Antiseptics During World War I, Fleming with
Leonard Colebrook and Sir Almroth Wright joined the war efforts and practically moved the entire Inoculation Department of St Mary's to the British military hospital at
Boulogne-sur-Mer. Serving as a temporary lieutenant of the Royal Army Medical Corps, he witnessed the death of many soldiers from
sepsis resulting from infected
wounds.
Antiseptics, which were used at the time to treat infected wounds, he observed, often worsened the injuries. In a 1917 article published in the medical journal
The Lancet, he described an ingenious experiment, which he was able to conduct as a result of his own
glassblowing skills, in which he explained why antiseptics were killing more soldiers than infection itself during the war. Antiseptics worked well on the surface, but deep wounds tended to shelter
anaerobic bacteria from the antiseptic agent, and antiseptics seemed to remove beneficial agents produced that protected the patients in these cases at least as well as they removed bacteria, and did nothing to remove the bacteria that were out of reach. Wright strongly supported Fleming's findings, but despite this, most army physicians over the course of the war continued to use antiseptics even in cases where this worsened the condition of the patients.
Discovery of lysozyme At St Mary's Hospital, Fleming continued his investigations into bacteria culture and antibacterial substances. As his research scholar at the time V. D. Allison recalled, Fleming was not a tidy researcher and usually expected unusual bacterial growths in his culture plates. Fleming had teased Allison of his "excessive tidiness in the laboratory", and Allison rightly attributed such untidiness as the success of Fleming's experiments, and said, "[If] he had been as tidy as he thought I was, he would not have made his two great discoveries." In late 1921, while Fleming was maintaining
agar plates for bacteria, he found that one of the plates was contaminated with bacteria from the air. When he added nasal mucus, he found that the mucus inhibited the bacterial growth. Surrounding the mucus area was a clear transparent circle (1 cm from the mucus), indicating the
killing zone of bacteria, followed by a glassy and translucent ring beyond which was an opaque area indicating normal bacterial growth. In the next test, he used bacteria maintained in saline that formed a yellow suspension. Within two minutes of adding fresh mucus, the yellow saline turned completely clear. Although he was able to obtain larger amounts of lysozyme from egg whites, the enzyme was only effective against small counts of harmless bacteria, and therefore had little therapeutic potential. This indicates one of the major differences between
pathogenic and harmless bacteria. The species was reassigned as
Micrococcus luteus in 1972. The "Fleming strain" (NCTC2665) of this bacterium has become a model in different biological studies. The importance of lysozyme was not recognised, and Fleming was well aware of this, in his presidential address at the
Royal Society of Medicine meeting on 18 October 1932, he said:I choose lysozyme as the subject for this address for two reasons, firstly because I have a fatherly interest in the name, and, secondly, because its importance in connection with natural immunity does not seem to be generally appreciated. In his Nobel lecture on 11 December 1945, he briefly mentioned lysozyme, saying, "Penicillin was not the first antibiotic I happened to discover." It was only towards the end of the 20th century that the true importance of Fleming's discovery in immunology was realised as lysozyme became the first
antimicrobial protein discovered that constitute part of our
innate immunity.
Discovery of penicillin Experiment By 1927, Fleming had been investigating the properties of
staphylococci. He was already well known from his earlier work, and had developed a reputation as a brilliant researcher. In 1928, he studied the variation of
Staphylococcus aureus grown under natural condition, after the work of Joseph Warwick Bigger, who discovered that the bacterium could grow into a variety of types (strains). On 3 September 1928, Fleming returned to his laboratory having spent a holiday with his family at Suffolk. Before leaving for his holiday, he inoculated staphylococci on culture plates and left them on a bench in a corner of his laboratory. On his return, Fleming noticed that one culture was contaminated with a fungus, and that the colonies of staphylococci immediately surrounding the fungus had been destroyed, whereas other staphylococci colonies farther away were normal, famously remarking "That's funny". Fleming showed the contaminated culture to his former assistant Merlin Pryce, who reminded him, "That's how you discovered lysozyme." He identified the mould as being from the genus
Penicillium. He suspected it to be
P. chrysogenum, but a colleague Charles J. La Touche identified it as
P. rubrum. It was later corrected as
P. notatum and then officially accepted as
P. chrysogenum. In 2011, it was resolved as
P. rubens. The laboratory in which Fleming discovered and tested penicillin is preserved as the
Alexander Fleming Laboratory Museum in St. Mary's Hospital,
Paddington. The source of the fungal contaminant was established in 1966 as coming from La Touche's room, which was directly below Fleming's. Fleming grew the mould in a pure culture and found that the culture broth contained an antibacterial substance. He investigated its anti-bacterial effect on many organisms, and noticed that it affected bacteria such as staphylococci and many other
Gram-positive pathogens that cause
scarlet fever,
pneumonia,
meningitis and
diphtheria, but not
typhoid fever or
paratyphoid fever, which are caused by
Gram-negative bacteria, for which he was seeking a cure at the time. It also affected
Neisseria gonorrhoeae, which causes
gonorrhoea, although this bacterium is Gram-negative. After some months of calling it "mould juice" or "the inhibitor", he gave the name
penicillin on 7 March 1929 for the antibacterial substance present in the mould. but little attention was paid to the article. His problem was the difficulty of producing penicillin in large amounts, and moreover, isolation of the main compound. Even with the help of Harold Raistrick and his team of biochemists at the
London School of Hygiene & Tropical Medicine, chemical purification was futile. "As a result, penicillin languished largely forgotten in the 1930s", as
Milton Wainwright described. no one believed him. As Allison, his companion in both the Medical Research Club and international congress meeting, remarked the two occasions:[Fleming at the Medical Research Club meeting] suggested the possible value of penicillin for the treatment of infection in man. Again there was a total lack of interest and no discussion. Fleming was keenly disappointed, but worse was to follow. He read a paper on his work on penicillin at a meeting of the International Congress of Microbiology, attended by the foremost bacteriologists from all over the world. There was no support for his views on its possible future value for the prevention and treatment of human infections and discussion was minimal. Fleming bore these disappointments stoically, but they did not alter his views or deter him from continuing his investigation of penicillin. Shortly after the team published its first results in 1940, Fleming telephoned
Howard Florey, Chain's head of department, to say that he would be visiting within the next few days. When Chain heard that Fleming was coming, he remarked "Good God! I thought he was dead."
Norman Heatley suggested transferring the active ingredient of penicillin back into water by changing its acidity. This produced enough of the drug to begin testing on animals. There were many more people involved in the Oxford team, and at one point the entire
Sir William Dunn School of Pathology was involved in its production. After the team had developed a method of purifying penicillin to an effective first stable form in 1940, several clinical trials ensued, and their amazing success inspired the team to develop methods for mass production and mass distribution in 1945. Fleming was modest about his part in the development of penicillin, describing his fame as the "Fleming Myth" and he praised Florey and Chain for transforming the laboratory curiosity into a practical drug. Fleming was the first to discover the properties of the active substance, giving him the privilege of naming it: penicillin. He also kept, grew, and distributed the original mould for twelve years, and continued until 1940 to try to get help from any chemist who had enough skill to make penicillin. Sir
Henry Harris summed up the process in 1998 as: "Without Fleming, no Chain; without Chain, no Florey; without Florey, no Heatley; without Heatley, no penicillin." The discovery of penicillin and its subsequent development as a prescription drug mark the start of modern
antibiotics.
Medical use and mass production In his first clinical trial, Fleming treated his research scholar Stuart Craddock who had developed severe infection of the
nasal antrum (
sinusitis). The treatment started on 9 January 1929 but without any effect. It probably was due to the fact that the infection was with influenza bacillus (
Haemophilus influenzae), the bacterium which he had found unsusceptible to penicillin. Although Wright reportedly said that it "seemed to work satisfactorily", there are no records of its specific use. Cecil George Paine, a pathologist at the
Royal Infirmary in Sheffield and former student of Fleming, was the first to use penicillin successfully for medical treatment. He cured eye infections (
conjunctivitis) of one adult and three infants (
neonatal conjunctivitis) on 25 November 1930. Fleming also successfully treated severe conjunctivitis in 1932. Keith Bernard Rogers, who had joined St Mary's as medical student in 1929, was captain of the London University rifle team and was about to participate in an inter-hospital rifle shooting competition when he developed conjunctivitis. Fleming applied his penicillin and cured Rogers before the competition. It is said that the "penicillin worked and the match was won." However, the report that "Keith was probably the first patient to be treated clinically with penicillin ointment" is no longer true as Paine's medical records showed up. There is a popular assertion both in popular and scientific literature that Fleming largely abandoned penicillin work in the early 1930s. In his review of
André Maurois's
The Life of Sir Alexander Fleming, Discoverer of Penicillin, William L. Kissick went so far as to say that "Fleming had abandoned penicillin in 1932... Although the recipient of many honors and the author of much scientific work, Sir Alexander Fleming does not appear to be an ideal subject for a biography." This is false, as Fleming continued to pursue penicillin research. As late as in 1939, Fleming's notebook shows attempts to make better penicillin production using different media. As to the chemical isolation and purification,
Howard Florey and
Ernst Chain at the
Radcliffe Infirmary in Oxford took up the research to mass-produce it, which they achieved with support from World War II military projects under the British and US governments. By mid-1942, the Oxford team produced the pure penicillin compound as yellow powder. In August 1942, Harry Lambert (an associate of Fleming's brother Robert) was admitted to St Mary's Hospital due to a life-threatening infection of the nervous system (streptococcal
meningitis). Fleming published the clinical case in
The Lancet in 1943. Upon this medical breakthrough, Allison informed the British
Ministry of Health of the importance of penicillin and the need for mass production. The
War Cabinet was convinced of the usefulness upon which Sir
Cecil Weir, Director General of Equipment, called for a meeting on the mode of action on 28 September 1942. The Penicillin Committee was created on 5 April 1943. The committee consisted of Weir as chairman, Fleming, Florey, Sir
Percival Hartley, Allison and representatives from pharmaceutical companies as members. The main goals were to produce penicillin rapidly in large quantities with collaboration of American companies, and to supply the drug exclusively for
Allied armed forces.
Antibiotic resistance Fleming also discovered very early that bacteria developed
antibiotic resistance whenever too little penicillin was used or when it was used for too short a period. Almroth Wright had predicted antibiotic resistance even before it was noticed during experiments. Fleming cautioned about the use of penicillin in his many speeches around the world. On 26 June 1945, he made the following cautionary statements: "the microbes are educated to resist penicillin and a host of penicillin-fast organisms is bred out ... In such cases the thoughtless person playing with penicillin is morally responsible for the death of the man who finally succumbs to infection with the penicillin-resistant organism. I hope this evil can be averted." He cautioned not to use penicillin unless there was a properly diagnosed reason for it to be used, and that if it were used, never to use too little, or for too short a period, since these are the circumstances under which bacterial resistance to antibiotics develops. It had been experimentally shown in 1942 that
S. aureus could develop penicillin resistance under prolonged exposure. Elaborating the possibility of penicillin resistance in clinical conditions in his Nobel Lecture, Fleming said:The time may come when penicillin can be bought by anyone in the shops. Then there is the danger that the ignorant man may easily underdose himself and by exposing his microbes to non-lethal quantities of the drug make them resistant. ==Personal life==