Two Scientists Tracing Anthrax Family Tree Investigate Origins of Strain From Attacks

By ANTONIO REGALADO, Staff Reporter of THE WALL STREET JOURNAL

When Kimothy L. Smith heard CNN report that a man in Florida had been diagnosed with the inhaled form of anthrax, he raced to his laboratory at Northern Arizona University and "cleared out the incubator," he says. After making way for incoming samples, he then waited for the call for help from federal officials.

Locked in an unassuming two-story campus building in Flagstaff, the Arizona laboratories house the world's largest collection of specimens of anthrax bacteria, about 1,350 in all. For several years, the lab has been undertaking one of the most ambitious efforts to catalog anthrax DNA and trace the origins of different strains.

Because of that work, Dr. Smith and his boss, Paul Keim, have been crucial players in the most intensive bioterrorism criminal investigation ever in the U.S. It was here that anthrax samples recovered from American Media Inc., NBC News, Sen. Tom Daschle's offices and elsewhere were sent to be genetically dissected. Drs. Smith and Keim won't comment on their work in the investigation, but sources close to the research say the Arizona team determined that the anthrax bacteria from these different sites are indistinguishable and have "Made in the USA" stamped all over them: The microbe is a descendant of some taken from a sick cow decades ago near Ames, Iowa, and thus dubbed the Ames strain.

Investigators hope that identifying the strain will help pare the list of possible perpetrators. The Ames strain has been studied extensively by the U.S. military during the past two decades and has never been traced to Iraq or the former Soviet Union, lending support to theories that a homegrown bioterrorist is at work.

Investigators say they still don't know where the anthrax came from, or whether the bioterrorists are foreign or domestic. They don't rule out the possibility that foreign bioterrorists or states got hold of the Ames strain, perhaps from a U.S. source. And complicating the scientists' efforts to sort out the trail of anthrax ownership is the history of anthrax research itself, confused by incomplete records and institutional secrecy.

The 46-year-old Dr. Keim started the campus laboratory at his alma mater in 1989 and began his work on anthrax genetics while on leave at Los Alamos National Laboratory. Earlier this year, his lab proved that the Japanese doomsday cult Aum Shinrikyo had failed to cause any casualties in a protracted 1993 anthrax attack because they had used a harmless strain used in vaccines.

Dr. Smith, a former practicing veterinarian in Altus, Okla., decided to move into research after the routine of tending to cows, dogs and the occasional raccoon wore thin. At Louisiana State University's Special Pathogens Laboratory, he found his niche in molecular epidemiology, the hunt for the cause of animal-disease outbreaks among the proteins, enzymes and DNA that pathogens such as anthrax use to survive in the wild and infect their hosts. In 1999, he moved to the Arizona lab, bringing with him the lab's first samples of anthrax.

Since then, the Arizona lab has been building a genetic family tree of all forms of anthrax. Under a $300,000-a-year grant from the National Institutes of Health, Drs. Keim and Smith have been collecting anthrax varieties from across North America and Africa and then examining them for patterns of unique DNA markers.

The job of collecting field specimens falls largely to Dr. Smith, a 40-year-old with a long mane of silver hair who steals quick puffs from a pipe he keeps tucked in his jeans. He has made several trips to South Africa to pick anthrax specimens from animal bones, hyena dung and occasionally from local farmers who caught the skin form of the disease while preparing hides. Among his most prized acquisitions: two brown glass tubes of anthrax sealed in 1937 by Max Sterne, the South African who developed the first effective anthrax vaccine.

Telling varieties of anthrax apart isn't easy. Compared with other microbes, anthrax displays scant variety at the DNA level. Part of the reason is that anthrax takes regular breaks from evolution. After killing their host, anthrax bacteria form protective spores and enter a state of suspended animation that has been known to endure more than 150 years. That's what makes anthrax a favorite of weapons makers and terrorists: It can be stored indefinitely, in a warhead, for instance, or an envelope.

Now, as scientists and investigators try to trace the anthrax that has appeared since Sept. 11, they are running up against the shortcomings in anthrax research and record-keeping before DNA typing was developed. Until recently, microbiologists kept track of their collections with handwritten labels, and they relied on colleagues to maintain the paper trail when they shared specimens.

The original Ames strain sample came from a dead cow, but no one knows exactly when. Martin Hugh-Jones, a scientist at Louisiana State University, Baton Rouge, says the sample his lab obtained in the mid-1990s was marked "October, 1932." That sample came from the Center for Applied Microbiology and Research in Porton Down, England. Porton Down got its sample from the U.S. Army Medical Research Institute of Infectious Diseases, which got its own sample in late 1980 or early 1981 from the U.S. Department of Agriculture's veterinary laboratory in Ames, Iowa. The USDA doesn't know how long it has held the sample.

It also isn't clear whether the Ames strain was part of the secret U.S. biological-weapons program, to which President Nixon called a halt in 1969. Dr. Hugh-Jones has said that David Huxsoll, former head of the U.S. Army Medical Research Institute of Infectious Diseases, told them it was. Dr. Huxsoll couldn't be reached for comment. Two scientists who were involved in the U.S. bioweapons program, Bill Patrick and Richard Spertzel, say they used a strain called Vollum, not Ames.

What is certain is that during the past 15 years, the Ames strain, particularly virulent, has become a research favorite. The Centers for Disease Control and Prevention has refused to release its records of other U.S. labs that may have Ames, citing national-security concerns. Toward the middle of October, the Federal Bureau of Investigation subpoenaed all the records of Dr. Hugh-Jones, asking for the names of everyone who had visited his laboratory.

Soon after the first press reports that the anthrax used in the recent attacks was of the Ames strain, Iowa State University briefly became the prime suspect in a media hunt for the source. Gov. Thomas Vilsack called out the National Guard and state police to guard the anthrax stores. Iowa State researchers got rid of the problem by baking their anthrax specimens in an autoclave, a type of oven used to sterilize surgical instruments.

Iowa State says it received permission from the FBI and the CDC to destroy the anthrax. But now, no one will ever know whether the Iowa State anthrax was the Ames strain -- perhaps even the original. That dismays Dr. Smith. The dusty collection would have been a precious data point for his anthrax catalog. "It sounds like a damn convenient thing to autoclave it," he says.

There are indications that the terror strain differs in some small but significant way from the Ames strain held by the U.S. military. Tom Ridge, President Bush's director of homeland security, has called the strains used in the Florida, New York and Washington attacks "indistinguishable" but not identical. Anthony Fauci, head of the National Institute of Allergy and Infectious Diseases in Bethesda, Md., said on Saturday that the strain was "Ames-like."

Such differences may matter in the investigation. Each time a bacterium divides, it has to make a new copy of its DNA, and there's a small chance for error in the process. In experiments two years ago, Dr. Keim and his colleagues found that Ames is exceptionally stable. After growing a sample until it had doubled 100,000 times, they found only one detectable genetic change. If the Arizona scientists and others find that the terrorist anthrax exhibits such differences, that could indicate that it has been grown in huge quantities, potentially a sign of involvement by a state-sponsored biological-weapons program.

Because Ames originated in the U.S. and probably persists somewhere in the natural environment, Dr. Keim doesn't discount the possibility that a lone American terrorist in a Western state might have lifted anthrax from a dead deer or bison.

Dr. Smith agrees that right now, most explanations are possible: "The word on the street is that nobody has a clue where this comes from."