GI38 
 6 
 
 
 
 
 
THE LIBRARY 
 
 OF 
 
 THE UNIVERSITY 
 OF CALIFORNIA 
 
 PRESENTED BY 
 
 PROF. CHARLES A. KOFOID AND 
 MRS. PRUDENCE W. KOFOID 
 
GASTRIC DERANGEMENTS. 
 
 HORSFORD'S ACID PHOSPHATE. 
 
 Unlike all other forms of phosphorus in combination, such as dilute 
 phosphoric acid, glacial phosphoric acid, neutral phosphate of lime, hypo- 
 phosphites, etc., the phosphates in this product are in solution, and readily 
 assimilated by the system, and it not only causes no trouble with the 
 digestive organs, but promotes in a marked degree their healthful action. 
 
 In certain forms of dyspepsia it acts as a specific. 
 
 Dr. T. G. COMSTOCK, of the Good Samaritan Hospital, St. Louis, 
 says: " For some years we have used it in a variety of derangements 
 characterized by debility, as also in chronic gastric ailments. It is ap- 
 proved of, unanimously, by the medical staff of this Hospital." 
 
 Send for descriptive circular. Physicians who wish to test it will be 
 furnished a bottle on application, without expense except express charges. 
 Prepared under the direction of Prof. E. N. HORSFORD, by the 
 
 RUMFORD CHEMICAL WORKS, Providence, R. L 
 
 Beware of Substitutes and Imitations. 
 
UBKf 
 
 COUFGf 'M 
 
 ThjE 1 
 
 Bacterial Poisons. 
 
 BY 
 
 DR. N. GAMALEIA. 
 
 TRANSLATED BY E. P. HURD, M.D., 
 
 Member of the Massachusetts Medical Society and of the Climatological 
 
 Society ; one of the Physicians to the Newburyport {Mass^ 
 
 Hospital. 
 
 GEORGE S. DAVIS, 
 
 DETROIT, MICH. 
 
 80 
 
Copyrighted by 
 
 GEORGE ?. DAVIS. 
 
 1898. 
 
TABLE OF CONTENTS. 
 
 FIRST PART. 
 CHAPTER I. 
 
 EXPERIMENTAL STUDY ON THE PUTRID POISON. 
 
 PAGE 
 
 Experiments with the Toxic Effects Produced by Sub- 
 stances Undergoing Decomposition Seybert, Gas- 
 pard, Stich Establish the Symptoms and the Lesions 
 of Experimental Septicaemia Panum Gives the 
 Proof that it is Produced by a Chemical Poison 
 Bergmann and Schmiedeberg Determine this Poi- 
 son to be an Alkaloid: Sepsine I 
 
 CHAPTER II. 
 
 THE MICROBIAL ETIOLOGY OF PUTREFACTION AND 
 OF THE INFECTIONS. 
 
 Contest between the Chemical and the Vitalist Doctrines 
 The Contradictions and the Results Obtained by 
 the Chemists Their Explanation Insufficient by 
 Reason of the Absence of the Notion of Specificity 
 Definitive Victory of the Vitalist Doctrine The In- 
 fectious Diseases are Admitted to be Caused by 
 Specific Fermentations 12 
 
 CHAPTER III. 
 THE DISCOVERY OF PTOMAINES. 
 
 Selmi Draws Attention to the Ptomaines Obtained from 
 Cadavers Leucomaines and Ptomaines of Gautier 
 The Researches of Brieger on Ptomaines 20 
 
VI 
 
 PAGE 
 CHAPTER IV. 
 
 INFECTION is AN INTOXICATION BY THE MICROBIAN 
 POISON. 
 
 Bacteriology Aims to Explain Infection by the Invasion 
 of the Animal Economy by Microbes Three Dis- 
 eases, Diphtheria, Cholera, Tetanus, where this Ex- 
 planation is Not Possible Discovery of Chemical 
 Vaccination Discovery of the Different Chemical 
 Poisons of Bacteria which May Produce All the 
 Symptoms of the Disease without the Intervention 
 of Microbes Actual Point of View: The Infectious 
 Disease is an Intoxication by the Poison of the 
 Pathogenic Microbe 30 
 
 SECOND PART. 
 CHAPTER V. 
 
 THE CHEMICAL NATURE OF THE BACTERIAL POISONS. 
 
 Different Conceptions of the Chemical Nature of the 
 Bacterial Poisons. First Stage: The Ptomaines 
 Differences between the Ptomaines of Selmi, of 
 Gautier, and of Brieger The Peptotoxine of Brieger 
 Researches of Salkowsky, of Bouveret and Devic 
 Researches of Bassi on Typhotoxine Works of 
 Baumann The Ptomaines are Perhaps Artificial 
 Products Opinion of S. Martin. Second Stage: 
 The Diastases Researches of Roux and Yersin 
 Criticism of the Notion of Diastases. Third 
 Stage: The Toxalbumins Work of Brieger and 
 Frankel 39 
 
VII 
 
 PAGE 
 CHAPTER VI. 
 
 THE CHEMICAL NATURE OF THE BACTERIAL POISONS 
 (CONTINUED). 
 
 Criticism of the Work of Brieger and Frankel Re- 
 searches of Proskauer and Wassermann The Mi- 
 crobe Poisons have Not yet been Prepared in a State 
 of Purity Similar Non-Microbian Poisons The 
 Work of Stillmarck Demonstration of the Albu- 
 minoid Nature of the Diphtheritic Poison Toxi- 
 cological Differentiation of the Bacterial Poisons 
 Natural and Artificial Poisons: Their Characters 
 The Chemical Vaccines The Antitoxines or Cura- 
 tive Substances Hypothesis as to the Chemical 
 Nature of the Poisons The Nucleo-albumins and the 
 Nucleines The Consequences of this Hypothesis.. . 52 
 
 CHAPTER VII. 
 THE ORIGIN OF THE MICROBIAN POISONS. 
 
 Preconceived Idea that the Poisons Can Only Come 
 from the Decomposition of Albuminoid Bodies Ex- 
 periments Already Old which Overthrow this Idea 
 Researches of Polotebnoff, Popoff, Bergmann, Schul- 
 ler Numerous Researches Made at the Laboratory 
 of Pachoutine Recent Labor of Guinochet The 
 Microbian Poisons are Not Products of Decompo- 
 sition, but the Result of Synthesis Are They Se- 
 cretions? Arguments in Favor of this Idea Argu- 
 ments which Combat it Thje Poisons are Intimately 
 Linked to the Bodies of the Bacteria The Experi- 
 ments of Cantani and of the Author The Re- 
 searches of Buchner on the Proteins Criticism of 
 these Researches All the Microbe Poisons Come 
 
VIII 
 
 PAGE 
 
 from the Bodies of Bacteria Utility of this Source 
 for the Microbes 65. 
 
 CHAPTER VIII. 
 
 ACTION OF THE BACTERIAL POISONS ON THE ANIMAL 
 ORGANISM ACCUSTOMANCE AND IMMUNITY. 
 
 Insufficiency of our Knowledge of the Mode of Action of 
 the Bacterial Poisons on the Animal Organism 
 Local Action of the Modified Poisons Selective Ac- 
 tion of Tuberculin and Malleine Destruction of 
 Poisons in the Bodies of Refractory Animals 
 Explanations of the Different Contradictions 
 Resume and Conclusions on the General Toxicity of 
 Microbes 78 
 
 THIRD PART. 
 
 CHAPTER IX. 
 THE POISONS OF TETANUS. 
 
 Researches of Brieger, Kitasato and Weyl, Knud Faber, 
 Tizzoni and Cattani, Brieger and Frankel, Vaillard 
 and Vincent and Kitasato, on the Properties of the 
 Tetanus Poison The Investigations of Bruschetini 
 and of Camara Pestana on its Diffusion in the Ani- 
 mal Body Vaccination against Tetanus: Behring 
 and Kitasato The New Notions Contributed by 
 these Authors 88 
 
 CHAPTER X. 
 
 THE POISONS OF TETANUS (CONTINUED). 
 Vaccination against Tetanus is Obtained by Tizzoni, 
 
IX 
 
 PAGE 
 Cattani, and Vaillard The Researches of Kitasato; 
 
 of Behring; of Brieger, Kitasato, and Wassermann 
 Immunization The Labors of Tizzoni, Cattani, 
 and Vaillard The Researches of Ehrlich; of Brieger 
 and Ehrlich; of Brieger and Frank Application of 
 the Method of Behring and Kitasato to the Treat- 
 ment of Tetanus in Man 97 
 
 CHAPTER XI. 
 
 
 THE POISONS OF DIPHTHERIA. 
 
 The Researches of Roux and Yersin; of Loffler C. 
 Frankel Succeeds in Vaccinating Animals against 
 Diphtheria The Researches of Behring and His 
 Discovery of Immunization The Difficulties Not 
 Yet Surmounted of Diphtheritic Vaccination 106 
 
 CHAPTER XII. 
 
 THE POISONS OF CHOLERA AND OF THE AVICIDE 
 VIBRIO. 
 
 The Researches of the Author on the Poisons of Cholera 
 and of the Avicide Vibrio The Researches of Her- 
 nandez and Brtthl, of Niessen and Behring, and of 
 Zasslein 1 14 
 
 CHAPTER XIII. 
 THE POISONS OF TUBERCULOSIS. 
 
 The Researches of Koch, of Maffuci, of Prudden and 
 Hodenpyl, of Straus and the Author, of Grancher 
 and Ledoux-Lebard The Tuberculin of Koch, and 
 the Researches which it has Provoked The Toxo- 
 mucin of Weyl 120 
 
X 
 
 PAGE 
 CHAPTER XIV. 
 
 THE POISONS OF CHARBON AND OF GLANDERS. 
 
 Insufficiency of our Toxicological Knowledge respecting 
 Charbon and Glanders Different Writings on the 
 Subject 127 
 
 CHAPTER XV. 
 
 PRACTICAL RESULTS OBTAINED BY THE STUDY OF 
 MICROBIAN TOXICOLOGY. 
 
 Rapid Indications respecting the Other Bacterial Poisons 
 
 Immunization as a Result of Toxicological Studies. 133 
 
PREFACE. 
 
 The Bacterial Poisons are here for the first time made 
 the subject of a connected treatise. The genera] plan of this 
 work is as follows: 
 
 When bacteriology, extending its domain beyond certain 
 etiological data concerning infection, sought to penetrate the 
 pathogeny thereof, there arose a necessity of undertaking 
 the study of chemical poisons produced by the microbes. This 
 study first imposed itself in diseases such as cholera, diphthe- 
 ria, and tetanus, where the pathogenic agent is found lodged 
 in a limited part of the organism and yet causes a general 
 affection. In these cases, the pathogenic action of the bac- 
 teria can be explained only by the systemic poisoning pro- 
 duced by the specific products of these bacteria. But the 
 same explanation has been found to hold good in diseases of 
 another type, where, as in tuberculosis, the lesion produced 
 by the pathogenic agent is strictly circumscribed to the spot 
 where this agent vegetates; here also we have proved that the 
 lesion does not come from the microbe as a living organism, 
 but from its chemical poisons the lesion and the disease 
 may be reproduced by the dead microbe, which can, of course, 
 act only by the toxic substances which it contains. 
 
 The study of bacterial poisons concerns not only the 
 pathogeny of infectious diseases; it finds also practical appli- 
 cations for their prophylaxis and treatment. It has been 
 demonstrated that a sure and inoffensive vaccination may be 
 obtained without the help of living microbes, and that the 
 refractory state may be conferred simply and solely by the 
 soluble products of micro-organisms. Lastly, a new method 
 has been found immunization which has direct applica- 
 
XII 
 
 tions in therapeutics, being capable not only of preventing 
 disease, but of curing it when once declared. Here the role 
 of living bacteria is even less than in chemical vaccination. 
 The cure of infectious diseases is obtained with the serum of 
 vaccinated animals. 
 
 The theory of immunity has also profited by the knowl- 
 edge of the poisons of bacteria; it has been shown that the 
 immunity of animals against infectious diseases is in close 
 relation with their resistance to microbial poisons; the latter 
 is subordinated in its turn to the elaboration by these ani- 
 mals of special substances antitoxines which have the 
 property of neutralizing the action of microbial toxines. 
 
 From this aggregate of facts, a new science has sprung 
 up: the science of Microbial Poisons, which is based at once 
 on bacteriology, on biological chemistry, and on general 
 physiology. 
 
 From bacteriology it borrows its data about microbes as 
 producers of poisons; but the important results obtained with 
 the analogous poisons of different origin, such as abrine, 
 ricine, robine, necessitate the extension of the domain of 
 microbial toxicology beyond microbism. 
 
 To biological chemistry belong the methods of prepara- 
 tion and of isolation of bacterial poisons; from it we must 
 demand information as to their nature and their production. 
 
 Lastly, the study of their mode of action on the animal 
 organism, of their diffusion, and of their destruction in the 
 organism, belongs to the domain of general physiology. 
 
 Thus, microbial toxicology touches at once on bacteriol- 
 ogy, on biological chemistry, and on general physiology; it 
 is the science of bacterial poisons, and has for its object the 
 -study of their chemical nature, of their mode of production, 
 .and of their action on animals. 
 
 This work is divided into three parts: 
 
 The first part History of Microbial Toxicology de- 
 
XIII 
 
 scribes the evolution which the study oi systemic poisoning 
 in the infections has undergone. 
 
 The second part General Toxicology treats of our 
 actual information respecting the chemical nature of the 
 microbial poisons, their production, and their destiny in the 
 animal body. 
 
 The third part Special Toxicology will be the exposi- 
 tion of the data acquired concerning the toxines of different 
 diseases. N. G. 
 
 Paris, May, 1892. 
 
THE BAOTEKIAL POISONS. 
 
 First Part: History of the Development of 
 
 Our Knowledge Respecting the 
 
 Bacterial Poisons. 
 
 CHAPTER I. 
 
 EXPERIMENTAL STUDY ON THE PUTRID 
 POISON. 
 
 SUMMARY. Experiments with the Toxic Effects Pro- 
 duced by Substances Undergoing Decomposition 
 Seybert, Gaspard, Stick Establish the Symptoms and 
 the Lesions of Experimental Septicczmia Panum 
 Gives the Proof that it is Produced by a Chemical 
 Poison Bergmann and Schmiedeberg Determine 
 This Poison to be an Alkaloid: Sepsine. 
 
 The first microbial poisons studied experimentally 
 were products of putrefaction. Chemists were led to 
 this study by the important etiological role which 
 putrefaction plays in ancient medicine. It was be- 
 lieved that the emanations from decomposing matters 
 might produce typhoid and malarial fevers. The 
 
 I 000 
 
putrefaction of wounds gave rise to the complications 
 so formidable and so frequent, of septicaemia and 
 pyaemia. A "spontaneous" tendency of the living 
 organism to putridity characterized certain wide- 
 spread diseases such, for instance, as scorbutus. 
 The diseases of putrid origin thus constituted an 
 important group of affections which corresponded 
 nearly to our infectious diseases of to-day. 
 
 Despite this importance which, from an early 
 antiquity, medicine accorded to putrefaction, the 
 experimental study of the latter is relatively recent. 
 
 Towards the end of the last century, gtejj|^.- jr 
 perimented with blood serum, pus, and putrid meat. 
 He introduced these matters into the general circula- 
 tion of the dog. He noticed that the toxic effects 
 varied proportionately to the quantities of liquid in- 
 jected. Thus, for instance, 20 cubic centirneters of 
 putrid serum killed a dog Jn_the_j^ace7flL~several 
 hours, with vomitings, convulsions, and progressive en- 
 feeblement; 3 grammes caused deathjrrtwo da^s, with 
 dysenteric pnenomefrar^ doses below two grammes 
 produced only a temporary diarrhoea, followed by 
 a complete restoration of the animal. Seybert also 
 studied the effects of the same putrescent sub- 
 stances when they were introduced by mouth into 
 the stomach. In these cases the toxic effects were 
 nil. Moreover, in opening the stomach of the dog 
 three hours after a meal of putrid meat, Seybert no 
 longer found in the mass undergoing digestion the 
 
? ..-. 
 
 putrid aspect and odor. Seybert cites in this connec- 
 tion the researches of Spalanzani, who was absolutely 
 convinced of the innocuousness of putrid foods after 
 having fed dogs, cats, and birds with them. 
 
 Gaspard, physician of Saint Etienne, pursued 
 some experimental researches on the putrid poison. 
 His experiments established several important points 
 respecting the effects of putrid matters. 
 
 y Gaspard showed that (fafiflmp 1 ^ blood, pus, 
 and meat produce in animals a rapid and mortal 
 intoxication. On the contrary, fresh organic liquids 
 such as saliva, urine, and sperm, when injected for 
 control, are found to be non-toxic. Gaspard showed, 
 moreover, that the putrid infusions of plants also 
 poison animals. 
 
 The symptoms of this poisoning were very con- 
 stant and typical, being independent of the source of 
 the putrid substances, whether vegetable or animal, 
 and of the place of the injection. This intoxication 
 (septic or putrid infection) was characterized by 
 tremblings and convulsions, vomitings, and diarrhoea, 
 the latter often sanguinolent. The animals died in 
 dyspnoea, cyanosis, and complete prostration. 
 
 At the autopsy, the pathognomonic lesion of the 
 putrid infection consisted in a haemorrhagic enteritis. 
 
 Gaspard concluded from his experiments that a 
 special poison, the putrid poison, is formed by putre- 
 faction. This poison produces the same disease and 
 the same lesions in all the animal species. 
 
4 
 
 In order to determine the nature of the putrid 
 poison, Gaspard experimented with the different 
 gases formed by putrefaction. He found that car- 
 bonic acid and sulphuretted hydrogen are not 
 
 that ammonia, on the contrary, is toxic, but that it 
 cannot produce the symptoms of putrid infection^ 
 Hence was overthrown the preconceived idea of the 
 ancients, who believed the principal danger to be in 
 the putrid gases of decomposition. The researches 
 of Gaspard established subsequently the formation, 
 in the course of putrefaction, of a special and very 
 active poison.* 
 
 Other experimenters followed in the way marked 
 out by Gaspard. Magendie, Leuret, Dupuis, Darcet, 
 and Sedillot confirmed on all the principal points the 
 fundamental reseaches of Gaspard. They discovered 
 his putrid poison, without, however, determining its- 
 chemical nature. 
 
 Virchow found that the putrid poison does not 
 act, like the other chemical poisons, in proportion to- 
 rts quantity. Its activity depends rather on the de- 
 gree of decomposition of the substances whence it is 
 derived. From this point of view, Virchow ranks it 
 among the ferments. 
 
 Stich has made very careful and interesting re- 
 'Searches on putrid matters. He injected putrefied 
 albuminoid products, after repeated filtration through- 
 
 * Gaspard: Physiological Memoir on the Purulent and. 
 Putrid Maladies (Journal de Physiologic, 1822 and 1824). 
 
T r* 
 
 paper, into the veins of dogs, of hares, and of birds. 
 The latter showed themselves the most susceptible to 
 poisoning. The typical lesion consisted in a hyper- 
 semia of the intestines. 
 
 But especially interesting are the researches of 
 Stich on the toxicity of faecal matters. The aqueous 
 extract of the excrement of a dog, injected into the 
 veins of this animal, was found to be very toxic; but 
 the same extract introduced into the stomach or rec- 
 tum of dogs was completely inoffensive. 
 
 Stich carried this study farther. Introducing 
 the excrement of one animal into the stomach or 
 rectum of an animal of another species, he found that 
 poisoning was the result. It follows that immunity 
 exists for animals only in relation to their own in- 
 testinal contents. Stich asks: What are the causes of 
 this absence of auto-intoxication?* He leaves the 
 question unsolved. 
 
 The most important researches on the putrid 
 poison belong indisputably to Panum. 
 
 J^i^rr^asks if the accidents of putrid infection 
 are really due to a chemical substance or to the action 
 of bacteria multiplying in matters undergoing putre- 
 faction? To solve this question, he instituted numer- 
 ous important researches on the putrid poison, and 
 confirmed the results obtained by Gaspard. 
 duced a typical intofijfiflt^" *T **"- '"-f^*^ nf 
 fied meat. This intoxication was characterized by 
 
 * Stich, Charite Annalen, 1853. 
 
K 
 
 prostration, vomiting and diarrhoea, and rapid death. 
 The autopsy showed a more or less pronounced 
 gastro-intestinal catarrh. 
 
 Having reproduced this putrid infection of Gas- 
 pard, Panum then demonstrated that it was due to 
 the action of a chemical poison independent of the 
 
 ^pjpO^g^^gj^ggjBjpgj^^^gjgfcMefMVHMUMMMPMMMI . ..** * 
 
 agency of bacteria. The putrid liquids, though per- 
 fectly clear, and freed from every foreign germ by re- 
 peated filtration through paper, none the less retained 
 their toxic properties. Nor was the putrid poison de- 
 stroyed by ebullition for eleven hours, which ought 
 surely to kill all living germs. Having thus settled 
 the fundamental question as to the inorganic charac- 
 ter of the putrid poison, Panum studied its chemical 
 properties. He found that this poison was not vola- 
 tile; for the distilled products of the putrefied matters,, 
 while having the fetid odor, are not at all toxic. 
 The putrid poison was fixed, and the dry residue ^of 
 the evaporation of the putrefied matters retained all 
 their toxic properties.. This dried residue did not 
 give up to alcohol its chief toxic principle. The alco- 
 holic extract had a different physiological action from 
 the putrid matters; it produced a manifest narcosis,, 
 which Panum compared to that produced by the 
 alkaloids of opium. The veritable putrid poison 
 capable of causing the typical poisoning was, on the 
 contrary, soluble in water, although it adhered to the 
 precipitates of the albuminoid substances coagulable 
 by heat; it was also partially retained by the filters. 
 
L t 7 __ __ 
 
 Panum showed also that his poison had nothing in 
 common with the well known substances produced by 
 putrefaction, such as leucin and tyrosin. He stopped 
 here in his chemical investigation, and did not further 
 indicate the nature of his putrid poison. It was suffi- 
 cient for him to have given the irrefutable proof that 
 it was not organized. 
 
 The work of Panum produced an enormous im- 
 pression upon the scientific world. Hidden by its 
 author in an unknown Danish publication, the memoir 
 of Panum was found there by a German writer, 
 who published an analysis of it in Schmitt's Jahr- 
 biicher for 1859. In this extensively circulated pub- 
 lication the results of Panum's researches became 
 known to the whole scientific world. A great num- 
 ber of experimenters in Germany and, especially, in 
 Russia, occupied themselves with the verification of 
 these results and the chemical study of the putrid 
 poison. 
 
 The University of Munich then offered a prize 
 for the best essay on " Putrid Infection and Its 
 Causes." Two memoirs were deemed worthy of a 
 prize: those of Henner and Schweninger. These 
 writers confirmed the fundamental result of Panum, 
 that the putrid poison is of chemical nature, and that 
 the action of microbes is nil in putrid infection. As 
 to the nature of the poison, they favor Virchow's 
 view that it should be ranked among the ferments, for 
 it acts in infinitesimal quantity, has a period of incu- 
 
bation, and always produces the same typical affec- 
 tion. 
 
 Since the same year, 1866, there have appeared 
 at Dorpat numerous theses devoted to the chemi- 
 cal study of the putrid poison. We may cite the 
 memoirs of Raison, Frese, Weidenbaum, Schmitz, 
 Petersen, A. Schmidt, and Brehm. These theses also 
 confirm Panum's position. Raison shows that the 
 putrid poison is not retained by filtration through 
 charcoal. The minimum dose of 0.0036 gramme of 
 the filtered liquid killed a horse by intravenous in- 
 jection. He found also that the putrid poison, evap- 
 orated to dryness, could be subjected for several 
 hours to a heat of 130 C. Weidenbaum has seen 
 that the putrid poison supports discontinuous ebulli- 
 tion, repeated several times. These two writers, and 
 the other pupils of the school of Dorpat whom we 
 have mentioned, have also seen the putrid poison 
 support without alteration the different chemical 
 manipulations, such as the action of sulphuric and 
 hydrochloric acids, precipitation by acetate of lead 
 and nitrate of mercury, etc. Lastly, they even suc- 
 ceeded in obtaining the putrid poison in a state of 
 purity, and of determining its chemical nature. It is 
 the celebrated sepsine found in 1868 by Bergmann 
 and Schmeideberg. This important result was ac- 
 complished by a complicated method based on the 
 employment of corrosive sublimate. 
 
 In 1866, Bergmann published the results of his 
 
9 
 
 researches made with Schmiedeberg on the chemical 
 properties of the poison contained in putrefied yeast. 
 They found that this poison supports for eight hours 
 digestion in boiling alcohol. The alcoholic extract 
 of the putrefied yeast is found, after evaporation, to be 
 powerfully toxic. To separate the active principle, 
 Bergmann and Schmiedeberg precipitated it in alco- 
 holic solution by corrosive sublimate. The mercurial 
 precipitate was washed in alcohol, dissolved in water, 
 and decomposed by hydrogen sulphide. The liquid, 
 freed from sulphide of mercury by filtration, was 
 heated to get rid of the excess of hydrogen sulphide, 
 and treated with carbonate of silver, which, by form- 
 ing an insoluble chloride of silver, removed free 
 hydrochloric acid from the solution. The filtered 
 liquid obtained after this last operation was almost 
 colorless and perfectly clear. It had the specific 
 toxicity of the putrid poison. 
 
 In 1868 these experimenters (Bergmann and 
 Schmiedeberg) obtained, in crystalline form, the salt 
 of this poison with sulphuric acid, and called 'it the 
 sulphate of sepsine. This salt was proved to be 
 eminently toxic. Injected in the dose of one centi- 
 gramme in the vein of the dog, it provoked imme- 
 diately vomiting and sanguinolent diarrhoea, and at 
 the autopsy hsemorrhagic ecchymoses were found in 
 the stomach and intestines. 
 
 The publication of Bergmann and Schmiedeberg's 
 discoveries naturally produced a profound impres- 
 
IO 
 
 sion. For a time, chemists believed they had at last 
 got hold of the famous putrid poison whose existence 
 had been demonstrated by Gaspard, whose purely 
 chemical nature had been proved by Panum, and which 
 had finally been obtained in a crystalline form by Berg- 
 mann and Schmiedeberg. This sepsine would alone 
 henceforth explain all the various accidents of medi- 
 cal and surgical septicaemia. It was believed that the 
 way was finally opened for the easy interpretation of 
 all the infections. Chemists went to work to seek 
 out and to study this sepsine. Petersen and A. 
 Schmidt found it in putrid blood, and thus confirmed 
 the data of Bergmann and Schmiedeberg. 
 
 Nevertheless, the most competent authorities 
 soon admitted the impossibility of ascribing all the 
 effects of the putrid poison to sepsine alone. First, it 
 is not found in all putrefied substances; thus, for in- 
 stance, Fischer could not find it in putrid pus. Berg- 
 mann himself, in applying his method of extraction 
 to different putrid matters, has isolated substances 
 which had nothing in common with sepsine. Other 
 toxic substances were extracted from the putrid 
 matters. 
 
 Zuelzer and Sonnenschein have found another 
 septic alkaloid in the infusions of putrid meat. This 
 body has all the reactions of the vegetal alkaloids, 
 such as atropine and hyoscyamine. It possesses also 
 the physiological action of the latter. Injected under 
 the skin of animals, it provokes dilatation of the 
 
pupils, relaxation of the intestines, and exaltation of 
 the heart's action. 
 
 The conclusion was inevitable that the poisons 
 produced by putrefaction were numerous and differ- 
 ent, according to conditions necessary to determine. 
 In the following chapter we shall study the researches 
 which have been undertaken to elucidate the rationale 
 of putrefaction and of septicaemia. 
 
 Only, we shall see in studying the different pos- 
 sible causes which influence the production of the 
 putrid poison, that experimenters had overlooked the 
 principal one, viz., the intervention of microbes. Be- 
 tween the Chemical School and the Vitalists, there 
 was destined to be a memorable struggle, which 
 ended in the victory of the latter a victory which 
 completely cast into the shade the results acquired by 
 the chemical study of the putrid poison. 
 
CHAPTER II. 
 
 THE MICROBIAL ETIOLOGY OF PUTREFACTION 
 AND OF THE INFECTIONS. 
 
 SUMMARY. Contest between the Chemical and the 
 Vitalist Doctrines The Contradictions and the 
 Results Obtained by the Chemists Their Explana- 
 tion Insufficient by Reason of the Absence of the 
 Notion of Specificity Definitive Victory of the 
 Vitalist Doctrine The Infectious Diseases are Ad- 
 mitted to be Caused by Specific Fermentations. 
 
 From antiquity, two contrary interpretations of 
 the etiology of putrid and infectious diseases have 
 prevailed. 
 
 The Chemical Doctrine set forth in the previous 
 chapter is, that matters in the process of decomposi- 
 tion constitute poisons sufficient to account for all the 
 symptoms of the infections, and that the microbes 
 found in these putrid matters are only inconstant and 
 inoffensive accompaniments. The other, or Germ 
 Theory, affirms that the microbes are really the sole 
 agents of putrefaction and of infectious diseases, and 
 that just as the inanimate matters do not alter and fer- 
 ment without the intervention of microbes, so the 
 presence of the latter is indispensable for the produc- 
 
13 
 
 tion of the infectious disease. We shall now see how 
 this Vitalist doctrine has definitively supplanted the 
 Chemical. 
 
 The chemical and toxicological researches on the 
 putrid poisons did not realize the fundamental condi- 
 tion of all scientific work; they did not give results 
 that were always constant and uniform. Chemical 
 analysis revealed different properties in the active 
 principles isolated. Experimentation in the hands of 
 different chemists did not produce the same symptoms 
 and the same lesions with the poisons obtained from 
 putrid substances. This inconstancy of results was 
 soon noticed by investigators. Thus, for instance, 
 one of them * expresses himself in this way: 
 
 " There is not agreement as to the pathology of 
 the putrid infection or as to its nature. Thus, cer- 
 tain chemists (Henner, Schmidt, Petersen) affirm that 
 the convulsions and tetanic attacks are important 
 characters of putrid infection; others, on the contrary 
 (Dupuis, Sedillot, Stick, Raison, and Schmitz), have 
 never or but rarely seen them. Even the constant 
 existence of vomiting and diarrhoea, supposed to be 
 pathognomonic of putrid infection, is denied by cer- 
 tain experimenters. The same may be said of the 
 anatomical lesions. Thus Gaspard, Leuret, Virchow, 
 Panum, Stick, Henner, Bergmann, and others affirm 
 
 *Rawitsch: Zur Lehre von der putriden Infection und 
 deren Beziehungen zura sogenannten Milzbrand. Berlin,. 
 1872. 
 
14 
 
 that hsemorrhagic inflammation of the intestinal 
 mucosa is a characteristic lesion; while others (Du- 
 fiuis, Sedillot, Billroth, J?aison,and Schmidt) have found 
 in mortal cases the intestinal mucous membrane 
 intact. Several writers think that sanguineous ecchy- 
 moses in the different organs are characteristic of 
 putrid infection, while others have not seen them in 
 their autopsies. Bergmann believes that the modifi- 
 cations of the spleen are constant and pathognomonic, 
 while Davaine affirms that it is just the absence of 
 hypertrophy of the spleen in putrid intoxication that 
 enables us to differentiate it from charbon. 
 
 To explain the causes of these variations in the 
 results of experimentation, writers have referred us 
 to the different sources of the putrid substances, 
 alleging that putrefied meat acts differently from 
 putrefied blood, etc. Stress has also been laid on the 
 differences of the action of the putrid poison, accord- 
 ing to the different places of its introduction into the 
 animal economy. Thus, the effects of subcutaneous in- 
 jections were distinguished from those of intravenous 
 injection; in the first case, a more or less intense 
 local inflammation was aroused, complicated with 
 suppuration and gangrene; in the second case, there 
 appeared the septic infection, as Gaspard described 
 it. We have, moreover, been referred to the different 
 receptivity of the animals under experimentation, 
 varying according to species, race, and age. But it 
 has been especially remarked that the toxicity of 
 
putrid products undergoes very considerable varia- 
 tions in the course of putrefaction. The putrefied 
 matters, instead of becoming more and more toxic as 
 decomposition went on, were found to be even less 
 toxic, the first stages only giving rise to the most 
 violent poisons. Billroth shows that relatively fresh 
 pus, even when it is not putrid but " good and laud- 
 able," may produce by intravenous injection very 
 violent and febrile reactions, even ending in death. 
 H. Fischer demonstrated that pus which was very 
 toxic at the onset, loses, in putrefying, its specific and 
 septic toxicity. Bergmann has found that putrefied 
 blood is more toxic in the first five to six days. Heller 
 has remarked the same fact. 
 
 Samuel has studied this question in detail. He 
 finds that there are three periods in the toxicity of 
 putrid substances: phlogogenous, septogenous, and 
 pyogenous. In the first, the inoculation of putrid 
 substances is followed by transient inflammatory 
 symptoms which disappear without leaving any local 
 or general troubles. In the septogenous stage, specific 
 toxic products appear which provoke the true "septic 
 infection." It is here that we note the most terrible 
 local and general phenomena of septicaemia: the 
 former going on to septic gangrene, the latter to ful- 
 minant septicaemia which kills with the rapidity of 
 hydrocyanic acid. Later on, these septic properties 
 of the progressively decomposing putrid substances 
 become attenuated little by little, to leave remaining 
 
i6 
 
 in the pyogenic stage only the power of prodncing 
 localized and benign suppurations. 
 
 But all these distinctions, despite their consider- 
 able interest, did not quite suffice to explain the in- 
 constant and conflicting results of experimenters. 
 
 There remained always an irreconcilable differ- 
 ence between the chemical poison of Panum, the activ- 
 ity of which depended on the dose injected, and the 
 virus of Davaine, which acted in infinitesimal doses and 
 which was found in increased quantity in the cadavers. 
 It was evident that the septicaemias of which the dif- 
 ferent writers spoke were not at all identical, and that 
 their differences depended on a factor having quite 
 other powers than those 'alleged by the chemists. 
 But above all, the chemical doctrine was unable to 
 explain the remarkable fact discovered by Coze and 
 Feltz, and confirmed by Davaine, Hiller, and others: 
 the septic poison, instead of being enfeebled by 
 dilution, like all the chemical poisons, in the cadavers 
 of the animals it had killed, was found to be exalted. 
 It was evidently a special poison fitted to reproduce 
 itself just like living beings. From this fact there 
 was only one step to the conclusion that this poison 
 was constituted by bacteria. One might, it is true, 
 still find refuge in the hypothesis of Liebig and 
 Robin, that the soluble ferments are albuminoid mat- 
 ters in process of decomposition, which may transmit 
 their alterability to other bodies and reproduce them- 
 selves in this way indefinitely. But this hypothesis. 
 
of Liebig was completely ruined in its chemical ap- 
 plications by Pasteur, who demonstrated peremptorily 
 that fermentation (or decomposition) does not exist 
 apart from the life of organized ferments. Moreover, 
 the contagionists were soon able to go farther in 
 showing the fallacy of the chemical doctrine, and to 
 prove that certain microbes separated from their 
 morbid products were able to produce determinate 
 diseases. Thus, it was proved that charbon, hereto- 
 fore ranked among the putrid fevers, was produced 
 by the bacterium of Davaine; that septicaemia was 
 caused in man by the septic vibrio of Pasteur; that 
 the different septicaemias and pyaemias in animals had 
 each its special pathogenic agent, living staphylococci 
 or streptococci. At the same time was established 
 the fundamental notion, proclaimed by Ferdinand 
 Cohn, of the specificity of the bacteria, which con- 
 stitute each a being apart, characterized by its mor- 
 phology and by its functions, chromogenic, zymotic, 
 or pathogenic. Weigert and Ehrlich brought to the 
 aid of this study the processes of staining by anilin 
 dyes, which enable microscopists to recognize the bac- 
 teria. Koch introduced the Abbe" method of illumina- 
 tion and the method of culture on solid media, which 
 have helped bacteriologists to make immense strides 
 in their knowledge of the microbes. Thus was con- 
 stituted Bacteriology, which has been making such 
 rapid advances. It is now known with certainty that 
 the infectious diseases are due to the action of mi- 
 crobes. 
 
i8 
 
 The vitalist doctrine was not in absolute contra- 
 diction with the results obtained by chemists, and 
 Panum has the merit of having understood, just as 
 Selmi and Brieger did later, the possibility of recon- 
 ciling the two theories. 
 
 Panum said "that it is incontestible that my 
 putrid poison is purely chemical, but it might be pro- 
 duced by a microbe, even a determined microbe. In 
 diseases, both the pathogenic microbe and the produc- 
 tion of the poison by the microbe may play their 
 respective parts." 
 
 But the conciliatory voice of Panum was not 
 heeded, and the chemists made vigorous opposition to 
 the victorious march of bacteriology. They continued 
 to bring forward proofs in favor of this view that the 
 microbes were but simple accompaniments of the 
 poisons of Panum, Bergmann, and Schmiedeberg. 
 They filtered the toxic products through filtering 
 paper or through clay, and studied the effects of these 
 filtered liquids. They employed heat, dialysis, the 
 different chemical reagents, to eliminate the living 
 microbes of their experiments. They cultivated the 
 microbes in media deprived of albuminoid substances, 
 to show that in these cases they were inoffensive 
 (Hiller). But to bacteriologists all these experiments 
 of the chemists were null and void, for they could not 
 have been well conducted without previous knowledge 
 of the properties of the bacteria. Nay, more; all the 
 experiments on systemic intoxication where the in- 
 
i 9 
 
 fluence of a chemical poison made itself felt, could 
 but embarrass the bacteriologists, adding a supple- 
 mentary factor to their study of the functions of bac- 
 teria. 
 
 Under these conditions the bacteriologists, quite 
 naturally, made a profound distinction between in- 
 toxication and infection. Intoxication was character- 
 ized by the sudden appearance of the accidents, by 
 the proportionality of the effects to the doses (doses 
 always considerable), by the absence of specificity for 
 it could be produced by the most common bacteria if 
 injected in sufficient quantity. It had to be carefully 
 avoided in bacteriological studies. These concerned 
 themselves only with infection, which was produced by 
 life and the multiplication of specific microbes in the 
 animal body. It was independent of the dose of 
 microbes introduced into the animal body, and it ap- 
 peared after an incubation necessary for their multi- 
 plication. The infectious disease was a specific 
 fermentation, linked to the life and multiplication of 
 the pathogenic microbes, just as alcoholic fermenta- 
 tion depends on the life of the yeast. 
 
 In ignoring intoxication the bacteriologists had 
 to ignore all the experimental researches on the 
 chemical poisons; and the dualism of intoxication and 
 infection long bore as a heavy weight on the develop- 
 ment of microbiology. But during this time the 
 microbian poisons reappeared in a quite different 
 science, and their study was being pursued outside 
 of bacteriology. 
 
CHAPTER III. 
 
 THE DISCOVERY OF PTOMAINES. 
 
 SUMMARY. Selmi Draws Attention to the Ptomaine? 
 Obtained from Cadavers Leucoma'ines and Pto- 
 maines of Gautier The Researches of Brieger on 
 Ptomaines. 
 
 In a celebrated trial, where the domestic of Gen. 
 Gibbone was accused of the death of the latter, the 
 chemists found in the cadaver the alkaloid delphinine. 
 But Francois Selmi, professor of chemistry at Bo- 
 logne, who was called as an expert witness for the 
 defense, demonstrated by a detailed chemical analysis 
 that the base isolated by the government experts was 
 quite distinct from the delphinine and other alkaloids 
 of vegetal origin. According to Selmi, the base in 
 question was of animal origin, and came from the 
 putrefaction of the cadaver. This dictum of Selmi 
 was completely at variance with the general convic- 
 tion, according to which the alkaloids could only be 
 produced by plants. Shortly after, similar facts were 
 adduced, and drew attention more and more to the 
 researches of Selmi. In the trial of Sonzagno, at Cre- 
 mona, the first experts thought they had found 
 morphine in the cadaver. Selmi peremptorily demon- 
 
21 
 
 strated that what they had found was a cadaveric 
 alkaloid. 
 
 There was still later a celebrated trial in Italy 
 where the prosecution endeavored to make out a case 
 of poisoning by strychnine; here Selmi affirmed the 
 cadaveric source of the base found, and the prisoner 
 was acquitted on this testimony. 
 
 Since 1870, Selmi has been making experimental 
 and chemical investigations on the organic bases 
 found in cadavers. To this order of researches he 
 was led by noting the presence in cadavers of sub- 
 stances which possessed the general and even the 
 specific reactions of alkaloids, but which could be 
 distinguished by the absence of all toxic action. 
 Selmi had the notion that alkaloids similar to those of 
 animal origin could be produced by the very fact of 
 putrefaction. To verify this idea he instituted a long 
 series of researches upon exhumed cadavers, of which 
 the disease, the death, and the duration of burial were 
 known to him. He there discovered, in fact, a great 
 number of both inoffensive and toxic alkaloids, with 
 different chemical properties, and more or less like 
 the alkaloids of plants. 
 
 Pursuing further his researches on these products, 
 Selmi demonstrated their origin in the putrefaction of 
 albuminoid matters, for he found that ptomaines also 
 take rise in the albumin of putrefied egg. 
 
 These discoveries of Selmi were too revolutionary 
 of former notions to be accepted without opposition. 
 
22 
 
 We have already mentioned how, in certain criminal 
 prosecutions, they forced themselves upon the atten- 
 tion of the whole scientific world. 
 
 These expert findings of Selmi in Italy had also 
 their counterpart in Germany, in the celebrated trial 
 of Brandes-Klebs in 1874. Chemists had extracted 
 from the cadaver of Klebs a very toxic alkaloid which 
 they had identified with coniine. Otto showed that 
 it was neither coniine nor nicotine, though it resembled 
 these alkaloids. He believed it to be a ptomaine,, 
 and this was the opinion of the experts. 
 
 It is doubtless true that Selmi was not the first to 
 find organic bases of animal origin; that Marquardt, 
 of Stettin, in 1865 extracted from a human cadaver a 
 toxic alkaloid resembling coniine, which he called 
 septicine; that Dupr and Bence Jones, in 1868, 
 found in animal bodies a substance bearing a marked 
 resemblance to an alkaloid; that Sonnenschein, in 
 1869, discovered in a cadaver an alkaloid which he 
 thought was coniine, but which proved not to be toxic; 
 the merit none the less belongs to Selmi of having been 
 the first to subject these animal bases to a profound 
 series of researches, and of thereby explaining their 
 true meaning and importance. It is noteworthy that 
 in Selmi's estimation the interest in the study of 
 ptomaines was by no means limited to legal medicine 
 by reason of their simulation of vegetal alkaloids. 
 He had well comprehended the signification which 
 they might have for biological chemistry and physio- 
 
23 
 
 logical pathology. In biological chemistry the pto- 
 maines were to Selmi a new demonstration of the 
 absence of any clear-cut limits between the vegetable 
 and animal kingdoms. At the expense of animal 
 matters might be produced alkaloids quite like those 
 which down to Selmi's time were believed to be exclu- 
 sively of vegetal origin. To general pathology the 
 importance of the discoveries of Selmi was still 
 greater. The same substances which the microbes 
 may produce at the expense of the dead animal body, 
 they may elaborate in the living body; in other 
 words, the infectious disease produced by the microbes 
 may give rise to the formation of ptomaines in the dis- 
 eased organism. In fact, Selmi found ptomaines in 
 the urine in cases of typhoid fever, pneumonia, and 
 tetanus. The premature death of Selmi arrested these 
 important researches. 
 
 The movement imparted to science by Selmi was 
 of very great importance. A large number of memoirs 
 have been recently published on the ptomaines. To 
 mention only a few, Nencki in 1876 gave the first 
 analysis of a ptomaine, obtained by the putrefaction 
 of gelatin with the pancreas. This was collidine, a 
 base already extracted from the products of distilla- 
 tion of animal oil. 
 
 Morrigia and Battistini in 1875 showed that very 
 toxic ptomaines can be got from cadavers, whose action 
 is quite similar to that of curare. Rorsch and Fass- 
 bender found in the cadaver a ptomaine resembling 
 
24 
 
 digitalin in its action. Brouardel and Bontmy dis- 
 covered a base very similar to veratrine. Gautier 
 and Etard, in 1881, defined chemically two other 
 bases, parvoline and hydrocollidine, found in the 
 products of decomposition of fish, and of the meat of 
 the horse and the ox. The fourth ptomaine defined 
 was corindine, found in 1883 by Quareschi and Mosso. 
 
 We must devote more time to the extremely im- 
 portant researches of Armand Gautier. 
 
 Gautier had announced in 1871 that during the 
 putrefaction of fibrin there is formed a certain 
 quantity of alkaloids, both fixed and volatile. In 
 1881-1883 he made, with Etard, a series of researches 
 which disclosed the chemical nature of certain pto- 
 maines of putrefaction belonging to the pyridic and 
 hydropyridic series. 
 
 According to Gautier, all the decompositions of 
 albuminoid bodies taking place away from the air 
 give rise to alkaloids. Putrefaction is always accom- 
 panied by the formation of ptomaines; but the normal 
 life of animals, which in great part goes on in the ab- 
 sence of free oxygen, leads also to the formation of 
 organic bases, which Gautier has grouped under the 
 name of leucomaines. Such are xanthine, creatine, 
 etc. Whenever perturbations of the normal con- 
 ditions of life augment the want of oxygen in the 
 body, as in anaemia and the infectious diseases, then 
 the basic products or ptomaines arise in increased 
 quantity. The non-elimination of the normal leuco- 
 
25 
 
 maines, and the production ol ptomaines, are so many 
 Causes of intoxication (or poisonjnp^ of the 
 
 Certain leucomaines are extremely toxic; for instance, 
 the secretions of the different venomous animals. 
 (Already, in 1866, Zalewsky had isolated from the 
 salamander the alkaloid samandrinej) 
 
 We have seen with what amplitude of view Gau- 
 tier established his conception as to the origin, virtu- 
 ally identical, of the alkaloids in vegetables, animals, 
 and microbes. These bases of the same nature are 
 derived everywhere from the same source, the fllhij- 
 mins; everywhere in the same fundamental condi- 
 tions^ d ^2^^ M !?. JjlL^tfaCL .absencfer ^ g^no^ 
 quantity of oxygen. Guided by these general ideas, 
 Gauffer and his pupils found a great number of new 
 leucomaines. But the exposition of their history does 
 not enter into the plan of this book. 
 
 The study of the ptomaines properly so called, 
 which interest us particularly here, owes its great 
 progress during the past few years to the researches 
 of Brieger. Brieger has made a systematic study of a 
 great number of questions called up by the history of 
 the ptomaines. Thanks to the perfected chemical 
 methods, he has succeeded in isolating numerous 
 ptomaines perfectly denned as chemical units. We 
 shall indicate the principal results which he obtained. 
 Seeing the facility with which the ptomaines form in 
 the midst of albuminoid substances in putrefaction, 
 Brieger asked if the alkaloids might not result from 
 
26 
 
 the simple solution of the albumins by the soluble 
 ferments. In fact, in digesting meat by pepsin, he 
 noted the appearance among the products of this 
 digestion of a convulsivant organic base, which he 
 called peptotoxine. We shall return later to this 
 remarkable result, which has an enormous importance 
 in connection with this question of ptomaines. 
 
 In putrid cadavers, and in the flesh of mammals 
 and of putrid fish, Brieger found a certain number of 
 bases regarded by him as constant representatives of 
 the ptomaines of putrefaction. They all belong to 
 the category of ammoniacal compounds, amines. Be- 
 sides the methyl and ethyl amines, Brieger has isolated 
 in a state of perfect purity cadaverine, putrescine, 
 neurine, muscarine, ethylendiamine, saprine, myda- 
 leine, and a great number of others. 
 
 They are chiefly diamines. Lastly, Brieger took 
 up again the idea of Panum and of Selmi, and made 
 a study of the ptomaines produced by the microbes 
 of infectious diseases. He has studied also the bases 
 formed in the cultures by the typhic and tetanic 
 bacilli, by the vibrio of cholera. But by these latter 
 studies Brieger enters into the new period of the 
 evolution of ideas concerning the rdle of bacterial 
 poisons, a period which shall be set forth in the fol- 
 lowing chapter. 
 
 A. On the Constitution of the Artificial and 
 Natural Alkaloids. 
 
 The first vegetable alkaloid, morphine, was dis- 
 
_ 27 
 
 covered by Sertiirner in 1806. Since then the num- 
 ber of organic bases extracted from plants or pre- 
 pared artificially has been considerable, especially 
 during the past few years, and chemists have at last 
 succeeded in determining their constitution. 
 
 The principal alkaloids are derivatives of pyri- 
 dine, which much resembles benzol. 
 
 Pyridine is a benzol where an atom of nitrogen 
 (N, which is trivalent) replaces the group CH (which 
 has also three free atomicities). 
 
 In the formula of pyridine, the atoms of hydrogen 
 may be replaced by the monovalent groups 
 
 C 
 
 H- | -H, or (CH.) 
 H 
 
 Thus we have picoline. If this substitution be made 
 twice, we shall have putidine; with three groups of 
 CH 3 instead of 3 atoms of H, we obtain collidine, 
 and after a new substitution, corindine. 
 
 All these products are energetic bases obtained 
 artificially by the dry distillation of bones (oil of 
 Dippel). Collidine and corindine are already known 
 to us, for they have also been found in the products 
 of putrefaction. 
 
 The natural alkaloids of plants are a little more 
 complex, but they are also the substitution products 
 of pyridine or of piperidine (which is hydrogenated 
 pyrodine). 
 
 Thus, coniine is a propylpiperidine, or a piperidine 
 
28 
 
 where an atom of H is replaced by the monoatomic 
 group C 3 H 7 which is called propyl. Other alkaloids 
 of plants are derivatives of quinoline, which corre- 
 sponds to naphthaline where an atom ojE. N replaces 
 the group CH.* 
 
 B. The Amines. 
 
 Ammonia has the formula: 
 
 H 
 
 NH 8 , or N H. 
 H 
 
 The atoms of H in ammonia may be replaced by the 
 alcohol radicals. This substitution gives rise to the 
 ammoniacal compounds, or the amines. 
 
 Thus, methyl-alcohol has the formula CH 3 HO. 
 In this formula CH 3 is the alcohol radical called 
 methyl, combined with the group HO, which is called 
 hydroxyle. The methyl may replace one, two, or 
 three atoms of hydrogen in the ammonia. We shall 
 have monomethylamine, dimethylamine, trimethyl- 
 
 amine: 
 
 H H CH 8 
 
 N H N CH 8 N CH 3 
 
 CH 8 CH 8 CH 8 
 
 The bivalent alcohol-radicals may unite with two 
 molecules of ammonia. The resulting bodies are 
 called diamines. Thus, ethylen C 2 H 4 which has 
 the two atomicities free, forms ethylendiamine: 
 
 -C 8 H 4 
 N HH N 
 HH 
 
 * Von Pictet: The Chemical Constitution of the Vegetal 
 Alkaloids. Paris, 1888. 
 
2 9 
 
 Most of Brieger's ptomaines are diamines. Thus, 
 putrescine is a tetramethylendiamine; the bivalent 
 alcohol-radical CH 2 is called methylen: 
 
 H H 
 
 N CH 8 CH 8 CH 2 CH 8 N 
 -H H 
 
 Cadaverine is a pentamethylendiamine: 
 
 H H 
 
 N CH 2 CH 8 CH 2 CH 2 CH 2 CH 8 N 
 H H 
 
 REFERENCES. 
 
 Selmi: "Ptomaine od. Alcaloide Cadaverice." Bo- 
 logna, 1881. 
 
 Husemann: "Die Ptomaine und ihre Bedentung fttr 
 die Gerichtliche Chemie und Toxicologie " (Archiv der 
 Pharmacie, Bd. 16, 17, 19, 20, and 21). 
 
 (Effinger: " Die Ptomaine oder Cadaver Alcaloide." 
 Wiesbaden, 1885. 
 
 Gautier: " On the Alkaloids Derived from the Destruc- 
 tion of Animal Tissues, Ptomaines and Leucomalnes " (Bull, 
 de 1'Acad. de M6d., Jan. 12 and 16, 1886). 
 
 Gautier: " Chimie Biologique." Paris, 1892. 
 
 Hogouneq: " The Alkaloids of Animal Origin." Paris, 
 1886. 
 
 Roussy: " Ptomaines and Leucomalnes " (Rev. des Sc. 
 Med., t. xxxi, 1888). 
 
 Brieger: " Untersuchungen uber Ptomaine." Three 
 volumes. Berlin, 1885-1886. 
 
 Brieger: " Beitrag zur Kentniss der Zusammensetzung 
 des Mytilotoxins nebst einer Uebersicht der Bisher in ihrer 
 Haupteigenschaften bekannten Ptomaine und Toxine '" 
 (Virchow's Archiv f. Pathol. Anat., 1889, t. 115, p. 483). 
 
CHAPTER IV. 
 
 INFECTION IS AN INTOXICATION BY THE Ml- 
 CROBIAN POISON. 
 
 SUMMARY. Bacteriology Aims to Explain Infection by 
 the Invasion of the Animal Economy by Microbes 
 Three Diseases, Diphtheria, Cholera, Tetanus, 
 where this Explanation is Not Possible Discovery 
 of Chemical Vaccination Discovery of the Different 
 Chemical Poisons of Bacteria which May Produce 
 All the Symptoms of the Disease without the Inter- 
 vention of Microbes Actual Point of View: The 
 Infectious Disease is an Intoxication by the Poison 
 of the Pathogenic Microbe. 
 
 We quit bacteriology at the moment when, obtain- 
 ing a decisive victory over the chemical etiology of 
 the infectious diseases, it denied the importance of 
 the poisons produced by the microbes. The faculty 
 of the bacteria of fabricating poisons presented no 
 interest to the bacteriologists, for it belonged to the 
 common bacteria, those of putrefaction, which live on 
 materials deprived of life, while the pathogenic bac- 
 teria were characterized by their power to develop in 
 the living organism and to invade it. 
 
 We must remember that bacteriology originated 
 
in the study of fermentation, with which naturally 
 the infectious disease came to be assimilated. As 
 alcoholic fermentation had found its provisional ex- 
 planation in the life without air of the yeast plant, 
 so the disease was explained by the life of the microbe 
 in the animal body. 
 
 Thus, for example, in the deadly anthrax, whose 
 study has been intimately associated with the begin- 
 nings of bacteriology, and which long served as the 
 type of the bacterial diseases, the animals died with a 
 great quantity of bacteria in the blood of all the 
 organs. It was suppesed that the death of the ani- 
 mals was caused by this enormous mass of foreign 
 bodies invading the animal humors, and the mechan- 
 ism of the disease was explained by the mechanical 
 effects of the presence of these foreign bodies in the 
 blood, and the capillary emboli therefrom resulting. 
 Writers also insisted on the evils resulting from the 
 subtraction of oxygen from the red globules by the 
 anthracoid bacteria, the lack of oxygen in the tissues, 
 and the asphyxia therefrom resulting. They also had 
 much to say about the asphyxia of the animal by the 
 carbonic acid set free by the microbes. 
 
 In a more general way they directed our atten- 
 tion to the energetic nutrition of the bacteria, which 
 get possession of the pabulum of the animal cells and 
 kill them when starved. 
 
 Oftener still they contented themselves with 
 vague expressions about the struggle for existence 
 
3 2 
 
 between the animal cell and the microbe, without 
 attempting further to analyze this vast conception.* 
 
 This was, we repeat, because the rising bacteri- 
 ology was dominated by the analogy between the in- 
 fectious disease and fermentation by figured bodies. 
 Just as the decomposition of matters in fermentation 
 was produced not by a chemical substance, or enzyme, 
 but by the life without air of the ferments, so the 
 infectious disease was the corollary of the life of 
 the pathogenic bacteria in the animal body. And we 
 have just seen that absorption of oxygen by the bac- 
 teria was especially invoked in explanation of the 
 mechanism of the disease. 
 
 At the same time, the idea of intoxication by 
 chemical poisons, which at an early period dominated 
 infection, was not completely given up. Davaine be- 
 lieved that the anthracoid bacteria secrete a product 
 which agglutinates the red globules of the blood. 
 Pasteur, by the filtration of anthracoid cultures, proved 
 the existence of this agglutinating diastase. Toussaint 
 invoked the existence of a phlogistic poison produced 
 by the anthracoid bacteria. He believed that he 
 could even isolate it and confer with it immunity 
 against charbon. Chauveau also adduced arguments 
 in favor of this chemical vaccination by the bacterial 
 poisons. 
 
 But chemical vaccination found a formidable op- 
 
 *Duclaux: " Le Microbe et la Maladie." Paris, 1866. 
 
33 
 
 ponent in Pasteur, who saw in it the overthrow of his 
 vitalist doctrine of fermentation and disease. As in 
 the fermentations the diastases or soluble ferments 
 play only a secondary role in preparing the substances 
 for the principal transformations effected by the living 
 ferments, so the diastases of the pathogenic bacteria 
 can only serve to explain the relatively unimportant 
 symptoms of the disease which itself, as well as the 
 chemical vaccine, was due solely to the life of the 
 bacteria. 
 
 But in proportion as bacteriology made its new 
 conquests, the insufficiency of this pathogeny of the 
 infectious diseases became the more manifest. 
 
 In 1884, Loffler found the bacillus of diphtheria. 
 He showed that this bacillus is always strictly localized 
 in man to the mucous membranes which are the seat of 
 the lesion. So in animals we only find it at the site 
 of inoculation, and not in the internal organs, where 
 it cannot live. 
 
 This microbe, which does not invade the organ- 
 ism, is nevertheless very pathogenic. Loffler showed 
 that this pathogenic action is due to the production 
 by the bacillus of a violent chemical poison, which is 
 absorbed at the place where the bacillus makes its 
 habitat and vegetates. 
 
 The problem of microbian poisons still more im- 
 peratively urged itself on the attention of the profes- 
 sion when Koch discovered his cholera- vibrio. 
 
 Koch demonstrated that the comma bacillus does 
 3 ooo 
 
34 
 
 not penetrate the internal organs of the victims of 
 cholera, but remains always limited to the intestinal 
 canal. It, however, produces the grave symptoms of 
 cholera poisoning. Koch concluded that the comma 
 bacillus secretes a violent poison which explains the 
 clinical phenomena of the disease. His prevision 
 turns out to be supported by experiment. Cholera 
 cultures sterilized in totality, or filtered of bacteria, 
 and even the ptomaines extracted from these cultures, 
 have been found endowed with a toxicity more or 
 less appreciable. 
 
 A third bacterial disease was soon found to be 
 absolutely inexplicable without the intervention of a 
 soluble poison. This is tetanus. The bacillus of 
 tetanus, discovered by Nicolaier, is strictly limited to 
 the place of its inoculation. It nevertheless produces 
 a terrible constitutional intoxication. Brieger sought 
 for the tetanic poison, and found in the cultures of 
 the bacillus and in the tetanized muscles several 
 specific ptomaines which provoked convulsions in 
 animals. 
 
 The great importance of the microbial poisons 
 was emphatically demonstrated, thanks to the triumph 
 of the doctrine of chemical vaccines. 
 
 The possibility of vaccinating animals against 
 infection by the soluble products of bacteria, affirmed 
 in the case of charbon by Toussaint and by Chauveau, 
 and denied by Pasteur, was established several years 
 later by the researches of numerous experimenters 
 
35 
 
 e.g., Wooldridge for charbon, Salmon and Smith for hog 
 cholera, Beumer and Peiper for typhoid fever, Char- 
 rin for the pyocyanic disease. Latterly researches on 
 chemical vaccinations have multiplied, and we may 
 without exaggeration affirm that the acquisition of 
 immunity in infectious diseases is a fact, thanks to 
 the chemical products of the microbe. Lastly, we 
 have succeeded in separating the microbes from their 
 chemical poisons by filtration. 
 
 The decisive step in this direction was taken by 
 Roux and Yersin. They found that after filtering 
 through the Chamberland filter the cultures of the 
 diphtheria bacillus, they obtained, with the filtered 
 liquid entirely deprived of bacilli, toxic effects on the 
 animals under experimentation. These toxic effects 
 are obtained with minimum doses, and resemble the 
 infection after incubation or after inoculation with 
 the living bacilli. We may, for example, produce 
 by the injection of filtered cultures the same diph- 
 theritic paralyses as with the bacillus itself. This 
 identity of effects constitutes a real demonstration of 
 Loffler's notion that the diphtheritic bacilli act on the 
 animal economy by the poison which they produce. 
 
 The researches of Roux and Yersin were soon 
 confirmed by Loffler himself and by many other 
 experimenters. 
 
 It was also found that diphtheria is not the only 
 disease producible by the chemical products of the 
 specific micro-organism. Filtered cultures of the 
 
- 36- 
 
 bacilli of tetanus were found by Knud-Faber, Tiz- 
 zoni, Vincent, and others, to be extremely toxic. 
 These filtration products, injected in animals, re- 
 produced in them all the symptoms so characteristic 
 of tetanus. Lastly, in the case of cholera also, I have 
 succeeded in reproducing all the typical symptoms of 
 the disease with the cultures of Koch's vibrio steril- 
 ized by discontinuous heat. From many other microbes, 
 as we shall see as we go on, poisons more or less viru- 
 lent have been separated. 
 
 At the onset of this period of the discovery of 
 bacterial poisons, it was believed that we should have 
 to distinguish two classes of infectious diseases. In 
 one class would be ranged the infectant microbes 
 which produce the disease and kill solely by their 
 abundant development in the body of the animal; 
 as types of this form, we were referred to charbon, 
 tuberculosis, pneumonia, septicaemia in the hare. 
 These microbes were supposed not to have the power 
 to fabricate poisons. In the second class we had the 
 toxine-making microbes, as those of cholera, tetanus,, 
 diphtheria, which cannot pullulate abundantly in the 
 animal economy, but are endowed with extreme toxi- 
 genous power. 
 
 But this distinction is not legitimate. First, the 
 same microbes produce in certain animal species gen- 
 eral septicjmaladies, and local lesions in other species: 
 the anthracoid bacterium and the lanceolate strepto- 
 coccus of pneumonia are examples. A still more im- 
 
37 
 
 portant argument against this distinction is that in 
 the case of the so-called infectant microbes, we have 
 succeeded in finding the poisons which they prepare. 
 Thus the anthracoid poison has been described by 
 Martin and Christmas; the pneumonic poison by G. 
 -and F. Klemperer; the tuberculous poisons by Koch, 
 Maffuci, Prudden and Hoddenpyl, Strauss, and my- 
 self. 
 
 By all the researches which we have reported in 
 this chapter, it has been established to a certainty that 
 the pathogenic microbes act on the animal organism 
 by means of the poisons which they produce. From 
 this point of view the importance of the microbian 
 poisons becomes extremely great. 
 
 At the outset of our historical survey we saw that 
 systemic intoxication, to Hiller and his partisans, 
 dominated infection; the poisons " formed in the midst 
 of the tissues in decomposition" were everything, 
 and the microbes nothing, in the disease. 
 
 Later the importance of these two factors was 
 inverted: the intoxication was a common place phe- 
 nomenon, meriting no special attention; the entire dis- 
 ease was produced by the progressive invasion of the 
 living animal by the microbe. 
 
 To day intoxication has reconquered its rights. 
 Infection is considered as an intoxication, but it is a 
 special intoxication by the specific poison of a patho- 
 genic bacterium. 
 
 Progress, as Hegel has defined it, traverses three 
 
_ 3 8 - 
 
 successive stages. The second is the negation of the 
 first. The last approaches the first in its form, but 
 explains the contradiction of the first two phases. 
 
Second Part: General Microbial Toxi- 
 cology. 
 
 CHAPTER V. 
 
 THE CHEMICAL NATURE OF THE BACTERIAL 
 POISONS.' 
 
 SUMMARY. Different Conceptions of the Chemical Na- 
 ture of the Bacterial Poisons. First Stage: The 
 Ptomaines Differences between the Ptomaines of 
 Selmi, of Gautier, and of Brieger The Pepto- 
 toxine of Brieger Researches of Salkowsky, of 
 Bouveret and Devic Researches of Bassi on 
 Typhotoxine Work of Baumann The Ptomaines 
 are Perhaps Artificial Products Opinion of S. 
 Martin. Second Stage: The Diastases Researches 
 of Roux and Yersin Criticism of the Notion of 
 Diastases. Third Stage: The Toxalbumins 
 Work of Brieger and Frdnkel. 
 
 The conceptions of the bacteriologists on the 
 chemical nature of the bacterial poisons have already 
 gone through several successive stages, under the in- 
 fluence principally of other toxicological researches 
 of the same epoch. 
 
40 
 
 Panum, whose brilliant achievements deserve so 
 often to be cited, had found that the principal poison 
 of putrid matters is insoluble in alcohol, and re- 
 sembles certain albuminoid substances which, like 
 the peptones, are not modified by boiling. But since 
 Panum's researches, the attention of investigators has 
 been directed more and more to toxic substances 
 which are soluble in alcohol and are analogous to the 
 most formidable of known poisons, the vegetal alka- 
 loids. 
 
 Of the putrid poison of Gaspard and of Panum, 
 Selmi, Gautier, and Brieger studied only one part, 
 the ptomaines. Regarding their task from a purely 
 chemical point of view, these three chemists bestowed 
 but little study on the physiological action of the 
 toxic substances they were testing. Nor did they 
 compare, as Panum did, the toxicity of the pri- 
 mary poison with that of the different extracts ob- 
 tained from it. The sole end of Selmi, Gautier, and 
 Brieger was to extract from the putrid matters in a 
 state of perfect purity certain definite chemical bodies, 
 the ptomaines, whose pre-existence in these matters 
 did not seem to them to merit a special demonstra- 
 tion. All three have succeeded in their efforts. 
 
 But it would be very instructive to enter a little 
 more into detail in the examination of the results 
 which they have obtained, and to compare the differ- 
 ent ptomaines of these three writers. 
 
 Selmi found some cadaveric alkaloids extremely 
 
like those of vegetal origin. Thus, for instance, he 
 has isolated a cadaveric coniine, which by all its 
 chemical reactions and by its physiological action is 
 not distinguishable from the vegetal alkaloid. 
 
 The other chemists, contemporaries of Selmi, 
 found also, as we have already seen, ptomaines re- 
 markably like the veritable alkaloids. We need only 
 mention the bases which resemble conicine (Sonnen- 
 schein), nicotine (Wolkenhaar), atropine (Tuelzer), 
 veratrine (Brouardel and Bontmy). 
 
 Later Gautier, Nencki, Quareschi, and Mosso 
 found other ptomaines which constantly accompany 
 putrefaction. These were collidine, parvoline, corin- 
 dine bodies less complex than the vegetal alkaloids, 
 but still belonging to the pyridic series. 
 
 Later still, Brieger studied putrefaction under its 
 various aspects. He discovered everywhere new 
 ptomaines, but did not succeed in finding those of 
 Selmi, nor even those of Gautier. The ptomaines of 
 Brieger are diamines, and do not belong to the pyridic 
 series. 
 
 To what can these constant differences be due, 
 and why have these chemists constantly found these 
 different products? 
 
 Evidently, we must hold in suspicion their differ- 
 ent methods of extraction of ptomaines. We might 
 even be warranted in believing that these methods 
 were likely to create artificial products which did not 
 pre-exist in the matters analyzed. We cannot here 
 
42 
 
 enter into the chemical details as to the methods em- 
 ployed by Selmi, Gautier, and Brieger; we will con- 
 tent ourselves with the analysis of only one special 
 point of this question. We will study the history of 
 the peptotoxine of Brieger. This history has a capital 
 importance in connection with the entire question of 
 ptomaines and their origin in albuminoid matters. 
 
 We have already seen (see Chap/ III) that Brieger 
 found a toxic ptomaine among the products of peptic 
 digestion. In subjecting the different species of albu- 
 min, or even of peptone, to digestion with gastric juice, 
 Brieger discovered a toxic body soluble' in ethyl and 
 amyl alcohol, giving the characteristic reaction of the 
 alkaloids. Brieger gave the name of peptotoxine to 
 this ptomaine, which he supposed to be an aromatic 
 amine. As all albuminoid matters in undergoing de- 
 composition begin by being converted into peptones, 
 it is quite natural that Brieger should find his pepto- 
 toxine at the onset of all the putrefactions which he 
 studied. It disappeared after the first week. We see 
 what great interest is attached to peptotoxine. This 
 toxic ptomaine is the necessary stage through which 
 pass all the albumins in being liquefied by the soluble 
 ferments of animal or bacterial origin. Being formed 
 at the expense of the albumins by the relatively simple 
 mechanism of diastasic fermentation, peptotoxine 
 might well serve as a key to explain the appearance 
 of ptomaines with the intervention of microbes. 
 
 Salkowsky, who has studied with much care this 
 
43 
 
 question of the peptotoxine of Brieger, has never been 
 able to find it in the products of gastric digestion. 
 He supposed that Brieger's alkaloid was produced by 
 the intervention of putrefaction during and with the 
 aid of the manipulations.* The question has just re- 
 ceived an unexpected solution by the researches of 
 Bouveret and Devic. f These writers have demon- 
 strated that peptotoxine is an artificial product which 
 is formed at the expense of albuminoid matters by the 
 combined action of hydrochloric acid and of alcohol. 
 Already Tanret had insisted on the close kinship 
 between the peptones and the alkaloids. He had 
 even shown that by the action of soda on the peptones, 
 we may form alkaloids extractable by ether. Later, 
 Dreschel saw that albuminoid substances boiled 
 with acids yield among the products of their de- 
 composition the organic bases. Bouveret and Devic 
 have now shown that even less energetic reagents 
 cause peptotoxine to appear at the expense of the 
 proteids. The evaporation of albuminoid substances 
 at 39 C. in presence of free HC1, leads to the for- 
 mation of bodies which by the action of alcohol give 
 rise to peptotoxine. There forms, by heating with a 
 feeble excess of the free acid, an intermediate body at 
 the expense of the soluble albumins. This intermedi- 
 ate body (santonine or acid albumin?) is decomposed 
 by alcohol, and yields to the latter the toxic ptomaine 
 
 * Salkowsky, Virchow's Archiv, 1891, t. 124, p 409. 
 f Revue de Med., 1892, Nos. i and 2. 
 
44 
 
 which did not exist before the action of acid and alco- 
 hol. These important researches throw a new light 
 on the question of ptomaines. 
 
 The method of Brieger, employed in all his re- 
 searches, begins ordinarily by the evaporation of the 
 first matters in presence of hydrochloric acid, and 
 by the extraction of the residue by alcohol. Now 
 this initial operation already gives rise to the artificial 
 production of ptomaines. It follows that the results 
 obtained by Brieger can give us no information as to 
 the poisons which pre-exist in the matters subjected to 
 his investigation. They disclose to us only the prod- 
 ucts, more or less constant, of decomposition of these 
 matters as a result of varied and complicated re- 
 actions. And even, sometimes, we do not know 
 whence come these products: from the normal albu- 
 mins or from microbian poisons? Thus, for instance, 
 although Kulneff found, by Brieger's method, ethyl- 
 endiamine in the contents of a dilated stomach, we 
 believe with Bouveret and Devic that this body was 
 introduced by the operations. 
 
 What must we think of numerous other ptomaines 
 found everywhere by Brieger ? 
 
 As for the typhotoxine of Brieger, Bassi has 
 shown that it does not pre-exist in the cultures of the 
 typhoid bacillus, but that it may be extracted from 
 them by the method of Brieger.* As it has not been 
 
 * Bassi: La Tifotoxine di Brieger (Gaz. Chim. Italiana, 
 1889, t. 18, p. 521). 
 
45 
 
 obtained by the culture of any other microbe, it is 
 possible that it may be a product of decomposition of 
 the typhoid poison. 
 
 In tetanus, Kitasato and Weyl have found by the 
 method of Brieger the different ptomaines which he 
 had indicated. It is possible that these ptomaines 
 may be formed by the decomposition of the primary 
 microbian poison, and it is certain that they are not 
 the poison itself.* 
 
 More recently, Baumann has found a new method 
 for the detection of diamines. By this method he 
 has rediscovered in cystinuria many of Brieger's 
 ptomaines, and has thus put beyond doubt their inde- 
 pendent existence in the urine. But Baumann's 
 method, which is based on the employment of chloride 
 of benzoyle, is too harsh for the microbian poisons, 
 which are, as we shall see farther on, extremely fra- 
 gile.f 
 
 We see, then, that all these researches on the 
 ptomaines cannot give us any idea as to the chemical 
 nature of the microbian poisons. 
 
 First, they depend on methods which are too- 
 energetic, which may form artificial products even 
 with the normal albumins bodies relatively stable. 
 
 * Kitasato and Weyl, Zeitschrift f. Hygiene, t. viii, p. 
 404. 
 
 f Udransky and Baumann: Ueber das Vorkommen von 
 Diaminen, etc. (Zeitschrift f. Physiol. Chemie, 1889, * 13, p. 
 562). 
 
- 46 - 
 
 And one has no idea of the decompositions which 
 they may effect in the microbian poisons. 
 
 Secondly, these researches have been made too 
 exclusively from the point of view of pure chemistry; 
 while the useful study of the microbe poisons is im- 
 possible without the constant control of experimenta- 
 tion. Naught but experimentation on animals can 
 enlighten us as to the toxic effects proper to the 
 poisons of the different microbes. This alone can 
 indicate whether, after the different chemical man- 
 ipulations, we have not destroyed or decomposed the 
 primitive microbian poison, or substituted another for 
 it. This true method of microbian toxicology has 
 not been followed by Brieger and other seekers after 
 ptomaines. 
 
 We may note here that Martin, in studying the 
 poison of charbon,* comes to the same conclusion as 
 ourselves; to wit, that the ptomaine which one finds 
 in the anthracoid cultures does not pre-exist there, 
 but is there combined with an albuminoid substance. 
 
 On the other side, microbian toxicology has suc- 
 ceeded in showing, in accordance with Panum, that 
 the toxic products of bacteria are not soluble in 
 alcohol. 
 
 Arloing has extracted from cultures in bouillon 
 of the pneumococcus liquefaciens bovis a toxic phlogo- 
 genous substance which is precipitable by alcohol, 
 
 * See also in this connection, Seams Woodhead, "Bac- 
 teria and their Products," London, 1891. 
 
47 
 
 soluble in water and glycerin, and whose toxicity is 
 destroyed by heating above 110 C. Arloing gave 
 the name of diastase to this toxic substance, solely 
 because among the microbian products only the 
 ptomaines and diastases were known. The ^ former 
 are soluble in alcohol, the latter are not.* 
 
 Shortly afterward, Christmas also observed that 
 the staphylococcus aureus produces in cultures a basic 
 substance precipitable by alcohol. 
 
 Roux and Yersin experimented with the filtrate 
 of diphtheria cultures (obtained through the Cham- 
 berland filter), and published several interesting re- 
 actions of the same. This poison is enfeebled or de- 
 stroyed by heating above 60 C. It is not soluble in 
 alcohol. It is carried down by the different precipi- 
 tates which form in the midst of the liquid which 
 contains it; e.g., under the action of phosphate of 
 lime, alumina, etc. 
 
 Roux and Yersin conclude that by all these re- 
 actions the diphtheritic poison resembles diastases; 
 and none the less by its intense activity in infinitesi- 
 mal doses. 
 
 This identification of the microbian poisons with 
 the diastases was neither original nor new, for it had 
 long been a habit of chemists to attribute to diastases 
 every mysterious and unknown action exercised by a 
 substance of indeterminate nature. Thus we have 
 seen that Virchow, Schwenninger, and Stich affirmed 
 
 * Arloing, " Les Virus." Paris, 1891. 
 
_ 48 
 
 the diastasic nature of the putrid poison. The ven- 
 oms of serpents have for a long time been regarded 
 as soluble ferments. More recently the poison of 
 jequirity was considered an enzyme. This notion 
 has not, however, given any real advancement to 
 science, for the diastases themselves are perfectly 
 unknown to us in their chemical properties. 
 
 There is one exception, however: the diastases 
 are characterized by their determinate chemical 
 action on other substances. Pepsin, trypsin, and 
 papain dissolve albuminoid substances; ptyalin and 
 maltine break up starch into dextrine and glucose; 
 invertin inverts cane sugar; emulsin and saponin 
 decompose the glucosides. Now the only really 
 scientific way of ranking the microbian poisons among 
 the diastases would be to identify them from the 
 point of view of this determinate chemical action. It 
 is necessary to show that these poisons exercise a 
 hydrating action on a certain class of substances, and 
 that their toxic action proceeds precisely from this 
 chemical reaction. No serious attempt has been 
 made in this direction by the partisans of the diastasic 
 nature of the microbian poisons. They have ap- 
 pealed, it is true, to the great activity of these poisons 
 in very small doses. But from this point of view we 
 should have to rank among the diastases, hydrocyanic 
 acid, metilcarbylamine, the alkaloids such as nicotine 
 glucosides such as digitalin, salts of metals like cor- 
 rosive sublimate, metalloids like phosphorus and 
 fluorine. 
 
49 
 
 But there are still serious arguments against the 
 identification of the microbian poisons with the known 
 diastases. Fermi and others have studied the differ- 
 ent enzymes secreted by the microbes, and have found 
 that they have no appreciable toxic action.* 
 
 It follows that the toxic action of the bacteria is 
 not linked to their fermentative action; it does not 
 coincide with any known diastasic action. As for any 
 new diastasic action, we cannot deny it; at the same 
 time we cannot affirm it till we have found it. It is 
 evident that the toxic action of these poisons is re- 
 duced to some sort of chemical reaction, but we are 
 far from knowing this reaction. Consequently, to 
 say that the microbe poisons are diastases, is to speak 
 inexactly if we mean any diastases actually known, 
 and to juggle with words if we suppose a new diastase 
 with unknown action. On the other hand, we cannot 
 deny the numerous analogies between certain poisons 
 and the soluble ferments analogies which we shall 
 indicate later on. We must come down to the re- 
 searches of Brieger and Frankel before we find the 
 first serious attempt to determine the chemical nature 
 of the microbian poisons. This attempt has had 
 much success, although it was not made under the 
 conditions one could have desired. f 
 
 * Fermi: Die Hydrolytische Enzyme (Cent. f. Physiol- 
 ogic, 1891). 
 
 f Brieger and Frankel: Untersuchungen tiber Bacterien- 
 gifte (Berlin. Klin. Woch., 1890, Nos. n and 12). 
 
 4 OOO 
 
5 
 
 Brieger and Frankel first studied the diphtheritic 
 poison. They found that this poison behaves with 
 the different reagents like an albuminoid substance. 
 This poison is precipitated by the neutral salts in ex- 
 cess, such as the sulphate of ammonia and sulphate 
 of soda. It is not precipitated by sulphate of mag- 
 nesia. It is precipitated by absolute alcohol, and is 
 very soluble in water. By repeated precipitations by 
 alcohol, and dialysis (contrarily to Roux and Yersin, 
 Brieger and Frankel have seen that the poison does 
 not pass through the dialyzing membrane), the sub- 
 stance was obtained in a state of purity. This sub- 
 stance gave the following reactions: It is not pre- 
 cipitated by ebullition, sulphate of soda, common 
 salt, sulphate of magnesia, dilute nitric acid (even 
 with the aid of heat), or by acetate of lead. It is 
 precipitated by carbonic acid (in saturated solutions), 
 by the concentrated acids, by acetic acid and ferro- 
 cyanide of potassium, by phenol, by the organic acids 
 (soluble in excess), by sulphate of copper, by nitrate 
 of silver, by corrosive sublimate. It is laevogyrous. 
 From all these, and other reactions not mentioned, the 
 writers conclude that their toxic substance is exceed- 
 ingly like serum-albumin. They have even analyzed 
 this substance and found that it gives the elementary 
 composition of the albumins. 
 
 ~ Brieger and Frankel have also studied, but more 
 superficially, the other microbe poisons. They have 
 found, beside the toxic albumins, certain globulines, 
 
which are distinguished from albumins by their in- 
 solubility in distilled water. 
 
CHAPTER VI. 
 
 THE CHEMICAL NATURE OF THE BACTERIAL 
 POISONS (Continued). 
 
 SUMMARY. Criticism of the Work of Brieger and 
 Frdnkel Researches of Proskauer and Wasser- 
 mann The Microbe Poisons have Not yet been Pre- 
 pared in a State of Purity Similar Non-Microbian 
 Poisons The Work of Stillmarck Demonstration 
 of the Albuminoid Nature of the Diphtheritic Poison 
 Toxicohgical Differentiation of the Bacterial Poi- 
 sons Natural and Artificial Poisons: Their Char- 
 acters The Chemical Vaccines The Antitoxines or 
 Curative Substances Hypothesis as to the Chemical 
 Nature of the Poisons The Nucleo-albumins and 
 the Nucleints The Consequences of this Hypothesis. 
 
 We must dwell longer on the toxalbumins of 
 Brieger and Frankel, for the researches of these 
 writers have captivated the scientific world by their 
 apparent chemical rigorousness, and were followed 
 by the creation of toxalbumins, toxalbumoses, and 
 toxopeptones from every source. The conclusions of 
 Brieger and Frankel, nevertheless, call forth grave ob- 
 jections. And first, in order to prove that the diphther- 
 itic bacilli produce a special albumin, have they elimi- 
 nated the albuminoid substances from the bouillon ? 
 
53 
 
 Far from that, they have even added to the ordinary 
 bouillon which serves for the cultures the serum of 
 blood. How, then, can they believe that this serum- 
 albumin which they find again at the end of their 
 purifications is not the substance they introduced, 
 carrying down in its precipitations, as Panum had 
 already shown, the true microbian poison ? 
 
 This supposition becomes a certainty when we 
 regard the feeble toxicity of the pure substance of 
 Brieger and Frankel, compared with the far greater 
 toxicity of Roux and Yersin's impure product of their 
 calcic precipitate. In fact, Brieger and Frankel have 
 less right to say that the diphtheritic poison is a tox- 
 atbumin than one would have to affirm that it is phos- 
 phate of lime. 
 
 But phosphate of lime is a body well known, and 
 it is very easy to tell whether it contains impurities. 
 The albumins are much more difficult to analyze. 
 The proof of this is that Brieger and Frankel have 
 not been able to show us isolated the albumins of 
 broth and of serum, on the one hand, and the pretended 
 toxic albumin on the other. There is no doubt, then, 
 that in the experiments of Brieger and Frankel the 
 diphtheritic poison was always mixed with the albu- 
 mins of the culture media, and that the chemical 
 reactions described by them were those of the latter. 
 
 It may seem strange that these writers, when 
 studying the chemical nature of the microbian poi- 
 sons and finding them so closely associated with 
 
54 
 
 albumins, had not thought to first separate the albu- 
 mins from their cultures, leaving the microbes to 
 vegetate in media deprived of albuminoid substances. 
 But they had the preconceived idea that the bacterial 
 poisons could only be formed at the expense of the 
 albumins of the culture media. They give us no 
 proof of this notion, so widespread and yet so inexact, 
 as we shall show later on. 
 
 But even apart from the cultures in media de- 
 prived of albumins, there were many other means for 
 determining the chemical nature of the bacterial 
 poisons. 
 
 We might, for example, study what chemical 
 changes coincide with the heating of the filtered cul- 
 tures above 60 C. a temperature which destroys 
 the diphtheritic poison. We offer this suggestion 
 because Brieger and Frankel have made researches 
 which may seem of a like nature. They have studied 
 the non-toxic albumin produced by the emasculated 
 diphtheria-bacillus, and found it distinguished by sev- 
 eral constant characters from the albumin containing 
 the diphtheria poison. First, it is brown, while the 
 latter is white. It is soluble in dilute alcohol, and 
 combines with phenilhydrosine, while the toxic albu- 
 min does not combine with that body, but with chlor- 
 ide of benzoyl. It contains more carbon, less oxygen 
 and nitrogen, than the toxic albumin. As the virulent 
 bacilli are distinguished from the attenuated by the 
 energy of their assimilation, it is clear that this dis- 
 
55 
 
 tinction between the different albumins of their cul- 
 tures found by Brieger and Frankel must be of com- 
 plex origin, and does not permit us to judge as to the 
 nature of the diphtheritic poison., 
 
 We must conclude that, despite all the chemical 
 parade and apparatus of their work, Brieger and 
 Frankel have not succeeded in proving the legitimacy 
 of their conclusions that the microbian poisons are 
 albumins. Moreover, the same verdict has been 
 reached by the authors of another work, emanating 
 also from Koch's laboratory Wassermann and Pros- 
 kauer. 
 
 These writers have found in diphtheritic cultures 
 the two albumins of Brieger and Frankel the yellow 
 and the white. They have seen that the yellow albu- 
 min is especially preponderant in the cultures deprived 
 of toxicity; it is not itself toxic. The white albumin 
 was more abundant in the toxic cultures, and pos- 
 sesses a great degree of toxicity; by its chemical reac- 
 tions this toxic albumin corresponds to what Kuhne 
 calls albumose; it is precipitated by alcohol even when 
 dilute, by the concentrated acids, the salts of the 
 heavy metals, by potassium ferrocyanide, and by 
 acetic acid. It is not precipitated by ebullition, by 
 nitric acid, by basic acetate of lead, by sulphate and 
 chloride of sodium in excess. 
 
 But Wassermann and Proskauer are far from 
 being prepared to identify the diphtheritic poison 
 with this albumose. They remark, on the contrary, 
 
- 56 - 
 
 that the ordinary bouillon of culture always contains 
 certain albumoses by reason of the infusion of meat. 
 There does not exist, therefore, any serious argument 
 to favor the view that the toxic body which they have 
 isolated is not the same albumose of the bouillon with 
 which is found mingled the poison. This last idea is 
 confirmed by the fact that different portions of albu- 
 mose isolated from the same toxic culture differ in 
 their toxicity; they have, evidently, carried down vary- 
 ing quantities of the poison. The German writers 
 conclude that the albuminoid nature of the diphtheria 
 poison is possible, but that it has not been proved. 
 
 From all these researches we infer that the diph- 
 theria poison is closely associated with the albumin- 
 oid substances, and more particularly with albumose, 
 whose reactions it shares. 
 
 Just as in the case of the ptomaines, when chem- 
 ists sought for substances analogous to the alkaloids 
 of vegetal origin, the study of microbian toxalbumins 
 clearly discloses a parallel tendency in the efforts and 
 acquisitions of biological chemistry and of toxicology 
 /'. <?., towards the toxic vegetal and animal proteids. 
 In the seeds of different plants, in the poisons of ser- 
 pents, in the blood and bodies of certain animals, 
 were found certain substances of extraordinary viru- 
 lence not belonging to the class of alkaloids. We 
 cannot, unfortunately, enter here into the details re- 
 specting all these important researches. We will 
 content ourselves by mentioning here the writings of 
 
57 
 
 Warden and Waddel, of Martin and of Kobert, on 
 abrine; those of Weir Mitchell and Edward Reichart 
 and of Wolfenden on the poisons of serpents; those 
 of Kobert on the venomous spiders; those of Mosso 
 and of Kumahava on the poisonous fishes, etc.* 
 
 We shall here analyze only one of these memoirs 
 that of Stillmarck, composed under the direction of 
 Kobert, on Ricine. Stillmarck has found that it is 
 possible to extract from the seeds of the ricinus a 
 substance extraordinarily toxic, which presents all the 
 reactions of albuminoid bodies. It is not soluble in 
 alcohol, in ether, or in distilled water. It dissolves 
 readily in saline solutions. It is precipitated by ferro- 
 cyanide of potassium and acetic acid, by saturating 
 solutions containing it with the neutral salts, and by 
 ebullition. 
 
 Stillmarck supposes that this substance belongs 
 to the class of albumoses of Kuhne. Unlike these, 
 however, it is coagulated by heat. 
 
 As for the important question whether we can 
 identify the poison with this albuminoid substance, or 
 if the poison is simply mixed with the latter, Still- 
 marck advances serious arguments in support of the 
 first alternative. First, by boiling, ricine is coagulated 
 and rendered completely insoluble; at the same time, 
 its toxicity is completely abolished. Then again, pan- 
 
 *Vide the entire literature in Hulliburton's Text-Book 
 of Chemical Physiology and Pathology. London, 1891. 
 
- 58 - 
 
 creatic juice, which digests albuminoid substances, 
 destroys also the toxicity of ricine. 
 
 It is rendered very probable by these re- 
 searches, that there exist certain albuminoid bodies 
 of nature, yet undefined, which are very violent poi- 
 sons. We have recently shown that the diphtheria 
 poison must also be ranked in this category, for it is 
 destroyed, or at least modified, by the ferments of the 
 albuminoid bodies. 
 
 Now to what class of albuminoid bodies do the 
 microbian poisons belong? This question is the more 
 important because these poisons have an evident 
 kinship to a large class of vegetable and animal poi- 
 sons of which we have before spoken. 
 
 Unfortunately, the data which biological chem- 
 istry possesses respecting the albuminoid matters in 
 general are still very incomplete and indefinite. The 
 methods for their analysis are still very imperfect. It 
 is evident that under these conditions one cannot 
 rigorously determine the chemical nature of the 
 microbian poisons. We have already seen that even 
 the best studied of all these poisons, that of diph- 
 theria, has not yet been obtained in a state of purity,, 
 unmixed with the albumins of the culture media. 
 
 However, we possess for the microbian poisons a 
 precious and powerful means of analysis, namely, ex- 
 perimentation, which is based on the property of these 
 poisons to produce in animals certain phenomena 
 similar to those of the infectious diseases: intoxica- 
 tion and immunity. 
 
59 - 
 
 This means has enabled us to establish a funda- 
 mental distinction between two classes of microbian 
 poisons: the natural or primary poisons, and the 
 modified or artificial poisons. I have shown that in 
 cholera there exist two poisons totally different in 
 their physiological effects; the one produces the diar- 
 rhcea, while the other has a phlogistic action. These 
 poisons differ also by their resistance to heat; the 
 first is destroyed by heating it above 60 C., while the 
 second supports a temperature of 120 C. for several 
 hours. It is a tenable supposition that these two 
 poisons are closely associated together, and that the 
 second comes from the decomposition of the first.* 
 
 As for the diphtheritic poison, I have also found 
 that it is necessary to distinguish the secondary 
 poison which appears on the decomposition of the 
 primary poison by heat, by acids, by the soluble fer- 
 ments. 
 
 The attentive study of the other different poisons 
 enables us to classify them also in one or the other 
 of these two groups, which we^hall characterize more 
 in detail. 
 
 The natural poisons correspond to those which 
 Brieger and Frankel have called the toxalbumins, and 
 Klemperer the toxines. They are substances which 
 reproduce more or less exactly the symptoms of the 
 
 *Gamale!a: Experimental Researches on the Poisons 
 of Cholera (Arch, de Med. Experimentale, 1892, No. 2). 
 
6o 
 
 infectious disease. They are very unstable, and are 
 decomposed by a temperature above 60 C. The 
 animals which are refractory to the disease produced 
 by their microbe, are also refractory to the primary 
 poison of this microbe. 
 
 The modified poisons, which are also named 
 proteins (Buchner, Klemperer), do not reproduce the 
 typical phenomena of the microbian disease. They 
 cause hypothermia or fever, according to the dose, 
 inflammation more or less intense at the point of in- 
 oculation, and symptoms more or less pronounced of 
 cachexia. They have the remarkable property of 
 exciting tuberculous animals to general and local re- 
 action. These poisons resist boiling. They are pre- 
 cipitated by alcohol. Panum's poison probably be- 
 longs to this class of substances. These poisons have 
 nothing to do with the production of immunity. In 
 certains cases, on the contrary, they have an opposite 
 influence, predisposing the economy to the invasion 
 of microbes. The vaccinated animals are not refrac- 
 tory to these artificial poisons. By the side of these 
 primary and secondary poisons, experimental analysis 
 has revealed still a third and a distinct order of 
 substances. These are the chemical vaccines the 
 microbian products which confer on animals immu- 
 nity against infection by the living microbe. In 
 certain cases the chemical vaccination may be ob- 
 tained by means of the primary poison as, for in- 
 stance, in the vaccinations against the septic vibrio, 
 
6i 
 
 or against the streptococcus of pneumonia. But in 
 many other cases the primary poisons do not confer 
 immunity, but lead, even in small doses, to cachexia. 
 Such is the case with regard to tetanus and diphtheria. 
 But here also a chemical vaccination is possible by 
 means of certain artifices. Thus, for instance, we 
 prepare the vaccines by heating the primary poisons 
 above their critical point, to 70 or 80 C. We may 
 suppose that this heating sets free the vaccinal sub- 
 stance of the primary poison. This method succeeds 
 for pneumonia, and sometimes for tetanus and diph- 
 theria. In some diseases chemical vaccination is easier; 
 the microbian products vaccinate, even after having 
 been subjected to a temperature of 100 and even 
 120 C. This is the case with cholera and the vibri- 
 onic septicaemia, which I was the first to discover. 
 
 In certain cases the vaccinant products have 
 been quite isolated from the primary or modified 
 poisons. I have found that against cholera and 
 against the avicide vibrio we may vaccinate with the 
 volatile products of the cultures. As for the proteus 
 vulgaris, it has been seen that we can vaccinate by 
 the different ptomaines, such as neuridine, muscarine, 
 etc. Thus we see that the chemical vaccines are very 
 varied. We may say, in general, that they are more 
 stable than the corresponding poisons. I have re 
 garded them as a decomposition-product of the poi- 
 sons; in any event, they have a variable chemical, 
 nature, being decomposed by ebullition. 
 
62 
 
 There exist, lastly, substances still more com- 
 plex, which are found only in the organism of re- 
 fractory animals after an injection of the poisons. 
 These substances, which are combinations of poisons 
 with the animal substances, or possibly chemical vac- 
 cines quite pure, have the property not only of vac- 
 cinating the animals against an infection to come, 
 but of curing the disease when declared.* The nature 
 of these immunizing and curative substances is com- 
 pletely unknown. 
 
 If we wish now to sum up all this long study of 
 the chemical nature of the microbian poisons, we 
 arrive at the following conclusions: 
 
 Among the products of the microbes, there are 
 four classes of substances: First, the organic bases 
 (which, if they do not exist essentially as ptomaines, 
 assume that form under the different processes of 
 analysis); second, certain substances of albuminoid 
 nature, or primitive poisons, easily decomposible at a 
 temperature above 60 C.; third, still other albumin- 
 oid substances, which are more stable the modified 
 poisons; fourth, a class of substances which can only 
 be defined by their physiological action, the chemical 
 vaccines these are associated with one of the three 
 preceding classes of microbian products. 
 
 Despite all this diversity of the microbian prod- 
 uts, and all the apparent complexity of their relations, 
 
 *GamaleIa: Immunisation (Gaz. Hebd., 1891, No. 47). 
 
-63 - 
 
 we may frame a hypothesis to explain their chemical 
 nature. 
 
 Biological chemistry has taught us that there is a 
 particular class of substances more complicated than 
 the ordinary albuminoid substances, extraordinarily 
 unstable, decomposing even by prolonged contact 
 with alcohol and by heat above 60 C., giving rise by 
 their decomposition to the formation of other albu- 
 minoid substances more stable, as well as to the dif- 
 ferent ptomaines and leucomaines. These are the 
 nucleo-albumins or vitellines, constituent parts of all 
 animal and vegetable cells. I have made the hypo- 
 thesis that the natural poisons of the bacteria are 
 precisely such nucleo-albumins, which give rise to 
 nucleines or modified poisons by their decomposition, 
 and to ptomaines by a more profound decomposition. 
 The principal chemical character of the nucleo-albu- 
 mins is the great quantity of phosphorus which they 
 contain. In the microbian poisons, chemists have 
 already found much phosphorus. Such is the case in 
 regard to the poisons of cholera (Petri), the avicide 
 vibrio (Wolkow), and Koch's tuberculin. But all these 
 acquisitions of organic chemistry have no decisive 
 value, for no one of these poisons has been obtained 
 in a state of purity all have been mixed with other 
 substances derived from the culture media. Till the 
 determination of phosphorus shall have been made 
 for a perfectly pure microbian poison, our idea will 
 remain only an hypothesis. But it may be verified 
 
6 4 
 
 upon other characters of the nucleo-albumins char- 
 acters which are less decisive from a chemical point 
 of view, but much more interesting from the stand- 
 point of microbian toxicology. 
 
 And, first, there arises the fundamental question: 
 Are the microbian poisons the products of decom- 
 position of the nutritive media of the bacteria, as cur- 
 rent opinion has it, or are they the constituent parts- 
 of the bodies of the microbes, as they ought to be if 
 they were nucleo-albumins? This important question 
 concerning the origin of the microbian poisons wilt 
 be examined in the next chapter. 
 
CHAPTER VII. 
 
 THE ORIGIN OF THE MICROBIAN POISONS. 
 
 SUMMARY. Preconceived Idea that the Poisons Can 
 Only Come from the Decomposition of Albumin- 
 oid Bodies Experiments Already Old which Over- 
 throw this Idea Researches of Polotebnoff, Popoff, 
 Bergmann, Schuller Numerous Researches Made 
 at the Laboratory of Pachoutine Recent Labor of 
 Guinochet The Microbian Poisons Are Not Prod- 
 ucts of Decomposition, but the Result of Synthesis 
 Are They Secretions ? Arguments in Favor of this 
 Idea Arguments which Combat It The Poisons 
 are Intimately Linked to the Bodies of the Bacteria 
 
 The Experiments of Cantani and of the Author 
 
 The Researches of Buchner on the Proteins 
 Criticism of these Researches All the Microbe 
 Poisons Come from the Bodies of Bacteria Utility 
 of this Source for the Microbes. 
 
 In all the history of the researches on the 
 microbian poisons, the idea was generally admitted 
 that these poisons take their origin in the course of 
 the decomposition of albuminoid matters. This idea 
 is prevalent at the present time. Nevertheless, there 
 
 exist in science many facts which completely over- 
 
 5 ooo 
 
66 
 
 throw it. As early as the epoch of the study of the 
 putrid poison, chemists were seeking to eliminate the 
 albuminoid substances from the matters undergoing 
 putrefaction, in order to render easier the extraction 
 of the putrid poison. Thus, for instance, Bergmann 
 and Schmiedeberg had employed brewer's yeast in 
 their famous experiments on sepsine. 
 
 Polotebnoff was the first to employ an entirely 
 mineral medium and allow it to putrefy. It was 
 Pasteur's liquid, composed of ammonia and of dif- 
 ferent neutral salts. Polotebnoff found that the 
 liquid when putrefied is not more toxic than the fresh 
 liquid. Popoff, who repeated the experiments of 
 Polotebnoff, has shown that these conclusions are 
 not exact, and that Pasteur's liquid when putrefied 
 produces absolutely the same symptoms of intoxica- 
 tion as the putrefied albuminoid matters. But Popoff 
 did not kill the microbes in his liquid, and he sup- 
 posed that it was precisely the microbes which pro- 
 voked this septic intoxication. The same experiments 
 were made by Hugo von Brehm. Bergmann has 
 given to the question a decisive advancement. He 
 has shown that Pasteur's' liquid when putrefied pre- 
 serves all its septic action, even after having been 
 boiled. Anders has also found that neither boiling 
 nor filtration through clay destroys the septic action 
 of this liquid. 
 
 Thus was established this principle of the highest 
 importance: That the septic poison is the result of 
 
- 67 - 
 
 the synthetical action of the microbes, and not of the 
 decomposition of the putrid matters. 
 
 The same results were obtained by Schuller, who 
 operated with the liquid of F. Cohn, which contains 
 no sugar, and whose sole organic compound is am- 
 monium tartrate. 
 
 These experiments have since been often repeat- 
 ed with the same positive results. These researches 
 have been chiefly made at the laboratory of Pachou- 
 tine. We must mention especially the theses of Bott- 
 charoff and of Cosorotoff emanating from this labora- 
 tory. All these writers, as well as numerous others 
 who have studied the details of putrid intoxication as, 
 for instance, the fever and the gaseous exchanges- 
 have uniformly arrived at this result: that the putrid 
 poison is formed in the absence of every albuminoid 
 body, and that it is, consequently, the result of the 
 synthetic activity of the microbes. Note also that 
 these writers have .confirmed in general the datum 
 of Panum as to the great toxicity of the products 
 precipitated by alcohol. Nevertheless, contrarily to 
 Panum, the alcoholic extract had often itself also a 
 certain septic activity. 
 
 Unfortunately, all these experimental researches 
 were carried on, not with pure cultures of bacteria, 
 but with cultures containing the microbes of putre- 
 faction in indefinite quantity. They were carried on 
 from the point of view of experimental pathology, 
 and bacteriology, which has mostly confined itself to 
 
68 
 
 pure cultures of microbes, has rather ignored the im- 
 portant results established by these labors. 
 
 Of much more importance are the researches of 
 Guinochet on the diphtheritic poison. Guinochet has 
 found that the diphtheritic bacillus, which is easily 
 cultivated in normal filtered human urine containing 
 not a trace of albumin, produces its poison in that 
 medium quite as energetically as in meat broth.* 
 
 Thus we find that there is little ground for the pre- 
 conceived idea, which has lasted so long, that the 
 pathogenic microbes fabricate their poisons by a 
 special decomposition of albuminoid matters. This 
 notion, demonstrated false, must give place to the 
 more just conception, viz., that the poisons are the 
 synthetic products of the microbes. 
 
 Independently of the theoretical importance of 
 this conclusion of Guinochet, we must also indicate 
 its practical side. We shall be able henceforth to 
 study the formation of the micrabian poisons in con- 
 ditions more simple than those which were chosen 
 by Brieger and Frankel and their successors, in a 
 medium totally deprived of albumins, f 
 
 The question now presents itself: What is the 
 role of these substances ? What place do they oc- 
 
 * Guinochet: Labors of the Laboratory of Straus, at the 
 Ecole de Medecine, Paris, France. 
 
 fit only remains to find a culture medium favorable to 
 the development of the microbes, for the mineral media 
 (liquids of Pasteur, Noegeli, and Cohn) are not suitable for 
 
-6 9 - 
 
 cupy in the life of the microbes ? For the definition 
 of animal poisons depends for its significance upon 
 the relation of these poisons to the higher organisms, 
 and not to microbe-biology. 
 
 There were already a certain number of syn- 
 thetic and specific products known to microbiology, 
 whose signification was sufficiently clear. These 
 were the soluble ferments, or. diastases, which the 
 microbes secrete to render assimilable the aliments 
 which surround them. The yeast-plant secretes inver- 
 tine, which decomposes cane sugar, not directly utiliz- 
 able, into dextrine and levulose. The bacillus of lactic 
 acid secretes the lab-ferment (which serves to coagu- 
 late the casein of milk), and casease (which dissolves 
 and peptonizes it). 
 
 It was natural to think of the similarity of these 
 microbian manifestations to the empoisonments pro- 
 duced by the pathogenic microbes. One might readily 
 believe that the poisonous secretions serve also to 
 prepare the animal soil for the microbian life, whether 
 by rendering the animal humors assimilable for the 
 microbes, or by abolishing the vital resistance of the 
 
 the pathogenic bacteria. I have found that they grow well 
 in the following medium: 
 
 Water 1000 
 
 Liebig's extract 5 
 
 Glycerin 4 
 
 Common salt 5 
 
 Liebig's extract is totally exempt from albuminoid 
 matters. 
 
70 
 
 animals and thus rendering the field clear for the oc- 
 cupancy of the microbes. 
 
 This notion soon found a certain support in 
 experimentation. It has been observed that the 
 diphtheritic and tetanus poisons behave towards the 
 different reagents precisely like the soluble ferments. 
 It has also been noted that different substances the 
 soluble ferments of papain; such microbian products 
 as the poison of the bacillus prodigiosus; the poisons 
 of plants, like abrine also possess the property of 
 rendering the animals in which we inject them in min- 
 imum doses predisposed to the pullulation of microbes 
 which are not pathogenic to the same animals in the 
 normal state.* 
 
 Nevertheless this comparison of the poisons to 
 the secretions accounts but superficially for the facts. 
 
 First, there exist certain pathogenic bacteria 
 which produce such diseases (of the highest import- 
 ance) as tuberculosis, cholera, and charbon, where 
 we do not find any toxic secretions. 
 
 Then again, even the production of poisons in 
 diphtheria and tetanus has a very superficial corre- 
 spondence to a secretion. In fact, the quantity of 
 these poisons in the culture liquids does not grow 
 proportionately -to the development of the bacteria 
 themselves. On the contrary, an attentive observa- 
 tion reveals the following facts: 
 
 *See GamaleJa "On the Reproduction of Cholera in 
 the Hare" (Congress of Berlin, 1890). 
 
At the very first, and at the most active moment 
 of the life of the microbes, the liquid of culture is acid 
 and devoid of all toxic power. Later, when the bac- 
 teria have ceased to multiply and are deposited at 
 the bottom of the culture-flask, the liquid becomes 
 alkaline and more and more toxic. Its toxicity aug- 
 ments, in a certain measure, progressively with the 
 duration of the sojourn of the bacteria in their alka- 
 line liquid of culture. 
 
 The explanation of this fact is very simple. The 
 poison is contained inside the bodies of the bacteria, 
 -and is extracted but slowly by the alkaline liquid in 
 which they are macerated. 
 
 This hypothesis as to the origin of all the 
 microbe poisons from the bodies of the bacteria has 
 been already proposed by Cantani a propos of the 
 cholera poison. Cantani supposed that in the cholera 
 cultures it was the bacilli themselves which were 
 toxic, like certain fungi which cause poisoning when 
 injected. He did not prove his hypothesis, which long 
 remained without support. This was six years ago, 
 .and Cantani's article "Die Giftigkeit der Cholera- 
 bacillen " was published in the Deutsche Medicinische 
 Wochenschrift in 1886. Since then I have shown 
 that the cadavers of divers bacteria remain very 
 toxic even after having been exposed to a heat of 
 100 to 120 C. 
 
 Later, I showed that we can prepare from the 
 cadavers of the cholera vibrio and of the avicide vibrio 
 
72 
 
 very toxic extracts which possess at the same same 
 vaccinal properties.* 
 
 At the same time that this study of the general 
 action of the bacterial cadavers was being pursued, 
 other experimenters were investigating their local 
 effects. After Gravitz and Wyssokowitch had shown 
 that the cadavers of certain bacteria have the prop- 
 erty of producing suppuration in animals, Buchner 
 made of this question a special study. By boiling 
 with dilute alkalies, he succeeded in extracting from 
 the bodies of different bacteria, such as the typhoid 
 and the pyocyanic bacillus, substances in general of 
 little toxicity, but possessing the property of producing^ 
 by subcutaneous injection an exudative inflamma- 
 tion. With doses still more feeble, these poisons 
 provoke only an emigration of leucocytes (positive 
 chimiotaxy). 
 
 By their chemical properties these substances 
 correspond quite well to those which Rencki had 
 described under the names of myco-protein and antraco- 
 protein. Buchner considered them as alkali-albumins 
 or proteins, and under that name they have since 
 been studied. Their most curious property has been 
 set forth by Koch. Already in 1889 I had indicated 
 that tuberculous guinea-pigs have an extreme sus- 
 ceptibility towards the vibrio poison. Koch has sue- 
 
 *GamaleIa, in Ann. de 1" Instit. Pasteur, 1888, No. 5; 
 also 1889, No. 10; also in Compt. Rend, de la Soc. de 
 Biologic, Nov. 30, 1890. 
 
T 3 
 / O 
 
 ceeded in extracting from the tubercle bacilli, by 
 heating with glycerin, a substance called tuberculin, 
 which proves to be extremely active in tuberculous 
 subjects. It provokes in them the phenomena, so 
 often described, of general and local reaction. This 
 reaction of the tuberculous to tuberculin has long 
 been wrongly considered as a characteristic effect 
 of the latter. But latterly it has been shown that, 
 besides the avicide vibrio and the bacillus tuber- 
 culosus, many other bacteria contain a substance hav- 
 ing a similar property. Buchner and Rohmer have 
 given the following directions for the extraction of 
 these substances: Dry the bacteria after separating 
 them from their culture media; boil them several 
 hours in alkaline water, and leave the decoction to 
 macerate for several days at the temperature of 37 
 C; then filter the liquid, and precipitate the active 
 substance by acetic acid. Thus we see that, with the 
 proteins of Buchner, the poisons extracted from the 
 bodies of bacteria have acquired the right of place in 
 bacteriology. But we must make some important 
 reservations in regard to this conception of the pro- 
 teins. 
 
 First, Buchner and the chemists who followed 
 him established a fundamental distinction between 
 the proteins and the toxalbumins or toxines for the 
 latter come from the secretions of the bacteria, the 
 former only from their bodies. The proteins exercise 
 a positive chimiotaxy, the toxines negative chimio- 
 
74 
 
 taxy. (We shall return shortly to the role of micro- 
 bian poisons in inflammation.) As to the source of 
 these poisons, we have already seen that there are 
 reasons to suppose that the toxalbumins are also con- 
 tained in the bacteria. If Buchner has not succeeded 
 in extracting from the bodies of microbes aught but 
 the proteins, this is evidently due to his crude method 
 of decoction, which surely destroys the toxalbumins. 
 We have shown that in cholera one may obtain by 
 extraction at a temperature of 55 to 60 C. another 
 poison than that which is extracted by ebullition. 
 Likewise, in respect to tuberculosis, if instead of 
 Koch's process you employ Weyl's more efficacious 
 method of extraction by alkaline water at low tern- 
 peratures, you obtain a poison more active than 
 tuberculin, and endowed with other physiological and 
 chemical properties. Now it is more simple to sup- 
 pose that all the poisons of the microbes come from 
 their bodies; but that the extraction at low tempera- 
 tures gives us the toxalbumins or primary poisons, and 
 that ebullition gives the proteins or modified poisons. 
 Moreover, in coming back to the point of de- 
 parture of the researches of Buchner to the suppura- 
 tion produced by the microbian cadavers it is proper 
 to note that this savant has completely failed in his 
 attempt to find the pyogenic substances. The pro- 
 teins which he has isolated produce an exudative 
 inflammation, and not suppuration. Buchner has 
 found the likeness of suppuration in the emigration 
 
75 
 
 -of the leucocytes, in the positive chimiotaxy provoked 
 by his proteins. But this positive chimiotaxy not only 
 has not the significance of suppuration, but is 
 not even inflammation. It is nothing but hyper- 
 semia. We see it when we introduce very small doses 
 of proteins (in capillary tubes) under the skin of 
 animals. When we augment the doses injected, we 
 observe (as we have seen) the production of a serous 
 exudative inflammation without leucocytes. So our 
 positive chimiotaxy becomes, by the augmentation of 
 the doses, negative chimiotaxy, and cannot serve to 
 distinguish the two classes of poisons. But suppura- 
 tion is not obtained with the proteins. 
 
 It is very probable that the failure of Buchner 
 to extract pyogenic substances from the microbian 
 bodies was due to his faulty process of extraction, 
 and that the pyogenic substances belong to the cate- 
 gory of the more fragile (the primary) poisons. As to 
 the chemical nature of the substances extracted by 
 Buchner from the bodies of the bacilli, their identity 
 with the class of alkali-albumins or proteins cannot 
 longer be admitted. Buchner had found that these 
 substances present the reaction of albuminoids, and 
 that they are, besides, precipitated by neutralizing with 
 acetic acid their alkaline solutions. But this latter re- 
 action is an artificial one, produced by reason of their 
 extraction by alkalies from the bodies of bacilli. When 
 Buchner later employed for this extraction neutral 
 water, he no longer observed precipitation by the 
 
76 
 
 acids. If we still continue to call these substances 
 alkali-albumins, it is by an abuse of language. Like- 
 wise, Koch's tuberculin, which belongs by all its 
 physiological properties to the same class as Buchner's 
 proteins, but which is extracted by means of a neutral 
 solvent, does not present the characteristic reactions 
 of the alkali-albumins. It is evident, consequently, 
 that the substances of Buchner, or our modified 
 poisons, are not alkali-albumins. We have already 
 stated in the preceding chapter that we believe them 
 to be nucleines. As for tuberculin especially, our 
 hypothesis is found confirmed by the richness of this 
 substance in phosphorus, which we find even after 
 having freed from phosphoric salts the bouillon in 
 which the bacillus tuberculosis has vegetated. 
 
 Thus the modern researches on the origin of mi- 
 crobian poisons lead us to the same results as the study 
 of their chemical properties. The microbian poisons 
 may be divided into primary and secondary. They 
 are the results of the creative synthesis of the 
 bacteria, and come from their bodies. Thus is again 
 confirmed our hypothesis respecting their chemical 
 nature which identifies them with the constituent 
 parts of the nuclei of cells the vitellines and the 
 nucleines. 
 
 The toxicity of the microbian bodies may have 
 also a teleological signification, explicable from the 
 point of view of the struggle for existence and of 
 natural selection; for the toxicity of the bodies of 
 
77 
 
 bacteria may be useful to them as a means of pre- 
 serving them against aggressors which would devour 
 them. 
 
CHAPTER VIII. 
 
 ACTION OF THE BACTERIAL POISONS ON THE 
 
 ANIMAL ORGANISM ACCUSTOMANCE 
 
 AND IMMUNITY. 
 
 SUMMARY. Insufficiency of our Knowledge of the Mode 
 of Action of the Bacterial Poisons on the Animal 
 Organism Local Action of the Modified Poisons 
 Selective Action of Tuberculin and Malleine De- 
 struction of Poisons in the Bodies of Refractory 
 Animals Explanations of the Different Contradic- 
 tions Re'sume' and Conclusions on the General 
 Toxicity of Microbes. 
 
 As the microbian poisons have not been the 
 object of systematic researches in their aggregate, cer- 
 tain important questions concerning their history are 
 still undecided. The mode of action of the poisons 
 on the organism is one of these questions. Even in 
 the case of the best known of the microbian poisons, 
 we do not yet know on what elements they act pref- 
 erably, whether on the nerve-cells or muscles, on 
 the heart or capillary vessels, or even on the blood. 
 
 Most of these poisons produce a local lesion at 
 the place of injection. This local lesion may go on 
 from a serous or sero-sanguinolent exudation to 
 necrosis. This phlogistic action belongs to the- 
 
79 
 
 majority of the modified poisons. It is, too, a little 
 singular that when injected into the blood, these same 
 poisons have a marked antiphlogistic action. The 
 intravenous injection of these poisons prevents the 
 appearance of inflammation, even when solicited by 
 very energetic agents, such as the application of cro- 
 ton oil. Bouchard has endeavored to explain this 
 antiphlogistic action by the paralysis of the vaso- 
 motor centres; but this explanation is not fully 
 demonstrated, and still remains contested (Samuel).* 
 
 When injected under the skin, but in very small 
 doses, the same modified poisons provoke inflamma- 
 tion, not at the place of their introduction, but around 
 foci of pre-existent lesions. The best known examples 
 of this elective action are the local reactions provoked 
 by tuberculinf in the tuberculous, and by malleinej in 
 horses affected with glanders. 
 
 The attempt has seriously been made to explain 
 this elective inflammation by the positive chimiotaxy 
 
 *Charrin and Gamaleia : On Inflammation (C. R. de 
 la Societ6 de Biologic, July 5, 1890). Gamaleia : On the 
 Local Lesion in Microbian Diseases (Arch, de Med. Exper., 
 1891, No. 2). 
 
 f Koch, Deutsche Medicinische Wochen., 1890, No. 46. 
 Gamaleia: On the Treatment of Tuberculosis by Koch's 
 Method (Arch, de Med. Experimentale, 1891, No. 2). 
 
 JHelmann: On the Clinical and Experimental Diag- 
 nosis of Glanders (Messager de la Science Veterinaire Pub- 
 lique, 1891, No. 4). 
 
8o 
 
 excited by tuberculin when freed from glycerin. This 
 explanation is at the best a strange one. 
 
 How can the positive chimiotactic power of 
 tuberculin, i.e., the action which it may exercise in 
 certain conditions on the leucocytes, explain the exu- 
 dative inflammation produced by tuberculin, not at 
 the spot of inoculation, but at the place of election ? 
 This impossible comparison can only be understood 
 by calling to mind the confusion which exists in cer- 
 tain microscopists with regard to the difference be- 
 tween inflammation and the emigration of leucocytes, 
 the latter being only an element more or less constant 
 of inflammation, and indeed oftener found apart 
 from it.* 
 
 The explanation of the elective action of tuber- 
 culin is not, however, so difficult, and it has already 
 been sketched by Koch. The acute inflammation is 
 in general the consequence of a lesion or rapid de- 
 struction of the cells and tissues of the animal. When 
 injected in sufficient concentration, the modified 
 poisons induce cellular lesions and exudative inflam- 
 mation at the place of their injection. Appropriately 
 diluted, they are inoffensive to the normal cells of 
 the economy, and affect only the tissues already dis- 
 eased, around which is then produced the inflamma- 
 tory process. This explanation agrees well with the 
 
 *This confusion has even been made the basis of a new 
 theory of inflammation. 
 
8i 
 
 property of other substances besides tuberculin to 
 provoke reaction in the tuberculous, and with the 
 property of tuberculin of provoking also reaction 
 around certain lesions not tuberculous. 
 
 The relations between the microbian poisons and 
 the animal economy have been chiefly studied from 
 the point of view of immunity. When vaccination by 
 means of the soluble products of bacteria was first 
 demonstrated, the very natural idea arose that this 
 immunity was acquired by the habituation of the ani- 
 mals to the poison. According to the different hypoth- 
 eses respecting immunity, this habituation was sup- 
 posed to affect either the entire animal economy, or 
 only certain of its cells. This idea was, moreover, con- 
 firmed by the facts connected with one of the first 
 chemical vaccinations known. Beumer noted that he 
 could easily habituate mice to progressively increas- 
 ing doses of the sterilized cultures of the typhoid bacil- 
 lus. These mice, having become accustomed in this 
 way to the poison, were able to sustain without injury 
 the inoculation of large quantities of the living bacilli 
 a procedure which proved speedily fatal to mice not 
 vaccinated.* Similar facts have been noted by Foa 
 and Bonome in the chemical vaccination of hares 
 against the proteus vulgaris.f 
 
 * Beumer: Der derzeitige Shandpunkt der Schutzim- 
 pfungen, pp. 4-6. Wiesbaden, 1887. 
 
 f Foa and Bonome: Ueber Schutzimpfungen (Zeitschrift 
 
 f. Hygiene, t. v, p. 415). 
 6 ooo 
 
82 
 
 But in my own researches on vaccination against 
 the avicide vibrio found by me, I have met with 
 facts which overthrow this theory of habituation. 
 The cultures of the avicide vibrio, sterilized at 
 120 C. and very toxic for guinea-pigs, easily vac- 
 cinate them against infection by the living vibrio. 
 But after being once vaccinated the guinea-pigs 
 do not become more resistant to the toxic action 
 of the vaccine /. <?., to massive doses than the non- 
 vaccinated. The fever which appeared after the 
 injection of small doses was in them identical with 
 that of the control-animals. The hypothermia fol- 
 lowing larger doses had the same characters. The 
 exudative inflammation which the vaccine provokes 
 at the point of inoculation was not modified in conse- 
 quence of the vaccination of the guinea pigs. The 
 phenomena of positive or negative chimiotaxy de- 
 pended on the dose of vaccine introduced, but not on 
 the immunity or non-immunity of the guinea-pigs. 
 In short, the behavior of the guinea-pigs toward the 
 poison of the chemical vaccine was in no way 
 changed by the vaccination. Moreover, we have 
 found that the volatile products of the non-toxic 
 cultures can also vaccinate. We have found this 
 same failure to undergo habituation in animals 
 subjected to cholera vaccine,* to pyocyanic vac- 
 
 *The facts which we have found have been confirmed 
 by many observers. See especially Hernandez, "Chemical 
 Vaccinations," in C. R. de la Soc. de Biologic, July 4, 1891; 
 
83 - 
 
 cination (Charrin and I), to hog-cholera vaccine 
 (Selander).f 
 
 All these facts show that the chemical vaccina- 
 tion of animals does not depend on their becoming 
 accustomed to the vaccine. They may be explained 
 by the hypothesis that in our vaccine are mingled the 
 poison, and the vaccinant substance distinct from the 
 poison. This hypothesis of the distinction between 
 the toxic substance and the vaccinant substance, 
 formulated first by Bouchard, J may account for the 
 mechanism of vaccination, but it is manifestly insuffi- 
 cient, for it does not explain the positive cases of re- 
 sistance to the poisons by the vaccinated animals in 
 the experiments of Beumer, Foa, and Bonome, and 
 others of whom we shall speak presently. 
 
 The complete explanation of this difficulty can 
 only be obtained by the aid of the notion of the 
 plurality of the microbian poisons. We have estab- 
 lished the distinction between the natural or primary 
 poisons of the microbes, and their artificial or modi- 
 
 and Metschnikoff and Roudenko, " Researches on Habitua- 
 tion to the Microbian Products " (Ann. de 1'Instit. Pasteur, 
 1891, No. 9). 
 
 f Gamaleia: On Cholera Vaccination (C. R. de la Soc. 
 de Biologic, Nov. 30, 1891). Charrin and Gamaleia: Vacci- 
 nation et Accoutumance (C. R. de la Soc. de Biologic, July 
 5, 1890). Selander: Study of Hog-Cholera (Ann. de Tlnst. 
 Pasteur, 1890, p. 545). 
 
 t Bouchard: Therapeutique des Malades Infectieuses, 
 p. 137. Paris, 1889. 
 
_ 8 4 - 
 
 fied poisons.* With this distinction, all the foregoing 
 contradictions are cleared up. If our animals vac- 
 cinated against the infection did not become refrac- 
 tory to the poisons contained in the chemical vac- 
 cines, this was owing to the fact that the latter 
 poisons are artificial products and have nothing to do 
 with the infection and the natural poisons of the bac- 
 teria. 
 
 In fact, towards these natural poisons the vac- 
 cinated animals become quite as refractory as towards* 
 the microbes themselves. This last fact was first 
 established by the researches of Behring and Kitasata 
 in reference to tetanus and diphtheria, f G. and F. 
 KlempererJ have noticed the same resistance to the 
 pneumonic poison on the part of animals vaccinated 
 against the streptococcus lanceolatus of pneumonia. In 
 cholera, lastly, we have seen that vaccinated dogs 
 become refractory to the natural poison of cholera 
 and not to the modified poison. Quite recently, also, 
 
 *GamaleIa: On the Diarrhoeic Action of the Cultures of 
 Cholera (C. R. de 1'Acad. des Sciences, March 24, 1890). 
 Gamalela: On the Two Poisons of Cholera (Bulletin Medical,. 
 April, 1890). 
 
 f Behring and Kitasato: Facts as to Immunity from, 
 Tetanus and Diphtheria (Deutsche Medic. Wochensch., 1890^ 
 No. 49)- 
 
 $G. and F. Klemperer: Immunity against Pneumo- 
 coccus Infection (Berlin. Klin. Wochensch., 1891, Nos. 34 
 and 35). 
 
- 85 
 
 in respect to typhoid fever, the researches of Beumer 
 and Peiper on the resistance of vaccinated animals to 
 the poison have been confirmed by those of Brieger, 
 Kitasato, and Wassermann.* 
 
 We may consequently regard it as a very general 
 fact that animals refractory to infection by a microbe 
 also resist intoxication by the natural poison of this 
 microbe. This principle finds verification, not only 
 in reference to artificial immunity conferred by vac- 
 cination, but also to natural immunity. Thus, for 
 instance, rats which are refractory to the diphtheritic 
 infection, support without difficulty, according to 
 Roux and Yersin, the injection of enormous doses of 
 the poison of diphtheria. 
 
 We may conclude that immunity is linked to the 
 resistance of animals to the microbian poisons. 
 
 To what is this resistance due? We have already 
 seen that it is not the result of accustomance, for by 
 a chemical vaccine which does not contain any natural 
 poison we vaccinate against a natural poison. 
 
 Behring and Kitasato have shown that in respect 
 to tetanus and diphtheria, acquired immunity and re- 
 sistance to the poison are linked to the power of the 
 serum of the vaccinated animals to destroy even in 
 vitro the corresponding poison. This remarkable dis- 
 covery has been since then often confirmed in its ap- 
 
 * Brieger, Kitasato, and Wassermann, Zeitschrift f. 
 Hygiene, t. xii, p. 137. 
 
86 
 
 plication to diphtheria and tetanus, and also to other 
 poisons.* 
 
 At the same time, this antitoxic property of the 
 serum cannot be the cause of the resistance of ani- 
 mals to the microbian poisons; it is rather the con- 
 sequence, for animals naturally refractory do not 
 ordinarily possess this antitoxic property of the 
 serum they do not acquire it until after having de- 
 stroyed the poisons in their bodies. f The immunity 
 cannot therefore depend on the antitoxic power of 
 the serum; it can only be characterized by the pos- 
 sibility of acquiring this antitoxic power after the in- 
 troduction of the poison. 
 
 But the question as to the nature of immunity is 
 foreign to our book, and we cannot enter into expla- 
 nations respecting its physiological mechanism. 
 
 To sum up, briefly, all that we know as to the gen- 
 eral toxicology of the microbes, we shall see that our 
 hypothesis as to the chemical nature of the bacterial 
 poisons, in absence of a direct demonstration which 
 can only be made with the poisons isolated in a 
 state of purity, is found confirmed by many indirect 
 proofs. 
 
 That the microbian poisons belong to the class" of 
 nucleo-albumins, is in agreement with their extreme 
 
 *Ehrlich: Experimentelle Untersuchungen tiber Immu- 
 nitat (Deutsche Medic. Woch., 1891, Nos. 32 and 44). 
 
 fGamaleia: L'immunisation (Gaz. Hebdom. de Med. 
 et de Chir., Nov. 21, 1891). 
 
8 7 
 
 fragility, and the facility with which they give rise to 
 the modified poisons on the one hand, to the pto- 
 maines on the other. This idea is also confirmed by 
 what we know about the source of the poisons of the 
 microbian bodies, which are of nuclear origin. It is, 
 lastly, compatible with the plurality of poisons asso- 
 ciated together and produced by the same bacteria a 
 plurality postulated by the physiological study of 
 these poisons. 
 
 We shall speak only of the principal facts which 
 confirm our hypothesis. 
 
Third Part: The Special Toxicology of 
 the Microbes. 
 
 CHAPTER IX. 
 
 THE POISONS OF TETANUS. 
 
 SUMMARY. Researches of Brieger, Kitasato and Weyl, 
 Knud Faber, Tizzoni and Cattani, Brieger and 
 Frdnkel, Vaillard and Vincent and Kitasato, on 
 the Properties of the Tetanus Poison The Investi- 
 gations of Bruschetini and of Camara Pestana on 
 its Diffusion in the Animal Body Vaccination 
 against Tetanus: Behring and Kitasato The New 
 Notions Contributed by these Authors. 
 
 We begin our exposition by the study of the 
 poisons produced by the bacillus of tetanus. These 
 poisons are not the first known, but they are in many 
 respects the best studied, of the microbian poisons. 
 
 Brieger was the first to investigate the toxic 
 products formed by the bacillus of tetanus. In the 
 cultures of this bacillus, as well as in the amputated 
 arm of a patient, he found several characteristic 
 ptomaines. There was tetanm, which has the prop- 
 erty of provoking in mice, even in very small doses, 
 
_ 8 9 - 
 
 attacks of trismus and of tetanus ending in death; 
 tetanotoxin, which provokes convulsive paroxyms, 
 followed by complete paralysis; spasmotoxin, which 
 causes cramps; and lastly, toxin, which has the prop- 
 erty of stimulating the salivary and lachrymal secre- 
 tion. 
 
 The researches of Brieger date from a period 
 when we did not know how to cultivate the tetanic 
 bacillus in a state of purity. Since then, Kitasato has 
 succeeded in making pure cultures of tetanus. One 
 may ask if in the pure cultures the ptomaines of 
 Brieger may be found ? Kitasato and Weyl have 
 put to themselves this question. They have suc- 
 ceeded, by employing Brieger's method, in finding in 
 notable quantity tetanin and traces of tetanotoxin. 
 Among the volatile products of tetanus cultures they 
 have found H 2 S, butyric acid, indol, and phenol. 
 But the two bases found were little toxic, and quite 
 insufficient to explain the disease. Tetanin does 
 not provoke convulsions and salivation in mice except 
 when given in very considerable doses, and spasmo- 
 toxin causes only paralysis.* 
 
 It was evident (and this is the conclusion of 
 these authors) that the true tetanus-poison must be 
 found elsewhere. 
 
 * Brieger: Ueber Ptomaine, iii, Theil. Berlin, 1886. 
 Kitasato and Weyl: Zur Kentniss der Anaeroben (Zeitschrift 
 f. Hygiene, t. viii, p. 404). 
 
90 
 
 Moreover, already in 1890 Faber had found in 
 tetanus cultures a poison of quite another nature than 
 the ptomaines of Brieger. In filtering cultures of the 
 tetanus bacillus through the Chamberland filter, 
 Faber obtained liquids deprived of bacilli, but repro- 
 ducing in animals all the symptoms of tetanus in the 
 same degree as the tetanus bacilli themselves. In- 
 toxication by the poison resembled infection by the 
 bacillus itself in the two following respects: (i) both 
 demand a certain time for their manifestation a 
 period of incubation for the intoxication as for the 
 infection; (2) when introduced into the stomach, the 
 chemical poison is quite as inoffensive as the living 
 bacillus. Faber also observed that the toxicity of the 
 filtered cultures was completely destroyed by a five- 
 minutes heating to 65 C., and even by the addition of 
 alcohol. 
 
 He concludes that the tetanus poison is not a 
 ptomaine, but rather a toxic diastase like that of 
 jequirity or that of diphtheria. Faber had not at his 
 disposal pure cultures of tetanus.* 
 
 But these results were soon confirmed by Tizzoni 
 and Cattani. These authors have found that the 
 tetanus bacillus produces its poison in pure cultures 
 on gelatin, and not in such as are made in bouillon. 
 The toxic substance of the filtered cultures is modi- 
 
 * Knud Faber: On the Pathogeny of Tetanus (Berlin. 
 Klin. Woch., 1890, No. 31). 
 
fied by precipitation by alcohol. It does not dialyze, 
 and this has enabled the authors to isolate it in the 
 following way: Filtered cultures are supersaturated 
 by ammonium sulphate in excess; a precipitate is 
 formed, which contains the toxic substance; this 
 precipitate is dissolved in water, freed from salts by 
 dialysis, and evaporated in a vacuum at a low tem- 
 perature. The toxic substance obtained in this way 
 presents itself as a yellow body, of crystalline aspect. 
 It is rendered completely inactive by thirty minutes' 
 heating up to 60 C. One hour's heating at 55 C. 
 reduces its toxicity. The alkalies, carbonic acid, 
 organic acids, and dilute mineral acids, do not modify 
 it. The concentrated mineral acids destroy its toxi- 
 city. 
 
 As to the nature of this tetanus poison, Tizzoni 
 and Cattani note numerous similarities with the 
 soluble ferments. In gelatin the tetanus bacillus 
 secretes a peptic ferment which it does not produce 
 in bouillon, and all the influences which modify the 
 tetanus poison destroy also the peptic diastase.* 
 
 In their researches on the bacterial poisons, 
 Brieger and Frankel speak of the toxalbumin of 
 tetanus. This toxic substance was soluble in water. 
 Its parent cultures were developed in sweetened 
 
 * Tizzoni and Cattani: Sul Veleno di Tetano (Riforma 
 Medica, 1890, No. 128). Tizzoni and Cattani: Experimentelle 
 Untersuchungen liber das Tetanusgift (Archiv f. Exper. 
 Pathol., Bd. xxvii). 
 
bouillon, then filtered through the Chamberland filter, 
 reduced by evaporation in a vacuum, and precipitated 
 by absolute alcohol.* Vaillard and Vincent have ob- 
 tained in tetanus cultures, made in bouillon and fil- 
 tered, an extremely active poison, of which -fa and y^ 
 c.c. sufficed to killed a guinea-pig. They have con- 
 firmed most of the foregoing data: thus, for instance, 
 the feeble resistance of the poison to heat, and its inac- 
 tivity in the digestive tube. Among the new data of 
 their work, I may cite especially the following facts: 
 Direct solar light destroys rapidly, in free air, the 
 toxicity of the filtered liquid. Acidification does 
 not modify its toxic power. Absolute alcohol does 
 not alter the toxic substance; it precipitates it in 
 part, and the product of this precipitation is tetani- 
 gen. The tetanus poison adheres to certain precipi- 
 tates which are produced in the liquids in which it is 
 contained: thus it is that the precipitates of phos- 
 phate of lime or of alum carry down a part of this 
 active substance, but this precipitation is quite in- 
 complete. Vaillard and Vincent believe that the 
 tetanus poison belongs to the diastases, f 
 
 Kitasato has made a detailed study of the tetanus 
 poison. He has determined the exact length of time 
 
 *Briegerand Frankel: Untersuchungen ttber Bacteri- 
 engifte (Bui. Kl. Woch., 1890, Nos. n and 12). 
 
 f Vaillard et Vincent: Le Poison Tetanique (C. R. de la 
 Societe de Biologic, 1890, Nov. 13). "Vaillard et Vincent: 
 Contribution a 1'Etude du Tetanos (Annales de 1'Institut 
 Pasteur, 1891, No. i). 
 
93 ~ 
 
 necessary for the different physical influences (light, 
 heat, etc.), or such chemical influences as acids and 
 alkalies to destroy the tetanus poison. He has found 
 that absolute alcohol added in sufficient quantity not 
 to leave the poison in solution destroys it. In general, 
 Kitasato has not found the means of isolating the 
 tetanus poison from the liquid in which it is con- 
 tained.* 
 
 Here end our sources of knowledge as to the 
 chemical nature of the tetanus poison. As to the 
 mode of its action on the animal organism, we have 
 not made much advancement since the researches 
 of Knud-Faber. We know that the poison repro- 
 duces exactly all the symptoms by tetanus in the 
 same way as we obtain them with the bacillus itself. 
 These phenomena of systemic poisoning do not ap- 
 pear immediately after the injection of the toxic 
 liquid, but after a certain time of incubation, which 
 diminishes as the dose or the toxicity of the in- 
 jected filtered liquid augments. 
 
 The morbid symptoms begin, in the case of sub- 
 cutaneous injection of the liquid, at or near the point 
 of inoculation. The first contractions appear in the 
 muscles nearest the seat of inoculation. Bruschetini 
 has studied the diffusion of the tetanus poison in the 
 animal organism; he finds that it takes place chiefly 
 
 * Kitasato: Experimentelle Untersuchungen tiber das- 
 Tetanusgift, t. x, p. 267. 
 
94 
 
 along the nervous system. He made use of filtered 
 cultures of tetanus.* 
 
 Camara Pestana, working with filtered cultures, 
 has carried out the following conclusions: 
 
 i st. The absorption of the toxine takes place by 
 the blood. 
 
 2d. The lungs, the spleen, the kidneys, but prin- 
 cipally the liver, take from the blood the toxic princi- 
 ple, and retain it. 
 
 3d. The toxine is not appreciably eliminated by 
 the urine. 
 
 4th. Despite the predominence of the neuro- 
 muscular phenomena, no one has yet succeeded in 
 detecting the presence of the toxine in the nervous 
 and muscular tissue, and all the experiments made 
 with these tissues have given negative results, f 
 
 The presence of the toxine in the blood of per- 
 sons affected with tetanus has also been noted by 
 Bruschetini, Kitasato, and Nissen. 
 
 The physiological study of the influence of the 
 tetanus poison on the different parts of the neuro- 
 muscular apparatus has not yet been made, and we do 
 not know how this poison acts. Faber has presented 
 
 * Bruschetini: Recherches Preliminaires sur la Diffu- 
 sion du Poison du Tetanus dans 1' Organisme (Annales de 
 Micrographie, 1890, Nov. 20). 
 
 f Camara Pestana: Diffusion du Poison du Tetanos dans 
 1'Organisme (C. R. de la Societe de Biologic, 1891, June 27). 
 
95 
 
 the hypothesis that it affects the terminal nerve-plates, 
 like curare. 
 
 With regard to the subject of immunity from 
 tetanus, there are some important details. At first, 
 all the experimenters arrived at constantly negative 
 results in trying to vaccinate animals against tetanus. 
 At length, Behring and Kitasato announced that they 
 had succeeded in rendering animals immune by vac- 
 cination. The animals vaccinated against tetanus ex- 
 emplified by their behavior the following facts, which 
 were quite new and important: 
 
 1. The hares which were vaccinated and resisted 
 the inoculation of the living tetanus bacillus, were 
 also refractory to the injection of the chemical poison 
 produced by this bacillus. 
 
 2. This insensibility of the vaccinated animals to 
 the poison is not due to an habituation of their organ- 
 ism to the poison, but to its destruction in their 
 bodies. In fact, the blood, and more particularly the 
 serum of the blood, of the vaccinated hares, when 
 mixed with the tetanus poison, even in small propor- 
 tion, destroys it. The blood of animals not vaccin- 
 ated does not possess this antitoxic property. 
 
 3. The serum of vaccinated animals possesses 
 not only in vitro the antitoxic property, it also exer- 
 cises it in the bodies of other animals. For the serum 
 of vaccinated hares, when injected in mice, renders 
 them refractory to subsequent inoculation of the bacil- 
 lus or of the tetanus-poison. It may even cure tetanus 
 in mice already sick. 
 
_ 9 6 - 
 
 We see that this work of Behring and Kitasato 
 contains three new and important notions: Vaccina- 
 tion against tetanus; the antitoxic action of the animal 
 humors; and immunization by these humors. These 
 three notions have been controlled and studied since 
 then by many experimenters, and in general have 
 been found perfectly exact. In the following chap- 
 ter we shall state the results acquired by these stud- 
 ies.* 
 
 * Behring et Kitasato: Ueber das Zustande-Kommen 
 der Diphteric immunitat und der Tetanus immunitat bei 
 Thieren (Deutsche Medic. Wochenschrift, 1890, No. 49). 
 
CHAPTER X. 
 
 THE POISONS OF TETANUS (Continued). 
 
 SUMMARY. Vaccination against Tetanus is Obtained 
 by Tizzoni, Cattani, and Vaillard The Researches 
 of Kitasato; of Behring; of Brieger, Kitasato, 
 and Wassermann Immunization The Labors of 
 Tizzoni, Cattani, and Vaillard The Researches of 
 Ehrlich; of Brieger and Ehrlich; of Brieger and 
 Frank Application of the Method of Behring and 
 Kitasato to the Treatment of Tetanus in Man. 
 
 We must remark that following the epoch of his 
 publication with Behring, Kitasato did not possess a 
 method of vaccinating animals against tetanus. He 
 had only at his command certain animals which were 
 shown to be fortuitously vaccinated; for a long time 
 he had been able to give only erroneous procedures 
 for this vaccination. 
 
 But independently of these researches other ex- 
 perimenters have succeeded in vaccinating animals 
 against tetanus. 
 
 Tizzoni and Cattani have vaccinated pigeons and 
 dogs against tetanus by injecting little doses of the 
 living cultures of tetanus. On these vaccinated ani- 
 mals they have been able to confirm the fundamental 
 
 7 OOO 
 
propositions of Behring and Kitasato respecting anti- 
 toxic reactions and immunization.* Vaillard, in util- 
 izing the method of C. Frankel (which we shall de- 
 scribe under the head of Diphtheria), has been able 
 to vaccinate hares. The method of Vaillard consists 
 in heating for an hour, at 60 C., filtered cultures of 
 tetanus. The tetanus poison is modified by heat to 
 the point of being unable longer to provoke tetanus 
 even when injected in large doses. But it acquires, by 
 heating, the property of conferring immunity. If we 
 heat the filtered cultures to 65 C., they no longer 
 confer immunity.! 
 
 KitasatoJ described later a process for vaccin- 
 ating against tetanus. It consists in inoculating hares 
 with the living and virulent cultures of tetanus, mixed 
 with progressively decreasing doses of bichloride of 
 iodine. But the results were first but little favorable, 
 for out of fifteen hares he succeeded in vaccinating 
 only six. Since then, however, the process with 
 ter-chloride of iodine (IC1 3 ) has been sensibly im- 
 proved at Koch's Institute by Behring, so far as to 
 
 *Tizzoni and Cattani: Ueber die Art einem Thiere die 
 Immunitat gegen Tetanus zu tibertragen (Centralblatt f. Bac- 
 teriologie, t. ix, No. 6). 
 
 f Vaillard: Immunite centre le Tetanos (C. R. de la 
 Soc. de Biol., 1891, Feb. 21). 
 
 \ Kitasato: Experimentelle Untersuchungen liber das 
 Tetanusgift (Zeitschrift f. Hygiene, t. x, p. 267). 
 
 Behring: Ueber Immunisirung und Heilung von Ver- 
 auchtieren beim Tetanus (Zeitschrift f. Hygiene, t. xii, p. 45). 
 
99 
 
 enable us to vaccinate without great danger all the 
 animals under experimentation: mice, hares, sheep, 
 and horses. Still this process makes the animals 
 quite sick for some time. Lastly, Brieger, Kitasato, 
 and Wassermann have found at Koch's Institute a 
 method of vaccination perfectly inoffensive. Resum- 
 ing the already antiquated researches of Wooldridge 
 on cultures of the microbes in extract of thymus, and 
 following exactly his indications, these authors have 
 made, with the thymus,* culture-media which have 
 given them interesting results with the different 
 microbes, especially with the bacillus of tetanus. 
 These tetanus cultures made in extract of thymus are 
 asporogenous and very slightly toxic. On the other 
 hand, the extract of thymus mixed with a filtered cul- 
 ture of tetanus made in ordinary bouillon, and very 
 toxic, destroys little by little the toxicity of the 
 culture product. In employing these mixtures when 
 two days old in progressively increasing doses, the 
 authors have succeeded in vaccinating without dan- 
 ger and without apparent trouble, animals, such as 
 mice, the most sensitive to tetanus.f 
 
 Vaillard has recently come back to the question 
 
 * Wooldridge: Versuche iiber Schuzimpfung auf Chem- 
 ischen Wege (Archiv f. Anatomic und Physiologic, Physio- 
 logische Abtheilung, 1888, p. 527). 
 
 f Brieger, Kitasato, and Wassermann: Ueber Immuni- 
 tat und Giftfestigung (Zeitschrift f. Hygiene, t. xii, p. 
 1887). 
 
100 
 
 of vaccination against tetanus.* Besides his old 
 method of vaccination by cultures filtered and heated 
 to 60 C., he indicates two new processes: one of these 
 is very nearly like that of Behring and Kitasato, and 
 consists in injecting in animals tetanus-cultures to 
 which iodine solution has been added; the other is- 
 identical with that of Tizzoni and Cattani inocula- 
 tion with extremely small doses of the virus. Behring 
 has also utilized with success this process of vaccina- 
 tion after dilution. 
 
 It will be seen that this question of vaccination 
 has gone through three successive stages. At first, 
 experimenters only succeeded in vaccinating a certain 
 proportion of the animals under trial, while the others 
 died in consequence of the manipulations. Then they 
 obtained more constant results, but the vaccinated 
 animals suffered more or less severely in their general 
 health (local tetanus, fever, and emaciation). At last 
 they found a method both sure and inoffensive (Brie- 
 ger, Kitasato, and Wassermann). The antitoxic prop- 
 erties of the serum of vaccinated animals were soon 
 found by all the experimenters. 
 
 Tizzoni and Cattani have made a special study 
 of the antitoxic substance of serum. They have 
 found that their antitoxine is enfeebled by heating to 
 65 C. half an hour, and completely destroyed by 
 heating it the same length of time at 68 C. (the 
 
 * Vaillard, Annales de 1'Institut Pasteur, 1892, No. 4. 
 
101 
 
 temperature of coagulation of the serum). They 
 have found also that this antitoxic substance does 
 not dialyze; that it is destroyed very quickly by hy- 
 drochloric acid, by lactic acid in great excess, and 
 by the alkalies. It is precipitated by ammonium sul- 
 phate in supersaturation, and also by absolute alco- 
 hol. From this latter precipitate it can be extracted 
 by water or by glycerin.* 
 
 Vaillard has studied especially the relations be- 
 tween the immunity of animals and the antitoxic 
 property of this serum. He has been led by his re- 
 searches to deny the possibility of explaining immu- 
 nity by the antitoxic property of serum. In animals 
 naturally refractory, as, for instance, hens, the serum 
 in the natural state is not antitoxic; it only acquires 
 this property after the injection of a large dose 
 of tetanus poison. Likewise, in the vaccinated ani- 
 mals, one may not find the antitoxic state of the 
 serum. This state only appears as the result of the 
 action of large doses of the soluble products of the 
 tetanus bacillus. The antitoxic action of the serum is 
 only a contingent property of the refractory animals. 
 It cannot, therefore, serve to explain natural or ac- 
 quired immunity. We must add that Vaillard did 
 not succeed in curing tetanus by the serum of his re- 
 fractory animals. He found also that the spleen and 
 
 *Tizzoni et Cattani: Ueber die Eigenschaften des Te- 
 tanus Antitoxins (Centr. f. Bacteriologie, t. ix, No. 21). 
 
IO2 
 
 aqueous humor of the vaccinated animals did not 
 possess antitoxic properties. Their muscles, however, 
 possess these properties, at least in vitro* 
 
 Apropos of the spleen, we must speak of the re- 
 searches of Tizzoni and Cattani, who have found that 
 it is impossible to vaccinate against tetanus animals 
 that are deprived of their spleen. 
 
 The third thesis of Behring and Kitasato vac- 
 cination and cure by the serum of vaccinated animals 
 has also been contested. While succeeding in pre- 
 venting the disease by the serum, neither Tizzoni and 
 Cattani, nor Vaillard, succeeded in curing it. 
 
 Many other contradictions have been affirmed 
 against the general theory of immunization contra- 
 dictions due for the most part to the partisan influ- 
 ence of the old theories of immunization; but all 
 these contradictions have received their quietus, 
 thanks especially to the interesting researches of 
 Ehrlich.f Experimenting with abrine and ricine, two 
 toxic albuminoids of vegetable origin, Ehrlich dem- 
 onstrated numerically that the immunity of animals 
 against the poison is not an invariable quantity, but 
 may have very different degrees. He demonstrated, 
 moreover, that with the degree of the immunity 
 
 * Vaillard: Property of the Serum of Animals Refrac- 
 tory to Tetanus (C. R. de la Societe de Biologic, June 6, 
 1891). 
 
 f Ehrlich, Deutsche Med. Wochenschrift, 1891, Nos. 32 
 and 44. 
 
103 ~ 
 
 acquired by the vaccinated animals varies also the 
 antitoxic and immunizing power of their serum. 
 
 Thus were cleared up all the previous difficulties; 
 the experimenters who could not cure, but who vac- 
 cinated only with the serum of vaccinated animals, had 
 evidently not obtained in those animals the grade of 
 immunity sufficient for their serum to be curative. In 
 fact, in continuing their experiments, Tizzoni and 
 Cattani, and Vaillard, have perfectly succeeded in 
 curing their tetanic animals with the serum of the 
 vaccinated animals. At the present time, there re- 
 mains no longer any doubt as to the reality of the 
 third thesis of Behring and Kitasato; we can pre- 
 vent and we may cure tetanus by the serum of ani- 
 mals vaccinated against this disease. 
 
 Erhlich has contributed other interesting docu- 
 ments to this question of the antitoxines. He has seen 
 that if we vaccinate against one of the vegetal toxines 
 pregnant mice, and give them after they have dropped 
 their young the young of non-vaccinated mice to 
 suckle, these little mice become in their turn refrac- 
 tory to the given poison.* 
 
 These facts are of the highest importance, for 
 they demonstrate that the immunizing substance is 
 secreted in the milk, and that it is absorbed by the 
 digestive canal. Ehrlich has applied this datum to 
 the infectious diseases. 
 
 Ehrlich, Zeitschrift f. Hygiene, t. xii, p. 183. 
 
104 
 
 He has been able to prove that the milk secretes 
 antitetanine, and in that condition, being ingested, 
 confers immunity against tetanus. With Brieger he 
 has made other experiments on a large scale. A 
 pregnant goat was by them vaccinated against teta- 
 nus, according to the method of Brieger, Kitasato, 
 and Wasserman. Later on, the goat's milk, injected 
 in mice, immunized them perfectly against tetanus. 
 
 Behring and Frank have recently published some 
 interesting researches on the properties of the sub- 
 stance which immunizes against tetanus.* They 
 found that the serum of a vaccinated horse kept for 
 two months its immunizing power, although preserved 
 (with a >^-per-cent. solution of phenic acid) in a flask 
 closed with a glass stopper, with no other precaution 
 against the entrance of the air or of microbes. They 
 found also that this immunizing power of serum is 
 not abolished by dilution with distilled water, or by 
 heating it twenty-five minutes at 65 C. when it 
 begins to coagulate. They insist also on the fact 
 that to cure tetanus when declared, it is necessary to 
 employ stronger doses of the serum than would be 
 required for propyhlaxis. 
 
 We must say a few words, in conclusion, about 
 the therapeutic attempts made on man with the im- 
 munizing serum. Kitasato was the first to attempt 
 
 * Behring and Frank, Deutsche Medic. Wochenschrift, 
 1892, No. 21. 
 
105 
 
 to cure tetanus in man by the serum of the vaccinated 
 hare. He employed very feeble doses, and his at- 
 tempt was not crowned with success. Tizzoni and 
 Cattani, on the contrary, report already seven cases 
 of cure by their antitetanine, prepared with the serum 
 of vaccinated dogs. 
 
 More recently, this treatment was attempted 
 without success in two cases of human tetanus in 
 Paris.* It is evident that there have as yet been too 
 few cases to warrant speaking for or against the 
 method of Behring and Kitasato. But the brilliant 
 results of experimentation leave no doubt that success 
 on man is only a question of method and of doses. 
 
 *Annales de 1'Institut Pasteur, 1892, No. 4. 
 
CHAPTER XL 
 
 THE POISONS OF DIPHTHERIA. 
 
 SUMMARY. The Researches of Roux and Yersin; of 
 Loffler C. Frdnkel Succeeds in Vaccinating Animals 
 against Diphtheria The Researches of Behring 
 and His Discovery of Immunization The Diffi- 
 culties Not Yet Surmounted of Diphtheritic Vac- 
 cination. 
 
 In filtering the diphtheritic cultures through the 
 Chamberland filter, Roux and Yersin obtained toxic 
 liquids. These filtered cultures injected in animals 
 produce the same series of symptoms as the living 
 diphtheria bacillus. In guinea-pigs these filtered cul- 
 tures generally cause the following lesions: At the 
 place of inoculation is found an exudative oedema; 
 the lymphatic glands are congested; the small intes- 
 tine, the lungs, and the suprarenal capsules are en- 
 gorged; and the pleura contains a serous effusion. 
 The systemic intoxication of hares is characterized 
 especially by diarrhoea and fatty degeneration of the 
 liver. In smaller doses which kill only after a long 
 time, the poison of the filtered cultures may provoke 
 in animals the different paralyses which are so fre- 
 quent in diphtheria in man. 
 
107 
 
 The toxic substance is completely destroyed by 
 heating for ten minutes up to 100 C.* 
 
 In another memoir, Roux and Yersin have 
 studied at greater length the properties of this diph- 
 theritic poison, which they have classed among the 
 diastases f We shall not enter farther into the discus- 
 sion as to the chemical nature of this poison a discus- 
 sion which is set forth at length with all its biblio- 
 graphical details in Chapter VI. 
 
 Loftier has confirmed the existence of the diph- 
 theritic poison. He extracted it by means of glycerin 
 from cultures made on hashed meatj 
 
 The resistance of the diphtheria poison to the dif- 
 ferent means of destruction has been but little studied. 
 Heat above 60 C. destroys it, or, as we believe, mod- 
 ifies it, leaving the cachectizing poison uninjured. 
 The same decomposition is obtained by the action of 
 the peptic diastases, such as trypsin and pepsin. Con- 
 tact with alcohol modifies also the diphtheritic poison. 
 The different oxidizing agents, as potassium perman- 
 ganate, destroy it completely. The reducing agents, 
 such as HS 2 , have no action on it. 
 
 Roux and Yersin have completely failed in their 
 
 *Roux and Yersin, Annales de 1'Inst. Pasteur, 1888, 
 No. 12. 
 
 f Roux and Yersin: Study of Diphtheria (Annales de 
 1'Inst. Pasteur, 1889, p. 273). 
 
 | Loffler, Deutsche Medic. Wochenschrift, 1889, Nos. 
 5 and 6. 
 
io8 
 
 attempts to habituate or to vaccinate animals against 
 the diphtheritic poison. C. Frankel has been more 
 successful. In heating cultures of diphtheria at 70 
 C, he has succeeded in depriving them in great part 
 of their toxicity, while not injuring their vaccinal 
 power. He has found that the heating of diphtheritic 
 cultures three weeks old should be made at the tem- 
 perature of from 60 to 70 C. If this temperature is 
 exceeded, you obtain no longer vaccinant effects, and 
 you even produce a slow intoxication of the animals. 
 With cultures heated to 70 C., and inoculated in the 
 dose of from 10 to 20 cubic centimeters under the 
 skin of guinea-pigs, immunity is conferred at the end 
 of two weeks.* 
 
 Shortly after Frankel's communication, appeared 
 the memoir of Behring on the same subject. Behring 
 gives five different processes of vaccinating animals 
 against diphtheria: 
 
 1. With the sterilized cultures, as in the method 
 of Frankel. 
 
 2. With diphtheritic cultures to which has been 
 added trichloride of iodine. 
 
 3. By the pleuritic exudation of guinea-pigs which 
 have died of diphtheria. 
 
 4. By the inoculation of animals with the virulent 
 diphtheritic bacillus followed by their treatment by 
 trichloride of iodine in subcutaneous injection. 
 
 * Vide Gamalela, in C. R. de la Soc. de Biologic, Feb. 
 20, 1892; also C. Frankel, Berlin. Klin. Woch., 1890, No 49. 
 
109 
 
 5. By oxygenated water, which has the property 
 of vaccinating the animals against diphtheria, just as 
 trichloride of iodine vaccinates them against tetanus. 
 
 The great number of processes which were only 
 touched upon in the memoir of Behring made it plain 
 that this author did not possess a good method of vac- 
 cination against diphtheria.* And his subsequent pub- 
 lications only went to confirm this conviction. 
 
 Proskauer and Wassermann have attempted to 
 vaccinate against diphtheria by the poisons subjected 
 to heat, but with constantly negative results. f 
 
 Zimmer, at the laboratory of C. Frankel, has veri- 
 fied the exactitude of the assertions of Behring. He 
 has noted, in general, that a more or less complete 
 vaccination may result from the employment of these 
 different processes, but that no one of them is certain. 
 With most of the processes of Behring the immunity 
 of the animals is the exception, and an ultimate death 
 is the rule. 
 
 Only one of the methods of Behring should be 
 signally excepted: it is that of the injection in the 
 animals of diphtheritic cultures attenuated by the 
 addition of trichloride of iodine. This process has 
 given Zimmer positive results. J It is far, however, 
 
 * Behring, Deutsche Medic. Woch., 1890, No. 50. 
 f Proskauer and Wassermann, Deutsche Med. Woch.. 
 1891, No. 17. 
 
 \ Zimmer, Deutsche Medic. Woch., 1892, No. 16. 
 
no 
 
 from constituting a definitive method of vaccination, 
 for Behring himself does not trust it; in his most re- 
 cent work on this subject he describes as his method 
 of anti-diphteritic vaccination certain extremely com- 
 plicated processes. He injects successively diphthe- 
 ritic cultures heated to 90, 80, and 70 C.; then 
 mixtures, at different degrees of strength, of cultures 
 with trichloride of iodine. Despite this long and 
 complicated preparation, you will note in his experi- 
 ments that most of his vaccinated animals die of 
 diphtheria. Moreover, Behring himself acknowledges 
 that he does not possess a satisfactory method of 
 vaccination. 
 
 We shall see further on, what enormous interest 
 is attached to this problem of how to find a sure 
 method. The principles which guide Behring in his 
 vaccination procedures are the following: 
 
 To give immunity to receptive animals, and to 
 augment immunity in animals which already pos- 
 sess it in a certain degree, the vaccinal injections 
 ought to be followed by a reaction of the organism. 
 This reaction is general and local.* Behring does 
 not indicate in what the first consists; as to the local 
 reaction, it is a tumor more or less circumscribed. If 
 the vaccinal reaction is nil, the vaccinal injection will 
 
 * Behring and Wernicke, Zeitschrift fiir Hygiene, t. 
 xii, p. 10. Also, Gamaleta's articles in Ann. de 1'Inst. Pas- 
 teur, 1888, No. 10, and Centralblatt f. Bacteriologie, 1888, t. 
 iv, p. 161. 
 
Ill 
 
 not give any immunity, or will not augment the pre- 
 existing immunity. If the reaction is excessive, the 
 vaccinal injection leads to an opposite effect; it aug- 
 ments the receptivity of the animal to diphtheria. If 
 the reaction is moderate, the animals acquire a cer- 
 tain degree of immunity, but only at the end of a 
 certain time several weeks or several months. Ac- 
 cording to this exposition, you can see with what 
 difficulty and slowness vaccination is made by the 
 procedures of Behring. 
 
 Consequently, it is with reason that the authors 
 of a recent work on the subject Brieger, Kitasato, 
 and Wassermann declare that there does not thus far 
 exist any process which enables us to vaccinate with 
 certainty against diphtheria. In making use of Wool- 
 dridge's method, they give, in their turn, a new pro- 
 cedure: they cultivate the diphtheritic bacillus in 
 extract of thymus, and then heat this culture (after 
 C. Frankel) between 60 and 70 C. for fifteen min- 
 utes. With the vaccine prepared in this way, they 
 have obtained some positive results. But this mixed 
 procedure is not more sure.* 
 
 Although we do not yet possess a satisfactory 
 method of vaccination against diphtheria, the fact of 
 the possibility of this vaccination is sufficiently estab- 
 lished. 
 
 *Breiger, Kitasato, and Wassermann, Zeitschrift fttr 
 Hygiene, t. xii, p. 137. Also, Behring and Kitasato, Deutsche 
 Medic. Wochen., 1890, No. 49. 
 
112 
 
 The study of these vaccinated animals has led 
 Behring and Kitasato to results extremely interesting. 
 
 Behring* has noted and this fact has been con- 
 firmed by other observers that the inoculation of 
 the diphtheritic bacillus in vaccinated animals is 
 followed by the formation of a necrotic patch under 
 which the bacillus long lives in a state of activity. It 
 is not, we perceive, destroyed in the vaccinated organ- 
 ism. It does not even seem to have lost its virulence, 
 for its subsequent inoculation in animals not vaccin- 
 ated provokes in them typical diphtheria. As, how- 
 ever, it remains inoffensive to the vaccinated organism 
 which gives it harbor, Behring supposed that the lat- 
 ter possesses the power of destroying its poison. 
 
 We have seen in the history of the tetanus poison 
 how this supposition was confirmed by Behring and 
 Kitasato. 
 
 For diphtheria, as for tetanus, these authors found 
 in the serum of vaccinated animals certain remarkable 
 antitoxic properties. The serum of certain animals 
 vaccinated against diphtheria guinea-pigs, hares, 
 sheep has the power to destroy in vitro the diph- 
 theritic poison; it may consequently prevent and cure 
 diphtheria in animals in which it is injected in suffi- 
 cient quantity. 
 
 If all these researches have not yet led to any 
 
 * Behring: Communication to the Congress of Hygiene. 
 London, 1891. 
 
good practical result in the cure of diphtheria in 
 man, this is due solely to the difficulty of procuring 
 the serum of vaccinated animals in great quantity; 
 and this difficulty depends on the absence of a good 
 method of vaccination. It is for this reason that we 
 have here dwelt at such length on the question of 
 
 vaccination against diphtheria. 
 8000 
 
CHAPTER XII. 
 
 THE POISONS OF CHOLERA AND OF THE 
 AVICIDE VIBRIO. 
 
 SUMMARY. The Researches of the Author on the Poi- 
 sons of Cholera and of the Avicide Vibrio The 
 Researches of Hernandez and Bruhl, of Niessen 
 and Behring, and of Zasslein. 
 
 The history of the development of our sources of 
 information respecting the poisons of cholera, shows 
 in a typical manner the struggle of the different con- 
 ceptions concerning the chemical nature of the micro- 
 bian poisons. In order to explain cholera, we have 
 seen successively advanced such considerations as 
 the products of the metabolism of the Indian vibrio, 
 the diastases which it secretes, and, lastly, the action 
 of the microbe by its own substance.* Cantani was 
 the first to indicate that the cholera poison may be 
 neither a diastase nor a ptomaine, but the cholera 
 vibrio itself. This he has not proved, f 
 
 I have myself found the two cholera poisons: the 
 primary poison and the modified poison. The first, 
 
 * Gamalela: Experimental Researches on the Poisons of 
 Cholera (Arch, de Med. Exper., March i, 1892). 
 
 t Cantani, Deutsche Medic. Wochen., 1886, No. 45. 
 
which we believe to be a nucleo-albumin, has the 
 property of provoking a systemic intoxication ex- 
 ceedingly like the symptoms of Asiatic cholera. In 
 hares this intoxication manifests itself chiefly by a 
 prolonged and violent diarrhoea; in dogs the vomit- 
 ings predominate, and may last several hours. In all 
 cases the poison determines a violent lesion of the 
 gastro-intestinal tube. 
 
 This poison is extremely fragile. It is destroyed 
 by heat above 60 C., by alcohol, and by the strong 
 alkalies. It is carried down by the different precipi- 
 tates which may be produced in the cholera cultures; 
 for example, by that which is formed by acetate of 
 lead. It may be extracted from them by alkalinized 
 water. 
 
 What is the mode of action of this poison ? By 
 some of its reactions it resembles the soluble fer- 
 ments. We know also that the cholera vibrio may 
 produce the different diastases. It may be asked if 
 it is not these diastases which are toxic in the cultures 
 of cholera? Some think that the excessive produc- 
 tion of liquid in cholera stools is the result of a hyper- 
 secretion of the intestinal glands (Cohnheim), and we 
 know that the substances secreted by the glands 
 stimulate their activity if injected into the blood. In 
 studying the toxic action of the cultures of different 
 microbe producers of diastases, we find, in fact, that 
 they all have as a common character a certain 
 diarrhceic action, though this is most marked in 
 
n6 
 
 the cholera cultures. Even certain samples of pan- 
 creatin, in our experiments produced diarrhoea in 
 hares. But the results with this latter substance have 
 not been constant, and we have at least succeeded in 
 procuring some trypsin which is extremely active as a 
 peptic diastase and which has no diarrhceic action. 
 Many other ferments which we have studied in this- 
 connection were also devoid of the property of pro- 
 voking diarrhoea. 
 
 We must conclude that the diarrhceic poison of 
 cholera does not belong to the category of these fer- 
 ments. 
 
 The diarrhceic action is very general in the class- 
 of microbian poisons, but it is due to another mechan- 
 ism than that indicated above. 
 
 We have found still another cholera poison, 
 which is very stable. It belongs to the class of our 
 modified poisons, or nucleines, which Buchner has de- 
 nominated proteins or alkali-albumins. This poison 
 has no other interest than that of being associated 
 with the vaccinal substance of cholera. But we shall 
 not here set forth the details of cholera vaccination,, 
 which is still under controversy. There exists a 
 microbe which resembles in all respects that of 
 cholera. It is the avicide vibrio, which I discovered 
 at Odessa. Chemical vaccination with this microbe 
 is at present the best known of all the vaccinations. 
 As, besides, it has many analogies with cholera vac- 
 cinations, we shall venture to say a few words about it. 
 
The vaccinal substance or substances which are 
 potent against the avicide vibrio are also associated 
 with the nucleine or modified poison of the vibrio. 
 They may be isolated in two ways. First, if you sub- 
 ject to distillation in a vacuum the vibrionic cultures, 
 the substances which pass over have a manifest vac- 
 cinal action, while the toxic nucleine remains in the 
 residue. Now Briihl has found that if we precipitate 
 the toxic substance by acetate of lead, the filtrate 
 vaccinates perfectly. The vaccinal action of the 
 volatile products would lead one to think that the 
 vaccine is a ptomaine, but the ordinary methods for 
 the isolation of the ptomaines have resulted only in 
 inactive substances (Bruhl). Alcohol especially seems 
 to enfeeble the vaccinal properties of the vibrionic 
 cultures. 
 
 The study of the serum of animals vaccinated 
 against either the avicide vibrio or the cholera vibrio 
 reveals certain remarkable peculiarities. While the 
 serum of ordinary guinea-pigs favors the growth 
 of the avicide and cholera vibrios, the serum of 
 guinea-pigs vaccinated against these diseases acquires 
 the property of completely destroying these microbes. 
 We have no need of insisting upon the bearing of 
 these facts on the true solution of the problem of im- 
 munity. 
 
 Still another fact of considerable theoretical im- 
 portance has been found in connection with the dis- 
 eases caused by these two vibrios. Guinea-pigs which 
 
n8 
 
 are vaccinated against the avicide or cholera vibrio 
 are not more resistant to the vaccinal poison than 
 non-vaccinated guinea-pigs. This fact is of funda- 
 mental interest in reference to the entire mechanism 
 of vaccination. 
 
 Denied at first by certain bacteriologists, it was 
 later affirmed by the same, and adduced in opposition 
 to the great principles established for tetanus and 
 diphtheria. We have seen that in these two diseases 
 the animals which are vaccinated against the living 
 microbe become also refractory to the microbian 
 poisons. 
 
 Cholera and the vibrionic septicaemia seemed to- 
 be an exception to this rule, for in these two diseases 
 the animals succumb to the same dose of the poison 
 as the control animals. But this exception is only 
 apparent, and it disappears when we recognize the 
 distinction between the primitive and the modified 
 poisons of bacteria. 
 
 Towards the primitive poisons of cholera and of 
 septicaemia, the vaccinated animals are quite as re- 
 fractory as is the case with diphtheria and tetanus. 
 They are not refractory towards the modified poisons; 
 but as these latter are artificial products, this has 
 nothing to do with the theories of immunity. 
 
 With regard to the mechanism of vaccination, it 
 may appear strange that the modified poisons to 
 which the animal is not habituated render it refrac- 
 tory to the primitive poisons and to the bacteria. 
 
n 9 
 
 Here also we must make a distinction. The vaccinal 
 substance is not the modified poison, for it may be 
 separated from it. Not being toxic, it does not vac- 
 cinate the animals by the fact of habituation, but 
 probably by forming in their bodies a compound 
 which has antitoxic properties. 
 
r 
 
 CHAPTER XIII. 
 
 THE POISONS OF TUBERCULOSIS. 
 
 SUMMARY. The Researches of Koch, of Maffuci, of 
 Prudden and Ifodenpyl, of Straus and the Author, 
 of Grancher and Ledoux-Lebard The Tubercu- 
 lin of Koch, and the Researches which it has Pro- 
 voked The Toxomucin of WeyL 
 
 The tuberculous poisons have only lately been 
 found. Bacteriologists were long and unsuccessfully 
 searching for the toxic substances in the cultures 
 of Koch's bacillus, and they have only found them by 
 applying directly to the bacillus itself. 
 
 Koch has seen that the bacillus tuberculosis 
 when dead provokes by subcutaneous injection the 
 formation of abscesses in animals. Moreover, he has 
 obtained from the bodies of the bacilli a substance 
 called tuberculin, which has long been believed to 
 exercise a specific action on the tuberculous.* 
 
 Maffuci has found a cachectizing influence in 
 tuberculous cultures dead by age or killed by dis- 
 continuous sterilization.f 
 
 * Koch, Deutsche Medic. Wochenschrift, Jan. 18, 1891. 
 f Maffuci, Centralblatt f. Allgemeine Pathol., Dec. 15, 
 1890. 
 
121 
 
 Prudden and Hodenpyl have seen that the bacilli 
 when boiled and well washed may give rise to the 
 formation of curable nodular lesions.* 
 
 Straus and I have shown that the cadavers of 
 tubercle bacilli provoke in different animals a disease 
 which in its course and its lesions is extremely like 
 tuberbulosis produced by living bacilli. This disease, 
 which Grancher and Ledoux-Lebard have propossd 
 to call necro-tuberculosis,f is characterized by the 
 development in inoculated animals of a great sensi- 
 tiveness towards a new inoculation of tuberculosis. 
 This predisposition manifests itself by rapid death 
 after re-inoculation. The inoculation of dead bacilli 
 produces such changes in the animal organism that 
 the animals die very speedily after a new injection of 
 tuberculosis, an injection which produces only tardy 
 effects in healthy animals. 
 
 Another symptom of necrotuberculosis, but much 
 more apparent, is the continuous and progressive 
 emaciation of the animals. They may lose as much 
 as one-half of their initial weight. They die in a pro- 
 found cachexia. At the autopsy we find characteristic 
 lesions. 
 
 The different organs, and especially the lungs, in 
 
 * Prudden and Hodenpyl, New York Medical Journal, 
 June 6 and June 20, 1891. 
 
 . f Straus and Gamalela, Archives de Medecine Experi- 
 tnentale, 1891, No. 6. Grancher and Ledoux-Lebard, Arch, 
 de M6d. Exper., 1892, No. I. 
 
122 
 
 the case of intravenous injection of dead bacilli, are 
 strewn with granulations formed of epithelioid and 
 embryonal cells, and containing tubercle bacilli. The 
 volume of the nodules which may be thus produced 
 in the different organs depends almost exclusively on 
 the size of the mass of cadavers which has been inoc- 
 ulated. It is very interesting to note that the animal 
 organism cannot dislocate the bacterial colonies which 
 are inoculated into it. 
 
 When once the existence of the tuberculous poi- 
 son, and its presence in the cadavers of the bacilli, 
 was established, it was necessary to seek to determine 
 the toxic substance and to isolate it from the bacilli. 
 
 There were evidently different means for pro- 
 ceeding to this extraction. By alcohol and ether 
 Hammerschlag has extracted from the bacilli, in addi- 
 tion to lecithin and fat, a toxic substance which kills- 
 animals with convulsions.* Later, Zuelzer also suc- 
 ceeded in finding a convulsivant ptomaine in tubercu- 
 lous cultures.f 
 
 Much more interesting was Koch's discovery. 
 By means of glycerin-water at a boiling temperature 
 and in a neutral medium, Koch has extracted from 
 tubercle bacilli an albuminoid substance called tuber- 
 culin, which provokes in tuberculous animals fever 
 
 * Hammerschlag, Correspondenzblatt fur Schweizer- 
 Srzte, Oct. 15, 1888, and Centralblatt f. Klin. Med., Jan. I, 
 1891- 
 
 f Zuelzer, Berlin. Klin. Wochen., Jan. 26, 1891. 
 
123 
 
 and inflammation of the bacillary foci. Koch has 
 studied in detail the many very interesting properties 
 of this tuberculin.* Tuberculin dissolved in glycerin- 
 water is a very stable body which supports a heat of 
 1 60 C. It is easily precipitable by alcohol. 
 
 But if we attempt to purify it and to free it from 
 glycerin and salts, its properties are modified. Aque- 
 ous solutions of purified tuberculin are very unstable 
 in absence of glycerin. They easily give a precipitate 
 more or less abundant of a substance, insoluble in 
 water but soluble in alkalies, which preserves the toxic 
 action of the tuberculin. In absence of salts, the 
 purified tuberculin is no longer precipitated by abso- 
 lute alcohol. 
 
 The purified tuberculin has all the reactions of 
 albuminoid substances. It gives a precipitate by 
 acetic acid which is soluble in excess of the reagent. 
 It contains a great quantity of phosphorus. But it is 
 evidently not a pure substance, for it leaves from 14 
 to 21 per cent, of ashes, and then its mode of prepa- 
 ration by the concentration of cultures made in 
 bouillon adds to the extract of bacillary bodies all 
 the substances pre-existing in the bouillon. 
 
 Hunter f has endeavored to advance still farther 
 our knowledge of tuberculin. Unfortunately he also 
 
 * Koch, Deutsche Medic, Wochen., 1891, No. 43. Gama- 
 lela, Arch, de Med. Exper., 1891, No. 2. 
 
 f British Medical Journal, July 28, 1891. 
 
124 ~ 
 
 made use, in his chemical researches, of the crude 
 tuberculin of Koch, prepared by means of culture 
 bouillons, instead of taking the bacilli themselves, 
 washed and cleaned. In this crude tuberculin Hunter 
 found the four albumoses of Kuehne and Chittenden. 
 It is to be regretted that he did not attempt to find 
 them in the primitive bouillon of the cultures. 
 
 Hunter has also compared the differences in the 
 action on the tuberculous man and animal of the dif- 
 ferent modified tuberculins. The modification A, 
 which is the precipitate obtained by absolute alcohol, 
 has the property of producing the local reaction with 
 very little fever. The modification C, which contains 
 the substances not precipitated by absolute alcohol, 
 produces chiefly a high fever. The modification B, 
 which is the precipitate obtained by supersaturation 
 of the tuberculin with ammonium sulphate, possesses 
 the power of provoking the local and salutary reac- 
 tion without constitutional disorders. Lastly, the 
 modification CB has all the curative virtues of Koch's 
 tuberculin, without possessing the disadvantages 
 either the local inflammatory or the general febrile 
 reaction. This happy modification, CB, is obtained 
 by getting rid, by absolute alcohol, of most of the 
 albumins, and by the dialysis of other injurious sub- 
 stances. These results of Hunter are founded on an 
 altogether insufficient number of experiments. Fur- 
 thermore, according to all the other experimenters, 
 and particularly Pfuhl, the collaborator of Koch, tuber- 
 
125 
 
 culin has not any curative properties, especially if you 
 prevent the local reactions.* 
 
 The tuberculocidin of Klebs, prepared by means 
 of tuberculin, is entitled to no greater confidence. 
 
 As to the chemical nature of tuberculin, we can- 
 not pronounce with certainty until we have a purer 
 preparation. There is no doubt that it belongs to 
 our class of modified poisons, which we suppose to be 
 nucleines or nucleinic acids. It is evident, also, that 
 in Koch's tuberculin is not found the primary tuber- 
 culous poison. The action of tuberculin does not 
 explain the effects produced by the dead bacilli. 
 This action is not characteristic of tuberculin, for it 
 is also provoked by many other bacterian extracts. 
 The effects proper to the tuberculous poison con- 
 tained in the microbian cadavers, the predisposition 
 which it creates, and the suppurative and caseous 
 lesions, cannot be produced by tuberculin. 
 
 More important in this respect is the substance 
 which Weyl has extracted from tubercle bacilli by 
 means of a solution of caustic soda. This substance, 
 insoluble in acetic acid, is classed by Weyl among the 
 mucins, but it does not form a reducing substance, 
 and it contains phosphorus. It may be nucleo-albu- 
 min in the sense of Kossel and Hammersten. 
 
 This mucin of Weyl has a toxic action. Bjr 
 subcutaneous injection it produces in mice and 
 
 * Pfuhl, Zeitschrift ftir Hygiene, t. xi, page 241. 
 
126 
 
 guinea-pigs a necrosis of the skin. But the toxi- 
 cological study of Weyl is too incomplete. We cannot 
 but suppose that he had under observation the actual 
 primary tuberculous poison, that which produces the 
 local lesions of tuberculosis.* 
 
 *Weyl, Deutsche Medic. Wochen., Jan. 12, 1891. 
 
CHAPTER XIV. 
 
 THE POISONS OF CHARBON AND OFGLANDERS. 
 
 SUMMARY. Insufficiency of our Toxicological Knowl- 
 edge respecting Charbon and Glanders Different 
 Writings on the Subject. 
 
 Charbon (splenic fever, wool-sorters' disease, 
 deadly anthrax), which once had so great a doctrinal 
 importance as to guide the first steps of students in 
 bacteriology, has not preserved this eminent place 
 in toxicological researches. 
 
 Toussaint believed that he could confer immu- 
 nity on sheep and young dogs by means of anthracoid 
 blood freed from living bacteria by filtration or by 
 heating to 55 C., and even by antiseptics. 
 
 Chauveau brought powerful arguments in favor 
 of this chemical vaccination.* 
 
 After having, with Pasteur, denied its reality, 
 Chamberland and Roux published later some experi- 
 ments which showed that it is in fact possible to vac- 
 cinate sheep against charbon by means of soluble 
 substances, f 
 
 *Annales de 1'Institut Pasteur, 1888, No. 2. 
 \ Chamberland and .Roux, Ann. de 1'Institut Pasteur, 
 1888, No. 7. 
 
128 
 
 Already, moreover, before these experiments, this 
 possibility had been demonstrated by Wooldridge. 
 He found that anthracoid cultures made in a solution 
 of the substance which he called tissue-fibrinogen* 
 might confer immunity on hares if injected subcuta- 
 neously, when freed from living microbes by ebullition 
 or by filtration. What is surprising in the experi- 
 ments of Wooldridge is that his substance conferred 
 immunity not only when it was injected long before 
 the injection of the living and virulent microbe, but 
 also in the case of its simultaneous injection with the 
 microbe. Wooldridge thus furnishes the first in- 
 stance of the experimental cure of an infection by 
 means of vaccination. f 
 
 The interesting researches of Wooldridge, inter- 
 rupted by the premature death of this young savant, 
 have only recently been taken up again by Brieger,. 
 Kitasato, and Wassermann. But what is a little 
 singular, the authors having very well succeeded in 
 preparing by the method of Wooldridge chemical 
 vaccines against the different pathogenic microbes, 
 have been less fortunate with charbon. Perhaps 
 
 * Wooldridge, Arch. f. Anatomic und Physiologic, 
 Physiologische Abtheilung, 1888, p. 527. Wooldridge's first 
 publications date from 1887. 
 
 fThe fibrinogenous substances of Wooldridge are com- 
 pounds between the albuminoids and lecithin. He extracted 
 them from different fresh organs, and especially from the 
 thymus and testicles of animals. 
 
I2 9 
 
 these men have not been sufficiently persevering in 
 this direction, having the old prejudice as to the 
 radical distinction between the toxic diseases (tetanus, 
 cholera, diphtheria, typhoid fever) on the one hand, 
 and the septicaemias (charbon, rouget of swine, etc.) 
 on the other.* 
 
 Wooldridge's experiments on charbon have also 
 been continued by Hankin and Martin. f 
 
 Hankin has cultivated the anthrax bacillus in 
 Liebig's beef-extract with the addition of fibrin. 
 From these cultures he has isolated, by precipitating 
 them with alcohol, an albuminoid substance which he 
 calls albumose. This albumose has the property of 
 vaccinating against charbon, and even of curing this 
 disease. 
 
 We must, however, immediately add that Peter- 
 mann, who has repeated exactly the experiments of 
 Hankin, has arrived at entirely negative results. Be- 
 fore Petermann, Landi also obtained only negative 
 results. Martin| has cultivated the anthrax bacillus in 
 a solution of pure alkali-albumin. He has studied in 
 detail all the chemical products which form in cul- 
 
 *Brieger, Kitasato, and Wassermann, Zeitschrift f. 
 Hygiene, t. xii, fasc. 2. 
 
 f Hankin, British Medical Journal, Oct. 12, 1889, p. 
 810. 
 
 $ Sidney Martin, Proceedings of the Royal Society, 
 May, 1890. Annual Report of the Local Government Board, 
 
 London, 1889-1890, p. 235. 
 9000 
 
130 
 
 ture-broths. In accord with Kuehne, he has found 
 the three albumoses and the peptone which result 
 from the digestion of albuminoids by the proteolytic 
 ferments. He has also found an organic base. The 
 albumoses and the base are all toxic, especially the 
 latter. A toxic ptomaine was long since isolated 
 from anthracoid cultures by Hoffa,* who employed 
 three different methods for its extraction, and who 
 cultivated the anthrax bacterium on meat, or on eggs 
 mixed with bouillon. The ptomaine of Hoffa is, 
 however, very little toxic like that of Martin. 
 
 Lando Landi f has made some very interesting 
 researches on the substances produced by the anthra- 
 coid bacterium. He has found abase toxic to mice, of 
 the carbopyridic series, and some peculiar albumoses, 
 which may be obtained in a crystalline state; but his 
 attempts to vaccinate animals with all these products 
 have not succeeded. 
 
 Lastly, Christmas} seems to have obtained some 
 positive results. He vaccinates hares against char- 
 bon by two processes: First, by the diseased anthra- 
 coid organs crushed and emulsified in water, in which 
 the bacteria are killed by eucalyptus essence, and 
 the products then filtered through paper. In his 
 
 * Hoffa: Die Natur des Melzbrand Giftes. Wiesbaden, 
 1886. 
 
 f Lando Landi, C. R. de la Soc. de Biologic, July 25, 
 1892. 
 
 :{: Christmas, Ann de 1' Inst. Pasteur, 1891, p. 487. 
 
second process he makes anthrax cultures in a 
 medium composed of yolks of eggs, egg albumin, 
 and veal broth feebly alkaline, in equal parts. The 
 bacilli grow very well in this medium, but do not form 
 spores. After five or six days of incubation at 30 
 C , the cultures are diluted with water and passed 
 through the Chamberland filter. The filtered liquid 
 possesses feeble vaccinal properties. If the cultures 
 are prolonged beyond sixty-seven days, they become 
 very toxic and not at all vaccinant. These anthra- 
 coid poisons have not been studied by Christmas. 
 
 To sum up, we see that despite a considerable 
 number of researches the anthracoid poisons still 
 remain almost entirely unknown to us, and the little 
 that we do know is not yet established with certainty. 
 
 The poisons of glanders were first studied by 
 Finger.* He found that boiled cultures of the 
 glanders bacillus are toxic; but the phenomena which 
 they produce have no similarity to the symptoms of 
 farcy- infection. He has succeeded in some excep- 
 tional cases in conferring immunity on hares against 
 the living microbe by means of these boiled cultures. 
 Later, this question was again taken up by Bromberg,f 
 who has seen that even at 120 C. the toxic properties 
 
 * Finger, Ziegler's Beitrage zur Path. Anatom., 1889, t. 
 vi, p. 373- 
 
 f Bromberg, C. R. des Travaux de 1'Institut Veterinaire 
 de Kharkow, t. iii, i88g. 
 
132 
 
 of the cultures of the glanders bacilli are not com- 
 pletely destroyed. 
 
 Since then, the poisons of glanders have been 
 only studied from the special point of view of malleine 
 applied to the diagnosis of latent glanders, as we have 
 before seen (see page 78). 
 
 It is thus seen that our sources of knowledge re- 
 specting the poisons of glanders are scarit. 
 
CHAPTER XV. 
 
 PRACTICAL RESULTS OBTAINED BY THE STUDY 
 OF MICROBIAN TOXICOLOGY. 
 
 SUMMARY. Rapid Indications respecting the Other Bac- 
 terial Poisons Immunization as a Result of Toxi- 
 cological Studies. 
 
 We are obliged to stop here in our study of the 
 microbe posons. We must, however, before closing, 
 enumerate some of the special treatises on the sub- 
 ject which the student would do well to consult. 
 
 First, the memoirs of Charrin and of Bouchard's 
 laboratory on the pyocyanic poisons;* those of Mon- 
 fredi and Traversal and Roger on the poisons of 
 erysipelas; J of Chamberland and Roux on the septic 
 poison and that of symptomatic charbon; those of 
 Arloing on the poison of the bacillus heminecrobio- 
 
 * Charrin: The Pyocyanic Disease. Paris, 1889. Also 
 his opening article in the new Traite de Medecine, tome i. 
 
 f Monfredi and Traversa, in Giorn. Intern. Science 
 Mediche. 1888. 
 
 \ Roger, C. R. de la Soc. de Biol., July 4, i8qr. 
 
 Chamberland and Roux, Ann. de 1'Institut Pasteur, 
 1887, No. 12. Roux, Ann. de 1'Institut Pasteur. 1888, No 2. 
 
- 134 
 
 phylus; * those of Selander on hog-cholera; f those 
 of Hericourt and Rechit and Courmont and Dor on 
 the bacillus of the tuberculosis of birds.]; We must 
 also cite the researches of Schiff and of his successors 
 who have endeavored to solve the problem proposed 
 by Stich (see page 5). But we hasten to abridge- 
 all these details, which are very insignificant in com- 
 parison with the great principle of which it remains. 
 for us to to speak. We refer to immunization. 
 
 Thanks to the impetus which has been given the 
 last few years to the researches of microbian toxicol- 
 ogy, science has been able to discover and establish 
 definitively on solid bases a principle which is likely 
 to be of inestimable value in medicine. 
 
 According to the doctrine of immunization, it is 
 in our power to prevent and cure infectious diseases 
 by the serum of animals refractory and prepared. |[ 
 We have already seen how immunization is realized in 
 
 * Arloing: Les Virus. Paris, 1891. 
 
 f Selander: Contribution -to the Study of Swine-pest 
 (Ann. de 1'Institut Pasteur, 1890, No. 9). 
 
 t Hericourt and Richet, C. R. de la Soc. de Biol., 1891* 
 p. 470. Courmont and Dor, Arch, de Med. Exper., 1891, 
 No. 6. 
 
 Bouchard: Lemons sur les Auto-intoxications. Paris,. 
 1879. Roger: The Action of the Liver on Poisons. Paris, 1886. 
 
 || This preparation, we may remind the reader, is effected 
 by saturating these refractory animals with the toxic soluble 
 or vaccinant products of the corresponding pathogenic mi- 
 crobes. 
 
the case of tetanus. It has been applied to yet many 
 other diseases. First it has been established for fibrin- 
 ous pneumonia by the labors of Emmerich and Fovit- 
 sky and the two Klemperers.* It has been confirmed 
 by the researches of Archaroff.f Klemperer has given 
 the results of the application of immunization in the 
 treatment of forty cases of pneumonia in man.;); Im- 
 munization has also been applied to the rouget of hogs, 
 to the septicaemia of mice, to the disease caused by 
 Friedlander's microbe, and the pyocyanic disease. 
 More recently still, a way has been found for giving 
 immunity against typhoid fever, by Brieger, Kitasato, 
 and Wassermann,| and for robine by Ehrlich.^f Seeing 
 the great number of immunizations which have been 
 discovered in the short space of about eighteen months, 
 one might almost be warranted in concluding that 
 immunization is a general principle, and that, conse- 
 quently, all the infectious diseases are curable by this 
 method. On the other hand, the researches of Ehr- 
 
 * Emmerich and Fovitsky, Mtlnch. Medic. Woch. , August 
 11, 1891. G. and F. Klemperer, Berlin. Klin. Woch., August 
 24 and 31, 1891. 
 
 fArch. de Med. Exper., 1892, No. 4. 
 
 j Berlin. Klin. Woch., May, 1892. 
 
 Klemperer, Berlin. Klin. Woch., May, 1892. See also 
 Munch. Medic. Woch., 1892, Nos. 5 and 6. 
 
 || Brieger, Kitasato, and Wassermann, Zeitsch. f. Hy- 
 giene, t. xii, fasc. 2. 
 
 1 P. Ehrlich, loc. cit. 
 
- 136 - 
 
 lich, who applies immunization against the poisons of 
 plants, as abrine, ricine, and robine, demonstrate 
 peremptorily that it is against the soluble poisons 
 principally that immunization acts, and that by neu- 
 tralizing the soluble poison one becomes master of 
 the infection. And this gives support still further 
 to the principal thesis of my book: that the patho- 
 genic microbes are injurious only by their poisons 
 that infection is only intoxication by the bacterial 
 poisons. 
 
 Moreover, it was in the experiments with the 
 poisons of tetanus and diphtheria that immunization 
 was discovered. And it seems a little strange that 
 the discovery was not sooner made, for the studies on 
 the microbian poisons are less complex than the re- 
 searches on living bacteria. Thanks to the relative 
 simplicity of its problems, microbian toxicology has 
 been able to free itself of the first of the clogs of 
 preconceived ideas which impede and obstruct the 
 other domains of bacteriology. 
 
Antiseptics and Disinfectants 
 
 We desire to call attention to the following antiseptic and disinfectant 
 preparations: 
 
 Ethereal Antiseptic Soap (Johnston's) is a hydro-alcoholic solution 
 of Castile soap which was devised by an experienced nurse in the surgi- 
 cal clinic of the Jefferson Medical College. It may be made weak or 
 strong in antiseptic value by dissolving mercuric chloride in it in propor- 
 tions indicated in the case in hand. Since its introduction its use has 
 been extended to the treatment of parasitic affections with much success. 
 
 Antiseptic Liquid arrests decomposition and destroys noxious 
 gases that emanate from organic matter in sewers and elsewhere, and 
 may be used in cellars, barns, outhouses, and the sick-room. (Send for 
 Note 01 the Disinfectant of the Future, by Prof. Alfred L. Loomis ) 
 
 Antiseptic Tablets are convenient for the extemporaneous prepa- 
 ration of antiseptic solutions of definite strength for disinfectant purposes 
 and for antiseptic sprays. 
 
 Eucalyptus and Thymol Antiseptic is adapted for use as an anti- 
 septic internally, externally, hypodermatically, as a douche, a spray, by 
 atomization, and as a deodorant. Its application in surgery is unlimited. 
 It combines the antiseptic virtues of benzoic acid, boric acid, oil pepper- 
 mint, oil eucalyptus, oil wintergreen, oil thyme and thymol. 
 
 Disinfectant Powder possesses in a high degree disinfectant, ab- 
 sorbent, and antiseptic properties. It is admirably adapted for the dis- 
 infection of excreta in cholera, yellow fever, and typhoid fever, and in 
 all diseases in which such an agent is indicated for the purpose specified. 
 
 Labarraque's Solution we supply for the use of those who desire 
 to employ it for its local or internal antiseptic action. It may be diluted 
 to suit the indications. 
 
 Sulphur Candles thoroughly employed are effectual in the fumiga- 
 tion and disinfecting of rooms after infectious diseases. 
 
 We shall be pleased to forward, on request, any information desired con- 
 cerning these products. 
 
 PARKE, DAVIS & CO., 
 
 Detroit, New York, Kansas City, and Walkerville, Ont. 
 
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