Louis Pasteur 6

	Louis Pasteur 6

Following up the idea that a connection between the disease and the corpuscles might possibly exist, as other observers had previously imagined, Pasteur declared, in a Note presented to the Agricultural Committee of Alais on June 26, 1865, twenty days after his arrival, that it was a mistake to seek for the corpuscle in the eggs or in the worms. Both the one and the other could carry in them the germ of the disease, without exhibiting distinct corpuscles, visible under the microscope. The evil developed itself especially in the chrysalides and in the moths, and it was in them that search should be made. Finally, Pasteur came to the conclusion that the only infallible method of procuring healthy eggs must be by having recourse to moths free from corpuscles. Pasteur hastened to apply this new method of obtaining pure eggs. Notwithstanding that the malady was universally prevalent, he succeeded, after several days of assiduous microscopic observations, in finding some moths free from corpuscles. He carefully preserved their eggs, as well as other eggs which had proceeded from very corpusculous couples, intending to wait for what these eggs would produce the following year; the first would be probably free from corpuscles, while the latter would contain them. He would thus have in future, though on a small scale, samples of originally healthy and of originally unhealthy cultivations, by the comparison of which with the cultivations of the trade--all more or less smitten with the evil--totally new views might be expected to emerge. Who can tell, thought Pasteur, whether the prosperity of the silk cultivation may not depend on the practical application of this production of pure eggs by means of moths free from corpuscles?                    *       *       *       *       * Scarcely had Pasteur made known, first to the Committee of Alais, and then to the Academy of Sciences, the results of his earliest observations and the inductions to which they pointed, when critics without number arose on all sides. It was objected that the labours of several Italian _savants_ had established beyond all doubt that the corpuscles were a normal element of certain worms, and especially of all the moths when old; that other authors had affirmed it to be sufficient to starve certain worms to make these famous corpuscles appear in all their tissues; and that Dr. Gaetano Cantoni had already tried some cultivations with eggs coming from moths without corpuscles, and that he had totally failed. 'Your efforts will be vain,' wrote the celebrated Italian entomologist Cornalia; 'your selected eggs will produce healthy worms, but these worms will become sickly through the influence of the epidemic demon which reigns everywhere.' Anyone but Pasteur would have been staggered, but he was not the man to allow himself to be discouraged by _a priori_ opinions, and by assertions which were more or less guesswork. He was resolved not to abandon his preconceived idea until experiment had pronounced upon it with precision. All scientific research, in order to be undertaken and followed up with success, should have, as point of departure, a preconceived idea, an hypothesis which we must seek to verify by experiment. To judge of the value of the facts which Pasteur had just announced, it was necessary to know if there existed the relation of cause and effect between the corpuscles and the disease. This was the great point to be elucidated.                    *       *       *       *       * But if, without preliminary groping, he had discovered the way to be pursued, Pasteur subsequently brought to bear his rare prudence as an experimentalist. For five years he returned annually for some months to Alais. The little house nestling among the trees called Pont-Guisquet became at the same time his habitation and his silkworm nursery. It is hemmed in by mountains, up the sides of which terraces rise, one above the other, planted with mulberry trees. The solitude was profound. Madame Pasteur and her daughter constituted themselves silkworm-rearers--performing their part in earnest, not only gathering the leaves of the mulberry trees, but also taking part in all the experiments. The assistants of the Ecole Normale, Duclaux, Maillot, Gernez, and Raulin, grouped themselves around their master. Thus, in an out-of-the-way corner of the Cevennes was formed a colony seeking with ardour the solution of an obscure problem, and the means of curing or preventing a disease which had for so long a time blighted one of the great sources of the national wealth. One of the first cares of Pasteur was to settle the question as to the contagion of the disease. Many hypotheses had been formed regarding this contagion, but few experiments had been made, and none of them were decisive. Opinions were also very much divided. Some considered that contagion was certain; the majority, however, either doubted or denied its existence; some considered it as accidental. It was said, for example, that the evil was not contagious by itself, but that it became so through the presence and complication of other diseases which were themselves contagious. This hypothesis was convenient, and it enabled contradictory facts to be explained. If some persons had seen healthy worms, which had been mixed up either by mistake or intention with sickly ones, perish, and if they insisted on contagion, others forthwith replied by diametrically opposite observations. But whatever the divergences of opinion might be, everyone at all events believed in the existence of a poisonous medium rendered epidemic by some occult influence. Pasteur soon succeeded, by accurate experiments, in proving absolutely that the evil was contagious. One of the first experiments was as follows. After their first moulting, he took some very sound worms free from corpuscles, and fed them with corpusculous matter, which he prepared in the following simple manner. He pounded up a silkworm in a little water, and passed a paint-brush dipped in this liquid over the whole surface of the leaves. During several days there was not the least appearance of disease in the worms fed on those leaves; they reached their second moulting at the same time as the standard worms which had not been infected. The second moulting was accomplished without any drawback. This was a proof that all the worms, those infected as well as the standard lot, had taken the same amount of nourishment. The parasite was apparently not present. Matters remained in this state for some days longer. Even the third moulting was got through without any marked difference between the two groups of worms. But soon important changes set in. The corpuscles, which had hitherto only showed themselves in the integuments of the intestines, began to appear in the other organs. From the second day following the third moulting--that is to say, the twelfth after the infection--a visible inequality distinguished the infected from the non-infected worms. Those of the standard lot were clearly in much the best health. On examining the infected worms through a magnifying glass, a multitude of little spots were discovered on their heads, and on the rings of their bodies, which had not before shown themselves. These spots appeared on the exterior skin when the interior skin of the intestinal canal contained a considerable number of corpuscles. It was these corpuscles that impeded the digestive functions, and interfered with the assimilation of the food. Hence arose the inequality of size of the worms. After the fourth moulting, the same type of disease was noticed as that which was breaking out everywhere in the silkworm nurseries, especially the symptom of spots on the skin, which had led to the disease being called _pebrine_. The peasants said that the worms were peppered. The majority of the worms were full of corpuscles. Those which spun their cocoons produced chrysalides which were nothing but corpusculous pulp, if such a term be allowed. It was thus proved that the corpuscles, introduced into the intestinal canal at the same time as the food of the worms, convey the infection into the intestinal canal, and progressively into all the tissues. The malady had in certain cases a long period of incubation, since it was only on the twelfth day that it became perceptible. Finally, the spots of _pebrine_ on the skin, far from being the disease itself, were but the effect of the corpuscles developed in the interior; they were but a sign, already removed from the true seat of the evil. 'If these spots of _pebrine_,' thought Pasteur, 'were considered in conjunction with certain human maladies in which spots and irruptions appear on the body, what interesting inductions might present themselves to minds prepared to receive them!' Pasteur was never tired of repeating this curious experiment, or of varying its conditions. Sometimes he introduced the corpusculous food into healthy worms at their birth, sometimes at the second or third moulting. Occasionally, when the worms were about to spin their cocoons, the corpusculous food was given them. All the disasters that were known to have happened in the silkworm nurseries, their extent and their varied forms, were faithfully reproduced. Pasteur created at will any required manifestation of _pebrine_. When he infected quite healthy worms, after their fourth moulting, with fresh corpusculous matter, these worms, even after several meals of corpusculous leaves alternated with meals of wholesome leaves, made their cocoons. It might have been supposed that in this case the contagion had not taken effect. This was but a deceptive appearance. The communication of the disease exhibited itself in a marked degree in the chrysalides and in the moths. Many of the chrysalides died before they turned into moths, and their bodies might be said to be entirely composed of corpuscles. Such moths as were formed, and which emerged from their cocoons, had a most miserable appearance. The disease sometimes went so far as to render breeding and the laying of eggs impossible. Faithful to the rules prescribed by the experimental method, Pasteur was careful to reproduce these same experiments with the worms of the standard lot, from which all infected worms had been selected. He fed these healthy worms on leaves over which a clear infusion made from the remains of moths or worms exempt from corpuscles had been spread with a paint-brush, instead of leaves contaminated with corpusculous remains. This food kept the worms in their usual health. Could there be a better proof that the corpuscles alone were the real cause of the _pebrine_ disease? These experiments, I repeat, threw a strong light on the nature of the disease, and exactly accounted for what took place in the industrial cultivations. From the malady which attacked the worms at their birth, decimating a whole cultivation, down to the invisible disease that may be said to inclose itself in the cocoon, all was now explained. One of the effects of the plague which had most excited the surprise and thwarted the efforts of cultivators was the impossibility of finding productive eggs, even when they tried to obtain them from the cocoons of groups which had succeeded perfectly well as far as the production of cocoons was concerned. It was proved that almost invariably the following year the eggs of these fine-looking groups were unproductive. Numbers of the agricultural boards, and practitioners, not being able to believe in the existence of the disease in collections that were so satisfactory as regards the abundance and beauty of the cocoons, persisted in thinking that the failures had an origin not connected with the seed itself. This resulted in deception after deception, often even in mistakes that were much to be regretted. Frequently the best husbandmen were known to reserve for the production of eggs some very fine cultivations, not having observed in the worms either spots of _pebrine_ or corpuscles even up to the time when the mounting of the brambles had been accomplished; and the year following they had the pain of seeing all the cultivations from these eggs perish. These circumstances, so well calculated to produce discouragement and to give the disease a mysterious character, met with their natural explanation in the facts proved by experimental infection. Still, as it never occurs to the cultivator to infect the worms directly by giving them food charged with corpusculous debris, it might be asked how, in the industrial establishments, such results can be produced. Pasteur lost no time in solving this difficulty. In a cultivation containing corpusculous worms these worms perpetually furnish contagious matter, which falls upon the leaf and fouls it. This is the excreta of the worms, which the microscope shows to be more or less filled with corpuscles drawn from the lining of the intestinal canal. It is there that they swarm. It is easy to understand that these excreta, falling on the leaves, contaminate them all the more easily because the worms, by the weight of their bodies in crawling, press the excreta against the leaves. This is one cause of natural contagion. By the excreta of corpusculous worms which he crushed, mixed with water, and spread with a paint-brush over the mulberry leaves intended for a single meal, Pasteur was able to communicate the contagion to as many worms as he liked. He also indicated another natural and direct cause of contagion. The six fore-feet of the worm have sharp hooks at their ends, by means of which the worms prick each other's skins. Let any one imagine a healthy worm passing over the body of the corpusculous worm. The hooks of the first worm, by penetrating the skin of the second, are liable to be soiled by the corpuscles immediately below that skin; and these hooks are capable of carrying the seeds of disease to other healthy worms, which may be pricked in their turn. To demonstrate experimentally, as Pasteur did, the existence of this cause of contagion, it was only necessary to take some worms and allow them to wound each other. Lastly, infection at a distance, through the medium of the air and the dust it carries, is a fact equally well established. It is sufficient, by sweeping the breeding-houses, or by shaking the hurdles, to stir up the dust of corpusculous excretions and the dried remains of dead worms, and to allow them to be spread over the hurdles of the healthy worms, to cause, after a certain time, contagion to appear among these worms. When very healthy worms were placed in a breeding nursery at a considerable distance from unhealthy worms, they, in their turn, became infected. After so many decisive experiments it was no longer possible not to see in _pebrine_ an essentially contagious disease. Nevertheless, among facts invoked in favour of non-contagion, there was one which it was difficult to explain. There existed several examples of successful cultivations conducted in nurseries which had totally failed from the effects of _pebrine_ the year before. The explanation is, as shown by Pasteur, that the dust can only act as a contagion when it is fresh. Corpusculous matter, when thoroughly dried, loses its virulence. A few weeks suffice to render such matter inoffensive: hence the dust of one year is not injurious to the cultivations of the next year. The corpuscles contained in the eggs intended for future cultivation alone cause the transmission of the disease to future generations. And what can be more easily understood than the presence of corpusculous parasites in the egg? The egg comes into existence during that marvellous phase of the life of a silkworm when, after having spun its cocoon, it sleeps within it as a chrysalis, resolving itself, so to speak, into those kinds of albumen and yelk from which the fully-formed moth will emerge, as a chick emerges from its egg. Let anyone imagine this origin of an approaching life, no longer in its normal purity, but associated with a parasite which will find in the materials surrounding it, so adapted to life and transformation, the elements of its own nourishment and multiplication. This parasite will be present when the eggs of the female moth, tender and soft as albumen, begin to define their outlines. Woe betide those eggs if they then enclose any particles of corpuscle, or of its original matrix. In vain will the envelope of those eggs become by degrees hard and horny; the enemy is within, and later on he will be discovered in the embryo of the silkworm. Thus this terrible plague is at the same time contagious and hereditary, helping us to understand the evolution of this double character in certain maladies both of men and animals.                                   II. The first time Pasteur went to Alais the silkworm epidemic was universally attributed to a single cause--_pebrine_. _Pebrine_ was called _the disease_. This word expressed everything. It indicated the existence of a mysterious scourge, the origin and nature of which could not be traced, but which was ready to fall upon all the establishments devoted to the nurture of the worms. Whatever might happen, or whatever might be the cause of ruin in a silkworm nursery, _the disease_ was held responsible. One of the most striking proofs that the evil was attributed to _pebrine_ alone is found in the fact that a prize of 5,000 florins was offered by the Austrian Government in 1868, as a reward for the discovery of the best remedy for the prevention and cure of _pebrine_--'the epidemic disease which devastates the silkworm.' A rapid glance at the principles which have just been established suffices to show that _pebrine_ might now be regarded as vanquished. Pasteur had demonstrated that moths free from corpuscles never produced a single corpusculous egg; he had proved, moreover, that eggs brought up in a state of isolation, at a distance from contaminated eggs, produce no worms, chrysalides, or moths which are corpusculous. It was easy, therefore, to multiply cultivations free from _pebrine_. The production of silk and the production of eggs was thus secured. To make sure that the eggs were pure it was only necessary to have recourse to the microscopic examination of the moths which had produced them. These observations might be made by women, by young girls, even by children. It was sufficient to crush up a moth in a little water, and to put a drop of this mixture under the microscope, to see the corpuscles clearly, if they existed. It seemed, then, that the plague was got rid of. But Pasteur was not slow in recognising that the general belief in a single malady could not be justified. If the experiments of 1866 had demonstrated to him the full extent of the corpusculous malady, and had established the principles of a treatment proper for its prevention, the method he had adopted had also shown him that _pebrine_ was far from being the only cause from which the silk culture suffered. It was in 1867 that this result was obtained. From an experimental point of view, that year counted double for Pasteur. Influenced by a profound sympathy for the misery which he had witnessed during two successive years, and, at the same time, impatient to find the cause of the scourge, Pasteur, in the months of February, March, and April, in advance of the great industrial cultivations, commenced a series of experiments on worms hatched by artificial heat, and fed with mulberry leaves from a hothouse. During these forced experiments Pasteur observed that out of sixteen broods derived from non-corpusculous parents, fifteen succeeded, while the sixteenth perished almost entirely between the fourth moulting and the climbing on to the brambles. After having exhibited a most healthful appearance, the worms died suddenly. In a cultivation of 100 worms, ten, fifteen, twenty dead ones were picked up daily: these turned black, and became putrid with extraordinary rapidity, often within the space of twenty-four hours. Sometimes they were soft and flabby, like an empty, crumpled intestine. Consulting the authors who had written upon silkworms, Pasteur could not doubt that he had before his eyes a characteristic specimen of the disease called _morts-flats_, or _flacherie_. Not only were these worms free from all _pebrine_ spots, but no corpuscles were to be found in any part of their bodies. A still more significant fact was, that corpuscles were also absent from the chrysalides and the moths of those few worms which were able to spin their cocoons. Although this sample was confined to a single group of eggs derived from parents free from corpuscles, Pasteur continued to entertain doubts as to the existence of only a single disease, and also as to the necessary connection of _pebrine_ with _flacherie_. These suspicions were confirmed by his cultivations of April and May. Numerous cases of _flacherie_ presented themselves. Uncertainty was no longer possible as to the mutual independence of the two maladies--_pebrine_ and _flacherie_. The cultivations most seriously invaded by the last-mentioned disease came from eggs produced by parents free from corpuscles, and led on to reproducers also free from this parasite. On visiting a multitude of industrial cultivations, Pasteur discerned that what had passed in his own laboratory was of very general occurrence, and that, contrary to the received opinion, two distinct maladies divided between them the cause of all the misfortunes. _Pebrine_ was evidently the most widely spread, but _flacherie_ had also its share, and a very large share, in the calamity. Here, once more, the microscope came to Pasteur's assistance. If, at the period of the rearing of the silkworms, when the mean temperature is always rather high, some mulberry leaves are crushed in a mortar and mixed with a little water, the liquid being left to itself, in twenty-four hours it will be found filled with microscopic organisms; some motionless, resembling little rods or spores joined end to end, like strings of beads, others more or less active, flexible, endowed with a sinuous movement like that of the vibrios found in nearly all organic infusions in process of decomposition. Whence come these microscopic organisms? The facts relating to spontaneous generation indicate that the germs of these organisms were on the surface of the pounded leaf, spread in the form of dust over the instruments used to triturate the leaf, possibly on the mortar, the pestle, or in the water added to the pounded leaves. It is a curious fact, that if the intestinal canal of a worm in full process of digestion be opened, the pounded leaf which fills it from one end to the other will not show microscopic organisms of any kind, but only cells of parenchyma, green granules of the chlorophyl of the leaf, and remains of the air-vessels of the plant. Through the action of the liquids secreted by the glands which line the integuments of the intestinal canal, the germs of organisms are themselves digested or hindered in their development. The digestive functions of silkworms are so active that everything is carried away, destroyed in the same manner as the leaves themselves. But if from any cause the digestion of the worms be impeded or suspended, then the germs introduced with the food into the intestinal canal will give rise to the multiplication of microscopic organisms which are always found in the artificially bruised leaf when mixed with a little water. How numerous are the causes which may check this digestive function of the worm--a function of such importance to a creature which in the space of one month passes from the weight of half a milligramme to that of five, six, seven, or even eight grammes! Pasteur proved that whenever a worm was attacked with _flacherie_, it always had, associated with the food in its intestinal canal, one or other of the microscopic organisms which are invariably to be met with among crushed mulberry leaves. Summing up in a kind of aphorism a series of observations, Pasteur observes: 'Every _ver flat_ is one which digests badly, and, conversely, every worm which digests badly is doomed to perish of _flacherie_, or to furnish a chrysalis and a moth the life of which, through the injury produced by organised ferments, is not normally perfected.' Thus, as in the case of _pebrine_, the morbid symptoms of _flacherie_ are very variable. All depends on the intensity of the evil--that is to say, on the abundance and the nature of the parasites developed in the intestinal canal, and also on the period in the life of the worm when this fermentation begins to show itself. The most dangerous of all these ferments are those of the family of vibrios. If they exist in the first phases of the life of the worm, it dies quickly and very soon becomes putrid, sometimes resolving itself into an infected pus. The disease often manifests itself in a manner particularly distressing and disastrous to the cultivator. The worms have presented the most beautiful appearance up to the time of climbing the heather. The mortality has scarcely been two or three per cent., which is nothing; the moultings have been effected in a perfect manner, when suddenly, some days after the fourth moulting, the worms become languid, crawling with difficulty, and hesitating to take the leaves which are thrown upon their hurdles. If some few have mounted on to the heather, they stretch themselves on the twigs, their bodies swollen with food which they cannot digest. Sometimes they remain there motionless till they die, or, falling, remain suspended only by their false feet. The few moths which have succeeded in piercing their cocoons do not show any corpuscles. They can produce eggs, but these eggs, coming from parents weakened by disease, give rise the following year to a generation threatened with _flacherie_. It is in this sense that the disease may be regarded as hereditary, although the parasites of the intestinal canal to which _flacherie_ is due do not transmit themselves to the eggs or to the worms which issue from them. The worms inherit weakly constitutions, and, being without power of resistance against anything that can derange their digestive functions, they are at the mercy of the accidents of their culture. Too large an assemblage of worms in one nursery; too high a temperature at the time of moulting; a thunderous atmosphere, which predisposes organic matter to fermentation; the use of heated or wet leaves, especially if the wetting be caused by a fog or by the morning or evening dew, which deposits on the leaf the germs suspended in a great mass of air;--these are so many causes calculated to diminish the activity of the digestive functions of the worms, and to produce in consequence a fermentation of the leaf in the intestinal canal--the malady now under consideration. Often also _flacherie_ depends upon mistakes committed by the husbandman while tending his precious 'kine,' to use an expression of the sixteenth century. A Chinese book published on the rearing of silkworms contains a series of little practical counsels. 'The person who takes care of the silkworms,' says this guide to the perfect cultivator, 'ought to wear a simple garment, not lined. He must regulate the temperature of the spinning-house according to the sensation of heat or cold which he experiences; if he feels cold, he may conclude that the worms are cold, and he will increase the fire; if he feels hot, he will conclude that the worms are hot, and he will suitably diminish the fire.' One point which had been ignored before the experiments of Pasteur was the contagious character of _flacherie_. This contagion may surpass that of _pebrine_ itself as regards duration. In _pebrine_ the dried corpusculous matter loses all virulence after the lapse of some weeks. The disease cannot, therefore, communicate itself from one year to another by the corpusculous dust of a rearing establishment. The germs, on the contrary, of the microscopic organisms which provoke fermentation in the mulberry leaves, especially the vibrios, retain their vitality for several years. The dust of a silkworm nursery infected by _flacherie_ appears under a microscope quite full of cysts or spores of vibrios. These spores or cysts rest, like the sleeping beauty in the forest, until a drop of water falls upon them and awakens them into life. Deposited on the leaves which are to serve as nourishment, these germs of vibrios are carried into the intestinal canal of the worm, develop and multiply themselves, and completely disturb the digestive functions, unless the digestion is so strong that the germs are immediately arrested, and disposed of like the food itself. This is what happens when the worms are in full vigour. It is a struggle for life, in which the worms often gain the victory. Giving to some very healthy worms a meal of leaves covered with the dry dust of a silkworm nursery, infected the year before by _pebrine_ and _flacherie_, Pasteur reproduced _flacherie_, and not _pebrine_. Still more readily did he produce the first of these maladies, when he gave, as food, leaves polluted by the contents of the intestinal canal of worms which had died of the disease. As in the case of _pebrine_, the excreta of the worms attacked by _flacherie_, defiling the leaves, carry the mischief to the healthy worms, or add to the dangerous fermentation in the intestines of those which are already in part attacked. To preserve silkworms from accidental _flacherie_, hygienic precautions are sufficient. As regards hereditary _flacherie_, or, to speak more correctly, that which develops itself easily on any diminution of vigour in the eggs and in the embryo, Pasteur again found a remedy by having recourse to the microscope. By means of the microscope it is possible to obtain information as to the health of the worms, the chrysalides, and the moths destined to produce the eggs. Every attention should be directed to the complete exclusion of ferments from the intestinal canal of the worms, and from the stomach-pouch of the chrysalides--a little pouch to which the intestinal canal of the worm is reduced, with its contents more or less transformed. But if there is not time to make this examination for parasitic ferments with the microscope, a simple inspection of the worms in their last stage will suffice. Pasteur laid great stress upon the observation of the worms when they climbed on to the heather. 'If I were a cultivator of silkworms,' he wrote in his beautiful work on the diseases of silkworms, 'I would never hatch an egg produced from worms that I had not observed many times during the last days of their life, so as to make sure of their vigour at the moment when they spin their silk. If you use eggs produced by moths the worms of which have mounted the heather with agility, have shown no signs of _flacherie_ between the fourth moulting and mounting time, and do not contain the least corpuscle of _pebrine_, then you will succeed in all your cultivations.'                                  III. We have now arrived at the end of this long investigation. All the obscurity which enveloped the origin of the diseases of silkworms had now been dispelled. Pasteur had arrived at such accurate knowledge both of the causes of the evil and their different manifestations, that he was able to produce at will either _pebrine_ or _flacherie_. He could so regulate the intensity of the disease as to cause it to appear on a given day, almost at a given hour. He had now to carry into practice the results of his laboratory labours. Since the beginning of the plague, and after some doubts which were soon dispelled, it was clearly seen that all the mischief was to be attributed to the bad condition of the eggs. The remedy of distant explorations for procuring non-infected eggs was both insufficient and precarious. It simply amounted to going very far to seek, and paying very dear, for seed which could not be relied on with certainty. The prosperity of the silkworm culture could only be secured by measures capable of restoring to the native eggs their pristine qualities. The results obtained by Pasteur were sufficient to solve this problem. The struggle against _flacherie_ was easy, but there remained the struggle against _pebrine_. To triumph over this disease, which was so threatening, Pasteur devised a series of observations as simple as they were ingenious. Here is a crop which has perfectly succeeded. The moultings, and the climbing upon the heather, are all that could be desired. The cocoons are finished, and the appearance of the moths alone is waited for. They arrive, and they pair. Then begins the work of the cultivator, who is careful about the production of his eggs. He separates the couples at the end of the day; laying each female moth by itself on a little linen cloth suspended horizontally. The females lay their eggs. After the laying, he takes each female in turn and secures her by a pin passed through the wings to a folded corner of the little cloth, where are grouped some hundreds of eggs which she has laid. The male moth also might be pinned in another corner of the cloth, but the examination of the male is useless, as it has been found that he does not communicate the _pebrine_. The female moth, after having been desiccated by free contact with the air, is examined at leisure, it may be even in the autumn or winter. Nothing is easier than to ascertain whether there are any corpuscles in its dead body. The moth is crushed in a mortar and mixed with a little water, and then a drop of the mixture is examined by the microscope. If corpuscles be found, the bit of cloth corresponding to the examined moth is known, and it is burnt with all the eggs it contains. This method of procuring pure eggs is, in fact, only the rational development of the first inductions which Pasteur had submitted to the Agricultural Committee of Alais in June 1865. At that time he hardly ventured to hope that he should be able to find the means of preparing more than very small quantities of healthy eggs for his experiments; but events were so ordered that the starting-point, which seemed to be purely scientific, unfolded a method susceptible of a widespread practical application. This process of procuring sound eggs is now universally adopted. In the Basses-Alpes, in Ardeche, in Gard, in the Drome, and in other countries, may be met with everywhere, at the time of the cultivation, workshops where hundreds of women and young girls are occupied, with a remarkable division of labour and under the strictest supervision of skilful overseers, in pounding the moths, in examining them microscopically, and in sorting and classifying the little cloths upon which the eggs are deposited.                    *       *       *       *       * But if Pasteur had brought back wealth to ruined countries, if he had returned to Paris happy in the victory he had gained, he had also undergone such fatigues, and had so overstrained himself in the use of the microscope while absorbed in his daily and varied experiments, that in October 1868 he was struck with paralysis of one side. Seeing, as he thought, death approaching, he dictated to his wife a last note on the studies which he had so much at heart. This note was communicated to the Academy of Sciences eight days after this terrible trial. A soul like his, possessing so great a mastery over the body, ended by triumphing over the affliction. Paralysed on the left side, Pasteur never recovered the use of his limbs. To this day, sixteen years after the attack, he limps like a wounded man. But what stages had this wounded man yet to travel over, what triumphs were yet in store for him!                         _DECISIVE EXPERIMENTS._ After having dictated this scientific note, which he thought would have been his last, his courage forsook him for a time. 'I regret to die,' he said to his friend, Sainte-Claire Deville, who had hastened to his bedside; 'I should have wished to render more service to my country.' His life was spared, but for several months Pasteur remained entirely paralysed, incapable of the slightest movement. Smitten thus in his full strength at the age of forty-five, he took a sad review of his own state. Even at the height of his attack his mind had always retained its clearness. He had pointed out to the doctor without any faltering of voice the progressive symptoms of the paralysis. Then reproaching himself for having added to the grief of his wife by thus dwelling on the details of his illness, he never allowed another word to escape him about his infirm condition. Sometimes, even when he heard his two assistants, M. Gernez and M. Duclaux, whose devotion to him during those sad days could only be compared to that of his wife, talking to him of future labours, he entered into these thoughts and appeared to add faith to their hopes. He finished by sharing them. In January 1869, although it was still impossible for him to drag himself about his room, he was so much excited by the contradictions that his system of culture had aroused that he wished to start again for Alais. 'Aided by the method of artificial cultivation,' he remarked, 'we shall soon annihilate these latest oppositions. There is here both a scientific principle and an element of national wealth.' His wish could not be opposed, but a terrible and anxious journey it was! At some leagues from Alais, at a place called Saint Hippolyte-du-Fort, where the earliest experiments were made, Pasteur stopped. He installed himself--we might rather say he encamped--with his family and his assistants, in a more than humble lodging, one of those miserable, cold, paved houses of the rural districts. Seated in his arm-chair, Pasteur directed the experiments, and verified the observations which he had made the year before. Each of his predictions as to the destiny of the different groups of worms was fulfilled to the smallest detail. In the following spring he left for Alais, where he followed in all their phases, from the egg up to the cocoon, the cultivations there undertaken, and he had the happiness of proving once more the certainty of his method. But opposition still continued. The French Government, shaken by the violence and tenacity of the opponents, hesitated to decide upon this process of culture. The Emperor interposed; he instructed Marshal Vaillant to propose to Pasteur to go into Austria to the Villa Vicentina, which belonged to the Prince Imperial. For ten years the silk harvest at this place had not sufficed to pay the cost of eggs. Pasteur accepted with joy the prospect of a great decisive experiment. He traversed France and Italy, reclining in a railway carriage or in an arm-chair, and at last arrived at the Imperial villa near Trieste. Pasteur succeeded in a marvellous manner. The sale of the cocoons gave to the villa a net profit of twenty-six million francs. The Emperor, impressed with the practical value of the system, nominated Pasteur a Senator, in the month of July 1870. But this nomination, like so many other things, was swept away before it had time to appear in the 'Journal Officiel.' Pasteur, however, cared little for the title of Senator. He returned to France on the eve of the declaration of war. A patriot to the heart's core, he learned with poignant grief the news of his country's disasters. The bulletins of defeat, which succeeded each other with mournful monotony, threw him into deep despair. For the first time in his life he had not the strength to work. He lived at his little house in Arbois as one completely vanquished. Those who went into his room found him often bathed in tears. On January 18, 1871, he wrote, to the Dean of the Academy of Medicine at the University of Bonn, a letter in which all his grief and all his pride as a Frenchman were displayed, requesting him to withdraw the diploma of German doctor which the Faculty of Medicine of the University had conferred upon him in 1868. Whilst he wrote this letter, which was a cry of patriotism, his son, enrolled as a volunteer, though hardly eighteen years of age, was gallantly doing his duty in the Army of the East.                           _STUDIES ON BEER._ The war was over. Little by little the life of the country was resumed, and with returning hope the desire and necessity for renewed work. After two years of infirmity, Pasteur at length began to feel the recovery of health. It was like a slow and gentle renewal of all things. He wished to return as soon as possible to his laboratory in Paris to put into execution projects of experiments which had long been working in his brain. At the moment when he was preparing to start, the rebellion of the Commune broke out. M. Duclaux, who had become Professor of the Faculty of Sciences in Clermont-Ferrand, offered the use of his laboratory to his old master. Pasteur accepted it. Eager to commence an investigation which would bring him again to the study of fermentation, he attacked the diseases of beer. But it was not only for the purpose of creating a new link between these researches and his former ones that he occupied himself with this subject, he was also influenced by a somewhat patriotic idea. He dreamt of success in an industry in which Germany is superior to France. He hoped by means of scientific principles, by which commerce would largely profit, to succeed in making for French beer a reputation equal, if not superior, to that of Germany.                    *       *       *       *       * Beer is much more liable to contract diseases than wine. It may be said that while old wine is often to be found, there is no such thing as old beer. It is consumed as fast as it is made. Less acid and less alcoholic than wine, beer is more laden with gummy and saccharine matters, which expose it to rapid changes. Thus the trade in this beverage is constantly struggling with the difficulties of its preservation. The manufacture of beer is simple. It is extracted from germinated barley, or malt, an infusion of which is made and gradually heated to the boiling point. It is then flavoured by hops. When the infusion of malt and hops, which is called 'wort,' is completed, it is subjected to a cooling process, and drawn off into tuns and barrels. It is then that alcoholic fermentation sets in. The cooling ought to be performed rapidly. While the wort is at a high temperature there is nothing to fear, it remains sound; but under 70° Centigrade, and particularly between 25° and 35°, it is easily attacked by injurious ferments--acetic, lactic, or butyric. After the wort is cooled, a little of the yeast proceeding from a former fermentation is added to it, in order that the whole mass of the wort should be invaded as soon as possible after its cooling by the alcoholic ferment alone--the only one, properly speaking, which can produce beer. If this wort were treated in the same way as the must of the grape, if it were abandoned to fermentation without yeast--to so-called spontaneous fermentation--this would hardly ever be purely alcoholic, as in the must of grapes, which is protected by its acidity. Most frequently, instead of beer, an acid or putrid liquid would be obtained. Divers fermentations would simultaneously take place in it. When the wort has fermented and the beer is made, there is still the fear of its rapid deterioration, which necessitates its being quickly consumed. This condition is sometimes disastrous to those employed in the beer trade; and the improvements in the manufacture of beer which have been made during the last forty years have all had for their object the removal of this necessity for the daily production, so to speak, of an article of which the consumption is liable to constant variations. Formerly only one kind of beer was known, the beer of high fermentation. The wort, after having undergone cooling in the troughs, is collected in a large open vat at a temperature of 20°, and yeast is added to it. When the fermentation begins to show itself on the surface of the liquid, by the formation of a light white froth, the wort is transferred to a series of small barrels, which are placed in cellars or store-rooms, kept at a temperature of from 18° to 20° Centigrade. The activity of the fermentation soon causes a foam to rise, which becomes more and more thick and viscous. This is owing to the abundance of yeast which it contains. This yeast, collected in a large trough placed under the casks, is gathered up for future operations. The fermentation lasts for three or four days, then the beer is made and has become clear; the bungs are fixed in the barrels, and they are sent direct to the retail dealer or to the consumer. During the transit, a certain quantity of yeast, fallen to the bottom of the casks, thickens the beer, but a few days of repose suffice to make it again clear and fit to drink, or to be bottled. This system of 'high' fermentation (so called because it begins at a temperature of 18° to 20°, and is raised one or two degrees higher by the act of fermentation itself) is very commonly practised in the north of France, and to a greater extent in the breweries of England. Ale, pale ale, bitter beer, are all beers from high fermentation. The 'low' fermentation, which is almost exclusively employed in Germany, and which is spreading more and more in France, consists in a slow fermentation, at low temperature, during which the yeast settles at the bottom of the tubs and casks. The wort, after it has been cooled, is passed into open wooden tuns, and the working of the yeast takes place at a temperature of about 6° Centigrade. This temperature is maintained by means of floats, in the form of cones or cylinders, thrown into the fermenting tuns and kept filled with ice. The fermentation lasts for ten, fifteen, and even twenty days. When the beer is drawn off, the yeast is collected from the bottom of the fermenting tuns. This kind of beer, which is sometimes called German beer, sometimes Strasburg beer, is generally much more esteemed than the other, but it requires certain expensive, or at least inconvenient, conditions. There must be ice-caves, where the temperature is maintained all the year round at a few degrees only above zero. This makes it necessary to have enormous piles of ice. It has been calculated that for one single hectolitre of good beer, from the beginning of the cooling of the wort until the time when it is fit for sale, 100 kilogrammes of ice are required. The 'low' beer, called also _biere de garde_, beer for keeping, is principally manufactured in winter, and is preserved in ice-caves until the summer. It is not only the taste of the consumers which has favoured the manufacture of beer of low fermentation everywhere except in England; it is also the advantage this beer possesses in being much less liable to deterioration than the other. By employing ice, the brewer may manufacture in winter, or in the beginning of spring, and thus place himself in a position to meet the demands of consumption without fear of seeing his beer attacked by disease.                    *       *       *       *       * All the diseases of beer, as Pasteur has shown, are caused exclusively by the development of little microscopic fungi, or organised ferments, the germs of which are brought by the dust constantly floating in the air, or which gets mixed with the original substances used in the manufacture. 'By the expression _diseases_ of wort and of beer, I mean,' said Pasteur, 'those serious alterations which affect the quality of these liquids so as to render them disagreeable to the taste, especially when they have been kept for some time, and which cause the beer to be described as sharp, sourish, turned, ropy, putrid.' The wort of beer, after it has been raised to the boiling heat, may, as Pasteur's experiments testify, be preserved indefinitely, even in the highest atmospheric temperatures, when in contact with air free from the germs of the lower microscopic organisms. The must, leavened by the addition of pure yeast, kept free from foreign organisms, contains nothing but the alcoholic ferment, and undergoes no other changes than those due to the action of the oxygen, which does not give rise to acidity, putridity, or bitterness. Since the causes of deterioration are the same in beer as in wine, would it not appear as if the action of heat must be the best preservative? But beer is a drink necessarily charged with carbonic acid, and the application of heat to considerable masses of the liquid would expel this gas. It would be a very complicated business to attempt to preserve this gas, or to introduce it afresh after it had been expelled. This difficulty does not arise when the beer is bottled. At a temperature of 50° to 55°, the process of heating not only cannot take away from the beer all its carbonic acid, but it does not prevent the secondary fermentation from taking place to a certain extent, and this allows of the beer being heated immediately after it is put into bottles. This heating of the beer is practised on a large scale in Europe and in America. In honour of Pasteur the process is called _Pasteurisation_, and the beer_Pasteurised_ beer.