Molecular asymmetry shapes from each other to form two piles of crystals: in solution one form rotated light to the left, the other to the right, while an
equal mixture of the two forms canceled each other's effect, and does not rotate the
polarized light. In Pasteur's early work as a
chemist, beginning at the
École Normale Supérieure, and continuing at Strasbourg and Lille, he examined the chemical, optical and crystallographic properties of a group of compounds known as
tartrates. A solution of this compound derived from living things
rotated the
plane of polarization of light passing through it. The problem was that tartaric acid derived by
chemical synthesis had no such effect, even though its chemical reactions were identical and its elemental composition was the same. The (2
R,3
R)- and (2
S,3
S)- tartrates were isometric, non-superposable mirror images of each other. This was the first time anyone had demonstrated
molecular chirality, and also the first explanation of
isomerism. A memoire was subsequently published on 30 November 1857. In the memoir, he developed his ideas stating that: "I intend to establish that, just as there is an alcoholic ferment, the yeast of beer, which is found everywhere that sugar is decomposed into alcohol and carbonic acid, so also there is a particular ferment, a
lactic yeast, always present when
sugar becomes lactic acid." This memoir on alcoholic fermentation was published in full form in 1858.
Jöns Jacob Berzelius and
Justus von Liebig had proposed the theory that fermentation was caused by decomposition. Pasteur demonstrated that this theory was incorrect, and that yeast was responsible for fermentation to produce alcohol from sugar. He also demonstrated that, when a different microorganism contaminated the wine, lactic acid was produced, making the wine sour. Pasteur's research also showed that the growth of micro-organisms was responsible for spoiling beverages, such as beer, wine and milk. With this established, he invented a process in which liquids such as milk were heated to a temperature between 60 and 100 °C. Pasteur patented the process, to fight the "diseases" of wine, in 1865. The method became known as
pasteurization, and was soon applied to beer and milk. Beverage contamination led Pasteur to the idea that micro-organisms infecting animals and humans cause disease. He proposed preventing the entry of micro-organisms into the human body, leading
Joseph Lister to develop
antiseptic methods in surgery. In 1866, Pasteur published
Études sur le Vin, about the diseases of wine, and he published
Études sur la Bière in 1876, concerning the diseases of beer. Silkworms with pébrine were covered in corpuscles. In the first three years, Pasteur thought that the corpuscles were a symptom of the disease. In 1870, he concluded that the corpuscles were the cause of pébrine (it is now known that the cause is a
microsporidian). Pasteur also showed that the disease was hereditary. Pasteur developed a system to prevent pébrine: after the female moths laid their eggs, the moths were turned into a pulp. The pulp was examined with a microscope, and if corpuscles were observed, the eggs were destroyed.
Spontaneous generation ) used by Pasteur Following his fermentation experiments, Pasteur demonstrated that the skin of grapes was the natural source of yeasts, and that sterilized grapes and grape juice never fermented. He drew grape juice from under the skin with sterilized needles, and also covered grapes with sterilized cloth. Both experiments could not produce wine in sterilized containers. Pouchet stated that air everywhere could cause spontaneous generation of living organisms in liquids. In the late 1850s, he performed experiments and claimed that they were evidence of spontaneous generation. Pasteur also used
swan neck flasks containing a fermentable liquid. Air was allowed to enter the flask via a long curving tube that made dust particles stick to it. Nothing grew in the broths unless the flasks were tilted, making the liquid touch the contaminated walls of the neck. This showed that the living organisms that grew in such broths came from outside, on dust, rather than spontaneously generating within the liquid or from the action of pure air. These were some of the most important experiments disproving the theory of spontaneous generation. Pasteur gave a series of five presentations of his findings before the French Academy of Sciences in 1881, which were published in 1882 as
Mémoire ''Sur les corpuscules organisés qui existent dans l'atmosphère: Examen de la doctrine des générations spontanées
(Account of Organized Corpuscles Existing in the Atmosphere: Examining the Doctrine of Spontaneous Generation''). Pasteur won the Alhumbert Prize in 1862. He concluded that:
Silkworm disease In 1865,
Jean-Baptiste Dumas, chemist, senator and former Minister of Agriculture and Commerce, asked Pasteur to study a new disease that was decimating
silkworm farms from the south of France and Europe, the
pébrine, characterized on a macroscopic scale by black spots and on a microscopic scale by the "
Cornalia corpuscles". Pasteur accepted and made five long stays in
Alès, between 7 June 1865 and 1869.
Initial errors Arriving in Alès, Pasteur familiarized himself with pébrine and also with another disease of the silkworm, known earlier than pebrine:
flacherie or dead-flat disease. Contrary, for example, to
Quatrefages, who coined the new word
pébrine, Pasteur made the mistake of believing that the two diseases were the same and even that most of the diseases of silkworms known up to that time were identical with each other and with pébrine. It was in letters of 30 April and 21 May 1867 to Dumas that he first made the distinction between pébrine and flacherie. He made another mistake: he began by denying the "parasitic" (microbial) nature of pébrine, which several scholars (notably
Antoine Béchamp) considered well established. Even a note published on 27 August 1866 by
Balbiani, which Pasteur at first seemed to welcome favourably had no effect, at least immediately. "Pasteur is mistaken. He would only change his mind in the course of 1867".
Victory over pébrine At a time when Pasteur had not yet understood the cause of the pébrine, he propagated an effective process to stop infections: a sample of chrysalises was chosen, they were crushed and the corpuscles were searched for in the crushed material; if the proportion of corpuscular pupae in the sample was very low, the chamber was considered good for reproduction. This method of sorting "seeds" (eggs) is close to a method that Osimo had proposed a few years earlier, but whose trials had not been conclusive. By this process, Pasteur curbed pébrine and saved much of the silk industry in the Cévennes.
Flacherie resists In 1878, at the
Congrès international séricicole, Pasteur admitted that "if pébrine is overcome, flacherie still exerts its ravages". He attributed the persistence of flacherie to the fact that the farmers had not followed his advice. In 1884,
Balbiani, who disregarded the theoretical value of Pasteur's work on silkworm diseases, acknowledged that his practical process had remedied the ravages of pébrine, but added that this result tended to be counterbalanced by the development of flacherie, which was less well known and more difficult to prevent. Despite Pasteur's success against pébrine, French sericulture had not been saved from damage. (See
:fr:Sériciculture in the French Wikipedia.)
Immunology and vaccination Chicken cholera Pasteur's first work on vaccine development was on
chicken cholera. He received the bacteria samples (later called
Pasteurella multocida after him) from
Henry Toussaint. Being unable to conduct the experiments himself due to a stroke in 1868, Pasteur relied heavily on his assistants Emile Roux and Charles Chamberland. The work with chicken cholera was initiated in 1877, and by the next year, Roux was able to maintain a stable culture using broths. As documented later by Pasteur in his notebook in March 1880, in October 1879, being delayed in returning to the laboratory due to his daughter's wedding and ill health, he instructed Roux to start a new chicken cholera culture using bacteria from a culture that had sat since July. The two chickens inoculated with this new culture showed some symptoms of infection, but instead of the infections being fatal, as they usually were, the chickens recovered completely. After further incubation of the culture for an additional 8 days, Roux again inoculated the same two chickens. As was also noted by Pasteur in his notebook in March 1880, and contrary to some accounts, this time the chickens died. Thus, although the attenuated bacteria did not provide immunity, these experiments provided important clues as to how bacteria could be artificially attenuated in the laboratory. As a result, upon Pasteur's return to the laboratory, the focus of the research was directed at creating a vaccine through attenuation. In February 1880, Pasteur presented his results to the French Academy of Sciences as "
Sur les maladies virulentes et en particulier sur la maladie appelée vulgairement choléra des poules (On virulent diseases, and in particular on the disease commonly called chicken cholera)" and published it in the academy's journal (''
Comptes-Rendus hebdomadaires des séances de l'Académie des Sciences''). He attributed that the bacteria were weakened by contact with oxygen.
Anthrax Following the results with chicken cholera, Pasteur eventually utilized the immunization method developed for chicken cholera to create a vaccine for
anthrax, which affected
cattle. In 1877, Pasteur had earlier directed his laboratory to culture the bacteria from the blood of infected animals, following the discovery of the bacterium by Robert Koch. Many cattle were dying of anthrax in "cursed fields". Pasteur's interest in creating a vaccine for anthrax was greatly stimulated when on 12 July 1880, Henri Bouley read before the French Academy of Sciences a report from
Henry Toussaint, a
veterinary surgeon, who was not a member of the academy. Toussaint had developed anthrax vaccine by killing the bacilli by heating at 55 °C for 10 minutes. He tested his vaccine on eight dogs and 11 sheep, half of which died after inoculation. It was not a great success. Upon hearing the news, Pasteur immediately wrote to the academy that he could not believe that dead vaccine would work and that Toussaint's claim "overturns all the ideas I had on viruses, vaccines, etc." However, Pasteur's laboratory found that anthrax bacillus was not easily weakened by culturing in air as it formed spores – unlike chicken cholera bacillus. In early 1881, his laboratory discovered that growing anthrax bacilli at about 42 °C made them unable to produce spores, and he described this method in a speech to the French Academy of Sciences on 28 February. On 21 March, despite inconsistent results, he announced successful vaccination of sheep. To this news, veterinarian Hippolyte Rossignol proposed that the Société d'agriculture de Melun organize an experiment to test Pasteur's vaccine. Pasteur signed an agreement accepting the challenge on 28 April. Pasteur's assistants, Roux and Chamberland, who were assigned the task of conducting the trial, were concerned about the unreliability of the attenuated vaccine, and therefore Chamberland secretly prepared an alternative vaccine using chemical inactivation. On 31 May, Roux and Chamberland next injected the animals with the fresh virulent culture of anthrax bacillus. The official result was observed and analyzed on 2 June in the presence of over 200 spectators, with Pasteur himself in attendance. The results were as Pasteur had bravely predicted: "I hypothesized that the six vaccinated cows would not become very ill, while the four unvaccinated cows would perish or at least become very ill." His report to the French Academy of Sciences on 13 June concludes:Pasteur did not directly disclose how he prepared the vaccines used at Pouilly-le-Fort. The notion of a weak form of a disease causing immunity to the virulent version was not new; this had been known for a long time for
smallpox. Inoculation with smallpox (
variolation) was known to result in a much less severe disease, and greatly reduced mortality, in comparison with the naturally acquired disease.
Edward Jenner had also studied
vaccination using
cowpox (
vaccinia) to give cross-immunity to smallpox in the late 1790s, and by the early 1800s vaccination had spread to most of Europe. The difference between smallpox vaccination and
anthrax or
chicken cholera vaccination was that the latter two disease organisms had been artificially weakened, so a naturally weak form of the disease organism did not need to be found. In 1876,
Robert Koch had shown that
Bacillus anthracis caused anthrax. In his papers published between 1878 and 1880, Pasteur only mentioned Koch's work in a footnote. Koch met Pasteur at the Seventh
International Medical Congress in 1881. A few months later, Koch wrote that Pasteur had used impure cultures and made errors. In 1882, Pasteur replied to Koch in a speech, to which Koch responded aggressively. Thuillier identified the bacillus that caused the disease in March 1883.
Rabies ", caricature of Pasteur in the London magazine
Vanity Fair, January 1887 Pasteur's laboratory produced the first vaccine for
rabies using a method developed by his assistant Roux, The rabies vaccine was initially created by
Emile Roux, a French doctor and a colleague of Pasteur, who had produced a killed vaccine using this method. This vaccine was used on 9-year-old
Joseph Meister, on 6 July 1885, after the boy was badly mauled by a rabid dog. Pasteur was hailed as a hero and the legal matter was not pursued. Pasteur began treatment of Jean-Baptiste Jupille on 20 October 1885, and the treatment was successful. Because of his study in germs, Pasteur encouraged doctors to sanitize their hands and equipment before surgery. Prior to this, few doctors or their assistants practiced these procedures.
Ignaz Semmelweis and
Joseph Lister had earlier practiced hand sanitizing in medical contexts in the 1860s. == Controversies ==