The life of Pasteur

by René Vallery-Radot

The confusion of ideas on the origin of contagious and epidemic diseases was about to be suddenly enlightened; Pasteur had now taken up the study of the disease known as charbon or splenic fever. This disease was ruining agriculture; the French provinces of Beauce, Brie, Burgundy, Nivernais, Berry, Champagne, Dauphiné and Auvergne, paid a formidable yearly tribute to this mysterious scourge. In the Beauce, for instance, twenty sheep out of every hundred died in one flock; in some parts of Auvergne the proportion was ten or fifteen per cent., sometimes even twenty-five, thirty-five, or fifty per cent. At Provins, at Meaux, at Fontainebleau, some farms were called charbon farms; elsewhere, certain fields or hills were looked upon as accursed and an evil spell seemed to be thrown over flocks bold enough to enter those fields or ascend those hills. Animals stricken with this disease almost always died in a few hours; sheep were seen to lag behind the flock, with drooping head, shaking limbs and gasping breath; after a rigor and some sanguinolent evacuations, occurring also through the mouth and nostrils, death supervened, often before the shepherd had had time to notice the attack. The carcase rapidly became distended, and the least rent in the skin gave issue to a flow of black, thick and viscid blood, hence the name of anthrax given to the disease. It was also called splenic fever, because necropsy showed that the spleen had assumed enormous dimensions; if that were opened, it presented a black and liquid pulp. In some places the disease assumed a character of extreme virulence; in the one district of Novgorod, in Russia, 56,000 head of cattle died of splenic infection between 1867 and 1870. Horses, oxen, cows, sheep, everything succumbed, as did also 528 persons, attacked by the contagion under divers forms; a pin prick or a scratch is sufficient to inoculate shepherds, butchers, knackers or farmers with the malignant pustule.

Though a professor at the Alfort Veterinary School, M. Delafond, did point out to his pupils as far back as 1838 that charbon blood contained “little rods,” as he called them; it was only looked upon by himself and them as a curiosity with no scientific importance. Davaine, when he—and Rayer as well—recognized in 1850 those little filiform bodies in the blood of animals dying of splenic fever, he too merely mentioned the fact, which seemed to him of so little moment that he did not even report it in the first notice of his works edited by himself.

It was only eleven years later that Davaine—struck, as he himself gladly acknowledged, by reading Pasteur’s paper on the butyric ferment, the little cylindrical rods of which offer all the characteristics of vibriones or bacteria—asked himself whether the filiform corpuscles seen in the blood of the charbon victims might not act after the manner of ferments and be the cause of the disease. In 1863, a medical man at Dourdan, whose neighbour, a farmer, had lost twelve sheep of charbon in a week, sent blood from one of these sheep to Davaine, who hastened to inoculate some rabbits with this blood. He recognized the presence of those little transparent and motionless rods which he called bacteridia (a diminutive of bacterium, or rod-shaped vibriones). It might be thought that the cause of the evil was found, in other words that the relation between those bacteridia and the disease which had caused death could not be doubted. But two professors of the Val de Grâce, Jaillard and Leplat; refuted these experiments.

They had procured, in the middle of the summer, from a knacker’s yard near Chartres, a little blood from a cow which had died of anthrax, and they inoculated some rabbits with it. The rabbits died, but without presenting any bacteridia. Jaillard and Leplat therefore affirmed that splenic fever was not an affection caused by parasites, that the bacteridium was an epiphenomenon of the disease and could not be looked upon as the cause of it.

Davaine, on repeating Jaillard and Leplat’s experiments, found a new interpretation; he alleged that the disease they had inoculated was not anthrax. Then Jaillard and Leplat obtained a little diseased sheep’s blood from M. Boutet, a veterinary surgeon at Chartres, and tried that instead of cow’s blood. The result was identical: death ensued, but no bacteridia. Were there then two diseases?

Others made observations in their turn. It occurred to a young German physician, Dr. Koch, who in 1876 was beginning his career in a small village in Germany, to seek a culture medium for the bacteridium. A few drops of aqueous humour, collected in the eyes of oxen or of rabbits, seemed to him favourable. After a few hours of this nutrition the rods seen under the microscope were ten or twenty times larger than at first; they lengthened immoderately, so as to cover the whole slide of the microscope, and might have been compared to a ball of tangled thread. Dr. Koch examined those lengths, and after a certain time noticed little spots here and there looking like a punctuation of spores. Tyndall, who knew how to secure continuous attention by a variety of comparisons, said at a scientific conference in Glasgow a few months later that those little ovoid bodies were contained within the envelope of the filament like peas in their pods. It is interesting to note that Pasteur, when he studied, in connection with silkworm diseases, the mode of reproduction of the vibriones of flachery, had seen them divide into spores similar to shining corpuscles; he had demonstrated that those spores, like seeds of plants, could revive after a lapse of years and continue their disastrous work. The bacterium of charbon, or bacillus anthracis as it now began to be called, reproduced itself in the same way, and, when inoculated by Dr. Koch into guinea-pigs, rabbits and mice, provoked splenic fever as easily and inevitably as blood from the veins of an animal that had died of the disease. Bacilli and spores therefore yielded the secret of the contagion, and it seemed that the fact was established, when Paul Bert, in January, 1877, announced to the Société de Biologie that it was “possible to destroy the bacillus anthracis in a drop of blood by compressed oxygen, to inoculate what remained, and to reproduce the disease and death without any trace of the bacteridium ... Bacteridia,” he added, “are therefore neither the cause nor the necessary effect of splenic fever, which must be due to a virus.”

Pasteur tackled the subject. A little drop of the blood of an animal which had died of anthrax—a microscopic drop—was laid, sown, after the usual precautions to ensure purity, in a sterilized balloon which contained neutral or slightly alkaline urine. The culture medium might equally be common household broth, or beer-yeast water, either of them neutralized by potash. After a few hours, a sort of flake was floating in the liquid; the bacteridia could be seen, not under the shape of short broken rods, but with the appearance of filaments, tangled like a skein; the culture medium being highly favourable, they were rapidly growing longer. A drop of that liquid, abstracted from the first vessel, was sown into a second vessel, of which one drop was again placed into a third, and so on, until the fortieth flask; the seed of each successive culture came from a tiny drop of the preceding one. If a drop from one of those flasks was introduced under the skin of a rabbit or guinea-pig, splenic fever and death immediately ensued, with the same symptoms and characteristics as if the original drop of blood had been inoculated. In the presence of the results from those successive cultures, what became of the hypothesis of an inanimate substance contained in the first drop of blood? It was now diluted in a proportion impossible to imagine. It would therefore be absurd, thought Pasteur, to imagine that the last virulence owed its power to a virulent agent existing in the original drop of blood; it was to the bacteridium, multiplied in each culture, and to the bacteridium alone, that this power was due; the life of the bacteridium had made the virulence. “Anthrax is therefore” Pasteur declared, “the disease of the bacteridium, as trichinosis is the disease of the trichina, as itch is the disease of its special acarus, with this circumstance, however, that, in anthrax, the parasite can only be seen through a microscope, and very much enlarged.” After the bacteridium had presented those long filaments, within a few hours, two days at the most, another spectacle followed; amidst those filaments, appeared the oval shapes, the germs, spores or seeds, pointed out by Dr. Koch. Those spores, sown in broth, reproduced in their turn the little packets of tangled filaments, the bacteridia. Pasteur reported that “one single germ of bacteridium in the drop which is sown multiplies during the following hours and ends by filling the whole liquid with such a thickness of bacteridia that, to the naked eye, it seems that carded cotton has been mixed with the broth.”

M. Chamberland, a pupil who became intimately associated with this work on anthrax, has defined as follows what Pasteur had now achieved: “By his admirable process of culture outside organism, Pasteur shows that the rods which exist in the blood, and for which he has preserved the name of bacteridia given them by Davaine, are living beings capable of being indefinitely reproduced in appropriate liquids, after the manner of a plant multiplied by successive cuttings. The bacterium does not reproduce itself only under the filamentous form, but also through spores or germs, after the manner of many plants which present two modes of reproduction, by cuttings and by seeds.” The first point was therefore settled. The ground suspected and indicated by Davaine was now part of the domain of science, and preserved from any new attacks.

Yet Jaillard and Leplat’s experiments remained to be explained: how had they provoked death through the blood of a splenic fever victim and found no bacteridia afterwards? It was then that Pasteur, guided, as Tyndall expressed it, by “his extraordinary faculty of combining facts with the reasons of those facts,” placed himself, to begin with, in the conditions of Jaillard and Leplat, who had received, during the height of the summer, some blood from a cow and a sheep which had died of anthrax, that blood having evidently been abstracted more than twenty-four hours before the experiment. Pasteur, who had arranged to go to the very spot, the knacker’s yard near Chartres, and himself collect diseased blood, wrote to ask that the carcases of animals which had died of splenic fever should be kept for him for two or three days.

He arrived on June 13, 1877, accompanied by the veterinary surgeon, M. Boutet. Three carcases were awaiting him: that of a sheep which had been dead sixteen hours, that of a horse whose death dated from the preceding day, and that of a cow which must have been dead for two or three days, for it had been brought from a distant village. The blood of the recently diseased sheep contained bacteridia of anthrax only. In the blood of the horse, putrefaction vibriones were to be found, besides the bacteridia, and those vibriones existed in a still greater proportion in the blood of the cow. The sheep’s blood, inoculated into guinea-pigs, provoked anthrax with pure bacteridia; that of the cow and of the horse brought a rapid death with no bacteridia.