The modes of origin of lowest organisms / including a discussion of the experiments of M. Pasteur

by H. Charlton Bastian

If germs derived from the air are the sole causes of putrefaction, then, surely, deprivation from air ought to be just as efficacious as any process of filtration of air—more especially when the filtration or the deprivation have a common starting point. And the mode of procedure, in both cases, is precisely the same up to a certain point. A fluid is boiled for a short time in order to kill the germs which may be within the flask, and to expel its previously contained air. At a certain stage of the ebullition, this may be arrested, if we have to do with a bent-neck flask, or one whose neck is plugged with cotton-wool, and no change, it is said, will subsequently take place in the contained fluid, because the air which enters is, by either of these means, filtered from its germs. But if, whilst ebullition continued, the neck of the flask had been hermetically sealed—so as altogether to prevent the re-ingress of air—and if the fluid, thus contained in vacuo, would nevertheless undergo fermentation, obviously the former explanation must be altogether shelved.

In the face of M. Pasteur’s explanations, and those of Professor Huxley, these frequent positive results with fluids contained in vacuo are absolutely contradictory. There may naturally arise, therefore, a very grave doubt as to the validity of the explanation adduced by M. Pasteur, and adopted by Professor Huxley and others.

All these experiments to which I have been alluding are based upon the supposition (assented to by Pasteur and Huxley) that Bacteria which pre-existed in the solution would certainly be destroyed by its being raised for a few minutes to a temperature of 212° F. This conclusion is, I believe, perfectly correct,[26] and in support thereof I will adduce the following additional information.

Limits of ‘Vital Resistance’ to Heat displayed by Bacteria and Torulæ.

After stating elsewhere[27], that Vibriones are partly broken up or disintegrated by an exposure for a few minutes to a temperature of 212° F. in an infusion which is being boiled, and also that, in all probability, the life of Bacteria would be destroyed by such a treatment, I made the following remarks:—“With reference to these organisms, however, one caution is necessary to be borne in mind by the experimenter. The movements of monads and Bacteria may be, and frequently are, of two kinds. The one variety does not differ in the least from the mere molecular or Brownian movement, which may be witnessed in similarly minute, not-living particles immersed in fluids. Whilst the other seems to be purely vital—that is, dependent upon their properties as living things. These vital movements are altogether different from the mere dancing oscillations which not-living particles display, as may be seen when the monad or Bacterium darts about over comparatively large areas, so as frequently to disappear from the field. After an infusion has been exposed for a second or two to the boiling temperature, these vital movements no longer occur, though almost all the monads and Bacteria may be seen to display the Brownian movement in a well-marked degree. They seem to be reduced by the shortest exposure to a temperature of 100° C. to the condition of mere not-living particles, and then they become subjected to the unimpaired influence of the physical conditions which determine these movements.” I now have various facts to add in confirmation of these conclusions, and in extension of our knowledge concerning the vital resistance to heat of Bacteria and Torulæ.

It would be a most important step if we could ascertain some means by which these primary movements of living Bacteria might be distinguished from the secondary, or communicated, movements of not-living particles. In many cases, organisms that are truly living may only exhibit very languid movements, which, as movements, are quite indistinguishable from those that the same Bacteria may display when they are really dead. Because the movements, therefore, are of this doubtful character, persons are apt, unfairly, to argue that the Bacteria which present them, are no more living than are the minute particles of carbon obtained from the flame of a lamp, which may exhibit similar movements. This, however, is a point of view which becomes obviously misleading if too much stress is laid upon it; and it is more especially so in this case, when those Bacteria which display the most characteristic sign of vitality—viz., “spontaneous” division or reproduction—do, at the time, almost always exhibit only the same languid movements. Mobility is, in fact, not an essential characteristic of living Bacteria, whilst the occurrence of the act of reproduction is the most indubitable sign of their life. It should be remembered, therefore, that any Bacteria which are almost motionless, or which exhibit mere Brownian movements, may be living, whilst those which spontaneously divide and reproduce, are certainly alive—whatever may be the kind of movement they present.

In any particular case, however, can we decide whether Bacteria, that have been submitted to a given temperature, and which exhibit movements resembling those known as Brownian, are really dead or living? If the movements are primary, or dependent upon the inherent molecular activity of the organism itself, they ought, it might be argued, to continue when the molecules of the fluid are at rest; if, on the other hand, they are mere secondary or communicated movements, impressed upon the organisms as they would be upon any other similarly minute particles, by the molecular oscillations of the fluid in which they are contained, then the movements ought to grow less, and gradually cease, as the fluid approaches a state of molecular rest—if this be attainable. Following out this idea, some months ago, I first tested the correctness of the assumption by experimenting with fluids containing various kinds of not-living particles; such as carbon-particles from the flame of a lamp, or freshly precipitated baric sulphate. However perfect may have been the Brownian movements when portions of these fluids were first examined beneath a covering-glass, they always gradually diminished, after the specimen had been mounted by surrounding the covering-glass with some cement or varnish. Thus prepared, no evaporation could take place from the thin film of fluid, and after one, three, four, or more hours—the slide remaining undisturbed—most of the particles had subsided, and were found to have come to a state of rest. In order still further to test these views, I took an infusion of turnip, containing a multitude of Bacteria whose movements were of the languid description, and divided it into two portions. One of these portions was boiled for about a minute, whilst the other was not interfered with. Then, after the boiled solution had been cooled, a drop was taken from each and placed at some little distance from one another on the same glass slip; covering-glasses half an inch in diameter were laid on, and the superfluous fluid beneath each was removed by a piece of blotting-paper. When only the thinnest film of fluid was left, the covering-glasses were surrounded by a thick, quickly-drying cement.[28] Examined with the microscope immediately afterwards, it was generally found that the Bacteria which had been boiled presented a shrunken and shrivelled aspect—whilst some of them were more or less disintegrated—though, as far as movement was concerned, there was little to distinguish that which they manifested, from that of their plumper-looking relatives which had not been boiled.

If the specimens were examined again after twenty-four or more hours, there was still very little difference perceptible between them, as regards their movements. And the same was the case when the specimens were examined after a lapse of some days or weeks. One important difference does, however, soon become obvious. The Bacteria which have not been boiled, undergo a most unmistakeable increase within their imprisoned habitat; whilst those which have been boiled, do not increase. The two films may be almost colourless at first (if the Bacteria are not very abundant), but after a few days, that composed of unboiled fluid begins to show an obvious and increasing cloudiness, which is never manifested by the other. Microscopical examination shows that this cloudiness is due to a proportionate increase in the number of Bacteria.

Is the continuance of the movements of the organisms which had been boiled attributable to their extreme lightness, and to the slight difference between their specific gravity and that of the fluid in which they are immersed? I soon became convinced that this was one, if not the chief reason, when I found that Bacteria which had been submitted to very much higher temperatures, behaved in precisely the same manner as those which had been merely boiled, and also that other particles which—though obviously dead—had a similar specific lightness, also continued to exhibit their Brownian movements for days and weeks. This was the case more especially with the minute fat particles in a mounted specimen of boiled milk,[29] and also with very minute particles which were gradually precipitated[30] from a hay infusion that had been heated to 302° F. for four hours. Trials with many different substances, indeed, after a time convinced me that the most rapid cessation of Brownian movements in stationary films,[31] occurred where the particles were heavy or large; and that the duration of the movement was more and more prolonged, as the particles experimented with, were lighter or more minute. So that, when we have to do with Bacteria, the minute oil globules of milk, or with other similarly light particles, the movements continue for an indefinite time, and are, in part, mere exponents of the molecular unrest of the fluid. They are always capable of being increased or renewed by the incidence of heat or other disturbing agencies.