Jane's Defense Weekly, May 1, 1998 [Copyright 1998 Jane's Information Group Limited, All Rights Reserved]

Molecular bio-technology will transform agriculture, energy production, health care, and microelectronics, however, it will also pose significant military and strategic challenges. Whereas recombinant DNA technology offers great benefits to humankind, it also has a darker side - the genetic engineering of microbial pathogens, toxins, and even natural brain chemicals, to create more deadly and persistent weapons of war.

During the Cold War a consensus existed that the Soviet Union was using recombinant DNA technology for military purposes. It was also attempting the recombination of the venom-producing genes from cobras and scorpions (and even bees) with the DNA of normally harmless bacteria. Such an organism would infect the body and surreptitiously produce paralytic cobra toxin. If delivered as a respirable aerosol, such an engineered agent could infect tens of thousands of people.

Former Soviet scientists have described a jointly-operated gene warfare program between the military and the Soviet Academy of Sciences which enjoyed the full support of the Kremlin leadership. The United States, meanwhile, had unilaterally stopped military research into offensive biological and toxin warfare capability in 1972.

Some observers are worried that work in this field is continuing. However, Prof. Matthew Meselson, professor of biochemistry at Harvard University and former chairman of the Federation of American Scientists, has spent a great deal of time investigating what the Soviets, and now the Russians, are doing in the area of biological warfare. He maintains that Moscow's biological research programs are now minimal, pointing to the fact that at every one of the known BW installations, recent satellite imaging has not revealed any ultraviolet emissions.

However, biological issues continue to feature in the media. UK and US troops were inoculated against anthrax during their recent deployment in the Persian Gulf, and the UK government issued an all-ports alert following intelligence reports of an attempted smuggling of an anthrax toxin into the country concealed in duty-free merchandise.

Iraq's research into anthrax, botulinum toxin, gas gangrene bacteria, and various chemical poisons such as VX gas, is regarded as the greatest threat. Mustard gas and nerve agents were used in combat against Iran during the 1980-88 Gulf War, and nerve agents were deployed against a Kurdish village in 1988 with casualty figures in the thousands. Moreover, a little-publicized CIA report from 1996 - Intelligence Related to Gulf War Illness - said that: "There are no indications that any biological agent was destroyed by Coalition bombing."

Understanding the potential of biological warfare remains something of a gray area. Eric Croddy in his book Chemical and Biological Warfare, An Annotated Bibliography makes a persuasive case that it is actually quite difficult to kill huge numbers of people using chemical or biological warfare (CBW) agents. "In fact," says Croddy, "it is a considerable challenge to use microbes and biological toxins as weapons of any scale."

Most bacteria and protein toxins are fragile. They are thermolabile (unstable when heated), and are sensitive to acidic solutions and ultraviolet radiation from the sun. Not only must a biological agent 'jump' a number of environmental hurdles, it must also face the antibody/antigen response once inside human tissue.

"The Russians, as with ourselves in the West," says Croddy, "discovered a long time ago that the human body is remarkably resilient. The largest organ, the skin, is a selectively permeable shield against common bacteria, rickettsia and parasites that are ubiquitous in our atmosphere. Even if ingested in food or water, most microbes that would otherwise be remarkably virulent, usually die - their protein toxins denatured by acidic and enzymic action in the gut. And if they find their way in through a cut in the skin, likely as not almost all bacteria will be engulfed by the phagocytic guardians of our immune systems."

Overcoming these defensive mechanisms requires unique features: because the most 'hospitable' and vulnerable sites of entry are the lungs, any biological warfare microbes have to be delivered in some form of respirable aerosol to cause mass casualties. Consequently, microencapsulation is a possible future technology for delivery of some of the more fragile viruses.

Only the most hardy microbes survive the necessary processing in today's biological weapons, and one pathogen - anthrax - fits most criteria as an effective BW. In addition to its ability to form an aerosol, this bacterium on infection attacks the body's own defenses. Significantly, Russia has experimented with an anthrax strain that shows resistance to antibiotic treatment.

Found in domestic livestock, anthrax (Bacillus anthracis) is most commonly encountered among sheep. Shearing of these animals allows the bacteria to become airborne and is the causative agent in woolsorter's disease (a form of pneumonia).

As a spore-forming bacterium, anthrax can survive for decades in soil. This makes it ideal for freeze-drying into an exceptionally fine powder. Once ensconced in the lungs, the capsule surface of the spore resists the body's immunological response. One of three toxins released by the anthrax bacteria further reduces the body's ability to react protectively.

For decades, scientists believed that anthrax killed its victims by forming 'logjams' in the blood stream. Research has since determined that Anthrax toxin III (in combination with other factors) is arguably the most intrusive culprit. Multiplying in the lungs and then in the bloodstream, anthrax reproduces in ever greater numbers by geometric progression. Anecdotal reports of patients succumbing suddenly to anthrax following two or three days of symptoms are consistent with the release of lethal toxin.

Further investigations into anthrax reveal more unsettling discoveries, not least that some strains appear to be resistant to penicillin. This presents another dilemma: in the event of a terrorist anthrax attack, sufficient antibiotics for large population concentrations might not be immediately available. It takes time to manufacture antibiotics. If the threat is real, tens of millions of people would need to be immediately inoculated. Then there is the question of time and whether a therapeutic course could be given quickly enough.

The potential for genetic engineering of this bacterium also presents some horrific consequences: a bacteriological weapon, already well-suited to killing thousands of people within days, could be modified (as Russia has done) to make it resistant to antibiotics. It is, as one observer noted, the perfect weapon and there is every indication that Iraq possesses it in abundance. During the 1990-91 Gulf War, Iraq had large quantities of anthrax, with the intention of dispersing the bacteria over Coalition lines and across Israel.

An outbreak of anthrax in humans occurred in 1979 at Sverdlovsk (now Ekaterinburg) in the former Soviet Union. The original release of spores came from a biological warfare research laboratory in the southern suburbs of the city of 1.2 million people on the eastern slopes of the Urals. It was freeze-dried and the amount released into the atmosphere, according to Meselson, was anything from 4mg to 1g. (It is impossible to see 4mg with the naked eye.)

The resultant epidemic - 96 people were infected and 64 died - provoked intense international debate. The Russians never revealed how much anthrax was involved, and it took years to get to the point where they were prepared even to admit that it was an accident.

Speculation continues as to whether the accidental release resulted from activities prohibited by the Biological and Toxin Weapons Convention of 1972. Subsequent research has shown that under such circumstances, the fatality rate - without aggressive medical treatment - would have been more than 95 per cent.

Anthrax particles are most effective when they are within a certain micron range; the accepted wisdom indicates anything between 1-10m. Extensive research that has been carried out at the United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Maryland, has indicated that between 8,000 and 10,000 spores constitute an LD50 (lethal dose that will kill half of those infected) for humans. If anthrax is disseminated in sufficiently fine particles, it quickly gets into the bloodstream through the lungs.

Significantly, capsulating bacteria like Bacillus anthracis tend to defeat the body's immune system in two ways: first they resist being engulfed by white blood cells; second, they produce toxins that actually vitiate the human immune response. Then the bacteria start multiplying at a rapid rate, producing a most lethal toxin. Fortunately this symptom can be treated if caught early enough.

However, if the strain is the latest Russian development and proves to be impervious to antibiotics, any such treatment is rendered ineffective. The progression in such a case is simple; as the bacteria multiply, the level of toxin dramatically increases. Death can be sudden, following a few days of incubation. At Sverdlovsk the first victim was dead in days, and the last case died six weeks after infection.

The consensus is that anyone exposed to at least 20 anthrax spores per cubic liter of air for about 30min will probably receive close to an LD50 dose. If enough infected air is inhaled in that time, sufficient spores will enter the victim's lungs to make for an infectious dose.

Iraq carried out research in this field in laboratories at Al Hakam and Salman Pak, both on the outskirts of Baghdad. Although Coalition bombing in 1991 destroyed much of this, intelligence sources indicate that a significant measure of this BW program has survived intact. United Nations inspectors with the UN Special Commission (UNSCOM) are now searching for these assets. There is no doubt that the weapons exist - the problem is finding them.

Those countries with advanced research BW programs - such as Iraq, Iran, Libya, North Korea, and Syria - have anthrax occurring naturally, usually in such places as stockyards, wool shearing depots and the like. Governments are consequently able to seek scientific aid regarding treatment from humanitarian organizations as part of their ongoing research, because anthrax is a viable threat to livestock.

In 1986, before the breakdown of relations between Iraq and the US, the latter country supplied seed cultures of anthrax from the American Type Culture Collection, a laboratory in Rockville, Maryland. Spore samples were ordered by telephone and were sent, as a matter of course, by normal mail. Iraq and other renegade states working on BWs were then able to ask the relevant bodies for anthrax, claiming it was needed for research on antibiotic regimens.

This supply channel has since been blocked. According to the US Department of Defense (DoD), this is the most common method of creating a national BW program; as such, anthrax will continue to be the prime experimental pathogen, which is why it is regarded as such a serious threat.

Similarly, botulinum toxin - another 'pathogen of choice' for some developing countries - presents a different type of threat. In humans the toxin interrupts conduction between our peripheral nervous system and muscle receptors. Botox, as it is commonly called, is prescribed by doctors to treat Strabismus dystonius and other neurological disorders. It is also in everyday cosmetic use (to counter the onset of facial or other bodily wrinkles); botox makes the muscles flaccid or paralyzes them and they take months to recover.

If this occurred on a larger scale in the human body - where the majority of functions are determined by the way muscles behave - it would be fatal. It takes only an extremely small dose of botulinum to kill by respiratory paralysis. One such example is that of a woman who was poisoned by botulinum from eating just half of a green bean (which became infected during the canning process); there are not many microbes in half a green bean, but it contained sufficient toxin for it to kill. In the case of botulinism, the LD50 is about 0.001g/kg of body weight.

It is difficult to establish which of the two - anthrax or botulinum - is the more toxic. Certainly, botulinum acts considerably faster than anthrax which takes some days to get into the system before a septicaemic reaction is manifested. It can be preceded by flu-like symptoms. Although scientists are aware of the end result, not enough is known about botulinum to describe the sequence of events in detail.

For all this, neither anthrax nor botulinum has been tested on any scale in modern warfare. The Japanese Aum Shinrikyo cult made an unsuccessful attempt to rig a fan in a Tokyo building in order to spray anthrax spores over the city, but errors were made either in production or delivery. Militarily, the consensus is that these biological and toxin agents, while devastating to those who are unprepared, will not make or break wars. These pathogens remain unproven entities, but that does not make them any less potent as killers.

Aum Shinrikyo had access to a considerable amount of money (estimated at between US$300 million and US$1 billion) with which it hired chemists and biologists. Members of the cult were also reported to have arrived in Kikwit in the former Zare (now the Republic of Congo) at the time of an outbreak of the Ebola epidemic, to obtain samples for BW purposes.

Dr Jane Alexander, of the US Defense Advanced Research Projects Agency (DARPA), has studied the Japanese sect. She discovered that Aum had also undertaken research into the 0157:H7 variant of E.coli, which produces a toxin. Although E.coli is difficult to manipulate as an aerosol for BW use, such bugs can cause serious problems (in Osaka, Japan, 10,000 people were infected with a deviant form of E.coli 0157:H7).

What Aum was working on entailed the insertion of botulinum toxin inside E.coli bacteria in an attempt to manufacture a lethal carrier of this toxin. It is likely that Aum was investigating plasmids (small rings of DNA material that carry genetic information) to implant material into bacteria as well. In fact, pharmaceutical companies use E.coli plasmids to produce Vitamin C which is cheaper than trying to synthesize it - for these purposes E.coli is relatively innocuous and exists as a normal and healthy bacteria in human intestines.

In this way, says Croddy, the processing of E.coli as a weapon is an attempt to 'sneak' an E.coli strain into the body, which does not (or cannot) react immediately. This virulent new strain would then start to multiply. (It is much more serious if a toxin-generating gene has been inserted into a bacterial DNA.)

The ramifications of such genetic engineering are endless: in the case of the E.coli 0157:H7 strain, a toxin would be produced that can result in serious problems in children and the elderly, including renal failure. This strain of E-coli was responsible for an outbreak of food poisoning in Scotland during 1996, when more than a dozen people died and hundreds were taken ill.

Such effects can be taken one step further by inserting other genes to produce more virulent toxins, which is what the former Soviet Union was researching. Western intelligence agencies are aware that some work into biological warfare programs continued in the Commonwealth of Independent States until at least 1992 (the Sverdlovsk-17 BW plant was shut down in the same year).

The UK and US have monitored these developments carefully. They are in the process of determining the long-term implications (see IDR 4/1998, pp21-24), largely to ensure that the West has adequate defenses should the deployment of germ weapons become a reality.

One of the issues raised - particularly in the light of revelations about Iraq's BW program - is whether the US will ever again acquire a real offensive CBW capability. The consensus is 'absolutely not'. There are, however, numerous institutions - the military (USAMRIID in particular), universities, and so on - that spend a great deal of money in establishing how and which microbial pathogens might be manipulated, in order to anticipate what potentially hostile states or subversive groups might be working on.

DARPA is making a serious efforts to look closely at E.coli in terms of biological defense. Here, too, Russia has been working to transfer segments of genetic material from one bacterium to another, which is relevant in terms of antibiotic resistance. The upshot is whether or not a new germ manufactured by a hostile power can be effectively countered.

This has caused scientists to speculate how E.coli 0157:H7 originally came into existence, and how it became a toxin- generating microbe. There are some scientists who argue that the strain emerged naturally and mutated from bacteria exchanging genetic information. According to others, it could have been bio-engineered in a laboratory and spread from there. Most scientific favor rests with the first argument. In terms of research into E.coli, Croddy's view is that: "All the major European powers are tinkering about with it defensively. Personally, I would say that USAMRIID is further ahead than just about anybody else."

In the area of biological warfare defense, money - and a lot of it - is being spent. In the US, DARPA is co-ordinating most of the purse strings, channeling much of the money through the Biological Defense Program Office. Significant funds are also being diverted to educational institutions such as Johns Hopkins University and others in the academic research field.

Research into a small mass-spectrometer that will fit into a standard-sized briefcase which could conceivably be used under battlefield conditions is also under way. This will be capable of identifying in real-time if a friendly force is deployed in a 'hot' (contaminated) zone. Although this development is better suited for the characterization of chemical agents, it is hoped that highly accurate readings of the primary eight BW threats will also be available.

"Then," says Croddy, "if you detect any sort of biological presence, you may not know immediately what it is and whether it is a potential threat. But you will have a pretty good idea. You will probably want to 'suit-up' anyway."

The DoD concluded recently that the biological warfare threat was one area in which the United States has found itself to be the most vulnerable. This was said repeatedly at a symposium on the subject held in Atlanta, Georgia, in March. More than 2,000 delegates from 70 countries were present, many of them military officers. This indicates the significant level of interest in a menace which could, if ever released from the confines of a laboratory, herald a global epidemic.