The tea-time question “One lump or two?“ is innocuous. If, however, the sugar cubes in question were in the possession of Baron Otto Karl von Rosen, a prudent response to the familiar query would be “none.“

The baron was a spy and saboteur for Germany during World War I. His sugar cubes were altered to produce a lethal aftertaste. They were loaded with anthrax.

Remarkably, some of the baron's bacilli have been revived after eight decades of dormancy. The microbial resuscitation represents a closing chapter to one of the few well-documented instances of a deliberate plan to use bacteria as weapons of war.

“I think they are the oldest known spores to be revived with any certainty,“ Richard Manchee, senior microbiology advisor at the Defense Evaluation Research Agency at Porton Down, in Salisbury, U.K., told BioWorld International.

Anthrax spores have been revived from 200-year-old bones found in Africa. “But,“ Manchee noted, “anthrax is an endemic disease in Africa. They [the spores] could have got there subsequently. They occur commonly in the soil.“

When animal or human victims are buried, anthrax leaks into the soil and survives harsh conditions as a spore. (See BioWorld Today, May 1, 1998, p. 1.)

Humans, surprisingly, were never the intended victims of the baron's biological warfare. The targets were reindeer and horses. The animals were used to haul British supplies across northern Norway and Finland to Russia during the war.

The Germans may have approved the use of anthrax to destroy the animals and hinder the flow of supplies to their enemy. The baron evidently was working for the Germans when he and some companions traveled to northern Norway in 1917 equipped with dynamite, curare, unidentified microbial cultures and 19 anthrax-laden sugar cubes. Each sugar cube contained a small hole into which a glass vial of anthrax culture had been inserted.

A pair of those sugar cubes recently turned up in a vault in a police museum in Trodheim, Norway. One cube still held a tiny glass vial containing a few microliters of brown fluid.

“I'm assuming the liquid was [originally] a heavy suspension of anthrax spores, the original culture probably,“ Manchee said.

After reading a brief description attached to the exhibit, the museum curator passed the disturbing curio - which appears to be the only remaining anthrax-laden sugar cube from the baron's stock - to Norwegian authorities. They contacted Manchee.

In a scientific correspondence titled “Deadly relic of the Great War,“ in the June 25 issue of Nature, Manchee and his co-authors describe how they revived the dormant pathogen using enrichment, a technique routinely used to revive fading bacterial cultures.

Although the number of surviving bacteria was very small and the culture was on “the very edge of viability,“ the work underscores the remarkable hardiness of anthrax spores and the potential problems posed by long-term contamination following use of this deadly agent of biological warfare. Under better conditions, anthrax spores might be able to survive even longer than 80 years.

“These certainly were not stored under ideal conditions: stuck away in a drawer in a museum. Had they been refrigerated they probably would have survived in a better state,“ Manchee said.

Also known as woolgatherer's disease, anthrax commonly affects livestock. In humans, it has two distinct phases. The first is characterized by flu-like symptoms and congestion in the chest. If detected early, it usually can be treated with antibiotics.

A second, terminal phase follows several relatively uneventful days. This deceptively quiet time ends as large amounts of toxin are released into the body by the growing anthrax bacilli.

The ability to form tough spores distinguishes Bacillus anthracis from many other candidate bioweapons. This, in combination with its lethality, has made it a favorite among countries developing biological weapons. The spores can be delivered in an aerosol spray.

The first scientific evaluation of B. anthracis as a potential bioweapon took place in 1942 on Gruinard Island off the coast of Scotland. Using sheep as subjects, the British military demonstrated that anthrax spores could both survive the explosion of a bomb used to disperse them and remain lethal.

In the November 19, 1981, issue of Nature, Manchee and colleagues described the results of a 1979 survey of still-viable anthrax spores on the island. In the November 1994 issue of Applied and Environmental Microbiology, he and his co-authors described the successful inactivation of the spores on Gruinard using formaldehyde. The spores had survived on Gruinard for more than four decades.

“Most candidate bioweapons are pretty unstable, really - especially in storage - but, as we all know, anthrax spores can persist for many years,“ Manchee said.

Baron von Rosen was arrested by Norwegians before he was able to complete his mission. Plans to use anthrax as a weapon, however, still continue. The U.S. government considers anthrax such a threat from renegade countries it is vaccinating all service personnel against the disease.

Vaccines produced in the U.S. and the U.K. use one component of the anthrax toxin called “protective antigen.“ These vaccines sometimes produce mild side effects such as malaise and soreness around the inoculation site. *

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