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Spring 2002

Vol. 02 No. 2


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Comrades in Germs: NDSU scientist seeks Russian help in a quest for a better anthrax test

Obolensk, a clearing in remote woods southwest of Moscow, is the site of a scientific city that has fallen into sad disrepair. The broken windows and grime belie the significance of what once took place there. One building in particular stands out among several drab structures enclosed by an electrified fence, an eight-story box of concrete and glass that houses a collection of hundreds of bacterial strains, many of them genetically altered. Building One, as it's still known, served as the hub of a mammoth Soviet bioweapons program, which at its peak employed 60,000. Within the guarded complex, in one of the most closely kept secrets of the Cold War, Soviet scientists toiled for almost two decades. Their work produced breakthroughs in germ warfare, including deadly neurotoxins, plague and anthrax made resistant to antibiotics.

Inside Building One, on a chilly afternoon last October, Charlie Stoltenow sat at a library table among Russian scientists, some formerly employed in the Soviet Union's biological weapons program, in a scientific exchange. Their meeting was interrupted when a man came in holding news pulled from the Internet, a CNN report that a south Florida man had died from inhaling anthrax, amid spreading fears that he was the victim of a bioterrorism attack. Stoltenow, who knew a little of Obolensk's dark history, asked a blunt question.

"Is this you guys? Are you doing this?"

The Russian scientists looked at each other - an unspoken conversation flickered between them, conveying surprise at the impertinence from the affable, bearded visitor who days earlier had jokingly introduced himself as coming from the North American Siberia.

"Nyet, nyet," came the answer, accompanied by awkward chuckles: No, no.

Stoltenow, an extension veterinarian at North Dakota State University, considered this response for a moment. Maybe he was emboldened by the camaraderie that had developed during the preceding week of meetings, increasingly frank discussions smoothed by small talk over vodka and a trip to the Bolshoi ballet. Or maybe he was thinking of the temptations that might have been dangled before some struggling Russian scientist by someone willing to buy his deadly expertise. He pressed further.

"How do I know this?" Again the Russians exchanged wary looks before one of them, a senior scientist, answered for the group.

"Oh, comrade, if it was us, there would be a lot more dead. This looks like the work of an amateur operation." More nervous laughter followed, this time from the Americans as well, and then the scientists went back to their work, a dialogue exploring research possibilities of mutual benefit. Cooperation in defending against anthrax attacks, in fact, was one of the agenda items in the scientific exchange that Stoltenow took part in, one of two recent delegations of American civilian scientists who met with counterparts in Russia. The respected National Research Council sponsored both trips. Since the collapse of the Soviet Union in 1991, the U.S. government has led programs aimed at keeping the arsenal of biological and nuclear weapons from falling into hostile hands. Even before last fall's attacks, which killed five people, authorities were worried about the lethal use of anthrax, which was available via mail order until the mid-1990s.

ANTHRAX BACTERIA OCCUR NATURALLY THROUGHOUT MUCH OF THE GREAT PLAINS, INCLUDING AREAS OF NORTH DAKOTA AND SOUTH DAKOTA. SOME SCIENTISTS BELIEVE THE BACTERIA MIGRATED WITH CATTLE ON DRIVES FROM TEXAS, WHILE OTHER RESEARCHERS BELIEVE ANTHRAX CAME EARLIER, WITH THE ROAMING BISON.

PULMONARY ANTHRAX WAS FIRST DISCOVERED IN THE EARLY 19TH CENTURY, WHEN WORKERS IN A TEXTILE MILL BREATHED SPORES RELEASED BY NEW MANUFACTURING PROCESSES TO MAKE WOOL. IT WAS KNOWN AS WOOL SORTERS' DISEASE.

Stoltenow, whose specialty is veterinary epidemiology, was selected as part of a six-member team of civilian scientists that included fellow veterinarians as well as several molecular biologists. Previous exchanges involved defense scientists, but the government wants to broaden American-Russian collaborations. During Stoltenow's two-week visit, he was able to see first-hand how even elite Russian scientists must make do with working conditions one would expect to find in the Third World. Dilapidated buildings at Obolensk were dotted with broken glass panes, some replaced by boards or aluminum foil. For the first 10 days, Stoltenow's hotel room had no heat. Obolensk had just two large apartment buildings, no business district and few services. Within the science compound, laboratories function with outdated equipment. On a tour of one lab, Stoltenow's group saw an empty gin bottle under a lab-bench ventilation hood. Most surprising, he says, was the fact that it was left out in the open, where it could be seen.

Earlier exchanges, involving U.S. military or other government scientists, often ended in mutual frustration. Sometimes scientists agreed to share information, only to be overruled by their superiors. Russian scientists, for instance, once agreed to provide copies of their arsenal of germs - specimens from Building One's Museum of Cultures, a deadly collection of plague, anthrax and tularemia microorganisms - but security officials nixed the idea. Almost five decades of Cold War hostility will not evaporate overnight, in spite of a warming in relations. The scientists work under watchful eyes. Case in point: Stoltenow learned that one of the Russian scientists, a balding, bearded man whose vague explanations of his duties seemed to change daily, was, in fact, a spy for the Russian equivalent of the FBI. The suspicion is mutual. While in Moscow, en route back to Fargo, Stoltenow encountered some American military officers in an international hotel and joined them for dinner. He told them about his talks with Russian counterparts, with hopes of cooperating to combat bioterrorism. The officers shrugged, saying such lofty work is best left to civilians. "In two or three years we could be at each other's throats - you never know how world politics will go," he says, quoting the officers' reaction.

The changes have, indeed, been staggering. During the late 1980s, at the peak of the Soviet bioweapons program, Obolensk teemed with a staff of 3,000 scientists and technicians. The number today has dwindled to approximately 1,000. Among those who remain, even top scientists are paid the equivalent of $500 a month. Stoltenow knows of one former scientist who works as a construction worker; another served as one of their interpreters. In light of the widespread poverty and lack of jobs, Western officials worry that some unemployed Russian scientists might work for terrorists or rogue states. "I asked them 'where are the other 2,000,' " Stoltenow says of the scientists who left Obolensk. "They said 'we don't know.' It was pretty scary. People are a pretty liquid commodity. What type of information left - who knows?"

As demonstrated by last fall's bioterrorism-by-mail attacks, the first evidence of a clandestine anthrax release is likely to be stricken patients turning up in emergency rooms, complaining of symptoms similar to pneumonia or other respiratory infections. Unfortunately, by the time symptoms turn up in cases of inhaled anthrax, it is often too late for a cure. The fatality rate of pulmonary anthrax approaches 95 percent if treatment doesn't begin within 48 hours of exposure. Blood culture tests are used to diagnose the disease, usually evident within 6 to 24 hours. Confirmatory immunological and microbiological tests can take up to several days to provide a definitive diagnosis. That leaves little margin for error.

However, Stoltenow says, the tests used for initial diagnosis of anthrax are prone to false positive results, causing unnecessary panic and expense. Given those problems, he has assembled a team of scientists, Russian and American, to search for a new testing method for anthrax. The technique he proposes would zero in on detecting what is called the lethal factor of anthrax, a toxic protein that must be present for the bacteria to cause a fatal exposure. Once inside a nutrient-rich environment - such as warm, moist lungs - the bacteria secrete the lethal factor and two other proteins, a protective antigen and an edema factor, which causes a rapid buildup of fluids in the victim. Luckily, researchers have learned that anthrax with the lethal factor develops a distinctive cleavage of protein enzymes once it establishes itself in a host; by targeting that long cleft, using biochemical processes that "see" unique molecular shapes, Stoltenow and his colleagues hope to develop a more reliable medical test to confirm the presence of anthrax.

Three prospective Russian colleagues from Obolensk have expertise in virulent anthrax strains and their proteins. His four American collaborators would include Lynn Rust, a microbiologist at NDSU; Eric Garber, a biochemist with the U.S. Department of Agriculture in Fargo; as well as a diagnostic molecular biologist with expertise in developing test kits from South Dakota State University and a Colorado veterinarian whose company is the sole supplier of anthrax vaccines for livestock.

AN ANTHRAX INFECTION REQUIRES EXPOSURE TO BETWEEN 10,000 AND 20,000 SPORES, A MICROSCOPIC QUANTITY. AS SOON AS AN ANTHRAX SPORE ENTERS THE BODY IT GERMINATES AND MULTIPLIES. WITHIN A FEW DAYS, THE BACTERIA PRODUCE TOXINS THAT CRIPPLE THE ABILITY OF WHITE BLOOD CELLS TO FIGHT DISEASE.

THE PIN CUSHION PREVENTION: ANTHRAX VACCINE SHOTS MUST BE GIVEN SIX TIMES BEFORE THEY BECOME EFFECTIVE - THREE TIMES IN TWO-WEEK INTERVALS AND THREE TIMES IN SIX-MONTH INTERVALS, FOLLOWED BY YEARLY BOOSTERS. SO FAR, THE VACCINE IS NOT AVAILABLE TO THE GENERAL PUBLIC.

"Quite frankly I wanted to do something out of the Midwest," Stoltenow says. "We're small schools, but we have excellent scientists." His proposal, for funding to bring the Russian and American collaborators together for an initial strategy session in Washington, D.C., is pending before the Department of Defense. In addition, Stoltenow is formulating a second research proposal, involving anthrax monitoring and surveillance, to collaborate with a colleague who is an anthrax expert working for the Russian equivalent of the Centers for Disease Control. The plan is to study environments where anthrax occurs naturally - as it does in three pockets inside North Dakota as well as nearby northwest Minnesota - in the hope that someday it will be possible to predict where outbreaks might occur.

Stoltenow is pleased with the rapport he has developed with his Russian colleagues. He has bilingual business cards, with English on one side and Russian on the other, and keeps in regular contact with Russian colleagues via e-mail. "I'm very encouraged about the Russians," he says. "They were very matter of fact, yet very open," and many have sons and daughters studying in American universities. "We don't have to fear the Russians." Still, he would like to learn more of the secrets kept inside Building One, which he fears is a microbiological Pandora's box. "It would really be good if we could know what's in their arsenal," he says. "If we know what's in there we could have a pretty good idea of what might have slipped out of there."

- Patrick Springer


Student Focused. Land Grant. Research University.