Experiment Eleven

by Peter Pringle

  Woodruff had found the college experience exciting and sometimes a little overwhelming. He had gone to a concert for the first time, and had celebrated with his fellow students all night in 1937 when the Rutgers football team scored its first victory, 29 to 27, over Princeton since 1889. He graduated in soil chemistry, and Waksman offered him a college fellowship of $900 a year—20 percent more than fellowships elsewhere. The money came from Merck’s generous contributions to Waksman, now totaling $3,600 a year.

  As a result of his European tours, Waksman attracted students worldwide. Eleven graduate students crammed into the two upstairs laboratories of the Department of Soil Microbiology. They came from China, South America, Europe, and across the United States. At first Woodruff was “terribly discouraged” when Waksman put him to work on composts and gave him little direction. He relied heavily on Waksman’s deputy, Robert Starkey, as had most of Waksman’s graduate students over the years. As one of them recalled, Starkey was their “great provider of materials and receiver of complaints—the equivalent of an assistant in a steel mill. He remembered; he got things done. He told us how to make our cases to Dr. Waksman. Modest to a fault, totally loyal to the Department—it’s inconceivable that Dr. Waksman could have operated without him.” Woodruff worked alongside a visiting student from China who was trying to find out the minimum temperature needed to kill all the harmful bacteria in human feces so that it could be used for compost. Since human feces were not used for compost in America, Waksman suggested that Woodruff should study horse dung, horse urine, and straw, and see how the combination worked. After that, Woodruff moved on to potato scab disease, a serious problem in New Jersey, caused by an actinomycete. The microbe does not grow under acid conditions and is controlled by farmers by adding sulfur to the soils. A bacterium oxidizes the sulfur to sulfuric acid, thus producing the desired soil acidity.

  One day, toward the end of 1939, Waksman received word from Merck that the Oxford team was successful in isolating and purifying penicillin. Highly agitated, Waksman appeared in the laboratory and told Woodruff to “drop everything. See what these Englishmen have discovered a mold can do. I know the actinomycetes can do better.” Certainly, the Russians had already suggested that the actinomycetes were worth testing.

  Waksman took Woodruff to see René Dubos at the Rockefeller Institute so that he could learn the so-called soil-enrichment method that Dubos had used when he discovered tyrothricin. In this method, the researcher adds a disease-producing bacterium to pots of soil over two to three months, hoping this will favor growth of the species of actinomycetes that can kill and feed on that particular bacterium. Woodruff chose E. coli as his disease bacterium, adding billions of cells to pots of Rutgers college farm soil he knew was rich in actinomycetes. Each week he counted the surviving E. coli by taking a sample of the soil in the pots and growing it on nutrient jellied agar in a petri dish. It was easy to spot the E. coli, which appeared as clumps of mold with a distinctive metallic luster. Each time he counted them, the number was reduced until at the end of the three months Woodruff’s pots of enriched soil had, as he put it, “become highly efficient E. coli killing machines.” He isolated the sturdy-looking cultures of Actinomyces antibioticus, which produced a red chemical substance apparently responsible for the killings. He had found a new antibiotic. Woodruff and Waksman named it actinomycin.

  Antibiotics are often difficult to extract and purify, as Fleming had found with penicillin, but actinomycin was relatively easy. Woodruff and Waksman hoped they had struck lucky, but now they needed to make enough actinomycin to study its effect on disease in small animals.

  Waksman took a sample to Merck, where the new drug could be produced on a larger scale than in his laboratory. Then he and Woodruff wrote up their results in a paper that was published in the spring of 1941. Merck’s researchers purified and tested actinomycin in animals. The results were shocking. It certainly killed disease bacteria, but it was so toxic it also killed laboratory mice in twenty-four to forty-eight hours. There was no question of testing it on humans. Rat poison was about all it was good for.

  Waksman was “truly excited” about the discovery, however. “Once I had an actinomycete in my hand which produced antibiotic activity, everything changed,” Woodruff recalled. “Waksman started coming into the lab immediately after lunch each day.”

  At this early stage, these microbial chemical weapons were so new they had no specific name. They were known as “antibiotic substances.” The term came from “antibiosis,” meaning “against life,” a word first used by Louis Pasteur’s pupil Jean Paul Vuillemin in 1889 to describe the antagonistic effects of microorganisms. By the 1930s, the use of “antibiotic” as an adjective, as in “antibiotic substance,” was fairly common in the scientific literature, especially among European researchers. Waksman himself used it. But now a proper noun was needed.

  At one of Waksman’s Friday afternoon meetings when students discussed the latest scientific journal articles, Waksman asked the gathering for a noun to describe the new wonder drugs. The students offered some suggestions, but Waksman already had an answer: “antibiotic.” He even had a definition ready. An antibiotic, he said, was “a chemical substance produced by microorganisms, which has the capacity to inhibit the growth of and even destroy bacteria and other organisms.” The noun entered the medical lexicon, but Waksman’s colleagues would argue furiously whether he should be credited with “coining” the word, or simply applying it as a noun. The debate continues.

  Now armed with the redefined word, Waksman put his students to work, screening soils and composts for the new candidate antibiotics. One of these researchers, a postdoctoral student named Walter Kocholaty, arrived at Rutgers from the University of Pennsylvania to learn the methods Waksman was using. Instead of soil enrichment, which took up to three months to produce a likely microbe, Waksman now switched to a second procedure to save time. Tiny amounts of farmyard soil, or compost, were diluted in tap water and drops of the soil and water mix put into a petri dish containing living cells of a disease bacterium in jellied agar. The disease bacterium was the only source of food for the microbes from the soil and water mix. As these microbes grew, the ones capable of killing and eating the disease bacterium created the tell-tale clear zones in the agar. Kocholaty found a strain of a purple-colored actinomycete, A. lavendulae, produced good clear zones in the petri dish.

  Waksman had first come across A. lavendulae in his early research in 1915 and Kocholaty now isolated robust-looking strains from the colonies growing in the petri dish, transferred them to separate dishes, let them grow, and then tested the strain against more disease-producing bacteria by the “cross-streak” method. This involved “streaking” a line of the A. lavendulae strain down the right-hand side of a petri dish and then streaking the bacteria to be tested at right angles to it—like bringing troops up the front line. When clear zones appeared in the lines of disease bacteria, Kocholaty knew he had a new antibiotic in A. lavendulae.

  His time at the lab was over, however, and Waksman gave the strain to Woodruff for further tests. He grew the strain of A. lavendulae in various different broths to see which one would case the strain to produce the most antibiotic. Waksman and Woodruff named the antibiotic streptothricin, after Streptothrix, or “twisted fungus,” the name given to an actinomycete by a nineteenth-century German researcher. Woodruff made enough streptothricin for animal tests in the only lab available at the Rutgers college farm—the dairy. There, they found it seemed to cure cows of a bacterial disease, brucellosis, that caused contagious abortion. Excited, Waksman and Woodruff now took their second antibiotic to Merck, where company researchers confirmed their findings. Streptothricin looked like a winner. For the last six months of Woodruff’s Ph.D., Waksman sent him to work on streptothricin at Merck, where he could be Waksman’s “eyes and ears” as the company continued its testing.

  Merck was so excited by streptothricin, the company even built a small production plant. It cured bacterial
infections in mice, but when company researchers started a clinical trial on four human patients, they found that almost immediately the patients’ kidneys stopped producing urine, so the treatment was stopped. “We thought they were all going to die,” Woodruff recalled. “We were all worried for a couple of days.” Woodruff was devastated. The failure of streptothricin was the greatest disappointment of his life, he would say later. But he was fortunate; Merck offered him a job and he stayed at the company until retirement.

  Despite the two failures of actinomycin and streptothricin, Waksman was determined to keep his search going. The ranks of his students available to do the screening were dwindling, however. The draft board was now taking staff from him, mostly men. When Woodruff arrived at the department there were eleven graduate students; by the middle of 1942, there were only four. One of them was a skinny youth, a recent Rutgers graduate named Albert Schatz.

  By the time Schatz arrived in the early summer, the graduate students had isolated some 400 cultures of actinomycetes and 160 of fungi and were concentrating on four microbes that produced antibiotics: two from actinomycetes, actinomycin and streptothricin, and two from fungi, clavacin and fumigacin.

  Schatz had wanted to do his Ph.D. on soil science but the department head was not as good a salesman as Waksman, didn’t have a commercial product, and could not attract any funding. Schatz lived for free in a room off the Plant Pathology greenhouse in return for minor duties tending the experimental plants grown to fill President Roosevelt’s Victory Gardens. Like Boyd Woodruff, he lived off eggs from the poultry shed and milk from the dairy.

  Waksman’s reduced team of four graduates was happy to work on antibiotics under its “fatherly professor,” a short man, by now slightly overweight, with a bristly mustache and bright, intelligent eyes. His clothes were never pressed, and his vest always carried some evidence of a recent meal. The young students, like Schatz, admired Dr. Waksman, even idolized him, referring to him—only behind his back, of course—as “Old Waksie,” “the Boss,” and sometimes “the Great White Father.” “The atmosphere was terribly stimulating,” recalled Doris Jones. A male former student recalled, “One of his [Waksman’s] favorite tricks was to stop you in the hall and ask how everything was coming along. As you started to explain something, he would say, ‘Good, let us not waste any time, follow me.’ He would then take you into the men’s room where you continued your scientific report while he was not wasting any time.” His European colleagues regarded him as “a typical representative of the Russian Jewish intellectual at its best.” Schatz started work immediately on experiments on streptothricin and another antibiotic, clavacin, that had been found to be toxic in animal tests.

  The war was taking its toll on Waksman’s team. By July, all except Schatz had left to go into industry or teaching, which offered a draft deferment—Boyd Woodruff had gone to work at Merck, for example—or had been drafted into special branches of the military where their lab experience could be useful. Waksman’s antibiotic project was struggling for lack of funds. Seeing that the federal government was funding the production of Fleming’s penicillin, and now Dubos’s gramicidin, he applied to the government’s Committee on Medical Research, which had been set up to fund wartime medical projects. But he was turned down. The committee was looking for projects that could produce results “within relatively short periods of time.” Penicillin and gramicidin had already proved their worth. Waksman’s project covered “a broader field” that was, at that moment, more theoretical than practical. The committee suggested that Waksman try philanthropic foundations, and it passed his application to the Commonwealth Fund, in New York. He was soon awarded a grant of ninety-six hundred dollars—more than enough to add equipment and research staff, if he could find researchers exempted from war service.

  In October 1942, after five months of working for Dr. Waksman, Schatz received his draft papers. Waksman tried unsuccessfully to persuade the draft board that Schatz was “more valuable” working on the search for new antibiotics than “becoming a private in the army.” He then pleaded with the draft board to at least give Schatz a job related to his studies, and in November 1942, Schatz became a laboratory technician at an Army Air Corps hospital in Miami Beach. There, thousands of miles from the front lines, he was exposed to the terrifying effects of war. He drew blood from seriously wounded and diseased soldiers brought home to America, and he identified the microbes responsible for their infections.

  It was relatively simple work, easily accomplished, as he was already something of an expert. But Schatz quickly learned that once he had identified the germs, the army often had no medicines to cure the diseases. Penicillin was being produced as a war priority but was still in short supply, and its healing powers were strictly limited. It had no effect on the germs that cause illnesses epidemic in wartime, such as cholera, typhoid, urinary and intestinal infections, and, the persistent killer, tuberculosis. Schatz knew which soldiers in the military hospital had no hope of recovery, and in his off-duty hours he volunteered to sit by their bedside, comforting them as best he could in their last hours.

  As news from the front improved in the spring and early summer of 1943 and the Allies were preparing to invade Italy, Schatz was hopeful of an early return to his studies. He was homesick for the camaraderie of Dr. Waksman’s close-knit group. He wrote to Waksman as frequently as his duties permitted and complained about the monotony of military life, saying that he hoped to return to his studies soon.

  “All of us are eager for the war to end,” he wrote in one such letter. “I personally can hardly wait to get back home and return to school. Regards to all, Very respectfully yours, Pvt. Albert Schatz. P.S. I hope all of Africa is in Allied hands by the time this letter arrives.” Waksman, always encouraging, suggested he should look for antibiotics in the Florida soils, and Schatz mailed samples of promising candidates to Waksman for his microbe collection.

  His return to Rutgers came even sooner than he had hoped. As he was loading supplies onto a truck, he twisted his back, and army doctors discovered he had more than a sprain. Private Schatz had a malformation of the spinal cord, apparently undetected during his draft board medical exam. He was declared unfit for military service and honorably discharged. The army recommended him for a Good Conduct Medal, which he never actually received, and gave him the train fare home to New Jersey on June 15, 1943.

  Pvt. Albert Schatz, U.S. Army Air Corps, Medical Detachment, in Miami Beach, Florida, in 1943. (Courtesy Vivian Schatz)

  Waksman quickly reenrolled him in the antibiotics program, but he could only pay forty dollars a month, a third of what Schatz had received before he went into the army. Schatz didn’t mind. Despite his back injury he was, if anything, more self-assured, more independent, and he returned to his free room off the Plant Pathology greenhouse. He was ready to start working round the clock, sleeping “in his cloth,” as he put it.

  5 • A Distinguished Visitor

  IN SCHATZ’S EIGHT-MONTH Absence, Waksman’s team, which now included two new women graduates—Doris Jones and Betty Bugie—had not found a single new antibiotic. But they had had lots of fun trying, according to Jones. She loved “the close-knit group, the Friday brown-bag lunches where the smell of sandwiches mixed with the scents of the molds, especially the earthy odor of the actinomycetes.” A bright, jolly, optimistic person with a self-deprecating wit—she once described herself as having “well-padded bones” and acknowledged her nickname, “Moose,” because of her loud voice—she didn’t mind the hard work. “We call the laboratories the Salt Mines because in order to pull a practical antibiotic producer out of Mother Nature we literally have to ‘work our asses off,’” Jones recalled later. But she appreciated how Dr. Waksman urged his graduate students not “to waste time studying extraneous things in books. I worship him.”

  Merck researchers were testing the most promising microbe so far—streptothricin—but it was becoming apparent to Waksman that streptothricin’s toxicity could not be su
fficiently reduced, even in the purer form made by the Merck chemists, to “offer hope for its eventual usefulness.” So when Schatz resumed working on his Ph.D., the opportunity to make his mark on medical history was wide open. And Waksman, for the first time, was interested in testing his microbes against the genus Mycobacterium, some species of which can cause tuberculosis.

  A year earlier, he had been pressed to start such a project by his son, Byron, who was then finishing his degree in bacteriology at the University of Pennsylvania Medical School. In a letter, Byron had offered to do a summer project at Rutgers, first testing microbes against the nonvirulent strains of Mycobacteria in Waksman’s lab and then, if an active agent was found, testing it against the virulent human form in animals. As Waksman did not have animal-testing facilities in his department, that risky second test would have to be done elsewhere. But Waksman was still not ready. “The time has not come yet,” he replied to his son.

  In their competing versions of what happened next, both Waksman and Schatz later claimed that they were the one to first take the TB project seriously. Waksman said he finally decided to go ahead with the project “early in 1943.” Schatz said that on his return from the army in June he knew exactly what he wanted to do for his Ph.D., and he chose the most ambitious research project that any Waksman graduate had ever suggested. He would find a new antibiotic that would cure the diseases his military comrades had died from, and that included tuberculosis.

  The evidence does not resolve this disagreement, but seems to favor Waksman. He was certainly moving closer to testing his antibiotics against the TB germ. On June 1, according to his expense records, he went to New York City to meet Dr. Leroy Gardner of the Trudeau Sanatorium at Saranac Lake. The topic was “the problem of bacteriostatic substances in relation to tuberculosis.” On June 18, Waksman wrote to Dr. Florence Seibert at the Henry Phipps Institute in Philadelphia, where they carried out research on TB. Dr. Seibert had invented the first reliable tuberculosis test for humans. Waksman asked for fifty to one hundred grams of dried cells of the human TB H37 germ, and also a culture which could be used for growing the organism. Dr. Seibert sent one of each, but warned that the culture was old and of uncertain viability—the dried cells had been alive nine years before being dried in 1943. In other words, she did not know whether these strains would produce meaningful results if used as a test for TB.