Sunday, October 12, 2014

From Animal to Mankind

     Many diseases are 'species-specific', affecting one type of animal and not others. There are some kinds of microbes, however, which have the ability to 'jump' the species barrier (called 'zoonoses').

     Usually, this involves spontaneous 'mixing' of animal and human molecular content of the microbe, creating a 'natural' hybrid variety. This is what is believed to have happened with the H5N1 'bird' flu epidemic in and possibly the outbreak of SARS in 2002-2004. Since the first H5N1 outbreak occurred in 1987, there has been an increasing number of H5N1 bird-to-human transmissions, leading to clinically severe and fatal human infections.
     The 2009 flu pandemic (swine flu) was the second of the two pandemics involving H1N1 influenza virus (the first being the Spanish flu pandemic of 1918 , when more than 20 million people died worldwide). The 2009 version of the virus appeared to be a new strain of H1N1 which resulted when a previous triple reassortment of bird, swine and human flu viruses further combined with a Eurasian pig flu virus (leading to the term swine flu).
The Spanish Flu

     Tularemia, caused by infection with the bacterium Francisella tularensis, has a relatively low mortality rate (5 percent) but the microorganism is one of the most infectious bacteria on the planet.
     In 1941, the Soviet Union reported 10,000 cases of the illness but during the German siege of Stalingrad of 1942 (only one year later), this number rose to 100,000 with the vast majority of these cases on the German side.
     Ken Alibek (Colonel Kanatjan Alibekov), a former Soviet bioweapons researcher claimed that this surge in infections was no accident but rather was the result of biological warfare. Alibek later developed a strain of vaccine-resistant tularemia for the Soviets, before defecting to the United States in 1992.
Siege of Stalingrad

     Francisella tularensis occurs naturally in many organisms but is especially prevalent in rodents, rabbits and hares. Humans infection typically occurs through contact with infected animals, infected insect bites, the consumption of contaminated foods or the inhalation of the bacteria as an aerosol.
     Symptoms of tularemia appear within 3 to 5 days and vary dependent on the method of infection (inhalation, bite, etc). Patients experience fever, chills, headache, diarrhea, muscle pain, joint pain, dry cough and progressive weakness. Pneumonia-like symptoms can also develop and, if untreated,can progress to respiratory failure, shock and death.
Colonel Kanatjan Alibekov (Ken Alibek)

     Tularemia does not transfer between human hosts and can be treated with antibiotics or prevented with a vaccine. It does spread very rapidly between animal hosts and humans, however or when used in aerosol form.
     Severe acute respiratory syndrome  (SARS) is a viral respiratory illness in humans caused by the  coronavirus.
     Between November 2002 and July 2003, an outbreak of SARS in Hong Kong spread rapidly across the globe, nearly becoming a pandemic (global epidemic), infecting 8,422 people and resulting in 916 deaths worldwide (mortality rate of 10.9%). Within weeks in early 2003, the SARS virus had spread from Hong Kong to 37 other countries.
     Initial symptoms of the disease resemble those of influenza and are non-specific such as muscle pain, fever, lethargy, cough and sore throat. The only symptom common to all patients appears to be a fever above 38 degrees Celsius. Chest X-ray findings may be normal in the beginning but later show signs of severe pneumonia.
SARS Coronavirus

     There is no specific treatment for SARS. Antibiotics, effective against bacteria are ineffective for this disease, as SARS is caused by a virus.
     Treatment of SARS is supportive with medication to counteract fever, supplemental oxygen and ventilation support if needed. SARS cases should be isolated, preferably in with complete barrier nursing precautions taken for any necessary contact with these patients.
     Initially anecdotal support for antiviral medication and steroids was recommended but there has been no published evidence to supported this therapy. Chinese researchers were reported to have produced a SARS vaccine in December 2004.
SARS Pneumonia

     Reports out of China on some recovered SARS patients showed severe long-time difficulties which persist, most typically pulmonary fibrosis and major depression.
     Chinese researchers were reported to have produced a SARS vaccine in December 2004.
     During the worst of the SARS outbreak in China, in the spring of 2003, a  SARS conspiracy theory emerged. Sergei Kolasnikov, a Russian researcher and member of the Russian Academy of Medical Sciences, publicized his claim that the SARS coronavirus was a synthesis of measles and mumps viruses.
     According to Kolasnikov, this combination could not be formed in the natural world and thus the SARS virus must have been produced under laboratory conditions (a man-made microbe). Another Russian scientist, Nikolai Filtov, chief of epidemiological services in Moscow, had also suggested the SARS virus was probably man-made.
Asian Palm Civet

     Many Chinese believed that the SARS virus was a biological weapon manufactured by the USA. The failure to verify the source of the SARS virus convinced many people that SARS had been artificially synthesised and spread by some individuals or governments.
     Circumstantial evidence later suggested that the SARS virus crossed over to humans from Asian Palm Civets ('civet cats), which are frequently often killed and eaten in Guandong, the area in southern China where SARS was first discovered.
     Some in Taiwan and the United States have also expressed doubts as to the civet cat source of the disease and have speculated that SARS could be a biological weapon developed by the mainland Chinese.
     Other scientists acknowledge the possibility that the SARS virus could indeed be man-made. Genetic material in the SARS virus DNA sequence does not match any previously known virus, implying it could have been synthesized.
Guandong Province, Southern China

     On the other hand, coronaviruses similar to SARS have been found in bats in China, suggesting bats may be the natural reservoir for this disease.
     Cultivated food crops (plant or animal) can be an effective target for a biological weapon. Deprived of food, populations weaken, panic, riot and eventually die.
     The USA and Russia have focused much deal research on diseases and insects that target key food crops.
     Rice blast is a crop disease caused by the fungus Pyricularia oryzae. The leaves of affected plants develop gray lesions composed of thousands of fungal spores which multiply and spread from plant to plant, draining the plants and leading to much lower crop production. Breeding resistant plants may be a defensive measure against some crop disease but with rice blast resistance would have to be bred against 219 different strains.
Rice Blast

     Deployment of such a bioweapon would lead to severe starvation as well as financial losses and other huge problems.
     Fusarium oxysporum (a mold) has been considered as a weapon for drug plant eradication.  This fungus, has devastated commercial crops such as bananas and muskmelon in the past and is under investigation for the potential to destroy coca and cannabis plants, from which cocaine and marijuana are derived.
     Host specificity for this mold is narrow and species 'jumping' is rare which suggests that targets plant species can be selected without posing danger to other commercial crops.
Fusarium oxysporum

     When Ghengis Khan and his 'mongol horde' invaded Europe in the 1200s, he let loose a devastating biological weapon. The steppe cattle used by his supply trains introduced a deadly cattle plague which became known throughout the world today by its German name, rinderpest.
     Rinderpest (cattle plague or steppe murrain) is caused by a measles-related virus (morbillivirus), affecting principally cattle as well as other ruminants such as goats, bison and giraffes.
     This highly contagious disease causes fever, loss of appetite, dysentery and inflammation of the mucus membranes. The condition continues for six to 10 days at which point, the animal typically succumbs to dehydration.
Ghengis Khan

     The spread of rinderpest has followed the changing character of human empires, migrations and invasions, reaching out to the corners of the globe. In Africa, outbreaks have, at times, been so severe that human beings became lions' prey and herdsmen committed suicide.
     The disease likely originated in Asia (Ghengis Khan) spreading with the transport of cattle. Other cattle disease outbreaks were recorded in ancient times, including a cattle plague as one of the ten plagues of Egypt described in the Hebrew Bible.
     Cattle plagues recurred throughout history, often following wars and military campaigns. They hit Europe in the 18th century, with three long pandemics 1709 and 1786.
     There was a major outbreak covering the whole of Britain in 1865/66. An outbreak in the 1890s killed 80 to 90% of all cattle in southern Africa. Sir Arnold Theiler was instrumental in developing a vaccine that curbed the epidemic. In 1982–1984, an epidemic of the disease in Africa cost at least an estimated $500 million in stock losses.
Steppe Cattle

     On 8 August 2011, the United Nations declared the disease eradicated, making rinderpest only the second disease in history (following smallpox) to be fully wiped out.
     The disease was mainly spread by direct contact and by drinking contaminated water but also by air. Initial symptoms included fever, loss of appetite, and nasal and eye discharges followed by erosions in the mouth, the lining of the nose, and the genital tract. Constipation followed by diarrhea, is also a common feature. Most animals are dead within 6-12 days of onset of clinical signs.
     Rinderpest was researched as potential biological weapon by the USA before that country halted its biological weapons program. Rinderpest was considered as a biological weapon because of its high rates of morbidity and mortality rate (lots of sick and dead animals), the disease's high and rapid rate of spread once introduced into non immune herds and because cattle herds in the US and other developed countries are not routinely immunized against rinderpest.
Effects of Rinderpest

     Many of the scariest bioweapons on the planet have their roots in the ancient world.  A few, however, are terrifyingly new.
     Nipah virus came to world attention in 1999 when an outbreak occurred in the Nipah region of Malaysia, infecting 265 individuals, killing 105. Ninety percent of those infected handled pigs for a living but health workers su­spect the virus naturally occurs in fruit bats.
     The virus may spread through close physical contact or contaminated body fluids but human-to-human transmission has not yet been reported.

     Those infected with Nipah suffer from 6 to 10 days with symptoms that range from mild, flu like conditions (fever, muscle pains) to encephalitis (inflammation of the brain). In the more severe cases, patients experience drowsiness, disorientation, convulsions and ultimately coma.
     The virus carries a mortality rate of 50 percent with no treatments or vaccinations. It is this high mortality rate which makes Nipah virus attractive as a biological weapon.
     Nature is continually providing the human species new ways to destroy itself. As the biological sciences have advanced, it has become apparent that natural, human diseases have not been enough and, for weapons, we needed to borrow from nature's arsenal of animal micro-organisms.
     * Animal Diseases in Man and Their Use as Biological Weapons: subject of research for the novel Vaccine - Amazon Kindle.

Wednesday, October 1, 2014

Changing the Recipe-Genetic Engineering and Biological Weapons

     Genetic engineering or genetic modification is a technique which involves the direct manipulation of an organism's genome using modern biotechnology. The process changes the genetic makeup of an organism, removing or adding to the organism's genetic material (DNA).
Genetic Engineering

     Artificial selection (also known as selective breeding) involves intentional breeding of animals or plants for certain traits, or combination of traits and has been carried out for thousands of years. This is different from Darwin's natural selection where the environment selects out or adds individual traits which may be needed for better survival. But artificial selection can also be unintentional. For example, it is believed that domestication of crops (such as certain grains) by early humans was largely unintentional, occurring by chance as the best seeds tended to survive or provide better nutrition.
     Mutagenesis is another process that can be used to change the genetic information of an organism. Alteration of a gene or group of genes may occur spontaneously in nature or can be a result of exposure to mutagens, chemicals or radiation which induce the changes in the nuclei of the cell where the DNA is held.
The Development of Maize-
Artificial Selection?

     It is a technique which can also be achieved using laboratory procedures, creating genetic changes in the organism. In nature, mutagenesis can lead to cancer and various heritable disease but it is also the driving force of evolution.
     Genetic engineering, mutagenesis and artificial selection are the three techniques used by biologists seeking to create the ultimate biological weapon, a weapon which has no cure, no vaccine to protect against it or even a weapon which can selectively attack only certain members of the human species.
     But the creation of a biological weapon does not require genetic engineering. There are many deadly 'natural' organisms which are readily available to those who are determined to obtain them (smallpox, anthrax). However, the expansion of modern biotechnology in the medical and pharmacological fields as well as means of production has led to an easy availability of knowledge and facilities.
     Many countries which had previously been able to access only rudimentary technology, can now access high-tech facilities for vaccine or organism production that could be easily be altered for the production of biological weapons.
     Today, nearly all countries have the technological potential to produce large amounts of pathogenic microorganisms safely. 'Classical' biological agents can often be made more efficiently today than their natural counterparts using even simple genetic techniques. With this modern biotechnology, it is possible to create completely new biological weapons.
     Not only can artificial organisms be created which are resistant to antibiotics and vaccines but these new organisms are also more toxic, harder to detect and more stable in the environment.

     The use of genetic engineering in the development of biological weapons is not merely a theoretical possibility. Genetic engineering has been used in the former Soviet Union, with the creation of the USSR's 'invisible anthrax' where Soviet scientists inserted an 'alien' gene into Bacillus anthracis, altering its immunological properties. Vaccines against this new strain of anthrax proved to be ineffective.
     Anthrax is a prime choice as a biological weapon since it ban be produced in large amounts, acts rapidly and remains robust in the environment, surviving years buried in the soil (see post: Biological Weapons - How Bad Can They Be?). Ideally, the disease must be treatable, or a vaccine must be available for one's own troops.  However, potential victims of an anthrax attack can be treated with antibiotics even several days after an infection and, in most circumstances, only a minority of the infected die from anthrax. But just a very simple genetic intervention, such as that carried out by the Soviet Union, is often enough to 'change the rules of the game'  producing (as in the Soviet case) a form , resistant to vaccine and possibly even a variant resistant to antibiotics.
     There are a number of different approaches to the development of a 'synthetic' biological weapon, that is a natural disease which has been genetically altered or'enhanced'.
State Research Center for
Applied Microbiology, Obolensk

     Development of symptom-altered biological agents is one method of choice. Researchers from the State Research Center for Applied Microbiology in Obolensk, (about 85 kilometers south of Moscow) inserted a gene into Francisella tularensis (the bactrium which causes tularemia), making the bacteria produce beta-endorphin, an endogenous human drug (see post: Your Personal Narcotic). This genetic alteration caused changes in the behaviour of mice infected with this transgenic bacteria.
     Supposedly, the endorphin gene was not introduced into a fully virulent strain, but only into a vaccine strain. If inserted into virulent F. tularensis, the victims would not have shown the usual symptoms of tularemia, but instead unusual symptoms which would, in theory at least, have obscured the diagnosis and delayed therapy.

     The transfer of a lethal factor to harmless human gut bacteria through genetic engineering could also transform previously harmless bacteria into lethal biological weapons. This was actually achieved by US researchers in 1986. They isolated the gene for the lethal factor of Bacillus anthracis (the causative agent of anthrax) and introduced into Escherichia coli (a normal and harmless resident of the human gut).
     It was reported that the lethal factor protein was active in E. coli and displayed the same deadly effects as it did when in its native B. anthracis.
     There have been a number of countries which have attempted to create antibiotic resistant anthrax and tularemia.
     German researchers at the Santitaetsakademie der Bundeswehr in Munich cultured genetically engineered Francisella tularensis subsp. holarctica bacteria, a close relative of the causative agent of tularaemia. An antibiotic resistance marker gene (tetracyclin) was been inserted into these bacteria.
Porton Down

     British researchers from Porton Down in the UK used genes conferring resistance to antibiotics for genetic studies in fully virulent strains of anthrax.
     In the 1980s, a researcher at the University of Massaschussetts introduced antibiotic resistance genes into anthrax, rendering it less treatable with antibiotics.
     Researchers from the Institut Pasteur in Paris did similar work as their counterparts in Russia, introducing antibiotic resistance genes into anthrax bacteria.

     Detection of biological weapons depends on molecular recognition of the microbe using antibodies similar to the human immune system. Altering the immunogenicity of the microorganism not only overcomes vaccinations but also the detection systems.
Institut Pasteur

     In 1997, the same Russian research group from Obolensk published a paper on another effort to genetically engineer anthrax.  The team was able to put new genes into fully pathogenic strains of anthrax and alter the bacteria's immunopathogenic properties, making existing anthrax vaccines ineffective against the new genetically-engineered types.
     The Russian researchers also constructed a new vaccine against the new strain which potentially enabled the army using such a weapon, vaccination of their own soldiers against a specific strain, while the enemy remained vulnerable.
State University of New York in Stony Brook
      In 2002, a research team at the State University of New York in Stony Brook chemically synthesized an artificial polio virus from scratch  beginning with the genetic sequence of the agent which was available online. They ordered small, tailor-made DNA sequences and combined them to reconstruct the complete viral genome. Then, the synthesized DNA was brought to life by adding a chemical cocktail that initiated the production of a living, pathogenic virus.
     Many countries (such as the US) have investigated the development of material-degrading microorganisms aimed at destroying fuel, construction material or stealth paints. Many natural microorganisms are able to degrade nearly every material and are already being used to detoxify environmental pollution. The natural organisms, however, are slow-acting and unreliable, but, with the help of genetic engineering, the development of much more effective organisms might become possible—probably effective enough to be used as biological weapons.

     These investigations have been sparked by renewed interest in non-lethal weapons for use in media-sensitive military operations so that visible civilian victims can be avoided.
     In 1998, the US Naval Research Laboratory in Washington DC was declared to be developing genetically engineered fungi with offensive biological warfare potential. Researchers were able to isolate natural microorganisms which degrade a variety of materials, such as plastics, rubber and metals and used genetic engineering to make them more powerful and focused. One of these genetically engineered microbes apparently can destroy military paints in 72 hours.
     James Campbell, principal investigator at the Naval Research Laboratory, noted possible applications of this technology including microbial derived or based esterases, capable of stripping signature-control coatings from aircraft, allowing easier identification of enemy rockets or airplanes.

US Naval Research Laboratory
     The USA also increased efforts to identify microorganisms that kill drug-producing crops and, by the late 1990s, this research was focused on two fungi. The organism, Pleospora papaveracea, a killer of the opium poppy, was conducted in Tashkent, Uzbekistan, with financial and scientific support from the USA.
     Pathogenic Fusarium oxysporum strains developed in the USA to kill coca plants were scheduled for field tests in Colombia in 2000. These two species of fungi provide an excellent example of the hostile use of biological agents. In Colombia, the biggest areas of coca and opium poppy cultivation are in combat zones, and the 'War on Drugs' is part of that country's continuing armed conflict.
Pleospora papaveracea

     Some claim that the pursuit of crop-killing fungi as weapons would be a further slide down a slippery slope that, by following the same logic, could easily lead to the use of other plant pathogens, animal pathogens or even non-lethal biological weapons against humans.
     Another example of new ideas in warfare concern substances which are not necessarily biological weapons but new types of chemical, or rather biochemical weapons.  This area came under the spotlight of the international media after the use of psychoactive substances in the Moscow hostage crisis (Nord-Ost Siege) in October 2002, causing the death of more than 170 people.
     The chemicals used by the Russian 'swat' team to free the hostages held by Chechen rebels were probably narcotic based but the authorities refused to divulge the exact nature of the chemicals used.
     Many of these supposedly 'non-lethal' chemical weapons had been developed as early as the 1950s, particularly a substance called 'BZ (3-quinuclidinyl benzilate)', known in the US army as 'sleeping gas' and more commonly as 'buzz' because of its abbreviation as well its effects on the mental state of its victims.
Nord-Ost Siege (October 2002)

     BZ, invented by Hoffman-Laroche (see post: The Dawn of the Drug Dealers) in 1951,  is an anticholinergic compound (similar to a neurotransmitter-see post: The Genetics of Drug Addiction) related to scopolamine, atropine and other similar chemicals. Dispersal could be carried out as an aerosolized solid for breathing in or as agent dissolved in one or more solvents for ingestion or absorption through the skin.
     In 1998, the British accused Iraq of having stockpiled large amounts of a substance called Agent 15, a glycolate anticholinergic incapacitating agent chemically either identical to BZ or closely related to it. Agent 15 was reportedly stockpiled in large quantities prior to and during the First Gulf War.
Project SHAD

     During the Cold War in the US, Project SHAD (Shipboard Hazard and Defense)  a project under the guise of the Department of Defense was a series of tests carried out to investigate the use of both chemical and biological weapons related to a program dubbed, Project 112. Also known as the Edgewood Arsenal experiments, Project 112 investigations were said to part of CIA mind-control programs started after WWII.
     The experiments were performed at the Edgewood Arsenal, northeast of Baltimore, Maryland, and involved the use of hallucinogens such THC, LSD, BZ and other chemical and biological agents.
     BZ is odorless, very stable in most solvents and has a half-life of 3-4 weeks in moist air. It is persistent in soil, water and on most surfaces. BZ can be synthesized in clandestine laboratories but its recreational use is almost nonexistent, because of its unpleasant effects.
     Because BZ is odorless and nonirritating with delayed symptoms for several hours after contact, the only immediate indications of its use may be the white smoke emanating from delivered weapons.
     There is little risk of absorption f BZ through the skin or contact hazards from aerosols that have settled out onto exposed surfaces. The most protective response is to don a protective mask with a good quality aerosol filter. There is the possibility, however, that BZ could be employed to render activity through skin by the addition of a skin penetrating solvent or by using a secondary aerosol, contaminating the surrounding terrain with BZ in bulk micro-pulverized form.
BZ (3-quinuclidinyl benzilate)

     BZ is distributed to most organs and biological tissues of the body and is able to cross the blood-brain barrier, conferring effects upon the central nervous system.
     BZ an incapacitating rather than a toxic warfare agent and acts as a competitive inhibitor (blocker) of the neurotransmitter acetylcholine neurons.  As the concentration of BZ at these sites increases, the proportion of receptors available for binding to real acetylcholine decreases and the end organ 'sees' less acetylcholine. This leads to clinical effects reflective of under-stimulation of end organs, such as blurry vision, dry mouth and skin, decreased sweating and dilated pupils. With decreased sweating, the body core temperature rises. Effects on smooth muscle include decreased bladder tone with possibly severe bladder distension.
     BZ typically raises the heart rate initially, but hours later, depending on the dose of BZ, the heart rate falls to normal or may become slow.
     Central nervous system effects are more profound and include a decrease in the level of consciousness, beginning with drowsiness and progressing through sedation to coma. The patient is often disoriented to time and place with disturbed abilities in judgment and insight. The patient may abandon socially imposed restraints and resort to vulgar and inappropriate behavior. Perceptual clues may no longer be readily interpretable, and the patient is easily distracted and may have memory loss, especially short-term memory.
     The victim may also experience including illusions (misidentification of real objects) and hallucinations (the perception of objects or attributes that have no objective reality). Another effect of BZ poisoning is behavioral lability, with patients swinging back and forth between quiet confusion and self-absorption in hallucinations, to frank combativeness. Moreover, as other symptoms begin to resolve, intermittent paranoia may appear. Automatic behaviors common during resolution include crawling or climbing motions, previously called 'progresso obstinato' in old texts describing dementia.
Mass Hysteria

     BZ can also produce 'shared' illusions and hallucinations (folie en famille, folie a deux, mass hysteria). An examples of this is a case where two individuals took turns smoking an imaginary cigarette clearly visible to both of them but to no one else and another case where two victims of BZ played tennis with imaginary rockets.
     The course from BZ poisoning takes place in four stages. Stage 1 begins within 30 minutes to 4 hours after exposure and is characterized by parasympathetic blockade (dryness, blurry vision, etc.).
     The second stage (4 to 20 hours after exposure) is characterized by stupor with loss of balance and increased body temperature.
     Stage 3 (20 to 96 hours after exposure) produces full-blown delirium.
     The fourth stage or resolution stage, is characterized by paranoia, deep sleep, reawakening, crawling or climbing automatisms, and eventual reorientation.
     But because BZ can cause very different effects in different individuals, it was considered to be unreliable, leading to the banishment of this substance from the US chemical arsenal in the late 1960s.
     More efficient classical biological warfare agents will probably have only a marginal role, even if the genetically engineered 'superbug' is still routinely featured in newspaper reports. More likely and more alarming are weapons for new types of conflicts and 'niche' warfare scenarios, such as or covert operations,  economic warfare or sabotage activities.
     Biological organisms can be modified in a variety of different ways. Toxins, for instance, can be produced by adding the DNA coding for its production to previously harmless bacteria. Advances in biotechnology have made it possible to synthesize certain viruses based on its genome (the organism's genetic instructions) and using basic materials such as DNA.
     Dr. Eckard Wimmer demonstrated this by re-creating the poliovirus in 2001, followed by Dr. Craig Venter's synthesis of the bacteriophage phiX174 in 2003 and the 2005 re-creation of the 1918 flu virus by Dr. Jeffrey Taubenberger and Dr. Terrence Tumpey.
Dr. Craig Venter

     In Greek and Roman mythology, the chimera combined certain parts of lion, goat and serpent into one monstrous form. In the late medieval age, artists often used the creature as a symbol to illustrate the complex nature of evil. Today, a chimeric organism is a life form which contains genes from a foreign species but the goal or the end result is not always 'evil'.
     But throughout human history, abuse of science almost always occurs. Geneticists have discovered the means to increase the lethality of such biological weapons as smallpox and anthrax by tweaking their genetic structure. By combining genes, however, Scientists have discussed creating a virus that could trigger two diseases at once.
     During the late 1980s, the Soviet Union's Chimera Project, according to former Soviet researcher, Ken Alibek (see post: From Animal to Mankind) studied the feasibility of combining smallpox and Ebola into one super virus.
The Mythological Chimera

     Other potential nightmares involve scenarios of stealth viruses, strains which require certain triggers to produce infection. The stealth virus would remain dormant for a period of time until triggered by predetermined stimuli.
     Other possible chimeric weapons might require two components to become effective, such as a strain of botulinum toxin that only becomes lethal when combined with the botulinum toxin antidote.
     Before the closure of the American offensive biological warfare program by President Nixon in 1969, offensive agents had already been developed at the United States Army's biological-warfare laboratories at Fort Detrick, Maryland. These products which included powdered spores and viruses, were loaded into bombs and other delivery systems stored at Pine Bluff, Arkansas. The 1969 budget for Chemical/Biological Warfare research was reported to be $300 million with $5 million for herbicides designed to kill food crops or strip trees of foliage to deprive enemy forces of ground cover.
     By the late 1960s, the United States had developed a biological arsenal which included numerous bacterial pathogens, toxins, and fungal plant pathogens that could be directed against crops to induce crop failure and famine.
The Korean War

     During the Korean War, the Soviet Union, China, and North Korea accused the United States of waging biological warfare against North Korea and China. However, no epidemiologic evidence was ever found to support the North Korean claim of having experienced epidemics.
     Numerous other unsubstantiated allegations were made during the Cold War, including a Soviet accusation that the United States had tested biological weapons against Canadian Eskimos, resulting in a plague epidemic, an accusation that the United States had planned to initiate a cholera epidemic in southeastern China as well as another Soviet accusation of a United States and Columbian biological attack on Columbian and Bolivian peasants.
     In anticipation of the 1972 Biological Weapons Convention, President Nixon terminated the United States offensive biological weapons program by executive order in 1969. From that time on, research efforts were directed exclusively to the development of defensive measures such as diagnostic tests, vaccines, and therapies for potential biological weapons threats. Stocks of pathogens and the entire biological arsenal were destroyed but small quantities of some pathogens were retained at Fort Detrick to test the efficacy of investigational preventive measures and therapies.

     The Cobra Event is a 1998 novel by Richard Preston in which a virus, called 'cobra' was created, mixing the incurable common cold virus with one of the smallpox virus. In this fictional story, the disease that resulted from the virus was called brainpox, a genetically-engineered recombinant virus which caused nightmares, fever, chills, runny nose, encephalitis (brain swelling), and herpes-like boils in the mouth and genitals.
     But 'brainpox' may not be that far from reality. Venezuelan equine encephalitis (VEE) is an alphavirus related to a number of different viral diseases including eastern equine encephalitis and western equine encephalitis. These viruses can be easily produced in large amounts and aerosolized (for biological weapons purposes).
     Considered as a biological agent, VEE viruses are relatively stable, and can potentially injury thousands. Most importantly, the VEE virus is susceptible to genetic manipulations, which may enhance its infectiousness and virulence for biological weapons purposes. VEE is a particularly appealing biological weapon since it only requires 10-100 pathogens to infect a person. Infection with VEE infections is rarely fatal but can cause severe symptoms similar to influenza and hence can be difficult to diagnose. These encephalitis fevers cause inflammation of the brain and long-term side effects such as nervous system damage. A potential biological attack using VEE virus could be through the aerosolized route, but would be most effective during periods when mosquitoes are most active.
Venezuelan Horse Deaths in Florida

     In April 2009, 3 vials of VEE were found to be missing from the Fort Detrick research facility and soon after, 21 thoroughbred horses from a Venezuelan polo team competing in Florida suddenly dropped dead.
     The United States weaponized VEE as an offensive incapacitating agent before the termination of its biological weapons program in 1969 and the Soviet Union also harnessed VEE as a biological weapon.
     According to Ken Alibek (see post: Biological Weapons in the Twentieth Century), Soviet scientists at the All-Union Scientific-Research Institute of Molecular Biology in Koltsovo, Siberia experimented with splicing VEE genome into smallpox viruses, creating a recombinant smallpox-VEE chimera virus that resembled smallpox under a microscope but produced different symptoms in its hosts. In 1959, a freeze-dried vile containing this altered VEE was accidentally dropped by Soviet medical personnel in a stairwell and infected 20 laboratory staff.

     A report in 1998, claimed that the chimeral VEE-smallpox virus did indeed exist and that it produced a smallpox-like illness but with brain symptoms.
     In of 1998, another report claimed that the apartheid government of South Africa had initiated research in order to develop a genetically engineered biological weapon, specifically targeting blacks.
     Theoretically, it is possible to genetically engineer a virus or toxin-synthesizing gene in a bacterium which can be 'activated' by a specific gene or activated by binding to a specific receptor that determines a certain 'ethnic' characteristic (such as skin pigment or eye color). Similar genetic alterations have been achieved in cancer treatment where a unique marker (antigen) on a cancer cell is targeted by an engineered antibody against it, destroying that cancer cell.
Olfactory Neve Fibers at the Cribiform Plate

     Naegleria fowleri is a free-living type of  of protist (animal-like unicellular protozoan) typically found in warm bodies of fresh water (ponds, lakes, rivers, hot springs) but also in soil, near warm-water discharges of industrial plants, and unchlorinated swimming pools. The organism is an amoeba. Amoebae are known to cause gastrointestinal and liver disease and secondarily can affect the nervous system but what is particular about N. fowleri is that it can invade and attack the human nervous system directly. In the natural environment, although nervous system infection with this organism is rare, when it does occur, the fatality rate is close to 100%.
      Human disease caused by this protist was first described in Australia in 1965. Since 1965, more than 144 cases have been confirmed in a variety of countries.

     N. fowleri invades the central nervous system through the nose. The organism penetrates the lining of the nasal passages, resulting in tissue death and hemorrhage. The amoeba then climbs along nerve fibers through the floor of the skull and into the brain. It  then begins to consume the brain cells piecemeal by means of a unique sucking apparatus extended from its cell surface. This results in amoebic meningoencephalitis with a victim survival rate of  less than 1%.
     Delayed diagnoses are a very significant problem to successful treatment of infection, most cases being diagnosed only after death. Infection killed 121 people in the United States from 1937 through 2007, including six in 2007  alone.
     Symptoms of infection usually present about five days after exposure and may include alteration in taste and smell, headache, fever, nausea, vomiting, and stiff neck. The next phase of symptoms consists of confusion, hallucinations, lack of attention, loss of balance, and seizures. The disease progresses rapidly over three to seven days, death occurring from 7- 14 days after exposure.
Route to Infection for
Amoebic Meningoencephalitis

     Of the 133 documented deaths in the US from this infection, 30 people were infected by contaminated recreational water and two people were infected by water from a geothermal (naturally hot) drinking water supply.
     In 2012, in Karachi, Pakistan, the nation's largest city, 10 people died of the 'brain- eating amoeba', allegedly through exposure to contaminated drinking water or bathing water.
     The source of contaminated water was unknown but it was believed that the victims may have picked it up when cleaning out their nostrils - a practice which is common in South Asia. It was suggested that people use boiled or chlorinated water to rinse out their noses and to clean out domestic water tanks which may have become contaminated by the amoeba.
     Vaccines have been developed which protect mice (and presumably humans) from N.fowleri infection. There have been no known instances of the use of this protist infection as a biological weapon. However, there have been numerous articles suggesting that this organism has a huge potential (even if not genetically altered) as an agent which can be used to contaminate water supplies and cause huge damage to enemy military and enemy populations.

     Genetic alteration of N. fowleri, of VEE or of any number of already deadly diseases could easily create an unstoppable epidemic, a disease which targets a particular population but also a disease which can easily 'jump the barrier' (in the same way as animal diseases 'jump the barrier' and affect mankind).
     There is always the risk, however, that the 'chimeral organism' could turn and affect the ones who had taken that step in 'changing the recipe'.

     * Genetic Engineering and Biological Weapons: subject of research for the novel Vaccine - Amazon Kindle.

Biological Warfare in the Twentieth Century

     Early in the 20th century, as microbiology developed, the etiologic agents of many diseases were identified, and many of these were grown and studied in laboratories allowing biological warfare to become more sophisticated.
Horses and Cavalry in WWI

     In the First World War, German and French agents used glanders (also known as EquiniaFarcy, and Malleus),a bacterium (Burkholderia mallei) which occurs in horses and donkeys as well as another animal bacterium called anthrax against enemy forces.
     The German Army also developed cholera to spread in Italy and a wheat fungus specifically for use as biological weapons. They spread plague in St. Petersburg, Russia, infected mules with glanders in Mesopotamia, and attempted to do the same with the horses of the French Cavalry.
Dr. Anton Dilger

     Bioweapons espionage also had its debut in WWI.  A German-American, Dr. Anton Dilger, grew cultures of anthrax as well as glanders bacteria (supplied by the German government), in his Washington, D.C. laboratory. The biological agents were meant to be given to sympathetic dockworkers in Baltimore, Maryland with the goal of infecting some 3000 horses, mules, and cattle being shipped to the Allied troops in Europe.
     Other German secret agents carried out similar campaigns in Romania (infected Romanian sheep were designated for export to Russia) and the US (1915-1916), Argentina (1916-1918) and Spain and Norway. The Germans are also alleged to have dropped biological bombs over Britain.
     On June 17, 1925, the Protocol for the Prohibition of the Use in War of Asphyxiating, Poisonous or Other Gases and of Bacteriological Methods of Warfare (Geneva Protocol of 1925) was signed then registered through the League of Nations.  A total of 108 nations signed the agreement but the Geneva Protocol did not address verification or compliance, making it unenforceable.

     But the Geneva Protocol did not prohibit research or development of these agents and several countries that were parties to the Geneva Protocol of 1925 (Belgium, Canada, France, Great Britain, Italy, the Netherlands, Poland, Japan, and the Soviet Union) began to develop biological weapons even after its ratification.
     The USA did not ratify the Geneva Protocol until 1975.
     In the years which preceded WWII, Japan conducted biological weapons beginning in 1932 and continuing until the end of World War II. The Japanese military practiced biological warfare against China, mostly through the division of the Japanese Imperial Army known as Unit 731 (see post: Death by Physician).
'Work' at Unit 731

     Unit 731 was established in occupied Manchuria  near the town of Pingfan, in 1936 (another, Unit 100 operated out of the city of Changchun) and used human subjects to test the lethality of disease agents such as anthrax, cholera, typhoid, syphilis, meningitis and plague.
     Experiments were also carried out with terodotoxin, an extremely poisonous fungal toxin. In active military campaigns during World War II, several hundred thousand Chinese civilians fell victim to biological attacks by the Japanese.
     In October 1940, the Japanese dropped paper bags filed with plague-infested fleas and grain over the cities of Ningbo and Quzhou in Zhejiang province. The grain attracted rats, which became infected from the fleas, and subsequently spread the disease further into the nearby human population.
     On several occasions, the fleas were released from aircraft over Chinese cities to initiate plague epidemics.
     But, Japanese officials had not adequately prepared, trained, or equipped their own military personnel for the hazards of biological weapons. An attack on the city of Changteh in 1941 reportedly led to approximately 10,000 casualties due to biological weapons - 1700 of them among Japanese troops. The 'field trials' were terminated in 1942.
     During the war, the Japanese Imperial Army used over 3000 American, Korean, British, Australian, Soviet, and Mongolian prisoners of war as guinea pigs.
Gruinard Island

      It was not only the Japanese who looked into the use of biological weapons. In England, scientists experimented with biological agents testing anthrax bombs on Gruinard Island off the northwest coast of Scotland in 1942 and 1943 and then prepared and stockpiled anthrax-laced cattle cakes for the same reason.
     These experiments resulted in severe contamination of the island with persistence of viable spores. In 1986, the island was finally decontaminated by using formaldehyde and seawater. The United States, the army developed Camp Dietrick (now Fort Detrick), Maryland, into a site for biological research and development.
     German medical researchers infected prisoners with disease-producing organisms such as Rickettsia prowazekii, hepatitis A virus, and malaria. Supposedly, Hitler had issued orders prohibiting the development of biological weapons, but all the same, German scientists did establish a biological weapons research.
Dr. Joseph Goebbels

     High-ranking Nazi, Dr. Joseph Goebbels accused the British of attempting to introduce yellow fever into India by importing infected mosquitoes from West Africa.
In 1942, the United States formed the War Research Service under the authority of which anthrax and botulinum toxin were investigated for use as weapons and large quantities of both were stockpiled by June 1944.
     During the Second World War, many countries operated programs focused on the development of biological weapons.
     In the UK, aside from anthrax, research looked at animal and crop diseases and foot and mouth disease (Aphtae epizooticae).
     In Canada, the focus was on anthrax and rinderpest.
     In the Soviet Union, experimenters concentrated their efforts on typhus and plague and in the USA, most research  focused on anthrax and chemical herbicides.
Foot and Mouth Disease


     During the years following World War II, newspapers were filled with articles about disease outbreaks caused by foreign agents armed with biological weapons.
     In the USA, an offensive biological warfare program was started in 1942 under the direction of a civilian agency, the War Reserve Service.
     The program included the research and development facility at Camp Detrick, Maryland (renamed Fort Detrick in 1956) known today as the US Army Medical Research Institute of Infectious Diseases (USAMRIID), testing sites in Mississippi and Utah, and a production facility in Terra Haute, Indiana.
     About 5000 bombs filled with anthrax spores were produced at Camp Detrick. At Fort Detrick, biological munitions were detonated inside a hollow 1-million-liter, metallic, spherical aerosolization chamber known as the eight ball.
     Volunteers inside this chamber were exposed to Francisella tularensis and Coxiella burnetii. The studies were conducted to determine the vulnerability of humans to certain aerosolized pathogens.
     During the Korean War, the Soviet Union, China, and North Korea accused the USA of using agents of biological warfare against North Korea.
     The US program into bioweapons expanded during the Korean War (1950–1953) with the establishment of a new production facility in Pine Bluff, Arkansas and, by the late 1960s, the US military had developed a biological arsenal that included numerous biological pathogens, toxins, and fungal plant pathogens that could be directed against crops to induce crop failure and famine.

The Mau Mau Uprising
     The Mau Mau, protesting British rule in Africa used plant toxins to kill livestock in 1950.
     During the Vietnam War, Viet Cong guerrillas used needle-sharp punji sticks dipped in feces to cause severe infections after an enemy soldier had been stabbed and set up contaminated 'spear traps', impaling unsuspecting victims who fell in.
     In the post-WWII period, there have been several allegations of biological weapon use.
     In 1957, the Eastern European press accused Great Britain of using biological weapons in Oman.
     The Chinese alleged that the USA caused a cholera epidemic in Hong Kong in 1961.
Captured Viet Cong Guerrillas

     In July 1964, the Soviet newspaper Pravda asserted that the US Military Commission in Columbia and Colombian troops had used biological agents against peasants in Colombia and Bolivia.
     In 1969, Egypt pointed a finger at 'imperialistic aggressors' and their use of biological weapons in the Middle East, causing an epidemic of cholera in Iraq in 1966.
     Biological weapons, as they became more sophisticated, grew into an 'industry', funded by many different countries. The increased production of these weapons, however, spawned biological accidents and biological terrorists.
Logo of the Weather Underground
     In 1970, members of the Weather Underground (the Weathermen), a left-wing group in the USA opposed to American imperialism and the Vietnam War, attempted to obtain biological weapons from Fort Detrick, Maryland to contaminate the water supply systems of US cities.
     Members of the right-wing group Order of the Rising Sun were arrested in Chicago in 1972 and had, in their possession 30 to 40 kg of typhoid cultures they had intended to use to poison the water supply in Chicago, St. Louis.
The Aral Sea

     In 1972, police arrested two teenagers who had visions of eliminating humanity so that the group's members could start a new master race. The teens already gathered biological agents, including the typhus bacillus.
     The first example of state-supported bioterrorism in recent history occurred on September 7, 1978. In a James Bond style slaying, Bulgarian exile Georgi Markov, while in London, was injected in the leg with a steel ball impregnated with ricin via a specially constructed umbrella (the 'umbrella killing'). Within 5 hours, Markov became extremely ill and died within 4 days. The communist Bulgarian government had carried out the assassination with technology supplied by the Soviet Union.
   Renaissance Island in the Aral Sea

     Ricin is a highly toxic, naturally occurring protein derived from the seeds of the castor bean plant. A dose as small as a few grains of salt can kill an adult human.

     Ricin is poisonous when inhaled, injected or ingested and acts as a toxin by the inhibition of protein synthesis (type 2 ribosome inactivating protein). Because the symptoms are caused by failure to make protein, they emerge only after a delay of a few hours to a full day after exposure.
     In April 1979, an epidemic of anthrax occurred among the citizens of Sverdlovsk (today known as Ekaterinburg), Russia, killing 66 people.
Emblem of the Order of the Rising Sun

     The epidemic occurred among people who lived and worked near a Soviet military microbiology facility (Compound 19) in Sverdlovsk. In addition, many livestock died of anthrax in the same area, out to a distance of 50 km.
     Investigations revealed that victims died of 'inhalational anthrax', an accidental release of anthrax spores from a biological weapons program, dispersed in an aerosol cloud, downwind from Compound 19.

     After the anthrax incident in Sverdlovsk, the Soviets continued bioweapons research at a remote military facility in the isolated city of Stepnogorsk in Kazakhstan, producing an even more virulent strain of anthrax. In 1980, the former Soviet Union expanded its bioweapons research program and was eventually able to weaponize smallpox  conducted at remote facilities in Siberia.
Georgi Markov

     A primitive laboratory was found in 1980 in Germany in a Red Army Faction (a 1980s terrorist organization) safe house in Paris, reportedly containing a bathtub filled with flasks of Clostridium botulinum.
     In September and October, 1984 in Oregon, members of the Rajneeshee religious cult contaminated restaurant salad bars with Salmonella typhimurium to prevent townspeople from participating in local elections. Over 750 were poisoned and 40 hospitalized.
Castor Bean Plants (Source of Ricin)
     In 1985, Iraq began an offensive biological weapons program producing anthrax, botulinum toxin, and aflatoxin. Aflatoxins are naturally occurring fungal toxins, produced by many species of the fungus, Aspergillus. High-level exposure to aflatoxin produces an acute liver disease and/or liver cancer.
     During Operation Desert Storm (see post: Gulf War Syndrome), allied forces expected the threat of chemical and biological agents. Following the Persian Gulf War, Iraq disclosed that it had indeed fabricated bombs, Scud missiles, 122-mm rockets, and artillery shells armed with botulinum toxin, anthrax, and aflatoxin. They also had spray tanks fitted to aircraft capable of distributing agents over a specific target.
Insignia of the Red Army Faction

     At the end of the First Gulf War in August 1991, representatives of the Iraqi government announced to representatives from the UN Special Commissions Team 7 that Iraq had conducted research into the offensive use of B.anthracis, botulinum toxins, and Clostridium perfringens. Iraq had extensive research facilities at Salman Pak, Al Hakam, and other sites, only some of which were destroyed during the war.
     In the early 1990s, members of the Minnesota Patriots Council , a right-wing extremist militia group, attempted to poison an Internal Revenue Service official, local law enforcement, and a US deputy marshal with ricin.
Aspergillus Fungus

     Bioterrorism resurfaced on March 18, 1995, when the Aum Shinrikyo (see post: Death Cults) attacked the Tokyo subway system with sarin gas. Investigations disclosed evidence of a rudimentary biological weapons program.

     Before March 1995, the cult had attempted unsuccessful biological attacks in Japan using botulinum toxin - April 1990, cult members outfitted a car to disperse botulinum toxin through an exhaust system and drove the car around the Japanese parliament building; March 1995, cult members planted 3 briefcases designed to release botulinum toxin in a Tokyo subway.
Aum Shinrikyo Logo

     In June 1993, Aum Shinrikyo members attempted, over a 4-day period, to spread anthrax in Tokyo via a sprayer system from the roof of a downtown building. Q fever was also acquired by the group - Q fever is a disease caused by infection with Coxiella burnetti, a bacterium which affects humans and other animals. Cult members had also attempted to acquire Ebola virus in Zaire during a trip to that country in 1992.
     In 1996, an Ohio man attempted to obtain bubonic plague cultures through the mail. and in the fall of 2001, anthrax was delivered through the US Postal Service to U.S. media and government offices. The anthrax-contaminated letters resulted in 22 cases of anthrax - 11 confirmed as inhalation anthrax and 11 confirmed as cutaneous with six people dying from the disease.
An Anthrax Letter

     In December 2002, six people were arrested in Manchester, England, their apartment was serving as a 'ricin laboratory'. On Jan. 5, 2003, British police raided two residences around London and found traces of ricin, which led to an investigation of a possible Chechen separatist plan to attack the Russian embassy with the toxin.
     On Feb. 3, 2004, three U.S. Senate office buildings were closed after the toxin ricin was found in a mailroom that served the office of Senate Majority Leader Bill Frist.
     Almost since the beginning of human 'civilization' and especially during the past century, the progress made in biotechnology and biochemistry has simplified the development and production of 'germ' weapons. Indeed, in certain circumstances and regions, biological warfare agents may be more potent than conventional or chemical weapons.
     Biological weapons are unique in that they are invisible and often have delayed effects. These factors allow those who use them to instill fear and cause confusion among their victims and to escape undetected. A germ-warfare attack would cause sickness and death in a large number of victims but would also create fear, panic, and paralyzing uncertainty.
Genetic Engineering?

     Genetic engineering with the potential to develop 'designer' germs holds perhaps the most dangerous potential.
     Today, even 'third world', developing countries often have the desire to possess their own biological 'defences' and ease of production as well as the broad availability of biological agents and technical know how have led to a further spread of these weapons of mass destruction.
     *Biological Weapons in the Twentieth Century: subject of research for the novel Vaccine - Amazon Kindle.