Se Ki ‘s ae : aN i a Let hallis ay rN NY ne ay le mek VEER AYE Vie a y my ae ee et ‘ a ' . f i Aas AAI AL A LAUER: iu AYath: Raed ! j toute Sees =i ee : CORNELL UNIVERSITY THE Flower Heterinary Library FOUNDED BY ROSWELL P. FLOWER for the use of the N. Y. STATE VETERINARY COLLEGE 1897 This Volume is the Gift of Dr. V. A. Moore. 5577 LIBRARY iw MWllows Cornell University Library The original of this book is in the Cornell University Library. There are no known copyright restrictions in the United States on the use of the text. http://www. archive.org/details/cu31924057010575 THE PATHOLOGY AND DIFFERENTIAL DIAGNOSIS OF INFECTIOUS DISEASES OF ANIMALS Prepared for Students and Practitioners of Veterinary Medicine By VERANUS ALVA MOORE, B.S., M.D., V.M.D. Brofessor of Comparative Pathology, Bacteriology and Meat Inspection, New York State Veterinary College at Cornell University, and Dean of the College FOURTH EDITION REVISED AND ENLARGED WITH 120 ILLUSTRATIONS NEW YORK THE MACMILLAN COMPANY 1916 All rights reserved "me eames aes oe ian A. MOORE COPYRIGHT I906 BY VERANUS A. MOORE COPYRIGHT 1908 BY CARPENTER & CO. COPYRIGHT I916 BY THE MACMILLAN COMPANY New and Revised Edition Published September, 1916 SF 16! M an 1G ib PRESS OF W. «. HUMPHREY, GENEVA, N. Y. TO DANIEL ELMER SALMON LARGELY THROUGH WHOSE LABORS THERE WAS ESTABLISHED IN THE UNITED STATES DEPARTMENT OF AGRICULTURE THE BUREAU OF ANIMAL INDUSTRY WHICH HAS MADE POSSIBLE EXTENSIVE INVESTIGA- TIONS INTO THE NATURE OF EPIZOOTIC DISEASES IN AMERICA AND WHO FOR TWENTY-ONE YEARS DIRECTED THESE INVESTIGATIONS THIS VOLUME IS DEDICATED. PREFACE The purpose of the Fourth Edition of this book is to provide students and practitioners with a text that will give them information on the etiology and morbid anatomy of the specific infectious diseases of animals and the methods available for their diagnosis. It is possible, within the limits of a workable text book, to include but a small part of the available knowledge on each of these diseases. To supplement the necessarily brief account, a list of the more important publications is appended to the description of each. It is believed that these references will give the key to the literature thereby making it possible for the student to familiarize himself with the present knowledge of the subject. The sanitary significance and the economic importance of the infectious diseases of animals are calling for a better understanding of their nature and more efficient methods for their control. These will be attained only through a more definite and specific knowledge of the etiology and morbid anatomy of each of these maladies. This edition has been carefully revised, much of it rewritten and numerous additions made. It has been kept, however, within the limits of a text book. Two appendices have been added, one on the requirements for interstate shipment of live stock and the other on the Federal regulations for the veterinary inspection of meat. These may be of much assistance to veterinarians. The diseases not indigenous to, or imported into, this country have been accorded much less space than those existing here. The desire is to emphasize the nature of the diseases our veterinarians are liable to encounter and, at the same time, give the characteristics of the others. The same plan of presenting the subject and of grouping the diseases according to their etiology that was followed in the previous editions has been retained in this. J desire to express my appreciation of the kind reception accorded the third edition, and for the helpful suggestions received from its readers. It is hoped that this edition will be still more useful to the student and practitioner. My thanks are especially due to Dr. C. P. Fitch for making several of the photographs particularly those of glanders, reproduced in the the text. V. A.M. v PREFACE .TO THE FIRST EDITION The literature on infection and the etiology and morbid anatomy of infectious diseases of animals is exceedingly rich in the results of new discoveries and important investigations. However, students just beginning this study and following a prescribed curriculum have not the time nor are they prepared to read with profit the detailed records of original research. Such publications seem to be better adapted for those doing advanced or graduate work. Further- more, many of these publications are out of print and are only avail- able for consultation. For these reasons it is believed that a volume containing the rudiments of the subject will be of use to the student and an aid to the teacher. It is also believed that such a work will be of assistance to practitioners. In preparing this volume the aim has been to bring together in a concise form the fundamental facts in the pathology of the more common infectious diseases of animals, especially those existing in the United States, with which sanitarians and the practitioners of com- parative medicine must contend. To this end the current literature, the reports of the investigations made at various institutions and experiment stations, as well as the standard works on comparative pathology have been freely drawn upon, to all of which full acknowl- edgment is hereby made. In order to bring into consideration the clinical value of a knowl- edge of morbid anatomy a few of the symptoms or antemortem manifestations have been included. It is hoped that this correlation of symptoms and lesions will stimulate a deeper interest in the study of comparative pathology and thus render it of more permanent and practical value for those entering into the practice of veterinary medicine. In selecting the subject matter care has been taken to avoid, as far as possible, the introduction of results concerning which there are controversies. It has seemed best to deal with those facts about which at the present time there is little or no doubt. After the discussion of each disease a few references to the literature are ap- pended. These are intended simply to bring the attention of the student to afew publications respecting the cause and morbid anat- omy, considered in the light of modern etiology, of the disease in question and to a few articles containing the results of original research. vil vill PREFACE In order not to complicate or unnecessarily expand this text, a knowledge of general pathology and the principles of bacteriology has been taken for granted. The difficulties involved in the preparation of such a text are both numerous and obvious. The indication of errors or omissions with any other criticisms that would tend to better the volume and increase its efficiency for the student will be thankfully received. V. A.M. TABLE OF CONTENTS PAGE List: of Wustrations 235. p23 6a... qculdwasarsn ceeded Midian ean gee enndasaaay xiil List of reference books. 2.20.06... 0000000 c ccc e cece eee cecaetvtenteesnees xvi CHAPTER I Erro.oey, Inrection ano Speciric Inrectious D1srasEs Etiology wipes. veges wie qaienace Os oceis a8 diadlaiead aon hed oie Ace marnmmcempnenek ook 1 Taf Chi Oris aise: es -eisionies sti cemte vie avons 8 1 deduces cats a Simca eh atmenennseteeeld a hoor 4 Woutidinfection) 23.01.05 nade vasnaabardoues axiaaen pea ee ORBeGlea eens sae 4 A specific infectious disease... 20.00.26 e cence cece eeeees 7 Dissemination of specific infectious diseases... .......0.0.0. 0.0 c cece ceeeeeeees 10 Diagnosis of specific infectious diseases... 0... 0.26. ccc cece cence 12 Classification or grouping of the infectious diseases...........0.....00.0.0 eee 16 BotryOmycosis! .22.5.2. pdnasaw Gaatedha 4 pita edi ander cupotaeanianan inna aes 19 Omphalophlebitis or navel ill... 20.0020 cece cece ences 20 White scours or diarrhea in calves... 20.2.2... cc cee eee nes Qi Infectious suppurative cellulitis of the limbs..............0..0000 00000 c cece 24 Fistulous withers and poll evil... 0000... 0000 cece cece eee eee eeeveeeeeeees 25 Infectious mastitis... 04 ccu ped eee cas isin ems ben peebsbeunbuwamanniaabeds 25 Contagious algalactian. 3 cuir seach beniieia BAe RA Aa daar PARR ES Oto 28 CHAPTER II Diseases Causep By Bacrerta—GeEnvus STREPTococcus General discussion of streptococci.......... 00 cece cece e eens 30 Classification of streptococci... 2.0... 6... e ens 31 Distribution of streptococci in nature......... 00... eee eee 31 Pathogenesis of streptococci... ........ 6c ene eee ete eee 32 Strangles ccf astuswan ties net Cage ioe pas Meta iaan oer hae ileneeaNes 33 Apoplectiform septicemia in chickens. .........00000 000000 c eee cece eens 38 Streptococcic:mastitiss «caress sagee ker yse reas sewed sey eee ered ie memmiA 40 CHAPTER III Diseases Causep BY Bactrerta—GeEnvus Micrococcus General discussion of the genus micrococcuS..............-20 000 c eee e eee 42 ST alcOS18 Ss pte to eh lalate caraeaqes anche pa sabanee yee wah epa ar aR aan Gens aatads 42 CHAPTER IV Disrases Causep By Bacrerra—Genus Bacterium General discussion of the genus bacterium. ............. 60.50. e cece eee eee 47 Swine plagues :cccich a aarewaccn ad oocaes aamnaihal aihy cisstnerhstudeanucorinas Hae 8 AOE RES 48 Hemorrhagic septicemia in cattle... 2.0.0... 2 octets 59 Fowl] cholera.......... Stile ages el arae achat ohh odia aE RT leat aE OE 69 Goose septicemia ... 2.20.00... cece eee ce ee terete ete n eee n enn eeeee 74 Fowl typhoid 3 zc scy-c00-b bdnciusio rons 2 65 eae EY AEE EE a ea eAbIRaE Ss 17 x TABLE OF CONTENTS , PAGE Swine erysipelas... 0.0.0.0... cece cece cece ce eee e bebe cee eee eeteeieeeeees 84 ADthTaxy diencscd yeaa ene sgn breeu gun Maulana ect Meng onemunmndg seaeies 89 Glanders. 3. :.54 secant ese 58454845 COSOROGEN Do dearest da bebe tum pa 109 Tubereulosis. seco ausisigcisescatoaiy eae ehaw aeetuscnad oe BS HESS ELA ARR eS 146 JOLIE'S ISCAS, a iis inanaen da Ric Gulad an nGlenme ADE ha DERG ea moneio eee RES 191 Infectious abortion in cattle... 0.06... c eee teen eee eens 198 Ovine caseous lymph adenitis (pseudo-tuberculosis in sheep)...............045 209 Astheniainfowlsand pigeons) .c0.2.2005 songsbald , 498 Corn stalk disease in cattle... 2.00... ccc nee eee eee n ete ne naee 500 CHAPTER XIV Immunity anp Protective INocuLATION Natural immunity 555 2340455 wanton cae sca ese ee Ramee esaiassaaasacaiena te Greeaiadesuacandn tines BLOOD COUNTS OF APPARENTLY HEALTHY FOWLS Fowl. White Corpuscles. Red Corpuscles. NOs dhcasuseweeens poereMeaneets ieee meets S 24,000 2,980,000 per cmm. ING iy Rezaiciic hiticaian cd crt Sadecaeytodd Sushaiin tra sabdstansesenanteaen wt 26,300 2,987,000 “ <“ NIN ete erate Bs ARS AEA aa a nh Aiea 36,000 8,115,000 “ “ IN Gis iB sce sesrcsnctrssteauietesrace sane ee sbcacacing ent se oper eseme a ai 52,000 3,980,000 “ “ ING. AG 6 serracceny cakeeummsummery aeaeloenemmee ye 61,000 3,920,000 “ “ NO. ll sse¢ cane w ss daceaneeneseaearauee ane 30,000 2,380,000 “ “ NGOs 1B) ia scidieraae Saree mencoas tat Oteoatamatese 24,000 2,620,000 “ “ Diagnosis. Fowl cholera is diagnosed by the symptoms, lesions and bacteriological examination. The specific cause is not difficult to isolate. Fowl cholera is to be differentiated from: A number of dietary disorders which cause the death of a large number of fowls. Such cases are often thought to be chicken cholera and so reported by the owners. A diagnosis is to be made from the bacteriological findings. It is to be differentiated from fowl typhoid. There are a number of resemblances in the clinical history of the two diseases. There are, however, marked differences in both the morbid anatomy and etiology For a comparison see fowl typhoid. Prevention. Pasteur introduced a preventive inoculation or vaccine for this disease. Kitt found that the eggs of fowls unknown to this disease possessed a substance somewhat similar to antitoxin. He immunized fowls by injecting them simultaneously with from four to eight cubic centimeters of the whites of such eggs. More recently he has obtained a horse serum that promises to be of im- munizing value. Jensen obtained good results in immunizing geese against this disease by the use of serums. There does not appear, however, to be a reliable immunizing agent. Hadley has used sub- cutaneous injections of diluted carbolic acid. 74 GOOSE SEPTICEMIA Good sanitary conditions, isolation of the well from the sick fowls and thorough disinfection seem to be the most satisfactory procedure. It is important not to introduce the disease with newly purchased fowls, or to expose healthy ones to the disease either at or in trans- portation to various poultry exhibits. Control. Fowl! cholera is a reportable disease in Germany, Austria and Hungary. In those countries the infected premises are quaran- tined against traffic in fowls. The infected places should be thor- oughly disinfected. Care should be taken not to introduce infected fowls into healthy flocks. The fowls that appear to be sound but which are in either the period of incubation or have recovered from the disease are very liable to spread the infection. J REFERENCES 1. Haviey. Fowl cholera and methods of combatingit. Bulletin 144, R. I. Agr. Exp. Station, 1910. 2. Hicerns. Notes upon an epidemic of fowl cholera and upon the comparative production of acid by allied bacteria. Jour. of Experimental Medicine, Vol. III (1898), p. 651. 3. Kirr. Die pone alpine gegen Gefliigelcholera. Monatshefte fiir praktische Tierheilk., Vol. XVI (1904), S 4. Prrroncito. Ueber al epizootische Typhoid der Hithner. Arch. fiir wiss. u. prackt. Thierheilk., 1879. 5. Pasteur. De l’attenuation du virus du cholera des poules. Comptes rendus des Seances del Academie des Sciences, Vol. XCI (1880), p. 673. 6. Pasteur. Sur les maladies virulentes, et en particulier sur la maladie appelée vulgairement choléra des poules. Ibid. Vol. XC (1880), p. 239. 7. Satmon. Annual Reports of the U. S. Commissioner of Agriculture, 1880-82. 8. Satmon. The diseases of poultry. Washington, D. C. 1889, p. 232. V9. Warp. Fowl cholera. Bulletin No. 156. College of Agric., Calif. Agric. Exp. Station, 1904. GOOSE SEPTICEMIA Characterization. The disease is an acute septicemia causing the death of the infected goose in a few hours after there are evidences of sickness. History. In 1902, Curtice described this disease as causing con- siderable loss in Rhode Island. The following note by T. Smith, dated October 31, 1900, quoted by Curtice, is significant in explaining the condition under which the disease appeared. “Geese born in April and May and collected during the summer and fall for fattening, kept in open yards, crowded together but able to move about; about 500 in a pen. Fed on a mixture of corn meal and meat and beef scraps. Epidemic began in mid- summer. Deaths up to twenty a day (one workman says sixty one day); about 3,000 lost to date.” GOOSE SEPTICEMIA 75 Etiology. The cause of this disease is a bacterium belonging to the septicemia hemorrhagica group. It is stated to have “the characters of the fowl cholera type.” It killed rabbits when they were inocu- lated with 0.2 ce. of a bouillon culture. Symptoms. The symptoms are indefinite. In the outbreak described the geese were often found dead. The description of the disease by Curtice is appended. “Few symptoms of disease were seen, those noted pertaining mainly to the death struggles. Very few that died were noticed to be sick more than an hour or two before death, and, as the experi- mental investigation demonstrated, the disease could not have lasted, in the majority of the geese, more than thirty-six hours. An uncer- tain gait, a burrowing of the head in the dirt, twisting it around, or actions indicating spasms of the throat, were the earliest symptoms. Some geese were observed to die within five minutes or after the first seizure.” There are few chronic cases and recoveries are not re- corded. Some show no other symptom than being slower in action, and separating themselves somewhat from the flock. However, this sign is quite important when the wild nature and gregarious habit of the goose are taken into account. Morbid anatomy. The tissue changes, as given by Curtice, are as follows: “There was considerable mucus in the throat and mouth, and a very tenacious mucus in the nose. The veins of the head were usually congested, as though the animal had died of asphyxia. This, together with spasm of the throat, indicates a spasmodic closure of the glottis. The digestive tract was found to be full of food in nearly all stages of digestion. In some cases the catarrhal products of the intestines contained petechize. Sometimes these points were collected in more or less extensive patches. Perhaps more than half of the livers showed yellow spots of from a pin point to a pin head in size. These discolorations were found on section to extend into the sub- stance of the liver, and were evidently dead tissue, or necroses. In one example the heart disclosed severe inflammation, both epicarditis and pericarditis being present. In one case the lungs were affected. In all, fifteen cases were examined, and from these this composite description of the post mortem appearances is drawn.” Hemorrhages on the serous membranes and punctate necroses in the liver seem to be quite characteristic lesions. 76 GOOSE SEPTICEMIA Post mortem notes.—These are a few taken from Curtice’s publication: “Goose No. 1. Died last night; quite fat. Right lung, ventral portion quite firm, whitish. Some flocculi of exudate in peritoneal cavity. Liver shows numerous point- like necrotic foci. Blood thick, blackish and tarry. Mucus glassy on dusky mucosze of nose and throat. “Goose No. 2. Died last night. Somewhat thinner than No. 1. Ecchymoses on fat in abdomen and gizzard and on heart muscle; necrosis in liver. Blood thick, tarry. Mucus in nasal passages. “Gander No. 7. Died last night; now cold. No well marked hemorrhagic lesion in pleuroperitoneal cavity. Whitish points in liver. Hemorrhagic or extremely hyperemic condition of duodenum. Jejunum, or second coil of intestine, filled with a glairy mucous fluid in which are suspended shreds and patches of food (?). Few if any necroses in liver.” Diagnosis. Goose septicemia is to be diagnosed from the bac- teriological examination. It is caused by a pasteurella which resem- bles that of fowlcholera. A diagnosis, therefore, is made positive by finding this organism in the tissue of the sick and dead geese. It is to be differentiated from other forms of infection from which geese may suffer. M’Fadyean has described a disease under this title causing the death of many geese in which he found the blood swarming with bacteria suggesting Bact. septicemiae hemorrhagicae but morphologi- cally different. He could not induce it to grow on any of several media in cultures under both aérobic and anaérobic conditions. It appears that this is a different disease from that described by Curtice. Prevention. The procedure that can be suggested at present is isolation of the well from the sick, repeating the separations as often as new cases appear. The infected pens should be thoroughly dis- infected before being reoccupied. REFERENCES 1. Curticr. Goose septicemia. Bulletin No. 86, R. I. Agr. Exp. Station, 1902. 2, M’Fapyran. A remarkable outbreak of goose septicemia. Jour. Compar. Path. and Therap., Vol. XV (1902), p. 162. Fowls and poultry of all kinds are subject to infections that are interesting in their nature and often fatal in their results. There is a large literature on the subject. The general statement may be made that all poultry seem to be susceptible to the Pasteurella. FOWL TYPHOID 17 FOWL TYPHOID Characterization. A specific disease of fowls caused by Bacterium sanguinarium. It is not known whether or not other species of domesticated birds are susceptible. History. This disease was briefly described by Moore in 1895. At that time it had been studied in but a few fowls and these the last to die in their respective flocks. In the following year other fowls. were examined very carefully from two outbreaks of the disease and it is upon the data obtained in this investigation together with those procured from many produced cases that the description of the disease is based. It was described as an infectious leukemia. Further investigation, however, has shown that the excess of white corpuscles. was due to a leucocytosis brought about by the infecting organism and that the disease is not a leukemia. It was found by Smith in 1894, on Block Island, R. I. In 1898, Dawson found it to be the cause of very serious losses among poultry near Baltimore, Md. In all of the outbreaks studied, the owners of the fowls first reported the disease as chicken cholera. In 1902, Curtice investigated an outbreak in Rhode Island. Geographical distribution. It was first studied in fowls taken from an outbreak in Virginia. Since then, it has been identified in Maryland, the District of Columbia, and the State of Rhode Island. There is good evidence in the numerous reports of destructive fowl diseases to believe that it is quite widespread in the United States. Etiology. Moore isolated and described a pathogenic bacterium which he designated Bacterium sanguinarium. With this organism the disease has been produced in healthy fowls both by feeding cultures and by intravenous injections. Its etiological relation to the disease is, therefore, quite clearly established. It is possible that certain accompanying conditions may be necessary in conjunction with the organism to cause the disease to spread rapidly in a flock. Experimentally it did not spread from diseased (inoculated or fed) to healthy fowls when kept in the same yard. Symptoms. From the statement of the owners of the diseased fowls in the different outbreaks and from the appearance of those in which the disease was artificially produced, little can be positively stated concerning the early symptoms. There is a pronounced anemic condition of the mucosa of the head. An examination of the 78 FOWL TYPHOID blood shows a marked diminution in the number of red corpuscles and an increase in the number of white. ones. In the disease pro- duced artificially by feeding cultures of the specific organism there is, in most cases, a marked drowsiness and general debility mani- fested from one to four days before death occurs. The period during. which the prostration continues varies from a few hours to two days. The mucous membranes and skin about the head become pale. There is an elevation of from 1 to 4 degrees in temperature. The fever is of a continuous type, as shown in the appended temperature chart of two fowls in which the disease was produced artificially. Although the course of the disease in different fowls is usually constant, there are many variations. The time required for fatal results is from three to fifteen days, but ordinarily death occurs in about eight days after feeding the cultures. The rise in temperature can be detected about the third day and external symptoms about the fifth or sixth, occasionally not until a few hours before death. The symptoms observed in the cases produced by feeding correspond with those described by the owners of affected flocks. As indicated in the inoculation experiments, the symptoms follow- ing the intravenous injection of the virus were, as would be expected, considerably modified from those fowls which contracted the disease by the ingestion of cultures of the specific bacterium. FAHR. £ 6 7 c) 7 10 4 12 73 3" “2 mw A 7 nla es Ddad, a 1 <=" IN_ > “lt _pdea 0 He WK ze iN ee : we 7 109 at ; wa ‘ v = -~ “Se L— 7 J - \] a Us 107 Za 1 > a sade | roel __ Fic. 7. TemMprerRaATURE CHART OF TWO FATAL CASES ARTIFICIALLY PRODUCED IN FOWLS. Morbid anatomy. The only constant lesions found in the fowls which contract the disease naturally, as well as in those fed upon the virus, are in the liver and blood. The liver is somewhat enlarged and dark colored. A close inspection shows the surface to be sprinkled with minute grayish areas. The microscopic examination shows the blood spaces to be distended. The hepatic cells often stain very FOWL TYPHOID 79 feebly. Not infrequently the cells are isolated and their outlines indistinct. Occasionally foci are observed in which the liver cells appear to be dead and the intervening spaces infiltrated with round cells. The changes in the hepatic tissues are presumably secondary to the engorgement of the organ with blood. The rareness with which the intestinal tract is affected in both the natural and artificially produced cases is exceedingly interesting and important for the differential diagnosis. There is in most cases a hyperemia of the mucous membrane of the colon, but this condition is not uncommon in the healthy individual. The kidneys are gener- ally but not uniformly pale. They are streaked with reddish lines, due to the injection of blood vessels. In section the tubular epi- thelium appears to be normal. The kidneys seem to be, from the number of bacteria in the cover-glass preparations, especially favor- able for the localization of the specific organism. The spleen is rarely discolored or engorged with blood. The lymphatic glands were not appreciably enlarged in any individual examined. The lungs except in chronic cases are normal. The brain and spinal cord are unaffected. The heart muscle is usually pale and sprinkled with grayish points due to cell infiltration and necrosis. These lesions are so common that it seems safe to consider them characteristic manifestations. Death usually occurs in systole, the auricles containing very thin, unclotted blood. The most important alterations are found in the blood. These consist, in the progress of the disease, of the gradual disappearance of the red corpuscles and increase in the number of white ones, as determined by blood counts made daily or every other day, from the time of inoculation, or of feeding the virus, until the day of death. The diminution in the number of red corpuscles and the increase in the number of white ones are illustrated in the blood count of two eases of artificially produced disease. In carefully heated cover-glass preparations of healthy fowl’s blood stained with methylene-blue and eosin, the nuclei are colored a deep blue, and the cellular protoplasm surrounding the nucleus is stained by the eosin. In similar preparations made from the blood of the affected fowls there are a greater or less number of cells which do not take the eosin stain. These were called spindle cells by Van Recklinghausen, blood plates by Bizzozero, and hematoblasts by Hayem. More recently Dekhuyzen has called them thrombocytes. 80 FOWL TYPHOID In these the portion of the cell body surrounding the nucleus remains unstained or becomes slightly tinted with blue. Occasionally they contain one or more vacuoles, and the margin is frequently broken. The apparent dissolving away of the red corpuscles has been fre- = Fic. 8. Bioop rRoM A WELL ADVANCED CASE OF FOWL TYPHOID SHOWING RED CORPUSCLES, BLOOD PLATES AND IN- CREASE IN THE NUMBER OF LEUCOCYTES. quently observed and corpuscles- showing the intermediate stages are readily detected in carefully prepared specimens. These must be differentiated from the blood plates. The cause of the destruction of the red corpuscles is not satis- factorily explained. In his re- port on fowl cholera, Salmon illustrates leucocytes surrounding the red corpuscles, but the marked diminution of the red cells was not determined. He _ speaks, however, of the pale color of the blood. In fresh preparations of the blood, portions of red cells may be seen within the leucocy- tes, those containing spindle- shaped granules. The determi- nation of the extent of this mode of destruction of the red corpuscles necessitates further investigation. TABLE SHOWING CHANGES IN THE NUMBER OF CORPUSCLES Fowl inoculated in the wing vein February 6 Number of Number of Date |Temperature ted cor- white cor- Remarks (F°.) puscles puscles per c. mm. per c. mm, Feb. 6 107.4 3,744,444 21,222 Well vi 109 3,417,391 26,087 | Apparently well. 8 108.2 2,784,700 55,000 Do. 9 108.4 2,807,692 76,925 Do. 11 107.4 3,481,818 90,909 Feathers ruffled; refuses food. 13 110.2 | | 2,133,333 100,000 | Very quiet; comb pale. 14 108 2,530,000 140,000 | Fowl died later in the day. FOWL TYPHOID 81 Fow! fed culture March 26 7 : saree of Meet of Date emperature red cor- white cor- (F°) puscles puscles Remarks perc, mm, per c, mm. Mar. 26 106.2 3,535,000 18,940 Well. 28 110 2,430,000 70,000 Fowl eats very little. Apr. 2 110.6 1,684,210 80,000 Blood very pale; fowl weak; re- fuses food. 8 106 1,745,000 245,000 Very weak; many red corpuscles attacked by leucocytes. Ae le ett ia nace ath ak wink cae e-card eran ean eect Found dead. In fresh preparations of the blood of affected fowls examined in Toisson’s fluid, red corpuscles which take the violet stain more or less intensely throughout are frequently observed. In the blood of poultry two distinct classes of white corpuscles are conspicuous. The first, which predominates in numbers, contains nuclei with from one to four lobes, and the cytoplasm is sprinkled with a variable number of round, elongated, or spider-shaped bodies. In the fresh condition they are highly refractory. They stain with eosin, and if the preparations are heated sufficiently they will retain certain of the aniline dyes. The other class consists of round or nearly round cells which takes the blue stain feebly. Usually it is difficult to detect the nucleus, although it is occasionally distinct. Between these two types there are many varieties. The leucocytes containing the spindle-shaped bodies appear to be the phagocytes, as they were the only ones which were observed to engulf the red corpuscles. Bacteria have not been demonstrated in these cells, although their presence has, in several cases, been suspected. From the appearances observed in the red blood corpuscles it seems highly probable that phagocytosis plays a comparatively large part in their destruction. Another hypothesis is also suggested, namely, that a toxin produced during the multiplication of the specific organism has this effect on the red corpuscles. In the fresh preparations we can observe the phagocytes attacking the red cells. In the stained ones mutilated red corpuscles and free nuclei are present. The hypothesis is suggested that the leucocytes partially digest certain of the red corpuscles in their attack upon them. Whether these changes are entirely attributable to the phagocytes is an open question. In the blood from healthy fowls it is comparatively rare to see one of the white corpuscles engulfing ared one. As the disease progresses, 82 FOWL TYPHOID however, this warfare becomes very conspicuous, owing perhaps to the increased number of the colorless cells. Up to the present the study of these corpuscles has not been extended beyond the observa- tion of the general appearance of these structures, and no attemptis made to explain the apparently marvelous increase in the number of the leucocytes. It is an interesting and as yet unexplained fact that the increase in the corpuscles is apparently restricted to those con- taining the spindle-shaped bodies. Diagnosis. Fowl typhoid is diagnosed by the symptoms, lesions and the finding of Bact. sanguinarium in the organs. It is to be differentiated from intestinal disturbances, especially diarrhea and fowl cholera. A comparison of the important changes in the morbid anatomy in fowl] cholera, as described by European writers, and in the disease under consideration, can be made from the appended columns, in which their more characteristic lesions are contrasted: LESIONS IN FOWL CHOLERA LESIONS IN FOWL TYPHOID 1. Duration of the disease from a few 1. Duration of the disease from a few hours to several days. hours to several days. 2. Elevation of temperature. 2. Elevation of temperature. 8. Diarrhea. 3. Diarrhea not common. 4. Intestines deeply reddened. 4. Intestines pale. 5. Intestinal contents liquid, muco- 5. Intestinal contents normal in con- purulent, or blood stained. sistency. 6. Heart dotted with ecchymoses. 6. Heart usually pale and dotted with grayish points. due to cell infiltra- tion. 7. Lungs affected, hyperemic or pneu- 7. Lungs normal, excepting in modified monic. cases. 8. Specific organisms appear in large 8. Specific organisms comparatively numbers in the blood and organs. few in the blood and organs. 9. Blood pale (cause not determined). 9. Blood pale, marked diminution in the number of red corpuscles. Attention should be called to the fact that as yet there seems not to have been a careful study of the condition of the blood in fowl cholera. Salmon observed many changes which may have been similar to or identical with those herein recorded. Ward found an increase in the number of white corpuscles and in some cases a decrease in the number of red ones in cases of fowl cholera. FOWL TYPHOID 83 The difference between the specific organisms of these two diseases can readily be appreciated by a comparison of the more diagnostic properties of each; they are arranged in parallel columns, as follows: BACTERIUM OF FOWL CHOLERA BACTERIUM SANGUINARIUM Bacterium short, with oval ends. 1. Bacterium short, with ends oval or more pointed. It usually appears singly in tissues. 2. It usually appears in pairs united end to end or in clumps in tissues. Ordinarily it exhibits a polar stain 3. It gives a light center, with uni- (from tissue). formly stained periphery (from tissue). Rarely a polar stain is observable. Grows feebly or not at all on gelatin. 4. Decided growth on alkaline gelatin. It does not change milk. 5. Saponifies milk.. Resists drying from one to three 6. Resists drying from eight to twelve days. days. Kills rabbits inoculated subcutane- 7. Kills rabbits inoculated intraven- ously in from eighteen to twenty- ously in from three to five days. four hours. Rabbits inoculated subcutane- : ously remain well or die in from six to ten days. It kills fowls when injected sub- 8. It does not kill fowls when injected cutaneously in small quantities. subcutaneously in small quanti- ties. While there are many similarities in the symptomatology of these two diseases, there are pronounced differences in the morbid anatomy and in the specific microérganisms. These facts render positive differentiation dependent upon a careful bacteriological and pathologi- cal examination. In fowl cholera the course of the disease is more rapid than in fowl typhoid. Prevention. Prompt isolation of the well from the sick fowls and thorough disinfection of the houses and yards. In reference to preventing its introduction, Curtice makes the following observation: “Inasmuch as one possible method of introducing the disease is through purchases, it will always be necessary for purchasers to inquire into the history of the flock from which additions are to be made, and especially to examine into the condition of the fowls. It is better in any case to keep new purchases by themselves for some weeks or until it is apparent that they are healthy.” 84 SWINE ERYSIPELAS REFERENCES 1. Curticr. Fowl typhoid. Bulletin 87. Agr. Exp. Station of the R. I. College of Agric. and Mech. Arts, 1902. 2. Moorr. A study of a bacillus obtained from three outbreaks of fowl cholera. Bulletin No. 8, U. 8. Bureau of Animal Industry, 1895. 3. Moore. Infectious leukemiain fowls—A bacterial disease frequently mistaken for fowl cholera. Annual Report of the Bureau of Animal Industry, 1895-96. SWINE ERYSIPELAS Synonyms. Red fever of swine; rouget du porc; Rotlauf. Characterization. This disease, peculiar to swine, is determined by a rise of temperature, cerebral disturbances and pronounced reddening of areas of the skin. It is a disease of adult life. It is stated that pigs are rarely attacked under three months or over three years of age. Lydtin and Schottelius found some differences in the degree of susceptibility of certain breeds of swine. The common country pig was least susceptible. History. This disease has been known in Europe for many years. It was not distinguished from other infections until studied by Pasteur and Thuillier. Smith found a bacterium in rabbits inocu- lated with the organs of pigs that had died of an undetermined disease in Minnesota, which was either the bacterium of swine erysipelas or of mouse septicemia. The latter organism had been recorded on two previous occasions from pigs in this country. Geographical distribution. Swine erysipelas occurs enzodtically and in epizodétics in most of the countries of Europe. It was formerly restricted in Bavaria to the districts along the Danube, and was entirely unknown in southern Bavaria (Kitt). It is stated that the disease tends to become enzodtic chiefly in valleys and low-lving plains which have slow-flowing streams and heavy, damp, clay soil; and that sandy and granite soils are comparatively free from it. It occurs chiefly during the months of July, August and September, although it appears sporadically during the winter months. It has not been described from the United States. Etiology. Loeffler and Schitz pointed out in 1885 that swine erysipelas was caused by a very slender bacterium (Bact. rhusio- pathiae) 1 to 2u. long and 0.3 to 0.4u. broad, straight or slightly curved, ends not rounded and in cultures often appearing in filaments. It is SWINE ERYSIPELAS 85 very closely related to the bacterium of mouse septicemia described by Koch in 1878. There is much uncertainty concerning the relation- ship of the bacterium of mouse septicemia to that of this disease. Smith has suggested that possibly the bacterium which has been found in this country may gain virulence sufficient to produce epi- zootics, if such is not already the case. It is exceedingly important that careful search be made for this organism in the outbreaks among swine where the nature of the disease is not clearly determined. House mice and pigeons are susceptible to the bacterium of swine erysipelas; guinea pigs and fowls are immune. The bacterium of swine erysipelas is to be differentiated from that of mouse septicemia. The period of incubation is stated to be at least three days. It is apparently longer than that in many cases. Symptoms. The disease usually begins suddenly and violently. The animal refuses food, makes efforts to vomit, has a rise of tempera- ture, manifests severe nervous disturbance, is very weak, torpid and indifferent to its surroundings. When approached it tries to hide itself under its bedding. The hind quarters become weak and paralyzed. Muscular spasms and grinding of the teeth are sometimes observed. At first there is constipation, the conjunctiva is of a dark red or brownish-red color, and the eyelids are sometimes swollen. Usually a day or two after the first symptoms develop, or, perhaps, from the first, reddish spots appear on the thin parts of the skin, such as the region of the navel, lower surface of the chest, perineum, inner surface of the thighs, ears and throat. These spots, which at first are bright red and about the size of a man’s hand, become, later on, dark red or purple, and soon unite into large, irregularly-shaped patches. As arule, they are neither painful to the touch nor promi- nent, but sometimes they show a slight inflammatory swelling. The skin of the red spots, especially of the ears, may suffer from an erup- tion of vesicles and may even slough. Gangrene of the skin sometimes occurs. The reddening of the skin may be very slight in severe cases, or may appear only immediately before, or even after death. Death takes place usually on the third or fourth day. In the very severe form, the animal may die in twenty-four hours, otherwise the disease requires a week or longer to run its course. Jensen considers that this disease, instead of being uniform in its clinical aspects, manifests itself in the following forms, which differ from each other by well-marked peculiarities. The forms recognized 86 SWINE ERYSIPELAS as varieties of this disease but more generally considered as distinct maladies and known by different names are as follows: True erysipelas. Swine urticaria. Erysipelas without redness of the skin. Diffuse necrotic erysipelas of the skin. Endocarditis of erysipelas. He also maintains that there may sometimes be transitional forms between the respective varieties which he enumerates. Different forms of epizodtic erysipelas have also been described by Cornevin, Hess and others. The duration of the disease varies from 1 to 10 days. In ‘types of moderate severity it runs from 3 to 4 weeks. The prognosis is unfavorable. There is from 20 to 80 per cent. mortality. Morbid anatomy. In the ordinary form of epizodtic erysipelas there is a septicemic condition without any well marked morbid changes of separate organs. In less acute cases the septicemia may give way to hemorrhagic and diphtheritic gastro-enteritis, consider- able swelling of the lymphatic system, hemorrhagic or parenchyma- tous nephritis, and hepatitis, acute swelling of the spleen and myositis. The hemorrhagic gastro-enteritis consists at first of excessive inflam- mation of the mucous membrane of the stomach in the region of the fundus. The mucosa shows a dark-red discoloration which is partly diffuse and partly in spots. The cells suffer from cloudy swelling and the mucous membrane is covered with a viscid layer of mucus. The intestinal mucous membrane is swollen, especially on the top of the folds and in the nieghborhood of Peyer’s patches. It is infiltrated with blood and sometimes shows superficial scabs. Less frequently, circumscribed parts of the mucosa of the cecum and the anterior parts of the colon suffer from a diphtheritic affection. Ay The solitary follicles and Peyer’s patches appear as prominently raised patches. Sometimes they are infiltrated with blood and sur- rounded by a reddish band. There is ulceration and cicatrization of: the solitary and agminated follicles. The mesenteric glands become more swollen than the other glands of the body, of a dark red color, and show softening. ‘The surface of fresh sections is dun-colored with interspersed dark-red areas. The paraglandular tissue is hyperemic and infiltrated with blood. SWINE ERYSIPELAS 87 The kidneys are enlarged, the cortex of a grayish-red and the medul- lary portion of a very dark-red color. Frequently catarrhal nephritis occurs as a complication. The acute swelling of the spleen arises in consequence of an acute hyperemia, with an increase of the cellular constituents of the pulp, in which case the organ is enlarged, but not softened as in anthrax. The pulp is of a purple color, moderately soft and free from hemor- rhages. There is cloudy swelling and enlargement of the liver. The surtace of sections has a grayish-brown color, and the acini are widened. The muscles are gray in color, soft, flaccid, watery, glistening and some- times they are sprinkled with hemorrhages. They give the general appearance of boiled flesh. The myocardium shows similar spotted changes, and punctiform hemorrhages beneath the endocardium. In the abdominal and thoracic cavities and pericardium, there may be found small quantities of an orange-colored, clear fluid, which may be mixed with a flaky coagulum. Many English veterinarians regard the occurrence of more or less luxuriant vegetations on the valves of the heart to be so common that it is to be considered almost diagnostic. It would appear from the literature that this endocarditis is not nearly so common in conti- nental Europe. The lungs remain unchanged, or at most exhibit a post-mortem edema. By microscopic examination, the specific bac- teria are found everywhere in the body, especially in the spleen and kidneys, and to a less extent in the blood. Diagnosis. Swine erysipelas is diagnosed from the symptoms, lesions and the isolation of its specific cause. The Ascoli thermo- precipitation test may be used. It is stated by Gsawizky to be strongly specific and Seibold reports favorably on its use. It is the only specific test for indicating this infection. Swine erysipelas is to be differentiated from: Hog cholera and swine plague. The frequent reddening of the skin in these diseases together with the modified lesions so frequently observed may cause confusion. The bacteriological examination will enable the positive diagnosis to be made. Anthrax, which is very rare in swine. Here, in addition to the bacteriological examination, specific tests for anthrax can be em- ployed (see anthrax). Erythemata due to various dietary causes. The significance of a deep reddening of the skin about the head, abdomen and thighs of 88 SWINE ERYSIPELAS pigs is not fully determined. It is clear, however, that such a condi- tion often occurs in the absence, so far as present knowledge goes, of a specific infection. It is frequently found in pigs suffering from diges- tive troubles, or poisoning from eating decomposed offal. Prevention. Swine infected with this disease readily transmit it to others. The organism is reported to remain for a considerable time in the pharynx and nostrils and also to have been found in healthy swine. It is stated also to exist in the soil in a saprophytic form. These facts render prevention difficult. The general precaution of removing the sick from the well and not placing healthy swine in infected yards for some months after the recovery or removal of sick ones should be observed. Pasteur’s preventive inoculation -was until recently the chief pro- phylactic means employed against epizodtic erysipelas. Metchnikoff found that the blood of immunized rabbits was antitoxic, and Lorenz maintains that the serum of swine that have recovered from swine erysipelas is also antitoxic, and will produce immunity in other ani- mals. The treatment introduced by Lorenz is to inject the immuniz- ing serum in the proportion of 1 cc. to every 10 kilograms of the body weight of the animal. Two days afterward 0.5 to 1.0 cc. of virulent culture is injected, and after twelve days the dose is doubled. The use of the immunizing serum is reported to be very successful. Specific biologic treatment. The serum prepared by Lorenz is reported to give excellent results in acute cases. REFERENCES 1. Bane. Ueber Rotlauf-Endocarditis bei Schweinen. Deutsche Zeitschr. f. Thiermed., Bd. XVIII (1891), S. 27. 2. Jmnszen. Die Aetiologie des Nesselfiebers und der diffusen Hautnekrose des Schweines. Deutsche Zeitschr. f. Thiermed., 1892, S. 278. 3. Lorrrier. Experimentelle Untersuchungen tiber Schweine-Rotlauf. Arbeiten aus d. Kaiserlichen Gesundhettsamte, Bd. 1 (1885), S. 46. 4. Lorenz. Die Schutzimpfung gegen Schweinerotlauf mit Anwendung eines aus Blutserum immunisirter Thiere hergestellten Impfstoffes. Deutsche Zeitschr. f. Thier- med., Bd. XX (1894), S. 1. 5. Lorenz. Die Veterindrpolizeiliche Behandlung des Schweinerothlaufes und die Schutzimpfung. Berliner thierarz. Wochen., 1897, S. 574. ; 6. Lorenz. Schutzimpfungen gegen den Rotlauf der Schweine. Ibid., 1897, S. 09. 7. Moors. Mouse septicemia bacilli in a pig’s spleen with some observations on their pathogenic properties. Jour. of Comp. Med. and Vet. Archives, Vol. XIII (1892), p. 333. 8. Pasteur et TuurtuEr. La vaccination du rouget des porcs 4 Paide du virus mortel attenué de cette maladie. Comp. rendus Acad. des Sciences, Vol. XCVII (1883), p. 1163. ANTHRAX 89 9. Saumon. An Examination of Pasteur’s Va@ine for Rouget. Annual Report U.S. Bureau of Animal Industry, 1885, p. 187. 10. Scnurz. Ueber den Rotlauf der Schweine und die Impfung mit demselben. Arbeit a. d. Kaiserlichen Gesundheistamte, Bd. I (1885), S. 56. 11. Sersotp. Beitrag zur Feststellung des Rotlaufs der Schweine mit Hilfe der Thermo-pracipitinreaktions nach Ascoli. Zeit. f. infek. d. Haust., Bd. XIII, S. 91. _12. SairH. Swine erysipelas or mouse septicemia bacilli from an outbreak of swine disease. Annual Rept. U. S. Bureau of Animal Industry, 1895-96, p, 166. ANTHRAX Synonyms. Splenic fever; splenic apoplexy; wool sorters’ disease; malignant pustule; anthracemia; mycosis intestinalis; charbon; Milzbrand. Characterization. Anthrax is an infectious disease occurring sporadically and in epizoétics in herbivora and omnivora and com- municable to nearly all warm-blooded animals, and to man. It is characterized by a high temperature, the presence in the diseased tissues or liquids of Bacterium anthracis, by an enlarged spleen, blood extravasations and by local gangrene. It usually occurs in the acute form. Nearly all species of animals suffer from anthrax. The herbivora and rodents are most susceptible. Horses and mules often suffer from it. M’Fadyean has reported outbreaks aggregating 54 cases, of which 49 were cattle, 4 horses and 1 pig. He states also that for a period of 5 years there had been reported 192 cases in horses and 3,390 in cattle. It is interesting to note that the Algerian race of sheep are immune. A satisfactory explanation for this striking exception has not been recorded. It has been stated that a single bacterium introduced into the subcutaneous connective tissue of a guinea pig or mouse is sufficient to kill it. Cats, tame and wild rabbits and hares are the next most susceptible species. It is stated that dogs, pigs and foxes are very slightly susceptible. Rats, fowls and pigeons are re- ported to beimmune. Fish and amphibia are rarely attacked. History. Anthrax is among the oldest of the known infectious diseases of animals. Descriptions of epidemics and epizodtics of this disease are given by Homer, Plutarch, Livy and other writers before the Christian Era. The Arab physicians designated it as “Persian Fire.” Extensive outbreaks are mentioned in the literature of the fifteenth, sixteenth, seventeenth, eighteenth and nineteenth centuries. Chabert pointed out in 1780 that the various kinds or forms of the 90 ANTHRAX disease, which had previously been described as independent affec- tions, were all one disease. As late as 1805, Kausch gave a good description of anthrax but denied its contagiousness. Delafond and Gerlach thoroughly investigated ovine anthrax in 1854 and its con- tagiousness was experimentally shown by Gerlach. In 1850, Heu- singer published a very comprehensive treatise on anthrax which deals at length with its history and geographical distribution. Much new information concerning the nature of anthrax was acquired during the fifth decade of the last century. In 1855, Pollander announced the discovery, which he first made in 1849, of minute unbranched rod-shaped bodies in the blood of cattle dead of anthrax. Davaine observed similar bodies in 1850. Then followed a long series of observations and somewhat contro versial discussions on the bacterial origin of the disease, culminating by Robert Koch’s careful description of the morphology of its specific organism including the spore formation in 1876 (1877 Pasteur). Cohn, however, seems to have been the first to have called the organism a Bacillus and to have suspected the existence ‘of spores. Toussaint, in 1880, and Pasteur in 1881, published results of investigations directed toward protective inoculation. Since that time, the literature on the cause, morbid anatomy and prevention of anthrax has become very exten- sive. Geographical distribution. Anthrax is a widely disseminated disease. The continent of Europe has perhaps suffered most from itsravages. It occurs, also, in Northern, Eastern and Central Africa, where in recent years it has become a great plague. In Siberia, it has caused fearful destruction, and in that country it is still known as the “Siberian Plague.” It has frequently appeared in England. Russia, India and Australia are also infected. South America is also reported to suffer much from its ravages. It has occurred in the United States in many localities. There are very few, if any, countries where this disease has not been found. A knowledge of its specific cause, with the methods of properly disposing of dead animals, isolation and disin- fection, as well as the preventive inoculations now in vogue, have made it possible to prevent wide-spread epizodtics. In America it is looked upon as a comparatively rare disease, excepting in certain very restricted infected districts. Etiology. Anthrax is caused by a microdrganism, Bacterium anthracis. This organism can usually be found in the diseased organs ANTHRAX 91 of affected animals. Its spores are very resistant to the normal destructive agencies in nature. Consequently when anthrax is once introduced into a locality it tends to remain there for many years, possibly causing from time to time a few cases or more serious epizodtics, or epidemics. The spores are frequently carried in the wool, hair, hides, hoofs and horns taken from animals sick or dead of anthrax. Through these agencies anthrax has been introduced into distant localities. Bacterium anthracis is a rod-shaped organism varying in length from 1 to + uw, but having a quite uniform breadth of about one micron. In a suitable medium it grows out in long flexible fllaments which com- bine to form thread-like bundles. When examined, the ends of the rod seem to be square cut. In preparations from animal tissues there appear sometimes to be slight concavities in the ends of the segments when two of them are united. In the preparations, capsules are easily recognized. It is believed by Kodama and others that the capsule is a protection to the bacterium against phagocytes but that it does not protect against the bacteriological action of the blood serum. In cultures spores are formed. These are oval, highly refractive bodies held within the cellular envelopes of the filaments, but later they are set free by the dissolution of this membrane. They stain readily with the aniline dyes and also by Gram’s method. The bacterium of anthrax itself is not an especially hardy organism. On the contrary it is easily destroyed by weak disinfectants and it has a low thermal death point. Its spores, however, are among the most hardy of bacterial life to resist chemical and thermal agents. They resist drying for months or years and often boiling for a half-hour or longer does not destroy them. On that account it is very difficult to eliminate the virus from infected pasture lands, especially if they are wet or marshy. Hutyra and Marek state that the spores may pass through the digestive tract without germinating. As the spores may remain on the soil in a dormant condition for many years, it sometimes happens that the disease does not appear until long after the introduction of the virus. Anthrax has been known to break out among cattle grazing on a field in which the carcasses or hides from affected animals were buried many years before. Through some means the spores seem to be able to get to the surface and contaminate the grass. The virus may be introduced with blood or bone fertilizers, hides, hair or wool from infected coun- tries. When the extent of this traffic is realized, it is easy to under- 92 ANTHRAX stand how anthrax has been brought to this country and why it occasionally appears here and there over a large part of the continent. Many outbreaks, as well as isolated cases, illustrating this common method of dissemination are on record. The period of incubation is very short. In inoculated animals it ranges from 1 to 5 days. Channels of infection. Three common modes of infection are recognized for anthrax, namely: through the digestive tract, by the skin and by the lungs. In cattle the infection seems to be largely through the alimentary canal; in horses and sheep by the skin or digestive tract; in men through wounds of the skin and the respira- tory tract. Although these are the usual methods there are many exceptions with each species. Infection through the alimentary canal. This is the more common mode of infection in cattle. The resulting disease has been designated by various names, among which are “intestinal anthrax,” “fodder anthrax,” ‘“‘spontaneous anthrax,” “internal anthrax,’ “anthrax fever,” and anthrax without external manifestations. In these cases the infecting organisms, either the spores or the vegetating bacteria themselves, are taken into the body with food or drinking water. M’Fadyean has shown that infected food-stuffs are often responsible for the infection. It is stated that the infection takes place in most cases in the small intestine, the mucosa of which, it is stated, need not necessarily be injured. It is highly probable that the gastric juice destroys most of the bacteria while the free spores are not injuriously affected by it. In the infected districts, the spores exist at or upon the surface of the soil and possibly on the blades of grass, from which they are easily taken up by grazing animals. In lands thus infected, the specific organism has been introduced at some previous time either by the burying of anthrax animals in these fields, by the use of infected tannery or slaughter house refuse as fertilizers, by flooding from infected streams, or by the bringing of the organism in the droppings of birds or other small animals which have fed upon anthrax carcasses. It is reported that the spores will find their way to the surface even when the dead animals have been buried at a considerable depth. There has been some controversy in the writings of Pasteur, Koch and Bollinger concerning the method by which the spores reach the sur- face. Pasteur supposed that they were brought by earth worms from the buried carcasses. Koch believed this impossible because of the ANTHRAX 93 low temperature of the ground at the depth at which the animals are buried. Bollinger has shown experimentally the possibility of Pasteur’s views. Jarliniski and others have found that the spores of anthrax may be disseminated by slugs, insects and larvae which are found on untanned infected skins. Infection through the skin. In animals, this mode of infection occurs less frequently than in man. Anthrax produced in this way is usually characterized by local manifestations known as “‘carbuncle disease,” or “malignant pustule.” In this mode of infection the bacteria penetrate through wounds in the skin and exposed mucous membranes into the living tissues by means of infected utensils, the use of infected instruments, and insects, especially the house fly (Musca domestica). Dalrymple has called attention to the spread of this disease among animals in the lower Mississippi Valley by means of the horse fly (Tabanidae). In man many cases of the disease occur from injuries or cuts made at the post-mortem of anthrax animals or by the infection of skin wounds while handling infected hides or wool. Maligant pustule is reported to be quite common among the employes of certain tanneries and upholstering establishments where hides and hair imported from infected districts or countries are used. Infection through the respiratory tract. Faser, Buchner, Lemke, and other writers have shown experimentally that the disease can be produced by the inhalation of spores. In man this form of infec- tion is quite common among the wool sorters. In Great Britain, where much foreign wool is handled, it has been reported as causing as many as 500 deaths annually. It is known as “wool-sorters’ disease.” Symptoms. In anthrax, the symptoms vary not only in different species of animals but also in different individuals according to the location of the disease. Again there is often considerable variation when the lesions are apparently the same. The most characteristic features of the disease are the suddenness of the attack, the grave general disturbances, high elevation of temperature, a tendency to ecchymoses of the mucous membranes and local manifestations, such as carbuncles and edema of the skin, digestive disturbances, brain complications and difficult respiration. Anthrax has been classified according to its course as peracute, acute and subacute. It has also been divided according to the site of its manifestations as anthrax with visible localization and anthrax without visible localization. 94 : ANTHRAX Anthrax without visible localization. This form is generally due to ordinary infection presumably by spores. It includes the peracute, acute, and subacute. The peracute or apoplectic anthrax gives rise to symptoms of cere- bral apoplexy. The animal becomes suddenly ill, staggers about for a brief period and falls. There is often a bloody discharge from the mouth, nostrils and anus. Death usually ensues in from a few minutes to an hour. Usually there are convulsions. Sheep and cattle suffer most frequently with this form. They are often found dead. This is especially true in the beginning of an epizootic. In the acute form, the disease runs a somewhat slower course, lasting usually not to exceed twenty-four hours. The temperature rises rapidly to from 105 to 108° F., dropping suddenly just before death. With this there are signs of congestion either of the brain or of the lungs. If the brain is affected the animal becomes restless, excited, stamps the ground, rears in the air, bellows, runs to and fro and finally goes into convulsions followed by stupor and death. If the lungs are congested there is difficulty in breathing, more or less wheezing, panting, groaning, palpitation of the heart, small and fre- quent pulse, cyanosis of the mucosa of the head, bloody discharges, hematuria, staggering and finally convulsions and death from suffoca- tion. Occasionally there is a partial remission of the symptoms, followed by relapse. It has been observed that occasionally there are premonitory symptoms preceding the acute attack consisting of slight digestive disturbances and diminished vivacity. Burnett found the anthrax bacteria in large numbers in the blood during this stage. He likewise found them to be present in the blood of the more chronic cases during the febrile period. The subacute form is known as anthrax fever or intermittent an- thrax. The symptoms are the same as in the other forms, except that they are more sharply defined and the course is longer. The disease lasts from one to seven or eight days. The high temperature, the congestion of the lungs or brain complicated with intestinal distur- bances, especially colic, are usually well marked. In epizodtics where the peracute or acute form ushers in the disease, the later cases usually are of the subacute variety. Anthrax with visible localization. These forms usually result from infection of the skin and mucous membranes. This form is common in horses and sometimes it occurs in cattle. It is reported to occur in other species. The lesions are circumscribed, cutaneous swellings d ANTHRAX 95 which are at first hard, hot and painful. Later they become cold and painless, with a tendency to become gangrenous. The edematous tissue becomes doughy, cold to the touch and painless. Frequently fluctuating swellings of the skin occur. The duration of this form of the disease varies from four to fifteen days. Ordinarily it is not so fatal as internal anthrax. When the infection is on the mucous membrane of the mouth or pharynx the animal suffers from fever, dyspnea, difficulty in swallow- ing and cyanosis, together with the immediate local effects. Death occurs much sooner than when the disease is located in the skin. It is stated that dogs and swine suffer from this form more than from the acute types. In horses, anthrax usually runs an acute or subacute course. The first symptom is rise of temperature with a rapid, feeble pulse. There may be chills and muscular spasms. The mucosa of the head becomes cyanotic and lacrymation is often present. ‘The animal has a dull, stupid look, appears to be stunned and walks with a staggering gait. In some cases there are symptoms of cerebral congestion,, such as restlessness or convulsions. Colic is a very characteristic symptom in the horse, otherwise the symptoms are the same as in cattle. Infec- tion of the skin usually occurs on the hypogastrium, lower part of the breast, inner surface of the fore and hind quarters. Swelling of the hind quarters often causes lameness. In sheep and goats the disease is usually of the acute or apoplectic form. The animals appear as if suddenly stricken with apoplexy. If death does not occur within a very short time, symptoms already described for this form of the disease may be recognized. Subacute -anthrax is said to be very rare in sheep. In swine, anthrax is ordinarily characterized by local lesions on the mucous membrane of the larynx and pharynx. The animals have a rise of temperature and the intermaxillary space is generally swollen. The swelling may spread along the trachea, giving rise to difficulty in swallowing, hoarseness, cyanosis of the mucosa of the mouth, dyspnea and rapid breathing. The animal shows signs of paralysis. Death occurs from suffocation. Frequently the tongue becomes the seat of the disease. In dogs and cats, the disease usually runs a very rapid course. The fact that they are usually infected by eating the meat of animals dead of anthrax causes them to suffer largely from the intestinal form. It 96 ANTHRAX has been stated that probably much of the so-called anthrax in dogs may be ptomaine poisoning. It is reported that in birds anthrax usually runs a very rapid and usually fatal course. Toward the end they stagger, tremble, or go into convulsions and die with bloody discharges from the mouth, nostrils and anus. From the first the birds are depressed, weak, and their feathers ruffled. There is evidence of dyspnea. Carbuncles are said to appear on the comb, wattles, conjunctiva, tongue and extremi- ties. Jt has been stated that the milk from cows suffering with anthrax contains Bact. anthracis. Moore found in the examinations made in one epizoétic that they were present in considerable numbers in the milk just before or immediately after death, but they were not found in the milk of animals in the earlier stages of the disease. The duration varies from a few hours to a week or even longer. The prognosis is unfavorable. Jn some herds the mortality is 100 per cent. while in others a number of animals may recover.* The average mortality is placed at about 70 per cent. in animals. In the human species many persons recover from its local form (malignant pustule). Morbid anatomy. The nature and extent of the tissue changes depend upon the course of the disease. When experimentally pro- duced it is ordinarily a septicemia. This form often occurs in domes- ticated animals when they contract the disease naturally. The more common anatomical changes, except in the most acute cases and in the strictly localized lesions or carbuncles, are: Hemorrhages varying in amount from petechie to blood extravasa- tions, with more or less serous, gelatinous and hemorrhagic infiltration of the submucous, subserous and subcutaneous tissue. The capillaries are distended and frequently there are hemorrhages due to changes in the walls of the capillaries. The anthrax bacteria are often present in large numbers in the smaller blood-vessels. It is believed by many that the capsules absorb much of the body liquid, *M’Fadyean has reported this disease in 39 consecutive outbreaks in which a total of 54 animals died. In New York the disease existed in 1904 in 15 herds in one locality. There were more than 30 deaths. In one herd of 21 animals, 20 had the disease, 16 died and 4 recovered. In another dairy 4 out of 7 died, but in the others one or two animals in each were affected. In 1903 anthrax occurred on 84 different farms in the same county. There were 170 fatal cases of which 33 were in horses, 123 in cattle, 11 in sheep, and 3 in hogs (Burnett). These facts are interesting in showing that the disease does not always cause heavy losses in the infected herds. ANTHRAX 3 97 causing them to swell and preventing the fluid from reentering the circulation. This gives rise to the edematous swellings. The sub- cutis may be sprinkled with ecchymoses. Frequently there are gelatinous effusions of a rather firm consistency and of varying size. The color also differs, ranging between a deep yellow and a yellowish brown. Often these edematous areas are sprinkled with hemorrhagic foci. A simple serous edema may occur. The lymphatic glands may be hemorrhagic, edematous or both. An edema of the connective tissues of the neck or about the trachea is often very marked. The muscles vary in color but usually they are darker than normal, and, like the skin, they often become sprinkled with ecchymoses. The heart muscle suffers from parenchymatous changes. In the larger cavities of the body, a sanguinolent fluid is found in moderate quantities. Blood extravasations of different sizes are seen under the serous membranes, particularly on the mesentery and mediastinum. The subserous connective tissue, especially on the mesentery, anterior mediastinum and in the neighborhood of the kidneys, is often infiltrated with a gelatinous substance. On this account the neighboring lymph glands are considerably swollen, filled with serum and sprinkled with hemorrhages. The internal organs contain a large quantity of blood. All the larger veins and the heart are filled, while the surrounding tissues show sanious imbibition. The spleen is usually considerably enlarged (two to five times its normal size), either uniformly or by prominent tumor-like protuberan- ces. The pulp is soft, more or less fluid, and stained a dark-red color. The capsule is always very tense. It is frequently sprinkled with ecchymoses. Occasionally this organ is slightly affected. The liver and kidneys are highly congested and somewhat enlarged. The parenchyma contains areas of blood infiltration and the cells themselves manifest various kinds of degeneration. The portal lymph glands often appear to be enlarged, and the retroperitoneal tissue may be infiltrated with a serous, gelatinous fluid. The sub- peritoneal tissue of the intestines and of the abdominal walls may be similarly affected. The nature of the lesions of the intestinal canal varies according as the disease is intestinal anthrax, or anthrax caused by inoculation. In case of inoculation anthrax, the intestine is frequently normal. In other cases there may be submucous and subserous hemorrhages, or swelling of the mesentric glands. The principal changes in intestinal 98 ‘ ANTHRAX anthrax are always found in the small intestine, chiefly in the duo- denum. In the milder cases of intestinal anthrax the mucous mem- brane is affected by circumscribed or diffuse swellings. The bacteria are often found in very large numbers on the surface of the mucous membrane. Necroses and ulcers appear in those parts where the bacteria are most thickly congregated. In very severe cases, the abomasum or the true stomach may be affected with a gelatinous and sanious infiltration of the mucous membrane. The mucosa of the abomasum, and especially of the duodenum, is, in consequence of excessive hyperemia, dark-red or almost black, and is covered with erosions and ulcers or necroses, which may extend down to the sub- mucosa. The contents of the intestine are bloody, and the submucosa is infiltrated with a serous, gelatinous, or hemorrhagic exudate, so that the mucous membrane often projects in the form of large tumors into the lumen of the intestine. On the site of Peyer’s patches and the solitary follicles we may find flat or prominent nodules, the surface of which are covered with diphtheritic crusts. The lungs are greatly congested, edematous and show areas of ecchymoses. The entire respiratory mucous membrane is consider- ably reddened and ecchymotic. The mucous membrane of the pharynx and opening of the larynx is often so edematous that stenosis of the larynx takes place. The contents of the trachea and the bronchi consist mostly of bloody froth or mucus. The brain is often studded with ecchymoses. The surface of its membranes often exhibits hemorrhages with an accumulation of sanious serum in the ventricles. Extravasations of blood sometimes occur in the anterior chamber of the eye and under the retina. All the other organs may contain hemorrhages, and the urine frequently contains blood. The blood is usually dark. It has a tarry or varnish-like lustre, and shows little tendency to coagulate. It does not assume its normal red color when exposed to the air. Burnett studied the blood of a few cases of anthrax in 1904. The appended tables are taken from the results of his examinations. ANTHRAX 99 RESULT OF THE EXAMINATION OF THE BLOOD OF FIVE CASES OF ANTHRAX IN CATTLE foe Leen See) SE ew ee le cbserved | tion | 3° Per | pere.mm. | Per No. 8 | July 8 [tors "Anth, bact. in blood “8 July 9 | 106.6 | 60 | 4,072,000 "20,000 Died July 9 m U “ 4*) July 10 ‘106.2 | “4 “July rl 56 | 5,471,000 | asi a [oe | 108.0 38 | 3,400,000 | 3,444 “4 | «19 56 2,086,000 | 9,876| Recovered “ st July 7 | Anth. bact. in blood “ 6 July 91040] 50 8,876,600 | 8,222 “« 6 Som 60 8,954,000 5,210 6 * 13 | 101.8 | BEG “14 | 101.2 47 3,484,000 | 5,666, Apparently recovered. “ 6 “19 54 | 1,980,000 a7 Died of anthrax Nov. 4 “ 6 “94 63 | 3,132,000 “11,888 “ 3 jJune 29 | 68 July 14 | 101.2 | 57 | 4,168,000 5,222 Recovered | ey | ety 15 | ae July 16 | 103.8 | 53 | 2,324,000 8,111 { ae | “ 17] 101.0 58 | 2,632,000 | 5,333 «yl * 1g | 102.0 ' 8,163 aT «19 5,940,000 | 11,000 ae | “Od 61 10,767 Recovered | | *Temperature July 8, 102.1° +Temperature July 8, 107.4° Temperature July 10, 100.0° Temperature July 9, 103.0° 100 ANTHRAX THE DIFFERENTIAL COUNT OF THE LEUCOCYTES IN FIVE CASES OF ANTHRAX IN CATTLE Cow | Pate $8 | Lymphocytes Large Mono} pojynuclear |‘Eosinophiles| Mast 88 | No. | 0% | No. | 96 | No. | 96 | No. | 0% | No. | 06 Soper No. 8 | July 9} 20000 | 7080 | 35.4 2200 | 11.0| 7120 | 35.6 | 3520 | 17.6 | 80 | 0.4 “ 4) © 410 QA 52 64.8 28 0.2 «4/ “ a1 4814 | 1670 | 34.7! 341 | 7.1] 2200 | 45.7] 548 | 11.4 | 48 } 1.0 ode ia 18 3444 | 1482 | 41.6] 261 | 7.6] 1667 | 48.4| 75] 22] 7102 4 | 19, 9876 | 4957 | 43.1] 273 | 2.7| 4606 | 47.5| 636] 6.4| 11 |0.2 reg | 9 8222 | 3930 47.8| 296 | 3.6| 3436 | 41.8] 518] 6.3 lat los 6 @| ss uy 5210 | 2287 | 43.9| 338 | 6.5] 2115 | 40.6| 432] 8.3 | 36 |0.7 oe) ee | 40.1 4.0 47.2 78 0.7 ee 4 5666 | 2898 50.0] 119 | 2.1] 2221 | 39.2} 430] 7.6 | 62/14 © 4 | “49 777 | 9747 42.7| 263 | 3.0| 2800 | 31.9 | 1930 | 22.0 | 35 |0.4 “6) “ 24 11888 | 6033 | 50.7| 698 | 5.8| 3120 26.2| 1946 16.3 | 89 |0.7 “3) « ql 5292 | 3352 | 64.2| 261] 5.0| 1451 278 146| 2.8 | 10 |0.2 Bradish No.1| “ 16} 8111 | 3598 | 43.5] 154] 1.9] 3033 |37.4 | 1330| 16.4 | 65 |0.8 4 | “ 17] 5333 | age 53.1 | 256 | 48] 1568 | 29.4] 640| 12.0 | 36 |0.7 “1/1 " 18) 163 | 6375 7s.1| 138 | 1.7{ 914] 11.2] 734] 9.0 a | “ 19) 11000 | 6611 60.1 | a53 | 2.3| 2585 | 23.5| 1496| 13.6 | 55 [0.5 “ 1) gs! 10767 | 5911 st. 484 | 4.5] 2186 | 20.3] 2099 | 19.5 | 86 |0.8 ANTHRAX 101 Burnett found that the number of red corpuscles and the percentage of hemoglobin are reduced. In the chronic cases they tend to return to the normal condition. There was an increase in the number of lymphocytes and a decrease in the number of polynuclear leucocytes. In some cases there was a marked increase in the number of eosino- philes. No change from the normal was noted in the large mononu- clear leucocytes or in the mast cells. The bodies of animals which have died from anthrax are often weil nourished. Rigor mortis is absent and they decompose quickly. Very frequently blood flows from the natural openings of the body, and the rectum is sometimes prolapsed. All the foregoing lesions may be absent in very acute apoplectic cases. The specific organism is, however, always present in the cadaver. It is important to note that occasionally the usual changes indicated by the symptoms and the duration of the disease are not found on post-mortem examination. In one epizodtic, the writer saw an animal dead from subacute anthrax in which the blood and tissues were teeming with anthrax bacteria, yet the organs examined microscopically appeared to be normal. Other animals in the same outbreak exhibited the more usual anatomical changes. Diagnosis.* There are a number of methods for diagnosing an- thrax. The symptoms and lesions are of value but often they are not sufficiently characteristic to enable one to make a positive determina- tion. This must rest on the bacteriological examination and the specific reactions. Bacteriological. It was believed for many years that the bac- teriological examination for the diagnosis of anthrax was very simple and sure. That opinion is entertained by many veterinarians still. The facts are that many cases can be readily identified by this method if the tissues are fresh, but, on the other hand, there are those where it is impossible. The difficulties lie in several directions. The most important is the rapidity with which anthrax bacteria die in tissues where sporulation does not occur. For this reason it is not always possible to find the organisms in specimens sent to a laboratory. The *Veterinarians should recognize that anthrax is often very difficult to diagnose in the laboratory. The lesions are frequently localized and in such cases the specific organism is not always present in the general circulation or in other organs. In such cases the man who makes the post mortem examination must locate the lesions and select parts of the affected tissues for bacteriological examination. While the typical cases are readily detected, there are those where the limitations of the laboratory methods and the neglect cf the clinician in making the post mortem allow the disease to go undetected. 102 ANTHRAX careful work of Fischoeder has clearly pointed out the difficulties in this direction. Secondly, errors may occur because of the presence of what are called pseudo-anthrax bacteria from which a differentiation is not alwayseasy. Fitch pointed out that these organisms could not be differentiated from Bact. anthracis from agar or gelatin cultures. Pokschischewsky has studied their biology and shown their very close resemblance, in certain particulars, to Bact. anthracis. He divides these organisms according to their growth on agar, gelatin and potato. into two types, namely, pseudo-anthrax and anthraxoid. Microscop- ically the presence of an organism resembling that of anthrax, often found in tissues some hours old, may be mistaken for that of anthrax. The diagnosis bacteriologically requires the isolation of the specific bacterium and its identification by cultural methods or animal inocu- lation. Differential stain. M’Fadyean has described a peculiar staining reaction, first pointed out by Heins, which he considers of value for the microscopic diagnosis of this disease. The reaction is in evidence when films of blood, exudates, or tissue juice containing the bacteria are stained with a simple aqueous solution of methylene blue. The method as applied to blood is as follows: Place a drop of the blood on a clean slide. The size of the drop should be about 2 mm. in diameter. It is spread quickly with a platinum needle untilit covers an area about 12 mm. in diameter. Protect from dust and allow the slide to remain until all evidence of moisture has disappeared. When dry, heat the preparation by lowering it film upwards into the flame of a Bunsen burner or an alcohol lamp for a second. Repeat this three times or until the glass is too hot to be borne by the skin in the palm. of the hand. Allow the slide to cool and then cover the film with 1 per cent. aqueous solution of methylene blue. After a few seconds pour off the free stain and wash the slide thoroughly in tap water. Dry the slide by pressing it gently between two layers of bibulous paper, and then more thoroughly by holding it in the current of hot air above the Bunsen flame. Finally, mount in Canada balsam. The microscopic examinations (x 800 to 1000) will show an occasional leucocyte and the anthrax bacteria. There will appear no other visible formed elements. The nuclei of the corpuscles generally exhibit a greenish-blue tint, the anthrax rods are stained blue. The intensity of the stain depends upon the length of time after death before the films were made. Usually the segment character of all but the shortest rods will be apparent. If they are deeply stained this is not very distinct. The peculiarity in the reaction lies in the color of the amorphous material which is present between and around the bacteria. This material presents itself under the form of coarse or fine granules of a violet or reddish-purple color, which is in sharp contrast to the tint of the bacteria or cell nuclei, especially with brilliant lamp or gas light. These violet granules differ a good deal in form and size; sometimes they are very minute, and at others coarsely granular. When the bacteria are arranged in clumps the violet material is often in ANTHRAX 103 greatest amount about them. Free-lying anthrax rods will be surrounded by a thick envelope of the same substance. M’Fadyean states that he has never found this reac- tion in animals dead from other diseases. The peculiar coloring, he states, in some cases may be observed without the aid of the microscope. Our experience with it has not been so satisfactory. Thermoprecipitation. This method was formulated for practical work by Schutz and Pfeiler. It was especially advocated by Ascoli for the diagnosis of anthrax. It has been tested by a number of laboratory workers and as a rule highly recommended. The method is based on the fact that anthrax bacteria, or their decomposition products, present in the bodies of animals dead of anthrax, contain a precipitinogen which, when brought into contact with anthrax immune serum, produces a precipitate at the point of contact. Pickens describes the method in detail and concludes that it is a reliable means for the diagnosis of anthrax. It was found, however, by Pokschischewsky that this reaction took place in cases of infection with certain of the pseudo-anthrax organisms. Method—tThe principle of this test is based on the fact pointed out by Kraus in 1897, that an immune serum when brought into contact with its corresponding antigen will produce a precipitation at the point of union of the two fluids. The first fluid, the immune serum, is difficult to produce. The technic was first worked out by Ascoli. The selection of the animal for the production of this serum is an important problem to decide. According to Ascoli, Schutz and Pfeiler and others, the ass is the most desirable animal. However, good sera have been produced from the horse, mule, cow and rabbit. Varying degrees of success have been obtained with the dog, sheep, goat and guinea pig. The selection of the animal is important as only certain individuals produce a proper serum. It is stated by Schutz and Pfeiler that out of one lot of thirty animals only three were found that produced a satisfactory serum. According to Ascoli and others, the serum of certain normal animals, especially the horse, is apt to produce a precipita- tion when brought into contact with salt solution, carbol salt solution, distilled water or bouillon. The serum of the animal to be immunized should be tested with reference to this quality. The test requires an immune serum and the antigen. The animals are immunized by repeated injections of attenuated, slightly virulent and virulent cultures of Bact. anthracis beginning with the attenuated culture. Several injections are necessary. They may be made subcutaneously, into the abdominal cavity or intravenously. The injections are made about a week apart. It requires several weeks to obtain a satis- factory serum. The blood should not be drawn from the immunized animal for at least ten days after the last injection. To obtain the serum the blood is drawn in the usual way and allowed to clot and the clear serum drawn off. The serum must be perfectly clear and not colored with hemoglobin. 104 ANTHRAX PREPARATION OF THE ANTIGEN There are three different methods by which an antigen may be prepared from the suspected material, namely: The boiled extract, the shake extract and the slow process. He also describes a method for the preparation of an antigen made from a culture of the anthrax organism. This antigen should be used as a control upon the material to be tested by this reaction. The Boiled Extract. This extract consists in taking a small piece of the material to be examined and placing it in a test tube, and adding to it four or five volumes of normal salt solution. The test tube is then placed in boiling water from five to fifteen minutes. This yields a cloudy brownish fluid which may be cleared by filtering through filter paper. Two or more filtrations may be necessary. According to Schutz and Pfeiler, this antigen may be improved by the use of carbol salt solution instead of normal salt solution. They state that the carbolic acid will also preserve the extract so that it may be kept for later use. The Shake Extract. This extract consists in placing a piece of tissue about the size of a hazelnut in a test tube to which is added about 10 ¢.c. of normal salt solution. It is then thoroughly shaken. This yields a dirty, reddish fluid, which is centrifuged for clearing purposes. It is then filtered through filter paper or asbestos. This process may have to be repeated several times to obtain a clear antigen. The Slow Process. This consists in taking a piece of suspected material about the size of a hazelnut and triturating in a mortar with about ten grams of dry, white sand. To this is added enough chloroform to completely cover it. The chloroform is allowed to remain for several hours, after which it is poured off. The residue should then be thoroughly stirred with a glass rod and enough normal salt solution added to cover it. This should be stirred again and the fluid filtered into a test-tube through ordinary filter paper. The filtrate should be colorless or slightly yellow. If it is not clear, it should be refiltered until it is. The chloroform does not have any effect upon the mixture except to precipitate the hemoglobin. The precipitating substance is not soluble in chloroform, and hence the final extract is not weakened by this process. In cases where the organs contain only a few bacteria, a longer extraction by the normal salt solution may be necessary. The Culture Extract. For this extract, a 24-hour agar culture of the anthrax organism is used. Five to 10 c.c. of normal salt solution is poured over the surface of the culture and allowed to act for two hours at room temperature. At the expiration of that time, the fluid is removed and filtered through ordinary filter paper. The filtrate must be clear, or slightly yellowish in color. Schutz and Pfeiler again prefer the carbol salt solution to the normal salt, as they think it produces a stronger antigen. Of the three different methods for the preparation of the antigen, Ascoli favors the boiled extract. On the other hand Schutz, Pfeiler and Pickens believe the slow process to be the best. They contend that an antigen prepared by this method contains more precipitinogen than by either of the other processes. -For the preparation of the antigen, any of the tissues of the body may be used. The spleen, blood, serous or hemorrhagic exudates are preferable and in the order named. Of the remaining organs, the liver, lungs, kidney, muscle and skin should be men- tioned. In case the skin is used, plenty of subcutaneous tissue should be included with it. According to Negroni, the presence of anthrax may be detected in imported skins by this method. ANTHRAX 105 The quality of the immune serum is roughly estimated as follows: The serum that will produce a reaction immediately when brought into contact with an anthrax antigen is considered univalent. It has been learned that good sera will show the presence of an anthrax antigen in a normal unit and, also, in fractions of the same. A serum that reacts to an antigen diluted 100 times is a hundred fold serum. This is called a standard serum. In testing the material for the presence of anthrax by the Ascoli reaction, it is not known whether the precipitinogen is present or not, hence its titration is impossible. But in case of the titration of the immune serum where the antigen is prepared from known anthrax material, some kind of a standard is necessary. For this standard, the culture extract is used. TECHNIC OF THE TEST According to Ascoli, the serum should first be placed in a test tube after which the extract, the lighter of the two fluids, should be placed beneath it by means of a capillary pipette. According to Schutz and Pfeiler, just as good results are obtained by placing the heavier fluid, the immune serum, in the tube first and then placing the antigen on top of it. They use thesame technic used by Pfeiler in his diagnosis of glanders by the precipitation reaction. With an ordinary pipette, they place the serum drop by drop on the edge of a small test tube, about 6 millimeters in diameter and 12 centimeters long. The serum is allowed to run slowly down the inside of the tube to the bottom. Enough serum should be added to bring the top of the fluid up about one-half a centi- meter in height. After the serum has reached the bottom and remains stationary, the antigen should be placed on the edge of the tube in the same manner as the serum, and allowed to run slowly down the tube until it reaches the serum. It is advantageous to have the antigen take the same course down the tube as the serum. When the extract reaches the serum it should form a sharply defined layer above it. If the extract mixes with the serum, at the point of contact, the test will be ruined. However, if the extract does not mix with the serum, and the two fluids form, at the point of contact, a sharply defined division line, the technic has been properly executed. In case the antigen was prepared from anthrax material, a cloudy, grayish white ring will form immediately at the point of contact of the two fluids. This ring gradually increases in density for some time. It may be seen best in an oblique light. The test tubes used in this reac- tion do not necessarily have to be sterile, but it is imperative that they should be scrupulously clean, for if they are not clear the ring may not be seen. In case the antigen is prepared from material taken from an animal that did not have anthrax, this ring will not occur, or if it does occur will not take place under fifteen minutes. In case the reaction is positive, the ring remains visible for two hours, and longer, after which it disappears entirely. A test, however, may be read on the suc- ceeding day by the presence of sediment in the bottom of the tubes containing the positive reaction. The tubes containing negative reactions will show no precipitation. Control. Schutz and Pfeiler state that with reliable sera the control tests may seem superfluous, but still they do not wish to discard them. They make control tests only with suspicious material, and not with material known to be negative or positive. They make the statement that the extract, in every case, should be tested with normal serum from the same species of animal from which the antigen originates. They also use a positive and negative control for their antigen in routine work. In all, this makes three controls which are, the normal serum, the anthrax extract and the normal organ extract. 106 ANTHRAX Anthrax is to be differentiated from certain specific diseases such as symptomatic anthrax (black leg), malignant edema, and septicemia hemorrhagica. Rabies is not infrequently mistaken for anthrax. These diseases can as a rule be readily diagnosed by the methods applicable to each. In addition to the specific infections, anthrax has been confused with certain dietary troubles and poisoning. Protective inoculation. Toussaint was the first to make use of protective inoculations in anthrax. He heated defibrinated blood to a temperature of 55° C. for 10 minutes. Better results were obtained by heating the blood to 60° C. for 3 or 4 times before using it. Pasteur, however, was the first to prove that immunity could be obtained by the use of cultures of attenuated bacteria. Several methods of attenuating the specific organisms were proposed by Pasteur, Tous- saint, Chaveau, Chamberland, Arloing and others. Pasteur’s method consists in inoculating the animal with a small quantity of culture which has been grown at a high temperature— 42 to 43° C.—_for several days. This deprives the bacteria of their virulence. To strengthen the resistance, the animals are again inoculated 12 days later with a stronger virus.* After the two inocu- lations, they are said to be protected against the most virulent an- thrax; but the immunity is of short duration. Chamberland reported in 1894 that a total of 1,988,677 animals were treated by this method in France, and that the loss from anthrax had diminished from 10 per cent. in sheep and 5 per cent. in cattle to less than 1 per cent. Cope, in his report to the English Board of Agriculture, regards the con- clusions of Chamberland 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. The excellent work which has been done by Neal and Chester, at the Delaware College Experiment Station, has shown the possible efficiency of this method. Of the 331 cows which they vac- cinated against anthrax, two died of the disease, giving a death rate of less than 1 per cent. and this in a territory so saturated with the virus that it was practically impossible to keep cattle at all before its *The first vaccine is a culture of anthrax bacteria that has been cultivated so that it will kill mice but has no ill effect on rabbits and sheep. The second vaccine, given 12 days later, is more virulent. It will kill mice and guinea-pigs and occasionally rabbits. Immunity is established 12 days after the second inoculation. The French recommend the following plan of injection. The first vaccine is injected into the internal surface of the right thigh and the second into the internal surface of the left thigh. The vaccine should be used as soon as it is procured. A contaminated vaccine should not be injected. ANTHRAX 107 use. The objection to this method is, that it requires the use of the living bacteria, which later may become virulent and consequently cause a subsequent outbreak. The scattering of pathogenic organ- isms, even in an attenuated condition, should be avoided if possible. It must be admitted, however, that Pasteur’s method has done much good and helped to rob anthrax of much of its former terror, especially for the farmers of Europe. In America the spread of anthrax has been checked in many districts by its use. Dalrymple has pointed out its success in the lower Mississippi Valley. Chester and Neal used it successfully in Delaware. They pointed out that a vaccine which succeeded at one time proved fatal at a subsequent time. Notwithstanding, it is highly probable that the spreading of a knowledge of the specific cause of this disease with instructions for the proper disposition of dead animals has also exerted much influence for good in checking its ravages. In Germany and England the stamping-out system is considered superior to vaccination. According to Crookshank, in England it is regarded as the only reliable means of suppressing the disease. To this end rigid laws have been enacted. In this country as rigid measures as possible for its eradication should accompany the use of methods for establishing a tolerance for its existence. The simultaneous method. This method consists in the injection of anthrax serum* together with a small quantity of virulent anthrax bacteria. It has proven to be very satisfactory. It has the advan- tage of being administered at one time. This method of protection against anthrax seems to have been first proposed by Sobernheim in 1899. He reports excellent results from its use in immunizing cattle and sheep against anthrax in South America. Sclavo has produced serums which seem to have a therapeutic as well as prophylactic value. It has been used in treating human anthrax since 1897. Prevention. In all cases the well animals should be removed from the barns or yards containing the sick ones and from pasture lands on which the sick became infected. The temperature of the apparently healthy animals should be taken morning and evening from one to two weeks after they are removed and all of those showing an eleva- tion of temperature should be isolated. By careful isolation and safe *Horses are immunized in from 10 to 12 days. They are injected with 5 cc. of the serum and are from 0.25 to 0.30 cc. of a culture of anthrax bacteria in different parts of the body. According to Sobernheim the successful use of the serum depends upon using the virus in combinations with it. t 108 ANTHRAX disposition of the dead animals the spread of the disease can be checked. Animals do not, as a rule, spread the vitus when the first symptom (rise of temperature) can be detected. All infected stables and yards should be thoroughly disinfected. The disposition of dead animals in an outbreak of anthrax is a matter of much importance. In all cases they should be burned if possible, if not, they should be deeply buried and covered with quick lime before the dirt is replaced. The ground over the place where they are buried should be fenced in to prevent other animals from grazing over it, and the surface should be burned annually for some years to destroy spores should they be brought to the surface. Control. Owing to the long resistance of the anthrax spores, it is impossible to render an infected field safe for pasture land. After a number of years such fields may become harmless. The safe disposal of all carcasses by burying deep or cremation; the thorough disinfec- tion of all stables and paddocks; and the burning of all litter that might have become infected are precautions that should be taken. In localities where there are frequent outbreaks of the disease regular vaccination of all cattle, horses, sheep and swine is advocated. It is all that can be done beyond preventing exposure to known in- fected localities. Anthrax should be reported but a quarantine against healthy animals does not seem to be necessary as the disease is not ordinarily communicated directly from one animal to another. There is a possibility of removing animals while in the period of incubation, thereby infecting other places. Little is known about the “carriers’’ in this disease. REFERENCES 1. Ascoli. Die Prazipitindiagnose bei Milzbrand. Centralbl, f. Bakt. u. Parasitenk., Bd. LVIII (1811), S. 63. 2. Burnett. The control of an outbreak of anthrax. Am. Vet. Review, Vol. XXXII (1908), p. 136. 3. Custer. Anthrax, bacteriological work. Report Del. Agr. Expt. Station, 1895, p. 64. 4. Cuerster. Protective inoculation against anthrax. Proceedings of the Society for the Promotion of Agricultural Science, 1896, p. 52. 5. Datrympte. Anthrax and protective inoculation in Louisiana. Proceedings of the Am. Vet. Med. Assn., 1901, p. 147. 6. Davartne. Recherches sur les infusoires du sang dans la maladies connue sous le nom de sang de rate. Compt. Rend. de l’ Acad. des Sc., 1863, 1864, 1865. 7. ErcuHorn. Experiments in vaccination against anthrax. Bulletin No. 340, U.S. Dept. of Agric., 1915. 8. Fiscnorper. Berliner Tierdrztliche Wochenschrift, Bd. X XIX (1913), Nos. 36, 37, 38 GLANDERS 109 9. Fircu. Organisms morphologically resembling anthrax bacteria. Report N.Y. State Vet. College at Cornell University, 1909-10, p. 200. 10. Kocn. Die Aetiologie der Milzbrand-Krankheit begriindet auf die Entwicke- peperndite des Bacillus Anthracis. Cohn’s Beitr. zur Biol. der Pflanzen, Bd. II 76), S. 277. 11. Kopama. Ursache der nattrlichen Immunitat gegen Milzbrandbacillen. Centralbl. f. Bakt. u. Parasitenk., Bd. LX VIII (1913), S. 373. 12. M’Fapyean. Anthrax. Jour. Compar. Path. and Therap., Vol. XI (1898), p. 51. 13. M’Fapyean. A peculiar staining reaction of the blood of animals dead of anthrax. Jour. of Compar. Path. and Therap., Vol. XVI (1903), p. 35. 14. M’Fapyrean. Extraneous sources of infection in outbreaks of anthrax. Jour. Compar. Path. and Therap., Vol. XVI, p. 346. 15. Moore. Report of an outbreak of anthrax. Annual Report, Commissioner of Agriculture of the State of New York. 1897, p. 550. 16. Pasteur, CHAMBERLAND ET Roux. De l’attenuation des virus et de leur retour & la virulence. Comp. Rend. d. Acad. des Sc., Vol. XCII (1881), p. 427. 17. Pasteur. La vaccin du charbon. Ibid. p. 666. 18. Pickens. The determination of anthrax by means of the thermo-precipita- tion reaction. Report New York State Veterinary College at Cornell University, 1914, p. 220. 19. Poxscuiscuewsky. Uber die Biologie der Pseudomilzbrandbacillen Beitrage zur Differentialdiagnose der Milzbrand und Pseudomilzbrand bazillen. Arbeitena.d. Kaiserlichen Gesundheitsamte, Bd. XLVII, S$ 541. 20. Scnutz anp Premner. Der Nachweis des Milzbrandes mittels der Prazipi- tationsmethode. Archiv. f. wissen. u. prakt. Tierheilkunde, Bd. XXXVIII (1912). 21. Russevt. Outbreak of anthrax fever traceable to tannery refuse. The 17th Annual report of the Wis. Agric. Exp. Station, 1889. 22. Sopernuermm. Ueber das Milzbrandserum und seine praktische Anwendung. Deut. med. Wochenschr., 1904. No. 26 u. 27. (First publication. Zeit. feir Hygiene, 1899, Bd. XXXI). GLANDERS Synonyms. Malleus; farcy; morve: Rotzkrankheit. Characterization. Glanders is one of the most important diseases of horses, asses and mules. It is communicable to man. It runs an acute or chronic course, attacking the lymphatic system more espec- ially in the upper air passages, lungs or skin. The disease is charac- terized by a strong tendency to the formation of small neoplasms or nodules which are likely to degenerate into ulcers from which exudes a peculiar sticky discharge. In the very acute cases a considerable rise of temperature and general debility may accompany the forma- tion of the lesions. Glanders of the skin is known as farcy. By direct inoculation several species of animals may be infected. Thus the disease has been reported in goats, rabbits, sheep, guinea pigs, field mice, and several of the wild animals, especially those of the cat tribe. Swine and pigeons are very slightly susceptible. Cattle, white mice, rats and domestic fowls seem to be immune. 110 GLANDERS History. Glanders is reported to have been known long before the Christian Era. The name malleus was given to it by Aristotle. The theory of the contagiousness of glanders was much doubted at the beginning of the last century. The view taken by the veterinarians at the Alfort Veterinary College was that glanders might arise spon- taneously from an attack of strangles. This view was far more widely accepted than the theory of its contagiousness, which was stoutly sup- ported by the authorities at the Veterinary College of Lyons. It was not until Rayer (1837) had demonstrated the transmissibility of glanders to man, and Chauveau (1868) had shown that the virus was contained chiefly in the firm component parts of the infective material, that the fact of the infectious nature of the disease was accepted. The theory of the spontaneous ‘origin of glanders was widely accepted in Germany. It was believed that glanders could be pro- duced by the injection of pus, and that strangles could develop into glanders. Glanders was looked upon as a tubercular disease, scrofula, pyemia, diphtheritis, general dyscrasia and cachexia respectively. Virchow was the first to declare that the nodules of glanders were independent, anatomical formations, which he placed under the head- ing of granulation tumors. Gerlach was the strong advocate tor the exclusively infectious origin of the disease. Leisering appears to have been the first to give an accurate description of the lesions. The first biological researches into its nature were made in 1868 by Zurn and Hallier, who found a fungus which they believed to be its cause. In 1882, Loeffler and Schiitz succeeded in finding the bac- terium of glanders, in cultivating it, and in transmitting the disease to other animals by inoculating them with pure cultures of the organism. Their researches furnished the positive proof that glanders is a specific, infectious disease, produced exclusively by Bacterium mallet. Geographical distribution. Glanders exists in the greater part of the civilized world. It is more common in the temperate zones, where traffic in horses is active. In the United States it was largely con- fined to the Northern States before 1861, but it spread over the South in connection with the civil war. It is said to have entered Mexico with the American cavalry in 1847. Similarly, Portugal is said to have been exempt until the invasion by Napoleon in 1797. Central Hindoostan was said to be free from it until the war with Afghanistan in 1878. In all these cases, the movements of cavalry, artillery and of commissary trains were responsible for the introduction of the GLANDERS 111 disease into new territory. In our own case the sale of horses and mules at the close of the civil war produced a very general diffusion of this disease, from which the country is still suffering. Insular places, especially if far from the main land and free from importation of horses, usually escape. Thus glanders is very rare in Iceland and in the Faroe islands. In Australia, Tasmania and New Zealand it ts reported to be unknown. Etiology. Bacteriwm mallei, the specific cause of glanders, was discovered and isolated in pure culture almost at the same time (1882) by Loeffler, Schitz, Isrzel, Bouchard, Charrin, Weichselbaum, Kauz- feld and Kitt. It is found in the recent nodules, in the discharge from the nostrils, pus from the specific ulcers, and occasionally in the blood of animals affected with acute glanders. Morphologically it is a small organism with rounded or pointed ends. It varies in breadth from 0.254 to 0.44 and from 1.54. to 3¥. in length. It is usually single but pairs and long filaments, especially on potato cultures, are not rare. It frequently breaks into short, almost coccus-like elements. Galli-Valerio found great variations in its morphology when grown under certain different conditions. Branching forms were numerous. It stains with some difficulty. Of the aniline dyes the best results are obtained with the aqueous solutions, when they are made feebly alkaline. It is decolorized by Gram’s method. It grows well, but slowly, at the body temperature on acid-glycerin agar, in acid-glycerin bouillon, on blood-serum and on potato. Of the test animals guinea pigs and field mice are the most suscepti- ble. In guinea pigs, subcutaneous injections are followed in four or five days by swelling at the point of inoculation and sloughing of the skin, which are followed by the formation of a chronic, purulent ulcer. The lymphatic glands become inflamed and symptoms of general infection develop in from two to four weeks; the glands suppurate and in males the testicles are involved. A purulent inflammation of the joints may occur. The formation of the specific ulcers upon the nasal mucous membrane, which forms one of the characteristics of the disease in the horse, rarely occurs in the guinea pig as a result of inoculation. The disease is often prolonged for several weeks or months. Guinea pigs succumb usually in from eight to ten days when injected into the peritoneal cavity with a virulent culture. In males, the testicles are invariably affected. The inoculation of male guinea pigs and diagnosis of glanders by the orchitis that follows is 112 GLANDERS known as the Strauss method. Hallopian and Bureau observed an orchitis following the inoculation of pus from a case of human mycosis into the peritoneal cavity of a guinea pig. Nocard recorded nineteen cases of a slightly contagious, farcy-like lymphangitis in horses due to a bacterium which produced an orchitis when inoculated into guinea pigs but which was different from Bact. mallet both in its cultural characteristics and its reaction to the Gram stain (Benson). The inoculation of the guinea pig in making a diagnosis can be consid- Fic. 9. THE SO-CALLED “GLANDERS EXPRESSION.” ered as only one factor. The bacterium must. be obtained from the lesions and identified. Infection takes place through the digestive tract. Experimentally it has been induced through the respiratory organs. The period of incubation is not definitely known. It varies from a few to many days, depending upon the method of infection and the virulence of the organisms, as well as the resistance of the animal. Symptoms. Two forms of glanders have been recognized, namely, acute and chronic. Acute glanders. Acute glanders is common in the ass and mule, but Jess frequent in the horse. After a short period of incubation the animal has a chill, elevation of temperature, a profuse muco-purulent, GLANDERS 113 sticky discharge, sometimes mixed with blood, from the nose. Par- ticles of food arrested in the pharynx occasionally appear in the nasal discharge. If unilateral the margin of the nostril swells, the mucosa is dark red, infiltrated, marked with pea-like, yellowish elevations with red areolee, which in a few days become eroded, thus forming spreading ulcers. The submaxillary lymphatic glands on the affected side become enlarged. There may, however, be a uniform swelling of the intermaxillary space. The course is rapid and death may occur in from the sixth to the fifteenth day. The acute form rarely if ever becomes chronic. Chronic glanders. In the horse, this form of the disease may begin with a chill but usually the onset is very insidious. There may be a muco-purulent, sticky discharge, sometimes streaked with blood, from one or both nostrils. There may be intermittent or continued lame- ness, arthritis, edema of a limb, swelling of a testicle, cough, or epis- taxis. There is usually a nodular but comparatively painless swelling of the submaxillary lymph gland on the affected side. On palpation the swelling imparts a sensation suggestive of a number of peas. They are adherent to the adjacent structures. The nasal mucosa is congested, of a dark reddish color and sprinkled with superficial or deep ulcers either clean or covered with crusts. Rarely the submaxillary glands only are apparently diseased. In other cases, there is only a cough, the lesions being confined to the lungs. Occasionally, the lesions are restricted to one or both testicles, the spleen, or other internal organ. Objective symptoms may or may not be present. Chronic glanders may terminate in the acute form. In chronic, cutaneous glanders, with or without edema of the limbs, there may be one or many nodules on the fetlock, or elsewhere on the line of the lymphatic vessels, with induration of the lymphatics extending from it. The nodules may be suppurating and discharging, or they may be closed. The period of incubation and duration found after a lecture on morbid anatomy. Morbid anatomy. In chronic glanders the most frequent locations of the lesions are on the respiratory mucous membrane, in the lungs, lymph glands and skin. M’Fadyean states that he has never seen a case of glanders in which the Jungs were not affected if any lesions were found. Other organs are more rarely invaded. The mucous membrane of the upper respiratory passages is the usual seat of the lesions. Glanders occurs in two forms, (1) as circumscribed nodules with the formation of ulcers and cicatrices; and (2) as diffuse or infiltrated lesions. 114 GLANDERS In nodular glanders, which is the common form, the lesions are most frequently situated on the upper portion of the nasal septum and in the cavities of the turbinated bones. The affection begins with the appearance of nodules vary- ing in size from a grain of sand to a millet seed. They are more or less translucent, of a roundish or oval shape, and of adirty gray or grayish-red color. The nod- ules, which may attain to the maximum size of a pea, project somewhat above the surface of the mucous membrane. They are surrounded by a reddish ring. Some of them are isolated and others are arranged in groups. Microscopically they consist of a large number of lymphoid cells, which disintegrate in the centre of the nodule. In consequence of the central fatty and purulent degeneration, the nodules become yellowish in color, discharge and form ulcers. These ulcers are sometimes superficial, sometimes deep, lenticular or crateriform, surrounded by a hard, indurated edge, and frequently becoming confluent, with irregularly ser- rated and eroded edges. They are sometimes covered with a brownish crust. The ulcers may increase in area or in depth and may even involve the underlying cartilage or bone, causing per- foration of the septum nasi, and distensions of the maxillary or exostoses on the turbinated bones. The shallow lenticular ulcers may heal without Jeaving any visible Fic. 10. NASAL SEPTUM SHOWING ULCERS. GLANDERS 115 changes; but the deeper ones, after granulating, leave a radiating, star-shaped cicatrix which is either smooth or horny, and which, according to the shape of the ulcer, may be of an irregular or oblong Fie. 11. Scar TISSUE FOLLOWING GLANDERS ULCERS, NASAL SEPTUM HORSE. (AFTER JOEST.) ae amare a I - Fig. 12. ScaRS FROM NON-GLANDERED LESIONS ON NASAL SEPTUM HORSE. (AFTER JOEST.) form. The nasal septum is frequently covered with these scars. The ulcers and cicatrices are sometimes found in the maxillary and frontal sinuses, in the guttural pouches and in the eustachian tubes. They may also occur in the larynx, especially in the region of the vocal chords. Inthe trachea and even in the bronchi, particularly on the anterior surface, numerous long, oval ulcers or long, pointed, 116 GLANDERS serrated scars are occasionally found. In addition to the ulcers, a catarrhal inflammation of the mucous membrane is very apt to be present. , Diffuse glanders manifests itself as a diffuse catarrh of the mucous membrane of the nasal and neighboring cavities, with superficial ulceration, thrombosis of the veins, inflammatory in- filtration of the submucosa, considerable thickening of the mu- cous membrane and the formation of a peculiar, radiating cicatrix. Both the nodular and infiltrated forms are found in the lungs. In the nodular form, the lungs contain nodules* varying in size from a millet seed to a pea. They are gray by transmitted light, glassy and pearl gray by reflected light, and are surrounded by a con- gested or a hemorrhagic ring. The center of the nodule shows a pale » Fie. 13. GLANDERS ULCERS IN THE TRACHEA (A) PERFORATION. (AFTER JOEST.) yellow point in consequence of caseation and disintegration of the innermost cells. These nodules are of different sizes, of varying numbers, and of different ages. The formation of a capsule by a con- nective tissue membrane is induced by a reactive inflammation in the tissue surrounding the nodule. The nodules may be of an embolic origin, situated principally in the periphery of the lung, their structure being the same as that of the nodules in the nasal mucosa. Some- times the lung nodules represent lobular pneumonic foci, in which the alveoli are filled with red and white blood corpuscles and with des- *Nocard showed that when glandered horses are treated with mallein, a certain pro- portion of them recover, in which case nodules that are present in the lungs cease to contain living bacteria, a fact he has fully proved by inoculation. On postmortem examination the nodules may be readily felt by passing the hand with firm pressure over the surface of the lung, which, when badly diseased, will feel like a bag full of shot or peas. GLANDERS 117 quamated epithelium of the lungs. Central disintegration occurs very early. These areas are surrounded by a membrane resulting from a reactive inflammation which manifests itself and out of which a connective tissue capsule develops later on. There are two theories Fic. 14. LuNG oF HORSE SHOWING SMALL AND LARGER GLANDERS NODULES. concerning the structure of the early nodules. One is, that the first cells are epithelial in nature, thus closely resembling a tubercle. The other is that the first stage of the nodules consists of air cells filled with leucocytes. 118 GLANDERS M’Fadyean has called attention to the structure of the lung nodules, in which he finds a central part composed of leucocytes that have filled the air spaces, the walls of which have disappeared as if by liquefaction. This is surrounded by a zone of epithelioid cells. A third zone surrounds this, in which the walls of the air vesicles are e Fic. 15. (a) Mass or FIBROUS TISSUE SURROUNDING QUITE LARGE BRONCHI IN GLANDERED LUNG; (b) GLANDERS NODULE. recognizable. The walls are thickened. The fourth zone is com- posed of air vesicles filled with a fibrinous exudate, which entangles a few leucocytes. Frequently the exudate is free from red blood corpuscles, but at times it contains much blood. In older nodules the third and outermost zone is composed of cirrhotic lung tissue, in which can be distinguished the remains of the air cells. This zone passes gradually into the normal tissue. In the last stage the central GLANDERS 119 Fic. 16. LarGB GLANDERS NODULE UNDERGOING ORGANIZATION (Its DEVELOPMENT OF FIBROUS TISSUE). Fic. 17. GuanpEerRs NopULE. Low MAGNIFICATION 120 GLANDERS area shrinks and becomes calcified, while the other zones become con- verted into a distinct fibrous capsule. Other observers have not reported the calcification. It has not occurred in the writer’s observation. The cell necrosis in glanders has been designated by Fic. 18. A porTION OF A GLANDERS NODULE (FIG. 17) EXTENDING FROM THE NECROTIC CENTER TO THE NORMAL TISSUE. X 380. Unna as chromatolasis which consists in the disintegration of the nucleus before the destruction of the cell body and the retention of the staining property of the broken, nuclear chromatin. This gives the dark color in the central part of a stained nodule. Besides these nodules, there are often chronic bronchitis, peribron- chitis, parabronchitis, atelectasis, inflammation of the tissue of the Jung and less frequently circumscribed or exudative pleuritis. GLANDERS 121 Infiltrated glanders of the lungs forms tumors from the size of a walnut to that of a child’s head, consisting of a diffuse glanderous infiltration of the alveoli and of the interstitial connective tissue. Frequently on section the infiltrated parts of the lungs resemble very closely a soft sarcoma. They are of a dirty white color, of a gelatin- ous, juicy consistency and irregular in shape. They may either become indurated so as to form hard, connective tissue-like new growths (fibroma-like tumors of glanders, according to Gerlach), or they may become gangrenous. At times there appear masses of connective tissue of varying size at the borders of which glanders bacteria are found. In nodular and in infiltrated glanders of the lungs, the bronchial glands and frequently the mediastinal glands become enlarged, indurated and studded with smal! foci of cell infiltration. In glanders of the skin (farcy) the nodules are found in the papillary layer, in the cutis and in the subcutaneous and superficial intermuscu- lar tissue. The cutaneous a nodules vary in size from a hemp seed up toa pea. They suppurate rapidly and form small ulcers. The nodules in the subcutis are inflammatory (metastatic) tumors from the size of a pea to that of a hen’s egg. They change into large abscesses and discharge ex- ternally. In the region of the nodules the lymphatic vessels are inflamed, swollen, and fre- quently resemble a rosary or knotted cord. Ulcers often develop from these secondary nodes. The neighboring lymph glands are at first swollen and soft, but later : they become indurated by the Fic. 19. Smcrion oF A GLANDERS i S NODULE IN THE LUNG OF A HORSE: growth of connective tissue (a) NECROTIC CENTER, (¢) ZONE OF and studded with dirty white GIANT CELLS, (b) CAPSULE SURROUND- : ING THE NODULE (Schutz). nodules about as large as a pin ( ) head, or with yellow foci of caseation. The capsule around the 122 GLANDERS lymph glands becomes infiltrated with small cells and subsequently thickened. In rare cases secondary chronic farcy occurs. It is marked by a large, diffuse new growth of connective tissue with nodular thickening of the skin. This condition is termed glander- ous elephantiasis or pachyderma. It chiefly affects the limbs and head. Of the abdominal organs, the spleen is most frequently attacked. It then contains embolic nodules, which vary in size and either suppurate or become calcareous. Similar nodules occur, though not so often, in the liver, kidneys, testicles, brain, muscles, heart and bones. In the bones, the lesions consist of a cellular infiltration of the medulla and purulent breaking down of the osseous tissue. Ulcers are Fic. 20. GLANDERS NODULES IN Fic. 21. GLANDERS NODULE IN SPLEEN LYMPH GLAND DISCHARGING INTO OF HORSE. NATURAL SIZE. BRONCHUS. (a) BRONCHI, (b) LYMPH GLAND, (¢) OPENING INTO BRONCHUS. very rare on the mucous membranes of the eyes, stomach and vagina. The blood shows signs of slight leucocytosis. The specific bacteria are found in the blood only in cases of acute general infection. The anatomical changes in acute glanders consist chiefly in a dis- integration of the respiratory mucous membrane, in a serous infiltra- tion of the submucosa, subcutis, and intermuscular tissue, with in- flammation and suppuration of the lymph vessels and glands. There are also metastatic formations in the skin and lungs. The nasal mucous membranes are covered with rapidly spreading ulcers with considerable infiltration into the submucosa. The mucous membrane of the larynx and pharynx may be swollen and covered with ulcers. The lungs are studded with purulent metastatic foci or fresh nodules. The skin is excessively swollen and covered with glanderous nodes. Sometimes diffuse gangrene of the skin occurs. GLANDERS 123 Glanders in man. The symptoms of glanders in man are of much importance to the veterinarian. Although the susceptibility to the disease is usually not very great, cases of human glanders unfor- tunately occur, especially among veterinary surgeons and those hav- ing the care of horses. Human glanders is reported to be quite com- mon in Russia. Robins has reported 156 cases collected from the Fic. 22. SKIN GLANDERS (FARCY). literature. The parts usually first affected are the hands, nasal mucous membrane, lips and conjunctiva. After a period of incuba- tion of from three to five days the infected part becomes swollen and painful, with subsequent inflammation of the lymph vessels and swelling of the glands. Fever is often the first symptom, and it is nearly always followed by a nasal discharge, ulcers on the nasal mucous membrane, pustules and abscesses in the skin, ulcers in the oral cavity, larynx, and conjunctiva, articular swellings, and grave general disturbances. Sometimes there is intense gastro-intestinal trouble. Nodules occur in the lungs in some cases. Asa rule, death 124 GLANDERS takes place in from two to four weeks. In other instances, the disease becomes chronic, lasting for months or years. Bact. mallet has been found in the blood in cases of acute glanders. The positive diagnosis depends on the possibility of infection having taken place, on inoculation in guinea pigs, the proof of the presence of Bact. mallei or positive results with sera tests. Treatment is usually of no avail. The only hopeful cases are those that are purely local in their manifestation. A few of these are reported to have been cured by applying deep cauterization. Diagnosis. Glanders is to be diagnosed by the symptoms, lesions, cause and specific reactions of which there are several. In somewhat “typical” and advanced cases the diagnosis may be very accurately made from the manifestations. The positive diagnosis, however, must be made from the identification of Bact. mallet, or from one or more of the specific tests. In the dead animal the histology of the lesions may reveal the nature of the disease. Lesions. The lesions exhibited in a living animal are not suffi- ciently characteristic to positively identify the disease in very many, if in any, cases. The histological structure of the nodules may, how- ever, enable one to do so. Bacteriological examinations. The examination of the lesions for Bact. mallet can be made by cultures or guinea pig inoculation. In case of nodules in internal organs cultures on acid-glycerine agar or bouillon or on potato are quite satisfactory. Animal inoculation. Male guinea pigs should be used. The material for inoculation usually consists of the nasal discharge from the suspected glandered horse, bits of scrapings from the ulcers, or pieces of other diseased tissue may be injected subcutaneously or into the abdominal cavity. The first symptom of glanders noticed is usually orchitis. The lymphatic glands in the groin are also enlarged. After the orchitis becomes well advanced, the guinea pig may be chloroformed and examined. Pure cultures of the specific organism can be obtained in most cases from the suppurating foci in the testicle. The spleen is usually enlarged and sprinkled with grayish nodules. Other organs may be involved. The diagnosis by the inoculation of a male guinea pig is known as the Strauss method. It is important to note that the orchitis alone is not sufficient to make a positive diag- nosis but the specific organism must be found and identified. Mallein. Mallein is prepared in the same way as tuberculin. It was used by Kaling and Helman independently as a diagnostic agent. GLANDERS 125 It consists of the glycerinated bouillon in which the glanders bacteria have grown and in which are the products resulting from their multiplication. It has a somewhat fetid odor. Mallein is applied in two ways, namely, subcutaneously and on’ the conjunctiva. The latter is called the ophthalmic method. Subcutaneous method. In applying mallein the horse is injected usually in the neck with from 0.5 to 2 cc. of mallein, the quantity depending upon the degree of concentration. If a concentrated mallein is used it should be diluted with a 1 per cent. carbolic acid solution to at least 2. cc. The reaction is as follows. In a few hours there forms at the place of injection a hot, inflammatory swelling. It is very painful and in case of glanders quite large. From all sides of the swelling there may radiate wavy lines consisting of swollen lymphatics, hot and painful when touched, extending toward the adjoining glands. When the mallein injection is made aseptically, this swelling never suppurates, but it may increase in size during a period of from 24 to 36 hours and persists for several days, when it gradually diminishes and finally disappears at the end of eight or ten days. With the appearance of the local swelling the patient becomes dull and dejected, the eyes have an anxious expression, the coat is lusterless, the flanks contracted, the respiration hurried and the appetite isimpaired. Frequent shudders are observed to pass through the muscles of the fore legs and sometimes the trunk is subject to violent convulsive movements. The most active and fractious horses become listless and indifferent to their surroundings. These general phenomena constitute what the French call the “organic reaction,” but they are not always so clearly marked. Differences in their intensity are observed but they are never completely absent. The temperature reaction seldom fails to show itself. In about eight hours after the injection the temperature of a glandered horse gradually rise 1.5°, 2° or 2.5° F., and even more above the normal. The rise in temperature usually attains its maximum between the tenth and twelfth hour, occasionally not till the fifteenth, and more rarely not until about the eighteenth hour. An important fact to note is that the reaction called forth in glandered horses by the injec- tion of mallein persists for from 24 to 48 hours and in some cases the temperature remains above the normal for an even longer time. In practice it is advisable to take the temperature of the suspected animals two or three times before the injection of the mallein, and every two hours, beginning at the eighth and going to the twentieth 126 GLANDERS hour after the injection. It is often sufficient for diagnostic purposes to take the temperature but four times, viz., at 9, 12, 15, and 18 hours after the injection, but a longer observation would be more reliable. aa MAU Am [AM |AALIAM. | M. | PM.) PAM) PM. eem|| 4 | 6 | & | so |7e | 2 | 4 8 Bq LQ Ny NX >» S S ite » iB = BS DN BK PW q Ns RAR 37 NK N ~~ qd MN LQ2' Vieeesil ewe and x OF Es A / “UVF vaaYal re Oe Fic. 23. MALLEIN REACTION. TEMPERATURE CURVES OF SIX HORSES FOR 24 HOURS AFTER INJECTING MALLEIN. THE HORSES WERE IN ONE STABLE FROM WHICH A WELL DEVELOPED CASE OF GLANDERS HAD BEEN REMOVED. In healthy horses the injection of mallein, even in a much larger dose, produces no effect on the temperature or the general condition of the animal. There is produced, however, at the point of injection, GLANDERS 127 a small edematous swelling, somewhat hot and painful to the touch, but the edema instead of increasing, diminishes rapidly and disappears in Jess than 24 hours. The reaction called forth by the injection of mallein in a glandered animal is quite specific. When it occurs one is enabled to state at once and with certainty that glanders exists, although the lesions may be quite minute or obscure. When the reaction does not take place it is generally considered that the animal tested is not glandered, although the physical examination may suggest it. Notwithstanding the specific action of mallein, its administration can give really useful indications “‘only when, and as far as, we can remove the causes of error that have been pointed out by experience.’ For example, it would be imprudent to use mallein in case of animals already suffering with an abnormally high or low temperature. The further precau- tions should be taken that the animals subjected to the test be removed as far as possible from atmospheric variations and the influence of strong sunlight, fog, rain and currents of air. If it be true that the majority of horses are not susceptible or slightly so, to these influences, there are still some that are affected by them. A sudden rise and fall of temperature due to other causes must be differentiated from a mallein reaction. Ophthalmic use of mallein.* Schniirer in Vienna and Fréhner in Berlin recommend this method of using mallein. The mallein is applied to the eye with a camel’s hair brush in the following way: The eyelids are opened with the index finger and the thumb, as is customary when examining the conjunctiva of the eye. Then the camel’s-hair brush, which has been submerged in the mallein, is drawn once forward and again backward over the eye. Only one eye is used, the other serving as a control. Immediately after the application of the mallein to the eye in most of the animals Jacrima- tion, increased reddening, and twinkling of the eye appear; these primary reactions are not specific and disappear in the following few hours. The specific reaction commences as a rule 5 or 6 hours after the application of the test and lasts from 36 to 48 hours, occasionally, even longer. It consists in a suppurative conjunctivitis, with redden- ing, swelling, and suppurative secretions. Of these signs only a sup- *The ophthalmic use of mallein was first reported in this country by Dr. C. J. Mar- shall (Proceedings of the A. V. M.A. 1912). At the same time Moore and Fitch were using this method for the diagnosis of glandersm New York City (Report New York State Veterinary College, 1911-12). 128 GLANDERS purative secretion should be taken into consideration. The results are interpreted as follows: (1) The reaction is positive if a suppura- tive secretion is observed in varying quantities. If the secretion is present in only a small quantity, it is principally visible at the inner canthus of the eye. (2) The reaction is negative in the absence of any secretion. (3) The reaction is doubtful when there is present a slimy secretion or lacrimation after 24 hours. The diagnosis should be made not earlier than 12 hours and not later than 24 hours after the application of the test. The examination should be made in a good light. A positive result indicates with certainty the presence of glanders; negative results, however, should not eliminate the possibility of the presence of the disease, and only a repeated negative test after three weeks excludes suspicion of the disease. Generally the positive oph- thalmo reactions are not ac- companied by fever or systemic disturbances. Occasionally, however, affected horses are hypersensitive, so that often a trace of mallein which enters the circulation produces fever. Accordingly, it is advisable to accompany the ophthalmo reac- tion with temperature measure- ments. For this purpose the temperature should be taken at least twice, the first time when the mallein is applied and the second time when it is judged. In a doubtful eye reaction where there is a rising temperature of over 101.5° F., the test should be considered positive if the animal had a normal temperature at the time the mallein was applied. The ophthalmic test is used officially in Austria. They employ the Pasteur “Mallin brute,” 0.75 cc. being used on 10 horses. In this Fic. 24. Ey, roLLOWING OPHTHALMIC USE OF MALLEIN. GLANDERS 129 country both the concentrated mallein and a 5 to 7% solution of the precipitated mallein (Mallein Siccum Foth) are used. The ordinary mallein used for subcutaneous test cannot be relied upon. This test should not be made in the presence of a conjunctivitis. It is also important, where there is a reaction, that the purulent dis- charge be not wiped out of the eye thereby leaving one in doubt. Like the subcutaneous method, there is no definite relation between the extent of the reaction and the amount of pathological changes that exist. The cutaneous and intradermal application of mallein do not give uniformly satisfactory results. The agglutination method or serum diagnosis. Rabieaux found that the difference which exists between the agglutinating power of a serum from a glandered and from a healthy horse may be used as the basis of a method for diagnosing glanders. He collected the serum as pure as possible, diluted it with sterile, distilled water to 1 in 10, or to 1 in 500. The diluted serum was then mixed in a small sterile tube with an equal volume of a 24 to 72 hour culture of Bact. mallei in peptonized bouillon (without glycerin). The mixture was placed in an incubator at a temperature of 35° to 37° C. and examined at variable times under the microscope. In dilutions of from 1 in 10 to 1 in 50 the agglutination occurred in 20 minutes to 3 hours. In serum of a non-glandered horse from 2 to 6 hours were required to produce the agglutination. In weaker dilutions the differences were more marked. The development of the method can be followed from the writings of M’Fadyean, Bourget and Méry, Arpad, Fedorowsky, Reinecke, Bonome, Schiitz and Miessner, Schntirer and Moore, Taylor and Giltner. The method consists in the preparation of a test fluid from a suitable culture of Bact. mallet to which is added the diluted serum. The “‘test-fluid” is prepared by washing the growth from a 72 hour acid-agar culture by the aid of a sterile wire loop into distilled water containing 0.85 per cent. sodium chloride and 0.5 per cent. carbolic acid crystals. This suspension is then placed in a thermostat at 60° C for two hours, which kills the bacteria. Three cubic centimeters of the “‘test-fluid”’ are placed in each of several small test-tubes. Witha sterile pipette, the diluted serum is added to the tubes of test-fluid and thoroughly mixed. In making the different dilutions, the amount of diluted serum to be used is readily ascertained by the following table: : 130 GLANDERS Dilution of Serum Panount: satan Diluted | Amount cf Test Fluid Dilution 1-40 1.2 ce. | 3 ee. 1-100 1-40 0.6 3 1-200 1-40 0.405 3 1-300 1-40 0.3 3 1-400 1-40 0.24 3 1-500 1-40 0.195 | 3 1-600 1-40 0.15 | 3 1-800 1-40 0.12 3 1-1,000 1-40 0.105 3 1-1,200 1-40 0.09 3 1-1,500 1-40 0.06 3 12,000 1-40 0.03 3 1-4,000 1-40 0.015 3 1-8,000 Where dilutions greater than 1-1000 are made, a serum diluted 1-80 may be used to better advantage, unless the pipette employed is very finely graduated. In this case the amount of diluted serum for a certain dilution must be double that indicated in the table. The mixture thus prepared is placed in an incubator at 37° C. for 24-30 hours. A temperature higher than 37° C. interferes with the agglutination. The reaction consists of a layer of the agglutinated bacteria cover- ing the entire convexity at the bottom of the tube. This film-like sediment may become so dense that it rolls in at the periphery. The supernatant fluid becomes clear in the lower dilutions, but in the higher ones the clarification may not be complete, showing that all the bacteria have not become agglutinated. This is further evidenced by the fact that the layer is less dense in the higher dilutions. The reaction may begin in six hours, but cannot be considered complete until 24 to 36 hours have elapsed. If no reaction appears in 24 hours it cannot be considered negative, as it may occur in from 30 to 40 hours after setting. Often, however, a reaction appears in less than 24 hours. After the agglutination is completed, further standing produced no visible change in the test fluid. A negative result shows a small round concentrated spot of sedi- ment in the center of the convexity at the bottom of the tube, the test fluid remaining apparently unchanged even after several weeks. Animals whose blood serum agglutinates in dilutions of 1-500 are GLANDERS 131 suspicious and a reaction in dilutions of 1-800 or higher indicates an infection with Bact. mallet. As pointed out by Bonome and confirmed by Taylor, there seems to be little or no change produced in the precipitating power of the serum of the blood taken before, during or after the mallein reaction but the agglutinating power as determined microscopically is very much increased during the mallein reaction. This is shown by the appended table: TABLE SHOWING BOTH MACROSCOPICALLY AND MICROSCOPICALLY THE AGGLUTINATION OF DEAD GLANDERS BACTERIA WITH BLOOD SERUM FROM HORSES TAKEN BEFORE, DURING AND AFTER THE MALLEIN REACTION. Blood taken the day previous to Blood taken during the || Blood taken after the tempera- malleination Feb. 21 reaction Feb. 22 ture had sparred to normal No. |Macroscopic | Microscopic || Macroscopic | Microscopic || Macroscopic Microscopic 1 1-800 1-1000 1-800 1-1800 1-800 1-1000 4 1-800 1-1200 1-1000 1-2000 1-1000 1-1500 5 1-1500 1-1600 1-1500 1-2500 1-1000 1-1200 6 1-1200 1-1500 1-—1200 1-2000 1-1800 1-1800 i 1-1000 1-1200 1-1000 1-1800 1-1000 1-1400 8 1-500 1-500 1-800 1-1600 1-500 1-600 9 1-800 1-1000 1-800 1-1400 1-500 1-800 10 1-500 1-600 1-500 1-1000 1-500 1-800 12 1-200 1-300 1-500 1-800 1-500 1-750 13: 1-1000 1-1200 1-500 1-1000 1-800 1-1000 14 1-500 1-600 1-500 1-800 1-500 1-800 15 1-1200 1-1400 1-1200 1-2500 1-1200 1-1800 16 1-1000 1-1200 1-1000 1-2400 1-1000 1-—2200 17 1-500 1-800 1-800 1-1500 1-800 1-1400 18 1-1000 1-1400 1-—1000 1-1800 1-800 1-1500 19 1-1000 1-1000 1-800 1-1200 1-1000 1-1200 20 1-500 1-600 1-500 1-800 1-500 1-800 21 1-500 1-750 1-500 1-1000 1-800 1-1200 22 1-1000 1—1400 1-1000 1-2500 1-800 1-2200 23 1-200 1-200 1-200 1-1000 1-800 1-—1000 2A 1-1000 1-1200 1-1000 1-2200 1-1200 1-2000 WA, 1-1000 1-1200 1-1000 1—2200 1-1200 1-2000 25 1-800 1-800 1-500 1-1200 1-800 1-1000 26 1-1200 1-1500 1-1000 1-—2200 1-1200 12000 Q27 1-500 1-1000 1-1000 1-1800 1-800 1-2000 28 1-500 1-600 1-500 1-1200 1-1000 1-1500 29 1-800 1-1000 1-500 1-1200 1-1200 1-—2000 30 1-1000 1-1200 1-800 1-1800 1-800 1-1600 31 1-500 1-600 1-800 1-1400 1-1000 1-1500 182 GLANDERS The agglutination in higher dilutions with the living organisms as determined microscopically was pointed out by Taylor. A compari- son of the agglutination of the living and killed bacteria with the serum from glandered horses, as shown by the mallein reaction, is given in the appended table: a MACROSCOPIC AND MICROSCOPIC AGGLUTINATION OF BACTERIUM MALLEI WITH HORSE SERUM BY THE USE OF KILLED AND LIVING CULTURES. Macroscopic. Microscopic. Microscopic. Number of Horse | Dead bacteria, 24 hours | Dead bacteria, 12 hours | Live bacteria, 12 hours at 37°C. Bt a7? C. at 37°C, a 1-8000 1-12000 1-30000 g 1-2000 1-3000 1-12000 3 1-800 1-1000 1-10000 A 1-1600 1-1800 1-6000 5 1-500 1-600 1-5000 6 1-1000 1-1250 1-25000 7 1-800 1-1000 1-8000 8 1-800 1-1200 1-12000 9 1-1600 1-1800 1-24000 10 1-500 1-800 1-7500 The method as pointed out by Schiitz and Miessner is a macro- scopic one. It depends upon the precipitation of the agglutinated masses of bacteria. Normal horse’s serum agglutinates glanders organisms in high dilutions as determined microscopically. This, however, does not appear to be of diagnostic value. Complement fixation. This is strictly a laboratory method and cannot be applied in the field. It requires the collecting of the blood from the suspected animals and sending it to the laboratory as quickly as possible. The method is fully described by Mohler and Eichhorn in Bulletin 136 of the Bureau of Animal Industry. Schiitz and Schubert, after applying this method and comparing it with others, recommend it as the most accurate means for diagnosing glanders. It is widely used in this country usually in conjunction with one or more of the other tests, especially the ophthalmic use of mallein. The test requires five different substances: (1) washed red blood corpuscles of a sheep, (2) hemolytic amboceptor, (3) complement, (4) antigen (bacterial extract), (5) serum from the blood of the suspected animal to be tested. Red blood cells. The washed red blood corpuscles of a sheep are obtained by bleeding a vigorous sheep from the jugular vein under antiseptic precautions. The blood is preferably collected in a sterile flask containing a few glass beads. The blood is shaken, defibrinated and filtered through sterile gauze into a glass tube. The tube is then filled GLANDERS 133 with physiological salt solution and the mixture centrifuged*. The supernatant fluid is poured off and the process repeated until all the serum is removed from the corpuscles. Enough salt solution is added to the sedimented corpuscles to make the volume of blood equal the original amount filtered into the tube.’ Hemolytic amboceptor. The hemolytic amboceptor is obtained from the serum of a rabbit which has been immunized to the washed red blood corpuscles of a sheep. A strong vigorous rabbit is selected and injected with the washed red corpuscles. The injections are preferably made intraperitoneally and at intervals of 4-5 days. Two ce., 4 ce., 8 cc., and 12 ce. of the corpuscles are respectively used for each injection. In seven or eight days after the last injection a small amount of blood is taken from the rabbit (by bleeding from an ear vein) and the serum titrated to determine whether its hemolytic action is sufficient; that is, whether it will readily dissolve the hemoglobin from the corpuscles of a sheep. If it is found satisfactory the animal is bled from the carotid and the serum collected and stored in small bottles. It is preferable not to put more than 2 cc. in each bottle. The serum may be preserved by adding 0.5 per cent. of carbolic acid in a 5 per cent. dilution. If the carbolized serum is not used for three days after adding the carbolic acid it will not require inactivation (heating at 56° C. for Y% hour). Noguchi prefers adding 2 drops of chloroform to each 2 cc. of hemolytic serum and in this case in order to get the best results the serum should be inactivated before using. Serum preserved in this way can be kept in the ice-box for from three months to a year. It must, however, be titrated at intervals of a few weeks as the titre may change. Titration of hemolytic rabbit serum. Dilutions of the hemolytic serum are made in eight test tubes, according to the following table. It is best to use pipettes to measure the required amounts. TABLE No. I DILUTIONS OF HEMOLYTIC AMBOCEPTOR (RABBIT SERUM). Tub NaCl Gi Ne a Selution scale : Giuten sce 1 9 ce. lee. 1-10 | This isa 1-10 basic dilution. 2 9cc. | 1 cc. of 1-10 1-100 dilution 2 8 ce. 2 cc. of 1-100 1-500 dil. 4 9 ce. 1 ce. of 1-100 1-1000 dil. 5 lee. | 2ec. of 1-1000 1-1500 dil. 6 lee. | 1lec.of 1-1000 | 1-2000 dil. 7 3 ce. 1 ce. of 1-1000 1-4000 dil. 8 8cc. | 2cc. of 1-1000 1-5000 dil. ‘Fer hemolytic work 0.85 per cent. (Ehrlich) to 0.9 per cent. (Madsen) salt solution is universally employed. We prefer the latter concentration. 134 GLANDERS The titration proper is then made in the following manner: Eight additional test tubes are each filled with 2.5 cc. of salt solution to which is then added the hemoly- tic serum (amboceptor) in quantities of 1 cc. of the different dilutions (Table No. I) to each tube. Afterwards the complement of the guinea pig serum is added in quantities of 0.5 ce. of a 10% dilution to each tube, and finally 1 cc. of a five per cent. suspension of washed sheep corpuscles in salt solution is placed in each tube. Besides these eight tubes there are also three control tubes, one to show that the complement alone will not produce hemolysis (without the amboceptor), the second that the amboceptor alone without the complement will not produce hemolysis, and the third that the salt solution alone will not produce hemolysis. Thus in the first control tube we add 3.5 cc. of salt solution, 0.5 cc. of complement, and 1 cc. suspension of sheep corpuscles. In the second control tube we add 8 ce. salt solution, 1 cc. of the 1-100 dilution of the amboceptor and 1 cc. suspension of sheep corpuscles. In the third control tube we add 4 ce. salt solution and 1 cc. sheep corpuscles. A test tube rack with two rows of holes is very convenient for holding the tubes for these tests. TABLE No. II TITRATION OF RABBIT SERUM (HEMOLYTIC AMBOCEPTOR). Tube | (a) NaCl Amboceptor (b) Com | (c) Blood Remarks No. solution plement jcorpuscles 1 2.5 ec. | 1 cc. of 1-10 dil. 0.5 ce. lee. 2.5 ec. | 1ce. of 1-100 dil. | 0.5 ce. l ce. 2 3 2.5 ec. | lec. of 1-500 dil. | 0.5 ec. | Lee. 4 2.5 cc. | Lec. of 1-1000 dil.| 0.5 cc. lee. 5 2.5 ec. | 1 ec. of 1-1500 dil. 0.5 ce. lee. 6 2.5 cc. | 1ce. of 1- 2000 dil.} 0.5 ec. lee. 7 2.5 ec. | lec. of 1-4000 dil.| 0.5 ce. | 1 ce. 8 2.5 ce. | 1 ec. of 1-5000 dil.| 0.5 cc. | 1 ce. controls Complement control 9 3.5 ce. 0.5 ce. | lce. {no hemolysis should occur) .. Amboceptor control (no 10 3.0 ce. | 1 ec. of 1-100 dil. lee. hemolysis should occur) 11 4.0 ce. lee. {Salt solution control (no hemolysis should occur) a. 0.9% NaCl solution. ‘ b. 0.5 cc. of a 10% solution of the ccmplement. c. 59% suspension of washed sheep corpuscles in salt solution. As can be seen from Table II the final volume in each tube is always uniformly 5 ce. Thus the different amounts of the blood derivatives used are always made up to.5 ce. by adding salt solution. GLANDERS 135 After adding the substances to the test tubes in the order given, the tubes are well shaken, placed in the test tube rack, and put in an incubator at 37° C. for two hours. Then the tubes are removed from the incubator and the results read.* The highest dilution in which complete hemolysis has taken place represents the titre of the hemolytic amboceptor. Thus if complete hemolysis has taken place up to and including the tube where the dilution of the rabbit serum was 1-2000 (Tube No. 6, Table ITI), the hemolytic titre of this serum is represented by 1-2000. This dilution, however, is not used in the glanders test; but rather its double strength, which would be 1-1000. The titre of the hemolytic amboceptor for use in the diagnosis of glanders should not be less than 1-1000 and therefore if the rabbit serum should prove to be of a lower titre it should be discarded. It often happens that some rabbits are found to be unsuited for the production of amboceptor and the titre of their serum can not be raised high enough even after repeated inoculations with washed sheep corpuscles. It is advisable to preserve the hemolytic amboceptor in small vials containing 1 to 2 ce. of the rabbit serum, and seal the corks with paraffin or sealing wax. Complement. The complement which is contained in the blood serum of a normal guinea pig is employed. The complement should be titrated to determine its efficiency to act with the hemolytic amboceptor. Titration of complement. By titration of the complement there is aimed to be established a complement unit which is the smallest quantity of complement necessary TABLE No. III TITRATION OF COMPLEMENT. Complement Tub NaCl x, Amboeep- No. | edlanoe (a) gone © tor (c) Blood corpuscles (d) 1 2.5 ce. 0.5 ce. lee. lee. 2 2.6 ce. 0.4 ce. lee. lee. 3 2.7 ec. 0.3 ce. 1 ee. lee. 4 2.8 cc. 0.2 ce. lee. 1 ee. 5 2.9 cc. 0.1 cc. lee. l ce. controls 6 3.5 ce. 0.5 ce. 1 cc. complement control (no hemolysis should occur) 7 3.0 ce. lee. | 1 ec. amboceptor control (no hemolysis should occur) 8 4.0 cc. 1 ce. salt solution control (no hemolysis shculd occur) a. 0.9% NaCl solution. as b. Guinea pig serum in diminishing quantities. __ . : ce. Of previously titrated hemolytic serum, double dissolving quantity. d. 5% suspension of washed sheep blood corpuscles in salt solution. *If the tubes are placed in a water bath kept at 37° C. the time here may be shortened to one-half hour. 136 GLANDERS to produce complete hemolysis in the presence of one amboceptor unit and a suspension of sheep corpuscles.* The serum from each guinea pig should be titrated in the follow- ing manner before it is used. A basic dilution of 1-10 is made of the complement by adding 0.3 cc. of the complement to 2.7 cc. of salt solution in a test tube. From this dilution certain definite amounts are added to each of five other tubes according to Table III. To the complement thus distributed to each tube, is added 1 ce. of hemolytic amboceptor of which the titre has already been determined{ and likewise 1 cc. of a 5% solution of sheep blood corpuscles. The amount of fluid in each tube is made up to 5 ce. by the addition of salt solution. Three controls are used. The first control tube contains 3.5 ce. salt solution, 0.5 cc. of the 10% basic dilution of complement and 1 cc. of a suspension of sheep corpuscles. In the second control 3 ce. salt solution, 1 cc. of the amboceptor dilution and 1 cc. of the suspension of corpuscles; in the third control 4 cc. of salt solution and 1 ce. of the suspension of sheep’s corpuscles are used. The order of adding each product and the amount of each are given in Table III. After shaking each tube, they are placed in a rack and then in an incubator at 37° C. for two hours.{ After the expiration of this time they are removed and the results noted. The highest dilution of complement in the tube in which the hemolysis is com- plete indicates the titre of the complement. For example, if hemolysis is complete in the tube where 0.3 cc. of the 10% basic dilution of the complement was used (Tube 8, Table III) and the hemolysis is incomplete in the tube in which 0.2 cc. of the same dilution was employed (Tube 4, Table III) then the titre of the complement is 0.3 cc., inasmuch as a 10% basic dilution of the complement was employed. Thus, in the tests using this complement it would be necessary to employ a 3% complement dilution, that is, 3 cc. of the serum of the guinea pig in 97 ce. of salt solution. The amboceptor used in this titration should be inactivated when fresh, or when preserved with chloroform. The carbolized amboceptor may be used without inactiva- tion after it is 3 days old. Antigen. In testing for glanders this consists of an extract of Bact. mallet prepared as follows. Cultures are made on acid glycerin agar and allowed to incubate for 48-72 hours. The growth is then washed off with salt solution. This salt solution suspension of the organisms is kept at 60° C. for four hours in order to kill the bacteria. The sus- pension is then placed in a shaking machine and shook at frequent intervals for four days. It is then centrifuged at high speed (8000-5000 revolutions per minute), the clear supernatant liquid drawn off and ten per cent. of a 5% solution of carbolic acid added. Before use the antigen must be titrated to obtain its anti-complementary action. It would be well also to determine its antigenic and hemolytic properties as well. However, this may be left until the test proper. Titration of the extract or antigen. The titration of the extract is carried out in order to determine the quantity of the extract which no longer prevents hemolysis *The serum is obtained by bleeding a strong vigorous guinea pig by cutting the throat. The pig is first anesthetized and the ventral portions of the neck shaved and disinfected. The blood is collected preferably in a Petri dish. {That is, if the titre was found to be 1-3000 by the previous titration you would add 1 ce. of a 1-1500 dilution, as double the quantity of amboceptor is added which has been stated befcre. ice again the time may be shortened to one-half hour by the use of a water bath at 87°C. GLANDERS 137 (the anti-complementary action). A 1-10 dilution of the extract is first made by adding 1 ce. of the extract to 9 cc. of salt solution. From this 1-10 dilution other dilutions are prepared as follows: TABLE No. IV DILUTIONS OF THE ANTIGEN. No. | sdiition | Amount of Antigen | ie 1 2 ec. lee. of 1-10 dil. 1-30 2 |. 4ee. lee. of -110 dil. 1-50 3 5 ce. lce. of 1-10 dil. 1-60 4 7 ce. lec. of 1-10 dil. 1-80 5 9 ce. lec. of 1-10 dil. 1-100 6 lee. lec. of 1-100 dil. 1-200 7 1% ce. 1 cc. of 1-100 dil. 1-250 8 2 ce. 1ce. of 1-100 dil. 1-300 9 3 ec. lec. of 1-100 dil. 1-400 10 4 ce. lce. of 1-100 dil. 1-500 The titration proper is carried out as follows. Eleven test tubes and three for con- trols are used. To each tube 1 cc. of salt solution is added, then 1 cc. of the complement of the previously determined smallest quantity established by the titration done before. To each tube add 1 ce. of the different dilutions (1-10, 1-30, 1-50, etc.) of the extract as obtained in Table IV. The tubes are shaken, placed in a rack and the rack placed in an incubator at 37° C. for one hour.* The tubes are then removed and 1 ce. of a double quantity of the previously titrated amboceptor is added to each tube, and finally 1 ce. of a 5% suspension of washed sheep blood corpuscles. Three control tubes are used. The first serves for a control to show that the complement alone (without the ambocep- tor) does not produce hemolysis, the second to show that the amboceptor alone does not produce hemolysis and the third that the salt alone does not produce hemolysis. In the first control tube 3 cc. of salt solution, 1 cc. of complement and 1 cc. of blood corpuscles are added. The second tube contains $ ce. of salt solution, 1 cc. of ambocep- tor and 1 cc. of blood corpuscles, while the third contains 4 cc. of salt solution and 1 cc. of suspension of sheep blood corpuscles. The following table (Table V) gives a sum- mary of the foregoing method. Each tube is shaken and all the tubes placed in the incubator at 37° C. for two hours.t Theresultsare then read. The tube in which hemolysis is no longer prevented represents the titre of the extract. In the glanders test, however, one-half that quan- tity is used. For example, if the results of the titration should be that the first tube *May place in water bath at 37° C. for ¥% hour. +Or water bath at 37° C. for one hour. 138 GLANDERS which does nét prevent hemolysis contains a dilution of 1-80 (tube 5, Table V), then a dilution of 1-160 of the extract is used for the glanders test. Antigen which possesses too much anti-complementary substance should not be used; that is, where hemolysis is prevented in as high dilutions as 1-300, 1-400, 1-500, etc. Suspected serum. The suspected glandered horse is bled usually through the jugular and about 20 ce. of the blood is collected in a small bottle. The blood is then allowed to coagulate and the serum to collect. If the blood came from an animal affected with glanders, the serum contains the specific glanders bacteriolytic amboceptor or immune bodies. The amount of these immune bodies in the blood of glandered horses is not TABLE No. V TITRATION OF GLANDERS BACTERIA EXTRACT, Tube | NaCl so- | Comple- Ambocep-|Blood cor No. | lution (a) | ment (b) Extract (c) tor (d) |puscles (e) 1 lee. lec. | 1lec.of 1-10 | lee. 1 ce. 2 lee. lee. | lee.of 1-30 | lce. l ce. 3 lee. lee. | lec.of 1-50 | 1 ce. l ce. 4 lee. lee. | lec.of 1-60 | 1 ce. 1 ee. 5 lce. lee. | 1lec.of 1-80 | Ilce. lee. 6 lce. lee. | 1lec.of 1-100) 1 ce. 1 ce. 7 lee. lee. | lec. of 1-200] 1 cc. lee. 8 lee. lee. | lec. of 1-250} 1 ce. 1 ce. 9 1 ce. lee. | lee. of 1-300} 1 cc. l ce. 10 lee. lec. | lec. of 1-400} 1 ce. lee. 11 lee lee. | 1lec.of 1-500} 1 ce. lee. 12 3 ec. lee controls lec. | Comp. control (no hem- olysis should occur) 18 3 ce. lee. lec. | Ambocep. control (no hemolysis should occur) 14 4 ce. lec. | Salt solu. control (no hemolysis should occur) NaC] solution. The determined smallest quantity established according to the primary test. Dilutions made according to Table IV. Double the quantity previously determined by titration. 5% suspension of washed sheep blood corpuscles. teore uniform and probably depends to some extent on the degree of infection present, there- fore it is advisable to use in the test such quantities of the serum as will prove sufficient for the reaction to take place. GLANDERS 139 TABLE No. VI THE FINAL TEST FOR GLANDERS. Glan- == a Tube} NaCl ee ders. | Com | Ambo-| Blood No. as heres bacteria/plement| ceptor |corpus- R ke on 1 | ram 2 extract 4 6 cles 6 emarks 1] ce. | ec. ec. ce. cee. ee. | Test tube for the dose 0.1 cc. of 1 | 0.1 1 1* 1 1 suspected serum 2 2 | OM liso 1 1 1 | Serum control for the dose 0.1 cc. suspected serum 3 1 | 0.2 1 1 1 1 | Test tube for the dose 0.2 cc. of of suspected serum 4 De) OR | asrieveeeo 1 1 1 | Serum control for the dose 0.2 ce. suspected serum 2 5 eS besaileggetne 1 1 1 1 | Control for the quantity of ex- tract used (hemolysis) Ge | sssrgwisbone aaa 2 1 1 1 | Control for the double quantity of ext. for gr. accuracy (hemo.) 7 ON Nei ck cdlletaaeetaiye 1 1 1 | Control of the hemolytic system (hemolysis) 8 Bs Maencsniacal ieyeaos Fy | lheveeersss 1 | Control of the complement (no hemolysis) 9 ON Seesect cs cea ee Says eeu 1 1 | Control of amboceptor (no hemo- lysis) 10 A | ses as ob evn canst levies Semis orc 1 | Control of salt solution (no hemolysis) 1. 0.9 per cent. NaC] solution. 2. Suspected horse serum to be inactivated for 30 minutes at 56°-57° C. in water bath, in order to destroy the complement which is present in the serum of every horse. 3. One-half of the quantity which does not prevent hemolysis and established by titration (See Table V). 4. The determined smallest quantity established according to the preliminary test (See Table III). 5. Double the quantity previously determined by titration (See Table IT). 6. 5% suspension of washed sheep blood corpuscles. The necessary quantity has been established by Schtitz and Schubert as 0.2 and 0.1 ce. placed in two different tubes. In some instances the tube containing 0.2 cc. may show a fixation of the complement, while the tube containing 0.1 cc. of the same serum may show only a partial fixation or hemolysis. In the majority of cases, however, the fixation is usually manifested in both tubes. Method of performing test. Four tubes are employed for testing the serum of each animal. Two tubes are used for the test proper and two for controls. The tubes are *Place for 1 hour in incubator or one-half hour in water bath at 37° C. 140 GLANDERS numbered 1, 2, 3,and 4. One cc. of a physiological salt solution is added to tubes Nos. land3. Twoce. is placed in tubes 2and4. The serum of the suspected horse is then added. This serum has been previously rendered inactive, that is, the complement has been “inactivated” by heating for one-half hour at 56—-57° C. One-tenth cc. of this inactivated serum is added to tubes 1 and 2. Two-tenths cc. is added to tubes 3 and 4, The antigen (glanders bacteria extract) is now added to tubes Nos. 1 and 3. One cc. of an established dilution is used. Tubes 2 and 4 are controls to see whether the suspected horse’s serum will influence hemolysis. To each tube is now added 1 cc. of a dilution of the complement (blood serum of normal guinea pigs). The proper dilution has been determined by titration as men- tioned before. A series of six controls should be made in connection with each series of tests carried out. One set of controls will do for a single day’s testing as long as the same substances (antigen, complement, amboceptor) are used in each test. It is also well to set what are called “positive” and “‘negative” controls in connection with each series of tests made. That is, the blood from a known glandered animal and likewise the blood from a healthy animal should be tested in connection with the blood from the suspected animals. The six controls mentioned above are made according to Table VI beginning with tube No. 5. Each tube is shaken carefully and placed in an incubator for 1 hour (or water bath at 37° C. for 4% hour). This is done in order to allow the union or fixing of the comple- ment which will become locked up with the antigen and the bacteriolytic amboceptor in case the suspected serum came from a glandered animal. If the bacteriolytic ambo- ceptor is not present or the suspected serum was from a healthy animal the complement will not become locked up or fixed. After the required incubation period the tubes are removed from the incubator and to each tube is added 1 ce. of the previously titrated rabbit serum (hemolytic ambocep- tor). This serum has previously been inactivated by heating to 56°-57° C. for one-half hour, provided it has not been carbolized and kept for three days. Finally 1 ce. of a 5% suspension of the washed red corpuscles of a sheep is added to each tube. The tubes are now replaced in the incubator and left for 10 hours, when the results may be read. If put in a water bath at 37° C. and left for 1 hour and then removed and kept at room temperature for from 3-5 hours the results may be read. Some workers prefer reading the results as soon as removed from the water bath. If the horse was affected with glanders, that is, the serum of the animal contained bacteriolytic amboceptor, no hemolysis will have taken place in tubes 1 and 3. The red corpuscles will have settled to the bottom and the upper liquid will be clear. The controls Nos. 2 and 4 should show complete hemolysis, that is, the fluid in the tubes should be uniformly red. If, however, the suspected serum came from a healthy horse and did not contain the bacteriolytic amboceptor, hemolysis should take place in tubes Nos. 1 and 2. Mohler gives the following advice in the interpretation of the results of the test: “Horses in which the serum produces a complete fixation of the complement in the quantities of 0.1 cc. and 0.2 cc. should be considered as glandered. “Horses in which the serum gives a complete fixation in the quantity of 0.2 cc. and an incomplete fixation in the quantity of 0.1 cc. should likewise be considered glan- dered. GLANDERS 14] “Horses in which the serum produces an incomplete fixation of the complement in the quantities of 0.1 cc. and 0.2 cc. should also be considered as glandered. “Horses in which the serum shows no fixation of the complement in either tube should be considered free of glanders.” In order to reduce the possibility of error to a minimum the agglutination test may be applied to the latter cases, and if this shows a value of 1 to 1,000 or over, the animal should be considered as glandered. However, such cases are extremely rare. Conglutination. Recently this test has been introduced by Pfeiler and Weber for the diagnosis of glanders. They claim that it has certain advantages over the complement fixation test especially in that mule serum does not have the same highly anti-complementary action as it does in the fixation of the complement. It is based on the phenomenon first described by Ehrlich and Sachs in 1902 that if one brings together the washed red corpuscles of a guinea pig, fresh horse serum, and inactive (heated to 56° C. for %4 hour) cow serum, hemoly- sis results. Bordet and Gay in 1906 showed that this action was also attended by a very vigorous agglutination of the red corpuscles and the substance in the cow serum which was responsible for the aggluti- nation was named by Bordet and Strenge ‘‘Konagglutinin.” It is purely a laboratory test and cannot be applied in the field. Glanders is to be differentiated from a variety of nasal and lymphatic disorders more or less common in the horse kind. Before the discov- ery of the specific bacterium of glanders and the specific tests, it was necessary to determine as closely as possible the differential anatom- ical characters between glanders and those of other affections, such as chronic nasal catarrh, strangles, lymphangitis, follicular ulceration of the nasal mucosa, cancer, sarcoma, actinomycosis, and the like. With the modern methods of diagnosis it is not necessary to attempt the often impossible differentiation between glanders and these lesions from the morbid changes alone. Strong has described a disease in the Philippine Islands, which first appears in nodules, that resembles glanders very closely. Itis caused by a blastomyces. It occasionally attacks cattle as well as horses. Epizoétic lymphangitis is the disease most liable to be mistaken for farcy or skin glanders. It is caused by a yeast-like fungus (Saccharomyces farciminosus). This disease was discovered by Pear- son in the State of Pennsylvania. Edwards has described a disease in mules resembling glanders. It is characterized by lymphangitis, laryngitis and extensive ulceration, gangrenous pneumonia but no formation of nodules. The mules 142 GLANDERS did not react to mallein. A bacillus resembling Bact. mallet was isolated. Tuberculosis and other lesions of the nares. Jcest has called atten- tion to the difficulty in differentiating glanders from tuberculous ulcers* on the nasal septum and tuberculous nodules in the lungs, cicatricial scars on the nasal septum due to injuries, local amyloid tumor formation and hemorrhagic nodule-like lesions on the nasal mucosa and pressure ulcers in the larynx. Parasitic nodules. In post mortem examinations, nodules are often found in the lungs, and occasionally in other organs, that are parasitic in nature but which resemble very closely those of glanders. Pathologists have long recognized parasitic nodules and their positive chemotactic action toward eosinophiles. There is a large literature on the differentiation of parasitic nodules from those of specific diseases such as glanders. Angeloff found that the “gray trans- parent nodules”’ in the lungs of horses were of parasitic nature and that the larva of a nematode usually Sclerostoma bidentatum could be found in the center of the nodules. Histologically the parasitic nodules may be recognized by the eosinophilic leucocytes which sur- round them. The studies of Moore and Fitch led to the conclusion that macroscopically it is difficult and often impossible to differentiate between the nodules due to parasites and those caused by Bact. mallet but that microscopically the lesions due to parasites are characterized by a variable eosinophilic infiltration. The existence of eosinophilia in parasitic lesions has been pointed out by Howard, Joest and others. Prevention. The physical cases of glanders are practically all spreaders and should be promptly destroyed and their stables thor- oughly disinfected including harness and watering bucket. The ex- posed animals should be tested and those that respond should be destroyed. On this point, however, there is a difference of opinion. Some authorities have affirmed that if the reacting animals are *In this country tuberculosis in horses is very rare but in Denmark it is quite com- mon. We have had one case of tuberculosis in the lung of « horse sent to the lab- oratory for diagnosis. It was sent in as a suspicious case of glanders. {A few observers have noticed eosinophilia in old nodules of supposed glanders origin. The reports, however, on this subject are not sufficiently conclusive to exclude parasites. It is not supposed that all parasites give rise to eosinophilia. Moore, Haring and Cady pointed out (Proceedings A. V. M. A., 1904) that the blood of horses infested with Sclerostoma bidentatum exhibited eosinophilia. Because of the serious- ness of infestion with this parasite, they suggested the desirability of a blood examina- tion as a procedure in eraminiug horses for soundness. GLANDERS 143 segregated, worked together in pairs, or singly and watered from individual buckets only, it is safe to keep and use them until physical symptoms develop when they should be promptly destroyed. This opinion is entertained because many horses appear eventually to recover that give a reaction to a specific test. Glanders has undoubtedly been spread by reacting animals that have been brought into a community where they developed symptoms, became spreaders and transmitted the virus to healthy horses. Among the causes for the spread of glanders are the common watering - trough*, the interchange of feeding bags and the retention of open cases of the disease. Great care should be taken to protect healthy horses against each and every channel of infection. Immunization. Marxer found that heat sterilized virus has never given any satisfactory immunizing results. Experiments with glanders bacteria treated with 80% glycerine or 10% urea, first used by Levy, Blumenthal and Marxer, have been more satisfactory. The organisms treated by these substances give a product called **Farase.”” According to Marxer it is prepared by shaking glanders bacteria in a concentration of 0.1 gm. of the bacteria to 4 cc. of a 10% urea solution for seventeen hours at 37° C. This process kills the organ- isms. “‘Farase’” has been used with considerable success in the experimental immunization of cats, guinea pigs and horses against glanders. He tried ‘“‘Farase”’ on horses that were exposed to glanders under natural conditions with promising results. Dediulin has used it with success. A large Russian breeding establishment where upwards of 3000 horses were con- stantly kept and where during the harvest season 10,000 peasant horses were hired and stabled was selected in, which to try the immunization experiment. During the year 1909, 276 horses had died of glanders. _ Six hundred horses were selected and injected with “Farase.” As a result of this work the following facts were noted: One year and four months after the immunization none of the treated horses devel- oped the disease, while during this time 14 new animals not treated with “Farase” which were kept with these died of glanders. In the meantime the treated horses were tested with mallein without obtaining any reactions. *Dr. Luckey reports great success in the control of glanders in Missouri by eliminat- ing the open watering fountains and providing hydrants where drivers can draw water for their horses in individual buckets. He states, “It is impossible ; to control glanders among horses which ere watered out of a common trough or basin.” Monthly Bulletin, Sept., 1914. Missouri State Board of Agriculture. 144 GLANDERS Specific biological treatment. A number of serums and vaccines have been tried but as yet they are unsatisfactory. A few workers have advocated the repeated injection of mallein as a remedy. The immunity established by an attack of glanders seems to be very transitory. Control. The laws of each state and country prescribe the course to be followed when glandered horses are encountered. It is a report- able disease and practitioners should conform to the law and regula- tions regarding it. With glanders, as with other infectious diseases, it is not possible to state what consideration shall be given the occult cases. The disease is governed by definite Jaws of nature and not until these are accurately interpreted can the best procedure be formulated for its control. The more general practice is to slaughter the reacting animals and thoroughly disinfect the premises. The consensus of opinion seems to be that it is not safe to keep occult cases. REFERENCES 1. Aneetorr. Die grauen durch scheinenden Knétchen in den Pferdelungen und ihre Beziehung zu der Rotzkrankheit. Arch. f. wiss u. prak. Tierch., Bd. XXXIV (1908), S. 4. 2. Bases. Observations surlamorve. Arch. de Méd. erpér. etd’ Anat. path., Vol. III (1891), p. 619. 3. Berns AND Way. Practical Application and Results of the Agglutination Method of Diagnosing Glanders in One Hundred and Fifty-two Cases. Amer. Vet. Rev., Vol. XXX (1906), p. 822. 4. Bonomer. Ueber die Schwankungen des Agglutinin und Prazipitingehaltes des Blutes wahrend der Rotzinfektion, Centralbl. f. Bakt., Bd. XX XVIII (1905), S. 601. 5. Borpet anp StRENG. Des phénoménes d’absorption et la conagglutinine sérum de beeuf. Centralbl. f. Bakt., Bd. XLIX (1909), S. 260. 6. Bourcet er MEry. Sur le séradiagnostique de la morve. La Semaine Med., 1898, p. 61. 7. Butter. Glanders. Bulletin No. 16. Miss. Agr. Exp. Station, 1891. 8. Cary. Glanders. Bulletin No. 35. Ala. Agr. Expt. Station of the Agricultural and Mechanical College, 1892. 9. Dawson. Equine glanders and its eradication. Bulletin No. 77. Florida Agric. Exp. Station, 1905. 10. DeScuwernitz anp Kitporne. The use of mallein for the diagnosis of gland- ers in horses and experiments with an albumose extracted from cultures of bacillus mallei. Am. Vet. Review, Vol. XVI (1892), p. 439. 11. Epwarps. A disease of mules simulating glanders. Veterinary Journal, Vol. LXIX (1915), p. 70. 12. Firen. Glanders in man. Cornell Veterinarian, Vol. IV (1914), p. 86. 13. Francis. Glanders, tests with mallein. Bulletin No. 30. Texas Agri. Exp. Station, 1894. 14. Fronner. Klinische Untersuchungen tiber den diagnostischen Wert der Ophthalmoreaktion beim Rotz. Monats. f. prak. Tierheilk., Bd. XXTII (1912), 8. 1. GLANDERS 145 15. Froruinauam. The diagnosis of glanders by the St thod. z Medical Research, Vol. VI (1901), p. 331. ‘ FON ee Caer 16. Gatu-Vauerto. Contribution Al’ étude de |: hologie du Bacill lei. Centralbl. fiir Bakt., Bd. XXVI (1899), S. 177. ii, Pee Oeienr etee ee Hicerns. Glanders and mallein. Proceedings Amer. Vet. Med. Asso., 1904, p. 135. 18. Huntine. Glanders. A clinical treatise. London, 1908. 19. Jost. Zur Frage der lokalen Eosinophilie bei zooparasitaren Organerkrankun- gen. Deut. Tier. Woch., Jr., 17 (1909), S. 346. 20. Jost. Uber einige rotzahnliche Erkrankungen des Respirations wege des Pferdes. Zerts. f. Infektionsk. parasit. Krankheiten u. Hygiene, Bd. XVI (1915), p. 238. 21. Lancer. Untersuchung tiber die differential diagnostische Bedeutung der Pease nese u.s. w. Monatshefte fiir prak. Tierheilkunde, Bd. XVI (1905), S. 22. LoxFFLER AND Scnurz. The bacillus of glanders. Deutsche Med. Wochen- schrift, Dec., 1882. Translated, Bd. CXV (1886), New Sydenham Society. 23. Lorenz. Versuche tiber den diagnostischen Wert der Ophthalmoreaktion beim Rotz. Berl. Tier. Woch. Jahrg., 29 (1913), S. 252. 24. M’Fapyean. The pulmonary lesions of glanders. Jour. of Compar. Path. and Therap., Vol. VIII (1895), p. 50. 25. M’Fapyean. Glanders. Jour. Compar. Path. and Therap., Vol. XVII (1904), p. 295. 26. Marxer. Die aktive Immunisierung gegen Malleus. Archiv. f. wissensch. u. prak. Tierheilk., Bd. XLI (1914-15), S. 272. 27. Mounier anp Ercayorn. The diagnosis of glanders by complement fixation. B.A. I. Bulletin No. 136, 1911. 28. Moore anp Fircu. The differentiation between nodules due to glanders and those caused by parasites. Report New York State Veterinary College at Cornell Univer- sity, 1912-13, p. 115. 29. Moors, Taytor anD Gittner. The Agglutinaticn Method for the Diagnosis of Glanders. Amer. Veter, Rev., Vol. XXX (1906), p. 803. 30. Nocarp. The value of mallein as a means of diagnosis in doubtful cases of - glanders. Jour. Compar. Path. and Therap., Vol. VIII (1895), p 227. 31. PremeR aNnp Weser. Vergleichende Untersuchungen der Sera von 100 Pferden mittels der agglutinations—, Komplement-ablenkungs-und Konglutinationsmethode zur Erkennung der Rotzkrankheit. Zeits. f. Infekkrankh. der Haust., Bd. XII (1912), S. 397. 32. Rapireaux. Serum diagnosis of glanders. Abstract Jour. Compar. Path and Therap., Vol. XVI (1903), p. 59. Orig. Jour. de Méd. Vét., 1902. 33. Reynotps. State control of glanders in Minnesota. Jour. of Compar. Med. and Vet. Archives, Vol. XX (1899). 34. Roprns. A study of chronic glanders in man with report of a case. Studies from the Royal Victoria Hospital, Montreal, Vol. 2, No. 1. 35. Ruruerrorp. Glanders. Proceedings Am. Vet. Med. Asso., 1906, p. 215. 36. Scunurer. Die Diagnose der ansteckenden Tierkrankheiten mittels der neueren Immunitatsreaktionen. Ninth International Tierdrat-Kongress im Haat., 1908. 37. Scuutz unp Scuusert. Die Ermittelung der Rotzkrankheit mit Hilfe der Komplementablenkungsmethode. Archiv. f. Wissensch. u. Prak. Tierheilkunde., Bd. XXXV (1909), S. 44. 38. Scutrz. A contribution to the subject of glanders. Jour. of Compar. Path. and Therap., Vol. XI (1898), p. 1. 39. Scnurz. Zur Lehre vom Rotze. Archiv. fiir wiss. u. prakt. Thierheilkunde, Bd. XXIV (1898), S. 1. 146 TUBERCULOSIS 40. Scuurz unp Mirssner. Zur Serodiagnose der Rotzkrankheit. Archiv. fiir wiss u. prakt. Tierhielkunde, Bd. XX XI (1905), 8. 353. 41. Smirg. On the influence of slight modifications of culture media on the growth of bacteria as illustrated by the glanders bacillus. Journal of Comparative Medicine, Vol. XI (1890), p. 158. 42, Srrone. Preliminary report of the appearance in the Philippine Islands of a disease clinically resembling glanders. 1902. No. 1, Bureau of Government Labora- tories, Manila. $ 43. Srravuss. Sur un moyen diagnostique rapide de la morve. Arch. de Méd. expér. etd’ Anat. path., Vol. III (1889), p. 460. 44. Van Es. Glanders. Bulletin No. 85, N. Dak. Agric. Exp. Station. 1909. 45. Way. The practical application of the agglutination method for the diagnosis of glanders. Am. Vet. Review, Vol. XXXI (1907), p. 709. 46. Wuerry. Glanders: Its diagnosis and Prevention. Bulletin No. 24. Bureau of Government Laboratories, Manila. 1904. 47. Wrirams. Glanders. Bul. No. 4. Mont. Agr. Exp. Sta., 1894. 48. Wricut. The histological lesions of acute glanders in man and of experimen- tal glanders in the guinea pig. Jour. of Exp., Med., Vol. I (1896), p. 577. TUBERCULOSIS Synonyms. Consumption; pearl disease; grapes; phthisis; scrofula; tabes; “The great white plague.” Characterization. Tuberculosis is an infectious disease of man and domesticated animals. Cattle and swine suffer most, but, under favorable conditions, all species including fish and amphibians are attacked. It is a disease of slow development, involving either - primarily, or in association with other organs, the lymphatic system. It is characterized in the beginning by the formation of small, non- vascular nodules, or tubercles which have a tendency to central degeneration. It destroys life by a chronic and long continued systemic poisoning and by the destruction of tissue in organs neces- sary to life. It is caused by Bacterium tuberculosis. History. Tuberculosis is one of the oldest diseases affecting cattle of which there are identifying records. It seems to have been known to the Jewish people during their Egyptian captivity and the ecclesias- tical laws for many centuries contained numerous enactments against the consumption of flesh from tuberculous animals. In 1370, it was forbidden in Munich to have on sale the flesh of animals affected with tuberculosis. A number of other cities passed similar ordinances. In 1702, Florinus described the disease and emphasized the then existing opinion that it was identical with syphilis. This led to the practice of destroying all tuberculous animals. In 1783, the Berlin Board of Health rejected the theory of the connection of tuberculosis TUBERCULOSIS 147 and syphilis and declared the flesh of affected animals to be fit for food. This led finally to the changing of all laws throughout Prussia against the use for food of flesh from animals affected with the disease. Tscheulin, in 1816, recognized in reference to the infection of meat three degrees of bovine tuberculosis, viz.: (1), in which the tubercles were to be removed; (2), in which the diseased parts were to be destroyed and the meat sold at a low price; and (3), those cases in which the lesions were so extensive that the whole carcass must be rejected. The study of the lesions themselves gave rise to a number of beliefs concerning their nature. Thus, Virchow, Schippel and others declared that the tubercles in cattle were lympho-sarcomata. Leiser- ing considered them simply as sarcomata. Spinola and Haubner maintained that human and bovine tuberculosis were identical. In 1865, Villemin showed that tuberculosis was due to a specific infection. He produced the disease in rabbits by inoculating them with tuberculous material from human subjects. He also produced the disease by feeding experimental animals and by causing them to inhale tuberculous material. Chauveau, in the same year, produced the disease in cows. These results were soon confirmed by Klebs, Cohnheim and Gerlach. These experiments, in which the disease was produced in one species with tuberculous material from another, followed by the discovery by Koch of the specific bacterium of the disease, led to the view that tuberculosis in all species of mammals was identical. This generally accepted belief caused sanitarians to look upon tuberculosis in cattle as a great menace to public health. The result was that during the closing decade of the last century this disease in cattle was treated more vigorously as a menace to the human species than as a destructive disease of animals. In 1896, Dr. Theobald Smith pointed out that for certain animals the tubercle bacteria from cattle were more virulent than those from man and further that there were certain morphological and cultural differences existing between them. In 1898, he published the results -of a more extended series of investigations. Since that time a number of investigators have arrived at the same conclusion. The fact has come to be well known that certain differences exist between the bacteria of tuberculosis found in the human and in the bovine species. Koch’s experiments reported at the tuberculosis congress in London in July, 1901, give additional evidence of a difference in virulence for experimental animals of the bacteria of human and of bovine 148 TUBERCULOSIS tuberculosis. To what extent man becomes infected from the bovine variety cannot be stated, but the accumulating evidence tends to the conclusion that bovine tuberculosis is of less significance in its influence upon public health than was formerly thought, and of more importance as a rapidly spreading and destructive disease among cattle. Concerning its transmission, the conclusion seems to be warranted, that the virus of tuberculosis spreads very largely among men and cattle from individual to individual of the same species rather than from one species to the other. Swine are often infected from cattle. Geographical distribution. Tuberculosis is an exceedingly wide- spread disease. In earlier times it was quite prevalent among cattle in Central Europe. It seems to have existed in Western Asia and Northern Africa at an early date. From these centers it has spread to nearly every cattle raising country of the world. Its rapid spread during the last fifty years is attributed to the increase in cattle exchange resulting in the introduction of tuberculous animals into healthy herds. It is stated that in many countries, and in large dis- tricts within others, tuberculosis did not exist until it was introduced within recent years by the importation of diseased animals. In countries where there has been little or no importation of cattle, * and in which the native breeds still exist unchanged, as in many parts of Russia, Austria and Spain, in the northern part of Sweden and Norway, and in parts of Africa, tuberculosis is practically unknown. This is true of the cattle on the island of Jersey, where for more than a hundred years foreign cattle have not been introduced. In the United States, the disease is very widely distributed. It is found to a considerable extent in certain localities where the climatic conditions seem to be beneficial for tuberculous people. The explana- tion for this seems to be that tuberculous animals have been intro- duced into these districts. There are, however, large areas in which it is practically unknown. The Western steers that are killed in the large slaughter houses are practically free from this disease except those that come from a few infected regions. Etiology. Tuberculosis is caused by a rod-shaped organism known as Bacterium tuberculosis. It was discovered by Robert Koch in 1882. Schiller and Toussaint had previously studied growths which seem, from the results of their inoculation experiments, to have been due to this organism. The bacterium of tuberculosis is a slender, rod- TUBERCULOSIS 149 shaped organism with rounded ends, from 2 to 5 & in length and from 0.3 to 0.54 broad. The rods are straight or slightly curved, and occur singly, in pairs or in small bundles. They do not produce spores, but vacuoles are often observed and branching forms have been described. The bacterium of tuberculosis is readily cultivated on artificial media such as blood serum, glycerinated agar and bouillon after it has been adapted to such artificial conditions.* It is, however, not easy to cultivate it directly from ordinary tuberculous lesions. Although at the time of their discovery, tubercle bacteria from man and from animals were believed to be identical, they have been found to possess slightly different characters and properties. Smith pointed out in 1898, that morphologically tubercle bacteria from cattle were shorter and thicker than those from man, that they grow slightly different on blood serum, and that they were much more virulent for cattle and rabbits than those from _ the human species. Since that time _ his conclusions have been confirmed by a number of investigators. Koch *To accomplish this necessitates a very special and careful procedure. Dr. Theobald Smith, of Harvard University (Jour. of Exp. Med., Vol. III, 1898, p. 451), has the credit of first for- mulating a method by combining details in such a manner that the procuring of culturesis, in most cases, possible. He used dog serum Fic. 25. TuBercie BAcTERIA. (a) drawn aseptically and congealed at the mini- BOVINE VARIETY FROM A YOUNG mum temperature. Other media have been CULTURE ON GLYCERIN AGAR. (b) used more recently but the fact remains that BOVINE VARIETY FROM THE MOUTH it is difficult to obtain pure cultures of tubercle OF A COW HAVING ADVANCED bacteria. It is usually necessary to inoculate PULMONARY TUBERCULOSIS. (c) guinea pigs with the original material and to AVIAN TUBERCLE BACTERIA FROM chloroform them in the early stages of the dis- A GLYCERIN AGAR CULTURE. (x ease and make cultures from the fresh, young ABouT 1000.) lesions. c 150 TUBERCULOSIS obtained like results. At present, therefore, we must look upon the tubercle bacteria coming from these different species as possessing racial or varietal differences which perhaps are the result of different life conditions. The investigations which have been made with the different forms of this organism found in tuberculosis of fowls and of fish have led a few experimenters to believe that they are simply vari- eties of the organism first described by Koch. Further inquiries are necessary to fully satisfy bacteriologists that all of these forms are thus related to the one species. There seems to be no reason for doubting that the bovine and human forms are varieties or races of the same species. The difference in the conditions of life under which they exist in the bodies of men and of cattle is quite enough to explain resulting differences in the bacteria. There seems to be a tendency for some workers on this subject to consider the varietal differences between the human and bovine to be of less significance than heretofore thought. Malm found experimen- tally that the difference between the bovine and human tubercle bacteria is not constant. He states that ‘“‘there are human tubercle bacteria that are very virulent for cattle and rabbits. There are also of the bovine type varieties that show a weak virulence for cattle, rabbits and guinea pigs.’ He believes that no distinction should be made between human and bovine varieties. Findlay and Martin found that the bovine type was more readily destroyed by daylight and drying than the human type. They explain in part by this fact the more frequent aerial infection in man. Symptoms. The symptoms vary according to the course of the disease and the location of the lesions. There isa chronic form, which is most common, and an acute form or miliary tuberculosis. The symptoms of chronic tuberculosis depend upon the location and extent of the lesions. When they are situated deeply and are not of great extent, they may not exhibit visible evidence of their presence. In such cases, the infected animal may present the picture of perfect health and show no disturbance of function. Indeed some animals, in which the lesions are both extensive and widely distributed and which have never presented noticeable signs of the disease, are slaughtered for beef without a suspicion of the presence of tuberculosis until they are examined post-mortem. Since the lesions of tuberculosis vary so much in different cases, it is not possible to give a description of what can be designated the characteristic or even the usual symptoms of this disease. In the TUBERCULOSIS 151 beginning they are largely referable to the organ affected. , There are, however, certain general manifestations that appear in most of the advanced cases, such as emaciation while the appetite continues good. This is always a suspicious indication and especially if accom- panied by cough, rough coat and tight, harsh skin. Rough or loud respiratory sounds are sus- picious, and, in advanced cases, it is often found that the ani- mal groans when pressure is brought to bear upon the chest wall. Many cases bloat habitually due to presence of enlarged glands upon the eso- phagus. Enlarged superficial lymph glands are suspicious but there are other causes for enlargement of these glands. In tuberculosis of the lungs, it may be said that coughing is the most noticeable symp- tom. It is most common after feeding, drinking, or after rapid moving following a period of repose, but some- Fie. 26. RichT LATERAL ASPECT OF PosT- times it occurs without any “prior HALF OF STHER’S HEAD. (a) LOWER ‘apparent cause. Thecoughis JAW, (b) mar passage, (c) HoRN, (d) STYLOID PROCESS OF OCCIPITAL BONE, (e) usually strong, dry and fre- parorm GLAND, (f) SUBMAXILLARY GLAND. quently of a high pitch. A. RIGHT PAROTID LYMPH GLAND. B. RIGHT é o . . POST MAXILLARY LYMPH GLAND. C. RIGHT Sometimes it is very violent, supmaxiLLARY LYMPH GLAND. THESE accompanied by protrusion of ae OFTEN THE SEAT OF TUBERCULAR ‘ ith). the tongue. Auscultation re- saa at veals modified and abnormal sounds of different kinds in the lungs; sibilant, sonorous and mucous rales are most common. A dull sound is often detected on percussion. It is also to be noted that this con- dition is of slow development and long duration, thus aiding one to distinguish it, in many cases, from bronchitis or pneumonia. Where the mediastinal lymphatic glands are enlarged and press upon the esophagus the animal bloats more or less. Chronic or habitual bloating accompanied by a good appetite and no other evi- dence of disease of the digestive tract, especially if there is shortness of 152 TUBERCULOSIS breathjand cough, may be looked upon as strongly indicative of tuber- culosis with enlarged mediastinal lymphatic glands. Enlarged tuber- cular glands along the eosphagus may also press upon that organ causing obstruction and preventing the escape of gases from the stomach. Sometimes large tuberculous masses develop on the pleura. In such cases the principal symptom is a friction sound that is heard most distinctly during inspiration. If the masses are large enough they give rise to a dull sound upon percussion. In tuberculosis of the stomach and intestines, digestion is interfered with. This gives rise to poor appetite, frequently to diarrhea and sometimes to alterna- tion of diarrhea and constipation. In tuberculosis of the peritoneum or of the lining of the abdominal cavity, the lymphatic glands of the E flank are often enlarged and hard. Sometimes this con- dition can be diagnosed posi- tively by a rectal examina- tion and the discovery of the hard, nodular masses. Tuberculosis of the liver does not give rise to symp- toms unless the disease is far advanced. In animals in which the post-pharyngeal lymphatic glands are enlarged, the breathing is harsh and noisy. In this condition there is sometimes difficulty in swal- lowing, and particles of chewed up food are occa- sionally expelled from the mouth, either voluntarily when it is found that they Fig. 27. Dorsau asPEcT OF BOVINE LUNGS. (a-a}) cannot be swallowed con- RIGHT AND LEFT CAUDAL LOBES, (b-b!) R. anp x. veniently, or by the cough- VENTRAL LOBES, (cc!) FIRST AND SECOND RIGHT ing they occasion upon CEPHALIC LOBES, (c?) LEFT CEPHALIC LOBE, (e) . TRACHEA, (X-X) REGION MOST FREQUENTLY In- Teaching the pharynx. VOLVED IN THE EARLIEST STAGES OF PULMONARY TUBERCULOSIS. THE LESIONS AT THIS STAGE ARE These enlarged glands ae USUALLY EMBEDDED IN THE LUNG TISSUE (Smith). sometimes be detected by TUBERCULOSIS 153 palpation accomplished by placing one hand on each side of the throat above the larynx and then pressing from opposite sides. Tuberculosis of the udder is detec- ted by an enlargement and hard- ening of the affected part, usu- ally by the absence of pain and the fact that the secretion is not altered until the part has been dis- eased for some time. In advanced cases, instead of milk, the udder secretes a yellowish, cloudy and sometimes flocculent liquid. In acute, rapidly developing cases, there may be pain and edema of the skin. tuberculosis the supramammary lymphatic glands, situated above the udder in the middle of the escutcheon, are enlarged and hard. If there is doubt as to the character of the disease of the udder, the milk, or possibly a piece of excised udder tissue, may be examined bacteriol- ogically. In tuberculosis of the brain, the animal is unsteady and uncertain in its movements. It lies down much of the time, is usually subject to occasional cramps and is apt to carry the head in an unusual position. Such cases are inclined to advance rapidly and terminate in death following coma or convulsions. In tuberculous disease of the bones and joints, the parts are enlarged, In nearly all cases of udder ra fy I fe Fia. TUBES OF BOVINE LUNGS SHOWING 28. TRACHEA AND BRONCHIAL ATTACHED BRONCHIAL GLANDS. (aa!) SUPPLY RIGHT AND LEFT CAUDAL LOBES, (b-b!) SUPPLY R. AND L. VENTRAL LOBES. (cc!) BRANCHES OF THE RIGHT SUPER- NUMERARY BRONCHUS, (c2) SUPPLY LEFT CEPHALIC LOBE, (d) BRANCH TO AZYGOUS LOBE, (e) TRACHEA. A. LEFT BRONCHIAL LYMPH GLAND. B. RIGHT BRONCHIAL LYMPH GLAND. C. LYMPH GLAND BASE OF SUPERNUMERARY BRONCHUS. D. GLAND OFTEN BETWEEN BRONCHI. THE GLANDS ATO D ARE OFTEN INVOLVED (Smith). there is loss of motion, pain and usually abscess formation followed by the discharge of thick yellow pus. In tuberculosis of the uterus or ovaries and sometimes in peritoneal tuberculosis of the cow, the subject is almost continually in heat. In tuberculosis of the uterus 154 TUBERCULOSIS there is sometimes a discharge of thick, yellowish material mixed with mucus. In tuberculosis of the testicles the organs become en- larged and hard. In all advanced cases, the nutrition of the animal is interfered with and, sooner or later, the “tuberculous cachexia” appears. It is, however, in many cases re- markable to note the extent of lesions in animals that are well nourished and present no external signs of disease. Animals killed in prime con- dition by the butcher are sometimes found to contain extensive and widely dis- tributed lesions of tubercu- losis. In generalized tuber- culosis, many of the symp- toms described above may occur simultaneously. The symptoms of acute miliary tuberculosis are rapid loss of flesh, depression, poor appe- tite, cough, weakness, rapid breathing, harsh respiratory Fic. 29. DORSAL ASPECT OF BOVINE LUNGS SHOW- ING POSITION OF THE POSTERIOR MEDIASTINAL GLANDS; (a, b, e, c’) CAUDAL, VENTRAL, CEPHALIC LOBEs, (f) ESOPHAGUS, (g) MUSCULAR PILLARS OF DIAPHRAGM, (h) POSTERIOR AORTA, (i) CAUDAL MARGIN OF THE LIGAMENT OF THE LUNG. A. LEFT BRONCHIAL GLAND. MEDIASTINAL GLANDS ARE SHOWN, MOST OF THEM RESTING ON THE ESOPHA- GUS. THE LARGE CAUDAL GLAND RESTING ON THE PILLARS OF THE DIAPHRAGM IS MOST FREQUENTLY DISEASED AND OFTEN ATTAINS AN ENORMOUS SIZE. THE REMAINING MEDIASTINAL GLANDS ARE ARRANGED IN TWO SETS ON THE RIGHT AND LEFT MARGINS OF THE ESOPHAGUS (Smith). charged into the blood or lymph currents. sounds, some elevation in temperature, increased pulse rate and, sometimes, enlarged lymphatic glands. The course of this form of tuberculosis is always rapid and. terminates in death. Acute miliary tuberculosis occurs when large numbers of tubercle bacteria are dis- They are then carried to other parts of the body, filtered out in the capillaries of the lungs, liver, spleen, kidneys and elsewhere, causing tubercular lesions in each of these localities. The lesion from which the infectious material entered the circulation may have been a comparatively small nodule. This form of the disease is more likely to appear in TUBERCULOSIS 155 young animals than in adults, and is more common among swine than in cattle. Morbid anatomy. The usual direct anatomical change following the invasion of tubercle bacteria is the formation of nodules or tuber- cles. A tubercle has been defined as ‘‘a small nonvascular nodule composed of cells varying in form and size with some basement sub- stance between them and with an inherent tendency to undergo central necrosis." In a large number of cases the individual tubercles are distinct and easily recognizable, while in others they are coalesced, forming a mass of necrotic tissue. The lesions vary, therefore, from Oe 990 %58 9g 0) FP LOe LLY 0 © 19.0 29 a * KS © oes gee egy ee ao 8 @ / es inom. RHI ne a eee Fig. 30. A DRAWING OF A SECTION OF VERY YOUNG TUBERCLES IN SPLEEN (Thoma). well isolated minute or larger nodules to masses or cavities containing a purulent, caseous, or calcified substance. The location of the primary lesion depends upon the channel of infection. If the specific organisms are lodged in the oral cavity or pharynx they may pass through the mucosa and be taken to some of the lymphatic glands about the head; if they are taken directly through the respiratory passages into the lungs they either develop nodules in the lung tissue proper, or they are carried through the lymphatic system to the lymph glands draining the lungs where the lesions first appear. If the specific bacteria are first lodged in the intestinal mucosa, primary tuberculous ulcers may develop or they may pass 156 TUBERCULOSIS Fig. 31. A PHOTOGRAPH OF A SECTION OF THE ANTERIOR LOBE OF A COW'S LUNG ADVANCED IN TUBERCULOSIS. THE ENTIRE LUNG TISSUE IS INVOLVED. THE TUBER- CULOUS MASSES ARE SURROUNDED IN SOME INSTANCES BY a QUITE THICK BAND OF CONNECTIVE TISSUE. A LARGE PART OF THE TISSUE IS CALCIFIED. THIS ISSHOWN BY THE LIGHT OR WHITISH POINTS. (NATURAL sIzE.) into the mesenteric lymphatics or the portal vein. It may happen that the bacteria may be carried by means of the lymph or blood TUBERCULOSIS 157 stream and lodged in any part of the body, such as the brain, kidneys, spleen, testes, ovaries, bones, joints, and subcutaneous and inter- muscular glands and serous membranes. The evidence at hand, how- ever, seems to show that in a large majority of cases the primary lesions are located in one of the following organs: (1) in the lungs or the lymphatic glands draining them; (2) in the lymphatic glands about the head; (3) in the mesenteric glands and intestines; (4) in the portal glands or liver substance itself; and (5) in the genera- tive organs and udder. It not infrequently happens that the apparent primary lesions occur on the pleura, peritoneum, meninges or synovial membranes while the organs remain freéYrom disease. In such cases the lesions consist of many tubercles varying from one to ten or more millimeters in diameter or of bunches of closely set tubercles which are more or less flattened or irregular in shape, owing to their mutual pressure. Sometimes these tubercles are attached to the serous membrane by a small, tough, fibrous pedicle: frequently, however, this is absent and the nodules rest bodily upon the membrane. The structure of the tubercle consists in the beginning of a few cells surrounding the invading specific organisms. These are soon encased by a zone of epithelioid cells and giant cells which is soon surrounded by an outer layer of round or lymphoid cells. The central portion becomes necrosed and as the nodule enlarges the central necrotic por- tion becomes correspondingly large. The histological structure of the tubercle is typically illustrated in the beginning avian tubercle. In cattle there is a strong tendency for the necrotic tissue to become infiltrated with- lime salts. In certain species a deposit of fibrous tissue in the. outer zone of the tubercle has been observed. In the smaller and more susceptible experimental animals such as the guinea pig and rabbit and frequently in swine, the lesions are of a more diffuse nature infiltrating the inter- stitial tissue with the tuberculous mass and gradually encroaching upon the parenchyma. Circumscribed tubercles may also be present. In secondary or generalized tuberculosis one or more of the organs, such as the omentum, serous membranes, or lymphatic system, may become more or less thickly sprinkled with minute grayish nodules about the size of a millet seed. These tubercles are at first almost the color of mother-of-pearl but later as the central caseous degeneration begins they become grayish. Giant cells are usually numerous. 158 TUBERCULOSIS Fic. 32. A PHOTOGRAPH OF THE TUBERCULOUS PROMINENCES OR NODULES THAT HAVE DEVELOPED ON THE PLEURA COVERING THE RIBS. (NATURAL SIZE). In studying the lesions in a fatal case of tuberculosis one may find with varying modifications one or more of the following conditions: The primary lesion may be found in any one of the organs or mem- branes. Its comparative age is determined by the character of the TUBERCULOSIS 159 anatomical changes. It may be entirely encysted, caseous or cal- careous and dead. In addition to the primary focus, there may be a succession of tubercles of various ages distributed in one or more organs. The lesions may be restricted to one organ, as the liver, in which the primary focus has spread by continuity due to its infiltrat- ing nature until the destruction of the tissues of the organ has be- come so extensive that death results. Such cases do not seem to be common. sec PT Fic. 33. TUBERCLE DISCHARGING INTO BRONCHUS. THIS SHOWS A SECTION THROUGH a BRONCHUS WHERE AT POINT (a) THE TUBERCULOUS TISSUE HAS EXTENDED INTO THE BRONCHUS MAKING IT POSSIBLE FOR THE TUBERCLE BACTERIA FROM THE TUBERCULOUS AREA TO PASS INTO THE BRONCHUS AND THROUGH IT TO THE MOUTH. FROM THE MOUTH THEY ARE DISSEMINATED WITH THE DROOLINGS OR THEY ARE SWALLOWED AND APPEAR IN THE INTESTINAL CONTENTS. (NATURAL SIZE). The primary lesion may be well marked and accompanied by miliary tubercles sprinkled extensively throughout the organs and tissues of the entire body. The lesions throughout the body may resemble each other very closely, so that difficulty may be experienced in determining the prim- ary focus. In the lungs, two distinct forms of lesions are observed. (1) The air cells may be infiltrated with the tuberculous mass spreading 160 TUBERCULOSIS directly from the primary focus. This may be purulent, caseous or calcareous. The color may be whitish, gray or of a yellowish tinge. (2) The lesions may consist of miliary tubercles. In later stages these nodules, more or less translucent, may become yellowish, caseated and calcareous in their centers. Large tubercular nodules are frequently formed by the massing of several of these minute tubercles. Fic. 34. TuBerctLosis oF THE MUCOUS MEMBRANE OF THE TRACHEA, COW. When the lungs are primarily attacked the caudal (principal) lobes are most frequently involved. Smith considers the seeming predilec- tion for the larger lobes to be due to mechanical conditions. The writer has found, however, that in certain herds which have been killed after the tuberculin test. the primary and only lung lesions were in the TUBERCULOSIS 161 ventral and cephalic lobes. It is important to note that usually the bronchial glands are also involved. When the pleure are affected the lesions consist of nodules varying in size from that of a millet seed to a large pea, sprinkled more or less thickly on one or both of the visceral or parietal surfaces. These form the “pearl disease,” Perl- sucht, of the German and the “grape disease” of the English writers. If they become confluent, large masses are found. Jcest and Mar- janen found that in serous tuberculosis in cattle, there are produced non-specific inflammatory new formations which become infected with tubercle bacteria and result in the formation of the “pearl” nodules. Tuberculosis of the thoracic glands is very common and usually accompanies lesions in the lungs; but the lungs may be healthy and the glands involved. (See figures for location of glands.) The primary lesions may be and often are found in the lymphatic glands about the head. In rare cases the lesions are found in the mucous membrane of the trachea. In these cases the mucosa is often wrinkled. In some cases very small lesions are found discharging into a bronchus. In the abdominal cavity the organs most frequently involved are the peritoneum, mesenteric lymph glands, portal lymph glands and liver. The kidneys, spleen, ovaries and uterus are more rarely the seat of tuberculous lesions. Ulcers in the intestine have not been common in the writer’s observation. The ulcers in the cases observed have been isolated with elevated borders and a depressed center. Sections show that the tuberculous infiltration extends outward and to a certain extent undermines the mucosa. Tuberculosis of the testes is sometimes found. The udder becomes the seat of tuberculous deposits in a small percentage of cases. It is more often affected in cases of generalized tuberculosis. M’Fadyean finds udder tuberculo- sis in from 1 to 2 % of the cows; Poels 0.9%; Vallée and Villejean 5.3 to 6.5%; and Bergman 3.5%. When the primary infection is restricted to a single focus the disease is said to be localized. When the specific bacteria are spread from the primary lesions through the agency of the lymph and blood streams, sprinkling other organs with the infecting bacteria, each of which becomes the starting point for the development of a new tubercle, the disease has become generalized.* *The Federal meat inspection regulations state that animals affected with “extensive or generalized tuberculosis’ are to be condemned. 162 TUBERCULOSIS Fic. 35. A PHOTOGRAPH OF A PART OF THE OMENTUM SHOWING A LARGE NUMBER OF SMALL MORE OR LESS FLATTENED TUBERCLES SCATTERED OVER THE SURFACE. (SLIGHTLY REDUCED IN SIZE). It was formerly considered that when the Jesions existed in both of the large (abdominal and thoracic) cavities of the body the disease was generalized. It is possible, however, for it to be generalized when the lesions are restricted to the organs of one cavity, as the TUBERCULOSIS - 163 secondary seeding with the bacteria that have escaped from a primary focus through the circulation may be restricted to the cavity in which the first lesions developed. It seems better, therefore, to accept Ostertag’s views and classify local and generalized tuberculosis in ac- cordance with the nature of the lesions rather than their distribution in the body. The fact is worthy of consider- ation, that very often cattle killed after reacting to tuberculin do not show extensive distribution of lesions. Frequently animals are killed soon after infection has taken place, in which case the lesions are restricted to a single lymphatic gland or other organs. In other cases old lesions of considerable proportion are found. Diagnosis. Tuberculosis is diag- nosed by the symptoms, lesions, by finding the tubercle bacterium, and by the use of tuberculin. The sera tests thus far have not been satis- factory. Lesions. The lesions in tubercu- losis are usually sufficiently charac- teristic to enable a diagnosis to be made from the gross examination of the affected organs. There are cases, however, where they are atypical and a diagnosis cannot be made from their appearance. Etiology. In many cases the spe- cific bacterium may be found by Fic. 36. A PHOTOGRAPH OF A SHORT STRIP OF THE SMALL INTESTINE OF A COW SHOWING SEVERAL SMALL AND ONE LARGE TUBERCULOUS ULCERS (NATURAL SIZE). staining films made from the lesions with the ordinary tubercle stain.* In young lesions this is usually not difficult but in old ones it is often *4 method for staining tubercle bacteria. Stain the preparation with fresh carbol 164 * TUBERCULOSIS impossible. Sections of the tissues containing young lesions properly stained are often helpful. In case the tubercle bacteria are not found in smears or sections, guinea pigs should be imoculated.* If the inoculated guinea pigs are chloroformed in about 25 days young tubercles can usually be detected in the adjacent lymph glands from which cultures may be made. The tubercle bacteria are usually readily demonstrated by making film preparations from the lesions. fuchsin. Place a few drops of the stain on the film side of the cover-glass preparation and hold it over a flame with forceps until steam is given off. Allow the hot stain to act for from 3 to 5 minutes, or the preparation may be floated on the carbol fuchsin in a watch glass without heat. In this case it is allowed to act for from 10 to 15 minutes. The preparation is then rinsed in water and decolorized by treating it with a 10% solution of nitric or sulphuric acid for from 14 to 1 minute. It is again rinsed in water, when it is ready for examination. It can be dried and mounted permanently in bal- sam. The tubercle bacteria should be stained a deep reddish color. All other bacteria or animal] tissue in the preparation should be nearly or quite decolorized. If desired, a counter-stain, such as alkaline methylene blue, may be used after decolorizing; that is, the preparation should again be stained for about 1 minute in alkaline methylene blue, rinsed in water, and examined as before. In these preparations the tubercle bacteria are red and the other organisms and cells are blue. A counter-stain is of little value in preparations made for simple diagnostic purposes. When a counter-stain is desired Gabbett’s decolorizing and counter-staining solution is very convenient. GABBETT’S SOLUTION Methylene blue (powder) ............00 000 ccc ccc eee eee eee 2 grams TOO Sua PUTO ACL: oy So 2 ccc ce a seacesecniden 4 epergud Sob eotlpctoc ses

Fig. 40. TEMPERATURE CURVE OF AHOG. DOTTED LINE A REPRESENTS TEMPERATURE OF A HOG FOR 24 HOURS BEFORE THE INJECTION OF TUBERCULIN. THE FULL LINE A REPRESENTS THE TEMPERATURE OF THE HOG FOR 24 HOURS AFTER THE INJECTION OF TUBERCULIN (Schroeder). absence of generalization. Zschokke has called special attention to the localization of tuberculous lesions in the head of swine, especially in the nares and brain. Diagnosis. Tuberculosis is diagnosed in swine by the same methods as in cattle although several of the special tests are not applicable to swine. The most usual means are (a) by the lesions; (b) finding the specific bacterium; and (c) the use of tuberculin either subcutaneously or by the intradermal test. TUBERCULOSIS IN OTHER MAMMALS 181 TUBERCULOSIS IN OTHER MAMMALS Genera affected. It is stated that all species are sometimes attacked. Tuberculosis in the horse is rare, although a total of many cases have been reported. Bang has collected twenty-nine cases. In Saxony .08 per cent. of the horses (3,500) that were slaughtered were tuberculous. In this and most countries there are no reliable Fic. 41. LUNG OF HORSE CONTAINING TUBERCLES (a). statistics respecting the extent of the disease in this species. M’Fad- yean has pointed out the fact that in a considerable number of cases of equine tuberculosis, where the horses have been fed milk from tuber- culous cows, the morbid anatomy differs but slightly from that in tuberculous cattle. Recently several authors have reported isolated cases in Europe. In this country horses are practically free from it. We have seen but one case. Sheep and other domestic animals are reported to suffer more or less extensively from this disease. All of the so-called tuberculosis in 182 AVIAN TUBERCULOSIS sheep that I have examined proved not to be tuberculosis but the “nodular disease’’ caused by an animal parasite (Oesophagostoma columbianum). A few cases, however, have been reported. Tuberculosis in dogs and cats is quite rare but several cases in each genus are on record. Schlesinger reports a case of miliary tuberculo- sis in a dog with ulcerative endocarditis. Blair has reviewed the litera- ture on this subject and given the results of his investigations. Fic. 42. SPLEEN HORSE SHOWING TUBERCLES NATURAL SIZE AVIAN TUBERCULOSIS History. In America, tuberculosis in fowls was described in 1900 by Pernot in Oregon and Burnett in northern New York. In 1903 Moore and Ward found the disease in California, where in certain flocks it was very destructive. It was recognized by the owners as “spotted liver,” going light, and rheumatism. In Europe it has been known for many years. There is an extensive literature on this subject. Symptoms. The symptoms that are quite constant are emacia- tion, which in advanced cases becomes extreme, and anemia. The comb, skin, and visible mucosa about the head are usually pale. As the course of the disease advances the feathers become ruffled and the AVIAN TUBERCULOSIS 183 fowls are weak, dumpish and move about very little. The eyes are bright in most cases until the end is near. The appetite is good, and the fowls eat ravenously until a few days before death. The tempera- ture is in most cases within the normal limits, rarely it is subnormal. The blood is pale. The hemoglobin varies from thirty-five to seventy per cent. as tested with Gowers’ hemoglobinometer. The red blood corpuscles vary from 1,010,000 to 2,600,000 per cubic millimeter. There appears to be a slight increase in the number of white corpuscles especially of the eosinophiles. — Tuberculous fowls are often lame. Pernot mentions this as one of the important symp- toms in the cases he observed. It is due to joint lesions in some cases. In others it ap- pears to be due to extensive lesions in the viscera. | | The avian variety of tubercle bacteria resembles quite closely those of the human and bovine varieties in size and general | morphology as they are found | in the tissues of the fowl. A : measurement of over two hun- dred individual organisms in cover glass preparations made ‘ ie i -/.| directly from organs of fowls A PHOTOGRAPH OF A TUBERCU- gave the following: In the liver Pe eee the length varied from 1.2 to 3.5¥.1n the spleen and in the skin they varied from 1 to 4 in length. A general average gave a length of 2.74. They often appear in these preparations in dense masses. Chains made up of a number of short elements are rarely present. Granules are occasionally observed. In the preparations from the skin a considerable num- ber of them contain polar granules and not infrequently ‘three such bodies were noticed in a single individual. Perhaps the most striking feature concerning these organisms in the tissues is their enormous numbers. Sibley has called attention to the similarity of avian tubercle bacteria to those of leprosy in that they multiply to 184 AVIAN TUBERCULOSIS such enormous numbers without a pronounced breaking down of the tissues. This variety can be obtained in pure cultures in about 20 per cent. of serum tubes inoculated directly from tuberculous lesions in fowls (Moore). It grows readily in glycerin agar, Dorset’s egg medium, in glycerin bouillon and on potato. Fowls inoculated in the abdominal cavity or subcutaneously with from one-half to one cubic centimeter of a glycerin bouillon culture develop either localized or generalized tuberculosis in from six weeks to three months, but a longer time is ordinarily necessary to kill them. Rabbits and guinea pigs are not readily infected by the inoculation of pure culture. Moore and Ward failed to produce any tuberculous lesions in these species by this method. Morbid anatomy. The lesions are widely distributed, and vary much in their location in different individuals. The liver is most frequently involved. The spleen, intestines, mesentery, kidneys, lungs and skin are affected in order mentioned. The appended table gives the distribution of the lesions in 17 cases observed by Moore. THE DISTRIBUTION OF LESIONS IN TUBERCULOUS FOWLS a ORGANS INVOLVED.* ° & 2| 8 ate Bet 5 By 3G | Liver | Spleen Tates- |Mesen- lcidney | Ovary | Lungs | Bones | Skin Tl Ke evesex DON tihtaialle armnaighaliiialsine lltsndeullieua slattlaseuund Weenies lanes ON DP |i ves cl ee hpeale econ allen a's allen eal a edie oad es |Sewea ai een es XxX S) [1065 | MCX. cl ea saleensss awe clase adelssaseelecrs eel seaaws 4, K_ |107 XX| XX elke cccth va] k Scensesl| eaasenenccg eevee ate averacdna [oseuoed 5) K /|105.2 | XXX)...... DO A calll eadssccumailteraataaselicis svenaa lcs atler [MusAyalals 6) KK OUA: V OER DOG os eral tres [eeteaire |e satin alls anata [fe aecencie [he wanecaie TK, [LOTD: |e seal ossas XXX} XXX]}...... DO osmefemaeetdlyinaaes 8 Dr lapsere XxX x x x DE sclashen'al le ainarel| & aeantetell te a overess OF Dt |e ccsneleamealeveecelaseamaley oye SE comme sl serene bere 10} K {108.4 | XXX PG Career [cormaceetre ear concanteal Parrareeramn) bse creer neers (Rares li; K |106.4 | XXX x b,¢ Paeaet en ered Pert etree peered ban ae 12) K |107.4 | XXX) XX >. eee XXX]...... MEK ceca y| seicacecs 13} K_ {107 OER Sisdase sullen dytatell ace tecanel a aan bette cae deametete’S leereeroars leneteate 14) KO L076 XN Soon aamanalll anaes leave gaccltegews «lheetaind [amie ae aaa 16) K |1068°) XXX) XX eccealecgezslecvecalervers leomees loeaavelekoeas 16), “D! |eiscad bzeuis esas [paeme | is acti lien ee ectcat Neteeae eed: seca | svtsannan a 171 K_ /105 XXX| XXl...... 2,9, 4 eran ener eee rea eae ee *The relative numbers of tubercles are indicated by the number of Xs. XXX indicates an extensive invasion, XX a less number of tubercles, and X very few, Figures 43 and 44 show extent of lesions represented by XXX. The tubercles in the earlier stages of the disease, especially in the liver, are small greyish points varying from 0.25 to 1.0 millimeter in AVIAN TUBERCULOSIS 185 diameter. In advanced cases they are larger. They have a cheesy consistency, and are easily removed from the surroun ding tissue. The removed, necrotic nodules have a roughened surface. The color is greyish or whitish in the early stages, but in the later ones it changes to a yellowish tint. Occasionally there are two distinct crops of tubercles, one consisting of nodules 4 to 6 millimeters in diameter and separated by a centimeter or more, and the other of closely set grayish Fig. 44. A PHOTOGRAPH OF A SECTION OF TUBERCLE FROM A FOWL, SHOWING THE NECROTIC CENTER AND SURROUNDING ZONES. ENLARGED. tubercles 0.25 to 0.5 mm. in diameter. In some cases the tubercles are few in number but larger in size. The liver cells between the tubercles are usually in a state of more or less degeneration, and frequently fat globules are numerous. The blood spaces are more than normally distended with blood. The lesions in the spleen, like those in the liver, consist of minute or larger tubercles of a grayish or of a yellowish tint. The central portions of the larger tubercles are often homogeneous, darker in color and more or less hyaline in appear- ance and consistency. The tubercular growths in the intestine start in the walls of the intestine. They present a glistening appearance, grayish in color 186 AVIAN TUBERCULOSIS and firm to the touch. Frequently they are confluent. When single they vary from 1 to 10 mm. in diameter. They are usually sessile on the intestine but on the mesentery they are frequently peduncula- ted, varying from 2 to 5mm. in length. On section the young tubercles exhibit a grayish, glistening surface, but the more advanced nodules contain recognizable necrotic centers. In the larger tuber- cles on the intestines the necrotic centers frequently open into the lumen. Fig. 45. A PHOTOGRAPH OF A TUBERCULOUS MESENTERY OF A FOWL. THERE ARE A FEW SMALL TUBERCLES ON THE INTESTINE. TUBERCULOSIS 187 The skin lesions consist of a cellular infiltration usually abont the root of the feathers. Frequently the nodules become confluent. They may or may not involve the subcutaneous connective tissue. The microscopic examination of the tubercles of the liver shows them to consist of a necrotic center surrounded by an irregular zone of epithelioid and giant cells. This is surrounded by a band of tissue consisting for the greater part of liver cells, more or less disintegrated free nuclei and a few infiltrated round cells. This zone is circum- scribed by a narrow reactionary band consisting very largely of round cells. The structure is constant in both small and large tubercles, and not strikingly different from the structure of tubercles in certain of the mammals. The larger nodules seem in some instances to be the result of a continuous growth of a single tubercle, and in others to have resulted from the coalescence of a number of smal! ones. The necrotic center and reactionary zone of round cells are beautifully demonstrated by their reaction to nuclear stains. REFERENCES 1. Apams. On the significance of bovine tuberculosis and its eradication and prevention in Canada. Canadian Jour. of Medicine and Surgery, Dec., 1899. 2. Assmann. Vergleichende Untersuchungen tber die thermische Tuberkulinprobe und die Phymatin-Opthalmoreaktion. Berliner Tierdratlich. Wochensch., Bd. XXVII (1911), §. 449. 3. Buarr. Tuberculosis in the dog and cat. Proceedings of the N. Y. State Vet. Med. Soc., 1915. 4. Curtice. The detection of tuberculosis in cattle. Annual Report, Bureau of Animal Industry, U. 8. Dept. Agric., 1895-96. 5. Dorset. Experiments concerning tuberculosis. Bulletin 52. Bureau of Animal Industry, 1904. 6. Exner. Suggestions for a uniform system of interpreting the tuberculin reaction in cattle. The Jour. Compr. Path. and Therap., Vol. XVIII (1905), p. 224. 7. Frypuay anp Martin. The effect of daylight and drying on the human and bovine types of tubercle bacilli. British Med. Jour., 1915, p. 110. 8. Fora. Tuberkulipnroben nach Moussu und Mantoux. Berliner tierdrztlich. Weocehensch., Bd. XXV, (1909), S. 727. 9. Harrnc. Bovine tuberculosis investigations at the University of California Farm. Proceedings of the A.V. M. A., 1910, p. 306. : 10. Jorst AnD Marganen. Histologische Studien tiber die Serosentuberkulose des Rindes. Zeitsch. fiir infek. Krankheiten, Bd. XV (1914), 5. 1. 11. Kocu. The etiology of tuberculosis. Mitt. aus. dem. Kaiserl. Gesundheitsamte, Bd. II (1884). Translated in Vol. CXV, (1886). New Sydenham Society. 12. Kocu. The combating of tuberculosis in the light of the experience that has been gained in the successful combating of other infectious diseases. Amer. Vet. Rev., Vol. XXV (1901), p. 441. 13. Luckey. The intradermal tuberculin test. Amer. Vet. Rev., Vol. XLI (1912), p. 316. 14. M’Fapyean, Saeatuer, Epwarps anp Minerr. Experiments regarding the vaccination of cattle against tuberculosis by the intravenous injection of tubercle bacilli of the human and avian types. Journ. of Comp. Path. and Therap., Vol. XXVI (1913), p. 387. 188 TUBERCULOSIS 15. Maim. Ueber die Typen und Uebergangsformen des Tuberkelbacillus. Deut. tier. Wochensch., Bd. XXI, 8. 746. 16. Matm. Ueber die sogenamten bovinen u. humanen Typen des Tuberkelbac- illus. Contralb. f. Bakt., Bd. LXV. 17. Meyer. The conjunctival reaction for glanders. Jour. Infect. Dis., Vol. XII (1913), p. 172. 18. Mouter. Infectiveness of milk of cows which have reacted to the tuberculin test. Bulletin 44. Bureau of Animal Industry, 1903. 19. Moutmr. Tuberculosis in hogs, with special reference to its suppression. Amer. Vet. Rev., Vol. XXXII (1907), p. 176. 20. Monier anp Wasnpurn. A comparative study of tubercle bacilli from varied sources. Bulletin 96. Bureau of Animal Industry, 1907. 21. Moore anp Dawson. Tuberculosis ‘in swine, the nature of the disease with a report of three cases. Annual Report, Bureau of Animal Industry, U. 8. Dept. Agric., 1895-96. * 22. Moore. The preparation of tuberculin, its value as a diagnostic agent, and remarks on the human and bovine tubercle bacilli. Trans. of the Med. Society of the State of N Y., 1900. 23. Moors. Areport on bovine tuberculosis. New York State Dept. of Agric., 1903. 24. Movussu anp Manroux. On the intradermal reaction to tuberculin in ani- mals. Comp. Rend. Acad. Sct., Paris, Vol. XVII (1908), p. 502. Abs. in Expt. Sta. Record, Vol. XXI (1909), p. 582. 25. Movussu anp Mantoux. Sur lintra-dermo-reaction 4 la tuberculine chez les animaux. Proceedings 6th International Congress on Tuberculosis, Vol. IV, p. 821. 26. Nocarp. The animal tuberculosis. New York. 27. Pearson. The Pennsylvania plan for controlling tuberculosis of cattle. Proc. Amer. Vet. Med. Asso., 1899. 28. Prarson. Tuberculosis in cattle and the Penn. plan of its repression. Bulletin 75. Penn. Dept. of Agric., 1901. 29. Prarson. The repression of tuberculosis in cattle by sanitation. Bulletin 74. Penn. Dept. of Agric., 1901. 30. Prarson. The artificial immunization of cattle against tuberculosis. Amer. Vet. Rev., Vol. XXTX (1905), p. 543. 31. Ravensy. The dissemination of tubercle bacilli by cows in coughing a possible source of contagion. Univ. of Penn. Med. Magazine, Nov., 1900. 32. RaveneL. The comparative virulence of the tubercle bacillus from human and bovine sources. Univ. of Penn. Med. Bulletin, Sept., 1901. 33. Ravenet. The intercommunicability of human and bovine tuberculosis. The Uni. of Penn. Med. Bulletin, May, 1902. 34. Repp. Transmission of tuberculosis through meat and milk. American Medicine, Oct. 6, Nov. 2, 1901. 35. Satmon. Legislation with reference to bovine tuberculosis. Bulletin 28. Bureau of Animal Industry, U. S. Dept. of Agric., 1901. 36. Satmon. The tuberculin test of imported cattle. Bulletin 32. Bureau of Animal Industry, U. S. Dept. of Agric., 1901. 37. Satmon. Bovine and human tuberculosis. Proceedings Amer. Vet. Med. Asso., 1903, p. 436. 38. Satmon. Tuberculosis of the food-producing animals. Bulletin 38. Bureau of Animal Industry, 1906. 39. ScHROEDER AND Corton. The relation of tuberculous lesions to the mode of infection. Bulletin 93. Bureau of Animal Industry, 1906. 40. ScHROEDER anp Corron. Experiments with milk artificially infected with tubercle bacilli. Bulletin 86. Bureau of Animal Industry, 1906. 41. ScHRoEDER AND Mouter. The tuberculin test of hogs. Bulletin 88. Bureau of Animal Industry, 1906. TUBERCULOSIS 189 42. Smirg. Investigations concerning bovine tuberculosis with special reference to diagnosis and prevention. (Pathological part.) Bulletin No. 7, Bureau of Animal Industry, U. S. Dept. of Agric., 1894. 43. Smira. A comparative study of bovine tubercle bacilli and of human bacilli from sputum. The Jour. of Exper. Med., Vol. III (1898). 44, Smiru. The thermal death point of tubercle bacilli in milk and some other fluids. The Jour. of Exper. Med., Vol. IV (1899), p. 217. 45. Smira. The channels of infection in tuberculosis, together with some remarks on the outlook concerning a specific therapy. Trans. Mass. Med. Soc., 1907. 46. Upatuanp Brrcu. The diagnosis of open cases of tuberculosis. Report N.Y. State Vet. College at Cornell Univ., 1913-1914, p. 55. _ 47. Warp anp Baker. Experiments with the intradermal test for tuberculosis in cattle. Proceedings Am. Vet. Med. Asso., 1910. 48. Witts anp Lincu. Delayed reactions following injection of tuberculin. Report of U. 8. Live Stock Sanitary Association, 1913. 49. Writs anp Lincu. Delayed reaction’ following injection of tuberculin. Cornell Veterinarian, Vol. IV (1914), p. 16. REFERENCES AVIAN TUBERCULOSIS 1. Bray. Tuberculosis in chickens. Jour. Compar. Med. and Vet. Archives, Vol. XVII (1896), p. 461. 2. Burnett. Tuberculosis in chickens positively identified in New York. Am. Vet. Review, Vol. XXX (1907), p 312. 8. Captor. Sur la tuberculose du cygne. Bul. de la Soc. Cen. et Méd. Vét., Vol. XLIX (1895), p. 570. 4. Captot, GinBerT anpD Rocer. Inoculation of the tuberculosis of gallinaceous to mammalia. Amer. Vet. Review, Vol. XX (1896-7), p. 225. 5. Capiot, GILBERT ET Rocer. Note sur la tuberculose des volailles. Recueil de Méd. Vét. Série VII, Vol. VIII (1891), p. 22. 6. Captot, GILBERT AND Rocsr. A contribution to the study of avian tuberculo- re mae in clinical veterinary medicine and surgery. (1900). (Translated by ollar. 7. CurisTiansen. Uber die Bedeutung der Gefliigeltuberkulose fir das Schwein. Zeitsch. fiir. Infek. Krankheiten, Bd. XIV (1913), p. 323. 8. Eperurern. Die Tuberculose der Papageien. Monatshefte fiir praktische Thierheilkunde, Bd. V (1894), S. 248. 9. Hastines, Haupin anp Brac. Avian tuberculosis. Journ. of Infect. Dis., Vol. XITI (1913), p. 1. 10. Lucer. Sur un symptome de la tuberculose chez la poule. Recueil de Méd. Vét., Série VII, Vol. VIII (1891), p. 172. 11. Marrucct. Die Hihnertuberculose. Zeitschr. fiir Hygiene, Bd. XI (1892), p. 445. 12. Moore anp Warp. Avian tuberculosis. Proc. Amer. Vet. Med. Asso., 1903, . 169. 13. Moors. The morbid anatomy and etiology of avian tuberculosis. Jour. of Med. Research, Vol. XI (1904), p. 521 (Bibliography). 14. Nocarp. Sur une tuberculose zoogléique des oiseaux de bassecour. Bul. et Memoires de la Soc. Centrale et Méd. Vét., 1885, p. 207. 15. Nocarp. Transmission de la tuberculose des poules et "homme. Veterinaire, Vol. III (1886), p. 658. 16. Prrnot. Investigations of diseases of poultry. Bulletin No. 64. Oregon Agric. Expt. Sta., 1900. 17. Sreumy. Tuberculosisin birds. Jour. of Compar. Med. and Vet. Arch., Vol. XI (1890), p. 317. 190 TUBERCULOSIS 18. Srraus er GAMALEIA. La tuberculose humaine, sa distinction de la tubercu- lose des oiseaux. Archiv. de Méd. Exper., Bd. III (1891), p. 457. ‘ 19. Straus er Wourrz. Sur la resistance des poules a la tuberculose par ingestion. Congress pour l étude de la tuberculose, 1888, p. 328. 20. Warp. Tuberculosis in fowls. Bulletin No. 161. California Agr. Exper. Station, 1904. 21. Werser. Review of the avian tuberculosis. Jour. of Compar. Med. and Vet. Arch., Vol. XIII (1892), p. 429. THE FOLLOWING BULLETINS ON TUBERCULOSIS HAVE BEEN ISSUED FROM THE VARIOUS STATE AGRICULTURAL EXPERIMENT STATIONS 1. Bane. The application of tuberculin in the suppression of bovine tuberculosis. Bulletin 41. Massachusetts. 1896. (A translation). 2. Bracu. The history of a tuberculous herd of cows. Bulletin 24. Storrs, Conn. 1902. 3. Brrrme. Bovine tuberculosis‘ Indiana. Bulletin 63. Ind. 1896. 4. Brewer. Tuberculosis. Bulletin 41. Utah. 1895. 5. Briscoz anp MacNeau. Tuberculosis of farm animals. Bulletin 149. Illinois. 1911. 6. Cary. Bovine tuberculosis. Bulletin 67. Alabama. 1895. 7. Conn. The relation of bovine tuberculosis to that of man and its significance in the dairy herd. - Bulletin 23. Storrs, Conn. 1902. 8. Drinwippiz. The relative virulence for the domestic animals of human and bovine tuberculosis. Bulletin 57. Kansas. 1899. 9. Dinwippie. The relative susceptibility of the domestic animals to the contagia of human and bovine tuberculosis. Bulletin 63. Kansas. 1900. 10. Farmer’s Butierin 473. U.S. Department of Agriculture, 1911. 11. Fiscuer. Bovine tuberculosis. Builletin 79. Kansas. 1898. 12. Gtover. Relation of bovine to human tuberculosis. Bulletin 66. Colorado. 1901. 13. Granez. Tuberculosis. Bulletin 133. Michigan. 1896. 14. Harpine, Smita snp Moors. The Bang method etc. Bulletin. Geneva, o = 1906. Harine ano Bett. The intradermal test for tuberculosis in cattle and hogs. Bulbstin 248. California. 1914. 16. Hitt anp Ricu. Bovine tuberculosis. Bulletin 42. Vermont. 1894. 17. Law. Tuberculosis in relation to animal industry and public health. Bulletin 65. (Cornell), New York. 1894. 18. Law. Experiments with tuberculin on non-tuberculous cows. Bulletin 82. (Cornell), New York. 1894. 19. Law. Tuberculosis in cattle and its control. Bulletin 150. (Cornell), New York. 1898. 20. Marsnaty. A study of normal temperatures and the tuberculin test. Bulletin 159. Michigan. 1898. 21. Marsuatu. Killing the tubercle bacilli in milk. Bulletin 173. Michigan. 22. Mayo. Some diseases of cattle, Texas itch, blackleg, tuberculosis, Texas fever. Bulletin 60. Kansas. 1897. 23. Mayo ano Kerr. Treatment of bovine tuberculosis. Bulletin 199. Vir- ginia. 1912. 24. Moors. The elimination of tubercle bacilli from infected cattle, and the con- trol of bovine tuberculosis and infected milk. Bulletin 299. (Cornell), New York. 1911. JOHNE’S DISEASE 191 25. Moorg. Bovine tuberculosis. Bulletin 226. (Cornell), N. Y. 1905. 26. Netson. On the use of Koch’s lymph in the diagnosis of tuberculosis. Report of the biologist. New Jersey. 1893. . 27. Netson. Experimental studies of the Koch test for tuberculosis. New Jersey, 1895. 28. NELSON. The suppression and prevention of tuberculosis of cattle and its relation to human consumption. Bulletin 118. New Jersey. 1896. 29. Nesom. Tuberculosis of cattle. Bulletin 50. S.C. 1900. _ 30. Paicu. History of tuberculosis in a college herd. Use of tuberculin in diagno- sis. Bulletin27. Massachusetts. 1894. 31. PEarson. Tuberculosis of cattle. Bulletin 29. Penn. 1894. 32. Reywnotps. Bovine tuberculosis. Bulletin 51. Minn. 1896. 33. RusseLy. Tuberculosis and the tuberculin test. Bulletin 40. Wisconsin. 34. Russett. The history of a tuberculous herd of cows. Bulletin 78. Wiscon- sin. 1899. 35. Russeiy. A lesson in bovine tuberculosis. Bulletin 114. Wisconsin. 1904. 36. Russe.t. Two ways of treating tuberculosis in herds. Bulletin 126. Wis- consin. 1905. 87. Russetu. The spread of tuberculosis through factory skim milk with sugges- tions as to its control. Bulletin 143. Wisconsin. 1907. 38. RusseLL anp Hastines. Bovine tuberculosis in Wisconsin. Bulletin 84. Wisconsin. 1901. 39. SraLKER AND Nixes. Investigation of bovine tuberculosis with special refer- ence to its existence in Iowa. Bulletin 39. Towa. 1895. 40. Tuorne. Bovine tuberculosis. Bulletin 108. Ohio. 1899. 41. Van Es. Bovine tuberculosis. Bulletin 77. North Dakota Agricultural Experiment Station. 1907.. 42. Wivu1amMson anp Emery. Tuberculosis and its prevention. Bulletin 117. N.C. 1895. 1. Report of the International Commission on the Control of Bovine Tuberculosis. Proceedings Am. Vet. Med. Asso., 1910, p. 90. 2. Report of the Royal Commission on Tuberculosis (Human and Bovine). Lon- don. 1907-1911. JOHNE’S DISEASE Synonyms. Specific paratuberculous enteritis; pseudo-tubercu- losis; chronic bovine pseudo-tuberculous enteritis; specific paratu- berculosis of cattle; enteritis paratuberculosis; bovis specifica; la diarrhée chronique du boeuf. Characterization. Johne's disease is an intestinal disorder caused by an acid-fast bacterium. It is characterized by a diarrhea, gradual emaciation and the presence of large numbers of acid-fast bacteria in the mucous membrane of the affected portions of the intestine. The small and large intestines and associated lymph glands are 192 JOHNE’S DISEASE involved. In addition to cattle, sheep, goats, deer, buffalo and possi- bly the horse are susceptible. History. Johne and Frothingham described a disease in 1895 in which the intestinal mucosa contained large numbers of acid-fast bacteria. They thought it was a case of tuberculosis in a cow due to the avian tubercle bacterium. In 1903 Markus called attention to its frequent occurrence in Holland. Since that time it has been recognized in Belgium, Switzerland, Denmark and England.