Transcriber's note: A few typographical errors have been corrected: they are listed at the end of the text. Volume and page numbers have been incorporated into the text of each page as: v.03 p.0001.
ā signifies "a with macron"; ḥ "h with dot below"; š "s with caron"; and so forth. In the article BALLISTICS, [Integral,a:b] or [Sum,a:b] indicates a definite integral or a summation between lower limit a and upper limit b. [Integral] by itself indicates an indefinite integral. [=3].6090480 etc. denote a barred digit in logarithms.
Musical pitches are expressed in Acoustical Society of America notation: C4 is middle C, B3 the tone below.
THE ENCYCLOPAEDIA BRITANNICA
A DICTIONARY OF ARTS, SCIENCES, LITERATURE AND GENERAL INFORMATION
AUSTRIA LOWER to BISECTRIX
[E-Text Edition of Volume III - Part 1 of 2, Slice 2 of 3 - BACONTHORPE to BANKRUPTCY]
* * * * *
BACONTHORPE [BACON, BACO, BACCONIUS], JOHN (d. 1346), known as "the Resolute Doctor," a learned Carmelite monk, was born at Baconthorpe in Norfolk. He seems to have been the grandnephew of Roger Bacon (Brit. Mus. Add. MS. 19. 116). Brought up in the Carmelite monastery of Blakeney, near Walsingham, he studied at Oxford and Paris, where he was known as "Princeps" of the Averroists. Renan, however, says that he merely tried to justify Averroism against the charge of heterodoxy. In 1329 he was chosen twelfth provincial of the English Carmelites. He appears to have anticipated Wycliffe in advocating the subordination of the clergy to the king. In 1333 he was sent for to Rome, where, we are told, he first maintained the pope's authority in cases of divorce; but this opinion he retracted. He died in London in 1346. His chief work, Doctoris resoluti Joannis Bacconis Anglici Carmelitae radiantissimi opus super quattuor sententiarum libris (published 1510), has passed through several editions. Nearly three centuries later, it was still studied at Padua, the last home of Averroism, and Lucilio Vanini speaks of him with great veneration.
See Brucker, Hist. Crit. iii. 865; Stoeckl, Phil. d. Mittel. ii. 1044-1045; Haureau, Phil. Scol. ii. 476; K. Prantl, Ges. d. Logik, iii. 318. For information as to his life, not found otherwise and of doubtful accuracy, see J. B. de Lezana's Annales Sacri, iv.
BACSANYI, JANOS (1763-1845), Hungarian poet, was born at Tapolcza on the 11th of May 1763. In 1785 he published his first work, a patriotic poem, The Valour of the Magyars. In the same year he obtained a situation as clerk in the treasury at Kaschau, and there, in conjunction with other two Hungarian patriots, edited the Magyar Museum, which was suppressed by the government in 1792. In the following year he was deprived of his clerkship; and in 1794, having taken part in the conspiracy of Bishop Martinovich, he was thrown into the state prison of the Spielberg, near Bruenn, where he remained for two years. After his release he took a considerable share in the Magyar Minerva, a literary review, and then proceeded to Vienna, where he obtained a post in the bank, and married. In 1809 he translated Napoleon's proclamation to the Magyars, and, in consequence of this anti-Austrian act, had to take refuge in Paris. After the fall of Napoleon he was given up to the Austrians, who allowed him to reside at Linz, on condition of never leaving that town. He published a collection of poems at Pest, 1827 (2nd ed. Buda, 1835), and also edited the poetical works of Anyos and Faludi. He died at Linz on the 12th of May 1845.
BACTERIOLOGY. The minute organisms which are commonly called "bacteria" are also known popularly under other designations, e.g. "microbes," "micro-organisms," "microphytes," "bacilli," "micrococci." All these terms, including the usual one of bacteria, are unsatisfactory; for "bacterium," "bacillus" and "micrococcus" have narrow technical meanings, and the other terms are too vague to be scientific. The most satisfactory designation is that proposed by Naegeli in 1857, namely "schizomycetes," and it is by this term that they are usually known among botanists; the less exact term, however, is also used and is retained in this article since the science is commonly known as "bacteriology." The first part of this article deals with the general scientific aspects of the subject, while a second part is concerned with the medical aspects.
I. THE STUDY OF BACTERIA
The general advances which have been made of late years in the study of bacteria are clearly brought to mind when we reflect that in the middle of the 19th century these organisms were only known to a few experts and in a few forms as curiosities of the microscope, chiefly interesting for their minuteness and motility. They were then known under the name of "animalculae," and were confounded with all kinds of other small organisms. At that time nothing was known of their life-history, and no one dreamed of their being of importance to man and other living beings, or of their capacity to produce the profound chemical changes with which we are now so familiar. At the present day, however, not only have hundreds of forms or species been described, but our knowledge of their biology has so extended that we have entire laboratories equipped for their study, and large libraries devoted solely to this subject. Furthermore, this branch of science has become so complex that the bacteriological departments of medicine, of agriculture, of sewage, &c., have become more or less separate studies.
The schizomycetes or bacteria are minute vegetable organisms devoid of chlorophyll and multiplying by repeated bipartitions. They consist of single cells, which may be spherical, oblong or cylindrical in shape, or of filamentous or other aggregates of cells. They are characterized by the absence of ordinary sexual reproduction and by the absence of an ordinary nucleus. In the two last-mentioned characters and in their manner of division the bacteria resemble Schizophyceae (Cyanophyceae or blue-green algae), and the two groups of Schizophyceae and Schizomycetes are usually united in the class Schizophyta, to indicate the generally received view that most of the typical bacteria have been derived from the Cyanophyceae. Some forms, however, such as "Sarcina," have their algal analogues in Palmellaceae among the green algae, while Thaxter's group of Myxobacteriaceae suggests a relationship with the Myxomycetes. The existence of ciliated micrococci together with the formation of endospores—structures not known in the Cyanophyceae—reminds us of the flagellate Protozoa, e.g. Monas, Chromulina. Resemblances also exist between the endospores and the spore-formations in the Saccharomycetes, and if Bacillus inflatus, B. ventriculus, &c., really form more than one spore in the cell, these analogies are strengthened. Schizomycetes such as Clostridium, Plectridium, &c., where the sporiferous cells enlarge, bear out the same argument, and we must not forget that there are extremely minute "yeasts," easily mistaken for Micrococci, and that yeasts occasionally form only one spore in the cell.
Nor must we overlook the possibility that the endospore-formation in non-motile bacteria more than merely resembles the development of azygospores in the Conjugatae, and some Ulothricaceae, if reduced in size, would resemble them. Meyer regards them as chlamydospores, and Klebs as "carpospores" or possibly chlamydospores similar to the endospores of yeast. [v.03 p.0157] The former also looks on the ordinary disjointing bacterial cell as an oidium, and it must be admitted that since Brefeld's discovery of the frequency of minute oidia and chlamydospores among the fungi, the probability that some so-called bacteria—and this applies especially to the branching forms accepted by some bacteriologists—are merely reduced fungi is increased. Even the curious one-sided growth of certain species which form sheaths and stalks—e.g. Bacterium vermiforme, B. pediculatum—can be matched by Algae such as Oocardium, Hydrurus, and some Diatoms. It is clear then that the bacteria are very possibly a heterogeneous group, and in the present state of our knowledge their phylogeny must be considered as very doubtful.
Nearly all bacteria, owing to the absence of chlorophyll, are saprophytic or parasitic forms. Most of them are colourless, but a few secrete colouring matters other than chlorophyll. In size their cells are commonly about 0.001 mm. (1 micromillimetre or 1 [micron]) in diameter, and from two to five times that length, but smaller ones and a few larger ones are known. Some of the shapes assumed by the cells are shown in fig. 1.
[Sidenote: Distribution in Time.]
That bacteria have existed from very early periods is clear from their presence in fossils; and although we cannot accept all the conclusions drawn from the imperfect records of the rocks, and may dismiss as absurd the statements that geologically immured forms have been found still living, the researches of Renault and van Tieghem have shown pretty clearly that large numbers of bacteria existed in Carboniferous and Devonian times, and probably earlier.
[Sidenote: Distribution in Space.]
Schizomycetes are ubiquitous as saprophytes in still ponds and ditches, in running streams and rivers, and in the sea, and especially in drains, bogs, refuse heaps, and in the soil, and wherever organic infusions are allowed to stand for a short time. Any liquid (blood, urine, milk, beer, &c.) containing organic matter, or any solid food-stuff (meat preserves, vegetables, &c.), allowed to stand exposed to the air soon swarms with bacteria, if moisture is present and the temperature not abnormal. Though they occur all the world over in the space, air and on the surface of exposed bodies, it is not to be supposed that they are by any means equally distributed, and it is questionable whether the bacteria suspended in the air ever exist in such enormous quantities as was once believed. The evidence to hand shows that on heights and in open country, especially in the north, there may be few or even no Schizomycetes detected in the air, and even in towns their distribution varies greatly; sometimes they appear to exist in minute clouds, as it were, with interspaces devoid of any, but in laboratories and closed spaces where their cultivation has been promoted the air may be considerably laden with them. Of course the distribution of bodies so light and small is easily influenced by movements, rain, wind, changes of temperature, &c. As parasites, certain Schizomycetes inhabit and prey upon the organs of man and animals in varying degrees, and the conditions for their growth and distribution are then very complex. Plants appear to be less subject to their attacks—possibly, as has been suggested, because the acid fluids of the higher vegetable organisms are less suited for the development of Schizomycetes; nevertheless some are known to be parasitic on plants. Schizomycetes exist in every part of the alimentary canal of animals, except, perhaps, where acid secretions prevail; these are by no means necessarily harmful, though, by destroying the teeth for instance, certain forms may incidentally be the forerunners of damage which they do not directly cause.
Little was known about these extremely minute organisms before 1860. A. van Leeuwenhoek figured bacteria as far back as the 17th century, and O. F. Mueller knew several important forms in 1773, while Ehrenberg in 1830 had advanced to the commencement of a scientific separation and grouping of them, and in 1838 had proposed at least sixteen species, distributing them into four genera. Our modern more accurate though still fragmentary knowledge of the forms of Schizomycetes, however, dates from F. J. Conn's brilliant researches, the chief results of which were published at various periods between 1853 and 1872; Cohn's classification of the bacteria, published in 1872 and extended in 1875, has in fact dominated the study of these organisms almost ever since. He proceeded in the main on the assumption that the forms of bacteria as met with and described by him are practically constant, at any rate within limits which are not wide: observing that a minute spherical micrococcus or a rod-like bacillus regularly produced similar micrococci and bacilli respectively, he based his classification on what may be considered the constancy of forms which he called species and genera. As to the constancy of form, however, Cohn maintained certain reservations which have been ignored by some of his followers. The fact that Schizomycetes produce spores appeals to have been discovered by Cohn in 1857, though it was expressed dubiously in 1872; these spores had no doubt been observed previously. In 1876, however, Cohn had seen the spores germinate, and Koch, Brefeld, Pratzmowski, van Tieghem, de Bary and others confirmed the discovery in various species.
The supposed constancy of forms in Cohn's species and genera received a shock when Lankester in 1873 pointed out that his Bacterium rubescens (since named Beggiatoa roseo-persicina, Zopf) passes through conditions which would have been described by most observers influenced by the current doctrine as so many separate "species" or even "genera,"—that in fact forms known as Bacterium, Micrococcus, Bacillus, Leptothrix, &c., occur as phases in one life-history. Lister put forth similar ideas about the same time; and Billroth came forward in 1874 with the extravagant view that the various bacteria are only different states of one and the same organism which he called Cocco-bacteria septica. From that time the question of the pleomorphism (mutability of shape) of the bacteria has been hotly discussed: but it is now generally agreed that, while a [v.03 p.0158] certain number of forms may show different types of cell during the various phases of the life-history, yet the majority of forms are uniform, showing one type of cell throughout their life-history. The question of species in the bacteria is essentially the same as in other groups of plants; before a form can be placed in a satisfactory classificatory position its whole life-history must be studied, so that all the phases may be known. In the meantime, while various observers were building up our knowledge of the morphology of bacteria, others were laying the foundation of what is known of the relations of these organisms to fermentation and disease—that ancient will-o'-the-wisp "spontaneous generation" being revived by the way. When Pasteur in 1857 showed that the lactic fermentation depends on the presence of an organism, it was already known from the researches of Schwann (1837) and Helmholtz (1843) that fermentation and putrefaction are intimately connected with the presence of organisms derived from the air, and that the preservation of putrescible substances depends on this principle. In 1862 Pasteur placed it beyond reasonable doubt that the ammoniacal fermentation of urea is due to the action of a minute Schizomycete; in 1864 this was confirmed by van Tieghem, and in 1874 by Cohn, who named the organism Micrococcus ureae. Pasteur and Cohn also pointed out that putrefaction is but a special case of fermentation, and before 1872 the doctrines of Pasteur were established with respect to Schizomycetes. Meanwhile two branches of inquiry had arisen, so to speak, from the above. In the first place, the ancient question of "spontaneous generation" received fresh impetus from the difficulty of keeping such minute organisms as bacteria from reaching and developing in organic infusions; and, secondly, the long-suspected analogies between the phenomena of fermentation and those of certain diseases again made themselves felt, as both became better understood. Needham in 1745 had declared that heated infusions of organic matter were not deprived of living beings; Spallanzani (1777) had replied that more careful heating and other precautions prevent the appearance of organisms in the fluid. Various experiments by Schwann, Helmholtz, Schultz, Schroeder, Dusch and others led to the refutation, step by step, of the belief that the more minute organisms, and particularly bacteria, arose de novo in the special cases quoted. Nevertheless, instances were adduced where the most careful heating of yolk of egg, milk, hay-infusions, &c., had failed,—the boiled infusions, &c., turning putrid and swarming with bacteria after a few hours.
In 1862 Pasteur repeated and extended such experiments, and paved the way for a complete explanation of the anomalies; Cohn in 1872 published confirmatory results; and it became clear that no putrefaction can take place without bacteria or some other living organism. In the hands of Brefeld, Burdon-Sanderson, de Bary, Tyndall, Roberts, Lister and others, the various links in the chain of evidence grew stronger and stronger, and every case adduced as one of "spontaneous generation" fell to the ground when examined. No case of so-called "spontaneous generation" has withstood rigid investigation; but the discussion contributed to more exact ideas as to the ubiquity, minuteness, and high powers of resistance to physical agents of the spores of Schizomycetes, and led to more exact ideas of antiseptic treatments. Methods were also improved, and the application of some of them to surgery at the hands of Lister, Koch and others has yielded results of the highest value.
Long before any clear ideas as to the relations of Schizomycetes to fermentation and disease were possible, various thinkers at different times had suggested that resemblances existed between the phenomena of certain diseases and those of fermentation, and the idea that a virus or contagium might be something of the nature of a minute organism capable of spreading and reproducing itself had been entertained. Such vague notions began to take more definite shape as the ferment theory of Cagniard de la Tour (1828), Schwann (1837) and Pasteur made way, especially in the hands of the last-named savant. From about 1870 onwards the "germ theory of disease" has passed into acceptance. P. F. O. Rayer in 1850 and Davaine had observed the bacilli in the blood of animals dead of anthrax (splenic fever), and Pollender discovered them anew in 1855. In 1863, imbued with ideas derived from Pasteur's researches on fermentation, Davaine reinvestigated the matter, and put forth the opinion that the anthrax bacilli caused the splenic fever; this was proved to result from inoculation. Koch in 1876 published his observations on Davaine's bacilli, placed beyond doubt their causal relation to splenic fever, discovered the spores and the saprophytic phase in the life-history of the organism, and cleared up important points in the whole question (figs. 7 and 9). In 1870 Pasteur had proved that a disease of silkworms was due to an organism of the nature of a bacterium; and in 1871 Oertel showed that a Micrococcus already known to exist in diphtheria is intimately concerned in producing that disease. In 1872, therefore, Cohn was already justified in grouping together a number of "pathogenous" Schizomycetes. Thus arose the foundations of the modern "germ theory of disease;" and, in the midst of the wildest conjectures and the worst of logic, a nucleus of facts was won, which has since grown, and is growing daily. Septicaemia, tuberculosis, glanders, fowl-cholera, relapsing fever, and other diseases are now brought definitely within the range of biology, and it is clear that all contagious and infectious diseases are due to the action of bacteria or, in a few cases, to fungi, or to protozoa or other animals.
[Illustration: FIG. 2.—The various phases of germination of spores of Bacillus ramosus (Fraenkel), as actually observed in hanging drops under very high powers.
A. The spore sown at 11 A.M., as shown at a, had swollen (b) perceptibly by noon, and had germinated by 3.30 P.M., as shown at c: in d at 6 P.M., and e at 8.30 P.M.; the resulting filament is segmenting into bacilli as it elongates, and at midnight (f) consisted of twelve such segments.
B, C. Similar series of phases in the order of the small letters in each case, and with the times of observation attached. At f and g occurs the breaking up of the filament into rodlets.
D. Germinating spores in various stages, more highly magnified, and showing the different ways of escape of the filament from the spore-membrane. (H. M. W.) ]
Other questions of the highest importance have arisen from the foregoing. About 1880 Pasteur first showed that Bacillus anthracis cultivated in chicken broth, with plenty of oxygen and at a temperature of 42-43deg C., lost its virulence after a few "generations," and ceased to kill even the mouse; Toussaint and Chauveau confirmed, and others have extended the observations. More remarkable still, animals inoculated with such "attenuated" bacilli proved to be curiously resistant to the deadly effects of subsequent inoculations of the non-attenuated form. In other words, animals vaccinated with the cultivated bacillus showed immunity from disease when reinoculated with the deadly wild form. The questions as to the causes and nature of the changes in the bacillus and in the host, as to the extent of immunity enjoyed by the latter, &c., are of the greatest interest and importance. These matters, however, and others such as phagocytosis (first described by Metchnikoff in 1884), and the epoch-making discovery of the opsonins of the blood by Wright, do not here concern us (see II. below).
[Sidenote: Form and Structure.]
MORPHOLOGY.—Sizes, Forms, Structure, &c.—The Schizomycetes consist of single cells, or of filamentous or other groups of cells, according as the divisions are completed at once or not. While some unicellular forms are less than 1 [micron] (.001 mm.) in diameter, others have cells measuring 4 [micron] or 5 [micron] or even 7 [micron] or 8 [micron], in thickness, while the length may vary from that of the diameter to many times that measurement. In the filamentous forms the individual cells are often difficult to observe until reagents are applied (e.g. fig. 14), and the length of the rows of cylindrical cells may be many hundred times greater than the breadth. Similarly, the diameters of flat or spheroidal colonies may vary from a few times to many hundred [Sidenote: Cell-wall.] times that of the individual cells, the divisions of which have produced the colony. The shape of the individual cell (fig. 1) varies from that of a minute sphere to that of a straight, curved, or twisted filament or cylinder, which is not necessarily of the same diameter throughout, and may have flattened, rounded, or even pointed ends. The rule is that the cells divide in one direction only—i.e. transverse to the long axis—and therefore produce aggregates of long cylindrical shape; but in rarer cases iso-diametric cells divide in two or three directions, producing flat, or spheroidal, or irregular colonies, the size of which is practically unlimited. The bacterial [v.03 p.0159] cell is always clothed by a definite cell-membrane, as was shown by the plasmolysing experiments of Fischer and others. Unlike the cell-wall of the higher plants, it gives usually no reactions of cellulose, nor is chitin present as in the fungi, but it consists of a proteid substance and is apparently a modification of the general protoplasm. In some cases, however, as in B. tuberculosis, analysis of the cell shows a large amount of cellulose. The cell-walls in some forms swell up into a gelatinous mass so that the cell appears to be surrounded in the unstained condition by a clear, transparent space. When the swollen wall is dense and regular in appearance the term "capsule" is applied to the sheath as in Leuconostoc. Secreted pigments (red, yellow, green and blue) are sometimes deposited in the wall, and some of the iron-bacteria have deposits of oxide of iron in the membranes.
[Illustration: FIG. 3.—Types of Zoogloea. (After Zopf.)
A. Mixed zoogloea found as a pellicle on the surface of vegetable infusions, &c.; it consists of various forms, and contains cocci (a) and rodlets, in series (b and c), &c.
B. Egg-shaped mass of zoogloea of Beggiatoa roseo-persicina (Bacterium rubescens of Lankester); the gelatinous swollen walls of the large crowded cocci are fused into a common gelatinous envelope.
C. Reticulate zoogloea of the same.
D, E, H. Colonies of Myconostoc enveloped in diffluent matrix.
F. Branched fruticose zoogloea of Cladothrix (slightly magnified).
G. Zoogloea of Bacterium merismopedioides, Zopf, containing cocci arranged in tablets.]
The substance of the bacterial cell when suitably prepared and stained shows in the larger forms a mass of homogeneous protoplasm containing irregular spaces, the vacuoles, which enclose a watery fluid. Scattered in the protoplasm arc usually one or more deeply-staining granules. The protoplasm itself may be tinged with colouring matter, bright red, yellow, &c., and may occasionally contain substances other than the deeply-staining granules. The occurrence of a starch-like substance which stains deep blue with iodine has been clearly shown in some forms even where the bacterium is growing on a medium containing no starch, as shown by Ward and others. In other forms a substance (probably glycogen or amylo-dextrin) which turns brown with iodine has been observed. Oil and fat drops have also been shown to occur, and in the sulphur-bacteria numerous fine granules of sulphur.
The question of the existence of a nucleus in the bacteria is one that has led to much discussion and is a problem of some difficulty. In the majority of forms it has not hitherto been possible to demonstrate a nucleus of the type which is so characteristic of the higher plants. Attention has accordingly been directed to the deeply-staining granules mentioned above, and the term chromatin-granules has been applied to them, and they have been considered to represent a rudimentary nucleus. That these granules consist of a material similar to the chromatin of the nucleus of higher forms is very doubtful, and the comparison with the nucleus of more highly organized cells rests on a very slender basis. The most recent works (Vejdovsky, Mencl), however, appear to show that nuclei of a structure and mode of division almost typical are to be found in some of the largest bacteria. It is possible that a similar structure has been overlooked or is invisible in other forms owing to their small size, and that there may be another type of nucleus—the diffuse nucleus—such as Schaudinn believed to be the case in B. butschlii. Many bacteria when suspended in a fluid exhibit a power of independent movement which is, of course, quite distinct from the Brownian movement—a non-vital phenomenon common to all finely-divided particles suspended in a fluid. Independent movement is effected by special motile organs, the cilia or flagella. These structures are invisible, with ordinary illumination in living cells or unstained preparations, and can only be made clearly visible by special methods of preparation and staining first used by Loeffler. By these methods the cilia are seen to be fine protoplasmic outgrowths of the cell (fig. 1) of the same nature as those of the zoospores and antherozoids of algae, mosses, &c. [Sidenote: Cilia.] These cilia appear to be attached to the cell-wall, being unaffected by plasmolysis, but Fischer states that they really are derived from the central protoplasm and pass through minute pores in the wall. The cilia may be present during a short period only in the life of a Schizomycete, and their number may vary according to the medium on which the organism is growing. Nevertheless, there is more or less constancy in the type of distribution, &c., of the cilia for each species when growing at its best. The chief results may be summed up as follows: some species, e.g. B. anthracis, have no cilia; others have only one flagellum at one pole (Monotrichous), e.g. Bacillus pyocyaneus (fig. 1, C, D), or one at each pole; others again have a tuft of several cilia [v.03 p.0160] at one pole (Lophotrichous), e.g. B. syncyaneus (fig. 1, E), or at each pole (Amphitrichous) (fig. 1, J, K, L); and, finally, many actively motile forms have the cilia springing all round (Peritrichous), e.g. B. vulgaris (fig. 1, G). It is found, however, that strict reliance cannot be placed on the distinction between the Monotrichous, Lophotrichous and Amphitrichous conditions, since one and the same species may have one, two or more cilia at one or both poles; nevertheless some stress may usually be laid on the existence of one or two as opposed to several—e.g. five or six or more—at one or each pole.
[Sidenote: Vegetative State.]
In Beggiatoa, a filamentous form, peculiar, slow, oscillatory movements are to be observed, reminding us of the movements of Oscillatoria among the Cyanophyceae. In these cases no cilia have been observed, and there is a firm cell-wall, so the movement remains quite unexplained.
[Illustration: FIG. 4.—Types of Spore-formation in Schizomycetes. (After Zopf.)
A. Various stages in the development of the endogenous spores in a Clostridium—the small letters indicate the order.
B. Endogenous spores of the hay bacillus.
C. A chain of cocci of Leuconostoc mesenterioides, with two "resting spores," i.e. arthrospores. (After van Tieghem.)
D. A motile rodlet with one cilium and with a spore formed inside.
E. Spore-formation in Vibrio-like (c) and Spirillum-like (a b, a) Schizomycetes.
F. Long rod-like form containing a spore (these are the so-called "Koepfchenbacterien" of German authors).
G. Vibrio form with spore. (After Prazmowski.)
H. Clostridium—one cell contains two spores. (After Prazmowski.)
I. Spirillum containing many spores (a), which are liberated at b by the breaking up of the parent cells.
K. Germination of the spore of the hay bacillus (B. subtilis)—the axis of growth of the germinal rodlet is at right angles to the long axis of the spore.
L. Germination of spore of Clostridium butyricum—the axis of growth coincides with the long axis of the spore.]
While many forms are fixed to the substratum, others are free, being in this condition either motile or immotile. The chief of these forms are described below.
Cocci: spherical or spheroidal cells, which, according to their relative (not very well defined) sizes are spoken of as Micrococci, Macrococci, and perhaps Monas forms.
Rods or rodlets: slightly or more considerably elongated cells which are cylindrical, biscuit-shaped or somewhat fusiform. The cylindrical forms are short, i.e. only three or four times as long as broad (Bacterium), or longer (Bacillus); the biscuit-shaped ones are Bacteria in the early stages of division. Clostridia, &c., are spindle-shaped.
Filaments really consist of elongated cylindrical cells which remain united end to end after division, and they may break up later into elements such as those described above. Such filaments are not always of the same diameter throughout, and their segmentation varies considerably. They may be free or attached at one (the "basal") end. A distinction is made between simple filaments (e.g. Leptothrix) and such as exhibit a false branching (e.g. Cladothrix).
Curved and spiral forms. Any of the elongated forms described above may be curved or sinuous or twisted into a corkscrew-like spiral instead of straight. If the sinuosity is slight we have the Vibrio form; if pronounced, and the spiral winding well marked, the forms are known as Spirillum, Spirochaete, &c. These and similar terms have been applied partly to individual cells, but more often to filaments consisting of several cells; and much confusion has arisen from the difficulty of defining the terms themselves.
In addition to the above, however, certain Schizomycetes present aggregates in the form of plates, or solid or hollow and irregular branched colonies. This may be due to the successive divisions occurring in two or three planes instead of only across the long axis (Sarcina), or to displacements of the cells after division.
Growth and Division.—Whatever the shape and size of the individual cell, cell-filament or cell-colony, the immediate visible results of active nutrition are elongation of the cell and its division into two equal halves, across the long axis, by the formation of a septum, which either splits at once or remains intact for a shorter or longer time. This process is then repeated and so on. In the first case the separated cells assume the character of the parent-cell whose division gave rise to them; in the second case they form filaments, or, if the further elongation and divisions of the cells proceed in different directions, plates or spheroidal or other shaped colonies. It not unfrequently happens, however, that groups of cells break away from their former connexion as longer or shorter straight or curved filaments, or as solid masses. In some filamentous forms this "fragmentation" into multicellular pieces of equal length or nearly so is a normal phenomenon, each partial filament repeating the growth, division and fragmentation as before (cf. figs. 2 and 6). By rapid division hundreds of thousands of cells may be produced in a few hours, and, according to the species and the conditions (the medium, temperature, &c.), enormous collections of isolated cells may cloud the fluid in which they are cultivated, or form deposits below or films on its surface; valuable characters are sometimes obtained from these appearances. When these dense "swarms" of vegetative cells become fixed in a matrix of their own swollen contiguous cell-walls, they pass over into a sort of resting state as a so-called zoogloea (fig. 3).
One of the most remarkable phenomena in the life-history of the Schizomycetes is the formation of this zoogloea stage, which corresponds to the "palmella" condition of the lower Algae. This occurs as a membrane on the surface of the medium, or as irregular clumps or branched masses (sometimes several inches across) submerged in it, and consists of more or less gelatinous matrix enclosing innumerable "cocci," "bacteria," or other elements of the Schizomycete concerned. Formerly regarded as a distinct genus—the natural fate of all the various [v.03 p.0161] forms—the zoogloea is now known to be a sort of resting condition of the Schizomycetes, the various elements being glued together, as it were, by their enormously swollen and diffluent cell-walls becoming contiguous. The zoogloea is formed by active division of single or of several mother-cells, and the progeny appear to go on secreting the cell-wall substance, which then absorbs many times its volume of water, and remains as a consistent matrix, in which the cells come to rest. The matrix—i.e. the swollen cell-walls—in some cases consists mainly of cellulose, in others chiefly of a proteid substance; the matrix in some cases is horny and resistant, in others more like a thick solution of gum. It is intelligible from the mode of formation that foreign bodies may become entangled in the gelatinous matrix, and compound zoogloeae may arise by the apposition of several distinct forms, a common event in macerating troughs (fig. 3, A). Characteristic forms may be assumed by the young zoogloea of different species,—spherical, ovoid, reticular, filamentous, fruiticose, lamellar, &c.,—but these vary considerably as the mass increases or comes in contact with others. Older zoogloeae may precipitate oxide of iron in the matrix, if that metal exists in small quantities in the medium. Under favourable conditions the elements in the zoogloea again become active, and move out of the matrix, distribute themselves in the surrounding medium, to grow and multiply as before. If the zoogloea is formed on a solid substratum it may become firm and horny; immersion in water softens it as described above.
; at 5.45, 20 [micron]; at 8 P.M., 70 [micron] and so on. Such curves show differences of steepness according to the temperature (see temp. curve), and to alterations of light (lamp) and darkness. (H. M. W.)]
[Sidenote: Measurement of growth.]
The growth of an ordinary bacterium consists in uniform elongation of the rodlet until its length is doubled, followed by division by a median septum, then by the simultaneous doubling in length of each daughter cell, again followed by the median division, and so on (figs. 13, 14). If the cells remain connected the resulting filament repeats these processes of elongation and subsequent division uniformly so long as the conditions are maintained, and very accurate measurements have been obtained on such a form, e.g. B. ramosus. If a rodlet in a hanging drop of nutrient gelatine is fixed under the microscope and kept at constant temperature, a curve of growth can be obtained recording the behaviour during many hours or days. The measured lengths are marked off on ordinates erected on an abscissa, along which the times are noted. The curve obtained on joining the former points then brings out a number of facts, foremost among which are (1) that as long as the conditions remain constant the doubling periods—i.e. the times taken by any portion of the filament to double its length—are constant, because each cell is equally active along the whole length; (2) there are optimum, minimum and maximum temperatures, other conditions remaining constant, at which growth begins, runs at its best and is soon exhausted, respectively; (3) that the most rapid cell-division and maximum growth do not necessarily accord with the best conditions for the life of the organism; and (4) that any sudden alteration of temperature brings about a check, though a slow rise may accelerate growth (fig. 8). It was also shown that exposure to light, dilution or exhaustion of the food-media, the presence of traces of poisons or metabolic products check growth or even bring it to a standstill; and the death or injury of any single cell in the filamentous series shows its effect on the curve by lengthening the doubling period, because its potential progeny have been put out of play. Hardy has shown that such a destruction of part of the filament may be effected by the attacks of another organism.
A very characteristic method of reproduction is that of spore-formation, and these minute reproductive bodies, which represent a resting stage of the organism, are now known in many forms. Formerly two kinds of spores were described, arthrospores and endospores. An arthrospore, however, is not a true spore but merely an ordinary vegetative cell which separates and passes into a condition of rest, and such may occur in forms which form endospores, e.g. B. subtilis, as well as in species not known to form endospores. The true spore or endospore begins with the appearance of a minute granule in the protoplasm of a vegetative cell; this granule enlarges and in a few hours has taken to itself all the protoplasm, secreted a thin but very resistive envelope, and is a ripe ovoid spore, smaller than the mother-cell and lying loosely in it (cf. figs. 6, 9, 10, and 11). In the case of the simplest and most minute Schizomycetes [v.03 p.0162] (Micrococcus, &c.) no definite spores have been discovered; any one of the vegetative micrococci may commence a new series of cell by growth and division. We may call these forms "asporous," at any rate provisionally.
The spore may be formed in short or long segments, the cell-wall of which may undergo change of form to accommodate itself to the contents. As a rule only one spore is formed in a cell, and the process usually takes place in a bacillar segment. In some cases the spore-forming protoplasm gives a blue reaction with iodine solutions. The spores may be developed in cells which are actively swarming, the movements not being interfered with by the process (fig. 4, D). The so-called "Koepfchenbacterien" of older writers are simply bacterioid segments with a spore at one end, the mother cell-wall having adapted itself to the outline of the spore (fig. 4, F). The ripe spores of Schizomycetes are spherical, ovoid or long-ovoid in shape and extremely minute (e.g. those of Bacillus subtilis measure 0.0012 mm. long by 0.0006 mm. broad according to Zopf), highly refractive and colourless (or very dark, probably owing to the high index of refraction and minute size). The membrane may be relatively thick, and even exhibit shells or strata.
The germination of the spores has now been observed in several forms with care. The spores are capable of germination at once, or they may be kept for months and even years, and are very resistant against desiccation, heat and cold, &c. In a suitable medium and at a proper temperature the germination is completed in a few hours. The spore swells and elongates and the contents grow forth to a cell like that which produced it, in some cases clearly breaking through the membrane, the remains of which may be seen attached to the young germinal rodlet (figs. 5, 9 and 11); in other cases the surrounding membrane of the spore swells and dissolves. The germinal cell then grows forth into the forms typical for the particular Schizomycete concerned.
The conditions for spore-formation differ. Anaerobic species usually require little oxygen, but aerobic species a free supply. Each species has an optimum temperature and many are known to require very special food-media. The systematic interference with these conditions has enabled bacteriologists to induce the development of so-called asporogenous races, in which the formation of spores is indefinitely postponed, changes in vigour, virulence and other properties being also involved, in some cases at any rate. The addition of minute traces of acids, poisons, &c., leads to this change in some forms; high temperature has also been used successfully.
The difficult subject of the classification of bacteria dates from the year 1872, when Cohn published his system, which was extended in 1875; this scheme has in fact dominated the study of bacteria ever since. Zopf in 1885 proposed a scheme based on the acceptance of extreme views of pleomorphism; his system, however, was extraordinarily impracticable and was recognized by him as provisional only. Systems have also been brought forward based on the formation of arthrospores and endospores, but as explained above this is eminently unsatisfactory, as arthrospores are not true spores and both kinds of reproductive bodies are found in one and the same form. Numerous attempts have been made to construct schemes of classification based on the power of growing colonies to liquefy gelatine, to secrete coloured pigments, to ferment certain media with evolution of carbon dioxide or other gases, or to induce pathological conditions in animals. None of these systems, which are chiefly due to the medical bacteriologists, has maintained its position, owing to the difficulty of applying the characters and to the fact that such properties are physiological and liable to great fluctuations in culture, because a given organism may vary greatly in such respects according to its degree of vitality at the time, its age, the mode of nutrition [v.03 p.0163] and the influence of external factors on its growth. Even when used in conjunction with purely morphological characters, these physiological properties are too variable to aid us in the discrimination of species and genera, and are apt to break down at critical periods. Among the more characteristic of these schemes adopted at various times may be mentioned those of Miquel (1891), Eisenberg (1891), and Lehmann and Neumann (1897). Although much progress has been made in determining the value and constancy of morphological characters, we are still in need of a sufficiently comprehensive and easily applied scheme of classification, partly owing to the existence in the literature of imperfectly described forms the life-history of which is not yet known, or the microscopic characters of which have not been examined with sufficient accuracy and thoroughness. [Sidenote: Fischer's Scheme.] The principal attempts at morphological classifications recently brought forward are those of de Toni and Trevisan (1889), Fischer (1897) and Migula (1897). Of these systems, which alone are available in any practical scheme of classification, the two most important and most modern are those of Fischer and Migula. The extended investigations of the former on the number and distribution of cilia (see fig. 1) led him to propose a scheme of classification based on these and other morphological characters, and differing essentially from any preceding one. This scheme may be tabulated as follows:—
I. ORDER—HAPLOBACTERINAE. Vegetative body unicellular; spheroidal, cylindrical or spirally twisted; isolated or connected in filamentous or other growth series.
1. Family—COCCACEAE. Vegetative cells spheroidal.
(a) Sub-family—ALLOCOCCACEAE. Division in all or any planes, colonies indefinite in shape and size, of cells in short chains, irregular clumps, pairs or isolated:— Micrococcus (Cohn), cells non-motile; Planococcus (Migula), cells motile.
(b) Sub-family—HOMOCOCCACEAE. Division planes regular and definite:—Sarcina (Goods.), cells non-motile; growth and division in three successive planes at right angles, resulting in packet-like groups; Planosarcina (Migula), as before, but motile; Pediococcus (Lindner), division planes at right angles in two successive planes, and cells in tablets of four or more; Streptococcus (Billr.), divisions in one plane only, resulting in chains of cells.
2. Family—BACILLACEAE. Vegetative cells cylindric (rodlets), ellipsoid or ovoid, and straight. Division planes always perpendicular to the long axis.
(a) Sub-family—BACILLEAE. Sporogenous rodlets cylindric, not altered in shape:—Bacillus (Cohn), non-motile; Bactrinium (Fischer), motile, with one polar flagellum (monotrichous); Bactrillum (Fischer), motile, with a terminal tuft of cilia (lophotrichous); Bactridium (Fischer), motile, with cilia all over the surface (peritrichous).
(b) Sub-family—CLOSTRIDIEAE. Sporogenous rodlets, spindle-shaped:—Clostridium (Prazm.), motile (peritrichous).
(c) Sub-family—PLECTRIDIEAE. Sporogenous rodlets, drumstick-shaped:—Plectridium (Fischer), motile (peritrichous).
3. Family—SPIRILLACEAE. Vegetative cells, cylindric but curved more or less spirally. Divisions perpendicular to the long axis:—Vibrio (Mueller-Loeffler), comma-shaped, motile, monotrichous; Spirillum (Ehrenb.), more strongly curved in open spirals, motile, lophotrichous; Spirochaete (Ehrenb.), spirally coiled in numerous close turns, motile, but apparently owing to flexile movements, as no cilia are found.
II. ORDER—TRICHOBACTERINAE. Vegetative body of branched or unbranched cell-filaments, the segments of which separate as swarm-cells (Gonidia).
1. Family—TRICHOBACTERIACEAE. Characters those of the Order.
(a) Filaments rigid, non-motile, sheathed:—Crenothrix (Cohn), filaments unbranched and devoid of sulphur particles; Thiothrix (Winogr.), as before, but with sulphur particles; Cladothrix (Cohn), filaments branched in a pseudo-dichotomous manner.
(b) Filaments showing slow pendulous and creeping movements, and with no distinct sheath:—Beggiatoa (Trev.), with sulphur particles.
The principal objections to this system are the following:—(1) The extraordinary difficulty in obtaining satisfactory preparations showing the cilia, and the discovery that these motile organs are not formed on all substrata, or are only developed during short periods of activity while the organism is young and vigorous, render this character almost nugatory. For instance, B. megatherium and B. subtilis pass in a few hours after commencement of growth from a motile stage with peritrichous cilia, into one of filamentous growth preceded by casting of the cilia. (2) By far the majority of the described species (over 1000) fall into the three genera—Micrococcus (about 400), Bacillus (about 200) and Bactridium (about 150), so that only a quarter or so of the forms are selected out by the other genera. (3) The monotrichous and lophotrichous conditions are by no means constant even in the motile stage; thus Pseudomonas rosea (Mig.) may have 1, 2 or 3 cilia at either end, and would be distributed by Fischer's classification between Bactrinium and Bactrillum, according to which state was observed. In Migula's scheme the attempt is made to avoid some of these difficulties, but others are introduced by his otherwise clever devices for dealing with these puzzling little organisms.
The question, What is an individual? has given rise to much difficulty, and around it many of the speculations regarding pleomorphism have centred without useful result. If a tree fall apart into its constituent cells periodically we should have the same difficulty on a larger and more complex scale. The fact that every bacterial cell in a species in most cases appears equally capable of performing all the physiological functions of the species has led most authorities, however, to regard it as the individual—a view which cannot be consistent in those cases where a simple or branched filamentous series exhibits differences between free apex and fixed base and so forth. It may be doubted whether the discussion is profitable, though it appears necessary in some cases—e.g. concerning pleomorphy—to adopt some definition of individual.
Myxobacteriaceae.—To the two divisions of bacteria, Haplobacterinae and Trichobacterinae, must now be added a third division, Myxobacterinae. One of the first members of this group, Chondromyces crocatus, was described as long ago as 1857 by Berkeley, but its nature was not understood and it was ascribed to the Hyphomycetes. In 1892, however, Thaxter rediscovered it and showed its bacterial nature, founding for it and some allied forms the group Myxobacteriaceae. Another form, which he described as Myxobacter, was shown later to be the same as Polyangium vitellinum described by Link in 1795, the exact nature of which had hitherto been in doubt. Thaxter's observations and conclusions were called in question by some botanists, but his later observations and those of Baur have established firmly the position of the group. The peculiarity of the group lies in the fact that the bacteria form plasmodium-like aggregations and build themselves up into sporogenous structures of definite form superficially similar to the cysts of the Mycetozoa (fig. 12). Most of the forms in question are found growing on the dung of herbivorous animals, but the bacteria occur not only in the alimentary canal of the animal but also free in the air. The Myxobacteria are most easily obtained by keeping at a temperature of 30-35deg C. in the dark dung which has lain exposed to the air for at least eight days. The high temperature is favourable to the growth of the bacteria but [v.03 p.0164] inimical to that of the fungi which are so common on this substratum.
[Sidenote: Function and life of bacteria.]
The discoveries that some species of nitrifying bacteria and perhaps pigmented forms are capable of carbon-assimilation, that others can fix free nitrogen and that a number of decompositions hitherto unsuspected are accomplished by Schizomycetes, have put the questions of nutrition and fermentation in quite new lights. Apart from numerous fermentation processes such as rotting, the soaking of skins for tanning, the preparation of indigo and of tobacco, hay, ensilage, &c., in all of which bacterial fermentations are concerned, attention may be especially directed to the following evidence of the supreme importance of Schizomycetes in agriculture and daily life. Indeed, nothing marks the attitude of modern bacteriology more clearly than the increasing attention which is being paid to useful fermentations. The vast majority of these organisms are not pathogenic, most are harmless and many are indispensable aids in natural operations important to man.
Fischer has proposed that the old division into saprophytes and parasites should be replaced by one which takes into account other peculiarities in the mode of nutrition of bacteria. The nitrifying, nitrogen-fixing, sulphur- and iron-bacteria he regards as monotrophic, i.e. as able to carry on one particular series of fermentations or decompositions only, and since they require no organic food materials, or at least are able to work up nitrogen or carbon from inorganic sources, he regards them as primitive forms in this respect and terms them Prototrophic. They may be looked upon as the nearest existing representatives of the primary forms of life which first obtained the power of working up non-living into living materials, and as playing a correspondingly important role in the evolution of life on our globe. The vast majority of bacteria, on the other hand, which are ordinarily termed saprophytes, are saprogenic, i.e. bring organic material to the putrefactive state—or saprophilous, i.e. live best in such putrefying materials—or become zymogenic, i.e. their metabolic products may induce blood-poisoning or other toxic effects (facultative parasites) though they are not true parasites. These forms are termed by Fischer Metatrophic, because they require various kinds of organic materials obtained from the dead remains of other organisms or from the surfaces of their bodies, and can utilize and decompose them in various ways (Polytrophic) or, if monotrophic, are at least unable to work them up. The true parasites—obligate parasites of de Bary—are placed by Fischer in a third biological group, Paratrophic bacteria, to mark the importance of their mode of life in the interior of living organisms where they live and multiply in the blood, juices or tissues.
[Sidenote: Nitrogen bacteria.]
When we reflect that some hundreds of thousands of tons of urea are daily deposited, which ordinary plants are unable to assimilate until considerable changes have been undergone, the question is of importance, What happens in the meantime? In effect the urea first becomes carbonate of ammonia by a simple hydrolysis brought about by bacteria, more and more definitely known since Pasteur, van Tieghem and Cohn first described them. Lea and Miquel further proved that the hydrolysis is due to an enzyme—urase—separable with difficulty from the bacteria concerned. Many forms in rivers, soil, manure heaps, &c., are capable of bringing about this change to ammonium carbonate, and much of the loss of volatile ammonia on farms is preventible if the facts are apprehended. The excreta of urea alone thus afford to the soil enormous stores of nitrogen combined in a form which can be rendered available by bacteria, and there are in addition the supplies brought down in rain from the atmosphere, and those due to other living debris. The researches of later years have demonstrated that a still more inexhaustible supply of nitrogen is made available by the nitrogen-fixing bacteria of the soil. There are in all cultivated soils forms of bacteria which are capable of forcing the inert free nitrogen to combine with other elements into compounds assimilable by plants. This was long asserted as probable before Winogradsky showed that the conclusions of M. P. E. Berthelot, A. Laurent and others were right, and that Clostridium pasteurianum, for instance, if protected from access of free oxygen by an envelope of aerobic bacteria or fungi, and provided with the carbohydrates and minerals necessary for its growth, fixes nitrogen in proportion to the amount of sugar consumed. This interesting case of symbiosis is equalled by yet another case. The work of numerous observers has shown that the free nitrogen of the atmosphere is brought into combination in the soil in the nodules filled with bacteria on the roots of Leguminosae, and since these nodules are the morphological expression of a symbiosis between the higher plant and the bacteria, there is evidently here a case similar to the last.
As regards the ammonium carbonate accumulating in the soil from the conversion of urea and other sources, we know from Winogradsky's researches that it undergoes oxidation in two stages owing to the activity of the so-called "nitrifying" bacteria (an unfortunate term inasmuch as "nitrification" refers merely to a particular phase of the cycle of changes undergone by nitrogen). It had long been known that under certain conditions large quantities of nitrate (saltpetre) are formed on exposed heaps of manure, &c., and it was supposed that direct oxidation of the ammonia, facilitated by the presence of porous bodies, brought this to pass. But research showed that this process of nitrification is dependent on temperature, aeration and moisture, as is life, and that while nitre-beds can infect one another, the process is stopped by sterilization. R. Warington, J. T. Schloessing, C. A. Muentz and others had proved that nitrification was promoted by some organism, when Winogradsky hit on the happy idea of isolating the organism by using gelatinous silica, and so avoiding the difficulties which Warington had shown to exist with the organism in presence of organic nitrogen, owing to its refusal to nitrify on gelatine or other nitrogenous media. Winogradsky's investigations resulted in the discovery that two kinds of bacteria are concerned in nitrification; one of these, which he terms the Nitroso-bacteria, is only capable of bringing about the oxidation of the ammonia to nitrous acid, and the astonishing result was obtained that [v.03 p.0165] this can be done, in the dark, by bacteria to which only pure mineral salts—e.g. carbonates, sulphates and chlorides of ammonium, sodium and magnesium—were added. In other words these bacteria can build up organic matter from purely mineral sources by assimilating carbon from carbon dioxide in the dark and by obtaining their nitrogen from ammonia. The energy liberated during the oxidation of the nitrogen is regarded as splitting the carbon dioxide molecule,—in green plants it is the energy of the solar rays which does this. Since the supply of free oxygen is dependent on the activity of green plants the process is indirectly dependent on energy derived from the sun, but it is none the less an astounding one and outside the limits of our previous generalizations. It has been suggested that urea is formed by polymerization of ammonium carbonate, and formic aldehyde is synthesized from CO2 and OH2. The Nitro-bacteria are smaller, finer and quite different from the nitroso-bacteria, and are incapable of attacking and utilizing ammonium carbonate. When the latter have oxidized ammonia to nitrite, however, the former step in and oxidize it still further to nitric acid. It is probable that important consequences of these actions result from the presence of nitrifying bacteria in rotten stone, decaying bricks, &c., where all the conditions are realized for preparing primitive soil, the breaking up of the mineral constituents being a secondary matter. That "soil" is thus prepared on barren rocks and mountain peaks may be concluded with some certainty.
In addition to the bacterial actions which result in the oxidization of ammonia to nitrous acid, and of the latter to nitric acid, the reversal of such processes is also brought about by numerous bacteria in the soil, rivers, &c. Warington showed some time ago that many species are able to reduce nitrates to nitrites, and such reduction is now known to occur very widely in nature. The researches of Gayon and Dupetit, Giltay and Aberson and others have shown, moreover, that bacteria exist which carry such reduction still further, so that ammonia or even free nitrogen may escape. The importance of these results is evident in explaining an old puzzle in agriculture, viz. that it is a wasteful process to put nitrates and manure together on the land. Fresh manure abounds in de-nitrifying bacteria, and these organisms not only reduce the nitrates to nitrites, even setting free nitrogen and ammonia, but their effect extends to the undoing of the work of what nitrifying bacteria may be present also, with great loss. The combined nitrogen of dead organisms, broken down to ammonia by putrefactive bacteria, the ammonia of urea and the results of the fixation of free nitrogen, together with traces of nitrogen salts due to meteoric activity, are thus seen to undergo various vicissitudes in the soil, rivers and surface of the globe generally. The ammonia may be oxidized to nitrites and nitrates, and then pass into the higher plants and be worked up into proteids, and so be handed on to animals, eventually to be broken down by bacterial action again to ammonia; or the nitrates may be degraded to nitrites and even to free nitrogen or ammonia, which escapes.
[Sidenote: Bacteria and Leguminosae.]
That the Leguminosae (a group of plants including peas, beans, vetches, lupins, &c.) play a special part in agriculture was known even to the ancients and was mentioned by Pliny (Historia Naturalis, viii). These plants will not only grow on poor sandy soil without any addition of nitrogenous manure, but they actually enrich the soil on which they are grown. Hence leguminous plants are essential in all rotation of crops. By analysis it was shown by Schulz-Lupitz in 1881 that the way in which these plants enrich the soil is by increasing the nitrogen-content. Soil which had been cultivated for many years as pasture was sown with lupins for fifteen years in succession; an analysis then showed that the soil contained more than three times as much nitrogen as at the beginning of the experiment. The only possible source for this increase was the atmospheric nitrogen. It had been, however, an axiom with botanists that the green plants were unable to use the nitrogen of the air. The apparent contradiction was explained by the experiments of H. Hellriegel and Wilfarth in 1888. They showed that, when grown on sterilized sand with the addition of mineral salts, the Leguminosae were no more able to use the atmospheric nitrogen than other plants such as oats and barley. Both kinds of plants required the addition of nitrates to the soil. But if a little water in which arable soil had been shaken up was added to the sand, then the leguminous plants flourished in the absence of nitrates and showed an increase in nitrogenous material. They had clearly made use of the nitrogen of the air. When these plants were examined they had small swellings or nodules on their roots, while those grown in sterile sand without soil-extract had no nodules. Now these peculiar nodules are a normal characteristic of the roots of leguminous plants grown in ordinary soil. The experiments above mentioned made clear for the first time the nature and activity of these nodules. They are clearly the result of infection (if the soil extract was boiled before addition to the sand no nodules were produced), and their presence enabled the plant to absorb the free nitrogen of the air.
The work of recent investigators has made clear the whole process. In ordinary arable soil there exist motile rod-like bacteria, Bacterium radicicola. These enter the root-hairs of leguminous plants, and passing down the hair in the form of a long, slimy (zoogloea) thread, penetrate the tissues of the root. As a result the tissues become hypertrophied, producing the well-known nodule. In the cells of the nodule the bacteria multiply and develop, drawing material from their host. Many of the bacteria exhibit curious involution forms ("bacteroids"), which are finally broken down and their products absorbed by the plant. The nitrogen of the air is absorbed by the nodules, being built up into the bacterial cell and later handed on to the host-plant. It appears from the observations of Maze that the bacterium can even absorb free nitrogen when grown in cultures [v.03 p.0166] outside the plant. We have here a very interesting case of symbiosis as mentioned above. The green plant, however, always keeps the upper hand, restricting the development of the bacteria to the nodules and later absorbing them for its own use. It should be mentioned that different genera require different races of the bacterium for the production of nodules.
The important part that these bacteria play in agriculture led to the introduction in Germany of a commercial product (the so-called "nitragin") consisting of a pure culture of the bacteria, which is to be sprayed over the soil or applied to the seeds before sowing. This material was found at first to have a very uncertain effect, but later experiments in America, and the use of a modified preparation in England, under the direction of Professor Bottomley, have had successful results; it is possible that in the future a preparation of this sort will be widely used.
The apparent specialization of these bacteria to the leguminous plants has always been a very striking fact, for similar bacterial nodules are known only in two or three cases outside this particular group. However, Professor Bottomley announced at the meeting of the British Association for the Advancement of Science in 1907 that he had succeeded in breaking down this specialization and by a suitable treatment had caused bacteria from leguminous nodules to infect other plants such as cereals, tomato, rose, with a marked effect on their growth. If these results are confirmed and the treatment can be worked commercially, the importance to agriculture of the discovery cannot be overestimated; each plant will provide, like the bean and vetch, its own nitrogenous manure, and larger crops will be produced at a decreased cost.
Another important advance is in our knowledge of the part played by bacteria in the circulation of carbon in nature. The enormous masses of cellulose deposited annually on the earth's surface are, as we know, principally the result of chlorophyll action on the carbon dioxide of the atmosphere decomposed by energy derived from the sun; and although we know little as yet concerning the magnitude of other processes of carbon-assimilation—e.g. by nitrifying bacteria—it is probably comparatively small. Such cellulose is gradually reconverted into water and carbon dioxide, but for some time nothing positive was known as to the agents which thus break up the paper, rags, straw, leaves and wood, &c., accumulating in cesspools, forests, marshes and elsewhere in such abundance. The work of van Tieghem, van Senus, Fribes, Omeliansky and others has now shown that while certain anaerobic bacteria decompose the substance of the middle lamella—chiefly pectin compounds—and thus bring about the isolation of the cellulose fibres when, for instance, flax is steeped or "retted," they are unable to attack the cellulose itself. There exist in the mud of marshes, rivers and cloacae, &c., however, other anaerobic bacteria which decompose cellulose, probably hydrolysing it first and then splitting the products into carbon dioxide and marsh gas. When calcium sulphate is present, the nascent methane induces the formation of calcium carbonate, sulphuretted hydrogen and water. We have thus an explanation of the occurrence of marsh gas and sulphuretted hydrogen in bogs, and it is highly probable that the existence of these gases in the intestines of herbivorous animals is due to similar putrefactive changes in the undigested cellulose remains.
[Sidenote: Sulphur bacteria.]
Cohn long ago showed that certain glistening particles observed in the cells of Beggiatoa consist of sulphur, and Winogradsky and Beyerinck have shown that a whole series of sulphur bacteria of the genera Thiothrix, Chromatium, Spirillum, Monas, &c., exist, and play important parts in the circulation of this element in nature, e.g. in marshes, estuaries, sulphur springs, &c. When cellulose bacteria set free marsh gas, the nascent gas reduces sulphates—e.g. gypsum—with liberation of SH2, and it is found that the sulphur bacteria thrive under such conditions by oxidizing the SH2 and storing the sulphur in their own protoplasm. If the SH2 runs short they oxidize the sulphur again to sulphuric acid, which combines with any calcium carbonate present and forms sulphate again. Similarly nascent methane may reduce iron salts, and the black mud in which these bacteria often occur owes its colour to the FeS formed. Beyerinck and Jegunow have shown that some partially anaerobic sulphur bacteria can only exist in strata at a certain depth below the level of quiet waters where SH2 is being set free below by the bacterial decompositions of vegetable mud and rises to meet the atmospheric oxygen coming down from above, and that this zone of physiological activity rises and falls with the variations of partial pressure of the gases due to the rate of evolution of the SH2. In the deeper parts of this zone the bacteria absorb the SH2, and, as they rise, oxidize it and store up the sulphur; then ascending into planes more highly oxygenated, oxidize the sulphur to SO3. These bacteria therefore employ SH2 as their respiratory substance, much as higher plants employ carbohydrates—instead of liberating energy as heat by the respiratory combustion of sugars, they do it by oxidizing hydrogen sulphide. Beyerinck has shown that Spirillum desulphuricans, a definite anaerobic form, attacks and reduces sulphates, thus undoing the work of the sulphur bacteria as certain de-nitrifying bacteria reverse the operations of nitro-bacteria. Here again, therefore, we have sulphur, taken [v.03 p.0167] into the higher plants as sulphates, built up into proteids, decomposed by putrefactive bacteria and yielding SH2 which the sulphur bacteria oxidize, the resulting sulphur is then again oxidized to SO3 and again combined with calcium to gypsum, the cycle being thus complete.
[Sidenote: Iron bacteria.]
Chalybeate waters, pools in marshes near ironstone, &c, abound in bacteria, some of which belong to the remarkable genera Crenothrix, Cladothrix and Leptothrix, and contain ferric oxide, i.e. rust, in their cell-walls. This iron deposit is not merely mechanical but is due to the physiological activity of the organism which, according to Winogradsky, liberates energy by oxidizing ferrous and ferric oxide in its protoplasm—a view not accepted by H. Molisch. The iron must be in certain soluble conditions, however, and the soluble bicarbonate of the protoxide of chalybeate springs seems most favourable, the hydrocarbonate absorbed by the cells is oxidized, probably thus—
2FeCO_3 + 3OH_2 + O = Fe_2(OH)_6 + 2CO_2.
The ferric hydroxide accumulates in the sheath, and gradually passes into the more insoluble ferric oxide. These actions are of extreme importance in nature, as their continuation results in the enormous deposits of bog-iron ore, ochre, and—since Molisch has shown that the iron can be replaced by manganese in some bacteria—of manganese ores.
[Sidenote: Pigment bacteria.]
Considerable advances in our knowledge of the various chromogenic bacteria have been made by the studies of Beyerinck, Lankester, Engelmann, Ewart and others, and have assumed exceptional importance owing to the discovery that Bacteriopurpurin—the red colouring matter contained in certain sulphur bacteria—absorbs certain rays of solar energy, and enables the organism to utilize the energy for its own life-purposes. Engelmann showed, for instance, that these red-purple bacteria collect in the ultra-red, and to a less extent in the orange and green, in bands which agree with the absorption spectrum of the extracted colouring matter. Not only so, but the evident parallelism between this absorption of light and that by the chlorophyll of green plants, is completed by the demonstration that oxygen is set free by these bacteria—i.e. by means of radiant energy trapped by their colour-screens the living cells are in both cases enabled to do work, such as the reduction of highly oxidized compounds.
The most recent observations of Molisch seem to show that bacteria possessing bacteriopurpurin exhibit a new type of assimilation—the assimilation of organic material under the influence of light. In the case of these red-purple bacteria the colouring matter is contained in the protoplasm of the cell, but in most chromogenic bacteria it occurs as excreted pigment on and between the cells, or is formed by their action in the medium. Ewart has confirmed the principal conclusions concerning these purple, and also the so-called chlorophyll bacteria (B. viride, B. chlorinum, &c.), the results going to show that these are, as many authorities have held, merely minute algae. The pigment itself may be soluble in water, as is the case with the blue-green fluorescent body formed by B. pyocyaneus, B. fluorescens and a whole group of fluorescent bacteria. Neelson found that the pigment of B. cyanogenus gives a band in the yellow and strong lines at E and F in the solar spectrum—an absorption spectrum almost identical with that of triphenyl-rosaniline. In the case of the scarlet and crimson red pigments of B. prodigiosus, B. ruber, &c., the violet of B. violacens, B janthinus, &c., the red-purple of the sulphur bacteria, and indeed most bacterial pigments, solution in water does not occur, though alcohol extracts the colour readily. Finally, there are a few forms which yield their colour to neither alcohol nor water, e.g. the yellow Micrococcus cereus flavus and the B. berolinensis. Much work is still necessary before we can estimate the importance of these pigments. Their spectra are only imperfectly known in a few cases, and the bearing of the absorption on the life-history is still a mystery. In many cases the colour-production is dependent on certain definite conditions—temperature, presence of oxygen, nature of the food-medium, &c. Ewart's important discovery that some of these lipochrome pigments occlude oxygen, while others do not, may have bearings on the facultative anaerobism of these organisms.
[Sidenote: Dairy bacteria.]
A branch of bacteriology which offers numerous problems of importance is that which deals with the organisms so common in milk, butter and cheese. Milk is a medium not only admirably suited to the growth of bacteria, but, as a matter of fact, always contaminated with these organisms in the ordinary course of supply. F. Lafar has stated that 20% of the cows in Germany suffer from tuberculosis, which also affected 17.7% of the cattle slaughtered in Copenhagen between 1891 and 1893, and that one in every thirteen samples of milk examined in Paris, and one in every nineteen in Washington, contained tubercle bacilli. Hence the desirability of sterilizing milk used for domestic purposes becomes imperative.
[Illustration: FIG. 18.—A similar preparation to fig. 17, except that two slit-like openings of equal length allowed the light to pass, and that the light was that of the electric arc passed through a quartz prism and casting a powerful spectrum on the plate. The upper slit was covered with glass, the lower with quartz. The bacteria were killed over the clear areas shown. The left-hand boundary of the clear area corresponds to the line F (green end of the blue), and the beginning of the ultra-violet was at the extreme right of the upper (short) area. The lower area of bactericidal action extends much farther to the right, because the quartz allows more ultra-violet rays to pass than does glass. The red-yellow-green to the left of F were without effect. (H. M. W.) ]
No milk is free from bacteria, because the external orifices of the milk-ducts always contain them, but the forms present in the normal fluid are principally those which induce such changes as the souring or "turning" so frequently observed in standing milk (these were examined by Lord Lister as long ago as 1873-1877, though several other species are now known), and those which bring about the various changes and fermentations in butter and cheese made from it. The presence of foreign germs, which may gain the upper hand and totally destroy the flavours of butter and cheese, has led to the search for those particular forms to which the approved properties are due. A definite bacillus to which the peculiarly fine flavour of certain butters is due, is said to be largely employed in pure cultures in American dairies, and in Denmark certain butters are said to keep fresh much longer owing to the use of pure cultures and the treatment employed to suppress the forms which cause rancidity. Quite distinct is the search for the germs which cause undesirable changes, or "diseases"; and great strides have been made in discovering the bacteria concerned in rendering milk "ropy," butter "oily" and "rancid," &c. Cheese in its numerous forms contains myriads of bacteria, and some of these are now known to be concerned in the various processes of ripening and other changes affecting the product, and although little is known as to the exact part played by any species, practical applications of the discoveries of the decade 1890-1900 have been made, e.g. Edam cheese. The Japanese have cheeses resulting from the bacterial fermentation of boiled Soja beans.
[Sidenote: Thermophilous bacteria.]
That bacterial fermentations are accompanied by the evolution of heat is an old experience; but the discovery that the "spontaneous" combustion of sterilized cotton-waste does not occur simply if moist and freely exposed to oxygen, but results when the washings of fresh waste are added, has led to clearer proof that the heating of hay-stacks, hops, tobacco and other vegetable products is due to the vital activity of bacteria and fungi, and is physiologically a consequence of respiratory processes like those in malting. It seems fairly established that when the preliminary heating process of fermentation is drawing to a close, the cotton, hay, &c., having been converted into a highly porous friable and combustible mass, may then ignite in certain circumstances by the occlusion of oxygen, just as ignition is induced by finely divided metals. A remarkable point in this connexion has always been the necessary conclusion that the living bacteria concerned must be exposed to temperatures of at least 70deg C. in the hot heaps. Apart from the resolution of doubts as to the power of spores to withstand such temperatures for long periods, the discoveries of Miquel, Globig and others have shown that there are numerous bacteria which will grow and divide at such temperatures, e.g. B. thermophilus, from sewage, which is quite active at 70deg C., and B. Ludwigi and B. ilidzensis, &c., from hot springs, &c.
[Sidenote: Phosphorescent bacteria.]
The bodies of sea fish, e.g. mackerel and other animals, have long been known to exhibit phosphorescence. This phenomenon is due to the activity of a whole series of marine bacteria of various genera, the examination and cultivation of which have been successfully carried out by Cohn, Beyerinck, Fischer and others. The cause of the phosphorescence is still a mystery. The suggestion that it is due to the oxidation of a body excreted by the bacteria seems answered by the failure to filter off or extract any such body. Beyerinck's view that it occurs at the moment peptones are worked up into the protoplasm cannot be regarded as proved, and the same must be said of the suggestion that the phosphorescence is due to the oxidation of phosphoretted hydrogen. The conditions of phosphorescence are, the presence of free oxygen, and, generally, a relatively low temperature, together with a medium containing sodium chloride, and peptones, but little or no carbohydrates. Considerable differences occur in these latter respects, however, and interesting results were obtained by Beyerinck with mixtures of species possessing different powers of enzyme action as regards carbohydrates. Thus, a form termed Photobacterium phosphorescens by Beyerinck will absorb maltose, and will become luminous if that sugar is present, whereas P. Pflugeri is indifferent to maltose. If then we prepare densely inseminated plates of these two bacteria in gelatine food-medium to which starch is added as the only carbohydrate, the bacteria grow but do not phosphoresce. If we now streak these plates with an organism, e.g. a yeast, which saccharifies starch, it is possible to tell whether maltose or levulose and fructose are formed; if the former, only those plates containing P. phosphorescens will become luminous; if the latter, only those containing P. Pflugeri. The more recent researches of Molisch have shown that the luminosity of ordinary butcher's meat under appropriate conditions is quite a common occurrence. Thus of samples of meat bought in Prague and kept in a cool room for about two days, luminosity was present in 52% of the samples in the case of beef, 50% for veal, and 39% for liver. If the meat was treated previously with a 3% salt solution, 89% of the samples of beef and 65% of the samples of horseflesh were found to exhibit this phenomenon. The cause of this luminosity is Micrococcus phosphorens, an immotile round, or almost round organism. This organism is quite distinct from that causing the luminosity of marine fish.
[Sidenote: Oxidizing bacteria.]
It has long been known that the production of vinegar depends on the oxidization of the alcohol in wine or beer to acetic acid, the chemical process being probably carried out in two stages, viz. the oxidation of the alcohol leading to the formation of aldehyde and water, and the further oxidation of the aldehyde to acetic acid. The process may even go farther, and the acetic acid be oxidized to CO2 and OH2; the art of the vinegar-maker is directed to preventing the accomplishment of the last stage. These oxidations are brought about by the vital activity of several bacteria, of which four—Bacterium aceti, B. pasteurianum, B. kutzingianum, and B. xylinum—have been thoroughly studied by Hansen and A. Brown. It is these bacteria which form the zoogloea of the "mother of vinegar," though this film may contain other organisms as well. The idea that this film of bacteria oxidizes the alcohol beneath by merely condensing atmospheric oxygen in its interstices, after the manner of spongy platinum, has long been given up; but the explanation of the action as an incomplete combustion, depending on the peculiar respiration of these organisms—much as in the case of nitrifying and sulphur bacteria—is not clear, though the discovery that the acetic bacteria will not only oxidize alcohol to acetic acid, but further oxidize the latter to CO2 and OH2 supports the view that the alcohol is absorbed by the organism and employed as its respirable substance. Promise of more light on these oxidation fermentations is afforded by the recent discovery that not only bacteria and fungi, but even the living cells of higher plants, contain peculiar enzymes which possess the remarkable property of "carrying" oxygen—much as it is carried in the sulphuric acid chamber—and which have therefore been termed oxydases. It is apparently the presence of these oxydases which causes certain wines to change colour and alter in taste when poured from bottle to glass, and so exposed to air.
[Sidenote: Bacteria and light.]
Much as the decade from 1880 to 1890 abounded with investigations on the reactions of bacteria to heat, so the following decade was remarkable for discoveries regarding the effects of other forms of radiant energy. The observations of Downes and Blunt in 1877 left it uncertain whether the bactericidal effects in broth cultures exposed to solar rays were due to thermal action or not. Further investigations, in which Arloing, Buchner, Chmelewski, and others took part, have led to the proof that rays of light alone are quite capable of killing these organisms. The principal questions were satisfactorily settled by Marshall Ward's experiments in 1892-1893, when he showed that even the spores of B. anthracis, which withstand temperatures of 100deg C. and upwards, can be killed by exposure to rays of reflected light at temperatures far below anything injurious, or even favourable to growth. He also showed that the bactericidal action takes place in the absence of food materials, thus proving that it is not merely a poisoning effect of the altered medium. The principal experiments also indicate that it is the rays of highest refrangibility—the blue-violet and ultra-violet rays of the spectrum—which bring about the destruction of the organisms (figs. 17, 18). The practical effect of the bactericidal action of solar light is the destruction of enormous quantities of germs in rivers, the atmosphere and other exposed situations, and experiments have shown that it is especially the pathogenic bacteria—anthrax, typhoid, &c.—which thus succumb to light-action; the discovery that the electric arc is very rich in bactericidal rays led to the hope that it could be used for disinfecting purposes in hospitals, but mechanical difficulties intervene. The recent application of the action of bactericidal rays to the cure of lupus is, however, an extension of the same discovery. Even when the light is not sufficiently intense, or the exposure is too short to kill the spores, the experiments show that attenuation of virulence [v.03 p.0169] may result, a point of extreme importance in connexion with the lighting and ventilation of dwellings, the purification of rivers and streams, and the general diminution of epidemics in nature.
[Sidenote: Bacteria and cold.]
As we have seen, thermophilous bacteria can grow at high temperatures, and it has long been known that some forms develop on ice. The somewhat different question of the resistance of ripe spores or cells to extremes of heat and cold has received attention. Ravenel, Macfadyen and Rowland have shown that several bacilli will bear exposure for seven days to the temperature of liquid air (-192deg C. to -183deg C.) and again grow when put into normal conditions. More recent experiments have shown that even ten hours' exposure to the temperature of liquid hydrogen -252deg C. (21deg on the absolute scale) failed to kill them. It is probable that all these cases of resistance of seeds, spores, &c., are to be connected with the fact that completely dry albumin does not lose its coagulability on heating to 110deg C. for some hours, since it is well known that completely ripe spores and dry heat are the conditions of extreme experiments.