Part 63

With such chambers I tested, in the autumn and winter of 1875-6, infusions of the most various kinds, embracing natural animal liquids, the flesh and viscera of domestic animals, game, fish, and vegetables. More than fifty chambers, each with its series of infusions, were tested, many of them repeatedly. There was no shade of uncertainty in any of the results. In every instance we had, within the chamber, perfect limpidity and sweetness, which in some cases lasted for more than a year--without the chamber, with the same infusion, putridity and its characteristic smells. In no instance was the least countenance lent to the notion that an infusion deprived by heat of its inherent life, and placed in contact with air cleansed of its visibly suspended matter, has any power to generate life anew.

Remembering then the number and variety of the infusions employed, and the strictness of our adherence to the rules of preparation laid down by the heterogenists themselves; remembering that we have operated upon the very substances recommended by them as capable of furnishing, even in untrained hands, easy and decisive proofs of spontaneous generation, and that we have added to their substances many others of our own--if this pretended generative power were a reality, surely it must have manifested itself somewhere. Speaking roundly, I should say that in such closed chambers at least five hundred chances have been given to it, but it has nowhere appeared.

The argument is now to be clenched by an experiment which will remove every residue of doubt as to the ability of the infusions here employed to sustain life. We open the back doors of our sealed chambers, and permit the common air with its floating particles to have access to our tubes. For three months they have remained pellucid and sweet--flesh, fish, and vegetable extracts purer than ever cook manufactured. Three days' exposure to the dusty air suffices to render them muddy, fetid, and swarming with infusorial life. The liquids are thus proved, one and all, ready for putrefaction when the contaminating agent is applied. I invite my colleague to reflect on these facts. How will he account for the absolute immunity of a liquid exposed for months in a warm room to optically pure air, and its infallible putrefaction in a few days when exposed to dust-laden air? He must, I submit, bow to the conclusion that the dust-particles are the cause of putrefactive life. And unless he accepts the hypothesis that these particles, being dead in the air, are in the liquid miraculously kindled into living things, he must conclude that the life we have observed springs from germs or organisms diffused through the atmosphere.

The experiments with hermetically sealed flasks have reached the number of 940. A sample group of 130 of them were laid before the Royal Society on January 13, 1876. They were utterly free from life, having been completely sterilised by three minutes' boiling. Special care had been taken that the temperatures to which the flasks were exposed should include those previously alleged to be efficient. The conditions laid down by the heterogenist were accurately copied, but there was no corroboration of his results. Stress was then laid on the question of warmth, thirty degrees being suddenly added to the temperatures with which both of us had previously worked. Waiving all protest against the caprice thus manifested, I met this new requirement also. The sealed tubes, which had proved barren in the Royal Institution, were suspended in perforated boxes, and placed under the supervision of an intelligent assistant in the Turkish Bath in Jermyn Street. From two to six days had been allowed for the generation of organisms in hermetically sealed tubes. Mine remained in the washing-room of the bath for nine days. Thermometers placed in the boxes, and read off twice or three times a day, showed the temperature to vary from a minimum of 101° to a maximum of 112° Fahr. At the end of nine days the infusions were as clear as at the beginning. They were then removed to a warmer position. A temperature of 115° had been mentioned as particularly favourable to spontaneous generation. For fourteen days the temperature of the Turkish Bath hovered about this point, falling once as low as 106°, reaching 116° on three occasions, 118° on one, and 119° on two. The result was quite the same as that just recorded. The higher temperatures proved perfectly incompetent to develope life.

Taking the actual experiment we have made as a basis of calculation, if our 940 flasks were opened on the hayloft of the Bel Alp, 858 of them would become filled with organisms. The escape of the remaining 82 strengthens our case, proving as it does conclusively that not in the air, nor in the infusions, nor in anything continuous diffused through the air, but in discrete particles, suspended in the air and nourished by the infusions, we are to seek the cause of life. Our experiment proves these particles to be in some cases so far apart on the hayloft as to permit 10 per cent of our flasks to take in air without contracting contamination. A quarter of a century ago Pasteur proved the cause of 'so-called spontaneous generation' to be discontinuous. I have already referred to his observation that 12 out of 20 flasks opened on the plains escaped infection, while 19 out of 20 flasks opened on the Mer de Glace escaped. Our own experiment at the Bel Alp is a more emphatic instance of the same kind, 90 per cent of the flasks opened in the hayloft being smitten, while not one of those opened on the free mountain ledge was attacked.

The power of the air as regards putrefactive infection is incessantly changing through natural causes, and we are able to alter it at will. Of a number of flasks opened in 1876 in the laboratory of the Royal Institution, 42 per cent. were smitten, while 58 per cent. escaped. In 1877 the proportion in the same laboratory was 68 per cent. smitten, to 32 intact. The greater mortality, so to speak, of the infusions in 1877 was due to the presence of hay which diffused its germinal dust in the laboratory air, causing it to approximate as regards infective virulence to the air of the Alpine loft. I would ask my friend to bring his scientific penetration to bear upon all the foregoing facts. They do not prove spontaneous generation to be 'impossible.' My assertions, however, relate not to 'possibilities,' but to proofs, and the experiments just described do most 'distinctly prove the evidence on which the heterogenist relies to be written on waste paper.

My colleague will not, I am persuaded, dispute these results; but he may be disposed to urge that other able and honourable men working at the same subject have arrived at conclusions different from mine. Most freely granted; but let me here recur to the remarks already made in speaking of the experiments of Spallanzani, to the effect that the failure of others to confirm his results by no means upsets their evidence. To fix the ideas, let us suppose that my colleague comes to the laboratory of the Royal Institution, repeats there my experiments, and obtains confirmatory results; and that he then goes to University or King's College where, operating with the same infusions, he obtains contradictory results. Will he be disposed to conclude that the selfsame substance is barren in Albemarle Street and fruitful in Gower Street or the Strand? His Alpine experience has already made known to him the literally infinite differences existing between different samples of air as regards their capacity for putrefactive infection. And, possessing this knowledge, will he not substitute for the adventurous conclusion that an organic infusion is barren at one place and spontaneously generative at another, the more rational and obvious one that the atmospheres of the two localities which have had access to the infusion are infective in different degrees?

As regards workmanship, moreover, he will not fail to bear in mind, that fruitfulness may be due to errors of manipulation, while barrenness involves the presumption of correct experiment. It is only the careful worker that can secure the latter, while it is open to every novice to obtain the former. Barrenness is the result at which the conscientious experimenter, whatever his theoretic convictions may be, ought to aim, omitting no pains to secure it, and resorting only when there is no escape from it to the conclusion that the life observed comes from no source which correct experiment could neutralise or avoid.

Let us again take a definite case. Supposing my colleague to operate with the same apparent care on 100 infusions--or rather on 100 samples of the same infusion--and that 50 of them prove fruitful and 50 barren. Are we to say that the evidence for and against heterogeny is equally balanced? There are some who would not only say this, but who would treasure up the 50 fruitful flasks as 'positive' results, and lower the evidential value of the 50 barren flasks by labelling them 'negative' results. This, as shown by Dr. William Roberts, is an exact inversion of the true order of the terms positive and negative. [Footnote: See his truly philosophical remarks on this head in the 'British Medical Journal,' 1876, p. 282.] Not such, I trust, would be the course pursued by my friend. As regards the 50 fruitful flasks he would, I doubt not, repeat the experiment with redoubled care and scrutiny, and not by one repetition only, but by many, assure himself that he had not fallen into error. Such faithful scrutiny fully carried out would infallibly lead him to the conclusion that here, as in all other cases, the evidence in favour of spontaneous generation crumbles in the grasp of the competent enquirer.

The botanist knows that different seeds possess different powers of resistance to heat. [Footnote: I am indebted to Dr. Thiselton Dyer for various illustrations of such differences. It is, however, surprising that a subject of such high scientific importance should not have been more thoroughly explored. Here the scoundrels who deal in killed seeds might be able to add to our knowledge.] Some are killed by a momentary exposure to the boiling temperature, while others withstand it for several hours. Most of our ordinary seeds are rapidly killed, while Pouchet made known to the Paris Academy of Sciences in 1866, that certain seeds, which had been transported in fleeces of wool from Brazil, germinated after four hours' boiling. The germs of the air vary as much among themselves as the seeds of the botanist. In some localities the diffused germs are so tender that boiling for five minutes, or even less, would be sure to destroy them all; in other localities the diffused germs are so obstinate, that many hours' boiling would be requisite to deprive them of their power of germination. The absence or presence of a truss of desiccated hay would produce differences as great as those here described. The greatest endurance that I have ever observed--and I believe it is the greatest on record--was a case of survival after eight hours' boiling.

As regards their power of resisting heat, the infusorial germs of our atmosphere might be classified under the following and intermediate heads: Killed in five minutes; not killed in five minutes but killed in fifteen; not killed in fifteen minutes but killed in thirty; not killed in thirty minutes but killed in an hour; not killed in an hour but killed in two hours; not killed in two but killed in three hours; not killed in three but killed in four hours. I have had several cases of survival after four and five hours' boiling, some survivals after six, and one after eight hours' boiling. Thus far has experiment actually reached; but there is no valid warrant for fixing upon even eight hours as the extreme limit of vital resistance. Probably more extended researches (though mine have been very extensive) would reveal germs more obstinate still. It is also certain that we might begin earlier, and find germs which are destroyed by a temperature far below that of boiling water. In the presence of such facts, to speak of a death-point of bacteria and their germs would be unmeaning--but of this more anon.

'What present warrant,' it has been asked, 'is there for supposing that a naked, or almost naked, speck of protoplasm can withstand four, six, or eight hours' boiling?' Regarding naked specks of protoplasm I make no assertion. I know nothing about them, save as the creatures of fancy. But I do affirm, not as a 'supposition,' nor an 'assumption,' nor a 'probable guess,' nor as 'a wild hypothesis,' but as a matter of the most undoubted fact, that the spores of the hay bacillus, when thoroughly desiccated by age, have withstood the ordeal mentioned. And I further affirm that these obdurate germs, under the guidance of the knowledge that they are germs, can be destroyed by five minutes' boiling, or even less. This needs explanation. The finished bacterium perishes at a temperature far below that of boiling water, and it is fair to assume that the nearer the germ is to its final sensitive condition the more readily will it succumb to heat. Seeds soften before and during germination. This premised, the simple description of the following process will suffice to make its meaning understood.

An infusion infected with the most powerfully resistent germs, but otherwise protected against the floating matters of the air, is gradually raised to its boiling-point. Such germs as have reached the soft and plastic state immediately preceding their development into bacteria are thus destroyed. The infusion is then put aside in a warm room for ten or twelve hours. If for twenty-four, we might have the liquid charged with well-developed bacteria. To anticipate this, at the end of ten or twelve hours we raise the infusion a second time to the boiling temperature, which, as before, destroys all germs then approaching their point of final development. The infusion is again put aside for ten or twelve hours, and the process of heating is repeated. We thus kill the germs in order of their resistance, and finally kill the last of them. No infusion can withstand this process if it be repeated a sufficient number of times. Artichoke, cucumber, and turnip infusions, which had proved specially obstinate when infected with the germs of desiccated hay, were completely broken down by this method of discontinuous heating, three minutes being found sufficient to accomplish what three hundred minutes' continuous boiling failed to accomplish. I applied the method, moreover, to infusions of various kinds of hay, including those most tenacious of life. Not one of them bore the ordeal. These results were clearly foreseen before they were realised, so that the germ theory fulfils the test of every true theory, that test being the power of prevision.

When 'naked or almost naked specks of protoplasm' are spoken of, the imagination is drawn upon, not the objective truth of Nature. Such words sound like the words of knowledge where knowledge is really nil. The possibility of a 'thin covering' is conceded by those who speak in this way. Such a covering may, however, exercise a powerful protective influence. A thin pellicle of India-rubber, for example, surrounding a pea keeps it hard in boiling water for a time sufficient to reduce an uncovered pea to a pulp. The pellicle prevents imbibition, diffusion, and the consequent disintegration. A greasy or oily surface, or even the layer of air which clings to certain bodies, would act to some extent in a similar way. 'The singular resistance of green vegetables to sterilisation,' says Dr. William Roberts, 'appears to be due to some peculiarity of the surface, perhaps their smooth glistening epidermis which prevented complete wetting of their surfaces.' I pointed out in 1876 that the process by which an atmospheric germ is wetted would be an interesting subject of investigation. A dry microscope covering-glass may be caused to float on water for a year. A sewing-needle may be similarly kept floating, though its specific gravity is nearly eight times that of water.

Were it not for some specific relation between the matter of the germ and that of the liquid into which it falls, wetting would be simply impossible. Antecedent, to all development there must be an interchange of matter between the germ and its environment; and this interchange must obviously depend upon the relation of the germ to its encompassing liquid. Anything that hinders this interchange retards the destruction of the germ in boiling water. In my paper published in the 'Philosophical Transactions' for 1877, I add the following remark:

It is not difficult to see that the surface of a seed or germ may be so affected by desiccation and other causes as practically to prevent contact between it and the surrounding liquid. The body of a germ, moreover, may be so indurated by time and dryness as to resist powerfully the insinuation of water between its constituent molecules. It would be difficult to cause such a germ to imbibe the moisture necessary to produce the swelling and softening which precede its destruction in a liquid of high temperature.

*****

However this may be--whatever be the state of the surface, or of the body, of the spores of Bacillus subtilis, they do as a matter of certainty resist, under some circumstances, exposure for hours to the heat of boiling water. No theoretic scepticism can successfully stand in the way of this fact, established as it has been by hundreds, if not thousands, of rigidly conducted experiments.

*****

We have now to test one of the principal foundations of the doctrine of spontaneous generation as formulated in this country. With this view, I place before my friend and co-enquirer two liquids which have been kept for six months in one of our sealed chambers, exposed to optically pure air. The one is a mineral solution containing in proper proportions all the substances which enter into the composition of bacteria, the other is an infusion of turnip-it might be any one of a hundred other infusions, animal or vegetable. Both liquids are as clear as distilled water, and there is no trace of life in either of them. They are, in fact, completely sterilised. A mutton-chop, over which a little water has been poured to keep its juices from drying up, has lain for three days upon a plate in our warm room. It smells offensively. Placing a drop of the fetid mutton-juice under a microscope, it is found swarming with the bacteria of putrefaction. With a speck of the swarming liquid I inoculate the clear mineral solution and the clear turnip infusion, as a surgeon might inoculate an infant with vaccine lymph. In four-and-twenty hours the transparent liquids have become turbid throughout, and instead of being barren as at first they are teeming with life. The experiment may be repeated a thousand times with the same invariable result. To the naked eye the liquids at the beginning were alike, being both equally transparent-to the naked eye they are alike at the end, being both equally muddy. Instead of putrid mutton-juice, we might take as a source of infection any one of a hundred other putrid liquids, animal or vegetable. So long as the liquid contains living bacteria a speck of it communicated either to the clear mineral solution, or to the clear turnip infusion, produces in twenty-four hours the effect here described.

We now vary the experiment thus: Opening the back-door of another closed chamber which has contained for months the pure mineral solution and the pure turnip infusion side by side, I drop into each of them a small pinch of laboratory dust. The effect here is tardier than when the speck of putrid liquid was employed. In three days, however, after its infection with the dust, the turnip infusion is muddy, and swarming as before with bacteria. But what about the mineral solution which, in our first experiment, behaved in a manner undistinguishable from the turnip-juice? At the end of three days there is not a bacterium to be found in it. At the end of three weeks it is equally innocent of bacterial life. We may repeat the experiment with the solution and the infusion a hundred times with the same invariable result. Always in the case of the latter the sowing, of the atmospheric dust yields a crop of bacteria-never in the former does the dry germinal matter kindle into active life. [Footnote: This is the deportment of the mineral solution as described by others. My own experiments would lead me to say that the development of the bacteria, though exceedingly slow and difficult, is not impossible.] What is the inference which the reflecting mind must draw from this experiment? Is it not as clear as day that while both liquids are able to feed the bacteria and to enable them to increase and multiply, after they have been once, fully developed, only one of the liquids is able to develope into active bacteria the germinal dust of the air?

I invite my friend to reflect upon this conclusion he will, I think, see that there is no escape from it. He may, if he prefers, hold the opinion, which I consider erroneous, that bacteria exist in the air, not as germs but as desiccated organisms. The inference remains, that while the one liquid is able to force the passage from the inactive to the active state, the other is not.

But this is not at all the inference which has been drawn from experiments with the mineral solution.

Seeing its ability to nourish bacteria when once inoculated with the living active organism, and observing that no bacteria appeared in the solution after long exposure to the air, the inference was drawn that neither bacteria nor their germs existed in the air. Throughout Germany the ablest literature of the subject, even that opposed to heterogeny, is infected with this error; while heterogenists at home and abroad have based upon it a triumphant demonstration of, their doctrine. It is proved, they say, by the deportment of the mineral solution that neither bacteria nor their germs exist in the air; hence, if, on exposing a thoroughly sterilised turnip infusion to the air, bacteria appear, they must of necessity have been spontaneously generated. In the words of Dr. Bastian: 'We can only infer that whilst the boiled saline solution is quite incapable of engendering bacteria, such organisms are able to arise _de novo_ in the boiled organic infusion.' [Footnote: 'Proceedings of the Royal Society,' vol. xxi. p. 130.]

I would ask my eminent colleague what he thinks of this reasoning now? The datum is--'A mineral solution exposed to common air does not develope bacteria;' the inference is--'Therefore if a turnip infusion similarly exposed develope bacteria, they must be spontaneously generated.' The inference, on the face of it, is an unwarranted one. But while as matter of logic it is inconclusive, as matter of fact it is chimerical. London air is as surely charged with the germs of bacteria as London chimneys are with smoke. The inference just referred to is completely disposed of by the simple question: 'Why, when your sterilised organic infusion is exposed to optically puree air, should this generation of life _de novo_ utterly cease? Why should I be able to preserve my turnip-juice side by side with your saline solution for the three hundred and sixty-five days of the year, in free connection with the general atmosphere, on the sole condition that the portion of that atmosphere in contact with the juice shall be visibly free from floating dust, while three days' exposure to that dust fills it with bacteria?' Am I over sanguine in hoping that as regards the argument here set forth he who runs may read, and he who reads may understand?