SS x \ ANS \\\ AN eS RS UUUUU! <* UUUU ‘oA 7. mi Ue - To JUUUUY ine JU q =f Eo } ) Gornell University Library Ithaca, New York j GEORGE FRANCIS ATKINSON BOTANICAL LIBRARY 1920 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 :/Awww.archive.org/details/cu31924003516923 PUTREFACTION AND INFECTION. PROFESSOR JOHN TYNDALL'S WORKS. Essays on the Floating Matter of the Air, in Relation to Putrefaction and Infection. 12mo. Cloth, $1.50. On Forms of Water, in Clouds, Rivers, Ice, and Gla- ciers. With 85 Illustrations, 12mo. Cloth, $1.50. Heat as a Mode of Motion. New edition. 12mo. loth, $2.50. On Sound; A Course of Eight Lectures delivered at the Royal Institution of Great Britain. Illustrated. 12mo. New edition. Cloth, $2.00. Fragments of Science for Unscientific People. 12mo. ew revised and enlarged edition. Cloth, $2.50. Light and Electricity. 12mo. Cloth, $1.25. Lessons in Electricity, 1715-16. 12mo. Cloth, $1.00. Hours of Exercise in the Alps. With Illustrations. 12mo. Cloth, $2.C0. a rs as @ Discoverer. A Memoir. 12mo. Cloth, Contributions to Molecular Physics in the Domain of Radiant Heat. $5.00. Six Lectures on Light. Delivered in America in 187278. With an Appendix and numerous Illus- trations. Cloth, $1.50. Farewell cif ha given to Professor Tyndall, at Del- monico’s, New York, February 4, 1878. Paper, 50 cents. Address delivered before the British Association, assembled at Belfast. Revised, with Additions, by the author, since the Delivery. 12mo. Paper, 50 cents. New York: D. Arrieton & Co., 1, 8, & 5 Bond St. ESSAYS ON THE FLOATING-MATTER OF THE AIR IN RELATION TO PUTREFACTION AND INFECTION. BY JOHN TYNDALL, F.R.S. (M.D. TUBINGEN). NEW YORK: D. APPLETON AND COMPANY, 1, 8, any 5 BOND STREET 1884. INTRODUCTORY NOTE. _— THE VIRTUAL TRIUMPH of the Antiseptic system of surgery, based as that system is on the recognition of living contagia as the agents of putrefaction, is of good augury as regards the receptivity of the public mind to new views respecting the nature of contagia generally. To the credit of English surgeons it stands recorded that, guided by their practical sagacity, they had adopted in their hospitals measures of amelioration which reduced, almost to a minimum, the rate of mor- tality arising from the ‘mortification’ of wounds. They had discovered the evils incident to ‘ dirt;’ and, by keeping dirt far away from them, they had saved innumerable lives, which would undoubtedly have suc- cumbed under conditions prevalent in many of the hospitals of continental Europe. In thus acting, English surgeons were, for the most part, ‘ wiser than they knew.’ Their knowledge, how- ever momentous in its practical applications, was still empirical knowledge. That dirt was fatal they had discovered ; but why it was fatal few of them knew. viii INTRODUCTORY NOTE. At this point Lister came forward with a scientific principle which rendered all plain. Dirt was fatal, not as dirt, but because it contained living germs which, as Schwann was the first to prove, are the cause of putrefaction. Lister extended the generalization of Schwann from dead matter to living matter, and by this apparently simple step revolutionized the art of surgery. He changed it, in fact, from an art into a science. ‘Listerism’ is sometimes spoken of as if it merely consisted in the application of carbolic acid spray; but no man of any breadth of vision will regard the subject thus. The antiseptic system had been enun- ciated, expounded, and illustrated, prior to the intro- duction of the spray. The spray is a mere offshoot of the system—elegant and effective it is true, but still a matter of detail. In company with my excellent friend Mr. John Simon, I once visited St. Bar- tholomew’s Hospital, and became acquainted, in its wards, with the practice of the late Mr. Callender. The antiseptic system was there as stringently applied as at King’s College. Immediately before his departure to America I spoke to Mr. Callender on this subject ; and he then told me expressly, that his aim and hope had been, not to introduce a new principle, but to simplify the methods of Lister. And yet Mr. Callen- der’s practice is sometimes spoken of as if it were, in principle, different from that of his eminent contem- porary. It is interesting, and indeed pathetic, to observe how long a discovery of priceless value to humanity INTRODUCTORY NOTE. ix may be hidden away, or rather lie openly revealed, before the final and apparently obvious step is taken towards its practical application. In 1837, Schwann clearly established the connexion between putrefaction and microscopic life; but thirty years had to elapse before Lister extended to wounds the researches of Schwann on dead flesh and animal infusions. Prior to Lister the possibility of some such extension had occurred to other minds. Penetrative men had seen that the germs which produce the putrefaction of meat might also act with fatal effect in the wards of a hospital. Thus, for example, in a paper read before the British Medical Association at Cambridge in 1864, Mr. Spencer Wells pointed out that the experiments of Pasteur, then recent, had ‘all a very important bearing upon the development of purulent infection, and the whole class of diseases most fatal in hospitals and other over- crowded places.’ Mr. Wells did not, as far as I know, introduce any systematic mode of combating the organ- isms whose power he so early recognised. But, I believe, in hardly any other department of surgery has the success of the antiseptic system been more con- spicuous and complete than in that particular sphere of practice in which Mr. Wells has won so great a name. A remark in the paper just referred to would seem to indicate that, in regard to the further possible influence of germs, the thoughts of Mr. Spencer Wells had passed beyond the bounds of pure surgical -prac- tice. ‘Their influence,’ he says, ‘ on the propagation of x INTRODUCTORY NOTE. epidemic and contagious diseases has yet to be made out.’ This shows that at the time here referred to the Germ Theory, in its wider medical sense, had begun to ferment in England. Two years, indeed, prior to the above occasion, and for the use of the same Association as that addressed by Mr. Wells, the late Dr. William Budd had drawn up a series of ‘Suggestions towards a Scheme for the Investigation of Epidemic and Epizootic Diseases,’ which strikingly illustrate the insight of a moan of genius, withdrawn from the stimulus of the metropolis, and working alone, at a time when the whole medical profession in England entertained views opposed to his. Budd states in succession, and with perfect clearness, the points which he considers most worthy of the attention of the Association. He re- commends inquiry as to the nature of the evidence alleged to prove the disease under investigation to be contagious or communicable. Whether such disease admits of being artificially propagated by inoculation or otherwise. Through what surface or surfaces the virus may be shown to enter the body, and to leave it, when the disease is taken in the natural way. Whether the disease is distinguished by eruptions external or internal. Whether it has a period of true incubation; and if so, what are the length and limits of that period. Whether one attack, as in smallpox and many other contagious diseases, preserves against future attacks. Whether in the case of human disease animals as well as man are susceptible, and if so, what animals. What, is the evidence, if any, as to the particular country or region in which the disease first appeared. What are INTRODUCTORY NOTE. xi its present geographical limits. Whether there is any evidence of its modern or recent introduction into countries previously exempt. How far any such disease may have been prevented from invading new countries, or from spreading from any particular centre, by measures directed against contagion. Above all, to determine what is the nature, and what the true value, of the evidence supposed to show that the specific poison of a contagious disease may originate spon- taneously, or be generated de novo. ‘What we most want to know,’ adds Budd, ‘in regard to this whole group of diseases is, where, and how, the specific poisons which cause them, breed and multiply,’ Budd’s own relation to the question here raised was distinct and, under the circumstances, impressive. ‘ After giving many years of time and thought to an examination of the evidence bearing on. this question,’ he comes to the conclusion ‘that there is no proof whatever’ that the poisons of specific contagious diseases ever originate spontaneously. ‘That the evidence on which the contrary conclusion is founded is negative only; that evidence of precisely the same order, only to all appeararice still more cogent, would prove animals and plants, even of large species, to originate spontaneously; that this evidence is therefore of no weight ; and, lastly, that all the really important facts point the other way, and tend to prove that these poisons (to use a term which is probably provisional only), like animals and plants, however they may have once originated, are only propagated now by the law of continuous succession.’ xii INTRODUCTORY NOTE. The word ‘ poisons,’ here provisionally employed, was a concession on Budd’s part to his weaker brethren ; for he, without a shade of doubt, considered the poison to be a real living seed. There was, I believe, but one physician of eminence in England who, at the time here referred to, shared this conviction, and who im- parted to Budd the incalculable force derived from the approbation and encouragement of a wise and cele- brated man. It gives me singular pleasure to write down here the name of the venerable Sir Thomas Watson, who lent to William Budd unfailing counte- nance and support, and who has lived to see that the views which commended themselves to his philosophic judgment are at the present moment advancing with resistless momentum among the members of the medical profession. It was far otherwise at the time to which we here refer. ‘Opinions like these,’ said Budd, ‘are no doubt, at present, those of a small minority. A very large, and by far the most influential school in this country—a school which probably embraces the great majority of medical practitioners, and the whole of the “sanitary public”—holds the exact contrary; and teaches that sundry of these poisons are constantly being generated de novo by the material conditions which surround us.’ Budd’s remark regarding the spontaneous generation of ‘animals and plants, even of large species,’ is both pregnant and pertinent. In reference to special and solitary outbreaks of contagious fever, I have frequently heard physicians of distinction affirm, without apparent misgiving, the ‘impossibility’ of importation from INTRODUCTORY NOTE. xiii without. On such occasions a reply, in the strict sense affirmed by William Budd, was always at hand; for I was able to adduce cases of solitary mushrooms, found upon out of the way Alpine slopes, to which the evidence would apply with greater force than to the cases on which the physicians referred to based their conclusions. With the atmosphere as a vehicle of universal intercommunication, it is hard to see any just warrant for the reliance of medical men upon the negative evidence stigmatized by Budd as valueless. It is, however, evidence by which many physicians are still influenced, and the effects of which it will probably require a generation of doctors, brought up under other conditions of culture and of practice, to wholly sweep away. These conditions are growing up around us, and their influence will be all-pervading before long. Never before was medicine manned and officered as it is now. To name here the workers at present engaged in the investigation of communicable diseases would be to extend beyond all reasonable limits this Introductory Note. On the old Baconian lines of observation and experiment the work is carried on. The intercommuni- cation of scientific thought plays here a most important part. It will probably have been noticed, that while physiologists and physicians in England and elsewhere were drawing copiously from the store of facts furnished by the researches of Pasteur, that admirable investigator long kept himself clear of physiology and medicine. There is, indeed, reason to believe that he was spurred on to his most recent achievements by the papers of xiv INTRODUCTORY NOTE. Burdon Sanderson, Koch, and others. The union of scientific minds is, or ought to be, organic. They are parts of the same body, in which every memter, under penalty of atrophy and decay, must discharge its due share of the duty imposed upon the whole. Of this ‘body,’ a short time since, England provided one of the healthiest limbs , but round that limb legislation has lately thrown a ligature, which threatens to damage its circulation and to divert its energies into foreign channels. In observational medicine one fine piece of work may be here referred to—the masterly inquiry of Dr. Thorne Thorne into the outbreak of typhoid fever at Caterham and Redhill. Hundreds were smitten by this epidemic, and many died. The qualities of mind illustrated in Dr. Thorne’s inquiry match those dis- played by William Budd in his memorable investigation of asimilar outbreak in Devonshire. Dr. Budd’s process was centrifugal—tracing from a single case in the village of North Tawton, the ravages of the fever far and wide. Dr. Thorne’s process was centripetal— tracing the epidemic backwards from the multitude of cases first: presented, to the single individual whose infected excreta, poured into the well at Caterham, were the cause of all. The Essays here presented to the reader belong to the A BC of the great subject touched upon in the foregoing Note. The two principal ones, namely, Essays II. and III., were prepared for the Royal Society, and are published in the ‘ Philosophical Trans- actions’ for 1876 and 1877. But, though written for INTRODUCTORY NOTE. xv that learned body, I sought to render their style and logic so clear as to render them accessible to any fairly cultivated mind. The Essays on ‘Fermentation’ and ‘Spontaneous Generation’ have already appeared else- where; while the first Essay, on ‘Dust and Disease, has been for some years before the public. It may be regarded as a kind of popular introduction to the more strenuous and original labours which follow it. The Essay most likely to try the reader’s patience is No. JII. On the whole, however, and particularly in its bearings on the Germ Theory of disease, it is probably the most important of all. The difficulties which sometimes beset the experimenter in these in- vestigations are best illustrated by this Essay. It shows, to my mind in a very impressive manner, the analogy of the spread of infection among organic in- fusions with its mode of propagation among human beings. The vital resistance of certain germs to heat is strikingly illustrated in the third Essay, one in- fusion being there proved to maintain its potentiality of life intact after eight hours’ continuous exposure to the temperature of boiling water. Under the plain guidance of the Germ Theory, it is however shown that an infusion of this stubborn character may be infallibly sterilized by discontinuous heating, in one hundredth part of the time requisite when the boiling is con- tinuous. Another question, to my mind of fundamental importance, is also disposed of in Essay III., where it is shown that the germs which exhibited the foregoing resistance are neither contained in the air, nor attached to the surface of the vessel, above the liquid, but that xvi INTRODUCTORY NOTE, they manifest their extraordinary vitality in the body of the liquid itself. On public sympathy the sanitary physician has mainly to rely for support, in a country where sanitary matters are left so much in the hands of the public itself as they are in England. But sympathy without cause—that is to say, without some basis of knowledge —is hardly to be expected. It is as a contribution to such knowledge that these Essays have been collected, and thrown into their present handy form. J. TYNDALL. Roya INSTITUTION: August 1881. CONTENTS. _—o——— ON DUST AND DISEASE. Experiments on Dusty Air : 5 . . . . The Germ Theory of Contagious Didensd . . oe Parasitic Diseases of Silkworms. Pasteur’s Researches ‘ ‘ Origin and Propagation of Contagious Matter . eo The Germ Theory applied to Surgery a . ‘ . . The Luminous Beam as a means of Research : A eas The Floating-Matter of the Air ‘ 3 . ‘ Dr. Bennett’s Experiments . . . . OPTICAL DEPORTMENT OF THE ATMOSPHERE IN RELATION TO PUTREFACTION AND INFEC- TION. § 1. Introduction : : : 2 : § 2. Method of Experiment ¢ : - : . . § 3. Deportment of Urine . . . : . : § 4. Mutton-Infusion . . . . . . . . § 5. Beef-Infusion . f . : . 7 oe § 6. Haddock-Infusion . a : . 7 a § 7. Turnip-Infusion . 3 . ‘ 2 . 5 rae § 8. Hay-Infusion . : . : 7 . . : . § 9. Infusion of Sole . ¥ ‘ . ‘ : . ae § 10. Liver-Infusion . : . a 5 § 11. Infusions of Hare, Rabbit, Pheasant, and Grouse a § 12. Infusions of Codfish, Turbot, Herring, and Mullet . § 13. Infusions of Fowland Kidney . . . § 14. Boiling by an Internal Source of Heat . ‘ ‘ § 15. Partial Discussion of the Results. . . . xviii § 16. § 17. § 18. § 19. § 20. § 21. § 22. § 23. § 24. § 26. § 26. § 27. CONTENTS. Suspended Particles in Air and Water; their relation to Bacteria . ‘ : . . o 25 Recent Experiments on a Heterogenesis . . . Experiments with Filtered Air . : Experiments with Calcined Air. Infusions withdrawn from Air . . . F ae The Germ Theory of Contagious Disease es 2 ‘ Experiments with Hermetically-sealed Vessels . is Conditions as to the Temperature and Strength of In- fusions . . Developmental Power e Infusions ond Solutions : Air. germs contrasted with Water-germs c Diffusion of Germs in the Air . Tray of one hundred Tubes 3 Some Experiments of Pasteur and ther Relation to Bae- terial Clouds . Note I. Action of Basteria apoN a Beara of Light Note II. Fluorescence of Infusions . 2 FURTHER RESEARCHES ON THE DEPORTMENT AND Cr tr Lr > wm Murra Cr Lr eh er Mr rar VITALITY OF PUTREFACTIVE ORGANISMS. . Introduction . : é . - . Experiments of Pasteur, Roberts ait Cohn i. . Hay-Infusions. Preliminary Experiments with Pipette- bulbs . Hay-Infusions. Hepariments with Cohn’s Walbes z . Hay-Infusions (in Closed Chambers) . . Desiccation of Germs. New Hay and old Hay-Infusions. Further Experiments with Closed Chambers . . . - : . . Experiments with Soaked Hay . Infusions of Fungi . . Infusions of Cucumber, Bectioot ie, : . New Experiments on Animal Infusions. Contradictory results . Infusions protected by ‘Glass Shades eontatning Ghlcinea, Air. F . Z : . . Further Precautions Gaalnet Tatestion, . Experiments in the Royal Gardens, Kew . Experiments on the Roof of the Royal Tdetitution. . Preliminary Experiments on the Resistance-limit of Germs to the temperature of Boiling Water 131 135 138 144 146 148 150 164 158 161 165 169 171 174 117 180 CONTENTS. xix § 17. Further Experiments on the Resistance-limit of Germs to the Boiling Temperature é 183 § 18. Change of Apparatus. New Hxperiments with Filtered Air . ;: : 188 § 19. Final proof that the Resistant Germs are extraned by the Infusion. Examples of Resistance both in Acid and Neutral Liquids " - + 194 20. Remarks on Acid, Neutral, and Alkatsaa Infusions - 203 § 21. Remarks on the Germs of Bacteria as eee from Bacteria themselves . . : - 205 § 22. Sterilization by discontinuous Heating . . 210 § 23. Mortality of Germs through defect of Oxygen produced by Exhaustion with the Sprengel Pump . : . 216 § 24. Mortality of Germs through defect of ca conse- quent on boiling the Infusion . . r + 221 § 25. Critical Review of the last two Sections . : 5 . 225 § 26. Mortality of Germs through excess of Oxygen . « » 227 § 27. Experiments on neutralized Urine . . ‘ . 228 § 28. Hermetically-sealed Flasks exposed to the Sun of the Alps . 7 3 . ‘ 5 . - » 231 § 29, Remarks on Hermetic Sealing : : : . 233 § 30. Experiments with Turnip-cheese Infusions 3 . . 234 FERMENTATION, AND ITS BEARINGS ON SURGERY AND MEDICINE . : : . . 287 SPONTANEOUS GENERATION . : ‘ - 277 APPENDIX. . . . . . . . ‘ . « 321 ON DUST AND DISEASE. —+oe—— I. Experiments on Dusty Air. Soar light, in passing through a dark room, reveals its track by illuminating the dust floating in the air. ‘The sun,’ says Daniel Culverwell, ‘discovers atomes, though they be invisible by candle-light, and makes them dance naked in his beams.’ In my researches on the decomposition of vapours by light, I was compelled to remove these ‘atomes’ and this dust. It was essential that the space con- taining the vapours should embrace no visible thing— that no substance capable of scattering light in the slightest sensible degree should, at the outset of an experiment, be found in the wide ‘ experimental tube’ in which the vapour was enclosed. For a long time I was troubled by the appearance there of floating matter, which, though invisible in diffuse daylight, was at once revealed by a powerfully condensed beam. Two U-tubes were placed in suc- cession in the path of the air, before it entered the liquid whose vapour was to be carried into the experi- mental tube. One of the U-tubes contained fragments ofmarble wetted with a strong solutionof caustic potash ; 2 THE FLOATING-MATTER OF THE AIR. the other, fragments of glass wetted with concentrated sulphuric acid which, while yielding no vapour of itsown, powerfully absorbs the aqueous vapour of the air. To my astonishment, the air of the Royal Institution, sent through these tubes at a rate sufficiently slow to dry it, and to remove its carbonic acid, carried into the experimental tube a considerable amount of mechani- cally suspended matter, which was illuminated when the beam passed through the tube. The effect was substantially the same when the air was permitted to bubble through the liquid acid, and through the solu- tion of potash. I tried to intercept this floating matter in various ways; and on October 5, 1868, prior to sending the air through the drying apparatus, it was carefully permitted to pass over the tip of a spirit-lamp flame. The floating matter then no longer appeared, having been burnt up by the flame. It was therefore organic matter. I was by no means prepared for this result; having previously thought that the dust of our air was, in great part, inorganic and non-combustible.! I had constructed a small gas-furnace, now much employed by chemists, containing a platinum tube, which could be heated to vivid redness.? Within this 1 According to an analysis kindly furnished to me by Dr. Percy, the dust collected from the walls of the British Museum contains fully 50 per cent. of inorganic matter. Ihave every confidence in the results of this distinguished chemist; they show that the jloat- ing dust of our rooms is, as it were, winnowed from the heavier matter. As bearing directly upon this point I may quote the fol- lowing passage from Pasteur: ‘Mais ici se présente une remarque: la poussiére que l’on trouve a la surface de tous les corps est soumise constamment 4 des courants d’air, qui doivent soulever-les parti- cules les plus légéres, au nombre desquelles se trouvent, sans donte, de préférence les corpuscules organisés, ceufs ou spores, moins lourds généralement que les particules minérales.’ 2 Pasteur was, I believe, the first to employ such a tube. DUST AND DISEASE. 3 tube was a roll of platinum gauze, which, while it permitted the air to pass through it, ensured the practical contact of the dust with the incandescent metal. The air of the laboratory was permitted to enter the experimental tube, sometimes through the cold, and sometimes through the heated, tube of plati- num. In the first column of the following fragment of a long table of results, the quantity of air operated on is expressed by the depression of the mercury gauge of the air-pump. In the second column the condition of the platinum tube is mentioned, and in the third the state of the air in the experimental tube. Quantity of air State of platinum tube State of experimental tube l5inches . Cold , a Full of particles. 30 yy , 7 Red-hot . 3 Optically empty. > The phrase ‘ optically empty’ shows that when the conditions of perfect combustion were present, the floating matter totally disappeared. In a cylindrical beam, which strongly illuminated the dust of the laboratory, I placed an ignited spirit- lamp. Mingling with the flame, and round its rim, were seen curious wreaths of darkness resembling an intensely black smoke. On placing the flame at some distance below the beam, the same dark masses stormed upwards. They were blacker than the blackest smoke ever seen issuing from the funnel of a steamer; and their resemblance to smoke was so perfect as to lead the most practised observer to conclude that the apparently pure flame of the alcohol lamp required but a beam of sufficient intensity to reveal its clouds of liberated carbon. But is the blackness smoke? This question pre- sented itself in a moment and was thus answered: A red-hot poker was placed underneath the beam: from 4 THE FLOATING-MATTER OF THE AIR. it the black wreaths also ascended. A large hydrogen flame was next employed, and it produced those whirl- ing masses of darkness, far more copiously than either. the spirit-flame or poker. Smoke was therefore out of the question.! What, then, was the blackness? It was simply that of stellar space; that is to say, blackness resulting from the absence from the track of the beam of al! matter competent to scatter its light. When the flame was placed below the beam the floating matter was destroyed in situ; and the air, freed from this matter, rose into the beam, jostled aside the illuminated par- ticles, and substituted for their light the darkness due to its own perfect transparency. Nothing could more forcibly illustrate the invisibility of the agent which renders all things visible. The beam crossed, unseen, the black chasm formed by the transparent air, while, at both sides of the gap, the thick-strewn particles shone out like a luminous solid under the powerful illumination. It is not, however, necessary to burn the particles to produce a stream of darkness. Without actual combustion, currents may be generated which shall displace the floating matter, and appear dark amid the surrounding brightness. I noticed this effect first. on placing a red-hot copper ball below the beam, and permitting it to remain there until its temperature had fallen below that of boiling water. The dark currents, though much enfeebled, were still produced. They may also be produced by a flask filled with hot water. 1 In none of the public rooms of the United States where I had the honour to lecture was this experiment made. The organic dust was too scanty. Certain rooms in England—the Brighton Pavilion, for example—also lack the necessary conditions. DUST AND DISEASE. 6 To study this effect a platinum wire was stretched transversely under the beam, the two ends of the wire being connected with the two poles of a voltaic battery. To regulate the strength of the current a rheostat was placed in the circuit. Beginning with a feeble current the temperature of the wire was gradually augmented ; but long before it reached the heat of ignition, a flat stream of air rose from it, which when looked at edge- ways appeared darker and sharper than one of the blackest lines of Fraunhofer in the purified spectrum. Right and left of this dark vertical band the floating matter rose upwards, bounding definitely the non- luminous stream of air. What is the explanation? Simply this: The hot wire rarefied the air in contact with it, but it did not equally lighten the floating matter. The convection current of pure air therefore passed upwards among the inert particles, dragging them after it right and left, but forming between them an impassable black partition. This elementary ex- periment enables us to render an account of the dark currents produced by bodies at a temperature below that of combustion. When the platinum wire is intensely heated, the floating matter is not only displaced, but destroyed. I stretched a wire about 4 inches long through the air of an ordinary glass shade resting on cotton-wool, which also surrounded the rim. The wire being raised to a white heat by an electric current, the air expanded, and some of it was forced through the cotton-wool. When the current was interrupted, and the air within the shade cooled, the returning air did not carry motes along with it, being filtered by the wool. At the beginning of this experiment the shade was charged with floating matter; at the end of half an hour it was optically empty. 6 THE FLOATING-MATTER OF THE AIR. On the wooden base of a cubical glass shade, a cubic foot in volume, upright supports were fixed, and from one support to the other 38 inches of platinum wire were stretched in four parallel lines. The ends of the platinum wire were soldered to two stout copper wires which passed through the base of the shade and could be connected with a battery. As in the last experiment the shade rested upon cotton-wool. A beam sent through the shade revealed the suspended matter. The platinum wire was then raised to white- ness. In five minutes there was a sensible diminution of the matter, and in ten minutes it was totally con- sumed. Oxygen, hydrogen, nitrogen, carbonic acid, so pre- pared as to exclude all floating particles, produce, when poured or blown into the beam, the darkness of stellar space. Coal-gas does the same. An ordinary glass shade, placed in the air with its mouth down- wards, permits the track of the beam to be seen crossing it. When coal-gas or hydrogen is allowed to enter the shade by a tube reaching to its top, the gas gradually fills the shade from above downwards. As soon as it occupies the space crossed by the beam, the luminous track is abolished. Lifting the shade so as to bring the common boundary of gas and air above the beam, the track flashes forth. After the shade is full, if it be inverted, the pure gas passes upwards like a black smoke among the illuminated particles. The Germ Theory of Contagious Disease. There is no respite to our contact with the floating matter of the air, We not only suffer from its mechanical irritation, but it is a growing belief that a portion of it lies at the root of a class of disorders DUST AND DISEASE, 7 most deadly toman. And what is this portion? It was some time ago the current belief that epidemic diseases generally were propagated by a kind of malaria, which consisted of organic matter in a state of motor. decay; that when such matter was taken into the body through the lungs, skin, or stomach, it had the power of spreading there the destroying process by which itself had been assailed. Such a power, it was alleged, was visibly exerted in the case of yeast. A little leaven was seen to leaven the whole lump—a mere speck of matter, in this supposed state of decomposition, being apparently competent to propagate indefinitely its own decay. Why should not a bit of rotten malaria within the human body act in a similar way? In 1836 a very unexpected reply was given to this question. In that year Cagniard de la Tour discovered the yeast- plant—a living organism, which when placed in a proper medium feeds, grows, and reproduces itself, and in this way carries on the process which we name fermentation. Here we have active life instead of motor-decay. By this discovery fermentation was con- nected with organic growth. Schwann, of Berlin, discovered the yeast-plant in- dependently about the same time; and in February, 1837, he also announced the important result, that when a decoction of meat is effectually screened from ordinary air, and supplied solely with calcined air, putrefaction never sets in. Putrefaction, therefore, he affirmed to be caused, not by the air, but by something in the air which could be destroyed by a sufficiently high temperature. The results of Schwann were confirmed by the independent experiments of Helmholtz, Ure, and Pasteur, while other methods, pursued by Schultze, and by Schroeder and Dusch, led to the same result. But as regards fermentation, the minds of chemists, 2 8 THE FLOATING-MATTER OF THE AIR. influenced probably by the great authority of Gay- Lussae, fell back upon the old notion of matter in a state of decay. It was not, they said, the living yeast-plant, but the dead or dying part of it, which, assailed by oxygen, produced the fermentation, But, as a matter of fact, when the plant is killed the ferment disappears. Mediate or immediate, the real ‘ferments’ are living organisms which find in fermentable substances their necessary food. Side by side with these researches and discoveries, and fortified by them and others, has run the germ theory of epidemic disease. The notion was expressed by Kircher, and favoured by Linnzus, that epidemic diseases may be due to germs which float in the at- mosphere, enter the body, and produce disturbance by the development within the body of parasitic life. The strength of this theory consists in the perfect parallelism of the phenomena of contagious disease with those of life. As a planted acorn gives birth to an oak, competent to produce a whole crop of acorns, each gifted with the power of reproducing its parent tree; and as thus from a single seedling a whole forest may spring ; so, it is contended, these epidemic diseases literally plant their seeds, grow, and shake abroad new germs, which, meeting in the human body their proper food and temperature, finally take possession of whole populations. There is nothing to my knowledge in pure chemistry which resembles the power of propaga- tion and self-multiplication possessed by the matter which produces epidemic disease. If you sow wheat you do not get barley; if you sow small-pox you do not get scarlet-fever, but small-pox indefinitely mul- tiplied, and nothing else. The matter of each con- tagious disease reproduces itself as rigidly as if it were (as Miss Nightingale puts it) dog or cat. DUST AND DISEASE. 9 Parasitic Diseases of Silkworms. Pasteur’s Researches. It is admitted on all hands that some diseases are the product of parasitic growth. Both in man and in lower creatures, the existence of such diseases has been demonstrated. I am enabled to lay before you an ac- count of an epidemic of this kind, thoroughly investi- gated and successfully combated by M. Pasteur. For fifteen years a plague had raged among the silkworms of France. They had sickened and died in multitudes, while those that succeeded in spinning their cocoons furnished only a fraction of the normal quantity of silk. In 1853 the silk culture of France produced a revenue of one hundred and thirty millions of franes. During the twenty previous years the revenue had doubled it- self, and no doubt was entertained as to its further augmentation. The weight of the cocoons produced in 1853 was 52,000,000 pounds; in 1865 it had fallen to 8,000,000, the fall entailing, in a single year, a loss of 100,000,000 francs. The country chiefly smitten by this calamity hap- pened to be that of the celebrated chemist Dumas, now perpetual secretary of the French Academy of Sciences. He turned to his friend, colleague, and pupil, Pasteur, and besought him, with an earnestness which the cir- cumstances rendered almost personal, to undertake the investigation of the malady. Pasteur at this time had never seen a silkworm, and he urged his inexperience in reply to his friend. But Dumas knew too well the qualities needed for such an inquiry to accept Pasteur’s reason for declining it. ‘Je mets,’ said he, ‘un prix extréme 4 voir votre attention fixée sur la question qui intéresse mon pauvre pays; la misére surpasse tout ce 10 THE FLOATING-MATTER OF THE AIR. que vous pouvez imaginer.’ Pamphlets about the plague had been showered upon the public, the mono- tony of waste paper being broken, at rare intervals, by a more or less useful publication. ‘ The Pharmacopeia of the Silkworm,’ wrote M. Cornalia in 1860, ‘is now as complicated as that of man. Gases, liquids, and solids have been laid under contribution. From eblorine to sulphurous acid, from nitric acid to rum, from sugar to sulphate of quinine,—all has been in- voked in behalf of this unhappy insect.’ The helpless cultivators, moreover, welcomed with ready trustfulness every new remedy, if only pressed upon them with sufficient hardihood. It seemed impossible to diminish their blind confidence in their blind guides. In 1863 the French Minister of Agriculture signed an agree- ment to pay 500,000 francs for the use of a remedy, which its promoter declared to be infallible. It was tried in twelve different departments of France, and found perfectly useless. In no single instance was it successful. It was under these circumstances that M. Pasteur, yielding to the entreaties of his friend, betook himself to Alais in the beginning of June, 1865. As regards silk husbandry, this was the most important department in France, and it was the most sorely smitten by the plague. The silkworm had been previously attacked by muscardine, a disease proved by Bassi to be caused by a vegetable parasite. This malady was propagated an- nually by the parasitic spores. Wafte:l by winds, they often sowed the disease in places far removed from the centre of infection. Muscardine is now said to be very rare, a deadlier malady having taken its place. This new disease is characterized by the black spots which cover the silkworms; hence the name pé. brine, first applied to the plague by M. de Quatrefages, DUST AND DISEASE. 11 and adopted by Pasteur. Pébrine declares itself in the stunted and unequal growth of the worms, in the languor of their movements, in their fastidiousness as regards food, and in their premature death. The course of discovery as regards the epidemic is this: In 1849 Guérin Méneville noticed in the blood of silkworms vibratory corpuscles, which he supposed from their motions to be endowed with independent life. Filippi, however, showed that the motion of the corpuscles was the well-known Brownian motion; but he committed the error of supposing the corpuscles to be normal to the life of the insect. Possessing the power of in- definite self-multiplication, they are really the cause of its mortality—the form and substance of its disease. This was well described by Cornalia; while Lebert and Frey subsequently found the corpuscles not only in the blood, but in all the tissues of the insect. Osimo, in 1857, discovered them in the eggs; and on this observation Vittadiani founded, in 1859, a prac- tical method of distinguishing healthy from diseased eggs. The test often proved fallacious, and it was never extensively applied. These living corpuscles first take possession of the intestinal canal, and spread thence throughout the body of the worm. They fill the silk cavities, the stricken insect often going automatically through the motions of spinning, without any material to work upon. Its organs, instead of being filled with the clear viscous liquid of the silk, are packed to distension by the corpuscles. On this feature of the plague Pasteur fixed his entire attention. The cycle of the silkworm’s life is briefly this: From the fertile egg comes the little worm, which grows, and casts its skin. This process of moulting is repeated two or three times at intervals during the life of the insect. After ' 12 THE FLOATING-MATTER OF THE AIR. the last moulting the worm climbs the brambles placed to receive it, and spins among them its cocoon. It passes thus into a chrysalis; the chrysalis becomes a moth, and the moth, when liberated, lays the eggs which form the starting-point of a new cycle. Now Pasteur proved that the plague-corpuscles might be incipient in the egg, and escape detection; they mignt also be germinal in the worm, and still baffle the microscope. But as the worm grows, the corpuscles grow also, becoming larger and more defined. In the aged chrysalis they are more pronounced than in the worm; while in the moth, if either the egg or the worm from which it comes should have been at all stricken, the corpuscles infallibly appear, offering no difficulty of detection. This was the first great point made out in 1865 by Pasteur. The Italian naturalists, as aforesaid, recommended the examination of the eggs before risking their incubation. Pasteur showed that both eggs and worms might be smitten, and still pass muster, the culture of such eggs or such worms being sure to entail disaster. He made the moth his starting- point in seeking to regenerate the race. Pasteur made his first communication on this sub- ject to the Academy of Sciences in September, 1865. It raised a cloud of criticism. Here, forsooth, was a chemist rashly quitting his proper métier and pre- suming to lay down the law for the physician and biologist on a subject which was eminently theirs. ‘On trouva étrange que je fusse si peu au courant de la question ; on m’opposa des travaux qui avaient paru depuis longtemps en Italie, dont les résultats montraient Vinutilité de mes efforts, et Vimpossibilité Warriver A un résultat pratique dans la direction que je m’étais engagé. Que mon ignorance fut grande au sujet des recherches sans nombre qui avaient paru depuis quinze DUST AND DISEASE. 13 années.’ Pasteur heard the buzz, but he continued his work. In choosing the eggs intended for incubation, the cultivators selected those produced in the successful ‘educations’ of the year. But they could not under- stand the frequent and often disastrous failures of their selected eggs; for they did not know, and nobody pricr to Pasteur was competent to tell them, that the finest cocoons may envelope doomed corpusculous moths. It, was not, however, easy to make the cultivators accept new guidance. To strike their imagination, and if possible determine their practice, Pasteur hit upon the expedient of prophecy. In 1866 he inspected, at St. Hippolyte-du-Fort, fourteen different parcels of eggs intended for incubation. Having examined a sufficient number of the moths which produced these eggs, he wrote out the prediction of what would occur in 1867, and placed the prophecy as a sealed letter in the hands of the Mayor of St. Hippolyte. In 1867 the cultivators communicated to the mayor their results. The letter of Pasteur was then opened and read, and it was found that in twelve out of fourteen cases there was absolute conformity between his pre- diction and the observed facts. Many of the groups had perished totally; the others had perished almost totally; and this was the prediction of Pasteur. In two out of the fourteen cases, instead of the prophesied destruction, half an average crop was obtained. Now, the parcels of eggs here referred to were considered healthy by their owners. They bad been hatched and tended in the firm hope that the labour expended on them would prove remunerative. The application of the moth-test for a few minutes in 1866 would have saved the labour and averted the disappointment. Two, additional parcels of eggs were at the same time sub- mitted to Pasteur. He pronounced them healthy ; and 14 THE FLOATING-MATTER OF THE AIR. his words were verified by the production of an excellent crop. Other cases of prophecy still more remarkable, because more circumstantial, are recorde.1 in Pasteur’s work on tne Diseases of Silkworms. Pasteur subjected the development of the corpuscles to a searching investigation, and followed out with admirable skill and completeness the various modes by which the plague was propagated. From moths per- fectly free from corpuscles he obtained healthy worms, and selecting 10, 20, 30, 50, as the case might be, he introduced into the worms the corpusculous matter. It was first permitted to accompany the food. Let us take a single example out of many. Rubbing up a stmoall corpusculous worm in water, he smeared the mixture over mulberry-leaves. Assuring himself that the leaves had been eaten, he watched the con- sequences from day to day. Side by side with the infected worms he reared their fellows, keeping them as much as possible out of the way of infection. These constituted his ‘lot témoin,—his standard of com- parison. On April 16, 1868, he thus infected thirty worms. Up to the 23rd they remained quite well. On the 25th they seemed well, but on that day cor- puscles were found in the intestines of two of them. On the 27th, or eleven days after the infected repast, two fresh worms were examined, and not only was the intestinal canal found in each case invaded, but the silk organ itself was charged with corpuscles. On the 28th the twenty-six remaining worms were covered by the black spots of pébrine. On the 30th the difference of size between the infected and non-infected worms was very striking, the sick worms being not more than two- thirds of the bulk of the healthy ones. On May 2 a worm which had just finished its fourth moulting was examined. Its whole body was so filled with the DUST AND DISEASE. 15 parasite as to excite astonishment that it could live. The disease advanced, the worms died and were ex- amined, and on May 11 only six out of the thirty remained. They were the strongest of the lot, but on being searched they also were found charged with cor- puscles. Not one of the thirty worms had escaped; a single meal had poisoned them all. The standard lot, on the contrary, spun their fine cocoons, two only of their moths being proved to contain any trace of the parasite, which had doubtless been introduced during the rearing of the worms. As his acquaintance with the subject increased, Pasteur’s desire for precision augmented, and he finally counted the growing number of corpuscles seen in the field of his microscope from day to day. After a con- tagious repast the number of worms containing the parasite gradually augmented until finally it became cent. per cent. The number of corpuscles would at the same time rise from 0 to 1, to 10, to 100, and some- times even to 1,000 or 1,500 in the field of his micro- scope. He then varied the mode of infection. He inoculated healthy worms with the corpusculous matter, and watched the consequent growth of the disease. He proved that the worms inoculate each other by the infliction of visible wounds with their claws. In various cases he washed the claws, and found corpuscles in the water. He demonstrated the spread of infection by the simple association of healthy and diseased worms. By their claws and their excrement, the diseased worms spread infection. It was no hypothetical infected medium-—no problematical pythogenic gas—that killed the worms, but a definite organism. The question of infection at a distance was also examined, and its occurrence demonstrated. As might be expected from Pasteur’s antecedents, the investigation was exhaustive. 16 THE FLOATING-MATTER OF THE AIR. the beauty of his manipulation finding a fit correlative in the clearness of his thought. The following quotation from Pasteur’s work clearly shows the relation in which his researches stand to the important question on which he was engaged : Place (he says) the most skilful cultivator, even the most expert microscopist, in presence of large cultivations which present the symptoms described in our experiments; his judgment will necessarily be erroneous if he confines him- self to the knowledge which preceded my researches. The worms will not present to him the slightest spot of pébrine ; the microscope will not reveal the existence of ccrpuscles ; the mortality of the worms will be null or insignificant; and the cocoons leave nothing to be desired. Our observer would, therefore, conclude without hesitation that the eggs produced will be good for incubation. The truth is, on the contrary, that all the worms of these fine crops have been poisoned ; that from the beginning they carried in them the germ of the malady ; ready to multiply itself beyond measure in the chrysalides and the moths, thence to pass into the eggs and smite with sterility the next generation. And what is the first cause of the evil concealed under so deceit- ful an exterior? In our experiments we can, so to speak, touch it with our fingers. It is entirely the effect of a single corpusculous repast; an effect more or less prompt according to the epoch of life of the worm that has eaten the poisoned food. Pasteur describes in detail his method of securing healthy eggs. It is nothing less than a mode of restor- ing to France her ancient silk husbandry. The justifi- cation of his work is to be found in the reports which reached him of the application and the suc- cess of his method, while editing his researches for final publication. In both France and Italy his method has been pursued with the most satisfactory results. But it was an up-hill fight which led to this: victory. DUST AND DISEASE, 17 ‘ Since the commencement,’ he says, ‘ of these researches, I have been constantly exposed to the most obstinate and unjust contradictions ; but I have made it a duty to leave no trace of these conflicts in this book.’ And in reference to parasitic diseases, generally, he uses the following weighty words: ‘I] est au pouvoir de ’homme de faire disparaitre de la surface du globe les maladies parasitaires, si, comme c’est ma conviction, la doctrine des générations spontanées est une chimére.’ Pasteur dwells upon the ease with which an island like Corsica might be absolutely isolated from the silk- worm epidemic. And with regard to other epidemics, Mr. John Simon describes an extraordinary case of in- sular exemption, for the ten years extending from 1851 to 1860. Of the 627 registration districts of England, one only had an entire escape from diseases which, in whole or in part, were prevalent in all the others: ‘In all the ten years it had not a single death by measles, nor a single death by small-pox, nor a single death by scarlet-fever. And why? Not because of its general sanitary merits, for it had an average amount of other evidence of unhealthiness. Doubtless, the reason of its escape was that it was insular. It was the district of the Scilly Isles; to which it was most improbable that any febrile contagion should come from without. And its escape is an approximative proof that, at least for those ten years, no contagium of measles, nor any contagium of scarlet-fever, nor any contagium of small- pox had arisen spontaneously within its limits. It may be added that there were only seven districts in England in which no death from diphtheria occurred, and that, of those seven, the district of the Scilly Isles was one. A second parasitic disease of silkworms, called in France la flacherie, co-existent with pébrine, but quite 18 THE FLOATING-MATTER OF THE AIR. distinct from it, has also been investigated by Pasteur. Enough, however, has been said to send those in- terested in these questions to the original volumes for further information. To one important practical point M. Pasteur, in a letter to myself, directs attention : Permettez-moi de terminer ces quelques lignes que je dois dicter, vaincu que je suis par la maladie, en vous faisant observer que vous rendriez service aux Colonies de la Grande- Bretagne en répandant la connaissance de ce livre, et des principes que j’¢tablis touchant la maladie des vers 4 soie. Beaucoup de ces colonies pourraient cultiver le mirier avec succés, et, en jetant les yeux sur mon ouvrage, vous vous convaincrez aisément qu'il est facile aujourd’hui, non- seulement d’éloigner la maladie régnante, mais en outre de donner aux récoltes de la soie une prospérité qu’elles n’ont jamais eue. Origin and Propagation of Contagious Matter. Prior to Pasteur, the most diverse and contradictory opinions were entertained as to the contagious character of pébrine; some stoutly affirmed it, others as stoutly denied it. But on one point all were agreed. ‘They believed in the existence of a deleterious medium, rendered epidemic by some occult and mysterious influence, to which was attributed the cause of the disease.’ Those acquainted with our medical literature will not fail to observe an instructive analogy here, We have on the one side accomplished writers, like Dr. Murchison, ascribing epidemic diseases to ‘ deleterious media’ which arise spontaneously in crowded hospitals and ill-smelling drains. According to them, the contagia of epidemic disease are formed de novo in a putrescent atmosphere. On the other side we have, writers like .. Dr. Budd, clear, vigorous, with well-defined ideas and DUST AND DISEASE, 19 methods of research, contending that the matter which produces epidemic disease comes always from a parent stock. It behaves as germinal matter, and they do not hesitate to regard it as such. They no more believe in the spontaneous generation of such diseases, than they do in the spontaneous generation of mice. Pasteur, for example, found that pébrine had been known for an in- definite time as a disease among silkworms. The de- velopment of it which he combated was merely the expansion of an already existing power—the bursting into open conflagration of*a previously smouldering fire. There is nothing surprising in this. For though epidemic disease requires a special contagium to pro- duce it, surrounding conditions must have a potent in- fluence on its development. Common seeds may be duly sown, but the conditions of temperature and mois- ture may be such as to restrict, or altogether prevent, the subsequent growth. Looked at, therefore, from the point: of view of the germ theory, the exceptional energy which epidemic disease from time to time exhibits is in harmony with the method of Nature. We sometimes hear diphtheria spoken of as if it were a new disease ; but Mr. Simon tells me that about three centuries ago tremendous epidemics of it raged in Spain (where it was named Garrotillo), and soon afterwards in Italy; and that since that time the disease has been well known to all successive generations of doctors. In or about 1758, for instance, Dr. Starr, of Liskeard, in a communication to the Royal Society, particularly described the disease, with all the symptoms which have recently again become familiar to us, but under the name of morbus strangula- torius, as then severely epidemic in Cornwall. This fact is the more interesting, as diphtheria, in its more modern reappearance, again showed predilection for that remote county. Many also believe that the Black Death, of 20 THE FLOATING-MATTER OF THE AIR. five centuries ago, has disappeared as mysteriously as it came; but Mr. Simon finds that it is believed to be prevalent at this hour in some of the north-western parts of India. Let me here state an item of my own experience. When I was at the Bel Alp in 1869, the English chaplain received letters informing him of the breaking out of scarlet-fever among his children. He lived, if I remember rightly, on the healthful eminence of Dart- moor, and it was difficult to imagine how scarlet-fever could have been wafted to the place. A drain ran close to his house, and on it his suspicions were manifestly fixed. Some of our medical writers would fortify him in this notion, and thus deflect him from the truth, while those of another, and, in my opinion, a wiser school, would deny to a drain, however foul, the power of generating de novo a specific disease. After close inquiry he recollected that a hobby-horse had been used both by his boy and by another who, a short time previously, had passed through scarlet-fever. Drains and cesspools, indeed, are by no means in such evil odour as they used to be. A fetid Thames and a low death-rate occur from time to time together in London. For, if the special matter or germs of epidemic disorder be not present, a corrupt atmosphere, dowever obnoxious otherwise, will not produce the disorder. But, if the germs be present, defective drains and cesspools become the potent distributors of disease and death. Corrupted air may promote an epidemic, but cannot produce it. On the other hand, through the transport of the special germ oz virus, disease may develop itself in regions where the drainage is good and the atmosphere pure. If you see a new thistle growing in your field, you feel sure that its seed has been wafted thither. Just as DUST AND DISEASE. 21 sure does it seem that the contagious matter of epi- demic disease has been sown in the place where it newly appears. With aclearness and conclusiveness not to be surpassed, Dr. William Budd hastraced such diseases from place to place; showing how they plant them- selves, at distinct points, among populations subjected to the same atmospheric influences, just as grains of corn might be carried in the pocket and sown. Hildebrand, to whose remarkable work, ‘ Du Typhus contagieux,’ Dr. de Mussy has directed my attention, gives the following striking case, both of the transport and the durability of the virus of scarlatina: ‘Un habit noir que j’avais en visitant une malade attaquée de scarlatine, et que je portai de Vienne en Podolie, sans Vavoir mis depuis plus d’un an et demi, me communiqua, dés que je fus arrivé, cette maladie contagieuse, que je répandis ensuite dans cette province, ot elle était jusqualors presque inconnue.’ Some years ago Dr. de Mussy himself was summoned to a country house in Surrey, to see a young lady who was suffering from a dropsy, evidently the consequence of scarlatina. The original disease, being of a very mild character, had been quite overlooked ; but circumstances were recorded which could leave no doubt upon the mind as to the nature and cause of the complaint. But then the question arose, How did the young lady catch the scarlatina? She had come there on a visit two months previously, and it was only after she had been a month in the house that she was taken ill. The housekeeper at length cleared up the mystery. The young lady, on her arrival, had expressed a wish to occupy a room in an isolated tower. Her desire was granted ; and in that room, six months previously, a visitor had been confined with an attack of scarlatina. The room had been swept and whitewashed, but the carpets had been permitted to remain. 22 THE FLOATING-MATTER OF THE AIR. Thousands of cases could probably be cited in which the disease has shown itself in this mysterious way, but where a strict examination has revealed its true parent- age and extraction. Is it, then, philosophical to take refuge in the fortuitous concourse of atoms as a cause of specific disease, merely because in special cases the parentage may be indistinct? Those best acquainted with atomic nature, and who are most ready to admit, as regards even higher things than this, the potentiali- ties of matter, will be the last to accept these rash hypotheses. The Germ Theory applied to Surgery. Not only medical but still more especially surgical science is now seeking light and guidance from this germ theory. Upon it the antiseptic system of Pro- fessor Lister of Edinburgh is founded. As already stated, the germ theory of putrefaction was started by Schwann; but the illustrations of this theory adduced by Professor Lister are of such public moment as not only to justify, but to render imperative, their intro- duction here. Schwann’s observations (says Professor Lister) did not receive the attention which they appeared to me to have deserved, The fermentation of sugar was generally allowed to be occasioned by the torila cerevisiae; but it was not admitted that putrefaction was due to an analogous agency. And yet the two cases present a very striking parallel. In each a stable chemical compound, sugar in the cne case, albumen in the other, undergoes extraordinary chemical changes under the influence of an excessively minute quantity of a substance which, regarded chemically, we should suppose inert. Asan example of this in the case of putrefaction, let us take a circumstance often witnessed in the treatment DUST AND DISEASE. 23 of large chronic abscesses. In order to guard against the access of atmospheric air, we used to draw off the matter by means of a canula and trocar, such as you see here, consist- ing of a silver tube with a sharp-pointed steel rod fitted into it, and projecting beyond it. The instrument, dipped in oil, was thrust into the cavity of the abscess, the trocar was withdrawn, and the pus flowed out through the canula, care being taken by gentle pressure over the part to prevent the possibility of regurgitation. The canula was then drawn out with due precaution against the reflux of air. This method was frequently successful as to its immediate object, the patient being relieved from the mass of the accumulated fluid, and experiencing no inconvenience from the operation. But the pus was pretty certain to reaccumulate in course of time, and it became necessary again and again to repeat the process. And unhappily there was no absolute security of immunity from bad consequences. However carefully the procedure was conducted, it sometimes happened, even though the puncture seemed healing by first intention, that feverish symptoms declared themselves in the course of the first or second day, and, on inspecting the seat of the abscess, the skin was perhaps seen to be red, implying the presence of some cause of irritation, while a rapid reaccumulation of the fluid was found to have occurred. Under these circum- stances, it became necessary to open the abscess by free incision, when a quantity, large in proportion to the size of the abscess, say, for example, a quart, of pus escaped, fetid” from putrefaction. Now, how had this change been brought about? Without the germ theory, I venture to say, no rational explanation of it could have been given. It must have been caused by the introduction of something from without. - Inflammation of the punctured wound, even sup- posing it to have occurred, would not explain the pheno- menon. For mere inflammation, whether acute or chronic, though it occasions the formation of pus, does not induce putrefaction. The pus originally evacuated was perfectly sweet, and we know nothing to account for the alteration in its quality but the influence of something derived from 24 THE FLOATING-MATTER OF THE AIR. the external world. And what could that something be? The dipping of the instrument in oil, and the subsequent precautions, prevented the entrance of oxygen. Or even if you allowed that a few atoms of the gas did enter, it would be an extraordinary assumption to make that these could in so short a time effect such changes in so large a mass of albuminous material. Besides, the pyogenic membrane is abundantly supplied with capillary vessels, through which arterial blood, rich in oxygen, is perpetually flowing; and there can be little doubt that the pus, before it was evacu- ated at all, was liable to any action which the element might be disposed to exert upon it. On the oxygen theory, then, the occurrence of putrefac- tion under these circumstances is quite inexplicable. But if you admit the germ theory, the difficulty vanishes at once. The canula and trocar having been lying exposed to the air, dust will have been deposited upon them, and will be present in the angle between the trocar and the silver tube, and in that protected situation will fail to be wiped off when the instrument is thrust through the tissues. Then when the trocar is withdrawn, some portions of this dust will naturally remain upon the margin of the canula, which is left projecting into the abscess, and nothing is more likely than that some particles may fail to be washed off by the stream of out-flowing pus, but may be dislodged when the tube is taken out, and left behind in the cavity. The germ theory tells us that these particles of dust will be pretty sure to contain the germs of putrefactive organisms, and if one such is left in the albuminous liquid, it will rapidly develop at the high temperature of the body, and account for all the phenomena. But striking as is the parallel] between putrefaction in this instance and the vinous fermentation, as regards the greatness of the effect produced, compared with the minuteness and the inertness, chemically speaking, of the cause, you will natu- rally desire further evidence of the similarity of the two processes. You can see with the microscope the torula of fermenting must or beer. Is there you may ask, any DUST AND DISEASE. 25 organism to be detected in the putrefying pus? Yes, gentle- men, there is. If any drop of the putrid matter is examined with a good glass, it is found to be teeming with myriads of minute jointed bodies, called vibrios, which indubitably pro- claim their vitality by the energy of their movements. It is not an affair of probability, but a fact, that the entire mass of that quart of pus has become peopled with living organisms as the result of the introduction of the canu'a and trocar; for the matter first let out was as free from vibrios as it was from putrefaction. If this be so, the greatness of the chemical changes that have taken place in the pus ceases to be surprising. We know that it is one of the chief pecu- liarities of living structures that they possess extraordinary powers of effecting chemical changes in materials in their vicinity, out of all proportion to their energy as mere chemical compounds. And we can hardly doubt that the animalcules which have been developed in the albuminous liquid, and have grown at its expense, must have altered its constitu- tion, just as we ourselves alter that of the materials on which we feed.! In antiseptic operations care is taken that every portion of tissue laid bare by the knife shall be de- fended from germs; that if they fall upon the wound they should be killed as they fall. With this in view he showers upon his exposed surfaces the spray of dilute carbolic acid, which is particularly deadly to the germs, and he surrounds the wound in the most careful manner with antiseptic bandages. To those accustomed to strict experiment it is manifest that we have a strict experimenter here—a man with a perfectly distinct object in view, which he pursues with never-tiring patience and unwavering faith. And the result, in his hospital practice, as described by himself, has been, that even in the midst of abominations too shocking to be mentioned here, and in the neighbourhood of 1 Introductory Lecture before the University of Edinburgh. 26 THE FLOATING-MATTER OF THE AIR. wards where death was rampant from pyzmia, erysipe- las, and hospital gangrene, he was able to keep his patients absolutely free from these terrible scourges. Let me here recommend to your attention Professor Lister’s ‘Introductory Lecture before the University of Edinburgh,’ which I have already quoted ; his paper on ‘The Effect of the Antiseptic System of Treacment on the Salubrity of a Surgical Hospital ;’ and the article in the ‘ British Medical Journal’ of January 14, 1871. If, instead of using carbolic acid spray, he could surround his wounds with properly filtered air, the re- sult would, he contends, be the same. In a room, where the germs not only float but cling to clothes and walls, this would be difficult, if not impossible. But surgery is acquainted with a class of wounds in which the blood is freely mixed with air that has passed through the lungs, and it is a most remarkable fact that such air does not produce putrefaction. Professor Lister, as far as I know, was the first to give a philosophical interpre- tation of this fact, which he describes and comments upon thus: I have explained to my own mind the remarkable fact that in simple fracture of the ribs, if the lung be punctured by a fragment, the blood effused into the pleural cavity, though freely mixed with air, undergoes no decomposition. The air is sometimes pumped into the pleural cavity in such abundance that, making its way through the wound in the pleura costalis, it inflates the cellular tissue of the whole body. Yet this occasions no alarm to the surgeon (although if the blood in the pleura were to putrefy, it would infallibly occasion dangerous suppurative pleurisy). Why air intro- duced into the pleural cavity through a wounded lung, should have such wholly different effects from that entering directly through a wound in the chest, was to me a complete mystery until I heard of the germ theory of putrefaction, when it at once occurred to me that it was only natural that air should DUST AND DISEASE. 27 be filtered of germs by the air-passages, one of whose offices is to arrest inhaled particles of dust, and prevent them from centering the air-cells. I shall have occasion to refer to this remarkable hypo- thesis farther on. The advocates of the germ theory, both of putrefac- tion and epidemic disease, hold that both arise, not from the air, but from something contained in the air. They hold, moreover, that this ‘something’ is not a vapour nor a gas, nor indeed a molecule of any kind, but a particle! The term ‘ particulate’ has been used by Mr. Simon in the Reports of the Medical Department of the Privy Council to describe this supposed constitu- tion of contagious matter; and Dr. Sanderson’s experi- ments render it in the highest degree probable, if they do not actually demonstrate, that the virus of small-pox is ‘ particulate.’ Definite knowledge upon this point is of exceeding importance, because in the treatment of par- ticles methods are available which it would be futile to apply to molecules. The Luminous Beam as a means of Research. My own interference with this great question, while sanctioned by eminent names, has been also an object of varied and ingenious attack. On this point I will only say that when angry feeling escapes trom behind the intellect, where it may be useful as an urging force, and places itself athwart the intellect, it is liable to 1 As regards size, there is probably no sharp line of division between molecules and particles; the one gradually shades into the other. But the distinction that I draw is this: the atom or the molecule, if free, is always part of a gas, the particle is never go, A particle is a bit of liquid or solid matter, formed by the aggregation of atoms or molecules. 28 THE FLOATING-MATTER OF THE AIR. produce all manner of delusions. Thus my censors, for the most part, have levelled their remarks against posi- tions which were never assumed, and against claims which were never made. The simple history of the matter is this: During the autumn of 1868 I was much occupied with the observations referred to at the be- ‘ ginning of this discourse, and in part described in the preceding article. For fifteen years it had been my habit to make use of floating dust to reveal the paths of luminous beams through the air; but until 1868 I did not intentionally reverse the process, and employ a luminous beam to reveal and examine the dust. Ina paper presented to the Royal Society in December, 1869, the observations which induced me to give more special attention to the question of spontaneous generation, and the germ theory of epidemic disease, are thus described : The Floating Matter of the Air. Prior to the discovery of the foregoing action (the chemical action of light upon vapours), and also during the experiments just referred to, the nature of my work compelled me to aim at obtaining experimental tubes abso- lutely clean upon the interior surface, and absolutely free within from suspended matter. Neither condition is, how- ever, easily attained. For however well the tubes might be washed and polished, and however bright and pure they might appear in ordi- nary daylight, the electric beam infallibly revealed signs and tokens of dirt. The air was always present, and it was sure to deposit some impurity, All chemical processes, not con- ducted in a vacuum, are open to this disturbance. When the experimental tube was exhausted, it exhibited no trace of floating matter, but on admitting the air through two U-tubes containing respectively caustic potash and sulphuric acid, a DUST AND DISEASE, 29 dust-cone more or less distinct was always revealed by the powerfully condensed electric beam. The floating motes resembled minute particles of liquid which had bien carried mechanically from the U-tubes into the experimental tube. Precautions were therefore taken to prevent any such transfer. They produced little or no miti- gation. I did not imagine, at the time, that the dust of the external air could find such free passage through the caustic potash and sulphuric acid. This, however, was the case; the motes really came from without. They also passed with freedom through a variety of ewthers and alcohols. In fact, it requires long-continued action on the part of an acid first to wet the motes and afterwards to destroy them. By care- fully passing the air through the flame of a spirit lamp, or through a platinum tube heated to bright redness, the floating matter was sensibly destroyed. It was therefore combustible, in other words, organic, matter. I tried to intercept it by a large respirator of cotton-woo'. Close pres- sure was necessary to render the wool effective. A plug of the wool, rammed pretty tightly into the tube through which the air passed, was finally found competent to hold back the motes. They appeared from time to time after- wards, and gave me much trouble ; but they were invariably traced in the end to some defect in the purifying apparatus —to some crack or flaw in the sealing-wax employed to render the tubes air-tight. Thus through proper care, but not with- out a great deal of searching out of disturbances, the expe- rimental tube, even when filled with air or vapour, contains nothing competent to scatter the light. The space within it has the aspect of an absolute vacuum. An experimental tube in this condition I call optically empty. The facts here forced upon my attention had a bear- ing too evident to be overlooked. The inability of air which had been filtered through cotton-wool to generate microscopic life, had been demonstrated by Schroeder and Pasteur: here the cause of its impotence was ren- 30 THE FLOATING-MATTER OF THE AIR. dered evident to the eye. The experiment proved that no sensible amount of light was scattered by the mole- cules of the air; that the scattered light always arose from suspended particles. The fact moreover that the re- moval of these abolished simultaneously the power of scattering light and of originating life, obviously de- tached the life-originating power from the air, and fixed it on something suspended in the air. Gases of all kinds passed with freedom through the plug of cotton- woul; hence the thing whose removal by the cotton- wool rendered the gas impotent, could not itself have been matter in the gaseous condition. It at once occurred to me that the retina, protected as it was, in these experiments, from all extraneous light, might be converted into a new and powerful instrument of demon- stration in relation to the germ theory. The observations just described also revealed the danger incurred in experiments of this nature; showing that without an amount of care far beyond that hitherto bestowed upon them, such experiments left the door open to errors of the gravest description. It was especially manifest that the chemical method employed by Schultze in his experiments, and so often resorted to since, might lead to the most erroneous conse- quences; that neither acids nor alkalies had the power of rapid destruction hitherto ascribed to them. In short, the employment of the luminous beam rendered evident the cause of success in experiments rigidly con- ducted like those of Pasteur; while it made equally evident the certainty of failure in experiments less severely carried out. DUST’ AND DISEASE. 31 Dr. Bennett's Kaperiments. I do not wish to leave an assertion of this kind with- out proof or illustration. Take, then, the well-conceived experiments of Dr. Hughes Bennett, described before the Royal Society of Surgeons in Edinburgh on January 17, 1868.1 Into flasks containing decoctions of liquor- ice-root, hay, or tea, Mr. Bennett, by an ingenious method, forced air. The air was driven through two U-tubes, the one containing a solution of caustic potash, the other sulphuric acid. ‘All the bent tubes were filled with fragments of pumice-stone to ‘break up the air, so as to prevent the possibility of any germs passing through in the centre of bubbles.’ The air also passed through a Liebig’s bulb containing sulphuric acid, and also through a bulb containing gun-cotton. It was only natural for Dr. Bennett to believe that his ‘ bent tubes’ entirely cut off the germs. Previous to the observations just referred to, I also believed in their efficacy. But these observations destroy any such notion. The gun-cotton, moreover, will fail to arrest the whole of the floating matter, unless it is tightly packed, and there is no indication in Dr. Bennett's memoir that it was so packed. On the whole, I should infer, from the mere inspection of Dr. Bennett’s appa- ratus, the very results which he has described—a retar- dation of the development of life, a total absence of it in some cases, and its presence in others. In his first series of experiments, eight flasks were fed with sifted air, and five with common air. In ten or twelve days all the five had fungi in them; whilst: it required from four to nine months to develop fungi in the others. In one of the eight, moreover, 1 British Medical Journal, 13, pt. ii. 1868. 3 32 THE FLOATING-MATTER OF THE ATR, even after this interval no fungi appeared. Ina second series of experiments there was a similar exception. In a third series the cork stoppers used in the first and second series were abandoned, and glass stoppers em- ployed. Flasks containing decoctions of tea, beef, and hay were filled with common air, and other flasks with sifted air. In every one of the former fungi appeared and in not one of the latter. These experiments simply ruin the doctrine that Dr. Bennett finally espouses. In all these negative cases, the air was forced through the bent tubes and bulb into the boiling-hot infusion. Dr. Bennett made a fourth series of experiments, in which, previous to forcing in the air, he permitted the flasks to cool. Into four bottles thus treated he forced pre- pared air, and after a time found fungi in all of them. What is his conclusion? Not that the boiling-hot liquid, employed in his first experiments, had destroyed such germs as had run the gauntlet of his apparatus ; but that air, which, previous to being sealed up, had been exposed to a temperature of 212°, is too rare to support life. This conclusion is so remarkable that it ought to be stated in Dr. Bennett’s own words. ‘It may be easily conceived that air subjected to a boiling temperature is so expanded as scarcely to merit the name of air, and that it is more or less unfit for the pur- pose of sustaining animal or vegetable life.’ Numerical data are attainable here, but they are unnecessary. As a matter of fact, I live and flourish for a considerable portion of each year in a medium of less density than that which Dr. Bennett describes as scarcely meriting the name of air. The inhabitants of the higher Alpine chalets, with their flocks and herds, and the grasses which support these, do the same; while the chamois rears its kids in air rarer still. DUST AND DISEASE. 33 Insect’ life, moreover, is sometimes exhibited with monstrous prodigality at Alpine heights. In a fifth series of experiments sixteen bottles were filled with infusions. Into four of them, while cold, ordinary unheated and unsifted air was pumped, and in these fungi were developed. Into four other bottles, containing a boiling infusion, ordinary air was also pumped-——no fungi were here developed. Into four other bottles containing an infusion which had been boiled and permitted to cool, sifted air was pumped —no fungi were developed. Finally, into four bottles containing a boiling infusion sifted air was pumped— no fungi were developed. Only, therefore, in the four cases where the infusions were cold infusions, and the air ordinary air, did fungi appear. Dr. Bennett does not draw from his experiments the conclusion to which they so obviously point. On the contrary, he founds on them a defence of the doctrine of spontaneous generation, and a general theory of sponta- neous development. So strongly was he impressed with the idea that the germs could not possibly pass through his potash and sulphuric acid tubes, that the appear- ance of fungi, even in a small minority of cases, where the air had been sent through these tubes, was to him conclusive evidence of the spontaneous origin of such fungi. And he accounts for the absence of life in many of his experiments by an explanation which will not bear a moment’s criticism. But knowing, as we now do, that organic particles may pass unscathed through alkalies and acids, the results of Dr. Bennett are pre- cisely what ought under the circumstances to be ex- pected. Indeed, their harmony with the conditions now revealed is a proof of the honesty and accuracy with which they were executed. The caution exercised by Pasteur both in the exe- 34 THE FLOATING-MATTER OF THE AIR. cution of his experiments, and in the reasoning based upon them, is perfectly evident to those who, through the practice of severe experimental inquiry, have ren- dered themselves competent to judge of good experi- mental work. He found germs in the mercury used to isolate his air. He was never sure that they did not cling to the instruments he employed, or to his own person. Thus when he opened his hermetically-sealed flasks upon the Mer de Glace, he had his eye upon the file used to detach the drawn-out necks of his bottles; and he was careful to stand to leeward when each flask was opened. Using these precautions, he found the glacier air incompetent, in nineteen cases out of twenty, to generate life ; while similar flasks, opened amid the vegetation of the lowlands, were soon crowded with living things. M. Pouchet repeated Pasteur’s experi- ments in the Pyrenees, adopting the precaution of holding his flasks above his head, and obtaining a different result. Now great care would be needed to render this procedure a real precaution. The luminous beam at once shows us its possible effect. Let smoking brown paper be placed at the open mouth of a glass shade, so that the smoke shall ascend and fill the shade. A beam sent through the shade forms a bright track through the smoke. When the closed fist is placed underneath the shade, a vertical wind of surprising violence, considering the small elevation of temperature, rises from the hand, displacing by comparatively dark ‘air the illuminated smoke. Unless special care were taken such a wind would rise from M. Pouchet’s body as he held his flasks above his head, and thus the pre- caution of Pasteur, of not coming between the wind and the flask, would be annulled. Let me now direct attention to another result of Pasteur, the cause und significance of which are at once DUST AND DISEASE. 35 revealed by the luminous beam. He prepared twenty- one flasks, each containing a decoction of yeast, filtered and clear. He boiled the decoction so as to destroy whatever germs it might contain, and while the space above the liquid was filled with pure steam, he sealed his flasks with a blow-pipe. He opened ten of them in the deep, damp caves of the Paris Observatory, and eleven of them in the courtyard of the establishment. Of the former, one only showed signs of life subse- quently. In nine out of the ten flasks no organisms of any kind were developed. In all the others organisms speedily appeared. Now here is an experiment conducted in Paris, on which we can throw light in London. Causing our luminous beam to pass through a large flask filled with the air of this room, and charged with its germs and its dust, the beam is seen crossing the flask from side toside. But here is another similar flask, which cuts a clear gap out of the beam. It is filled with wn- jiltered air, and still no trace of the beam is visible. Why? By pure accident I stumbled on this flask in our apparatus room, where it had remained quiet for some time. Acting upon this obvious suggestion, I set aside three other flasks, filled, in the first instance, with mote-laden air. They are now optically empty. Our former experiments proved that the life-producing particles attach themselves to the fibres of cotton-wool. In the present experiment the motes have been brought by gentle air-currents, established by slight differences of temperature within our closed vessels, into contact with the interior surface, to which they adhere. The air of these flasks has deposited its dust, germs and all, and is practically free from suspended matter. I had a chamber erected, the lower half of whichis of wood, its upper half being enclosed by four glazed 36 THE FLOATING-MATTER OF THE AIR. window-frames. It tapers to a truncated cone at the top. It measures in plan 3 ft. by 2 ft. 6 in., and its height is 5 ft. 10 in. On February 6 it was closed, every crevice that could admit dust, or cause displace- ment of the air, being carefully pasted over with paper. The electric beam at first revealed the dust within the chamber as it did in the air of the laboratory. The chamber was examined almost daily; a perceptible diminution of the floating matter being noticed as time advanced. At the end of a week the chamber was optically empty, exhibiting no trace of matter competent to scatter the light. Such must have been the case in the stagnant caves of the Paris Observatory. Were our electric beam sent through the air of these caves, its track would, doubtless, save from aqueous haze, be invi- sible ; thus showing the indissoluble association of the scattering of light by air and its power to generate life. I will now turn to what seems to me a more interest- ing application of the luminous beam than any hitherto described. My reference to Professor Lister’s interpre- tation of the fact, that air which has passed through the lungs cannot produce putrefaction, is fresh in your memories. ‘Why air, said he, ‘introduced into the pleural cavity, through a wounded lung, should have such wholly different effects from that entering through a permanently open wound, penetrating from without, was to me a complete mystery, till I heard of the germ theory of putrefaction, when it at once occurred to me that it was only natural that the air should be filtered of germs by the air passages, one of whose offices is to arrest inhaled particles of dust, and prevent them from entering the air-cells.’ Here is a surmise which bears the stamp of genius but which needs verification. If, for the words ‘it is only natural’ we were authorized to write ‘it is per- DUST AND DISEASE. 37 fectly certain,’ the demonstration would be complete. Such demonstration is furnished by experiments with a beam of light. One evening, towards the close of 1869, while pouring various pure gases across the dusty track of a luminous beam, the thought occurred to me of using my breath instead of the gases. I then noticed, for the first time, the extraordinary darkness produced by the expired air, towards the end of the expiration. Permit me to repeat the experiment in your presence. I fill my lungs with ordinary air and breathe through a glass tube across the beam. The condensation of the aqueous vapour of the breath is shown by the formation of a luminous white cloud of delicate texture. We abolish this cloud by drying the breath previous to its entering the beam; or, still more simply, by warming the glass tube. The luminous track of the beam is for atime uninterrupted by the breath, because the dust returning from the lungs makes good, in great part, the particles displaced. After a time, however, an obscure disk appears in the beam, the darkness of which in- creases, until finally, towards the end of the expiration, the beam is, as it were, pierced by an intensely black hole, in which no particles whatever can be discerned. The deeper air of ihe lungs is thus proved to be abso- lutely free from suspended matter. It is therefore in the precise condition required by Professor Lister’s explanation. This experiment may be repeated any number of times with the same result. I think it must be regarded as a crowning piece of evidence both of the correctness of Professor Lister’s views and of the impo- tence, as regards vital development, of optically pure air.! 1 Dr. Burdon Sanderson draws attention to the important observation of Brauell, which shows that the contagium of a preg- nant animal, suffering from splenic fever, is not found in the blood of the foetus ; the placental apparatus acting as a filter, and holding back the infective particles. 38 THE FLOATING-MATTER OF THE AIR.. The foregoing essay, as far as it relates to the theory which ascribes epidemic disease to the develop- ment of low parasitic life within the human life, was embodied in a discourse delivered before the Royal Institution in January 1870. In June 1871, after a brief reference to the polarization of light by cloudy matter, I ventured to recur to the subject in these terms: What is the practical use of these curiosities ? If we exclude the interest attached to the observation of new facts, and the enhancement of that interest through the knowledge that facts often become the exponents of laws, these curiosities are in themselves worth little. They will not enable us to add to our stock of food, or drink, or clothes, or jewellery. But though thus shorn of all usefulness in themselves, they may, by carrying thought into places which it would not otherwise have entered, become the antecedents of practical conse- quences. In looking, for example, at our illuminated dust, we may ask ourselves what it is. How does it act, not upon a beam of light, but upon our own bodies ? The question then assumes a practical character. We find on examination that this dust is mainly organic matter—in part living, in partdead. There are among it particles of ground straw, torn rays, smoke, the pollen of flowers, the spores of fungi, and the germs of other things. But what have they to do with the animal economy? Let me give you an illustration to which my attention has been lately drawn by Mr. George Henry Lewes, who writes to me thus: ‘I wish to direct your attention to the experiments of Von Recklingshausen, should you happen not to know them. They are striking confirmations of what you say of dust and disease. Last spring, when I was at his laboratory in Wurzburg, I examined with him blood that had been three weeks, a month, and five weeks, out DUST AND DISEASE, 39 of the body, preserved in little porcelain cups under glass shades. This blood was living and growing. Not only were the Amcba-like movements of the white corpuscles present, but there were abundant evidences of the growth and development of the corpuscles. I also saw a frog’s heart still pulsating which had been removed from the body (I forget how many days, but certainly more than a week). There were other examples of the same persistent vitality, or absence of putrefaction. Von Recklingshausen did not attribute this to the absence of germs—germs were not mentioned by him; but when I asked him how he represented the thing to himself, he said the whole mystery of his operation consisted in keeping the blood free from dirt. The instruments employed were raised to a red heat just before use; the thread was silver thread and was similarly treated; and the porcelain cups, though not kept free from air, were kept free from currents. He said he often had failures, and these he attributed to particles of dust having escaped his precautions.’ Professor Lister, who has founded upon the removal or destruction of this ‘dirt’ momentous improvements in surgery, tells us the effect of its introduction into the blood of wounds. The blood putrefies and he- comes fetid; and when you examine more closely what putrefaction means, you find the putrefying substance swarming with infusorial life, the germs of which have been derived from the atmospheric dust. We are now assuredly in the midst of practical mat- ters; and with your permission I will refer once more to a question which has recently occupied a good deal of public attention. As regards the lowest forms of life, the world is divided, and has for a long time been divided, into two parties, the one affirming that we have only to submit absolutely dead matter to certain 40 THE FLOATING-MATTER OF THE AIR. physical conditions, to evolve from it living things; the other (without wishing to set bounds to the power of matter) affirming that, in our duy, life has never been found to arise independently of pre-existing life. I- belong to the party which claims life as a derivative of life. The question has two factors—the evidence, and the mind that judges of the evidence; and it may be purely a mental set or bias on my part that causes me, throughout this long discussion, to see, on the one side, dubious facts and defective logic, and on the other side firm reasoning and a knowledge of what rigid experi- mental inquiry demands. But, judged of practically, what, again, has the question of Spontaneous Genera- tion to do with us? Let us see. There are numerous diseases of men and animals that are demonstrably the products of parasitic life, and such diseases may take the most terrible epidemic forms, as in the case of the silkworms of France, referred to at an earlier part of this essay. Now it is in the highest degree important to know whether the parasites in question are spontaneously developed, or whether they have been wafted from with- out to those afflicted with the disease. The means of prevention, if not of cure, would be widely different in the two cases. But this is not all. Besides these universally ad- “mitted cases, there is the broad theory, now broached and daily growing in strength and clearness—daily, in- deed, gaining more and more of assent from the most successful workers and profound thinkers of the medical profession itself—the theory, namely, that contagious disease, generally, is of this parasitic character. Had I any cause to regret having introduced this theory to your notice more than a year ago, that regret should now be expressed. I would certainly renounce in your presence whatever leaning towards the germ theory DUST AND DISEASE, 4l my words might then have betrayed. But since the time referred to nothing has occurred to shake my conviction of the truth of the theory. Let me briefly state the grounds on which its supporters rely. From their respective viruses you may plant typhoid fever, scarlatina, or small-pox. What is the crop that arises from this husbandry? As surely as a thistle rises from a thistle seed, as surely as the fig comes from the fig, the grape from the grape, the thorn from the thorn, so surely does the typhoid virus increase and multiply into typhoid fever, the scarlatina virus into scarlatina, the small-pox virus into small-pox. What is the con- clusion that suggests itself here? It is this: That the thing which we vaguely call a virus is to all intents and purposes a seed. Excluding the notion of vitality, in the whole range of chemical science you cannot point to an action which illustrates this perfect parallelism with the phenomena of life—this demonstrated power of self-multiplication and repro- duction. The germ theory alone accounts for the phenomena. In cases of epidemic disease, it is not on bad air or foul drains that the attention of the physician of the future will primarily be fixed, but upon disease germs, which no bad air or foul drains can create, but which may be pushed by foul air into virulent energy of repro- duction. You may think I am treading on dangerous ground, that I am putting forth views that may interfere with salutary practice. No such thing. If you wish to learn the impotence of medical practice in dealing with contagious diseases, you have only to refer to the Har- veian oration for 1871, by Sir William Gull. Such dis. eases defy the physician. They must run their course, and the utmost that can be done for them is careful nursing. And this, though I do not specially insist 42 THE FLOATING-MATTER OF THE AIR. upon it, would favour the idea of their vital origin. For if the seeds of contagious disease be themselves living things, it may be difficult to destroy either them or their progeny, without involving their living habitat in the same destruction. It has been said, and it is sure to be repeated, that I am quitting my own métier, in speaking of these things. Not so. Iam dealing with a question on which minds accustomed to weigh the value of experimental evidence are alone competent to decide, and regarding which, in its present condition, minds so trained are as capable of forming an opinion as regarding the pheno- mena of magnetism or radiant heat. ‘ The germ theory of disease,’ it has been said, * appertains to the biologist and the physician.’ Where, I would ask in reply, is the biologist or physician, whose researches, in connection with this subject, could for one instant be compared to those of the chemist Pasteur? It is not the philosophic members of the medical profession who are dull to the reception of truth not originated within the pale of the profession itself. I cannot better conclude this portion of my story than by reading to you an extract froma letter addressed to me some time ago by Dr. William Budd, of Clifton, to whose insight and energy the town of Bristol owes so much in the way of sanitary improve- ment. ‘As to the germ theory itself, writes Dr. Budd, ‘that is a matter on which I have long since made up my mind. From the day when I first began to think of these subjects I have never had a doubt that the specific cause of contagious fevers must be living organisms. ‘It is impossible, in fact, to make any statement bearing upon the essence or distinctive characters of these fevers, without using terms which are of all others the most distinctive of life Take up the writings of DUST AND DISEASE. 43 the most violent opponent of the germ theory, and, ten to one, you will find them full of such terms as “ pro- pagation,” “ self- propagation,” “ reproduction,” “ self- multiplication,” and soon. Try as he may—if he has anything to say of those diseases which is characteristic of them—he eannot evade the use of these terms, or the exact equivalents to them. While perfectly applicable to living things, these terms express qualities which are not only inapplicable to common chemical agents, but, as far as I can see, actually inconceivable of them.’ JPTICAL DEPORTMENT OF THE ATMOSPHERE IN RELATION TO PUTREFACTION AND INFECTION.’ a iT; § 1. Introduction. AN inquiry into the decomposition of vapours by light, begun in 1868 and continued in 1869,? in which it was necessary to employ optically pure air, led me to experi- ment on the floating matter of the atmosphere. A brief section of a paper published in the Philosophical Transactions for 18705 is devoted to this subject. I at that time found that the air of London rooms, which is always thick with motes, and also with matter too fine to be described as motes, after it had been filtered by passing it through densely packed cotton- wool, or calcined by passing it through a red-hot pla- tinum-tube containing a bundle of red-hot platinum wires, or by carefully leading it over the top of a spirit- lamp flame, showed, when examined by a concentrated luminous beam, no trace of mechanically suspended matter. The particular portion of space occupied by such a beam was not to be distinguished from adjacent, space. The purely gaseous portion of our atmosphere was thus shown to be incompetent to scatter light. 1 Philosophical Transactions, Part L, 1876. 2 Proc. Roy. Soe. vol. xvii. 3 Vol. clx. p. 337, 16 THE FLOATING-MATTER OF THE AIR, I subsequently found that, to render the air thus optically pure, it, was only necessary to leave it to itself for a sufficient time in a small closed chamber, or in a suitably closed vessel. The floating matter gradually attached itself to the top and sides, or sank to the bottom, leaving behind it air possessing no scattering power. Sent through such air, the most concentrated beam failed to render its track visible. I mention ‘top’ and ‘sides,’ as well as ‘ bottom,’ because gravity is not the only agent, perbaps not even the principal agent, concerned in the removal of the floating matter. It is practically impossible to sur- round a closed vessel by an absolutely uniform tempera- ture; and where differences of temperature, however small, exist, air-currents will be established. By such gentle currents the floating particles are gradually brought into contact with all the surrounding surfaces. To these they adhere, and, no new supply being ad- mitted, the suspended matter finally disappears from the air altogether. The striking parallelism of these results with those obtained in the excellent researches of Schwann,} Schroeder and Dusch,? Schroeder himself,? and Pasteur‘ in regard to the question of ‘spontaneous generation,’ caused me to conclude that the power of scattering light, and the power of producing life, by atmospheric air would be found to go hand in hand. This conclusion was strengthened by an experiment easily made and of high significance in relation to this question. It had been pointed out by Professor Lister ® 1 Poge. Ann. 1837, vol. xli. p. 184. 2 Ann. der Pharmacie, vol. lxxxix. p. 232. 3 Ibid. vol. cix. p. 35. 4 Ann. de Chim. ct de Phys. 3rd series, vol. Ixiv. p. 83. * Introductory Lecture before the University of Edinburgh. PUTREFACTION AND INFECTION. 47 that air which has passed through the lungs is known to have lost its power of causing putrefaction. Such air may mix freely with the blood of an internal wound without risk of mischief; and that truly great scientific surgeon had the penetration to ascribe this immunity from danger to the filtering power of the lungs. Prior to my becoming acquainted with this hypothesis in 1869, I had demonstrated its accuracy in the following manner.! Condensing in a dark room, and in dusty air, a Fig. 1, powerful beam of light, and breathing through a glass tube (the tube actually employed was a lamp-glass, ren- dered warm in a flame to prevent condensation of the breath) across the focus, a diminution of the scattered light was first observed. But towards the end of the expiration the white track of the beam was broken by a perfectly black gap, the blackness being due to the total absence from the expired air of any matter com- petent to scatter light. The experimental arrangement is represented in fig. 1, where g represents the heated 1 Proc. Roy. Inst. vol. vi. p. 9. 48 THE FLOATING-MATTER OF THE AIR. lamp-glass, and b the black gap cut out of the beam at its brightest point. The deeper portions of the lungs were thus proved to be filled with optically pure air, which, as such, had no power to generate the organisms proved by Schwann to be essential to the process of putrefaction.! It seemed that this simple method of examination could not fail to be of use to workers in this field. They had hitherto proceeded less by sight than by insight, being in general unable to observe the physical character of the medium in which their experiments were con- ducted. But the method has not been much turned to account ; and this year (1875) I thought it worth while to devote some time myself to the more complete demonstration of its utility. T also wished to free my mind, and if possible the minds of others, from the uncertainty and confusion which now beset the doctrine of ‘spontaneous genera- tion.’ Pasteur has pronounced it ‘a chimera,’ and ex- pressed the undoubting conviction that this being’so it is possible to remove all parasitic diseases from the earth. To the medical profession, therefore, and through them to humanity at large, this question, if the illustrious French philosopher be correct, is one of the last import- ance. But Pasteur’s labours, which have so long been considered models by most of us, have been subjected to 1 «No putrefaction,’ says Cohn, ‘can occur in a nitrogenous sub- stance if it be kept free from the entrance of new Bacteria after those which it may contain have been destroyed. Putrefaction begins as soon as Bacteria, even in the smallest numbers, are acci- dentally or purposely introduced. It progresses in direct proportion to the multiplication of the Bacteria; it is retarded when the Bac- teria (for example, by a low temperature) develop a small amount of vitality, and is brought to an end by all influences which either stop the development of the Bacteria, or kill them, All bacteri- cidal media are therefore antiseptic and disinfecting.’ —Beitrige tur Biologie der Pflanzen, zweites Heft, 1872, p. 203. PUTREFACTION AND INFECTION. 49 rough handling of late. His reasoning has been criti- cised, and experiments counter to his have been adduced in such number and variety, and with such an appear- ance of circumstantial accuracy, as to render the evi- dence against him overwhelming to many minds. This, I have reason to know, has been the effect wrought, not only upon persons untrained in science, but also upon biologists of eminence both in this country and America. The state of medical opinion in England is correctly described in a recent number of the ‘ British Medical Journal,’ where, in answer to the question, ‘In what way is contagium generated and communicated?’ we have the reply that, notwithstanding ‘an almost incalculable amount of patient labour, the actual results obtained, especially as regards the manner of generation of con- tagium, have been most disappointing. Observers are even yet at variance whether these minute particles, whose discovery we have just noticed, and other disease- germs, are always produced from like bodies previously existing, or whether they do not, under certain favour- able conditions, spring into existence de novo. ‘With a view to the possible diminution of the un- certainty thus described, I beg without further preface to submit to the Royal Society, and especially to those who study the etiology of disease, the following descrip- tion of the mode of procedure followed in this inquiry, and of the results to which it has led. § 2. Method of Experiment. A chamber, or case, was constructed, with a glass front, its top, bottom, back, and sides being of wood. At the back is a little door which opens and closes on hinges, while into the slides are inserted two panes of glass, facing each other. The top is perforated in the 50 THE FLOATING-MATTER OF THE AIR. middle by a hole 2 inches in diameter, closed air-tight by a sheet of india-rubber. This sheet is pierced in the middle by a pin, and through the pin-hole is passed the shank of a long pipette ending above in a small funnel. Fic, 2. A circular tin collar, 2 inches in diameter and 13 inch deep, surrounds the pipette, the space between both being packed with cotton-wool moistened by glycerine. Thus the pipette, in moving up and down, is not only firmly clasped by the india-rubber, but it also passes PUTREFACTION AND INFECTION, 51 through a stuffing-box of sticky cotton-wool. The width of the aperture closed by the india-rubber secures the free lateral play of the lower end of the pipette. Into two other smaller apertures in the top of the chamber are inserted, air-tight, the open ends of two narrow tubes, intended to connect the interior space with the atmosphere. The tubes are bent several times up and down, so as to intercept and retain the particles carried by such feeble currents as changes of temperature might cause to set in between the outer and the inner air. » The bottom of the box is pierced with two rows of holes, six in a row, in which are fixed, air-tight, twelve test-tubes, intended to contain the liquid to be exposed to the action of the moteless air. The arrangement is represented in fig. 2, where ww are the side windows through which the searching beam passes from the lamp / across the case c; p is the pi- pette, and a, b, are the bent tubes connecting the inner and outer air. The test-tubes passing through the bot- tom of the case are seen below. On the 10th of Septemher, 1875, this case was closed. The passage of a concentrated beam across it through its two side windows then showed the air within it to be laden with floating matter. On the 13th it was again examined. Before the beam entered, and after it quit- ted the case, its track was vivid in the air, but within the case it vanished. Three days of quiet had sufficed to cause all the floating matter to be deposited on the interior surfaces, where it was retained by a coating of glycerine, with which these surfaces had been purposely varnished. § 3. Deportment of Urine. The pipette being dipped into the tubes, fresh urine was poured into eight of them in succession on the 13th 52 THE FLOATING-MATTER OF THE AIR. of September. Each tube was about half-filled with the liquid. The tubes were then immersed in a bath of brine, raised to ebullition, and permitted to boil for five minutes. Aqueous vapour rose from the liquid into the chamber, where it was for the most part condensed, the uncondensed portion escaping, at a low temperature, through the bent tubes at the top. Before the brine was taken away little stoppers of cotton-wool were inserted in the bent tubes, lest the re-entrance of the air into the cooling-chamber should at first be forcible enough to carry motes along with it. As soon, however, as the outside temperature was assumed by the air within the case the cotton-wool stoppers were removed. The front and back of this chamber were squares of 14 inches the side, the depth of the chamber being 8°5 inches. It contained, therefore, 1666 cubic inches of air, which had unimpeded access to the liquid in the tubes. No stoppers were employed. The air was un- affected by calcination, or even by filtering. Neither cotton-wool nor hermetic sealing was resorted to. Self- subsidence was the only means employed to rid the *untortured ’ air of its floating matter. A second series of eight tubes were filled at the same time with the same liquid, and subjected to the same boiling process. The only difference between the two series was, that these latter tubes were placed in a stand beside the case containing the former ones and exposed to the common air of the laboratory. For the sake of distinction I will call the tubes opening into the case the protected tubes, and those openiug into the common air the exposed tubes. On the 17th of September all the protected tubes were bright and clear, while all the exposed tubes were distinctly turbid. Specks of mould, moreover, were in every case seen on the surface of the exposed liquid. PUTREFACTION AND INFECTION. 53 These waxed daily larger, and finally formed a thick layer on the top of every column. The liquid, mean- while, changed from a pale sherry to a reddish-brown colour. To me the experiment was impressive in the highest degree. On the 27th of September I provided myself with a microscope having a magnifying power of 1200 diame- ters. Under its scrutiny the turbidity of the liquid immediately resolved itself into swarms of Bacteria in active motion. Cohn correctly explains the turbidity. The index of refraction of the Bacterium being slightly different from that of the surrounding medium, a scat- tering of light is the consequence. This scattering, however, and the opalescence it produces, are practically independent of the motions of the Bacteria. Since the date here referred to the exposed liquid has been frequently examined, both with the eye and with the microscope. To the former it is thickly turbid, to the latter it is swarming with life. Its smell is putrid. All this time the protected tubes exhibit a liquid per- fectly unchanged in appearance. For four months it has remained as transparent and of as rich a colour as the brightest Amontillado sherry. On the 1st of October another experiment similar in principle to that just described was begun. Fresh urine was employed, and a much smaller case. The capacity of the latter was 451 cubic inches ; and three test-tubes, instead of twelve, were passed air-tight through its bot- tom. Like those in the larger chamber they were filled by a pipette, and boiled for five minutes in a bath of brine. Beside them were placed three other tubes con- taining the same liquid treated in exactly the same way, but exposed to the common air. On the Sth all the exposed tubes were turbid, and found by microscopic examination to be swarming with Bacteria. The colour 54 THE FLOATING-MATTER OF THE AIR. of the exposed liquid had changed from a pale sherry colour to a brown orange. On the 25th the tubes were again examined, and found full of Bacteria. Two months subsequent to this latter date the infusion, diminished by evaporation, was found well charged with Bacterial life. While this process of putrefaction was goung on outside, the tubes opening into the moteless air of the case remained perfectly clear and void of life. The large chamber represented in fig. 2, and above described, was the first operated on, and the liquid is shown by the draughtsman as filling only a small portion of the test-tubes. This smallness of volume is in part due to evaporation. Test-tubes 1:2 inch wide and 9 inches long were, in all subsequent experiments, nearly filled with the infusions. Strong in the first instance, these were sometimes kept until slow evaporation through the bent tubes at the top of the case had reduced them to one third or one fourth of their original volume. Each experiment, therefore, was, in reality, a series of experi- ments, extending over months, on infusions of different strengths, the concluding ones of the series attaining a very high degree of concentration. § 4. Mutton-Infusion. A new case was constructed to contain six test-tubes. It, like the others, had a front of glass, side windows, anda back door. Its capacity was 857 cubic inches. It was sealed up on the 21st of September, and found free from floating matter on the 24th. The lean of mutton, cut into small pieces, was digested, or soaked, for four hours in water of a temperature of 120° F.!' The infu- ' The temperature recommended by the supporters of sponta- neous generation. PUTREFACTION AND INFECTION, 55 sion was then carefully filtered, and introduced into the six test-tubes by a pipette which was never removed from the case. The mutton-juice was of a fine ruby colour; but on boiling, its albumen was precipitated, subsequently sank, and carried the colouring-matter with it. The superna- tant liquid was perfectly clear. The frothing was con- siderable when the boiling began. Beside this new case was placed a stand containing six test-tubes filled with the same infusion, but exposed to the common air. On the 27th all the outside flasks were perceptibly turbid; on the 28th they were found well filled with Bacteria, which on the 30th had increased to astonishing swarms. On the 15th of October the tubes were again examined, and found charged with undiminished life. They remained thus ‘ putrid’ until the 14th of Novem- ber. During the whole of this time the infusion in con- tact with the moteless air of the chamber remained as clear as distilled water, and entirely free from life. On the 14th of November I infected one of the clear tubes by introducing into it through the pipette a few drops of mutton-infusion which had been prepared and exposed upon the 12th of November, and which two days had sufficed to render turbid. On the 15th the inoculated infusion showed signs of turbidity, and on the 16th putrefaction had actively set in, the liquid heing thickly muddy and full of life. With a moteless chamber and three tubes, experi- ments were subsequently made on a second infusion of mutton. In this case, however, the infusion was boiled, its albumen was precipitated, and removed by filtration prior to its introduction into the chamber. The pellucid liquid was introduced on the Ist of October, boiled for 4 56 THE FLOATING-MATTER OF THE AIR. five minutes in the brine-bath, and abandoned to the air ot the case. A series of exposed tubes containing the same infusion, similarly treated, was placed beside the protected ones. On the 4th all the outside tubes were muddy and swarming with Bacteria. Schroeder and Cohn have shown that different colours are produced by different kinds of Bacteria. In the three exposed tubes here referred to a yellow-green pigment was developed. More than three months after its preparation, the infusion, considerably diminished by evaporation, re- mained in all the protected tubes as clear as at first. § 5. Beef-Infusion. A beef-steak, after having its fat removed, was cut up into small pieces, and digested for three hours at a temperature of 120° F. The liquid was then poured off, boiled, and filtered. It was as clear and colourless as pure water. On the 4th of October it was introduced into three tubes protected by a chamber of 451 cubic inches capacity. It was boiled for five minutes in a brine-bath. Three exposed tubes, containing the same infusion, were placed beside the protected ones. On the 5th the exposed tubes showed signs of haziness, on the 6th they were turbid, of a green colour, and filled with Bacteria. They have maintained for months their mud- diness, colour, and swarming life. While the exposed beef-infusion putrefied in this way, all the protected infusions remained perfectly sweet and clear. § 6. Haddock-Infusion. The haddock was cut up and digested on the 24th of September; it was afterwards introduced into six tubes, protected bya chamber. On boiling, its albumen, PUTREFACTION AND INFECTION. 57 like that of the mutton first referred to, coagulated and sank to the bottom, leaving a perfectly clear liquid overhead. Six exposed tubes filled with the same infu- sion were placed beside the six protected ones. On the 27th the exposed tubes were all turbid and swarming with Bacteria. On the 29th one of the tubes showed a fine green colour; three other tubes showed the same colour afterwards. The vivacity of the organisms was extraordinary, and their shapes various. They darted rapidly to and fro across the field, clashing, recoiling, and pirouetting—rendering it, indeed, difficult to believe in the vegetable nature which the best microscopists assign to them. For nearly three weeks the protected tubes remained perfectly clear. To gain room, the case was subse- quently shifted, and soon afterwards one of the six tubes became turbid with organisms, the germs of which had obviously been shaken into the tube. For more than a month this single infected flask remained in company with the five healthy ones. The air containing the gaseous products of putrefaction had free access to the whole of them, but there was no spread of the infection. As long as the organisms themselves were kept out of the flasks, the ‘sewer-gas’ developed by the putrefaction had no infective power. On the 14th of November I infected two of the five perfectly pellucid tubes with haddock-infusion which, after boil- ing, had been exposed for two days to the air. On the 15th the two tubes had obviously yielded to the infec- tion. On the 16th disease, if I may use the term, had completely taken possession of them. Into one of them only one or two drops of the turbid infusion had fallen, while ten times this amount was introduced into the other. Nevertheless on the 16th both appeared equally turbid. The infection acted exactly like the virus of 58 THE FLOATING-MATTER OF THE AIR. smallpox, a small quantity of which will in the long run produce the same effect as a large one. § 7. Turnip-Infusion. Turnip-juice had a special interest for me in con- sequence of the important part it plays in the experi- ments of heterogenists. I turned to it with the anxious desire to learn whether the statements made concerning it were correct. The conditions laid down as to the strength of the infusion, the temperature to be maintained during the process of digestion, and the time it was to be maintained! were scrupulously adhered to. Thus the turnip was cut into thin slices, and digested for four hours in a beaker of water immersed in a water-bath kept at a tempera- ture close to 120° Fahr. The infusion was then carefully filtered, introduced through a pipette into its case, and boiled there for five minutes. Six protected test-tubes were charged with the infusion on the 24th of Sep- tember, while six other tubes were placed on a stand outside, and exposed to the common aur of the labora- tory. On the 27th the exposed tubes were distinctly turbid, and on microscopic examination were found peopled with Bacteria. The protected tubes, on the contrary, were perfectly clear. A little distilled water had been added to one of the outer tubes. The germi- nal matter, whatever it may be, must have been copious in the water; for the tube to which it was added far exceeded the other two in the rapidity of lite- development. On the 30th this tube contained Bacterict in swarms, of small size, but of astonishing activity. The other tubes also were fairly charged with organisms, ' Bastian, ‘Beginnings of Life,’ vol. i. p. 357, note. PUTREFACTION AND INFECTION. 59 larger and more languid, but not at all so numerous as in the watered tube. On the 5th of October some of the exposed tubes began to clear; as if the Bucteria had died through lack of nutriment, and were falling as a thick sediment to the bottom. During these changes the protected tubes were visibly unaltered, the liquid within every one of them remaining as clear as it had been on the day of its introduction. In this instance I was specially anxious to verify the result by repetition. Two other cases were therefore fitted up to contain three tubes each, and instead of a door a movable panel was placed at the back. After two or three days’ rest both cases were found free from floating matter, and on the Ist of October the turnip- infusion was introduced, and boiled for five minutes in a bath of brine. In the former experiment the temperature of di- gestion was maintained by keeping the beaker contain- ing the turnip in a bath of warm water. In the present instance the turnip was sliced in a dish and placed before a fire. An occult but efficient power like that already ascribed to the actinic rays', might, I thought, be ascribed to radiant heat, and I therefore copied to the letter the mode of digesting pursued by modern heterogenists. Adjacent to the closed cases was placed a series of three exposed tubes, containing a liquid prepared in precisely the same way. On the 4th of October the exposed tubes were all turbid, and swarmed with Bacteria. In two of the tubes they were distinctly more numerous and lively than in the third. Such differences between sensibly conterminous tubes, containing the same infu- sion, are frequent. On the 9th, moreover, the two most 1 «Nature,’ vol. iii. p. 247. 60 THE FLOATING-MATTER OF THE AIR. actively charged tubes were in part crowned by beautiful tufts of Penicillium glaucum.! This expanded gra- dually until it covered the entire surface with a thick tough layer, which must have seriously intercepted the oxygen necessary to the Bacterial life. The Bacteria lost their translatory power, fell to the bottom, and left the liquid between them and the superficial layer clear. _ Another difference, pointing to differences in the unseen life of the air, was shown by these tubes. The turbidity of the two mould-crowned ones was colourless, exhibiting a grey hue. The third tube, the middle one of the three, contained a bright yellow-green pigment, and on its surface no trace of mould was to be seen. It never cleared, but maintained its turbidity and its Bacterial life for months after the other tubes had ceased to show either. It cannot be doubted that the mould- spores had fallen into this tube also, but in the fight for existence the colour-producing Bacteria had the upper hand. Six other tubes, similarly exposed, showed the grey muddiness: all of them became thickly covered with mould, under which the Bacteria died or passed into a quiescent state, fell to the bottom, and left the liquid clear. Up to the 13th of October the purity of the six protected tubes remained unimpaired. Here a complementary experiment was made. It remained to be proved that those long-dormant clear infusions had undergone no change which interfered with their ability to develop and maintain life. On the 13th of October, therefore, the small panel was removed from the back of one of the cases, and with three new pipettes specimens were taken from the three tubes within it. The closest search revealed no living thing. 1} Ordinary mould. PUTREFACIION AND INFECTION. 61 The air of the laboratory being permitted to diffuse freely into the case, on the day after the removal of the panel the test-beam showed the case to be charged with floating: matter. The access of this matter was the only condition necessary to the production of life; for on the 17th all the tubes were muddy and swarming with Bacteria. A similar experiment, subsequently made, revealed some of the snares and pitfalls which await an in- cautious worker. The chamber already referred to as containing six tubes, filled with turnip-juice, pre- served the infusion clear for a month. On the 21st of October the back door of the chamber was opened, and specimens of the clear infusion were taken out for examination by the microscope. The first tube examined showed no signs of life. This result was expected. Picking up another pipette, I took a sample from the second tube. Here, to my astonish- ment, the exhibition of life was monstrously copious. There were numerous globular organisms, which re- volved, rotated, and quivered in the most extraor- dinary manner. There were also numbers of lively Bacteria darting to and fro. An experimenter who ponders his work and reaches his conclusions slowly, cannot immediately relinquish them: and in the pre- sent instance some time was required to convince me that no mistake had been made. I could find none, and was prepared to accept the conclusion that in the boiled infusion, despite its clearness, life had appeared. But why, in the protected turnip-infusion, which had been examined on the 13th of October, could no trace of life befound? In this case perfect transparency was accompanied by an utter absence of life. The selfsame action upon light that enabled the Bacteria to show themselves in the microscope must, one would 62 THE FLOATING-MATTER OF THE AIR. think, infallibly produce turbidity. Why, moreover, should life be absent from the first member of the present group of tubes? I searched this again, and found in it scanty but certain signs of life. This augmented my perplexity. A third tube also showed traces of life. I reverted to the second tube, where life had been so copious, and found that in it the organisms had become as scanty asin the others. I confined myself for a time to the three tubes of the first row of the six, going over them again and again; sometimes finding a Bacterium here and there, but sometimes finding nothing. The first extraordinary exhibition of life it was found impossible to restore. Doubtful of my skill as a microscopist, I took specimens from the three tubes and sent them to Prof. Huxley, with a request that he would be good enough to examine them. On the 22nd the search was extended to the whole of the tubes. Early in the day lively Bacteria were found in one of them; later on, not one of the six yielded to my closest scrutiny any trace of life. On the evening of the 22nd a note was received from Prof. Huxley stating that a careful examination of the speci- mens sent to him revealed no living thing. Pipettes had been employed to remove the infusion from the test-tubes. They were short pieces of narrow glass tubing, drawn out to a point, with a few inches of india-rubber tubing attached to them. This was found convenient for bending so as to reach the bottom of the test-tubes. Suspicion fell upon this india-rubber. It was washed, the washing-water was examined, but no life was found. Distilled water had been used to cleanse the pipettes, and on the morning of the 23rd I entered the laboratory intending to examine it. Before dip- ping a pipette into the water I inspected its point. PUTREFACTION AND INFECTION. 63 The tiniest drop had remained in it by capillary attrac- tion from the preceding day. This was blown on toa slide, covered, and placed under the microscope. An astonishing exhibition of life was my reward. Thus on the scent, I looked through my pipettes, and found two more with the smallest residual drops at the ends: both of them yielded a field rampant with life. The Bacteria darted in straight lines to and fro, bending right and left along the line of motion, wriggling, rotating longi- tudinally, and spinning round a vertical transverse axis. Monads also galloped and quivered through the field. From one of these tiny specks of liquid was obtained an exhibition of life not to be distinguished from that which had astonished me on the 21st. Obviously the phenomenon then observed was due to the employment of an unclean pipette. Equally obvious is it that in inquiries of this nature the experi- menter is beset with danger, the grossest errors being possible when there is the least lack of care. The chamber here operated on had been opened with a view to testing the capacity of the infusions within it to develop and maintain life. For four weeks they had remained perfectly clear. Two days after the door was opened and the common laboratory air admitted all six tubes were turbid, and swarming with Bacteria. Some of them were very long, and their wrig- gling and darting hither and thither very impressive. The same chamber was again thoroughly cleaned, sealed, and permitted to remain quiet until the floating matter had subsided. On the 17th of November a fresh infusion of turnip was introduced into it through the pipette, boiled in an oil-bath, and again abandoned to the air of the case. After several months the infusion in every tube of the siz remained as clear as it was on the day of its introduction. 64 THE FLOATING-MATTER OF THE AIR. Six other tubes charged with the same infusion, boiled in the same way, became turbid in a few days, and subsequently covered with thick layers of Penicii- lium. § 8. Hay-Infusion. This infusion has been credited with a power of spontaneous generation similar to that ascribed to turnip-juice. The hay being chopped into short lengths was digested for four hours in water kept at a tempera- ture of 120° Fahr. On the 24th of September the filtered infusion was introduced into its chamber, and boiled there for five minutes. Six tubes were charged with the protected liquid, while six other tubes, filled with the same infusion, were placed on a stand outside the case. On the 27th the inside flasks were clear, the outside ones faintly turbid. On the 28th spots of mould ap- peared upon all the exposed surfaces. The infusion in one of the outside tubes had been diluted with distilled water, and in it the development of life was far more rapid than in the five others; all of them, however, on the 28th contained Bacteria. On the 29th I noticed a larger organism than the Bacteria moving rapidly to and fro across the field, the drop containing it being taken from the dilute infusion. Several of these monads weré seen upon the 30th gambol- ling among the smaller Bacteria, appearing bright or” dark as they sank or rose in the liquid, a film of which, large as they looked, was to them an ocean. Swarms of Bacteria were seen on the 2nd of October, their transla- tory motions being so rapid and varied, and so apparently guided by a purpose, as to render it difficult to believe that they could be anything else than animals. On the 15th there was a marvellous exhibition of the larger PUTREFACTION AND INFECTION. 65 Infusoria, which appeared to have driven the Bucteria from their habitat, as few of the latter were to be seen. My inability to find the larger creatures a second time in such numbers perplexed me, causing me to conclude that I had accidentally alighted upon a colony of them- My experience with the unclean pipettes already de- scribed, pointed, however, to another source. While three days sufficed to break down their purity, and to fill the six exposed tubes with Bacterial life, the six protected ones remained for more than three months as clear and healthy as they were on the day the infu- sion was poured into them. Neither a trace of mould upon the surface of any one of them, nor a trace of tur- bidity in its mass, was to be seen. Into another chamber containing three test-tubes a very strong infusion of hay was introduced on the Ist of October. It was boiled for five minutes, and then aban- doned to the air of the case. Three other tubes exposed to the laboratory air were placed on a stand beside the case. The colour of the infusion was very deep, but it was quite transparent. One of the outer tubes was diluted with distilled water. On the 3rd the infusion in this tube was turbid, the other exposed ones remain- ing clear. The unseen germinal matter had in some way or other invaded the distilled water, and made it infective. The dilute infusion contained multitudes of Bacteria, many motionless, but many moving rapidly about. On the 4th of October all the tubes swarmed with Bacteria. They continued muddy till the middle of November, when they were employed for experi- ments on infection. Throughout the whole of this time the protected tubes remained unchanged. With regard to infection, it may be stated here that 66 THE FLOATING-MATTER OF THE AIR. the merest speck of a vegetable infusion containing Bacteria infects all animal infusions, and vice versa. The bursting of a bubble infects an infusion reached by the spray. It is the envelope, and not the gas of the bubble, which produces this result. Other experiments on hay-infusions, acid, neutral, and alkaline, placed in contact with air purified in various ways, yielded in 1875 the same negative re- sult. § 9. Infusion of Sole. The fish was cut up and digested for three hours in water kept at 120° Fahr. On the 17th of November it was introduced into a case containing three test-tubes, and boiled there for five minutes. Three other tubes hung outside the case were exposed to the ordinary laboratory air. The three exposed tubes were feebly but distinctly cloudy on the 19th. On the 22nd they were all thickly turbid. Scattered spots of Penicillium then appeared on two of them, while the third tube, which stood be- tween these two, kept the Penicillium down. This central tube contained the pigment-forming Bacteria, which have frequently shown a singular power in pre- venting the development of mould. For nearly two months the central tube successfully withstood this de- velopment, while its two neighbours were covered by a matted layer of Penicillium. During the whole of this time the protected infusion continued as clear and colourless as distilled water. § 10. Liver-Infusion. On the 10th of November the infusion was prepared by the process of digesting already so often described. PUTREFACTION AND INFECTION. 67 It was introduced into a case containing three protected tubes, and boiled there for five minutes in the brine- bath. Hung on to the chamber at the same time were three tubes containing the same infusion, but exposed to the common air. On the 13th Bacteria were numerous in the exposed tubes, and soon afterwards all three of them became thickly muddy and putrescent. They continued so for months. The protected tubes, on the contrary, showed throughout a bright yellow liquid, as transparent and fresh as it was on the day of its introduction into the case. § 11. Infusions of Hare, Rabbit, Pheasant, and Grouse. For the sake of economy, as so many of them were employed, the shape of the cases was subsequently varied. The rounded end of a tall glass shade was cut off, so as to convert the shade into a hollow cylinder, open at both ends. This was set upright on a wooden stand, and cemented to it air-tight. Through the stand passed three large test-tubes also air-tight: To the top of the cylinder was cemented a circular piece of wood, the middle of which was occupied by a pipette passing first through india-rubber and then through a stuffing-box of cotton-wool moistened by glycerine.' The air within the case was connected with the air without by means of the open bent tubes already described. In the first experiments made with these cases defects of construction were revealed during the boiling of the infusions. But increased experience enabled me to 1 In the earlier experiments the india-rubber formed the bottom of the stuffing-box, where particles were sometimes detached from it by the motion of the pipette. To prevent this the positions of wool and rubber were afterwards reversed. 68 THE FLOATING-MATTER OF THE AIR. render them secure. The floating matter within the cases having been permitted to subside, into four of them, on the 30th of November, infusions of hare, rabbit, pheasant, and grouse were introduced. The infusions were boiled in the usual way, and abandoned to the air of the case. Outside each case, and hung on to it, were three test-tubes of the same size and containing the same infusion as that within. Examined on Christmas-day, the following were the observed results :— Pheasant.—The three interior tubes perfectly limpid : the three exposed ones turbid and covered with Pent- cillium. Grouse.—In the same condition as the pheasant. Hare.—The same as grouse and pheasant. Rabbit.—The three interior tubes covered with tufts of particularly beautiful Penicilliwm, some of the tufts striking deep into the liquid. In two out of the three tubes, moreover, mycelium was flourishing below. All the outer tubes were, as usual, turbid and covered with Penicilliwm. Is this, then,.a case of spontaneous generation ? Without further evidence no cautious worker would draw such a conclusion. Opposed to this isolated in- stance stand all the others mentioned in these pages, and their proper action on the mind is to compel it to demand the closest scrutiny before accepting this apparent exception as a real one. Subjected to such scrutiny, it appeared that of the four shades the one containing the rabbit-infusion, and that only, had yielded to the heat of boiling. The shade had been fastened upon its slab with plaster and cement, which became so loose during the boiling that the steam issued from the chinks. But crannies which could per- mit steam to escape could permit air to enter, and to PUTREFACTION AND INFECTION. 69 the presence of such air the appearance of the Peni- cilliwm was doubtless due. I did not, however, rest content with mere inference, but tested the rabbit-infusion by placing three fresh tubes of it in one of the firmer cases first described. It was introduced and boiled on the 5th of January, 1876, three other tubes filled with the same boiled infusion being exposed on the same day to the ordinary air. The three protected tubes remained clear for three months, while in three days the three exposed ones were charged with Bacteria. Salmon.—The colouring-matter of this fish did not at all affect the infusion ; indeed, no better example of original freedom from colour or opalescence, and of persistent purity in contact with the moteless air, has occurred to me than salmon-infusion. It was introduced into a cylindrical case on the 13th of December, where it continued for months to show the brilliant trans- parency exhibited at first. Three unprotected tubes, on the other hand, became turbid and covered with mould in a few days. Hops.—One tube of this infusion was protected simply by a lamp-glass, corked and cemented above and below. Through the lower cork passed the single test- tube, air-tight ; while through the upper one passed the pipette and the bent tubes intended to connect the outer and the inner air. The infusion was prepared and in- troduced on the 28th of October. In a few days the exposed tube was found turbid and covered with mould; the protected tube, on the contrary, remained clear for several months. Tea and Coffee.—One tube of each was protected by a lamp-glass similar to that employed in the infusion of hops. Both were prepared on the 28th of October, exposed tubes being hung up at the same time. The 70 THE FLOATING-MATTER OF THE AIR. protected tea has remained clear, while the exposed tea is turbid and covered with mould. Both the exposed coffee and the protected coffee are, on the other hand, turbid and covered with Penicillium. The remarks already made with regard to the rabbit- infusion apply here. The case is one, not for the hasty admission of spontaneous generation, but for further scrutiny. JI examined the apparatus as it stood. The pipette used to introduce the coffee (and this one only of the three employed in these experiments) rested against the outer edge of the tube containing the infu- sion. This had in part evaporated, had been in part re-condensed, and had trickled down the pipette so as to form a small drop at the point where pipette and tube touched each other. The drop had virtually washed the outer surface of the pipette, carrying with it, in part, such matter as might have attached itself to that sur- face. A portion of this washing-water reaching the infusion was clearly the origin of the life observed. The sure test, however, was the repetition of the experi- ment under conditions which should exclude this source of error. On the 27th of December accordingly two tubes protected by lamp-glasses were prepared, two other tubes of the infusion being exposed to the air. The former remained clear for months, the latter in the same number of days became turbid and covered with Penicillium. § 12. Infusions of Codfish, Turbot, Herring, and Mullet. With a view of causing these experiments on mote- less and mote-laden air to run parallel with others made with hermetically-sealed tubes, to be described further on, I added the fish named in the heading of this section to the other substances examined. The mullet was in- PUTREFACTION AND INFECTION. 71 troduced into its case on the 3rd of January. The warm air of the room had, however, so acted on the wood of the chamber, which had been employed in former ex- periments, that the water of condensation trickled from a chink in the bottom. The other chambers were mended as far as possible, and into them the infusions were introduced on the 4th of January. Each chamber, as before, was provided with three exposed tubes for comparison with three protected tubes within. On the morning of the 6th the exposed turbot-infusion was clear in all the tubes ; a few hours subsequently two out of the three became cloudy; while on the 7th Bacteria had taken. possession of all of them. All the unpro- tected tubes of codfish were cloudy on the 6th, more cloudy on the 7th, and covered with a soapy layer upon the 8th. The three exposed herring-tubes were also cloudy on the 6th, the cloudiness advancing afterwards to thicker turbidity. The mullet gave way in the same manner. For more than three months the protected tubes, including even the wmperfect chamber which protected the mullet-infusion, have remained as clear as they were upon the day of their introduction. To these fish-infusions may be added others of eel and oyster. Two tubes of each, protected by lamp- glasses, were charged on the 27th of December. They remain unchanged. Two other pairs of tubes, prepared in the same way and exposed to the laboratory air, are turbid and covered with Penicillium. § 13. Infusions of Fowl and Kidney. Three tubes.of the fowl-infusion were introduced into a case, and boiled there for five minutes, on the 4th of January. Three similar tubes were at the same time exposed to the air. On the 6th all the outer 72 THE FLOATING-MATTER OF THE AIR. tubes were cloudy, the cloudiness becoming denser on the following days, while disks of Penicillium began to form on the exposed surfaces. It was found ex- ceedingly difficult to obtain a clear infusion of kidney. The liquid, after it had passed through a dozen filters, was still quite muddy. With considerable labour and care, and by the employment of 200 filters, the mechani- cally suspended matter was at length removed, and a clear infusion obtained. It was introduced into its case, to which three exposed tubes were attached, on the 4th of January. On the 7th the latter were per- ceptibly cloudy, on the 8th distinctly so, while specks of mould rested upon them all. The protected tubes, on the contrary, have for months maintained their trans- parency undimmed.! The entire number of experiments made to illustrate the association of scattered light and Bacterial and fungoid life are not here recounted. Whiting, for example, may be added to the fish, and pork to the flesh examined, while many of the other substances have been tested oftener than I have thought it neces- sary to record. The method of boiling was also varied in a manner which may claim a passing reference here. § 14. Boiling by an Internal Source of Heat. Two large test-tubes were fixed air-tight in the same case. On the 8thof November, after the floating matter had subsided, infusions of hay and turnip were introduced. Dipping deep into each infusion were two tinned copper wires, connected below by a spiral of 1 Kidney has been mentioned by Dr. Bastian as a substance with which he demonstrates the occurrence of spontaneous genera- tion. He does not mention the extraordinary turbidity of the infusion, which proved so troublesome to me. PUTREFACTION AND INFECTION. 73 platinum wire. The copper wires passed through the case, and were connected with a voltaic battery outside. The spiral was heated by the current from this bat- tery. After a few minutes’ heating ebullition set in, and was continued for five minutes in each tube. Two other tubes containing the same infusions were boiled in the same way, and afterwards hung outside the case containing the two protected tubes. In another chamber were placed two tubes con- taining infusions of beef and mutton. The arrange- ment and the treatment were precisely the same as those just described in the case of hay and turnip. Examined some months subsequently, the exposed tubes of all four infusions were found turbid and covered with Penicillium, while all the four protected tubes remained unchanged. During the boiling process some flocculi detached themselves from the tinned sur- faces of the copper wires; but in the protected tubes these have fallen to the bottom, and left the supernatant liquid clear. Platinum wires would have been better than tinned copper ones. § 15. Partial Discussion of the Results. Thus by experiments, reiterated in many cases, with urine, mutton, beef, pork, hay, turnip, tea, coffee, hops, haddock, sole, salmon, codfish, turbot, mullet, herring, eel, oyster, whiting, liver, kidney, hare, rabbit, fowl, pheasant, grouse, has the induction been established, that the power of developing Bacterial life by atmospheric air and its power of scattering light, go hand in hand. We shall immediately examine more closely what this means. The necessity of employing strong infusions has been frequently dwelt upon, if we would realize the. 74 THE FLOATING-MATTER OF THE AIR. phenomena of spontaneous generation. I would there- fore recall to mind what has been stated on a previous page, that in most of the experiments here described the infusion at starting was strong, and that it was permitted to evaporate with extreme slowness until its concentration became three or four fold what it had been at starting. Every experiment was thus converted into an indefinite number of experiments on infusions of different strengths. Never, in my opinion, was the requirement as to concentration more completely ful- filled, and never was the reply of Nature to experiment more definite and satisfactory. The temperatures, moreover, to which the infusions have been sub- jected embrace those hitherto alleged to be effectual, extending indeed beyond them in both directions.! They reached from a lower limit of 50° to a higher limit of more than 100° Fahr. Still higher tempera- tures were applied in other experiments to be described subsequently. With regard to the number of the infusions, more than fifty moteless chambers, each with its system of tubes, have been tested. There is no shade of uncertainty in any of the results. In every instance we have, within the chamber, perfect limpidity and sweetness—without the chamber, putridity and its characteristic smells. Inno instance is the least counte- nance lent to the notion that an infusion deprived by heat of its inherent life, and placed in contact with air cleansed of its visibly suspended matter, has any power whatever to generate life anew. If it should be asked how I have assured myself that. the protected liquids do not contain Bacteria, I would, in the first place, reply that with the most careful microscopic search I have been unable to find 1 See Proc. of Roy. Soc. vol. xxi. p, 180, where a temperature of 70° is described as effectual. PUTREFACTION AND INFECTION, 75 them. But much more than this may be affirmed. The electric or the solar beam is a far more powerful and searching test in this matter than the microscope. In the foregoing pages I have more than once described the clearness of my protected infusions, after months of exposure, as equal to that of distilled water. So far is this from being an exaggeration, that it falls short of the truth; for I have never seen distilled water so free from suspended particles as the protected infusions prove themselves tobe. When for months a transparent liquid thus defies the scrutiny of the searching beam, maintaining itself free from every speck which could scatter light asa Bacterium scatters it—when, moreover, an adjacent infusion, prepared in precisely the same way, but exposed to the ordinary air, becomes first hazy, then turbid, and ends by wholly shattering the concentrated beam into irregularly scattered light, I think we are entitled to conclude that Bacteria are as certainly absent from the one as they are present in the other. (See Note I. at the end of this paper.) For the right interpretation of scientific evidence something more than mere sharpness of observation is requisite, very keen sight being perfectly compatible with very weak insight. I was therefore careful to have my infusions inspected by biologists, not only trained in the niceties of the microscope, but versed in all the processes of scientific reasoning. Their conclusion is that it would simply weaken the demonstrative force of the experiments to appeal to the microscope at all. § 16. Suspended Particles in Air and Water ; their relation to Bacteria. Examined by the concentrated solar rays, or by the condensed electric beam, the floating matter of the air 76 THE FLOATING-MATTER OF THE AIR. is seen to consist :—first, of particles so coarse that their individual motions can be followed by the eye ; secondly, of a finer matter which is not to be distinguished as motes, but which emits a uniform and changeless light. In this finer matter the coarser motes move as in a medium. ; As regards the production of colour, the action of small particles has been examined by Briicke in a paper ‘On the Colours of Turbid Media.’! In relation to the question of polarization, Professor Stokes has made some remarks in his memoir ‘ On the Change of the Refrangi- bility of Light.’? I may also be permitted to refer to my own papers ‘On New Chemical Reactions by Light’ and ‘On the Blue Colour of the Sky,’ in the Proceed- ings of the Royal Society for 1868-69, and to a paper ‘On the Action of Rays of High Refrangibility on Gaseous Matter,’ in the Philosophical Transactions for 1870. M. Soret, Lord Rayleigh, and Mr. Bosanquet have also worked at this subject, which, as far as it now concerns us, a few words will render clear. When the track of a parallel beam in dusty air is looked at horizontally through a Nicol’s prism, in a direction perpendicular to the beam, the longer diago- nal of the prism being vertical, a portion of the light from the finer matter, being polarized, is extinguished. The coarser motes, on the other hand, which do not polarize the light, flash out with greater force, because of the increased darkness of the space around them. The individual particles of the finest floating matter of the air lie probably far beyond the reach of the microscope. At all events it is experimentally demon- strable that there are particles which act similarly upon light, and which are entirely ultra-microscopic. A few * Pogg. Ann. lxxxviii. p. 363. 2 Philosophical Transactions, vol. 142, pp. 529-530. PUTREFACTION AND INFECTION, 77 days ago, for example, an inverted bell-jar was filled with distilled water, into which, while it was briskly beaten by a glass rod, was dropped a solution of mastic in alcohol. The proportion was less than that employed by Briicke, being about ten grains of the gum to 1,000 grains of the alcohol. The jar was placed under a sky- light, at the height of the eye above the floor. It was of a beautiful cerulean hue, this colour arising wholly from the light scattered by the mastic particles. Looked at horizontally through a Nicol’s prism, with its shorter diagonal vertical, the blue light passed freely to the eye. Turning the long diagonal vertical, the scattered light was wholly quenched, and the jar appeared as if filled with ordinary pure water. I tried the effect of a powerful filter upon those parti- cles, and found that they passed sensibly unimpeded through forty layers of the best filtering-paper.! The liquid containing them was examined by a microscope magnifying 1,200 diameters. The suspended mastic particles entirely eluded this power, the medium in which they swam being as uniform as distilled water in which no mastic whatever had been precipitated. The optical deportment of the floating matter of the air proves it to be composed, in part, of particles of this excessively minute character. The concentrated beam reveals them collectively, long after the micro- scope has ceased to distinguish them individually. In London rooms, moreover, they are for the most part organic particles, which may be removed from the air by combustion. In presence of such facts, any argu- ment against atmospheric germs, based upon their being beyond the reach of the microscope, loses all validity. We are here brought face to face with a question 1 There are filters, however, which stop them; but of this immediately. 78 THE FLOATING-MATTER OF THE AIR. of extreme importance, which it will be useful to clear up. ‘Potential germs’ and ‘ hypothetical germs’ have been spoken of with scorn, because the evidence of the microscope as to their existence was not forthcoming. Sagacious writers had drawn from their experiments the perfectly legitimate inference that in many cases the germs exist, though the microscope fails to reveal them. Such inferences, however, have been treated as the pure work of the imagination, resting, it was alleged, on no real basis of fact. But in the concentrated beam we possess what is virtually a new instrument, exceeding the microscope indefinitely in power. Directing it upon media which refuse to give the coarser instrument any information as to what they hold in suspension, these media declare themselves to be crowded with particles —not hypothetical, not potential, but actual and myriad- fold in number—showing the microscopist that there is a world far beyond his range. In §§ 6 and 8 experiments on the infection of clear infusions by others containing visible Bacteria are re- ferred to. But for the infection to be sure it is not necessary that the Bacteria should be visible. Over and over again I have repeated the experiments of Dr. Bur- don Sanderson on the effective power of ordinary distilled water, in which the microscope fails to reveal a Bacte- rium. The water, for example, furnished to the Royal- Institution laboratory by Messrs. Hopkin and Williams is sensibly as infectious as an infusion swarming with Bacteria. The vessels are the source of infection here. Perhaps the severest experiment of this kind ever made was one executed by Dr. Sanderson with water prepared by myself. In 1871 I sought anxiously and assiduously for water free from suspended particles. The liquid was obtained in various degrees of purity, but never entirely pure. Knowing the wonderful power of PUTREFACTION AND INFECTION. 79 extrusion, as regards foreign matter, brought into play by water in crystallizing, the thought occurred to me of examining the liquid derived from the fusion of the most transparent ice. At my request, therefore, my assistant arranged the following apparatus for me:— Fia. 3. Through the plate of an air-pump (fig. 3) passed air- tight the shank of a large funnel. A small glass bulb, B, furnished with a glass stopcock, was attached to the shank of the funnel below. Prior to being put together all parts of the apparatus had been scrupulously cleansed. 5 : 80 THE FLOATING-MATTER OF THE AIR. In the funnel was placed a block of ice, 1, selected for its transparency, having a volume of 1000 cubic inches or thereabouts, and over the ice was placed an air-tigh, receiver. Several times in succession the air was re- moved from this receiver, its place on each occasion being takeu by other air carefully filtered through cotton-wool. The transparent ice was thus surrounded by moteless air. The ice was now permitted to melt; its water trickled into the small glass bulb below, which was filled and emptied a great number of times. From the very heart of the block of ice the water was finally taken and sub- jected to the scrutiny of the concentrated beam. It proved to be the purest liquid I had ever seen—pro- bably the purest human eye had ever seen; but still it contained myriads of ultra-microscopic particles. The track of the beam through it was of the most delicate blue, the blue light being perfectly polarized. It could be wholly quenched by a Nicol’s prism, the beam then passing through the liquid as through a vacuum. A comparison of the light with that scattered by mastic particles such as those above referred to, proved the suspended particles of the ice-water to be far smaller than those of the mastic. No microscope, therefore, could come near them,! Such water, however, was proved by Dr. Sanderson to be as infectious as the water from any ordinary tap. Infinitesimal as these particles are, however, they may be separated by mechanical means from the liquid in which they are held in suspension. Filters of porous earthenware, such as the porous cells of Bunsen’s battery, 1 [ have endeavoured to convey some notion of the smallness of these scattering particles in ‘Fragments of Science,’ Art. ‘Scientific Use of the Imagination.’ See note on Mr. Dallinger’s observations at the end of this Memoir. PUTREFACTION AND INFECTION 81 have been turned to important account in the researches of Dr. Zahn, Professor Klebs, and Dr. Burdon Sander- son. In various instances it has been proved that, as regards the infection of living animals, the porous earthenware intercepts contagia. For the living animal, organic infusions, or Pasteur’s solution, may be substi- tuted. Not only are ice-water, distilled water, and tap-water thus deprived of their powers of infection, but, by plunging the porous cell into an infusion swarm- ing with Bacterial life, exhausting the cell, and per- mitting the liquid to be slowly driven through it by atmospheric pressure, the filtrate is not only deprived of its Bacteria, but also of those ultra-microscopic germs which appear to be as potent for infection as the Bac- teria themselves. The precipitated mastic particles before described, which pass unimpeded through an in- definite number of paper filters, are wholly intercepted by the porous cell. These germinal particles abound in every pool, stream, and river. All parts of the moist earth are crowded with them. Every wetted surface which has been dried by the sun or air contains upon it the parti- cles which the unevaporated liquid held in suspension. From such surfaces they are detached and wafted away, their universal prevalence in the atmosphere being thus accounted for. Doubtless they sometimes attach themselves to coarser particles, organic and inorganic, which are left behind along with them ; but they need no such rafts to carry them through the air, being them- selves endowed with a power of flotation commensurate with their extreme smallness and the specific lightness of the matter of which they are composed. I by no means affirm that the developed Bacterium, which requires for its maintenance nutriment beyond that which ordinary water can always supply, is never 82 THE FLOATING-MATTER OF THE AIR. wafted through the air. Cases may arise favourable to the growth and dispersion of the full-grown or- ganism. Whether, after desiccation, it retains the power of reproduction is another question. But it ought, I think, to be steadily borne in mind that the complete Bacteria and the atmospheric matter from which they spring are, in general, different things. I have care- fully sought for atmospheric Bacteria, but have never found them. They have never, to my knowledge, been found by others; and that they arise from matter which has not yet assumed the Bacterial form is, as just shown, capable of demonstration. An organic infusion, boiled and shielded from atmospheric particles, will remain clear for an indefinite period, while a fragment of glass which has been exposed to the air, but on which no trace of a Bacterium is to be found, will in two or three days develop in the infusion a multitudinous crop of life. We have now to look a little more closely at these particles, foreign to the atmosphere but floating in it, and proved beyond doubt to be the origin of all the Bac- terial life which our experiments have thus far revealed. We must also look at them as they exist in water, in countless multitudes, being as foreign to this medium as the floating atmospheric dust is to the air in which it swims. The existence of the particles is quite as certain as if they could be felt between the fingers, or seen by the naked eye. Supposing them to augment in magni- tude until they come, not only within range of the microscope, but within range of the unaided senses. Let it be assumed that our knowledge of them under these circumstances remains as defective as it is now— that we do not know whether they are germs, particles of dead organic dust, or particles of mineral matter. Suppose a vessel (say a flower-pot) to be at hand filled with nutritious earth, with which we mix our unknown PUTREFACTION AND INFECTION. 83 particles, and that in forty-eight hours subsequently buds and blades of well-defined cresses and grasses appear above the soil. Suppose the experiment when repeated a hundred times to yield the same unvarying result. What would be our conclusion? Should we regard those living plants as the product either of dead dust, or of mineral particles? or should we regard them as the offspring of living seeds? The reply is unavoidable. We should undoubtedly consider the experiment with the flower-pot as clearing up our pre-existing ignorance ; we should regard the fact of their producing cresses and grasses as proof positive that the particles sown in the earth of the pot were the seeds of the plants which have grown from them. It would be simply monstrous to conclude that they had been ‘spontaneously generated.’ This reasoning applies word for word to the develop- ment of Bacteria from that floating matter which the electric beam reveals in the air, and in the absence of which no Bacterial life has been generated. I cannot see a flaw in the reasoning; and it is so simple as to render it unlikely that the notion of Bacterial life being developed from dead dust can ever gain currency among the members of the medical profession. It has been said of those whom the evidence adduced in favour of spontaneous generation fails to convince, that they seem willing to believe in almost any infringe- ment of natural uniformity rather than admit the doctrine.' This surely is an inversion of the true order of the facts. Natural uniformity is the record of expe- rience; and, apart from the phenomena to be accounted for, there is not a vestige of experience, possessed either by the man of science or the human race, which warrants the notion that dead dust, and not living seed, is the source of the crops which spring from our infusions 1 Transactions of the Pathological Society, vol. xxvi. p. 273. 84 THE FLOATING-MATTER OF THE AIR. when impregnated by the floating particles of the at- mosphere. § 17. Recent Experiments on Heterogenesis. The uniform sterility of the boiled infusions described in the foregoing pages, when protected from the floating matter of the air, proves that they do not contain germs capable of generating life. Our most advanced hetero- genist, indeed, affirms that a temperature of 140° Fahr. reduces, in all cases, such germs to a state of actual or potential death; and he ingeniously argues that as, even in flasks which have been raised to a temperature of 212°, and hermetically sealed, putrefaction, and its associated Bacteria, do most certainly arise, such Bac- teria must be spontaneously generated. ‘We know,’ he says, ‘ that boiled turnip or hay-infusions, exposed to ordinary air, exposed to filtered air, to calcined air, or shut off altogether from contact with air, are more or less prone to swarm with Bacteria and Vibriones in the course of from two to six days.’! We are here met by a difficulty at’ the outset. The proof of Bacterial death at 140° Fahr. consists solely in the observed fact, that when a certain liquid is heated to that temperature no life appears in it afterwards ; while in another liquid life appears two days after it has been heated to 212°. Instead of concluding that in the one liquid life is destroyed and in the other not, it is assumed that 140° Fahr. is the death-temperature for both ; and this being so, the life observed in the second liquid is regarded as a case of spontaneous generation. A great deal of Dr. Bastian’s most cogent reasoning rests upon this foundation. Assumptions of this kind guide him in his most serious experiments. He finds, } «Evolution and the Origin of Life.’ p. 94. PUTREFACTION AND INFECTION. 85 for example, that a mineral solution does not develop Bacteria when exposed to the air; and he concludes from this that an organic infusion also may be thus ex- posed without danger of infection. He exposes turnip- juice accordingly, obtains a crop of Bacteria, which, in the light of his assumption, are spontaneously generated. Such are the warp and woof of some of the weightiest arguments on this question which have been addressed by him to the Royal Society.! Granting, then, all that Dr. Bastian alleges regard- ing his experiments to be correct, the logical inference would be very different from his inference. In a future essay his position will be more clearly defined. To the examination of his experiments I now address myself. § 18. Experiments with Filtered Air. A bell-jar containing about 700 cubic inches of air was firmly cemented to a slab of wood coated with resin, and supported on three legs.2_ Through the slab passed, air-tight, three large test-tubes (4, 3, ¢, fig. 4). Prior to cementing, the tubes had been three-fourths filled, one with an infusion of hay, another with an infusion of turnip, and a third with an infusion of mutton. On the 2nd of November the mote-laden air was pumped out, air slowly filtered through a long tight plug of cotton-wool being allowed to take its place. The jar was emptied and refilled until the closest scrutiny by a concentrated beam revealed no floating matter within it. The infusions were then boiled for five minutes, } Proceedings, vol. xxi. p. 130. 2 Two hoops of sheet iron, with an annular space about an inch wide, were fastened on to the slab of wood. The annular space was filled with hot cement, into which the hot bell-jar was pressed. The circular space within the smaller hoop was also covered by a layer of cement, 86 THE FLOATING-MATTER OF THE AIR. and abandoned to the air of the jar. During ebullition a small quantity of the liquid in one of the tubes boiled over, and rested upon the interior resinous surface at a little distance from the mouths of two of the tubes. The germinal matter, it may be remarked, is not readily blown away from such a surface, and it certainly was not wholly removed by our feeble current of filtered air. Three exposed tubes containing the same infusion were placed at the same time beside the protected ones Fie. 4. In three days these exposed tubes became turbid and charged with life ; but for three weeks the infusions in contact with the filtered air remained perfectly clear. At the end of three weeks, that is on the 23rd of November, I desired my assistant to renew the air in the bell-jar. He pumped it out, and while permitting fresh air to enter through the cotton-wool filter, my attention PUTREFACTION AND INFECTION. 87 was directed toa couple of small round patches of Peni- cilliwm resting in the liquid that had boiled over on the resin. I at once made the remark that the experiment was a dangerous one, as the entering air would probably detach some of the spores of the Penicilliwm and diffuse them in the bell-jar. This was, therefore, filled very slowly, so as to render the disturbance a minimum. Next day, however, a tuft of mycelium was observed at the bottom of one of the three tubes, namely that con- taining the hay-infusion. It soon grew large enough to fill a considerable portion of the tube. For nearly a month longer the two tubes containing the turnip- and mutton-infusions maintained their transparency unim- paired. Late in December the mutton-infusion, which was in dangerous proximity to the outer mould, showed a tuft of Penicilliwm upon its surface. The beef- infusion continued bright and clear for nearly a fort- night longer. The cold winter weather caused me to add a third gas-stove to the two which had previously warmed the room where the experiments are conducted. The warmth of this stove played upon one side of the bell-jar ; and on the day after the lighting of the stove, the beef-infusion gave birth to a tuft of mycelium. In this case the small spots of Penicillium might have readily escaped attention; and had they done so we should have had here three cases of ‘spontaneous gen- eration’ more striking than most of those that have been adduced in support of this doctrine. The experiment was subsequently made upon a larger scale. Twelve very large test-tubes were caused to pass air-tight through a slab of wood; the wood was thickly coated with cement, in which, while it was hot and soft, a heated ‘ propagating glass,’ resembling a huge bell-jar, was imbedded. The air within the glass was pumped out several times, air filtered carefully through 88 THE FLOATING-MATTER OF THE AIR. a plug of cotton-wool being permitted to supply its place. The test-tubes contained infusions of hay, tur- nip, beef, and mutton, three of each, twelve in all. For two months they remained as clear and cloudless as they were upon the day of their introduction, while twelve similar tubes, prepared at the same time, in pre- cisely the same way, and hung on to the slab of wood outside the propagating-glass, were, in less than a week, clogged with mycelium, mould, and Bacteria. One of the protected tubes was accidentally broken, and though its aperture was rapidly plugged with cotton-wool, some common air must at the time have entered the propagating-glass. Evaporation from the infusions went on; the vapour was condensed by the glass above, trickled down its interior surface, carrying with it, in part, such matter as had attached itself to that surface. A kind of pool was thus formed upon the cement below. This after an interval of three months became spotted with disks of Penicilliwm, by the spores of which one or two of the infusions have been recently invaded, the production of very beautiful mycelium- tufts being the consequence. § 19. Hxperiments with Caleined Air. Six years ago' I showed that the floating matter of London air could be removed by permitting a platinum wire heated to whiteness to act upon it for a sufficient time. I availed myself of this mode of calcining the air on the present occasion. The apparatus employed is shown in fig. 5. A glass shade, 8, is placed upon a slab of wood mounted on a tripod. Through the slab passes three large test-tubes nearly filled with the in- fusion to be examined. A platinum spiral, p, unites ' Proc. Roy. Inst. vol. vi. pp. 4 and 5. PUTKEFACTION AND INFECTION. 89 the ends of two upright copper wires, which pass through the stand and are seen coiled outside it. The shade is surrounded below by a tin collar, with a space of about half an inch all round between it and the shade. This space is filled with cotton-wool firmly packed. Connecting the wires with a battery of fifteen cells, the spiral p was raised to whiteness, and was permitted to continue so for five minutes. Experiments previously executed had shown that this sufficed for the entire removal of the floating matter. When the spiral was heated, a portion of the expanded air was driven through the cotton - wool packing below; and when the current was interrupted, this air, returning into the shade, was prevented by the cotton-wool from carrying any floating matter with it. The first three substances brought into contact with air calcined in this way were damson-juice, pear-juice, and infusion of yeast. They were boiled for five min- utes, and for five months they have remained without speck or turbidity. Other tubes similarly boiled, and placed underneath shades containing the floating matter of the air, have long since fallen into mould and rotten- ness. Turnip- and hay-infusions, rendered slightly alkaline, have been mentioned as particularly prone to spon- taneous generation. I wished to test this. On the 26th of November, therefore, four shades were prepared, 90 THE FLOATING-MATTER OF THE AIR. two containing strong turnip-infusion and hay-infu- sion unneutralized, two containing the same infusions slightly superneutralized by caustic potash. The al- leged spontaneous development of life was not observed. The tubes exhibit to this hour the clearness and colour which they showed on the day they were boiled. Her- metically-sealed tubes, containing the same infusions, prepared on the same day, remain equally clear; while the specimens exposed to the laboratory air have fallen into rottenness. The experiments with calcined air were also executed in another form and on a larger scale. A ‘ propagating- glass,’ similar to that already described, was cemented in the same way to a slab of wood through which passed twelve large test-tubes. The infusions, as before, were hay, turnip, beef, and mutton. The air being removed from the propagating-glass by a good air-pump, its place was supplied by other air which had passed slowly through a red-hot platinum tube containing a roll of platinum gauze, also heated to redness. Tested by a searching beam, this calcined air was found quite free from floating matter. For two months no speck invaded the limpidity of the infusions exposed to it, while a week’s exposure to the ordinary air sufficed to reduce twelve similar infusions, hung on to the slab of wood outside the glass, to the muddiness of putrefaction. § 20. Infusions withdrawn from Air. The arrangement here was the same as that adopted in the first experiment with filtered air, the only dif- ference being that the bell-jar, with a view to its more perfect exhaustion, was smaller. It was cemented air- tight to a slab of wood through which passed three large test-tubes, filled to about two-thirds of their PUTREFACTION AND INFECTION. 91 capacity with infusions of beef, mutton, and turnip re- spectively. The bell-jar was exhausted six times in succession, and filled after each exhaustion with air carefully filtered through cotton-wool. While this air was in contact with the infusions they were boiled in a brine-bath. The receiver was afterwards exhausted as perfectly as a good air-pump could exhaust it; while outside the receiver were hung three tubes to compare with those within. Here the protected infusions remained as clear as they were on the day of their introduction, not only after the exposed infusions had charged themselves with life, but for many weeks after they had evaporated away. Such, then, are the tests to which I have subjected the statement that ‘boiled turnip- and hay-infusions exposed to filtered air, to calcined air, or shut off al- together from contact with air, are more or less prone to swarm with Bacteria and Vibriones in the course of from two to six days.’ These results, and others that might be adduced, leave no doubt upon my mind that the deportment of air from which the floating matter has been removed by filtration or calcination is precisely the same as that of air from which the particles have disappeared by self-subsidence. Once really sterilized, an infusion in .contact with optically pure air, however obtained, remains sterile. § 21. The Germ-theory of Contagious Disease. It is in connection with the germ-theory of con- tagious disease that the doctrine of spontaneous genera- tion assumes its gravest aspect. My interest in the general question was first excited by the investigations of Pasteur, while the medical bearings of the doctrine 92 THE FLOATING-MATTER OF THE AIR. were subsequently made clear to me, mainly, I ought to say, by the writings and conversation of the late Dr. William Budd, who was the first of our countrymen to grasp with true philosophic insight the doctrine of ‘the vitality of contagia,’ which is now every day gain- ing ground. At the present moment, indeed, no other medical principle occupies so much thought, or is the subject of so much discussion. ‘How does it happen,’ says Dr. Burdon Sanderson!, ‘that these Bacteria, which we suppose must have existed half a dozen years ago in as great numbers as at present, were then scarcely heard of, and that they now occupy so large a place in the medical literature of this country and of Germany, and have lately afforded material for lively discussion in the French Academy?’ Dr. Sanderson points out the re- lation of Lister in England, and of Hallier in Germany, to the movement regarding Bacteria which is now work- ing likea ferment through the medical world. But to no other workers in this field are we more indebted than to Dr. Sanderson himself, and to his colleagues, for the continued and successful prosecution of re- searches bearing upon the pathology of contagion. ‘In 1870, writes Mr. John Simon, in one of his excellent reports to the Privy Council, ‘I had the honour of presenting Dr. Sanderson’s first. report of researches made in this matter. At that time general conclusions seemed justified, first, that the characteristic-shaped elements which the microscope had shown abounding in various infective products are self-multiplying organic forms, not congeneric with the animal body in which they are found, but apparently of the lowest vegetable kind; and, secondly, that such living organisms are probably the essence, or an inseparable part of the 1 British Medical Journal, Janusry 16, 1875. PUTREFACTION AND INFECTION. 93 essence, of all contagia of disease. . . . This view of the matter has since then become greatly more dis- tinct, in consequence of the investigations made by Dr. Sanderson, particularly in 1871 and 1872, with refer- ence to the common septic contagium or ferment. For in that ferment there seems now to be identified a force which, acting disintegratively upon organic matter, whether dead or living, can, on the one hand, initiate putrefaction of what is dead, and, on the other hand, initiate febrile and inflammatory processes in what is living.’ At a Meeting of the Pathological Society, held on the 6th of April, 1875, the germ-theory of disease was formally introduced as a subject for discussion, the debate being continued with great ability and earnest- ness at subsequent meetings. The Conference was attended by many distinguished medical men, some of whom were profoundly influenced by the arguments, and none of whom disputed the facts brought forward against the theory on that occasion. The leader of the debate, and the most prominent speaker, was Dr. Bastian, to whom also fell the task of replying on all the questions raised. The coexistence of Bacteria and contagious disease was admitted; but, instead of con- sidering these organisms as ‘ probably the essence, or an inseparable part of the essence’ of the contagium, Dr. Bastian contended that they were ‘ pathological products,’ spontaneously generated in the body after it had been rendered diseased by the real contagium. The grouping of the ultimate particles of matter to form living organisms, Dr. Bastian considers to be an operation as little requiring the action of antece- dent life as their grouping to form any of the ‘ other less complex chemical compounds.’ Such a position must, of course, stand or fall by the evidence which its 94 THE FLOATING-MATTER OF THE AIR. supporter is able to produce; and accordingly Dr. Bastian appeals to the law and testimony of experiment as demonstrating the soundness of his view. He seems quite aware of the gravity of the matter in hand; this is his deliberate and almost solemn appeal :—‘ With the view of settling these questions, therefore, we may carefully prepare an infusion from some animal tissue, be it muscle, kidney, or liver; we may place it in a flask whose neck is drawn out and narrowed in the blow- pipe-flame, we may boil the fluid, seal the vessel during ebullition, and, keeping it in a warm place, may await the result, as I have often done. After a variable time the previously neated fluid within the hermetically sealed flask swarms more or less plentifully with Bacteria and allied organisms—even though the fluids have been so much degraded in quality by exposure to the tempera- ture of 212° Fahr., and have thereby, in all probability, been rendered far less prone to engender independent living units than the unheated fluids in the tissnes would be.’ ! We have here, to use the words of Dr. Bastian, ‘a question lying at the root of the pathology of the most important and most fatal class of diseases to which the human race is liable.’ Let us now examine his settle- ment of the question, as described by himself in the foregoing extract. § 22. Experiments with Hermetically-sealed Vessels. Experiments with hermetically-sealed tubes were begun by me on the 5th of October, 1875. The shape of the tubes after sealing is represented in fig.6. Each of them contained about an ounce of liquid. They ! Transactions of the Pathological Society of London, 1875, p. 272. PUTREFACTION AND INFECTION. 95 were boiled for only three minutes in an oil-bath, and were sealed, during ebullition, not by a blowpipe, but by the far more effectual spirit-lamp flame. Hay.—Four tubes were charged on the date mentioned with a strong infusion, four with a weak infusion. All eight flasks remain to the present hour clear. Turnip.—tTwo kinds of turnip were tried in these first experiments. Two tubes were charged with a strong infusion, and two with a weak infusion of a sound hard turnip; while two other pairs of tubes were filled with strong and weak infusions from a soft woolly turnip. All the tubes remain transparent to the present time. Two or three days’ exposure to the air of the laboratory sufficed to cloud all these infusions and fill them with life. On the 8th of October twenty-one tubes were charged with infusions of the following substances :— Mackerel, beef, eel, oyster, oatmeal, malt, potato. There were three tubes of each infusion. All of them remain to the present hour unchanged. I had not previously seen a more beautiful illustra- tion of the dichroitic action which produces the colours of the sky than in the case of the oyster-infusion. With reflected light it presented a beautiful cerulean hue, while it was yellow by transmitted light. This was due to the action of suspended particles which defied alike the power of the microscope and of ordinary fil- tration. At right angles to a transmitted beam the infusion copiously discharged perfectly polarized light. Fie. 6, 96 THE FLOATING-MATTER OF THE AIR. Suspended particles in the potato-infusion produced a somewhat similar effect, but it was by no means so fine as that of the oyster-infusion. By ordinary filtra- tion it was not possible completely to rid the malt and oatmeal infusions of suspended matter; but both remain exactly as they were when the flasks containing them were sealed. These experiments had been made before the volume of the Transactions of the Pathological Society con- taining the discussion referred to above came into my hands,.' It caused me to turn again to my tubes, seek- ing further evidence. Onthe 12th of November thirty- six of them were charged, boiled, and hermetically sealed ; on the 13th fifty-seven, on the 16th thirty-one, and on the 17th six tubes were similarly treated. The entire group of tubes, therefore, numbered one hundred and thirty. I tried moreover to multiply the chances of spontaneous generation by making the infusions of the most diverse materials. The following Table gives ‘the names of the substances operated on, the number of tubes sealed, and the date of sealing :— Fowl tubes. | November 12th. Mutton . Wild Duck Beef ; Herring . Haddock . Mullet Codfish Pheasant . Rabbit Hare Snipe ” ” DDADAADAAAAAAaIwWR S 1 To the courtesy of Dr. Bastian I am indebted for a separate copy of the report of the discussion here referred to. PUTREFACTION AND INFECTION. 97 Partridge. Plover Liver Heart Tongue of Sheep Brains of Sheep Sweetbread . Humour of Ox-eye (anata) Lens of Ox-eye , Lungs of Sheep Tripe Sole. tubes November 13th. ” ” ” ” » 55 November 16th. ” ” APAMWNAWA QR AA 7 S ” a November 17th. The tubes were immersed in groups of six at a time in an oil-bath, boiled for three minutes, and then sealed. More than one hundred of these flasks were sensibly transparent and free from turbidity at the outset, and they remain so to the present hour. In some cases, however, it was not possible to wholly remove turbidity by filtration. Ihave already referred to the opalescence of oyster-infusion, which has invariably appeared when- ever oyster has been digested. A still more pronounced case of the kind is furnished by an infusion of the crystalline lens of the ox. Nothing hitherto encountered by me imitates the flush of the true opal so closely as this infusion. Filtration through one hundred layers of paper was quite incompetent to remove the suspended particles to which this opalescence is due. Some of the other infusions remained turbid after filtration, without ex- hibiting what I should call opalescence. The sheep’s lungs furnish an example of this. In some cases, moreover, where repeated filtering failed to remove the suspended particles, a few weeks’ quiet caused them to sink, and leave the supernatant liquid clear. It may be worth remarking that some rabbit-infusions have shown a decided opalescence, while others have heen 98 FLOATING-MATTER OF THE AIR. perfectly clear. The same remark applies to turnip- infusions, some of which have been found as clear as dis- tilled water, while in general a slight opalescence is not to be got rid of by filtering. These later experiments are quite in harmony with the earlier ones. Not a single flask of this multitude manifests the deportment alleged to be a matter of common observation.’ If the power of spontaneous generation be a scientific verity, surely amid opportu- nities so multiplied and various it must have exerted itself. That the infusions employed were not ‘de- graded’ by the boiling so as to be incapable of sup- porting life, was proved by the fact that exposed tubes containing the same infusions, treated in precisely the same way, resolved themselves with the usual speed into Bacterial swarms, § 23. Conditions as to the Temperature and Strength of Infusions. In connection with these experiments, I have sought, to the best of my ability, to meet every condition and requirement laid down by others as essential to success. With regard to warmth, a temperature of 90° was generally attainable in our laboratory, while on cer- tain days of mild weather without, and in favourable positions within, the temperature to which the infu- sions were subjected reached over 100° Fahr. As Dr. Bastian, however, had laid considerable stress on warmth, thougn most of his results were obtained with temperatures from 15° to 30° lower than mine,? I 1 This group of flasks was submitted to the inspection of the Fellows of the Royal Society on the 13th of January, 1876. 2 Proc. Roy. Soc. vol. xxi. p. 130. Also ‘ Beginnings of Life,’ vol. i. p. 354. PUTREFACTION AND INFECTION, 99 thought it desirable to meet this new requirement also. The sealed tubes, which had proved barren in the Royal Institution, were suspended in boxes copiously per- forated, so as to permit of the free circulation of warm air, and placed under the supervision of an intelligent assistant in the Turkish Bath in Jermyn Street. The washing-room of the establishment was found to be particularly suitable for our purpose; and here, accord- ingly, the boxes were suspended. From two to six days are allowed by Dr. Bastian for the generation of organisms in hermetically-sealed tubes. Mine remained in the washing-room for nine days. Thermometers placed in the boxes, and read off twice or three times a day, showed the temperature to vary from a minimum of 101° to a maximum of 112° Fahr. At the end of nine days the infusions were as clear as at: the beginning. They were then removed to a warmer position; the temperature 115° having been mentioned as favour- able to spontaneous generation. For fourteen days my temperatures hovered about this point, falling once as low as 106°, reaching 116° on three occasions, 118° on one, and 119° on two. The result was quite the same as that recorded a moment ago. The higher temperatures proved perfectly incompetent to develop life.! Fifty-six observations, including both the maximum and minimum thermometers, were taken while the tubes occupied their first position in the washing-room, and seventy-four while they occupied the second position. The whole record, carefully drawn out, is before me, but I trust the statement of the major and minor limits of temperature will suffice. Dr. Bastian’s demand for these high temperatures is, as already remarked, quite recent. Prior to my com- 1 My thanks are due to the managers of the bath for their obliging kindness in this matter. 100 THE FLOATING-MATTER OF THE AIR munication to the Royal Society on January 13, he had successfully worked with temperatures lower than those within my reach in Albemarle Street. There I followed his directions, adhered strictly to his prescriptions; bul, taking care to boil and seal the liquids aright, his results refused to appear in my experiments. On learning this “he raised an objection as to temperature, and made a new demand. With this I have complied ; but bis posi- tion is unimproved. With regard to the question of concentration, I have already referred, in sections 3 and 16 of this memoir, to the great diversity in this particular presented by all my infusions, through their slow evaporation. But more than a general conformity to prescribed con- ditions was observed here also. The strength of an infusion is regarded as fixed by its specific gravity ; and I have worked with infusions of precisely the same specific gravity as those employed by Dr. Bastian. This I was specially careful to do in relation to the experiments described and vouched for, I fear incan- tiously, by Dr. Burdon Sanderson in vol. vii. p. 180 of ‘Nature.’ It will there be seen that, though failure attended some of his efforts, Dr. Bastian did satisfy Dr. Sanderson that in boiled and hermetically-sealed flasks Bacteria sometimes appear in swarms. With purely liquid infusions I have failed to reproduce this result. Hay- and turnip-infusions, of accurately the same character and strength as those employed on the occasion referred to, were prepared, boiled in an oil-bath, carefully sealed up, and subjected to the proper temperatures. In multiplied experiments they remained uniformly sterile.' 1 One hundred and twenty flasks, hermetically sealed, contain- ing animal and vegetable infusions, some two, some three yerrs old, are now beside me. They show no sign of Bacterial life. PUTREFACTION AND INFECTION. 101 § 24. Developmental Power of Infusions and Solu- tions: Air-germs contrasted with Water-germs. Wishing to make no experiment, whether with self- cleansed, filtered, or calcined air, or with infusions with- drawn from air by the air-pump, or contained in hermetically-sealed vessels, without exposing the same infusions to ordinary air, this comparison was instituted with the substances mentioned at pages 96 and 97. One hundred test-tubes, an inch wide and 3 inches deep, were divided into groups, each group being filled with the same infusion. The groups were sufficiently numerous to embrace all the substances mentioned in the Table referred to. Exposed to the uncleansed air, they were attacked with different degrees of rapidity and vigour; but in a few days all of them without exception became muddy and crowded with life. On the whole, the hare- and pheasant-infusions presented the greatest contrast. The tubes containing the former were far gone before those containing the latter were sensibly invaded. The putrescibility of pheasant, more- over, was exceeded by that of snipe, partridge, and plover. The sheep’s heart examined was also slow to putrefy. A single illustration of this difference of developmental power may be given here. On the 13th of November thirty tubes, containing infusions of partridge, pheasant, snipe, hare, sheep’s heart, and codfish, five tubes being devoted to each, to- gether with four tubes of plover, three of mullet, and three of liver, were exposed to the laboratory air. On the 15th, 16th, and 22nd the numbers taken possession of by Bacteria were as follows :— 102 THE FLOATING-MATTER OF THE AIR. 15th. 16th. 22nd, Partridge . : : . 0 3 all Pheasant : 0 1 oe Snipe . Poe) 3 os Hare . . 2 4 i Heart . . 0 1 as Codfish a 2 4 39 Plover. a i 2 ” Mullet . il 2 55 Liver . rae 3 55 They had doubtless all given way some days before the 22nd, but I had not taken the precaution to look at them. Thus, then, the first two days produced no visible change in the pheasant-infusion, while in two of the hare-tubes putrefaction had vigorously set in. Three days’ exposure caused only one of the pheasant-tubes to yield ; four of the hare-infusion had yielded in the same time. The difference between them was also illustrated by the mould upon their surfaces. Some days after their exposure four of the five pheasant-tubes were thickly covered with Penicillium, while the five hare- tubes, with one exception, which could hardly be con- sidered such, had repelled that enemy, maintaining their Bacteria undisturbed. Still the deportment of the hare-infusion may have been due, not to any specific difference between hare and pheasant, but to the circumstances preceding death. The researches of Dr. Brown-Séquard show that even the same animal tissue exhibits, under different circum- stances, very different tendencies to putrefaction. In guinea-pigs subjected immediately after death to the action of the magneto-electric current, he found the rapidity of putrefaction to correspond with the violence of the tetanization. He also draws attention to the PUTREFACTION AND INFECTION. 103 influence of muscular exercise on cadaveric rigidity and putrefaction, showing how quickly they appear in ‘ over- driven cattle and in animals bunted to death. It is known, indeed, to sportsmen that a shot hare will remain soft and limp for a day, while a hunted one becomes rigid in an hour or two. In September, 1851, two sheep which had been overdriven to reach a fair were killed by the section of the carotid arteries. ‘ Putrefac- tion,’ says Dr. Brown-Séquard, ‘ was manifest before the end of the day, or in less than eight hours after death.’ ! The deportment of the hare operated upon by me may therefore depend upon the circumstance of its being brought down by the greyhound instead of the gun. It will be interesting to inquire how far the peculiarity of the animal tissue is transferred to the infusion. This is a fit subject for further investigation.? Such observations inculcate caution in drawing in- ferences from the deportment of any infusion as to the distribution of germs in the air. The germs may be demonstrably present while the infusion may not favour their development. As to the quantity and quality of at- mospheric germs, the hare and the pheasant might lead to different conclusions. A passing reference to an im- portant practical inference may be fitly introduced here. In one of the earliest of the able series of researches with which he has enriched medical science, Dr. Burdon Sanderson exposed to the air ‘Pasteur’s solution,’ which is capable of vigorously developing and nourishing Bacteria 1 Croonian Lecture, Proc. Roy. Soc. 1862, vol. xi. p. 210. 2 Five-and-twenty flasks containing pheasant-infusion were compared during the month of December with five-and-twenty containing infusion of hare, Neither in the rapidity of Bacterial development, nor in the readiness to support the growth of Peni- cillium, did the considerable differences between hare and pheasant first observed repeat themselves. 6 104 THE FLOATING-MATTER OF THE AIR. when they are communicated to it by inoculation; he also permitted air to bubble through the liquid, and find- ing no development in either case, he inferred the entire absence of Bacteria and their germs from the air, con- sidering water to be their exclusive habitation. Other distinguished men have come to the same conclusion ; while in his books and papers, and in the discussion be- fore the Pathological Society already referred to, Dr. Bastian has forcibly dwelt upon the result as justifying the interpretation which he has affixed to his experi- ments. If, he rightly urges, the air be ‘ entirely free’ from matter which could produce Bacteria, then their appearance in boiled infusions exposed to the air must be due, not to anything contained in the air, but to the inherent power of the infusions. Spontaneous genera- tion is undoubtedly the logical outcome of the position that ‘the germinal matter from which Bacteria spring does not exist in ordinary air.’ The experiments, how- ever, recorded in this memoir constitute an ocular demonstration of the respective parts played by the infusion and the air. A pinch of fungus-spores, taken between the fingers, sown in a suitable medium, and producing their appropriate crop, could not more clearly indicate the origin of that crop than experiments with the luminous beam indicate the origin of our harvests of Bacteria. Dr. Sanderson is, I doubt not, now well aware that his first statement was founded on an error of interpretation. In a lecture delivered at Owens College, Manchester, and published in the ‘British Medical Journal’ for January 16, 1875, he to a great extent qualifies and corrects his first inference. He there says that the Bacteria ‘attach themselves without doubt to these minute particles, which, scarcely visible in ordinary light, appear as motes in the sunbeam, or in the beam of an electric lamp.’ In fact the experiments on which PUTREFACTION AND INFECTION. 105 he based his first inference owed their barrenness, not to the absence of Bacteria-germs from the air, but to the inability or, rather, slowness of his mineral solution to develop them. With regard to the part played by the visible motes, I may repeat here what has been previously stated, namely, that while the coarser particles could hardly exist in their midst without loading themselves to some extent with the minute germs of Bacteria, there is no reason to think the motes indispensable for the diffusion of the germs. Whether they are attached to each other or not, the dryness and the moisture of the air are shared equally by both. The germs, moreover, float in the air more readily than the larger particles ; and they, I doubt not, when properly illuminated, shed forth a portion of that changeless light to which reference has been already made, and the perfect polarization of which declares the smallness of the masses which scatter it. The prevalence of the germinal matter of Bacteria in water has been demonstrated by the experiments of Dr. Burdon Sanderson. But the germs in water, it ought to be remembered, are in a very different condi- tion, as regards readiness for development, from those in air. In water they are already wetted, and ready, under the proper conditions, to pass rapidly into the finished organism. In air they are more or less desiccated, and require a period of preparation more or less long to bring them up to the starting-point of the water-germs.' 1 The process by which an atmospheric germ is wetted would be an interesting subject of investigation. A dry microscope covering-glass may be caused to float on water for a year. A sewing-needle may be similarly kept floating, though its specific gravity is nearly eight times that of water. Were it not for some specific relation between the matter of the germ and that of the liquid into which it falls, wetting would be simply impossible. Antecedent to all development there must be an interchange of 106 -THE FLOATING-MATTER OF THE AIR. The rapidity of development in an infusion infected by either a speck of liquid containing Bacteria, or by a drop of distilled water, is extraordinary. On the 4th of January I dipped a thread of glass almost as fine as a hair into a cloudy turnip-infusion, and introduced the tip only of the glass fibre into a large test-tube containing an infusion of red mullet: twelve hours subsequently the perfectly pellucid liquid was cloudy throughout. A second test-tube containing the same infusion was in- fected with a single drop of the distilled water furnished by Messrs. Hopkin and Williams; twelve hours also sufficed to cloud the infusion thus treated. Precisely the same experiments were made with herring infusion, with the same result. In the winter season several days’ exposure to warmed air are needed to produce this effect with air-germs. On the 31st of December a strong infusion was pre- pared by digesting turnip in distilled water at a tem- perature of 120° Fahr. It was divided between four large test-tubes, in one of which the infusion was left unboiled, in another boiled for five minutes, in the two remain- ing ones boiled and, after cooling, infected with one drop of beef-infusion containing Bacteria. In twenty- four hours the unboiled tube and the two infected ones were cloudy, the unboiled tube being the most turbid of the three. The infusion in the unboiled tube was pecu- liarly limpid after digestion; for turnip it was quite exceptional, and no amount of searching with the micro- scope could reveal in it at first the trace of a living Bacterium; still germs were there which, suitably nourished, passed in a single day into Bacterial swarms without number. Five days failed to produce an effect matter between the germ and its environment; and this inter- change must obviously depend upon the character of the encom- passing liquid. PUTREFACTION AND INFECTION. 107 approximately equal to this in the uninfected boiled tube, which was exposed to the common laboratory air. There cannot, I think, be a doubt that the germs in the air differ widely among themselves as regards pre- paredness for development. Some are fresh, others old; some are dry, others moist. Infected by such germs, the same infusion would require different lengths of time to develope Bacterial life. And this remark, I doubt not, applies to the different degrees of rapidity with which epidemic disease affects different people. In some the hatching-period, if I may call it such, is long, in some short, the differences depending upon the different degrees of preparedness of the contagium.! § 25. Diffusion of Germs in the Air. During the earlier observations recorded in this essay, and others not here mentioned, about 100 ex- posed tubes or flasks had been distributed irregularly in the rooms where the inquiry is conducted. They ex- panded to nearly 1000 in the end: not one of them escaped infection. A few days always sufficed to cloud the exposed infusions, and fill them with Bacterial life. I placed tubes at various points in the Royal Institu- tion—on the roof of the house outside, in my bedroom, in an upper kitchen, in my study, in the upper and lower libraries, in the theatre, model-room, reading-room, managers’ room, and in the kitchen at the bottom of the house below the level of Albemarle Street. All were smitten with putrefaction, and with its invariable 1 The medical student of the future will probably connect these remarks with the following statement of Dr. Murchison :—-‘In that protean disease typhoid fever, I have repeatedly had occasion to observe a remarkable similarity in the course, and even in the complications, according to the source of the poison.’—Trans. Path, Soc. vol. xxvi. p. 315. 108 THE FLOATING-MATTER OF THE AIR. associate, Bacteria. In the rooms without fires the action was slower than in the warmer rooms; but all the infusions gave way in the end. In view of the statements which had been made regarding the scantiness of Bacteriu-germs in the air, observations outside of London would, I thought, be interesting. Accordingly, on the 27th of October, a tube containing an infusion of beef was placed in the hands of Mr. Darwin, who had the kindness to set it in his study at Down and observe its changes. In three days it became cloudy and peopled with Bacteria. The same result was obtained in the open air. Mr. Francis Darwin was good enough to expose an infusion for me in his father’s orchard: the weather was cold, and the progress, therefore, slow; but the tube which had been exposed on the 2nd of November was cloudy and full of Bacteria on the 9th. In Sir John Lubbock’s study a similar result: was obtained. From Sherwood, near Tunbridge Wells, infusions of fowl and wild duck were returned to me by Mr. Siemens thickly turbid and crowded with Bacteria. From Pembroke Lodge, Richmond Park, Mr. Russell returned tubes of turnip, beef, and mutton swarming with life. An infusion of beef exposed at Heathfield Park, Sussex, for a week was returned to me by Miss Hamilton muddy and filled with Bacteria. From Greenwich Hospital] Mr. Hirst sent me tubes of beef-, mutton-, and turnip- infusion filled with vigorous Bacteria. Dr. Hooker was good enough to take charge of three sets of tubes at Kew, each set embracing beef, mutton, and turnip. One set was placed in the conservatory, with a tem- perature of 45° to 50°; one in his own study, with a temperature of 54° to 60°; a third set was placed in the orchid-house (the hottest in the gardens), with a temperature of 62° to 75°. PUTREFACTION AND INFECTION. 109 The tubes were exposed on the 4th of December, all of them being then clear. In the orchid-house the turnip became cloudy on the 7th, the beef and mutton on the 8th, after which the opacity rapidly increased. In the study all remained clear until the 9th, when the turnip began to cloud. On the 11th the beef was still clear, while the mutton had given way. On the 13th all of them had yielded. In the conservatory the turnip began to cloud on the 10th; the others followed in the same order as in the other cases. The influence of temperature seems well shown by these observations. Three days sufficed to cloud the turnip in the orchid-house, five days in the study, and six days in the conservatory. The mutton in the study gathered over it a thick blanket of Penicillium. On the 13th it had assumed a light brown coleur, ‘as if by a faint admixture of clay;’ but the infusion became transparent. The ‘clay’ here was the slime of dormant or dead Bacteria, the cause of their quiescence being the blanket of Penicillium. I found no active life in this tube, while all the others swarmed with Bacteria. From the Crystal Palace at Sydenham Mr. Price sent me tubes of mutton, beef, and turnip charged with Bacteria. The temperature was low at night, the development of life being thereby considerably retarded. Thus, wherever it has been tested, the atmosphere has been found charged with the germs of Bacteria. I wished, however, to obtain clearer and more defi- nite insight as to the diffusion of atmospheric germs. Supposing a large shallow tray to be filled with a suitable organic infusion and exposed to the ‘air. Into it the germs would drop; and could the resulting organisms be confined to the locality where the germs fell, we should have the floating life of the atmosphere mapped, 110 THE FLOATING-MATTER OF THE AIR. so to speak, in the infusion. But in such a tray the organisms would intermingle and thus mar the revela~ tion of their distribution. Valuable information I thought might be gained by breaking up the infusion into isolated conterminous patches, and exposing them to the air. A square wooden tray was accordingly pierced with one hundred circular apertures; into each of which was dropped a test-tube 3 inches long and 1 inch wide, with its rim resting in each case upon the rim of the aperture. There were ten rows of tubes, with ten tubes in each row. On the 23rd of October, 1875, thirty of these tubes were filled with an infusion of hay, thirty- five with an infusion of turnip, and thirty-five with an infusion of beef. The tubes with their infusions had been previously boiled ten at a time in an oil-bath. One hundred circles were marked upon paper so as to form a plan of the tray, and every day the state of each tube was registered upon the corresponding circle. Seven such maps or records were executed. I will use the term ‘cloudy’ to denote the early stage of turbidity, distinct but not strong. The term * muddy’ will be used to denote thick turbidity. § 26. Tray of one hundred Tubes. On the 25th of October one or two of the tubes exposed on the 23rd showed signs of yielding; but the progress of putrefaction was first registered on the 26th. Fig. 7, embracing the first record, is annexed; it may be thus described. Hay.—Of the thirty specimens exposed, one had become ‘muddy ’—the seventh in the middle row, reckoning from the side of the tray nearest a stove. Six tubes remained perfectly clear between this muddy one and the stove, proving that differences of warmth PUTREFACTION AND INFECTION. 111 may be overridden by other causes. Every one of the other tubes containing the hay-infusion showed spots of mould upon the clear liquid. Turnip.—F our of the thirty-five tubes were very muddy, two of them being in the row next the stove, one four rows distant, and the remaining one nine rows away. Besides these, seven tubes had become clouded. There was no mould on any of the tubes. Beef.—One tube of the thirty-five was quite muddy, in the seventh row from the stove. There were three cloudy tubes, while seven of them bore spots of mould. As a general rule organic infusions exposed to the air during the autumn remained for two days or more perfectly clear. Doubtless from the first germs fell into them, but the germs required time to become or~ ganisms. This period of clearness may be called the ‘period of latency ;’ and, indeed, it exactly corresponds with what is understood by this term in medicine. Towards the end of the period of latency the fall into a state of disease, if I may use the term, is comparatively sudden ; the infusion passing from perfect clearness to cloudiness more or less dense in a few hours. Thus the tube placed in Mr. Darwin’s possession was clear at 8.33 a.m. on the 19th of October, and cloudy at 4.30 p.m. Seven hours, moreover, after the first record of our tray of tubes, a marked change had occurred. For the purpose of comparison the second record, fig. 8, is placed beside the first. The change may be thus described:—Instead of one, eight of the tubes containing hay-infusion had fallen into uniform rouddiness. Nineteen of them had produced Bacterial slime, which had fallen to the bottom, every tube con- taining the slime being covered by mould. ‘Three tubes only remained clear, but with mould upon their sur- faces. The muddy turnip-tubes had increased from four to ten; seven tubes were clouded, while eighteen of them 112 THE FLOATING-MATTER OF THE AIR. remained clear, with here and there a speck of mould on the surface. Of the beef, six were cloudy and one thickly muddy, while spots of mould had formed on the majority of the remaining tubes. Fifteen hours subsequent to this observation, viz. on the morning of the 27th of October, all the tubes containing hay-infusion were smitten, though in different degrees, some of them being much Fic, 7. 26th Oct., noon. oe@Qd0aceE COO®OOGOOO solelelelelel_lelele elejeje!_leleiciele. OOOOCO8OOE Jelelele) lle) _) ol Yelele\ lelel1 | QOQDODOOOGO IQOOOLYOODO SOBG®OGHOO O © © © Mould, Cloudy. Muddy. more iehag than others, Of the turnip-tubes, three only remained unsmitten, and two of these had mould upon their surfaces. Only one of the thirty-five beef- infusions remained intact. A change of occupancy, moreover, had occurred in the tube which first gave way. Its muddiness remained grey for a day and a half, then it changed to bright yellow-green, and PUTREFACTION AND INFECTION. 113 maintained this colour to the end. On the evening of the 27th every tube of the hundred was smitten, the majority with uniform turbidity, some, however, with mould above and slime below, the intermediate liquid being clear. The whole process bore a striking resem- blance to the propagation of a plague among a popula- tion, the attacks being successive and of different Fi, 8, 26th Oct., 7 Px. O@000HDe8® O0SO00GOGE: OO' = OW} ®) © QOOOVSOGO® OO S0@6660 ii ( dINUAL AVH GQOIOOO V@VO@OO OOGOO2O V@GEE@O0O V@QO@OGDO QOQOOOS@09O 0EVO0O V@@OO6O © @ J 2 a 5 Ee =) a a 4 © OQ. Or®@ Slime. Slime and Mould. Clear. degrees of virulence. I annex copies of the fourth and seventh maps (figs. 9, 10) with their respective dates. On the 31st of October I finally inspected the tray of tubes. All those containing the hay-infusion were turbid, some thicker and much more deeply coloured 114 THE FLOATING-MATTER OF THE AIR. than others. They had been all at first aliké in colour. Out of the thirty tubes four only were free from mould. Three of these were adjacent to each other, the fourth at a distant portion of the tray. The Penicillium was exquisitely beautiful. Its prevalent form was a circular patch made up of alternate zones of light and deep green. In some cases the liquid Fig. 9. 27th Oct., 6.30 P.M. COD OOOQ90C6 16 @60000008 0060806060 COOCOBOSSS2 O08 6606660 62602808660 é S97 90809 5068008 oS Q Cloudy. Green. oO © Slime, Slime and mould. Clear. was covered by a single large patch; in others there were three or four patches, each made up of its dif- ferently coloured zones. Reticulated patterns also occurred, Three kinds of Penicillium seemed strug- gling for existence, namely:—that just described; a second kind, of the same consistency and colour, fat PUTREFACTION AND INFECTION. 115 forming little rounded heaps instead of circles ; thirdly, a woolly, voluminous, white mould, in the middle of which a zoned circle of the other mould sometimes formed a little islet. All the tubes containing the turnip-infusion were also turbid on the 3lst. Nine of them were free from mould. This, where it occurred, exactly resembled Fia. 10, 29th Oct., 10.30 aa, @eececeeece @000000009: small cocoons in shape. The beef-tubes were also all turbid on the 31st, and seventeen of them were free from mould. The mould upon the beef, moreover, was muck. less luxuriant than that on the hay- and turnip- infusions. The mould-developing power is obviously greatest in the hay-, less in the turnip-, and least of all in the beef-infusion. In every case where the mould 116 THE FLOATING-MATTER OF THE AIR. was thick and coherent the Bacteria died, or became dormant, and fell to the bottom as a sediment. The growth of mould and its effect on the Bacteria are very capricious. The turnip-infusion, after developing in the first instance its myriadfold Bacterial life, fre- quently contracts mould, which stifles the Bacteria and clears the liquid all the way between the sediment and the scum. Of two tubes placed beside each other, one will be taken possession of by Bacteria, which success-~ fully fight the mould and keep the surface perfectly clean ; while another will allow the mould a footing, the apparent destruction of the Bacteria being the con- sequence. This I have proved to be the case with all infusions, fish, flesh, fowl, and vegetable. At the pre- sent moment, for example, of three tubes containing an infusion of sole, placed close together in a row, the two outside ones are covered by a thick tough blanket of mould, while the central one has not a single speck upon its surface. The Bacteria which manufacture a green pigment appear to be uniformly victorious in their fight with the Penicillium. These observations enable us, I think, to draw some interesting conclusions. From the irregular manner in which the tubes are attacked we may infer that, as regards quantity, the distribution of the germs in the air is not uniform. A single tube will sometimes be a day or more in advance of its neighbours. The singling out, moreover, of one tube of the hundred by the par- ticular Bacteria that develop a green pigment, and other cases just adverted to, shows that, as regards qual- ity, the distribution is not uniform. This has been further illustrated by the following observations :—Of five-and-twenty tubes of different animal infusions ex- posed in groups of five, in the middle of November, and PUTREFACTION AND INFECTION. 117 all swarming with Bacterial life, five were green. They were distributed as follows:—Beef 2, herring 1, had- dock 1, fowl 1, wild duck 0. The same absence of uniformity was manifested in the struggle for existence between the Bacteria and the Penicillium. In some tubes the former were triumphant; in other tubes of the same infusion the latter was triumphant. It would also seem that a want of uniformity as regards vital vigour prevailed. With the self-same infusion the motions of the Bacteria in some tubes were exceedingly languid; while in other tubes the motions resembled a rain of projectiles, being so rapid and violent as to be followed with difficulty by the eye. Reflecting on the whole of this, I conclude that the germs float through the atmosphere in groups or clouds, and that now and then a cloud specifically different from the prevalent ones is wafted through the air. The touching of a nutritive fluid by a Bacterial cloud would naturally have a dif- ferent effect from the touching of it by the sterile air between two clouds. But, asin the case of a mottled sky, the various portions of the landscape are succes- sively visited by shade, so, in the long run, are the various tubes of our tray touched by the Bacterial clouds, the final fertilization or infection of them all being the consequence.’ 1 In hospital practice the opening of a wound during the pass- age of a Bacterial cloud would have an effect very different from the opening of it in the interspace between two clouds. Certain caprices in the behaviour of dressed wounds may possibly be ac- counted for in this way. Under the heading ‘Nothing New under the Sun,’ Professor Huxley has lately sent me the following remarkable extract :— ‘ Uebrigens kann man sich die in der Atmosphire schwimmenden Thierchen wie Wolken denken, mit denen ganz leere Luftmassen, ja ganze Tage vollig reinen Luftverhiltnisse wechseln.’ (Ehbren- berg, ‘Infusionsthierchen,’ 1838, p. 525.) The coincidence of phraseology is surprising, for I knew nothing of Ehrenberg’s con- ception. My ‘ clouds,’ however, are but small miniatures of his, 118 THE FLOATING-MATTER OF THE AIR. The tray of tubes proved so helpful in enabling me to realize mentally the distribution of germs in the air, that on the 9th of November, 1875, I exposed a second tray containing one hundred tubes filled with an infusion of mutton. On the morning of the 11th six of the ten nearest the stove had given way to putrefaction; three of the row most distant from the stove had yielded, while here and there over the tray particular tubes were singled out and smitten by theinfection. Of the whole tray of one hundred tubes, twenty-seven were either muddy or cloudy on the 11th. Thus, doubtless, in an infective atmosphere, are individuals successively struck down. On the 12th all the tubes had given way, but the differences in their contents were extraor~ dinary. All of them contained Bacteria, some few, others in swarms. In some tubes they were slow and sickly in their motions, in some apparently dead, while in others they darted about with rampant vigour. These differences are to be referred to differences in the ger- minal matter, for the same infusion was presented everywhere to the air. Here also I imagine we have a picture of what occurs during an epidemic, the difference in number and energy of the Bacterial swarms resem- bling the varying intensity of the disease. It becomes obvious from these experiments that of two individuals of the same population exposed to a contagious atmo- sphere, the one may be severely, the other lightly attacked, though, as regards susceptibility, the two individuals may be as identical as two samples of one and the same mutton-infusion. What I have already said regarding the ‘ preparedness’ of contagium has its application here. _ The parallelism of these actions with the progress of infectious disease may be traced still further. The ‘ Times,’ for example, of January 17, 1876, contained a letter on typhoid fever, signed ‘ M.D.,’ in which occurs PUTREFACTION AND INFECTION. 119 the following remarkable statement :—‘ In one part of it [Edinburgh], congregated together and inhabited by the lowest. of the population, there are, according to the Corporation return for 1874, no less than 14,319 houses or dwellings—many under one roof, on the “ flat” system—in which there are no house connexions what- ever with the street-sewers, and, consequently, no water- closets. To this day, therefore, all the excrementitious and other refuse of the inhabitants is collected in pails or pans, and remains in their midst, generally in a par- titioned-off corner of the living-room, until the next day, when it is taken down to the streets and emptied into the Corporation carts. Drunken and vicious though the population be, herded together like sheep, and with the filth collected and kept for 24 hours in their very midst, it is a remarkable fact that typhoid fever and diphtheria are simply unknown in these wretched hovels.’ The analogy of this result with the behaviour of our infusions is perfect. On the 30th of last November, for example, a quantity of animal refuse, embracing beef, fish, rabbit, hare, was placed in two large test-tubes opening into a protecting-chamber containing six tubes. On December 13, when the refuse was in a state of noisome putrefaction, infusions of whiting, turnip, beef, and mutton were placed in the other four tubes. They were then boiled and abandoned to the action of the foul ‘ sewer-gas’ emitted by their two putrid companions. On Christmas-day, 1875, these four infusions were limpid. The end of the pipette was then dipped into one of the putrid tubes, and a quantity of matter, comparable in smallness to the pock-lymph held on the point of a lancet, was transferred to the turnip. Its clearness was not sensibly affected at the time; but on the 26th it was turbid throughout. On the 27th a speck from the infected turnip was transferred to the whiting ; on the 120 THE FLOATING-MATTER OF THE AIR. 28th disease had taken entire possession of the whiting. To the present hour the beef- and mutton-tubes remain as limpid as distilled water. Just as in the case of the living men and women in Edinburgh, no amount of fetid gas had the power of propagating the plague, as long as the organisms which constitute the true con- tagium did not gain access to the infusions. In the foregoing observations the tubes were arranged in the same horizontal plane; but I also sought to ob- tain some notion of the vertical distribution of the germs in the air of the room. Two trays, each contain- ing 100 tubes, were supported the one above the other in the same frame. The upper tray had all the air be- tween it and the ceiling, a height of about 12 feet, from which the germs might descend upon it; the lower tray was shaded by the upper, a space of only 6 inches exist- ing between them. If the number of germs deposited in the tubes were determined by the air-space above, the upper tray would be the one most rapidly and thoroughly taken possession of. The reverse was the case. As regards the development of Bacterial life, the lower tray was from first to last in advance of its neigh- bour. It is not air-space, then, so much as stillness, that determines the deposition of the germs. The air between the two trays being less disturbed than the general air of the room, the germs were less wafted about, and therefore sank in greater numbers into the tubes of the lower tray. We have here data which will enable us to form a rough notion of the lower limit of the number of germs contained in the room where the experiments were made. The floor of the room measured 20 feet by 15 feet ; its area was therefore 43,200 square inches, and every square inch would afford room for the section of one of our test-tubes. The height of the room is 180 inches; PUTREFACTION AND INFECTION. 121 hence 30 layers of tubes 6 inches apart might be placed one above the other between the floor and ceiling. This would make 1,296,000 tubes. If only a single germ a day fell into each tube, this would be the number of the germs. If the number deposited were one an hour, we should have thirty millions a day sown in the tubes. Probably the average time necessary for infec- tion is very much less than an hour. At all events, 30,000,000 of germs daily would be an exceedingly moderate estimate of the number falling into our thirty layers of tubes. This, moreover, would only be a frac- tion—probably a small fraction—of the germs really present in the air. In his Presidential Address to the British Association at Liverpool, Professor Huxley ven- tured the statement that myriads of germs are floating in our atmosphere. Certain experimenters have rashly ridiculed this statement. In view of the foregoing calculation it, however, expresses the soberest fact. Indeed, taking the word myriad in its literal sense of ten thousand, it would be simple bathos to apply it to the multitudinous germs of our air. § 27. Some Experiments of Pasteur and.their Rela- tion to Bacterial Clouds. Quite recently I had occasion to refresh my memory of Pasteur’s paper published in the ‘ Annales de Chimie’ for 1862. The pleasure I experienced on first reading it was revived by its reperusal. Clearness, strength, and caution, with consummate experimental skill for their minister, were rarely more strikingly displayed than in this imperishable essay. Hence it is that during recent discussions, in which this and other labours of the highest rank met with such scant respect, those in England most competent to judge of the value of scien- 122 THE FLOATING-MATTER OF THE AIR. tific work never lost faith in the substantial accuracy of Pasteur. One striking example of his penetration has an immediate bearing on the conclusion regarding Bacterial clouds, independently drawn by me from the deportment of the tray of one hundred tubes. On the 28th of May, 1860, Pasteur opened, on an uncovered terrace a few metres above the ground, four flasks con- taining the water of yeast. Nothing appeared in any of them until the 5th of June, when a small tuft of mycelium was observed in one of them. On the 6tha second tuft appeared in another flask ; the two remaining flasks continued intact and without organisms. On the 20th of July he opened, in his own laboratory, six flasks containing water of yeast. Tour of them remained per- fectly intact, while two of them became promptly charged with organisms. From these observations Pasteur in- ferred the non-continuity of the cause to which so-called spontaneous generation is due. This inference is quite in accord with the notion of Bacterial clouds suggested by my observations. Pasteur, in fact, sometimes opened his flask in the midst of a Bacterial cloud and obtained life, sometimes in the interspace between two clouds, and obtained no life. Not with a view of repeating this observation, which I had forgotten, but for another reason, I opened on the 6th of January last a number of hermetically-sealed tubes in one and the same room of the Royal Institu- tion. The names of the infusions contained in the tubes, the date of sealing them up, their condition be- fore opening on the 6th, and their appearance six days subsequently are given in the accompanying table. J chose for these observations tubes which coutained a little liquid in their drawn-out portions. In every case the motion of this liquid, when the tube was broken, indicated a violent inrush of air. PUTREFACTION AND INFECTION. 123 Tahision. ca aaa gees 7 Oa ae Grouse Nov. 27th Clear Clear. Sole . ‘ » 17th +3 Turbid. Turnip No. 1 Oct. 5th 55 Penicillium on surface. Turnip No. 2 sat his 5 Clear. Tlay . : ” oo” ” Mycelium at bottom. Wild Duck Nov. 12th ‘5 Turbid. Mutton » oo” Cloudy. Fowl moo” ” Clear. Beef . Fy Sy 45 Mycelium at bottom. Haddock . ube, Eg 55 Clear. Sweetbread y 16th i Mycelium at bottom. Rabbit y 18th +) Clear. oath yo» ” Curdy layer at top. Pheasant . a 755 ” Clear. Mullet yoo” ” ” | Hare. » oo” ” ” Snipe . mo” ” ” Partridge . no” ” ” Plover 5 8 93 Mycelium below. Codfish aye ae A Clear. Kidney Jan, 5th ” Mycelium at bottom. Salmon Dec. 18th = Clear. Whiting ” 9 ” ” Turnip. é : » 29th ” 55 Hay, 4 drops of caustic | Nov, 22nd | Clear with | Mycelium potash sediment at bottom. Hay, 2 drops of caustic rae, Clear Mycelium potash at bottom. Hay, 5 drops of caustic ae Clear with | Clear. potash sediment Hay, 6 drops of caustic $3 Clear with i potash sediment Liver Nov. 30th Clear v Hey . y 18th ” ” ay. 9” ” ” Turnip yo” ” Muddy. 124 THE FLOATING-MATTER OF THE AIR. Thus, out of 31 flasks opened in the same air, 18 remained intact, while 13 were taken possession of by organisms—a fact obviously the same in character as that described by Pasteur. Such experiments demon- strate, if demonstration were needed, that it is not the air itself, or any gaseous or vaporous substance " uniformly diffused through it, but some discontinuous substance floating in it, that is the cause of the infec- tion. Instead of our tubes, let us suppose thirty-one wounds to be opened in the same ward of a hospital; plainly what has occurred with the tubes may occur with these wounds—some may receive the germs and putrefy, others may escape. Helped by the conception not only of germs, but of germ-clouds, the different behaviour of wounds subjected apparently to precisely the same con- ditions will cease to be an inscrutable mystery to the surgeon.! During the course of this inquiry some eminent biologists have been good enough, from time to time, to look in upon my work, and to give me their views 1 «We have ample facts of experiment in our hands,’ said Mr. Knowsley Thornton (Trans. of the Pathological Society, vol. xxvi. p. 313), ‘to show that it is not the gases of the air, or any soluble material in water, but something “particulate ”’ which sets up all the train of changes in an open wound, which may, after the patient has passed through a period of more or less constitutional disturb- ance, end in the healing of the wound, or may end in septicemia and death. This particulate material, then, I believe we have evi- dence enough to prove consists of germs of Bacteria and other low organisms.’ All the evidence points to this conclusion. I may say that I entirely agree with Mr. Thornton in the distinction he draws between germs and developed J?acteria floating in the air. It is, in my opinion, of the very last importance to seize this distinction with clearness. When it is fully realized we shall probably hear less of the arguments against Bacterial contagia founded on the fact that a virus diminishes in strength as the Bactcria multiply. A portion of the energy of the virus consists in its passage from the germ state to that of the finished organism. PUTREFACTION AND INFECTION. 125 regarding the evidential force of the experiments. To Professor Huxley, moreover, I am indebted for under- taking the examination of a number of the hermetically- sealed tubes. Thirty of them were placed in his hands, none of them being regarded as defective. A close examination, however, disclosed in one of them a myce- lium. No faultiness could for a time be discovered in the tube; the sealing appeared to be quite as perfect as that of its sterile fellows. Once, however, on shaking it a minute drop of liquid struck my friend’s face ; and he soon discovered that an orifice of almost microscopic minuteness had been left open in the nozzle of the tube. Through this the common air had been sucked in as the liquid cooled, and hence the contamination. It was the only defective tube of the group of thirty, and it alone showed signs of life. The statement of this fact before the Royal Society, by Professor Huxley, brought to my mind a somewhat similar experience of my own. One morning in Novem- ber I lifted one of the hermetically-sealed tubes from the wire on which it was suspended, and, holding it up against the light, discovered, to my astonishment, a beautiful mycelium at the bottom. Before restoring the tube to its place I touched its fused end and found it cutting sharp. Close inspection showed that the nozzle had been broken off; the common air had entered, and the seed of the mycelium had been sown. Two other instances, one like that observed by Professor Huxley, have since come to light. In one of thema minute orifice remained after the supposed sealing of the tube. The other case was noticed when the tubes were returned from the Turkish bath. One of them contained a luxuriant mycelium. It was noticed that the liquid in this tube had singularly diminished in quantity, and on turning the tube up it was found cracked at the bottom. 126 THE FLOATING-MATTER OF THE AIR. No case of pseudo-spontaneous generation ever occurred under my hands that was not to be accounted for in an equally satisfactory manner. In this inquiry, thus far, I have confined my ob- servations to purely liquid infusions, purposely exclud- ing milk, mixtures of turnip-juice and cheese, and, indeed, mixtures of solids and liquids of all kinds. The next section of the investigation will be devoted to these and kindred subjects; and to it I also postpone the complete examination of pepton, and of the remarkable experiments described by Dr. William Roberts, a small residue of which only I have failed to corroborate. Throughout the whole of this investigation I have had to congratulate myself on the zealous and efficient aid of my excellent assistant, Mr. John Cottrell. His intelligence in seizing my ideas, and his mechanical skill in realizing them, have rendered me admirable service. Without such aid, indeed, so much ground could not have been covered in the time. Royal Institution, 5th April, 1876. It gives me special pleasure to direct attention here to a paper by the Rev. W. H. Dallinger, for an advanced proof of which I am indebted to the courtesy of Dr. Lawson, editor of the ‘Popular Science Review.’ Mr. Dallinger and his colleague Dr. Drysdale are known to have pushed the microscope to its utmost power of per- formance at the present time. Their ‘ Researches into the Life-History of the Monads’ are models of scientitic thoroughness and concentration. Mr. Dallinger’s review of the present position of the doctrine of spontaneous generation, his remarks on Bacterial germs in relation PUTREFACTION AND INFECTION. 127 to the limits of the powers of the microscope, his demonstration that the germs of monads survive in a medium raised to a temperature which destroys the adult, and that precipitated mastic particles like those mentioned in § 16 of this paper are not to be discerned by a magnifying-power of 15,000 diameters, constitute a most interesting and important communication. Nore I. Action of Bacteria upon a Beam of Light. To trace the gradual growth and multiplication of the Bacteria by their action on a beam of light, an infusion of beef was prepared on the 5th of October, 1875, placed in a globular flask of about 50 cubic inches capacity, and put aside with its mouth open to the laboratory air. On the 8th, 9th, 10th, 11th, and 12th similar flasks were prepared and put aside in succession. On the 12th all the flasks were examined by the concentrated electric light. The freshest one showed the track of the beam asa richly- coloured green cone. The green light was unaffected by a Nicol’s prism, which, however, quenched the ordi- nary scattered light and augmented the purity and vividness of the green. It was a case of fluorescence. In the second flask, one day old, the fluorescent beam was in great part masked by the scattered light; the latter, however, could be partially quenched by a Nicol’s prism, the purity of the fluorescence being thus in part restored. Through the third flask, two days, and through the fourth flask, three days old, the track of the beam was-still discernible; through the fifth flask, four days old, it was all but obliterated, while in the sixth flask, seven days old, it was entirely shattered, the turbid medium being filled uniformly with the laterally scat- tered light. Two of these flasks were of a bright yellow-green 7 128 THE FLOATING-MATTER OF THE AIR. colour, two were milky or white, and two of a dull brownish hue. Cohn mentions the bluish tinge of the infusion by reflected and its yellow tinge by transmitted light when the Bacteria are incipient. This is due to a dichroitic action, similar to that which produces the blue of the sky and the morning and evening red. The blue, how- ever, though discernible, is not pronounced, for the Bacteria are too large to scatter the colour in any high degree of purity; but with a ‘muddy’ infusion a very fair red may be obtained from transmitted light. I have used the Bacterial turbidity for photometric put - poses. On the 9th of October, for example, I ac- companied Sir Richard Collinson and a Committee of the Elder Brethren of the Trinity House to Charlton, with the view of comparing together two lights mounted at the Trinity Wharf at Blackwall. To imi- tate a foggy atmosphere, I employed an infusion cloudy with Bacteria and placed in a glass cell. With it the beams could be toned gradually down to complete ex- tinction. Norte II. Fluorescence of Infusions. All the animal infusions, both flesh and fish, showed the same fluorescence. Jt was the same green hue throughout, though of varying degrees of intensity. In wild duck, grouse, snipe, hare, partridge, and pheasant the fluorescence was fine—sometimes exceed- ingly fine. In rabbit it was less fine than in hare, and in a tame rabbit less fine than in a warren rabbit. Fishes also differed from each other. Mullet, for example, was finer than cod, herring, or haddock. Beef, mutton, heart, liver, all showed the same green fluorescence. Led up to it by a series of remarkable experiments PUTREFACTION AND INFECTION. 129 on the rapidity of the passage of crystallized substances into the vascular and non-vascular textures of the body,! Dr. Bence Jones and Dr. Dupré communicated to the Royal Society in 1867 a highly interesting paper ‘On a Fluorescent Substance, resembling Quinine, in Ani- mals,’* They then showed that ‘from every texture of man and of some animals a fluorescent substance can be extracted, which, when extracted, has a very close optical and chemical resemblance to quinine.’ They therefore called it animal quinoidine. In dilute solu- tions they found that the fluorescence of the animal substance was not to be distinguished from that pro- duced by quinine. When the solution was concentrated, the colour of the light was of a decidedly greenish hue. This latter observation is most in agreement with mine. In all the infusions examined by me the fluorescent light was a decided green, and not to be mistaken for the blue light of quinine. The green colour is similar to that emitted by the crystalline lens when a beam of violet light impinges on it; sending such a beam through any of the infu- sions, the ‘degradation’ of the violet to green is strikingly illustrated. The foregoing statement refers to the deportment of the infusions after boiling and filtering. Prior to boiling some of them were of a brilliant ruby colour ; but even here, when the layer of liquid between the eye and the beam was not too thick, the green fluor- escence could be seen through the red liquid. 1 Proceedings of the Royal Society, vol. xiv. 1865. 2 Tbid. vol. xv. p. 73. 3 On plunging the eye into the beam of the electric lamp, trans- mitted through violet glass, the moment the crystalline lens is seen to fluoresce by a second observer, a blue shimmer is seen by the eye on which the beam falls. In the case of my own eye, I can always readily tell when the fluoresence has set in. FURTHER RESEARCHES ON THE DEPORT- MENT AND VITALITY OF PUTREFACTIVE ORGANISMS." i aaa III. § 1. Introduction. On the 13th of January, 1876, I had the honour of submitting to the Royal Society some account of an investigation in which the power. of atmospheric air to produce life in organic infusions and its power to scatter light were shown to go hand in hand. The * scattering ” was proved to be due, not to the air itself, but to foreign matter suspended in the air. It was moreover proved that air placed under proper conditions went through a process of self-purification, and that, when this purification was visibly complete, the power to scatter light and to generate life disappeared to- gether. The form of the experiments here referred to was, it will be remembered, as follows:—Wooden chambers were constructed with glass fronts, side windows, and back doors. Through the bottoms of the chambers test-tubes passed air-tight, their open ends, for about one-fifth of the length of the tubes, being within the chambers. Provision was made fora connexion through sinuous channels between the outer and the inner air. 1 Philosophical Transactions, Part I., 1877. 132 THE FLOATING-MATTER OF THE AIR. The chambers, being closely sealed, were permitted to remain undisturbed for a few days. The floating matter of the internal air gradually subsided, until at length an intensely luminous beam failed to show its Fig. 11, track within the chamber. Then, and not till then, were the infusions introduced, by means of a pipette passing through the top of the chamber. After their introduction, they were boiled in an oil or brine-bath ! 1 From the fact of their being boiled in oil or brine, Professor VITALITY OF PUTREFACTIVE ORGANISMS. 133 for five minutes, and afterwards placed permanently in a warm room. The annexed woodcut (fig. 11), taken from the ‘ Proceedings’ of the Royal Institution, shows a cham- ber with its six test-tubes, its side windows w w, its pipette c, and its bent tubes a b, which connect the air of the chamber with the external air. In upwards of fifty chambers thus constructed, many of them used more than once, it was, without excep- tion, proved that a sterilized infusion in contact with air shown to be self-cleansed by the luminous beam re- mained sterile. Never, in a single unexplained instance, did such an infusion show any signs of life. That the observed sterility was not due to any lack of nutritive power in the infusion, was proved by opening the back door and permitting the uncleansed air to enter the chamber. The contact of the floating matter with the infusions was invariably followed by the development of life. Numerous examples of these results were placed before the Fellows of the Royal Society at their Meet- ing on the 13th of January, 1876. Prior to the date here referred to, great public interest had been excited, and, I may add, considerable scientific uncertainty had been produced in reference to this subject, both in England and America, by the writings of Dr. Bastian. These writings consisted, in part, of theoretic considerations and reflections, not new, but sometimes very ably stated, based on the general doctrine of Evolution; and, in part, of very pungent criticisms of those who, though believers in Evolution, declined to accept the writer’s programme Cohn has inadvertently inferred that the infusions themselves were raised above their boiling-points. The tubes being open, the tem- perature of ebullition is of course independent of the source which provokes it. 134 THE FLOATING-MATTER OF THE AIR. of its operations! Passing over both theory and criticism, I thought it wise to fix upon certain well defined statements of fact which lay at the basis of the weighty superstructure raised by their author, and to bring these statements to the test of strict experiment. Thus it was affirmed ‘that boiled turnip or hay- infusions exposed to ordinary air, exposed to filtered air, to calcined air, or shut off altogether from contact with air, are more or less prone to swarm with Bacteria and Vibriones in the course of from two to six days.’ ? I resorted accordingly to filtered air, calcined air, and to infusions withdrawn from air, but failed to discover the alleged‘ proneness’ to run into living forms. It had also been affirmed that infusions of muscle, kidney, or liver, placed ‘in a flask whose neck is drawn out and narrowed in the blowpipe flame, boiled, sealed during ebullition, and kept in a warm place, swarmed after a . variable time with Bacteria and allied organisms.’! I resorted to such flasks, employing infusions of fish, flesh, fowl, and viscera, and on the 13th of January was able to place before the Royal Society one hundred and thirty flasks, every one of which negatived the fore- going statement. Two objections were subsequently urged against these results. The infusions, it was contended, were not sufficiently concentrated, nor were the temperatures sufficiently high. Both these objections were met by the statement that forty-eight hours’ exposure under the same circumstances to common air sufficed to fill these same infusions with life. Beyond this, however, I was able to show that the temperatures employed by me were exactly those which had previously been found 1 See ‘Evolution, or the Origin of Life,’ pp. 168, 169. 2 « Mvolution,’ p. 94. » Transactions of the Pathological Society, 1875, p. 272. VITALITY OF PUTREFACTIVE ORGANISMS. 135 ‘most effectual by the writer who urged the objection. Other temperatures, higher than any previously em- ployed, were at the same time said to ensure sponta- neous generation. I exposed my infusions to these newly-discovered efficient temperatures, but found that they remained as barren as before. With regard, moreover, to the question of concen- tration, it was shown that, owing to their gradual vaporization, the infusions used by me were probably unequalled in strength by those employed by any pre- vious investigator. Some of these infusions remain with me to the present hour. Concentrated by twelve months’ slow evaporation, and reduced to one-fifth of their primitive volume, they still exhibit the purity of distilled water, These results proved beyond a doubt that in the atmospheric conditions existing in the laboratory of the Royal Institution during the autumn, winter, and spring of 1875-6, five minutes’ boiling sufficed to sterilize organic liquids of the most diverse kinds. Among these may be mentioned urine in its natural condition, infusions of mutton, beef, pork, hay, turnip, haddock, sole, salmon, cod-fish, turbot, mullet, her- ring, eel, oyster, whiting, liver, kidney, hare, rabbit, barn-door fowl, grouse, and pheasant. Once properly sterilized, and protected afterwards from the floating matter of the air, not one of these putrescible infusions ever manifested the power of generating by its own in- herent energy putrefactive organisms of any kind. | § 2. Experiments of Pasteur, Roberts, and Cohn. During the investigation just referred to I confined myself for the most part to animal and vegetable juices in their natural condition—that. is to say, extracted 136 THE FLOATING-MATTER OF THE ATR. by distilled water, and not rendered artificially acid, neutral, or alkaline. I had occasion, however, to repeat among others some of the very remarkable experiments on superneutralized hay-infusions described by Dr. William Roberts in his excellent paper in the Philo- sophical Transactions for 1874. These experiments I could not corroborate; for while in his hands such infusions sometimes required three hours’ boiling to sterilize them, in mine they behaved like other in- fusions, and were sterilized in five minutes. In the abstract of the investigation communicated to the Royal Society on the 13th of January, 1876, I mentioned this discrepancy, and pointed out its possible cause.! But the largeness of the question, which had been long previously raised by M. Pasteur, and the limitation of my time, led me to postpone it. This postponement is mentioned at the conclusion of my paper in the Philosophical Transactions for 1876, where the discrepancy referred to is not at all discussed. In his celebrated paper, ‘Sur les corpuscules orga- nisés qui existent dans l’Atmosphére,’ published fifteen years ago,’ M. Pasteur first announced that while acid infusions had their germinal life destroyed by a tem- perature of 100° C., a temperature over 100° was needed to produce the same effect in alkaline infusions. In his ‘Etudes sur la Biére,’ published in the early part of 1876, he repeats and illustrates this statement. Vinegar he finds has the organisms which decompose it destroyed by a temperature of 50°C. Wine is ren- dered unchangeable by a slightly higher temperature. Beerwort without hops requires a temperature of 90°C. to sterilize it, and milk a temperature of 110°. Fresh urine has its organisms destroyed at a temperature of 1 Roy. Soc. Proc. vol. xxiv. p. 178. * Annales de Chimie, 1862, vol. Ixiv. VITALITY OF PUTREFACTIVE ORGANISMS. 137 100°, while a higher temperature is needed when the urine has been neutralized by carbonate of lime.! The resistance of alkalized urine to sterilization is therefore by no means a new announcement.? On my return from Switzerland in 1876 the experi- ments on alkalized hay-infusions were resumed; and soon afterwards Professor Cohn, of Breslau, so highly distinguished by his researches on Bacteria, placed in my hands a memoir * which rendered it doubly incum- bent on me to examine more strictly the grounds of my dissidence from Dr. Roberts. Professor Cohn is, on the whole, emphatic in his corroboration of Dr. Roberts,‘ having found, during a long and varied series of ex- periments with hay-infusions of divers kinds, that when the period of boiling did not exceed fifteen minutes organisms invariably appeared in the infusions after- 1 «Etudes sur la Biare,’ p. 34. 2 With regard to the different action of acid and alkaline liquids, I put the subject purposely aside with the view to its full investigation as soon as the first instalment of these researches had been published. I could find no adequate explanation of the alleged fact that germs are killed in an acid liquid, while they survive in an alkaline one of the same temperature; nor could the well-merited respect that I feel for M. Pasteur cause me to accept his explanation without further inquiry on my own account. In due time, therefore, I resolved to examine the question. Various experiments and explanatory views regarding it are recorded in the following pages. 3 Beitrige zur Biologie der Pflanzen, July 1876. « Professor Cohn gently censures me for taking exception to the cotton-wool plug, seeing that cotton-wool, even in my own experi- ments, has always proved a trustworthy filter. I did not, however, object to it asa filter, but on grounds which have in part, at all events, commended themselves to Professor Cohn himself. With reference to the method of Dr. Roberts he writes thus :—‘ The de- fect of this method consists in the difficulty of protecting the cotton-wool from accidental wetting by the infusion. The steam, moreover, which rises from the liquid penetrates the cotton-wool, and, through its partial condensation in the neck of the bulb, might readily charge itself with germs.’ 138 THE FLOATING-MATTER OF THE AIR. wards. Sixty, eighty, and even one hundred and twenty minutes’ boiling were found in some cases in- sufficient to sterilize the infusions. One marked dif- ference, however, exists between Dr. Roberts and Pro- fessor Cohn. ‘The former found five minutes’ boiling sufficient to sterilize unneutralized hay-infusion, but one, two, and even three hours’ boiling insufficient to sterilize superneutralized hay-infusion; while the latter noticed no difference of this kind, but found acid and neutral infusions equally resistant. § 3. Hay-infusions. Preliminary Experiments with Pipette-bulbs. I have now the honour to submit to the Royal Society an investigation which embraces among others the points here referred to, and which has proved far more difficult and laborious than I expected it would be. On the 27th of September, 1876, a quantity of chopped hay was digested for three hours and a half in distilled water maintained at a temperature of 120° Fahr. The infusion was afterwards poured off, and its specific gravity reduced to the exact figure given by Dr. Roberts, viz. 1006. It was then filtered and slightly super- neutralized. Precipitation occurred on the addition of the potash, and the infusion was boiled for five minutes to render the precipitation complete. It was then re- filtered, and introduced into a series of bulbs of the same size and character as those described by Dr. Roberts, and called by him ‘ plugged bulbs.’? Each bulb was a cylinder about four inches high and upwards of an inch wide, with a long neck attached to 1 «Kin constanter Unterschied in der Zeitdauer zwischen sauren und neutralen Aufgiissen, wie ihn Roberts gefunden, trat in unseren Versuchen nicht hervor ’ (p. 259). 2 Phil. Trans. vol. clxiv. p. 460. VITALITY OF PUTREFACTIVE ORGANISMS. 139 it,! as shown at a, fig. 12, Two-thirds of the cylinder were occupied by the infusion. After the introduction of the latter, the neck of the bulb was plugged with cotton-wool, and hermetically sealed above the plug, as at B, fig. 12. The bulbs were afterwards plunged in water deep enough to cover their necks. The water was Fig. 12. gradually raised to the boiling-point, and maintained at the boiling temperature for ten minutes. They 1 JT have called them ‘pipette-bulbs’ because they are formed by hermetically sealing one shank of a pipette, close to the bulb, leaving the other shank open for the introduction of the infusions. German pipettes, on account of their cheapness, were at first com- monly used; but in cases of long-continued boiling, explosions were so frequent that bulbs of English glass of specially resistant quality were resorted to. 140 THE FLOATING-MATTER OF THE AIR. were then removed and permitted to cool; after which the sealed end of each neck was broken off by means of a file, its subsequent appearance being shown at c, fig. 12. The bulbs, protected by the cotton-wool plugs in the neck above them, were then exposed to a tolerably uniform temperature of about 90° Fahr. At the same time two similar bulbs, charged with the same infusion, had their necks bent downwards, Fic, 13. as in fig. 13, the inclined por- tion being plugged, so that no impurity could fall into the liquid from the cotton-wool. These two bulbs were boiled for five minutes in an oil-bath, X33, and plugged while boiling with cotton-wool. They were then sealed behind the plugs and permitted to cool, their sealed ends being broken off after- wards. On the 30th of September the infusion in all the straight- necked bulbs was turbid, while in the two bent-necked ones it was perfectly clear. On the 2nd of October the turbidity of the straight-necked bulbs had in- creased, while a fatty scum had formed on the sur- face of each. The two others were at the same time slightly but distinctly turbid. My inference from this experiment was that in neither the straight-necked nor the bent-necked bulbs bad the germs been wholly killed by the boiling. The difference between the results obtained with the re- spective bulbs arises from the different modes of mani- VITALITY OF PUTREFACTIVE ORGANISMS. 141 pulation pursued in the two cases. For it is to be aoted that a quantity of air, with its associated floating. matter, was imprisoned above the infusion in every straight-necked bulb; that in the case of the two bent- necked bulbs this air had been in part displaced by steam, the air which entered on cooling being sifted by the cotton-wool plugs. To this difference of treatment is to be attributed the observed difference of deport- ment. Unlike the thick cloudiness of their neigh- bours, the turbidity of the bent-necked bulbs, though distinct, was barely sensible, and in none of them was any scum ever formed upon the surface of the infusion. Examined microscopically, numerous Vibrios were found in the infusions of the straight-necked bulbs, many of them broken at the centre, with the two halves apparently trying to separate from each other. There were also numerous smaller Bacteria, very active and of various lengths. In the bent-necked bulbs a num- ber of exceedingly small Bacteria were found, but no Vibrios. The deportment of the hay-infusion employed in these experiments corroborates the results of Dr. Roberts and Professor Cohn. On the 2nd of October another infusion of hay was prepared, and, after neutralization with caustic potash, was introduced into six pipette-bulbs with straight necks. The necks, being first plugged with cotton- wool, were afterwards sealed by the blowpipe. The infusions were maintained for ten minutes at the tem- perature of boiling water. Their sealed ends were afterwards broken off, and they were subjected, like the former ones, to a temperature of 90° Fahr. Six other bulbs were charged at the same time with the same infusion; but, instead of being hermetically sealed, they were placed in an oil-bath, and boiled there 142 THE FLOATING-MATTER OF THE AIR. for five minutes. Before the ebullition ceased, the neck of each was stopped with a plug of cotton-wool. Up to October 6th all the bulbs continued clear. On the 6th one bulb of the series last described be- came turbid, lighter in colour than its neighbours, and covered with a fatty scum. On the 7th one tube of the first series (boiled after the fashion of Roberts for ten minutes) also became turbid and exhibited the same fatty scum. The remaining ten bulbs maintained per- manently their deep brown-sherry colour, their high transparency, and their perfect freedom from Bacterial life. They are still clear, though seven months have elapsed since their preparation. In the great majority of these experiments the de- portment of alkalized hay-infusion contradicts that observed by Dr. Roberts and Professor Cohn. Six other pipette-bulbs, with their necks so bent and plugged with cotton-wool and asbestos that no impurity falling from the plug could reach the infusion, were also charged on the 2nd of October. Three of the bulbs, with their necks hermetically sealed, were main- tained for ten minutes at the temperature of boiling water, the sealed ends being afterwards broken off. The three other bulbs were boiled in an oil-bath, and had their necks plugged before ebullition ceased. All six bulbs have remained perfectly transparent up to the present time. Here, again, we have discordance between my re- sults and those of Dr. Roberts and Professor Cohn. But on the 6th of October another infusion was prepared and neutralized, exactly in the same fashion as before. Five pipette-bulbs were charged with it; they were hermetically sealed and maintained at the boiling temperature for ten minutes. The sealed ends were afterwards broken off, and the bulbs exposed to 2 VITALITY OF PUTREFACTIVE ORGANISMS. 143 temperature of 90° Fahr. On the morning of the 8th of October (that is to say, two days after their pre- paration) the infusion in every one of the bulbs was turbid and covered with scum. Here once more we have perfect harmony between my results and those of Dr. Roberts and Professor Cohn. On the 2nd of October, moreover, fourteen of our ordinary small retort-flasks with bent necks (shown in fig. 14) were charged with the neutralized hay-infusion. They were boiled for three mi- nutes, and hermetically sealed whilst boiling. Some days after- wards one tube of the entire number was observed to have be- come lighter in colour and sen- sibly cloudy; but thirteen out of the fourteen remained unchanged in colour, brightly transparent, and entirely free from life. Here the dissidence between my results and those of Professor Cohn, who also experimented with her- metically-sealed flasks, reappears. Numerous other experiments with pipette-bulbs and retort-flasks were made at the time here referred to, but it is unnecessary to record them. Suffice it to say that, like those just described, some of them corrobo- rated and some of them contradicted the results of Dr. Roberts and Professor Cohn. Fie. 14. 144 THE FLOATING-MATTER OF THE AIR. § 4. Hay-infusions. Experiments with Cohn’s Tubes. For reasons given by himself,! Professor Cohn de- viated from the method of experiment pursued by Dr. Roberts, employing, instead of the pipette- bulbs, flasks, the nature of which will be under- stood from the following description. Let a zone of a common test-tube, about one-third of its length from its open end, be softened by heat, and let the softened glass be drawn out so as to form a tube of much narrower bore than the original test-tube. Thus modi- fied, the tube would consist of an elongated bulb below and an open funnel above, both being connected, by a narrow neck (see fig. 15). Professor Cohn filled the elongated bulb to about two-thirds of its volume with hay- infusion, plunged his bulbs in water, raised the water to ebullition, and continued the boiling for the required time. The tubes were then removed from their bath, and after being held open for a minute or two so as to allow the water condensed in their necks to evaporate, the funnel was plugged with cotton-wool. Professor Cohn considers that all possibility of external contamination is here shut out.2 By his method, therefore, I wished to check the results above 1 Beitriige, July 1876, p. 256. 2 ¢Bhe ich itiber die Organismen berichte, welche sich in den gekochten Aufgiissen entwickelten, will ich bemerken, dass an eine nachtrigliche Infection derselben durch von aussen nach dem Kochen eingeschleppte Keime bei unseren Versuchen nicht zu denken ist’ (p. 259). I may remark that, with an atmosphere like that in which my recent experiments were conducted, there would be no chance of escape for an infusion thus handled. VITALITY OF PUTREFACTIVE ORGANISMS. 145 described. Accordingly, on the 24th of October, I had four groups of Cohn’s tubes (twelve in a group) carefully charged with two fresh infusions of two different kinds of hay. Each infusion was divided into two equal parts, one of which was neutralized and the other left in its natural acid condition. Twelve of the tubes were charged with one of the infusions neutralized, and twelve with the same infusion unneutralized. We will label this in- fusion A. Twelve other tubes were charged with the second infusion neutralized, and twelve with it unneu- tralized. We will call this infusion B. The forty-eight tubes were subsequently boiled for ten minutes in tin vessels containing water deep enough nearly to submerge them. Having proved by previous experiments that it was dangerous if not fatal to exactness to expose the in- fusions for one or two minutes to the air after their removal from the water, I took the precaution of plug- ging them first and removing them afterwards. On the 28th of October (that is to say, four days after their preparation) several of the tubes containing the unneutralized infusion A were faintly but distinctly turbid and thinly covered with scum. The twelve neutralized tubes of the same infusion were at the same time perfectly clear. This retarding influence of the alkali has been of frequent occurrence in this inquiry. That it was simply a case of retardation was proved by the fact that, on the 30th of October, the twenty-four tubes, both neutral and acid, of infusion A were tur- bid and covered with scum. On the same date the twelve neutralized tubes of infusion B were perfectly clear and without a trace of scum. Of the twelve unneutralized tubes three had given way, and a fourth yielded on the 31st. Four days later three of the neutralized tubes also yielded. The permanent state of matters was that eight out of 146 THE FLOATING-MATTER OF THE AIR. the twenty-four tubes charged with infusion B had become turbid, while sixteen of them remained perfectly clear. I do not doubt that the tardy infection of some of the tubes just referred to arose from external con- tamination, which is almost inseparable from the method of experiment. Here, while infusion A corroborated Professor Cohn, infusion B in substance contradicted him. § 5. Hay-infusions (in Closed Chambers). In dealing with hay-infusions I also fell back on the method of experiment which was found so effectual in 1875,' employing closed chambers in which the air had been permitted to cleanse itself by the gradual subsi- dence of its floating matter. On the 3rd of October, 1876, my experiments with such chambers recommenced. Two of them, containing. each three large test-tubes, were then charged with an infusion of hay accurately prepared according to the prescription of Dr. Roberts. Its specific gravity was 1006; it was superneutralized to the proper extent with caustic potash, but the period of boiling, instead of being three hours, was five minutes. Examined from time to time for more than four months subsequently, the infusion in both chambers continued perfectly unchanged. It was free from sus- pended matter, free also from every trace of scum, main- taining for the light which passed through it a singular transparency. Here, to a certainty, a period of boiling not amount- ing to one-twentieth of that required by Dr. Roberts, sufficed to destroy totally the power of generating life in an alkalized hay-infusion. 1 Briefly described in the Introduction, VITALITY OF PUTREFACTIVE ORGANISMS. 147 This result is in perfect harmony with all the results of last year. Chamber after chamber was then charged with infusions of hay, which were afterwards subjected to the boiling temperature for five minutes. In every chamber the infusion remained perfectly clear until purposely infected from without. There was no instance observed last year in which five minutes’ boiling failed to sterilize hay-infusion, whether neutralized or un- neutralized. Thus, on the 26th of November, 1875, a group of three test-tubes was charged with hay-infusion of the same specific gravity and of the same degree of alkalinity as that found most resistant by Dr. Roberts. They were protected by glass shades, the air within the shade being calcined by an incandescent platinum wire in the man- ner described in the last essay.! The tubes were boiled for five minutes, the subsequent intrusion of contami- nated air being prevented by a ring of cotton-wool. Thirteen months afterwards the infusion, greatly con- centrated by evaporation, exhibited its pristine deep transparency. A second similar group of tubes was charged with alkalized hay-infusion on the 27th of last January, and on the 5th of December (that is to say, after a period of more than ten months) the infusion was found perfectly clear. A number of hermetically-sealed tubes charged with the same infusion, and boiled for only three minutes, have maintained for more than a year both their primi- tive transparency and their water-hammer sound. Thus many of the earliest experiments of the present year, and the whole body of last year’s experiments, are in complete harmony with each other. This harmony was, however, disturbed by some of the foregoing experiments with bulbs and tubes, and it 1 Phil, Trans., vol. clxvi. p. 50. 148 THE FLOATING-MATTER OF THE AIR. was soon to be further disturbed by experiments with closed chambers. On the 6th of October, 1876, for ex- ample, an infusion was got ready in strict imitation of that prepared on the 3rd; it was of the same specific gravity, it was alkaline to the same degree, and it was , introduced in the same manner into a chamber of three tubes; but whereas the infusion of the 3rd remained intact for months, and would have remained so indefi- nitely, a week had not elapsed before every tube of this new infusion was turbid and covered with fatty scum. § 6. Desiccation of Germs. New Hay and old. In his work entitled ‘Evolution, and the Origin of Life,’ Dr. Bastian affirms, with repeated emphasis, that living matter is unable to maintain its life when exposed to a temperature even below that of boiling water. He refers to the scalding of the hand and other destructive effects, and also to the action of boiling water on eggs. He also refers to the experiments of Spallanzani on seeds, and extends the results observed with living matter of these special kinds, to living matter generally. ‘It has been shown,’ he writes,' ‘ and is believed by the great majority of biologists, that the briefest exposure to the influence of boiling water (212° F.) is destructive of all living matter.’ More than ten years ago an extremely significant observation directly bearing upon this subject was made by the wool-staplers of Elbceuf, in France. They were accustomed to receive dirty fleeces from Brazil, and among other matters entangled in the wool were the seeds of a certain plant called Medicago. It had been repeatedly found by the wool-cleaners that these seeds sometimes germinated after a period of four hours’ 1 ¢ Evolution,’ p. 46, VITALITY OF PUTREFACTIVE ORGANISMS. 149 boiling. The late M. Pouchet repeated the experiment. He collected the seeds, boiled them for four hours, and sowed them afterwards in proper earth. To his aston- ishment they proved fruitful. He then closely examined the boiled seeds, and found the great majority of them swollen and disorganized; but amongst these ruined seeds he observed others which had refused to imbibe the water or to swell or break up in any way. These he carefully picked out, and sowed them and their neigh- bours separately in the same kind of earth. The swollen seeds were incapable of germination, while the unaltered ones rapidly gave birth to a crop. This was the only instance of such resistance known to Pouchet when he communicated the fact to the Paris Academy of Sciences. The observation here described stands recorded in the ‘ Comptes Rendus’ for 1866, vol. lxiii. p. 939. It is not difficult, indeed, to see that the surface of a seed or germ may be so affected by desiccation and other causes as practically to prevent contact between it and a sur rounding liquid.!’ The body of a germ, moreover, may be so indurated by time and dryness as to resist power- fully the insinuation of water between its constituent molecules. It would be difficult to cause such a germ to imbibe the moisture necessary to produce the swelling and softening which precede its destruction in a liquid of high temperature. In my last’ paper I made some remarks upon this subject ;? and in relation to our present experiments, 2 In this connexion a remark of Dr. Roberts regarding the re- sistance of chopped green vegetables merits quotation. ‘The singu- lar resistance of green vegetables to sterilization appears to be due to some peculiarity of the surface, perhaps their smooth glistening epidermis, which prevented complete wetting of their surfaces.’ 2 Phil. Trans., vol. clxvi. p. 60. 150 THE FLOATING-MATTER OF THE AIR. the influence of drying and hardening was brought home to me by the fact that in all the foregoing cases the infusions which five minutes’ boiling proved sufficient to sterilize were, without exception, derived from fresh hay mown in 1876, while the infusions which five minutes’ boiling failed to sterilize were derived, with- out exception, from old hay mown either in 1875 or some previous year. In the earlier experiments of the present inquiry this distinction between old and new hay came most clearly and definitely out. The result was subsequently blurred by circumstances which it required time and labour to unravel, and which will require patience on the reader’s part if he would thoroughly follow them. They will, however, throw far more light upon the rea! character of these inquiries, and do more to reconcile the discords to which researches on spontaneous genera- tion have given birth, than if every experiment had been a success unshaded by doubt. § 7. Hay-infusions. Further experiments with Closed Chambers. With a view to probing to the uttermost this ques- tion of drying and hardening, on the 6th of October an extensive series of experiments with closed chambers was begun. Three different kinds of hay were em- ployed:—lst, Old hay, sent to me by Lord Claud Hamilton, from Heathfield, Sussex;! 2nd, new hay from Heathfield (both, it may be stated, from a some- what ungenerous soil); 3rd, new hay purchased in London, and artificially dried for some days upon a sand- 1 After the possible influence of hard drying and hardening had suggested itself, I purposely introduced old hay from various locali- ties into the laboratory. VITALITY OF PUTREFACTIVE ORGANISMS. 151 bath. For these experiments eleven closed chambers were prepared, as I wished every result to be based as far as possible upon the testimony of two chambers. On the 6th of last October they were carefully charged with the infusions, the period of boiling afterwards being five minutes. Two chambers were devoted to the acid and two to the alkalized infusion of old hay. Two chambers were also devoted to the acid and two to the alkalized infu- sion of dried hay. Two chambers were finally devoted to the alkalized and one to the natural acid infusion of new Heathfield hay. Examined from day to day, differences were svon observed, not only between the different infusions, but also between different chambers containing the same infusion. Thus every tube of both the chambers con- taining the neutralized infusion of old hay became turbid, but the three tubes of the one chamber were loaded in four days with a fatty scum, while the tubes of the other chamber remained for ten days perfectly free from scum. The two chambers containing the acid infusion of old hay exhibited similar differences. Every tube in both of them became turbid; but in one of them the infusion was scumless throughout, while in the other each of the three tubes was heavily laden with scum. The two chambers containing the alkalized infusion of dried London hay had all their tubes turbid and covered with scum. In the case of the acid infusion of dried hay, the tubes of one of the chambers became turbid, while the tubes of the other chamber remained clear. : The two chambers of alkalized new Heathfield hay- infusion were also in disaccord. In the one chamber all three tubes became turbid and covered with scum, 8 152 THE FLOATING-MATTER OF THE AIR. while in the other chamber the three tubes remained sensibly clear and free from scum. Nor did the three tubes of the single chamber charged with the new Heathfield acid ififusion present the same appearance ; for while one tube became thickly turbid, the other two remained perfectly pellucid. Amid this confusion, the only point worth dwelling on is; that while no single case of escape occurred with the old-hay infusion, whether acid or neutral, with the infusions of both dried and undried new hay a certain percentage of the tubes remained sterile. Reflection on these results naturally drew suspicion upon the chambers. They had been used before, and, though carefully cleansed, some unobserved source of infection may have clung to them. This, at all events, seemed the most rational way of accounting for the differences observed between samples of the self-same infusion placed in different chambers. Hence my desire to expose a fresh series of infusions in chambers which had never been used before. Six new ones were therefore constructed, each of them containing six tubes. These were charged on the 3rd of November with infusions of old London hay, old Heathfield hay, new London hay, and dried London hay. Two chambers were devoted to each infusion, which in the one chamber was neutralized and in the other unneutralized. The six tubes in each chamber were arranged in two rows of three tubes each. Those nearest to the glass front were called the front tubes, the others the back tubes. The infusion intended for the unneutralized chamber was unboiled before its introduction into the three back tubes, and boiled in those tubes for five minutes afterwards: the infusion for the front tubes was VITALITY OF PUTREFACTIVE ORGANISMS. 153 boiled for fifteen minutes before introduction and for five minutes afterwards. These differences in the mode and period of boiling were adopted to ascertain whether they had any influence on the subsequent development of life. In the case of the neutralized chambers, the infusion for the three back tubes was boiled for fifteen minutes outside before neutralization, and five minutes in the chamber after neutralization. The infusion for, the three front tubes was boiled fifteen minutes outside after neutralization, and five minutes afterwards in the chamber. If the potash used for neutralization carried germs into the infusion, the difference between five and twenty minutes in the period of boiling might, it was thought, declare itself in the subsequent phenomena. Four days after its introduction the old Heathfield acid infusion was found turbid throughout and covered with scum. The scum and turbidity were sensibly the same in all the tubes, though the period of boiling varied from five to twenty minutes. On the same day the neutralized infusion of the same hay was perfectly brilliant and free from scum. Three days subsequently, however (that is to say, on the 10th of November), the neutralized tubes also became turbid and covered with scum. The salient, fact here to be noted is, that in neither the neutral nor the acid chamber did a single tube of the old Heathfield hay-infusion maintain its primitive clearness and freedom from scum. The old London hay behaved substantially as the old Heathfield hay, no single tube escaping either in the neutralized or the unneutralized chamber. The dried new London hay comes next. A week after its introduction every one of the six tubes con- taining the acid infusion was turbid and coated with scum. In the neutralized chamber, on the contrary, 154 THE FLOATING-MATTER OF THE AIR. two only of the back tubes gave way, the third back tube and the three front tubes remaining clear. On the 8rd of November, moreover, a new chamber of six tubes was charged with an infusion of new London hay. Three of the tubes were neutralized and three unneutralized. Both infusions were introduced into the chamber unboiled, and were boiled afterwards for five minutes. Ina week all the tubes had given way, be- coming turbid in the same degree and covered to the same extent with scum. The newness of the hay had failed to secure the sterility of the infusions. Nothing of this kind occurred in the experiments of last year. It was then found that hay-infusions of all kinds were uniformly sterilized by five minutes’ boiling. Guided by such hints as the experiments furnished, I continued to work. On the 4th of November four closed chambers of three tubes each were charged with infusions of old and new Heathfield hay—two chambers with the one, and two chambers with the other. One chamber of each pair contained a neutral- ized, the other an unneutralized infusion, and the time of boiling was ten minutes. Six days subsequently the infusion of new hay, both neutralized and unneutralized, was found perfectly unchanged. Of the old-hay in- fusion, on the other hand, only one of the six tubes escaped. The three acid tubes became completely tur- bid, while two out of the three neutral ones fell into the same condition. § 8. Expervments with Soaked Hay. Pondering still further on the influence of drying and hardening, and recognizing the necessity of not only wetting but also softening the germs, the thought VITALITY OF PUTREFACTIVE ORGANISMS. 155 occurred to me of soaking the hay for some days prior to digesting it. Old London hay was accordingly chopped up and placed in three glass vessels—one con- taining distilled water, another acidulated water, and a third alkalized water. The superior extractive power of the alkalized liquid was at once manifest ; it rapidly assumed a dark colour. The distilled water came next, yielding a colour less deep than that of the alkalized, but more deep than that of the acidulated water. The alkaliné and distilled-water infusions emitted a rich odour of hay, while the smell of the acid infusion was very faint, and not like that of hay. The hay was per- mitted to soak from the 8th to the 11th of November. It was then digested for three hours in the same liquid at a temperature of 120° F., boiled, filtered, and intro- _ duced into the closed chambers, where it was reboiled in each case for five minutes. Prior to digesting the hay in the liquid in which it had been soaked, Bacteria had developed in swarms. These, of course, were killed by the boiling, and they were not entirely removed by the filtration. The alka- line infusion, indeed, though filtered repeatedly, was sufficiently turbid to prevent the flame of a candle placed behind the tubes containing it from being seen. The same to a less extent was true of the distilled- water infusion. This latter had been divided into two portions, one of which was accurately neutralized, and the other left unneutralized, a separate chamber being devoted to each. From the 11th to the 18th of November the only change observed in any of the infusions was in the direction of increased transparency. They all became clearer with time, the distilled-water infusions becoming particularly clear and brilliant at the top. After two or three days’ quiet the alkaline infusion allowed a flame 156 THE FLOATING-MATTER OF THE AIR. placed behind it to be seen of a deep red. The acidu- lated-water infusion remained entirely unchanged ; but this is not worth dwelling on, for in this case, even when exposed to the common air, the infusion resisted infection for a considerable time. In no case was the fatty scum which had been already so frequently observed formed in any one of the tubes. Some change inimical to the particular organ- isms which produce this scum must have been caused by the soaking of the hay. Examined microscopically on the 18th of November these infusions, I thought, exhibited undoubted evi- dences of Bacterial life. Bacterial forms were unques- tionably there in considerable numbers, more particularly in the sediment at the bottoms of the tubes. Nor dol now see any valid grounds for doubting the presence of life; but I was warned against drawing too hastily the conclusion which first prompted itself, by boiling an in- fusion swarming with active Bacteria, and submitting the liquid after cooling to microscopic examination. Here also the dead Bacterial forms were preserved, and it was extremely difficult to distinguish their motions, which were certainly Brownian motions, from those ob- served in the protected infusions of soaked hay. The experiment was thought worth repeating. On the 16th of November accordingly chopped bundles of old Heathfield hay and new Heathfield hay, and of old London hay and new London hay, were placed in glass dishes containing distilled water, and were thus soaked until the 18th. They were then moved from the lower laboratory, and taken, with their glass covers, to a dis- tant room at the top of the Royal Institution. Here the four specimens of hay were digested for three hours at a temperature of 120° Fahr. They were filtered, boiled, refiltered, some of them through 100 layers of VITALITY OF PUTREFACTIVE ORGANISMS. 157 filter-paper ; after which they were introduced into four closed chambers of six tubes each, and then boiled for five minutes. On the 20th of November. the infusions in all the chambers appeared to be as free from organisms as at first. The new Heathfield and the new London hay- infusions in their respective chambers had their some- what turbid columns surmounted by an exceedingly clear zone of liquid, due, I should consider, to the me- chanical subsidence of the particles, had vot subsequent experience taught me to regard this appearance as a sign of life. On the 23rd scum had begun to gather on every tube of the case containing the infusion of old Heath field hay. On the 30th this scum continued, but there was no trace of it in any of the chambers containing new Heathfield hay, new London hay, and old London hay. These infusions were all somewhat turbid ; but the turbidity differed very little from that exhibited when the infusions were prepared. I spent a good deal of time over these infusions of soaked hay, both with the microscope and otherwise, but the recorded observations would not add materially to our knowledge. I therefore dismiss them with the remark that their general drift was in favour of the idea that the extraordinary resistance to sterilization mani- fested by the old-hay infusions is the result of hardening and desiccation. The foregoing observations, however, have been noted, more with the view of indicating my line of thought than of claiming for them any value whatever as a demonstration. 158 THE FLOATING-MATTER OF THE AIR. § 9. Infusions of Fungi. Turning from hay to substances in which germs, if they existed, could not be desiccated, I felt pretty sure that infusions of such substances would be unable to resist the boiling temperature. To test the correctness of this view the following experiments were made :— Three different kinds of fungi (red, black, and yellow) were gathered in Heathfield Park on the 13th of Octo- ber, and digested separately in London on the following day. Three tubes of a closed chamber containing six tubes were charged with the red-fungus infusion and three with the black, while a second chamber of three tubes was charged with the yellow-fungus infusion. They were all boiled for five minutes after their intro- duction into the chambers. For two or three days all the infusions continued clear; but they subsequently broke down, every tube of the nine becoming-turbid with organisms and covered with scum. Examined microscopically on the 8th of November the red-fungus infusion was found charged with a mul- titude of spore-like bodies, massed in some places con- tinuously together, in others floating freely in the liquid. Among these ran long filaments, dotted with spore-like specks from beginning to end. There was a consider- able number of Vibrios in one of the tubes. The black- fungus infusion contained a mixed population of Vibrios and Bacteria with spore-filled filaments. Swarms of Bacteria were observed in the red-fungus infusion. Suspicious of the chambers in which these infusions had been exposed, I had three new ones constructed and provided with new tubes. A fresh supply of fungi was sent to me from Heathfield, a tree fungus being, VITALITY OF PUTREFACTIVE ORGANISMS. 159 however, substituted for the black one used in the former experiments. On the 1st of November the three infusions were very carefully introduced into three chambers, a chamber being devoted to each in- fusion. I thought it advisable to vary the period of subsequent boiling. One tube of the yellow fungus was therefore boiled for five, one for ten, and one for fifteen minutes; but as it was difficult to save the infusion from waste when the boiling was long continued, one tube of each of the other two infusions was boiled for five minutes, and the other two for ten. Tubes charged with the respective infusions were exposed at the same time to the common air. In two days the outside tubes containing the red- and yellow-fungus infusion became turbid and covered with the fatty scum so prevalent in our laboratory this year. No scum had formed on the surface of the ex- posed tree-fungus infusion, which, to casual observation, appeared quite black. Closer scrutiny, however, showed that it transmitted the deepest red of the spectrum, and was apparently quite free from floating matter. It changed rapidly during the night of the 3rd, and on the morning of the 4th of November the bottom of this tube was found laden with a heavy dark-brown precipi tate, while numerous dark-brown flocculi floated in the liquid overhead, which had become almost as clear and colourless as water. Under the microscope the dark- brown mass resolved itself into confused moss-like patches and long cylindrical sheaths dotted throughout with small dark specks. These filaments with spore- like specks have been of very frequent occurrence in this inquiry. The deportment. of the closed chambers was as fol- lows:— 1. Yellow fungus: the liquid in the three tubes remained perfectly and permanently clear and without 160 THE FLOATING-MATTER OF THE AIR, a trace of the scum which loaded the infusion outside. 2. Red fungus: one of the three tubes became thickly turbid, while the two others maintained their pristine brilliancy. 3. Tree fungus: one of the tubes became thickly turbid, the two others remained permanently clear. Iasked myself why should one tube of the red-fungus give way and the others remain intact? The answer seemed at hand. The turbid tube had been boiled for only five minutes, while the clear ones had been boiled for ten. On consulting the adjacent chamber this pos- sible explanation was blown to the winds, for here the turbid tube had been boiled for ten minutes, while its untainted neighbour had been boiled for only five. Thus, although the more careful repetition of the experiments did not secure every tube from infection, the escape of seven out of nine tubes entirely destroys the presumption of spontaneous life-development which the first experiments might suggest to some minds. Wishing to observe more attentively the action of common uncleansed air upon boiled fungus-infusions, a tray of 100 tubes was charged with them on the 14th of October. Thirty-five tubes were filled with black, thirty-five with yellow, and thirty with red-fungus in- fusion. On the 16th of October every one of the yellow- fungus tubes was turbid and covered with a thick, cohe- rent, cobweb-like scum. The surfaces of the black- fungus tubes were also sprinkled with spots of white scum. Turbidity was the only change observed in the red-fungus tubes. They were wholly free from scum. Examined microscopically on the 2nd of November the yellow-fungus tubes were for the most part found swarming with exceedingly small and active Bacteria; the red-fungus tubes also swarmed with Bacteria, some beaded Vibrios being mingled with them. In many of the tubes examined galloping monads appeared, attain- VITALITY OF PUTREFACTIVE ORGANISMS. 161 ing an astounding development in the black-fungus infu- sion. Patches of moss-like matter would appear here and there in the field of the microscope; and it was no uncommon thing to see from ten to twenty monads nestling and quivering in this ‘moss,’ and darting actively in and out of it. They put me in mind of frogs amid their spawn; and as I looked at them my belief in the animality of the one was almost as strong as in that of the other. Almost every patch of spawn- like matter had its colony. In some cases hardly any thing but monadz was to be seen; but in others the crowding of active Vibrios was so great that the monads wholly retreated from the field. § 10. Infusions of Cucumber, Beetroot, &e. The fungi having disappeared on the approach of winter, I turned to cucumber and beetroot, not expecting that their sterilization would offer any difficulty. Two closed chambers were accordingly prepared, left for the proper time in quietness, and on the 7th of November were charged, the one with the cucumber- and the other with the beet-root infusion. In a few days the infu- sions in both chambers broke down, first losing their transparency and afterwards loading themselves with fatty scum. Thus perplexities accumulated. On the 18th of November twenty-four Cohn’s tubes! were charged with infusions of cucumber, beetroot, par- snep, and turnip, six tubes being devoted to each infu- sion. They were placed in a vessel of cold water, raised ‘gradually to the boiling-point, and maintained at the builing temperature for ten minutes. Before their re- moval from the hot liquid they were one and all plugged with cotton-wool. 1 See § 4. 162 THE FLOATING-MATTER OF THE AIR. On the 30th of November all the infusions were thickly turbid throughout and heavily coated with scum. From some of the precautions already mentioned it may be inferred that before this point of the inquiry had been reached, I had begun to suspect the atmo- sphere in which I worked. Hay of various kinds, both old and new, had been exposed and shaken about in the laboratory, the air of which doubtless contained multi- tudes of spores which diffused and insinuated themselves everywhere. So, at all events, I reasoned. On the 20th of November, therefore, I had infusions of cucum- ber, beetroot, parsnep, and turnip prepared, far from the laboratory, in one of the highest rooms of the Royal Institution, and introduced into four new chambers of three tubes each. JI deemed the precaution of prepar- ing the infusions and introducing them in the distant room sufficient. Accordingly, when the chambers were charged they were carried down, and the infusions boiled in the laboratory. Two days afterwards the parsnep alone remained clear This, however, was only a respite, for a day or two sub sequently it fell into the condition of its neighbours. On the 30th of November both turnip- and parsnep- infusions were turbid throughont, and laden at the surface with thick fatty scum. The cucumber was also heavily laden with scum, which sent long streamer-like filaments into the subjacent liquid. The beetroot agreed with the others in becoming turbid, but differed from them in remaining free fromscum. In no case last year did turnip-infusion show the deportment here described. Knowing, then, from multiplied experiments, that turnip possessed no inherent power of life-development, the conclusion was irresistible that its present behaviour, and with it the behaviour of cucumber, beetroot, and parsnep, were due to infection from without. VITALITY OF PUTREFACTIVE ORGANISMS. 163 I once more tried removal to a distant room, with the added precaution of not only introducing the infu- sions into the chambers upstairs, but of boiling them there. It had been noticed that when the test-tubes were withdrawn from the oil-bath, and the discharge of steam into the chambers ceased, a somewhat violent entrance of the air into the cooling chamber was the consequence. To sift such air of its germs, both the funnel of the pipette and the open ends of the bent tubes were carefully stopped with cotton-wool. The wool was never removed from the funnel, and it was not removed from the bent tubes until the chamber had thoroughly cooled. The same vegetables were operated on, viz. cucumber, beetroot, turnip, and parsnep. On the 25th of November four chambers were charged with the infusions. On the 30th they were one and all covered with a layer of deeply pitted and corrugated fatty scum. Thus far, then, I was defeated in my efforts to escape contamination. During these experiments a fact was observed which repeated itself afterwards in other instances. Samples of the different infusions were always exposed to the common air beside their respective chambers, and in general these outside samples became turbid and covered with scum a day or so before the interior tubes gave notice of breaking down; but here, in the case of the turnip, the outside tube continued pellucid and free from life for some time after the inside ones had become turbid with organisms. How could this be? The case of my two trays placed one above the other last year! suggested itself to my memory. In regard to life-de- velopment it was then found that the lower tray was always in advance of the upper one. As pointed out 1 Phil, Trans., vol. clxvi. p. 68. 164 THE FLOATING-MATTER OF THE AIR. at the time, the absence of agitation which permitted the germs to sink into its tubes was the cause of the quicker contamination of the lower tray. No other cause appeared to me assignable in the present instance. By some means or other germs had insinuated them- selves into my closed chamber, where the tranquillity of the air permitted them to sink into the infusion, and thus produce effects in advance of those produced by the unquiet air outside. So, at all events, I reasoned. But how could the germs get into the chamber? I could, at the moment, fix only upon one way. The weather had changed from warm to cold and from cold to warm. This genial outside temperature sometimes caused the air surrounding the infusions to rise to up- wards of 90° Fahr., and we had often to work in this heat. To moderate it, I sometimes partially turned off the gas, thus lowering the temperature of the room 10° or more. The contraction of the air within the closed chambers followed as a matter of course, and the bent tubes being open, I thought the entrance of the external air might be sufficiently rapid to carry germs along with it. A new chamber of six tubes was therefore prepared upstairs, three of its tubes being charged with cucum- ber- and three with turnip-infusion on the 27th of November. The pipette funnel and the bent tubes were plugged above with cotton-wool, which was not removed from them afterwards. I took care, moreover, not to alter the gas-stoves in any way. My care was nugatory. In three days every tube of the six was laden with life. Another chamber of six tubes, charged on the 30th of November with cucumber-infusion, and two additional ones prepared on December 1st, shared the same fate. Slices of cucumber were next digested for three VITALITY OF PUTREFACTIVE ORGANISMS. 165 hours ; the infusion was filtered, boiled, and such pre- cipitated matter as appeared on boiling was removed by refiltering. The liquid thus prepared was introduced into five thick glass tubes, which were hermetically sealed, placed in a cold oil-bath, gradually heated to 230°, and maintained at that temperature for a quarter of an hour. The tubes being removed and permitted to cool, the infusion was introduced into a chamber of six tubes, and boiled there for five minutes. The previous superheating of the infusion did not even retard the development of life, for in less than two days every tube in the chamber swarmed with Bacteria. Thus far, then, every attempt ata solution was unsuc- cessful. But why, it may be asked, attempt such solutions ? Was it not mere prejudice against the doctrine of spon- taneous generation that prevented me from frankly submitting to the apparent logic of facts, and admitting the experiments just recorded to be a demonstration of the doctrine? By no means. The only prejudice I feel is the wholesome repugnance to accepting momen- tous conclusions on insufficient grounds. Hume's cele- brated argument has its application here. Taking an- tecedent experience fully into account, it was far easier for me to believe my knowledge imperfect, or my present work erroneous, than to believe the doctrine of spontaneous generation true. § 11. New Experiments on Animal Infusions. Contradictory results. In the course of this inquiry I was continually re- minded of my experiments in 1875, when the most complete immunity from Bacterial or fungoid life was so readily secured. I had operated many times with 166 THE FLOATING-MATTER OF THE AIR. turnip, never finding the least difficulty as to its steriliza- tion. It is certain that the care bestowed in preparing the turnip-infusion on the 20th of November, 1876, was greater than that bestowed upon the same infusion in 1875. But whereas the latter was invariably sterilized by five minutes’ boiling, remaining afterwards as pel- lucid as distilled water, the former, three days after its preparation, became thickly turbid and swarming with life. I extended the present inquiry to other substances whose deportment was familiar to me last year, some of whose infusions, indeed, still remain with me as clear as they were on the day of their preparation. On the Ist of December. for example, infusions of beef, mutton, pork, herring, haddock, and sole were prepared, and introduced into six closed chambers, each containing three tubes. On the 5th of December the pork, beef, mutton, and haddock were all covered with a fatty corrugated scum. A second chamber, containing artichoke-infusion, prepared at the same time, was found on the 5th more turbid than any of the animal infu- sions, and equally covered with scum. In the animal infusions, indeed, the body of the liquid underneath the scum maintained a surprising brilliancy, the develop- ment of life being confined to the layer in immediate contact with the atmospheric oxygen. On the 5th of December the herring- and sole-infu- sions were both clear; but this was only a respite, for on the 6th white spots appeared on the latter, which extended until they covered the whole surface. The herring-infusion remained clear for a week, after which small specks began to appear on its surface. They never reached the development of the scum which coated the other infusions. It sometimes occurred to me that the oil of this fish exercises a certain antiseptic action. - VITALITY OF PUTREFACTIVE ORGANISMS. 167 Last year I preserved infusion of herring perfectly pellucid for months, even in a chamber so leaky that the light could be seen through its chinks. I had, moreover, no failure with any of the animal infusions here enumerated. Last year they all remained sweet and clear; this year, with far greater precaution, I failed to protect any of them from putrefaction. Re- flection on these results, renders, I think, but one con- clusion possible to the scientific mind. It will be loth to assume that mutton, beef, pork, haddock, herring, and sole had totally changed their natures, and con- tracted qualities and powers in 1876 which they did not possess in 1875. But if the origination of the observed life be denied to the infusions themselves, there is but one other source to which it could be re- ferred, namely, atmospheric contamination. It became, indeed, more and more obvious to me that, in consequence of increased vitality or virulence in the contagia afloat this year, liberties in the preparation of the infusions or defects in the construction of closed chambers which would have been of no moment a year ago were sufficient to ruin the experiments, and render nugatory the usual means of sterilization. Against such defects I continued to struggle. With a view to stopping all chinks and crannies which might permit of the entrance of contamination, I had some of the chambers carefully coated with oil-silk and others covered with three coatings of strong paint; and as failure had attended my efforts to procure an uninfected atmosphere upstairs, I had the entire apparatus used for digesting, filtering, and boiling removed to a store~ room at the base of the Royal Institution. The floor of the room was of stone, and it was covered by no carpet. Prior to going into it, moreover, I caused my assistant to remove the clothes which he had previously 168 THE FLOATING-MATTER OF THE AIR. worn in the laboratory and to dress himself in others. The infusions prepared under these conditions were cucumber, melon, turnip, and artichoke, which, from beginning te end, were operated on below stairs. Two chambers were devoted to each infusion, and after the usual boiling in the chambers they were permitted to remain in the store-room throughout the night, being transferred to the warm laboratory next morning. I fully expected that the majority of these chambers would prove sterile. I did not expect to find them all in this condition, because the chambers had been put together in the laboratory, the air of which must have deposited its germs not only on the glycerine-coated interior of the chambers, but also on the inner surfaces of the test-tubes. My expectation, moderate as it was, was not realized. The only noticeable peculiarity in the deportment of the infusions was that they yielded tardily, but in the end every one of: them, without exception, broke down. Was the infection in this case aaa from the air of the store-room? I think not; and for this reason :— On the 27th of December four hermetically-sealed flasks, charged with a cucumber-infusion which had remained perfectly pellucid for some weeks, were opened in the store-room; four similar flasks, charged with the same infusion, were opened at the same time in the laboratory. On the 31st of December the whole group of the latter four was found invaded by organisms, while those opened in the store-room contracted no infection and developed no life. Ido not imagine, therefore, that the air of the store-room had anything to do with the con- tamination of the infusions contained in the closed cham- bers, but conclude that the contagium already existed in the chambers when they were taken down stairs. They acted as infected houses placed in salubrious air. VITALITY OF PUTREFACTIVE ORGANISMS, 169 § 12. Infusions protected by Glass Shades containing Calcined Air. I have already described this mode of experiment.! The shades stood upon circular plates of wood, each supported on a tripod (see fig. 16). Under each shade were two upright rods of stout copper wire, and stretch- ing from rod to rod was a spiral (p) of platinum wire. The copper wires passed through the slab of wood, . their free ends being in the air. The rim of each shade was surrounded bya collar of tin attached by wax to the slab, with a space of about half an inch between the collar and the glass. After the introduction of the infu- sions and the mounting of the shades, this annular space was packed with cotton-wool. The aim here was to destroy the floating matter of the air by the incandescent pla- tinum spiral. The air heated by the spiral would of course expand, passing outwards through the cotton- wool, while the air re-entering, on the cooling of the shade, would be duly sifted by the wool. In my former experiments five minutes’ incandescence sufficed to render the air absolutely inoperative on infusions exposed to its action. © In the present experiments the period of incan- } Phil, Trans. vol. clxvi. p. 50. 170 THE FLOATING-MATTER OF THE AIR. descence was doubled, ten minutes being allowed in- stead of five, while the wire was raised to the highest possible degree of incandescence. The infusions em- ployed were turnip and cucumber, a group of three tubes being charged with each. After the air had been calcined, the infusions were boiled for five minutes in an oil-bath. With this mode of treatment not a single failure occurred in 1875, turnip-infusion being among the number of liquids thus treated. This year two days sufficed to render every one of the six tubes turbid with organisms and to cover the infusion with a heavy scum. I, however, had occasion to doubt the closeness of these shades. The wax intended to seal the junction of the tin collar with the plate of wood had cracked and yielded here and there, and the entry of contamination through such cracks was possible. - Six new shades were therefore mounted and surrounded by collars which were imbedded in white lead and firmly screwed down to the plate of wood. The height of the collar, which measured the depth of the filtering layer of wool, was much greater than it had ever been last year. As before, the period of incandescence was ten minutes, during which the platinum spiral was brought as close as possible to its point of fusion. Each of these six shades covered a group of three test-tubes. Two such groups were charged with turnip, two with cucumber, and two with artichoke-infusion. The infusions, as usual, were boiled for five minutes after calcination. They were all brilliant when pre- pared; but in two days every one of them had become turbid, and had covered itself with a fatty scum. This gradually augmented until it reached in some of the tubes a thickness of half an inch. The weight of the scum caused it in some cases to bag downwards, form- ing a kind of inverted cone, the apex of which was VITALITY OF PUTREFACTIVE ORGANISMS. 171 more than an inch from its base. These bags finally broke and scattered their organisms in the subjacent liquid. Thus the conflict went on. § 13. Further precautions against Infection. At the beginning of December, my attention being keenly aroused by those successive failures, I watched more closely than I had previously done the filling of the test-tubes through the pipette. Now and then I noticed minute bubbles of air carried down with the descending infusion. On escaping from the end of the pipette, these small bubbles, I concluded, would break, and scatter such germs as they contained in the air of the chamber. Last year I should have found it difficult to believe that a cause so small could lie at the root of the observed anomalies; but this year I had learned to respect small causes, and accordingly took measures to effectually exclude the air. On December 4th three chambers, which had been previously left quiet for several days, were charged with carefully prepared cucumber-infusion, and two other chambers with turnip-infusion prepared with equal care. The following precautions were taken :—The funnel of the pipette formerly employed was broken off from its shank, and for it was substituted a ‘ separa- tion-funnel’ with a glass stopcock. This was connected by closely fitting india-rubber tubing with the shank of the pipette. But before the connexion was made, the funnel was filled with the infusion, and the stopcock turned on for a moment, until the liquid issued from the orifice below. The stopcock being then turned off, the flow of the liquid ceased, and the column in the shank below the stopcock was supported by atmospheric pressure. A pinchcock nipped the india-rubber tube at 172 THE FLOATING-MATTER OF THE AIR. its centre. The portion of tubing above the pinchcock was then filled with the infusion, and the end of the separation-funnel introduced into the tube, all air being thus excluded. On turning on the stopcock and releasing the pinchcock, the liquid passed slowly down the shank of the pipette, filling it wholly. The point of the shank was then placed in succession over the test-tubes, the infusion entering them witbout a single associated bubble. The arrangement was not absolutely perfect, but it was an improvement upon previous ones. As before, the charged chambers were placed in a room the air of which was maintained at a temperature of about 90° Fahr. The result was as follows :—Of the two turnip cham- bers prepared as just described, on the 4th, one had completely given way on the 6th. In the other cham- ber two out of the three tubes had given way, but the third remained permanently brilliant. Previous to this series of experiments I had never succeeded in saving even a single tube of cucumber-infusion ; here, how- ever, two out of the three chambers charged with it remained perfectly clear for many days. Subsequently one of these chambers yielded in part, through an accident, but the other chamber is as brilliant at this moment as it was on the day of its preparation many months ago. Now, as regards inherent power to generate life, the infusion of this chamber was in precisely the same condition as its two neighbours. They, one and all, contained the same infusion; and there is no way of accounting for the observed difference of deportment save by reference to contamination from without. Here we are certainly on the trace of the enemy which has given us so much trouble. On the 5th of December two additional cases were VITALITY OF PUTREFACTIVE ORGANISMS. 173 charged with infusion of melon prepared in the usual way; and on the 12th of December I subjected both these chambers, and those prepared upon the 4th, to a very close scrutiny. The result was instructive. After the introduction of the infusions, and prior to the re- moval of the separation-funnel, the india-rubber tubing connecting the latter with the shank of the pipette was perfectly closed by the pinchcock. Provided the clasping of the india-rubber tube round the shank of the pipette were perfectly air-tight, the liquid contained in the shank ought to remain there, supported by atmospheric pressure. If, however, the india-rubber tube failed to clasp with sufficient tightness the pipette- shank, air would insinuate itself between the two, and the depression of the liquid would be the consequence. The result, observed upon the 12th was this:—In two only of the seven chambers prepared on the 4th and 5th was the liquid column found perfectly supported ; and only in these two chambers were the test-tubes, which contained cucumber-infusion, without exception pellucid. In the five remaining chambers the liquid columns, which had completely filled the pipette-shanks on the 4th and 5th, were found more or less depressed. The tubes in one of the chambers, containing melon-infusion, had become rapidly turbid and covered with scum. The pipette-shank in this case was found entirely emptied of its liquid and filled with air. Another chamber had nine inches, while a third had seven inches of its pipette-shank filled with air. In a fourth chamber only one inch of the pipette-shank was filled with air; here one out of a total of three tubes remained pellucid. Thus, where the closure above was perfect, we had in this instance perfectly pellucid infusions ; where it was grossly defective, the infusions gave way in all the tubes ; 174 THE FLOATING-MATTER OF THE AIR. while where the closure was but slightly defective we had the escape of a fraction. The defects thus revealed came, I concluded, into play when the infusions were introduced, the descending column of liquid sucking in minute air-bubbles between the india-rubber tubing and the pipette, thus carrying with it the external contagium. Few seem aware of the precautions which are sometimes essential to save the experimenter from error in inquiries of this nature. Even with some of our best and most celebrated observers I find no adequate sense of the danger involved in their modes of experimentation. § 14. Eaperiments in the Royal Gardens, Kew. But it was only in exceptional instances, dependent on the state of the air, that even precautions such as those described in the foregoing section secured freedom from contamination. The contagium seemed omni- present and persistent, and whether it was local or general—due to the accidental condition of our labora- tory, or to an epidemic of the air—became a question with me, not by any means to be decided offhand. On this point, then, I held judgment in suspense. The in- fection was, to all appearance, fully accounted for by reference to the conditions under which I worked; but as regards outbreaks of epidemics the autumn had been a remarkable one, and it seemed well worth investigating whether it was not also a period prolific generally in the germs of putrefaction. I resolved therefore to break away wholly from the Royal Institution, and, thanks to the friendly permission of Sir Joseph Hooker, I was enabled to transfer my apparatus to Kew Gardens. By the enlightened mu- VITALITY OF PUTREFACTIVE ORGANISMS. 175 nificence of Mr. Jodrell, a new and very complete labo- ratory had been just erected there, and in it I sought a purer air than I could find at home. My chambers hitherto had been constructed of wood, but those to be tested at Kew were made of block-tin, and they were carried direct from the tinman’s to the gardens without being permitted to come near the in- fected air of Albemarle Street. At Kew the test-tubes - employed were first cleansed with carbolic acid, then washed with a solution of caustic potash, afterwards swept out with distilled water, and finally raised almost lo the temperature of redness bya Bunsen-flame. They were then fitted air-tight into the chambers with white- lead and tow. The chambers were closed on the 3rd of January, and allowed to remain quiet until the 8th, when the two most refractory liquids that I had encountered in the laboratory of the Royal Institution were introduced into them. These were infusions of cucumber and melon. There were two chambers devoted to each infusion—four in all; and each chamber embraced three large test-tubes. The period of boiling was that found effectual last year, i.e. five minutes. The temperature of the room in which the chambers were placed was maintained, partly by hot-water pipes and partly by a gas-stove, at about 90° Fahr.—-a temperature which had been proved eminently favourable to the development of Bacteria. Tubes containing the same infusions were at the same time exposed to the common air of the Jodrell labora- tory. These became rapidly turbid and covered with scum. My interest and anxiety during the early days of the trial of the protected tubes may be imagined. After eleven days’ exposure they showed no signs of giving way. On the 19th of January the four chambers 9 176 THE FLOATING-MATTER OF THE AIR. were removed ina van from Kew, and shown in the evening of that day to the members of the Royal Institution, including many eminent Fellows of the Royal Society. The infusions were one and all brilliant, no trace either of turbidity or scum being found associ- ated with any of them. During all my previous efforts (and they had been very numerous) I had never suc- ceeded in saving a single tube of melon-infusion ; here, however, every tube of both chambers was intact. The epidemic was thus localized, the obvious cause of it being the contaminated air of our laboratory. _ A couple of days subsequent to the removal of the chambers from Kew, a single tube of the cucumber- infusion became turbid, its two neighbours in the same chamber remaining intact. Not one of the other tubes, either of melon or cucumber, gave way. They all remained as pellucid in London as they had been at Kew. Their removal from Albemarle Street to the city last year ruined many of our sterilized chambers. I was not therefore prepared to see so little damage done by the transport from Kew, It may be remarked, in passing, that this infection of an infusion by mere mechanical shaking is an obvious proof that the contagium is not a gas or vapour, but that it consists of particles capable of being detached from the interior surface of the chamber, and endowed with the power of passing into active life. Two other chambers were exposed at the same time in the Jodrell laboratory, the one containing beef- and the other sole-infusion. They are by no means so sensitive as the cucumber and melon, still one of the three beef-tubes broke down, becoming thickly turbid throughout. Right and left of this tube its two com- panions remained perfectly transparent. Asan illustra- tion of the externality of the contagium, the result was VITALITY OF PUTREFACTIVE ORGANISMS. 177 more conclusive than it would have been had all three tubes remained intact; for had the power of developing the organisms which produced the turbidity been in- herent in the infusions, its action would not have been confined to a single tube. It will be understood that when the chamber is lifted from the oil-bath in which its infusions are boiled, the air within the chamber contracts, and an indraught is the consequence. If the entering air be properly sifted, by passing it through cotton-wool plugs, no harm is done; but if it enter an aperture unsifted, it carries its motes along with it. In the beef-chamber just re- ferred to an aperture of this kind, about the size of a pin-hole, was detected. This obviously was the door through which the contagium entered. Through a similar but graver defect in its chamber the sole- infusion also broke down; but in a subsequent experi- ment with sole-infusion in the Jodrell laboratory, two- thirds of the whole number of tubes charged with it remained free from all trace of life. § 15. Haperiments on the Roof of the Royal Institution. With a view to making, nearer home, experiments similar to those made at Kew, I had a wooden shed erected on the roof of our laboratory. The shed was provided with benches, water and gas-pipes, and a stove for heating. To an infusion of cucumber, which I had found extremely intractable in the laboratory, my atten- tion was first directed. Two tin chambers of three tubes each were prepared, and transferred to the shed fiom the workshop where they were made, without 178 THE FLOATING-MATTER OF THE AIR. being permitted to enter our laboratory. The cucumber used for the infusion was also kept clear of the infected air; it was sliced and digested in the shed, the infusion was there filtered, introduced into the tin chambers, and boiled subsequently for five minutes. The result was not that expected. Not a single tube of either of these two chambers escaped con- tamination. They one and all behaved like the same infusion in the infected laboratory, becoming in three days turbid throughout and laden with fatty scum. I have been daily and hourly impressed with the parallelism between these phenomena of putrefaction and those of infectious disease. A further illustration of this parallelism is here presented to us. The clothes of my assistants who prepared the infusion in the shed had been worn in the laboratory, a transfer of infection by one of the modes of transport known to every physician being the result. The thoughtful physician cannot indeed fail to see the absolute identity of deport- ment between the contagia with which he is familiar, and those assailants of my infusions against which I have been contending so long. With regard to the shed my first step, after this preliminary failure, was to disinfect it. This was done by washing every part of it, first with a mixture of carbolic acid and water, and secondly with a solution of caustic potash. When the whole was well dried, new tin chambers furnished with new tubes were introduced. Cucumbers and beef fresh from the market were also digested in the shed, my assistant taking care to cover his legs with clean linen trowsers, and his body with a new blouse. There was one chamber devoted to the cucumber and another to the beef. Into the former the infusion was introduced on the 19th, and into the VITALITY OF PUTREFACTIVE ORGANISMS. 179 latter on the 20th of March; each infusion was boiled for five minutes after its introduction. Let us compare results and draw conclusions. Ata distance of eight yards from the shed, viz. in the labora- tory, infusions both of beef and cucumber refused to be sterilized by three hours’ boiling. Indeed I have samples of both infusions which have borne five hours’ boiling and developed multitudinous life afterwards. But the upshot of this experiment in the disinfected shed is, that every tube of the two chambers, though boiled for only five minutes, contains an infusion which, at the present hour, is as limpid as the puvest distilled water. What shall we say, then? is the infusion in the laboratory endowed with a generative force denied to the same infusion in the shed? Irrespective of the condition of the air, can a linear space of eight yards produce so remarkable a difference? It is only the confusion of mind still prevalent in relation to this sub- ject that renders such a question necessary. Let me add that it suffices simply to wave a bunch of hay in the air of the shed to make it as infective as the labora- tory air. Even the unprotected head of my assistant when his body was carefully covered sufficed in some cases to carry the infection. If anything were needed to illustrate the extra- ordinary care necessary on the part of physicians and surgeons, both as regards the clothes they wear and the instruments they use, such illustrations are copiously furnished by the facts brought to light in this inquiry. 180 THE FLOATING-MATTER OF THE AIR. § 16. Preliminary Experiments on the Resistance~ limit of Germs to the temperature of Boiling Water. While continuing the conflict and experiencing the defeats recorded in the foregoing pages, a remark of Professor Lister’s sometimes occurred to me. To apply the antiseptic treatment with success, the surgeon must, he holds, be interpenetrated with the conviction that the germ theory of putrefaction is true. He must not permit occasional failures to produce scepticism, but, on the contrary, must probe his failures, in the belief that his manipulation, and not the germ-theory, is at fault. This may look like operating under a prejudice; but Professor Lister’s maxim is nevertheless consistent with sound philosophy and good sense; and if I permitted a bias to influence me in this inquiry, it was one fairly founded on antecedent knowledge, which led me to conclude that the long line of failures above referred to would eventually be traced to my ignorance of the conditions wherehy perfect freedom from contamination was to be secured. I laboured to discover these conditions, and to learn something more regarding the nature of the contami- nation—its origin, persistence, and manner of action. When these researches began, five minutes’ boiling, as I have frequently stated, sufficed to sterilize the most diversified infusions. Here we have frequently extended the time of boiling to ten and fifteen minutes, and, in some cases glanced at above, to immensely longer periods, without producing this result. I desired more exact knowledge as to the limit of endurance, and with this view, on the 22nd of December, had six ‘ pipette bulbs’ charged with an infusion of cucumber, sp. gr. 1004. They were then plugged with cotton-wool, hermetically sealed, and subjected to the boiling temperature for 10 VITALITY OF PUTREFACTIVE ORGANISMS. 181 minutes. Six other bulbs, charged with the same in- fusion and treated in the same way, were boiled for 30 minutes. Finally eight bulbs, similarly charged, were boiled for 120 minutes. On the 23rd of December three of the first group of bulbs, three of the second, and five of the third, had their sealed ends filed off, and were afterwards exposed to a tolerably constant temperature of about 90° Fahr. Not one of these twenty bulbs preserved itself free from life. On the 25th of December every one of them had given way to cloudiness and turbidity. There was, however, a marked difference between the sealed and the unsealed bulbs. To the latter, it will be remembered, the air had access through the plug of cotton-wool, while to the former no air had access, save the small quantity imprisoned above the infusion when the necks of the bulbs were sealed. The aerated bulbs grew rapidly and thickly turbid, while a passing cloudiness was all that showed itself in the sealed ones. This soon disappeared, and left the in- fusions apparently intact. In fact it required some attention to detect the appearance of this fugitive life, which existed only so long as there was oxygen to sus- tain it. I have ranged the sealed and unsealed tubes side by side in groups. To the most cursory observa- tion the difference between them is obvious. The ex- periment strikingly illustrates the dependence of the special organisms here implicated on the oxygen of the air. The experiments were pushed still further on the 28th of December. Two bulbs of cucumber, two of melon, two of turnip, and two of artichoke were then plugged, sealed, and maintained at the boiling tem- perature for four hours. Six of the eight bulbs burst in the operation, but two of them, a bulb of melon and 182 THE FLOATING-MATTER OF THE AIR. one of cucumber, bore the ordeal uninjured. After cooling, their sealed ends being broken off, they were placed in the warm room. The melon remained per- manently sterile, but in two days the cucumber-infusion became turbid and laden with fatty scum. Eight similar bulbs were boiled on the same day for five hours and a half. Four of them burst, but four remained intact. Of these, two contained cucumber-, one melon-, and one turnip-infusion. Three out of the four bulbs were sterilized by the long-contibued boiling, but one cucumber-bulb passed through the ordeal unscathed. Two days after the operation it swarmed with life, and was covered with a fatty scum formed of matted Bacteria. Many similar experiments were subsequently made. On the 27th of January, for example, six bulbs of turnip infusion were boiled for 220 minutes, six for 300 minutes, and two for 305 minutes. Suspended in the air above each infusion was a sprig of old Colchester hay, this being purposely introduced to augment the chance of infection. Notwithstanding its presence the bulbs were one and all permanently sterilized. The specific gravity of the infusion was in all cases 1007. The sprigs of old hay were afterwards shaken into the liquid, but they produced no effect. For weeks afterwards the infusion remained clear. Was this im- potence to generate life due to the fact that the nutri- tive power of the infusion had been destroyed by the ‘blighting influence of heat’? Not so; for when the same infusion was infected by a sprig of fresh hay, by a small pellet of cotton-wool rubbed against the dusty shelves of the warm room, or by a speck of another infusion containing Bacteria, it never failed to develop life. The only observed difference between the effect produced by the dry hay or dust and the living Bacteria VITALITY OF PUTREFACTIVE ORGANISMS. 183 was purely a difference of time. Inoculation with the finished organisms acted more rapidly than infection with the dust, but the effects were the same in the end. On the 27th of January also nine melon-bulbs were treated exactly like the turnip, being furnished with sprigs of old Colchester hay, plugged with cotton-wool, and hermetically sealed above the plugs. Six of them were boiled for 215 minutes, and three for 220 minutes. They were one and all permanently sterilized ; but, like the turnip, all of them were open to infection by fresh hay, dry dust, or living Bacteria. The specific gravity of the melon-infusion was 1008. § 17. Further Experiments on the Resistance-limit of Germs to the Boiling Temperature. The amount of boiling which turnip-infusion failed to withstand is shown by some of the foregoing experi- ments; but to determine the limit of its resistance we must begin with shorter periods. On the 1st of March, therefore, eight groups of pipette-bulbs were charged with turnip-infusion which had been prepared in an atmosphere purposely infected with the germs of old Heathfield hay. In every case, moreover, a sprig of the same hay was placed in the air above the infusion. The bulbs had their necks plugged with cotton-wool, and were hermetically sealed above the plug. In this condition the respective groups were boiled for the fol- lowing times :— 1st. group ‘ : 5 i . 15 minutes. 2nd ,, cs @ @ «© «6 180 34 8rd, cS ee wR, TRB ae 4th ,, . . . 6 , bth ,, 75 oy 6th ,, ; 90 ~«, 7th =, ' 105, Sth " 120 Si 184 THE FLOATING-MATTER OF THE AIR. After boiling they were removed, permitted to cool, had their necks broken off by a file, and were afterwards exposed to the temperature of our warm room. It may be remarked that the infusion gradually deepened in colour from the 15-minute period, where the colouring was hardly sensible, to the 2-hour period, where the colouring became deep yellow. The effect was doubtless due to the oxidation of the infusion, which, notwithstanding the colour, was in all cases highly transparent. Two days after their preparation, every tube of the series had become turbid and had begun to cover itself with scum. On March the 6th the periods of boiling were pro- longed with a fresh infusion. Two groups of tubes were, on that day, exposed to boiling water for the fol- lowing times :— Ist group é : 5 . - 180 minutes. 2nd ,, ge Sage cael. “hg Sd hg On the 8th of March all the members of the first group were turbid and covered with scum. The second group was completely sterilized. This latter result is quite in accordance with the experiments made on the 27th of January. Turnip-infusion was then boiled for periods varying from 220 minutes to 305 minutes, complete sterilization being in all cases the consequence. These results were subsequently checked by a continuous series of experiments extending over periods of boiling varying from one to six hours. Up to three hours the infusion resisted sterilization ; but when the periods of boiling were prolonged to four, five, and six hours respectively, all the bulbs became permanently barren. The liquid continued in the highest degree transparent, and in colour a brilliant orange-brown. VITALITY OF PUTREFACTIVE ORGANISMS. 185 Experiments intended to determine the limit of resistance of cucumber-infusion were made on the 24th of February. Nine pipette-bulbs were then charged, plugged, hermetically sealed, and subjected to the boil- ing temperature for the following times :— 1st bulb F ‘ : ‘ $ 15 minutes. 2nd ,, ‘ : : ; : 30 3 3rd, > So al Jo Abe 4th ,, ; § : ‘ ‘ 60 . bth ,, Lg oe tet fel. Geer. 6th ,, eh A cae gl Jat Ha a 7th ,, : ; : : . 240 5 8th ,, al fel a tnt Jel ees 3, oth ,, Fst. mn. Jat let eggs After boiling and cooling they had as usual their ends broken off by a file. The result here was that at the 5th bulb, which corresponded to a boiling for two hours, the life-development suddenly ceased. All the tubes boiled from three to six hours inclusive were com- pletely sterilized. The infusion in this case had been diluted by an accident, so that its specific gravity was not much above that of distilled water. On the 28th of February, therefore, a fresh infusion having a specific gravity of 1006 was prepared, and introduced into a series of bulbs exactly as in last experiment. The bulbs were exposed to the boiling temperature for the following times :— 1st bulb : ‘ é 5 A 15 minutes. Qnd ,, ‘ F i ‘ F 80 53 8rd, . ‘ : 3 . 45 - 4th ,, et 2, cts oh SOE Ge 5th ,, ‘ : g ‘ . 120 6th ,, - 3 A ; . 180 is 7th ae a ea |) ae 8th ,, ; , 3 j . 800 oth ,, ey Gh he ce REE OG, 186 THE FLOATING-MATTER OF THE AIR. The result here was that at the 6th bulb, which cor- responded to three hours’ boiling, the life-development suddenly ceased. All the bulbs boiled from 15 minutes to 180 minutes inclusive proved fruitful; while from 240 minutes to 360 inclusive all were completely sterilized. As in the case of the turnip-infusion, the cucumber sub- jected to long periods of boiling assumed an orange- brown tinge. Comparing these results with those obtained with the turnip-infusion, it will be observed that cucumber and turnip exhibit about the same resistant power: three hours’ boiling, and less, failed to sterilize both of them; four hours’ boiling, and more, rendered both of them permanently barren. The cucumber-infusions prepared on the 22nd and 28th of February were connected with the atmosphere through the cotton-wool plugs; but no attempt had been made to remove its floating matter from the air above the infusions. On the 22nd, however, four bulbs of the infusion were also prepared, charged with filtered air, left unplugged, and hermetically sealed. The same was done with four bulbs on the 28th of February. Each group was subjected to periods of boiling of 15, 30, 45, and 60 minutes respectively. All of them became turbid; but it was interesting to notice the gradual and obvious fall of life from the 15-minute to the 60-minute period. Could the Bacteria have been counted, and the result graphically represented, the ordinate corre- sponding to the abscissa 15 would have been found very considerably longer than that corresponding to the abscissa 60. The method of experiment here for the most part pur- sued was employed by Spallanzani and Needham. It was afterwards extensively applied by the late excellent VITALITY OF PUTREFACTIVE ORGANISMS. 187 Professor Wyman, of Harvard College, while in 1874 it was materially refined and improved upon by Dr. William Roberts, of Manchester. The method is ham- pered by one grave doubt. The air, plus its floating matter, is imprisoned in the sealed bulbs, so that the heat applied has not only to destroy the germs clasped by the infusion, but also those diffused through the supernatant atmosphere. Now it is not certain whether an amount of heat which would be absolutely destruc- tive to a germ embraced by a hot liquid may not be wholly ineffectual when acting on a germ floating in vapour or air. Throughout Spallanzani’s and Needham’s experiments, throughout those of Wyman and Roberts, and throughout my own, as reported in this section and the last, this possibility of error runs. Such ex~ periments, in short, do not enable us to state with certainty the temperature at which an infusion is steri- lized, because the germs which most pertinaciously oppose sterilization may not belong to the infusion at all, but to the adjacent air. The most astonishing cases of resistance to steriliza- tion observed by Wyman were associated with this particular mode of experiment. The et} possible action of the uncleansed air, moreover, was in his case augmented by the fact that he employed quantities of liquid, very small in comparison with the size of his flasks. In some of his earlier experiments the volume of air was more than thirty times that of the infusion. These relative volumes are represented in the annexed figure \_ (fig. 17), copied from Wyman’s Memoir 7 of 1862.' 1 Silliman’s American Journal, vol. xxiv. p. 80. 188 THE FLOATING-MATTER OF THE ATR. § 18. Change of Apparatus. New Experiments with Filtered Air. The source of possible error referred to in the last section had been long present to my mind, and I had already taken measures to avoid it. On the 2nd of January, 1877, an infusion of turnip (sp. gr. 1006) and an infusion of melon (sp. gr. 1008) were prepared and introduced into a series of pipette-bulbs in the follow- ing manner :—One end a, fig. 18, of a glass T-tube was connected with an air-pump, the other end 6 was closely plugged with cotton-wool, while to the third branch of the T-tube the neck of the pipette-bulb A was attached by india-rubber tubing. A piece of the same tubing, furnished with a pinchcock p, was also attached to the free end of the T-tube beyond the cotton-wool. The bulb A was exhausted three times in succession, the pinchcock p being closed, and was three times filled with filtered air, the pincheock being opened. At the third exhaustion the bulb was raised to a very high temperature by a Bunsen flame, and finally filled with filtered air. It was then plunged for a minute into ice- cold water, from which it was afterwards removed, detached from the T-tube, and then charged with the infusion by means of a narrow pipette, fe, shown at the top of B, fig. 18. The rationale of the above proceeding is this:—On quitting the ice-cold water for the warmer air of the laboratory, expansion of the air within the bulb would occur. This would cause a gentle motion from within outwards, opposing all indraught of contaminated air. The entry of the infusion into the bulb would, I thought, also promote this outward motion. On the removal of the pipette, which occupied but a very small portion of the VITALITY OF PUTREFACTIVE ORGANISMS. 189 neck of the bulb, a little warmth was applied to the latter, and during its application the neck was plugged with cotton-wool. The air entering through this plug to supply the place of the small quantity displaced by Fig. 18. the warmth would, I concluded, reach the interior of the bulb perfectly sifted of its floating matter. The necks of the bulbs were hermetically sealed, and the infusions 190 THE FLOATING-MATTER OF THE AIR. maintained for ten minutes at the temperature of boil- ing water. After a lapse of twelve hours their sealed ends were broken off by means of a file. In our experiments on the 28th of December tumnia and melon subjected to ten minutes’ boiling entirely gave way. In these present experiments, where greater care was taken, two out of the six turnip-bulbs and three out of the six melon-bulbs remained permanently barren. Even this amount of success proved afterwards so exceptional that it might be fairly regarded as accidental. On the 4th of January the experiments were con- tinued. The pipette-bulbs employed were first carefully washed with carbolic acid, which was removed as far as possible with ordinary water. They were then washed with a solution of caustic potash, and finally rinsed out with distilled water. They were not subjected to the action of the Bunsen flame. The infusions employed were turnip (sp. gr. 1006) and melon (sp. gr. 1008), eight bulbs being filled with each infusion. I could not be certain that the motion of the liquid fillet at the end e of the pipette with which the bulb s, fig. 18, had been charged, had not drawn into the neck of the bulb a modicum of the external air. In the pre- sent experiments, therefore, the method of charging the bulbs was modified in the following way:—The glass T-tube employed in our last experiments had its end a, which was to be connected with the air-pump, drawn out to a small orifice and bent as in fig. 19. The branch connected with the bulb was also drawn out to a tube of fine bore, which entered the neck of the bulb for some distance to ¢, the thicker part above being con- nected with the neck of the bulb by india-rubber tubing. The end b, as before, was plugged with cotton- wool and provided with a pinchcock, p. The object VITALITY OF PUTREFACTIVE ORGANISMS. 191 here aimed at was that the liquid should be discharged into the bulb far below the india-rubber connecting- piece, and that during the discharge it should pass only through filtered air. Fig. 19, SJ Each bulb was exhausted in the manner already described, and refilled three times in succession. When last filled it was plunged for a minute or so into 192 THE FLOATING-MATTER OF THE AIR. iced water, with the view of rendering the air within the bulb denser than that without. The pinchcock p being closed, the whole apparatus was then detached from the air-pump. On being lifted from the iced water into the warmer air there was a gentle outflow of air from a. The mode of charging the bulb was this:—The point a was well sunk into the infusion, and the associated bulb, B, was plunged into boiling water. There was an immediate outrush of air from a which bubbled through the liquid. As soon as the bubbling had relaxed a little, a being still submerged, the bulb was transferred to iced water. A shrinking of the warm air was the consequence, and through a the infusion was forced by atmospheric pressure. It descended the middle branch of the T-tube, and was discharged from its end e into the bulb. The quantity of liquid obtained by a first immersion in the iced water was not sufficient to charge the bulb; but by repeating the process of heating and chilling two or three times, the point @ never being permitted to quit the infusion, any required quantity was with ease and accuracy introduced. The neck of the bulb was finally detached from the T-piece by loosening the india-rubber tube. The bulb was then slightly warmed so as to cause an outflow of air from within, and while this outflow continued the neck was plugged with cotton-wool. It was sealed above the plug, and after the cooling of the infusion the sealed end was broken off with a file. It is not my intention to take up time in describing in detail the numerous experiments made in accordance with this method, or the variety of infusions employed in testing its efficacy. Suffice it to say that, notwithstanding all my care, the results were chequered throughout. Sometimes success VITALITY OF PUTREFACTIVE ORGANISMS. 193 would seem complete, but a repetition of the experi- ments—and I never felt safe without frequent and varied repetition—would, as before, present the success in the light of an accident. Iam, however, secure in stating that while pursuing this plan I have in some cases effected complete sterilization by an amount of boiling which, in other cases, though twenty times multiplied, has failed to produce this effect. I have, for example, placed side by side in my collection two series of organic infusions, one as pellucid as distilled water, having been rendered permanently sterile by an exposure to the boiling temperature for five and ten minutes, and a second series containing the same infusions boiled for 30, 120, and 330 minutes respectively, and which never- theless are muddy throughout and covered with scum. Even here, however, causes, other than differences of manipulation, may have contributed to the result. Weeks of labour have been devoted to these experi- ments, nor did they exhaust the trials actually made. Another mode of proceeding was this. Pipette-bulbs were prepared by having a portion of their necks drawn out to a tube of very fine bore. The open end being connected with an air-pump, the bulb was exhausted and filled with filtered air several times in succession. In the final experiment the bulb was charged with one- third of an atmosphere of cleansed air; and while this pressure was maintained by the air-pump the narrow tube was hermetically sealed. Each bulb was after- wards heated almost to redness in the flame of a Bunsen lamp. It was charged by inverting the bulb, dipping the sealed end into the infusion, and breaking it off underneath the surface. The liquid entered until the bulb was two-thirds filled, when the narrow tube was again sealed. A great number of experiments were 194 THE FLOATING-MATTER OF THE AIR. thus executed, the results of which distinctly favoured the conclusion, though they did not to my satisfaction prove it, that the resistaut germs were not to be wholly ascribed to the air, but that they had survived in the liquid. § 19. Final proof that the Resistant Germs are em- braced by the Infusion. Examples of Resistance both in Acid and Neutral Liquids. We here face a question which greatly harassed me at the time to which I now refer. It was this:— Have the germs, which under the circumstances here described produced life, been really embraced by the infusion itself during the time of heating? The liquid, it will be remembered, had to pass through the neck of the pipette-bulb, and it could not descend from the neck into the bulb without leaving a film adherent to the internal surface of the neck. This film, I reflected, might dry in part by evaporation; and it might, in doing so, leave germs behind which would be very differently circumstanced from those in the liquid. To germs thus exposed, not to the heat of water, but to the possibly less effective heat of vapour and air, the observed life might I thought be due. Before closing definitely with the proposition that the surviving germs had actually been in the liquid, the possibility to which I have just referred had to be inexorably shut out. The evil was to some extent mitigated by charging the bulb, not through its own neck, but through a narrow tube issuing at right angles to the neck. But even here a portion of the neck and of the higher interior surface of the bulb was trickled over by the infusion. The difficulty was finally met, and completely surmounted, by causing the lateral tube to issue from VITALITY OF PUTREFACTIVE ORGANISMS. 195 the centre of the bulb itself, and forcing the infusion into the bulb by atmospheric pressure, until the surface of the liquid stood clearly above the lateral orifice. To this level the liquid rose without wetting any portion of the surface against which it did not per- manently rest. The precise method pursued in preparing and charging the bulbs was this:—First, the bulb as sent to us by the glass-blower is represented at a, fig. 20. Its neck is first plugged with cotton-wool (c) and hermetic- ally sealed as at B, fig. 20. The lateral tube is then drawn out to almost capillary narrowness at o and p. The end 7 is connected with an air pump, by which the bulb is exhausted, and after two or three emptyings and fillings, it is finally charged with one-third of an atmosphere of thoroughly filtered air. While the pump attached to » maintains this pressure within the bulb, the capillary tube p is sealed with a lamp. The bulb and its appendages are then heated nearly to redness in a Bunsen flame, all life adherent to the interior surface being thus destroyed. The end p is then introduced into the infusion, pressed against the bottom of the vessel that contains it, and thus broken. The external pressure of a whole atmosphere, having but one-third of an atmosphere within the bulb to oppose it, forces the liquid through the lateral tube. It enters the bulb, gradually rising until it reaches the orifice, and rises above it. When the pressure within is exactly equal to the pressure without, two-thirds of the bulb are occupied by the liquid. The infusion then extends without breach of con- tinuity from the bulb B to the vessel in which the end p is immersed, the uncleansed air being thus completely 196 THE FLOATING-MATTER OF THE AIR. excluded. A small gas flame is carefully applied at o. The liquid within the narrow tube vaporizes, and the vapour drives the liquid to some distance right and left from the place of heating. In the absence of the liquid the fine tube reddens, fuses, and is hermetically sealed. Fie. 20. The aspect of the bulb after it has been thus charged is shown at ©, fig. 20. By this method, on the 20th of February sixteen bulbs were charged with infusions of old Heathfield hay and of a hard wiry hay from Guildford, not old. They weré divided into four groups, four bulbs in a group. VITALITY OF PUTREFACTIVE ORGANISMS. 197 Each group embraced two acids and two neutral in- fusions. They were boiled for the following times:— Ist group. tl, oS: . 10 minutes, 2nd, me cher <> rn ory SROe a, 3rd, . = 8» «+» « 80 y, 4th ,, see ee DF" After the bulbs had sufficiently cooled, their sealed ends were removed by a file. On the 21st of February, less than 24 hours after their preparation, all these bulbs showed signs of yield- ing. On the 22nd they were all turbid, while as regards the comparison of acid and neutral infusions, their condition was this:— ( Guildford neutral distinctly more turbid than Guildford acid. Scum on for- mer, none on latter. Ist group. 10 minutes.’ Old Heathfield neutral not to be dis- tinguished from old Heathfield acid. Both turbid and covered with scum. Much lightened in colour. Guildford neutral distinctly more turbid ond 20 minut than the acid liquid. and group,