TECHNICAL MYCOLOG) VOL. I.STANDARD WORKS BREWING: The Principles and Practice of. For the Use of Biodonts and Pracheat Mens By Waurun J Sees, MD, Third Walton terlaed by Aarzon R. Lama, Bdltor of the “‘Jonrail of the Institute of Browing.” In large Svo, handsome cloth, price 21s net « rvolnm of Browing Solotos, whioh has long boen awaited... « One of T= MOST COMPLUTZ IX CONTENTS and NOVEL IN ARRANGEMENT that has yet been pablished, . , Will command a largo aale."—The Breuers? Journal, “The appesrance of a work suoh aa this serves to remind us of the BNORMOUELY RAPID ADVANOnS made in our knowledge of the Scientific Principles underlying the Browing Prooasses. . . . Dr Sykes’ work will undonbtediy be of the GREATEST assisnsvon, not merely to Brower, but to all Obert and Biologists intrested in the Problems which the Fermentation Industries present,” —The Analyst. AN ELEMENTARY TEXT-BOOK OF BIOLOGY. ByJ.R. Amsworrs Davis, B.A. Professor of Biology, University College, Aberystwyth. 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FERMENTS, AND THEIR ACTIONS: a Text-Book on the Chemistry and Physics of Fermentative Changes. By Can ‘mErMER, Fb D., M.D. Translated by C. A, MrrowEnt, B.A, P10. loth, 7s, 8d. net «Buch a veritable mulium in parvo has never yet arpeaicd."”—Bresecrs’ Jours ab OILS, FATS, BUTTERS AND WAXES: Their Preparation and Properties, and the Manufacture therefrom of Candies, Soaps other Products. By 0. R ALDER Wnricet, D So, FRS. Secon Edition, Revised ©. A, Mitoumit, BA, FIO, FOS In large 8y0, handsome cloth, with 154 Illustrations, 268. net, “Dz, Warcut’s work will bo found ABSOLUTELY INDISPENSABLE by ever Obemfst. Tuma with information valuable abkke to the Anslyst and 1 ae a ¢ Analyst and the Teobnical *.* For other Works useful to Chemists snd Manufacturers, consult the Catalogue at the end of thie Volome LONDON: CHARLES GRIFFIN & CO, LMD,, RXBTER STREET, STRAND aNTECHNICAL MYCOLOGY THE UTILIZATION OF mrcRo-|__— ORGANISMS IN THE ARTS AND MANUFACTURES A PRACTICAL HANDBOOK ON FERMENTATION AND FERMENTATIVE PROOESSES FOR THE USE OF BREWERS AND DISTILLERS, ANALYSTS, TECHNICAL - AND AGRIOULTURAL CHEMISTS, PHARMACISTS, AND ALL INTERESTED IN THE INDUSTRIES DEPENDENT ON FERMENTATION By Da. FRANZ LAFAR ‘Professor of Fermentation-Phystology and Basteriology in the ‘Imperial Toshnieal High School, Vienna With an Introduction by Dn. EMIL CHR, HANSEN Prinetpal of the Caslaberg Laboratory, Copenhagen Translated by CHARLES T. C. SALTER IN TWO VOLUMES SCHIZOMYCETIC FERMENTATION WITH PLATE AND NINETY FIGURES IN THE TXT LONDON CHARLES GRIFFIN AND COMPANY, LIMITED EXETER STREET, STRAND 1gi0* [AU rights reserved . .660+ BB44F No3 Pamrep ay BALLANIYNE & COMPANY Lr, ‘Tavierook Stazer Covent GARDEN ‘LonpoxPREFACE Da. Larag has paid me the compliment of forwarding me a copy of tho first volume of his “Troanroa, Mycotoey,” with a request that I should write a preface to the work. A perusal of the book gives mo the impres- sion that its contents will in themselves be a sufficient recommendation, and ensure the success of the work through its own inherent value ; con- sequently, an Introduction by me isso far superfluous. Should, however, a fow words of mine be the means of helping to secure for the work of my young colleague readier introduction, here and there, than it would perhaps otherwise find, I shall be exceedingly pleased. ‘The First Volume treats of Baorerta, In a series of chapters we are shown the predominant réles—both useful and antagonistic—played by these organisms in Distillmg and Brewing; in the preparation of Wines and the Manufacture of Vinegar; in the Dawry; in Farming; in the preparation of Agricultural Fodder ; and in the manufacture of Tobacco and of Sugar. Then follows an account of the relation of Bacteria to sundry transformations occurring in Nature, particularly the important facts recently established mm connection with the combination of free nitrogen by bacterial agency, with the iron and sulphur bacteria and the bacteria of nitrification. Tt might be feared that, in a work aiming at objects so decidedly practical, the theoretical side of the subject would possibly be overlooked. This is, however, not the case in the present instance, as a glance at the Table of Contents will suffice to show. That the Anthor possesses a grasp of the historical development of the subject has already been evidenced m his previous treatises, and the same feature often appears in the present volume. In the majority of the Text-books and Manuals published in recent years, great confusion exists with regard to the appending of authors’ names to the Illustrations. In one and the same book, for example, we - meet with instances where the name of the anthor of the original work whence tbe copy has heen taken is giver—as 1 should be—and also v . «# PREFACE with other oases where the actual author is ignored, his name being ‘yeplaced by that of the compiler of some text-book from which the copy was * pbtained—i,, some one who himself has done nothing more than copy. “> gach a mode of procedure is in a high dogres calculated to produce a " gnisty conception of the actual circumstances in the mmnd of the reader, « ¢he more so because, as stated, no importance is attached to the occurrence. * Dp Lapar has, however, set vigorously to work to combat this bad habit by taking all his reproductions direct from the original sources, so that they afe clear and accurate representations of these originals. ~ ‘Tho subjects inclnded in the present work have been dealt with in a lany-sided manner, the Botanical as well as the Technical and Chemical aspects having been borne in mind, although preference has throughout ‘been accorded to the two latter. The style is flowing and clear, in many i” plates lively and picturesque, and I have read with interest even those % «portions wherein I am not at one with the opinions of the author, The ++ gttention devoted to the most recent developments of the subject gives a + special value to the book. Within the last two decades the stady of Microbiology has made gigantio strides, both in the pathological and the technical branches of the subject ; and just as investigations into the Physiology of the higher plants gave the first impetus to the establishment of Agricultural Expenmental Stations in all countmes, s0, in like manner, have the Physiology of Fermentation and ‘Technical Bacteriology called into existence, within the last few years, a number of Stations and Laboratories for the development of those branches of industry wherein micro-organisms play an important part. Formerly, Chemistry exercised on undisputed sway over the whole of this realm, but now Biology has won for itself a co-ordinate position therein—a fact which ib bow being recognised (although not yet to an adequate extent) in the , filling of professorships at the various Technical High Schools. An army ‘de of eager workers has arisen, new technical journals have sprung into existence, and « great number of treatises and books are published on the subject every year, However cheering this may be in itself, the fact can- » ° not be gamsaid that a portion (unhappily much too large) of these publica- tions ought properly never to have geen the hght. It is trae that an intimate connection with practical conditions sets fresh tasks before the _ investigator, and exerts on the whole a sufficiently stimulating influence; but, on the other hand, the same circumstance gives rise to the danger of diverging intg by-paths, and neglecting the strict scientific conditions of s .PREFACE vii investigation. Since these Stations and Laboratories are, as a rule, main- tained by the cirele of practical men for whom they work, the investigators appointed thereto are often subjected to regrettable pressure, Even though, otherwise, a certain amount of freedom is allowed them in these institutions, they labour under the great difficulty of being obliged —whilst engaged in the task of scientific investigation—to be ready at any moment to give assistance—coupled with analyses and any wished-for disclosures— to the parties interested. Still further difficulties arise when practical men foolishly intermoddle in scientific investigations, and especially when results that shall be immediately available for practical utilisation are impatiently demanded—results which, however, are only attainable by scientific investigation, and cannot be forced on at pleasure. Under circumstances of this nature it requires great strength of character not to give way to outside pressure, and many examples are met with in the literature of the subject where this firmness has been lacking. The result of these vexed relations between Scientists and practical men has been to call into existence quasi-scientific literature by which neither Science nor Practice has benefited—a result which every one who has the healthy development of this sulyect at heart must greatly deplore and endeavour to improve according to his ability. These conditions are, however, in existence, and we must take them into account. Amongst the chaff which occnpies a large part of the aforesaid technical journals, there is, nevertheless, come really good grain to be found, and he who undertakes to write a work on Technical Mycology must not content himself with gathering from purely scientific sources alone, but must, at the same time, work through the technical journals as well. This (by no means easy) task has been accomplished by Dr. Larar with commendable discernment and ability. In the last few years, certainly, we have had various Text-books and Manuela giving a summary of larger or smaller sections of Technical Microbiology ; none of them, however, has treated the whole of this exten- sive field from so comprehensive a point of view. To prepare a work like the present requires not only many-sided discernment, but also enthusiasm for the task, combined with courage and endurance—properties with which the book shows the author to be endowed. The work will be welcomed, not only by those for whom it 18 primarily intended—viz., Technical Chemists, Chemists dealing with food-stuffs, fermentation, and agriculture,vu FREAUL Pharmacists, and Agriculturiste—but many a professor also will derive benefit from its pages for his lectures and rescarches. In this respect the copious bibliographical references will also be of good service, In the present volume we have unfortunstely only the numbers of the references, it being intended that the Bibliography shall be published as an Appendix to the second volume. This inoreases the desirability of the early appearance of the latter. Tho Publishers have prodaced the work in a handsome and substantial manner, and in this respect also the impression produced is of the best. EMIL CHR. HANSEN, Cantaenc Laponarory, Corenaacen, September 1896.LIBRARY a O. Ca TABLE OF CONTENTS. INTRODUCTION. I—tTsxz Taxory or SponTaNEOUS GENERATION. PAGE TAGE § 1. Definition 1 Foundation of the Science of Antl- § 2 Discovery of Fermontativs O1- septios by Schwann, as ganiems 2/8 Schroder and Dusch 5 § 3 Noodham's Domonsiration in favoui § Examination of the of "Generatio qmvooa” . . 3 . 6 § 4. Spallanseni's Bxperments 4 | § 8 péotaurd's Misrosyine Theory 7 § § Frans Schultze’s Expeument 4 | § 9 Spontaneous Generation only Uu- proven, not Imposible . . 8 IL—Tasxorrs or FERMENTation. § 10. The Alchemists—Stahl's Theory § 17.0 Nigel's Physico-Moleoular of Fermentation =. =. 10 Theory : 16 § 1 Gay-Lusags Opinion "| 104 § 18 The Bosymes and Mi. Traube's § 12 Ongniord-Latour’s Vitalistio Ferment Theory y Theory zi 11 | § 19. Ganeral Detintion of ‘Formenta- § 13 Th. Schwann’s Researches +2 Fi 18 1g Fr Kutsing’s General Theory 15 | § 20 Sovatled Spontaneous Ferienta: i 15, Lacbg’s Decompoation Theory 14 thon of Sweet Fruits. . 19 § 36. Pasteur's Thoory . «x6 | § at. Decompositions offeted by Light and Ar. 19 Ti.—Te Orcanisus or FarMxntatioy, § 22 The Postion in the Botantoal § 25 Assmilation of Carbon Dioxide System. ar ‘without the Aid of Chlorophyll. 23 § 23, Clasatiention of the Fung... 22 | § 26 Saprophytes ond Parasites 23 § 24. Bchizophyte « toa o- DIVISION I. SCHIZOMYOBTIC FERMENTATION Section I.—General Morphology and Physiology of the Sehizomycetes. Cnarrer I—Form anp Drwmnsrons. £25 Home ot Grown «+ + 35 | 9 29. Mutabiltty of Form + 28, Dimensions of Bacteria. + 27 | § 30. Involution Forms =. 0.28 Ee . . .x CONTENTS Onaprer I].—SrrvcruzE AND a oF THE BACTERIAL Le race § 31. Chemical Composition of the Cell Wall ee ek § 32. Optical Properties of the Cell Wall 30 § 33. Zooglosa Formation =... 3r $34. Plasmolysis . . sss 32 435. Structure of the Cell Contents 36. Blomentar Bacterial Cell“. PAGE Composition of the § 37 Quantitative and Qualitative Se- lective Power - . + Onaprer III.—Power or Inpermnpent Movament rx Bacrenta. § 38. Molecular Movement and Looo- ‘motion on cee 7 § 39. The Flegella or Oia +s 37 Onarrn IV.—Vearrative § 42. Division in One Direption - 43 § 4. Division in Two Direotions + 43 § 44 Division in Three Directions ©. 43 Onaprea V.—Tue Permavenr (Rernopucrivs) Fonws on Srones § 48. Formation of Endospores . 46 § 49. Alterations in the Form of the Mother-Cell . . «+ 48 § 50. Number of Spoies 148 § 51. Form and Size of the Spores’ +49 § 40. Histology of the Cin. ES Ohemotarn ee ‘Repropvorion sy Fission. § 45, Form of the Daughter-Cells « § 46. Divisionof the Nuclews + § 47. Rate of Reproduction . § 52 Conditions Influencing Spore For- mation + § 53 Reusting Power of the Endospo.os § 54 Bohaviour of the Bndoepores to- wards Dyess § 55. Arthrospoies ‘ Cuapran VI.—Tun Gzrumartion or ta8 Enpospore. $56 Bret Type: see 52 § 57. Second Types 5 Ss ge $58. Murd Types LL § 59 Importance of this Process in the Classification of Bacteria « Section Il.—General Biology and Classification of Bacteria. Onarrmn VII.—Te Baoreria uNDER THE INFLUENCE oF PrysicaL AGENoTES, § 60, Infuence of Electricity § 61, Influence of ‘Temperature—Cold and Heat-loving Bactena = e . § 62 Influence of Light—Self-Purifica- tion of Biyers § 63 Influence of Mechameal Shock, Gravity, and Gaseous Pressure 33 35 36 44 44 “4 49 49 50 3 35 63CONTENTS a Ouarran VIII.—Baorenia mv raer RELATION 10 oN ANOTHER. Page . rAgE § 64. Symbiosis, Metabiosis, Anta- § 65. Mixed Cultures . 1°. + 67 ro + 6 . Onarran I[X.—Otassrvrcation or 1H= BacTERIA 7. Cohn’s Classification . 69 om f 68. Biliroth’s Ooceobacteria Seption. 70 | § 70 Pathogeme, "Chromogenic, and Zymogenio Bacto. + «73 96. First Attempt by OF. Muller . 69 $6 n § Seetion III.—Principles of Sterilisation and Pure Cultivation. Onaprer X.—Mernops or SrermisaTion. § 71.Bterihamg . - 715 bon dioxide, chlorine and chlo- § 72. Froang the Air from Germs % nide of lime, ozone and hydro- § 73 Filtration of Dnnkung Water. . 77 gen peroxide, milk of ime. 8 4 74 The Bacterium Filter m the Ser- § 80 Organio Antiseptios. Phono, ‘vice of Enzymology. 2B resol, oreolin, Iysol, sapocarbol, frp Ber Rute, os 2B solveol, solutol, eaprol, salioylic § 76 Dostroymg Germs by Dry Heat "79 acid, antinonn, ethyl aleohol, § 77. Destioying Germs by Moist Heat 80 ethylether, formaldeliyde, iodo- 278 tutermttent Sterintion » Be form, chloioform, benwore acid 87 § 79. Mineral Antiseptics * Coriosive § 8t The Combmed Method of Sterili- sublimate, sulphuious acid, cat- sation se OF Oxaprer XI.—Mazrnops or Pure Ovurore. § 82 Nutrient Solutions 94 | §85 Kooh’s Plato Cultures—Streak § 83 Tho Dilution Method and Frac and Punetue Cultures—Pre- tional Culuvation . 96 serving Cultues — Auxano- § 84 Liqueflable Sod Media - 99 graphy ee ee 108 Seetion IV —Chromogenie, Photogenic, and Thermogenie Bacteria. Cuapren XII —Ouromorarous Baoterta propucixa Rep anp YELLOW Corouzine Marrers § 86 Coloured and Colouring Bucteria 105 | § 89 Red Coloration m Milh, Cheese, § 87 Mictococous Piodigioss . «108 and Stook-fish ; 108 $88 Lipochomes . . + + 107| § 90.Bactais producing Yellow Colouring Matters. 109au CONTENTS Oxaprar XIII.—Punrig Bacrerta AND THEIR BEHAVIOUR TOWARDS Lrenr. . AcE pace 9t. Their Morphology + rir | § 93 Assimil oo ition and Oxygen Elimi- 92, Influence of the “Individual mation see 113 Colours of the Spectrum =. 112 Onarren XIV.—Onromoranovs Bacrerta propvorna Biv, GReny, AND ‘Viount Couourrsa Marress. $94 Blue Coloration of Milk =. 9. 115 =Bacteria producing Violet § 95. Blue Coloration in Ohease . 9.117 Pigments =. it § 96. Tho Fermentation of Indigo. 1x9 | § 98 Gieen Bacteria—Green Coloia- § 97. Varieties of Baotllus Pyooyanous tion of Cheese 2s Tae Osarten XV.—Puoroaenio Bacrenra. $ 99, TheGenus Photobaotenum . 123 { 100. The Food Requirements of Phos- phorescent Bacteria + + 124 § 01. The Lummons Bactens as Tests for Enzymes. «5. 125 § x02, The Phosphorescenta . + 125 Onarren XVI.—Taermogunto Baorsni, § 103. Spontaneous Combustion. wy § 105 The Fermentation of Tobacco 128 § 104. Spontaneous Heating of Hops i: § 106 The Preparation of Burnt Hay. 129 tion V.—The Heat-Resisting Bacteria : Their Place in Nature and their Importance in the Fermentation and Food-Stuff Industries. Quapren XVII.—Bacmtvs Supriis anp rms ConcEnuns, { 107. Roberts’ Heat Method 131 | § 110. The Potato Baoillt 134 J Yoh Morphology of Bacillus Bubbhne 132 | § 111. Bagilas Frtslans—Prodaction 109, Influence of thé Mode of Nutri- ‘of Alcohol by Fission Fungi 136 . tion on the Form of Growth . 133 | § 112. Bacterial Content of the Sol . 137 . Oxarren XVIII.—Borvaro Acip Fermentation AND ALLIED Decomposition Processes § 103 Anabiobions ne 139 | § 117. The Equation of Butyne Fei- 114. Methods of Oultrvating Ause- mentation. + uy robio Bacteria. + 140 | § 118 The Fermentation of Cellulose 148 9 115, Clostridmm Butyricum (Praz- § 119. The “Retting” of Flex and ‘mowaki) and Bacillus Buty- Hemp. + 15t mous (Hueppe) . «= «143 | § 120 The Rancidity of Fatu 132 § 116. The Genus Gianulobacter . 145 .CONTENTS PAon § rar. Dit-and Germ-Content in Mille "154 § 122 The Part played by Milk a5 0 ‘Carrier of Infections Diseases. 155 123. Boiling Milk =. y, 156 x24. The Soxhlet Bottle . 157 125, Germ-Oontent of Milk treated by the Soxhlet Method. 158 Onarraz X1X—Twm Pauservation or Mrux, § 126, The Method of Neuhauss, Gron- wald, and Ochmann ". § 127, The Content of Pathogenic Germs 1m Varlous Daury Pro- duots—Preserving § Mrlk for Analysis. § 128 Condensed Mik 1)! Osaprun XX —Tus Presenvation or Muar, Eoas, Vacerancus, axp Faurr { 129. Storage in Gold Ohambers —, 63) § 133."Demtooating and _Preserying § 130 Dried Meat and Salted Meat ‘164 SM Vegetables end Frat Gore, §13r Smoked Meats and Corned Boot 165 centrated Wine-Must =. ¥32 Proserving Hggs . ., (166 Section VI.—Lactie Fermentation and Allied Decompositions. Onarren XXI.—Gunznar Onansorunigros, § 134. Discovery of the Lacho Acid Bacteria : $ 135. Bacterium "Inotis Lister, and Bacillus aeidi Inctio Husppe , 171 170 § 136. The Equation of Lacts Fermen- toon. , ke Ouarrun XXII.—Tau Puopvotion or Oprioatzy Aoriva Oxcanro Compounns By 74 175 7 137. Isomers of Lactio Acid : 138. The Isomerio Tartaric Acids 139. The Division of the Ravemfe Compounds . . . Farmentation. $ 140. The Production of the Stereo- aomerlo Lactic Acids. Onarrer XXIII.—Tam Arrmomn Sounmne or Opnam, 141. The Acid Generator. 180 142. The Aroma of Butter. | § 143. Defeots in Butter. . Onarrer XXIV.—Tae Ooacu.ation (Ovrpime) or Minx. 144 Aad Curdling and Ronnet Ourdhing 1 § 145 Characteristics and Activity of Lab 185 § 146. Lab-Producing Bacteria... § 147, Onseare LL xiii rage 159 160 1 167 172 178 182 186 186 ~?xiv CONTENTS Cuarren XXV.—Laor1o Ac Bacruri iw Distiuiinc, Barwine, AND VINIFIOATION. Paar Page $ 148 The Spontaneons Acidificatron $150 Effont's Hydrofuorie fAcd of Distillery Yenst-Mash . 188 Method eS . 19T £ 149. Artificial Sourlog by the Ard of $151. The Laotio " Acidificaon Pure Cultures of Laotio Acid (*Zickendwerden”) of Wine. 193, Bactoria, Bacillus acidificans § 152. The “Turning” of Beer 194 Tongionumus «189 | § 153) White Boer, Lambie, Gunger- er a terest 1195) Cuarren XXVI.—Tae Lacrio Acm Baorsera 1n THE PREPARATION or Fopper. §154BrownHay . . . . 199 | §156,SourFodder . . « . 202 $155. Bweot Bnailage | 1 | 200 Onarter XXVII.—Tne Pant Pravep sy Baormara mv Tannine, § 157, The Fermentation of the Plump- $ 158. The Souring of Bark Liquor , 205 ingBonk. =... + 204 Section VII.—The {ortation of Mucus and Allied Phenomena of Decomposition. Onarrer XXVIII.—Tue Imrontanoa or BacrsRta IN THB Manvractury or Suaar. § 159. The Zooglea of Leuconostoc senterioides = - , =. 208 § 160 Physiology of Leuconostoo 210 § 161. Muoinous Fermentation and In- version . oe = 212 Onapren XXIX.—Rormuss in Mux, Wave, Bune, anp orman Liquipa. § 162. Ropy or Visoous Milk + 215 | § 166, Ropiness in Wort and Beer. 219 § 163. Ropy Whey ond Thick Mitk . 276 | § 167. So-called Sarcima Turbidity in 164. Ropinessin Wine.) . | 217 (Beoer peasy ee eatect ese a 365. Ropiness in Infusions §= || 218 Section VIII.—Decompositions and Transformations of Organie Nitrogenous Compounds. Onarren XXX.—Tue Poenomena or Puragracrion. § 168. The Degradation of the Albu- nitnfcat on—Nitrio Fermen- minods. « + 233 tation of Molasses. + 235 § 169. The Putrefactrve Bacteria 226 | § 174. Lows of Colour Umschlagen) m § 170. Proteolytio Eosymes. . —. 230 ‘Wine 339 $171. Ptomaines and Leucomarnes | 232 Fermentation of Urgamo Acids x72. The Albuminous Powons .. 234 by Sohizomycetes. =. 240 -§ 173. Laberation of Nitrogen, an De- ‘Mannitio Fermentation of Wine 241 . . .ogy is the Science of the Character t Fermentative Organisms, Fermentative organism 1s the name given to any minute being of vegetable nature capable of exciting fermentation Whether any given minute organism 1s to be considered as u “fermentative organism ” or not depends, there- fore, on the answer to the question. “Does it possess the power of causing , or, 1n other words, to establish the common Ad PkedeiGes classified under that general title, ‘Thus is, however, as will soon be apparent, no hght task, and the probabihty of our attempts being crowned by a successful result will be greater if we limit the scope of the question at the outset, and for the moment consider tho term fermentation ” as applying merely to those phenomena with which 11s nasociated in colloqmal Janguage, vis., the conversion of must into wine, wort into beer, wine into vinegar, and fresh milk into sour, dic, To these may also be added the process of putrefaction, Adhering to tion of the term, let us follow m imagination the path of investigati 28 led to the knowledge that all the above-named phenomena are ie by the activity of minute living organisms, and constitute a manifestation of their vitalty , that fermentation and. putrefacton are, in short, not purely chemuoal molecular transformationa, but physiolograt processes, Moreover, history shall be our instructor, and lead us on further, to the com- prehension of those other processes, which, for the present, we assume as standin, without the pale of the term “fermentation,” but which, neverthelees, should actually be included therem. Such processes are, inter alia, the transformation of ammonma into mitre acid, occurring in the soil of pur fields, the decomposstign i 1 42 THE THEORY OF SPONTANEOUS GENERATION and dissolution of dead vegetable matter; the ripening of cheese , the formatio of bog (tron) ore, da, do, § 2.—Discovery of Fermentative Organisms. ‘The organisms taking part in the processes of fermentation are so minut that only a few can be detected, and that very imperfectly, by the unassisted ey The term microbe, introduced into the vocabulary of science by O Smpmicr (I.) 1n 1878, belongs to them of right. Their examination could not be carried o anterior to the invention of appliances for observing minute bodies under hig powers of magmfication, and Therefore the inventors of the microscope deser ‘to be held in grateful remembrance in the domain of fermentation, These wei Hans and Zacharias Janssen, father and son, spectacle-grinders, of Maddelbur; in Holland, who, about the year 1590, constructed a combination of lense which, although, of course, very amperfect when compared with the instramer of the present day, must be regarded as the first compound microscope made ‘Nevertheless, however great this step undoubtedly was, both from a theoretic and practical pont of view, and however fruitful 1t proved in results, seeing the at rendered possible Zater discoveries in the world of the “ infinitely little,” an especially of the fermentative organisms ; still the fact remains that the ir: fundamental observations were made, not with the compound, but with th simple miroscope, which then, as now, was ttle more than a magnifying glae or bi-convex glass lens, ‘The honour of having discovered the presence of extremely small and hithert undetected organisms in putrescent ad fermenting hquids belongs to anothe native of Holland, by name Antony van Leguwannonr. Born at Delftin 1637 he acquired during his apprenticeship to a hen or cloth merchant in Amsterdar some ékill im grinding small glass lenses Of this skill he made further use afte his final return to his native town, and succeeded in producing lenses capable o magnifying from 4o to roo, and even to 150 times. Wath these he examine various minute objects, and frequently, amongst others, all lands of vegetabl infusions in a state of decomposition. He discovered therein sundry extremel, small creatures, many of them capable of motion, which he therefore regarded animals, and named from their habitat infusoria. He died in1723 The moder: world has entitled him “the father of micrography,” 2.6. that science which treat of the most minute forms of life. This newly-discovered field of research was at first regarded by Leeuwen hoek’s successors from an almost exclusively medical standpoint, as it 16 1 natural instinct in man to try and maintein health and to prevent disease A. that particular period, too, a special impetus was given to the stindy of medicin' by the ravages of the plague, which only too frequently pursued 1is destructivi course throughout Europe. On the other hand, the study of the phenomena of fermentation derivec litle or no benefit from Leeuwenhoek’s discovery. The first investigator whom we meet with in this domam is the Viennese physician, Marcus Antoniu: Plenciz, who in his work “ Opera medieo-physwa,” issued m 1762, applied the results of Leeuwenhoek’s discoveries, not only to the field of medicine, but alsc to that of fermentation and putrefaction In the latter connection he arriveo at the noteworthy conclusion that ‘a body undergoes putrefaction when the germs of vermicular creatures begin to develop and multiply; because these ammals excrete numerous precipitations consisting of volatile salts, by which the liquids are rendered turbid and malodorous.” 4 Tho Roman numerals given in brackets after the names of investigators refer to the Bibliographical References formifg an appendix to the second volumeBeOF “GENERATIO ZQUIVOCA” 3 ‘acquaintance with these minute creatures may 8 who succeeded Plencw, and however useful, Hlow, ight be to observers the processes of de- Gluced, these questions were nevertheless forced ound by another, namely, the ongin of these However have been to from a practia composition wh temporarily inte “tht minute organisms. - How do the minsite Some opined hrf (and therefore liquid in question, (generateo apontancé}y tion), Or, whilst pro (heterogeneous), or OF genérato equivasiik: noted, have the saute: The party oppe from a lifeless to soria” are detested & their exstence is’ ‘Which view ind eighteenth contury; 8 80 coprously developed in anfurions originate t -organisms were produced from certain unorganised f substances—chemical componnds—present in the ition being therefore considered as spontaneous ising from elementary substances (primary genera- from elementary substances, as diifering therefrom. thereto (oquivocal), hence the name Heterogenesis which terms, as well as that immediately to bo , on the other hand, the possibilty of a transition ition (abiogenesis), and asserted that when “infa- sion, a liquid or matter undergoing decomposition, @ germs present therein, this point there arose, about the middle of the med one of the hveliest disputes agitating the domain of natural that period, and which, after occupying the most earnest attention of Buooosaive gonerations of scientists, only termmated, after numerous duoiy out the middle of the present century. Fiom among the nur ors who took part in this controveray, mention, can here be magi Needham on the one side and Spallanzani on the other being em § 8.—Needham’s Demonstration in Favour of “Generatio Hquivoca.”” ‘The most energetio ohmmpion of the theory of spontaneous generation was the Enghsh divine Namygae (L). This theory was in existence long before hus time, and ha@.Sid'séhowned supporters—among them the chemiet Van Helmont, who proposed @ method for producing artificial mice—but until then had not progressed beyond the stage of indefinite assertion aud unfounded hypotheses. ‘The cause of the extraordinary supports and approval accorded to the assumptions put forward by the English divine 1s, on the other hand, attributable to the novel mavner in which he arrived at bis theory (pub- lished in 1745), viz, not by untenable hypotheses, but by well-directed expert- ments, ‘Ho set to work, for example, in the following manner An aqueous meat extract was boiled for a short time in a flask, which was then made air-tight and left to stand for several days or weeks When opened at the end of this time, the contents proved to be plentifully infested with “infusoria,” from which Needham concluded that as the “eggs” originally present in the liquid were lulled by the boiling and the entry of fresh ones from the outaide wes precluded, therefore the living infusoria discovered in the liquid on re-opening the fiask must have originated spontaneously, not from eggs (germs), but from the lifeless constituents of the quid. ‘The great impression produced on his contemporaries by these statements can be appreciated by reference, for instance, to Buffon’s work on the “System of Generation.” .4 ‘THE THEORY OF SPONTANEOUS GENERATION § 4,—Spallanzani’s Experiments. OF the two hypotheses formmg the basis of Needham’s deduction, the accuriey of the second, ie. that relating to the exclusion of outside germs, was examined first Some twenty years after the appearance of the Enghsh theory, the Abbé Srauuanzanz (I) published a dissertation in which he combated the doctrme of spontaneous generation In ths work the Itahan divine detailed the expesments which had Jed him to the conclusion that a development of the animalcule m question, in an infusion maintained at boiling-pomt for three- quaters of an hour, was only possible provided au, which had not been pre- viously exposed to the influence ot fire, had been adnitted This position was also maintained by Spallanzani in a second treatise (IT ) Nevertheless, the suppotters of the spontaneous generation theory were still far from regarding thew cause as lost. ‘They characterised these experiments as, aneonelusive, sneo (so they said) “by the immoderate heat Spallanzam chose to employ, the ar in the vessel 18 so unfavourably changed, and rendeed so unsuitable for the maintenance of life, that it 1s no occasion for surprise that all development was lacking” ‘This objection was cw tly rejected by Spallanzani, but an experimental confutation was only ariived at much later. The next step an this direction was accomplished in 1836 by— § 5,—Franz Schultze’s Experiment, In order to avoid under-estimating the value of the very short treatise (I) pubhshed by this mvestigator, 1egmd must be had to the influence attained by Obemstry in all branches of natural science during the sixty years that had elapsed since Spallanzan’s demonstration, an mfuence which will be eluci- dated, in so far as xt refers to the theny of Fermentation, 1n subsequent sections The idea that ordmary ar acts as an mducer of fermentation or potrefactaen, by reason of its content of livmg germs was first called into extstenee "by Yio 1—Franz Sobultze's Bxpertmeut Schultze. He desembed his experi- ment os follows “T filled a glass fask half full of distilled water (Fig’ 1) with which I had mixed various animal and vegetable substances, and dlosed at with a sound cork, through which wee passed two tight-Attmg glass tubes bent to elbow jamts, I next placed xt in a sandbath and applied ‘heat until the water boiled briskly, eo that all parts were exposed to a temperature of 100° 0. Whilst the hot water vapour was still issuing from the two tubes, I attached to the end of each an apparatus employed by chemists, im tho course of organic analyses, for the absorption of carbon dionde. ‘That on the left-hand side was filled with concentrated sulphuric acid, the other with a solution of potassium hydroxide” After cooling the apparatus, aur was drawn through twice every day durng the ensuing two months, in such a manner that xt had to pass throigh the sulphuric acid before entering the flask. ‘The reoulte confirmed the expectations gf the investigator, the contents of the flask when‘THE LABOURS OF SCHRODER AND DUSCH 5 ed being found free from living orgamums, which, however, soon made then \wance when the open flask was freely exposed to the air. This proved that ous exposure to the influence of fire 1s not an essential condition for ving air of the power of inducing fermentation or putrefaction hres years later, Taropor Somvann (II entered the fl as an opponent e theovy of apontaneous generation Of his labours in this direction, : modification of the Schultze experiment, consisting chiefly in the substi- 1 of & heated metal tube for the bulb tubes (see Fig. 2), occupies merely dary position More ‘ant in the attack on theory of the spon- y of the phenomena moentation was the ishment by him of et that a resort to is unnecessary in the ation of such decom- on, but that the same em be attained by ition of some toxic nee to the hqud nentation is arrested y influence proved eof killing ‘the fang, oma uly by heat, potas: Fro, a—~Theodor Selavann's Rxpertment, avoniate, &e.” Ho horefore, the founder of the science of antiseptics, Concerning Ins nental researches in the narrower field of alcoholic fermentation, mention ymade in a, subsequent chapter. ondherents of apontaneous generation applied to Schwann’s method of ing the nix the same objection (referred to above) which they had previously against Spallanzani ‘They gid not even consider themselves confuted by sults of Selvultze’s experiment, but asserted that here also the treatment of ‘,nlthough by no means so violent, unfavourably modified 1t@ composition, futation of this doubt was only accomplished after a lapse of seventeen and that by § 6.—The Labours of Schréder and Dusch (1). tignted by the researches of Loewel, who found that ordmary air could be xl of its property of inducing crystallisation ma supersaturted solution um sulphate by filtration through cotton-wool, the two anvestigntors above modified, in 1853, the arrangement of Schultze’s expenment, by ig the incoming air to pass through a glass tube packed with cotton-wool ontoring the flask It was found that by means of this (decidedly not at”) trentment the arr also lost ita power of causing decomposition and mation of minute orgamsms in extracts which would remain wochanged i was excluded. »importance of this demonstration must not, however, be over-estimated, nly proves the presence m the ar of a “something” capable of giving hying creatures in mammate nutrient media, and of exciting substantive 4 (fermentation and putrefaction) therein, Concermng the nature of this ‘something,” the experimenters could give no satisfactory account, they ft 16 an open question whether the something was gaseous or not It6 THE THEORY OF SPONTANEOUS GENERATION may be considered that they were unduly diffident, since the action of the cotton-wool filter proves that this something must necessarily be = solid body and not agas. But, on the other hand, both investigators could pomt to experi- mente wherein the previously boiled test quid afterwards underwent decom- position, notwithstanding the fact that all the air which was allowed access to 16 had been filtered through cotton-wool Milk they had, m their first treatise, recognised as such a liquid, and to this were added, m a second communication by Sonnopen (I.), yolk of egg, meat, and meat broth, in all of which cases the filtration of the air proved useless. ‘This Jed Schroder to separate the pheno- mena of decompostion—characterised as fermentation and putrefaction—into two groups’ the one, which he designated “voluntary decomposition,” requring only oxygen for its meeption, whilst the other, ¢g. the fermentation ot wort, required, in addition, the collaboration of thut unknown constituent of the air, which could be destroyed by fite or arrested by a cotton-wool filter “ Whether this active substance should be regarded as germs floating in the air, or as some Iutherto unknown chemical substance modified by high temperature and sepa- rated andi fixed by the influence of contact with the cotton fibres, must remain undecided ” Glancing back for a moment at the work of Schultze, one would be only too readily disposed to consider the results of Schroder and Dusch’s experiment as a retrogade step, since they not only did not afford us any further information beyond that established by Schultze as to the nature of the germs in the air, but also called in question the accuracy of Schultze’s results, And, in fact, repeti- tions of the Schultze experiment by many other workers, with various modifi- cations, especially with regard to the kind of test hquid employed, confirmed the results of Schroder and Dusch. In numerous mstances decomposition ensued, even in the boiled liquid, when purified sur (filtered or heated to redness) alone was admitted, whilst am other cases, under preowely similar conditions, the boiled sample remained unaltered for any length of time. Thus the state of the question at the commencement of the sixth decade was jnat about as far advanced as at the begining of the century, and the adherents of the spon- taneous generation theory were more certain of trumph than ever. § 7. The Examination of this Theory by Pasteur. However, the day of refutation was close at hand, though the proof was not obtained by the methods which had generally been favoured hitherto, but which had Jed to no definite issue. Expermenters had so concentrated their attention on keeping the air admitted to the boiled hqud perfectly free from active gorms, that 1t had not occurred to any one to ask if the sterilisation of the hquid could not be equally ensured by sumply boing it, either momentarily or continuously for a short tame. Reasoning from the fact that all known forms of created life (ammal as well as vegetable) were incapable of resting the temperatare of bailing water, even when exposed thereto for merely a short time, the conclusion was arnved at that the same effect was produced on the small germs m question. It was therefore conmdered, humanly speaking, certain that every hquid could be rendered free from active germs by boiling for a short time, This was agreed to both by those who accepted and by those who rejected the doctrne of spon~ taneous generation Still such belief was based on a mere assumption, as On. Boxwzr (I), 2 contemporary of Spallanzany’s, imphed when he mserted the following query in hus work opposing the theory of spontaneous generation ° “Tat, then, certain that there exst no ammals or eggs capable of supporting &BECHAMP’S MICROZYME THEORY 7 temperature equal to that of hot ashes without losing their life or reproductive wer 2” Po" Pastour called to mind this doubt of Bonnet's when he began to subjeot the theory of spontaneous generation to experimental examination in response to the offer made m 1860 by the Paris Academy of Science of a prize for “an attempt, by means of euitable experiment, to throw new light on the question of spontaneous generation.” From the report of his researches, which appeared early in the year 1862, in the form of a comprehensive treatise (I.), well deserving perusal, only the most important result can be referred to here viz., the demonstration of the posmbihty, by the sasistance of sufficiently prolonged heating at an adequately high temperature, of sterilising (1c. freaing from living germs) any substance whatsoever, and of the fact that a sample so sterilised will not subsequently undergo decomposition, but will remain unaltered so long as care is taken to prevent the access of germs from the external arr. ‘The objection raised by the heterogemssts, viz., that decomposition is pre- vented by the atrong heating having rendered the sample unsuitable for the production of germs, can be easily disposed of by inoculating the liqmd with a few germs, these will be found to develop rapidly and luxurntly. ‘The sub- stantiality of these germs was demonstrated by Pasteur ine very beautiful experiment, for which he employed a culture vessel similar to that described by H. Hoffmann (1.) in 1860, and now generally known by the name of Pasteur flack, a glass flask (fitted with o tubulus at the ade for facilitatmng inoculation) the neck of which 18 drawn out small and bent twice like a awan’a neck ‘Tho external aur 1s obged, in order to gain access to the contents of the bottle, to pass through this neck, and as the direction of movement 1s changed at the first bend, all the germs are deposited there, ‘Thus was laad the foundation on which the edifice of Fermentation Physiol was gradually raised, ‘The possession of perfectly sterile culture media, and the power of protecting them from the intrusion of unauthorised germs,1s a eine gud ‘non for a successful and rehable study of the organisms of fermentation. § 8.—Béchamp’s Microzyme Theory. Pasteur’s investigation and elucdation of the causes of the tenacity of life exhibited by many germs thenceforward ocoupied the earnest attention of amyeologusts, and finally led to the acknowledgment that this power of resistance is possessed by the reproductive organs known as spores, ‘The morphology and physiology of these organs forms the subject of §§ 48 to 55 At present, the only point to be emphasised 1s that when these hfe-retentive organs are once kalled, no spontaneous development of germs can occur in the liqmd harbourng them; hence such hquid will remain sterile until it 18 artificially re-mmoculated. Tt might be supposed that the adherents of the doctrme of spontaneous generation would have responded to these demonstrations by abandoning ther Previous attitude of oppomtion This, however, they did not do, they merely changed the field of combat without altermg their opmion As they could no longer maintain that orgamsed creatures could be spontaneously derived from unorganised substances, they contended that the dead cells had the power of liberating organised living matter capable of development into the various species - Tn a subsequent paragraph we shall Jearn that m the cell contents of most fungi, eg yeast, small, highly refractive bodies, known as microsomata, may be frequently observed. On applying pressure to the cover-glass placed on a pre- paration containing cells that exhibit such enclosures, the membranes areruptured and the mcrosomes are hberated — If, now, the latter be transferred to another8 THE THRORY Off SPONTAN . EOUS: GENERATION ' nutrient solution, there will most assurelly be develapment of organiams, if we omit those precautions which.are. considered: sssentjal by the bacteriologist, but superfluous by those who belisve in spantancous generation. Such development is, however, due, not to the mirogames, but to the germs introduced during the transfer, Although this 1s 60 evident, it 18 Strange thab this ew should have had its opponents, as, for instanog,, the botanists H. Karsten and A. ‘Wigand (Zand 11} and, with still greater pertinaeity, A. Béchamp. ‘The last-mentioned designates these microsomes (‘‘granulations moleculaires”) microzymes, -and sttributes to them such tenaclty gf. fe-that they are able to remain dormant, pot only for years, but even for enitine geological periods, since, as Béohamp asserts, ho has found mierozymes of cells which'were buried m the strata formed during fhe Oretaccous period still retaining their vitality and reproduchve power, A. fall account of this microzyme theory, which many amateur bactertologists have considered to be indisputable—commanications respecting which have been , > incessantly intruded upon the notice of the. Academy of Science at Paris—ia given in a bulky volume which Bicwamp (I) aid before his sceptical contemporaries am 1883, § 9.—Spontaneous Gengratien enly Unproven, not Impossible. Ons viowm eo ovo (every living éronture from an ogg); omnes vieum ec vivo monstrated ? Lot us devote a few moments to a onitical review of the question. Ono thing 1s established beyond doubt, namely, that all the instante of sup- Posed spontaneous generation brought forward by the adherents of the theory ave been vitisted by numerous errors It 1s, moreover, eatabiehed thet tho crourTence of spontancous generation has. not bees proved, n0/umagiailuble exper, ment beng known 1a whieh hyiz arenes eas rodvchd # fiom, inanimate substances. Spontaneous generation is therefore unproven, ae Whether also an ampossibtlity 1s a point still to be decided, if Sb: theory of evolution, as presented by Lamarck and Darwin, be traced towards ite origin in the lowest organisms, we come to a standstill wish the question “And from whenoe then comes the ultimate and lowest creature ?How did orgame hfe omginate on our globe ?” stn ToD fomnished by the Boglish phymcist Thomson that our earth was fertilised in its youth, inging the gi heavenly bodieg-spf ualinits i 3 F question: “How did life originate on’ these unknown, extra-mundane sources Of creative messengers?” There are only two poasible answers te thes questions, ‘viz., spontaneous generation, or a miracle «defn matter of reason, we aro therefore obliged to assume that, at some definite moment in the past, organised living beings were produced from un- * organised potentially organic substances, and farther, that such creative power may still be operating, may perhaps bo performing at present ‘The poserbality cannot be gansaid, ‘That bactena aro the result of this primary ereation of living beings is very aueshonable, and even improbable, sunce their structure 18 much ones compli. cated than is conastent with their presumed origin directly from chemical elements, unmodified by changes in" passing through smpler intermediate orgamems, 1 Lord Kelvin, s °UNPROVEN, NOT IMPOSSIBLE 9 Many investigators, and amongst them . Nigarz (L), assumed that these lowest forms really exist, although undiscovered at present, and in his important and highly suggestive work on the Theory of Descent—which also contains a fine chapter on “the limits of knowledge m natural science”—this author touches upon the question under consideration He calls these presumptive connecting links Probi8n (pre-exuting), on the ground of their being the pre- decessors of all known forms of living bemgs. Such a Probion resulting from spontaneous generation would be “merely a drop of homogeneous structureless Pltoma, devoid of any definite form and compoced of albaminates, uasociated only with the compounds necessary for nutrition.” “We must assume”—says de Bary—that organisms must at one time have originated from organisable but unorganised substances, without pro- nitors.. . . To prove such a primary creation of a hving beng is of the nghest interest, and exercises the same fascination on the investigator as the expectation of the homunculus in the phial did on the alchemst, Tho expe- rience of centuries has, however, shown that the homunculus when it actually appeared was simply a small imp which had been secretly passed into the fiasic by slaght of hand... . Therefore—admitting all 1maginable possibihties— the law, based on experience, of origin from ancestors, corresponds with the enlightened state of our knowledge, and this is tho starting-point thnt must be taken in a work which has to deal wath the exact sciences *qr THMORIES OF FERMENTATION. § 10.—Stahl’s Theory of Fermentation. ‘Waoxven was the first to leave the juice of sweet fruit to itself in storage for a fow days had the pleasure of observing a phenomenon hitherto unknown— the incipient decomposition of the mass—which we nowadays term alcoholic fermentation. This observation was made at so early a date that we have no record of it beyond myth and tradition -The Grocks féted the deity Bacchus as the inventor of wine, and the Egyptians ascribed to Osiris the first introduc- tuon of brewing. Acquaintance with the nature of this phenomenon was, however, of an extremely superficial character for a very long time Even in the later Middle ‘Ages the word fermeniaho (fermentation) was employed as synonymous with digestio (digestion), the latter word being also currently used to denote any form of chemical reaction ; and the word “ferment” was apphed to any body capable of producing such reaction. ‘At an early date xt would necessarily be noticed that the “must” when in a state of fermentation became covered with a froth, and that at the end of this operation a copious deposit, viz, yeast, was left at the bottom of the vessels Fermentation was therefore looked upon asa process of purification, by which the initially turbid and discoloured liquid was so mproved and freed from dirt, that the purified alcohol oxbited its true properties For this reason the deposit was described as the faces vini or feces cereniewt, ie, the excrement of the wine or beer. ‘This view was held by,eg Basilius Valentmus, a German monk and alchemist, who lived at Erfurt early m the fifteenth century It was also noticed that this sediment was a powerful ferment, 14,16 was capable of rapidly exciting a brisk fermentation an still unfermented liquids, such as wine-must or beer-wort. This idea was adopted in other branches of chemistry, so that any reaotion was considered as elucidated when the body acting as “ferment” there could be identified — Moreover, the “ philosopher's stone,” the goal of the labours and aspirations of the alchemists, was nothing but the much sought for, but never discovered, universal “ferment” for every possible chemical process! ‘Among the disciples of the alchemie school, one other, vz, Srant (I), deserves mention, because his views on the nature of fermentation were adopted by Liebig a hundred and forty years Jater. Stahl extended the defimtion of fermentation to all forms of decomposition, his theory bemg expressed verbatim as follows: “Patrefaction (and algo fermentation) is internal movement A body undergoing such internal movement may easily induce the same in any other body, which, though still quiescent, is susceptable of such movement.” § 11.—Gay-Lussac’s Opinion. Stabl’s view remained in vogue until the commencement of the mine- teenth century, when Gay-ussao, in 18r0, enunciated a new theory to a new 10GAY-LUSSAC'S OPINION Ir age. Tho discovery by Lavoumer that combustion 1s .a process of oxidation, a combination of oxygen with the combustible substance, was an event the influence whereof extended over the entire domain of chemistry, The assign- ment to oxygen of a part in the process of fermentation was therefore opportune ; but Gay-Litseac was especially prompted by another circumstance, ‘A Parisian confectioner and cook, named Appert, had made practical use of the experiment devised by hia contemporary Spallanzani for the refutation of the heterogenists and, after some prelimmary trials, perfected hns process for presorving meats, vegetables, spirituous hquors, &e.’ 'To this end be exposed them, im hermetically closed vessels, to the temperature of boilng water for some time—a process which had somewhat earher (r782) been recommended by the Swedish chemist Scxzmtz (I) for the conservation of vinegar. In this way Arpzar founded a new branch of industry—the manufacture of conserves— which brought hum both wealth and fame. He pubhshed a volume (I.) which comprised the results of his experience It was widely circulated and ran into severa] editions, the first of which appeared in 1810, and the fourth in 1831. It 18 therefore little matter for surprise that the attention of the Pansian chemist was directed (whether from the culinary or the literary side) to the productions of his enterprising fellow-atizen, Gay-Lvssac (1) now exammed conserves prepared according to Appert’s process, and found them to be free from gaseous oxygen. This incited him to make fermentation experiments ‘with wine-must, &¢, the results of which led him to assert that the presence of oxygen 1s necessary to the icapiion of fermentation. A number of over- zealous colleagues, in expounding their master’s opmion, added new features to xt, and subsequently credited him with the assertion that oxygen 1s the actual forment—a statement as unfounded as it is accurate, Gay-Lussac only claimed for the gas‘a eingle function, the inception of fermentation , once the process was, in operation the stimulus was no longer required. Wath regard to the nature of this stimulating action he was, however, unable to report more definitely. ‘Among the observations which led Glay-Lussac to adopt this view, mention may be made of one which appeared to him particularly conclusive, namely, the steriheation of wine-must by sulphuring. When wine-casks, before fillmg, are thoroughly sulphured—ie. the internal aur contained in them 1s heavily charged with sulphur dioxide by burning sulphur m the casks—the grape juice thereafter introduced remains quiet and passive, without fermenting. ‘This circumstance as now unanimously ascribed to the vitality of the yeast cells in the must beng destroyed by the sulphurous acid. Gay-Lussao, on the other hand, viewing the matter differently from his standpoint, held the opinion that as the sulphurous acid had a strong affimty for the oxygen, the two combined, and as no oxygen was available for starting the fermentation, the must necessarily remained inert. ‘The experiments made by Schwann in 1838, and described in § 13, refuted the opinion of Gay-Lussae, by demonstrating that the réle of exciting fermentation 1s set up by certain microscopic hving creatures which perform their functions am the absence of oxygen, Subsequent research proved that the presenco of this gns is altogether superfluous, so far as the progress of alcoholic fermentation 18 concerned, although it 18 not without influence thereon. Pasraur (II) mm 1861 established 1t as a fact that this progress 1s more satisfactorily effected when the fermenting quid 1s subjected to brisk aeration, § 12.—Cagniard-Latour's Vitalistic Theory of Fermentation. The French apothecary Astren (I. and IT ) has generally been credited with being the next mdividual, after Leeuwenhoek, who gave his attention to the nature of yeast. An examination of his published works shows, however, that12 THEORIES OF FERMENTATION hus investigations into fermentation were conducted without the aid of the microscope, so that he did not bring to hght any actual facts concerning the nature of yeast, but—as was ‘pointed out, though in vam, by Quavanws (I) a8 far back 3 1838—based his assumptions on hypotheses devoid of foundation, In the same way another Frenchman, viz., Desmaziéres, the reputed pioneer of the founders of the vitalistie theory of fermentation, cannot permanently retain this title, Laie Astior, he 1s said to havo recognised the part played by yeast in fermentation, but, as a reference to hus treatise, published in 1826 (in pages 42 to 67 of vol x. of the Ann, des Sc, Nat), will show, this assertion 18 incorrect. In these observations Desmazitres viewed the matter simply as a naturahet. ‘His investigations of the fungoid growths covering the surface of moist substrata were conducted from this point of view, and st was in the course of this study that he examined the mycelia that develop on beer, &ic. ‘These consist of masses of elongated cells, to which he gave the name Mycoderma cerensa, As ho fancied they exhibited powers of locomotion, he considered them as belonging to the animal Iangdom (anemaloula monadina), but, tne to his oroly dossnptive inchnations, he disregarded thew physiological properties, and especially the: influence on the substratum Thus the reputation attmbuted to Deamazidres of having, 12 1826, microscopically studied the morphology of the yeast-like cells, to which Person had defintely alluded four years earher, is dissipated by facts. On the other hand, a German worker, viz. Euxtepen (I), had already, mn 1838, correctly estimated the umportance of yeast, in that he asserted 1t to be a living organism, the vital functions of which are the cause of fermentation, Unfortunately ho did not follow up this 1dea, whuch was thrown out asa mere occasional remark m his treatise on practical analytical experiments, Otherwise he would, in 1818, have anticipated what was only accomplished twenty years Inter, viz., the establishment of the fact that (alcohoho) fermentation 1 causa- tively connected with the life (vita) of certain organisms, ‘This was determined, almost simultaneously, by three investigators working quite mdependently of each other Oagmurd-Latour i France, and Theodor Schwann and Friedrich Kutaing in Germany. The paths by which these three arnived at their common goal differed. ‘The versatile French techmiaist 1 known by namo to the majority of eduented people on account of the siren he invented, and which 1s largely used in the science of acoustics. He also devoted some attention to brewing, and compiled a work on the fermentation of beer. ‘The prelimmary studies undertaken 1n this connection Jed him to more closely investigate the nature of the “ yeast,” of which—notwith- standing the observations of his two compatriots already mentioned—practically nothing was thon known, ‘This material he exammed with the assistance of the microscope, and laid the resulta of his researches before the Parisian Academy on June 12, 1837, in a short peper (II,) contaming the following chief points 1 Beer-yeast, mstead of being an inammate chemeal substance, as previously supposed, actually consists of small globules which possess reproductive power, and are therefore living orgamsms. 2. Thess bodies appear to belong to the vegetable kingdom, and to reproduce themselves in two ways, 3+ They seem to act upon sugar solution only whilst still hving , wherefore 1t may, with great probability, be concluded that, by their vital activity, carbon dioxide 18 hberated, and the sugar solution transformed into an aleohohie quid § 18.—The Researches of Theodor Schwann. As the words printed in italies in the two preceding sentences show, and as 8 closer examination of the griginal treatise will more clearly reveal, Cagniard . ©THE RESEARCHES OF THEODOR SCHWANN 13 did not indubitably establish the vegetable nature of yeast. The accomphsh- ment of this task, and the attmbution of this organiem to its proper position in the system of Botany, formed the subject of a treatise published by Scawann (I.) am the first half of 1837, i.e contemporancously with Cagniard’s paper In following up the results of bis researches on spontaneous generation, Schwann studied beer-yeast, and found that the individual globules, of which the meas was seen under the microscope to consist, frequently became united into choin-hke or laterally branching bands, and presented to the eye an appearance greatly resembling that of many already well-known multicellular fungi. It was not this discovery alone, however, but rather thetr mode of reproduction, which induced Schwann to considér these bodies as of a vegetable nature In this process the globule pushes out from its interior 2 small nodule, which Schwann was able to observe develop to 1ts normal dimensions“ Observation of its growth leaves no doubt as to ite vegetable nature, since animals do not reproduce themselves in this manner.” ‘The rate of reproduction of the globules kept pace with the increasing briskness of the fermentation, so that Schwann came to the opmion that 1t was highly probable that the development of the fermentation was induced by that of the organum “Vinous fermentation must therefore be regarded as the decomposition occasioned by the sugar fungus extracting, fiom the sugar and a nitrogenous body, the materials necessary toitsnutrition and giowth, whereby such elements of these bodies (probably among other substances) as are not taken up by the plant umte, by preference, to form alcohol” Thus discovery was communicated by Schwann to his friend and colleague, Professor Meyen, who tested and confirmed it, “stating with reference thereto, that the only doubt arising was whether the organism in question was an alga or a thread fungus, the latter seeming the move likely by reason of the absence of green pigment.” Thus yeast was recogmued as a fungus, and, fiom its capacity of fermenting sugar, was designated sugar fungus whence the current generic name, Saccharomyoes Meyen. Aocording as such a sugar fungus was found active 1n beer-wort or wine-must, x6 was called by the specific name of S cereounw or S. vit, which names remained in general use in ther original sigmficance until ReEs (I) in 1870 proposed a system of differentiation which will be more fully noticed in a subsequent aragruph, Patifs fellows from the remarks already made, the name “ yeast” apphed merely to one particular group of ferments, viz., those producing alcoholic fermentation. For a considerable period after Cagmiard’s discovery, however, 1 was used indis- eruminately for all ferments ‘Thus, for example, Pasteur speaks of the “ yeast” of lactic fermentation, meaning thereby Bacteria , and even m 1879 Nagel, the investigator of the fission fungi, refers in his “Theory of Fermentation” to the “yeast” of putrescent urme ‘This misuse of the term has been abandoned, and the name “ yeast” 1s now only employed when speaking of the budding fungi that excite alcoholic fermentation. § 14.—Friedrich Kiitzing’s General Theory of Fermentation. Tho views promulgated by this German worker m the field of Vegetable Physiology and the Algw were in harmony with the spirit manifested in the “ Elements of Philosophie Botany ” Published almost stmultaneously with the above-mentioned communications of Ongnimd-Latour and Th. Schwann—though actually compiled at a much earher date (before 1834)—Kutzng’s treatise (I) on this subject surpassed those of his two colleagues in more than one particular ‘The value of his14 ‘THEORIES OF FERMENTATION actual determmations 1s not, in our opinion, lessened by the fact that he was an. advocate for spontaneous generation, since at that time (in 1837) there exuted no decisive and unassailable proofs to controvert this theory Kutamg did not restrict his researches solely to alcoholic fermentation, but also instituted comparisons with number of other similar phenomena, regard- ing them all from the same point of view. Even though he must share with others the credit of having discovered the organised structure of yeast, that of determining the vegotable nature of the “mother of vinegar” and recognising its mode of action belongs to him alone With these discoveries are associated a number of others of minor importance, such, for example, as the physiological ‘basis of the method (propounded by Scheele) of preparing gallic acid by allowing fa solution of pyrogallic and (eg. gall-nut extract) to become mfested with mould. The numerous phenomena he brings under our notice constitute so many puoofs of the theory that fermentation cannot be regarded as a purely chemal process ‘‘ It 1s well known that chemistry explains vinous fermenta- tion by the reaction of the so-called gluten on the amylum (starch) and sugar. I must firmly maintain that the explanation does not give me a clear idea of the process, and Iam inclined to doubt whether others are more fortunate in this respect It 18, however, certain that the entire process of alcoholic fermen- tation 18 dependent on the formation of yeast, and the acid fermentation on the formation of the vinegar plant, . . . long with the inerassed growth of these organsma the reproductive mpulse also increases, and,' concurrently, their rouction on the hquid present. . . . In so far aa fermentation 1s synouymous with a reciprocal reaction of organic and inorgame bodies on the constituents of a given liquid which may be regarded as forming the nutrient medium of the organic product, so 15 it necessarily synonymous with overy organio vital fune- tion wherefore organio hfe = fermentation. On the other hand, such pro- ceases as lead to the production of vinegar from alcohol by the use of platinum black or other similar methods, cannot be compered with fermentation, beng purely chemical, whilst fermentation 1s an organo-chemical process, as 18 also the hfe process of auy organic body ” ~ One of the three members of the committee appointed by the Académie des Sciences of Paris to report on the memoir presented by Cagniard—namely, Tonpw (I.)—took the opportunity thus afforded of expenmentally dilating upon his compatriot’s work, and of amalgamating these new “ discoveries ” with the revelations of Schwann and Kntang. In ths way a volume, containing more pages than Latour’s communication had columns, came ito existence, without, however, adding to our knowledge in the shghtest degree. Turpin seems, however, to have thoroughly known his public, since he 1s even now regarded as one of the founders of the vitalistic theory of fermentation, not only by compilers of text-books, but also by actual investigators, from whom. one might more reasonably expect @ more thorough study of the original works of their predecessors, 7 § 15.—Liebig’s Decomposition Theory. ‘Two years subsequent to the pubheation of the works of Oagniard, Kutzing, and Schwann, Inebig placed before his colleagues a now theory, aocording to which fermentation was a purely chemical reaction. In order to avd judging this chemist unreasonably, one must bear in mind the ago wherein thia theory was promulgated Synthetic orgame chemistry had just been founded —Eloven years previously (1828) Wobler had succeeded im artificially preparing urea, to the astonishment of his contemporaries, who had hitherto considered ag umpossible the artificial production of organic‘ i 15 substances outeld . ble body of whose vital functions they are the outcome, 5 could not be produced without the concurrence of i. an established mamm. ‘To overthrow this dog #ies that any desired organic substance can be prepared without thy Sawummsee sc? vital action, was the endeavour of the majority of the cher Tnebig being one of the foremost, most industrious, and st én the cause. It 18, therefore, small matter for astonsat Beveguard-Latour, Kutang, and Schwann to the contrary, notwi could concave a theory of fermentation wherem the action 6 ms had no place and their vital force was ignored. ‘The struggle t considered to be the objectionable theory of these three phystdlo ned by Liebig in 1839 with an anonymous treatise (I.), im whielf" srvations of the microscopists we1e covered with highly amusing # the next year he returned to the charge in earnest in his werkt & fils Ohomistry in Relation to Agriculture and Physiology,” on pp* bivRich his new theory 1s enunciated. This is, as already mentioned; frase ‘put forward by Stahl more than a century before : Laebig considet on as molecular movement, which a body in a state of chemical maiieines eoomposition, transfers to other substances whose clements ato ly combined. Between fermentation (m its lumited sense) and ff : v@ is the following difference : In the latter— putrefaction—the ‘trenamitted by the decomposing material— wiz,, the albumne! BMiefaction, once begun, is continued by inherent movement, even’ cause has been rendered mactive With fermentation it is offre “it this process the body (sugar) m a state of incipient decomposition cainét tranemit the movement to the still undecom- posed substance Consequently this function lias to be performed by an extra- neous causative agent, a ferment, which in this cage 18 necessary not only for commencing (ss with putrefaction), but also for continuing the decomposition. Temast-be admitted thet, ab fret aight, this definition, as alsothe differen tia- tion between fermentation and putrefaction, 1s very attractive. Nevertheless xt will not bear the light of Keen eriticum Take, in the first place, the character on which the distinction between fermentation and putrefaction 1s based, viz , that the former will not go on without the presence of the ferment, whereas, on the other hand, putrefaction, when once started, continues spontaneously, the ferment being no longer needed. The reason why Liebig was induced to make ths distmetion 1 easy to futhom, In the case of fermenting beor- wort—which Inebg usually had in view when speaking of fermentation —the forment (beer-yesst) was discernible to the naked eye, and experience tought that without this ferment the fermentation could not be satisfactorily carnied on. On the other hand, the presence of those mmute organisms which, as we now know, insinuate themselves into all substances liable to putrefaction, and decompose the same without, as a rule, giving rise to such a multipheation of the deposited ferment as can be remarked by the mexpert eye, 18 nob go aummediately apparent as in alecholit fermentation. ‘Thus, even an Lisbig’s opinion, yeast is etrantial to, the eontumaance of fermentation, only, the ferment is degraded to a simple albummord substance. To enter nowadays ito a further onslaught agaist this theory would be merely storming an undefended position, Moreover, as the subsequent editions of the aforesaid work demonstrate, xt was gradually modified by its author, so that the form im which 1 was presented in his latest exposition (II.) in 1870 differs in many particularé from the original. °16 THEORIES OF FERMENTATION The ocular demonstration, in individual instances, of the untenable nature of the hypotheses supporting this theory, to those whom the representations of Latour, Kutzing, and Schwann had not suoceeded in convincing, was the congenial ‘task undertaken by Pasteur, and brought by hm to a successful issue with great experimental skill. § 16,—Pasteur’s Theory of Fermentation (III.). ‘The victorious autagonist of the theory of spontaneous generation was not content with controverting the views of Liebig, he also sought to erect a better theory in ite stead. According to this doctrine, 1t 15 the Jack of free oxygen that eads to the fermentation being set up by the orgamsm as a means of supplying ateolf with the energy it requires by eerzing upon the oxygen thereby obtain able, “Formontation 1s life without air” A very slight experience in this matter suffices for the recognition of the fact that this theory takes no account of tho several kinds of fermentation in which the presence of oxygen is a neces- sary condition, viz , the so-called oxidation fermentations—the best example of which 1s afforded by the scetuc fermentation, In this respect the theory cannot be further alluded to in the present chapter, which 1s devoted to general con- siderations, It will be fully dealt with 1 a subsequent chapter. ‘The only remark to be made now is that this theory also has proved untenable ‘The permanent value of the services rendered to Fermentation Phymology by Pasteur are not diminished by the disproval of his theory of fermentation, since they have their root in the successful endeavour, by means of careful and oxtensive experimental demonstration, to bring nto universal recognition the theory originated—but only imperfectly formulated—by Oagniard, Kutzing, and Schwann, of the causative connection between fermentation and the vital activity of the microbe; § 17,—C. Nigeli’s Physico-Molecular Theory. Although it was by this timo indubitably estabbshed that without the vital activity of micro-orgamsms no fermentation could occur, no clear account bad been given as to how the achvity itself was exerted. ' Several explanations were possible According to one which was especially advocated by Kutzing and Pasteur, a decomposition was effected within the cells of the organic ferments, which obtained ther nourishment from the fermenting material (e.g. sugar) and discharged the fermentation products as waste matter, According to another, the decomposing force suaply emanated from the cells and became tho direct cause of the decomponition of the fermentable matter around them, The physico-molecular theory, proposed by Niaurt (Il), expresses this view in the following words “Fermentation is, therefore, the transference of con- ditions of movement in the molecules, atomic groups, and atoms of the various compounds constituting the hving plasma (which compounds remain chemically uachanged) to the fermentative material, whereby the equilibrium of 1ts molecules 1s destroyed and their dissociation induced” ‘The radzus of the sphere of influence of the individual yeast cells is estimated by Nageli as from 20 to 50 p. ‘This definition differs from that formerly given by Liebig merely in a single, though important, consideration, viz, 1t regards the hving cell as the source of action, whereas the other definition speaks of manmmate albuminoid substances. ‘Nageli was, however, unable to prove the correctness of his theory,and the calcula~ tions deduced from other observations which he brought forward in support thereof have in course of time proved inapplicable,3 THE ENZYMES AND M. TRAUBE’S THEORY 7 § 18.—The Enzymes and M. Traube’s Ferment Theory. ‘There is still another possible explavation of the power of living cells to act ata distance, This regards the cells, not as centres of radiating molecular movements, but as forming centres of production of metabolic products which penetrate through the cell membrane into the surrounding hqud. There they become widely distributed by diffusion, and by their influence bring about the decomposition of certain constituents of the solution, but do not undergo any chemical change themselves. These active bodies are called enzymes, ther behaviour is, as will be observed, different from that of ordinary chemical agents, since the latter effect alterations in other groups of atoms by ther chemucal affimty, whereby the old combination 1s broken up, and tho separated portion enters into a new atomic grouping with a part of the activeagent. Accordingly, a dofimte weight of the agent can only displace a definite quantity (known as the “equivalent weight”) of other compounds; whereas the enzymes behave differently, thetr activity being pracheally wimitable. They do not combine with the pro- ducts of the reaction, but continue to act on the residual undecomposed substance. ‘The first enzyme was discovered by Paysn and Persoz (I.) in 1833, who detected in malt extract a substance—which they termed diastase—capable of converting starch into sugar. ‘They were, however, unable to isolate it mn a pure condition, Threo years later Tropor Scuwany (IIT.) discovered in gastric juice pepsin, subsequently also nated peptase, which in faintly acid solutions resolved undiffusible albumen into assimilable dissociation products. Since that time the same enzyme has also been detected m varions vegetable organisms, many vaneties of bacteria in particular having the power of elaborating 1t. There will be ample opportunity for reference to this pont along with the other known enzymesat e subsequent atage. At present we have only to consider them aa the basis of « theory of fermentation, the formulation of which dates back as far es 1858, but has come to the front more of late years, and—so far as can be judged from the data at present available—will acquire still greater importance. ‘As we have observed in § 10, the meaning attached, under the influence of alchemical views, to the word ferment was, until the close of the eighteenth century, very comprehensive, and 1t was only then that the restriction of the term to bodies inciting fermentation began. Contemporaneous with the develop- ment of positive knowledge with regard to these bodies was the discovery of the enzymes, the behaviour of which resembled that of the former, in so far that they exhibited a capacity of mducing decompoation. Moreover, the obscunty in which these organisms were still enshrouded was equally mysterious in both cases, and, since the organic ature of the true instigators of fermentation was eather unknown or was not considered worth attention by the chemists of the day, at happened that the name “ferment” was also applied to the newly discovered enzymes With an increasing insight into the true state of the matter grew the conviction that two very different things had been grouped under one name, and this conviction found expression in the distinction thenceforward of the true inatigators of fermentation as organised or structural ferments, whilst the enzymes were designated unorganised or structureless ferments. ‘These terms are till current 10 chemical text-books, whereasin Fermentation Physiologyit 1s customary to speak merely of fermentative organisms on the one hand and of enzymes on the other. ‘M, Taavae (I. and IL) in 1858 made the origin and mfluence of these enzymes the basis of a new conception (Ferment Theory) of fermentation, according to which this process is not instigated by the organisms themselves, but by the enzymes formed es products of their vitality and excreted by them, 1 . B