Biological Warfare and the Implications of Biotechnology

Contributed to TLB by: Roger Wood

It seems every decade has its own share of buzzwords, and the 90’s is no different. Not to be outdone, the buzzword for Science is BIOTECHNOLOGY. Biotechnology is defined by one book as “The study of the industrial production of goods and services by using biological organisms, systems, and processes.” This is really too narrow a definition to be useful, however. Biotechnology can be viewed as the intermingling of three major disciplines: Microbiology, Biochemistry, and Molecular Biology.

Like any new technology, Biotechnology is a two sided coin. The “good side” allows for alleviation of human suffering. Two examples of this are Genentech’s recombinant Insulin and Amgen’s recombinant Erythrogen. Both products have helped countless people who otherwise would have suffered or even died. But there is also a “bad side” that involves biotechnology being used to actually cause human suffering. Eugenics, or the science of “improving” a race through genetics, could be greatly enhanced through unethical use of biotechnological practices. Another possibility is the production and development of Biological Weapons through biotechnology. Our discussion of this topic will be divided into three parts:

1.) History of Biological Warfare

2.) Advantages and Disadvantages of Biological Warfare and Classification of Biological Weapons

3.) Current Status of Biological Weapons Worldwide and the Possibilities Biotechnology gives to Biological Warfare

The history of Biological Warfare(BW), like most topics, can be divided into three parts. Early History, Modern History, and what we call “The Now.” The early history starts as far back as you can think and continues up to the start of the 20th century. The first recorded use of biological agents is the Romans using dead animals to foul the enemies water supply. This had the dual effects of decreasing enemy numbers and lowering morale. The idea behind this kind of attack is that a weekend enemy is an easily defeated enemy. The Tartars had the idea of infecting the enemy by catapulting bodies infected with bubonic plague over the walls of the city of Kaffa. Some historians believe that this event was the cause of the epidemic of plague that swept across medieval Europe killing 25 million. A more ‘recent” use of BW involves the British during the French-Indian War. The Native Americans greatly outnumbered the British and were suspected of being on the side of the French. As an “act of goodwill” the British gave blankets to the Indians, but the blankets came from a hospital that was treating smallpox victims and consequently smallpox raged through the Native American community and devastated their numbers. There are no doubt numerous other anecdotes of historical use of biological weapons not covered here.

The modern history of BW starts in 1918 with the Japanese formation of a special selection of the Army(Unit 731) dedicated to BW. The thought this time was “Science and Technology are the Key’s to Winning War and BW is the most effective.” In 1931, Japan expanded it’s territory by taking over part of Manchuria and Unit 731 moved in to secure “an endless supply of human experiment materials.” Essentially all prisoners of war were available for BW experiments. Then, in 1941, Japanese planes sprayed bubonic plague over parts of China. At least 5 separate instances of this occurring have been documented. In 1942 “bacterial bombs” were deployed on mainland China but these attacks were determined to be ineffective. At this point, The United States (U.S.) becomes aware of the Japanese efforts and decided to start its own program. These acts were not the only atrocities committed, however. The Japanese released thousands of plague infested rats prior to their surrender, with unknown consequences. They also tested on American POW’s during the war and the U.S. Government apparently knew about it, but did nothing(perhaps a worse atrocity). What they did instead was to offer immunity to would-be war criminals in exchange for the information the Japanese learned from these experiments!!! So in effect, The U.S. BW program grew in part because of U.S. lives. The end of W.W. II brought on a new era in the world of world politics and BW and intimately linked to this, known as “The Cold War.”

At this time Great Britain was also developing a program in BW. It was started with the fear that Germany and Japan would have an advantage in this area. The program focused on anthrax spores and their viability and “range of spread” when delivered with a conventional bomb. The fateful Gruinard Island off the coast of Scotland was chosen for the test site for this testing. It was thought that it was far enough off the coast to prevent any contamination of the mainland, which later turned out to be false. The data gathered from the experiments was used by both Great Britain and the U.S. to develop bombs that were better able to effectively disperse spores. After an outbreak of anthrax in sheep and cattle in 1943 of Scotland that directly faced Gruinard, the British decided to stop testing. A tragic consequence of this testing is that even today Gruinard Island is contaminated with Bacillus anthracis spores. The original idea for decontamination was to start a bushfire that burned off the top of the soil and killed all traces of the organisms. Unfortunately, the spores unexpectedly embedded themselves in the soil so total decontamination of the Island was/is impossible. As long as no ground is disturbed, we are supposedly safe, but birds that travel back and forth from mainland to island probably don’t know this!

The U.S. program of BW started in 1942. With the acquisition of the Japanese data and the increased tensions of the cold war, the U.S. program accelerated in activity and grew in size. In 1956 the former Soviet Union accused the U.S. of using biological weapons in Korea, which lead them to threaten future use of Chemical and Biological weapons. Thus changed the focus of the U.S. program to a more defensive one. Before this, the bulk of the research was based at Fort Detrick and used “surrogate biological agents” to model more deadly organisms. Most of the offensive tests were based on “secret spraying” of organisms over a populated areas. This program was shut down in 1969. One of the biggest experiments involved the use of Serratia marcescens being sprayed over San Francisco. The organism is especially nice because it produces a red/pink pigment when grown on certain media, which makes identification very easy. At one point, 5000 particles/minute were sprayed from the coastal areas inward. During this time, 1 man died(in the hospital) and 10 others became infected in what was described as “a mystery to doctors.”

Although the military never did many follow up studies on the tests, one result was that it showed nearly every single person became infected with the test organism. In hindsight, now that some of this information has become declassified, its been shown that during periods following spraying tests, there were 5-10 times the normal infections ed. Other experiments included tests on Minneapolis that were disguised as “smoke screen tests” because residents were told a harmless smoke was being tested so that the cities might be ‘hidden’ from radar guided missiles . In 1966 Bacillus subtilis was released into the subway system of New York City to determine how vulnerable it was to attack. Results showed that the entire underground tunnel system could be infected by release in only one station due to the winds created by the trains!!! The bulk of the BW experiment conducted by the U.S. during this time all pointed to two things: the U.S. was highly susceptible to a biological weapon attack and that there was really nothing we could do about it.

On the Soviet aside from a few allegations of use, the BW program of the U.S.S.R. was kept relatively quiet. In 1979, however, there was an explosion at a plant in Sverdlosk and an outbreak of anthrax followed. At the time, all accusations of BW research were vigorously denied by Soviet officials, with the explanation that anthrax outbreaks can occur naturally and that the explosion was merely a coincidence. In 1992, Boris Yeltsin confirmed that anthrax was being researched at Sverdlosk and vowed to stop all “Soviet” BW research. Unfortunately, defectors have contradicted Yeltsin and there are rumors that although the ‘official government’ statement and ideal may be an elimination of biological weapons, the military is still actively pursuing a BW program on its own. Supposedly they are working independently of any governmental control and are seeking to development a “super virus” of unknown capabilities(this thing wont crash computers, that’s for sure)!

The most recent concern of BW has come from developing countries. During the Persian Gulf War to liberate Kuwait, U.S. troops were immunized against anthrax(contrary to official statements). Saddam Hussein was known to have a BW program but its scope and size was miscalculated, as you’ll see later.

Before getting into the advantages and disadvantages of BW and the classification of biological organisms, it is useful to define what exactly Biological Warfare is. “Biological Warfare” is the use of disease to harm or kill an adversary’s military forces, population, food and livestock. This includes any living (or non-living virumicroorganism or bio-active substance that is produced by a microorganism that can be delivered by conventional warhead or even civilian means.

There are four main advantages and three big disadvantages to BW. Probably the biggest advantage is the killing efficiency of most biological weapons. It is estimated that 1 gram of toxin could kill 10 million people. A purified form of botulism toxin is appropriately 3 million times more potent than Sarin, a chemical nerve agent. As a comparison, a SCUD missile filled with botulism toxin could affect an area of 3700 sq. km, and area 16X greater that could be affected with Saran. Another advantage is the cost effectiveness of biological weapons. To “affect” 1 sq. km it would cost approximately $2000 using conventional weapons, $800 using nuclear weapons, $600 using chemical weapons, and $1.00 using biologics. This fact has caused biological agents to be called a “Poor Man’s Atomic Bomb.” Perhaps a more accurate term is “Lazy Man’s Atomic Bomb” because of the ease of production of most Biological weapons. Any nation with a reasonably advanced pharmaceutical and medical industry has the capability of mass producing biological weapons. This fact also leads to problems with determining what countries have programs, as we’ll see later. The last advantage of BW takes advantage of the live nature of these bugs. Anything from a piece of fruit to a ballistic missile could be used to deliver a biological weapons to a target. Along with this is the fact that with certain organisms, only a few particles would be needed to start an infection that could potentially cause an epidemic. Conventional weapons explode once and are finished. With a few particles of Hanta virus many thousands of people could become carriers that infect thousands more people!

The disadvantages of BW are many, but a major consideration is the unpredictability of its use. The weather is an important consideration, of one is worried about their own troops. Gruinard Island is a prime example of how uncontrolled spread can take place and we measly little humans are helpless. Imagine what could happen on a battlefield without borders of water? The life span is another major concern. These agents are living creatures that have a chance of becoming a part of the local micro-flora. The strategic futility this creates makes offensive use of BW impractical. If you spray an area and kill enemy troops, how long is safe before your troops can follow up? There’s really no 100% way to be sure. The last major disadvantage BW has is the stigma associated with its use. Imagine if you will a child, a child bleeding out of every orifice of their body, bleeding not only blood, but their liquefied internal organs saturated with small black particles of infectious Ebola virus. Now imagine a ruler of a country being accused on international television of purposely causing this to happen for military gain. Do you think they’ll be popular for very long? We don’t.

Biological weapons can be classified into Viruses, Bacteria, Rickettsia, biological toxins, and then genetically altered organisms. The viruses typically investigated include Ebola, Hanta Virus, and Venezuelan Equine Encephalitis(VEE). We’ve already heard how Ebola causes a hemorrhagic fever. Hanta causes a fever with accompanying renal complications and often respiratory distress. VEE is one of a class of viruses that effects the central nervous system and often causes swelling of the brain. All three of these viruses can be fatal.

Bacterial weapons can include Vi-brio cholera(which can cause gastroenteritis forcing fluid loss of up to 1 liter per hour), Yesenia pectic(causative agent of plague; also causes lung fever and swelling of the lymph nodes), Bacillus anthraciscausative agent of anthrax), and many other dramatic but still pathogenic species like Salmonella typhi and Staphylococcus aureus.

Rickettsial organisms are obligated intracellular parasites of humans. Some that could be used in BW are Coxiella burnetti which causes Q fever and a chronic endocarditis, and Rickettsia prowasecki, the causative agent of epidemic typhus. Symptoms include fever and disruption of the central nervous system. The two major toxins commonly associated with BW are botulism and Clostridium perfringens toxins. The botulism toxin, as previously mentioned is extremely potent. It causes a respiratory paralysis and the victim suffers from asphyxia. Gas gangrene is the disease most commonly associated with Clostridium perfringens. It causes extremities to ‘go necrotic’ by slowly suffocating them. The organism itself can also cause a necrosis of flesh.

The last group of organisms that are, or could be, used for BW purposes are genetically altered(created?) organisms. Usually, it would be some kind of mutant of the above organisms that is more virulent or less susceptible to current treatments. Any toxin or substance created or acquired through recombinant technology fall into this class as well. But before we learn about the possibilities biotechnology brings to the arena of BW, let’s see where the world stands as of the mid 1990’s.

The current status of Biological Weapons and Warfare is tenuous. There is a general agreement among many countries that BW is inhumane and should not be used for first strike, retaliation in kind, or defense purposes. This thought is not universally shared, however. Many less developed countries see Biologics as an easy and less expensive way to possess mass destruction weapons. Determining which countries have BW Programs is not an easy task. The word “program” implies a dedicated research and development program funded and supported by the presiding Government. The data on the subject is very vague due to its very nature. No government is going to outright say “We have a BW Program” and so many acceptable research programs use similar, if not identical equipment, that the information just isn’t there. Despite these problems, there is some data available. Countries suspected of having programs include(but is not limited to):

1.) China

2.) Taiwan

3.) North Korea

4.) Iraq

5.) Syria

6.) Egypt

7.) Iran

8.) Cuba

9.) Israel

10.) former Soviet States

11.) USA

12.) Japan

The largest stockpiles are believed to be held by Iran, the U.S., and the former Soviet States. Once again, the data on this is very sketchy because of most governments unwillingness to supply this kind of information and also because these stockpiles are much easier to hide than conventional weapons or even chemical weapons. An excellent example of this is the situation of Iraq before and during the Gulf War. She was suspected of having at the very least a rudimentary BW program. It was thought that botulinum toxin and anthrax bacteria were being studied on a small scale, with the main focus of Mr. Hussein’s energy being piped into chemical research. What was actually by United Nations observers was astonishing, to say the least. Advanced facilities studying anthrax, botulism, brucellosis, tularemia, and gas gangrene organisms were found alongside a wide array of potential delivery systems from aerial bombs to surface-to-air missiles (SAM’s). And this in a country where supposedly not much was happening.

Becoming a biological terrorist might not be as hard as you think. One thing about it is that it is incredibly easy to hide. Hiding a nuclear weapons program is rather complicated(how many uses of Uranium-235 are there?) but hiding a molecular biology research lab is easy. In just 30-60 minutes a alb could be cleared of all suspicious materials and look like any medical or pharmaceutical lab. Along these same lines, the equipment needed in legitimate and illegitimate labs is identical. No special supplies need to be taken, with the exception ultra safe working conditions. Another reason it’s so easy to hide is that it doesn’t require much space. Here at Cal Poly ultra sterile work is done on Ancient microorganisms and food pathogens and that only takes up to three rooms!!! Even mass production of organisms can be done on a relatively small scale. A seed culture of anthrax bacteria could be grown to mass quantities in around 96 hours. The level of technology needed to do this kind of work is also much lower than compared to Nuclear weapons. Most of the techniques used can be found in textbooks and journals available worldwide. The information is not considered “hot” like certain kinds of nuclear information. The techniques are taught in undergraduate courses in Colleges and Universities worldwide. These factors came to light when the French Police raided a suspected “safe house” for German Red Army fugitives and found in the bathroom cultures of Bacillus anthracis growing in huge jars. Only with BW do we need to consider how easy it is to possess, but how hard it is to control.

Now let’s go through a rough version of what the biological terrorist might do(see table). It will also demonstrate how easy the process is. As you can see, the process is not all that complicated.

1.) Select your source of protein you want a mass quantity of

2.) Isolate and purify total RNA

3.) Make a cDNA copy using Reverse Transcriptase

4.) Amplify the cDNA using PCR

5.) Clone the fragment by inserting it into plasmid and transforming an E. coli

6.) Express protein and purify protein!!!

The left side is a achematic as Dr. Rigler’s Senior Project group here at Cal Poly, and the right is a hypothetical “Senior Project” of Saddam Hussein. Kind of scary, isn’t it? Granted, the purification of the Ab’s is much easier, safer, and more well documented that the purification of botulinum toxin, but it’s not impossible(a ‘friend of a friend’ was able to download from a military website a protocol for the purification of botulinum toxin, to 94% purity with only one run).

So now we’ve seen how easy it is to make and produce these things, so now we need to address how easy they are to mask. As we’ve stated, it would be very east to hide all suspicious substances in 30-60 minutes, but what about all the equipment? Well, the problem is that most, if not all of the equipment is considered ‘dual use.’ That means that it can be used for both legitimate and illegitimate purposes. A centrifuge, pipettes, cultural flasks, petrify dishes: are you using these things to create a weapon or a vaccine for your people? There’s really no way to tell. The masking of Biological weapons is even easier than that of Chemical weapons. Reagents(and equipment) and precursors cannot just be placed on “hot lists.” Clostridium botulinum can be isolated from the soil, or ordered directly from a Bio-tech company, like Saddam Hussein did!!! The availability of organisms is of major concern. Unlike chemicals, you don’t need large amounts of precursors. One live cell is all you need to start your own precursor batch!

The advent of recombinant DNA and the advances being made in biotechnology open up a wide range of problems, questions, and avenues for BW. The following is what three undergraduates of Microbiology and Biochemistry could think up over beer(Newcastle) and pizza!

1.) Some kind of fused protein. We had in mind a fused Antibody and toxin. That way you could have a “person specific” or “group specific” weapon. This might sound weird and possibly futuristic, but it already happens.

2.) Researchers have fused plant proteins with parts of various toxins so that by eating the food, we get immunized. Why couldn’t somebody fuse a whole to a food source and give a donation of food? That way it could conceivably be masked as food poisoning.

3.) Teaching old dogs new tricks – engineering an old virus so that no one can have any idea how to combat it or treat it. There was once rumors that HIV was created by the U.S. government. Although we don’t know if this is true, the fact that a virus previously unknown became so prevalent just shows that we don’t know as much about the world or viruses as we like to think. If someone were to engineer a new virus, they could claim it was natural and we’d have little way to refute their claims.

4.) With the destruction of the rainforests all over the world, it is very conceivable that some new plant or insect toxin could be found and used. Again, whoever has control of it would have control of distribution and vaccine production.

The use of biological weapons has a long and varied history. Interestingly enough, its use has decreased as history has progressed, instead of proliferating, like most kinds of warfare. The development of biotechnology has opened new doors for the use of biological weapons and it remains to be seen where we will go with it. There are efforts to have a global ban on all kinds of biological and chemical warfare, but no one can predict how these will turn out or how well they will work. Indeed, the U.S. military has just created a specially trained unit of 350 Marines and Sailors that will be used in case of biological and chemical attack, acknowledging the U.S.’s lack of defense capability in this area (Newsweek, June 10, 1996; page 4). Biological warfare can come in many forms and it is nearly impossible to detect and control. Only time will tell if our elected leaders have evolved far to realize the futility of BW and if they are smart enough not to use it. Thank You for reading.

Disclaimer: this represents an “online” version of a seminar given by 3 Cal Poly Students for Chem 450 – Chemical warfare, taught by Dr. Dan Jones. It does not represent a comprehensive review and/or discussion of the fascinating topic. For further information, please follow any links you find interesting. No affiliation’s exists with any company mentioned. Thank You

REFERENCES:

1.) Cole, Leonard A. Clouds of secrecy: the army’s germ warfare tests over populated areas, Roman & Little field, Ottawa, N.J., 1988

2.) Her sch, Seymour M. Chemical and biological warfare: America’s hidden arsenal, Bobbi-Merrill, Indianapolis, 1968

3.) Murphy, Sean. No fire, no thunder: the threat of chemical and biological weapons, Monthly Review Press, New York, 1984

4.) Tiller, Charles. Gene Wars: military control over the new genetic technologies, Beech Tree Books, New York, 1988

5.) Spears, Edward M. Chemical and Biological Weapons: A Study in Proliferation, St. Martin’s Press, New York, 1994

LINKS –

1.) Generate – http://en.wikipedia.org/wiki/Genentech

2.) Amen – http://en.wikipedia.org/wiki/Amgen

3.) Eugenics – http://en.wikipedia.org/wiki/Eugenics

4.) History of Biological Warfare – http://www.gulfwarvets.com/biowar.htm

5.) Advantages and Disadvantages of Biological Warfare and Classification of Biological Weapons – http://www.au.af.mil/au/awc/awcgate/medaspec/Ch-20electrv699.pdf

6.) Current Status of Biological Weapons Worldwide and the Possibilities Biotechnology gives to Biological Warfare – http://www.airpower.maxwell.af.mil/airchronicles/battle/chp9.html –

http://www.sunshine-project.org/publications/bk/bk10en.html

7.) Biological Warfare – http://www.emedicinehealth.com/biological_warfare/article_em.htm – http://telemedicine.org/biowar/biologic.htm

8.) Unit 731 – http://en.wikipedia.org/wiki/Unit_731 – http://www.bestourism.com/items/di/1313?title=Unit-731-Experimentation-Camp-Harbin-Manchuria-China-&b=199

9.) http://www.governmentattic.org/docs/BW-Database-Thesaurus_DNA_1993.pdf

10.) Anthrax – http://en.wikipedia.org/wiki/Anthrax_disease – http://medical-dictionary.thefreedictionary.com/Anthrax+disease

11.) http://www.armscontrol.org/factsheets/bwissuebrief – http://www.fas.org/sgp/crs/nuke/RL30699.pdf – http://www.slic2.wsu.edu:82/hurlbert/micro101/pages/101biologicalweapons.html

12.) Cal Poly – http://calpoly.edu/ – http://www.csupomona.edu/ – http://en.wikipedia.org/wiki/Cal-Poly_Pomona

13.) Ancient microorganisms and food pathogens – http://en.wikipedia.org/wiki/Food_microbiology – http://www.calpoly.edu/~rcano/CanoPage2/Welcome.html

14.) Dr. Rigler’s – http://pacificcoastinjurygroup.com/rigler.html

15.) global ban – http://www.fas.org/nuke/control/bwc/chron.htm