THE STORY OF GERM LIFE

BY H. W. CONN

PROFESSOR OF BIOLOGY AT WESLEYAN UNIVERSITY,

AUTHOR OF EVOLUTION OF TO-DAY, THE LIVING WORLD, ETC.



PREFACE.

Since the first edition of this book was published the popular idea of bacteria to which attention was drawn in the original preface has undergone considerable modification. Experimental medicine has added constantly to the list of diseases caused by bacterial organisms, and the general public has been educated to an adequate conception of the importance of the germ as the chief agency in the transmission of disease, with corresponding advantage to the efficiency of personal and public hygiene. At the same time knowledge of the benign bacteria and the enormous role they play in the industries and the arts has become much more widely diffused. Bacteriology is being studied in colleges as one of the cultural sciences; it is being widely adopted as a subject of instruction in high schools; and schools of agriculture and household science turn out each year thousands of graduates familiar with the functions of bacteria in daily life. Through these agencies the popular misconception of the nature of micro- organisms and their relations to man is being gradually displaced by a general appreciation of their manifold services. It is not unreasonable to hope that the many thousands of copies of this little manual which have been circulated and read have contributed materially to that end. If its popularity is a safe criterion, the book has amply fulfilled its purpose of placing before the general reader in a simple and direct style the main facts of bacteriology. Beginning with a discussion of the nature of bacteria, it shows their position in the scale of plant and animal life. The middle chapters describe the functions of bacteria in the arts, in the dairy, and in agriculture. The final chapters discuss the relation of bacteria to disease and the methods by which the new and growing science of preventive medicine combats and counteracts their dangerous powers.

JULY, 1915.



CONTENTS.

I.—BACTERIA AS PLANTS

Historical.—Form of bacteria.—Multiplication of bacteria.—Spore formation.—Motion.—Internal structure.—Animals or plants?— Classification.—Variation.—Where bacteria are found.

II.—MISCELLANEOUS USES OF BACTERIA IN THE ARTS.

Maceration industries.—Linen.—Jute.—Hemp.—Sponges.—Leather. —Fermentative industries.—Vinegar—Lactic acid.—Butyric acid.— Bacteria in tobacco curing.—Troublesome fermentations.

III.—BACTERIA IN THE DAIRY.

Sources of bacteria in milk.—Effect of bacteria on milk.— Bacteria used in butter making.—Bacteria in cheese making.

IV.—BACTERIA IN NATURAL PROCESSES.

Bacteria as scavengers.—Bacteria as agents in Nature's food cycle.—Relation of bacteria to agriculture.—Sprouting of seeds. —The silo.—The fertility of the soil.—Bacteria as sources of trouble to the farmer.—Coal formation.

V.—PARASITIC BACTERIA AND THEIR RELATION TO DISEASE

Method of producing disease.—Pathogenic germs not strictly parasitic.—Pathogenic germs that are true parasites.—What diseases are due to bacteria.—Variability of pathogenic powers.— Susceptibility of the individual.—Recovery from bacteriological diseases.—Diseases caused by organisms other than bacteria.

VI.—METHODS OF COMBATING PARASITIC BACTERIA

Preventive medicine.—Bacteria in surgery.—Prevention by inoculation.—Limits of preventive medicine.—Curative medicine. —Drugs—Vis medicatrix naturae.—Antitoxines and their use.— Conclusion.



THE STORY OF GERM LIFE.

CHAPTER I.

BACTERIA AS PLANTS.

During the last fifteen years the subject of bacteriology [Footnote: The term microbe is simply a word which has been coined to include all of the microscopic plants commonly included under the terms bacteria and yeasts.] has developed with a marvellous rapidity. At the beginning of the ninth decade of the century bacteria were scarcely heard of outside of scientific circles, and very little was known about them even among scientists. Today they are almost household words, and everyone who reads is beginning to recognise that they have important relations to his everyday life. The organisms called bacteria comprise simply a small class of low plants, but this small group has proved to be of such vast importance in its relation to the world in general that its study has little by little crystallized into a science by itself. It is a somewhat anomalous fact that a special branch of science, interesting such a large number of people, should be developed around a small group of low plants. The importance of bacteriology is not due to any importance bacteria have as plants or as members of the vegetable kingdom, but solely to their powers of producing profound changes in Nature. There is no one family of plants that begins to compare with them in importance. It is the object of this work to point out briefly how much both of good and ill we owe to the life and growth of these microscopic organisms. As we have learned more and more of them during the last fifty years, it has become more and more evident that this one little class of microscopic plants fills a place in Nature's processes which in some respects balances that filled by the whole of the green plants. Minute as they are, their importance can hardly be overrated, for upon their activities is founded the continued life of the animal and vegetable kingdom. For good and for ill they are agents of neverceasing and almost unlimited powers.

HISTORICAL.

The study of bacteria practically began with the use of the microscope. It was toward the close of the seventeenth century that the Dutch microscopist, Leeuwenhoek, working with his simple lenses, first saw the organisms which we now know under this name, with sufficient clearness to describe them. Beyond mentioning their existence, however, his observations told little or nothing. Nor can much more be said of the studies which followed during the next one hundred and fifty years. During this long period many a microscope was turned to the observation of these minute organisms, but the majority of observers were contented with simply seeing them, marvelling at their minuteness, and uttering many exclamations of astonishment at the wonders of Nature. A few men of more strictly scientific natures paid some attention to these little organisms. Among them we should perhaps mention Von Gleichen, Muller, Spallanzani, and Needham. Each of these, as well as others, made some contributions to our knowledge of microscopical life, and among other organisms studied those which we now call bacteria. Speculations were even made at these early dates of the possible causal connection of these organisms with diseases, and for a little the medical profession was interested in the suggestion. It was impossible then, however, to obtain any evidence for the truth of this speculation, and it was abandoned as unfounded, and even forgotten completely, until revived again about the middle of the 19th century. During this century of wonder a sufficiency of exactness was, however, introduced into the study of microscopic organisms to call for the use of names, and we find Muller using the names of Monas, Proteus, Vibrio, Bacillus, and Spirillum, names which still continue in use, although commonly with a different significance from that given them by Muller. Muller did indeed make a study sufficient to recognise the several distinct types, and attempted to classsify these bodies. They were not regarded as of much importance, but simply as the most minute organisms known.

Nothing of importance came from this work, however, partly because of the inadequacy of the microscopes of the day, and partly because of a failure to understand the real problems at issue. When we remember the minuteness of the bacteria, the impossibility of studying any one of them for more than a few moments at a time —only so long, in fact, as it can be followed under a microscope; when we remember, too, the imperfection of the compound microscopes which made high powers practical impossibilities; and, above all, when we appreciate the looseness of the ideas which pervaded all scientists as to the necessity of accurate observation in distinction from inference, it is not strange that the last century gave us no knowledge of bacteria beyond the mere fact of the existence of some extremely minute organisms in different decaying materials. Nor did the 19th century add much to this until toward its middle. It is true that the microscope was vastly improved early in the century, and since this improvement served as a decided stimulus to the study of microscopic life, among other organisms studied, bacteria received some attention. Ehrenberg, Dujardin, Fuchs, Perty, and others left the impress of their work upon bacteriology even before the middle of the century. It is true that Schwann shrewdly drew conclusions as to the relation of microscopic organisms to various processes of fermentation and decay—conclusions which, although not accepted at the time, have subsequently proved to be correct. It is true that Fuchs made a careful study of the infection of "blue milk," reaching the correct conclusion that the infection was caused by a microscopic organism which he discovered and carefully studied. It is true that Henle made a general theory as to the relation of such organisms to diseases, and pointed out the logically necessary steps in a demonstration of the causal connection between any organism and a disease. It is true also that a general theory of the production of ail kinds of fermentation by living organisms had been advanced. But all these suggestions made little impression. On the one hand, bacteria were not recognised as a class of organisms by themselves—were not, indeed, distinguished from yeasts or other minute animalcuise. Their variety was not mistrusted and their significance not conceived. As microscopic organisms, there were no reasons for considering them of any more importance than any other small animals or plants, and their extreme minuteness and simplicity made them of little interest to the microscopist. On the other hand, their causal connection with fermentative and putrefactive processes was entirely obscured by the overshadowing weight of the chemist Liebig, who believed that fermentations and putrefactions were simply chemical processes. Liebig insisted that all albuminoid bodies were in a state of chemically unstable equilibrium, and if left to themselves would fall to pieces without any need of the action of microscopic organisms. The force of Liebig's authority and the brilliancy of his expositions led to the wide acceptance of his views and the temporary obscurity of the relation of microscopic organisms to fermentative and putrefactive processes. The objections to Liebig's views were hardly noticed, and the force of the experiments of Schwann was silently ignored. Until the sixth decade of the century, therefore, these organisms, which have since become the basis of a new branch of science, had hardly emerged from obscurity. A few microscopists recognised their existence, just as they did any other group of small animals or plants, but even yet they failed to look upon them as forming a distinct group. A growing number of observations was accumulating, pointing toward a probable causal connection between fermentative and putrefactive processes and the growth of microscopic organisms; but these observations were known only to a few, and were ignored by the majority of scientists.

It was Louis Pasteur who brought bacteria to the front, and it was by his labours that these organisms were rescued from the obscurity of scientific publications and made objects of general and crowning interest. It was Pasteur who first successfully combated the chemical theory of fermentation by showing that albuminous matter had no inherent tendency to decomposition. It was Pasteur who first clearly demonstrated that these little bodies, like all larger animals and plants, come into existence only by ordinary methods of reproduction, and not by any spontaneous generation, as had been earlier claimed. It was Pasteur who first proved that such a common phenomenon as. the souring of milk was produced by microscopic organisms growing in the milk. It was Pasteur who first succeeded in demonstrating that certain species of microscopic organisms are the cause of certain diseases, and in suggesting successful methods of avoiding them. All these discoveries were made in rapid succession. Within ten years of the time that his name began to be heard in this connection by scientists, the subject had advanced so rapidly that it had become evident that here was a new subject of importance to the scientific world, if not to the public at large. The other important discoveries which Pasteur made it is not our purpose to mention here. His claim to be considered the founder of bacteriology will be recognised from what has already been mentioned. It was not that he first discovered the organisms, or first studied them; it was not that he first suggested their causal connection with fermentation and disease, but it was because he for the first time placed the subject upon a firm foundation by proving with rigid experiment some of the suggestions made by others, and in this way turned the attention of science to the study of micro-organisms.

After the importance of the subject had been demonstrated by Pasteur, others turned their attention in the same direction, either for the purpose of verification or refutation of Pasteur's views. The advance was not very rapid, however, since bacteriological experimentation proved to be a subject of extraordinary difficulty. Bacteria were not even yet recognised as a group of organisms distinct enough to be grouped by themselves, but were even by Pasteur at first confounded with yeasts. As a distinct group of organisms they were first distinguished by Hoffman in 1869, since which date the term bacteria, as applying to this special group of organisms, has been coming more and more into use. So difficult were the investigations, that for years there were hardly any investigators besides Pasteur who could successfully handle the subject and reach conclusions which could stand the test of time. For the next thirty years, although investigators and investigations continued to increase, we can find little besides dispute and confusion along this line. The difficulty of obtaining for experiment any one kind of bacteria by itself, unmixed with others (pure cultures), rendered advance almost impossible. So conflicting were the results that the whole subject soon came into almost hopeless confusion, and very few steps were taken upon any sure basis. So difficult were the methods, so contradictory and confusing the results, because of impure cultures, that a student of to-day who wishes to look up the previous discoveries in almost any line of bacteriology need hardly go back of 1880, since he can almost rest assured that anything done earlier than that was more likely to be erroneous than correct.

The last fifteen years have, however, seen a wonderful change. The difficulties had been mostly those of methods of work, and with the ninth decade of the century these methods were simplified by Robert Koch. This simplification of method for the first time placed this line of investigation within the reach of scientists who did not have the genius of Pasteur. It was now possible to get pure cultures easily, and to obtain with such pure cultures results which were uniform and simple. It was now possible to take steps which had the stamp of accuracy upon them, and which further experiment did not disprove. From the time when these methods were thus made manageable the study of bacteria increased with a rapidity which has been fairly startling, and the information which has accumulated is almost formidable. The very rapidity with which the investigations have progressed has brought considerable confusion, from the fact that the new discoveries have not had time to be properly assimilated into knowledge. Today many facts are known whose significance is still uncertain, and a clear logical discussion of the facts of modern bacteriology is not possible. But sufficient knowledge has been accumulated and digested to show us at least the direction along which bacteriological advance is tending, and it is to the pointing out of these directions that the following pages will be devoted.

WHAT ARE BACTERIA?

The most interesting facts connected with the subject of bacteriology concern the powers and influence in Nature possessed by the bacteria. The morphological side of the subject is interesting enough to the scientist, but to him alone. Still, it is impossible to attempt to study the powers of bacteria without knowing something of the organisms themselves. To understand how they come to play an important part in Nature's processes, we must know first how they look and where they are found. A short consideration of certain morphological facts will therefore be necessary at the start.

FORM OF BACTERIA.

In shape bacteria are the simplest conceivable structures. Although there are hundreds of different species, they have only three general forms, which have been aptly compared to billiard balls, lead pencils, and corkscrews. Spheres, rods, and spirals represent all shapes. The spheres may be large or small, and may group themselves in various ways; the rods may be long or short, thick or slender; the spirals may be loosely or tightly coiled, and may have only one or two or may have many coils, and they may be flexible or stiff; but still rods, spheres, and spirals comprise all types.

In size there is some variation, though not very great. All are extremely minute, and never visible to the naked eye. The spheres vary from 0.25 u to 1.5 u (0.000012 to 0.00006 inches). The rods may be no more than 0.3 u in diameter, or may be as wide as 1.5 u to 2.5 u, and in length vary all the way from a length scarcely longer than their diameter to long threads. About the same may be said of the spiral forms. They are decidedly the smallest living organisms which our microscopes have revealed.

In their method of growth we find one of the most characteristic features. They universally have the power of multiplication by simple division or fission. Each individual elongates and then divides in the middle into two similar halves, each of which then repeats the process. This method of multiplication by simple division is the distinguishing mark which separates the bacteria from the yeasts, the latter plants multiplying by a process known as budding. Fig. 2 shows these two methods of multiplication.

While all bacteria thus multiply by division, certain differences in the details produce rather striking differences in the results. Considering first the spherical forms, we find that some species divide, as described, into two, which separate at once, and each of which in turn divides in the opposite direction, called Micrococcus, (Fig. 3). Other species divide only in one direction. Frequently they do not separate after dividing, but remain attached. Each, however, again elongates and divides again, but all still remain attached. There are thus formed long chains of spheres like strings of beads, called Streptococci (Fig. 4). Other species divide first in one direction, then at right angles to the first division, and a third division follows at right angles to the plane of the first two, thus producing solid groups of fours, eights, or sixteens (Fig 5), called Sarcina. Each different species of bacteria is uniform in its method of division, and these differences are therefore indications of differences in species, or, according to our present method of classification, the different methods of division represent different genera. All bacteria producing Streptococcus chains form a single genus Streptococcus, and all which divide in three division planes form another genus, Sarcina, etc.

The rod-shaped bacteria also differ somewhat, but to a less extent. They almost always divide in a plane at right angles to their longest dimension. But here again we find some species separating immediately after division, and thus always appearing as short rods (Fig. 6), while others remain attached after division and form long chains. Sometimes they appear to continue to increase in length without showing any signs of division, and in this way long threads are formed (Fig. 7). These threads are, however, potentially at least, long chains of short rods, and under proper conditions they will break up into such short rods, as shown in Fig. 7a. Occasionally a rod species may divide lengthwise, but this is rare. Exactly the same may be said of the spiral forms. Here, too, we find short rods and long chains, or long spiral filaments in which can be seen no division into shorter elements, but which, under certain conditions, break up into short sections.

RAPIDITY OF MULTIPLICATION.

It is this power of multiplication by division that makes bacteria agents of such significance. Their minute size would make them harmless enough if it were not for an extraordinary power of multiplication. This power of growth and division is almost incredible. Some of the species which have been carefully watched under the microscope have been found under favourable conditions to grow so rapidly as to divide every half hour, or even less. The number of offspring that would result in the course of twenty-four hours at this rate is of course easily computed. In one day each bacterium would produce over 16,500,000 descendants, and in two days about 281,500,000,000. It has been further calculated that these 281,500,000,000 would form about a solid pint of bacteria and weigh about a pound. At the end of the third day the total descendants would amount to 47,000,000,000,000, and would weigh about 16,000,000 pounds. Of course these numbers have no significance, for they are never actual or even possible numbers. Long before the offspring reach even into the millions their rate of multiplication is checked either by lack of food or by the accumulation of their own excreted products, which are injurious to them. But the figures do have interest since they show faintly what an unlimited power of multiplication these organisms have, and thus show us that in dealing with bacteria we are dealing with forces of almost infinite extent.

This wonderful power of growth is chiefly due to the fact that bacteria feed upon food which is highly organized and already in condition for absorption. Most plants must manufacture their own foods out of simpler substances, like carbonic dioxide (Co2) and water, but bacteria, as a rule, feed upon complex organic material already prepared by the previous life of plants or animals. For this reason they can grow faster than other plants. Not being obliged to make their own foods like most plants, nor to search for it like animals, but living in its midst, their rapidity of growth and multiplication is limited only by their power to seize and assimilate this food. As they grow in such masses of food, they cause certain chemical changes to take place in it, changes doubtless directly connected with their use of the material as food. Recognising that they do cause chemical changes in food material, and remembering this marvellous power of growth, we are prepared to believe them capable of producing changes wherever they get a foothold and begin to grow. Their power of feeding upon complex organic food and producing chemical changes therein, together with their marvellous power of assimilating this material as food, make them agents in Nature of extreme importance.

DIFFERENCES BETWEEN DIFFERENT SPECIES OF BACTERIA.

While bacteria are thus very simple in form, there are a few other slight variations in detail which assist in distinguishing them. The rods are sometimes very blunt at the ends, almost as if cut square across, while in other species they are more rounded and occasionally slightly tapering. Sometimes they are surrounded by a thin layer of some gelatinous substance, which forms what is called a capsule (Fig. 10). This capsule may connect them and serve as a cement, to prevent the separate elements of a chain from falling apart.

Sometimes such a gelatinous secretion will unite great masses of bacteria into clusters, which may float on the surface of the liquid in which they grow or may sink to the bottom. Such masses are called zoogloea, and their general appearance serves as one of the characters for distinguishing different species of bacteria (Fig. 10, a and b). When growing in solid media, such as a nutritious liquid made stiff with gelatine, the different species have different methods of spreading from their central point of origin. A single bacterium in the midst of such a stiffened mass will feed upon it and produce descendants rapidly; but these descendants, not being able to move through the gelatine, will remain clustered together in a mass, which the bacteriologist calls a colony. But their method of clustering, due to different methods of growth, is by no means always alike, and these colonies show great differences in general appearance. The differences appear to be constant, however, for the same species of bacteria, and hence the shape and appearance of the colony enable bacteriologists to discern different species (Fig. II). All these points of difference are of practical use to the bacteriologist in distinguishing species.

SPORE FORMATION.

In addition to their power of reproduction by simple division, many species of bacteria have a second method by means of spores. Spores are special rounded or oval bits of bacteria protoplasm capable of resisting adverse conditions which would destroy the ordinary bacteria. They arise among bacteria in two different methods.

Endogenous spores.—These spores arise inside of the rods or the spiral forms (Fig. 12). They first appear as slight granular masses, or as dark points which become gradually distinct from the rest of the rod. Eventually there is thus formed inside the rod a clear, highly refractive, spherical or oval spore, which may even be of a greater diameter than the rod producing it, thus causing it to swell out and become spindle formed [Fig. 12 c]. These spores may form in the middle or at the ends of the rods (Fig. 12). They may use up all the protoplasm of the rod in their formation, or they may use only a small part of it, the rod which forms them continuing its activities in spite of the formation of the spores within it. They are always clear and highly refractive from containing little water, and they do not so readily absorb staining material as the ordinary rods. They appear to be covered with a layer of some substance which resists the stain, and which also enables them to resist various external agencies. This protective covering, together with their small amount of water, enables them to resist almost any amount of drying, a high degree of heat, and many other adverse conditions. Commonly the spores break out of the rod, and the rod producing them dies, although sometimes the rod may continue its activity even after the spores have been produced.

Arthrogenous spores (?).—Certain species of bacteria do not produce spores as just described, but may give rise to bodies that are sometimes called arthrospores. These bodies are formed as short segments of rods. A long rod may sometimes break up into several short rounded elements, which are clear and appear to have a somewhat increased power of resisting adverse conditions. The same may happen among the spherical forms, which only in rare instances form endogenous spores. Among the spheres which form a chain of streptococci some may occasionally be slightly different from the rest. They are a little larger, and have been thought to have an increased resisting power like that of true spores (Fig. 13 b). It is quite doubtful, however, whether it is proper to regard these bodies as spores. There is no good evidence that they have any special resisting power to heat like endogenous spores, and bacteriologists in general are inclined to regard them simply as resting cells. The term arthrospores has been given to them to indicate that they are formed as joints or segments, and this term may be a convenient one to retain although the bodies in question are not true spores.

Still a different method of spore formation occurs in a few peculiar bacteria. In this case (Fig. 14) the protoplasm in the large thread breaks into many minute spherical bodies, which finally find exit. The spores thus formed may not be all alike, differences in size being noticed. This method of spore formation occurs only in a few special forms of bacteria.

The matter of spore formation serves as one of the points for distinguishing species. Some species do not form spores, at least under any of the conditions in which they have been studied. Others form them readily in almost any condition, and others again only under special conditions which are adverse to their life. The method of spore formation is always uniform for any single species. Whatever be the method of the formation of the spore, its purpose in the life of the bacterium is always the same. It serves as a means of keeping the species alive under conditions of adversity. Its power of resisting heat or drying enables it to live where the ordinary active forms would be speedily killed. Some of these spores are capable of resisting a heat of 180 degrees C. (360 degrees F.) for a short time, and boiling water they can resist for a long time. Such spores when subsequently placed under favourable conditions will germinate and start bacterial activity anew.

MOTION.

Some species of bacteria have the power of active motion, and may be seen darting rapidly to and fro in the liquid in which they are growing. This motion is produced by flagella which protrude from the body. These flagella (Fig. 15) arise from a membrane surrounding the bacterium, but have an intimate connection with the protoplasmic content. Their distribution is different in different species of bacteria. Some species have a single flagellum at one end (Fig. 15 a). Others have one at each end (Fig. 15 b). Others, again, have, at least just before dividing, a bunch at one or both ends (Fig. 15 c and d), while others, again, have many flagella distributed all over the body in dense profusion (Fig. 15 e). These flagella keep up a lashing to and fro in the liquid, and the lashing serves to propel the bacteria through the liquid.

INTERNAL STRUCTURE.

It is hardly possible to say much about the structure of the bacteria beyond the description of their external forms. With all the variations in detail mentioned, they are extraordinarily simple, and about all that can be seen is their external shape. Of course, they have some internal structure, but we know very little in regard to it. Some microscopists have described certain appearances which they think indicate internal structure. Fig. 16 shows some of these appearances. The matter is as yet very obscure, however. The bacteria appear to have a membranous covering which sometimes is of a cellulose nature. Within it is protoplasm which shows various uncertain appearances. Some microscopists have thought they could find a nucleus, and have regarded bacteria as cells with inclosed nucleii (Figs. 10 a and 15 f). Others have regarded the whole bacterium as a nucleus without any protoplasm, while others, again, have concluded that the discerned internal structure is nothing except an appearance presented by the physical arrangement of the protoplasm. While we may believe that they have some internal structure, we must recognise that as yet microscopists have not been able to make it out. In short, the bacteria after two centuries of study appear to us about as they did at first. They must still be described as minute spheres, rods, or spirals, with no further discernible structure, sometimes motile and sometimes stationary, sometimes producing spores and sometimes not, and multiplying universally by binary fission. With all the development of the modern microscope we can hardly say more than this. Our advance in knowledge of bacteria is connected almost wholly with their methods of growth and the effects they produce in Nature.

ANIMALS OR PLANTS?

There has been in the past not a little question as to whether bacteria should be rightly classed with plants or with animals. They certainly have characters which ally them with both. Their very common power of active independent motion and their common habit of living upon complex bodies for foods are animal characters, and have lent force to the suggestion that they are true animals. But their general form, their method of growth and formation of threads, and their method of spore formation are quite plantlike. Their general form is very similar to a group of low green plants known as Oscillaria. Fig. 17 shows a group of these Oscillariae, and the similarity of this to some of the thread-like bacteria is decided. The Oscillariae are, however, true plants, and are of a green colour. Bacteria are therefore to- day looked upon as a low type of plant which has no chlorophyll, [Footnote: Chlorophyll is the green colouring matter of plants.] but is related to Oscillariae. The absence of the chlorophyll has forced them to adopt new relations to food, and compels them to feed upon complex foods instead of the simple ones, which form the food of green plants. We may have no hesitation, then, in calling them plants. It is interesting to notice that with this idea their place in the organic world is reduced to a small one systematically. They do not form a class by themselves, but are simply a subclass, or even a family, and a family closely related to several other common plants. But the absence of chlorophyll and the resulting peculiar life has brought about a curious anomaly. Whereas their closest allies are known only to botanists, and are of no interest outside of their systematic relations, the bacteria are familiar to every one, and are demanding the life attention of hundreds of investigators. It is their absence of chlorophyll and their consequent dependence upon complex foods which has produced this anomaly.

CLASSIFICATION OF BACTERIA.

While it has generally been recognised that bacteria are plants, any further classification has proved a matter of great difficulty, and bacteriologists find it extremely difficult to devise means of distinguishing species. Their extreme simplicity makes it no easy matter to find points by which any species can be recognised. But in spite of their similarity, there is no doubt that many different species exist. Bacteria which appear to be almost identical, under the microscope prove to have entirely different properties, and must therefore be regarded as distinct species. But how to distinguish them has been a puzzle. Microscopists have come to look upon the differences in shape, multiplication, and formation of spores as furnishing data sufficient to enable them to divide the bacteria into genera. The genus Bacillus, for instance, is the name given to all rod-shaped bacteria which form endogenous spores, etc. But to distinguish smaller subdivisions it has been found necessary to fall back upon other characters, such as the shape of the colony produced in solid gelatine, the power to produce disease, or to oxidize nitrites, etc. Thus at present the different species are distinguished rather by their physiological than their morphological characters. This is an unsatisfactory basis of classification, and has produced much confusion in the attempts to classify bacteria. The problem of determining the species of bacteria is to-day a very difficult one, and with our best methods is still unsatisfactorily solved. A few species of marked character are well known, and their powers of action so well understood that they can be readily recognised; but of the great host of bacteria studied, the large majority have been so slightly experimented upon that their characters are not known, and it is impossible, therefore, to distinguish many of them apart. We find that each bacteriologist working in any special line commonly keeps a list of the bacteria which he finds, with such data in regard to them as he has collected. Such a list is of value to him, but commonly of little value to other bacteriologists from the insufficiency of the data. Thus it happens that a large part of the different species of bacteria described in literature to- day have been found and studied by one investigator alone. By him they have been described and perhaps named. Quite likely the same species may have been found by two or three other bacteriologists, but owing to the difficulty of comparing results and the incompleteness of the descriptions the identity of the species is not discovered, and they are probably described again under different names. The same process may be repeated over and over again, until the same species of bacterium will come to be known by several different names, as it has been studied by different observers.

VARIATION OF BACTERIA.

This matter is made even more confusing by the fact that any species of bacterium may show more or less variation. At one time in the history of bacteriology, a period lasting for many years, it was the prevalent opinion that there was no constancy among bacteria, but that the same species might assume almost any of the various forms and shapes, and possess various properties. Bacteria were regarded by some as stages in the life history of higher plants. This question as to whether bacteria remain constant in character for any considerable length of time has ever been a prominent one with bacteriologists, and even to-day we hardly know what the final answer will be. It has been demonstrated beyond peradventure that some species may change their physiological characters. Disease bacteria, for instance, under certain conditions lose their powers of developing disease. Species which sour milk, or others which turn gelatine green, may lose their characters. Now, since it is upon just such physiological characters as these that we must depend in order to separate different species of bacteria from each other, it will be seen that great confusion and uncertainty will result in our attempts to define species. Further, it has been proved that there is sometimes more or less of a metamorphosis in the life history of certain species of bacteria. The same species may form a short rod, or a long thread, or break up into spherical spores, and thus either a short rod, or a thread, or a spherical form may belong to the same species. Other species may be motile at one time and stationary at another, while at a third period it is a simple mass of spherical spores. A spherical form, when it lengthens before dividing, appears as a short rod, and a short rod form after dividing may be so short as to appear like a spherical organism.

With all these reasons for confusion, it is not to be wondered at that no satisfactory classification of bacteria has been reached, or that different bacteriologists do not agree as to what constitutes a species, or whether two forms are identical or not. But with all the confusion there is slowly being obtained something like system. In spite of the fact that species may vary and show different properties under different conditions, the fundamental constancy of species is everywhere recognised to-day as a fact. The members of the same species may show different properties under different conditions, but it is believed that under identical conditions the properties will be constant. It is no more possible to convert one species into another than it is among the higher orders of plants. It is believed that bacteria do form a group of plants by themselves, and are not to be regarded as stages in the history of higher plants. It is believed that, together with a considerable amount of variability and an occasional somewhat long life history with successive stages, there is also an essential constancy. A systematic classification has been made which is becoming more or less satisfactory. We are constantly learning more and more of the characters, so that they can be recognised in different places by different observers. It is the conviction of all who work with bacteria that, in spite of the difficulties, it is only a matter of time when we shall have a classification and description of bacteria so complete as to characterize the different species accurately.

Even with our present incomplete knowledge of what characterizes a species, it is necessary to use some names. Bacteria are commonly given a generic name based upon their microscopic appearance. There are only a few of these names. Micrococcus, Streptococcus, Staphylococcus, Sarcina, Bacterium, Bacillus, Spirillum, are all the names in common use applying to the ordinary bacteria. There are a few others less commonly used. To this generic name a specific name is commonly added, based upon some physiological character. For example, Bacillus typhosus is the name given to the bacillus which causes typhoid fever. Such names are of great use when the species is a common and well-known one, but of doubtful value for less-known species It frequently happens that a bacteriologist makes a study of the bacteria found in a certain locality, and obtains thus a long list of species hitherto unknown. In these cases it is common simply to number these species rather than name them. This method is frequently advisable, since the bacteriologist can seldom hunt up all bacteriological literature with sufficient accuracy to determine whether some other bacteriologist may not have found the same species in an entirely different locality. One bacteriologist, for example, finds some seventy different species of bacteria in different cheeses. He studies them enough for his own purposes, but not sufficiently to determine whether some other person may not have found the same species perhaps in milk or water. He therefore simply numbers them—a method which conveys no suggestion as to whether they may be new species or not. This method avoids the giving of separate names to the same species found by different observers, and it is hoped that gradually accumulating knowledge will in time group together the forms which are really identical, but which have been described by different observers.

WHERE BACTERIA ARE FOUND.

There are no other plants or animals so universally found in Nature as the bacteria. It is this universal presence, together with their great powers of multiplication, which renders them of so much importance in Nature. They exist almost everywhere on the surface of the earth. They are in the soil, especially at its surface. They do not extend to very great depths of soil, however, few existing below four feet of soil. At the surface they are very abundant, especially if the soil is moist and full of organic material. The number may range from a few hundred to one hundred millions per gramme. [Footnote: One gramme is fifteen grains.] The soil bacteria vary also in species, some two-score different species having been described as common in soil. They are in all bodies of water, both at the surface and below it. They are found at considerable depths in the ocean. All bodies of fresh water contain them, and all sediments in such bodies of water are filled with bacteria. They are in streams of running water in even greater quantity than in standing water. This is simply because running streams are being constantly supplied with water which has been washing the surface of the country and thus carrying off all surface accumulations. Lakes or reservoirs, however, by standing quiet allow the bacteria to settle to the bottom, and the water thus gets somewhat purified. They are in the air, especially in regions of habitation. Their numbers are greatest near the surface of the ground, and decrease in the upper strata of air. Anything which tends to raise dust increases the number of bacteria in the air greatly, and the dust and emanations from the clothes of people crowded in a close room fill the air with bacteria in very great numbers. They are found in excessive abundance in every bit of decaying matter wherever it may be. Manure heaps, dead bodies of animals, decaying trees, filth and slime and muck everywhere are filled with them, for it is in such places that they find their best nourishment. The bodies of animals contain them in the mouth, stomach, and intestine in great numbers, and this is, of course, equally true of man. On the surface of the body they cling in great quantity; attached to the clothes, under the finger nails, among the hairs, in every possible crevice or hiding place in the skin, and in all secretions. They do not, however, occur in the tissues of a healthy individual, either in the blood, muscle, gland, or any other organ. Secretions, such as milk, urine, etc., always contain them, however, since the bacteria do exist in the ducts of the glands which conduct the secretions to the exterior, and thus, while the bacteria are never in the healthy gland itself, they always succeed in contaminating the secretion as it passes to the exterior. Not only higher animals, but the lower animals also have their bodies more or less covered with bacteria. Flies have them on their feet, bees among their hairs, etc.

In short, wherever on the face of Nature there is a lodging place for dust there will be found bacteria. In most of these localities they are dormant, or at least growing only a little. The bacteria clinging to the dry hair can grow but little, if at all, and those in pure water multiply very little. When dried as dust they are entirely dormant. But each individual bacterium or spore has the potential power of multiplication already noticed, and as soon as it by accident falls upon a place where there is food and moisture it will begin to multiply. Everywhere in Nature, then, exists this group of organisms with its almost inconceivable power of multiplication, but a power held in check by lack of food. Furnish them with food and their potential powers become actual. Such food is provided by the dead bodies of animals or plants, or by animal secretions, or from various other sources. The bacteria which are fortunate enough to get furnished with such food material continue to feed upon it until the food supply is exhausted or their growth is checked in some other way. They may be regarded, therefore, as a constant and universal power usually held in check. With their universal presence and their powers of producing chemical changes in food material, they are ever ready to produce changes in the face of Nature, and to these changes we will now turn.



CHAPTER II.

MISCELLANEOUS USE OF BACTERIA IN THE ARTS.

The foods upon which bacteria live are in endless variety, almost every product of animal or vegetable life serving to supply their needs. Some species appear to require somewhat definite kinds of food, and have therefore rather narrow conditions of life, but the majority may live upon a great variety of organic compounds. As they consume the material which serves them as food they produce chemical changes therein. These changes are largely of a nature that the chemist knows as decomposition changes. By this is meant that the bacteria, seizing hold of ingredients which constitute their food, break them to pieces chemically. The molecule of the original food matter is split into simpler molecules, and the food is thus changed in its chemical nature. As a result, the compounds which appear in the decomposing solution are commonly simpler than the original food molecules. Such products are in general called decomposition products, or sometimes cleavage products. Sometimes, however, the bacteria have, in addition to their power of pulling their food to pieces, a further power of building other compounds out of the fragments, thus building up as well as pulling down. But, however they do it, bacteria when growing in any food material have the power of giving rise to numerous products which did not exist in the food mass before. Because of their extraordinary powers of reproduction they are capable of producing these changes very rapidly and can give rise in a short time to large amounts of the peculiar products of their growth.

It is to these powers of producing chemical changes in their food that bacteria owe all their importance in the world. Their power of chemically destroying the food products is in itself of no little importance, but the products which arise as the result of this series of chemical changes are of an importance in the world which we are only just beginning to appreciate. In our attempt to outline the agency which bacteria play in our industries and in natural processes as well, we shall notice that they are sometimes of value simply for their power of producing decomposition; but their greatest value lies in the fact that they are important agents because of the products of their life.

We may notice, in the first place, that in the arts there are several industries which may properly be classed together as maceration industries, all of which are based upon the decomposition powers of bacteria. Hardly any animal or vegetable substance is able to resist their softening influence, and the artisan relies upon this power in several different directions.

BENEFITS DERIVED FROM POWERS OF DECOMPOSITION.

Linen.—Linen consists of certain woody fibres of the stem of the flax. The flax stem is not made up entirely of the valuable fibres, but largely of more brittle wood fibres, which are of no use. The valuable fibres are, however, closely united with the wood and with each other in such an intimate fashion that it is impossible to separate them by any mechanical means. The whole cellular substance of the stem is bound together by some cementing materials which hold it in a compact mass, probably a salt of calcium and pectinic acid. The art of preparing flax is a process of getting rid of the worthless wood fibres and preserving the valuable, longer, tougher, and more valuable fibres, which are then made into linen. But to separate them it is necessary first to soften the whole tissue. This is always done through the aid of bacteria. The flax stems, after proper preparation, are exposed to the action of moisture and heat, which soon develops a rapid bacterial growth. Sometimes this is done by simply exposing the flax to the dew and rain and allowing it to lie thus exposed for some time. By another process the stems are completely immersed in water and allowed to remain for ten to fourteen days. By a third process the water in which the flax is immersed is heated from 75 degrees to 90 degrees F., with the addition of certain chemicals, for some fifty to sixty hours. In all cases the effect is the same. The moisture and the heat cause a growth of bacteria which proceeds with more or less rapidity according to the temperature and other conditions. A putrefactive fermentation is thus set up which softens the gummy substance holding the fibres together. The process is known as "retting," and after it is completed the fibres are easily isolated from each other. A purely mechanical process now easily separates the valuable fibres from the wood fibres. The whole process is a typical fermentation. A disagreeable odour arises from the fermenting flax, and the liquid after the fermentation is filled with products which make valuable manure. The process has not been scientifically studied until very recently. The bacillus which produces the "retting" is known now, however, and it has been shown that the "retting" is a process of decomposition of the pectin cement. No method of separating the linen fibres in the flax from the wood fibres has yet been devised which dispenses with the aid of bacteria.

Jute and Hemp.—Almost exactly the same use is made of bacterial action in the manufacture of jute und hemp. The commercial aspect of the jute industry has grown to be a large one, involving many millions of dollars. Like linen, jute is a fibre of the inner bark of a plant, and is mixed in the bark with a mass of other useless fibrous material. As in the case of linen, a fermentation by bacteria is depended upon as a means of softening the material so that the fibres can be disassociated. The process is called "retting," as in the linen manufacture. The details of the process are somewhat different. The jute is commonly fermented in tanks of stagnant water, although sometimes it is allowed to soak in river water for a sufficient length of time to produce the softening. After the fermentation is thus started the jute fibre is separated from the wood, and is of a sufficient flexibility and toughness to be woven into sacking, carpets, curtains, table covers, and other coarse cloth.

Practically the same method is used in separating the tough fibres of the hemp. The hemp plant contains some long flexible fibres with others of no value, and bacterial fermentation is relied upon to soften the tissues so that they may be separated.

Cocoanut fibre, a somewhat similar material is obtained from the husk of the cocoanut by the same means. The unripened husk is allowed to steep and ferment in water for a long time, six months or a year being required. By this time the husk has become so softened that it can be beaten until the fibres separate and can be removed. They are subsequently made into a number of coarse articles, especially valuable for their toughness. Door mats, brushes, ships' fenders, etc., are illustrations of the sort of articles made from them.

In each of these processes the fermentation must have a tendency to soften the desired fibres as well as the connecting substance. Putrefaction attacks all kinds of vegetable tissue, and if this "retting" continues too long the desired fibre is decidedly injured by the softening effect of the fermentation. It is quite probable that, even as commonly carried on, the fermentation has some slight injurious effect upon the fibre, and that if some purely mechanical means could be devised for separating the fibre from the wood it would produce a better material. But such mechanical means has not been devised, and at present a putrefactive fermentation appears to be the only practical method of separating the fibres.

Sponges.—A somewhat similar use is made of bacteria in the commercial preparation of sponges. The sponge of commerce is simply the fibrous skeleton of a marine animal. When it is alive this skeleton is completely filled with the softer parts of the animal, and to fit the sponge for use this softer organic material must be got rid of. It is easily accomplished by rotting. The fresh sponges are allowed to stand in the warm sun and very rapidly decay. Bacteria make their way into the sponge and thoroughly decompose the soft tissues. After a short putrefaction of this sort the softened organic matter can be easily washed out of the skeleton and leave the clean fibre ready for market.

Leather preparation.—The tanning of leather is a purely chemical process, and in some processes the whole operation of preparing the leather is a chemical one. In others, however, especially in America, bacteria are brought into action at one stage. The dried hide which comes to the tannery must first have the hair removed together with the outer skin. The hide for this purpose must be moistened and softened. In some tanneries this is done by steeping it in chemicals. In others, however, it is put into water and slightly heated until fermentation arises. The fermentation softens it so that the outer skin can be easily removed with a knife, and the removal of hair is accomplished at the same time. Bacterial putrefaction in the tannery is thus an assistance in preparing the skin for the tanning proper. Even in the subsequent tanning a bacterial fermentation appears to play a part, but little is yet known in regard to it.

Maceration of skeletons.—The making of skeletons for museums and anatomical instruction in general is no very great industry, and yet it is one of importance. In the making of skeletons the process of maceration is commonly used as an aid. The maceration consists simply in allowing the skeleton to soak in water for a day or two after cleaning away the bulk of the muscles. The putrefaction that arises softens the connective tissues so much that the bones may be readily cleaned of flesh.

Citric acid.—Bacterial fermentation is employed also in the ordinary preparation of citric acid. The acid is made chiefly from the juice of the lemon. The juice is pressed from the fruit and then allowed to ferment. The fermentation aids in separating a mucilaginous mass and making it thus possible to obtain the citric acid in a purer condition. The action is probably similar to the maceration processes described above, although it has not as yet been studied by bacteriologists.

BENEFITS DERIVED FROM THE PRODUCTS OF BACTERIAL LIFE.

While bacteria thus play a part in our industries simply from their power of producing decomposition, it is primarily because of the products of their action that they are of value. Wherever bacteria seize hold of organic matter and feed upon it, there are certain to be developed new chemical compounds, resulting largely from decomposition, but partly also from constructive processes. These new compounds are of great variety. Different species of bacteria do not by any means produce the same compounds even when growing in and decomposing the same food material. Moreover, the same species of bacteria may give rise to different products when growing in different food materials. Some of the compounds produced by such processes are poisonous, others are harmless. Some are gaseous, others are liquids. Some have peculiar odours, as may be recognised from the smell arising from a bit of decaying meat. Others have peculiar tastes, as may be realized in the gamy taste of meat which is in the incipient stages of putrefaction. By purely empirical means mankind has learned methods of encouraging the development of some of these products, and is to-day making practical use of this power, possessed by bacteria, of furnishing desired chemical compounds. Industries involving the investment of hundreds of millions of dollars are founded upon the products of bacterial life, and they have a far more important relation to our everyday life than is commonly imagined. In many cases the artisan who is dependent upon this action of microscopic life is unaware of the fact. His processes are those which experience has taught produce desired results, but, nevertheless, his dependence upon bacteria is none the less fundamental.

BACTERIA IN THE FERMENTATIVE INDUSTRIES.

We may notice, first, several miscellaneous instances of the application of bacteria to various fermentative industries where their aid is of more or less value to man. In some of the examples to be mentioned the influence of bacteria is profound and fundamental, while in others it is only incidental. The fermentative industries of civilization are gigantic in extent, and have come to be an important factor in modern civilized life. The large part of the fermentation is based upon the growth of a class of microscopic plants which we call yeasts. Bacteria and yeasts are both microscopic plants, and perhaps somewhat closely related to each other. The botanist finds a difference between them, based upon their method of multiplication, and therefore places them in different classes (Fig. 2, page 19). In their general power of producing chemical changes in their food products, yeasts agree closely with bacteria, though the kinds of chemical changes are different. The whole of the great fermentative industries, in which are invested hundreds of millions of dollars, is based upon chemical decompositions produced by microscopic plants. In the great part of commercial fermentations alcohol is the product desired, and alcohol, though it is sometimes produced by bacteria, is in commercial quantities produced only by yeasts. Hence it is that, although the fermentations produced by bacteria are more common in Nature than those produced by yeasts and give rise to a much larger number of decomposition products, still their commercial aspect is decidedly less important than that of yeasts. Nevertheless, bacteria are not without their importance in the ordinary fermentative processes. Although they are of no importance as aids in the common fermentative processes, they are not infrequently the cause of much trouble. In the fermentation of malt to produce beer, or grape juice to produce wine, it is the desire of the brewer and vintner to have this fermentation produced by pure yeasts, unmixed with bacteria. If the yeast is pure the fermentation is uniform and successful. But the brewer and vintner have long known that the fermentation is frequently interfered with by irregularities. The troubles which arise have long been known, but the bacteriologist has finally discovered their cause, and in general their remedy. The cause of the chief troubles which arise in the fermentation is the presence of contaminating bacteria among the yeasts. These bacteria have been more or less carefully studied by bacteriologists, and their effect upon the beer or wine determined. Some of them produce acid and render the products sour; others make them bitter; others, again, produce a slimy material which makes the wine or beer "ropy." Something like a score of bacteria species have been found liable to occur in the fermenting material and destroy the value of the product of both the wine maker and the beer brewer. The species of bacteria which infect and injure wine are different from those which infect and injure beer. They are ever present as possibilities in the great alcoholic fermentations. They are dangers which must be guarded against. In former years the troubles from these sources were much greater than they are at present. Since it has been demonstrated that the different imperfections in the fermentative process are due to bacterial impurities, commonly in the yeasts which are used to produce the fermentation, methods of avoiding them are readily devised. To-day the vintner has ready command of processes for avoiding the troubles which arise from bacteria, and the brewer is always provided with a microscope to show him the presence or absence of the contaminating bacteria. While, then, the alcoholic fermentations are not dependent upon bacteria, the proper management of these fermentations requires a knowledge of their habits and characters.

There are certain other fermentative processes of more or less importance in their commercial aspects, which are directly dependent upon bacterial action, Some of them we should unhesitatingly look upon as fermentations, while others would hardly be thought of as belonging to the fermentation industries.

VINEGAR.

The commercial importance of the manufacture of vinegar, though large, does not, of course, compare in extent with that of the alcoholic fermentations. Vinegar is a weak solution of acetic acid, together with various other ingredients which have come from the materials furnishing the acid. In the manufacture of vinegar, alcohol is always used as the source of the acetic acid. The production of acetic acid from alcohol is a simple oxidation. The equation C2H6O + O2 = C2H4O2 + H2O shows the chemical change that occurs. This oxidation can be brought about by purely chemical means. While alcohol will not readily unite with oxygen under common conditions, if the alcohol is allowed to pass over a bit of platinum sponge the union readily occurs and acetic acid results. This method of acetic-acid production is possible experimentally, but is impracticable on any large scale. In the ordinary manufacture of vinegar the oxidation is a true fermentation, and brought about by the growth of bacteria.

In the commercial manufacture of vinegar several different weak alcoholic solutions are used. The most common of these are fermented malt, weak wine, cider, and sometimes a weak solution of spirit to which is added sugar and malt. If these solutions are allowed to stand for a time in contact with air, they slowly turn sour by the gradual conversion of the alcohol into acetic acid. At the close of the process practically all of the alcohol has disappeared. Ordinarily, however, not all of it has been converted into acetic acid, for the oxidation does not all stop at this step. As the oxidation goes on, some of the acid is oxidized into carbonic dioxide, which is, of course, dissipated at once into the air, and if the process is allowed to continue unchecked for a long enough period much of the acetic acid will be lost in this way.

The oxidation of the alcohol in all commercial production of vinegar is brought about by the growth of bacteria in the liquid. When the vinegar production is going on properly, there is formed on the top of the liquid a dense felted mass known as the "mother of vinegar." This mass proves to be made of bacteria which have the power of absorbing oxygen from the air, or, at all events, of causing the alcohol to unite with oxygen. It was at first thought that a single species of bacterium was thus the cause of the oxidation of alcohol, and this was named Mycoderma aceti. But further study has shown that several have the power, and that even in the commercial manufacture of vinegar several species play a part (Fig. 18), although the different species are not yet very thoroughly studied. Each appears to act best under different conditions. Some of them act slowly, and others rapidly, the slow- growing species appearing to produce the larger amount of acid in the end. After the amount of acetic acid reaches a certain percentage, the bacteria are unable to produce more, even though there be alcohol still left unoxidized. A percentage as high as fourteen per cent, commonly destroys all their power of growth. The production of the acid is wholly dependent upon the growth of the bacteria, and the secret of the successful vinegar manufacture is the skilful manipulation of these bacteria so as to keep them in the purest condition and to give them the best opportunity for growth.

One method of vinegar manufacture which is quite rapid is carried on in a slightly different manner. A tall cylindrical chamber is filled with wood shavings, and a weak solution of alcohol is allowed to trickle slowly through it. The liquid after passing over the shavings comes out after a number of hours well charged with acetic acid. This process at first sight appears to be a purely chemical one, and reminds us of the oxidation which occurs when alcohol is allowed to pass over a platinum sponge. It has been claimed, indeed, that this is a chemical oxidation in which bacteria play no part. But this appears to be an error. It is always found necessary in this method to start the process by pouring upon the shavings some warm vinegar. Unless in this way the shavings become charged with the vinegar-holding bacteria the alcohol will not undergo oxidation during its passage over them, and after the bacteria thus introduced have grown enough to coat the shavings thoroughly the acetic-acid production is much more rapid than at first. If vinegar is allowed to trickle slowly down a suspended string, so that its bacteria may distribute themselves through the string, and then alcohol be allowed to trickle over it in the same way, the oxidation takes place and acetic acid is formed. From the accumulation of such facts it has come to be recognised that all processes for the commercial manufacture of vinegar depend upon the action of bacteria. While the oxidation of alcohol into acetic acid may take place by purely chemical means, these processes are not practical on a large scale, and vinegar manufacturers everywhere depend upon bacteria as their agents in producing the oxidation. These bacteria, several species in all, feed upon the nitrogenous matter in the fermenting mass and produce the desired change in the alcohol.

This vinegar fermentation is subject to certain irregularities, and the vinegar manufacturers can not always depend upon its occurring in a satisfactory manner. Just as in brewing, so here, contaminating bacteria sometimes find their way into the fermenting mass and interfere with its normal course. In particular, the flavour of the vinegar is liable to suffer from such causes. As yet our vinegar manufacturers have not applied to acetic fermentation the same principle which has been so successful in brewing—namely, the use, as a starter of the fermentation, of a pure culture of the proper species of bacteria. This has been done experimentally and proves to be feasible. In practice, however, vinegar makers find that simpler methods of obtaining a starter—by means of which they procure a culture nearly though not absolutely pure—are perfectly satisfactory. It is uncertain whether really pure cultures will ever be used in this industry.

LACTIC ACID.

The manufacture of lactic acid is an industry of less extent than that of acetic acid, and yet it is one which has some considerable commercial importance. Lactic acid is used in no large quantity, although it is of some value as a medicine and in the arts. For its production we are wholly dependent upon bacteria. It is this acid which, as we shall see, is produced in the ordinary souring of milk, and a large number of species of bacteria are capable of producing the acid from milk sugar. Any sample of sour milk may therefore always be depended upon to contain plenty of lactic organisms. In its manufacture for commercial purposes milk is sometimes used as a source, but more commonly other substances. Sometimes a mixture of cane sugar and tartaric acid is used. To start the fermentation the mixture is inoculated with a mass of sour milk or decaying cheese, or both, such a mixture always containing lactic organisms. To be sure, it also contains many other bacteria which have different effects, but the acid producers are always so abundant and grow so vigorously that the lactic fermentation occurs in spite of all other bacteria. Here also there is a possibility of an improvement in the process by the use of pure cultures of lactic organisms. Up to the present, however, there has been no application of such methods. The commercial aspects of the industry are not upon a sufficiently large scale to call for much in this direction.

At the present time the only method we have for the manufacture of lactic acid is dependent upon bacteria. Chemical processes for its manufacture are known, but not employed commercially. There are several different kinds of lactic acid. They differ from each other in the relations of the atoms within their molecule, and in their relation to polarized light, some forms rotating the plane of polarized light to the right, others to the left, while others are inactive in this respect. All the types are produced by fermentation processes, different species of bacteria having powers of producing the different types.

BUTYRIC ACID.

Butyric acid is another acid for which we are chiefly dependent upon bacteria. This acid is of no very great importance, and its manufacture can hardly be called an industry; still it is to a certain extent made, and is an article of commerce. It is an acid that can be manufactured by chemical means, but, as in the case of the last two acids, its commercial manufacture is based upon bacterial action. Quite a number of species of bacteria can produce butyric acid, and they produce it from a variety of different sources. Butyric acid is a common ingredient in old milk and in butter, and its formation by bacteria was historically one of the first bacterial fermentations to be clearly understood. It can be produced also in various sugar and starchy solutions. Glycerine may also undergo a butyric fermentation. The presence of this acid is occasionally troublesome, since it is one of the factors in the rancidity of butter and other similar materials.

INDIGO PREPARATION.

The preparation of indigo from the indigo plant is a fermentative process brought about by a specific bacterium. The leaves of the plant are immersed in water in a large vat, and a rapid fermentation arises. As a result of the fermentation the part of the plant which is the basis of the indigo is separated from the leaves and dissolved in the water; and as a second feature of the fermentation the soluble material is changed in its chemical nature into indigo proper. As this change occurs the characteristic blue colour is developed, and the material is rendered insoluble in water. It therefore makes its appearance as a blue mass separated from the water, and is then removed as indigo.

Of the nature of the process we as yet know very little. That it is a fermentation is certain, and it has been proved that it is produced by a definite species of bacterium which occurs on the indigo leaves. If the sterilized leaves are placed in sterile water no fermentation occurs and no indigo is formed. If, however, some of the specific bacteria are added to the mass the fermentation soon begins and the blue colour of the indigo makes its appearance. It is plain, therefore, that indigo is a product of bacterial fermentation, and commonly due to a single definite species of bacterium. Of the details of the formation, however, we as yet know little, and no practical application of the facts have yet been made.

BACTERIA IN TOBACCO CURING.

A fermentative process of quite a different nature, but of immense commercial value, is found in the preparation of tobacco. The process by which tobacco is prepared is a long and somewhat complicated one, consisting of a number of different stages. The tobacco, after being first dried in a careful manner, is subsequently allowed to absorb moisture from the atmosphere, and is then placed in large heaps to undergo a further change. This process appears to be a fermentation, for the temperature of the mass rises rapidly, and every indication of a fermentative action is seen. The tobacco in these heaps is changed occasionally, the heap being thrown down and built up again in such a way that the portion which was first at the bottom comes to the top, and in this way all parts of the heap may become equally affected by the process. After this process the tobacco is sent to the different manufacturers, who finish the process of curing. The further treatment it receives varies widely according to the desired product, whether for smoking or for snuff, etc. In all cases, however, fermentations play a prominent part. Sometimes the leaves are directly inoculated with fermenting material. In the preparation of snuff the details of the process are more complicated than in the preparation of smoking tobacco. The tobacco, after being ground and mixed with certain ingredients, is allowed to undergo a fermentation which lasts for weeks, and indeed for months. In the different methods of preparing snuff the fermentations take place in different ways, and sometimes the tobacco is subjected to two or three different fermentative actions. The result of the whole is the slow preparation of the commercial product. It is during the final fermentative processes that the peculiar colour and flavour of the snuff are developed, and it is during the fermentation of the leaves of the smoking tobacco—either the original fermentation or the subsequent ones— that the special flavours and aromas of tobacco are produced.

It can not be claimed for a moment that these changes by which the tobacco is cured and finally brought to a marketable condition are due wholly to bacteria. There is no question that chemical and physical phenomena play an important part in them. Nevertheless, from the moment when the tobacco is cut in the fields until the time it is ready for market the curing is very intimately associated with bacteria and fermentative organisms in general. Some of these processes are wholly brought about by bacterial life; in others the micro-organisms aid the process, though they perhaps can not be regarded as the sole agents.

At the outset the tobacco producer has to contend with a number of micro-organisms which may produce diseases in his tobacco. During the drying process, if the temperature or the amount of moisture or the access of air is not kept in a proper condition, various troubles arise and various diseases make their appearance, which either injure or ruin the value of the product. These appear to be produced by micro-organisms of different sorts. During the fermentation which follows the drying the producer has to contend with micro-organisms that are troublesome to him; for unless the phenomena are properly regulated the fermentation that occurs produces effects upon the tobacco which ruin its character. From the time the tobacco is cut until the final stage in the curing the persons engaged in preparing it for market must be on a constant watch to prevent the growth within it of undesirable organisms. The preparation of tobacco is for this reason a delicate operation, and one that will be very likely to fail unless the greatest care is taken. In the several fermentative processes which occur in the preparation there is no question that micro-organisms aid the tobacco producer and manufacturer. Bacteria produce the first fermentation that follows the drying, and it is these organisms too, in large measure, that give rise to all the subsequent fermentations, although seemingly in some cases purely chemical processes materially aid. Now the special quality of the tobacco is in part dependent upon the peculiar type of fermentation which occurs in one or another of these fermenting actions. It is the fermentation that gives rise to the peculiar flavour and to the aroma of the different grades of tobacco. Inasmuch as the various flavours which characterize tobacco of different grades are developed, at least to a large extent, during the fermentation processes, it is a natural supposition that the different qualities of the tobacco, so far as concerns flavour, are due to the different types of fermentation. The number of species of bacteria which are found upon the tobacco leaves in the various stages of its preparation is quite large, and from what we have already learned it is inevitable that the different kinds of bacteria will produce different results in the fermenting process. It would seem natural, therefore, to assume that the different flavours of different grades may not unlikely be due to the fact that the tobacco in the different cases has been fermented under the influence of different kinds of bacteria.

Nor is this simply a matter of inference. To a certain extent experimental evidence has borne out the conclusion, and has given at least a slight indication of practical results in the future. Acting upon the suggestion that the difference between the high grades of tobacco and the poorer grades is due to the character of the bacteria that produce the fermentation, certain bacteriologists have attempted to obtain from a high quality of tobacco the species of bacteria which are infesting it. These bacteria have then been cultivated by bacteriological methods and used in experiments for the fermentation of tobacco. If it is true that the flavour of high grade tobacco is in large measure, or even in part, due to the action of the peculiar microbes from the soil where it grows, it ought to be possible to produce similar flavours in the leaves of tobacco grown in other localities, if the fermentation of the leaves is carried on by means of the pure cultures of bacteria obtained from the high grade tobacco. Not very much has been done or is known in this connection as yet. Two bacteriologists have experimented independently in fermenting tobacco leaves by the action of pure cultures of bacteria obtained from such sources. Each of them reports successful experiments. Each claims that they have been able to improve the quality of tobacco by inoculating the leaves with a pure culture of bacteria obtained from tobacco having high quality in flavour. In addition to this, several other bacteriologists have carried on experiments sufficient to indicate that the flavours of the tobacco and the character of the ripening may be decidedly changed by the use of different species of micro-organisms in the fermentations that go on during the curing processes.

In regard to the whole matter, however, we must recognise that as yet we have very little knowledge. The subject has been under investigation for only a short time; and, while considerable information has been derived, this information is not thoroughly understood, and our knowledge in regard to the matter is as yet in rather a chaotic condition. It seems certain, however, that the quality of tobacco is in large measure dependent upon the character of the fermentations that occur at different stages of the curing. It seems certain also that these fermentations are wholly or chiefly produced by microorganisms, and that the character of the fermentation is in large measure dependent upon the species of micro-organisms that produce it. If these are facts, it would seem not improbable that a further study may produce practical results for this great industry. The study of yeasts and the methods of keeping yeast from contaminations has revolutionised the brewing industry. Perhaps in this other fermentative industry, which is of such great commercial extent, the use of pure cultures of bacteria may in the future produce as great revolutions in methods as it has in the industry of the alcoholic fermentation.

It must not, however, be inferred that the differences in grades of tobacco grown in different parts of the world are due solely to variations in the curing processes and to the types of fermentation. There are differences in the texture of the leaves, differences in the chemical composition of the tobaccoes, which are due undoubtedly to the soils and the climatic conditions in which they grow, and these, of course, will never be affected by changing the character of the ferment active processes. It is, however, probable that in so far as the flavours that distinguish the high and low grades of tobacco are due to the character of the fermentative processes, they may be in the future, at least to a large extent, controlled by the use of pure cultures in curing processes. Seemingly, then, there is as great a future in the development of this fermentative industry as there has been in the past in the development of the fermentative industry associated with brewing and vinting.

OPIUM.

Opium for smoking purposes is commonly allowed to undergo a curing process which lasts several months. This appears to be somewhat similar to the curing of tobacco. Apparently it is a fermentation due to the growth of microorganisms. The organisms in question are not, however, bacteria in this case, but a species of allied fungus. The plant is a mould, and it is claimed that inoculation of the opium with cultures of this mould hastens the curing.

TROUBLESOME FERMENTATIONS.

Before leaving this branch of the subject it is necessary to notice some of the troublesome fermentations which are ever interfering with our industries, requiring special methods, or, indeed, sometimes developing special industries to meet them. As agents of decomposition, bacteria will of course be a trouble whenever they get into material which it is desired to preserve. Since they are abundant everywhere, it is necessary to count upon their attacking with certainty any fermentable substance which is exposed to air and water. Hence they are frequently the cause of much trouble. In the fermentative industries they occasionally cause an improper sort of fermentation to occur unless care is taken to prevent undesired species of bacteria from being present. In vinegar making, improper species of bacteria obtaining access to the solution give rise to undesirable flavours, greatly injuring the product. In tobacco curing it is very common for the wrong species of bacteria to gain access to the tobacco at some stage of the curing and by their growth give rise to various troubles. It is the ubiquitous presence of bacteria which makes it impossible to preserve fruits, meats, or vegetables for any length of time without special methods. This fact in itself has caused the development of one of our most important industries. Canning meats or fruits consists in nothing more than bringing them into a condition in which they will be preserved from attack of these micro-organisms. The method is extremely simple in theory. It is nothing more than heating the material to be preserved to a high temperature and then sealing it hermetically while it is still hot. The heat kills all the bacteria which may chance to be lodged in it, and the hermetical sealing prevents other bacteria from obtaining access. Inasmuch as all organic decomposition is produced by bacterial growth, such sterilized and sealed material will be preserved indefinitely when the operation is performed carefully enough. The methods of accomplishing this with sufficient care are somewhat varied in different industries, but they are all fundamentally the same. It is an interesting fact that this method of preserving meats was devised in the last century, before the relation of micro-organisms to fermentation and putrefaction was really suspected. For a long time it had been in practical use while scientists were still disputing whether putrefaction could be avoided by preventing the access of bacteria. The industry has, however, developed wonderfully within the last few years, since the principles underlying it have been understood. This understanding has led to better methods of destroying bacterial life and to proper sealing, and these have of course led to greater success in the preservation, until to-day the canning industries are among those which involve capital reckoned in the millions.