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).
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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.
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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'.
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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.
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Tularemia |
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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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Ameba |
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.
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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'.