It was all a matter of luck. Fleming had decided not to store his culture in a warm incubator (and why would he if he had put it in the washing pile?), and London was then hit by a cold spell, giving the mould a chance to grow. Later, as the temperature rose (London experienced unusually warm weather), the staphylococcus bacteria flourished until it covered the entire plate, except for the area surrounding the mouldy contaminant—the mysterious spore. In short, while Fleming was out of the lab, London experienced ideal conditions for bacteria and mould growth.
So was this Fleming’s ‘Eureka!’ moment, an instant of great personal insight and deductive reasoning? Some biographers see it in this light. Whatever the truth of the matter, Fleming took a sample of the mould and found that it was the somewhat rare Penicillium notatum. His discovery started a chain of events that would literally change the world—but the world had to wait ten years before Florey and his Oxford team got on the case. Fleming’s notes reveal that he did not consider his discovery to be momentous. He merely concluded that the mould was producing an antibiotic substance, which he named penicillin.[9]
Fleming worked on penicillin for four years, but refining it and growing it was a difficult process for him. Every attempt he made to isolate the active ingredient from the broth used to cultivate the mould failed because he lacked the chemical expertise to extract the bacteria-killing substance. Consequently he was unable to test its efficacy against general infections and did not try it against syphilis. In 1929 Fleming published a report describing his initial work on penicillin and its potential uses in the British Journal of Experimental Pathology but it raised little interest.[10] There were other researchers who experienced similar frustration with penicillin in the early 1930s. Fleming had willingly provided them with samples of his mould in order to safeguard the unusual strain of Penicillium notatum. This unselfish act was to prove critical ten years down the track when penicillin finally emerged from obscurity.
By 1932 Fleming had put penicillin aside and was not involved in any way in its later development nor did he discover any other ‘magic bullet’. Fleming would later say that his only merit was that he did not neglect the observation he had made when he found the mould and that he pursued it as a bacteriologist.[11] But while Alexander Fleming did not pursue penicillin any further, he would eventually pursue the glory for discovering it.
BEFORE FLEMING
Three thousand years before penicillin, moulds and fermented materials had been used to cure various skin infections, even though why or how they worked was not understood. Traditional healing practice in many cultures included the application of fungi to wounds or cuts. The Chinese had used mouldy soybeans, the Greeks had used mouldy cheese and the Australian Aboriginals took mouldy bark from the shaded side of trees to make bandages.
It was not until the late 1800s that scientific studies of antibiotics began, made possible after Pasteur had established his Germ Theory of Disease which proved that infectious diseases are spread by micro-organisms. When carrying out his work on anthrax, Pasteur had observed that mould inhibited the growth of anthrax; that when more than one bacterial pathogen is living in the same space, including on human tissue, there is a microscopic struggle for existence. One life form produces an ‘antibiotic’, meaning ‘against life’, to vanquish others.
British surgeon Joseph Lister, a proponent and beneficiary of Pasteur’s work, was aware that bacteria did not grow in urine contaminated with mould but he was unable to identify the active substance in the mould. In 1875, long before Fleming’s observations, John Tyndall, an English physician had discovered that Penicillium killed bacteria. He published his findings in Transaction of the Royal Society, but did no further work on the mould because before Germ Theory was established it was not known that most common infections are caused by bacteria. In 1912 Ernest Duchesne, a French medical student, successfully tested a substance from mould that inhibited bacterial growth in animals.[12] Duchesne died not long after and so never saw the impact of what he had discovered.
In 1925 the Belgian researcher André Gratia, while researching ways to kill bacteria, discovered bacteriocins. These are toxins produced by bacteria to inhibit the growth of similar or closely related bacterial strains. He called the bacteriocin a colicine because it killed E. coli. He was also one of the first to discover bacteriophage, often referred to as phage, which is any virus that attacks bacteria. Smaller than bacteria, a phage infects bacteria by attaching itself to the cell wall of bacterium and injecting genetic material into it.[13] Gratia developed selection techniques that were adopted by other antibiotic hunters. Today there are hospitals in Russia where phage are used to treat infections.
Penicillin G was the first naturally occurring antibiotic to be discovered, the same one that killed bacteria during Duchesne’s work in 1896 and that both Fleming in 1928 and Florey in 1939 experimented with. Today it is the most widely used form of penicillin and is obtained in a number of forms from Penicillium moulds. There are now more than 60 antibiotics, which work by preventing bacteria from reproducing. Bacteria reproduce by dividing to form two new cells. They enlarge to about twice their size before the DNA chromosome is copied. The two new chromosomes move apart and a cell wall forms between them. But penicillin prevents the new cell wall from forming so the bacteria cannot reproduce and the wonderful consequence is that the disease cannot spread.[14] Under a microscope you can see bacteria growing in the presence of penicillin. They look like thin jellybeans and as they extend they eventually rupture because they are unable to divide. Before penicillin they just kept dividing and dividing inside the unfortunates who had been infected.
ENTER HOWARD FLOREY
The scene was set for the development of penicillin in 1935 when Howard Florey, considered an outsider because he was Australian, was appointed Professor of Pathology at Oxford University, the youngest person ever to hold that position. It was a long way from the modest stone cottage in Malvern, a suburb of Adelaide, South Australia, where Florey was born on 24 September 1898. He was the only son of Joseph and Bertha Florey and was doted on by four older sisters. Joseph’s first wife died of tuberculosis soon after they migrated from England in search of a warmer climate and Joseph remarried. An ambitious man, he started a shoe business in Adelaide and quickly established factories across the Australian continent.
By the time Howard was eight, his father was wealthy and the family moved to a sandstone mansion in the Adelaide Hills. Howard was a happy, energetic child who spent time roaming the green fields around his home in much the same way that Alexander Fleming roamed the Scottish countryside, both developing an interest in nature. Brilliant at schoolwork and outstanding at sport, Howard’s commitment to study and his natural talents earned him bursaries which helped to fund his education. When he was thirteen, Howard attended St Peter’s Collegiate School in Adelaide.
Florey later recalled that at about the age of twelve, although he was not very good at mathematics, he was interested in chemistry and already had some idea of doing research. When he told one of his sisters about his ambition she asked him if he wanted to be a ‘sort of Pasteur’.[15] It was the ultimate irony. Florey admitted that he did not know what she meant. And yet, in the not too distant future Howard Florey would advance both the work and the ideals of Louis Pasteur. Fortunately, his interest was nurtured during his school years by an inspired chemistry teacher.
When World War I broke out in 1914 many students from St Peter’s Collegiate School went into the armed forces. Australia, although it had recently become a nation, still had strong colonial bonds and supported the British in their fight against Germany. Howard, who was about to turn eighteen, wanted to join the war effort but his parents were bitterly opposed. It was not a good time for the family. Joseph Florey’s health and his businesses were failing. In an attempt to persuade his father, Howard wrote a letter in which he said that he believed he should enlist:
...not because of a lot of balderdash that’s talked about king a
nd country, and other patriotic rubbish, but because it’s the right thing to do ... When I hear a chap saying he’d like to go to the war, I feel like calling him a liar or a damn fool. I don’t want to go, but I ought to.[16]The Floreys refused their consent despite Howard’s entreaties and so Howard entered medical school at Adelaide University instead of the armed forces. It was here that he met his future wife, Ethel Reed, a fellow medical student. The two were destined to make history together but their marriage would suffer as a result. Ethel was spirited and intelligent, they both had an interest in sport and, according to those who knew them, both were extremely strong-willed.[17]
Mary Ethel Hayter Reed was born in 1900. Ethel’s father was the manager of the Bank of Australasia in Adelaide and her mother was proud of her aristocratic French ancestry. The family home was large and elegant. Called ‘The Red House’, it overlooked a park in North Adelaide. Mr Reed gave a tenth of his income to charity but this generosity did not extend to his family. Although his four children were well educated they were constantly reminded of the cost. In 1919, against her parents’ wishes, Ethel began studying medicine at the University of Adelaide and was the only woman in her year. Ethel was ambitious, a trait that would have been necessary at the time for a woman to achieve success in the degree and career she had chosen.
Friends and fellow students from that time described Ethel variously as popular, pretty, vivacious and fond of parties, dancing and tennis. Because she was one of the few women studying medicine, in January 1920, when Howard Florey was the editor of the Adelaide Medical Students’ Society’s publication, The Review, he wrote to Ethel asking her to contribute an article on ‘Women in Medicine’.[18] Over the next six years Howard wrote 153 letters and Ethel kept every one. In December that year Ethel contracted the first of many bouts of pleurisy and in May 1921, when it was feared she had tuberculosis, she was advised to spend a year in the mountains recuperating.
While Ethel was coping with illness, Howard Florey was sailing to England to take up a Rhodes scholarship at Oxford University. Late in 1921, aged 23, Florey left Australia wondering if he would ever return as there was a dearth of opportunities for researchers in the discipline of physiology.[19] As Florey made the long sea voyage to England as a ship’s doctor, unbeknown to him, the research that Alexander Fleming was engaged in at St Mary’s Hospital would influence the course of his own research and indeed his life. Fleming and Florey would share a Nobel Prize, but would never work together and would never be friends; ironically, however, their individual contributions to the discovery of penicillin would often be confused.
At the time that Florey arrived in England, Australians had a fairly mixed reputation because of the stigma of their colonial background. Florey was the archetypal Australian, somewhat brash and he never wore a collar and tie in the laboratory. There are various descriptions of him as a rough, tough Australian, energetic, tense like a coiled spring and completely uncompromising. By his own account, however, Florey was well treated at Oxford, social faux pas often being excused because he was ‘just one of these rough colonials’.[20] His acceptance had much to do with his outstanding intellect, which earned him enormous respect in the academic sphere.
After completing his Bachelor of Science in the Honours School in Physiology at Oxford Florey came under the attention of Sir Charles Sherrington, whom Florey considered to be one of the greatest physiologists of all time, while doing research in his department. Sherrington, who had been asked to recommend someone for a studentship at Cambridge, approached Florey and enquired if he was interested in experimental pathology. Sherrington believed that if the right person could combine physiology and pathology then science had a lot to gain and that Florey was that person.[21] Florey’s name was put forward and he took up the position at Cambridge where, as was expected, he excelled.
According to biographers, Florey did not make friends easily and was lonely at times but in his correspondence with Ethel in Australia he said that he particularly liked London, that it pulsated with life. He was scathing of the upper classes, whom he said squandered their ‘infinite educational advantages’.[22] The letters went back and forth and in one Howard asked Ethel to join him in England. They had not seen each other for five years.
Ethel had graduated in 1924 as a Bachelor of Medicine and a Bachelor of Surgery and worked as a house surgeon at the Royal Adelaide Hospital where she was highly regarded. But her health problems persisted. In 1926 while Ethel and Howard were debating the possibility of marriage in the exchange of letters, Ethel was becoming increasingly deaf after having developed otosclerosis, an abnormal bone growth in the middle ear that causes hearing loss. The decision to be together was made and in 1927 Ethel completed a six-month residency at Adelaide Children’s Hospital but just before setting sail for England she had another health scare and had some enlarged tuberculous glands surgically removed from her neck. Despite the setback, Ethel Reed arrived in England on 24 September 1927.
It had been a long and distant engagement and although there seems to have been some doubt on both sides, Howard and Ethel married less than a month later on 19 October at Holy Trinity Church in Paddington. The couple settled into life in Cambridge although by all accounts the marriage did not start well. Ethel and Howard had their first child, a daughter, Paquita Mary Joanna, on 26 September 1929. In 1930 Florey was appointed as a very young professor to Sheffield University where the family stayed for five years. A second child, Charles, was born on 11 September 1934. By this time the relationship had deteriorated further as Ethel’s deafness worsened. Both Ethel and Howard, however, were devoted to their children, and despite the growing barrier between them—which they tried to bridge by communicating through notes—they stayed together.
In 1935 Howard Florey was appointed Professor of Pathology at the Sir William Dunn School at Oxford. He was just 37, younger than anyone who had previously held this position. The scene was set. Florey would remain at the Dunn School for the next 30 years providing exceptional leadership and guiding inspired research programs. Being head of the laboratory when he arrived was a daunting task in the economically straitened late Depression years. Florey inherited an impressive laboratory building, a façade really, because he soon found that the Dunn School was not engaged in cutting-edge research, was under-financed, and as a result morale amongst the staff was low.
From the start, Florey had to fight for funds, a task that irritated him because it monopolised his time. In a letter he wrote to the Medical Research Council he expressed his feelings in his blunt Australian style: ‘It seems to me that I have acquired a reputation of being some sort of academic highway robber, because I have to make such frequent applications for grants.’ When he found out that the university planned to cut his grant because of the cost of a new heating plant he wrote, ‘You may gather that I am fed up.’[23]
Despite these economic constraints Florey was determined to turn things around and improve the department’s scientific output. To achieve this Florey adopted an approach that was uncommon in the 1930s. He gathered together a group of researchers who had talent in different fields, an interdisciplinary team where chemists, pathologists, bacteriologists and physiologists could share expertise. Florey realised that science had reached a point where any research project was too big for one person. Gone were the days of Robert Koch beavering away in his own laboratory, carrying out every aspect of his work quietly, secretly and alone.
After Fleming’s discovery of the antibiotic lysozyme Florey and other researchers had turned their attention from chemical drugs to biologically created ones produced by the human body or by moulds and animals. This led to a search in the late 1930s for similar compounds which could kill invading bacteria. After arriving at the Dunn School, Florey made the decision to investigate lysozyme and its properties and began gathering his team. He applied to Cambridge for a biochemist with the appropriate skills to assist him. Many Jewish scientists and intellectuals had been driven out of Germany after th
e rise of Adolf Hitler and the implementation of anti-Semitic policies. Ernst Chain, a gifted biochemist with a difficult personality, was one of them. He joined Florey at Oxford in 1935, appointed as a demonstrator and lecturer in chemical pathology.
Chain had fled Nazi Germany in 1933, some years before the transportation of Jews to concentration camps began, leaving his mother and sister behind in the belief that Hitler was a one-day wonder. He had been pursued by the Gestapo and kept a wanted poster on the wall in his laboratory at the Dunn School.[24] Chain had an incisive mind and from 1935 to 1939 worked on snake venoms, tumour metabolism and the invention and development of methods for biochemical microanalysis. Chain’s innovative approach to a biological problem was to reduce it to its chemical components. It was relationships that were difficult for Chain. His relationship with Florey fluctuated and on occasion he clashed with others on the Oxford team and with university administrators. Because of his background, Chain may also have had to cope with covert discrimination and was perhaps at times treated unfairly.
Working together, Florey and Chain set out to understand the actions of lysozyme. In 1938 Chain solved the biochemical riddle surrounding it. This success led Chain and Florey to the search for other naturally produced antibiotics, a decision which was not regarded as either startling or significant at the time, just a natural extension of their work. It was the confluence of circumstances that led to the penicillin project. Because Florey had somehow wangled some funding from the Rockefeller Foundation in the United States, the two scientists were now sufficiently funded and set out to systematically identify and isolate substances from moulds that could kill bacteria.
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