Supposing an infusion intrinsically barren, but readily susceptible of putrefaction when exposed to common air, to be brought into contact with this unilluminable air, what would be the result? It would never putrefy. It might, however, be urged that the air is spoiled by its violent calcination. Oxygen passed through a spirit-lamp flame is, it may be thought, no longer the oxygen suitable for the development and maintenance of life. We have an easy escape from this difficulty, which is based, however, upon the unproved assumption that the air has been affected by the flame. Let a condensed beam be sent through a large flask or bolthead containing common air. The track of the beam is seen within the flask — the dust revealing the light, and the light revealing the dust. Cork the flask, stuff its neck with cotton-wool, or simply turn it mouth downwards and leave it undisturbed for a day or two. Examined afterwards with the luminous beam, no track is visible; the light passes through the flask as through a vacuum. The floating matter has abolished itself, being now attached to the interior surface of the flask.
Were it our object, as it will be subsequently, to effectually detain the dirt, we might coat that surface with some sticky substance. Here, then, without 'torturing' the air in any way, we have found a means of ridding it, or rather of enabling it to rid itself, of floating matter.
We have now to devise a means of testing the action of such spontaneously purified air upon putrescible infusions. Wooden chambers, or cases, are accordingly constructed, having glass fronts, side-windows, and back-doors. Through the bottoms of the chambers test-tubes pass air-tight; their open ends, for about one-fifth of the length of the tubes, being within the chambers. Provision is made for a free connection rough sinuous channels between the inner and the outer air. Through such channels, though open, no dust will reach the chamber. The top of each chamber is perforated by a circular hole two inches in diameter, closed air-tight by a sheet of India-rubber. This is pierced in the middle by a pin, and through the pin-hole is pushed the shank of a long pipette, ending above in a small funnel. The shank also passes through a stuffing-box of cotton-wool moistened with glycerine; so that, tightly clasped by the rubber and wool, the pipette is not likely in its motions up and down to carry any dust into the chamber. The annexed woodcut shows a chamber, with six test-tubes, its side-windows w w, its pipette p c, and its sinuous channels a b which connect the air of the chamber with the outer air.
The chamber is carefully closed and permitted to remain quiet for two or three days. Examined at the beginning by a beam sent through its windows, the air is found laden with floating matter, which in three days has wholly disappeared. To prevent its ever rising again, the internal surface of the chamber was at the outset coated with glycerine. The fresh but putrescible liquid is introduced into the six tubes in succession by means of the pipette. Permitted to remain without further precaution, every one of the tubes would putrefy and fill itself with life. The liquid has been in contact with the dust-laden air outside by which it has been infected, and the infection must be destroyed. This is done by plunging the six tubes into a bath of heated oil and boiling the infusion. The time requisite to destroy the infection depends wholly upon its nature. Two minutes' boiling suffices to destroy some contagia, whereas two hundred minutes' boiling fails to destroy others. After the infusion has been sterilised, the oil-bath is withdrawn, and the liquid, whose putrescibility has been in no way affected by the boiling, is abandoned to the air of the chamber.
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.