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Anne Camper said the military is interested "big time" in sensors to detect E. coli in water. 		Biohazard

Water and the "bad" E. coli

The idea of spreading disease through water is as old as the first tribesman to put a dead animal into a creek upstream from a rival clan. Today there's a reason not just any one is allowed access to municipal water systems that serve hundreds, thousands, even millions of people from a central source.

Having said that, MSU microbiology professor and associate dean of graduate studies Anne Camper is quick to point out two barriers to spreading a bioterrorist agent through water systems, assuming a terrorist could get inside a guarded water treatment plant.

The first is quantity. Because water disperses and dilutes, a person would need a lot of a potential agent to create illness (maybe a truckload, Camper thought). Second, municipal water systems carry disinfectants that are going to have some effect on potential contaminants.

"This isn't something you could do on a whim," Camper said. "The risk is low, but it's there."

Here's a scenario that Camper and the American Water Works Association Research Foundation (AWWARF) thought warranted some looking into about six years ago:

What would happen if a pathogen like the bad E. coli-known as the hamburger E. coli for the deaths it caused several years ago at a Jack-in-the-Box restaurant-got into a water system and "interacted" with slime called biofilm?

To put it a different way, what if colonies of harmless bacteria (called biofilms) that often dwell in water systems, like the bacteria that harmlessly inhabit the human gut, were to trap pathogens and shield them from disinfectants? Could the biofilms become reservoirs for disease?

The question isn't a theoretical one. Last year an Ontario city had the bad E. coli (E. coli 0157:H7) in its municipal water system. Run-off from cattle waste was the suspected source. Similar instances have occurred in New York and North Dakota.

Camper's studies were somewhat inconclusive. It appears that E. coli 0157:H7 might accumulate in a biofilm, but it won't grow there. Over time the numbers would disappear, but just how quickly depends on the water system.

Building on that research, Camper and MSU colleague Paul Stoodley began investigating how to improve the ability of sensors to detect "transient events," or a few dangerous microbes like E. coli 0157:H7 flowing through a huge volume of water.

Working with the Butte environmental engineering company MSE, the scientists' goal is to increase the sensitivity of existing sensors without creating a lot of false positives. The idea is to incorporate a biofilm-based technology-something that takes advantage of the harmless bacterial clumps often found in water systems- into current sensors.

The military is interested "big time" in what the Montana scientists are trying to develop, said Camper, which is why the MSU Center for Biofilm Engineering has applied for additional funds from the defense organization called DARPA.

Camper predicted a new biofilm-based sensor could be within reach within a few years and would be unique. She knows of no other research groups coupling biofilm-based technology with existing water sensors. Indeed, MSU's biofilm center is one of just a handful of such centers worldwide.
 

Michelle Hardy is learning about the non-deadly but disruptive Norwalk virus. The military is interested in a vaccine.
Biohazard

Norwalk

A Navy seaman goes on leave. Without knowing it, he picks up a gastrointestinal illness caused by the Norwalk virus, and by the time he has symptoms-vomiting, diarrhea-he's back on the aircraft carrier. Although not deadly, Norwalk virus is highly communicable through improper hygiene (not washing hands after using the bathroom). Before long several hundred crew members are down for the count.

Sound like the plot of a C-rate war novel? Something like this really happened during the Gulf War, said MSU assistant professor of veterinary molecular biology Michele Hardy.

"Outbreaks are common on aircraft carriers and other closed military settings because it's so easily spread from person to person," Hardy said.

Norwalk virus is also spread through contaminated food and water. A 1999 study suggested that the virus may cause more outbreaks of food-borne illness than bacteria and parasites. One sick bakery worker, for example, was linked to an estimated 3,000 cases of gastroenteritis in Minnesota one year, according to the Centers for Disease Control.

Symptoms usually pass in three or four days but a person can infect others for up to a week after feeling better. That's why nursing homes, restaurants, summer camps, day care centers, hospitals, sports teams and families all provide the perfect environment for the virus to spread.

Although the virus is not a potential bioterrorist agent like anthrax or brucella, Hardy's learning all she can about the virus with the hopes of pointing researchers toward a vaccine. She said the military is especially keen on the idea of vaccinating everyone before deployment. Another goal is to develop medication for those already sick and for those exposed to the sick person.

The virus, however, is not going to give up its scientific secrets easily. Discovered in 1972, Norwalk virus wasn't cloned for almost another two decades. Cloning it for study was a major milestone, as the virus won't grow in the lab or in animals as other microbes will. It only grows in people.

Hardy is working with LigoCyte Pharmaceuticals, Inc., in Bozeman on the Norwalk vaccine. LigoCyte research focuses on the molecules that allow disease-causing organisms like Norwalk virus to stick to cells inside the body. It's this sticking-called bioadhesion-that starts the disease process. Hardy's role is to identify those bioadhesion molecules on the virus. LigoCyte then hopes to develop a drug or vaccine to block the sticking.

Biohazard

Anthrax


Barry Pyle has shown the Centers for Disease Control a method he and others developed to detect anthrax spores.

In 1996, when biological terror in the U.S. was still just a bad dream, a group of MSU scientists sought a grant through the Department of Defense (DEPSCoR) to develop a fast way of identifying potential agents. If biowarfare occurred, the MSU researchers wondered, how quickly could scientists pinpoint what organism was being used?

That grant bought a unique analyzing instrument made in France called a ChemScan. Sensitive enough to detect a single disease-causing microbe, the ChemScan was manufactured to detect bacteria in water. But the MSU microbiologists-Gordon McFeters, Barry Pyle, John Lisle and Susan Broadaway-began modifying the instrument methods supplied with the ChemScan system to detect microbes in other types of samples.

Ironically, they used a harmless cousin of the deadly anthrax for the project. The machine could automatically test a sample for the microbe in a few hours as apposed to the standard days-long method of growing microbes on Petri dishes.

And there the MSU research stopped. To test the ChemScan against the type of anthrax spores unleashed last fall in the U.S. postal service would have required biocontainment facilities that MSU doesn't have.

Last November, however, Pyle showed scientists at the Centers for Disease Control how to use a newer model of the machine for detecting anthrax spores. The agency had purchased one of the machines and plans to use it for research and to evaluate disinfection treatments such as the ones performed on congressional offices in Washington, D.C., said Pyle.

The MSU scientists will continue developing new ways of using the machine to detect other potential bioterrorist agents and will share the methods with the CDC.

Other anthrax-related research is occurring at LigoCyte. Director of molecular biology Susan Wimer-Mackin is developing anti-infective products to prevent the diseases associated with exposure to anthrax spores.

Annette Trinity-Stevens

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Anthrax Studies Hold Promise for Better Treatment

R. John Collier
Someone with ill intent will always be able to isolate the bacterium that causes anthrax, a researcher from Harvard Medical School said during a lecture at MSU.

"In most of the western United States, with a little bit of knowledge and some petri dishes, you can isolate Bacillus anthracis from the soil," said R. John Collier, the Maude and Lillian Presley Professor of Microbiology and Molecular Genetics from Harvard. "There's no way it will ever be eradicated from the soil."

The hope, though, is that the person who knows how to isolate the organism won't know how to turn it into a weapon, Collier added.

Collier is one of the foremost anthrax researchers, according to Mark Quinn, associate professor of veterinary molecular biology at MSU. Speaking to a packed lecture hall, Collier offered a new possibility for treating and preventing anthrax disease.

Anthrax is rarely found in humans, but it is much more common in plant-eating animals like cattle, sheep and bison, Collier continued. An animal's blood cells may house enormous numbers of the anthrax cells when it dies. As the animal decays, those cells form spores. The spores can live in the soil for years. Humans can become exposed to them through inhalation, absorption or open wounds, with inhalation being the most likely means to cause death.

"Clearly, we need better ways to treat it to reduce the mortality rate from what it was in this attack," Collier said, explaining that spores that enter the lungs germinate and spread infection to the bloodstream.

Vaccination is possible, but the military is the only large segment of the population that's vaccinated against anthrax, Collier said. The disease can be treated with antibiotics, but it doesn't take much to develop multiresistant strains.

Collier is hopeful about a third option that has evolved from his and other scientists" research into a three-part toxin. He said three ordinarily harmless proteins combine to create the toxin that helps anthrax infect its host. By intervening at critical times, scientists can prevent anthrax toxin assembly and possibly ward off its lethal effects.

Collier said the cycle offers many opportunities for intervention but he cautioned listeners about expecting a new treatment any time soon.

"An awful lot of work needs to be done to see if these are effective for treating animals and humans," he said.

Evelyn Boswell
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