent discharge, or in a scraping from the cut surface of a growth. The tufts of the fungus vary in size under different circumstances, from that of a grain of fine sand to that of a pin's head. If the pus or scraping be spread out on a slide and examined against a dark background, the grains appear to be white or yellowish-white in colour ; but if examined by transmitted light, they appear distinctly brownish. On pressing the cover- glass on the slide the grains readily flatten out, being of a soft, tallowy consistency ; or in the process of gently pressing the cover-glass on the slide with slight lateral movement, a distinct gritty sensation is transmitted to the finger, owing to the presence of calcareous matter. On examination with a low power the fungus will be recognised in the form of irregular patches scattered over the field, which might readily be regarded as collections of granular debris of a brownish or yellowish-brown colour, but on careful examination they are observed to have a more or less characteristic appearance. On examining with a higher power, spherical, ovoid, or reniform bodies are to be seen, which are either typical rosettes of clubs or granular masses, with here and there a club-shaped body at the periphery. Pus cells, round cells, fat granules, and minute spherical bodies may also be distinguished. If the grains consist of typical rosettes, and be merely covered with the cover-glass, and examined without being flattened out between the cover-glass and the slide, they will recall to mind, on focussing alternately the centre and the periphery, the appearance of the capituluin of a composite flower. The central portion appears to consist of spherical forms; these are the ex- tremities of the component elements, and as we focus the edge of the rosette these elements are seen laterally, and their characteristic club-form is readily distinguished. The central portion may be flattened against the cover-glass, and as the individual clubs vary DESCRIPTION OF PLATES XVII. AND XVIII. Actinomycosis Bovis. PLATE XVII. Section of an actinomycotic tongue stained by the method of Gram and with eosin. FIG. 1. — This illustrates the appearance which is usually seen under a low power, when a section is stained by Gram's method and with eosin. The central portion of a mass of the fungus is either unstained or tinged with eosin, while the marginal portion is stained blue. The reverse is seen, as a rule, in sections from man ; although under a low power the general appear- ance of sections from these two sources is somewhat similar, x 50. FIG. 2. —a, b, c> d, represent the earliest recognisable forms of the ray fungus in the interior of leucocytes. In e the club-forms can be recognised. In f and g there are small stellate groups of clubs, x 500. FIG. 3. — A part of the section represented in Fig. 1, under a high power. The marginal line of blue observed under a low power is now recognised as the result of the stain being limited to the peripherally arranged clubs. At («) part of a rosette has undergone calcification ; the clubs are granular, and have not retained the stain. At (J) and close to it there are the remains of rosettes in which the process of calcification is almost complete. x 500. PLATE XVIII. The figures in this plate are taken from sections of a case of so-called " osteosarcoma," in which the growth of the fungus was remarkably luxuriant. The specimens were stained by Plants' method. FIG. 1.— Different forms of clubs in different specimens : x 1200. O) Very small club-shaped elements. (&) A club with transverse segmentation, (c) A club with lateral daughter clubs. (d and ACTINOMYCOSIS. 435 considerably in size, the appearance of an irregular mosaic Is thus produced. By pressing upon the cover we break up the rosette, and then the clubs are recognised either singly or in pairs, or attached together in the form of wedge or fan-shaped segments. Calcareous material, if present, may readily be demonstrated by the action of acids. It will be found that on the addition of dilute hydrochloric, nitric or acetic acids, the calcareous deposit is dissolved while the clubs are not affected, and even with the addition of the strongest acids the only re"' it will be to dissolve out the calcareous matter and clarify the tutcs of the fungus, the form of the clubs being still recognisable. They are not affected by ether or potash, and thus the effects of chemical reagents clearly distinguish them from fat crystals or calcareous particles. By breaking up the growth into small frag- ments, we may readily study the shape of the individual club-like •elements. By using high-power objectives, and properly arranging the illumination, various forms will be clearly delineated. In some cases the club will be found to be bifid at the extremity ; in other cases there are lateral offshoots or daughter- clubs. Here and there will be found clubs closely pressed together like a bunch of bananas, and in other cases the broken-off pieces have a palmate form. By teasing out the grains in water, and pressing them apart between the slide and cover-glass, we find that the central portion is com- posed, as a rule, of a structureless core. More rarely there are the delicate filaments which are found in cultures and in the fungus from man. If the grains are mounted in glycerine, the appearance of the organism in the fresh state may be preserved. The granules may be stained by picking them out with needles and transferring them to a watch-glass containing alcohol, to which a few drops of concentrated alcoholic solution of eosin have been added. They remain in the solution until distinctly stained, and they are then placed on a glass slide in a drop of glycerine. The muco-pus may be spread out into as thin a film as possible on a cover-glass, allowed to dry, fixed by warming slightly over the flame, and stained by the method of Plant or of Gram. The characters of the fungus can be so readily recognised in the perfectly fresh state, that methods of staining are of secondary importance in diagnosis, though there are certain minute points which can only be satisfactorily determined by means of suitable dyes. 436 INFECTIVE DISEASES. .CULTIVATION OF ACTINOMYCES. Bostrb'm cultivated actinomyces from five cases in animals, and from one case in man. In all cases he obtained a similar result. The fungi were isolated from pus with sterilised needles, and placed in liquefied nutrient gelatine in which they were teased out, and the gelatine then spread on glass plates. The growth is stated to have become visible in a few days. The fungi were isolated from the plates, crushed between sterilised glass slides arid inoculated on the surface of nutrient agar-agar and blood serum. In this way pure cultivations were obtained. Nutrient gelatine was not liquefied. The cultures in blood serum and agar-agar grew best at from 33° to 37° C. The track of inoculation gradually spread out during the first two days, having a finely-granular, whitish appearance. During the next few days small yellowish-red spots appeared in the centre of the inoculated area, while the edge was apparently com- posed of fine processes. The yellowish spots continued to increase- for about seven or eight days, and became confluent, and the periphery also was dotted with yellowish-red points. Finally, there were also isolated colonies consisting of a yellowish-red centre with a greyish, downy periphery. The cultivated fungus, if suitably stained, corresponded exactly with that found in human and animal actmornycosis. In the cultures during the first two days threads were found, with true branchings ; later the threads were divided into shorter pieces or rods, and when the yellowish centres appeared there were also a number of very short rods and cocci- like forms. Bostrom also described attenuated club-shaped syell- ings at the end of the threads. He concluded by saying that Actinomyces is not one of the mould fungi, the central threads do not therefore constitute a mycelium ; he was inclined to regard it as a branched Cladothrix, and cultivation seemed to prove this. He suggested that it might be the Streptothrix Forsteri of Colin. Tii any case he relegated Actinomyces to the fission fungi or bacteria. In 1888 the author made cultures on glycerine-agar from a case of human actinomycosis of the thoracic wall. An abscess was opened, the discharge collected in sterilised tubes, and cultivations prepared with as little delay as possible. Some of the discharge was spread out on a sterilised glass slide, and the grains isolated with sterilised needles and quickly transplanted on the surface of the nutrient medium. The tubes were placed in the incubator at 37° C., and the result watched from day to day. For several days there was to the naked eye no promise of success; but gradually the grains began * ACTINOMYCOSIS. 437 to change, and by the end of a fortnight there was an appreciable increase in size. Numerous cover-glass preparations were made from what was originally a single grain, and on examination by the method of Gram the appearance was very striking. There could be no doubt as to the increase of the mycelial structure. The dense masses of filaments covered almost the whole area of the preparation. In parts less thickly covered there were very numerous oval bodies, and rod-like segments with terminal enlargements. These " crocus" forms corresponded with the appearances previously de- scribed as met with in the interior of certain clubs. From this it would appear that some other condition is necessary for the develop- ment of the fully formed club, which is the result of the sheath undergoing some change, possibly mucilaginous, resulting in the formation of a thick investment of the clubbed mass of protoplasm at the end of the thread. These club-shaped bodies represent organs of fructification, rather than the results of degeneration or death. The difficulty in accept- ing the view of their being entirely lifeless forms lies in the fact that the author has observed daughter-clubs growing from the mature clubs ; and, further, in the bovine fungus the author has been able to trace the stages in the development of a single club to a completely formed rosette. In the unstained condition, the clubs are found, on the whole, to be very regular in their form and arrangement, and by certain staining methods they can be shown to have a somewhat complex structure. If we take all the characters into account, and par- ticularly the minute structure and the relation to each other of the threads and clubs, we are justified in the opinion that the club in the early stages is an integral part of the living fungus, and that these characters bring the fungus into relation with a higher group of micro- fungi, Basidiomycetes, although the filaments, regarded by themselves, correspond with the characters of Streptothrix. The life-history of the micro-organism may be summed up thus : — The spores sprout into excessively fine, straight, or sinuous, and sometimes distinctly spirilliform threads, which branch irregularly and sometimes dichotomously. The extremities of the branches develop the club-shaped bodies. The clubs are closely packed to- gether, so that a more or less globular body is formed, with a central core composed of a dense mass of threads. The threads can be differentiated by the method of Gram into an external sheath, and protoplasmic contents. The club-shaped body externally appears to be mucilaginous, while internally it is continuous with the protoplasm I 438 INFECTIVE DISEASES. of the thread. It is difficult to say what further changes occur in the club-shaped bodies ; in all probability they represent organs of fructification. If so, the protoplasm in the interior of the club may possibly undergo changes leading to the development of spores, which are ultimately set free ; in some cases the terminal segment of a club is separated by transverse fission in the form of a globular body, a process resembling the formation of spores by abjunction. In others, the forms sprouting from the club are suggestive of teleutospores. There are occasionally long, slender forms, very different from the ordinary clubs ; they possibly represent paraphyses or abortive elements. In whatever way they may be formed, there can be little doubt that spores are set free in the vicinity of a rosette, and give rise to fresh individuals; the ultimate result recalling, as has been suggested, the appearance of " fairy rings." There can be little doubt that spores and young fungi are taken up by wandering cells, and conveyed to a distance from the parent fungus, and thus fresh centres of growth are established. Appearances of cultures. — Bostrom, Wolff, Israel, Paltauf, and others have shown that actinomyces can be cultivated in the ordinary nutrient media. More recently the author has carried on a series of cultivations for some years on glycerine agar, gelatine and milk, broth, bread-paste, and potato, in order to observe the changes which take place, and to study the variations which he found in the appearance of sub-cultures. The actinomyces after a few days on glycerine agar at the temperature of the blood forms little, white, shining, moist colonies, which may remain stationary, or increase and coalesce. In a week or ten days, sometimes earlier, and sometimes after several weeks, the cultures turn a bright yellow colour, but some remain, though white ; others, again, have a tinge of pink, and others are yellowish-brown (Plate XIX). After a time, a powdery efflorescence makes its appearance on the surface of the culture, which may be either yellow or white in colour. The culture may go on increasing, spreading over the surface of the medium, and retain its yellow colour, or it may turn black in parts or completely so, while the agar is coloured brownish-black. Cultures have a peculiar sour smell; the variations in cultures were all proved, by careful testing by sub-cultures, to be due to the growth of the actinomyces under varying conditions of soil, temperature, and the supply of air. The stage of efflorescence corresponds with the breaking up of the filaments into masses of cocci, and chains often closely resembling streptococci. Gelatine is slowly liquefied. Wolff and Israel cultivated actinomyces on raw and boiled DESCRIPTION OF PLATE XIX. Pure-cultivations of Actinomyces. These tubes were selected from a great number of cultivations in which there were different appearances. In some instances the growths had a faint tinge of pink. FIG. 1.— Pure-cultivation on the surface of potato, showing a luxuriant sulphur-yellow growth entirely composed of entangled masses of fila- ments. After three months' growth. FIG. 2. — Pure-culture from the same series, on glycerine-agar. In this case the culture remained perfectly white. The jelly was coloured reddish- brown. After fifteen months' growth. FIG. 3. — Pure-culture on glycerine-agar in which the growth was dark- brown, in parts black, and the jelly stained dark-brown. After nearly two years' growth. PlateHX. Figl. Fig 2. PURE- CULTIVATIONS OF ACT INOMYCES. Vbifgnl Bnolcs.Day £Son.,IitK. ACTINOMYCOSIS. 439 eggs, and succeeded, by inoculation in the peritoneal cavity, in producing the disease in rabbits and guinea-pigs, in the form of tumours of the peritoneum. Sauvageau and Radais consider that actinomyces should not be included with bacteria, and have suggested the name Oospora bovis. PREPARATION AXD EXAMINATION OF TISSUES. In order to examine the microscopical appearances, the tissues should be hardened in absolute alcohol and embedded in celloidin. The sections,, when stained, are to be dehydrated as a rule in strong spirit instead of absolute alcohol, as the latter dissolves the celloidin. If the sections are very friable, they can be cleared with clove-oil on the slide. By these means the little fungus tufts, which have a great tendency to fall out of the sections, may be preserved -in situ after passing through the various staining processes. To cut the sections, we can use either Jung's micro- tome, cutting in alcohol, or the freezing microtome. In the latter case, after the celloidin has hardened, it is necessary to shave off all that surrounds the piece of tissue. It is placed in water until it sinks, and then transferred to gum, and frozen and cut in the ordinary way. Staining Methods. There are several methods by which the organism can be stained in the tissues, but it is best to employ for this purpose Gram's method and modifications of Plant's method. Gram's Method. — By Gram's method the clubs in the bovine disease are distinctly stained, especially if the sections contain the fungus at a suitable stage. Use freshly prepared staining solution. A few drops of aniline-oil are placed in a test-tube, which is filled up with distilled water, the mouth of the tube closed with the thumb, and the mixture shaken up thoroughly. An emulsion forms, which is then filtered, until a perfectly clear solution of aniline-water is obtained. To this is added, drop by drop, an alcoholic solution of gentian-violet until precipitation commences. About fifteen to twenty drops in a small capsule of aniline- water will be sufficient. Sections are floated in this dye for about ten minutes, then transferred to the iodine-potassic-iodide solution until they turn brown like a tea-leaf. They are then decolorised in alcohol ; then stained in a weak alcoholic solution of eosin, dehydrated in strong com- mercial alcohol, cleared in clove-oil, and mounted in balsam. It will be found that the clubs are stained blue, and that there is a central area, which is, as a rule, tinged by the eosin. There are various modifications of the method, and some of them are extremely successful in affording not only a picture of the fungus, but also the structure of the surrounding tissue. Very instructive results may be obtained by combining the method of Gram with Ehrlich's histological stain. In this case, after the section has been decolorised in alcohol, it is ready to be transferred to logwood and treated as described below. Weigert's Method. — This also gives very beautiful results. The sections 440 INFECTIVE DISEASES. are placed for an hour in Wedl's solution of orseille, which is prepared as follows : — Add liquid extract of orseille to a mixture of absolute alcohol 20 parts, strong acetic acid 5 parts, distilled water 40 parts, until a dark-red liquid results. This must be filtered before use. The sections are left in this solution for an hour, then just rinsed in alcohol, and transferred to a solution of gentian-violet. Such sections show the nuclei of a violet-blue colour, and the peripheral part of the central core in the larger masses of the fungus also takes a blue colour, while the club-shaped structures are stained a striking wine-red colour. Plant's Method and Modifications. — This is one of the most valuable methods for staining the clubs. The original method was to float sections for ten minutes in magenta solution warmed to 45° C. This solution consisted of magenta two parts, aniline-oil 3 parts, alcohol of specific gravity 0'830, 20 parts, distilled water 20 parts (Gibbes). The sections were then rinsed in water, stained in concentrated alcoholic solution of picric acid for from five to ten minutes, immersed in water five minutes, 50 per cent, alcohol fifteen minutes, passed through absolute alcohol and clove-oil, and preserved in Canada balsam. The clubs are stained a brilliant red and the tissue yellow. Instead of employing the magenta solution, we now use Ziehl-Neelsen's solution. By removing the picric acid in Plant's method by prolonged immersion in alcohol, and then staining with gentian-violet or methylene-blue, a very successful contrast can be obtained. The most instructive histo- logical picture can be obtained by first staining with Neelsen's solution, removing the stain from the tissue in the way that has been already described, and then transferring the sections to distilled water, and subsequently staining with Ehrlich's histological stain. Ehrlictis New Histological Stain. — This is a combination of Ehrlich's logwood with orange-rubin. It is of especial value for sections of actinomycosis, and particularly in combination with carbolised fuchsine. It is employed in the following way :— The sections must be placed in alcohol or distilled water, and then in Ehrlich's logwood for about half a minute. From this solution they are transferred to distilled water, washed to remove the excess of stain, and then placed in a large dish of tap-water, where they are left for half an hour or more, until the sections turn blue ; if preferred, they may be left overnight. They are then stained for one or two minutes in a solution of rubin, S, and orange, and washed again in distilled water to remove the excess. They must then be dehydrated in alcohol, cleared in clove-oil, and mounted in balsam. Preparation of Large Sections. The method of cutting large sections of organs may be employed in studying actinomycosis. The value of these sections depends not only upon their affording an instructive picture of the naked-eye appearances, but they can also be studied with a pocket lens, or under the microscope with a | or £-in. objective. By staining the sections, the relation of the morbid to the healthy structures is brought out in greater contrast, and thus the topography of the disease can be studied more DESCRIPTION OF PLATES XX. AND XXI. Actinomycosis Bovis. PLATE XX. FIG. 1.— From a section of an actinomycotic tongue stained by the triple method (Ziehl-Neelsen, logwood and orange-rubin). In this section the separate centres of growth are clearly shown. Each neoplasm consists of a fungus system, in which the masses of the fungus, situated more or less centrally, are surrounded with round cells, epithelioid cells, sometimes giant cells, and lastly fibrous tissue forming a more or less distinct capsule. In parts the fungi have fallen out of the section. x 50. FIG. 2. — From a section of a "tubercular" nodule from the lungs of a Norfolk heifer with pulmonary actinomycosis. The nodule is a multiple growth surrounding a bronchus, and is enclosed by a capsule, in the vicinity of which the pulmonary alveoli are compressed. It is composed of a number of separate neoplasms, and each of the latter is composed of secondary centres of growth resembling the giant cell systems of bacillary tuberculosis. The new growth is composed of ray-fungi, large multi- nucleated cells, sometimes distinct giant cells, round cells, epithelioid cells, and, surrounding them, fibrous tissue. On examination of the same specimen with a higher power the typical rosettes of clubs are sometimes surrounded by multinucleated cells, and sometimes small rosettes are found like tubercle bacilli, in the interior of giant cells. From a pre- paration stained by Ziehl-Neelsen, logwood, and orange-rubin. x 50. PLATE XXI. FiG. 1. — (a) A leucocyte containing the fungus in its earliest recognisable form. (&) A large multinucleated cell containing the fungus in an early stage with the club-form already visible, (c) A leucocyte containing a small stellate fungus, (d) A large cell containing clubs arranged in a small rosette. (^^v>' •- "T^H'^wsw -^r ruwUc/eca. .CTINOMYCOSIS B' DESCRIPTION OF I'J I) XXI. Actinomycosis Bovia. PL A FlG. 1. — From a section of an acth tined by the triple method (Ziehl-Neelsen, logwo«> abin). In this section the separate centres of growth are i i neoplasm consists of a fungus system, in which the mass- s, situated more or less centrally, are surrounded lioid cells, sometimes giant cells, and lastly f more or less distinct capsule. In parts the i otlon. x 50. FIG. 2. — From a section of nodule from the lungs of a Norfolk heifer with puhn The nodule is a multiple growth surrounding »sed by a capsule, in the vicinity of which tin ompressed. It is composed of a number of separ .if the latter is composed of secondary centres of nt cell systems of bacillary tuberculosis. The new L of ray-fungi, large multi- nucleated cells. i -11s. round cells, epithelioid cells, and, surrounding r liination of the same specimen with a higher power ; if clubs are sometimes surrounded by innh M it-times small rosettes are found like tub- :->r of giant cells. From a pre- paration stained M :ued fuchsine. s. 1200. Lamplight illumination. Fit;. 2. — From a cover-glass preparation from the sam« source ; stained with -en's solution and methylene blue, x 1200. Lamplight illumination. Plate A 2. BACILLUS TETANI Brvolu.Day &Son,UlK. 460 INFECTIVE DISEASES. several months, and in rare cases a much longer period. The wound from a bite may have healed, and may again become inflamed, and symptoms follow, owing to the poison aft'ecting the brain, spinal cord, or the peripheral nerves. In the dog the disease appears in two forms —raging madness and dumb madness ; and the identity of the virus in the dog and in man is shown by the fact that virus from man can produce both forms of the disease in the dog. The virus may be obtained by inoculation of the saliva of a rabid dog ; but this method is uncertain, as other micro-organisms are present. Pasteur endeavoured to obtain it in a pure state, and was able to demonstrate that the spinal cord, the brain, and the nerves contain the virus. It was also found that by direct inoculation of the nervous system the most certain results followed. Bacteria in Rabies. — Cocci have been described in connection with hydrophobia by Fol, Babes, and Dowdeswell. The cocci, it is said, were observed in sections of the spinal cord of rabid dogs. The descriptions given by different observers vary considerably, and there is not any particular coccus constantly associated with the disease. Nor have any of the bacteria been isolated from the diseased animal, which were alleged to be present in stained preparations. By many, hydrophobia is believed to be due to the presence of a micro- organism, but at present the nature of the contagium is unknown. Protective Inoculation. — -Pasteur found that a dog inoculated under the dura- mater with virus from the spinal cord of a rabid animal will develop rabies, as a rule, within eighteen days. By trephining rabbits and inoculating the virus, and by, in the same way, transmitting the virus from rabbit to rabbit, the incubation period gradually shortens, until it is reduced to six or seven days. The virus has then reached its maximum virulence in the rabbit, and is " fixed." Pasteur then studied the possibility of producing immunity. The medulla of a rabbit, containing the virulent virus, was suspended in a, glass bottle over caustic potash at a temperature of 25° C. If a number of spinal cords were thus treated, and examined from day to day, it was found that they gradually lost their virulence, becoming completely inert in from sixteen to twenty days. A series of cords was thus obtained with diminishing virulence ; by injecting subcutaneously an infusion of rabid spinal cord crushed in broth, and beginning with an inert cord on the first day and using the next in the series on the second day, and so on till a fresh spinal cord could be injected, it was found that dogs were rendered insusceptible to the strongest virus, administered by inoculation or by exposing them to the bites of rabid dogs. Dogs have usually RABIES. 461 an incubation period of several weeks, and Pasteur conceived that it would be possible to anticipate the symptoms, which would naturally follow in a dog which had been bitten or inoculated, by giving them a mild form of hydrophobia by the injection of attenuated virus of short incubation period. These experiments showed that it was possible to do this, and the outcome was the introduction of a system of protective inoculation in the human subject. Pasteur succeeded in giving immunity from hydrophobia to about fifty dogs of every age and breed. In 1885 Joseph Meister, a boy nine years of age, bitten badly by a mad dog upon the hands, legs, and thighs, was brought to Pasteur. At a post-mortem examination of the dog, its stomach was found full of bits of hay, straw, and wood, and it had been unquestionably rabid. On July 6th, sixty hours after Meister had been bitten, a syringe full of marrow from a rabbit which had died on June 21st, and therefore fifteen days old, was injected beneath the skin over the right hypochondriac region. The next morning Meister was inoculated with a spinal cord fourteen days old, and so on every day, till on the sixteenth a cord only one day old was used. So many injections, however, need not have been given, as it was subsequently found that the spinal marrows injected during the first five days were inert when tested on rabbits. The marrows of the next five days showed an ascending scale of virulency, until, on the last two days of the treatment, Meister had been inoculated with a virus so virulent that it was capable of causing hydrophobia in dogs after ten days' incubation. Meister remained completely free from hydrophobia. From that time to the present day many thousands of patients have been treated in Paris by slightly modified methods, and it is very generally believed that a real prophylactic agent has been discovered. Pasteur suggested that the rabic virus might consist of two distinct substances — a living virus capable of developing in the nervous system, and a secondary product which, in sufficient pro- portions, might have the property of hindering the development of the living virus. The nature of this living virus is quite unknown. According to a return of the inoculations at the Pasteur In- stitute, the total number of persons treated in 1895 was 1,523, of whom five died. In three cases the symptoms of hydrophobia occurred within a fortnight of the last inoculation. If these three cases are omitted, the number of persons treated is reduced to 1,520 and the deaths to two. The results are shown in the following table,. 462 INFECTIVE DISEASES. for the nine years previous to 1895, during which Pasteur's method has been in operation : — Year. Number of persons inoculated. Number of deaths. Rate of Mortality. 1886 2,671 25 0-94 1887 1,770 14 0-79 1888 1,622 9 0-55 1889 1,830 7 0-38 1890 1,540 5 0-32 1891 1,559 4 0-25 1892 ... 1,790 4 0-22 1893 1,648 6 0-36 1894 ... 1,387 7 0-50 1895 ... 1,520 2 0-13 Of the 1,520 persons treated, 156 were bitten on the head or face, 829 upon the hands, and 535 on other parts of the body ; 122 were bitten by animals experimentally proved to be rabid, 949 by animals declared by veterinary certificate to be rabid, and 449 by animals supposed to be rabid. Babes at Bucharest inoculated 300 persons in one year, and claimed to have reduced the mortality to *4 per cent. Stamping-out System. — There is every reason to believe that rabies could be sta raped out in England in six months, if a general order for muzzling were enforced, and all ownerless dogs were slaughtered. It is the ownerless cur, the vagrant dog, which is mainly responsible for the spread of rabies ; and if a general muzzling order cannot be put into force, it would undoubtedly check the disease if all dogs were compelled to wear a collar with the name and address of the owner, and all dogs without owners were destroyed. LOUPING-ILL. Louping-ill is regarded by some as an infective disease. It is a disease of sheep, characterised by symptoms due to an affection of the central nervous system. The symptoms consist in contractions LOU PING-ILL. 463 of the muscles of the head and limbs, loss of co-ordination and finally complete loss of the power of movement. The name is derived from the peculiar jumping movements in the early stage. Klein and M'Fadyean independently investigated this disease. Klein found bacteria in the cerebral fluid. No micro-organisms were found in the blood. Special attention was drawn to a bacterium which was found by Klein in six out of seventeen cases, and to a micrococcus by M'Fadyean. Bacteria in Louping-Ill — Klein's Bacterium. Oval cocci and rods *6 to 1 /A in length, *2 to '3 /A in breadth. Colonies in gelatine, yellowish by reflected light, are brown by transmitted light. On the surface of gelatine the bacteria form a film, which is crenated at the edge, and thick in the middle, at first grey and later yellowish. In the depth of gelatine a filament forms, composed of closely aggregated minute greyish colonies, and a prominent yellow growth occurs on the free surface. The gelatine is not liquefied. The bacteria grow in milk, and broth becomes turbid in two days, and there is a copious flocculent greyish precipitate. Injection of broth cultures subcutaneously in rabbits, guinea- pigs, and mice, produced no result, except local swelling at the seat of inoculation, which subsided without causing any constitutional .symptoms. The results were equally negative when the cultures were injected subcutaneously in lambs. M'Fadyean' a Micrococcus. — Cocci '3 //, in diameter. The colonies are flat, nearly circular, and have a smooth edge. In old colonies the centre appears as a dark spot. Gelatine is rapidly liquefied, and a nearly colourless precipitate forms at the bottom of the tube. Cultures on the surface of agar have a faint yellow tinge. On potato the colour is deeper but the growth not so well marked. Milk is coagulated. In broth there is an abundant growth render- ing the liquid turbid and depositing a white precipitate. The micro- cocci stain by Gram's method. Inoculated in rabbits or guinea-pigs, they produce suppuration ; in horses and bovines, an inflammatory swelling results without suppuration. They produce abscesses in sheep and lambs. The cocci were isolated from abscesses in lambs suffering from louping-ill. Though it is admitted that louping-ill belongs to the class of infective diseases, there can be no doubt from these experiments that the nature of the contagium is unknown. CHAPTER XXXIII. FOOT-ROT. SHEEP are subject to several diseases which are classed as foot -rot. There is one form, known as contagious foot-rot, which prevails in certain localities, especially on wet land. Brown describes the disease as primarily a disease of the skin, inducing exfoliation of the cuticle, and exudation of fluid containing epithelial scales. The inflammation extends to the membrane of the foot, leading to exfolia- tion of the hoof, and development of epithelial scales, which form an imperfect horny layer on the diseased membrane. In one outbreak investigated by Brown, the disease in the early stage was confined to the skin between the digits of the fore-feet; the surface was red, tumid and pulpy, and white purulent matter existed on the inflamed parts. Later, the hoof grew to an extraordinary length, fungoid growths made their appearance, developing into foot-rot in an advanced form. In France, according to Fleming, the contagious character of this disease has long been recognised. So long ago as 1805 Pictet imported 200 half-bred merino sheep, some of which were infected with foot-rot, and placed them with 200 healthy sheep, and in a short time all the sheep were infected. Several years later Favre and Sorillon carried out investigations which conclusively proved the infective nature of the virus. Among other experiments it was found that when healthy sheep wrere inoculated in the feet with virus from diseased sheep, the disease was communicated. Contagious foot-root may be spread by healthy sheep receiving the virus from infected sheep in fairs and markets. Ships, railway- trucks, and carts in which diseased sheep have been conveyed, unless subsequently thoroughly disinfected, may be the means of trans- mitting the virus to healthy sheep. Healthy sheep turned into pastures quite recently occupied by diseased sheep may be inoculated from the discharges from the feet of the diseased sheep, which contaminate the grass and soil. 464 FOOT-ROT. 465 Law believed that the disease arose from an undiscovered micro- organism, which was probably present in infected pastures. Others in this country have disputed the contagious character of the disease, and considered that the same conditions of the pasture which produced the disease in a flock would produce it again in imported animals, which would account for the apparent contagiousness. Brown, in order to test the question of contagion, placed infected sheep in a pen, the bottom of which was covered with straw which was not removed while the experiments were in process. Three healthy sheep, from a locality where foot-rot was unknown, were placed with the infected sheep. At the end of ten days the feet of the sound sheep were still healthy. Subsequently two of the sheep were inoculated, and it was found that the virus introduced FIG. 187.— FOOT OP SHEEP SHOWING DISEASE OF HORN (BROWN). FIG. 188.— SECTION THROUGH THE FOOT SHOWING A CRACK EX- TENDING THROUGH THE WALL. subcutaneously in the vicinity of the foot produced the incipient stage of the disease. On making further experiments the contagious nature of one form of foot-rot was established, but it appears that the contagious property is only developed after a long period of exposure, and under certain conditions. On a dry soil the disease will quickly subside, but on moist land the contagious form of foot- rot may be communicated by simple contact, in from six weeks to three months. From these and other experiments Brown has drawn the following conclusions : — 1. That so far as the evidence goes it justifies the statement that foot-rot is a contagious disease ; the infective matter being active when brought into contact with the skin between the claws, or 30 466 INFECTIVE DISEASES. if II T OF THE ••riVERSlTY FOOT-ROT. 467 when introduced into the system by inoculation, and probably when taken in by the mouth from contaminated pastures. 2. That it cannot be produced by long-continued exposure to imdrained, moist soils, with an abundant, coarse and wet herbage. 3. That animals exposed to these conditions for many months, .and resisting entirely the influences named above, contract foot-rot FIG. 191.— DISTORTION OF HOOF IN AN ADVANCED FORM OF FOOT-ROT (BROWN). in from fourteen to twenty-one days on being placed among sheep suffering from the disease. 4. That sheep affected with foot -rot may improve, and from time to time become worse ; and finally may recover and present a perfectly healthy condition of foot, notwithstanding that they have been kept the whole period under the conditions which induced the disease. 468 INFECTIVE DISEASES. 5. That the contagium of foot-rot remains for some time in the system (ten to twenty days and longer) without any indication of disease appearing in the skin between the claws. An infected sheep may therefore escape detection even by an expert, and may introduce foot-rot into a sound flock. One attack does not confer immunity. The disease has been known to recur in sheep which have only recently recovered from an outbreak. The disease is no', communicable to other animals, including man. The flesh is harmless, but as in severe cases the sheep are emaciated, the carcass is in consequence of little, if of any, value as food. The nature of the contagium is unknown. Stamping-out System. — Sheep should not be allowed to be moved from an infected district except for slaughter. When sheep are purchased to add to the stock on a farm, they should be isolated for two or three weeks, and carefully watched before they are allowed to mix with other sheep. If this disease is detected in a flock, every animal should be carefully examined, and any suspicious as well as any diseased sheep should be completely isolated from the rest. Fleming recommends that those which have been in contact, though still apparently quite healthy, should as a pre- cautionary measure, be made to pass through a trough containing a solution of chloride of lime to a 'depth of about four inches. The solution is made by adding one or two pounds of chloride of lime to two buckets of rain-water. If the trough is placed at the entrance to the sheepfold, the sheep will be compelled to traverse it at least twice a day. It is also recommended, when diseased sheep are treated by this plan, that after recovery, all manure in the fold should be removed and destroyed, and the soil dug up to the depth of six inches, or lime freely spread over the surface. Troughs and hurdles must be thoroughly disinfected, and buildings freely ventilated after similar treatment. No locality can be con- sidered free from suspicion until one or two months have elapsed since the recovery of the last case CHAPTEE XXXIV. FOUL-BROOD—INFECTIOUS DISEASE OF BEES IN ITALY — PEBRINE — FLACHERIE — INFECTIOUS DISEASE OF CATERPILLARS. FOUL-BROOD IN BEES. FOUL-BROOD is a contagious disease attacking bees and especially the larvae. The larvae rapidly lose their healthy appearance, die and decompose, turning into a coffee-coloured mass. The cells of the honeycomb are mapped out by the dark- brown cappings of the cells FIG. 192.— DISEASED CQMB (COWAN). containing the diseased larvse. The decomposition is associated with the production of an unpleasant odour, which can be detected at some distance from an infected hive. The disease has been known from very early times. Preuss investigated it microscopically, and attributed it to " micrococci." These were really the spores of a bacillus, which was first observed by Cohn. Later, Cheshire and 469 470 INFECTIVE DISEASES. FIG. 193.— SPORES OF BACILLUS ALVEI. Cheyne investigated foul-brood, isolated the bacillus, and fully described its morphological and biological characters in nutrient media. By removing the cap of one of the diseased cells, the de- composing larva can be withdrawn, and cover-glass preparations will reveal delicate bacilli and large oval spores. The bacilli can be readily isolated and cultivated in nutrient gelatine. Bacillus alvei (Cheshire and Cheyne). Hods varying in size, and forming large oval spores. They are motile and possess flagella. Cultivated in nutrient gelatine in test-tubes, a delicate ramifying growth appears on the surface, and irregular whitish masses arise along the needle track. Pro- cesses shoot out from these masses, and extend through the gelatine for long distances. They are thickened at points in their course, and are clubbed at the ends. The gelatine is gradually liquefied, and the bacilli form a loose, white, floccu- lent deposit at the bottom of the tube. The liquid in the tube becomes yellowish in colour after a time, and gives off an odour of stale, but not ammoniacal, urine. On the surface of nutrient gelatine the bacilli grow out in chains of rods in single file, or of rows of several side by side. The processes which are formed tend to curve, and at a short distance from the track of the needle form a distinct circle, from which another process grows out, and a fresh circle is developed. The gelatine in the vicinity of the bacilli gradually liquefies, and channels are formed in the gelatine in which the bacilli move backwards and forwards. On nutrient agar-agar a whitish layer develops, consisting of bacilli arranged side by side, which in a few days are replaced by rows of spores similarly arranged. FIG 194.— PURE-CULTURE IN NUTRIENT GELATINE, x 4 (CHESHIRE AND CHEYNE). On INFECTIOUS DISEASE OF BEES IN ITALY. 471 potatoes they form a dryish, yellow layer, and in milk a tremulous jelly. A cultivation of the bacillus in milk, sprayed over a honey- comb containing a healthy brood of bee larvae, produced foul-brood. Adult bees fed on material containing bacilli became infected. Inoculation of mice and rabbits with the bacillus gave doubtful results. Stamping-out System.— The infected bees, combs, frames and quilts must be destroyed, and the hives thoroughly disirifected, as this is the only way in which the resistant spores can be got rid of. Cowan believes that if foul-brood were under Government inspection and infected hives were destroyed, the disease could be stamped out,. FIG. 195.— CULTIVATION ON THE SURFACE OF GELATINE, x 80 (CHESHIRE AND CHEYNE). INFECTIOUS DISEASE OF BEES IN ITALY. In Italy bees are subject to another infectious malady, and Canestrini has found bacilli in the bees and in the larvae, which are believed to be the cause of the malady. Bacillus of Infectious Disease of Bees (Canestrini). — Hods 2 /x in width and 4 to 6 /A in length, occurring singly, in pairs and in chains, sometimes capsulated. They are motile, and spore - formation is present. They liquefy gelatine, colouring the liquid pink and forming a white deposit. On agar they form a white growth, and on potato la claret-coloured layer. Cultures are said to be capable of producing the disease in bees and larvae. PEBRINE. The silkworm disease known in France as pebrine is characterised by the appearance of black patches on the skin of the worms. It was investigated by Cornalia, Nageli, and Pasteur. Pasteur's ~ . RA . OF THK UNIVERSITY 472 INFECTIVE DISEASES. prophylactic measures have been described in another chapter (p "<)• Panhistophyton ovatum. (Lebert. Nosema bombycis, Micro- coccus ovatus, Corpuscles du ver-a-soie). — Shining oval cocci, 2 to 3 /x long, 2 /x wide, singly and in pairs, or masses ; or rods, 2 '5 /x thick, and twice as long. They multiply by subdivision. They were experimentally proved to be the cause of pebrine, gattine, maladie des corpuscles or Flecksucht ; and were discovered in the organs of diseased silkworms, as well as in the pupae, moths, and eggs. Metchnikoff believes that these micro-organisms are not bacteria, but psorosperms. FLACHERIE. Silkworms are also subject to a very destructive disease known as jlacherie, flaccidezza, maladie de morts blancs. The worms cease feeding, die and become a putrid mass. The disease is dependent upon bad hygienic conditions, and is very infectious. The cause has not been determined with certainty, but it has been attributed to a streptococcus. Streptococcus bombycis (Mikrozyma bombycis, Bechamp).— Oval cocci 1-5 /x diarn., singly, in pairs, and in chains. They are said to be present in dust from infected localities. DISEASE OF CATERPILLARS. Forbes has described an infectious disease of the larvae of a caterpillar (Picris rapce). Cocci which were found singly and in masses, were regarded as the cause of the malady. PAET III. SYSTEMATIC AND DESCRIPTIVE. 473 CHAPTEK XXXV. CLASSIFICATION AND DESCRIPTION OF SPECIES. IN reviewing the history of the various classifications which have from time to time been proposed, we shall see that the gradual improvements in the means of studying such minute objects, the methods of cultivating them artificially, and of studying their chemistry and physiology, and the ever- increasing revelations of the microscope, have resulted in establishing these microscopic objects as members of the vegetable kingdom, ranking among the lowest forms of fungi, but with regard to the division into genera and species we are still in a position of doubt and uncertainty. Miiller, in 1773, was the first to suggest a classification. He established two genera, Monas and Yibrio, and grouped them with the Infusoria. In 1824 Bory de Saint Vincent also attempted a classification; but it was not until Ehrenberg in 1838, and Dujardin in 1841, worked at the subject, that a scientific distinction of species was attempted. Ehrenberg described four genera : — I. Bacterium . . filaments straight, rigid. II. Vibrio . . filaments snake-like, flexible. III. Spirillum . . filaments spiral, rigid. IV. Spirochaeta . . filaments spiral, flexible. Djijardin united Spirillum and Spirochseta, and classed them thus : — I. Bacterium . . filaments rigid, vacillating. II. Vibrio . . filaments flexible, undulatory. III. Spirillum . . filaments spiral, rotatory. Bacteria were still considered as Infusoria, but in 1852 Perty maintained that some of the smallest living organisms belonged to- the animal and others to the vegetable kingdom, and that Vibrios without question belonged to the latter. In 1853 Robin pointed out 475 476 SYSTEMATIC. the affinity of the Bacteria and Vibrios to Leptothrix ; and Davaine, in 1859, insisted that the Vibrios were vegetables, and were in fact allied to the Algse. Since that time a flood of light has poured in upon this subject through the writings of Hoffmann, Pasteur, Cohn, Rabenhorst, Hallier, Billroth, Warming, Nageli, Magnin, Marchand, Sternberg, Van Tieghem, Koch, Flugge, De Bary, Zopf, Biichner, Hueppe, Marshall Hall, and many others who have studied the morphology, life-history, and classification of bacteria. Of all these writers we are most indebted to Cohn, not only on account of his researches, which extended over many years, but also for his system of classification, which has since been almost iiniversally adopted. In his first classification, published in 1872, Cohn considered the Bacteria as a distinct group belonging to the Algse, and divisible into four tribes, including six genera : — I. Sphaerobacteria . . globules (Micrococcus). II. Microbacteria . . short rods (Bacterium). III. Desmobacteria . . long rods (Bacillus and Vibrio). IV. Spirobacteria . . spirals (Sphirochseta and Spirillum). Cohn noted, in spite of placing them with the Algae, that the absence of chlorophyll connected the Bacteria to Fungi, and we find Naegeli subsequently adopting this view, and employing the term Schizomycetes or Fission-fungi. Billroth, in 1874, disputed the division into species, and con- sidered that all the forms described by Cohn were but developmental forms of one micro-organism, Coccobacteria septica. In the following year Cohn answered the criticism of Billroth, and produced a second classification, in which he still maintained that distinct genera and species existed. Cohn considered the genera to be distinguished by definite differences in shape, which were adhered to throughout life, while some special feature, as a difference in size or physiological action, or some minute difference in form, determined the various .species. The second classification of Cohn (1875) only differed from the first in that, instead of keeping the bacteria as a separate group, he placed them, from their close relationship with the Phycochromacse, under a new group, the Schizophytes, and added the genera Lepto- thrix, Beggiatoa, Crenothrix, Sarcina, Ascococcus, Streptococcus, Myconostoc, and Streptothrix. Flugge retained the term Schizomycetes, and divided them thus : — CLASSIFICATION AND DESCRIPTION OF SPECIES. 477 SCHIZOMYCETES (FLtJGGE's OKIGINAL CLASSIFICATION). Isolated, or in chains, or united in amor- phous gelatinous families Micrococcus. Forming gelatinous families of definite form. Colonies solid, filled with cells Colonies, with simple layer of cells at the periphery . In large numbers, in irregular colonies . Ascococcus. In small but defi- nite numbers, in regular groups . Sarcina. Clathrocystis* Short. Isolated, or in small heaps loosely united, or in irregular gela- tinous families Short, distinctly jointed Long. Threads isolated, i n t e r -<; laced, or in bun- dles. Without ramifica- tions. Straight filaments. Long, ^ notdis-JThin tinctly j Thick jointed.' Wavy, or ( Short rigid Pseudo-ramifications . Bacterium. . Bacillus. Leptotlvrix . . Threads in roundish gelatinous masses Spirochffte (vibrio}. Spirillum. fStreptotltnj'. \Clathrotlirli-. . Myconostoc. 478 SYSTEMATIC. The belief is nevertheless rapidly gaming ground that the lowest forms of vegetable life cannot be divided by a hard-and-fast line into a series with chlorophyll (Algse), and a series without it (Fungi), and the tendency now is to solve the difference of opinion between Cohn and Nageli by following the example of Sachs, and amalga- mating the two series into one group, the Thallophytes. Researches by competent observers have more recently clearly demonstrated that several micro-organisms in their life cycle exhibit successively the shapes characteristic of the orders of Cohn. This doctrine of pleomorphism, now widely accepted, was dis- tinctly foreshadowed in a publication by Lister in 1873, though this memoir contained certain conclusions which have since been abandoned. Lister described forms of cocci, bacteria, bacilli, and streptothrix in milk, which he regarded as phases of the same micro-organism, Bacterium lactis. As a result of his observations, Lister remarks that " any classification of bacteria hitherto made from that of Ehrenberg to that of Cohn based upon absolute mor- phological characters is entirely untrustworthy." To Lankester, however, belongs the credit of having definitely and precisely formu- lated this doctrine. In a paper, also published in 1873, Lankester observed that the series of form-phases which he had discovered in the case of a peach-coloured bacterium led him to suppose that the natural species of these plants were " within the proper limits protean, and that the existence of true species of bacteria must be characterised, not by the simple form-features used by Cohn, but by the ensemble of their morphological and physiological properties as exhibited in their complete life- histories." Lankester inferred that these phase-forms were genetically connected, in that they all pos- sessed the common characteristic of a special pigment, bacterio- purpurin. These conclusions were vigorously opposed by Cohn, and doubt still remains in the minds of some as to whether the different forms are really only stages in the life-history of a single species. Nevertheless the theory of pleomorphism has steadily gained ground ever since. Cienkowski and Neelsen worked out the different forms assumed by the bacillus of blue milk ; Zopf has in a similar manner investi- gated Cladothrix, Beggiatoa, and Crenothrix, and traced out various forms (Fig. 196) ; Yan Tieghem has investigated Bacillus amylobacter with a similar result; Hauser has described bacillar, spirillar, spirulinar, and various other forms in the Proteus mirabilis and Proteus vulgaris. These facts obviously shake the very foundation of Cohn's classification, and we are left without possessing a sound CLASSIFICATION AND DESCRIPTION OF SPECIES. 479 basis for classification into genera or species. The mode of repro- duction is not sufficiently known to afford a better means for distinction than the other morphological appearances taken alone ; nor can we depend upon physiological action, which is held by many to vary with the change of form, according to altered surroundings. FIG. 196. CLADOTHRIX DICHOTOMA.— A. Branched Schizomycete : (a) Vibrio-form ; (b) Spiril- lum-form [slightly magnified]. B. Screw-form at the ends : (a) Spirillum- form ; (6) Vibrio-form. C. Very long Spirochseta-form. D. Branch fragment, at one end Spirillum-form, at the other Vibrio-form. E. Screw-form : (a) Continuous ; (6) Composed of rods ; and (c) Cocci. F. Spirochseta-form : (a) Continuous ; (6) Composed of long rods ; (c) Short rods ; and (d) Cocci (Zopf). Zopf, who has warmly supported the pleomorphism of bacteria, has suggested as a result of his investigations a division of the Schizomycetes, Spaltpilze, or Fission -fungi, into the following four groups : — 480 SYSTEMATIC. ZOPFS CLASSIFICATION. GROUP I. COCCACEJE. — Possessing (so far as our knowledge at present reaches) only cocci, and thread-forms resulting from the juxtaposition of cocci. The fission occurs in one or more directions. Genus I. Streptococcus (Chain-cocci). — Division in one or more direc- tions. Individual cocci remain united together to form chains. Genus II. Merismopedia (Plate-cocci). — Divisions in two directions, forming lamellae or plates. Genus III. Sarcina (Packet-cocci). — Division in three directions, form- ing colonies in cubes or packets. Genus IV. Micrococcus (Mass-cocci). — Division in one direction, cocci after division remain aggregated in irregular clusters, or singly, or in pairs or in chains of three or four elements. Genus Y. Ascococcus (Pellicle-cocci). — Like micrococcus, but the cocci grow in characteristic gelatinous pellicles. GROUP II. BACTERIACE^E. — Possessing mostly cocci, rods (straight or bent), and thread-forms (straight or spiral). The first may be absent, and the last possess no distinction between base and apex. Division (as far as is known) occurs only in one direction. Genus I. Bacterium. — Cocci and rods, or only rods, which are joined together to form threads. Spore-formation absent or unknown. Genus II. Spirillum. — Threads screw-form, made up of rods (long or short) only, or of rods and cocci. Spore-formation absent or unknown. Genus III. Leuconostoc. — Cocci and rods. Spore-formation present in cocci. Genus IV. Bacillus. — Cocci and rods, or rods only, forming straight or twisted threads. Spore-formation present either in rods or cocci. Genus V. Vibrio. — Threads screw-form in long or short links. Spore- formation present. Genus VI. Clostridium. — Same as bacillus, but spore-formation takes place in characteristically enlarged rods. GROUP III. LEPTOTRICHEJE. — Possessing cocci, rods, and thread- forms (which show a distinction between base and apex). The last straight or spiral. Genus I. Crenothrix. — Threads articulated ; cells sulphurless ; habitat water. Genus II. Beggiatoa. — Threads unarticulated ; cells with sulphur granules ; habitat water. Genus III. Phrngmidiothrix. — Threads jointless ; successive subdivision of cells is continuous ; cells sulphurless ; habitat water. Genus IV. Leptothrix. — Threads articulated or unarticulated ; successive subdivisions of cells not continuous ; cells sulphurless. GROUP IV. CLADOTRICH^EE. — Possessing cocci, rods, threads, and spirals. Thread-forms provided with false branchings. Genus :— Cladothrix. CLASSIFICATION AND DESCRIPTION OF SPECIES. 481 Zopf, however, does not assert that all the fission-fungi exhibit this pleornorphism, nor does he pretend that his classification will include all the micro-organisms described. Cohn, on the other hand, was ready to admit that all the forms described by him were not truly independent species. De Bary, Hueppe, Baumgarten, and Fliigge have expressed other \iews with regard to the classification of bacteria. De Bary divides them into two great groups — bacteria which form endospores, and bacteria which form arthrospores. This affords but little practical assistance, though regarded by botanists, from a scientific standpoint, as a step in the right direction. Hueppe, acknowledging that the fructification must eventually be made the basis for classification, suggests an arrangement for provisional use in which this view is introduced (p. 482). It has already been mentioned that the production of arthrospores is only established in a very few species. Therefore, we are hardly justified in assuming that all bacteria, the spore-formation of which is quite unknown, are to be included with those in which this kind of fructification has been observed, and consequently to distinguish genera on the same grounds may be considered, to say the least, somewhat premature. In Baumgarten's classification the genus bacterium is dispensed with, and the genera divided into two groups, the monomorphic and the pleomorphic. GROUP I. — MONOMORPHIC. Genera. — Coccus. Bacillus. Spirillum. GROUP II.— PLEOMORPHIC. Genera. — Spirulina. Leptothrix. Cladothrix. Fliigge also, in his revised classification, includes the genus bacterium in the genus bacillus. The new classification differs also from the original one in the grouping together of the different species according to the character and behaviour of the colonies in nutrient gelatine. The abolition, in Fliigge's and Baumgarten's classification, of the genus bacterium is no doubt owing to confusion having arisen from the distinction, between a bacterium and a bacillus, being made to depend upon length. Observers- differed as to whether a rod of a certain length ought to be considered a 31 482 SYSTEMATIC. CLASSIFICATION AND DESCRIPTION OF SPECIES. 483 bacterium or a bacillus. To meet this difficulty a rough-and- ready rule was suggested— viz., that a rod less than twice its breadth in length should be considered as a bacterium, and other- wise a bacillus. But this purely arbitrary division was inadequate, from the fact that a rod at one stage of its growth or under certain conditions might, as far as length went, truly be a bacterium, and under other circumstances be of such a length as to entitle its being considered a bacillus (Fig. 197). We should avoid such confusion if we followed Zopf, and acknowledged as a difference between a bacterium and a bacillus the presence or absence of that form of spore-formation now distinguished as endogenous spore -formation. We might then conveniently retain this generic term, to include that group of rod-forms in which this spore-formation is as yet FIG. 197. — FKIEDLANDER'S PNEUMOCOCCUS, x 1500 (Zopf). unknown ; moreover, we should, by so doing, with one or two exceptions, collect together those short rod-forms which appear to link the simple cocci to the spore-bearing rods or bacilli. The grouping together of the different species according to the character of the colonies in nutrient gelatine is also of questionable advisability. These characters can hardly be considered to be of sufficient importance, or indeed in many cases to be sufficiently constant, to serve by themselves for this purpose. In many cases a slight variation in the composition of the nutrient medium may considerably affect the appearances of the colonies. At the same time, the appearances are very characteristic of certain species of bacteria, and in some cases the characters of the colonies, together with the characters of the growth in test-tubes, assist us in distinguishing species which are morphologically similar, as in the case of the comma-bacilli of Tinkler and of Koch. 484 SYSTEMATIC. The classification of Zopf will lead the investigator to work upon the same lines, and by tracing the life-history of individual forms in pure-cultivations either to extend the work of establishing protean species or to restrict the doctrine of pleomorphism to a few forms. For though the author prefers the classification proposed by Zopf, he is not prepared to accept his views entirely— for instance, to regard the bacterium of rabbit septicaemia as a micrococcus. Any arrangement at present can only be considered provisional, and therefore the most practical classification must be considered the best. In fact, much more investigation is required before we can arrive at a permanent and thoroughly scientific classification of the known bacteria. Many bacteria have been described by different observers as different species which are really identical. Many micro-organisms have been described and named as new species with very insufficient investigation. The determination of species rests upon the accumulated evidence afforded by a thorough knowledge of their life-history. The morphological appearances under different conditions must be carefully studied, the presence or absence of movement and of spore-formation, and when present the exact character; the appearances of colonies and of test-tube cultivations in different media and under different circumstances ; the liquefaction and other changes in nutrient media ; the nature of the chemical products, if any, and the effect on the living animal of the bacterium itself and of its products, in varying doses, must all be taken into account. We must also ascertain whether the bacterium is an aerobe requiring the presence of oxygen, or an anaerobe growing only in the absence of it, or .a facultative anaerobe growing equally well with or without it ; and lastly, we must know whether the bacterium is a parasite requiring a living host, or a saprophyte existing on dead animal or vegetable matter, or a facultative parasite capable both of growing in the living animal and of leading a saprophytic existence. Several writers have classified the bacteria which have been described hitherto by taking some of these characters into account, and so preparing a list which is convenient for the purpose of bacteriological diagnosis. A system of this kind is of value in leading investigators to supply information which is wanting in order to verify and amplify the information upon which the classification is based, and to identify species which have been described under different names. CLASSIFICATION OF SPECIES FOE BACTEBIOLOGICAL DIAGNOSIS COCCI. (A) GELATINE NOT LIQUEFIED, (a) Chromogenic. COLOUB. HABITAT. Micrococcus violaceus . . . Violet . . . Water. Micrococcus carneus .... Flesh-red . . . Water. Micrococcus cerasinus siccus . . Cherry-red . . Water. Micrococcus cinnabareus . . . Cinnabar-red . . Air ; water. Micrococcus aurantiacus . . . Orange . . . Water. Micrococcus vecsicolor . . . Yellow ... . Water. Micrococcus luteus .... Yellow . . . Water. Micrococcus citreus .... Yellow . . . Water. Micrococcus ochroleucus . . . Yellow . . . Water. Micrococcus cereus flavus . . . Yellow . . . Water. Micrococcus agilis citreus . . . Yellow . . . Air. Micrococcus flavus tardigradus . . Yellow . . . Air ; water. Staphylococcus viridis flavescens . Greenish- yellow . Lymph. (b) Non-Chromogenic. Micrococcus candicans Micrococcus candidus Micrococcus concentricus . Micrococcus fervidosus Micrococcus cereus albus . Micrococcus aquatilis Micrococcus aquatilis invisibilis Micrococcus cumulatus tenuis . Micrococcus rosettaceus Micrococcus viticulosus Micrococcus plumosus Micrococcus amylivorus . Micrococcus acidi lactici . Micrococcus ureae Micrococcus gingivae pyogenes Micrococcus salivarius septicus Micrococcus of Manfredi . Micrococcus in pleuropneumonia Micrococcus in trachoma . Haematococcus bovis .... Diplococcus albicans tardissimus 485 Air ; water. Water. Water. Water. Pus; water. Water. Water. Nasal mucu>. Water. Air; water. Water. ' Pear-blight. Milk. Air; urine. Pus. Saliva. Sputum. Lymph. Conjunctiva. Urine. Vaginal secretion 486 CLASSIFICATION OF SPECIES Diplococcus coryzse Pseudo-diplococcus pneumonias Micrococcus tetragenus Micrococcus tetragenus mobilis ventriculi Pediococcus cerevisiae Pediococcus acidi lactici Streptococcus pyogenes Streptococcus brevis Streptococcus septicus Streptococcus vermiformis Streptococcus of mastitis in cows Streptococcus in strangles Streptococcus acidi lactici (B) GELATINE LIQUEFIED. (a) Chromogenic. COLOUR. Micrococcus agilis .... Pink . Micrococcus roseus .... Pink . Staphylococcus pyogenes aureus . Orange Micrococcus in pemphigus . . Orange Staphylococcus salivarius pyogenes . Orange Micrococcus fuscus .... Brown Micrococcus flavus desidens . . Yellowish-brown Micrococcus cremoides . . . Yellowish-white Micrococcus botryogenus . . . Yellowish. Micrococcus Finlayensis . . . Pale-yellow Staphylococcus pyogenes citreus . Yellow Micrococcus citreus liquefaciens . Yellow Micrococcus flavus liquefaciens . . Yellow Micrococcus tetragenus versatilis . Yellow Diplococcus flavus liquefaciens tardus Yellow Diplococcus subflavus . . . Yellow Diplococcus citreus conglomeratus . Yellow Diplococcus luteus .... Yellow Diplococcus roseus .... Pink . Diplococcus fluorescens fcetidus . Green (b) Non-Chromogenic. Micrococcus albus liquefaciens Micrococcus aerogenes Micrococcus radiatus Micrococcus fcetidus . . . . . . . Micrococcus in Biskra-button or Pendjeh sore Micrococcus in influenza . . . . . Micrococcus Freudenreichi ...... Micrococcus in yellow fever ...... Micrococcus lactis viscosus ...... Micrococcus acidi lactici liquefaciens .... Micrococcus ureae liquefaciens Micrococcus in gangrenous mastitis in sheep . Staphylococcus pyogenes albus Staphylococcus pyosepticus Diplococcus albicans amplus HABITAT. Nasal mucus. Meningitis. Sputum. Stomach. Beer ; air. Malt ; hay-dust. Pus. Saliva. Soil. Water. Pus. Pus. Milk. Water. Sputum. Pus. Bullaa. Saliva. Water. Air ; water. Water. Equine tumours. Yellow fever. Pus. Eczema. Air ; water. Blood. Eczema. Vaginal mucus. Pus ; air. Water. Air. Posterior nares. Nasal mucus. Intestine. Air ; water. Nasal mucus. Pus. Blood. Milk. Blood. Cream. Cheesy butter. Urine. Milk. Pus. Pus. Vaginal secretion. FOR BACTERIOLOGICAL DIAGNOSIS. 487 HABITAT. Pediococcus albus Water. Streptococcus liquefaciens Blood. Streptococcus septicus liquefaciens Blood. Streptococcus albus Water. Streptococcus of Mannaberg Urine. Streptococcus coli gracilis . Faeces. (c) NO GROWTH IN GELATINE. Micrococcus pneumonia crouposse Saliva. Micrococcus endocarditidis rugatus . . . . . . Heart. Nitromonas of Winogradsky ....... Soil. Micrococcus in pemphigus Bullae Micrococcus in influenza . . ... . . . Sputum. Micrococcus gonorrhoeas ........ Pus. Diplococcus intercellularis meningitidis Meningitis. Micrococcus tetragenus subflavus Nasal mucus. Streptococcus giganteus urethras . ; . . . . Urethra. Streptococcus of Bonome Meningitis. (D) GROWTH IN GELATINE UNDETERMINED. Ascococcus Bilrothii Meat infusion. Leuconostoc mesenteroides ....... Beet juice ; mo- lasses. Streptococcus of progressive tissue necrosis in mice . . Putrid blood. Micrococcus pyogenes tenuis Pus. Micrococcus of pyaemia in rabbits Meat infusion. Micrococcus of progressive abscess formation in mice . . Putrid blood. Micrococcus of Forbes Cabbage cater- pillars. Micrococcus in syphilis Blood Streptococcus Havaniensis Liver. Streptococcus perniciosus psittacorum Blood of parrot. Streptococcus bombycis Diseased silk- PACKET COCCI. (A) GELATINE NOT LIQUEFIED. (a) Non-Chromogenic. Sarcina pulmonum Phthisical sputum. Sarcina ventriculi Stomach. (B) GELATINE LIQUEFIED. (a) Chromogenic. COLOUR. Sarcina mobilis Red .... Ascitic fluid. Sarcina rosea Red .... Air. Sarcina flavea . Yellow . . . Beer. Sarcina lutea Yellow . . . Air. Sarcina aurantiaca .... Orange-yellow . . Air ; water. 488 CLASSIFICATION OF SPECIES (b) Non-Chromogenic. Sarcina alba Sareina Candida Air ; water. Air of breweries. RODS. (I.) AEROBES OR FACULTATIVE ANAEROBES (A) GELATINE NOT LIQUEFIED. (a) Chromogenic. (A) SPORE-FORMATION PRESENT. (jO) Motile. COLOUR. Bacillus cyanogenus Bacillus erythrosporus Bacillus brunneus Bacilhis rubefaciens Bacillus rubescens Bacillus fuscus limbatus Bacillus beroliniensis Indicus Bacillus cyanogenus Bacillus aurantiacus Bacillus fluorescens aureus Bacillus fluorescens longus . Bacillus fluorescens tenuis . Bacillus constrictus Bacillus subflavus Bacillus aureus . Bacillus flavescens Bacillus heminecrobiophilus Bacillus canalis parvus Bacillus fluorescens putidus Bacillus dentalis viridans Bacillus virescens Bacillus latericeus Bacillus spiniferus Bacillus in purpur; Bacillus uceus Bacillus in cholera in ducks Bacillus flavo-coriaceus Bacillus striatus flavis . Bacillus fuscus Greyish-blue . Blue milk. Greenish-yellow Water. iarrhcea (Lesage) Green Intestine. (V) Non-Motile. Brown Water. (B) SPORE-FORMATION UNKNOWN. O) Motile. Pale-pink Water. Pale-pink Sewers. us ... Brown Rotten eggs. [ndicus . . Indigo-blue Water. ordaniensis . Bluish Sewers. Orange Water. ireus . . Orange Water. ngus . . . Greyish-yellow Water. nuis . . . Greenish-yellow Water. Pale-yellow Water. Pale-yellow Water. Golden-yellow . Water ; eczema. . Yellow . Swamp- water. philus . . Yellowish Caseous glands. 3 ... Yellowish Sewer-water. itidus : . Greenish . Water. ans . . . Opalescent-green Carious dentine. Green Sputum. (J) Non-Motile. Brick-red Water. Greyish-yellow Eczema. emorrhagica . Greyish -yellow Blood. Orange-yellow . Water. ducks . . Yellowish Blood. is . . Sulphur-yellow Water. Sulphur-yellow Nasal mucus. Deep-yellow . Water. m-liquefaciens . Greenish -yellow Water. FOR BACTERIOLOGICAL DIAGNOSIS. 489 (b) Non-Chromogenic. (A) SPORE-FORMATION PRESENT. (a) Motile. Bacillus putrificus coli . . . .• . Bacillus septicus vesica . . ... Bacillus in cancer HABITAT. Human faeces. Cystitis. Stomach. Non-Motile. Bacillus acidi lactici (Hueppe) . Bacillus coprogenes foetidus Bacillus subtilis similans . Bacillus epidermidis . Bacillus of Colomiatti Sour milk. Swine measles. Human faeces. Skin. Conjunctiva. (B) SPORE-FORMATION UNKNOWN. (a) Motile. Bacillus oedematis aerobicus Bacillus of Fulles (I.) Bacillus stolonatus . Bacillus venenosus brevis . Bacillus venenosus Bacillus gracilis anaerobiescens Bacillus invisibilis Bacillus albus .... Bacillus venenosus invisibilis . Bacillus aquatilis sulcatus Bacillus argenteo-phosphorescens Bacillus gliscrogenus . Bacillus cystiformis . Bacillus of Guillebeau Bacterium Zopfii Bacillus ventriculi Bacillus coli communis Bacillus cavicida . . Bacillus of Utpadel . Bacillus aerogenes Helicobacterium aerogenes Bacterium aerogenes . Bacillus meningitidis purulentse Bacillus pyogenes fcetidus Proteus Zenkeri Bacillus enteritidis . Bacillus hyacinthi septicus Proteus lethalis .... Bacillus endocarditidis griseus . Bacillus of Roth (I.) . Bacillus Schafferi Bacillus of swine-plague . Bacillus typhi abdominalis Bacillus cavicida Havaniensis . Bacillus cuniculicida Havaniensis Earth. Earth. Water. Water. Water. Water. Water. Water. Water Water. Sea-water. Urine. Urine. Milk. Intestine ; air. Stomach of dogs. Intestine. Intestine. Intestine. Intestine. Intestine. Intestine. Pus. Pus. Putrid substances. Poisonous meat. Rotten hyacinths. Septicaemia. Endocarditis. Old rags. Cheese; potato. Lymphatic glands. Spleen and glands. Intestine. Intestine 490 CLASSIFICATION OF SPECIES (&) Non-Motile. HABITAT. Bacillus of Okada Dust. Bacillus pyogenes soli Earth. Bacillus of Fulles (II.) ........ Soil. Bacillus candicans . • Soil. Bacillus septicus agrigenus Manured soil. Bacillus scissus Soil. Bacillus ubiquitus Air ; water. Bacillus multipediculus . Air; water. Bacillus albus anaerobiescens . . . . . . . Water. Bacillus Zurnianus Water. Bacillus canalis capsulatus Sewer-water. Bacillus of Roth (II.) . . . Old rags. Bacillus tenuis sputigenus Sputum. Bacillus crassus sputigenus Sputum. Bacillus coprogenes parvus Faeces. Bacillus of Fiocca Saliva. Bacillus striatus albus . Nasal mucus. Bacillus capsulatus mucosus Nasal secretion. Bacillus pseudo-diphtheriticus Healthy throat. Bacterium ureas Urine. Bacillus nodosus parvus Healthy urethra. Bacillus oxytocus perniciosus Milk. Bacillus lactis pituitosi Milk. Bacillus limbatus acidi lactici Milk. Bacillus ovatus minutissimus Eczema. Bacillus of Belfanti and Pascarola Pus. Bacillus of Tommasoli Hair with sycosis. Bacillus capsulatus ......... Blood. Bacillus of purpura haemorrhagica (Babes) .... Blood, Bacillus of purpura haemorrhagica (Kolb) .... Blood. Bacillus septicaemiae haemorrhagicae ...... Blood. Proteus capsulatus septicus ....... Blood. Bacillus acidiformans Liver. Bacillus coli similis . Liver. Bacillus hepaticus fortuitus Liver. Bacillus filiformis Havaniensis Liver. Bacillus of Martinez ......... Liver. Bacillus diphtheriae columbrarum Diphtheritic de- posit. Bacillus diphtheriae Diphtheritic throat. Bacillus of Schimmelbusch Cancrum oris. Bacillus capsulatus Smithii Intestine. Bacillus erysipelatis suis Blood. Bacillus in rhinoscleroma Rhinoscleroma. Bacillus of Friedliinder Sputum. Bacillus pneumosepticus Septic pneumonia. Bacillus endocarditidis capsulatus Endocarditis. Bacillus septicus keratomalaciae Internal organs. Bacillus of intestinal diphtheria in rabbits .... Intestine. Bacillus of acne contagiosa of horses Pus. Bacillus pseudo-tuberculosis Internal organs. Bacterium tholcEideum Intestine. Bacillus gallinarum Blood. FOR BACTERIOLOGICAL DIAGNOSIS. 491 Bacillus argenteo-phosphorescens . Bacillus smaragdino-phosphorescens Bacillus phosphorescens gelidus Proteus hominis capsulatus Bacillus of grouse disease. Bacillus lactis aerogenes . HABITAT. Fish. Fish. Cuttlefish. Blood. Blood. Intestine. (A) GELATINE LIQUEFIED, (a) Chromogenic. (A) SPOKE-FORMATION PRESENT. O) Motile. COLOUR. Bacillus violaceus Deep-violet . . Water. Bacillus in disease of bees (Canestrini) Pink .... Larvae of bees. Bacillus in " red-cod " Red . . . . Salted codfish. Bacillus inesentericus ruber . . Reddish -yellow . Potatoes. Bacillus fluorescens liquefaciens minutissimus . . ... Greenish-yellow . Skin. (B) SPORE-FORMATION UNKNOWN. (a) Motile. Bacillus ianthinus . . . Bluish-violet . . Water. Bacillus violaceus Laurentius . Deep-violet . . Water. Bacillus lividus . . . . . . - Violet-black . . Water. Bacillus carnicolor .... Dark flesh -colour . Water. Bacillus rubidus Brownish-red . . Water. Bacillus Indicus Sealingwax-red . Intestine. Bacillus rosaceus metalloides Magenta-red . . Water. Bacillus ochraceus .... Yellow . . . Water. Bacillus citreus cadaveris . Yellow . . . Blood. Bacillus buccalis minutus . Yellow . . . Saliva. Bacillus arborescens .... Yellow . . . Water. Bacillus fulvus ..... Yellow . . . Water. Bacillus plicatilis .... Yellowish . . Water. Bacillus pyocyaneus .... Yellowish-green . Pus. Bacillus fluorescens liquefaciens Greenish-yellow\ . Water. Bacillus cyanofuscus .... Greenish -brown . Cheese ; glue. Bacillus fluorescens nivalis Bluish -green . . Water. Bacillus chromo-aromaticus Green or brown . Intestine. Bacillus viscosus. .... Green . . . Water. Bacillus pyocyaneus .... Green . . . Pus. (&) Non-Motile. Bacillus coeruleus .... Blue .... Water. Bacillus glaucus Grey. . . . Water. Bacillus membranaceus amethystinus Violet . . . Water. Bacillus lactis erythrogenes Red . . . . Milk. Bacillus mycoides roseus . Red .... Soil. 492 CLASSIFICATION OF SPECIES COLOUR. Bacillus prodigiosus .... Blood-red . Bacillus hydrophilus fuscus . . Yellow Bacillus cuticularis .... Yellow Bacillus helvolus .... Yellow Bacillus tremelloides .... Yellow Ascobacillus citreus .... Yellow Bacillus argenteo-phosphorescens liquefaciens Yellowish Bacterium termo (Vignal) . . . Yellowish Bacillus smaragdinus fcetidus . . Green Air. Frog's lymph. Water. Water. Water. Skin. Sea-water. Saliva. Ozaena. (b) Non-Chromogenic. (A) SPORE-FORMATION PRESENT. Motile. Bacillus inflatus . . . Urobacillus Freudenreichi Bacillus mycoides Bacillus ramosus . . Bacillus gracilis . . ., , Bacillus circulans Urobacillus Duclauxi Urobacillus Maddoxi Bacillus limosus .... Bacillus butyricus Bacillus Hessii .... Bacillus lactis albus . Bacillus liodermos Urobacillus Pasteuri . Bacillus meaentericus fuscus Bacillus mesentericus vulgatus . Bacillus of potato rot Bacillus maidis .... Bacillus megatherium Bacillus tumescens . Bacillus subtilis . Bacillus subtilis similis Bacillus vacuolosis . Bacillus of Scheurlen Bacillus alvei Air. Air ; dust ; sewers. Soil; water. Soil ; water. Water. Water. Water. Water. Sea-dredgings. Milk. Milk. Milk. Milk. Urine. Potato; dust; water. Potato; water, etc. Rotting potatoes. Maize infusion. Boiled cabbage. Beet-root. Dust ; water ; soil. Liver. Liver. Cancerous tissues. Larvae of bees. (b) Non-Motile. Bacillus aerophilus . Bacillus implexus Bacillus filiformis Bacillus vermicularis Bacillus incanus Bacillus inunctus Bacillus granulosus . Bacillus carotarum , Air. Water. Water. Water. Swamp-water. Swamp- water. Sea-dredgings. Boiled carrot. FOR BACTERIOLOGICAL DIAGNOSIS. 493 Bacillus brassicae Cabbage infusion. Bacillus in gangrene Senile gangrene. Bacillus of Letzerich ......... Urine. Bacillus anthracis Blood. (B) SPORE-FORMATION UNKNOWN, (a) Motile. Bacillus pestifer . . Air. Bacillus diffusus . ....... Soil. Bacillus gasoformans . . ... . . . Water. Bacillus liquidus Water. Bacillus guttatus . . . . '. Water. Bacillus liquefaciens . . . ". ; Water. Bacillus radiatus aquatilis . < . . . . . Water. Bacillus nubilis Water. Bacillus albus putidus . . . . . . . Water. Bacillus hyalinus . . Water. Bacillus vermiculosus .... . . . . Water. Bacillus dehcatulus . . . . . . . . . Water. Bacillus punctatus Water. Bacillus reticulans Water. Bacillus figurans (Vaughan) . Water. Urobacillus Schutzenbergi Water. Bacillus devorans Water. Bacillus venenosus liquefaciens Water. Bacillus aquatilis Water. Proteus sulphureus Water. Bacillus stoloniferus Swamp-water. Bacillus phosphorescens Indicus Sea-water. Bacillus phosphorescens indigenus Sea-water. Bacillus cyaneo-phosphorescens Sea-water. Bacillus litoralis Sea-dredgings. Bacillus halophilus Sea-dredgings. Bacillus superficialis Sewage. Bacillus cloacae . Sewage. Proteus microsepticus Uterine dis- charges. Proteus vulgaris .......... Putrid substances. Proteus mirabilis Putrid substances. Proteus septicus . Septicaemia. Bacillus foetidus ozsenae Nasal mucus. Bacillus septicus ulceris gangraenosi Blood and organs. Bacillus albus cadaveris Blood. Bacillus of Guillebeau Milk. Bacillus Havaniensis liquefaciens Skin. Bacillus carabiformis Stomach. Bacillus of Schou Rabbit. Bacillus leporis lethalis Liver. Bacillus liquefaciens communis Liver. (*) Non-Motile. Bacillus buccalis fortuitus Saliva. 494 CLASSIFICATION OF SPECIES HABITAT. Bacillus ulna (Vignal) Saliva. p Leptothrix buccalis (Vignal) Mouth. Bacillus varicosus conjunctive Conjunctiva. Bacillus gingivae pyogenes Alveolar abscess. Bacillus pulpse pyogenes . . Gangrenous tooth- pulp. Bacillus graveolens Skin of feet. Pneumo-bacillus liquefaciens bo vis Lung. (c) NO GROWTH IN GELATINE. (A) SPORE-FORMATION PRESENT. O) Motile. Bacillus ulna White of egg. Bacillus in putrid bronchitis Sputum. Bacillus mallei Glandered tissue. (ft) Non-Motile. Bacillus in erythema nodosum Blood. Bacillus tuberculosis Tubercular tissue, (B) SPORE-FORMATION ABSENT. Bacillus sanguinis typhi Blood. Bacillus septicus acuminatus Septic infection, Bacillus necrophorus Condyloma. (c) SPORE-FORMATION NOT STATED. Bacillus allantoides ^jr Bacillus nitrificans gojj Bacillus in measles Blood Bacillus in ophthalmia .... .'.'.' Conjunctiva. (D) GROWTH IN GELATINE NOT STATED. Bacillus senilis Blood. Bacillus leptosporus Air. Bacillus allii . . .' Putrefying onions. Bacillus mdigogenus Infusion of indigo. II. ANAEROBES. (A) GELATINE LIQUEFIED. (A) SPORE-FORMATION PRESENT. («) Motile. Bacillus rubellus Dust Bacillus butyricus (Botkin) ....'.'.' .' Milkjwater; dust Clostridmm fcetidum Earth FOR BACTERIOLOGICAL DIAGNOSIS. 495 HABITAT. Bacillus radiatus Earth. Bacillus liquefaciens magnus Earth. Bacillus spinosus Earth. Bacillus thalassophilus Sea-dredgings. Bacillus of symptomatic anthrax Tissues (quarter-ill). Bacillus cedematis maligni Lymph. Bacillus tetani Wounds ; earth. (&) Non-Motile. Bacillus liquefaciens parvus ....... Earth. Bacillus anaerobicus liquefaciens Yellow fever. (B) GELATINE NOT LIQUEFIED. (A) SPORE-FORMATION PRESENT. O) Motile. Bacillus amylozyma Water. Bacillus solidus . . Earth. Bacillus polypiformis Earth. (&) Non-Motile. Bacillus muscoides Earth. (B) SPORE-FORMATION ABSENT. (&) Non-Motile. Bacillus aerogenes capsulatus Blood. (c) GROWTH IN GELATINE NOT STATED. (A) SPORE-FORMATION PRESENT. (a) Motile. Bacillus butyricus Vegetable infu- sions, etc. (B) SPORE-FORMATION UNKNOWN. Bacillus cadaveris . *• Liver. CURVED RODS. (A) GELATINE NOT LIQUEFIED. (a) Chromogenic. O) Motile. COLOUR. Spirillum rubrum .... Deep-red . . . Putrid mouse. (b) Non-Chromogenic. Spirillum suis Intestine of swine. 496 CLASSIFICATION OF SPECIES Spirillum concentricum ... .... Putrefying blood. Spirillum saprophiles . . Hay -infusion; sewage. (i) Non- Motile. (a) Chromogenic. COLOUR. Spirillum flavescens .... Yellowish-green . Sewers. Spirillum flavum .... Ochre-yellow . . Sewers. Spirillum aureutn .... Orange-yellow . . Sewers. (b) Non-Chromogenic. Spirillum linguae Deposit on tongue. Spirillum nasale Nasal mucus. GELATINE LIQUEFIED, (a) Non-Chromogenic. (a) Motile. Spirillum cholerse Asiaticae . . " Intestine. Spirillum of Finkler and Prior Intestine. Spirillum Metchnikovi . . ... . . . Intestine of fowls. Spirillum of Miller Carious teebh. Spirillum of Sanarelli Water. Spirillum tyrogenum Old cheese. Spirillum marinum . . . Sea-dredgings. NO GROWTH IN GELATINE, OR UNDETERMINED. (a) Motile. Spirillum Obermeieri . . Blood. Spirillum anserum Blood of geese. Spirillum undula Putrid infusions. Spirillum sputigenum . . Gums. Spirillum serpens ......... Stagnant water. Spirillum tenue Putrid infusions. Spirillum volutans Swamp-water. Spirillum plicatile Swamp-water. (&) Non-Motile. Spirillum dentium Gums. Spirillum sanguineum . Brackish water. BRANCHING FILAMENTS. Streptothrix actinomycotica. Streptothrix alba. Streptothrix liquefaciens. Streptothrix musculorum suis. Streptothrix Hofmanni. Streptothrix farcinica. Streptothrix asteroides. Streptothrix carnea. FOR BACTERIOLOGICAL DIAGNOSIS. 497 Streptothrix aurantiaca. Streptothrix chromogenes. Streptothrix odorifera. Streptothrix violacea. Streptothrix Forsteri. Streptothrix madurae. NOT CLASSIFIED. Bacillus indigonaceus. Bacillus proteus fluorescens. Beggiatoa alba. Beggiatoa mirabilis. Beggiatoa roseo-persicina. Cladothrix dichotoma. Cladothrix invulnerabilis. Crenothrix Kuhniana. Diplococcus citreus liquefaciens. Leptothrix buccalis. Leptothrix gigantea. Micrococcus aquatilis invisibilis. Micrococcus crepusculum. Micrococcus foetidus. Micrococcus Havaniensis. Micrococcus of septicaemia in rabbits. Monas Okenii. Monas vinosa. Monas Warmingii. Myconostoc gregarium. Rhabdomonas rosea. Sarcina hyalina. Sarcina intestinalis. Sarcina litoralis. Sarcina Reitenbachii. Sarcina urinae. Sphaerotilus natans. Spirillum attenuatum. Spirillum leucomelaneum. Spirillum rosaceum. •Spirillum Rosenbergii. Spirillum rufum. Spiromonas Cohnii. Spimmonas volubilis. Streptococcus cadaveris. Streptococcus flavus desidens. Vibrio rugula. DESCRIPTION OF SPECIES AEEANGED FOE REFEBENCE IN ALPHABETICAL OBDEB. Ascococcus Billrothii.— Small globular cocci, united into charac- teristic colonies. They form on the surface of nourishing fluids a cream-like skin, divisible into an enormous number of globular or oval families. Each family is surrounded by a thick capsule of cartilaginous consistency. In a solution containing acid tar- trate of ammonia the fungi generate butyric acid, and change the origin- ally acid fluid into an alkaline one. FIG. 198. — Ascococcus BILLROTHII (Cohn). They were first observed on putrid broth, and later on ordinary nourish- ing solutions ; they also readily de- velop upon damp slices of boiled roots, carrots, beetroots, etc. Ascobacillus citreus (Unna, Tommasoli). — Rods sometimes curved, T3 p. in length, '3 p in width, singly, in pairs, and masses. The colonies develop slowly, and are yellowish in colour. The cocci inoculated in the depth of gelatine form small colonies in the track of the needle, and a slimv pale-yellow growth on the surface"; liquefaction sets in slowly. On agar the growth is gelatinous, and orange in colour, and rapidly extends over the surface. On potato the growth is abundant, and pale yellow. They were isolated from the skin in eczema seborrhoeicum. Bacillus acidiformans (Stern- berg). — Short rods, 1 -5 to 3 p in length, 1'2 /i in width, and filaments 5 to 10 p. Colonies circular ; iridescent by reflected light. Inoculated in the depth of gela- tine they grow freely in the track of the needle, and form a hemispherical mass on the surface. They produce gas bubbles. On agar the growth is milk-white, and the jelly becomes strongly acid. On potato the growth is abundant. In broth with 5 per cent, glycerine they produce opacity and a copious viscid deposit, and the surface is covered with gas bubbles. Injected into the peritoneal cavity of rabbits and guinea-pigs, they produce death in twenty-four hours. They were isolated from the liver in a fatal case of yellow fever. Bacillus acidi lactici (Hueppe). —Rods 1 to 2'8 /LI long, and '3 to '4 /x wide, and thread forms. Spore- formation present. In gelatine 498 DESCRIPTION OF SPECIES. cultures the breadth of the rods is diminished. They grow best be- tween 35° and 42° C., and cease under 10° C. Over 45'5° C. they no longer produce acidity. Whitish colonies appear on the second day. In gelatine a delicate growth appears along the whole track of the needle, with spherical forms here and there. In milk they produce lactic acid and the casein is precipitated. Bacillus aerogenes (Miller): — Small rods varying in length. Colonies white or yellowish- white ; concentric. In the depth of gelatine they produce a yellowish filament, and on the surface a grey patch with dentated periphery ; later the fila- ment is brown. On potato the growth is yellowish and dry. They were isolated from the in- testine in health. Bacillus aerogenes capsulatus (Welch). — Rods straight or slightly curved, 3 to 6 p. ; threads and chains ; capsulated. Colonies on agar greyish- white, with hairy processes. They peptonise gelatine and pro- duce gas. Broth becomes turbid, and there is an abundant sediment. Milk is coagulated. Cultures have a faint smell of glue. Injected into rabbits they pro- duce gas in the blood and internal organs. They were isolated from a patient after death, with blood-vessels full of gas. Bacillus aerophilus (Liborius). — Rods and filaments. Colonies punctiform ; greyish- yellow. Inoculated in the depth of gela- tine the bacilli produce a funnel of liquefied jelly, with flocculi in the lower part. On potato they form a smooth yellowish layer. They were isolated from con- taminated cultures. Bacillus albus (Eisenberg).— Rods and chains. Colonies circular, white. In gelatine the bacilli grow in the track of the needle, and form a white hemispherical mass on the free surface. On agar the growth is pure white, and on potato yellowish- white. They occur in water. Bacillus albus anaerobiescens (Vaughan). — Short rods. Colonies circular, yellowish- brown. Inoculated in the depth of gela- tine they grow in the track of the needle, and on the free surface. On agar the growth is pure white, and on potato yellowish-white. They occur in water. Bacillus albus cadaveris (Straussmann and Strieker). — Rods '2-5 n in length, '75 /z in width, and filaments. Colonies yellowish ; circular, and later radiated. Inoculated in the depth of gela- tine they produce a funnel of lique- fied gelatine with a thick deposit. On agar there is an abundant white growth. On potato the growth is white or yellowish-white, and colours the potato in the vicinity bluish-brown. The cultures have a putrefactive odour. Mice inoculated subcutaneously die in six hours, and guinea-pigs in twenty-four. They were isolated from putrid human blood. Bacillus albus putidus (De Bary). — Rods and filaments. -:J Colonies circular and brownish. Inoculated in the depth of gela- tine they produce rapid lique- faction. On agar and potato the growth is slimy. Cultures develop a strong putrefactive odour. They occur in water. Bacillus allantoides (L. Klein), — Rods 2 to 2*5 /u in length, '5 p. in width, and in chains. The rods develop cocci-forms united by a gelatinous substance into zooglcea masses. They were isolated from a contaminated culture. Bacillus allii (Griffiths).— Rods oOO DESCRIPTION OF SPECIES. 5 to 7 p in length, 2 -5 /x in width, singly, in pairs, and zoogloea. On agar they produce a bright green film, and cultures are said to emit traces of sulphuretted hydro- gen. They were isolated from putrid onions. Bacillus alvei (p. 470). Bacillus amylozyma (Perdrix). — Rods 2 to 3 /u, in length and "5 p in width, in. pairs, and in chains. They are anaerobic. Colonies white, and producing gas bubbles. On potato in an atmosphere of hydrogen the bacilli partly liquefy it, and there is abundant formation of gas. They ferment sugar and starch. Bacillus anaerobicus lique- faciens (Sternberg). — Slender rods, about '6 n in diam., in pairs, and in filaments. Colonies granular and white ; sur- rounded by liquefied gelatine. They grow along the track of the needle when inoculated in the depth of agar. They were isolated from the in- testine in a fatal case of yellow fever. Bacillus anthracis (p. 192). Bacillus aquatilis (Frankland). — Rods 2 -5 [JL in length, and filaments 17 fj. or longer. They resemble Bacillus arborescens. Colonies after liquefaction of the gelatine have a yellowish-brown nucleus from which proceed twisted strands of filaments. Inoculated in the depth of gela- tine the growth in the track of the needle is at first almost invisible, later liquefaction occurs. On agar the growth is shining and yellowish. Broth becomes turbid, and a sediment forms at the bottom of the tube. On potato there is a slightly yellowish streak. They occur in water. Bacillus aquatilis fluorescens (Lustig). — Short thin rods with rounded ends. Non-motile. Colonies fern-like and iridescent. Compare Eisenberg's Bacillus fluo- rcscens non-liqiiefaciens. Bacillus aquatilis graveolens (Tataroff). — Slender rods 1-3 /A in length. They rapidly liquefy gela- tine and produce an odour like that of perspiration from the feet. They occur in water. Bacillus aquatilis sulcatus (Weichselbaum), No. I. — Rods mor- phologically, and in cultures resem- bling Bacillus typhosus. Colonies in gelatine exhibit lines and furrows. The growth on the surface of gelatine is said to be greater than in cultures of Bacillus typhosus grown for comparison. They occur in water. No. II. — Rods also resembling in morphology and cultivation the Bacillus typhosus. Colonies are said to be thicker than those of No. I., and not dentated. The growth on potatoes is yel- lowish-brown, and emits a faint odour of urine. They occur in water. No. III. — Very short rods. Colonies show lines and furrows, and are yellowish. On the surface of gelatine the growth develops as a thin whitish film. On agar the growth is white and abundant. On potato the growth is yellow. They were isolated from water. No. IV. — Rods and filaments. Colonies circular and bluish. On the surface of gelatine the growth is greyish-white, and on agar there is a similar appearance. They do not grow on potato. They occur in water. No. V. — Rods rather thicker than those of Bacillus typhosus. Colonies similar to those of No. I. The growth on the surface of gelatine is yellow. On agar the growth is viscid and yellow, and on potato the growth is faintly yellow and the surrounding medium stained bluish-grey. They occur in water. Bacillus arborescens (Frank- land). — Rods 2'5 p in length, and DESCRIPTION OF SPECIES. 501 •5 p. in width, singly, in pairs, and in short chains, and filaments. Colonies throw out delicate branches with a highly characteristic appearance ; the gelatine slowly liquefies, the nucleus of the colony becomes yellow, and the periphery iridescent. . Inoculated in the depth of gela- tine the bacilli form a cloudiness in the track of the needle, and an iridescent layer on the surface with central depression of the gelatine and commencing liquefaction. Later the liquefaction produces a funnel, and there is a yellow deposit. On the surface of agar the layer is a dirty orange colour. On potato the growth is orange red with irregular protuberances, and limited in growth. They occur in water. Bacillus argenteo-phosphores- cens (Katz), No. I.— Rods slightly curved with pointed ends, 2'5 ^ in length, width one-third of their length ; singly, in pairs, and long wavy filaments. Colonies circular ; at first trans- parent droplets, later yellowish in colour. On the surface of gelatine they form a greenish-yellow film. In broth they produce turbidity, and later a skin on the surface, and on sterilised fish a pale-yellow sticky growth. Cultures are photogenic. They were isolated from the sea at Sydney. No. II. — Rods with rounded ends •27 /A in length, '67 /x in width, and filaments. Colonies on gelatine are circular with sharp contours and greyish- yellow in colour ; later they are irregular and granular. Inoculated in gelatine the bacilli form a greyish-white filament in the track of the needle, and a shining patch on the surface. On the surface of obliquely solidi- fied gelatine they form a bluish- grey film. In broth they produce only turbidity. Cultures are photogenic. They were isolated from phospho- rescent fish. No. III. — Rods not so thick as those of No. II., singly, in pairs, and filaments. They are motile. Colonies are white, scaly, and wrinkled. On the surface of gelatine the growth spreads over the medium. On agar the growth is scanty. In broth they produce turbidity and a skin floating on the surface. Cultures are photogenic after a few days' growth. They were isolated from a piece of cuttle-fish. Bacillus argenteo-phosphores- cens liquefaciens.— Rods straight or slightly bent, 2 ^ in length, and in width one-third of their length ; filaments. Colonies circular, pale brown or pale yellow and, after liquefaction, with radiating processes extending into the surrounding gelatine. Inoculated in the depth of gela- tine there is a growth in the track of the needle, and near the surface a cup-shaped area of liquefaction. In broth they produce turbidity, and form a skin on the surface. On sterilised fish they form a yellow layer. They are photogenic but not markedly so. Bacillus aurantiacus (Frank- land). — Rods short and thick, singly, in pairs, and in filaments. Colonies are prominent and pale orange in colour. Inoculated in the depth of gela- tine there is a slight growth in the track of the needle and an orange patch on the free surface. On agar and potato the growth is also orange. They occur in water. Bacillus aureus (Adametz).— Slender rods straight or slightly bent, 1'5 to 4 p. in length, and '5 p. in width ; in pairs, filaments, and masses. They are motile. Colonies circular or oval and yellow in colour. Inoculated in the depth of gela- tine the growth is very limited in DESCRIPTION OF SPECIES. the track of the needle, while small chrome-yellow hemispherical masses develop on the free surface. On potato they form a chrome- yellow growth. They occur in water and on the skin. Bacillus berolinensis Indicus (Classen). — Slender rods, singly, in pairs, and short chains ; capsulated. Colonies at first whitish acquire in a few days an indigo-blue colour. On the surface of gelatine they form a blue layer, which slowly spreads. On the surface of agar the indigo- blue colour is very marked. On potato they grow abundantly, and develop the same colour. The pigment is insoluble in alcohol, chloroform or water, soluble in strong acids, and decolorised by ammonia. They were isolated from river water at Berlin. Bacillus brassicae (Pommer).— Rods 1-9 to 5-4 n in length, -91 to 1'2 /LI in width, and filaments. They form spores. Colonies have the appearance of a fine mycelium. Inoculated in the depth of gela- tine the growth in the track of the needle sends off fine filaments, and liquefaction quickly follows. In the depth of agar white colonies form in the track of the needle, and on the surface the growth is first cloudy and later yellowish. They were isolated from infusion of cabbage. Bacillus brevis (Mori).— Rods •25 p. long and -8 p broad. Non- motile. Colonies pale-yellow ; non-lique- fying. Inoculated in the depth of gela- tine small dots appear along the needle track and a pale yellowish growth on the surface. On agar at 35° C. a yellowish and on blood serum a greyish growth appears in two or three days. They do not grow on potato. In broth they form a white cloudy -deposit. Mice inoculated subcutaneously die in from sixteen to thirty hours. They are also pathogenic in guinea- pigs and rabbits. They were found in drain water. Bacillus brunneus (Adametz and Wichman). — Rods small and slender. Spore formation present. Colonies at first white, later brownish. Inoculated in the depth of gela- tine the growth occurs along the track of the needle and also on the surface, developing a brownish colour in the surrounding gelatine. They occur in water. Bacillus buccalis fortuitus (Vignal).— Rods 1-4 to 3 /u in length, singly, and in pairs. Colonies circular and liquefying. Inoculated in the depth of gela- tine liquefaction occurs slowly, and white flocculi occur in the liquid, and later subside to the bottom. In broth they produce turbidity and a skin on the surface. They were isolated from saliva. Bacillus buccalis maximus (Miller).— Rods 2 to 10 p in length, 1 to 1-3 fjt. in width, and filaments 30 to 150 /n in length. They occur in the mouth. Bacillus buccalis minutus (Vig- nal).— Rods -5 to 1 /ji. in length, and slightly less in width. Colonies circular and faintly yellow. Inoculated in the depth of gela- tine they form a yellowish-white growth in the track of the needle, and a patch of the same colour on the surface ; liquefaction com- mences slowly, and extends down- wards until the gelatine is com- pletely liquefied and a yellow mass collects at the bottom of the tube. In broth there is a similar deposit, and an iridescent pellicle on the surface. On potato they form a yellow film. They were isolated from saliva. Bacillus butyricus, (Praz- mowski. Bacilhi,* amylobacter, Van Tieghem ; Bacillus of butyric acid fermentation}. — Rods 3 to 10 ft long, under 1 /* wide, often indistinguish- able from Bacillus subtilis. They DESCRIPTION OF SPECIES. 503 grow out into long, apparently un- jointed threads. They are mostly actively motile, but also occur in zoogloea. The rods and threads are sometimes slightly bent, like vibrios. They are anaerobic. The shorter rods as a rule swell in the middle, becoming ellipsoidal, lemon, FIG. 199.— CLOSTRIDIUM BUTTKICUM. A. Active stage, (a, 6) Bent rods (vibrio- form) and threads. (c) Short rods, (d) Long rods. B. Spore-formation. C. Spore-germina- tion. (Prazmowski. ) or spindle-shaped ; the long rods, ; and sometimes the short ones, swell ! at one end ; in either case ellipsoidal ! spores are developed (Fig. 199). Cultivated in nutrient gelatine, j the medium is liquefied, and a scum j formed on the surface. They grow [ best between 35° and 40° C. The spores are widely distributed in nature, and grow readily on fleshy roots, old cheese, etc. They convert the lactic acid in milk into butyric acid, and produce the ripening of ! cheese. They occur also in solutions of starch, dextrine, and sugar, and are the active agents in the fermenta- tion of sauerkraut and sour gherkins. Bacillus foutyricus (Botkin).— Rods and filaments, spore -formation present. They are anaerobic. Colonies consist of felt-like masses. Inoculated in the depth of gela- tine with 1-5 per cent, of grape- sugar, the growth commences in the lower part of the needle track with abundant formation of gas bubbles and liquefaction of the jelly. In milk there is abundant gas formation, which will break the flasks if closed. They were isolated from milk, earth, and water. Bacillus butyricus n (Hueppe). — Rods slightly bent, * 2-1 p in length, -38 /* in width, and filaments. Colonies yellowish ; rapidly liquefying. Inoculated in the depth of gelatine liquefaction occurs along the track of the needle, and later a wrinkled skin floats on the surface. On agar the growth is yel- lowish. On potato the growth is wrinkled and faintly yellow. They coagulate milk, precipita • ting and then dissolving the casern. They occur in milk. Bacillus cadaveris (Sternberg). — Rods 1 '5 to 4 /A in length, and 1*2 /LI in width, singly, in pairs, and in filaments. They are anaerobic. Colonies in glycerine-agar are irregular, granu- lar, and white. They produce an acid reaction in cultures. Subcutaneous injection in guinea- pigs may produce extensive 504 DESCRIPTION OF SPECIES. oedema and death in twenty-four hours. They were obtained from the liver in fatal cases of yellow fever. Bacillus caeruleus (Smith).— Rods 2 to 2-5 \i in length, and '5 /* in width, singly and in chains. Colonies blue. Inoculated in the depth of gela- tine they form a colourless growth in the track of the needle, and a cup-shaped cavity in its upper part, with bluish contents. On agar they form a blue layer and a deep-blue growth on potato. They occur in water. Bacillus canalis capsulatus (Mori).— Rods '9 to 1*6 p. in width, i capsulated. Colonies milk-white. The growth in the depth of gela- tine is similar to Friedlander's pneumococcus. On agar the growth is viscid, and on potato it is yellowish. In broth a skin forms on the surface. They are pathogenic in mice. They occur in sewage. Bacillus canalis parvus (Mori). -Rods 2 to 5 p. in length, -8 to 1 p in width. Colonies very minute, pale yellow. On the surface of gelatine they very slowly form a yellowish film. On agar the growth is dry and yellow. They are pathogenic in mice and guinea-pigs. They occur in sewage. Bacillus candicans (Frankland). — Very short rods and filaments. Colonies pure white. Inoculated in the depth of gela- tine they form isolated colonies in the track of the needle, and a white button on the free surface. On agar they form a greyish layer, and flourish on potato. They occur in soil. Bacillus capsulatus (Mori). — Oval forms and rods, sometimes encapsuled. Non-motile. Colonies white. Inoculated in the depth of gela- tine and agar a nail-shaped growth occurs. In broth they form a white- turbidity, and a white pellicle develops on the surface and on the sides of the vessels. On potato at 37° C. an abundant moist, yellowish, stringy growth is formed, with production of gas bubbles. They are pathogenic in mice and in rabbits if injected into the pleural cavity. They occur in drain water. Bacillus capsulatus (Pfeiffer). - — Rods singly, in pairs, or in chains and in filaments. They have a well- marked capsule. Colonies white. Inoculated in gelatine they grow in the track of the needle, and form a white button on the free surface. On agar and on potato the growth is also white and very viscid, so that it can be drawn out into long threads. They produce a fatal result in mice in two or three days, when inoculated subcutaneously. A minute quantity of a broth culture injected into the peritoneal cavity of guinea-pigs will prove fatal in thirty-six hours. The bacilli are found in the blood which is made viscid. They were isolated from a guinea- pig found dead. Bacillus capsulatus mucosits (Fasching). — Rods 3 to 4 p in length, '75 to 1 ju, in width ; capsu- lated. Colonies white. Cultures in gelatine resemble Friedlander's pneumococcus. They form gas. They produce a fatal result in mice in thirty- six hours. They wrere isolated from nasal mucus in cases of influenza. Bacillus capsulatus suis (Smith).— Rods from 1-2 to 1'8 /z in length and '8 to '9 /j. in width. There are three varieties of this bacillus having a close resemblance to the pneumococcus of Friedliinder. They were isolated from the in- testines of swine. Bacillus carabiformis (Kac- zynsky). — Rods short and slender. Colonies develop characteristic processes. DESCRIPTION OF SPECIES. 505 Inoculated in the depth of gela- tine, the bacilli produce liquefaction and colour the liquid greenish- yellow. On agar they form a yellowish- white layer. They were isolated from the stomach of a dog. Bacillus carnicolor (Tils).— Rods 2 ft long, and *5 ft broad. Singly actively motile. Spore- formation not observed. Colonies are in the form of cup- shaped depressions. Inoculated in the depth of gela- tine they grow rapidly along the whole track of the needle, forming a funnel-shaped area of liquefaction at the bottom of which there is a pale pink deposit. On potato they form slowly a dark flesh-coloured growth. They occur in water. Bacillus carotarum (A. Koch). —Rods -97 to 1'05 p. in length and filaments. Colonies white and circular. Inoculated in the depth of gela- tine the bacilli grow slightly in the track of the needle and abundantly on the surface. On agar they form a white, and on potato a faintly brown layer. They occur on boiled carrot and beet. Bacillus cavicida (Brieger). — Rods morphologically and in culti- vations similar to Bacillus coli com- munis. Cultures are said to be patho- genic in guinea-pigs. They were isolated from fseces. Bacillus cavicida Havaniensis (Sternberg). — Rods 2 to 3 ft in length, and •? ft in width, singly and in pairs. Colonies are of a pale-straw colour. Inoculated in the depth of gela- tine the bacilli form small trans- lucent pearl-like spherical colonies, and on the free surface the growth is limited. On potato the growth is at first thin and dirty yellow, and later gamboge yellow. Guinea-pigs inoculated subcuta- neously die in ten or twelve hours. They were isolated from the intestinal contents in a fatal case of yellow fever, by inoculation of guinea-pigs. Bacillus chromo-aromaticus.— Rods which liquefy gelatine and form a yellowish-white scum on the surface. On potato the growth is irides cent and brownish. In broth a scum forms on the surface and the broth is coloured greenish-blue. Cultures have an aromatic odour. They are said to produce pneu- monia and pleurisy in rabbits. They were isolated from a pig with postmortem appearances of swine-fever. Bacillus circulans (Jordan). — Rods 2 to 5 fi in length and 1 ft in width, singly and in short chains. Colonies are brownish. Inoculated in the depth of gela- tine they liquefy the medium in the track of the needle, forming a conical cavity at its upper part. On agar they form a translucent film. Milk is slowly coagulated. In broth they produce turbidity and a slimy deposit. They occur in water. Bacillus citreus cadaveris (Strassmann). — Rods '9 ft in length, •6 ft in width, singly and in chains. Colonies pale yellow. Inoculated in the depth of gela- tine the bacilli form minute colonies along the track of the needle, and at its upper part liquefy the gela- tine and produce a yellow deposit. They were found in the blood after death. Bacillus cloacae (Jordan).— Short rods '8 to 1-9 ft in length, •7 to 1 ^ in width, singly and in pairs. Colonies circular, yellowish. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle, an iridescent scum forms on the surface, and there is. an abundant deposit. On agar the growth is milk- 306 DESCRIPTION OF SPECIES. white, and on potato yellowish- white. In broth they produce turbidity and a scum on the surface. They reduce nitrates. They occur in sewage. Bacillus coli communis (Escherich).— See p. 344. Bacillus coli similis (Stern- berg). — Rods 1 to 3 /A in length, -4 to •5 p. in width ; singly and inpairs. Colonies circular and pale brown in colour. In the depth of gelatine they form a scanty growth in the track of the needle, and on the free sur- face a translucent film with irregu- lar margins. On potato the growth is pale brown or dirty white. They were isolated from human diver after death. Bacillus constrictus (Zimmer- mann). — Rods from 1'5 to 6'5 p. in length, and -75 /u, in width. The rods are segmented. Colonies are circular, granular, and greyish-yellow. In the depth of gelatine they form a filament in the track of the needle and irregular yellow heaps on the free surface. On the surface of agar the growth consists of a yellow shinin^layer, and on potato the same colour is produced. They occur in water. Bacillus coprogenes foetidus (Schottelius). — Rods about as large as Bacillus subtilis, but shorter. They are non-motile. Spore-for- mation occurs when the bacilli have access to the air, but not in the -animal body. In the depth of gelatine a fila- ment forms composed of yellow- ish compact colonies ; and on the surface a fine transparent film ; cultures emit a strong putrefactive odour. On potato they form a light grey, dry layer. Subcutaneous injection of small •doses had no effect on mice and rabbits, but very large quantities produced a toxic effect in rabbits. Swine are not affected. They were found by Schottelius in the intestine in cases of swine erysipelas. Bacillus coprogenes parvus (Bienstock). — Very short rods. On the surface of gelatine they form a very limited, almost in- visible, growth in the track of the needle. In mice they produce oedema and J death in thirty-six hours, and in | rabbits a local rash and death in eight days. They were isolated from human evacuations. Bacillus crassus aromaticus (Taratoff). — Rods 3'5 to 5 p long, 1-5 \i in width ; constricted in the centre. Colonies appear in the form of cup-shaped depressions and pro- duce a fruit-like odour. In gelatine they grow in the track of the needle, and later produce a funnel-shaped area of liquefaction. They occur in well water. Bacillus crassus sputigenus (Kreibohm). — Short thick rods, sometimes curved; capsulated. Colonies greyish- white. Cultures in gelatine resemble those of Fried- lander's pneumococcus. They are pathogenic in small animals. They were isolated from human sputum. Bacillus cuniculicida, Bacillus of Rabbit Styticcemia (see p. 228). Bacillus cuticularis (Tils).— Rods from 2 to 3 p. in length, '3 to "5 ft in width, and filaments. Colonies are yellow and the gela- tine is liquefied. In the depth of gelatine they produce liquefaction, and a skin forms on the surface. On potato the growth is slimy and yellow. They occur in water. Bacillus cuticularis a 1 b u s (Taratoff).— Rods 3'2 p long, con- stricted in the middle. Actively motile. Colonies are opalescent and bluish-white. Inoculated in the depth of gela- tine they form a white rosette- shaped growth on the surface and DESCRIPTION OF SPECIES. 507 a shining white growth along the needle track which sends off long rounded processes. On agar, glycerine agar and blood serum they produce a luxuriant white shining growth. Broth becomes turbid with a whitish deposit and pellicle. On potato there is a thick moist brown growth. They are found in water. Bacillus cyaneo-fuscus ( Bey erinck).— Rods '2 to -6 JJL in length, and half their length in width. Motile. In the depth of gelatine they produce colonies in the track of the needle, which later develop a black pigment. In broth with ^ per cent, of peptone they produce a blue colour, changing to brown and finally black. They were isolated from cheese, size and glue. Bacillus cyaneo-phosphores- cens (Katz).— Rods 2'6 p. in length, 1 n in width, singly, in pairs, and .filaments. Colonies circular and brownish or greyish-yellow. Inoculated in the depth of gela- tine they form a grey-white fila- ment in the track of the needle and liquefaction follows at the upper part ; later a skin forms on the sur- face and a yellow deposit occurs at the bottom of the liquefied jelly which has a reddish-brown tinge. In broth a similar skin floats on the surface and the broth is turbid. On sterilised fish the growth is viscid and yellow. Cultures are phosphorescent, especially in media containing excess of common salt. They were isolated from sea- water at Sydney, and are possibly identical with Bacillus phosphores- cens of Fischer. Bacillus cyanogenus (Hueppe). — Bacterium syncijiinum. liari/l//* •of Blue Milk.— Motile rods, 2'5 to c . •••»,«?•* \\ /\, ;\ FIG. 200. — BACILLUS CYANOGENUS, x 650. A. Ac- tive rods. B Rods in zooglpea. C. Chain of short rods. D. Chain of cocci. E. Cocci stage. F, G. Spore-forming rods. H. Involution- forms (NEELSEN). 3-5 n in length, and -4 n wide (Fig. 200). The rods after division may remain linked together, and form chains. Non-motile rods occur enveloped in a gelatinous capsule, and involution forms. Colonies appear after two days as small greyish-white points which gradually assume a moist appear- ance. The gelatine becomes steel- grey, throwing the white colonies into strong relief. In the depth of gelatine a whitish growth appears in the track of the needle near the upper part, and on the free surface, producing also a dark steel-blue discoloration of the jelly which spreads down- wards. On agar a greyish growth ap- pears, and the agar is coloured dark brown. On potato a yellowish moist growth develops, the potato around it is stained grey-blue. Milk be- comes slightly alkaline and of a slate-grey colour, which on the addition of acid changes to an intense blue. Milk in which the 508 DESCRIPTION OF SPECIES. lactic acid bacillus is growing be- comes sky-blue from the first. They are non-pathogenic. They are present in blue milk. Bacillus cyanogenus (Jordan). — Rods 1-3 p. in length, '8 p. in width. Slightly motile. Colonies granular and irregular. They colour the surrounding gela- tine brown. Inoculated in the depth of gela- tine the bacilli produce a scanty growth in the track of the needle, and a film on the surface with coloration of the gelatine beneath it. On agar they form a white layer, and the jelly is coloured brown. On potato the growth is brown. They were isolated from sewage. Bacillus cystiformis (Clado). — Short slender rods. Motile. Colonies circular, yellowish, gra- nular. Inoculated in the depth of gela- tine there is a scanty growth in the track of the needle, and a white patch on the free surface. On agar the growth is yellowish- white. They were isolated from urine. Bacillus delicatulus (Jordan), r— Rods 2 p, long and 1 p, broad, often in pairs or short chains. Actively motile. Spore-formation not observed. Colonies at first whitish with a radiating edge. Later they liquefy the gelatine and the centres become dark. Inoculated in the depth of gela- tine they rapidly liquefy it and form a whitish pellicle and a brown deposit. On agar a greyish crinkled growth appears, which gradually becomes white and shining. On potato there is a grey flat growth. Milk is coagulated and becomes strongly acid. Broth is made turbid and a white serum and precipitate formed. They occur in sewage. Bacillus dentalis viridans (Miller).— Rods slightly bent, singly and in pairs. Colonies circular, yellowish and concentric. Inoculated in the depth of gela- tine they grow in the track of the needle and on the free surface, and the jelly is coloured green. On agar the growth is colourless or slightly grey. Intraperitoneal injections in mice and guinea-pigs produce a fatal re- sult. Subcutaneous injections cause suppuration. They were isolated from caries of the teeth. Bacillus dentriticus (Bordoni Uffreduzzi and Lustig). — Rods "85 to 2-8 p. long, and '5 to -85 p. broad, singly and in zoogloea. Motile. Spore-formation not observed. Colonies have an arborescent appearance. Inoculated in the depth of gela- tine they form a circular raised growth at the point of puncture, and white colonies along the needle track. The jelly is gradually liquefied. On agar and blood serum there is a scanty growth on the surface and an abundant growth in the track of the needle. Blood serum is liquefied after some time. Broth is rendered turbid ; a white firm pellicle forms. On potato there is a thick moist white growth, which later becomes yellow. Found in water. Bacillus devorans (Zimmer- mann). — Rods -99 p, to 1*2 p. in length, '74 p. in width ; singly, in pairs and in chains. Colonies are circular, granular, and grey, with periphery formed of radiating processes. In the depth of gelatine they produce a whitish filament and an excavation at the upper part, which may or may not contain liquefied On agar a greyish film is found. They do not grow on potato. They occur in water. Bacillus diffusus (Frankland). — Rods 1/7 p. in length, '5 p. in width ; singly, in pairs, and fila- ments. DESCRIPTION OF SPECIES. 509 Colonies circular, bluish-green, •with a granular nucleus and delicate irregular periphery. In the depth of gelatine there is scarcely any growth in the track of the needle, but a shining greenish- yellow film on the surface, and liquefaction below it. On agar the growth is faintly yellow. In broth they produce turbidity and a yellowish deposit. On potato the growth is yellow- ish. They occur in earth. Bacillus diphtheriae (p. 332). Bacillus diphtheriae colum- barum (p. 336). Bacillus dysodes(Zopf).— Cocci, long and short rods, and spores. They were observed in bread, making it greasy and unfit for food, and generating a penetrating odour resembling a mixture of pepper- mint and turpentine. A great loss may result to bakers if the fungus is introduced with the yeast. Bacillus endocarditidis cap- sulatus (Weichselbaum).— Cocci resembling Fried lander's pneumo- cocci. Colonies faintly yellow, with dentated contours. In the depth of gelatine the growth produces a filament in the track of the needle, and a patch like stearin on the free surface. Large doses injected subcutane- ously, or into the peritoneal cavity prove fatal to rabbits. They were isolated from infarcts in a fatal case of endocarditis. Bacillus endocarditidis griseus. —Rods motile. Colonies granular and brown or yellowish-brown. In the depth of gelatine there is a filamentous growth in the track of the needle, and a circular whit- ish patch on the surface. On agar and potato the growth is greyish- brown. Cultures cause a fatal result in mice and guinea-pigs. They were isolated from a case of endocarditis. Bacillus enteritidis (p. 372). Bacillus epidermidis (Bordoni Uffreduzzi).— Rods 2-8 to 3 /i in length, and -3 \i in breadth. Spore- formation occurs at 25° C. They grow very sparingly on gelatine. On agar there is a surface growth. On potato at 15° to 20° C. the growth appears first in the form of drops, which gradually extend and coalesce and form a thin layer i-NN . ! FIG. 201. — PURE CULTIVATION OF BA- CILLUS FHJUKAXS ON THE SURFACE OF NUTRIENT AGAR-AGAR. I over the surface. On blood serum they form a thin film. Inoculation in rabbits and guinea- i pigs and on the human skin pro- | duced no result. They were isolated from flakes of cuticle from between the toes. Bacillus erysipelatis suis (p. 356). Bacillus erythrosporus i (Eidam). — Slender rods and 510 DESCRIPTION OF SPECIES. filaments. Motile, tion present. Spore-forma- I On potato the growth is brown. They occur in water. FIG. 202.— PHOTOGRAPH OF PART OF AN IMPRESSION PREPARATION OF BACILLUS FIGURANS ON NUTRIENT GELATINE, x 50. Colonies circular, with brown nu- cleus and yellowish-green periphery. In the depth of gelatine they Bacillus figurans (Crookshank). — Rods, with rounded ends, varying in length. Spore-formation present. FIG. 203.— PART OF THE SAME SPECIMEN SHOWN IN FIG. 202 x 200. grow in the track of the needle and on the free surface, colouring the jelly green by transmitted light, yellow by reflected light. In plate-cultivations they cause a cloudy growth, spreading from various points ; if a cover-glass impression is made, this is found DESCRIFFION OF SPECIES. 511 to consist of the regularly-arranged parallel rods. The chains of rods become twisted at intervals into curious convolutions, from which offshoots are continued in various directions. These long shoots or processes are again twisted at intervals into varying shapes and patterns (Figs. 202, 203). Culti- vated in nutrient gelatine, the bacilli form on the surface visible windings, from which fine filaments grow down into the gelatine. They spread out also in almost parallel lines transversely from the needle track. The gelatine is not liquefied. On an oblique surface of nutrient agar-agar the filaments spread down- wards into the substance of the jelly, and outwards from the cen- tral streak on the surface, forming a feather-like cultivation (Fig. 201). They were obtained from the air, and later were identified by the author with Bacterium Zopfii. Bacillus figurans (Vaughan).— Rods and threads. Colonies composed of curved and terlacing lines. In gelatine they grow in the track of the needle, and very slowly liquefy it. On agar they form a thin white layer. They were found in water. Bacillus filiformis (Tils).— Rods 4 p. in length and 1 /z in width, singly and in chains. Spore-forma- tion present. Colonies are granular, with yel- lowish nucleus. Inoculated in the depth of gela- tine there is no growth in the track of the needle, but a whitish growth on the surface, liquefying the jelly slowly. In broth they form a skin on the surface. On agar the growth is white. On potato the growth is dry and after a time brownish. They coagulate milk. They occur in water. Bacillus filiformis Havaniensis (Sternberg). — Long slender rods, 3 /A in diam., and filaments. Colonies circular, irregular ; deep colonies are brownish ; superficial colonies thin and translucent. Inoculated in the depth of gela- tine the growth is scanty in the track of the needle. In the depth of agar an opaque branching growth occurs in the track of the needle, and a scanty milk-white growth on the surface. In broth they cause opalescence. They were isolated from the liver in fatal cases of yellow fever. Bacillus flavescens (Pohl).— Rods 2-1 to 2-2 fj, in length, -8 /A in width. Slightly motile. Colonies yellow and granular. Inoculated in the depth of gela- tine the bacilli produce a filament in the track of the needle, and a growth spreads over the free sur- face. On the surface of agar the growth is composed of isolated yellow colonies. On potato they grow rapidly, forming a shiny yellow layer. They occur in marsh water. Bacillus flavocoriaceus (Ada- metz). — Minute rods occurring in zoogloea. Colonies circular, sulphur-yellow. Under a low power they show a brownish-yellow nucleus and yellow periphery. Inoculated in the depth of gela- tine the growth is granular in the track of the needle and on the free surface. They occur in water. Bacillus flue-res cens aureus (Zimmermann). — Short rods, 1/9 \L in length, '74 p. in width. Singly, in pairs and in masses. Motile ; flagellated. Coloniescircular, granular, yellow. Inoculated in the depth of gela- tine they form a filament in the track of the needle and a yellow patch on the surface. On agar the growth is golden- yellow, and the same on potato. They occur in water. Bacillus fluorescens lique- faciens (Flugge). — Short rods with rounded ends. Colonies on plates develop an iridescence around them. 512 DESCRIPTION OF SPECIES. In the depth of gelatine a white filament forms in the track of the needle, with liquefaction in the upper part, and an iridescent sheen is produced in the jelly. On potato they develop a brown- ish layer. They occur in water and in putrid infusions. Bacillus fluorescens lique- faciens minutissimus (Unna and Tommasoli). — Rods 1 '5 to 2 /u. in length, -3 p. in width. Colonies circular, with brownish nucleus and yellowish marginal zone. In the depth of gelatine lique- faction forms in the track of the needle. The liquid is turbid and yellowish, and a white sediment forms at the bottom and a fluores- cent scum on the surface. On agar and potato the growth is brownish. They were isolated from eczema- tous skin. Bacillus fluorescens longus {Zimmermann). — Rods varying in length, some curved, 1'45 to 1'65 p in length, '83 p, in width, and wavy filaments 14 p in length. Motile. Colonies circular, well-defined, yellowish, with broad twisted markings. On the surface of gelatine they produce a layer with a bluish-green fluorescent colour. On agar a thin film forms, and the jelly is coloured greenish- yellow. On potato the growth is slimy and yellowish. They occur in water. Bacillus fluorescens nivalis (Schmolck). — Rods and chains. Motile. Colonies circular, surrounded by liquid fluorescent jelly. Inoculated in the depth of gela- tine they produce liquefaction in the track of the needle, and the jelly is coloured green. On agar the growth is white and the jelly green. On potato the growth is brownish. They are probably identical with Bacillus fluorescens liquefaciens. They were isolated from snow. Bacillus fluorescens non-lique- faciens (Eisenberg). — Short deli- cate rods. Non-motile. Colonies have the appearance of fern-leaves, and are opales- cent. Inoculated in the depth of gela- tine the growth is very slight in the track of the needle, and on the surface filmy and fluorescent. On agar they form a greenish layer. They produce a brownish layer on potato, and bluish-grey dis- coloration. They occur in water. Bacillus fluorescens putidus (Fltigge). — Short rods with rounded ends. Motile ; spore-formation not known. They form small dark colonies with a greenish sheen, which have a penetrating odour. Inoculated in the depth of gela- tine they produce a pale-grey growth, and after three days colour the medium with a greenish tinge spreading down from above. On potato they rapidly develop a brownish layer. They occur on decomposing sub- stances, producing a greenish coloration. According to Lehmann and Neu- mann the Bacillus fluorescens albus and Bacillus fluorescens longus of Zimmermann and Bacillus fluores- cens non-liquefaciens are merely varieties of Bacillus fluorescens putidus ; and further, cultures of this bacillus on agar and on potato and in milk and in broth cannot be distinguished from those of Bacillus fluorescens liquefaciens. Bacillus fluorescens tenuis (Zimmermann). — Rods 1 to 1'85 fj. in length, '8 p. in width. Singly, in masses, and filaments. Motile. Colonies irregular. Inoculated in the depth of gela- tine a delicate filament forms in the track of the needle, and on the surface a greyish-white growth spreads and colours the gelatine yellow. On agar the growth is shining DESCRIPTION OF SPECIES. 513 and greenish, and on potato yellow- ish. They occur in water. Bacillus foetidus (Bacterium fcetidum, Thin). — Cocci, short rods, long rods, and leptothrix. The cocci, 1'25 to 1-4 /A in diam., occur singly or in pairs. Spore-formation present in the rods. They were isolated from the exudation in a case of profuse sweating of the feet, and the odour was noticeable in the cultivation Pif the stomach. Bacillus in cholera in ducks (Cornil and Toupet), p. 230. Bacillus in choleraic diarrhoea (Bovet). — Rods 2 to 4 p in length, 1 to 1*5 p in width, singly and in pairs, and filaments. In the depth of gelatine a filament forms in the track of the needle and a greyish transparent layer on the surface. On agar a greyish film is formed. On potato the growth is yellowish and abundant. Intra-peritoneal injections in guinea-pigs cause peritonitis and death. They were isolated from a case of choleraic diarrhoea. Bacillus in diphtheritic disease of Calves (Bacillus vitulonnit Loftier). — Rods about five or six times as long as wide, mostly united in long threads. A piece of membrane from a diph- theritic disease in a calf, placed on blood serum developed a white layer composed of the bacteria. Succes- sive generations were not obtainable. Mice inoculated directly from the calf died of a characteristic illness, and the same long bacteria were again found in the inoculated animals accompanying widespread infiltration, starting from the point I of inoculation. Inoculation of DESCRIPTION OF SPECIES. 517 guinea-pigs and rabbits gave doubt- ful results. They were found in the deeper stratum of pseudo- diphtheritic patches in calves. Bacillus in disease of bees (p. 471). Bacillus in erythema node-sum (Demme). — Rods 2'2 to 2*5 /x in length, '5 to '1 p. in width. They can be cultivated at 37°C. Colonies on agar are white with radiating lines. The bacilli inoculated in the depth of agar grow in the track of the needle, and produce peculiar off- shoots in the surrounding jelly. They are said to produce an eruption resembling erythema nodosum when inoculated subcu- taneously in guinea-pigs. They were obtained from the eruption and the blood in cases of erythema nodosum. Bacillus in fowl enteritis (Klein), p. 230. Bacillus in gangrene (Tricomi). — Rods 3 ft in length, I p, in width, singly and in pairs. Colonies circular, granular, dirty- yellow. In the depth of gelatine they produce a filament composed of closely aggregated colonies, and at the upper part conical liquefaction of the jelly, beneath a cup- shaped excavation. On agar and potato the growth is white. In jectedsubcutaneously in rabbits and guinea-pigs they produce gan- grene and death in a few days. They were isolated from a case of senile gangrene. Bacillus in grouse disease (Klein), p. 230. Bacillus in hog cholera (p. 351). Bacillus in infantile diarrhoea (Booker). — Rods morphologically identical with Bacillus coli com- munis. There are seven varieties of this bacillus. They were iso- lated from cases of infantile diar- rhoea. Bacillus in infantile diarrhoea (Lesage). — Rods 2 '4 p. in length, '75 ft in width, and filaments. Colonies irregular ia contour, colouring the gelatine green. On the surface of agar they form a greenish growth, and the gelatine is coloured green. Injected intravenously in a rabbit they produced diarrhoea. They are said to be identical with Bacillus fluorescens liquefaciens Bacillus in intestinal diph- theria in rabbits (Ribbert).— Rods 3 to 4 ft in length, 1 to 14 ft in width ; singly, in pairs, and in filaments. Colonies greyish ; granular. They produce in gelatine a deli- cate growth in the track of the needle. They are pathogenic. They were isolated from the intestine of rabbits suffering from a diphtheritic inflammation of the mucous membrane. Bacillus in jequirity infusion (see Bacillus ofSatOer). Bacillus in'measles (p. 283). Bacillus in noma (Schimmel- busch). — Rods singly, in pairs, and filaments. Colonies circular, greyish-white, granular, with irregular margins. In the depth of gelatine they produce a granular filament anda patch on the surface. On agar and potato the growth is greyish-white. They are pyogenic in rabbits. They were inoculated from a case of noma. Bacillus in ophthalmia (p. 190). Bacillus in potato rot.— Rods 2-5 to 4 /A in length, '7 to *8 ft in width ; singly, in chains, and in fila- ments. Spore-formation present. The bacilli inoculated in the depth of gelatine produce a funnel- shaped area of liquefaction. On agar the growth is composed of greyish-white slimy colonies. They were isolated from diseased potatoes. Bacillus in purpura haemor- rhagica (Tizzoni ana Giovannini). —Rods -75 to 1-3 ^ in length, "2 to •4 ft in width, singly, in pairs, and in masses. Colonies have a greyish-yellow nucleus and a marginal zone of fine 518 DESCRIPTION OF SPECIES. filaments both in gelatine and agar. Cultures produce a disagreeable odour. Subcutaneous in j ections in guinea- pigs and rabbits produce local oedema and death, with hsemorrhages in the internal organs. They were isolated from the blood in fatal cases of purpura in children. Bacillus in putrid bronchitis (Lumnitzer). — Rods 1-5 to 2 p. in length, slightly curved. They can be cultivated at 37° C. The colonies on agar are greyish- white. The bacilli inoculated on blood serum produce colonies which co- alesce and form a greyish-white film. Cultures have a disagreeable odour. Injected into the lungs of rabbits they produce purulent inflammation. They were isolated from the sputum in cases of putrid bronchitis. Bacillus in "red-cod" (Dautec). — Rods similar to Bacillus tetani, with terminal spore-formation. Colonies are circular ; reddish. On the surface of obliquely solidified gelatine they form a red growth in the track of the needle slowly followed by liquefaction. Cultivated on dried cod they produce a red colour. They were isolated from red-cod. Bacillus in rhinoscleroma (p. 411). Bacillus in saliva (Fiocca).— Very short rods -2 to -33 /x in width. Colonies circular, granular, and yellowish. On the surface of obliquely solidified gelatine they form a growth composed of transparent droplets. On potato they form a transparent film. In broth flocculi appear. They are pathogenic in rabbits and other small animals, and they are probably a variety of the bacillus of haemorrhagic septicaemia. They were isolated from saliva of cats and dogs. Bacillus in whooping cough (Afanassiew). — Rods -(j to 2-2 p in length, singly, in pairs, and short chains. Colonies granular, brownish. Inoculated in the depth of gela- tine there is a scanty growth in the track of the needle and a greyish growth on the free surface. On agar the growth is greyish. On potato the growth is shining and yellowish or brownish. They are said to produce symp- toms in rabbits and dogs compar- able to those of whooping cough. They were isolated from the throat in cases of whooping cough. Bacillus incanus (Pohl).— Rods 1*2 IJL in length, "8 p, in width. They produce rapid liquefaction in the track of the needle when inoculated in the depth of gelatine. On agar a thick white growth develops. On potato a whitish growth spreads over the surface. They were isolated from the water of marshes. Bacillus indicus (Koch).— Very short rods with rounded ends. FIG. 204.— BACILLUS INDICUS COLONIES IN NUTRIENT AGAR, x 60. The colonies have a scarlet tint. They are round, ovoid, or spindle shaped, and have granular margins. In the track of the needle beneath the surface no pigment is formed. Cultivated in nutrient gelatine they liquefy it and colour it crimson, and the growth of a darker crimson hue subsides to the bottom of the tube. On the surface of nutrient agar- agar the appearances are very characteristic. In a pure cultiva- tion a brilliant vermilion-coloured reticulated pellicle develops on the surface. (Plate II. Fig. 3.) DESCRIPTION OF SPECIES. 519 They form a vermilion layer on potato. They were isolated by Koch in India from the intestinal contents of an ape. Bacillus indigogenes (Alvarez). Rods 3 p in length and 1'5 /u in width, singly, and in chains ; capsu- lated. On agar they produce a yellowish- white layer, and are said to develop an indigo-blue colour in infusions of leaves of the indigo plant. Intravenous injections in guinea- pigs are said to produce death in a few hours. They were isolated from the leaves of the indigo plant. Bacillus indigonaceus (Clas- sen).—Rods 1-6 to 3 p. long, -8 to •9 p. wide ; non-motile. They form a sky-blue layer on the surface of gelatine. On potato the growth is dark- blue, and later has a metallic lustre. Bacillus indigoferus, which was found in water at Kiel, is only to be distinguished by its motility. Bacillus inflatus (A. Koch).— Rods 4'6 to 5'5 /A in length, '6 to 1-8 fi in width, and filaments. The colonies send out delicate processes. The bacilli inoculated in the depth of gelatine send out fine filaments in the track of the needle followed by slow liquefaction. On agar they form a shining brownish layer. In broth a pellicle forms on the surface. They occur in the air. Bacillus inunctus (Pohl).— Rods 3'5 n in length, -8 to *9 /x in width. Inoculated in the depth of gela- tine they grow both in the track of the needle and on the surface ; liquefaction follows in time. On agar they form a whitish growth. They were isolated from the water of marshes. Bacillus invisibilis (Vaughan). —Rods ; motile. The colonies are irregular and yellowish. Inoculated in the depth of gela- tine they grow both in the track of the needle and on the surface. On agar they form a white growth. On potato they develop an in- visible layer. They occur in water. Bacillus iridescens (Tataroff).— Rods from 3'5 to 5'2 p in length and threads. Spore-formation pre- sent ; slightly motile. The colonies have a characteristic appearance recalling that of the convolutions of the brain. Inoculated in the depth of gela- tine there is a depression at the point of puncture, and a thread- like growth along the needle track. On agar they form a thick, un- even, moist, greenish-yellow, irides- cent growth, with a pitted surface. Blood serum is liquefied. On potato there is a dry, thick, dark yellow growth like honey. Broth is rendered turbid, and there is a yellow deposit. They are found in water. Bacillus lactis aerogenes (Es- cherich). — Rods short and thick, •5 to -8 p broad, and 1 to 2 p long, with rounded ends ; usually in pairs side by side and also in irregular heaps. Non-motile : spore-forma- tion not observed. They grow best at 37°C. Colonies on the surface are raised, moist, shining and porcelain-white. Below the surface they have a yellowish nucleus. Inoculated in the depth of gela- tine the rods form an abundant nail-shaped growth. On potato the culture is composed of white colo- nies, and bubbles are formed. The colonies may coalesce and produce a creamy layer. On blood serum there is a raised, moist, shining, white growth. In milk sugar or grape sugar solutions they produce gas. Injected subcutaneously in rab- bits and guinea-pigs they cause death in from one to three days, and the bacilli are found in the blood and internal organs. 520 DESCRIPTION OF SPECIES. They were found in the intestinal tract of animals fed with milk and of infants at the breast. Bacillus lactis albus (Loffler). — Rods, 3-4 p. in length, '96 /* in width, and filaments. Spore-forma- tion present. Inoculated in the depth of gela- tine they slowly liquefy the upper part, and a white scum forms on the surface. On agar they form a white layer. On potato the growth is dry and white. They coagulate milk. They occur in milk. Bacillus lactis erythrogenes (Hueppe). — Short rods, 1 to 1/4 p, in length and *3 to -5 p in width, and filaments. Colonies small and circular ; greyish- white ; later yel- low and surrounded by liquefied gelatine with a pink tinge. In the depth of gelatine the growth in the track of the needle is scanty, but on the surface a whitish patch forms which after- wards turns yellow, and the gela- tine is coloured pink. Later lique- faction sets in, and the liquefied gelatine is turbid and pink. On agar a shining yellow layer develops, and the same on potato. In broth the bacilli produce tur- bidity, and they coagulate milk. They occur in " red milk." Bacillus lactis pituitosi (Ldf- er). — Rods slightly bent. Colonies circular, greyish-white. On agar and potato they pro- duce a greyish- white layer. They render milk viscid. They occur in milk. Bacillus latericeus (Adametz and Eisenberg). — Rods and fila- ments. Colonies circular, granular, red- dish-brown. In the depth of gelatine there is a scanty growth along the track of the needle and a brick- red growth on the surface. On potato the growth is also brick-red. They occur in water. Bacillus leporis lethalis (Gibier and Sternberg). — Rods 1 to 3 p. in length, -5 /u. in width. Colonies transparent and with the appearance of broken glass. In the depth of gelatine there is a growth along the track of the needle with a conical area of lique- faction at the upper part, and a white sediment. On agar they form a trans- lucent film. They liquefy blood serum. On potato the growth is pale- yellow. Cultures injected into the peri- toneal cavity of rabbits are toxic. They were isolated from the in- testinal contents in cases of yellow fever. Bacillus leprae (p. 407). Bacillus leptosporus (L. Klein). — Rods resembling hay-bacilli, singly, in chains and long twisted filaments. The spore-membrane is said to form part of the newly grown bacillus, and the filaments are de- scribed as possessing peculiar spas- modic movements. They were isolated from a con- taminated culture. Bacillus limbatus acidi lactici (Marpmann).— Rods short, thick : singly, in pairs ; capsulated. Colonies white. In the depth of gelatine they develop slightly in the track of the needle, and produce a white patch on the free surface. In milk they produce coagula- tion and lactic acid. They occur in milk. Bacillus limosus (Russell).— Rods 3 to 4 p, in length, 1/25 \i in width ; singly, in pairs and chains ; spore-formation present. Colonies transparent, surrounded by filamentous processes. In the depth of gelatine pre- pared with sea-water, liquefaction occurs rapidly in the track of the needle, and a deposit forms at the bottom and a thin skin on the surface. On agar they form a white layer, and in broth turbidity and a thick scum. On potato the growth is greyish- white. DESCRIPTION OF SPECIES. 521 They were isolated from deep- sea dredgings. Bacillus liodermos (Fliigge).— Small short rods with rounded ends ; actively motile. Colonies with irregular outlines float on liquefied gelatine in the form of small white flakes. Inoculated in gelatine a greyish growth occurs along the track of the needle, but the medium later be- comes liquefied and a greyish-white flocculent deposit settles at the bottom. On potato a smooth shining yel- lowish-white layer spreads quickly over the whole surface, and after some days becomes opaque and slightly wrinkled. They occur on potato. Bacillus liquefaciens (Eisen- berg). — Rods short and thick, with rounded ends. Very motile. Colonies round, with smooth edges and slimy centres. Lique- faction follows, and a putrefactive odour is noticed. In gelatine they make a funnel- shaped whitish growth along the track of the needle. On potato the growth is pale yellow. They occur in water. Bacillus liquefaciens commu- nis (Sternberg).— Rods 1 to 2 p in length, and •? //. in width ; singly and in pairs. In the depth of gelatine they produce rapid liquefaction in the track of the needle. On potato a wrinkled pinkish layer is formed. They were isolated from the eva- cuations of yellow-fever patients. Bacillus liquefaciens magnus (Liideritz). — Rods 3 to 0 p in length, -8 to 1-1 p. in width, and filaments. They are anaerobic. Colonies develop below the sur- face of the gelatine, and liquefaction extends upwards to the surface. The bacilli inoculated in the depth of gelatine cause liquefaction in the lower part of the track of the needle. In the depth of agar the colonies have delicate branches. They liquefy blood-serum, and produce a putrefactive odour. They occur in earth. Bacillus liquefaciens parvus (Liideritz). — - Rods 2 to 5 n in length, -5 to •? M in width, and filaments. They are anaerobic. Colonies are white, and liquefy gelatine ; but in agar they are spherical or almond-shaped. In the depth of gelatine isolated colonies appear in the track of the needle, and in the depth of agar there is gas formation. They occur in earth. Bacillus liquidus (Frankland). — Rods short and flat with rounded ends, usually in pairs, the length of each pair varying from T5 pi to 3*5 p. They are very variable in size ; highly motile ; spore-forma- tion not observed. Colonies form cup-shaped excava- tions, with almost clear, colourless contents. The edges are at first smooth and circular, but they become serrated and granular, and soon coalesce. A broad funnel-shaped depression forms along the whole track of the needle, containing turbid liquid and masses of flocculent material. Later a thin pellicle forms on the surface, which sinks if the tube is shaken. On agar they grow quickly, form- ing a smooth shining layer. On potato a thick flesh-coloured growth appears. Broth is rendered turbid with an abundant sediment, and after a few days a pellicle forms. They are common in unfiltered water. Bacillus litoralis (Russell).— Colonies granular, with regular contour ; slowly liquefying. In the depth of gelatine they develop a growth in the track of the needle, and at the upper part produce a cup-shaped cavity lined with the culture. The gelatine is tinged with brown in the vicinity. On agar they produce a greyish- white film, and in broth turbidity. Inoculated in the depth of gelatine the bacilli form a funnel-shaped liquefaction along 522 DESCRIPTION OF SPECIES. the track of the needle, and the whole of the gelatine gradually becomes liquid, with a flocculent deposit at the bottom and a greyish wrinkled stain on the surface. On potato a thick wrinkled whitish skin forms, which rapidly grows over the whole surface. On at- tempting to raise this skin it will be found to be attached to the potato by a mucous substance which may be drawn out in long threads. According to Hueppe the bacilli cannot form any ropy sub- stances from sugar, but they have an energetic diastatic action. They coagulate the casein in milk in a similar manner to rennet. The bacilli are ubiquitous. Bacillus lividus (Plagge and Proskauer) . — Rod s . Colonies blue-black, liquefying. In the depth of gelatine they produce a colourless thread in the track of the needle and a violet layer on the surface followed by gradual liquefaction. On agar the growth is blue-black, and on potato violet. They were isolated from water. They are probably identical with Bacillus ianthinus, or merely a variety. Bacillus luteus (Fliigge). — Short immotile rods. Colonies irregular in form, appear brownish under a low power, and yellow to the naked eye. In test-tube cultivations they form a yellow growth without liquefying the gelatine. They occur contaminating plate- cultivations. Bacillus maidis (Cuboni). - Rods 2 to 3 p. in length, singly, in pairs, and in chains ; spore- formation present. Colonies granular, with wrinkled periphery ; later, liquefying. In the depth of gelatine they produce rapid liquefaction in the track of the needle. On agar a dry wrinkled white film spreads over the surface. On potato the growth is wrinkled, and later yellowish-brown. They liquefy blood serum. They were isolated from human evacuations and infusions of maize. Bacillus mallei (p. 452). Bacillus megatherium (De Bary). — Rods 2 -5 p. wide and four to six times as long, with rounded ends and slightly curved, and in short irregular chains. Transverse division . occurs, each segment attaining the length of the original rod. In the fresh state they appear non-articulated, but when treated FIG. 205.— BACILLUS MEGATHERIUM. (a) A chain of rods x 250, the rest x 600. (b) Two active rods : d and /, successive stages of germination ; h and I, successive stages of germina- tion. (De Bary. ) with a dehydrating agent they are seen to be composed of short seg- ments with granular contents. They are motile. Colonies are small and circular, and the gelatine is liquefied. In the depth of gelatine the bacilli grow rapidly, forming a | funnel-shaped liquefaction in the ! upper part. On agar they form a whitish layer on the surface, and the jelly ' acquires a dark colour. On potato yellowish-white cheesy colonies are formed round the point I of inoculation. In cultures there I is copious spore-formation. They ; grow best at 20° C. They were isolated originally from boiled cabbage. DESCRIPTION OF SPECIES, 523 FIG. 206. — PURE-CULTURE OF BACILLUS MEGATHERIUM IN GELATINE. Bacillus membranaceus ame- thystinus (Eisenberg). — Short rods with rounded ends from 1 to 1*4 fj. long, and "5 to '8 \i broad. They are grouped together irregularly. Some individual bacilli stain more deeply at the ends than in the middle. Non-motile. They grow only between 15° and 20° C. Spore- formation uncertain. The colonies gradually assume a violet hue, and after liquefying the gelatine float on the surface as violet pellicles, resembling a membrane stained with gentian violet. Inoculated in the depth of gelatine a yellowish -white growth appears on the free surface, which after ten days or more becomes violet. Liquefaction takes place gradually, and in about a month a thick violet layer covers the gela- tine which lies beneath the liquid part. On agar the growth, which at first has a yellowish milky appearance, becomes violet after eight or ten days. In three or four weeks it has become very much wrinkled, and has a beautiful deep-violet colour with a metallic lustre. The jelly is not stained, and the growth can be easily removed from its surface. On potato they grow slowly, and form a dirty yellow or olive-green colour. In broth they grow very slowly ; after some weeks a violet deposit and pellicle are formed, and the liquid between becomes dark brown. They were found in well water. Bacillus meningitidis puru- lentae (Neumann and Schaffer). — Rods 2 /z in length, -6 to "1 p. in width, and filaments. Colonies granular, greyish. In the depth of gelatine a greyish- yellow filament develops, composed of closely packed colonies, and on the surface a greyish layer. On potato the growth is moist and white. They are pyogenic in small ani- mals and dogs. They were isolated from a case of purulent meningitis. Bacillus mesentericus fuscus (Fliigge). — Rods small and short, singly, in chains of two and four. Actively motile. Spore-formation present. Colonies are at first roundish and rather white, with a sharp outline : later delicate brownish-yellow pro- cesses appear. Liquefaction occurs rapidly- Inoculated in the depth of gela- tine a whitish growth forms along the track of the needle, the upper portion of which soon liquefies ; greyish flakes float in the liquefied portion. On potato a smooth yellowish growth appears on the first day, but it soon becomes brown and wrinkled. It remains relatively thin and superficial, and quickly spreads over the whole surface. They are found in hay dust, in the air, on the surface of potatoes, and are very widely distributed. Bacillus mesentericus ruber.— Slender rods, singly, in pairs, and in filaments. Colonies are circular and yellow- ish until they come to the surface, when they produce a network and 524 DESCRIPTION OF SPECIES. liquefy the gelatine. The network disappears and a little deposit occurs at the bottom of the liquefied area. Inoculated in the depth of gela- tine a whitish cloudy growth forms along the needle track, liquefaction sets in and extends until the gela- tine is completely liquefied. On potato a thin crinkled film is formed, which is yellowish or red- dish-yellow in colour. They occur on potato. Bacillus mesentericus vulga- tllS (Fliigge). — Rods large and thick, of ten forming pseudo-threads. They have an oscillating movement. Spore-formation present. The colonies are bluish-white and almost transparent, though the centres become gradually opaque. They sink in the liquefied gelatine, and are granular with irregular contour. Bacillus multipediculus (Fliigge). — Rods long and slender. Non-motile. The colonies consist of a central oval nucleus, from which numerous tapering processes shoot out mostly towards one pole. This form of growth gives a curious resemblance to an insect with feet and antennse. Inoculated in the depth of gela- tine a whitish line forms along the track of the .needle, from which short processes grow out. On potato a rather scanty dirty- yellow growth forms, and the sur- face of the potato becomes dis- coloured around it. They are often found as a con- tamination on potato. Bacillus muscoides (Liborius) — Rods 1 p, thick, sometimes form- ing threads ; slightly motile, and with round or oval spores at one end. They are anaerobic. The colonies ramify and resemble a delicate moss. They were found in the oedema- tous fluid of field mice inoculated with garden earth and stale cheese. Bacillus mycoides (Fliigge).— Rods rather thick, nearly the size of Bacillus arithracis. Motile, often forming long pseudo-threads. Oval and highly refractive spores both in the rods and threads. Colonies consist of a whitish tur- bidity in which colourless branched and interwoven processes are seen ; these increase rapidly, and after twelve to twenty hours appear like the mycelium of a fungus. Inoculated in the depth of gela- tine they form very fine and closely set hairs extending from the track of the needle. Later liquefaction occurs. On potato a whitish layer gradu- ally extends over the surface. They occur in earth from the surface of cultivated ground. Bacillus mycoides roseus (Scholl).— Rods. Colonies composed of interlacing filaments. Inoculated in the depth of gela- tine they produce liquefaction in the track of the needle ; a reddish scum forms on the surface, and a reddish deposit at the bottom of the liquefied area. On agar they produce, in the absence of light, a pink growth. Bacillus neapolitanus (Emme- rich).— Short rods '9 fj. in width. FIG. 207. — BACILLUS NEAPOLITANUS, x 700 (EMMEKICH). a, From intes- tinal contents in a case of cholera ; b, From peritoneal fluid of an inoculated guinea-pig. DESCRIPTION OF SPECIES. 525 Colonies circular, later irregular, granular, strongly refractive and yellowish - brown. They are probably identical with Bacillus coli communis. They were isolated from cases of cholera at Naples. Bacillus necrophorus (Loffler). — Rods and filaments. They cannot be cultivated on the ordinary media. In rabbit broth they give rise to fluify masses of filaments. Intravenous injection produced in rabbits a pysemic condition in about a week. The bacilli were found in the pus. They were isolated from a rabbit which had been inoculated with fragments of a condyloma. Bacillus nitrificans (Wino- gradsky). — Very small rods -5 /* in in length, singly and in zooglcea. Colonies in silica jelly are lenticular, and sub-cultures in liquid media produce a gelatinous de- posit. They are powerful oxidising agents. They were isolated from the soil. Bacillus nodosus parvus (Lustgarten). — Rods 1-2 to %2'4 p in length, -4 p in width ; singly and in pairs. Inoculated in the depth of agar they produce a white filament in the track of the needle composed of crowded colonies, and on the surface a hemispherical glistening growth. They were isolated from the human urethra. Bacillus nubilus(Frankland). — Slender rods 3 p. long and '3 /z wide, and threads. Single bacilli have an active rotatory movement, but the long threads in broth cultures are quite motionless. Spore-formation not observed. The colonies appear as cloudy undefined patches, which rapidly liquefy the gelatine. They consist of a tangled mass of threads. Inoculated in the depth of gela- tine they produce along the track of the needle horizontal circular plates, with a delicate cloud-like appearance, and liquefaction at the upper part. Later the whole of the gelatine is liquefied. On agar they form a thin opales- cent blue-violet film, the edges of which exhibit later a distinct violet fluorescence. On potato there is a slightly yellow growth which is scarcely visible. Broth is rendered turbid with a dirty-white deposit, the surface being covered by a thin pellicle. They occur in water. Bacillus ochraceus (Zimmer- mann). — Rods 1-25 to 4*5 \i in length ; -65 to '75 \L in width ; singly, in pairs, chains, and fila- ments ; capsulated. Colonies circular, granular, yel- low, liquefying. Inoculated in the depth of gela- tine they produce liquefaction in the track of the needle, and a. yellow deposit. On agar and potato the growth is yellow ochre in colour. They occur in water. Bacillus oedematis aerobicus (Klein).— Rods '8 to 2-4 /x in length, •7 fj. wide, and long filaments. Colonies greyish, transparent, with irregular contour. In the depth of gelatine a fila- ment occurs in the track of the needle, and gas bubbles in isolated colonies in its lower part, and a transparent patch with irregular margin on the free surface. On the surface of agar they pro- duce a greyish- white layer. In broth there is turbidity with I flocculi. On potato the growth is yellowish and viscid. They give rise to extensive oedema in guinea-pigv«i, and in a less marked ! form in rabbits. They occur in earth. Bacillus oedematis maligni ! (p. 2-20). Bacillus of Belfanti and Pas- car Ola.— Very short rods. Colonies circular, granular, yel- i lowish-grey. Inoculated in the depth of gela- tine they produce a filament com- ; posed of closely-packed minute 52(> DESCRIPTION OF SPECIES. colonies, and on the surface a greyish film. On agar they produce a greyish- white growth. On potato a transparent whitish film. They are fatal to rabbits, guinea- pigs and small birds. They are probably identical with Bacillus septicaemias haemorrhagicae. They were isolated from pus in a case of tetanus. They were isolated from deep-sea dredgings. Bacillus of Colomiatti.— Minute rods. Spore-formation occurs at the ends of the rods. They can be cultivated at 37° C. They form a thin film on agar and on blood serum. They were isolated in cases of conjunctivitis. Bacillus of Fulles, No. I.— Rods 1 to 1*2 p. in length, -6 /z in width. Colonies circular, granular, yel- lowish-brown. On the surface of gelatine they produce a thin film, and in broth turbidity and flocculi. On potato the growth is yel- lowish. No. II. Very short rods. Colonies circular, granular, yel- lowish. In the depth of gelatine the growth resembles Friedlander's pneumococcus. On potato the growth is yellowish. They were isolated from earth. Bacillus of Ghiillebeau.— No. I. Short rods 1 to 2 ft in length, 1 ^ in width. Colonies spherical, granular. In gelatine the bacilli produce a growth in the track of the needle and a white patch on the surface. On agar the growth is white, and on potato yellowish, viscid, and con- taining gas bubbles. They coagulate milk. No. II. Rods resembling the above described but distinguished by the production of viscid colonies and, extremely slowly, of liquefac- tion in the jelly. No. III. Rods also resembling the above mentioned, but colonies are adherent to the jelly and coarsely granular. Milk and other liquid culture media are rendered extremely vis- cid. Bacillus of Letzerich.— Rods sometimes bent, and filaments. They rapidly liquefy gelatine. They produce purulent peritonitis and death in rabbits. They were isolated from urine. Bacillus of Martinez (Stern- berg). — Short rods 1 to 1*2 ft in length and "5 to '8 /z in width ; non-motile. Colonies circular and translucent, with a central, nipple-like projec- tion, and the surface covered with mosaic markings. In the depth of gelatine the growth consists of large spherical translucent colonies in the track of the needle, and a thin, translucent, scanty growth upon the surface. They were isolated from the liver in a fatal case of yellow fever. Bacillus of Nocard. (Vide Strep, tothrix farcinica.) Bacillus of Okada.— Short rods rather thicker than the bacilli of mouse-septicasmia, singly, in pairs and in filaments. Spore-formation not observed. Colonies granular and brownish. Inoculated in the depth of gelatine they form a white filament, and on the surface a milk-white patch. Inoculated on agar the growth spreads over the surface forming a milk-white layer. In broth they produce cloudiness and a layer floating on the surface. They do not grow on potato. Cultures produce death in mice, guinea-pigs and rabbits in twenty hours. They were isolated from dust. Bacillus of Koth.— No. 1 and No. 2. Rods. Two varieties were isolated from old rags. They appear to be varieties of Bacillus coli communis. Bacillus of Sattler.— 2 to 4-5 /* long and '58 p thick. They can be cultivated on nutrient gelatine and blood serum. Infusion of jequirity containing DESCRIPTION OF SPECIES. 527 the bacilli, inoculated into the con- junctiva of healthy rabbits, produces severe ophthalmia. The poisonous principle is a chemical ferment dbrln. Boiling, which does not destroy the spores of the bacillus, destroys the ferment, and cultiva- tions started from these spores, though teeming with jequirity bacilli, are quite harmless (Klein). The bacilli occur in infusions of the beans of Abrus precatorius or jequirity. Bacillus of Schaffer (Freuden- reich). — Rods 2 to 3 /u in length, 1 fjt in width, and long filaments. Colonies circular, granular, yel- lowish. In the depth of gelatine a growth develops in the track of the needle and a greyish layer on the surface. On agar the growth is greyish and sometimes brownish, and on potato yellowish. In broth with peptone and milk sugar there is copious formation of gas-bubbles. They closely resemble Bacillus coli communis. They were isolated from cheese and potato. Bacillus of Scheurlen.— Rods T5 to 2*5 p, in length, '5 /* in width. They were isolated from cancerous growths by Scheurlen, and were later identified with Bacillus epider- midis. Bacillus of Schou. — Short rods and cocci-forms. Colonies are spherical, opaque, and granular. The bacilli inoculated in gelatine rapidly liquefy it, and a white de- posit forms at the bottom of the liquid. Rabbits inoculated in the trachea, or made to inhale pure-cultures, are said to develop fatal pneumonia. They were isolated from rabbits with pneumonia, following section of the vagi. Bacillus of swine plague (p. 351). Bacillus of Tommasoli.— Short rods from 1 to 1*8 /* in length, and •25 to '3 /x in width, singly, and in short chains. Colonies grey and shiny. In the depth of gelatine they form a filament composed of closely- packed colonies, and on the surface a shining mass. On agar the growth consists of greyish patches. On potato the growth is granular and yellowish-white. Cultures rubbed into the skin are said to produce a vesicular eruption. They were isolated from the scalp in a case of sycosis. Bacillus of Utpadel.— Rods 1-25 to 1'5 a in length, and "75 to 1 /u in width, singly, in pairs and short chains. Colonies milk-white. On the surface of gelatine the growth is milk-white, and on agar yellowish- white . Injected subcutaneously in cats, guinea-pigs and mice, they produce extensive oedema, and a fatal ter- mination. They were isolated from the human intestine. Bacillus of Winogradsky. (See Bacillus nitrificans.) Bacillus ovatus minutissi- mus (Unna). — Short rods with pointed ends '6 to '8 p in length, •4 ju. in width, singly, and in masses. Colonies are minute, granular and yellowish. The bacilli inoculated in the depth of gelatine form a filament of closely packed greyish-white colonies, and on the free surface there is a shiny, greyish- white layer. On agar the growth is very simi- lar, and on potato also. They were isolated from the skin in eczema seborrhceicum. Bacillus oxytocus perniciosus (Wyssokowitch). — Rods short and thick. Colonies circular, granular, yel- lowish, or yellowish-brown. The bacilli inoculated in the depth of gelatine produce a growth resembling Friedlander's pneumo- coccus. They coagulate milk. 528 DESCRIPTION OF SPECIES. The products injected intrave- nously produce death in from three to twenty-four hours. They occur in sour milk. Bacillus pestifer (Frankland). —Rods 2-3 p. in length, 1 ^ in width, and filaments. Motile. Colonies resemble those of Bacil- lus vermicularis. On agar they produce a dentated transparent layer, and on potato a flesh-coloured growth. They occur in the air. Bacillus phosphor escens geli- dus (Forster). — Very short rods. Colonies circular, granular, yel- lowish or greenish. The bacilli, inoculated in the depth of gelatine, produce very little . growth in the track of the needle, and a white film on the surface. On agar and potato the growth is whitish. Cultures are photogenic. They were isolated from phos- phorescent fish. Bacillus phosphorescens Indi- CUS (Fischer). — Rods singly and in pairs, and filaments. Motile. Colonies circular, well-defined, greenish. The bacilli, inoculated in the depth of gelatine, produce a greyish filament in the track of the needle, and a hemispherical excavation of the jelly at the upper part. Later the jelly is liquefied, and there is a yellowish scum on the surface. On agar and potato the growth is white. Cultures are photogenic. They were isolated from sea- water. Bacillus phosphorescens indi- genus (Fischer). — Rods 1-3 to 1'2 p. in length, €4 to '7 /* in width, singly, in pairs, and filaments. Colonies circular, greenish, and later yellowish. The bacilli, inoculated in the depth of gelatine, produce a conical excavation in the upper part of the needle track without liquid con- tents, but with a dry growth on the sides. There is no growth on potato. Cultures are photogenic. They occur in sea-water and on phosphorescent fish. Bacillus plicatus (Zimmer- mann). — Minute rods, singly, in pairs, and in short chains. Colonies yellowish-white. The bacilli, inoculated in the depth of gelatine, form minute isolated colonies, and on the surface a wrinkled patch with gradual lique- faction. On potato the growth is dry and yellowish. They occur in water. Bacillus pneumosepticus (Babes). — Short rods, '2 p in width. Colonies irregular, semi - trans- parent. In gelatine, the bacilli grow in the track of the needle. On agar the growth is whitish and shining. Rabbits, guinea-pigs and mice die in two or three days of septi- caemia when a culture is injected subcutaneously. They were isolated from a fatal case of septic pneumonia. Bacillus polypiformis (Libo- rius). — Slender rods, spore-forma- tion present. They are anaerobic. Colonies composed of peculiar convoluted processes. In the depth of blood serum they produce a cloudiness at the lower part of the needle track. They occur in soil. Bacillus prodigiosus (Micro- coccus prodigiosus : Cohn. — Blood rain, Bleeding host). Very short rods with rounded ends, and thread forms *5 to 1 /u, in width, forming at first rose-red and then blood-red zoogloea. They liquefy gelatine. They grow luxuriantly on the sloping surface of nutrient agar- agar, and on sterilised potato, and the colour varies from blood-red to bright-red with sometimes a metal- lic lustre. The cells themselves- are colourless. The colouring-matter resembles f uchsine ; it is insoluble in water but soluble in alcohol. The addition of acids changes it to carmine red, and of alkalies to a yellow colour. They appear occasionally on DESCRIPTION OF SPECIES. 529 bread, boiled rice, and starch paste, and more rarely on boiled white of egg and meat. Milk sometimes becomes coloured blood-red by the growth of this fungus, an appear- ance formerly attributed to a disease of the cow. In Paris in 1843 the micro- organism was peculiarly prevalent, attacking especially the bread pro- duced in the military bakehouses. Bacillus proteus fluorescens (Jager). — Short thick rods and threads. Actively motile. Colonies resemble minute drops of water. The rods inoculated in gelatine produce a growth similar to that of Koch's comma-bacilli. The jelly becomes greenish, and a pellicle forms on the surface. On agar the growth when fully developed is yellowish- white, with a green fluorescence. On potato they form a brown layer. They are pathogenic in mice. They were isolated from the internal organs of fowls suffering from an epidemic disease. Bacillus pseudo-diphtheriticus (p. 335). Bacillus pseudo-tuberculosis (Pf eiffer). — Rods varying in length. Colonies circular, with dark nu- cleus and transparent zone. In the depth of gelatine they produce a filament composed of small colonies, and on the surface a patch with concentric markings. They grow on agar, but not readily on potato. Inoculated in mice, guinea-pigs, rabbits, and hares, they produce a fatal result in from six to twenty days. An abscess forms locally, the lymphatic glands enlarge and caseate, and the internal organs contain nodules resembling tubercle. They were isolated from the in- ternal organs of a horse supposed to be glandered. Bacillus pulpae pyogenes.— Rods slightly bent and with pointed ends ; singly, in pairs, and in chains. Colonies circular, yellowish- brown. Inoculated in the depth of gelatine liquefaction occurs in the upper part of the needle track and ex- tends downwards. Intraperitoneal injection in mice produces death in from eighteen to thirty-six hours. They were isolated from putrid dental pulp. Bacillus punctatus (Zimmer- mann). — Rods 1 to 1-6 in length, •77 p. in width, singly, in pairs, and chains. Colonies composed of stringy masses in liquefied gelatine. The bacilli inoculated in the depth of gelatine produce rapid liquefaction in the track of the needle, and a white deposit. On agar the growth is smooth and shining. On potato the growth is brownish. They occur in water. Bacillus putrificus coli (Bien- stock). — Slender, motile rods, 3 //. in length, often less, sometimes FIG. 208. — BACILLUS PUTRIFICUS CoLir X 1000 (BlENSTOCK). forming long threads. Spore-for- mation present. Cultivations in gelatine are iri- descent. They are constantly present in faeces. Bacillus pyocyaneus (Gessard). — Slender rods, singly, in twos and threes, or in irregular masses. Spore-formation present. White colonies appear in twenty- four hours, which liquefy the gela- tine. The whole of the medium acquires a greenish shimmer. If the bacilli are cultivated in gelatine, the jelly is liquefied, and coloured green by reflected light, and a deep orange by transmitted light. On agar they form a white layer, and colour the medium a pea-green. 34 530 DESCRIPTION OF SPECIES. On potato a dry rust-brown growth appears at the seat of inoculation, which becomes green when treated with ammonia. The pigment formed by the micro- organism is a definite principle — pyocyanin. It can be extracted with chloroform from pus and from washing of bandages ; it is soluble in acidulated water, which it colours red. In neutral solution it becomes blue. It crystallises in chloroform in long needles ; and forms some- times lamellae and prisms. They cause death in guinea-pigs wh£h injected into the abdominal cavity. Eabbits are not killed by intravenous injection. The bacilli are antagonistic to anthrax bacilli. Charrin and others have shown that rabbits inoculated with a pure- culture of Bacillus pyocyanus after inoculation with Bacillus an thracis will not succumb to anthrax. Woodhead and Wood produced similar results by using sterilised cultures, showing that the results were due to the chemical products of the bacilli. The rods occur in the pus of those cases in which the wounds and pus- stained bandages exhibit a greenish- blue colour. Bacillus pyogenes feet id us (Passet). — Small rods, about 1'45 /u, in length, and -58 p. in width ; FIG. 209.— BACILLUS PYOGENES FCETIDUS, x 790 (PASSET). often in pairs, or linked together in chains. They are motile, and spore-formation occurs. Colonies like white points appear after twenty-four hours, and de- velop into greyish spots, and these •enlarging coalesce into a layer. Cultivated in nutrient gelatine a greyish, veil -like growth forms on the surface. In nutrient agar-agar the culti- vation resembles the growth in gelatine. On blood serum a moder- ately thick greyish-white streak develops, and on sterilised potato an abundant, shining, brownish culture. From all these media a putrid odour emanates, but no smell is detected from a cultivation in milk. Inoculated into mice and guinea- pigs, abscesses are produced or death from septicaemia results. They were isolated from putrid pus. Bacillus pyogenes soli(Bolton). — Rods resembling Bacillus diph- theriae. Colonies granular, faintly yellow. The bacilli inoculated in the depth of gelatine form colonies in the track of the needle. They are pyogenic in mice and rabbits. They are present in earth. Bacillus radiatus (Luderitz).— Rods 4 to 7 /i in length, *8 p in width, and filaments. Motile. They are anaerobic. Spore-forma- tion present. Colonies are composed of delicate interlacing filaments. In the depth of gelatine a growth occurs at the lower part of the needle track, from which fine fila- ments are given off in the sur- rounding gelatine, and liquefaction follows. The growth in the depth of agar is also composed of fine filaments. In sub-cultures they produce a cloudy liquefaction. Cultures have a peculiar odour. They occur in earth. Bacillus radiatus aquatilis (Zimmermann). — Rods 1 to 6'5 /* in length, -65 // in width. Colonies white, with a marginal zone of radiating filaments. The bacilli inoculated in the depth of gelatine grow in the track of the needle, and excavate and liquefy the surrounding gelatine, and form on the free surface a wrinkled patch, which later subsides DESCRIPTION OF SPECIES. 531 to the bottom of the liquefied area. On agar a smooth slightly brownish layer is formed, and on potato it is yellowish. They occur in water. Bacillus ramosus (Eisenberg).— Rods singly, and in chains ; fila- ments. Spore -formation present. Colonies are composed of curi- ously twisted filaments. The bacilli inoculated in the depth of gelatine produce delicate fila- ments extending in all directions from the track of the needle, fol- lowed by liquefaction ; later a skin forms on the surface. There is a sediment at the bottom of the tube. On agar they form a greyish filamentous layer, and on potato a whitish growth. They occur in earth and water. Bacillus reticularis (Jordan).— Rods 5 p, in length. 1 /x in width ; singly, and in short chains. Motile. Colonies are composed of radi- ating filaments, and liquefy the gelatine, forming an excavation with a reticulated lining. The bacilli inoculated in the depth of gelatine produce filaments extending from the track of the needle, and at the upper part the jelly is excavated in the form of a cup. On agar they form a dry layer, and on potato a white woolly growth. Broth is made turbid, and milk slowly coagulated. They occur in water. Bacillus rosaceus metalloides ( Bacterium rosaceum metalloides, Dowdeswell ; Magenta bacillus). — Rods -6 to -8 \i in breadth. Colonies in the depth of gelatine are colourless, but superficial ones are prominent and magenta in colour. On the surface of obliquely solidi- fied gelatine they form a beautiful magenta band with a metallic lustre. The gelatine is not lique- fied. Similar growths are obtained on agar and potato. It is one of the most striking of all the chromogenic bacteria. Cultures have the appearance of having been stained with an alco- holic solution of fuchsine. The colour varies in subcultures from magenta to a sealing-wax-red. Bacillus rubefaciens (Zirnmer- mann). — Rods '75 to 1/65 \L in length, ~3'2 p. in width, singly, in pairs, and in chains. Colonies are faintly reddish- yellow. The bacilli inoculated in the depth of gelatine grow along the track of the needle and form a greyish layer on the surface : later the jelly acquires a reddish tint. On agar the growth is grey and abundant. On potato the growth is at first grey, later reddish-brown, and the surface of the potato has a pink discoloration. They occur in water. Bacillus rubellus (Okada). — Rods resembling those of malignant oedema. They occur singly, in pairs, and filaments ; are motile, and possess flagella, and are often capsulated. They are anaerobic. Colonies are whitish, with off- shoots in the surrounding gelatine, which, later, is liquefied, and has a reddish tinge. The bacilli inoculated in the depth of gelatine produce a growth in the lower part of the needle track composed of isolated colonies with radiating processes. The jelly is liquefied, at first in the part corresponding with the growth, and later completely. The liquefied gelatine is coloured red. In agar the growth extends from below upwards, and the jelly is coloured red. In broth they grow rapidly. They were isolated from dust. Bacillus ruber (Breunig, Bacille rouye de Kiel. Laurent). — Rods 2*5 to 5 p. long, and •? to "8/n broad. Slightly motile. Colonies below the surface of gelatine are pale yellow, and super- ficial ones are blood -red. Inoculated in the depth of gela- tine they liquefy it and colour it bright red. There is also forma- tion of gas bubbles. 532 DESCRIPTION OF SPECIES. Potato is rapidly covered with a purplish-red growth. Broth be- comes turbid, and pink in colour. Milk is coagulated, and a blood- red colour develops on the surface and gradually extends. They were found in water. Bacillus rubescens (Jordan).— Rods 4 p in length, -9 p, in width, singly, in pairs, and short chains. Colonies pure-white. Inoculated in the depth of gela- tine there is a little growth in the track of the needle, and a pure- white prominent patch on the free surface. On agar the growth is white and shining, and later has a pink tinge. On potato the growth is flesh- coloured. Broth becomes turbid, and a scum forms on the surface. Milk after a time acquires a pink- ish colour. They occur in sewage. Bacillus rubidus (Eisenberg).— Rods and filaments. Colonies circular, granular, and slightly red. The bacilli inoculated in the depth of gelatine produce liquefaction and a brownish-red colour. On agar and potato they form a brownish-red growth, and liquefy blood serum. They occur in water. Bacillus sanguinis t y p h i (Brannan and Cheeseman).— Rods 1 to 2'5 p in length, -5 to '8 \L in width ; singly, in pairs, and in chains, and involution forms. Colonies granular, pale-brown. The bacilli inoculated in the depth of glycerine-agar produce a growth in the track of the needle composed of isolated, minute, white colonies. Ra'bbits inoculated die in from two weeks to a month. They were isolated from the blood of patients suffering from typhus fever. Bacillus saprogenes (Rosen- bach). — Three rod-formed organ- isms have been described by Rosenbach as intimately associ- ated with putrefactive processes. No. 1.— Large rods (Fig. 210), which form an irregular sinuous streak with a mucilaginous appear- ance when cultivated on nutrient agar-agar. Spore-formation pre- sent. They grow also very readily on blood serum, and all cultivations yield the odour of rotting kitchen refuse. They are not pathogenic. No. 2. — Rods shorter and thinner than No. 1. They develop very rapidly on agar-agar, forming trans- parent drops, which become grey. The cultivations yield a character- istic odour similar to the last. FIG. 210.— BACILLUS SAPROGENES, No. 1. (Rosenbach. ) They are pathogenic in rabbits. They appear to be identical with Bacillus fcetidus (Bacterium fceti- dum, Thin). They were isolated from a patient suffering from pro- fusely-sweating feet. No. 3. — See Bacterium sapro- genes. Bacillus scissus (Frankland).— Very short rods, 1 to 2 p in length, and 1 p. in width. Colonies yellowish, opaque in the centre, and periphery dentated. Inoculated in the depth of gela- tine there is no growth in the track of the needle, but a shining layer forms on the surface, and the jelly is coloured greenish. On agar the growth is shining and the jelly coloured green. On potato the growth is flesh- coloured. They occur in earth. Bacillus septicaemias haemor- rhagicae (p. 231). Bacillus septicus (Klein).— Rods varying in size. Non-motile. They form threads or leptothrix filaments, and are rounded at the ends. They are anaerobic, and form spores independently of access to air. DESCRIPTION OF SPECIES. 533 In a nourishing fluid they are overcome by the presence of micro- cocci, Bacterium termo or Bacillus subtilis. They occur in the soil, in putrid blood, and many putrid albuminous fluids, and occasionally in the blood- vessels of man and animals after death. Bacillus septicus acuminatus (Babes). — Rods with lancet-shaped ends, about the size of the bacilli of mouse-septicaemia. They exhibit polar staining. They can be culti- vated at 37° C. On agar and blood serum the colonies are circular, transparent, and later coalesce and form a yel- lowish layer. They are fatal to rabbits and guinea-pigs in from two to six days. They were isolated from an infant after death from septic infection occurring five days after birth. Bacillus septicus agrigenus (Nicolaier). — Rods resembling Ba- cillus septicaemias haemorrhagicse. Colonies circular, granular, with concentric zones of varying tints of brown. Intravenous injections are fatal to rabbits in twenty-four to thirty- six hours, and bacilli abound in the blood. They occur in recently manured soil. Bacillus septicus keratoma- laciae (Babes). — Short thick rods singly, and in pairs ; often capsu- lated. Colonies white, with dentated contours. The bacilli, inoculated in the depth of gelatine, grow in the track of the needle and on the surface ; gas bubbles are developed. On agar the growth is arbores- cent and opalescent. On blood serum they form a shining, somewhat transparent, dentated film. Cultures have an ammoniacal odour. They produce purulent inflam- mation of the cornea. They were isolated from the cornea in a case of septicaemia following keratomalacia in a child. Bacillus septicus ulceris gan- graenosi (Babes). — Short rods -5 to •6 p. in width. Inoculated in the depth of gela- tine they produce liquefaction and gas in the track of the needle. On agar greyish-white shining patches are found. On potato they develop a trans- parent film. They are pyogenic in rabbits and mice. They were isolated from the internal organs and blood in a case of septicaemia following gangrene. Bacillus septicus vesicae (Clado).— Rods 1-6 to 2 /t in length, •5 p. in width. Colonies circular, transparent, yellowish. The bacilli inoculated in the depth of gelatine form a delicate filament composed of closely-packed colonies, and on the surface there is a filmy growth. On agar they form a greyish- white layer, and on potato the growth is dry and brown. They are poisonous to rabbits, guinea-pigs, and mice. They were isolated from urine from a case of cystitis. Bacillus sessilis (L. Klein).— Rods resembling those of the hay bacillus. They are said to be distinguished by fission commencing in a newly formed rod before it has been set free from the spore. They were isolated from the blood of a cow. Bacillus smaragdino-phosplio- rescens (Katz). — Rods with pointed ends. 2 p in length, 1 /LI in width. Colonies circular, faintly yellow, with concentric rings. Inoculated in the depth of gela- tine a white filament forms in the track of the needle and a greyish- white patch on the free surface ; and there is sometimes liquefaction. In broth they produce turbidity. On potato they produce a thin brownish-yellow film. They are photogenic, and the 534 DESCRIFIION OF SPECIES. phosphorescence is most marked in cultures containing an excess of salt. They were isolated from a phos- phorescent herring. Bacillus smaragdinus fcetidus (Reimann). — Slender rods slightly bent. Colonies on agar irregular, with a yellowish granular nucleus, and transparent marginal zone. The bacilli inoculated in the depth of gelatine produce a growth in the track of the needle, and liquefaction at its upper part, and a greenish coloration. In the depth of agar the medium is coloured green. On potato the growth is brown. Cultures emit a strong odour. Intravenous injection produces death in rabbits in forty-eight hours. They were isolated from nasal mucus in ozsena. Bacillus solidus (Liideritz).— Rods 1 to 10 /A in length, -5 p. in width. They are anaerobic. Cultivated in grape-sugar gelatine they produce gas bubbles and a penetrating foul odour. The colonies are spherical, and in agar under a low power are seen to be composed of fine filaments like cotton wool. In broth with exclusion of oxygen they produce a copious growth with abundant formation of foetid gas. They were isolated from earth. Bacillus spiniferus (Unna).— Rods sometimes curved, 2 p. in length, -8 to 1 p in width, singly, in pairs, and masses. Colonies have peculiar spines, and later a radiated marginal zone. In the depth of gelatine minute, yellowish, isolated colonies develop in the track of the needle, and a furrowed, yellowish-grey patch on the free surface. On agar the same yellowish wrinkled growth appears. On potato they form a shining, faintly-yellow layer. They were isolated from the skin Rods sometimes bent, 3 to 8 p. in length, -6 n in width, and long filaments. Spore-formation present. They are anaerobic. Colonies are composed of fine radiating filaments, and liquefy the in eczema. Bacillus spinosus (Luderitz). — In agar the growth is composed of colonies of matted filaments, and there is gas-formation. They liquefy blood serum. They occur in earth. Bacillus stolonatus (Adametz), — Rods motile. Colonies on gelatine, circular, granular, and whitish, or yellowish- brown. Colonies on agar send off peculiar wavy processes. Inoculated in the depth of gela- tine a granular filament develops in the track of the needle, and a white patch on the free surface ; later the gelatine is excavated in the upper part, and the culture lines the cavity. On potato the growth is whitish. They occur in water. Bacillus stoloniferus (Pohl).— Rods 1'2 p. in length, '8 /* in width. Motile. Inoculated in the depth of gela- tine they produce rapid liquefaction ! in the track of the needle. On the surface of agar they form ; a white growth. On potato they grow abundantly, I but scarcely at all in milk. They occur in the water of marshes. Bacillus striatus albus (Bes- ser). — Rods sometimes bent. Colonies on gelatine appear as minute dry points. On agar the colonies have a brown 1 nucleus and clear marginal zone. On the surface of agar the bacilli produce a greyish- white thin layer. On potato the growth is trans- parent and slightly gelatinous. They occur in nasal mucus. Bacillus striatus flavus (Bes- ser). — Short rods, straight or curved ; involution forms. Colonies granular, yellowish. On the surface of agar they pro- duce a white growth, which later becomes sulphur yellow. DESCRIFHON OF SPECIES. 535 On potato a similar colour is produced. They were isolated from nasal mucus. Bacillus subflavus (Zimmer- mann). — Rods 1-5 to 3 p. in length, '11 fj. in width, and in chains. Motile. Colonies prominent, yellowish- white. On the surface of gelatine they form a yellowish-grey layer, and on the surface of agar and potato the growth is yellow. They occur in water. Bacillus subtilis (Hay bacillus). — Cylindrical rods as much as 6 /x in length. Single forms grow to double their length, and then undergo division. They also form threads which may be composed of FIG. 211. — BACILLUS SUBTILIS SPORES (BAUMGARTEN). long rods, short rods, and cocci. They are motile, and provided with a flagellum at each end. If the nourishing medium is impoverished, the multiplication of the rods by division gradually ceases, and spore- formation commences. The rods become motionless, and a dark spot is visible, either in the middle or towards one end. This gradually develops into a shining spore with a dark outline. The rods swell slightly during this process ; their contour becomes undefined, and soon disappears entirely ; spores being set free in about twenty- four hours. The spores are 1'2 p. long, and *G /x broad. They develop into rods in the following way : — On one side of the spore a swelling appears, at the summit of which an opening in the spore-membrane results, and the germ escapes ; this lengthens into a rod, and remains for a time attached to the empty spore-membrane. The spores are widely distributed, and occur in the air, soil, dust, etc. On the excrement of herbivorous animals the bacilli form a white efflorescence, and a thick crumpled skin on liquid manure. They flourish equally in liquids and upon damp, solid, nourishing media. They are aerobic ; depriva- tion of oxygen causes the growth of the bacilli to cease, and the rods degenerate. In plate-cultivations the colonies are white, and, under a low power, granular and irregular in outline and faintly-greenish. Liquefaction sets in, producing depressions like saucers. The centre is opaque, and is surrounded by a network of filaments, which extend into the gelatine surrounding the colony. FIG. 212. — PURE-CULTURE OF BACILLI'S SUBTILIS IN NUTRIENT GELATINE (BAUMGARTEN). Inoculated in the depth of gela- tine, liquefaction occurs rapidly in the track of the needle, and a film 536 DESCRIPTION OF SPECIES. floats on the surface. The liquefied gelatine, at first turbid, becomes clear as the bacilli settle at the bottom of the tube. On agar a wrinkled film develops, and also on serum. FIG. 213. PURE-CULTURE OF BACILLUS SUBTILIS ON THE SURFACE OF Nu- TRIENT AGAR. On potato the growth is white, and there is copious spore-forma- tion. On ordinary nutrient liquids they develop at first a thin, and subse- quently a thick, dense, crumpled pellicle, with copious spore-forma- tion. The simplest way to obtain a culture of the bacillus is to make a decoction of hay. The hay is chopped into small pieces, and boiled with distilled water in a flask for a quarter of an hour. The infusion is then filtered into a beaker, covered with a glass plate, and set aside in a warm place. In two or three days, the liquid swarms with the bacilli, the spores of which exist in great numbers in ordinary hay. A more sure method for obtaining a pure cultivation is as follows : — («) Add only a small quantity of water to some finely chopped hay, and set aside for four hours at 36° C. (6) Pour off the extract, and dilute it to the sp. gr. 1'004. (c) Boil gently for one hour in a bulb plugged with cotton wool. (d) Set aside 500 ccm. of the extract at 36° C. In about twenty-four hours, as a rule, a pellicle has commenced to develop upon the surface of the liquid. If the reaction is definitely acid, carbonate of soda solution must be added to the decoction. METHODS OF STAINING HAY BACILLUS. To demonstrate the flagella of the bacilli, they may be stained with hsematoxylin solution (Koch), or by Loffler's method. The linking together of cocci, long rods and short rods in the threads, is shown by treating with alcoholic solu- tion or f uchsine, or with iodine solution (Zopf). To stain the spores the cover-glass preparations must be heated to a very high temperature (210UC.), in the hot- air steriliser for half an hour, or they may be exposed for a few seconds to the action of concentrated sulphuric acid (Biichner), or floated for twenty minutes on hot solution of the dye. Bacillus subtilis similans.— There are several bacilli closely resembling Bacillus subtilis. Two have been isolated from human faeces by Bienstock which do not liquefy nutrient gelatine. No. I. Rods and filaments ; spore-formation present. On agar they produce a delicate wrinkled veil. No. II. Rods morphologically identical with No. I. On agar they produce a smooth, shining layer. Bacillus superficialis (Jordan). — Rods 2 '2 p. in length, and '1 p, in width ; singly, and in pairs. Motile. Colonies have a yellowish-brown DESCRIPTION OF SPECIES. 537 nucleus and transparent marginal zone. Inoculated in the depth of gela- tine there is a slight growth in the track of the needle, and after a time liquefaction at the upper part. On agar the growth is smooth and shining. In broth they produce turbidity. They will not grow on potato. They occur in sewage. Bacillus tennis sputigenus (Pansini). — Short rods, singly, and in pairs ; capsulated. They produce a whitish growth on the surface of gelatine. They coagulate milk. They are pathogenic in rabbits. They were isolated from sputum. Bacillus termo (Mace).— Thick rods 1*4 \t. long, and '8 p. wide, usually in pairs, sometimes in chains. Actively motile. Colonies whitish, with a grey edge surrounded by liquefied gelatine. Inoculated in the depth of gela- tine they form a funnel-shaped area of liquefaction, and later the whole of the jelly is liquefied. Broth is rendered turbid and a thin brittle pellicle is formed. They are associated with decom- position. Bacillus tetani (p. 457). Bacillus thalassophilus (Rus- sel). — Slender rods varying in length ; and filaments. They are anaerobic. Spore-formation pre- sent. Inoculated in the depth of gela- tine the growth appears in the lower part of the track of the needle in the form of cloudy colo- nies, liquefying the jelly and pro- ducing gas-bubbles. Cultures emit a penetrating odour. They were isolated from sea-mud. Bacillus thermophilus (Mi- quel). — Rods vary ing in size accord- ing to the temperature at which they are cultivated. In broth they grow best between 65° and 70° C., forming a copious deposit. They occur in air, soil, and water. Bacillus tremelloid.es (Tils).— Rods -75 to 1 /i in length, -25 p in width ; and in masses. Colonies circular, yellowish- brown. The bacilli inoculated in the depth of gelatine produce a growth composed of isolated yellow colo- nies in the track of the needle, and a yellow mass on the surface. They liquefy the gelatine. On agar the growth is slimy and golden-yellow. On potato they form an abund- | ant yellow growth. They occur in water. Bacillus tuberculosis (p. 378). Bacillus tuberculosis galli- narum (p. 402). Bacillus tumescens (Zopf). — Cocci, long and short rods. They i form a jelly-like disc *5 to 1 cm. in diam. on slices of boiled carrot, with the appearance of a rather tough, crumpled skin of a whitish I colour. Examination of this pel- j licle shows that it is formed of rows of rods lying closely together. These rods can be observed to divide into short rods and cocci. j Spore-formation occurs in two stages of development — viz., in the | cocci and in the short rods. A j cultivation is obtained by exposing ' slices of boiled carrot, slightly ! moistened, to the air at the tem- perature of the room. Bacillus typhi abdominalis (p. 342). Bacillus ubiquitus (Jordan).— Rods 1*1 to 2 p. in length, '1 /A I in width ; and filaments. Colonies -granular and well I defined. The bacilli inoculated in the | depth of gelatine produce a growth ! resembling that of Friedlanders pneumococcus. On agar and potato the growth i is greyish-white. They coagulate milk and reduce nitrates. They occur in air and water. Probably a variety of Bacillus candicans. Bacillus ulna (Cohn).— Cocci, short rods, long rods, and threads. Diam. of the cocci 1*5 to 2'2 p.. Spore-formation in both short and long rods. No septic odour is pro- 538 DESCRIPTION OF SPECIES. duced by this bacillus in a nourish- ing liquid. Cloudy masses are found on the surface of the liquid, which later form a thick dry pellicle, consisting of bundles of threads matted together. The for- mation of ellipsoidal spores occurs in the usual way ; they measure 2 '5 to 2 '8 \i long, and more than 1 /A wide. The bacillus is found in rotting eggs, and can be culti- vated on boiled white of egg. Bacillus ulna (Vignaf).— Rods 2 \L in length ; singly, and in pairs, and in short chains. Colonies composed of concentric zones varying in granularity. Inoculated in the depth of gela- tine, liquefaction occurs rapidly in the track of the needle ; later, there is a deposit at the bottom of the liquefied area and ji pellicle on the surface. On agar they form a white ad^ herent layer, and the jelly is tinged with brown. In broth a pellicle forms on the surface. On potato they form a pellicle with characteristic linear markings. They liquefy serum. Cultures produce a putrefactive odour. They occur in human saliva. Bacillus vacuolpsis (Sternberg). — Rods 1%5 to 5 p. in length, 1 /A in width, containing vacuolated proto- plasm; filaments, and involution forms. At times slowly motile. Inoculated in the depth of gela- tine, liquefaction occurs slowly at the upper part of the track of the needle, forming a cup-shaped cavity ; the liquefied gelatine is viscid, and a cream-white layer forms on the surface. In agar the development in the track of the needle is scanty ; on the surface a cream-white layer is formed, and the bacilli are united in long jointed filaments. On potato a similar growth is produced. They were isolated from the in- testine in fatal cases of yellow fever. Bacillus varicosus conjunctivae (Gombert).— Rods 2 to 8 /* m length, 1 p. in width. Inoculated in the depth of gela- tine they produce a greyish-white filament in the track of the needle, and a greyish-white patch on the surface ; liquefaction follows with- out turbidity. On the surface of agar a white, dry. adherent film is formed. On potato the growth is, at first, white and dry, later, reddish-brown. They produce hypersemia when injected into the conjunctiva. They were isolated from the healthy human conjunctiva. Bacillus venenosus (Vaughan). — Motile rods. Colonies circular, whitish. Inoculated in the depth of gela- tine there is growth in the track of the needle and on the free surface. On agar they form a white film, and on potato a moist brownish layer. They are pathogenic in small animals. They occur in water. Bacillus venenosus brevis (Vaughan). — Rods short and thick. Colonies are yellow and composed of concentric rings. Inoculated in the depth of gela- tine they grow in the track of the needle and over the free surface. On agar they produce a white film. On potato the growth is brownish. They are pathogenic in small animals. They occur in water. Bacillus venenosus invisibilis. — Slender rods. Colonies irregular, granular. Inoculated in the depth of gela- tine the growth is extremely slow both in the track of the needle and on the surface. On agar there is a whitish film, and on potatoes a brownish layer. They are pathogenic in small animals. They occur in water. Bacillus venenosus lique- faciens (Vaughan).— Rods. CoJonies circular, granular, yel- lowish. Inoculated in the depth of gela- tine they grow in the track of DESCRIPTION OF SPECIES. 539 the needle and on the surface, and liquefaction occurs after some weeks. On agar they produce a white growth, and on potato it is brown- ish or yellowish. They are pathogenic in small animals. They occur in water. Bacillus ventriculi (Raczyn- sky). — Rods 1-5 to 3 p in length, 1 /z in width, singly, in pairs, and in short chains. Colonies have a dark nucleus and transparent periphery. On agar they form a white layer. They were isolated from the digestive tract of dogs. Bacillus vermicularis (Frank- land). — Large bacilli 2 to 3 /* in length, 1 /z in width, and long threads. Spore-formation present. Colonies are irregular in contour, the irregularity increasing as the colony comes to the surface. The peripheral part is composed of closely packed, wavy bands of bacilli, and the centre is irregular and wrinkled. The bacilli inoculated in. the depth of gelatine form a flattened band in the track of the needle, and a grey layer on the surface ; lique- faction slowly follows. On agar they produce a smooth,, shining, grey layer, and on potato a thick, irregular, flesh-coloured growth. They reduce nitrates. They occur in water. Probably identical with Bacillus vermicu- losus. Bacillus vermiculosus (Zimmer- mann). — Rods 1'5 /z in length, -85 p in width, singly, in pairs, very short chains and long filaments. They are slowly motile. Colonies irregular ; grey, granu- lar. Inoculated in the depth of gela- tine they produce, after four days, liquefaction in the upper part of the needle track, which spreads downwards, and a reddish-grey sedi- ment collects at the bottom of the liquefied area. On agar the growth is smooth and shining, and on potato yellowish- grey. They occur in water. Bacillus violaceus (vide Bacil- lus ianthinus). Bacillus violaceus Laurentius (Jordan).— Rods 3 to 3'6 p, in length, '7 p in width. Colonies violet, surrounded by liquefied gelatine. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle, and a violet sediment collects at the bottom. On agar the growth is violet, later black. On potato there is a copious growth, changing in colour from violet to black. In broth a violet colour is pro- duced in the presence of nitrates. They coagulate milk, and render it bluish-violet. They occur in water. Probably identical with Bacillus ianthinus (Zopf). Bacillus virescens (Frick).— Rods and filaments. Colonies irregular, granular, green. On the surface of gelatine they colour the medium green. They grow on agar. On potato they form a brownish growth. In broth a pellicle is formed on the surface, and beneath it the liquid is coloured green. They were isolated from green sputum. Bacillus viscosus (Frankland). — Rods 1*5 to "2 p. in length, singly and in pairs. Colonies granular, with hairlike processes extending into the gela- tine, which is liquefied and has a green colour. Inoculated in the depth of gela- tine they produce liquefaction and a green fluorescence. On agar they form a greenish- j white layer, and colour the jelly I green. On potato the growth is brown. Probably identical with Bacillus fluorescens liquefaciens. Bacillus Zurnianus (List).— \ 8 R A OF THW UNIVERSITY o40 DESCRIPTION OF SPECIES. Rods 1-2 to 1 -5 p in length, -6 to -8 A* in width. Colonies greyish-white, viscid. The bacilli inoculated in the depth of gelatine develop slightly in the track of the needle, and pro- duce a prominent grape-like growth on the free surface. On potato the growth is grey or tinged with yellow. They occur in water. Bacterium aerogenes (Miller). —Short rods, singly and in pairs. Motile. The colonies are circular, well defined, and yellowish. Inoculated in the depth of gela- tine the growth in the track of the needle is brownish-yellow, and a flat greyish button forms on the free surface. On agar a pulpy layer develops. On potato the growth is pulpy and yellowish-white. The bacteria possess great power of resisting the effect of acids. They were isolated from the diges- tive tract. Bacterium brunneum (Schroter). — Motile rods, produ- cing a brown colour. They were observed on a rotting infusion of maize. Bacterium decalvans (Thin).— Cocci, singly or in pairs, 1-6 //. in length. They were observed in the roots of the hair in cases of Alopecia areatti. Bacterium fusiforme (Warm- ing).—Rods spindle-shaped, with pointed ends, 2-5 p. long, and '5 to •8 n thick. They were described as forming a spongy layer on the surface of sea-water. Bacterium gingivae pyogenes (Miller).— Short rods. The colonies are circular and rapidly liquefy gelatine. The bacteria inoculated in the depth of gelatine produce rapid liquefaction in the track of the needle and a white sediment. On agar they produce a moist white growth. They are pyogenic when inocu- lated subcutaueously in small ani- mals, and cause a fatal result when injected to the peritoneal cavity. They occur in the deposit on the teeth. Bacterium hyacinthi (Wakker). — Cocci resembling Bacterium termo. They were observed in the yellow slime of diseased hyacinth bulbs. Bacterium hydrosulfureum ponticum (Zelinsky-). — Long motile rods. On agar a dark coffee-coloured pigment is produced, which turns black when exposed to air. Cultures give off sulphuretted hydrogen. They were isolated fromdredgings in the Black Sea. Bacterium litoreum (Warming). — Cocci ellipsoidal, 2 to 6 /* long, 1'2 to 2-4 fj, wide ; singly, never as chains or zoogloea. They occur in sea-water. Bacterium luteum (List).— Rods from I'l to 1'3 ^ long. Non- motile. The colonies are slimy, with orange centres. Inoculated in the depth of gelatine an orange growth occurs, principally at the point of puncture. Milk is coagulated. They occur in water. Bacterium merismopedioides (Zppf).— Threads I to 1-5 w in thickness ; these subdivide into long rods, short rods, and finally into cocci. The cocci divide first in one and subsequently in two directions, forming characteristic groups, which appear like merismo- pedia. These groups may eventually consist of 64 by 64 cells or more, and ultimately form zoogloea. The cocci develop again into rods and threads. They were observed in water containing putrefying substances (River Panke, Berlin). Bacterium navicula (Reinke and Berthold). — Cocci spindle-form or ellipsoidal, including motile and non-motile forms. They have one or more dark spots, which may be coloured blue by iodine. They have been observed in rot- ting potatoes. DESCRIPTION OF SPECIES. 541 Bacterium photometricum(En- gelmann). — Rods slightly reddish in colour ; motile. The movements are stated to depend on light. Bacterium synxanthum (Ehren- berg ; Bacterium xanthinum ; Bac- terium of yellow milk). — Cocci '7 to 1 /z in length, and rod-forms. They produce a yellow colour in boiled milk, which at first becomes acid, and then strongly alkaline. They also occur on boiled potatoes, carrots, etc., where they form small lemon-yellow masses. The colouring-matter is soluble in water, insoluble in ether and alcohol, unchanged by alkalies, de- colorised by acids. It is similar to yellow aniline colours, both spectroscopically and in ordinary reactions. Bacterium termo (Vignal).— Rods 1*5 to 2 p. in length, *5 to '7 p. in width. Colonies white, surrounded by liquefied gelatine. The bacilli inoculated in the depth of gelatine produce a funnel- shaped area of liquefaction ; later, the jelly is completely liquefied and coloured green. Cultures have a strong putrefactive odour. In broth they form a white deposit and colour the medium green. They were isolated from human saliva. Bacterium tholoeideum (Gess- ner). — Rods similar to Bacillus lactis aerogenes. Pathogenic in small animals. They were isolated from healthy human evacuations. Bacterium ureas (Cohn).— Cocci 1*25 to 2 /z in diam., singly or in chains, and rods. The rods split up by division into chains of cocci, which after a time are set free. The cocci increase further by subdivi- sion, and a jelly-like membrane develops around them. Masses of cocci exist in the form of irregular or roundish lumps. They are aerobic. Cultivations, after twenty-four hours, consist exclusively of rods ; after forty-eight hours, of cocci chains ; and in fourteen days, of zoogloea ; the cocci transplanted into fresh nourishing solution again grow into rods. These observations point to the existence of a pleo- morphic species, Bacterium urece ; and the former nomenclature, Micro- coccus urece, must be regarded as untenable. In urine they set up ammoniacal fermentation, converting urea into carbonate of ammonia. Rods, 2 p. long and 1 /z wide, have been iso- lated from stale urine (Bacillus urese, Leube), which also most energetically cause the ammoniacal fermentation of urine. Bacterium ureae (Jaksch).— Rods 2 /* in length, 1 /z in width. Colonies on gelatine semi-trans- parent. Inoculated in the depth of gela- tine the bacilli form a delicate branching growth in the track of the needle. They convert urea into carbonate of ammonia, and cultures smell of herring brine. They occur in ammoniacal urine. Bacterium yiolaceum (Bergon- zini). — Rods similar to Bacterium termo, '6 to 1 /z thick, 2 to 3 tt long. They occur on white of egg, forming a violet pigment. Bacterium Zopfii (Kurth).— Cocci, 1 to 1'25 /z in diam. ; rods and threads. Cultivated in a streak on nutrient gelatine spread out on a glass slide, a peculiar develop- ment takes place. In twenty-four hours after inoculation threads have developed ; in forty-eight hours windings of the threads are observed, and in six days the threads have broken up into cocci. They were observed in the intestine of fowls, especially in the contents of the vermiform appendix. In- oculation of rabbits was followed by negative results. Identical with Bacillus figurans (Crookshank). Beggiatoa alba (Vauch).— Cocci,, rods, spirals and threads (Fig. 215). The threads are indistinctly articu- lated, actively oscillating, and colour- less ; their protoplasm contains o42 .DESCRIPTION OF SPECIES. numerous strongly refractive gran- ules consisting of sulphur. They occur as greyish or chalk- white gelatinous threads, 3 to 3 '5 /* thick, in sulphur springs and marshes. Beggiatqa mirabilis (Cohn).— Threads distinguished by their breadth, which may reach 30 p. They are motile, bent and curled in various ways, and rounded at the ends. Around the threads, isolated cells have been observed, families, bound together by gela- tinous substance. Later they be- come larger, globular or ovoid in shape, and hollow, containing watery fluid in their interior. The families reach a diameter of 660 ju, in which the cocci form simply a peripheral layer. The hollow fami- lies or vesicles are often perforated, presenting a delicate reticulated ap- pearance, which finally may become broken up into irregular structures. The red colouring-matter can be t>Hf Z°P1?\ S1™CE88IY* CHANGES IN THE SAME THREAD, thread form ; b, breaking up into rod forms ; c, into cocci (Kurth). macrococci, but spiral forms are as yet unknown. The threads are filled with sulphur granules. They occur in sea-water, forming a white gelatinous scum on decomposing algae. Beggiatoa r oseo-p e r s i c i n a {Co/mm roseo-perttdna. Bacterium rubescens, or Peach-coloured bac- tfirtum, Lankester).— Cocci, rods, spirals, and threads (Fig. 216). The «occi, globular or oval, reach 2-5 /* an diam. They form at first solid distinguished from other red pig- ments, and it is designated by the name bacterio-purpurin. It is quite distinct from the pigment produced by Micrococcus prodigiosus, being peach-blossom red, and insoluble m water, alcohol, etc. Examined spectroscopically, it shows a strong absorption in the yellow, and a weaker band in the green and blue, as well as a darkening in the more refrangible half of the spectrum. In the cocci, especially of the older DESCRIPTION OF SPECIES. 543 vesicles, dark granules are to be seen, which consist of sulphur. The micro-organisms occur on the surface of marshes, or on water in which-- 1 algge are rotting. They form a rose-red, blood-red, violet- red, or violet-brown scum ; and sometimes in such quantity that Cladothrix dichotoma (Cohn). — Threads resembling those of lep- tothrix ; slender, colourless, not articulated, straight or slightly undulated, and in places twisted in irregular spirals with pseudo- branchings. The development can be traced from the cocci to rods and FIG. 215.— BEGGIATOA ALBA. A. Threads at base distinctly linked, partly spiral. B. A -thread, spiral in its whole length. C, D. Fragments detached from threads ; immotile. E. Active spirillum-forms, with a flagellum at either end. F, G. Thin and short spiral forms. H. A spiral showing the individual links, x 540 (Zopf). whole marshes and ponds may be coloured blood-red by them. Spirillum sanguineum, rosaceum, violaceum, monas vinosa and Okenii, and Rhabdomonas rosea are pos- sibly phase-forms of Beggiatoa roseo-persicina. threads. The latter are at the beginning simple threads, which were formerly described as Lepto- thrix parasitica, or, if coloured by impregnation with iron, as Lepto- thrix ochracea. Later they form false branches by single rods turning 544 DESCRIPTION OF SPECIES. aside, which by repeated division lengthen into threads. A thread appears to be first composed of long rods, then of short rods, and lastly of cocci. The iodine reaction must be applied to distinguish these forms, especially when the sheath of the threads has a yellow, rust- red, olive-green, or dark brown coloration. The cocci may grow into rods while still in the sheath, and finally become leptothrix threads, surrounded by a delicate gelatinous sheath, from which the media small tufts, about 1 to 3 /LI, and floating masses. Cladothrix Fbrsteri (ride Strep- tothrix Forsteri, Cohn). Cladothrix intricata (Kussell). — Rods and filaments. Colonies are composed of a net- work of twisted threads. Inoculated in the depth of gela- tine fine filaments spread out from the track of the needle, and the gelatine is liquefied. Grown on agar the filaments penetrate the jelly. FIG. 216.— PHASE-FORMS OF BEGGIATOA ROSEO-PERSICINA (WARMING). false branching proceeds. Frag- ments may break oft3, which are actively motile, and appear as vibrios, spirilla, and spirochaeta- forms. They may also occur in zooglcea (Fig. 217). They are the commonest of all bacteria in both still and running water, in which organic substances are present. They are observed also in the waste water of certain manufactures, such as sugar. Arti- ficially they can be cultivated on infusions of rotting algae and ani- mal substances, forming on these In broth the growth is abundant. They were isolated from sea dredgings. Cladothrix invulnerabilis (Ac- osta, y Grande Rossi). — Filaments which produce in gelatine a white thread, and liquefy it very slowly. On potato the growth is abundant and chalky in appearance. In milk they form a firm yel- lowish pellicle ; and in broth and in water the growth is abundant. They occur in water. Clostridium butyricum (vide Bacillus bitty ricus). DESCRIPTION OF SPECIES. 545 Clostridium foetidum (Libo- rius). — Rods 1 /j, in width, singly and in filaments. Spore-formation re- sembles that of Bacillus butyricus. They are anaerobic. Colonies rapidly liquefy gelatine. gas-formation with unpleasant smell and splitting up of the jelly. They were isolated from earth. Crenothrix Kiihniana (Raben- horst). — Cocci, rods, and thread- forms. The cocci are globular, FIG. 217.— CLADOTHRIX DICHOTOMA. A. Branching schizomycete :— (a) Vibrio-form ; (6) Spirillum-form [slightly mag- nified]. B. A screw-form with («) Spirillum -form ; (b) Vibrio-form. C. Long spirochaeta-form. I). Fragment with spirillum-form at one end, vibrio-form at the other. E. Screw-forms : — (a) continuous; (b) composed of rods; (c) composed of cocci. F. Spirochseta-form : — (a) continuous; (b) composed of long rods; (c) short rods; (d) cocci (Zopf). On agar the colonies form branch- ing processes resembling colonies of Bacillus cedematis maligni. Inoculated in the depth of gela- tine liquefaction spreads from be- low upwards. There is abundant 1 to 6 n in diam. The threads are colourless, 1*5 to 5 p. thick, and club- shaped at the extremity, reaching a diam. of 6 to 9 p. The threads form colonies with a brick -red, olive- green,or dark-brown to brown- black 35 546 DESCRIPTION OF SPECIES. coloration, caused by impreg- nation with oxide of iron. The threads are distinctly articulated, and ensheathed. The segments are set free when the sheath bursts, and develop into new threads. In other cases the segments remain enclosed, and subdivide into discs, FIG. 218.— CRENOTHRIX KUHNIANA. «, 6, c, d, e. Cocci in various stages of fission, x 600. /. Zooglcea of cocci, x 600. (j. Various forms of zooglcea, natural size. h. Colony of threads composed of rods grown out of a zooglcea of cocci. i — r. Thread-forms ; some straight, others spiral, with more or less differentia- tion between base and apex, (r) is composed of short rods at the base, and above these of cylindrical segments, and at the apex these segments have divided into cocci, x 600 (Zopf ). DESCRIPTION OF SPECIES. 547 which, by vertical fission, break up into globular forms (cocci). These again develop into new threads, either within the sheath, eventually penetrating it, or after they are set free. The micro-organism appears in little whitish or brownish tufts in wells and drain-pipes, and it not only renders drinking-water foul, but may stop up the narrower pipes. Diplococcus albicans ampins (Bumm). — Cocci resembling gono- cocci but much larger, singly and in tetrads. Colonies are prominent and greyish-white. In the depth of gelatine they produce a greyish-white growth in the track of the needle and on the free surface. They slowly liquefy the gelatine. They were obtained from the vaginal mucous membrane. Diplococcus albicans tardis- simus (Bumm). — Cocci morpho- logically identical with gonococci. They grow extraordinarily slowly on gelatine. They form very minute colonies, which are opaque and greenish- brown in colour. Inoculated in the depth of gela- tine isolated greyish- white colonies develop in the track of the needle, and on the free surface a thin, white, waxy film with dentated edge. On agar the growth is very similar. They were isolated from the vaginal mucous membrane. Diplococcus citreus conglome- ratus (Bumm). — Cocci in pairs resembling gonococci, 1 '5 \L in diam., in tetrads and in masses. Colonies lemon-yellow ; irregular in form. Inoculated in the depth of gela- tine the cocci develop in the track of the needle, and liquefaction com- mences at its upper part. The growth on the free surface is yellow, and floats on the liquefied gelatine or subsides to the bottom of the liquefied area. They are present in gonorrhoeal pus, in air and in dust. Diplococcus citreus lique- faciens (Unna).— Oval cocci -4 to •1 /i in diam., in pairs, tetrads, short chains, and masses. Colonies appear in the form of circular discs, at first greyish- white, later lemon-yellow. They are finely granular, and have sharply defined contours. Inoculated in the depth of gela- tine, at the end of a week the growth is found on the free surface, forming a shining yellow layer ; in two weeks liquefaction commences, and the growth floats on the liquefied gelatine which is also yellowish and turbid. On the surface of agar a yellow- ish-brown layer is rapidly formed. The appearance is similar on potato. They were isolated in cases of eczema seborrhosicum. Diplococcus coryzae (Hajek).— Large diplococci. Colonies are white, prominent. Inoculated in the depth of gela- tine the growth resembles the pneumococcus. On agar a white layer is formed. They are probably identical with Friedliinder's pneumococci. They were isolated from the mucus in acute nasal catarrh. Diplococcus flavus liquefaciens tardus. — Cocci resembling gono- cocci. Colonies are circular, shining, and chrome-yellow in colour. Inoculated in the depth of gela- tine a yellowish growth occurs along the needle track, and also on the free surface. In a month the surface is depressed, but the gelatine is not liquefied until about two months have elapsed. On agar a yellowish-white layer is formed, and on potato the colour is more pronounced. They were isolated from the skin in eczema seborrhoeicum. Diplococcus fluorescens foeti- dus (Klamann). — Cocci in pairs and chains. Colonies circular, forming a 548 DESCRIPTION OF SPECIES. brownish deposit surrounded by liquefied gelatine which has a violet or greenish tinge. Inoculated in the depth of gela- tine they produce liquefaction along the track of the needle, with a hemispherical excavation of the gelatine at the upper part. An iridescent film floats on the surface and a greenish sediment forms at the bottom of the liquefied area. On agar the layer is brownish. On potato the growth is granular, and the potato in the vicinity has a bluish colour. They were cultivated from the nasal mucus. Diplocqccus inter cellularis meningitidis (Weichselbaum).— Cocci singly, in pairs, tetrads and masses. They grow at 37°C. Colonies on agar are granular and yellowish-brown. On the surface of agar they form a greyish-white viscid growth. In the depth of agar the growth only occurs in the upper part of the needle track. On blood serum and broth there is very little growth, and none on potato. Cultures quickly lose their vitality. They are pathogenic in mice, guinea-pigs, rabbits, and dogs. They were isolated from the exu- dation in cases of cerebro-spinal meningitis, and were observed in the interior of pus cells. Diplpcoccus luteus (Adametz). — Cocci 1'2 to 1*3 p in diam., singly and in chains. Motile. Colonies are circular and slightly yellow, and granular. Old colonies are bright yellow. On the surface of gelatine a growth occurs in concentric circles of a lemon-yellow colour, and the gelatine is coloured reddish-brown. After several weeks liquefaction sets in. On agar a yellow layer forms, and the jelly is coloured reddish-brown. On potato the growth changes from yellow to brown. Milk is coagulated. They were obtained from water. Diplococcus of pneumonia in horses (Schutz). — Oval cocci, singly or in pairs, capsulated. Colonies small and white. In the depth of gelatine a row of colonies develops in the track of the needle. On agar the growth is composed of transparent droplets. Injection into the lung is said to produce pneumonia, ending fatally in eight or nine days. They are pathogenic in rabbits, guinea-pigs, and mice. They were isolated from the lungs of a horse suffering from pneumonia. Diplococcus roseus (Bumm). — Cocci identical in description with gonococci. Colonies are pink, granular, and irregular in form. Inoculated in the depth of gela- tine the cocci grow freely in the track of the needle and on the surface, developing a pink colour and slowly producing liquefaction. They are present in the air. Diplococcus subflavus (Bumm). — Diplococci similar to gonococci. Colonies greyish- white, later yellow. They grow in gelatine and on blood serum, and liquefy broth. They produce suppuration when injected subcutaneously in man. They were isolated from lochial discharges, the vesicles of pem- phigus, and from the secretion in colpitis in children. They stain by Gram's method. Hasmatococcus bovis (Babes).— Cocci oval, singly, in pairs, and in masses. Inoculated in the depth of gela- tine minute colonies develop in the track of the needle. On agar the growth is composed of transparent droplets. On potato a yellowish shining film is formed. On blood serum the growth is similar to that on agar. They produce a fatal result in rabbits and guinea-pigs in a week or ten days. They were isolated from the DESCRIPTION OF SPECIES. 549 blood and organs of cattle which died of an epidemic disease asso- ciated with haemoglobinuria. Helicobacterium aerogenes (Miller). — Bacilli singly, in chains and long wavy filaments. Motile. Colonies whitish, varying in form. Inoculated in the depth of gela- tine the bacilli give rise to a faintly yellow growth in the track of the needle, and an almost invisible, rapidly growing layer on the surface. On potato the growth is dry and brownish. They were isolated from the healthy intestinal tract. Leptothrix buccalis (Robin). — Long, thin threads, -7 to 1 /x broad, colourless, often united in thick bundles or felted together. Masses of cocci occur with the threads, and the threads themselves are com- posed of long rods, short rods, and cocci. The threads may break up into spiral, vibrio, and spirochaeta forms. The last-named occur in large numbers in the mouth, and have been named Spirochceta buc- calis. Leptothrix buccalis is found in teeth slime, and is believed to be intimately connected with dental caries. The threads pene- trate the tissue of the teeth, after the enamel has been acted upon by acids generated by the fermentation of food. The short rods, long rods, cocci, leptothrix-forms, and screw- forms are found in the dental canals. The threads of Leptothrix buc- calis have a special staining reaction (Leber). They become coloured if placed in an acid medium with iodine ; if the medium be alkaline, it must first be acidified with very dilute hydrochloric acid or acetic acid. The contents are stained violet, and contrast with the sheath and septa, which remain uncoloured. Leptothrix buccalis (Vignal).-— Rods 1 to 1-5 \i in width, 1-6 to 30 /u. in length. Colonies greyish-white, promi- nent and furrowed. Inoculated in the depth of gela- tine a filament forms in the track of the needle, and a growth occurs on the free surface. Liquefaction sets in at the upper part, forming a cup-shaped cavity, and a bluish skin floats upon the liquid. The liquefaction gradually extends to the side of the tube, and a deposit is formed at the bottom of the liquefied gelatine. On agar the layer is white, wrinkled and transparent, and later yellowish. In broth there is turbidity, but no skin on the surface. On potato the growth is greyish- white. They are occasionally present in the mouth in health, and are possibly identical with lepto- thrix buccalis (Robin). Leptothrix gigantea (Miller). — Long rods, short rods and cocci can be observed in the same thread. There are also screw-threads, which may take the form of spirals, vibrios, or spirochaetae. The threads increase in diameter from base to apex ; corresponding with the thickness of the threads, the rods and cocci show different dimensions. They have been observed in the diseased teeth of dogs, sheep, cats and other animals. Leuconostoc mesenteroides, Cienkowski (Gomme de siicrerie, Froschlaichpilz, Frogspawn fungus). — Cocci and rods singly, in chains, and in zoogloea, surrounded by a thick gelatinous envelope. The life- history has been very thoroughly investigated. The spores, 1*8 to 2 p. in diam., are of a round or ellipsoidal form, with thick mem- brane and shining contents. The outer membrane-layer bursts, and a middle lamella oozes out, and forms a thick gelatinous envelope, while the inner layer remains adherent to the plasma. Thus the spore- germination leads to the formation of a coccus with a gelatinous en- velope. The coccus then elongates into a short rod-form, and the gelatinous envelope becomes ellip- soidal. The rod divides into two cocci, and each of these lengthens into a rod and divides. By repetition 550 DESCRIPTION OF SPECIES. of this process a chain of cocci results, encased in a cylindrical or ellipsoidal envelope. The chains increase in length, become twisted up, and eventually fall apart into pieces of various lengths. In nourishing liquids a great number of little masses are formed, which adhere together, and produce pseudo-parenchymatous structures. These latter may join together, forming still larger agglomerations. This micro-organism occurs occa- sionally in beet-root juice and the molasses of sugar-makers, forming large gelatinous masses resembling frog-spawn. The vegetation is so rapid that forty-nine hectolitres of molasses, containing 10 per cent, of sugar, were converted within twelve hours into a gelatinous mass ; consequently, it is a for- midable enemy of the sugar manu- facturers. FIG. 219.— LEUCONOSTOC MESENTEROIDES. 1. Spores. 2. Spores after germination, showing gelatinous envelope. 3, 4, 5, 6. Increase by division. 7. Glomerular form of zooglcea. 8. Section of an old kowskif Z°°gloea" 9* Cocci chains with arthrospores (Tieghem and Cien- The masses of zooglcea are of almost a cartilaginous consistency, and admit of sections being made with a razor. After a long time the envelope liquefies, and the cocci are set free ; the latter introduced into fresh nourishing media develop new colonies. In the chains some of the cocci become enlarged with- out changing their form. These acquire the properties of spores, and are firthrospores. Micrococcus acidi lactici (Marpmann). — Large cocci, singly and in pairs. Colonies yellowish-white. On the surface of gelatine the cocci produce a yellow layer. They grow in milk, producing a reddish colour, and coagulation due to the formation of lactic acid. They were isolated from milk. Micrococcus acidi lactici lique- faciens (Kreuger).— Cocci oval, 1 DESCRIPTION OF SPECIES. 551 to 1*5 p. in diam., in pairs and in tetrads. Colonies white. Inoculated in the depth of gela- tine the cocci produce a granular filament, changing in two or three days to liquefaction in the form of a funnel ; later, a wrinkled mem- brane floats on the surface of the liquid. In milk they produce lactic acid. They were isolated from butter which had turned cheesy. Micrococcus aerogenes (Miller). — Oval cocci. Colonies are dark and regular in contour, but have a peculiar spotted appearance. Inoculated in the depth of gela- tine a brownish-yellow growth occurs along the track of the needle, and on the free surface a white button-like elevation. After a time the gelatine is slowly liquefied. On agar a yellowish- white pulpy layer forms, and a similar growth appears on potato. They resist the action of acids, so that the presence of gastric juice does not impede their develop- ment. They were obtained from the intestine. Micrococcus agilis (Ali-Cohen). —Cocci 1 /x in diam., singly, in pairs, tetrads, and in chains. They are motile, and possess flagella. Inoculated in the depth of gela- tine they grow in the track of the needle, and produce, after two or three weeks, liquefaction or excava- tion of the jelly. On agar and potato the growth is pink. They occur in water. Micrococcus agilis citreus (Menge). — Cocci in pairs, chains and masses. They are motile, and each coccus possesses a single fla- gellum. Colonies appear surrounded by clouded gelatine. Inoculated in the depth of gela- tine there is a scanty growth in the track of the needle, and on the sur- face a bright yellow patch. On agar they form a yellow layer, which is viscid, and may be drawn out in long threads. In broth they produce cloudiness and a viscous deposit. The growth on potato is bright yellow. Milk is not coagulated. They were isolated from an in- fusion of peas. Micrococcus albus liquefaciens (Besser). — Large cocci in chains and in masses. They are anaerobic. Colonies on agar exhibit concen- tric rings of different shades of brown. Inoculated in the depth of gela- tine they produce liquefaction in the track of the needle. They occur in mucus from the nose. Micrococcus amylivorus (Bur- rill). — Oval cocci, 1 to 1'4 p. long, •7 n broad, singly, in pairs, and rarely in fours, never in chains, are found embedded in an abundant mucilage which is very soluble in water. They have been described as pro- ducing the so-called " fire blight " of the pear tree and other plants. Micrococcus aquatilis (Bolton). — Small cocci in masses. Colonies circular, prominent, and pure-white. Inoculated in the depth of gela- tine, there is a white growth in the track of the needle and also on the free surface. On agar the growth is white. They occur in water. Micrococcus aquatilis invisi- bilis (Vaughan). — Cocci oval. Colonies brown. In gelatine there is a slight growth in the track of the needle, and a more abundant growth on the free surface. On agar they form a white film. On potato the growth is in- visible. They occur in water. Micrococcus aurantiacus (Cohn). — Cocci spherical or oval, 1-3 to 1-5 p. in diam., singly, in pairs, and in groups. Colonies orange-yellow. Inoculated in gelatine they form minute colonies in the track of the 552 DESCRIPTION OF SPECIES. needle, and a prominent hemi- spherical yellow growth on the free surface. On agar the growth is orange- yellow, and on potato yellow and slimy. They occur in water. Micrococcus botryogenus (Johne, Rabe). — Cocci 1 to 1'5 p, in diam., in wavy chains. Colonies circular, sharply defined. At first silver-grey, later yellowish- grey with metallic lustre, they produce an odour like that of strawberries. Inoculated in the depth of gela- tine a greyish- white filament de- velops, with slight liquefaction of the gelatine ; later, it becomes milk-white, and at its upper part a characteristic bubble appears. They make hardly any growth on agar. On potato they grow very abun- .dantly, forming a yellowish layer with the same odour as the colonies on plate cultivations. Inoculated guinea-pigs die of septicaemia ; in sheep and goats severe inflammation spreads from the point of inoculation. Mice are immune. In horses an inflamma- tory oedema is at first set up, followed in four to six weeks by the formation of new growths, which sometimes suppurate and contain large numbers of micro- cocci. They were found in tumours of the spermatic cord and of the connective tissue in other parts in horses. Micrococcus candicans (Fliigge). — Cocci which collect in masses. In plate-cultivations they form in two or three days milk-white colonies ; while those below the surface of the gelatine are yellow- ish. Under a low power the deep colonies are quite circular, with smooth margins, of a blackish-brown colour, and very slightly granular ; the superficial colonies are quite irregular in outline, and are finely granular. Cultivated in test-tubes they form a white nail-shaped cultivation. They were isolated from contami- nated plate-cultivations. They occur in the air. Micrococcus candidus(Cohn).— Cocci forming snow-white points and spots upon slices of cooked potato. Micrococcus carneus (Zimmer- mann). — Cocci *8 /z in diam., occur- ring in masses. Colonies circular, greyish-white, with the centre tinged with red. Inoculated in the depth of gela- tine they form a white, granular filament in the track of the needle, and a pale pink layer on the free surface. On the surface of oblique gela- tine a flesh-coloured layer develops, which later assumes a violet colour. On agar the growth is similar. On potato the growth is abundant and red in colour. They were isolated from water. Micrococcus cerasinus siccus (List). — Cocci *25 to '12 p in diam., singly and in pairs. They can best be cultivated at 37° C. On agar they form a cherry-red layer, and a similar growth on potato. The pigment is insoluble in alco- hol, ether, and water, and is not destroyed by acids or alkalies. They occur in water. Micrococcus cereus albus (p. 178). Micrococcus cereus flavus (p. 178). Micrococcus cinnabareus (Fliigge). — Large cocci occurring in twos, threes, and fours. Colonies develop very slowly, and are punctiform, and bright red at first, and afterwards reddish-brown. The cocci inoculated on the sur- face of gelatine form a heaped-up, red-coloured growth. They were found contaminating old cultivations. Micrococcus citreus (List).— Cocci 1'5 to 2-2 /j. in diam., singly, in pairs and chains. Colonies are irregular in form, moist and shining, and yellowish in colour. DESCRIPTION OF SPECIES. 553 In the depth of gelatine the growth is very scanty. On the surface of agar the growth is yellowish. On potato the growth is similar but more abundant. Micrococcus concentricus (Zimmermann). — Cocci -9/zin diam., in masses. Colonies bluish-grey. Inoculated in the depth of gela- tine there is no growth in the track of the needle, but concentric rings form on the free surface. On agar the growth is greyish- white and smooth. On potato yellowish and slimy. They occur in water. Micrococcus cremoides (Zim- mermann).— -Cocci '8 /z in diam., occurring in masses. Colonies are spherical, granular and yellowish. The margins are dentated or irregular, and processes extend into the surround ing gelatine. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle in a few days. A yellow growth floats on the liquefied gelatine, and a yellowish mass subsides to the bottom of the liquid. On agar a smooth shining layer is formed, and on potato the growth is abundant. They occur in water. Micrococcus crepusculum (Cohn. Monas crepusculum, Ehren- berg. Jfikrokokken in faulenden Substraten, Fliigge). — Round or short oval cells, scarcely 2 //, in diam. ; singly or in zoogloea. They occur in various infusions and putrefying fluids in company with Bacterium termo. Micrococcus cumulatus tenuis (Besser). — Large cocci, oval ; in Colonies on agar have a brown nucleus. Inoculated in the depth of gela- tine they form a white filament, and on the surface a transparent layer. In broth there is an abundant deposit, and the supernatant liquid is clear. They occur in mucus from the nose. Micrococcus endocarditidis ru- gatus (Weichselbaum). — Cocci re- sembling pyogenic staphylococci. Colonies have a brown or yellow- ish-brown nucleus. In the depth of agar there is a slight growth in the track of the needle, and a wrinkled, waxy layer on the surface. On potato the growth is dry and brownish. On blood serum the growth is colourless and adherent. In jectedsubcutaneously, they pro- duce, in rabbits, local swelling and redness, and suppuration in guinea- pigs. Injected into the veins after injury to the aortic valves, they produce endocarditis. They were isolated from a case of ulcerative endocarditis. Micrococcus fervidosus (Ada- metz). — Cocci '6 /A in diam., in pairs and in masses. The deep colonies are pale-yellow, and look like watery droplets ; but superficial colonies are granular and irregular with jagged edges. Inoculated in the depth of gela- tine a granular filament develops in the track of the needle, and on the surface a circular patch with den- tated margin. On agar the growth is white and slimy, and on potato greyish - white. They occur in water. Micrococcus Finlayensis (Sternberg). — Cocci '5 to '1 p. in diam., singly, in pairs, tetrads, and in masses. In the depth of gelatine they produce a growth in the track of the needle, with liquefaction at the upper part with a pale-yellow deposit. On agar the growth is pale-yellow. They were isolated from the liver in a fatal case of yellow fever. Micrococcus flavus desidens (Fliigge). — Cocci singly, in pairs, or chains of a few elements. Colonies yellowish-white. Inoculated in the depth of gela- tine they grow along the track of 554 DESCRIPTION OF SPECIES. the needle, and form a yellowish- brown layer at the point of punc- ture. Later liquefaction sets in, and a deposit forms at the bottom of the turbid liquid. They occur in air and in water. Micrococcus flavus lique- faciens (Fliigge).— Cocci mostly in twos and threes, also in masses. Small yellow colonies appear after two or three days, which have a shallow depressed zone surrounding j them. Under a low power they are granular and yellowish-brown, with lines radiating from the centre to the circumference. Later they liquefy the gelatine, and coalesce. Inoculated in the depth of gelatine the cocci produce spherical yellow colonies in two days along the track of the needle. These become con- fluent, and at the end of eight days the whole of the jelly has become liquid ; later the upper part becomes clear, and a yellow mass subsides to the bottom of the tube. They occur in air and in water. Micrococcus flavus tardigra- dus (Fliigge). — Large cocci showing at times peculiar dark poles ; gener- ally arranged in masses. Colonies develop slowly ; the superficial ones have a smooth wax- like surface with projecting centre ; those below the surface are of a dark chrome-yellow colour, and are round or oval. Inoculated in gelatine the cocci develop slowly along the track of the needle, forming small isolated colonies ; the gelatine is not liquefied. They occur in air and in water. Micrococcus fcetidus(Klamann). , — Cocci singly, in pairs, and short j chains and masses. Colonies circular or oval, white. Inoculated in the depth of gela- | tine a pure white, shining growth j forms in concentric circles at the ! point of puncture, and develops a [ brownish colour ; and liquefaction occurs after a time, and extends along the needle track. A white layer spreads over the surface of agar. On potato the growth is slimy and grey in colour, with a red tinge. Cultures produce an odour like that of ozaena. They were isolated from the nose. Micrococcus ftetidus (Rosen- bach). — Small oval cocci. Cultivated in agar-agar they develop gas-bubbles and a foetid odour. They were isolated from carious teeth. Micrococcus Freudenreichi (Guillebeau). — Large cocci, singly and in chains. Colonies are granular and puncti- form. In broth turbidity is produced, and, later, a flocculent deposit. On potato a shining film develops, yellowish or brownish-yellow in colour. In milk the cultures become viscous, and can be drawn out into threads several yards in length. They were isolated from milk with viscous fermentation. Micrococcus fuscus (Maschek). — Cocci oval. Colonies pale-brown or black. Inoculated in the depth of gela- tine there is a slight growth along the track of the needle, and a brown layer forms on the surface followed by liquefaction. On potato the growth is brown or brownish-black and slimy. Cultures give off an odour of putrefaction. They occur in water. Micrococcus gingiyae pyogenes (Miller). — Large cocci, singly and in pairs. Colonies spherical, with sharp contours. Inoculated in the depth of gela- tine there is an abundant growth along the track of the needle and on the free surface. On agar a thick film develops, with a faint tinge of purple by transmitted light. Injected into mice subcutaneously they produce local suppuration, and sometimes death. Injected into the peritoneal cavity they produce peritonitis and death. DESCRIPTION OF SPECIES. 555 They were isolated from an abscess of the gums. Micrococcus gonorrhoeas (p. 190). Micrococcus havaniensis (Sternberg). — Cocci 4'5 \i in diam. The colonies are circular and of a blood-red colour. The cocci inoculated in the depth of gelatine produce a colourless growth in the track of the needle and a carmine patch on the surface. On agar and on potato they form a thick irregular carmine layer. Micrococcus in Biskra-button (Heydenreich). — Cocci in pairs, •86 to 1 p. in length, occasionally tetrads : capsulated. Inoculated in the depth of gela- tine they form a greyish-white fila- ment composed of closely packed colonies, and a yellowish- white film on the free surface. Liquefaction commences at the upper part of the needle track in a few days, forming a funnel which extends until, in two weeks, the gelatine is com- pletely liquefied. On the surface of agar a shining white or yellowish-white layer de- velops in twenty-four hours. On potato the growth is similar. Inoculations are said to produce in rabbits, dogs, fowls, sheep and horses a morbid condition of the skin similar to the disease known as Biskra-button or Pendjeh sore. In man they produce suppuration when rubbed on the skin. They were isolated from the disease known as Pendjeh sore, Biskra-button or clou de Btskra. Micrococcus in gangrenous mastitis in sheep.— Cocci singly, in pairs, and in masses. Colonies are spherical, white, and under a low power have a brown nucleus and transparent margin. The cocci inoculated in the depth of gelatine produce a conical area of liquefied jelly with a copious white deposit. On agar they produce a white layer, which later turns yellowish in colour. On potato they form a greyish growth. Injected into the mamary gland of sheep they produce inflammatory oedema, and a fatal result in twenty-four to forty-eight hours. In rabbits they are pyogenic. They were isolated from the milk in cases of gangrenous mastitis in sheep. Micrococcus in infectious p 1 e u r o-pneumonia (Poels and Nolen)— p. 242. Micrococcus in influenza (Fis- chel). — Cocci from 1 to 1-25 p in diam., singly, in pairs, and chains. Extremely minute colonies appear in three days. Inoculated in the depth of gela- tine a milk-white filament forms along the track of the needle. Lique- faction commences in four days at the upper part, and extends slowly. On agar the colonies are pure- white. On potato the growth is yellowish- white. They do not grow on blood serum or in milk. Intravenous injection in dogs is said to produce symptoms like distemper. They were obtained from the blood in cases of influenza. Micrococcus in influenza (Kircbner). — Cocci in pairs and chains ; capsulated. They grow at 37° C. The colonies are transparent, whitish. On the surface of agar there is an abundant growth, but it is limited in the depth of the jelly. Inoculation experiments were inconclusive. They were obtained from the sputum in cases of influenza. Micrococcus in pemphigus (Almquist). — Cocci '5 to 1 /i in diam., singly and in pairs ; identi- cal with Staphylococcus pyogenes aureus. • The cocci vaccinated on the arm are said to have produced bullae. They were isolated from pem- phigoid bullas in children. Micrococcus in pemphigus (Demme). — Cocci -8 to 1*4 p. in 556 DESCRIPTION OF SPECIES. diam., singly, in pairs, and in masses. They can be cultivated at 37° C. The colonies on agar are milk- white and prominent. Later, off- shoots occur from the margin, producing a resetted appearance. Inoculated in the depth of gelatine the cocci form clubbed or stalactitic out -growths from the filament which develops in the track of the needle. On the surface of agar a creamy layer is formed with similar off- shoots. Injected into the lungs of guinea- pigs they are said to produce broncho-pneumonia. They were obtained from the bullae in acute pemphigus. Micrococcus in pneumonia (Manfredi). — Oval cocci -6 to 1 /* in width, 1 to 1'5 p in length, singly, in pairs, and short chains. Colonies on gelatine are circular, whitish, and later spread out and become bluish by transmitted light, and of a pearly lustre by reflected light. Inoculated in the depth of gela- tine there is a limited growth along the track of the needle. On blood serum they form a shining, granular, faintly greenish- yellow layer. They also can be cultivated on potato and in broth. They are pathogenic in dogs, rabbits, guinea-pigs, mice and birds. Birds die in a few days ; mammals in from one to three weeks. After death new growths composed of granulation tissue are found in the internal organs, varying in size from a millet seed to a pea. They were obtained from the sputum of pneumonia complicating measles. Micrococcus in progressive abscess formation in rabbits (Koch).— Cocci only about '15 p. in diam., principally in thick zoogloea. The disease was induced by the in- jection into rabbits of decomposing blood. At the place of injection a spreading abscess formed, which was fatal to the animal in about twelve days. No bacteria were observed : in the blood, but in the walls of the abscess thick masses of cocci were found. The pus is infectious, causing the same disease in healthy rabbits. Micrococcus in pyaemia in rabbits (Koch). — Round cocci and diplococci "25 /u, in diam. The disease was produced by the subcutaneous injection, in a rabbit, of distilled water in which the skin of a mouse had been macerated. At the autopsy there were found great infiltration around the site of injection, peritonitis^ and accumu- lations in the liver and lungs ; in short, the appearances of pyaemia. FIG. 220.— MICKOCOCCUS IN PYAEMIA IN RABBITS : VESSEL FROM THE CORTEX OF THE KIDNEY x 700. a, Nuclei of the vascular wall ; c, i Masses of micrococci adherent to the wall and enclosing blood - corpuscles (Koch). In the capillaries of the organs examined, masses of cocci were observed enclosing blood-corpuscles. Fresh inoculations in rabbits with exudation-fluid, or blood from the heart, reproduced the same disease. Micrococcus in septicaemia in rabbits. — Ellipsoidal cocci -8 to 1 j IL in largest diam. The disease was produced by the injection of j putrid meat infusion. After death ' slight oedema was noted at the site of injection, slight extravasation of blood, and great enlargement of the spleen. No emboli or peri- tonitis resulted. Masses of cocci 1 were found in the capillaries of DESCRIPTION OF SPECIES. 557 different organs, especially in the glomeruli of the kidneys. Rabbits and mice inoculated with blood from the heart proved susceptible to the disease. Micrococcus in syphilis (Disse and Taguchi). — Cocci and diplo- cocci. They produce a greyish-white growth in nutrient media. They are said to produce inflam- matory changes in the internal organs and disease of the blood- vessels when inoculated in dogs, rabbits and sheep. They were obtained from the blood in cases of syphilis. Micrococcus in trachoma (p. 190). Micrococcus in yellow fever (p. 260). Micrococcus lactis viscosus (Conn). — Cocci in pairs and chains. Colonies circular and granular. Inoculated in the depth of gela- tine liquefaction begins at the upper part of the needle track, and extends until the gelatine is completely liquefied. The liquefied gelatine is viscous, and may be drawn out in long threads. On the surface of agar they form a white, shining layer. In broth there is an abundant growth, and a film on the surface. They coagulate milk, producing butyric acid, and giving it a bitter taste. They were obtained from bitter cream. Micrococcus luteus (Cohn).— Oval cocci 1 to 1'2 /* in diam. Colonies yellow, with irregular contours, and granular. Inoculated in the depth of gela- tine a granular filament develops in the track of the needle; and on the free surface a yellow patch. On agar the growth is slimy and yellow. On potato the growth is yellow, and after a time wrinkled. The pigment is insoluble in water, ether and alcohol, and not destroyed by acids or alkalies. They occur in water. Micrococcus luteus (Schrdter). — Cocci similar in size to the above, elliptical, with highly refractive cell contents. They form yellow drops of 1 to 3 mm. diam. on boiled potato ; and a thick, wrinkled, yellow skin on nutrient liquids. The colouring-matter is insoluble in water, and unchanged by sul- phuric acid or alkalies. Micrococcus ochroleucus (Prove). — Cocci *5 to '8 /M in diam., singly, in pairs, and short chains. Colonies minute and colourless, with crenated margin, from which, later, processes extend into the gela- tine, while the centre of the colony becomes yellow. On the surface of gelatine a film develops, which in a few days turns yellow. Old cultures have a pecu- liar smell. The yellow pigment can be ex- tracted with alcohol. It is insoluble in water, and decolorised by acids. They were obtained from human urine. Micrococcus of Forbes (p. 472). Micrococcus plumpsus (Brauti- gam). — Cocci *8 /z in diam., in masses. Colonies yellowish-white. Inoculated in the depth of gela- tine long delicate acicular processes shoot out from the needle track and on the free surface. On potato the growth is similar. They were isolated from water. Micrococcus pneumoniae crou- posae (p. 236). Micrococcus pyogenes tenuis (Rosen bach). — Cocci irregular, somewhat larger than staphylococci, and with much less tendency to form masses. The ends colour deeply, leaving a clear space in the middle. Inoculated in the depth of gela- tine a slightly opaque growth is formed. On agar a thin deposit appears along the needle track, which is almost as clear as glass. They occur in the pus of un- opened abscesses, but not often, as they were found by Rosenbach only in three out of thirty-nine cases. 558 DESCRIPTION OF SPECIES. Micrococcus rpsettaceus (Zim- mermann). — Cocci '7 to 1 p. in diam., singly, and in masses. Colonies circular, whitish, or greyish-yellow. Inoculated in the depth of gela- tine the growth is very scanty in the track of the needle, but spreads over the surface as a grey rosette. On agar a smooth layer with den- tated margin is formed. On potato the growth is faintly yellowish. They occur in water. Micrococcus roseus (Eisen- berg). — Cocci forming pink colonies, and a rose-coloured growth on the surface of nutrient agar-agar. They were found in sputum in a case of influenza. Micrococcus salivarius septi- CUS (Biondi). — Oval cocci, diplo- cocci, in chains and small masses ; capsulated. They grow best on acid gelatine, or in an atmosphere of carbonic acid. Colonies are small and circular, with an opalescent centre and a transparent margin, with sharply defined outline. In the interior of the colonies there is an appearance of a network. Inoculated in the depth of gela- tine the cocci form a delicate fila- ment and white dots on the free surface. On agar the cultures should be made direct from the blood. The growth appears on the surface and resembles dewdrops. Broth cultures remain clear. They are fatal to mice in twenty- four to seventy-two hours, and to rabbits in fifteen to thirty days, producing septicaemia. Attenuated cultures are said to give immunity. They were found in the saliva of .healthy and diseased persons. Micrococcus stellatus (Mas- chek). — Cocci singly. Colonies stellate. Inoculated in the depth of gela- tine a branching growth appears in the track of the needle. The jelly becomes brownish. On potato the growth is brownish- yellow and shining. They occur in water. Micrococcus tetragenus (Gaffkey). — Cocci about 1 /* in diam., in tetrads, and surrounded by a hyaline capsule. Colonies form in twenty-four to forty-eight hours as small white dots, which are finely granular, and have a vitreous lustre ; when they reach the surface they form thick raised masses. Inoculated in the depth of gela- tine the cocci form an irregular white growth, especially in the upper part of the track of the needle. On agar the colonies occur along the needle track, and are white, round and circumscribed. On potato they form a thick, slimy, viscous layer. White mice inoculated with a minute quantity of a pure-cultiva- tion die in from two to ten days, and the groups of the characteristic tetrads may be found in the capil- laries throughout the body, especi- ally in the spleen, lung and kidney. Double infection can be produced by inoculating a mouse with a pure cultivation of Bacillus an- thracis two or three days after inoculation with Micrococcus tetra- genus. On examination after death, the capillaries of the lungs, liver and kidney are filled with both anthrax bacilli and masses of tetrads (Plate V., Fig. 3). Micrococcus tetragenus mobilis ventriculi (Mendoza).— Cocci in tetrads ; capsulated ; motile. Colonies circular, whitish and granular. Inoculated in the depth of gela- tine they grow on the free surface only, and give off a peculiar odour. They were isolated from the stomach. Micrococcus tetragenus sub- flavus (Von Besser).— Cocci singly, and in tetrads. They do not grow on gelatine. Colonies on agar are brown and irregular in contour. DESCRIPTION OF SPECIES. 559 On the surface of agar the cocci form a greyish-white band, which turns brown at the periphery, and later is all dark or orange-yellow. On potato the growth is brown. They occur in nasal mucus. Micrococcus tetragenus ver- satilis (Sternberg and Finlay). — Cocci varying in size from '5 to 1*5 yn, in tetrads and irregular groups. Colonies are circular and lemon- yellow in colour. Inoculated in the depth of gela- tine there is very scanty develop- ment along the line of puncture, and the gelatine is liquefied in the form of a cup near the surface. At the bottom of the liquefied gelatine a viscid, pale-yellow mass accumulates. On the surface of agar a thick, viscid, yellow layer is formed along the line of inoculation, which gradu- ally extends over the entire sur- face. The colour varies from cream- yellow to lemon-yellow, and the surface is moist and shining. On potato there is a similar growth. They were isolated from the skin of patients suffering from yellow fever, from mosquitoes after attacking these patients, and from the air. Micrococcus ureae liquefaciens (Fliigge). — Cocci spherical, 1*25 to 2 fj, in diam., singly, or in chains of three to ten elements, or in irregular groups. Colonies appear in two days as small white points. They have sharply defined edges and a granular surface. The gelatine gradually liquefies, and the edges of the colo- nies become irregular. The cocci inoculated in the depth of gelatine produce a continuous white line along the track of the needle. Finally, the whole of the gelatine liquefies, and appears as a whitish-turbid fluid with a thick whitish-yellow deposit at the bottom. They were obtained from urine. Micro co ccusversicolor (Fliigge). — Small cocci, in pairs and in masses. White colonies develop in twenty- four hours ; after two days they are yellowish, with sharp contours of yellowish-green colour, and finely granular. The superficial colonies form flat deposits, 2'6 mm. in size, increasing to 10 mm. after four or five days. On the surface of gelatine the cocci form a shining layer with a greenish or bluish shimmer like mother-of-pearl . Inoculated in the depth of gela- tine the growth is composed of spherical yellowish colonies, and on the free surface they form an iridescent film. They occur in the air. Micrococcus violaceus (Schro- ter). — Cocci or elliptical cells, de- scribed as uniting into violet-blue gelatinous spots, which again unite to form larger patches. The colonies on gelatine are violet in colour. Inoculated in the depth of gela- tine the growth is scanty in the track of the needle. On the surface of gelatine they form a bluish-violet layer, and the same on agar and potato. They were observed on boiled potatoes exposed to the air, and are also found in water. Micrococcus viticulosus (Katz). — Oval cocci 1 // in width, and 1'2 /u, in length, in masses, but without formation of much gelatinous ma- terial. The superficial colonies are quite different in appearance from the deep colonies. From the deep colonies fine hairlike tendrils are thrown off from a centre, forming a very delicate and extensive net- work. The threads are found to consist of zooglcea masses, irregular in size, arranged like strings of beads. The colonies which are ex- posed to the air form a thin layer of muddy-white gelatinous sub- stance, which rapidly spreads, some- times sending out hairlike processes into the depth of the gelatine. Inoculated in the depth of gela- tine a delicate feather-like growth occurs along the track of the needle, 560 DESCRIPTION OF SPECIES. and on the free surface they pro- duce the appearance which has been described in colonies. This micro- organism is exceedingly rare. It was obtained from a contaminated culture. Monas Okenii. — Short cylindrical cells, 5 n wide, 8 to 15 p. long, with rounded ends. They exhibit lively movements, each end being provided with a flagellum twice as long as the cell itself. They have pale-red cell-substance, with dark grains. They occur in stagnant water. Monas vinosa.— Round or oval cells of about 2'5 p. in diam., often united in pairs. Their motion is slow and tremulous, and the cell- substance is pale-red with dark grains interspersed. Flagella have not been observed. They were found in water with decaying vegetable matter. Monas Warmingii.— Cylindrical cells, rounded at the ends, 15 //, long, 5 to 8 /j. broad. They are possessed of a flagellum at each end, and exhibit rapid, irregular movements. The cell-substance is pale-red, inter- spersed at the ends with dark-red grains. Myconostoc gregarium (Cohn). — The threads are very thin, colour- less, unarticulated, but fall apart into short cylindrical links when dried. They form gelatinous masses, 10 to 17 n in diam., singly or heaped into slimy drops 011 water in which algae are decomposing. Nitromonas of Winogradsky.— Very short rods, -9 to 1 p, in width, I'l to T8 p. in length. Singly, in masses, and in very short chains. They can be cultivated in silica- 'hey are active agents of nitrifi- cation. They were obtained from the soil. Pediococcus acidi lactici (Lindner). — Cocci '6 to 1 ^ in diam., singly, in pairs, and tetrads. Colonies colourless. On the surface of agar the cocci form a colourless layer. On potato the growth is almost invisible. The cocci produce lactic acid in solutions containing sugar. They occur in hay infusion and malt. Pediococcus cerevisiae (Balcke). — Cocci singly, in pairs, and tetrads. Colonies at first colourless, later yellowish-brown. Inoculated in the depth of gela- tine a greyish-white filament occurs in the track of the needle, and a white layer on the free surface. On agar the growth is transparent and iridescent, and on potato almost invisible. They were isolated from the air of a brewery. Pneuinobacillus liquefaciens bovis (p. 242). Proteus capsulatus septicus (Banti). — Rods isolated from a case of septica3mia, and identical with Proteus hominis capsulatus. Proteus hominis capsulatus (p. 224). Proteus in gangrene of the lung (Babes). — Rods -8 to 1-5 /* thick, irregular in form, and fila- ments with irregular enlargements. Colonies whitish and transparent, with ramifications extending over the gelatine. In the depth of gelatine a growth occurs along the track of the needle, and a ramifying growth on the free surface. On agar the growth is slightly yellowish. On potato the growth is brownish. They are extremely pathogenic in mice and guinea-pigs. They were isolated from a case of gangrene of the lung. Proteus microsepticus (Kar- linski). — Cocci, rods and filaments in morphology, and cultures re- sembling Proteus vulgaris. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle, forming a funnel with cloudy contents, and in a few days the whole, of the gelatine is liquid. They produce a general infection in mice, and death in twenty-four hours, and occasionally death in DESCRIFHON OF SPECIES. 561 rabbits, and local suppuration in guinea-pigs and white rats. They were isolated from pus in a fatal case of puerperal pyaemia. FIG. 221. — PROTEUS MIRABILIS : SWARMING ISLANDS ON THE SURFACE OF GELATINE, x 285 (HAUSER). Proteus mirabilis.— Cocci -4 /* to '9 /LI. They occur singly and in zoogloea, and sometimes in tetrads, pairs, chains, or as short rods in twos resembling Bacterium termo — concentric circles, which in time liquefies the medium. Similar movements are observed in capsule- cultivations as in Proteus vulgaris. They were isolated from putrid meat infusion. Proteus septic us (Babes).— Kods -4 /* in width, and filamentous forms. Colonies rapidly liquefy ^^s** the gelatine. Inoculated in the depth / of gelatine the bacilli form / a turbid funnel, or com- pletely liquefy the gelatine in twenty-four hours. On agar the growth is reticulated. On potato brownish - white. Cultures have an un- pleasant odour. They are pathogenic in mice. They were isolated from the organs in a case of human septicaemia. Proteus sulfureus (Lin- denborn). — Rods -8 /* in width, varying in length, and long filaments. They correspend in mor- phology and cultures with Proteus vulgaris. They produce sulphuretted hy- drogen in cultures. They were isolated from water. Proteus vulgaris (Hauser).— *%•» £ -i-^^sf* * » x *<7^ % ""^ Si /J/ ^« ^o/t \' £>.*flPvT.^ FIG. 222.— PROTEUS MIRABILIS.: INVOLUTION FORMS, x 524 (HAUSER). in fact, in all conceivable transition forms. Cultivated on nutrient gelatine they form a thick whitish layer in Rods varying in size ; some mea- sure 4 /n in length, and are almost as broad as long, and others vary from -94 to 1'25 p, long and '42 to 36 562 DESCRIPTION OF SPECIES. •63 fj. wide. They are actively motile. Cultivated on nutrient gelatine they convert it into a turbid, grey- ish-white liquid. If cultivated in a capsule containing 5 per cent, of nutrient gelatine, a few hours after inoculation the most characteristic movements of the individual bacilli FIG. 223. — PROTEUS VULGARIS, FROM SURFACE OF NUTRIENT GELATINE, x (HAUSER). are observed on the surface of the nutrient gelatine, although at this early stage no superficial liquefac- tion can be detected. Probably the movements depend upon the exist- ence of a thin liquid layer, as they are not observed if the nutrient medium contains 10 per cent, of gelatine. They were isolated from putrid meat infusion. Proteus Zenkeri.— Cocci -4 /*, in twos like Bacterium termo, and short rods 1 -65 /* long. Cultivated on nutrient gelatine no liquefaction results, but a thick whitish-grey layer is formed. The bacilli are motile, and the same phenomena are observed on the solid medium as in Proteus vulgaris. In cover-glass impressions most varied groupings of the bacilli are seen, and also developmental and involution-forms. They were isolated from putrid meat infusion. Pseudo-diphtheritic bacil- lus (p. 335). Pseudo-diplococcus pneu- nioniae (Bonome).— Oval cocci in pairs and short chains ; cap- sulated. Inoculated in the depth of gelatine small colonies develop in the track of the needle in twenty-four hours. On agar there is a scanty, moist growth. On potato an almost in- visible film. In broth the cocci grow rapidly, and the cultures give off a peculiar odour. They produce septicaemia in mice, guinea-pigs and rabbits. This micro-organism is probably a variety of the pneumococcus. They were isolated from a fatal case of cerebro-spinal meningitis. Rhabdomonas rosea. — Spindle-form rods, 3'8 to 5 p THE broad, 20 to 30 ^ long. They 3 exhibit slow, trembling move- ments, having at each end of the cell a flagellum. The cell-substance is very pale, with dark grains interspersed. They occur in brackish water. S a r c i n a alba. — Small cocci. They form small white colonies on nutrient gelatine. Inoculated in the depth of gela- tine they grow slightly along the needle track, but are heaped up on the surface without liquefying the gelatine. They are present in the air. S a r c i n a aurantiaca.— Cocci singly, in pairs, in tetrads, and in packets. Colonies orange-yellow. Inoculated in the depth i of DESCRIPTION OF SPECIES. 563 gelatine they slowly liquefy it along the whole needle track, and form on the surface an orange-yellow growth. On potatoes they slowly develop the same pigment. Sarcina Candida (Reinke).— Cocci 1-5 to 1-7 /x in diam., singly, in pairs, and in tetrads. Colonies are circular and shining, white, and later yellowish. Inoculated in the depth of gela- tine liquefaction quickly takes place along the track of the needle. On the surface of agar a white, moist layer develops. They were found in the air of breweries. Sarcina flava(De Bary).— Small cocci in packets. Inoculated in the depth of gela- tine they produce liquefaction. On agar they form a yellow layer. On potato the growth is limited and yellow. They were isolated from beer. Sarcina hyalina (Kiitzing). — Cocci round, 2'5 ^ in diam., almost colourless. United in families of 4 to 24 cells, reaching 15 /u in diam. They occur in marshes. Sarcina intestinalis (Zopf).— Cocci in groups of four or eight. Yery regular in form ; never in the large packets which occur in Sarcina ventriculi. They are found in the intestinal canal, especially the caecum, of poultry, particularly fowls and turkeys. Sarcina litoralis (Oersted).— Cocci 1-2 to 2 fj. in diam., bound together in 4 to 8 families, which, in their turn, may unite and in- clude as many as 64 tetrads. Plasma colourless ; in each cell 1 to 4 sulphur granules. They were found in sea water containing putrefying matter. Sarcina lutea (Schroter). — Cocci singly, in pairs, tetrads and packets. A single individual in a tetrad may be divided into two, or into four, so that a tetrad within a tetrad results. Colonies are round, slightly granular in appearance, and yellow. Inoculated in the depth of gelatine they grow rapidly ; the gelatine becomes liquefied, and the yellow growth sinks to the bottom of the tube. as* FIG. 224.— SARCINA x 600 (FLUGGE. Cultivated in agar they form a colourless growth along the track of the needle, and a bright canary- yellow layer upon the surface. On potato they form a yellow layer. They are present in air. Sarcina mobilis (Maurea).— Cocci 1-5 p. in diam., in pairs, and in tetrads. They are motile. Colonies, at first white, become brick-red. Inoculated in the depth of gela- tine, there is, after several days, a slight growth along the track of the needle, and a patch of growth on the free surface which gradually turns red. In about two weeks liquefaction produces a funnel- shaped appearance ; later the lique- faction extends to the sides of the test-tube. In broth turbidity is produced, and a yellowish-red deposit. On agar the growth, at first white, changes to a brick-red colour. There is no growth on potato, and milk is not coagulated. They were isolated from ascitic fluid. Sarcina pulmonum (Hauser).— Cocci from 1 to 1*5 ft in diam., in tetrads and packets. Colonies white and small. They are coarsely granular. Inoculated in the depth of gela- tine the growth is scanty in the track of the needle, but on the free surface there is a circular, well- defined, translucent patch, which 564 DESCRIPTION OF SPECIES. later becomes greyish -brown, shin- ing, wrinkled and irregular. On potato the growth is very slight and limited. They cause ammoniacal decom- position of urine. They were isolated from phthisi- cal sputum. Sarcina Reitenbachii (Caspary). — Cocci about 1'5 to 2 '5 \i in diam., at the time of division lengthened to 4 \i. Mostly united together from 4 to 8 in number ; occasion- ally 16 or more. Colourless cell- wall, lined with rose -red layer of plasma. They were found on rotting water-plants. Sarcina rosea (Schroter). — Large cocci, in packets. Inoculated in the depth of gela- tine, liquefaction quickly takes place, and cultures after a time have a reddish colour. On agar the growth is slow and limited. On potato the growth is abundant and of a bright-red colour. In broth they produce turbidity, and a red deposit. They occur in the air. Sarcina urinse (Welcker).— Very small cocci, 1'2 /z in diam., united in families of 8 to 64. They were observed in urine. Sarcina yentriculi (Goodsir).— Cocci reaching 4 /* in diam., united in groups of four, or multiples of four, producing cubes or packets with rounded-off corners. Contents of the cells are greenish or yellow- ish-red. Colonies are round and yellowish. A yellowish growth forms on the surface of oblique gelatine without liquefaction. On potato they form a yellow growth, and on serum also. They grow well in hay-infusion, forming brownish scales and a simi- larly-coloured deposit. They occur in the stomach of man and animals in health and disease, and were first detected in vomit. Sphaerotilus natans.— Cells 4 to 9 /z long, and 3 /x thick, united in a gelatinous sheath to form threads. The cells comprise rods and cocci- forms ; the cocci are set free, and develop into rods, which again form threads. In the last a false branch- ing has been observed. The plasma of the cells break up into minute, strongly-refractive portions, which develop into round spores, at first of a red and afterwards a brown colour. They occur in stagnant and flow- ing water contaminated with or- ganic matter, and form floating flakes of a white, yellow, rust-red, or yellow-brown colour. Spirillum amyliferum (Van Tieghem). — Filaments 6ju in length and 1'4 to 1'5 /* in width ; with from 2 to 4 screw curves. They act as a strong ferment in the absence of air. They occur in water. Spirillum anserum (Sakharoff). — Spirilla resembling the Spiro- chae ta 0 bermeieri. Extremely motile. They have not been cultivated artificially. Blood from diseased geese, con- taining the spirilla, produces the disease when inoculated in healthy birds. The geese suffer from diarrhoea, and die in about a week. They were found in the blood of geese suffering from an epidemic form of septicaBmia prevailing at some of the stations on the Trans- caucasian Railway. Spirillum attenuatum (Warm- ing).— Threads much attenuated at the ends, which consist usually of three spirals. The middle spiral is about 11 p, high and 6 p. in diam., and the end ones 10 ft high and 2 p. in diam. They are found in brackish water. Spirillum aureum (Weibel). — Curved rods with blunt ends, spi- rilla and spirilliform filaments, and involution forms. Colonies are circular and golden- yellow. Inoculated in the depth of gela- tine, a finely granular growth forms in the track of the needle, and a yellow-ochre prominent mass on the free surface. DESCRIFHON OF SPECIES. 565 On agar a greyish growth ex- tends over the surface, and later prominent yellowish heaps make their appearance. On potato there is an abundant growth of a golden-yellow colour. They were found in sewage mud. Spirillum cholerae Asiaticse (p. 301). Spirillum choleroides (Buj wid). — Curved rods very similar mor- phologically and in cultures to Koch's comma-bacilli. They were isolated from river water. Spirillum choleroides (Orlow- sld). — Curved rods very similar to Koch's comma-bacilli. They were found in well water. Spirillum concentricum (Kitasato). — Short spirilla, and spirilliform filaments. Colonies are circular, and com- posed of concentric rings alternately opaque and transparent. Inoculated in the depth of gela- tine there is only a little growth in the track of the needle, and a cloudy growth on the surface extending into the jelly. On agar the growth is extremely adherent. In broth they produce turbidity, which disappears after a time ; and there is a slimy deposit at the bottom of the tube. They were found in putrefying blood. Spirillum dentium (Miller).— Spirals 10 to 20 p, in length, pointed at the ends. They have not been cultivated. They occur in the deposit on the teeth, and in company with Lepto- thrix buccalis in carious teeth. Spirillum flavescens (Weibel). — Commas thicker than those found in Asiatic cholera, spirilla, and spirilliform filaments. Colonies yellowish. Inoculated in the depth of gela- tine a finely granular filament develops in the track of the needle, and on the free surface a pale- yellow patch. On agar the growth, at first greyish- white, becomes yellow, and forms a thick layer. On potato the growth is abun- dant, and similar in colour. They were found in sewage mud. Spirillum flavum (Weibel).— Spirilla morphologically identical with spirillum aureum. Colonies on gelatine are pale- yellow, and later the colour is more intense. On agar and potato they form a layer the colour of yellow-ochre. They were isolated from sewage mud. Spirillum leucomelaneum (Koch). — Dark and glass-like spaces alternate in the spirillum, resulting from a regular arrangement of the dark granular contents. A rare form observed in water covering rotting algaB. Spirillum linguae (Weibel). — Curved rods, spirilla, and spirilli- form filaments, and involution forms. Colonies are composed of inter- lacing filaments, and offshoots extend into the surrounding gela- tine. Inoculated in the depth of gela- tine a delicate growth occurs in the needle track. On agar the growth is whitish and granular. In broth a cloudiness is produced, as well as a flocculent deposit. They are especially distinguished from other spirilla described by Weibel by their staining by Gram's method. They were isolated from the tongue. Spirillum marinum (Russell). — Curved rods, and spiral filaments. Colonies circular, granular and striated ; later, flocculent masses float in liquefied areas. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle, and a mem- brane forms on the surface of the cloudy liquid. On agar the growth is yellowish and abundant. On potato a thick, waxy mass 366 DESCRIPTION OF SPECIES. develops, and extends over the surface. Broth made with sea- water be- comes rapidly turbid. They were obtained from sea- water. Spirillum Metchnikovi (p. 373). Spirillum nasale (Weibel).— Large curved rods and spirilliform filaments. Non-motile. Colonies circular, finely granular, and brownish-yellow. Inoculated in the depth of gela- tine a delicate growth develops in the track of the needle. On the surface of agar they form a whitish slimy film. They occur in nasal mucus. Spirillum Obermeieri (p. 258). Spirillum of Finkler and Prior (p. 258). Spirillum of Gimther (Vibrio aqudtilis) . — Curved rods, very simi- lar to Koch's comma-bacilli, but there is no growth on potato. They occur in water. Spirillum of Miller.— Curved rods, singly, in pairs, and spiral filaments. They liquefy gelatine. They were isolated from carious teeth. Spirillum of Neisser (Vibrio berolinensis). — Similar to Koch's comma-bacillus, but smaller. Colonies colourless, granular and transparent, liquefying the gelatine much more slowly than Koch's comma-bacillus. Inoculated in the depth of gela- tine they produce a growth similar to that of Koch's comma-bacilli, but much more slowly in milk. In broth they grow abundantly. They were found in water. Spirillum of Renon.— Curved rods longer and broader than the comma-bacilli of Koch. Colonies yellowish, with dark nucleus. In gelatine the cultures resemble those of Koch's comma-bacillus. On agar the growth is white and abundant. They cause turbidity in broth. They were isolated from impure water from a well. Spirillum of Smith (vide Spirillum suis). Spirillum of Weib-el.— Curved rods resembling Koch's comma- bacilli, morphologically, and in cultures on jelly. The gelatine is more quickly liquefied. There is no growth on potato. They occur in well water. Spirillum plicatile (Ehrenberg). — Thin threads 2'25 fj, in breadth, 110 to 125 /u, long, occurring also in spirulinar forms. The threads have primary and secondary windings ; the former are in each example of equal size, but the latter are often FIG 225.— SPIKOCH.ETA PLICATILE. irregular. Their ends are cut off bluntly, and they exhibit rapid movement. They occur abundantly in marsh- water in summer, and can be ob- tained by allowing algae to decom- pose in water. On cultivation the threads break up into long rods, short rods, and, finally, cocci. This change is rendered visible by making cover-glass preparations, and stain- ing with aniline dyes. Spirillum rosaceum (Klein). — Resembles Spirillum undula, but is reddish in colour ; the colouring- matter is insoluble in water, alcohol or chloroform. Spirillum Ros enbergii.— Threads with 1 to 1£ windings, 4 DESCRIPTION OF SPECIES. 567 to 12 p long, 1-5 to 2-6 p thick. They are colourless, but the con- tents include strongly refractive sulphur granules. Also spirals 6 to 7'5 \i in height, which are actively motile. They were found in brackish water. Spirillum rubrum (Esmarck). — Curved rods, spirilla and spiro- chetae. They are actively motile. The growth on artificial media is extremely slow. Inoculated in the depth of gela- tine they grow along the track of the needle, forming a filament of a wine-red colour, without causing liquefaction ; and on the free sur- face the growth is colourless. In broth long spirillar threads are formed. They were isolated from the putrid tissues of a mouse. Spirillum rufum (Perty).— Filaments from 8 to 16 /z in length, with 1^ to 4 screw curves ; non- segmented ; chiefly motile ; tinged with red. They form rose or dark red spots on the sides of wells. Spirillum sanguineum (Cohn). — Threads 3 \L and more in thick- ness, with 2 to 2£ spirals, each 9 to 12 /u. high. The ends are provided with flagella. Their colour is due to the presence of reddish granules contained in the cells. They were observed in brackish water containing putrefying sub- stances. (Vide Beggiatoa roseo- persicina. ) Spirillum saprophiles. — (I.) Curved rods with pointed ends, -6 /L* in width, 3 \JL in length ; spirilla, spirilliform filaments, and involu- tion forms. Colonies yellowish or greenish- yellow. Inoculated in the depth of gela- tine a white growth forms in the track of the needle, later becoming yellowish ; and on the free surface there is a white growth, and beyond this a transparent film spreads over the jelly. On agar the growthi s creamy, and the jelly clouded beneath it. On potato the growth is slimy and yellowish or dark-brown in colour. They were obtained from sewage FIG. 226.— COMMA-BACILLI IN WATER CONTAMINATED WITH SEWAGE. mud and decomposing hay infu- sion. (II.) Curved rods about 2 p in length, with blunt ends and in pairs. Extremely motile. Colonies circular and yellowish- brown. Inoculated in the depth of gela- tine a white growth develops in the track of the needle, and later becomes yellowish-red ; on the free surface a white patch forms, sur- rounded by a transparent film. In the depth of agar there is no growth in the track of the needle, but a yellowish-white patch on the free surface adherent to the jelly. On potato the growth is also adherent, and in appearance shining and brownish-green. They were isolated from decom- posing hay infusion. (III.) Curved rods, spirilla, and spirilliform filaments, and involu- tion forms. Colonies are circular, granular, and with irregular margin ; yellow in the centre, and white at the periphery. Inoculated in the depth of gela- tine a white growth develops in the track of the needle, and on the surface, without producing lique- faction. On the surface of agar the growth is white. On potato the growth is distinctly brown. 568 DESCRIPTION OF SPECIES. They were isolated from sewage mud. Spirillum serpens (Miiller).— Long spirilliform filaments ; often collected in masses. They were observed in vegetable infusions and stagnant water. Spirillum sputigenum (Lewis). — Curved rods, very similar to the comma-bacilli of Koch ; but many observers having failed in repeated ,>\vvVv» <&?.<&• FIG. 227. — COMMA-BACILLI OP THE MOUTH, x 700 (VAN ERMENGEM). attempts to cultivate them, main- tain that they are biologically dis- tinct from those associated with Asiatic cholera. Klein asserts that they can be cultivated in an acid gelatine, and that they are iden- tical with Koch's comma-bacilli in their mode of growth. They occur with other bacteria in saliva and in scrapings from carious teeth. Spirillum suis (Smith).— Com- mas and spirilla. Colonies in gelatine are circular, granular and brownish, and later appear to be composed of concen- tric rings. The gelatine is not liquefied. Broth with 1 per cent, of peptone becomes in a few days clouded. On potato they develop a thin yellowish layer. The commas are said to be slightly larger than those obtained from Asiatic cholera, and are not pathogenic. They were obtained from the large intestine in swine. Spirillum tenue.— Very thin threads, with at least 1J, usually 2 to 5 spirals. Height of a single screw is 2 to 3 /u, and the length of spiral therefore 4 to 15 p.. They are very swiftly motile. They often occur in dense felted swarms in vegetable infusions. Spirillum tyrogenum (Deneke). — Curved rods, slightly smaller than Koch's comma-bacilli, with a great tendency to form long spirillar threads (Fig. 228). FIG. 228. — DENEKE'S COMMA-BACILLI FROM CHEESE, x 700 '.F Colonies on plate-cultivations are sharply defined and of a greenish-brown colour. After a time they liquefy the gelatine, but the liquefaction is much more marked than in colonies of Koch's commas of the same age, though not so rapid as in the case of the commas of cholera nostras. Inoculated in the depth of nu- trient gelatine a turbid liquefaction occurs along the needle track, and on the surface of nutrient agar-agar a yellowish-white layer develops. Inoculation of potatoes gives no result. Administration of the bacilli by the mouth, in the manner employed for testing the pathogenic effect of Koch's bacilli, produced a fatal result in a few cases ; on the other hand, injection into the duodenum failed entirely. The pathogenic properties may be therefore con- sidered as not yet established. They were isolated from old cheese. Spirillum undula.— Threads 1-1 to 1-4 fi thick, 9 to 12 ^ long ; spirals 4'5 fi high ; each thread has 1| to 3 spirals. They are actively motile, and possess a flagellum at each end. They occur in various infusions. Spirillum vulutans(Ehrenberg). —Threads 1-5 to 2 p, thick, 25 to 30 p long, tapering towards their DESCRIPTION OF SPECIES. 569 extremities, which are rounded off. They possess dark granular con- tents. Each thread has 2£ to 3£ windings or spirals, whose height is 1» to 13 p. They have a flagellum at each end, and are sometimes motile, sometimes not. They are found in the water of marshes and in various infusions. Spirompnas Cohnii.— Colourless cells, consisting of 1^ spirals, with both ends acutely pointed and pro- vided with a flagellum. Breadth of the cells, 1-2 to 4 /u. They occur in water containing decomposing matter. Spiromonas volubilis (Perty). — Colourless, transparent cells, 15 to 18 p long. Rapidly motile, and revolving round a longitudinal axis. They occur in marsh-water and putrefying infusions. Staphylococcus pyogenes albus (p. 178). Staphylococcus pyogenes aureus (p. 176). Staphylococcus pyogenes cit- reus (p. 178). Staphylococcus pyosepticus (Heucourt and Richet). — Cocci identical with Staphylococcus pyo- genes aureus. Subcutaneous injection causes in rabbits intense oedema, and death in twenty-four hours. They were isolated from pus from an abscess in a dog. Staphylococcus salivarius pyo- genes (Biondi). — Cocci '3 to -5 /* in diam., singly and in masses. Colonies white and opalescent, producing liquefaction. Inoculated in the depth of gela- tine the growth appears in the track of the needle, and is followed by liquefaction. On agar the growth is orange- yellow. The cocci produce local suppura- tion when inoculated in animals. They were isolated from an abscess in a guinea-pig following subcutaneous injection of saliva. This coccus is probably identical with Staphylococcus pyogenes aureus. Staphylococcus viridis flaves- cens (Guttmann). — Cocci singly, in pairs and masses ; morphologically agreeing with Staphylococcus pyo- genes aureus. Colonies are greenish-yellow. Inoculated in the depth of gela- tine a filament forms composed of greyish colonies. On agar the growth is greenish- yellow. They grow well on potato. They were isolated from the vesicles of chicken-pox. Streptococcus acidi lactici (Grotenfeld). — Oval cocci -5 to 1 /n long, '3 to '6 //, in width, and long chains. They are partially anaerobic. Colonies are circular and white. Inoculated in the depth of gela- tine a growth occurs only in the track of the needle. Milk is coagulated. They were isolated from coagu- lated milk. Streptococcus albus (Tils).— Cocci forming motile chains. Colonies are flat and circular, with white periphery and dark nucleus, rapidly liquefying. Inoculated in the depth of gela- tine there is rapid liquefaction in the track of the needle, and a white deposit. On potato they form a white slimy layer. They were found in water. Streptococcus bombycis (p. 472). Streptococcus brevis (Lingels- heim). — Cocci singly, in pairs and chains, of eight to ten elements. Colonies on gelatine are circular and very minute. Inoculated in the depth of gela- tine there is a funnel-shaped cavity near the surface, and below this, in the track of the needle, small isolated colonies. On agar a yellowish-grey film develops along the line of inocula- tion. On potato there is a copious white growth in forty-eight hours. Broth is made turbid. They were isolated from healthy saliva. 570 DESCRIPTION OF SPECIES. Streptococcus cadaveris (Stern- berg).— The description corresponds with that of Streptococcus pyo- genes. Inoculated in the depth of gela- tine the colonies are said to be larger and more opaque. On the surface of agar they form a thin translucent layer. In broth little flocculi develop, composed of chains in which in some cases the elements varied considerably in size. They were obtained from the liver in a fatal case of yellow fever. Streptococcus coli gracilis (Escherich). — Cocci from -2 to '4 p, in diam., forming chains composed of from six to twenty elements. Some elements in a chain are irre- gular in form, and show transverse fission. The colonies are spherical and sink down in the liquefied gela- tine. Inoculated in the depth of gela- tine liquefaction occurs in the track of the needle on the second day, and a white deposit forms at the bottom of the liquid. In about a week the gelatine is completely liquefied. On agar there is a very slight growth. On blood-serum small scales develop. On potato the growth is com- posed of small white prominent colonies. Milk is coagulated. They were found in the evacua- tions of healthy infants. Streptococcus conglomeratus (Kurth). — Cocci and chains, identi- cal with Streptococcus pyogenes. They form an adherent film at the bottom of the tube, which is not broken up by agitation. This is observed in other varieties of Streptococcus pyogenes, and is not sufficient to distinguish it. They are pathogenic in mice. They were isolated from cases of scarlet fever. Streptococcus flavus desidens (Fliigge).— Cocci, diplococci, and short chains. They form yellowish- white colonies, which gradually sink down in the gelatine. Inoculated in the depth of gela- tine the cocci form china-white, confluent masses in the track of the needle, and on the surface a yellowish-brown slimy layer. They occur in air and in water, and were originally isolated from contaminated cultures. Streptococcus giganteus urethras (Lustgarten).— Cocci -8 to 1 p. in diam., forming chains composed of several hundred elements. In description they correspond with Streptococcus pyogenes. They do not grow at the tem- perature of the room. Colonies on agar are transparent and iridescent. They were isolated from the healthy urethra. Streptococcus Havaniensis (Sternberg). — Cocci '6 to -9 /u in diam., forming long chains, com- posed of cocci, in pairs, and oval elements showing transverse divi- sion. This streptococcus is probably a variety of Streptococcus pyo- genes. They were found in the acid vomit of a yellow-fever patient. Streptococcus in contagious mammitis in cows (Nocard and Mollereau). — Cocci spherical or oval, united in long chains. Colonies are spherical, granular, pale-yellow, or brownish by trans- mitted light. The cocci inoculated in the depth of gelatine produce a granular filament in the track of the needle. On the surface of nutrient gela- tine minute spherical colonies are formed, which are bluish by re- flected light. Injected into the mammary gland of cows and goats they produce mastitis. They were isolated from the niilk of cows suffering from contagious mammitis. From the description this strepto- coccus appears to be closely related DESCRIPTION OF SPECIES. 571 to, if not identical with, Strepto- coccus pyogenes bovis(Crookshank). Streptococcus in progressive tissue necrosis in mice.— Koch produced a disease in mice by sub- cutaneous injection of putrid blood. In tissue sections a chain coccus was found which was similar to Streptococcus pyogenes. FIG." 229.— STREPTOCOCCUS IN PRO- GRESSIVE TISSUE NECROSIS IN MICE. a, Necrotic cartilage cells, and (b) chains in masses ; c, isolated chains. (Koch.) Streptococcus in Strangles (Schutz). — Cocci forming long chains, which, it is said, do not grow on nutrient gelatine or agar, but form a transparent iridescent cul- ture on blood serum. Cultures in broth produced the disease in horses and mice. Rabbits, guinea-pigs, and pigeons are not affected. Strangles is a disease of the horse, associated with suppuration of the glands of the head and neck, prin- cipally in the sub-maxillary, sub- parotideal, and retro-pharyngeal regions. Schutz found that the pus contains streptococci and produces a fatal disease in mice. Streptococcus liquefaciens (Sternberg). — Spherical and oval cocci, *4 to '6 /z in diam., singly, in pairs and short chains. Inoculated in the depth of gela- tine liquefaction occurs rapidly in the track of the needle, and in a week the gelatine is completely liquefied, slightly opalescent, and a scanty deposit forms at the bottom of the tube. In the depth of agar a filament is formed composed of closely- crowded colonies. On potato a thin and limited dry white layer is formed along the line of inoculation in four to five days. They are non-pathogenic. They were isolated from the liver and intestines of fatal cases of yellow fever. Streptococcus mirabilis (Ros- coe and Lunt). — Cocci 4 p in diam., singly, and in long chains. The growth on nutrient media is very scanty. In broth the growth is composed of a mass of delicate filaments which collect at the bottom of the liquid. They were isolated from sewage. Streptococcus of Bonome.— Cocci forming chains. They corre- spond in description with Strepto- coccus pyogenes, but, it is said, they do not grow in gelatine or on blood-serum, and they are said to be distinguished by the characters of the colonies on agar plates. They are pathogenic. Inoculated in rabbits and white mice they produce symptoms similar to those produced by in- oculations of the pneumococcus. Sub-cultures rapidly lose their virulence. They were isolated from cases of cerebro-spinal meningitis. Streptococcus of Manneberg — Cocci -9 \i in diam., singly, in pairs, and in chains. Inoculated in the depth of gela- tine a white filament forms along the track of the needle composed of minute colonies. In about a month the filament is replaced by a funnel of semi-liquefied gelatine. On the surface of agar the 572 DESCRIPTION OF SPECIES. growth resembles Streptococcus pyogenes. "On potato they form a slimy layer. Milk is rapidly coagulated. They are pyogenic in dogs and rabbits. Injected into the veins they produce inflammation of the kidneys. They were isolated from the urine in a case of Bright' s disease. Streptococcus perniciosus psittacorum {Parrot disease). — Cocci, singly, in chains, and in zooglcea, have been described in connection with a disease of the grey parrot (Psittacus erithacus). This disease is fatal to about 80 per cent, of these parrots imported to Europe. They suffer from diarrhoea and general weakness ; their feathers are ruffled ; their wings hang loosely, and their eyelids close ; convulsions set in, and death fol- lows. At the autopsy greyish nodules are found in the lungs, liver, spleen and kidney. In and around the capillaries of these nodules, and in the blood of the heart, the cocci are found in great numbers in zooglcea, and more rarely in chains. Inflammatory change in the sur- rounding tissue is absent. Streptococcus pyogenes (p. 178). Streptococcus radiatus (Fliigge). — Cocci less than 1 ^ in diam., singly, in small masses, and sometimes in short chains. Colonies appear in twenty-four hours. They are white, with a yellowish-green sheen ; later they liquefy the gelatine and develop a circlet of rays. Inoculated in gelatine, isolated centres form along the track of the needle which throw out horizontal rays. At the same time a funnel- shaped area of liquefaction forms very slowly in the upper part. On potato the growth is yel- lowish-brown. They occur in air and in water. Streptococcus septicus (Fliigge). — Cocci in chains, indistinguishable microscopically from Streptococcus pyogenes. Colonies on gelatine grow more slowly than those of most strepto- cocci. They are pathogenic. Mice die in forty-eight to seventy-two hours after subcutaneous inoculation of a minute quantity of a cultivation. During the last twenty-four hours there is a distinct motor and sensory paralysis of the hind legs. In rabbits inoculation of the ear pro- duces local redness, then a general disease, and death in two or three days. They were found by Nicolaier,and independently by Guarneri, in earth. Streptococcus septicus lique- faciens (Babes).— Cocci -3 to -4 ^ in pairs, in short chains. Inoculated in the depth of gela- tine a granular filament forms in twenty-four hours along the track of the needle, followed by lique- faction of the gelatine forming a funnel in which the gelatine is clouded ; flat, whitish deposits form on the side of the funnel. On the surface of agar minute shining, transparent colonies are formed. On blood-serum the growth is almost invisible. They are pathogenic. Subcu- taneous injection in mice and rabbits produces local inflammation and oedema, followed by death in about a week. They were found in the blood and organs of a child which had died of septicaemia complicating scarlet fever. Streptococcus yermiformis (Tils). — Cocci forming motile chains. Colonies are yellowish-white, the central portion finely granular, the periphery radiated. Inoculated in the depth of gela- tine there is rapid liquefaction, and a yellowish deposit at the bottom of the liquid. On potato the culture forms a dirty-yellow layer. They were found in water. Streptothrix actinomycotica (p. 431). Streptothrix alba (Gasparini). — A variety of Actinomyces bovis. DESCRIPTION OF SPECIES. 573 Streptothrix asteroides { Ooxpora asteroides, Sauvageau and Radais ; Cladothrix asteroides, Eppinger). — Branching filaments which form on the surface of grape-sugar-agar a whitish growth, which is later of a brownish-yellow colour. Broth remains clear, and small pellicles float on the surface re- sembling drops of stearin. On potato they form snow-white points, which turn brick-red in colour, and are later covered with a delicate white efflorescence. The streptothrix is pathogenic in rabbits and guinea-pigs. It was isolated from pus. Streptothrix aurantiaca (Oospora aurantiaca, Sauvageau and Radais, and Doria). — Similar to Streptothrix asteroides. Streptothrix carnea (Doria; Oospora carnea, Sauvageau and Radais). — Similar to Streptothrix asteroides, but the cultures on gelatine are pink. They are not pathogenic. Streptothrix chromogenes (Gasparini ; Oospora chromogenes, Lehmann and Neumann). — Culti- vated on the surface of gelatine the filaments produce a chalky growth, and the jelly is coloured brown, and is slowly liquefied. i On potato the growth is yellowish or brown, and the potato itself is coloured dark brown or black. The streptothrix has been isolated from air and water and the con- tents of the stomach. Streptothrix farcinica (Bacille du farcin de bceuf, Nocard ; Oospora facinica, Sauvageau and Radais). Inoculated on the surface of gelatine there is in about two weeks a very scanty granular growth. In broth greyish pellicles develop with a dusty surface. They are pathogenic in cattle, guinea-pigs, and sheep. They were isolated from the disease known as farcin de bceuf. Streptothrix Forsteri (Cohn). — Cocci rods, and leptothrix threads. The threads are twisted in irregular spirals, and branch sparingly and irregularly. Screw-forms are pro- duced by the threads breaking up into small pieces. Colonies slowly liquefy gelatine. On agar they form a whitish growth. In broth they form shining masses, floating in clear liquid. They occur in the lachrymal canals of the human eye, in the form of closely felted masses, and in the air, and in fresh- and sea- water. Streptothrix Hofmanni (Micro- myces Hoffmanni, Gruber ; Oospora Hoffmanni, Sauvageau and Radais). —The filaments flourish in the ordin- ary culture media with the addition of sugar, but they do not grow on potato. They produce suppuration in rabbits. They were isolated from the air. Streptothrix liquefaciens (Cladothrix liquefaciens, Garten). — A variety of Actinomyces bovis. Streptothrix madurae (p. 449). Streptothrix musculorum SUis (Actinomyces suis, Bunker). — A variety of actinomyces found in the muscles of swine. Streptothrix odorif era (Oospora odor if era, Rullmann). Probably identical with Oospora chromogenes. Streptothrix violacea (Oospora violacea, Sauvageau and Radais, and Doria. — This streptothrix liquefies gelatine, and gives it a pale wine-red colour. Agar is coloured a violet tint, and potato becomes a reddish- brown. Urobacillus Dnclauxi (Miquel).— Rods '6 to -8 p in diam., and filaments 2 to 10 /* in length. Motile. Spore-formation present. In gelatine containing ammonia or urea they develop in the track of the needle and cause liquefac- tion. The liquefied gelatine is viscid. Broth containing ammonia be- comes turbid, a sediment forms, and the liquid gives off an unpleasant odour. They occur in sewage. 574 DESCRIPTION OF SPECIES. Urobacillus Freudenreichi (Miquel). — Rods 1 to 1-3 p in width, and filaments 5 to 6 n in length. Colonies circular, white. Inoculated in the depth of gela- tine growth occurs in the track of the needle, and a pure white growth on the surface, followed by slow liquefaction. In broth they produce turbidity. They decompose urea. They occur in air, sewage and dust. Urobacillus Maddoxi (Miquel). —Rods 1 p. in width, 3 to 6 p, in length, and involution forms. Inoculated in the depth of gela- tine containing urea they produce white colonies and crystals. In broth they produce turbidity. They decompose urea. They occur in sewage. Urobacillus Pasteuri (Miquel). — Rods attaining 1-2 /A in width, and 4 to 6 /u. in length, singly and in pairs. Spore-formation present. They grow in ammoniacal gela- tine, slowly liquefying it and form- ing crystals. The liquefied gelatine is viscid. They ferment urine, producing a copious deposit of crystals. They were isolated from decom- posing urine. Urobacillus Schutzenbergi (Miquel). — Short rods '5 p. in width, 1 n in length. They rapidly liquefy gelatine. On agar they form a white layer. They grow readily in broth, especially after the addition of urea. The liquid is made cloudy, but after a few days it becomes clear again. They occur in water. Vibrio rugula (Miiller).— Rods and threads, 6 to 16 p long, about •5 to 2-5 \i thick. The rods are either simply bowed, or possessed of one shallow spiral (Fig. 230). They bear a flagellum at each end. The rods form swarms when caus- ing decomposition, and then, or after, grow out into threads, curved in a screw-like manner. In the FIG. 230.— VIBRIO RUGULA, x 1020. A. Bowed threads. B. Slightly- curved rods. C. Rods swollen pre- paratory to spore-formation. D. Rods swollen at the spore -forming end. E. Various stages of the developing spores. (Prazmowski. ) next stage of development the rods cease to move, and become swollen with granular contents. One ex- tremity develops an enlargement, giving the rod the appearance of a pin. The spore formed by the contraction of the plasma in the swollen end finally becomes globular. The vibrios appear in vegetable infusions, causing fermentation of cellulose. APPENDICES 575 APPENDIX I. YEASTS AND MOULDS. Yeast-fungi and mould-fungi, like bacteria or fission-fungi, are- achlorophyllous Thallophytes. They belong to two separate orders — the Saccharomycetes and Hyphomycetes — which are intimately related to each other, but quite distinct from bacteria. Their germs occur widely distributed in air, soil and water, and are constantly encountered in bacteriological investigations. In addition, many species are of hygienic and pathological interest and importance in being either accidentally associated with, or the cause of various morbid processes and fermentations. For a complete account of all the described species and full details of the various forms of development, reference must be made to botanical and other works.* A description of certain species is appended here, and may afford some useful information to the worker in a bacteriological laboratory. YEAST- FUNGI OR SACCHAROMYCETES. Saccharomyces cerevisise (Torula cerevisice). — Cells round or oval, 8 to 9 //, long, singly or united in small chains. Spores occur three or four together in a mother-cell, 4 to 5 /A in diam. S. cerevisise, S. pastorianus and S. ellipsoideus are active alcoholic ferments. According to Jorgensen they will produce in fourteen days in beer-wort from 4 to 6 per cent., by volume, of alcohol. Saccharomyces ellipsoideus (Hansen). I. — Elliptical cells, mostly 6 /A long, singly or united in little branching chains. Two to four spores found in a mother-cell, 3 to 3*5 //, in diam. Cultivated on the surface of wort-gelatine they produce in eleven to fourteen days, at 25° C., a net-like growth by which they can be recognised * Sachs, Text-book of Botany ; Jorgensen, Micro-organisms and Fermen- tation. 577 37 578 APPENDICES. with the naked eye. II.— Round, oval, and rarely elongated cells. They produce yeast-turbidity. There are two so-called disease-yeasts allied to this species. The colonies of one kind form a network. This yeast causes turbidity in beer, and a bitter after- taste. In the other kind the colonies are sharply denned. It produces a disagreeable aromatic taste to beer, and an astringent after- taste. It is widely distributed, and is the principal agent in .accidental fermentation. Saccharomyces eonglomeratus (Reess). — Cells round, 5 to 6 fji in diam., united in clusters, consisting of numerous cells produced by budding from one or a few mother- cells. There are 2 to 4 spores in each mother-cell. They occur on rotting grapes and in wine at the commencement of fermentation. Saccharomyces exiguus (Reess). — Conical or top-shaped cells, 5 p long, and reaching 2'5 /x, in thickness, in slightly branching colonies. Spore-forming cells are isolated, each contain- ing 2 or 3 spores in a row. They occur in the after-fermentation of beer; but, according to Hansen, they do not produce disease in beer. Saccharomyces Jb'rgensenii (Lasche). — Cells small, round or oval. On the surface of wort-gelatine the culture is greyish- white, and the gelatine is slowly liquefied. They ferment saccharose and dextrose, but not maltose. When grown in wort with other yeasts they are rapidly crowded out. Saccharomyces pastorianus, I. — Cells oval or club-shaped. Colonies consist of primary club-shaped links, 18 to 22 p long, which build lateral, secondary, round or oval daughter-cells, 5 to 6 p long. Spores 2 to 4. They occur in the after- ferment at ion of wine, fruit-wines, or fermenting beer, and in the air of breweries. They produce a bitter taste and unpleasant odour and turbidity in beer. II. — Cells mostly elongated, but also oval or round. Cultivated on the surface of gelatine and yeast- water a growth is produced with smooth edges, by which it can be differentiated from No. III. They occur in the air of breweries, but do not produce disease in beer. III. — They produce yeast-turbidity in beer. On the surface of yeast- water gelatine the cultures, after sixteen days, have hairy edges. Saccharomyces apiculatus. — Cells lemon -shaped, both ends bluntly pointed, 6 to 8 /x long, 2 to 3 p wide. Budding occurs only at the pointed ends. Rarely united in colonies. Spores unknown. They occur with other yeasts in various accidental fermentations -and in ripe fruits. YEASTS AND MOULDS. 579 Saccharomyces sphsericus. — Cells varying in form ; the basal ones of a colony oblong or cylindrical, 10 to 15 /x long, 5 /x thick ; the others, round, 5 to 6 /A in diam. United in ramified families. Spores unknown. Saccharomyces anomalus (Hansen). — Cells small, oval, and sometimes elongated. Spores are hemispherical, with projecting rims at the base. They were found in impure brewery yeast. Saccharomyces mycoderma (Mycoderma, cerevisice et vini). — Cells oval, elliptical, or cylindrical, 6 to 7 /x long, 2 to 3 /x thick, united in richly-branching chains. Spore-forming cells may be 20 /x long. Spores 1 to 4 in each mother-cell. The colonies in gelatine are greyish and filmy. They form the so-called " mould " on fermented liquids, arid develop on the surface without exciting fermentation. When forced to grow submerged, a little alcohol is produced, but the fungus soon dies. They occur on wine, beer, fruit- juices and sauerkraut. Saccharomyces albicans (Oidium albicans, Fungus of thrush). — Cells round, oval, or cylindrical, 3*5 to 5 /x thick ; the cylindrical cells 10 to 20 times as long as they are thick. The bud- colonies mostly consist of rows of cylindrical cells, from the ends of which oval or round cells shoot out. Spores form singly in roundish cells. In plate-cultivations the colonies are pure white. In the depth of gelatine a filament is formed composed of white colonies, some with ray-like processes extending into the gelatine. On potato the fungus forms a rapid white growth, and on bread also. They can be easily cultivated in a nutrient solution containing sugar and ammonic tartrate. The cells germinate according to the rich- ness of the fluid in sugar ; they either grow into long threads, or, in a very strongly saccharine solution, many daughter- cells are formed and bud out in various directions. According to Klemperer the thrush-fungus is pathogenic in rabbits, death taking place twenty-four to forty-eight hours after an intravenous injection of a pure- culture. Long mycelial threads are found in the internal organs. They occur on the mucous membrane of the mouth, especially of infants, in greyish -white patches, which consist of epithelium, bacteria, yeasts, and the mycelia of various moulds. Saccharomyces pyriformis (Marshall Ward). — Cells oval. They convert saccharine solutions containing ginger into ginger- beer. They occur with other micro-organisms in the so-called " ginger-beer plant." Saccharomyces glutinis. — Cells round, oval, or short cylinders, 5 to 11 /x long, 4 /x wide, isolated, or united in twos. 580 APPENDICES. Cell- membrane and contents are colourless in the fresh state, but when dried and re-moistened possess a pale-reddish nucleus in the middle. Spore-formation unknown. They form rose-coloured, slimy spots on starch paste, and on sterilised potatoes. The colouring matter is not changed by acids or alkalies. Saccharomyces ilieis (Gronlund).— Cells spherical. Spore- formation present without vacuoles. Cultures on the surface of gelatine have a powdery appearance. They produce about 2 '8 per- cent.,. by volume, of alcohol in beer-wort, and cause a disagree- able, bitter taste. They were obtained from the fruit of Ilex aquifolium. Saccharomyces aquifolii (Gronlund). — Cells large and spherical. Spores contain vacuoles. Cultures on gelatine are variable, smooth and shining, or powdery. They produce about 3-7 per cent, alcohol in beer- wort, and cause a sweet taste with bitter after-taste. They also were obtained from the fruit of Ilex aquifolium. Saccharomyces Marxianus (Hansen). — Cells elongated. They develop a mycelial growth on solid nutrient media. They occur on grapes. Saccharomyces membransefaeiens (Hansen). — Cells elon- gated and vacuolated. Spore-formation abundant. Cultivated on wort-gelatine they produce circular, flattened and wrinkled colonies, greyish, and sometimes with a reddish tinge. The gelatine is slowly liquefied. They occur in the slimy secretion of the roots of the elm, and were also isolated from well-water. Saccharomyces Hansenii (Zopf). — Cells with small spherical spores. They set up alcoholic fermentation in solutions containing sugar. They were found in cotton- seed flour. Saccharomyces Ludwigii. — Cells irregular in form, oval, bottle-shaped, lemon-shaped, and elongated, and mycelial filaments. On wort- gelatine the growth is greyish or yellowish. Saccharomyces acidi lactici (Grotenfelt). — Cells oval, 2 to 4-35 /A in length, and 1'5 to 2'9 /x in width. Colonies on nutrient gelatine are porcelain-white. They coagulate milk. Saccharomyces minor (Engel). — Cells spherical. Spore- formation present. They are said to be the most active ferment in the fermentation of bread. Saccharomyces rosaceus (Pink Torula). — Cells 9 to 10 /x in diam. They form a coral- pink growth in nutrient gelatine, nutrient agar-agar, or on sterilised potatoes. They are present in the air. YEASTS AND MOULDS. 581 Saccharomyces niger (Black Torula).— Cells also present in the air. Cultivated in nutrient gelatine they form a black crust (Fig. 231). MOULD-FUNGI OR HYPHOMYCETES. The mould-fungi have been divided into five orders : Hypodermii, Phycomycetes, Asco- mycetes, Basidiomycetes and Myxomycetes. The following species, with the orders to which they belong, are of especial interest : — HYPODERMII. Ustilago carbo (Mildew, Smut). — Spores brown, circular ; episporium smooth ; sporidia, ovoid cells. The spores or conidia occur as a black powder in the ears and panicles of wheat, barley and oats. Tilletia caries. — Spores round, pale brown ; episporium with reticulated thicken- ings. In germinating, the sporidia grow out radially from the end of the promyce- lium ; these, at their lower part, conjugate p^ 231 _BLACR TQR. by a cross branch and separate from the ULA> pURE CULTIVA- promycelium, and at some point of the pair TION ON POTATO. a hypha grows out, on which abundant secondary sporidia develop. The latter are long, oval cells, which can in turn germinate. The fungus occurs in the form of a stinking powder in grains of wheat, which renders the meal im- pure, and gives it a disagreeable smell. Urocystis occulta. — The spores consist of several cells united together; partly, large dark-brown cells in the interior, and out- side, several flat, semicircular, colourless cells. The promycelium germinates as in Tilletia, but the cylindrical cells produce a hypha, without, as a rule, previous conjugation. They occur as a black powder in rye-straw in long disintegrated stripes, which are at first greyish. The affected plant produces abortive ears. Empusa muscse. — A spore or conidium of this fungus alighting upon the white area of the under surface of the body of the house-fly germinates into a hypha. The latter, penetrating the skin, forms toruloid cells, which multiply by germination, and are disseminated in the blood throughout the body of the fly. 582 APPENDICES. These cells again grow into hyphse, which penetrate the skin, each forming a conidium, which is cast off with considerable force. The parasite is fatal to flies, especially in the autumn. They are often observed attached to the walls or window-panes, surrounded by a powdery substance, consisting of the extruded conidia. Empusa radicans. — The spores form long hyphse, which pierce the transparent skin of the caterpillar of the cabbage white butter- fly. The terminal cells ramify, and fill the body of the caterpillar with a network of mycelial filaments. The caterpillars attacked become restless, then motionless, and death ensues. Tarichium megaspernmm. — The spores are black in colour, and provided with a thickened episporium. They occur at the sides and ends of mycelial threads, attacking caterpillars (Agrotis segetum). PHYCOMYCETES. Saprolegnia. — Colourless threads, forming dense radiating tufts, occur on living and dead animal and vegetable matter in fresh water. The filaments penetrate into the substratum, and branch more or less in the surrounding water. The cylindrical ends of the threads are shut off by a septum — forming zoosporangia, or mother- cells, in the interior of which a number of spherical zoospores develop. These are set free through an apical opening in the thread, and after a time coming to rest, give rise to new plants. In the sexual mode of reproduction a spherical bud, the oogonium, develops at the end of a mycelial thread; from the thread small processes or antheridia sprout out laterally towards the oogonium and blend with its protoplasm. The latter breaks up into a number of oospores, which clothe themselves with a membrane while still within the mother- cell, and, eventually being set free, grow into fresh mycelial filaments. The fungus attacks fish and tritons, and produces a diseased condition of the skin, which may be ultimately fatal. In salmon it produces the common "disease of salmon." Peronospora infestans. — The conidia -bearers of this fungus have as many as five branches, each bearing an egg-shaped conidium. The contents of the conidia falling off and reaching a drop of moisture, break up into a number of swarming zoogonidia, which in turn develop upon plants. Fixing themselves to the cuticle of the host, they throw a germinating filament into an epidermal cell ; after piercing first its outer wall, and then its inner YEASTS AND MOULDS. 583- wall, the filament reaches an intercellular space, where the mycelium develops. This continues to grow and spread throughout the plant. In tubers it can hibernate and develop in the young shoots in the following spring. The fungus appears in the form of brown patches on the green parts of the plants, especially the leaves. The attacked parts wither and turn yellow or brown in colour. If the under surface of a diseased leaf is examined, a corresponding dark spot may be observed, accompanied with a faint greyish-white bloom, which covers it. The latter consists of the conidia- bearing branches. Pilobolus. — The fruit -hyphse possess spherical receptacles containing conidia. When ripe the receptacles with their conidia are detached at their bases, and spring by their elasticity to some distance. The fungus occurs as glassy tufts on the excrement of cows, horses, etc. A cultivation can generally be obtained by keeping fresh horse-dung under a bell-glass. Mucor mucedo. — Hyphae colourless, simple or branched ; spo- rangia yellowish-brown or black ; spores ovoid. They form the familiar white mould on fruits, bread, potatoes and excreta, and penetrate into the interior of nuts and apples. A network of fibrils develops in the substance of nutrient gelatine, with forma- tion of sporangia on the free surface. The germination of the spores and development into hyphse can be observed in a few hours if the fungus be cultivated in a decoction of horse-dung. Mucor racemosus. — Hyphse short ; sporangia, yellowish to pale -brown ; spores round. By continued cultivation in liquids saturated with carbonic acid, the hyphse become still shorter arid exhibit a yeast-like sprouting. These yeast-like or toruloid cells can, when the carbonic acid is withdrawn, germinate into normal mycelium. They occur on bread and decaying vegetable matter. Mucor stolonifer (Lichtheiin). — Mycelium grows in the air and then bends down and re-enters the nutrient substratum ; sporangia black, and spores globular. The mycelium can penetrate through the shell of eggs, and form conidiophores within them. Mucor aspergillus (Lichtheim). — Fruit-hyphae thinned at the base, and with many fork-like divisions ; dark-brown spores. Mucor phycomyces (Lichtheim). — Mycelium thick-walled ; olive-green fruit-hyphse ; black sporangia, and oblong spores. Mucor macroearpus (Lichtheim). — Spindle-formed, pointed spores. Mucor fusiger (Lichtheim). — Ovoid spores. 584 APPENDICES. Mucor mellittophorus (Lichtheim). — Spores elliptical. Found in the stomach of bees. Mucor corymbifer (Lichtheim). — This fungus forms branched fruit-hyphae. The sporangia have a smooth membrane. It has been found in the external auditory meatus, and on bread it forms a dense snow-white growth. Pathogenic in rabbits. Mucor rhizopodiformis (Lichtheim). — The spores of Mucor rhizopodiformis and Mucor corymbifer, when introduced into the vascular system of rabbits, can germinate in the tissues, especially in .the kidneys, where they set up haemorrhagic inflammation. Dogs are immune, and only artificial mycosis is known. It occurs on bread. Mucor erectus. — Resembles Mucor racemosus. It occurs on rotting potatoes. Mucor circinelloides. — Mycelium much branched, and sporangium carrier is curved. Mucor spinosus. — Sporangia chocolate. Columella has short processes or spines. ASCOMYCETES. Oidium Tuckeri. — Fruit-hyphae bearing single ovoid conidia. Observed in the form of brown patches, covered with a white mildew- like layer on the leaves, branches and young fruit of the vine, producing " grape disease." Oidium lactis. — Fruit-hyphae simple, erect and colourless, bearing at their ends a series or chain of conidia. In some cases, the fruit-hypha branches beneath the chain of spores. Spores are short cylinders. The conidia germinate into filaments of varying length, which by subdivision form septate mycelial hyphse ; these and their branches give rise in turn to spores or conidia. The fungus is deeply stained by the ordinary aniline dyes. In a plate- cultivation the colonies appear as white points, and develop into delicate stellate colonies which ultimately coalesce and form a fine mycelial network covering the surface of the gelatine. The gela- tine is not liquefied. The growth on the surface of agar is similar to that on gelatine. The fungus occurs in sour milk. Achorion Schonleinii (Fungus of favus).— Threads branching at right angles. Favus in man forms yellow crusts on the hairy parts of the body. The crusts are composed of epidermis and mycelial filaments and spores. In plate-cultivations whitish colonies are formed surrounded by liquefied gelatine. Cultivated on the surface YEASTS AND MOULDS. 585 of gelatine the growth resembles that of Tricophyton tonsurans, but the liquefaction takes place more slowly, and there is a more distinct yellow colour. On agar the growth is white, dry and firmly adherent. Tricophyton tonsurans (Fungus of ringworm).— Mycelial filaments and spores occur on the crusts and in diseased hairs. In plate-cultivations white colonies are formed, and liquefaction quickly follows. In test-tube cultivations the gelatine is liquefied FIG. 232.— HEAD AND NECK OF CALF WITH ADVANCED RINGWORM (Broiori). and the fungus forms a membrane on the liquid jelly which is white above and yellow beneath. The surface of the growth is powdery In man the disease varies in appearance in different parts of the body. Cattle, horses and dogs also suffer from ringworm ; but sheep and pigs rarely, if ever. The disease is very common in calves. Sometimes a small portion of the skin is diseased; in other cases, the head, neck, chest and abdomen, or even the whole trunk, may be covered with scabs or crusts. There is often loss of hair in patches, and the skin may be covered with scurf. The disease is transmissible 586 APPENDICES. to the human subject. In one case, according to Brown, seven grooms were infected on the arms from a grey pony which was suffering from the disease in an aggravated form. Fungus of fowl-scab. — Fowls are liable to a disease similar to favus. According to Schiitz this disease is characterised by greyish-white patches on the comb and wattles of fowls, which may extend over the neck and body. On nutrient gelatine a white mycelium is formed ; and the gelatine is liquefied, and acquires a reddish tint. The fungus can be readily cultivated on bread-paste, agar-agar and potato. Cultures inoculated in fowls produce the disease, but have no effect on mice and rabbits. Fungus of mouse-favus. — Mice suffer from a form of favus which can be communicated to healthy mice by inoculation of scabs or infected skin (Nicolaier). On nutrient agar the fungus forms a thick mycelium, at first white, and later of a red or reddish-brown colour. Mice can be infected with cultures. Microsporon furfur. — This fungus occurs in Pityriasis versicolor. Grawitz regarded it as identical with Oidium lactis, and it is very closely related. Cultivated on gelatine the jelly is hollowed out and the mycelial growth sinks down, and is yellowish in colour. Oidium. albicans. — Vide Saccharomyces albicans. Aspergillus glaucus (Eurotium aspergillus glaucus). — Mycelium at first whitish, becoming grey-green or yellow-green. Spores grey-green, thick -walled. It is found on various substances, chiefly cooked fruit, and is non- pathogenic. Aspergillus repens (Eurotium repens, De Bary). — Fruit-heads fewer than in the above, which are at first pale and then blue-green to dark-green in colour. Conidia mostly oval, smooth, colourless or pale to grey-green. Aspergillus flavus.— Gold-yellow, greenish and brown tufts. Fruit-heads round, yellow, olive-green or brown. Conidia round, seldom oval, sulphur-yellow to brown in colour. Saprophytic in man, pathogenic in rabbits. Aspergillus fumigatus. — Greenish, bluish or grey tufts. Fruit-heads long and conical. Conidia round, and rarely oval, smooth, mostly pale and colourless, This fungus occurs on bread,, and has been found in the human lungs, external auditory meatus and middle ear, and in the lungs of birds. The spores introduced into the vascular system of rabbits, or into the peritoneal cavity, establish metastatic foci in the kidneys, liver, intestines,, lungs, muscles, and sometimes in the spleen, bones, lymphatic glands, nervous system and skin. YEASTS AND MOULDS. 587 Aspergillus niger (Eurotium aspergillus niger, De Bary).— Dark chocolate-brown tufts. Conidia round, black-brown, or grey- brown when ripe. This mould can be cultivated readily on bread moistened with vinegar, on slices of lemon, and on acid fruits and liquids. It nourishes best of all, according to Raulin, in a liquid of the following composition : — Grammes. Water 1500' Sugar-candy ....... 70* Tartaric acid ....... 4* Nitrate of ammonia ...... 4' Phosphate -6 Carbonate of potassium ..... '6 „ ,, magnesium ..... '4 Sulphate of ammonia . . . . . '25 ,, ,, zinc ...... '07 „ iron -07 Silicate of potassium ..... '07 It was also found that the fungus grew best when the liquid was spread out in a layer 2 or 3 cm. in depth in a shallow dish ; and a temperature of 35° C. proved to be the most favourable. The abstraction of zinc from the, nutritive liquid reduced the weight of a crop from 25 (the average) to 2 grammes, and the presence of T^OOOOO Part of nitrate of silver, or ^-^ part of corrosive sublimate, stopped the growth altogether. It is saprophytic in the living body. METHOD OF EXAMINING ASPERGILLUS NIGER. Species of aspergillus stain intensely with carmine, f uchsine or methyl- violet ; but to examine Aspergillus niger with a high power a little special technique is employed, as follows : — A drop of glycerine is placed on a clean slide, and a drop of alcohol on a cover-glass. With a fine pair of forceps a few of the fruit-hyphae with their black heads are immersed in the alcohol. The cover-glass is then turned over on to the drop of glycerine, and the slide held in the flame of a Bunsen burner till the spores or conidia are dispersed. To make a permanent preparation remove the cover-glass, and transfer the fruit-hyphae so treated to a mixture of glycerine and water (1 to 5) ; a drop may be conveniently placed ready on a slide provided with a ring of Canada balsam. The specimen is then, permanently mounted by employing a circular cover- glass, and surrounding it with a ring of cement in the usual way. 588 APPENDICES. Aspergillus ochraceus. — At first flesh-coloured, and then ochre-yellow heads. Aspergillus albus. — Pure-white fruit-heads. Aspergillus clavatus. — Club-shaped fruit-heads on long stems. Aspergillus nidulans. — Bread and potatoes acquire a reddish- brown colour. Pathogenic in rabbits. Occurs on bread. Aspergillus subfuscus. — The growth is olive-yellow in colour. Pathogenic in rabbits. Occurs on bread. Aspergillus flavescens. — The growth is yellowish-green. Pathogenic in dogs and rabbits. Occurs on bread. Penicillium glaucum. — Occurs as a white, and later a blue green, mould, on which dew-like drops of liquid may appear. Its spores are present in large numbers in the air, and are liable to contaminate cultivations. The fruit-hypha bears terminally a number of branched cylindrical cells, from which chains of greenish conidia are developed. It is the commonest of all moulds. Botrytis Bassiana — Hyphse and spores colourless. Hyphae usually simple, but sometimes united in arborescent stems. It is the cause of muscardine, a fatal disease of silkworms, and occurs also in various other caterpillars and insects. Chionyphe Carteri.— Mycelial filaments observed by Carter in Madura disease. APPENDIX II. H^EMATOZOA. H^MATOZOA IN MAN, BIRDS AND TURTLES. — H^MATOZOA IN EQUINES, CAMELS, RATS AND FISH. — ILEMATOZOA IN FROGS. H^EMATOZOA IN MAN (MALARIA). IN 1880 Laveran, in Algiers, noticed the existence of peculiar structures in the blood of a patient suffering from malaria, and his researches were communicated to the Academy of Medicine in Paris in 1881 and 1882, and subsequently published in extenso in a treatise on the subject. Laveran described various bodies which he was led to' regard as different stages in the life-history of the same micro-parasite. The most striking forms were cylindrical elements with pointed extre- mities. They were crescent -shaped and pigmented in the middle. There were other forms, more frequently found, which were either free in the serum or in contact with the red blood-corpuscles. They were more or less spherical, pigm exited, and endowed with amoeboid movement. Other forms, again, were provided with motile filaments three or four times as long as the diameter of a red blood- corpuscle. And, lastly, there were little masses of hyaline material, which Laveran regarded as dead forms. These observations at first attracted little attention ; but they have since been confirmed and extended by Richard, Councilman and Abbot, Marchiafava and Celli, Golgi, Sternberg, Osier, the author, Vandyke Carter, Manson, and others, and their importance fully recognised. The different forms assumed by the haematozoon of malaria may be described in two groups : those within the red blood-corpuscles, and those free in the serum. Intra-corpuscular bodies. — These are of three kinds. First t 589 590 APPENDICES. structureless protoplasmic bodies much smaller than, and within or attached to, the red blood- corpuscles (Fig. 233). These rapidly change their shape, exhibiting amoeboid movement. They were first described by Marchiafava and Celli, and possibly represent the first stage in the life -history of the hsematozoon. Marchiafava and Celli suggested the name Plasmodium malarise. Second, minute FIG. 233.— NON-PIGMENTED AMEBOID FoEMS (Marchiafava and Celli). masses of finely granular or of hyaline protoplasm enclosing granules of pigment (Fig. 234). These forms are sometimes present in large numbers, and at other times can be found only with difficulty. They are more or less spherical, but exhibit amoeboid movement, and rapidly change their form. The pigment granules are also in active movement. There may be one or more of these amoeboid FlG. 234.— PlGMENTED AM(EBOID FORMS (Golgi). bodies to a blood-corpuscle, and they vary in size ; one may occupy the whole of the corpuscle. In cases of pernicious malaria, similar bodies may be seen, in tissue sections, in the corpuscles filling the capillaries. Third, forms which appear like isolated grains, and larger homogeneous bodies surrounded by clear spaces which change in outline. Extra-corpuscular bodies.— These are the most striking, and FIG. 235.— SEMI-LUNAR BODIES OF LAVERAN (Golgi). perhaps the most interesting, forms. First, the semi-lunar bodies Laveran. These are crescent -shaped bodies, sometimes pointed ANIMAL MICRO-PARASITES. 591 at the extremities, but more usually rounded off (Fig. 235). They are not always curved ; some, indeed, are almost spherical, and others sausage-shaped. They are motionless. In many specimens a delicate line is visible on the concave side of the crescent connect- ing the extremities. On careful examination this is found to be 01 .€> O FIG. 236.— ROSETTE FORMS WITH SEGMENTATION (Golgi). the edge of a very delicate membrane. The body is composed of homogeneous protoplasm. Centrally placed is a collection of pigment granules, which on careful examination can be distinctly seen to be in movement. The semi-lunar bodies vary in number in different cases. Sometimes several can be seen in the field at the same time, and in other cases they are only observed after a long and patient search. They are, as a rule, free in the serum ; but they have also been seen within the red blood-cells. Second, finely granular masses of protoplasm, which arise, according to Golgi, from the intra- oorpuscular pigmented bodies. The pigment is collected in a rosette, and the protoplasm by segmentation gives rise to a number of small FIG. 237. — FLAGELLATED FORMS (Vandyke Carter). 1. A flagellated spherule ; a, the same in the interior of a phagocyte ; 6, free motile filaments. spherical forms, which are ultimately set free (Fig. 236). Golgi believes that these changes occur in definite relation to the develop- ment of the paroxysm. Third, spherical, pear-shaped, or ovoid bodies, rather smaller than the red blood-corpuscles, and provided with one or more actively motile flagella (Fig. 237). These flagella 592 APPENDICES. are long lash-like filaments, which by their activity set the neigh- bouring blood- corpuscles in motion. Free filaments in active move- ment have also been observed. Fourth, small spherical pigmeiited bodies about one-quarter the size of a red blood-corpuscle, which exhibit amoeboid movement. Inoculation experiments. — Marchiafava and Celli assert that inoculation of a healthy subject with blood containing the parasites will produce a paroxysm of ague with development of the haematozoa. The pathogenic power of these parasites, however, has not been established. There has been no cultivation of the parasite outside the animal body, and reproduction of the disease with a pure culti- vation. In favour of its being a pathogenic organism, Laveran points out its invariable presence in some form or other in cases of malaria ; the marked changes it effects in the red blood-cells ; the increase in the number of the parasites in proportion to the severity of the attack ; and, lastly, their disappearance after the admini- stration of quinine. Others, again, have doubted the parasitic nature of these bodies, and have looked upon them as representing pathological changes in the blood- cells. Laveran first of all suggested the name Oscillaria malarias ; but subsequently he recognised that these bodies belonged to the animal, not to the vegetable, kingdom. Osier has suggested that, tempo- rarily at any rate, the organism should be placed in the genus Hsematomonas of Mitrophanow, thus : " Genus, Hsematomonas ; species, Hsematomonas malaria. Definition — Body plastic ; ovoid or globose ; no differentiation of protoplasm, which contains pigment grains ; flagella variable, from one to four ; highly polymorphic, occurring in (1) amoeboid form, (2) crescents, encysted form, (3) sporocysts, (4) cellular free pigmented bodies." EXAMINATION OF THE HAEMATOZOA OF LAVERAN. In the Living Condition. — Select a patient by preference who has had several attacks of malaria, and is markedly ansemic. Examine before the invasion of the febrile paroxysm. Take two perfectly clean cover-glasses and two clean slides ; wash one of the fingers of the patient with soap and water, and then cleanse with alcohol ; apply a ligature, and with a clean needle puncture the thin skin near the root of the nail ; touch the drop of blood which collects, with a clean slide ; cover quickly with a cover-glass, and gently press it if the layer of blood be too thick. Examine with a & o. i. In Stained Preparations.— Puncture the finger again if necessary ; touch the droplet of blood with a clean cover-glass ; apply another cover- glass ; press them gently together, and then slide them apart ; stain with ANIMAL MICRO-PARASITES. 593- two or three drops of alcoholic solution of methylene-blue ; wash off excess, and examine in water, or allow the preparation to dry, and mount in balsam. H^EMATOZOA OF BlRDS. According to Danilewsky, birds suffer from malaria in both an acute and a chronic form. The haematozoa are very similar to those found in malaria in man, and any slight difference may be attributed to the different character of the blood in birds. Grassi and Feletti have described two kinds of malarial hsematozoa in birds, one kind belonging to the genus Hcemamveba and the other to the genus Laverania. H^MATOZOA OF TURTLES. Danilewsky has also minutely described and figured hsematozoa in the blood of turtles, which in some stages of their life history very closely resemble those found by Laveran in man. H^MATOZOA OF EQUINES AND CAMELS (SURRA DISEASE.) Swra is a blood disease occurring in horses, mules, and camels,, characterised by fever accompanied by jaundice, petechise of mucous membranes, great prostration, and rapid wasting terminating in death. The average duration of the disease is about two months. No organic lesions are found after death, but a parasite exists in the blood during life. By means of subcutaneous inoculation, and by the introduction into the stomach of blood containing the parasite, the disease, according to Evans, can be transmitted to healthy animals. The importance of this disease may be realised from the fact that on one occasion in India the 3rd Punjab Cavalry lost no less than three hundred horses from it. The disease has not been observed to be contagious or infectious in the ordinary sense, but the possibility of its conveyance by means of large brown flies has been suggested. These flies attack the horses so vehemently that the blood frequently streams from the bites ; and the opinion that they propagate the disease is prevalent among the natives. At the same time it has been particularly noted that where the disease has broken out the water was very impure. Evans discovered a haematozoon, in 1880, in all the diseased horses and mules examined ; in all diseased camels, with one exception ; and in the dogs which had been subjected to experimental inoculations 38 594 APPENDICES. Evans stated that when he first discovered the parasite he thought it was a spirillum, but very speedily on closer examination arrived at an opposite opinion. To him the organism presented the appearance, when fresh and active, of an apparently round body, tapering in front to form a neck and terminating in a blunt head. Posteriorly he described a tapering tail, from which there extended a long slender lash. At the head end there appeared in one or two cases a circlet of pseudopods, and as the body slowly died in serum it gave the appearance of flattening out. After watching very closely all its •changes of form and movements, Evans came to the conclusion that there existed on either side of the body two fin-like papillae, one near where the neck began an(J the other close to where the tail began. In only very few instances he was able to see the four at once. He suggested that these processes were of the nature of pseudopods. The parasite he described as extremely active in its movements, with an undulatory, eel-like motion, progressing for the most part head-end foremost, but occasionally moving in the direction of the lash when tugging at a corpuscle. In fresh blood these organisms resembled spermatozoa in colour ; but their peculiar characteristic was the power they possessed of attacking and disintegrating the red corpuscles. Occasionally two were observed to unite and swim off as one body ; but the mode of union was a disputed point. Evans thought that they joined with their respective heads and tails in the same direction, overlapping each other; but others to whom they were shown were of opinion that they fastened with their tails in opposite directions. The parasites were not always present in the blood, but were •observed to come and go in successive broods. Evans referred the organism to Lewis for his opinion as to its nature. Lewis arrived at the conclusion that the parasite was " more nearly related to that which he found in the blood of rats than to any other " ; but he was of opinion at the time that they did not appear exactly the same. Five years later Surra broke out in British Burma, and Steel was deputed to investigate the outbreak. Steel confirmed the communicability of the disease to dogs, horses and mules by ingestion and inoculation, but he considerably supplemented Evans' views as to the nature of the disease by careful thermometric observations : these finally led him to regard the disease as a true ANIMAL MICRO-PARASITES. 595 relapsing fever, closely resembling relapsing fever in man. At the same time it is worth recording that until Steel observed the presence of the parasite described by Evans he regarded the out- break as malarious in origin, and provisionally termed it gastric typhoid. In the Burma outbreak, as in the Punjab epidemic, considerable evidence was adduced in favour of regarding the disease as being due to bad water supply. Steel succeeded in staining the organism with aniline dyes, but his description of the parasite in the frssh state differs very materially from that given by Evans. Steel failed to recognise the round body tapering in front to a neck. To him the bodies appeared thick in the middle, gradually diminishing in size in either direction, with a blunt and rigid extremity at one end. The opposite end he described as tapering in such a way as to produce a subspiral prolongation, which was uncurled and lashed about freely like a whip. This tail was described as slender in relation to the general size of the parasite ; but under the highest power available the presence of a colourless nagellum could not be detected, nor, he adds, did the movements of the blood -constituents indicate its existence. Steel also failed to see the slightest sign of the two fin -like papillae on each side as described by Evans — an opinion in which he was supported by Lewis. These two observers, Evans and Steel, also differed as to whether the movement could be called spiral. Steel felt convinced that their movement was as much of that nature at times as can be expected from organisms with so open a corkscrew shape ; while Evans maintained an opposite view. In the dried and stained specimens Steel observed that they retained their subspiral form of body and markedly spiral form of tail. Steel found that the disease could be communicated to the dog and to the monkey, and then discussed the resemblance of the parasite to the spirillum of relapsing fever in man. From the different appearances presented by the parasite when in the living state and when dried and stained, Steel thought that there was probably a still closer resemblance to the living spirillum than to the dried and stained one, and argued that the figures of spirilla like corkscrews must be purely imaginary. Steel, it must be observed, founded these remarks upon figures in text- books, and not on photographs or on a practical acquaintance with the spirillum of relapsing fever. One cannot refrain from pointing out the value of photomicrographs, for they cannot be 596 APPENDICES. called into question; and had Steel studied photographs of spirilla he would not have regarded the corkscrew appearance as imaghiM ry. Steel found the parasite in all cases, and further observed that it appeared as the temperature rose and disappeared during the apyrexial periods. From all these observations Steel concluded as follows : — That relapsing fever of mules is an invariably fatal disorder, characterised by the periodical occurrence of attacks of high fever, during which a special organism closely resembling the spirillum of relapsing fever in man is found in the blood. This organism is one-sixth the size of a red corpuscle in width and three to six times in length. It is eel-like, and, when dried and stained, presents a thick portion — the body — and a spiral tail. The latter takes less of the dye than the former, and commences as a sudden narrowing of the body, termi- nating by a fine point. This, he insisted, had nothing of the nature of an infusorian nagellum. The thick portion tapers in either direction from its centre, and terminates in front abruptly in a rigid process, with probably some holdfast organ. The sharpness of the head end varies in different animals. The body portion he described as spiral, and so closely in general appearances to resemble the spirillum of relapsing fever that he concluded that the organism was undoubtedly a spiral bacterium and named it after its discoverer Spirochceta Evansi. This view, however, would not be accepted by Evans, who maintained that, whatever it might be, it was not a member of the family of bacteria. In the face of these conflicting opinions Evans, in 1885, submitted to the author preparations of the organism in the blood as well as material from the lungs and intestines of a camel that had succumbed to the disease. On examining a stained preparation the author found that with a power of 200 diameters a number of the parasites could be distinguished in the field of the microscope, and with TV and Y\r o. i. objectives the individual characteristics were clearly brought out. These were quite sufficient at once to dispel the idea of its being a spirillum. It was obvious that it was a more highly organized micro-parasite, presenting very peculiar and distinctive structural appearances. The author came to the following conclusions : — The somewhat tapering central portion, or body, of the parasite is continuous at one end with a whip-like lash, and at the other end terminates in an acutely-pointed stiff filament or spine-like process. ANIMAL MICRO-PARASITES. 597 Here and there, possibly from injury or want of development, the spine-like process appears to be blunted or absent. By very careful focussing on the upper edge of the central portion, the author discovered the existence — much more markedly in some of the parasites than in others — of a longitudinal membrane with either a straight or undulating margin. The membrane is attached along the body, arising from the base of the rigid filament, and becomes directly continuous at the opposite end with the flagellum. In some cases the edge only is deeply stained, giving the appearance of a thread continuous with the flagellum, so that one might be easily led to overlook the membrane, and imagine that the flagellum arose from the opposite end of the body, at the base of the spine-like process. Close to the base of the spine-like process a clear unstained spot is in many parasites easily distinguished ; and at the opposite end there is, in some, the appearance of the deeply-stained protoplasmic contents having contracted within the faintly-stained membranous investment. When the longitudinal membrane has a wavy outline the undulations are much more marked in some cases than in others. Here and there the wavy outline appears first on the one side of the central portion and then on the other ; but there never is any waving outline on both sides of the same part of the body, and this was explained by a careful examination, which showed that the somewhat ribbon -like parasite had become doubled on itself. The discovery of this undulating membrane at once suggested to the author an explanation of the lateral pseudopodia described by Evans. If we imagine that we are looking down upon the parasite, with the edge of the membrane towards us, one can conceive that the rapid undulations, first on one side and then on another, might give an image upon the retina which could be construed as due to the protrusion of lateral pseudopodia. In stained preparations no trace of the jcirclet of pseudopods could be discovered, and the undulating membrane may account for this appearance also. Owing to the somewhat curved and twisted shape of the parasite and the curling of the flagellum in the stained preparations, it was difficult to make exact measurements ; but the average width, according to whether the membrane was visible or not, varied from 1 to 2 /x, and the length of the body from 20 to 30 /M. The flagellum was about the same length as the body. Here and there in a stained preparation there were the forms already described by Evans resulting from the fusion of two para- 598 APPENDICES. sites. But the union obviously took place by the noil- flagellated ends, for the two flagella were frequently turned in the same direction, so that the fused parasites resembled, as Evans sub- sequently suggested, a trophy of buffalo horns. Here and there more than two parasites had united, forming a stellate group ; and in one case the author noticed that the individuals had apparently united with their non-flagellated ends just overlapping, so that the unstained spot in one was just situated in a line with the unstained spot of the other. In Evans's Report, Lewis's opinion is given that these parasites differed slightly, but still were closely allied to certain flagellated organisms which had been observed by him in rats in India. On FIG. 238.— "SURRA" PARASITES OCCURRING SINGLY AND FUSED. (From preparations stained with magenta, x 1200. Lent by Dr. Evans.) referring to his original memoir, it will be found that his description and woodcut differed very materially from the Surra parasite as just described, though a microphotograph which Lewis had appended to the memoir after it was written, indicated a great similarity to this organism. To the author the organisms appeared not only closely allied, but, as far as one can judge from figures and descrip- tions, morphologically identical with the parasites described by Mitrophanow in the carp, and as a matter of fact, instead of a mere resemblance, the rat and the Surra parasites, when stained, are found to be morphologically identical. •-, ANIMAL MICRO-PARASITES. 599 H^MATOZOA OF RATS. In describing these organisms, Lewis remarked that it wa& strange that they had not occupied attention before, and suggested as an explanation that possibly European rats did not harbour these parasites. The author examined a few white rats, but without success, and then proceeded to examine the blood of common brown rats, trapped from the London sewers, and discovered that these organisms are to be found in no less than 25 per cent, of apparently healthy animals. The first question which naturally arose was whether these organisms in European rats were identical with those described by Lewis in Indian rats. If we refer to the description given by Lewis, we find that he states that when he first noticed them he thought they were vibrios or spirilla, The drop of blood under examination appeared to quiver with life ; and on diluting the blood, motile filaments could be seen rushing through the serum and tossing the blood- corpuscles about in all directions. The filaments were pale and translucent, without any trace of visible structure or granu- larity, and they were more un- dulatory in movement than FlG. 239.._pARASITE8 IN THK BLOOD spirilla. A corpuscle might be OP RATS (Lewis), observed to quiver, and this could be distinctly traced to be due to the existence of a flagellum, apparently a posterior flagellum, as the organisms seemed generally to move with the thicker end forward; no flagellum could be detected at the opposite end. The greater number of the figures- in the woodcut (Fig. 239) are described as representing these organisms a few hours after the blood had been obtained, when their movements are not so rapid, and the flagellum becomes more easily recognisable. This observation led Kent, who named the organism Herpetomonas Lewisi, to remark that if, as Lewis is inclined to maintain, that organ " propels instead of draws the animalcule through the in- habited serum, we have presented a structural and functional feature without parallel among the other representatives of these Protozoa flagellata, the recognition of which would demand the creation of a APPENDICES. distinct generic and family group for the reception of these singular organisms." In his later paper, however, Lewis came to the con- elusion that, like the generality of flagellated organisms, the rat parasites moved with the lash in front. On careful examination the plasma which constituted the thicker portion of their substance was observed to suddenly swell out so as to divide the body into two parts, as seen in the centre of the figure ; .at other times two or three such constrictions or dilatations were detected, and at other times the body assumed an arrow shape, as depicted at the lower part of the figure. When dried, and stained with a little weak solution of aniline-blue, the body presented a very different appearance. It was found to have contracted irregularly, and to manifest a somewhat granular and shreddy appearance, .suggestive of a coagulated fibro-albuminous substance. The body portion became flattened towards its middle to double its original width, and both ends almost acutely pointed, while the flagellum was only partly visible. After fixing with osmic acid they measured •0*8 to 1 ju, in width, 'and 20 to 30 ft in length ; the flagellum was about as long as the body : so that the total length of the organism was .about 50 fj.. Lewis detected these parasites in 29 per cent, of the .species Mus decumanus and Mus rufescens, but failed to find them in mice. He considered that they had many features in common with motile organisms of vegetable origin ; but they appeared to approach much more closely to the Protozoa, more particularly several of the species of Dujardin's Cercomonas. He points out that many, however, believe that these organisms are zoospores and not animalcules. To him they also seemed to be not unlike the flagellated parasite described by Biitschli. The latter observer detected flagellated organisms (Leptomonas liutschlii) in the intestinal canal of a free nematode (Trilobus gracilis). They, too, form stellate colonies, like the Surra parasite, owing to their being attached by their non- flagellated ends. When detached from these colonies they presented a somewhat spindle- shaped body about 11 ft in length, with a somewhat thick flagellum about double this length, so that the total length of the protozoon would be 33 /w,, or, as Lewis states, about half the length of the flagellated organism in the rat's blood. Near the base of the flagellum, Butschli's protozoon presented a contractile vacuole, but Lewis was unable to detect any such vacuole in the rat haematozoa. In conclusion, Lewis observed that very probably these organ- isms corresponded with the vermicides observed by Goss in the ANIMAL MICRO-PARASITES. 601 blood of a field mouse, and he also mentions that Chaussat found minute " nematodes " in the blood of a black rat. Wittich discovered in the blood of hamsters whip- like bodies with lively movements. They resembled frog's spermatozoa, possessing a thick portion continued into a long lash -like thread. Wittich considered them identical with the organisms described by Lewis, and they also were observed in apparently healthy animals. Koch later met with the same organisms. Like Lewis, the author found that the blood of the common brown rat in England appeared to quiver with life, and that the parasites were extremely difficult to examine until their movement was arrested for a moment or they became imprisoned in the serum areas. After examining with various powers, from a -I dry to a ^v o. i. of Powell and Lealand, the author came to the following conclusion : — That they are polymorphic, presenting for FIG. 240.— A MONAD IN RAT'S BLOOD. The organism is represented at partial rest with its posterior filament impinging on a corpuscle, and showing the undulating longitudinal membrane, the long flagellum, find the refractive spherules in the granular protoplasm ( x 3000). the most part slightly tapering bodies which terminate at one end in a stiff, immotile, acutely-pointed flexible filament or spine-like process, and at the opposite end are provided with a long flagellum, while, longitudinally attached, a delicate undulating fin-like membrane can be traced, which starts from the base of the posterior filament, and becomes directly continuous with the flagellum (Fig. 240). With careful illumination the body is found to be distinctly granular, with one or more highly -refractive spherules. When the rapid movement is arrested the undulating membrane is distinctly visible. The best opportunity occurs for seeing this when the organism comes to partial rest with its stiff filament against a corpuscle, as if to obtain a point cFappui, while lashing its flagellum in all directions (Fig. 241, b). At other times, when the parasite has impinged with its posterior extremity against a corpuscle, or the stiff filament is apparently entangled in debris, the movements of the organism give one the idea of its endeavouring to set itself free. 602 APPENDICES. but the author has not been able to persuade himself that they " attack and disintegrate " the red blood- corpuscles. In the active state the thicker portion, or body, appears to twist and bend from side to side with great activity. The organism can turn completely round with lightning rapidity, so that the flagellum, at one moment lashing in one direction, is suddenly observed working in the opposite direction. Then suddenly the organism makes progression, and it can be distinctly seen to move in the direction of the flagellum, the flagellum threading its way between the corpuscles and drawing the rest of the organism after it. Currents set up by evaporation may undoubtedly here and there produce the appearance of the organism "wriggling along" with its flagellum posterior ; but the author was convinced, after FIG. 241.— MONADS IN RAT'S BLOOD, x 1200. a, A monad threading its way among the blood-corpuscles ; b, another with pendulum movement attached to a cor- puscle ; c, angular forms ; d, encysted forms ; e and /, the same seen edgeways. hours of patient observation, that in the normal mode of progression the flagellum acts as a tractellum and not as a pulsellum. By treating cover-glass preparations with osmic acid the appearances obtained are very similar to those shown in Lewis's photographs, so that there is no doubt, in spite of the descriptions not completely according, that they are one and the same organism. There was a great likeness to the organisms described by Mitrophanow, and to the Surra parasite ; and when the author had stained the rat parasites, the closest examination confirmed his belief that they were morpho- logically identical with the stained parasites of Surra. Cover-glasses with a thin layer of blood may be passed three times through the flame of a Bunsen burner in the way commonly employed for examining micro-organisms, and stained with an ANIMAL MICRO-PARASITES. 603 aqueous solution of fuchsin, methyl-violet, or Bismarck- brown, or with aurantia, nigrosin, and other aniline dyes. The following method will, however, be found most instructive: — Use freshly prepared saturated solution of fuchsin or methyl-violet in absolute alcohol, and put a drop with a pipette on the centre of the prepara- tion ; do not disturb the drop-form for a few moments ; then, before the alcohol has evaporated, wash off the excess of stain. It will be found that where the drop rested the organisms will be very deeply stained, while in the surrounding area the colour will vary in intensity. By the effect of the different degrees of staining much FIG. 242.— MONADS IN KAT'S BLOOD STAINED WITH METHYL VIOLET, SHOWING MEMBRANE UNDER DIFFERENT ASPECTS, BLOOD-CORPUSCLES, SOME CRE- NATED AND STAINED DlSCS ( X 1200). may be learnt (Fig. 242). In one organism the body and entire membrane will be equally stained ; in another the margin of the membrane only. In some the posterior stiff filament is stained, and at its base a darkly stained speck is very striking ; and in other cases, again, the posterior filament is only faintly tinged, or an unstained spot occurs near its base. H^EMATOZOA OF FlSH. In the year 1883 Mitrophanow published a paper in which he gave an account of organisms in the blood of the mud-fish and the carp. In the blood of the mud-fish (Cobitis fossilis) the organisms at the first glance looked like minute nematodes, but the appearances, and changes which took place on further examination showed nothing in common with worms (Fig. 243). As a 1 per cent, salt solution had been added to the blood under examination, it occurred 604 APPENDICES. to Mitrophanow that they were possibly the cytozoa described by Gaule ; but this idea was dismissed by the fact that they were found in blood to which 110 salt solution was added. Their size varied from 30 to 40 /* in length and 1 to 1| //, in width. At first their rapid movements baffled examination, but as the rapidity lessened there was the appearance of a curling movement in the body portion and a swinging movement of the lash. The organism moved in the direction of the lash, the anterior end of the body being more pointed than the posterior, and gradually fining off into the lash. When the body seemed to rest, the lash might be seen to FIG. 243.— ORGANISMS IN THE BLOOD OF MUD-FISH (Hfemcitomoncts colitis), a, First Variety ; 6, second variety ; c, third variety, d, First variety in a state of diminished activity, e, The same after treatment with osmic acid. (Mitro- phanow. ) whip out in all directions. As the movement of the body gradually diminished, it appeared to have a complicated screw form, the axis of the screw corresponding to the body to which an undulating membrane is fastened spirally. This could be distinguished when the organism was dying, because the body in death contracted, and the membrane then looked like a spiral addition. Thus the organism consisted of a body, a spiral membrane, and a flagellum. With higher magnification the organism appeared to consist of a refractive, strongly contractile protoplasmic substance, which, when death occurred, formed a shapeless mass. In the same blood two other forms were observed : one without a membrane, but having ANIMAL MICRO-PARASITES. G05 two highly refractive spherules in the protoplasm ; and another with neither membrane nor flagellum, consisting of very granular proto- plasm with several refractive spherules, and capable of protruding processes like pseudopodia. In the carp (Fig. 244) the parasite is perceptibly larger, and possesses an undulating membrane fastened along the edge of the long body. When the body bent first towards one side and then to the other, a wave-like movement was observable at the free edge of this membrane. These parasites were found in all the mud-fish examined except FIG. 244. — ORGANISMS IN THE BLOOD OF THE CARP. a, I, c, Ha'iiMtomonas carassii ; d, e, /, g, h, other organisms in the same blood (Mitrophanow). one, and in greater numbers in the hot months. In the carp tlx-y were only found occasionally. Mitrophanow described other varieties, which he considered were possibly not complete organisms, but developmental forms. He considered that these organisms were infusoria between the genera Cercomonas and Trichomonas, with .luvat similarity to the Trichomonas described in the LieberkulmV glands of fowls and ducks (Eberth). On account of their special habitat, Mitrophanow suggested a new genus — Haematomonas, defining this genus as follow*. :-— Parasites of normal fish-blood, worm-like, actively moving organisms, with indistinct differentiation of body parenchyma. l>odii*> pointed at f,06 APPENDICES. both ends, 30 to 40 p, long and 1 to 1| /* wide. May possess in front a nagellum, and on one side an undulating membrane. Species : — Hsematomonas cobitis. — Body provided with a spiral membrane and a nagellum at the fore-end. Parenchyma of body homogeneous. Second variety, body and nagellum only. Movement undulatory, body containing highly refractive spherules. Third variety, plasma- like body, without membrane or nagellum ; quickly changes form by sending out processes laterally, and contains two to four refractive spherules. Blood of mud-fish. Hsematomonas carassii.— Long bodies, with narrow membrane attached along the whole length; less actively motile. Several forms also observed strikingly smaller than the above ; many disc- shaped. Often seen attached to a red corpuscle, setting them in motion by their movements. Blood of carp. The morphological identity of the rat and Surra parasites has been established by the author, and both saem morphologically identical with the organism of Mitrophanow. If we follow Mitrophanow, we must obviously enlarge his genus of Hsematomonas. The author doss not agree with Mitrophanow in the advisability of adopting this entirely new generic name. Mitrophanow suggested this new term because of the special habitat, normal fish-blood, of the species he discovered. But the characteristic features of these organisms are the characteristic marks of the genus Trichomonas. It seems, therefore, that they are embraced by the old genus Trichomonas, and that there is no need to create a new one — Hsematomonas. The common habitat of these species may be expressed by grouping them together in one sub-genus — Trichomonas sanguinis ; but the question arises whether they are distinct species. If it were not for the different description given by Mitrophanow of the organism in the mud-fish, the author would be inclined to say that all these organisms belonged to one and the same species, which might well be named Trichomonas sanguinis. The monad in the rat and the Surra parasite are morphologically identical with each other, and both, as far as one can judge from the description, morphologically identical with the monad in the blood of the carp. We have, however, seen that the organism in Surra is believed to be pathogenic, and too much stress must not be laid on morphological identity. There is strong evidence in favour of believing in its pathogenic properties; but at the same time it must be borne in mind that the organism has never been isolated apart from the blood, and the disease then produced by its introduc- tion into healthy animals. It is quite possible that the parasites in ANIMAL MICRO-PARASITES. 607 Surra are only associated with the disease, the impoverished blood affording a suitable nidus for their development, while the con- taminated water may be the common source of the organism and of the disease. On the other hand, the organism in the rat is found in apparently perfectly healthy, well-nourished animals. The author suggests that the parasites observed in the rat and hamster should be named after Lewis, Trichomonas Lewisi', the organism in the mule, camel and horse after its discoverer, Trichomonas Evansi', and that the names Trichomonas cobitis and Trichomonas carassii should be substituted for the names of the species described by Mitrophanow. Thus we should have added provisionally to the Genus— TRICHOMONAS. Sub-genus — Trichomonas sanguinis. Definition : Elongated tapering bodies, provided with a spiral (T. cobitis), or longitudinal (T. carassii, Lewisi, Evansi) membrane, ter- minating in a rigid filament and an anterior flagellum. Highly polymorphic. Habitat, the blood. Species. — Trichomonas cobitis (Hcem,atomonas cobitis Mitrophanow) — Mud-fish. Trichomonas carassii (tfcematomonas carassii Mitrophanow) — Carp. Trichomonas Lewisi (Herpetomonas Lewisi Kent) — Rat, hamster. Trichomonas Evansi — (Spirochceta Evansi Steel) — Horse, mule, camel ; (pathogenic ?). H^MATOZOA OF THE FROG. Lankester described an organism which he had discovered in the blood of the frog (Rana esculenta). It consisted of a minute pyri- form sac, with the narrower end bent round on itself somewhat spirally, and the broader end spread out into a thin membrane, which exhibited four or five folds and was prolonged on one side into a very long flagellum. The wall of the sac was striated, nucleated and granular ; the membrane undulated during life, and the flagellum was also motile. It was named Undulina ranarum, but subsequently recognised as idnetical with Tnjpanosoma sanguinis described by Gruby. In the same blood Lankester also discovered little oblong bodies, in many cases attached to the end of the red corpuscles, and suggested a geneticai connection with the Undulina. 608 APPENDICES. One or more motionless filaments were occasionally observed attached to these bodies. Gaule subsequently observed the same bodies, and regarded them as resulting from the metamorphosis of the cells of the frog's blood. Gaule's observations were refuted by Lankester in 1882, the parasitic nature insisted upon, and the organism named Drepanidium ranarum. Lankester suggested that they were probably the young stage of a sporozoon allied to Sarcocystis or to Coccidium APPENDIX III. PSOROSPERMS OR COCCIDI A.— AMOEBA COLI. PSOROSPERMS OR COCCIDIA. GREYISH- WHITE nodules may occasionally be found in the liver of a rabbit, the result of a disease which may be mistaken for tuber- culosis. This disease often proves fatal, and may occur in an epidemic form in rabbit warrens. The nodules have cheesy or purulent contents, which are found, on microscopical examination, to contain great quantities of Coccidium oviforme. The coccidia pass from the intestine into the bile-ducts. The walls of the bile -ducts become dilated and folded ; and irregular cavities result from the partial or complete disappearance of the dividing walls of the altered ducts. The folds are composed of connective tissues lined with columnar epithelium, and the coccidia, in different stages of development, are found between the cells, and free in the cavities of the nodules. The individual coccidia are egg-shaped bodies. They possess a thick smooth shell, with an opening, or micropyle, at one end, and protoplasmic contents which may completely fill the capsule or be collected into a spherical mass. After passing from the liver and intestine, -these oval bodies undergo a further development. According to Leuckart, who has very fully described this parasite, the protoplasmic contents divide into four masses, and from each is developed a C-shaped hyaline rod, the cavity of which is occupied by closely packed granules. In this condition they remain until they gain access to a fresh host. Coccidium oviforme has been found in the human liver, and also in sheep, dogs, and cats. Similar, but not identical, bodies occur in mice, and also in fish and other cold-blooded animals. Miescher's tubes are peculiar structures found in swine, cattle, sheep, deer, and mice. They consist of a firm envelope inclosing a number of reniform or bean-shaped bodies. 609 39 610 APPENDICES. Pfeiffer's bodies.— Pfeiffer has described certain appearances which he attributes to coccidia, in epithelial cells in small-pox, vaccinia, and other vesicular diseases. They are probably only derived from the cell nucleus, and are not parasites. Canc&r bodies. — In sections of malignant growths stained by aniline dyes, certain bodies have been found and minutely described and figured by various investigators, and a causal relation sug- gested. Darier first described bodies like cysts, with spores, in Paget's disease of the nipple. Wickham found similar structures and figured them. Nils Sjobring described a cancer parasite, and illustrated his researches with plates. Russell drew attention to certain bodies in cancerous tumours, with a great affinity for fuchsine. Soudakewitch, Podwyssozki, Sawtschenko, Ruffer, and Walker have, among others, contributed to the literature of the so- called cancer parasites. These bodies appear in the form of ref ractile spherical elements, which stain well with reagents, such as the Ehrlich-Biondi stain. Sections are left in this stain for twenty-four hours, washed in alcohol, cleared in xylol, and mounted in xylol balsam. The spherical bodies have sometimes a radiate appearance. These bodies have not been cultivated, and inoculation experiments with cancerous tissue have been negative. The opinion is now very generally held that these bodies are not parasites, but that changes occur in the cells and nuclei, resulting in the formation of peculiar structures, which have been brought to light by the use of aniline dyes and complex staining methods. We are justified in concluding that the cause of cancer is unknown. Ballance and Shattock have made repeated attempts to cultivate parasitic protozoa from malignant tumours, and they have extended their researches to vaccinia and molluscum contagiosum, but with negative results. Sand and water were used as the medium for these experiments. Cultivations were made from nine scirrhous carcinomata of the breast, five sarcomata from different sources, two melanotic sarcomata from horses, and a sarcoma from a dog. In every instance the result was negative. No traces of protozoic life could be found, in spite of examinations at regular intervals, and repeated for periods of many months. AMCEBA COLI. Lbsch, Grassi, Kartulis, and others have described an amoeba in the intestines of patients suffering from dysentery. Losch adminis- tered the fresh dejecta of a patient containing the amoeba? to dogs, ANIMAL MICRO-PARASITES. 611 and in one case a mucous mass was passed containing a number of amoebae. Eighteen days afterwards the dog was killed, and the mucous membrane of the intestine was reddened, swollen, and m "(0 FIG. 245.— AMCEBA COLI IN INTESTINAL Mucus (Loscn). ulcerated in three places. The mucus in the rectum and in the ulcers contained numerous amoebae. Cunningham, who has found the amoebae in choleraic and other cases, and in the intestine of the cow and horse, does not attach any importance to their presence. APPENDIX IV. APPARATUS, MATERIAL, AND REAGENTS EMPLOYED IN A BACTERIOLOGICAL LABORATORY. (A) HlSTOLOGICAL APPARATUS. Microscope. — For the investigation of micro-organisms a good microscope with oil-immersion system and a condenser, such as Abbe's, is essential. Such instruments are supplied by Zeiss, Leitz, Reichert & Hartnack in Germany, and Powell & Lealand, Swift & Baker in England. Zeiss supplies a micrometer eyepiece, with -directions for use. Some such arrangement is essential for the measurement of bacteria. Other accessories to the microscope are : A large bell-glass, for covering the microscope when not in use. About a foot square of blackened plate-glass. A white porcelain slab of the same size. Glass bottles, with ground -glass stoppers, for alcoholic solutions of aniline dyes, etc. Glass bottles, with funnels, for aqueous solutions of the dyes, and -others provided with pipettes. A small rod-stoppered bottle of cedar oil. This is recommended by Zeiss in preference to other oils for his immersion lenses. Set of small glass dishes or capsules and watch-glasses, for section- staining, etc. Stock of best glass slides, in packets of fifty. Several boxes of round and square thin cover-glasses, in various sizes, of the best quality. Needle-holders, with a couple of platinum needles, and a packet of ordinary sewing-needles. Glass rods drawn out to a fine point ; useful for manipulating sections when acids are employed. Platinum or plated copper section-lifters. One pair of small brass or spring-steel platinum-pointed forceps, for holding cover-glasses. One pair of brass tongg. 612 APPARATUS, MATERIAL, AND REAGENTS. 61$ Collapsible tubes, for containing Canada balsam ; very serviceable for transport and general use. Turn-table for sealing cover-glass preparations, with rings of cement. Boxes for preparations, book-form. Tickets and labels, various sizes. Soft rags or old pocket-handkerchiefs, for removing cedar oil from immersion lens, cleaning cover-glasses, etc. Chamois leather for wiping lenses. Warm Stages. — In addition to those already described, Schafer and Strieker have constructed warm stages for accurate observations. Schafer's apparatus consists of a vessel (/), filled with water which has been boiled to expel the air, and heated by means of a gas-flame at g. The warmed water ascends the indiarubber tube (c) to the FIG. 246.— SCHAFER'S WARM STAGE. brass box (a). The box is pierced by a tubular aperture to admit light to the object, and has an exit tube (c'), by which the cooled water from the stage returns to be reheated by the flame g. At d is a gas-regulator, so that a constant temperature at any desired point can be maintained. Strieker's stage, in which warm water or steam can be used for heating, and by the employment of iced water also used for observing the effects of cold, is shown in Fig. 247. It consists of a hollow rectangular box, with a central opening (C) permitting the passage of light. The water makes its exit and entrance at the side tubes (a, a), and the temperature is indicated by a thermometer in front. 614 APPENDICES. A more complicated apparatus, combining both a warm stage and a gas chamber, is shown in Fig. 248. This consists of a rect- angular piece of ebonite (E E) fixed to a brass plate which rests on the stage of the microscope. On the upper surface of the ebonite is'another brass plate (P), with an aperture (G) leading into a brass FIG. 247.— STEICKEK'S WARM STAGE. tube closed below by a piece of glass. To heat the apparatus the copper wire B is placed on the tube a, and its extremity heated by the flame of the lamp. The nearer the lamp to the stage the higher the temperature, which is indicated by the thermometer (t). To J> FIG. 248.— STRICKEB'S COMBINED GAS CHAMBER AND WARM STAGE. employ it as a gas chamber the wire B is laid aside, and the gas is conducted into the chamber by the tube a', and escapes by the tube a. Microtome — Schanze's is much in favour in Germany, but Jung's of Heidelberg, though a somewhat cumbrous instrument, is APPARATUS, MATERIAL, AND REAGENTS. 615 preferred by many workers. Smaller accessories, which should be within reach, are — A small can of sewing-machine oil. A soft rag and chamois leather, for wiping the knives immediately after use. Stone and leather, for setting and sharpening the same. Two or three camel's-hair brushes. A freezing microtome is very useful : such as Swift's, which is used by the author; and the method of embedding in celloidin is combined with the ordinary process of freezing. (B) REAGENTS AND MATERIAL EMPLOYED IN THE PROCESSES or HARDENING, DECALCIFYING, EMBEDDING, FIXING AND CUTTING OF TISSUES. Alcohol, absolute. Bergamot oil. Celloidin. Dissolved in equal parts of ether and alcohol. Cork, or stock of ready-cut corks. Ebner's solution. A mixture in the following proportions : — Hydrochloric acid .... '5 Alcohol ,100 Distilled water 20 Chloride of sodium ... 5 Formalin. Gelatine. Melted in a small porcelain capsule, and set aside ready to be re-melted when required for use. Glycerine gelatine (Klebs). Best well- washed gelatine . . . .10 Add distilled water, allow gelatine to swell up, pour off excess of water, melt gelatine with gentle heat, add Glycerine ..... .10 Lastly, a few drops of phenol for preservation. Gum. 616 APPENDICES. Kleinenberg's solution. Saturated watery solution of picric acid . .100 Strong sulphuric acid ... . . . 2 Filter, and add Distilled water 300 Miiller's fluid. Bichromate of potash . . . ... 2 Sulphate of sodium . . . . . . 1 Distilled water . . ... . ... 100 Osmic acid. Distilled water . . •....« -• • .100 Osmic acid . . . . . . . '. . -5 Paper trays (or small glass capsules). Paraffine. Spermaceti. Xylol. (C) REAGENTS FOR EXAMINING AND STAINING MICROSCOPICAL PREPARATIONS. 1. Acetic acid, strong. 2. Alcohol, absolute. 3. Alcohol, 60 per cent. 4. Alcohol, acidulated. Alcohol ..... 100 Hydrochloric acid ... 1 5. Alum Carmine (Grenadier). Carmine . . . j Five per cent, solution of alum . . .100 Boil twenty minutes,; filter when cold. 6. Ammonia, strong. 7. Aniline. 8. Aniline water. Distilled water . 100 Aniline . K • • . • . • •• v1 ... y Shake well, and filter emulsion. APPARATUS, MATERIAL, AND REAGENTS. 617 9. Bismarck-brown. (a) Concentrated solution in equal parts of glycerine and water. (b) Aqueous solution. Bismarck-brown . . . . . 2 Alcohol . . . .". . . . .15 Distilled water 85 10. Borax-carmine (Grenacher). Borax 2 Carmine ........ 5 Distilled water . . . . . .100 To the dark purple solution add a 5 per cent, solution of acetic acid until a red colour is produced ; set aside twenty-four hours ; filter, and add a drop of carbolic acid. 11. Cedar oil. 12. Ehrlich-Biondi solution (Heidenhain). To Saturated aqueous solution of Orange. G. .100 Add Saturated aqueous solution of Rubin. S. . . 20 „ „ „ Methyl-green. OO. 50 To the mixture .... 1 Add water 100 13. Eosin. (a) Saturated alcoholic solution. (b) Aqueous solution. Distilled water. .... .100 Eosin . . .... 5 14. Ether. 15. Fuchsine. (a) Saturated alcoholic solution. (b) Aqueous solution. Fuchsine .... Alcohol .15 Water ..... .85 16. Gentian- violet. (a) Saturated alcoholic solution. (6) Aqueous solution. Gentian-violet. . . 2 -25 Distilled water . 100 OP THH 1 UNIVERSITY 618 APPENDICES. 17. Gibbes' solution, for double staining. Take of Rosaniline hydrochlorate ..... 2 Methylene-blue ...... 1 Triturate in a glass mortar. Dissolve aniline oil . . . . . 3 In rectified spirit ... . . . .15 and add slowly to the above. Lastly, slowly add distilled water . . .15 Keep in stoppered bottle. 18. Glycerine, pure. 19. Hsematoxylin solution. Hsematoxylin . . . . . . . 2 Alcohol 100 Distilled water . . . . . .100 Glycerine ....... 100 Alum ........ 2 20. Iodine solution. Iodine, pure ....... 1 Iodide of potassium ...... 2 Distilled water 50 21. Iodine solution (Gram). Iodine ..... 1 Iodide of potassium . . . . . 2 Distilled water 300 22. Lithium-carmine solution (Orth). Saturated solution of carbonate of lithium . 100 Carmine ...... . 2'5 23. Magenta solution (Gibbes). Magenta ...... 2 Aniline oil ... . 3 Alcohol (sp. gr. -830) 20 Distilled water .... 20 24. Methylene-blue. (a) Concentrated alcoholic solution. {b) Aqueous solution. Methylene-blue .... 2 Alcohol 15 Water . . 85 APPARATUS, MATERIAL, AND REAGENTS. 619 •(c) Koch's solution. Concentrated alcoholic solution of methylene-blue 1 Ten per cent, potash solution .... Distilled water 200 (d) Loffler's solution. Concentrated alcoholic solution of methylene-blue 30 Solution of potash, 1 to 10,000 . . . 100 25. Methyl-violet. (a) Concentrated alcoholic solution. (b) Aqueous solution. Methyl- violet . . . . . . . 2 -2 5 Distilled water ... . . .100 (c) Koch's solution. Aniline water . . . . . . .100 Alcoholic solution of methyl-violet . .11 Absolute alcohol . . . . . .10 26. Neelsen's solution. Dissolve f uchsine ..... In alcohol 10 Add a 5 per cent, watery solution of carbolic acid 100 27. Nitric acid, pure. 28. Orseille (Wedl). Dissolve pure ammonia-free orseille in Absolute alcohol ... .20 Acetic acid .... 5 Distilled water . • 40 until a dark red liquid results. Filter. 29. Picric acid. (a) Concentrated alcoholic solution. (b) Saturated aqueous solution. 30. Picro-carmine (Ranvier). Carmine . Distilled water . .10 Solution of ammonia 3 Triturate ; add cold saturated solution of picric acid .... 200 620 APPENDICES. 31. Picro-lithium-carmine (Orth). To above-mentioned lithium-carmine solution add saturated solution of picric acid . . 2'3 32. Potash solution. (a) 1 to 3 per cent. (b) 10 „ „ (c) 33 „ „ 33. Safranine. (a) Concentrated alcoholic solution. (b) Watery solution . . . . ; 1 per cent. 34. Sulphuric acid, pure. 35. Salt solution . : .'.. ,' . ,' . . 0-8 per cent. 36. Turpentine. 37. Vesuvin. (a) Concentrated alcoholic solution. (b) Watery solution. Water, distilled. Water, sterilised. Distilled water can be kept for use in a wash bottle, or far better in a siphon apparatus. Sterilised water is convenient in plugged sterile test-tubes, which may be kept close at hand in a beaker, or tumbler, with a pad of cotton wool at the bottom. The numbered reagents can be conveniently arranged on shelves within easy reach. Alcoholic solutions of the aniline dyes and other special preparations should be kept in bottles with ground- glass stoppers. Aqueous solutions of the dyes may be kept in bottles with funnel niters, and the solution filtered before use. To both aqueous and alcoholic solutions a few drops of phenol, or a crystal of thymol, should be added as a preservative. For the rapid staining of cover- glass preparations, it is convenient also to have the most frequently used stains (fuchsine, methyl- violet) in bottles provided with pipette stoppers. (D) REAGENTS FOR MOUNTING AND PRESERVING PREPARATIONS. Acetate of potash. Concentrated solution. Asphalte lac. APPARATUS, MATERIAL, AND REAGENTS. 621 Canada balsam. Dissolved in xylol. Glycerine gum (Farrant's solution). Glycerine. Water. Saturated solution of arsenious acid. Equal parts ; mix, and add of picked gum arabic half a part. Hollis' glue. Zinc-white. (E) DRAWING AND PHOTOGRAPHIC APPARATUS. Camera Lucida. — The camera lucida of Zeiss is an excellent instrument, though many prefer the pattern made by Nachet of Paris. Combined with the use of a micromillimeter objective, it affords also a simple method for the measurement of bacteria. For drawing microscopical appearances, and for illustrating microscopical specimens with or without the use of a camera lucida, the following materials should be within reach : — Pencils. Etching pens. Prepared Indian ink. Water-colour paints and brushes. Ordinary and tinted drawing paper and other usual accessories. Photo-micrographic Apparatus, — Zeiss of Jena, Seibert & Kraft of Wetzlar, Nachet of Paris, and Swift & Son of London, may all be recommended for constructing an arrangement in which the photographic camera is combined with the microscope. The best models have been described fully in the chapter on Photography of Bacteria. The accompanying figure (Fig. 249) illustrates a model in which the microscope is used in the vertical position. For illumination either sunlight or artificial light may be em- ployed. In the case of sunlight a heliostat is necessary to procure the best results ; but as sunlight is not always available by day, and it is also more convenient for many to work at night, it is better to have recourse altogether to artificial light. Excellent results may be obtained with an ordinary paraffine lamp, or with magnesium, oxycalcium, or electric light. 622 APPENDICES. FIG. 249.— VEKTICAL MICRO-PHOTOGKAPHIC APPARATUS. (F) STERILISATION APPARATUS. Steam-steriliser. — A cylindrical vessel of tin about half a metre or more in height, jacketed with thick felt, and provided with a conical cap or lid (Fig. 250). The lid is also covered with felt, has handles on either side, and is perforated at the apex, to receive a thermometer. Inside the vessel is an iron grating or diaphragm about two-thirds the way down, which divides the interior into two chambers — the upper or " steam-chamber," and the lower or "water-chamber." A gauge outside marks the level of the water in the lower chamber ; this should be kept about two-thirds full. APPARATUS, MATERIAL, AND REAGENTS. The apparatus stands upon three legs, and is heated from below with two or three Bunsen burners, or a Fletcher's burner. It is employed for sterilising nutrient media in tubes or flasks, for cooking potatoes, or hastening the filtration of agar- agar. When the thermometer indicates 100° C. the lid is removed, and test-tubes are lowered in a wire basket by means of a hook and string, and the lid quickly re- placed. Potatoes or small flasks are lowered into the cylinder in a tin receiver with a perforated bottom, which rests upon the- grating and admits of its contents being exposed to the steam. A larger model is shown in Fig. 33. Hot-air Steriliser. — A cubical chest of sheet iron with double walls, supported on four legs ; it may also be suspended on the wall of the laboratory, with a sheet of FiG.250.-KocH's STEAM- Asbestos intervening (Figs. 251 and 252). It is heated with a rose gas-burner from below, and the temperature of the interior in- STERILISEE. clicated by a thermometer inserted through a hole in the roof ; in a second opening a gas regulator can be fixed. Test-tubes, flasks, funnels, cotton wool, etc., may be sterilised by exposure to a temperature of 150° C. for an hour or more. FIG. 251.— HOT-AIR STERILISER. FIG. 252.— SECTION OF HOT- AIR STERILISER. 624 APPENDICES. (G) APPARATUS AND MATERIAL FOR PREPARING AND STORING NUTRIENT GELATINE AND NUTRIENT AGAR-AGAR. Water-bath. — A water-bath on tripod stand is required for boiling the ingredients of nutrient jellies and for general purposes. The lid may be conveniently composed of a series of concentric rings, so that the mouth of the vessel may be graduated to any size required. Test-tube Water- bath. — This consists of a circular rack for test-tubes within a water-bath. It is sometimes employed instead of the steam cylinder for sterilising nutrient jelly in tubes, by boiling for an hour for three successive days. Hot- water Filter. — A copper funnel with double walls, the inter- space between which is filled "with hot water. A glass funnel fits in- side the copper cone, the stem of the glass funnel passing through and being tightly gripped by a per- forated caoutchouc plug, which fits in the opening at the apex of the cone. The water in the cone is heated by applying the flame of a burner to a tabular prolongation of the water- chamber. In a more recent model, as represented in Fig. 31, this prolongation is dis- pensed with, and the temperature is maintained by means of a circular burner which acts at the same time as a funnel ring. In Rohrbeck's model the funnel of the filter is connected with a flask, from which the test-tubes can be easily filled with the liquid jelly (Fig. 253). Glass Vessels.— A number of glass vessels should be kept in stock according to requirements. Bohemian hard glass flasks are employed in several sizes, for boiling nutrient media. The conical forms are especially used in the larger sizes for storing nutrient jelly. Glass funnels, large and small, are necessary, not only in the processes of preparing nutrient jelly, but for filtering solutions of aniline dyes and for general purposes. FIG. 253.— HOT- WATER FILTERING APPARATUS WITH RING BURNE R. APPARATUS, MATERIAL, AND REAGENTS. 625 A liberal supply of test-tubes should always be kept in stock, as they are not only employed for the tube- cultivations, but can be conveniently used for storing bouillon, sterilised water, etc. Cylindrical glasses graduated in cubic centimetres, 10 ccm., 100 ccm., 500 ccm., are required for measuring the liquid ingredients of nutrient jelly, and also in preparing the various staining .solutions. A large wide- mouthed glass jar, with a glass cover, is extremely useful. It must be padded at .the bottom with cotton wool for containing a stock of tubes of sterilised nutrient jelly, and should be placed within reach on the working table. Balance and Weights.— A balance, with large pans and set of gramme weights, is constantly required. Cotton Wool. — The best or " medicated " cotton wool should be procured. , Gelatine. — The gelatine for bacteriological purposes must be of the very best quality (gold label). Agar-agar. — This is also called Japanese Isinglass ; it consists of the shrivelled filaments of certain Algse (Gracilaria lichenoides and Gigartina speciosa). Peptonum Siccum. Table Salt. — Prepared table salt can be obtained in tins or packets. Litmus Papers. — Blue or red litmus paper in cheque-books, for testing the gelatine mixture, etc. Carbonate of Soda. — A bottle, containing a saturated solution of carbonate of soda, and provided with a pipette stopper, may be kept, especially for use in the preparation of nutrient jelly. Lactic Acid. Filter Paper. — For filtering gelatine, stout Swedish filter paper of the best quality is recommended. Flannel or Frieze. — This is employed as a substitute for, or combined with, filter paper in the preparation of nutrient agar- agar. (H) APPARATUS FOR EMPLOYMENT OF NUTRIENT JELLY IN TEST-TUBE AND PLATE-CULTIVATIONS. Wire Cages. — These cages or crates are used for containing test-tubes, especially when they are to be sterilised in the hot-air steriliser ; or for lowering tubes of nutrient jelly into the steam- steriliser, etc. (Fig. 254). 40 626 APPENDICES. Test-tube Stands.— The ordinary wooden pattern, or the metallic folding stands, are called into use for holding cultivations. Pegged racks are also recommended for draining test-tubes after washing. Caoutchouc Caps. — These are caps for fitting over the cotton -wool plugs, and may be used in different sizes for test-tubes and stock-flasks. Platinum Needles. — A platinum needle for inoculating nutrient media, ex- amining cultivations, etc., consists of two or three inches of platinum wire fixed to the end of a glass rod. Several of these needles should be made with platinum wire of various thicknesses. A piece of glass rod, about seven inches long, is heated at the extreme point in the flame of FIG. 254.— WIRE CAGE FOB TEST-TUBES. FIG. 255. — PLATINUM NEEDLES; STRAIGHT, HOOKED, LOOPED. a Bunsen burner, and a piece of platinum wire, held near one extremity with forceps, is then fused into the end of the rod. Some needles should be perfectly straight, and kept especially for FIG. 256.— DAMP CHAMBER FOR PLATE-CULTIVATIONS. inoculating test-tubes of nutrient jelly. For other purposes the needles may be bent at the extremity into a small hook, and )thers provided with a loop (Fig. 255). APPARATUS, MATERIAL, AND REAGENTS. 627 Tripod Levelling-stand. — A triangular wooden frame sup- ported upon three screw-feet which enable it to be raised or lowered to adjust the level. FIG. 257. — APPARATUS EMPLOYED FOR PLATE-CULTIVATIONS. Tripod Stand ; Glass Dish, filled with cold or iced water ; Sheet of Plate-glass ; Spirit Level, and Glass Bell. Large Glass Plate. — A piece of plate-glass, or a pane of ordinary window-glass, about a foot square. Spirit Level. Glass Bells and Dishes. — Shallow glass bells and dishes, for making a dozen or more damp chambers (Fig. 256), and for completing the apparatus for pouring out liquefied nutrient jelly on glass plates or slides (Fig. 257). Iron Box A box of sheet-iron (Fig. 258), for containing glass plates during their sterilisation in the hot-air steriliser, and for storing them until required for use. Glass Plates. — Small panes of glass, about six inches by four. Not less than three dozen are required for a dozen damp chambers. Glass Benches. — These are necessary for arranging the glass- plates or slides in tiers in the damp chambers (Fig. 256). Metal FIG. 258.— Box FOR GLASS PLATES. FIG. 259.— GLASS BENCHES FOR GLASS PLATES OR SLIDES. shelves may be substituted for them, but the former are to be preferred. They can be easily made, in any number required, by APPENDICES. cementing a little piece of plate-glass at either end of a glass .slip (Fig. 259). Glass Rods. — One dozen or more glass rods, twelve to eighteen inches in length. They are employed for smoothly spreading out the liquefied nutrient gelatine or agar-agar on the glass plates, etc. Thermometers. — Two or three centigrade thermometers. (I) APPARATUS FOR PREPARATION OF POTATO-CULTIVATIONS. Israel's Case. — Sterilising instruments in the flame of a Bunsen .burner is most destructive. It is better, therefore, to have a sheet- iron case (Fig. 260) to contain potato-knives, scalpels and other in- struments, and to ster- ilise them by placing the case in the hot-air steriliser for an hour at 150° C. The box can be opened at the FIG. 260.— ISRAEL'S CASE. side, and each instru- ment withdrawn with a pair of sterilised forceps when required for ase Glass Dishes.— Several shallow glass dishes are required for preparing damp chambers for potato-cultivations (Fig. 261). The upper, being the larger, fits over the lower, and having no handle, admits of these damp chambers being placed, if necessary, in the incubator in tiers. The large size may also be used in the same way for plate-cultivations. Potato Knives. — A common broad smooth-bladed knife set in a wooden handle is sold for this purpose. Scalpels.— Half a dozen scalpels, preferably with metal handles, may be kept especially for inoculating sterilised potatoes. Brush.— A common stout nail-brush, or small scrubbing-brush, is essential for cleansing potatoes. FIG. 261.— DAMP CHAMBER FOR POTATO- CULTIVATION. APPARATUS, MATERIAL, AND REAGENTS. (J) APPARATUS FOR PREPARATION OF SOLIDIFIED STERILE BLOOD-SERUM. Glass Jar. — A tall cylindrical glass jar, on foot, with a broad ground stopper, for receiving blood. Pipette. — An ordinary or graduated pipette, for transferring the- seruin from the jars to sterile test-tubes or glass capsules. Koch's Serum Ster- iliser.—A cylindrical case, with double walls forming an interspace to contain water, closed with a lid, also double- walled and provided with a tubular prolongation of the enclosed water-chamber (Fig. 262). The water in the cylinder is heated from below, and that in the lid by means of the prolongation. In the centre of the cylinder is a column which communicates with the water- charnber of the cylinder, and from it pass four partitions, which serve to support the test -tubes. In the lid are three openings, one of which communicates with the water-chamber in the lid by which the latter is filled, and into which a thermometer is then fixed. In the centre an opening admits a thermo- meter, which passes into the central pipe of the cylinder ; through a third opening a thermometer passes to the cavity of the cylinder. The cylinder and cover are jacketed with felt, and the apparatus is supported on iron legs. FIG. 263. -SERUM INSPISSATOR. Koch's Serum Inspis- sator. — A shallow tin case with glass cover, both case and cover jjucketed with felt (Fig. 263) FIG. 262.— KOCH'S SERUM STERILISER. . Finkler and Prior, Naturforscherversammlung Magde- burg, 1884; Deut. Med. Woch., Nr. 36, 1884; Erganzungshefte zum Centralbl. f. Allg. Gesundheitspflege, Bd. I., Heft 5 u. 6, 1885. Fliigge, Deut. Med. Woch., Nr. 2, 1885. Gaffky, Arb., aus d. K. Gesundheitsamte, 1887. Gamaleia, Compt. Rend., 1888; Compt. Rend. Soc. de Biolog., 1889. Gibier and van Ermengem, Compt. Rend., T. 101, 1885. Haffkine, Brit. Med. Journ., 1895. Hericourt, Compt. Rend., 1885. Hueppe, Berl. Klin. Woch., 1887 ; Deutsche Med. Woch., 1887; Compt. Rend., 1889; Prag. Med. Woch., 1889. Hunter, Brit. Med. Journ., 1884. Johne, Deutsche Zeitschr. f. Thiermed., Bd. XI., 1885. Kitasato, Zeitschrif,t f. Hygiene, 1888, 1889. Klebs, Ueber Cholera Asiatica, 1885. Klein, Brit. Med. Journ. and Proc. Roy. Soc. London, No. 38, 1885 ; Bacteria of Asiatic Cholera, 1889. Klein and Gibbes, An Inquiry into the Etiology of Asiatic Cholera : Bluebook, 1885. Koch, Deut. Med. Woch., 1884 ; Etiology of Cholera : Berlin Cholera Conference : Translated by Laycock, in Microparasites and Disease (New Syd. Soc.), 1885. Lewis, Med. Times and Gazette, 1884. Lustig, Centralbl. f. die Med. Wiss., 1887 ; Zeitsshrift f. Hygiene, 1887. Macnamara, Brit. Med. Journ., 1884. Miller, Deut. Med. Woch., Nr. 9, 1885. Neuhaus, Centralbl. f. Bacteriolog., 1889. Nicati and Rietsch, Arch, de Physiol., 1885: Revue de Medecin, T. 5, 1885; Revue d'Hygiene, 1885. Petri, Centralbl. f. Bacteriolog., 1889. Pettenkofer, Lancet, 1886. Pfeiffer, Corresp.-Bl. des Allgem. Aerztl. Ver. inThiiringen, Nr. 9, 1884 ; Deut. Med. Woch., 1886-8S. Pfuhl, Zeitschr. f. Hygiene, 1889. Boy, Brown jtad Sherrington, Proc. Roy. Soc., Vol. xli. Salkowski, Virchow's Arc'aiv, 1887. Schottelius, Deut, Med. Woch., Nr. 14, 18S5. Strauss, EDUX, Thuillier and Nocard, Compt. Rend. Soc. de Biol., T. 4, 1883. 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Derm, und Syph., 1887. Lustgarten and Manneberg, Vierteljahresber. fiir Dermat. und Syph., 1887. Lustig, Diagnostik der Bakt. d. Wassers. Mace", Traite pratique der Bacteriolog.. 1892. Malerba, Giorn. Intern, d. Sci. Med., 1888. Manfredi, Fortschr. der Med., 1886. Manneberg, Centralb. f. Klin. Med., 1888. Marpmann, Erganzungshefte des Centr. f. Allgem. Gesund., Bd. II. Maurea, Centralbl. f. Bakteriol., Bd. XL, 1892. Mendoza, Centralbl. f. Bakteriol., Bd. VI. Menge, Centralbl. f. Bakteriol., Bd. VI., 1889 ; Bd. XII., 1892. Miller, Deutsch. Med. Woch., 1884, 1886, 1887 ; Micro-organisms of the Human Mouth. 1890. Miquel, Ann. de Micrographie, 1888-92. Mori, Zeitschr. f. Hygiene, Bd. IV., 1888. Mulhauser, Virch. Arch., Bd. 97, 1884. 664 APPENDICES. Hunk, Virch. Arcb., Bd. 22, 1861 ; Med. Centralbl., 18G4. Muntz, Compt. Rend.! T. cxii. Neelsen, Beitrag. z. Biol. der Pttanzen, III. Neisser, Archiv f. Hygiene. 1893. Neumann and Schaeffer, Virchow's Archiv, Bd. CIX., 1887. Nocard, Ann. de I'lnstitut Pasteur, 1887. Nocard and Mollereau, Ann. de I'lnstitut Pasteur, T. 1, 1887. Oersted, Naturhist. Tidsskrift, Bd. III., 1840. Okada, Centralbl. f. Bacteriolog., Bd. IX., 1891. Paltauf and Heider, Med. Jahrbuch., 1889. Passet, Zeitschr. f. Hygiene, Bd. IX. ; Untersuch. u. die eitr. Phleg. des Wiensch., 1885 ; Fortschr. der Med., 1888. Pasteur, Ann. de Chim. et de Phys., T. 64, 1862 ; Compt. Rend., 1876; LIL, 1861. Pansini, Virch. Archiv, Bd. cxxii.. 1890. Perdrix, Ann. de 1'Institut Pasteur, T. v., 1891. Perty, Zur Kentniss. Kleinst. Lebensform, 1852. Pfeiffer, Deutsche Med. Woch., 1888 : U. die Bac. Pseudotuberculose bei Nagethiere, 1889 ; Zeitschr. f. Hygiene, Bd. VI. ; Bd. VIII., 1889. Plagge and Proskauer, Zeitsch. f . Hygiene, II. Pohl, Centralb. f. Bacteriolog., 1892. Pommer, Mitth. des Bot. Inst. zu Gratz., I. Popoff, Ann. de 1'Institut Pasteur, 1890. Prazmowski, Untersuch. ii. die Entwick. und Fermentwirkung einer Bakterien, 1880. Prove, Beitr. z. Biolog. der Pflanzen, Bd. III. Raczynsky, Diss. der Militar. Med. Akad., 1888. Keimann, Inaug. Diss. Wurzburg, 1887. Renon. Ann. de 1'Institut Pasteur, 1892. Roscoe and Lunt, Phil. Trans. Roy. Hoc., 1892. Rosenthal, Zeitschr. f. Hygiene, V. Roth, Zeitschr. f. Hygiene, Bd. VIII., 1890. Russell, Zeitschr. f. Hygiene, Bd. XL, 1891. Sakharoff, Ann. de I'lnstitut Pasteur, 1891. Sanarelli, Centralbl. Bd. IX. 1891. Scheibenzuber, Allgem. Wien. Med. Zeit., 1889. Scheurlen, Deutsche Med. Woch, 1888. Schimmelbusch, Deut, Med. Woch., 1889. Scholl, Fortschr. der Med., VII. Schottelius, Biol. Untersuch. ii. dem Micrococcus Prodig., 1887 ; Zeitschr. f. Hygiene, 1890. Schrb'ter, Beitrage z. Biol. d. Pflanz., Bd. 1, H. 2. Schutz, Archiv f. Wiss. und Prakt. Thierheilk., 1888. Senger, Berl. Klin. Woch., 1888. Smith, Central, f. Bakteriolog., Bd. X. ; Med. News, 1887. Sternberg, Report on Etiology and Prev. of Yellow Fever, 1891 ; Manual of Bacteriology, 1896. Strassmann and Strecker, Zeitschr. f. Medicinalbeamte, 1888. Suringar, Arch. Neerland. 1866 ; Bot. Zeit., 1866. Tappeiner, Fortschr , der Med.. Bd. I. and II. Tataroff, Die Dorpater. Wasserbact., 1891. Tils, Zeitsch. f. Hygiene, Bd. IX., 1890. Tizzoni and Giovannini, Zeigler's Beitrage, Bd. VI., 1889. Tommasoli, Giornale d. Mall. Ven. e. d. Pelle., 1889 ; Monat. t Prakt, Dermat,, Bd. IX. Trambusti and Galeotti, Centralb. f. Bacteriolog., 1892. Tre"cul, Compt. Rend., Ixi. Tricomi, Atti. d. Soc. Ital. de Chir., 1887. Utpadel, Archiv f. Hygiene, Bd. VI. Vandevelde, Studien zur Chemie des Bacillus Sub- tilis; Zeitschr. f. Phys. Chemie, Bd. 8, 1884. Van Ermengem, Bull. Soc. Beige de Microscop., 1888. Van Zieghem, Compt. Rend., Ixxxviii. and Ixxxix., 1879. Vaughan, Amer. Journ. Med. Sci. 1892. Vignal, Archiv de Phys., viii., 1886 ; Le Bacille Mesentericus Vulgatus,- 1889 ; Archiv de Phys., Vol. xxiii. Von Besser, Beitr. z. Path. Anat., Bd. VI. Von Esmarch, Central, f. Bakteriolog., Bd. I. Warington, Journ. of Chemical Soc., 1890. Weibel, Central, f. Bak- teriolog., 1887, 1888, 1893. Weichselbaum, Fortschr. der Med., Bd. V., 1887 ; Ziegler's Beitrage, Bd. IV., 1888 ; Das Osterreich. Samtalswesen, 1889. Weisse, Bull. Phys.-Mathemat. de St. Petersbourg, III., 1845. Weisser, Zeitsch. f. Hygiene, Bd. I., 1886. Welch and Nuttall, Bull. John Hopkins' Hospital, 1892. Welcker, Zeitschr. f. Ration. Med., 1859. Winogradsky, Ann. de I'lnstitut Pasteur, 1890. 1891 ; Bot. Zeitung, 1893. Woodhead and Wood, Compt. Rend., T. cix., 1889. Zelinsky, Proc. Russ. Phys. Chem. Soc., 1893. Zopf, Sitz.-Ber. d. Botan. Ver. d. Prov. Brandenb. : Juni, 1882 ; Die Spaltpilzl. Zimmer- mann, Die Bakt. und Nutz. u. Trinkwasser, 1890. BIBLIOGRAPHY. 665 APPENDICES. ELEMATOZOA. Bardels. Gaz. des Hopit.. 1882. Crookshank, Journ. Eoy. Micr. Soc.. 1886. Cuboni and Marchiafava, Arch. f. Exp. Pathol., Bd. 13, 1880 ; Atti della R. Acad. del Lincei., 1881. Danilewsky, La Parasitologie Comparee du Sang, 1889; Centr. f. Bakt. u. Parasitenk., 1891. Gerhardt, Zeitschr. f. Klin. Med., P,d. 7, 1884. Golgi, Archivio per le Scienze Mediche, 1886. Grass! and Feletti, Centr. f. Bakt. u. Parasitenk., 1891. Klebs and Tommasi-Crudeli, Arch, f . Exp. Pathol.. Bd. 2, 1879. Kotelmaun, Virchow's Arch., Bd. 97, 1884. Laveran, Compt. Kend., 1881 ; Ibid., No. 17, 1882 ; Traite des Fievres Palustres, 1884. Xeoni, Gazetta Medica di Roma. 1*84. Mannaberg, Trans. New Syd. Soc. 1894. Manson, Brit. Med. Journ., 189(5. Marchand, Virch. Archiv, Bd. 88, 1882. Marchiafava and Celli, Atti della R. Academia dei Lincei., 1884 : Fortschr. d. Med., Bd. 3, 1885. Mariotti and Ciarroccbi, Lo Speiimentale, T. 54, 1884. Maurel, Ann. d'Hygiene, 1883. Richard, Compt. Rend., No. 8, 1882. Roszahegyi, Biol. Centralbl., Bd. 2, 1882. Sehlen, Fortschr. d. Med.. Bd. II., 1884. Sternberg, Bull. Nat. Board of Health: Washington, 1881. Tommasi- Crudeli, Arch. f. Exp. Pathol., Bd. 12, 1880; Die Malaria von Rom., 1882 : Conference faite a la 8, Sess. du Congres Intern. Med. a Copenhague, 1884. Torelli, La Malaria in Italia. 1883. Zieb.1, Deutsch. Med. Woch., NT. 48. 1882. COCCIDIA AND CANCER " PARASITES." Albarran, Comp. Rend., 1889. Borrel, Archiv de Med. Exper. et d'Anat. Path., 1890. Bowlby, Brit, Med. Journ., 1891. Cazin, Internat. Congress of Hygiene and Demog., 1891 ; Archiv Gen. de Med., 1892. Darier, Compt. Rend. Soc. de Biol., 1889. Delepine, Brit. Med. Journ., 1891 ; Trans. Internat. Congress of Hygiene, 1891. Foa, Centralb. f. Bakt. u. Parasitenk., 1892. Heidemann, Virchow's Archiv., 1892. Leuckart, Parasites of Man, 1886. Losch, Archiv f. Patholog. Anat., 1875. .Nils Sjobring, Fortsch. der Med.. 1890. Noeggerath, Beitr. z. Struktur u. Entw. des Carcinoms., 1892. Pfeiffer, Die Protozoen als Krankheitserreger, 1890. Podwyssozki and Sawtschenko, Centr. f. Bakt. u. Parasitenk., 1892. Buffer and Walker, Brit. Med. Journ. 1892; Journ. of Comp. Path., Vol. I. Bussell, Brit. Med. Journ., 1890.' Schiitz, Munch. Med. Woch., 1890. Shaltock and Ballance, Proc. Roy. Soc., 1895. Soudakewitch, Ann. de 1'Institut Pasteur, 1892. Wickham, Archiv. de Med. Exper., 1890. SUPPLEMENTARY APPENDIX. EXTRACTS FROM THE FINAL REPORT OF THE ROYAL VACCINATION COMMISSION. THE final Report of the Royal Commission ito inquire into the subject of Vaccination was published on September 18th, 1896.* The author desires to gratefully acknowledge the permission granted him, by the Controller of Her Majesty's* Stationery Office, to make extracts bearing more especially on the history and pathology of protective inoculation and the prevention of small-pox. The reader is recommended by the author to study the whole of the report. History of S The early history of small-pox, like that of many similar diseases, is obscure, is subject to much debate, and, save perhaps on one point, is of antiquarian interest only. The records of the eighteenth century show that the disease was very prevalent in western Europe during the whole of that century. The records of the seventeenth century also show that small-pox was a very common disease during that century : this is especially the case as regards the latter half of the century. The statistics which exist with respect to Geneva, and various scattered statements, further show that small-pox was a well-known disease in the sixteenth century ; but, except for the records which are said to exist of severe epidemics in Iceland taking place as early as 1241, as we go further back the evidence as to the existence of the disease becomes less and less clear, and indeed debate - able, depending as it does largely on the interpretation of incidental * The report may be obtained either directly, or through any bookseller, from Eyre & Spottiswoode, East Harding Street, Fleet Street, B.C., and 32, Abingdon Street, Westminster, S.W. ; or John Menzies & Co., 12, Hanover Street, Edinburgh, and 90, West Nile Street, Glasgow ; or Hodges. Figgis & Co., Limited, 104, Grafton Street, Dublin. f The heading to the extracts from the Report of the Commission are mine. — E. M. C. 067 668 SUPPLEMENTARY APPENDIX. statements in various medical and other writings. There seems, how- ever, to be adequate proof of the prevalence of small- pox in the East, in Asia Minor and other countries, even in the earlier centuries of the Christian era. A view very generally taken teaches that small-pox, introduced from the East, began to be common in western Europe during the fifteenth century, though perhaps existing still earlier; that it increased during the sixteenth and seventeenth centuries, especially the latter ; and that it was very prevalent during the eighteenth century. In dealing with the eighteenth century it must be borne in mind that during the second half of the century the natural conduct of small-pox, as we shall see later on, was modified by the practice of inoculation— that is, by the artificial giving of the disease by the introduction of the virus through a wound in the skin. Our knowledge of the history of small-pox in western Europe during the seventeenth and eighteenth centuries is very largely based on the official records known as the " London Bills of Mortality." Official records bearing on small-pox- are furnished by Geneva, going back as far as the sixteenth century, by Sweden, going back to the year 1749, and by some other places. Data are also furnished, especially for the latter part of the eighteenth century, by parish records in various parts of Great Britain reaching over a variable number of years, as well as by scattered statements in various works. These Bills of Mortality form by far the most complete source of our knowledge of small-pox in England in past times ; but it must be borne in mind that in respect to any contagious disease like small-pox the conditions of London were peculiar. The population was to a marked extent a moving one ; a large number of persons were continually entering London or leaving it, were passing to and from it, from and to the provinces of England and other countries. Of these persons, some, coming from infected districts, brought into London fresh sources of contagion ; others again, coming from districts free from small-pox, and never having had the disease, brought into London fresh material to serve as food for the disease. Further, London presented in an exaggerated degree the two features of a great city which have a great influence on the progress and characters of a contagious disease like small-pox. The crowding both of the dwelling-places and the thoroughfares, as well as the movement continually going on, multiplied the opportunities for the spread of disease, and the accompanying insanitary conditions, as well as the greater inducement to irregular living, tended to increase the severity of the disease when taken, and to heighten the mortality from it. The history of small-pox in London must not be taken as representative of the history of small-pox in England generally. Inoculation of Small-pox. The practice of inoculation for the small-pox— that is, the artificial introduction of the virus into the system by the insertion of fluid from a vanolous pustule into wounds of the skin made for the purpose - began definitely m England towards the end of the first quarter of the REPORT OF THE ROYAL VACCINATION COMMISSION. 669 eighteenth century. Attention was directed to the matter by letters from Timoni of Athens (dated 1713) and Pylarini, published in the 29th volume of the Philosophical Transactions (1716), and especially by a letter from Lady Mary Wortley Montagu in 1717. Though there are indications that in Great Britain and Ireland, as in other countries, some sort of inoculation had occasionally been practised at a much earlier date, the first clearly recorded case in England is that of the daughter of Lady Mary Wortley Montagu (whose son had some time before been inoculated at Constantinople), inoculated by Maitland, in London, in April 1721. Other cases soon followed in England, and about the same time the practice was also introduced in other countries of western Europe, and into the United States of America, namely, at Boston. It was found that the attacks induced by inoculation were as a rule milder, and very much less fatal, than the attacks of the "natural'' disease, the fever and constitutional disturbance being less and of shorter duration, and the eruptive pustules much fewer : the number of these varied, being commonly a dozen or two, somitines only two or three, sometimes a hundred or more. In som i cases there was no eruption at all, the effect being limited to constitutional disturbances and to changes in the wounds of inoculation themselves ; it was maintained that in such cases the disease had really been taken, and immunity against a subsequent attack secured, as in case? of natural small-pox or of inoculated small-pox manifesting itself in an eruption of pustules. In England the practice of inoculation at its introduction, though much lauded and strongly urged by some, was bitterly opposed by others. Moreover, the initial enthusiasm in favour of it soon declined, so that in the years 1730-40 very little inoculation seems to have been practised. About 1740, however, a revival appears to have taken place : in 1746 an Inoculation and Small-pox Hospital was started in London ; and during the whole of the latter half of the eighteenth century the practice may be said to have been very general. It was especially so during the last quarter of the century, the increase being at least largely due to the "improved methods" of inoculation introduced by one Sutton in 1763, and known as " the Suttonian method." Since an inoculated person was infectious, each inoculation was a source of danger to those, not protected by a previous attack, who came into the company of, or even near, the inoculated person during the attack ; and this danger was increased by the fact that the mild character of the inoculated disease permitted, in many cases at least, the patient to move about among his fellows. Moreover, as Haygarth, himself a zealous advocate of inoculation in a systematic regulated manner, points out, the beneficial results of inoculation had robbed the disease of its terrors to so great an extent that the rich and powerful no longer made the efforts which they formerly did to prevent its entrance into, or its spread in, their neighbourhood, and thus favoured its spread among the unprotected poor ; so that inoculation " though eminently useful to the rich appeared to be injurious to the poor." Adding, therefore, together the cases of inoculated small-pox, and the 670 SUPPLEMENTARY APPENDIX. cases of natural small-pox of which the inoculated cases were in one way or other the cause, it seems probable that inoculation did tend to increase the prevalence of small-pox ; but there are no recorded data to show that this really was the case, and this supposed influence may have been counterbalanced by other influences. The evidence as to the influence which inoculation had on the mortality from small-pox is in many respects conflicting. Haygarth, though he admits that in other parts of the kingdom the practice may have saved many lives, was persuaded that in his own part of England and Wales the deaths by the small-pox had been augmented by it ; and he points out that in London, Geneva, and other " towns in different situations and circumstances, the mortality from this distemper has increased since the introduction of inoculation." Several writers in the latter part of the last, and the early part of the present century, held a similar view. Other writers, again, opposed this view. Tradition of the Dairy-folk. There was at the close of the eighteenth century, if not earlier, in districts where cow-pox had appeared, a belief among the dairy-folk that those who had taken the cow-pox never took the small-pox ; and indeed one Jesty, a Dorsetshire farmer, had in 1774, in the case of his wife and sons, purposely introduced the matter of cow-pox into the human subject with the view of protecting from small-pox. Cow-pox. Vaccinia or cow-pox is a disease affecting milch cows, and marked by an eruption on the udder and teats. The disease can be communicated from the cow to man. Dairymen and maids engaged in milking cows affected with cow-pox are apt to have sores of a special kind on their hands or elsewhere, the development of the sores being frequently accompanied by febrile symptoms. There can be no doubt that, in a certain number of cases at all events, such sores are the local manifestations of cow-pox : the virus from the eruption on the cow being introduced into some scratch or other imperfection in the skin of the milker and there producing its local effects, accompanied more or less by general symptoms. Inoculation of Cow-pox. The practice, however, of inoculating with the matter of cow-pox, or vaccination as it was subsequently called, may be considered as dating from the publication of the " Inquiry into the Causes and Effects of the Variolae Vaccine " of Edward Jenner, published in the summer of the year 1798. The practice rapidly spread, and prevailed widely in this country and other parts of western Europe during the first quarter of the present century. It was, beyond all question, so adopted in the genuine belief that it afforded protection against small-pox. In the treatise to which reference has been made Jenner records in the first place a number (19) of cases in which a person who had accidentally taken cow-pox from the cow had never had small-pox, and appeared REPORT OF THE ROYAL VACCINATION COMMISSION. 671 incapable of taking that disease ; the insusceptibility being shown on the one hand by the failure to contract the disease after ample exposure to contagion, such as nursing and attending to or even sleeping with persons suffering from small-pox, and on the other hand by the fact that when the person in question was inoculated with the matter of small-pox in the manner then usual (the matter being tested as to its efficiency on susceptible persons) the inoculation failed to excite small-pox. In the course of the inoculation practice it had been observed that when the operation was performed upon a person who had already had small-pox, either naturally or by inoculation, the wound of inoculation, instead of developing, as it did when the operation was successful in a person who had not had the small-pox, into a vesicle and so into a pustule with the variolous characters (the development being accompanied by febrile symptoms and, save in exceptional cases, by the appearance of a smaller or greater number of variolous pustules on parts of the skin other than the seat of inoculation), presented as a rule nothing more than some slight inflammation, dying away in a few days without any other symptom, or even healed at once without any symptoms at all, local or general ; and in the exceptional cases in which further changes took place in the wound, these were not accompanied or followed by an eruption of pustules or even by the febrile and other general symptoms of small- pox. Accordingly, in cases of small-pox inoculation where it was doubtful whether the disease had been communicated, it had become not an uncommon practice to repeat the operation, in order to judge by the effects produced whether the earlier inoculation had or had not produced the disease ; and the practice, thus originating in connexion with small-pox inoculation, had come to be spoken of as the " variolous test." In his treatise Jenner distinguishes between what he calls true cow-pox and other eruptions which he speaks of as spurious, and which he regarded as not affording protection against small-pox, although he gives no details to show that the cases quoted by him as affording protection were cases of his true cow-pox. He also developed the view that matter derived from horses suffering from the disease known as the grease is capable of giving rise to cow-pox in the cow, and indeed is the real origin of the true disease. It may be added that Jenner also expressed the opinion that the protection thus afforded by cow-pox was permanent in character. Jenner further recorded in the same treatise how he had in 17% inoculated a healthy boy of eight years of age in the arm with cow-pox matter taken from a sore on the hand of a dairymaid who had been infected with the disease by milking cows suffering from cow-pox. He describes the appearances subsequently presented by the wounds, and states that, six weeks afterwards, the results of inoculating the boy with variolous matter were those commonly seen to follow the inoculation of persons who had previously had the cow-pox or the small-pox : that is to say, the " variolous test" showed the boy to be insusceptible to small- pox. Some months afterwards the boy was again inoculated, but no sensible effect was produced on the constitution. Jenner then relates that subsequently, in the spring of 1798, he inoculated a child, and obtained a similar result with matter taken directly from the nipple of a cow infected 672 SUPPLEMENTARY APPENDIX. with cow-pox ; from the pustule on the arm of this child he inoculated another, and from this again several, and from one of these latter a fourth in succession, and then a fifth. To three of these the " variolous test " was applied, and it is stated with the same results. Woodville's Lymph. The experiences of Jenner did not stand alone. His results and views attracted great attention, and in the early part of the year 1799 Woodville and Pearson, who were physicians to the Small-pox Hospital in London, commenced making experiments with vaccine matter with a view to ascertain whether it afforded protection against small-pox. They arrived, like Jenner, at the conclusion that it did. In January 1799 Woodville, having found cow-pox to be present in a " dairy " at Gray's Inn Lane, inoculated seven persons at the Small- pox Hospital with matter from one of the cows at the "dairy," and other persons with matter from sores on a dairymaid employed at the same place who had become infected from the cows. From these cases he inoculated in succession others at the Hospital, eventually to the number of many hundreds, and thus established the stock of what has been spoken of as " Woodville's lymph." Pearson also at the same time occupied himself with the question of inoculation with the cow-pox, writing a pamphlet about it. Woodville and he distributed to many persons in this country and abroad quantities of the lymph from the Hospital ; and this was the beginning of the more general practice of vaccination, for Jenner's stock of lymph, the results of which he had described in his treatise, had come to an end. Although Woodville's "Hospital lymph" appears to have been widely distributed by himself and by Pearson, and thus to have been the source of the lymph used in vaiious places in the early days of vaccination, it was not the only source, even in those days. Pearson also obtained lymph from cow-pox at a dairy in the Marylebone Road, and used this " in certain situations," which may be presumed to include places elsewhere than in the Hospital. He also speaks of having obtained lymph from the cow from a third source. Jenner again, who received and used some of Woodville's Hospital lymph, also obtained lymph from some other sources : for instance, from a cow at a Mr. Clark's farm in Kentish Town. Further, Woodville in 1800 speaks of his having at various times procured the vaccine virus as produced in different cows, which when used at the Hospital produced the same effects as the Gray's Inn Lane lymph. We are not justified in assuming that an account of every new source of lymph was published ; and there may have bsen others, it is impossible to say how many, than those just mentioned. In any case Woodville'.s Hospital lymph was not the only lymph used in those early days ; not only, however, was it largely used (indeed, we have no evidence of so widespread a use of lymph derived from any other source), but the u?e of it marks the definite beginning of the practice of vaccination ; and the history of it demands special notice. REPORT OF THE ROYAL VACCINATION COMMISSION. 673 Of the cases recorded by Woodville in his Reports, the larger number (about three-fifths)ipresented an important, and, as compared with Jenner's cases, a new feature, in that, in addition to the changes taking place at the seat of inoculation and constituting what Woodville called the "cow-pox tumour," which may here be spoken of as the "vaccine vesicle," an eruption over the body of a greater or less number of pustules was observed. These eruptive pustules occurred in the very first cases : of the seven cases inoculated from the cow, four, and of the five inoculated from the dairymaid, four, had such pustules ; and their appearance is recorded again and again in the series, down to the case which appears last but one in the tabular statement forming part of the Reports. Moreover, an eruption of pustules is described in certain of the cases of which accounts were published at about the same time by Pearson and many others. In some of these cases the lymph used was supplied from the Small-pox Hospital by Woodville or Pearson. It must be admitted that these pustules were pustules of small-pox,, and that, therefore, Woodville's cases, which did so much to establish the practice of vaccination, were not cases simply of cow-pox but of cow-pox mixed, so to speak, with small- pox. It has indeed been maintained that Woodville's cases were not cases of cow-pox at all — that small-pox was inadvertently introduced into the very first cases ; that the history of the whole series is the history of a series of small-pox cases putting on special characters, and that therefore the lymph used and distributed by Woodville and Pearson was in reality not cow-pox lymph but small-pox lymph. A review of all the evidence available leads to no other conclusion than that, however much in Woodville's, Pearson's and other cases cow-pox was mixed up with small-pox, the lymph used and distributed by Woodville and Pearson and called by them cow-pox lymph (excluding of course all the cases, of which there were not a few, in which matter was taken not from the local ;; cow-pox tumour" at the seat of inoculation, but from one of the eruptive pustules) was veritable cow-pox lymph having the true characters of cow-pox lymph only. It of course follows that the cases, both in Woodville's practice and in that of others, in which the inoculation of cow-pox matter wa& accompanied by an eruption of pustules, due to small-pox being present as well as cow-pox, when appealed to as showing immunity against small-pox (by the test either of exposure to contagion or of inoculation), furnished false evidence as to that immunity being due to cow-pox ;. it might have been due to the accompanying small-pox. So far then as the adoption of vaccination was assisted by cases of this description, it may be held to have rested' on erroneous data. The Decline of Small-pox. One effect of the introduction of vaccination was a very great decrease in the practice of inoculation, which had become very prevalent during the later part of the previous century. And the view has been put forward. 43 674 SUPPLEMENTARY APPENDIX. that, the prevalence of inoculation having greatly increased the amount of small-pox, the diminution of small-pox in question was the result of the decrease of inoculation. The question how far the behaviour of small-pox in the eighteenth century and earlier was influenced by sanitary conditions, is one rendered difficult by the lack of exact information. We may distinguish between overcrowding as one insanitary condition and all other insanitary condi- tions, such as lack of cleanliness and the like. A priori we should ex- pect that a dense population, especially one of great internal movement, and one in continual interchange with surrounding populations, by offering greater facilities for the conveyance of contagion, would lead to a greater amount of small-pox. London was a conspicuous instance of the above, and the apparent greater prevalence of small-pox in London than in the provinces may be attributed to these causes : but it would appear that the increase was felt — as indeed would, a priori, seem pro- bable— rather in the constant presence of small-pox to a considerable amount at all times than in the mortality of the epidemics when these occurred. And the same seems also to be shown to a less extent in other large cities, such as Liverpool. But in this matter of dense and moving populations the eighteenth century did not differ markedly from the early part of the nineteenth. We might a priori expect the other acknowledged imperfect sanitary conditions of the eighteenth century to increase the fatality of, and so to a corresponding extent the mortality from, small-pox ; but there is no exact evidence to confirm this supposi- tion. If on the contrary we recognise that in the course of the eighteenth century the general mortality, the relative number of deaths from all causes, went on decreasing, and attribute, as has been done, this decrease to improved sanitary conditions, no like decrease of small-pox took place. Again, the places which were deemed the most salubrious appear to have been visited by epidemics of small-pox as severe as those which fell on unhealthy places. Thus the epidemic in Chester in 1774 was undoubtedly a severe one, and yet Haygarth writes, " The healthiness of Chester," as shown by statistics, " must appear so very extraordinary as to be almost incredible." And in general both the incidence of, and mortality from, small-pox seem to have been far less affected by sanitary conditions than might a priori have been expected. It may be urged against the view that the decline of small-pox was due to improved sanitary conditions, in the first place, that, admitting the introduction of sanitary improvements, no evidence is forthcoming to show that during the first quarter of the nineteenth century these improvements differentiated that quarter from the last quarter, or half, of the preceding century in any way at all comparable to the extent of the differentiation in respect to small- pox. In the second place, admitting a priori that crowded dwellings tend to increase the liability to contagion, and so the prevalence of the disease, while other insanitary conditions tend in addition to increase the fatality among those attacked, that insanitary conditions as a whole must tend to increase the mortality from small-pox,— no evidence is forthcoming which distinctly shows that the dependence of the prevalence of, or the mortality from, REPORT OF THE ROYAL VACCINATION COMMISSION. 675 small-pox, on the lack of sanitary conditions, was a feature of the history of small-pox during the eighteenth century. Upon the whole, then, we think that the marked decline of small-pox mortality in the first quarter of the present century affords substantial evidence in favour of the protective influence of vaccination. Age Incidence of Small-pox. A study of the age incidence of small-pox mortality is very instructive. In connexion with this point it is necessary to bear in mind that experi- ence has led to the conclusion that, whatever be the protective effect of vaccination, it is not absolutely permanent ; the most convinced advocates of the practice admit that after the lapse of nine or ten years from the date of the operation its protective effect against an attack of small-pox rapidly diminishes, and that it is only during this period that its power in that respect is very great ; though it is maintained that, so far as regards its power to modify the character of the disease and render it less fatal, its effect remains in full force for a longer period, and never altogether ceases. The experience upon which this view is founded is derived almost exclusively from the case of infantile vaccination. It has been supposed by some that the transitory character of the protection results from changes connected with the growth from infancy to adult years. Whether this be so or not, we have no means of determining. No doubt, when Jenner drew the attention of the public to the value of vaccination, he believed that a single successful inoculation of vaccine matter secured absolute immunity for the future from an attack of small- pox. It is certain that in this he was mistaken. It may well be doubted whether the anticipation was a reasonable one. No such immunity is secured by an attack of small-pox, though there are few who would maintain the proposition that it is without protective influence against another attack. A priori there would seem to be no sound ground for expecting that vaccinia would afford more potent protection than small- pox itself. The extent of the protection afforded (assuming that there is some protective influence) could only be determined by experience. It soon became apparent that Jenner had, in the first instance, overrated the effect of vaccination. That he should thus have overestimated it is not to be wondered at, when the tendency to be unduly sanguine, which besets the discoverer of any new prophylactic, and, indeed, every discoverer, is borne in mind. We think, taking it all together, that the evidence bearing upon the question whether the vaccinated are less liable to be attacked by small-pox than the unvaccinated, points to two conclusions : first, that there is, taking all ages together, less liability to attack among the vaccinated than among the unvaccinated ; and next, that the advantage in this respect enjoyed by vaccinated children under ten years of age is greatly in excess of that enjoyed at a more advanced period of life. In considering whether vaccination has been the principal cause of the decline, we must inquire whether the other causes suggested by those who deny the efficacy of vaccination will satisfactorily account for it. 676 SUPPLEMENTARY APPENDIX. Effect of Sanitation. It is said that the decline has, in the main, been due to changes in the general conditions of life in the different parts of the United Kingdom, apart from the spread of the practice of vaccination,— amongst other things, to improvement of sanitary conditions. It is beyond doubt that an infectious disease like small-pox is, other things being equal, more likely to spread in towns than in country districts, and more likely to spread in crowded town districts than in others not so densely populated ; so that we should expect a lessened proportion of overcrowded dwellings, by diminishing the opportunities for contagion, to check the prevalence of the disease and consequently to render its mortality less. Effect of Isolation. It has been maintained that the decline in small-pox mortality is largely due to more frequent and systematic attempts to isolate those suffering from small-pox. We think an answer to this contention is to be found in the fact that it is only in quite recent years that there has been any systematic practice of isolating small-pox patients, and that it has been confined even then to a very limited number of localities. The fact to which we are about to call attention in greater detail than hitherto, that the decline in the deaths from small-pox is -found almost exclusively among those of tender years, appears also to militate against the contention. The risk of contagion is not confined to children. Adults also are subject to it. If a better system of isolation had been a main cause of the reduced mortality, we should have expected to see it operate in the case of adults as well as of children. At the same time we are far from thinking, as will appear when we come to deal with that subject, that the efforts at isolation which have characterised recent years have been without a beneficial effect on small-pox mortality. Sanitary Legislation. We have already pointed out that on a priori grounds it is reasonable to think that improved sanitary conditions would tend to diminish the fatality of, and so to a corresponding extent the mortality from, small-pox. And there can be no doubt that the period with which we are dealing has been characterised by an improvement of this description. There has been better drainage, a supply of purer water, and in other respects more wholesome conditions have prevailed. It may be useful at this point to furnish a brief summary of the principal Sanitary Acts which have been passed relating to the different parts of the United Kingdom. In 1848 was passed the first great and comprehensive measure which may be called the groundwork of our sanitary legislation as regards England. The Public Health Act of 1848 was, however, principally designed for towns and populous places in England and Wales, not including the Metropolis, which was dealt with in Acts passed in the same year. The powers of local government supplied by the Act were generally an extension of those before given by sundry local Acts to REPORT OF THE ROYAL VACCINATION COMMISSION. 677 -Commissioners of Sewers in the Metropolis, and to authorities in a few large towns. Many provisions corresponding to sections in the Towns Improvement Clauses Act of 1847 are found in the Public Health Act, and communities were thus enabled to obtain by a simple process powers which they, could not previously obtain except by a local Act incorporating sections of the Towns Improvement Clauses Act. In 1848 was also passed the Nuisances Removal and Diseases Preven- tion Act of that year, in substitution for a similar Act of 1846 which was about to expire ; and in 1849 this Act of 1848 was amended. The provisions of all these three Acts extended to England, Scotland and Ireland. In 1855 a comprehensive Nuisance Removal Act was, as regards England, substituted for the Acts passed in 1848 and 1849 ; and in the following year there was similar legislation for Scotland. In 1860 the English Act was amended ; and in 1866, by the Sanitary Act of that year (to which we shall again refer), the provisions of the English Acts of 1855 and 1860, as then amended, were applied to Ireland. In 1855, by the Metropolitan Local Management Act of that year, provision was made for the appointment of a medical officer of health and an inspector of nuisances by every vestry and district board in the Metropolis. This provision did not extend to the City of London, where, in 1848, a medical officer of health had been appointed under power given by a local Act. In 1858, the Local Government Act of that year, to be construed with the Public Health Act of 1848 as one Act, was passed, and took effect in all places where that Act was in force at the time of its passing ; and, as regards England, these two Acts together constituted until 1872 the principal sanitary legislation on the statute-book. There followed, however, within the next ten years many public Acts having sanitary objects, some applying to all, and some to particular, parts of the United Kingdom, besides numerous other Acts of local application. We need only now specially refer to one of these public statutes — the Sanitary Act of 1866, which was probably the most important, and applied, in part at least, to England, Scotland and Ireland. This Act, amongst other things, extended the powers of local authorities for the disposal of sewage, and, in amending the English Nuisances Removal Acts of 1855 and 1860, added to the definitions of nuisances, especially as regards crowded houses and workshops, and to the duties and powers of local authorities for their abatement, especially in the way of providing means for disinfection and places for the reception of dead bodies. In 1867 the Public Health (Scotland) Act was passed — a comprehensive measure which consolidated into one Act, with certain amendments, the whole statute law relating to the public health in Scotland. In 1872 a complete distribution of England into sanitary districts took place, and some further amendments were made in the sanitary laws. In 1875 these laws were consolidated in the Act of that year. In 1891 a Sanitary Act was passed relating to the Metropolis. In 1874 an Act was passed for Ireland, containing substantially the same provisions as those which had been enacted in the case of England in 1872. 678 SUPPLEMENTARY APPENDIX. Value of Vaccination. We have not disregarded the arguments adduced for the purpose of showing that a belief in vaccination is unsupported by a just view of the facts. We have endeavoured to give full weight to them. Having done so, it has appeared to us impossible to resist the conclusion that vaccination has a protective effect in relation to small-pox. We think :— 1. That it diminishes the liability to be attacked by the disease. 2. That it modifies the character of the disease, and renders it («) less fatal, and (6) of a milder or less severe type. 3. That the protection it affords against attacks of the disease is greatest during the years immediately succeeding the operation of vaccination. It is impossible to fix with precision the length of this period of highest protection. Though not in all cases the same, if a period is to be fixed, it might, we think, fairly be said to cover in general a period of nine or ten years. 4. That after the lapse of the period of highest protective potency, the efficacy of vaccination to protect against attack rapidly diminishes, but that it is still considerable in the next quin- quennium, and possibly never altogether ceases. 5. That its power to modify the character of the disease is also greatest in the period in which its power to protect from attacks is greatest ; but that its power thus to modify the disease does not diminish as rapidly as its protective influence against attacks, and its efficacy during the later periods of life to modify the disease is still very considerable. 6. That re -vaccination restores the protection which lapse of time has diminished; but the evidence shows that this protection again diminishes, and that, to ensure the highest degree of protection which vaccination can give, the operation should be at intervals repeated. 7. That the beneficial effects of vaccination are most experienced by those in whose case it has been most thorough. We think it may fairly be concluded that where the vaccine matter is inserted in three or four places, it is more effectual than when introduced into one or two places only — and that if the vaccination marks are of an area of half a square inch, they indicate a better state of protection than if their area be at all considerably below this. Question of Specific Protection or of Antagonism. When an attack of disease secures immunity or protection against another attack of disease, the two attacks are, as a rule, attacks of the same disease. Some pathologists have, it is true, of late years been led to suppose that one disease may confer some degree of immunity or protec- tion against another different disease ; but instances of this are few, andr moreover, cannot be regarded as thoroughly established. The ordinary instances of immunity are so clearly those in which the attack, natural or REPORT OF THE ROYAL VACCINATION COMMISSION. 679 artificial, which confers the immunity is of the same disease as that towards which immunity is conferred, that identity of disease has been considered as essential to the conferring of immunity. And it has been argued that it is a priori improbable that cow-pox should confer immunity from small- pox, seeing that the two are different diseases. Such a purely theoretical argument can have little weight against positive evidence of vaccination having actually conferred immunity. If this be definitely proved to be the fact, proof is thereby at the same time afforded that the theory is unsound, either because a particular disease may confer immunity against a different disease, or because small-pox and cow-pox are not different diseases. For the practical object with which alone we are concerned, it is not material that we should reach any conclusion upon the question what is the real source of error in the theory alluded to, supposing it to be erroneous ? We shall content ourselves, therefore, with a very brief notice of the subject. It appears to us that we may dismiss for practical purposes the theoretical questions which were discussed before us so fully. If the fact be established that the introduction of vaccine matter and the consequent vaccinia produce some effect upon the human body which renders it less susceptible to small-pox, or which modifies that disease when the small-pox virus enters the system, it will not be a strange or unwonted experience that we should be unable to explain how this comes about. Science has not yet succeeded in freeing therapeutics or kindred subjects from obscurity, or in solving all the problems which they present. The precise modus operand}, by which a previous attack of a disease furnishes security against a subsequent attack by the same disease has not yet been elucidated. There can be no cause for astonishment, then, if we are unable to trace the steps by which vaccination exerts a protective influence, supposing the fact that it does so be established, nor is it essential that we should succeed in tracing them. Our inability to accomplish this does not seem to us to be the slightest reason for regarding with doubt the conclusions to which the facts lead us. Professor Crookshank, than whom no one has more strongly insisted on the theoretical arguments against the protective influence of vaccination in relation to small-pox, gives it as his opinion that vaccination creates a transient antagonism to that disease. We understand his view to be that an attack of disease can only afford protection against the same disease, and that small-pox and cow-pox are not the same but different diseases. We gather, however, that, in his opinion, so long as the state of antagonism lasts, the person in whose system it exists is less likely to suffer from small-pox than he would be if the state of antagonism were wanting. This seems to us to amount in effect to the same thing as saying that during that period vaccination has conferred some protection. Whether the effect be to create antagonism or to confer protection, and whatever difference there be between the mod tin ojtertntdi in the one case and in the other, we know equally little about it. If a condition of transient antagonism to small-pox is induced by vaccination, theoretical considera- tions will not afford a guide of the slightest value to the conclusions how long this transient antagonism will last, or how soon it will pass away. SUPPLEMENTARY APPENDIX. Experience, and experience alone, can answer that question. A priori we -do not see that there is any better reason for supposing that it would last for two or three years than that its duration would extend to ten years. Cow-pox and Small-pox not Convertible. Jenner himself, in his first paper, advanced the view that the cow-pox and small-pox were identical with each other ; and since his time numerous observers have attempted to prove the identity of the two diseases experimentally — namely, by giving rise to cow-pox in the cow through the inoculation of small-pox matter, or by the introduction of contagion in other ways. It may at once be stated that while cow-pox is readily transferred from the cow to man and back again from man to the cow, the disease in man being identical with that in the cow, small- pox cannot be transferred from man to the cow so as to give rise to a disease in the latter identical in its features with the small-pox of man. ^Nor can cow-pox be so transferred to man as to give rise in him to small-pox. The two diseases are not in this sense convertible. Small-pox Vaccine. In most cases the attempt to transfer small-pox from man to the •cow has had simply a negative result ; no obvious effect of any kind has been observed. This has been the case in the attempts to introduce the contagion through absorption by the respiratory or digestive organs, and in most of the attempts to introduce the contagion by inoculation. In certain instances these latter attempts have produced results which may be briefly described in three categories. (We may pass over the isolated experience of Thiele, who in 1838 asserted that by keeping small-pox matter sealed between glass plates for ten days before using it, and by diluting it with milk when using it for inoculation, the matter thus treated through ten removes through the human body — the cow not intervening at all — was converted into something which gave results identical with those of ordinary vaccine matter. We are not aware of any attempt to corroborate this experiment.) The first category includes the experiments in which the inoculation of small-pox matter into the udder, or adjoining parts, of the cow gave rise at or near the seat of inoculation to a vesicle, either identical in visible characters with the ordinary vaccine vesicle produced by inocula- tion with the matter of cow-pox, or to a vesicle the features of which, while not corresponding wholly with those of a perfect vaccine vesicle, so closely resembled them as to justify the vesicle being called a vaccine vesicle. Further, the matter from a vesicle which at the first inoculation had not the characters of a perfect vaccine vesicle, when carried through a second or third remove in the cow, fully acquired those characters, and when transferred to man gave results indistinguishable from the ordinary vaccine vesicle. Indeed, lymph of such an origin has come into general use for vaccination purposes. Of the experiments, the best known or most quoted are those of Thiele (1838), Ceely (1840), Badcock (between 1840 and 1860), Voigt (1881), Haccius and Eternod (1890), King (1891), REPORT OF THE ROYAL VACCINATION COMMISSION. 681 Simpson (1892), and Hinie (1892) ; but there are several others. The details of the experiment are very scanty in the cases of Thiele and Badcock, but more full in the others, especially, perhaps, those of Ceely and Haccius. In the second category may be placed the experiments of Klein and Copeman. Klein, who had in 1879 obtained in 31 trials what then appeared simply negative results, found in a renewed research in 1892 that the result of the first inoculation in the cow of small-pox matter was not a distinct vesicle but merely a thickening and redness of the wound. Lymph pressed from the thickened wound produced, when inoculated into a second cow, a like result, but rather more marked ; the thickening and reddening still further increased with a third and a fourth cow. Lymph squeezed from the wounds of the fourth cow produced in a child typical vaccine, and crusts from the child produced typical vaccine in a cow. Copeman obtained somewhat similar results ; the appearances increasing in three removes and approaching those of typical vaccine, but not reaching them. The third category consists of -ttye results obtained in an elaborate inquiry conducted by a Commission of the Society of Medical Sciences at Lyons, with Chauveau at its head. Those results, reported in 1 865 , were briefly as follows : — Inoculation of the cow with small-pox matter in any one of the 30 animals used did not give rise to a vaccine vesicle. Nevertheless a definite result was obtained ; in the form, however, not of a vesicle, but of a thickening and inflammation of the wound ; when a puncture was employed this became a papule. Lymph squeezed from such a papule and inserted into a second animal gave rise to a like papule : and this, again, might be used for a third animal, but often failed ; and the effect could in no case be carried on through more than three or four removes. When the inoculation was repeated on an animal in which a previous inoculation had produced such a papule, no distinct papule was formed ; and, moreover, lymph squeezed from the seat of inoculation produced no effect at all when used for the subsequent inoculation of another animal. This shows that the development of the papule was the result of the specific action of the virus. The same is shown by the fact that no such papule was produced when the small- pox matter was inserted into an animal which had previously had cow-pox naturally or artificially, as well as by the fact that when an attempt was made to vaccinate, with vaccine matter of proved efficacy, an animal on which a papule had been so developed by inoculation with small-pox, the vaccination failed, though the animal had never had natural cow-pox or had never been vaccinated. The specific nature of the lymph of the papule is further shown by the fact that such lymph when used on the human subject gave rise to veri- table small-pox. It has been urged that in this case the virus producing the effect was simply the old virus used in the inoculation, producing the papule and still clinging to the wound. This is disproved by the experience that lymph from a ))ul<' <>f the second re more also gave rise in the human subject to veritable small-pox. Thus Chauveau and his Commission found that small-pox implanted 682 SUPPLEMENTARY APPENDIX. in the cow gave rise to a specific effect which was not cow-pox but was of the nature of small-pox, though its manifestations in the cow were different from those of small-pox in man. They also obtained similar results in attempting to transfer small-pox to the horse. It must be admitted that the results finally obtained in some of the successful cases were indistinguishable from those of vaccination ; the characters of the local vesicle, the absence of eruptive pustules and of contagiousness, show that the lymph thus apparently originating from small-pox in the hands of Ceely, Badcock, and others, was vaccine lymph. It has been urged that a vaccine vesicle making its appearance in the wound of inoculation with small-pox was due to the accidental intro- duction of cow-pox matter into the wound ; the small-pox matter in the wound produced no effect, and the cow-pox matter its usual effect. Several considerations support this view. The cow is peculiarly suscep- tible to cow-pox. In some cases (Ceely, Voigt), the animal was vaccinated as well as inoculated with small-pox : thus, in Ceely's first case, the animal was inoculated with small-pox on one side of the body, and a few days after vaccinated on the other side. In many cases the experiments were conducted in an animal vaccine establishment, the stalls, the operating tables, and the assistants being those used or engaged in vaccination. It is true that in some cases at least special precautions, sterilisation of instruments and the like, were taken to avoid the accidental introduction of cow-pox ; but in observations of this kind the difficulties of avoiding all such sources of error are notorious. Still the successful cases are now so numerous that it is difficult to resist the conclusion that the same accident could not have occurred in all, and that a transformation of small-pox into cow-pox — that is to say, into the artificially inoculated cow-pox which we call vaccine * — really took place. Accepting this view provisionally, it may be remarked that in most cases the transformation was sudden and complete ; the small-pox virus, under the influence of the tissues of the cow, became immediately converted into vaccine virus, and this produced a typical vaccine vesicle. In some cases (ex. gr., that of Hime) the transformed virus produced its effect not in the wound of inoculation, or not chiefly so, but at some little distance from it. In some cases the characters of the vesicle first formed, though sufficiently distinct to justify the vesicle being called a vaccine vesicle, were not those of a perfect vaccine vesicle, but the lymph from such a vesicle, at least after one or two removes, gave rise to most typical vaccine vesicles. In Klein's experiments the transformation was gradual. In his fourth cow, the virus was as yet not typical vaccine, since it did not produce a typical vesicle ; yet it was so far already vaccine that, transferred to the child, it produced typical vaccine (unless we suppose some accidental introduction of vaccine to have taken place). That the vesicle on the child was vaccine, and not small-pox unaccompanied by eruptive pustules, was shown not only by its characters but also by the fact that lymph from it produced typical vaccine in the cow. In "Chauveau's experiments no transformation at all took place. * The italics are mine. — E.M.C. REPORT OF THE ROYAL VACCINATION COMMISSION. 683 As has been urged in another place, there are no adequate reasons- leading us to believe that in the human subject the small-pox virus and the cow-pox virus can so act on each other as to form a hybrid disease. But this does not preclude the view that, accepting the conclusion that the body of the cow has the power to convert small-pox into vaccine, the virus may exist for a while in a phase in which, while' ceasing to be typical small-pox, it has not yet fully acquired the characters of vaccine, and we may regard Klein's results as illustrating this. In some of the experiments — for instance, those of Ceely and Voigt — the results obtained with the lymph of the vesicle produced by the inoculation of small-pox give rise to the suspicion that the lymph had small-pox qualities, as seenr for example, in the case of Ceely's assistant, Taylor ; but the facts cannot be said to be more than suspicious — they are not decisive. Moreover, admitting that the vesicle itself in such cases was the result of the trans- formed virus, some not transformed old virus might still remain dormant in the wound, and might be present in the lymph of the vesicle, mixed with the transformed and generating virus ; this old virus might have happened to be in excess on the point of the lancet which wounded Taylor. Small-pox Vaccine — Cow-pox Vaccine — Horsebox Vaccine — Cattle- plague Vaccine — Sheep-pox Vaccine. Taking all the various facts into consideration, we seem led to the provisional conclusion that under certain conditions the tissues of the cow are able to transform small-pox into vaccine, that these conditions may be such as to lead to the transformation being sudden and complete, that under certain other conditions the transformation may be gradual and incomplete, and that under certain other conditions (and these seem most commonly to obtain) the transformation into vaccine does not take place at all. But what the above conditions are has not as yet been clearly made out. It has been suggested that one condition favourable to the transformation is extreme youth of the subject : to effect the change the animal used should be a calf of not more than three or four months old : but this is not definitely proved. Until these favourable conditions have been clearly recognised, so that, the conditions being fulfilled, the transformation will always be secured, the conclusion cannot be regarded as indisputable. Moreover, it must be borne in mind that effects more or less closely resembling a vaccine vesicle have been at various times obtained by various observers through inoculating man or the cow or another animal with material other than that obtained from the pustules of the small-pox of man. Much discussion has taken place concerning the " grease " of the horse, which Jenner believed to be the origin of the cow-pox of the cow. Without entering into any discussion of the matter, it may be said that investiga- tion has shown that horses do suffer from a malady which, transferred to the cow, gives rise to a vaccine identical apparently with that produced by the inoculation of the natural cow-pox. Hence this malady is spoken of as the " horse-pox," and some cases at least of so-called " grease " appear to be cases of this horse-pox. But it is at least not proved that 684 SUPPLEMENTARY APPENDIX. all the cases of " grease " which by inoculation were found to give rise to vaccine vesicles in man were cases of true horse-pox. And this at least must be said, that no investigations as complete and varied as those which have been carried out with regard to the development of vaccine vesicles through the inoculation of small-pox matter, have been carried out with regard to the alleged development of vaccine vesicles by tho inoculation of other material, such as the matter from the eruptions of the sheep-pox, the cattle plague, and the like. Nor have there been like extended in- quiries as to the production of vesicles resembling those of vaccine by the inoculation of small-pox matter into animals other than the cow or the horse ; such results as have been obtained by observers are conflicting. 'There is still room for much inquiry ; meanwhile it may be said that, in any case, the evidence in favour of a possible transformation of small-pox into vaccine is sufficiently strong to remove the force of the theoretical objection to the power of vaccination to secure immunity towards small- pox, on the ground that the two diseases are absolutely distinct. / Risks of Vaccination. It must not be forgotten that the introduction into the system of even a mild virus, however carefully performed, is necessarily attended by the production of local inflammation and of febrile illness. If these results did not in some measure follow, the practice would probably fail in its protective influence. As a rule, the inflammation and illness are of a trifling character ; in exceptional cases, however, they may exhibit more severity, and, as certain facts submitted to us in evidence have shown, there are cases, though these are rare, where a general eruption may follow vaccination. In order to determine how far the risk of erysipelas is a necessary incident of vaccination, what is the extent of that risk, and how it may best be avoided, it is necessary to consider the various circumstances which may occasion erysipelas and allied diseases in the case of vaccinated children. It is established that lymph contains organisms, and may contain those which under certain circumstances would be productive of erysipelas. It is therefore possible that some contagious material (the specific virus of erysipelas, for instance,) may be conveyed at the time of vaccination, owing either to its presence in the lymph employed, or to its being conveyed by the vaccinator himself, or by those with whom the child comes in contact at ,the time of vaccination. We believe that the cases in which the virus is conveyed at the time of vaccination are rare. It has, however, in some instances been clearly established, the immediate occurrence of erysipelas in several co-vaccinees making it practically certain that some virus was conveyed at the time of the operation. In some instances where this has been the case, and there is every reason for believing that the contagion was conveyed through the medium of the lymph, it is nevertheless in evidence that the vaccinifer did not display anything more than a slightly inflamed arm. The scrupulous avoidance of inflamed arms in vaccinifers will do much to reduce the risk of •conveying erysipelas in the act of vaccination (a risk which, as we have REPORT OF THE ROYAL VACCINATION COMMISSION. 685 seen, has been proved to be a very slight one), but it is possible it would not wholly remove it. We have dwelt upon features presented by the cases of erysipelas and various forms of septic disease which have followed vaccination, because they suggest precautions which may be adopted to lessen, if not to prevent, such evils in the future. If, for example, vaccination were performed at the patient's home instead of at a public vaccination place, the chance of disease being contracted at the time of vaccination would be to some extent diminished ; and the same may be said of the inspection of the vaccinated person which takes place eight days after the operation. On these points we shall have some remarks and recommendations to make at a later stage of our report. A study of the cases which have been made the subject of careful examination and report points to the conclusion that an exercise of greater care would largely diminish the risk, already small, of erysipelas- contagion and blood-poisoning. Although it may be confidently hoped that by additional care on the part both of vaccinators and parents, the number of inflamed arms and of cases of erysipelas may be reduced to very few, yet it is not to be expected that such occurrences will be wholly prevented. A vaccination wound is, like one from anv other cause, so long as it exists, a source of some risk. The use of calf-lymph, though it may be supposed to be more free from the risk of conveying erysipelas, does not appear to prevent inflamed arms. Some witnesses have indeed supposed that it is attended with more risk of inflammation than the employment of that taken from the human subject. This opinion has not, however, been corroborated by some of those of widest experience. Nothing has produced so deep an impression hostile to vaccination as the apprehension that syphilis may be communicated by it. It was at one time doubted whether syphilis could result, and it was even confi- dently asserted that it could not. The fact that this was possible had been fully established, and was generally acknowledged by the medical profession, before we commenced our inquiries. The very close resemblance in certain very rare cases of the results of vaccination, whether with calf-lymph or humanised lymph, to those attributed to syphilis (a resemblance so close that it has caused in a few cases a difference of opinion whether the disease was syphilis or vaccinia) has led to the expression by Dr. Creighton of the opinion that there is some essential relationship between the two diseases. This, however, is a point of speculative, almost it might be said of transcendental pathology, upon which for practical purposes it is useless to enter. It must be sufficient to remark that, whatever may be the relationship alluded to, it exists, if it exists at all, equally between small-pox and syphilis as between vaccination and syphilis. For all practical purposes variola and vaccinia are both wholly distinct from syphilis, and their differences are, with the rarest exceptions, easily recognised. They are alike in being attended by affections of the skin and mucous membranes, and exceptionally by disease of the bones, eyes, and other parts ; but in all 686 SUPPLEMENTARY APPENDIX. these it is a question of resemblance and not of identity with which we have to deal. Only a few items of the evidence produced before us appear to require special notice : among these, the most prominent is what has been known as the " Leeds case," upon which we have heard the evidence of Mr. "Ward, Mr. Littlewood and Dr. Barrs. The witnesses named regarded it as a case of syphilis, conveyed by vaccination, but all of them admitted that the course of events was most unusual. We have carefully investigated this case, and notwithstanding the opinion formed by the witnesses, there appears good reason to doubt whether it was one of syphilis. The case was made the subject of careful inquiry by Dr. Barlow on our behalf, who shared the doubt we have expressed. The view taken by the medical inspector of the Local Government Board who in the first instance in- vestigated the case was that it was a case of hereditary syphilis. It seems certain, however, that the parents of the child whose death was in question were not in any way affected with syphilis. The vaccinifer also appeared to be free from any taint of that disease, and its family history confirmed this view. The co-vaccinees from the same lymph also exhibited no trace of syphilis. These facts of themselves make out a strong case against that having been the nature of the disease. Coupled with the fact that it could not have been communicated by the vaccinator himself, they seem to render it practically impossible that syphilis was the cause of death. If the symptoms exhibited had in all respects corresponded with those which are known to characterise syphilis, the proper inference might have teen that there was some error in ascertaining the facts of the case. But it is beyond question that the course of events was very different in some respects from that experienced in undoubted cases of syphilis, and we think the true conclusion is that it was not a case of that disease. It may probably be classed with a few others as examples of gangrene and blood- poisoning, the direct result of vaccination, which are not to be explained by supposing the introduction of any syphilitic or other poison. Fortu- nately, such cases are extremely rare — so much so that the witnesses concerned knew of no case precisely parallel. The evidence offered to us would lead to the belief that, whilst with ordinary care the risk of communication of syphilis in the practice of arm- to-arm vaccination can for the most part be avoided, no degree of caution can confer an absolute security. The rejection as vaccinifers of young infants, say below four months of age (in whom congenital syphilis may be as yet undeclared), and of adults (in whom the disease may possibly have been recently acquired) are precautions which would probably shut out almost the whole of the risk. The outbreaks of syphilis in connection with vaccination which have been mentioned to the Commission (all of which had been previously published) have occurred chiefly in arm-to-arm vaccination amongst soldiers, or from the use as vaccinifers of young infants the offspring of parents whose history was not known to the vaccinator. It must, however, be admitted that neither the examination of the vaccinifer if taken alone, and without a knowledge also of the parents, nor the most scrupulous avoidance of any visible admixture of blood with the lymph, are in themselves, however valuable, sufficient REPORT OF THE ROYAL VACCINATION COMMISSION. 687 absolutely to exclude risk. The evidence given by Dr. Husband, of the Vaccine Institution of Edinburgh, established the fact that all lymph, however pellucid, does really contain blood cells. Absolute freedom from risk of syphilis can be had only when calf-lymph is used ; though where the antecedents of the vaccinifer are fully ascertained, and due care is used, the risk may for practical purposes be regarded as absent. It is obvious that the employment of calf -lymph only would wholly exclude the risks as regards both syphilis and leprosy. Respecting the latter disease, however, there appears to be reason to doubt whether any risk exists, and at any rate it .does not concern the British population. Even in leprosy districts the employment of English human lymph would be, so far as leprosy is concerned, as safe as that from the calf. There can be no doubt that vaccination ought to be postponed when erysipelas, scarlet fever, measles, or chicken-pox are prevalent in the neighbourhood of the child's residence, or, if the child is not to be vaccinated at home, either there or near the place of vaccination. Here again there would be a gain if the home was more often the place of vaccination. It would, in our opinion, be an advantage if the postponement of vaccination were expressly permitted, not only on account of the state of the child, but of 'its surroundings and any other conditions rendering the operation at the time undesirable. If more discretion in this respect were possessed and exercised, we think untoward results would become even rarer than they are. We are quite alive to the objections which may be urged against a prolongation of the period within which vaccination must be performed. It will naturally be said that a number of children, who otherwise would be protected against small-pox, would be left without that protection, and would thus be liable to suffer from the disease themselves, and be a source of danger to others. It must be remembered, however, that so long as children cannot walk, the risk of their contracting contagion is less than if they were able to move freery about and mix with other people, and that, for the same reason, the risk of their communicating contagion to others is less. We cannot trace in the statistics relating to Scotland any grounds for believing that the later compulsory vaccination age which prevails in that country as compared with England has affected, to any substantial extent, the general small-pox mortality of Scotland, though no doubt it may have led to some deaths among children under six months of age which otherwise would not have taken place. We have already shown how satisfactory a position Germany has occupied in relation to small-pox since the year 1874. The age of com- pulsion in that country is the end of the next calendar year after birth. It is true that re- vaccination has been there made compulsory as well as primary vaccination ; but we think the experience of Germany is not without its bearing on the question we are now considering. Wherever the line is drawn, whether at three months or six months, it will always leave a class of unvaccinated persons. The age to be fixed is a question of policy into which many considerations must enter. If an extension of the age within which vaccination was required rendered its untoward 688 SUPPLEMENTARY APPENDIX. incidents fewer in number, and diminished hostility to the operation, it may be that on the whole it would promote the cause of vaccination, and secure, as its result, that the number of vaccinated persons would be greater than at present. Means, other than Vaccination, for diminishing the Prevalence of Small-pox ,- and how far such means could be relied on in place of Vaccination. Another question upon which we are asked to report is, what means, other than vaccination, can be used for diminishing the prevalence of small-pox ; and how far such means could be relied on in place of vaccination. The means, other than the inoculation of small-pox or cow-pox, which have been referred to by witnesses as being capable of diminishing the prevalence of small-pox, are such means as have been employed against infectious diseases generally ; they may be summarised as — (1) Measures directed against infection, e.g., prompt notification, isolation of the infected, disinfection, etc. ; (2) Measures calculated to promote the public health, the prevention of overcrowding in dwellings or on areas, cleanliness, the removal of definite insanitary conditions, etc. The principle underlying the practice of isolation with its accompany- ing machinery is obviously the very opposite of that which recommended the practice of inoculation ; it aims at exclusion of the disease, whereas inoculation aimed at universal acceptance by artificially " sowing " or " buying " the disease. Except in regard to the plague, our knowledge and practice of measures of isolation and quarantine against epidemics is of relatively recent growth. As the result of increased knowledge of the mode of propagation of infectious diseases, of greater sanitary activity, and under the stimulus of legislation, organised effort, more or less thorough, is now, in this as in other countries, directed against the spread of dangerous infectious diseases. Side by side with a vaccination system,, means of isolation, etc., have been successfully employed to check the spread of small-pox. . They have also been sometimes so employed in recent years in places where vaccination has fallen into disuse. It will be well to commence with a brief statement of the growth of our knowledge on the subject of isolation as a means of dealing with infectious or contagious diseases. We have already adverted to the fact that small-pox is highly contagious, and that contagion from those suffering from it is the means by which the disease is propagated. Although reference to infection appears in some of the Arabian writers,, the contagiousness of small-pox attracted little attention in this country and in western Europe until the eighteenth century. Sydenham (1624-89), though he refers to the contagiousness of small-pox, did not dwell upon the matter, and did not regard it as so important an element in the spread of the disease as some peculiar constitution of the atmosphere to which he attributed epidemics. Boerhaave was the first, at the commencement of the eighteenth century, distinctly to formulate the now generally accepted doctrine that small-pox arises only from contagion. In 1720, Mead drew up an elaborate system of notification, isolationr disinfection, etc., in view of a threatened invasion of the plague ; but no REPORT OF THE ROYAL VACCINATION COMMISSION. 689 attempt to deal with small-pox in a similar fashion appears to have been, made until the last quarter of the eighteenth century. This was in all probability largely due to the adoption of inoculation as the recognised defence against small-pox, and the acceptance of Sydenham's doctrine of epidemic causation may have exercised an influence in the same direction. No writer appears to have suggested methods of isolation, disinfection, etc., against small-pox prior to 1763. In that year Rast of Lyons published his " Reflections on Inoculation and Small-pox, and upon the means which . might be taken to deliver Europe from that malady." He maintained— (1) That small-pox was not a necessary and inevitable malady ; (2) That it arose only from contagion ; (3) That it resembled plague in most of its features. His conclusion was expressed in these terms : u I say, that to " deliver Europe from small-pox we must act upon principles directly " opposed to inoculation ; far from multiplying the contagion, we must " keep it away by taking the same precautions and employing the same " measures against that malady as have proved so successful against leprosy " and the plague." The earliest account of the practical employment of such means is from Rhode Island, U.S.A. Haygarth, on the authority of Drs. Moffat and Waterhouse, states that for many years prior to 1778 small-pox had been successfully prevented from becoming epidemic there by regulations for isolation of the infected on a neighbouring small island specially used for that purpose, and for quarantining infected vessels, destruction of infected clothing, etc. Moreover, inoculation was discouraged at Rhode Island, and those who wished to be inoculated had to go to some place away from the Island, and were not to return until there was no danger of their infecting others. A passage in Dimsdale's work on Inoculation, published in 1781, shows that in some towns of England pest-houses were beginning to be used for small-pox. In 1784 Haygarth, of Chester, published his " Inquiry how to prevent the Small-pox," and in 1793 "A Sketch of a Plan to exterminate the Small-pox from Great Britain." The great epidemic of small-pox at Chester in 1774, to which allusion has already been made, was the occasion of Haygarth's first attempts at organised dealing with epidemics of small-pox with a view to preven- tion. In his " Inquiry " he combated Sydenham's doctrine that epidemics are due to some occult condition of the atmosphere, and argued that small-pox was always spread by infection only. He further maintained that the variolous poison could be carried as an infection for a little distance only through the air, and " consequently that the small-pox may be " prevented by keeping persons liable to the distemper from approaching " within the infectious distance of the variolous poison till it can be "destroyed." These views led him, upon the return of an epidemic in 1777, to propose a plan for the prevention of the natural small-pox, and in 1778 a society was formed to carry out the plan in Chester. The plan consisted on the one hand of a general inoculation at people's homes at stated intervals, on the ground that the inoculated small-pox was far less fatal or injurious than the natural small-pox, and on the other hand of 44 (J90 SUPPLEMENTARY APPENDIX. " Rules of Prevention " based on Haygarth's views of infection. In the report of the Society, called shortly uThe Small-pox Society," dated September 1782, it is stated that in the four and a half years of its existence two general inoculations had been held, and that the deaths from small-pox had been greatly lessened. Great difficulties, however, were met with. " A large proportion of the inhabitants " refused inocula- tion, and a large proportion also, " being fearless, or rather desirous, that their children should be infected with the natural small-pox," refused to obey the Rules of Prevention. Hence, though the same report states that the example of Chester had been followed by Liverpool, where " a general inoculation was successfully executed in the autumn of 1781 and another in the spring of 1782," and in Leeds, where a general inoculation was held in 1781 and another proposed in 1782, with such success that the Royal College of Physicians in Edinburgh appointed a committee to inquire into " the modes of conducting the general inocula- tions of the poor " thus adopted in these places, the plan met with such difficulties that it was ultimately abandoned. It will be observed that a general inoculation was an essential part of the plan proposed and carried out in 1778-82 ; but, writing in 1784, Haygarth looked forward to being able ultimately to dispense with inoculation, and in the preface to his later edition, published in 1793, he states more definitely that the adoption of his Rules of Prevention without any general inoculation might exterminate small-pox in some country other than Great Britain. It must be remembered, however, that Haygarth entertained the opinion that the infection of small-pox could not be carried through the air above about half a yard, and that no one could -be infected by the clothes of a person visiting a small-pox patient provided that he kept beyond this distance from the patient. It is obvious that if this had been established the control of the disease by isolation would be a much simpler matter than it really is. In the Medico-Chirurgical Review for 1796 there appeared an account of a work by Dr. Faust, of Leipsic, entitled " An Essay on the Duty of a Man to separate persons infected with the Small-pox from those in " Health, thereby to effect the extirpation of that disease equally from •" the towns and countries of Europe," in which it was argued that the first person ill in a place is the only source from which all the rest, perhaps hundreds and thousands, become affected, and that if he were put immediately into a situation where he could not injure by contact those who had not had the disorder, the spread of the disease would be prevented. In the same Review for 1799 appeared an account of establishments for the extirpation of small-pox. The failure of inoculation to attain the desired end is referred to, and legislation is urged to facilitate isolation. It is further stated that in 179(3 the Prussian College of Physicians re- ported favourably to the King on the project, and that at Halberstadt it had been resolved to establish a house for the purpose. At Cote d'Or in France a similar plan had been tried with success. In 1 798 Jenner's " Inquiry " was published, and in the early years of this •century inoculation began to be discouraged ; for a while the prospects of REPORT OF THE ROYAL VACCINATION COMMISSION. 691 annihilating small-pox by vaccination appear to have superseded, in the minds of many, the plans of Haygarth and others. Some vaccinators, however, like Willan and Ring, still looked to methods of quarantine and to national and municipal regulations promoting isolation to exterminate the small-pox. It is worthy of notice, too, that Haygarth himself, in a letter quoted by Dr. Cappe of York in a communication to the London Medical ami Pltyxi&il Journal (vol. iv., p. 429), dated October 13th, 1800, remarked, " An introduction of the vaccine still more than of the variolous inocula- tion would effectually promote the great object of my publications/' Prior to the year 1866 there was no provision made by law for enabling sanitary authorities to establish hospitals for infectious diseases, and thus to promote the isolation of such cases. The only institutions of that description then existing were the result of private effort. So far as regards small-pox there was, practically speaking, no provision for its treatment by means of isolation. The Sanitary Act of 1866 empowered, though it did not compel, local authorities throughout England and Wales, Scotland and Ireland, to provide or to join in providing isolation hospitals for the use of the inhabitants of their districts. There was further legislation on the subject by the Public Health Act, 1875 ; the Public Health (London) Act, 1891 ; the Public Health (Scotland) Act, 1867 ; and the Public Health (Ireland) Act, 1878, into the details of which it is not necessary to enter. The most recent Act relating to the matter is the Isolation Hospitals Act of 1893, which applies to the small towns and rural districts of England and Wales. St(tmj)ing-out System in Leicester suffered severely from small-pox in 1872, 346 deaths having been registered as caused by it. Two deaths from that disease occurred in 1873, but no other until 1877, when there were six, and one in the following year. The next year in which small-pox deaths were registered was 1881. There were two in that year, and five and three in the follow- ing years. Xo other death took place until 1892 and 1893, in which years the fatal cases numbered 21. Prior to 1875 the vaccination laws were well observed in Leicester. In that year the number of children born who were unaccounted for \\a< only some 4 per cent. Since then there has been, as we have seen, a marked and progressive decline in the number of vaccinations, especially since 1883, until at the present time 80 per cent, of the children born remain unvaccinated. The borough hospital for infectious diseases was erected in IK 71 -2 outside the town ; though within the last few years houses have been built in proximity to it. It appears to have been with Dr. Crane, the Medical Officer of Health in 1875, that the quarantining the inmates of an infected house, in addition to isolating the patient, originated. His successor, Dr. Johnston, established it in 1877 as a regular system. He was aided in this, after 1879, by the notification of infectious diseases then rendered compulsory by a private Act which Leicester, anticipating 092 SUPPLEMENTARY APPENDIX. most other towns, obtained in that year. Dr. Johnston reported that up to 1884 the spread of small-pox from imported cases had been arrested in 20 instances by the means he adopted. His successor, Dr. Tomkins, though, like his predecessors, regretting the increasing disuse of vaccination, bore testimony in his annual reports* to the efficacy of the measures adopted in Leicester, and expressed his opinion that had such a system been in force at Sheffield in 1887 it would not have suffered in the way it did. In 1892 small-pox became prevalent in different parts of England, especially in Lancashire and Yorkshire. Many of the large provincial towns suffered, and Leicester amongst them. There were, in 1892-3, 357 cases of small-pox in Leicester, of whom 21, or 5*8, died ; 193 households were invaded, containing 1234 persons. The first importation was by a tramp, whose disease, passing unrecognised, caused infection at a common lodging-house and at the workhouse. Eleven other importations of the disease by tramps occurred in the course of 1892-3. Leicester suffered less than many of the other large towns which have been invaded by small-pox during recent years, both in the number of cases and in the number of deaths. In connection with this, however, a point to which we have already called attention must be borne in mind. The disease was remarkably slight there in its fatality, even as regards those who, by reason of their age, could not be affected by the change of practice in relation to vaccination. Dr. Priestley, the Medical Officer of Health, claims, in his report to the Sanitary Committee for 1893, that it was by reason of the -energetic methods adopted that the disease had been prevented running riot through the town. His claim may be well founded. At all events, the experience of Leicester affords cogent evidence that the vigilant and prompt application of isolation, etc., even with the defects which were brought to light during the recent epidemic, is a most powerful agent in limiting the spread of small-pox. It is true that the system and appliances which appeared adequate for some years failed to prevent a serious outbreak of small-pox in 1892-3. We think its value was none the less real. Stamping-out System in London. In the Report of the Royal Commission of 1881, already alluded to, suggestions were made with regard to notification and isolation which have since been largely carried into effect. As we have said, it was con- sidered proved that the existing small-pox hospitals had caused a spread of the disease in their neighbourhood. We cannot but think that this may in some measure account for the greatly increased mortality from small-pox in London during the 1871-72 epidemic as compared with the rest of the country. It is true that the statistics relating to England and Wales outside the Metropolis include those of other large towns where the same evil was present ; but it probably did not exist there in so aggravated a form, and the effect may be neutralised by the statistics relating to smaller towns and rural districts with which they are combined. This idea has been suggested to us, as the result of the inquiry, how it has come about that whilst the Metropolis, in the decennium 1867-76, and again REPORT OF THE ROYAL VACCINATION COMMISSION. 693 down to 1885, compared so unfavourably with the rest of the country, the condition has since that date become so entirely changed ? We think it is impossible to attribute this change to vaccination. There is no reason to suppose that the position of the Metropolis in respect to vaccination has, since the year 1885, become superior to the rest of England and Wales : rather the other way, as the decrease in infantile vaccination has been greater during the last few years than in the rest of England and Wales. The change, therefore, must be due to some other cause. The hospitals which, in the opinion of the Commissioners, were propagating the disease in their neighbourhood, were in operation down to July 1882, when their Report was made. In 1877 and 1878, and again in 1881, small-pox was epidemic in London to a considerable extent. We have stated in detail in paragraph 471 * the steps which were taken by the Metropolitan Asylums Board in consequence of the recommenda- tions of the Royal Commission. It will be seen that the intra-urban hospitals still continued in use, and that complaints were made in 1884 that they were spreading small-pox in their vicinity, although the number in each of them was not allowed to exceed 50. In October 1884 this number was reduced to 25. It was not, however, until 1885 that the system now in operation was inaugurated, and all cases of small-pox were treated in hospital ships. It is impossible not to be struck with the fact that it is since the year 1885 that the Metropolis has presented so satisfactory an aspect as regards small -pox mortality. The facts to which we have been calling attention certainly seem to point to the conclusion that this has been due to a system of isolation, well organised and administered, the beneficial effect of which is no longer neutralised by a spread of the disease from the hospitals in which the isolation is carried out. Upon the whole, we think the experience of London affords cogent evidence of the value of a sound system of isolation in checking the spread of small-pox. Stamping-out Si/stem 'in Anstrnlia. The experience of isolation systems in Australia is interesting and worthy of special notice, because whilst in this country the quarantining of persons who have come in immediate contact with those suffering from small-pox has only been possible with the consent of the persons whom it was proposed to subject to quarantine, in Australia their removal to a place of isolation has been made compulsory. Australia, by virtue of its geographical position, and the consequent separation by long sea voyage from infected ports, enjoyed for a long time a sort of natural isolation. Thus, Hirsch, in his " Historical and Geographical Pathology," vol. i., pp. 133-4 (1881), remarks : — " The continent of Australia up to 1 838 had enjoyed an absolute " immunity from small-pox ; towards the end of that year the disease " appeared at Sydney, having been imported probably from China ; it " lasted, however, only a short time, and remained absent from the " continent until 18G8. In that year it was introduced into Melbourne by " a ship, and again it spread only to a slight extent, and quickly died out. * Final Report. 694 SUPPLEMENTARY APPENDIX. " By a rigorous inspection of ships on their arrival, it has been found " possible to prevent subsequent importations, a notable instance of pre- " vention having occurred in 1872. Tasmania has hitherto quite escaped " the disease ; so also has New Zealand, where an importation of it in " 1872 was prevented by strictly isolating a vessel that had arrived with '• small-pox on board." In New South Wales, Dr. MacLaurin, who has been President of the Board of Health since 1889, informed us that the Government act on the assumption that small-pox is an exotic disease, and that every case must have come from outside the colony, and it is therefore dealt with under a quarantine Act of William IV., originally instituted for dealing with cholera. By an Act passed in 1882, notification of small -pox was made compulsory on medical men and householders under heavy penalties. At Sydney notification of small-pox is followed up by the compulsory removal of the patient and all persons who have been in the house with the patient to the quarantine station at North Head. This station is 670 acres in extent, and situated on the peninsula at the mouth of Sydney Harbour, and is seven miles from the Health Office, with which there is telephonic and telegraphic communication. The persons are conveyed to the station by a steamboat comfortably fitted expressly for the purpose, and no difficulty has been experienced in effecting their removal. It was, in Dr. MacLaurin's opinion, by carrying out this practice of isolation and quarantine that " the epidemic of 1881-82 was suppressed," and small-pox " has 'never become epidemic since this plan has been adopted." The persons who have been in the house with the patient are detained 21 days in quarantine from the date of the last possible contagion. Should a case of small-pox arise among them, those who had been in contact with such infected person would be detained for a further period of 21 days, and so on. To facilitate this, the exposed persons are distributed in separate groups within the station. They are allowed to receive letters or parcels, etc., and a telegraph operator is employed, " whose special business it is " to work the telegraph at their request." " Reasonable compensation is " given by the Government for loss ; " and there are heavy penalties under the original Act whereby the quarantine is secured. The station is, accord- ing to Dr. MacLaurin, " a pleasant place to stay in, and everything is done " that can be done to make the people comfortable : they have nothing " whatever to do, and are free from all care, and they can spend the day " pleasantly enough ; but they do not like it." No one, however, raises any objection to the Sydney system : "the people are all very sensible about it." In all Australian towns the same system is carried out as strictly, with the result that there was not a case of small-pox in Australia on February 5th, 1890 ; and Dr. MacLaurin is of opinion that the risk of dying of small-pox in Australia is smaller than in any other part of the world. As regards vaccination : — In New South Wales it is very little practised ; there is no compulsory Act ; and though medical opinion is in favour of it, an opinion shared by Dr. MacLaurin, it is not likely that a compulsory Vaccination Act could be passed or would be tolerated. The proportion of young persons in New South Wales who are not vaccinated is accordingly very krge : probably much more than half of those under REPORT OF THE ROYAL VACCINATION COMMISSION. 695 ten years of age are unvaccinated. Although Dr. MacLaurin favours vaccination and respects it highly, he is satisfied that the system of isola- tion as supervised by him is perfectly successful. As President of the Board of Health he considered it his business to produce extinction of the disease ; he does not consider vaccination a sufficiently absolute protection for such purpose ; and he is " fully of opinion that the only way in which " you can bring to an end an outbreak of small-pox, that is to say, bring " it under control, and not leave it to work itself out, is by notification " and isolation. Of course, in any small community, if you let the disease " in it will work itself out in time, because all the susceptible people will " have had it ; but the only way in which you can absolutely control an " epidemic of small-pox is by a system of notification and isolation." Small-pox has never been epidemic in Western Australia. Only one case has occurred within the last 31 years, and that was an imported one ; quarantine was carried out, and no infection occurred ; the immunity from the disease is mainly at least due to isolation. Before 1879 vaccination was not generally practised — a great majority of those born in the colony were unvaccinated ; in that year a compulsory Vaccination Act was passed in consequence of Sir H. Ord and Dr. Waylen's representations, and in consequence of reports of small-pox in other colonies, and not on account of the existence of small-pox in Western Australia. In Tasmania there was a compulsory vaccination law, but it was found to be inoperative because no one was appointed to conduct the prosecu- tions, and it has now fallen into desuetude. The same system of isolation and quarantine is exercised as in the other Australian colonies. Small-pox was for the first time introduced into Tasmania in 1887, and although preparations for isolation were inadequate, the disease was soon stamped out. Communication between Launceston and Melbourne was temporarily suspended, and to this precaution the non-invasion of Victoria was attributed. The particulars of this, the first introduction of small-pox into Tasmania during the history of that colony, are to be found in a report to the Central Board of Health by Mr. A. Mault, dated Novem- ber 17th, 1887. The origin of the outbreak is not clear, but it was presumed to have been imported, probably by a ship from China, into Launceston. The earliest case reported to the Local Board of Health was on September 23rd, though it appears that earlier cases had passed unnoticed, or had been notified as measles. Thirty-three cases in' all occurred, every one of which was traced to direct infection from the first case. By September 27th a temporary hospital had been erected, and thither patients and suspects to the number of 72 were removed. The last case appeared on October 13th. Other persons who had been to the infected houses were isolated in their houses and watched. Only four of the 47 persons quarantined at the station were attacked. The clothing was burnt, and very thorough disinfection of the infected houses was carried out, and the dead were interred in a special cemetery. The other colonies were communicated with, and quarantine, at first unduly rigid and after- wards relaxed, was practised against ships proceeding from Tasmania. Although vaccination had been nominally compulsory in Tasmania, it was estimated that two-fifths of the population were unvaccinated. 696 SUPPLEMENTARY APPENDIX. Suggested Stamping-out System In the United Kingdom. We have no difficulty in answering the question, what means other than vaccination can be used for diminishing the prevalence of small-pox ? We think that a complete system of notification of the disease, accom- panied by an immediate hospital isolation of the persons attacked, together with a careful supervision, or, if possible, isolation for sixteen days of those who had been in immediate contact with them, could not but be of very high value in diminishing the prevalence of small-pox. It would be necessary, however, to bear constantly in mind, as two conditions of success : first, that no considerable number of small-pox patients should ever be kept together in a hospital situate in a populous neighbourhood : and secondly, that the ambulance arrangement should be organised with scrupulous care. If these conditions were not fulfilled, the effect might be to neutralise or even do more than counteract the benefits otherwise flowing from a scheme of isolation. When we turn to the other branch of the inquiry, how far such means could be relied on in the place of vaccination, we find ourselves involved in questions of a much more complicated nature. We have little or no experience to fall back upon. The experiment has never been tried. The nearest approach to a trial of it has probably been in Australia. But even in the parts of that country to which we have alluded the population has not been entirely unvaccinated, though there has been a large unvaccinated class amongst it. Moreover, in applying the experience of Australia to this country, two things must be borne in mind. In the first place small-pox has only appeared from time to time, introduced from without at one or other of the ports of the country, and the several colonies of which Australia is composed are of great territorial extent, with few large centres of population. In this country small-pox is always present in some part of it. There has not been a single year without several deaths from the disease. Large centres of population are numerous, and the intercourse between them constant. In the several colonies of Australia the number of ports is not great, the vessels which enter them are comparatively speaking not numerous, and the ports from which they arrive are many days' voyage distant ; and there are careful arrangements for quarantining vessels to exclude disease. The shipping which enters English ports is of vast quantity, and passengers are brought m large numbers from the continent of Europe not only daily, but it may almost be said hourly ; the voyage, too, is but brief. The other matter to be remembered is, that part of the Australian system is the compulsory removal to quarantine for 21 days of those who have been in the house with the patient, in addition to the transfer of the patient himself to a lospital. There can be no doubt that such a system, if completely carried out, would be of the highest efficacy. But it is obvious that in this ountry the practical difficulties of working such a scheme in the large towns would be really insuperable, to say nothing of the difficulty of procuring legislative sanction for it. REPORT OF THE ROYAL VACCINATION COMMISSION. 697 Value <>f We can see nothing, then, to warrant the conclusion that in this country vaccination might safely be abandoned, and replaced by a system of isolation. If such a change were made in our method of dealing with small-pox, and that which had been substituted for vaccination proved ineffectual to prevent the spread of the disease (it is not suggested that it could diminish its severity in those attacked), it is impossible to contem- plate the consequences without dismay. To avoid misunderstanding, it may be well to repeat that we are very far from underrating the value of a system of isolation. We have already dwelt upon its importance. But what it can accomplish as an auxiliary to vaccination is one thing : whether it can be relied on in its stead is quite another thing. Even admitting fully the protective effect of vaccination, it does not, in our opinion, diminish the importance of measures of isolation or dis- pense with their necessity. We think that steps should be taken to secure a more general provision for the isolation of small-pox patients than exists at present. We have already called attention to the fact that mischievous results are likely to follow the use as a small-pox hospital of a building situate in a populous place. We think that wherever it is placed it should have sufficient space around it to enable the sanitary authority to add rapidly to the accommodation by the erection of tem- porary buildings. Compulsory Pt'oriximt <>f Isolation HojipitcUs. Sanitary authorities are now sometimes reluctant to provide isolation hospitals. We think that, on a petition by a prescribed number of the ratepayers in a sanitary district, the Local Government Board, if satisfied that the hospital accommodation ought to be provided, should have power to make an Order for such provision. y Notification of >>'/ We think that notification of small-pox should everywhere be compul- sory, and, whenever the disease showed a tendency to become epidemic, a notice should be served by the sanitary authority upon all persons in the neighbourhood who would be likely to come within the reach of con- tagion, urging them to submit to vaccination or re-vaccination, as the case might be, if they had not been recently successfully vaccinated or re- vaccinated ; and attention should be called to the facilities afforded for their doing so. Attention should also be called to the importance of avoiding contact with persons suffering from the disease, or coming into proximity to them, and of avoiding contact with any person or thing which may have become infected. It is important to notice that, even where vaccination has been neglected, there is great readiness to submit to it in the presence of a threatened epidemic ; a large number of vacci- nations are then obtained willingly and without opposition. Whenever 698 SUPPLEMENTARY APPENDIX. a sanitary authority has received notification of a case of small-poxr we think the fact should be at once communicated to the vaccination authority of the district in which the case of the disease has occurred. Regulations as to Tramps, Inmates of Lodging-houses, etc. Our attention has been drawn to the circumstance that outbreaks of small-pox have not unfrequently had their origin in the introduction of the disease to common lodging-houses by tramps wandering from place to place. In view of this we make the following recommendations : — (i.) That common shelters which are not now subject to the law relating to common lodging-houses should be made subject to such law. (ii.) That there should be power to the local authority to require medical examination of all persons entering common lodging- houses and casual wards to see if they are suffering from small-pox, and to offer a reward for prompt information of the presence of the disease. (iii.) That the local authority should have power to order the keeper of a common lodging-house in which there has been small- pox to refuse fresh admissions for such time as may be required by the authority. (iv.) That the local authority should be empowered to require the temporary closing of any common lodging-house in which small-pox has occurred. (v.) That the local authority should have power to offer free lodgings to any inmate of a common lodging-house or casual ward who may reasonably be suspected of being liable to convey small-pox. (vi.) That the sanitary authority should give notice to all adjoining sanitary authorities of the occurrence of small-pox in common lodging-houses or casual wards. (vii.) That where the disease occurs, the Public Yaccinator or the Medical Officer of Health should attend and vaccinate the inmates of such lodging-houses or wards, except such as should be unwilling to submit themselves to the operation. Relaxation of the Vaccination Law. After careful consideration and much study of the subject, we have arrived at the conclusion that it would conduce to increased vaccination f a scheme could be devised which would preclude the attempt (so often a vain one) to compel those who are honestly opposed to the practice to submit their children to vaccination, and, at the same time, leave the law o operate, as at present, to prevent children remaining unvaccinated owing to the neglect or indifference of the parent. When we speak of honest opposition to the practice, we intend to confine our remarks to * m which the objection is to the operation itself, and to exclude REPORT OF THE ROYAL VACCINATION COMMISSION. 699 cases in which the objection arises merely from an indisposition to incur the trouble involved. We do not think such a scheme impossible. It must of course be a necessary condition of a scheme of this description that it should be such as would prevent an objection to the practice being alleged merely as an excuse to save the trouble connected with the vaccination of the child. We may give the following as examples of the methods which might be adopted. It might be provided that if a parent attended before the local authority and satisfied them that he entertained such an objection, no proceedings should be taken against him. Or, again, a statutory declaration to that effect before any one now authorised to take such declaration, or some other specified official or officials, might be made a bar to proceedings. We do not think it would be any real gain to parents who had no conviction that the vaccination of their children was calculated to do mischief, to take either of these steps rather than submit them to the operation. It is in England that the point we have been recently discussing is of most practical importance, but if our suggestion were adopted the change should, of course, be made in all parts of the United Kingdom. (Signed) HERSCHELL. JAMES PAGET. CHARLES DALRYMPLE. W. GUYER HUNTER. EDWIN H. GALSWORTHY. JOHN S. DUGDALE. M. FOSTER. JONATHAN HUTCHINSON. FREDERICK MEADOWS WHITE. SAM. WHITBREAD. JOHN A. BRIGHT. BRET INTK, August 1890. Secretary. The undersigned do not find themselves able to go so far in recom- mending relaxation of the law as is implied. We think that in all cases- in which a parent or guardian refuses to allow vaccination, the person so- refusing should be summoned before a magistrate, as at present, and that the only change made should be to permit the magistrate to accept a sworn deposition of conscientious objection, and to abstain from the infliction of a fine. We are also of opinion that, in spite of the difficulties as set forth in paragraph 533,* a second vaccination at the age of twelve ought to be mad<* compulsory. W. GUYER HUNTER. JONATHAN HUTCHINSON. * Of the Final RepDrt. 700 SUPPLEMENTARY APPENDIX. We the undersigned desire to express our dissent from the proposal to retain in any form compulsory vaccination. We cordially concur in the recommendation that conscientious ob- jection to vaccination should be respected. The objection that mere negligence or unwillingness on the part of parents to take trouble might keep many children from being vaccinated would be largely, if not wholly, removed by the adoption of the Scotch system of offering vaccination at the home of the child, and by providing for medical treatment of any untoward results which may arise. We therefore think that the modified form of compulsion recommended by our colleagues is unnecessary, and that in practice it could not be carried out. The hostility which compulsion has evoked in the past toward the practice of vaccination is fully acknowledged in the Report. In our opinion the retention of compulsion in any form will in the future cause irritation and hostility of the same kind. The right of the parent on grounds of conscience to refuse vaccination for his child being conceded, and the offer of vaccination under improved conditions being made at the home of the child, it would in our opinion be best to leave the parent free to accept or reject this offer. SAM. WHITBREAD. JOHN A. BRIGHT. W. J. COLLINS. J. ALLANSON PICTON. Aote. — Dr. Collins and Mr. Picton sign the above note of reservation, tb'itif/h //„,/ 1,,1-re not signed the Report. A statement of their grounds of d'txxi;itf,'oin, Ike Report will be found in the form of an Appendix (65 payei) to the Report. They make the following recommendations : — In accordance with the sub-head No. 2 of the reference to the -Commission, we would suggest the following as the means other than vaccination which should be employed for protection of a community -from small-pox : — 1. Prompt notification of any illness suspected to be small-pox. Improved instruction in the diagnosis of small-pox. }. A hospital, suitably isolated, of adequate accommodation, in per- manent readiness, and capable of extension if required. No other disease to be treated at the same time in the same place. 3. A vigilant sanitary staff ready to deal promptly with first cases, and if necessary to make a house-to-house inspection. The medical officer of health to receive such remuneration as to render him independent of private practice. 4. Prompt removal to hospital by special ambulance of all cases which cannot be properly isolated at home. Telephonic com- munication between Health Office and hospital. REPORT OF THE ROYAL VACCINATION COMMISSION. 701 5. Destruction of infected clothing and bedding, and thorough dis- infection of room or house immediately after removal of the patient. 6. Daily observation (including, where possible, taking the tempera- ture and inspection for rash) of all persons who have been in close contact with the patient during his illness ; such super- vision to be carried out either in quarantine stations (away from the hospital) or at their own homes. 7. Closure of schools on the occasion of the occurrence of small-pox among the scholars or teachers. 8. Hospitals and quarantine stations to be comfortable and attractive, and so administered as to secure the confidence of the public. Hospital treatment to be free to all classes, and compensation to be paid to those detained or otherwise inconvenienced in the public interest, at the public expense. 9. Tramps entering casual wards to be medically inspected, their clothing to be disinfected, and bath provided. The measures for detection and isolation of small-pox in common lodging- houses suggested in section 507 of the Report to be carried out. 10. International notification of the presence of small-pox, and special vigilance at seaports in communication with infected places, after the plan adopted in the case of cholera. 11. Attention to general sanitation — prevention of overcrowding. abundant water supply, and frequent removal of refuse. They conclude as follows : — . We believe the methods of isolation of the infected, disinfection, and the observance of strict cleanliness, are both more successful and more legitimate methods for the State to encourage. They have the advantage of applying the preventive only where it is required ; and they do not necessitate an operation upon the person of every healthy individual. We therefore recommend that the law be amended by the repeal of the compulsory clauses of the Vaccination Acts. But in consideration of the prevalent belief in the value of vaccination as a prophylactic for an indefinite period, we suggest that in other respects the law should be left as it is, subject, however, to such modifications as are recommended for the diminution of attendant risks. The precedent established in the case of the abolition of compulsory church rates might be followed with advantage. In that case all machinery for laying and collecting the rate was lef Antact though the power of enforcement was taken away. The effect of our recommendation, if adopted, would be that vaccination would continue to be provided as at present for those who desire to avail themselves of it, but efforts to secure vaccination would be limited to moral influence— in a word, the whole country would be in the position of those unions in which the guardians have abandoned compulsion. The grounds on which we object to the enforcement of vaccination by penalties necessarily lead us to object to any method of indirect 702 SUPPLEMENTARY APPENDIX. compulsion. We regard as both inexpedient and unjust exclusion from any branch of the public service because of the refusal to submit to vaccination or re- vaccination. The injustice is perhaps most severely felt in the case of candidates for employment as pupil-teachers in public elementary schools. There are now districts in which, owing to the general opposition to vaccination, scarcely a girl or boy can be found who is legally eligible, and candidates have to be brought in at great incon- venience from surrounding districts. The existence of an exceptional case or cases in which such rejected candidates have at some time after- wards taken small-pox is in our view no justification for the continuation of this grievance. Statistics furnished to the Commission prove that large numbers of vaccinated or re-vaccinated persons have taken the disease ; and we are not aware of any evidence to show that vaccinated pupil-teachers have any special immunity. If our recommendations were carried out, the danger of contagion would be greatly diminished in schools, as elsewhere. On the whole, then, while there is much in the report of our colleagues from which we dissent, and we have accordingly abstained with reluctance from adding our signatures to theirs, we are at one with them in holding that it is unwise to attempt to enforce vaccination on those who regard it as useless and dangerous. We, however, go further, and agree with our colleagues, Mr. Whitbread and Mr. Bright, that it would be simpler and more logical to abolish compulsory vaccination altogether. INDEX 703 INDEX. Abbe's condenser, 74 Aberration, chromatic, 67 — spherical, 67 Abscess, 173 Achorion Schonleinii, 584 Actinomycosis, 413 — 447 — bovis, 434 — cattle to cattle, 444 — cultivation of, 436 hominis, 431 • in cattle, 429 — in man, 426 transmission of, from man to lower animals, 443 Agar-agar, 625 Air, compressed, 24 examination of, 140 - Hesse's method, 141 - Koch's „ 141 Petri's „ 142 - Pouchet's „ 143 — Sedgwick's „ 143 Aitken's tubes, 129, 630 Alexines, 57 Alum carmine, 616 Amoeba coli, 610 Anaerobes, classified, 494 cultivation of, 130 — 133 Aniline water, 617 Animals Order, 1878, 455 Anthrax, 42, 183, 191—216 - bacillus of, 192, 197 in horses. 207 in swine, 203 origin and spread of, 198 preventive inoculation, 209 „ measures in, 199 Anthrax, stamping out system, 210 Antiseptics, 30 Antitoxins, 56, 57 — diphtheria, 58 — 62 septic infectious, 63 tetanus, 62 — typhoid, 64 Arthrospores, 19 Artificial fluids as media, 120 Ascococcus, 14 - Billrothii, 498 — citreus, 498 Ascomycetes, 584 Asiatic cholera. 360 Aspergillus albus, 588 clavatus, 588 flavescens, 588 — flavus, 586 fumigatus, 586 glaucus, 586 nidulans, 588 — niger, 587 — ochraceus, 588 — repens, 586 subfuscus, 588 B Babes' incubator, 633 Bacillus, 13, 488 — acidiformans, 498 — acidi lactici, 498 — aerogenes, 499 capsulatus, 499 — aerophilus. 499 — albus, 499 anaerobiescens, 499 • cadaveris, 499 — putidus, 499 05 45 706 INDEX. Bacillus allantoides, 499 allii, 499 alvei, 470 amylobacter, 502 amylozyma, 500 anaerobicus liquefaciens, 500 — anthracis, 192 aquatilis, 500 fluorescens, 500 graveolens, 500 — sulcatus, 500 arborescens, 500 argenteo-liquefaciens, 501 — phosphorescens, i<0l aurantiacus, 501 aureus, 501 beroliniensis indicus, 502 brassicae, 502 — brevis, 502 brunneus, 502 buccalis fortuitus, 502 — maximus, 502 minutus, 502 butyricus, 502, 503 cadaveris, 503 canalis capsulatus, 504 — parvus, 504 — candicans, 504 capsulatus, 504 mucosus, 504 — suis, 504 — carabiformis, 504 — carnicolor, 504 — carotarum, 505 cavicida, 505 — chromo-aromaticus, 505 Havaniensis, 505 circulans, 505 citreus cadaveris, 505 cloacag, 505 coeruleus, 505 — coli communis, 344 — similis, 506 — constrictus, 506 — coprogenes foetidus, 506 — parvus, 506 — crassus aromaticus, 506 sputigenus, 506 cuniculicida, 228 cuticularis, 506 — albus, 506 — cyaneo-fuscus, 507 Bacillus cyaneo-phosphorescens, 507 cyanogenus, 507, 508 cystiformis, 508 — delicatulus, 508 dentalis viridans, 508 dentriticus, 508 devorans, 508 - diffusus, 508 - diphtherias, 332—336 columbarum, 336 vitulorum, 509 dysodes, 509 endocarditidis capsulatus, 509- griseus, 509 enteritidis, 372 epidermidis, 509 — erysipelatis suis, 356 erythrosporus, 509 figurans, 510, 511 filiformis, 511 Havaniensis, 511 flavescens, 511 flavocoriaceus, 511 fluorescens aureus, 511 liquefaciens, 511 — longus, 512 minutissimus, 512 nivalis, 512 • non-liquefaciens, 512 • putidus, 512 — tenuis, 512 foetidus, 513 ozsenae, 513 — fulvus, 513 fuscus, 513 — limbatus, 513 — • gallinarum, 513 — gasoformans, 513 — glaucus, 513 gliscrogenus, 513 gracilis, 514 granulosus, 514 — graveolens, 514 guttatus, 514 haemorrhagic septicaemia, 231 — halophilus, 514 — Hansenii, 514 Havaniensis, 505 liquefaciens, 514 — helvolus, 515 heminecrobiophilus. 515 — hepaticus fortuitus, 515 INDEX. 707 Bacillus Hessii, 515 hyacinth! septicus, 515 — hyalinus, 515 hydrophilus fuscus, 515 ianthinus, 516 implexus, 516 — in acne contagiosa in horses, 516 in cancer, 516 incanus, 518 — in cholera in ducks, 516 - in choleraic diarrhoea, 516 — indicus, 518 indigogenus, 519 in diphtheritic disease of calves, 516 -- in erythema nodosnm, 517 in infectious disease of bees, 471 inflatus, 519 in fowl enteritis, 230 in gangrene, 517 in grouse disease, 517 in hog cholera, 517 in infantile diarrhoea, 517 in intestinal diphtheriain rabbits, 517 in jequirity infusion, 517 in measles, 517 in noma, 517 in potato rot, 517 in purpura heemorrhagica, 517 in putrid bronchitis, 518 — in "red-cod," 518 in rhinoscleroma, 518 • in saliva, 518 — in swine fever, 346-352 • erysipelas, 354 measles, 354 — inunctus, 519 invisibilis, 518 in whooping cough, 518 — iridescens, 519 iris, 519 lactericeus, 520 — lactis aerogenes, 519 — albus, 520 erythrogenes, 520 pituitosi, 520 — leporis lethalis, 520 lepra;, 407 leptosporus, 520 — limbatus acidi lactici, 520 limosus, 520 Bacillus liodermos, 521 liquefaciens, 521 communis, 521 magnus, 521 — parvus, 521 liquidus, 521 - litoralis, 521 - lividus, 524 - luteus, 524 — maidis, 524 - mallei, 452 megatherium, 524 — membranaceus amethystinus, 525 meningitidis purulentae, 525 mesentericus f uscus, 525 — ruber, 525 vulgatus, 526 inultipediculus, 522 muscoides, 522 — mycoides, 522 fa ^ roseus, 522 ^1» — neapolitanus, 522 necrophorus, 523 — nitrificans, 523 nodosus parvus, 523 — nubilus, 523 — ochraceus, 523 oedematis aerobicus, 523 — maligni, 220 — of Belfanti and Pascarola, 523 — of Colomiatti, 526 - of Fulles, 526 of Guillebeau, 526 of Letzerich, 526 — of Martinez, 526 - of Nocard, 526 — of Okada, 526 of quarter-evil, 217 — of Roth, 526 - of Sattler. 526 — of Schaffer, 527 of Scheurleu, 527 — of Schou, 527 — of septicaemia of buffaloes, 226 of guinea-pigs, 224 — • of mice, 225 of swine plague, 351 of Tommasoli, 527 of Utpadel, 527 — of Winogradsky, 527 ophthalmia, 190 ovatus minutissimus, 527 708 INDEX. Bacillus oxytocus perniciosus, 527 pestifer, 528 phosphorescens gelidus, 528 — indicus, 528 indigenus, 528 plicatus, 528 pneumonias crouposse, 233 pneumosepticus, 528 . polypiformis, 528 prodigiosus, 528 proteus fluorescens, 529 pseudo-diphtheriticus, 529 tuberculosis, 529 — pulpae pyogenes, 529 punctatus, 529 putrificus coli, 529 — pyocyaneus, 529 pyogenes fcetidus, 530 soli, 530 — radiatus, 530 aquatilis, 530 ramosus, 531 reticularis, 531 rhinoscleroma, 411 — rosaceus metalloides, 531 rubefaciens, 531 — rubellus, 531 — ruber, 531 — rubescens. 532 - rubidus, 532 sanguinis typhi, 532 — saprogenes, 532 scissus, 532 — septicaemia; haemorrhagicae, 231 — septicus, 532 acuminatus, 533 agrigenus, 583 keratomolaciae, 533 — ulceris gangraenosi, 533 vesicae, 533 sessilis, 533 — smaragdino-phosphorescens, 533 — smaragdtnus fretidus, 534 solidus, 534 — spiniferus, 534 spinosus, 534 • — stolonatus, 534 stolon if eras, 534 — striatus albus, 534 — flavus, 534 — subflavus, 535 subtilis, 535 Bacillus subtilis similans. 536 sulfureus, 536 superficialis, 537 — syphilis, 410 — tenuis sputigenus, 537 termo, 537 tetani, 457 thelassopbilus, 537 thermophilus, 537 — tremelloides, 537 — tuberculosis, 378 gallinarum, 402 — tumescens, 537 — typhi abdominalis, 342—346 murium, 359 — ubiquitus, 537 - ulna, 538 vacuolosis, 538 varicosus conjunctives, 538 — venenosus, 538 brevis, 538 — . invisibilis, 538 liquefaciens, 539 ventriculi, 539 vermicularis, 539 vermiculosus, 539 violaceus, 539 laurentius, 539 • virescens, 539 — viridis pallescens, 539 — viscosus, 540 Zurnianus, 540 Bacteriaceae, 480 Bacteria, chemical action on, 25 products of, 39 — composition of, 11 — distribution of, 29 - form of, 13 growth of, 23, 25 — in cattle plague, 296 in diphtheria of pigeons, 336 — in distemper, 358 — in foot and mouth disease, 300 in horse-pox, 312 — in liquids, 83 in louping-ill, 463 • • in measles, 283 — in pus, 176 — in rabies, 460 in scarlet fever, 262 in sheep-pox, 298 — in small-pox, 285 INDEX. 709 Bacteria in vaccine lymph, 324—326 — in yellow fever, 260 — microscopic examination of, 83 nitrifying, 27 — pathogenic, 27 saprogenic, 26 — stained, 85 unstained, 84 Bacterium aerogenes, 540 brunneum, 540 fusiforme, 540 gingivae pyogenes, 540 hyacinth!, 540 hydrosulfureum ponticum, 540 — litoreum, 540 luteum, 540 — merismopedioides, 541 navicula, 541 photometricum, 541 — synxanthum, 541 — termo, 541 tholoeideum, 541 — ureas, 541 — violaceum, 541 — Zopfii, 542 Beggiatoa alba, 542 roseo-persicina, 542 Bibliography, 639 Bilious fever, 183 Biondi's stain, 615 Bismarck-brown, 617 Blepharadenitis, 184 Blood-serum, 113, 114, 119 Borax carmine, 617 Botrytis Bassiana, 588 Bread paste, 118 Broncho-pneumonia, 183 Broth, 118 Bunge's method of staining flagella, 92 Camera lucida, 621 Cancer bodies, 610 Caoutchouc caps, 626 Caterpillars, disease of, 492 Cattle plague, 293—296 — tuberculosis of, :5st) Cerebro-spinal meningitis, 184 Chemical action on bacteria, 25 — disinfectants, 32—36 products of bacteria, 39 Chemiotaxis, 54 Chionyphe Carteri, 588 Cholera, 360 — bacteria of, 371 — comma bacilli of, 361 — 367 — diarrhoea from meat-poisoning, 371 — • diarrhoea in fowls, 373 — nostras, 370 — protective inoculation, 369 Ptomaines, 41 — spirillum of, 361 Chromogenic bacteria, 25 Cladothriceas, 480 Cladothrix, 479 — dichotoma, 543 — Forsteri, 544 — intricata, 544 — invulnerabilis, 544 Classification, 475, 487, 482, 481 Clostridium butyricum, 544 — fcetidum, 545 Coccaceaa, 480 Cocci, 485 Coccidia, 609 Comma bacilli, 361—367 Cover-glass preparations, 86 Cow-pox, 274—282, 312—324, 326 — 328 Crenothrix Kiihniana, 545 D Dacryocystis, 184 Damp chambers, 628 D'Arsonval's incubator, 631 Davaine's septicaemia, 228 De Bary's classification, 481 Decalcifying, 93, 615 Defensive proteids, 57 Deneke's comma bacillus, 367 Desiccators, <>::s Diarrhoea, choleraic, 371 Diphtheria, 330, 1 ^L' — antitoxin in, 58—62 - bacillus of, 332 — in milk, 338 in pigeons, 336 Loffler's stain for, 88 — Ptomaines of, 46 Diplococcus, 14 albicans arnplus, ."> 1 7 tardissimus. 547 710 INDEX. Diplococcus citreus conglomeratus, 547 — liquefaciens, 547 coryzae, 547 flavus liquefaciens tardus, 547 fluorescens fcetidus, 547 intercellularis meningitidis, 548 • luteus, 548 - pneumonias, 233—238 — of horses, 548 — roseus, 548 subflavus, 548 Distemper in dogs, 358 Drop-cultures, 120 Duck cholera, 230 Dysentery, 373 E Ebner's solution, 615 Egyptian ophthalmia, 190 Ehrlich's staining method, 89 Electricity, 24, 127 Embedding, 93, 615 Empusa muscae. 581 — radicans, 582 Endospores, 18 Enteric, 41, 340 Enzymes, 48 Eosin, 89, 617 Epidemic disease of deer and boars, 227 of ferrets, 358 — of mice, 358 Erysipelas, 185—189 Esmarch's roll-cultures, 113 Experiments on animals, 134, 394 Farrant's solution, 621 Favus, 584 Filter, hot water, 624 Flacherie, 472 Flagella, 17, 90 Fliigge's classification, 481 Foot rot, 464 Form of bacteria, 13 Foul-brood, 469 Fowl cholera, 228 — enteritis, 230 scab, 586 Friedlander's bacillus, 233 Fuchsine, 617 G Gangrene, 182 Gas chamber, 127 Gases produced by bacteria, 24 Gentian violet stain, 617 Giant cells, 376 Gibbes' solution, 618 Glanders, 451 — . bacillus of, 452 mallein in, 454 • — •— ptomaine, 48 Glycerine agar, 104 Glycerine gelatine, 615 Gonococcus of Neisser, 189 Gonorrhoea, 189 Gram's method, 88, 89, 97, 618 Grease, 303 Grouse disease, 230 H Haematococcus bovis, 548 Hasmatomonas carassii, 605 — cobitis, 603 Haematoxylin, 618 Hgematozoa, 589, 593, 599, 603 Hardening, 93, 615 Helicobacterium aerogenes, 549 Herpetomonas Lewisi, 599 Hessert's stain, 92 Historical introduction, 1 Horse-pox, 303, 312 Hot air and steam, 36, 623 Hot water filter, 624 Hueppe's classification, 482 — inspissator, 630 Hydrophobia, 459 Hyphomycetes, 581 Hypodermii, 581 Illumination, 75 Immersion system, 69 Immunity, 50 — 57 Impression preparations, 92 Incubators, 631 Infectious pleuro-pneumonia, 239, 247 Influenza, 247 — 249 Inspissators, 629 Involution forms, 15 Iodine, 618 Isolation of micro-organisms, 139 Israel's case, 628 INDEX. 711 K Klein's micrococcus of pneumonia, 238 Kleinenberg's solution, 616 Koch's comma bacillus, 361 postulates, 9 serum steriliser, 629 steam steriliser, 622 Leprosy, 406 bacillus of, 407 stamping out system, 409 Leptothrix, 480 • buccalis, 549 gigantea, 549 Leuconostoc, 549 Leukaemia, 184 Light, effect of, on bacteria, 24 Lister's flasks, 128, 630 Lithium carmine solution, 618 Lb'ffler's solution, 619 — stain, 88, 90 Louping ill, 462 bacillus of, 463 Lutesch's stain, 91 M Madura disease, 447 Magenta solution, 618 Malaria, 589 Malignant oedema, 220 Mallein, 454 Measles, bacteria in, 183, 283 Media, 99 Merismopedia, 14 Methyl violet, 619 Methylene blue, 618, 619 Micrococcus acidi lactici, 550 liquefaciens, 550 aerogenes, 551 agilis, 551 citreus, 551 — albus liquefaciens, 551 amylivorus, 551 — aquatilis, 551 • invisibilis, 551 auranticaus, 551 • botyogenus, 552 — candicans, 552 candidus, 552 carneus, 552 cerasinus siccus, 552 Micrococcus cereus albus, 178 flavus, 178 cinnabareus, 552 — citreus, 552 concentricus, 553 cremoides, 553 crepusculum, 553 cumulatus tenuis, 553 — endocarditidis rugatus, 553 — fervidosus, 553 — Finlayensis, 553 — flavus desideus, 553 liquefaciens, 554 tardigradus, 554 fcetidus, 554 Freudenreichi, 554 — fuscus, 554 — gingivae pyogenes, 554 — gonorrhcese, 190 — Havaniensis, 555 in abscess in rabbits, 556 in Biskra button, 555 in gangrenous mastitis in sheep, 555 — in infectious pleuro-pneumonia, 555 — in influenza, 555 in pemphigus, 555-6 - in pneumonia, 236, 238, 556 — in py&mia in rabbits, 556 — in septicaemia in rabbits, 556 — in syphilis, 557 — in trachoma, 190 lactis viscosus, 557 — luteus, 557 — melitensis (Malta fever). 557 — ochroleucus, 557 — plumosus, 557 — pneumonias crouposae, 236 pyogenes tenuis, 557 rosaceus, 558 — rosettaceus, 558 — salivarus septicus, 558 — stellatus, 588 — tetragenus. 558 mobilis ventriculi, 558 subflavus, 558 versatilis, 558 — ureae liquefaciens, 559 — versicolor, 559 — violaceus, 559 viticulosus, 559 712 INDEX. Micrometer, 80 Microscope, 65, 612 Microsporon furfur, 586 Microtomes, 94, 614 Miescher's tubes, 609 Mildew, 581 Milk, 120 scarlatina, 265, 282 tubercular, 291 Miquel's bulbs, 129 Moist chambers and cells, 121 Moitessier's gas regulator, 634 Monads, 601 Monas Okenii, 560 - Vinosa, 560 Warmingii, 560 Mouse favus, 586 Movements of bacteria, 15, 24 Mucors, 583 Muller's fluid, 616 Myconostoc gregarium, 560 Myco-protein, 11 Nature of soil for bacteria, 23 Neelsen's solution, 89, 98, 619 Nicols' stain, 92 Nitromonas of Winogradsky, 560 Nutrient agar, 103 Nutrient gelatine, 100 O Oidium albicans, 586 lactis, 584 - Tuckeri, 584 Oil immersion, 70 Orseille, 619 Parietti's method, 148 Pasteur's apparatus, 130 Pathogenic bacteria, 27 Pebrine, 471 Pediococcus acidi lactici, 560 cerevisiae, 560 Penicillium glaucum. 588 Peronospora infestans, 582 Petri's method exam, of air, 142 Pfeiffer bodies, 610 Phagocytosis, 27, 55 Photogenic bacteria, 25 Photography of bacteria, 150, 168 Photo-micrographic apparatus, 621 Phycomycetes, 582 Phylaxins, 57 Picric acid, 619 Picro-carmine, 619 Picro-lithium carmine, 620 Pilobolus, 583 Plague, bacillus of, 250, 252 Plate cultivations, 106 Platinum needles, 83, 616 Pleuro-pneumonia, 239 Pneumo bacillus liquefaciens bovisr 242, 560 Pneumonia, 233—238 ptomaines, 48 Potash solution, 620 Potato medium, 115 — 117 Pouchet's aeroscope, 143 Preservation of preparations, 93 Protective inoculation, 49 - in anthrax, 209 in cholera, 369 — in rabies, 460 — - in sheep-pox, 298 in small-pox, 285, 293 in swine erysipelas, 336 Proteus capsulatus septicus, 560 — hominis capsulatus, 224 — in gangrene of the lung, 560 — microsepticus, 560 - mirabilis, 561 — • septicus, 561 sulf ureus, 561 vulgaris, 561 Zenkeri, 562 Pseudo-diphtheritic bacillus, 335 Pseudo-diplococcus pneumonias, 562. Psorosperms, 609 Ptomaines, 39 Pus, bacteria in, 176 Putrefactive bacteria, 26, 28 Pyaemia, 175 Quarter-evil. 217 Q B Eabies, 459—462 Kag-pickers' septicaemia, 224. Kefraction, 65 Kelapsing fever, 257 Eeproduction, 18 INDEX. via Kespiration of bacteria, 22 Rhabdomonas rosea, 562 Rhinoscleroma, 411 Ringworm, 585 S Saccharomyces acidi lactici, 580 albicans, 579 anomalus, 579 apiculatus, 578 aquifolii, 580 cerevisige, 577 conglomeratus, 578 — ellipsoideus, 577 exiguus, 578 glutinus, 579 Hansenii, 580 - ilicis, 580 Jorgensenii, 578 Ludwigii, 580 Marxianus, 580 membranaefaciens, 580 minor, 580 niger, 580 pastorianus, 578 pyriformis, 579 — rosaceus, 580 sphsericus, 579 Safranine, 620 Sapraemia, 175 Saprogenic bacteria, 26 Saprolegina, 582 Sapropbytic bacteria, 27 Sarcinae, 487 Sarcina alba, 562 — aurantiaca, 562 — Candida, 563 — flava, 563 - hyalina, 563 intestinalis, 563 — litoralis, 563 lutea, 563 mobilis, 563 — pulmonum, 563 Reitenbachii, 564 — rosea, 564 urinae, 5(54 ventriculi, 564 Scarlet fever, 261 Schizomycetes, 477 Scblosing's membrane regulator, 632 Sclavo's stain, 91 Septicaemia, 175 — of calves, 227 of Davaine, 224 — of guinea-pigs, 224 — of mice, 225 of rabbits, 228 — of rag-pickers, 224 Serum therapy, 56 Sheep-pox, 297 Small-pox, 182, 284-293, 326- Smut, 581 Soil, examination of, 144 " Sozins, 57 Sphaerotilus natans, 564 Spirilla, 495 Spirillum amyliferum, 564 anserum, 564 — : — attenuatum, 564 — aureum, 564 — choleras Asiaticae, 361 — choleroides, 565 concentricum, 565 — dentium, 565 — Finkler and Prior, 258 flavescens, 565 — flavum, 565 Giinther, 566 leucomelaneum, 565 linguas, 565 marinum, 565 — • Metchnikovi, 373 — . MiUer, 566 — nasale, 566 Neisser, 566 — • Obermeieri, 258 — plicatile, 466 — Renon, 566 — rosaceum, 566 — Rosenbergii, 566 — rubrum, 569 rufum, 566 rugula, 567 sanguineum, 567 — saprophiles, 567 — serpens, 568 — Smith, 566 — sputigenum, 568 — suis, 568 — tenue, 568 tyrogenum, 568 — undula, 568 volutans, 568 714 INDEX. Spirillum Weibel, 566 Spirochcetae, 596 Spiromonas Cohnii, 569 — volubilis, 569 Spontaneous generation, 3 .Spores, 18 Staining spores, 90 Staphylococcus, 14 — cereus albus, 178, 324 — flavus, 178 — pyogenes albus, 178 — aureus, 176, 324 — citreus, 178 — pyosepticus, 569 — salivarius pyogenes, 569 viridis flavescens, 569 Sternberg's bulbs, 128, 630 Streptococcus, 14 acidi lactici, 569 — albus, 569 — bombycis, 472 brevis, 569 cadaveris, 570 coli gracilis, 570 conglomerates, 570 — flavus desidens, 570 giganteus urethrae, 570 — — Havaniensis, 570 — in contagious mammitis in cows, 570 — in progressive necrosis in mice, 571 — in strangles, 571 — liquefaciens, 571 — mirabilis, 571 — of Bonome, 571 — of erysipelas, 185, 189 of Manneberg, 571 — perniciosus psittacorum, 572 — pyogenes, 178-184 radiatus, 572 septicus, 572 — liquefaciens, 572 — vermiformis, 572 Streptothrix, 496 — actinomycotica, 431 — albus, 572 — asteroides, 573 — aurantiaca, 573 carnea, 573 — chromogena, 573 • farcinica, 573 Streptothrix Forsteri, 573 — Hoffmanni, 573 liquefaciens, 573 — Madurae, 449 musculorum suis, 573 odorifera, 573 violacea, 573 Substage condenser, 74 Suppuration, 48 Surgical fever, 182 Surra, 593 Swine erysipelas, 354 - fever, 46, 227 • measles, 354 Syphilis, 183, 410 Tarichium megaspermum, 582 Temperature for bacteria, 24 Test-tube cultivations, 105 Tetanus, 457 ptomaine, 41 Thermo-regulators, 635 Tilletia caries, 581 Toxalbumins, 39 Trachoma, 412 Trenkmann's stain, 91 Trichomonas, 607 Tricophyton tonsurans, 585 Tripod levelling apparatus, 627 Tuberculosis, 43, 375 and meat, 397 and public health, 391 antitoxin, 64 - bacillus of, 378 giant cells in, 376 - in animals, 389, 394, 399, 400 in man, 387 Typhoid, 340 Typhus, 259 U Ulcerative endocarditis, 183 Undulina ranarum, 607 Uredo, 581 Urine as a medium, 120 Urobacillus Duclauxi, 573 Freudenreichi, 574 - Maddoxi, 574 - Pasteuri, 574 — Schutzenbergi, 574 INDEX. 715 Urocystis occulta, 581 Ustilago carbo, 581 Van Ermengen's stain, 91 Vegetable infusions as media, 120 Vesuvin, 620 Vibrio rugula, 574 W Warmstage, 123, 124 Water bath, 624 examination of, 145 Weigert's stain, 296 Wort gelatine, 104 ; Xylol, 616 Yeasts, 577 Yellow fever, 182, 259 Zooglea, 13 Zopf's classification, 480 1896. 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The entire book has been thoroughly revised. In addition to this general revision, special paragraphs on the following new matter have been introduced : Filamentous Keratitis, Blood-staining of the Cornea, Essential Phthisis Bulbi, Foreign Bodies in the Lens, Circinate Reti- nitis, Symmetrical Changes at the Macula Lutea in Infancy, Hyaline Bodies in the Papilla, Monocular Diplopia, Subconjunctival Injections of Germicides, Infiltration-Anaesthesia, and Sterilization of Collyria. Brief mention of Ophthalmia Nodosa, Electric Ophthalmia, and Angioid Streaks in the Retina also finds place. An Appendix has been added, containing a full description of the method of determining the corneal astigmatism with the ophthalmometer of Javal and Schiotz, and the rotations of the eyes with the tropometer of Stevens. The chapter on Operations has been enlarged and rewritten. A TEXT-BOOK OF HISTOLOGY, DESCRIPTIVE AND PRACTICAL. With 174 beautifully colored original figures. By ARTHUR CLARKSON, M. B., C. M., Edin., formerly Demon- strator of Physiology in the Owen's College, Manchester; late Demonstrator of Physiology in Yorkshire College, Leeds. Fur- nishing the student in one volume with all that is necessary for his equipment as an Histologist. Large 8vo. volume, strongly bound. Price, $6.00 net. The purpose of the book is to furnish the student of Histology, in one volume, with both the descriptive and the practical part of the science. In each chapter the structures of the various tissues and organs are first described ; the student is then taken tutorially over the illustrative specimens; finally, an appendix concludes each chapter with a short account of the methods of preparation. BULLETIN OF MEDICAL PUBLICATIONS: W. B. SAUNDERS, PHILADA. BOOKS -RECENTLY ISSUED. TEXT-BOOK OF THE PATHOGENIC BACTERIA. Specially written for students of medicine. By JOSEPH McFAR- LAND, M. D., Professor of Pathology and Bacteriology in the Medico- Chirurgical College of Philadelphia, etc. 359 pages, finely illustrated. Cloth. Price, $2.50 net. The book presents a concise account of the technical procedures necessary in the study of Bacteriology. It describes the life-history of pathogenic bacteria, and the pathological lesions following invasions. The illustrations have been gathered from standard sources, and com- prise the best and most complete aggregation extant. " The author has rendered a great service to the profession in bringing out this work at this opportune time." — The American Therapist, Aug.. 1896. PRACTICAL POINTS IN NURSING. For Nurses in Private Practice. By EMILY A. M. STONEY, Superintendent of the Training-school for Nurses, Carney Hospital, South Boston. 456 pages, handsomely illustrated with 73 engravings and 9 colored and half-tone plates. i2mo. Cloth. Price, $1.75 net. A vade mecum for the private nurse, and an efficient teaching-book for training-schools. Instructions for quickly improvising needed sick- room appliances constitute a valuable feature of the book. The Appendix contains, in compact form, much information that will be of great value to the nurse, including General Rules for Feeding the Sick ; Recipes for Invalid Foods and Beverages ; Tables of Weights and Measures ; Table for Computing Date of Labor ; List of Abbre- viations; Dose-List; and a Glossary of 64 pages. " The author's ^tyle is most agreeable, and she handles her subject in a way that clearly shows her complete familiarity with it." — New York Polyclinic, Aug. 15, 1896. " There are few books intended for non-professional readers which can be so cor- dially endorsed by a medical journal as can this one." — Therapeutic Gazette, Aug. 15, 1896. " This is a well-written, eminently practical volume." — American Journal of Obstetrics, Aug., 1896. THE PICTORIAL ATLAS OF SKIN DISEASES AND SYPHILITIC AFFECTIONS (American Edition). Translation from the French. Edited by J. J. PRINGLE, M. B., F. R. C. P., Assistant Physician to, and Physician to the depart- ment for Diseases of the Skin at, the Middlesex Hospital, London. Photo-lithochromes from the famous models of dermatological and syphilitic cases in the Museum of the Saint-Louis Hospital, Paris, with explanatory wood-cuts and text. In 12 Parts, at $3.00 per Parts Parts I. to IV. now ready. " The plates are beautifully executed." — JONATHAN HUTCHINSON, M. D. (London Hospital). BULLETIN OF MEDICAL PUBLICATIONS: W. B. SAUNDERS, PHILADA. JUST PUBLISHED. A TEXT-BOOK OF BACTERIOLOGY, including the Eti- ology and Prevention of Infective Diseases and an ac- count of Yeasts and Moulds, Hsematozoa, and Psoro- sperms. By EDGAR M. CROOKSHANK, M. B., Professor of Comparative Pathology and Bacteriology, King's College, London. A handsome octavo volume of 700 pages, illustrated with 273 engravings in the text, and 22 original and colored plates. Price, $6.50 net. This book, though nominally a Fourth Edition of Professor Crook- shank's " MANUAL OF BACTERIOLOGY," is practically a new work, the old one having been reconstructed, greatly enlarged, revised through- out, and largely rewritten, forming a text-book for the Bacteriological Laboratory, for Medical Officers of Health, and for Veterinary In- spectors. A MANUAL OF PHYSIOLOGY, with Practical Exer- cises. For Students and Practitioners. By G. N. STEW- ART, M. A., M. D., D. Sc., lately Examiner in Physiology, University of Aberdeen, and of the New Museums, Cambridge University; Professor of Physiology in the Western Reserve University, Cleveland, Ohio. Handsome octavo volume of 800 pages, with 278 illustrations in the text, and 5 colored plates. Price, Cloth, $3.50 net. In this book formal exposition has been interwoven with practical work, in the manner shown by the experience of the Author to be best suited to the needs and opportunities of the student. The systematic portion of the book is so arranged that it can be used independently of the practical work, and aims at being itself a complete exposition of the subject, adapted to the requirements of the student of medicine. " It will make its way by sheer force of merit, and amply deserves tc do so. It is one of the very best English text-books on the subject." — Lancet. "Of the many text-books of physiology published, we do not know of one that so nearly comes up to the ideal as does Prof. Stewart's volume."— British Medical Journal. WATER AND WATER SUPPLIES. BY JOHN C. THRESH, D. Sc., M. B., D. P. H., Lecturer on Public Health, King's Col- lege, London; Editor of the "Journal of State Medicine," etc. I2mo, 438 pages, illustrated. Handsomely bound in Cloth, with gold side and back stamps. Price, $2.25 net. This work will furnish any one interested in public health the infor- mation requisite for forming an opinion as to whether any supply or proposed supply is sufficiently wholesome and abundant, and whether the cost can be considered reasonable. BULLETIN OF MEDICAL PUBLICATIONS : W. B. SAUNDEUS, PHILADA. JUST PUBLISHED. A TEXT-BOOK OF MATERIA MEDICA, THERA- PEUTICS, AND PHARMACOLOGY. By GEORGE F. BUTLER, PH. G., M. D., Professor of Materia Medica and of Clinical Medicine in the College of Physicians and Surgeons, Chicago ; Professor of Materia Medica and Therapeutics, North- western University, Woman's Medical School, etc. 8vo, 858 pages. Illustrated. Prices: Cloth, $4.00 net; Sheep or Half- Morocco, $5.00 net. A clear, concise, and practical text-book, adapted for permanent reference no less than for the requirements of the class-room. The arrangement is believed to be at once the most philosophical and rational, as well as that best calculated to engage the interest of those to whom the academic study of the subject is wont to offer no little perplexity. Special attention has been given to the Pharmaceutical section, which is exceptionally lucid and complete. ESSENTIALS OF PHYSICAL DIAGNOSIS OF THE CHEST. By ARTHUR M. CORWIN, A. M., M. D., Demonstrator of Physical Diagnosis in the Rush Medical College, Chicago; Attending Physician to the Central Free Dispensary, Department of Rhinology, Laryngology, and Diseases of the Chest. 200 pages. Illustrated. Cloth, flexible covers. Price, $1.25 net. ARCHIVES OF CLINICAL SKIAGRAPHY. By SYDNEY ROWLAND, B. A., Camb., Late Scholar of Downing College, Cambridge, and Shuter Scholar of St. Bartholomew's Hospital, London; Special Commissioner to "British Medical Journal" for the Investigation of the Applications of the New Photography to Medicine and Surgery. A series of collotype illustrations, with descriptive text, illustrating the applications of the New Photography to Medicine and Surgery. Price, per Part, $1.00. Parts I. and II. now ready. FORTHCOMING PUBLICATIONS. SURGICAL DIAGNOSIS AND TREATMENT. By J. W. MACDONALD, M. D., Graduate of Medicine of the University of Edinburgh ; Licentiate of the Royal College of Surgeons of Edinburgh ; Professor of the Practice of Surgery and of Clinical Surgery in Minneapolis College of Physicians and Surgeons; Surgeon to Wisconsin Central Railway, etc. TEXT-BOOK OF EMBRYOLOGY. By JOHN C. HEISLER, M. D., Prosector to the Professor of Anatomy, Medical Depart- ment of the University of Pennsylvania. , BULLETIN OF MEDICAL PUBLICATIONS : W. B. SAUNDERS, PHILADA. Now Ready, Volume for 1896. SAUNDERS' American Year-Book of Medicine and Surgery COLLECTED AND ARRANGED BY EMINENT AMERICAN SPECIALISTS AND TEACHERS, UNDER THE EDITORIAL CHARGE OF GEORGE M. GOULD, M. D. NOTWITHSTANDING the rapid multiplication of medical and surgical works, still these publications fail to meet fully the requirements of the general physician, inasmuch as -he feels the need of something more than mere text-books of well- known principles of medical science. Mr. Saunders has long been impressed with this fact, which is confirmed by the unanimity of expression from the profession at large, as indicated by advices from his large corps of canvassers. This deficiency would best be met by current journalistic literature, but most practitioners have scant access to this almost unlimited source of informa- tion, and the busy practiser has but little time to search out in periodicals the many interesting cases, whose study would doubtless be of inestimable value in his practice. Therefore, a work which places before the physician in convenient form an epitomization of this literature by persons competent to pronounce upon The Value of a Discovery or of a Method of Treatment cannot but command his highest appreciation. It is this critical and judicial function that will be assumed by the Editorial staff of the " American Year-Book of Medicine and Surgery." It is the special purpose of the Editor, whose experience peculiarly qualifies him for the preparation of this work, not only to review the contributions to American journals, but also the methods and discoveries reported in the leading medical journals of Europe, thus enlarging the survey and making the work characteristically international. These reviews will not simply be a series of undigested abstracts indiscriminately run together, nor will they be retro- spective of "news" one or two years old, but the treatment presented will be synthetic and dogmatic, and will include only what is new. Moreover, through expert condensation by experienced writers, these discussions will be COMPRISED IN A SINGLE VOLUME OP ABOUT 1200 PAGES. The work will be replete with original and selected illustrations skilfully reproduced, for the most part, in Mr. Saunders' own studios established for the purpose, thus ensuring accuracy in delineation, affording efficient aids to a right comprehension of the text, and adding to the attractiveness of the volume. Prices: Cloth, $6.5O, net; Half -Morocco, $7.5O, net. W. B. SAUNDERS, Publisher, 925 Walnut Street, Philadelphia. 14 DAY USE RETURN TO DESK FROM WHICH BORROWED BIOLOGY LIBRARY TEL. NO. 642-2531 This book is due on the last date stamped below, or on the date to which renewed. Renewed books are subject to immediate recall. MAR1 -4 1973 APR- 2 1973 2 LD 21A-15m-2,'69 (J6057slO)476 — A-32 General Library University of California Berkeley 71763 M 6 THE UNIVERSITY OF CALIFORNIA LIBRARY