An As ssignment on t

GMO Risk for an Os: k nimal o huma heal or an lth Cou Nam GMOs and Bio urse me: s osafety Re egulation n Course No: BTe -408

Subm mitted to, t Anzan Parvi na in Lectu urer, Dept of Biote echnolog & gy Genet Engi tic ineering g, Islam University, K mic Kushtia. Su ubmitte by, ed Ro No: 0 oll 0617023 Re egistratio No: 5 on 532 Session: 20 006-07 ept iotechno ology & De of Bi Ge enetic E Engineer ring, Islamic Un niversity Kush y, htia.

Page 1

1. Introduction
The rationale for accelerated research in biotechnology has several thrusts; two are to understand the living biological system for advancement of science and to find newer applications for commercialization. As early as the 1980s, attention was focused on recombinant deoxyribonucleic acid (DNA) research, in which specific genes were manipulated in a more elegant way to create new entities of living organisms, the so-called genetically modified organisms (GMOs). With the development of recombinant deoxyribonucleic acid (DNA) technology, the metabolic potentials of microorganisms are being explored and harnessed in a variety of new ways. Over the last 30 years, the ability to modify specific genes in organisms has revolutionized numerous fields of the biosciences, including medicine, agriculture, and basic research into life processes. Now genetic engineering offers the advantages over traditional methods of increasing molecular diversity and improving chemical selectivity. In addition, genetic engineering offers sufficient supplies of desired products, cheaper product production, and safe handling of otherwise dangerous agents. However, when the potential for genetically Modified organisms (GMOs) become a part of world development into a reality, much concern was raised over the safety of such a practice. The U.S. Department of Agriculture (USDA) responded in 1990 by formally establishing the Biotechnology Risk Assessment Research Grants Program (BRARGP). Many possible risks surround GMOs and have already started to affect society on personal, regional, national, and worldwide levels.

2. Topics of Concern
Based upon ICGEBs long-standing activities in biosafety, we have identified the main issues derived from the deliberate introduction of GM crops (and their derived products) into the environment or onto the market of concern today. These have been classified as: Risks for animal and human health Risks for the environment Horizontal gene transfer Risks for agriculture General concerns

Here, the general safety issues associated with GMOs that are Risk for animal and human health are described; these include: Toxicity and food quality/safety Allergenicity Pathogen drug resistance (antibiotic resistance). i. Toxicity and food quqlity/safety: Plants have been an important source of food for humans since the very beginning of humankind. Humans have learned that some plants can be eaten safely, but others cannot. It was also learned that some plants could only be eaten after cooking. Knowledge gained on the safety of different plants was passed on from generation to generation. Also, through much of human history, humans have been selecting and breeding comestible plants for desirable properties and traits, such as higher yield or better nutrition. As a result, food plants have been changed dramatically in their morphology, growth, performance, and nutritional composition.

Page 2

With the advances in science and technology, humans have learned more about the chemical composition of plants. It was revealed that food plants are composed of many different compounds and molecules that provide humans with essential macro- and micronutrients and energy. It was also found that, even in some highly domesticated, common food crops, in addition to the various nutrients, there are certain substances that either are toxic or have antinutritional effects to humans and animals; these are called toxicants or antinutrients. Examples of these substances are glycoalkaloids and solanine in potato; erucic acid, phytic acid, and glucosinolates in rapeseed; tomatine in tomato; and trypsin inhibitors in the grains of wheat, rice, and corn. Safety for consumption for some plant foods is relative because safety margins for many natural plant toxicants are quite small For example, a vegetable, Sauropus androgynus, which was safely eaten in Borneo and Malaysia, caused severe toxicity and death when consumption increased because of a new perception that it was a health food. Besides small compounds, plants may also produce protein toxins. As an example, a family of proteins called -thionins is a group of small (5-kDa), highly basic, disulfide rich proteins found in seeds, stems, roots, and leaves of several common food plants, such as corn and barley. The different members of this family of plant proteins show both sequence and structural homology and are toxic to bacteria, fungi, yeasts, and animal cells. In fact, there are structural similarities between -thionins of plants and -conotoxins, a group of toxins isolated from the venom of the piscivorous sea snail Conus geographus. It was reported that plant -thionins and sea snail conotoxins share a similar mode of action by blocking sodium channels of the cells. a. Nutritional/toxicological studies: Herbicideresistant soybean: Studies have been conducted on the feeding value and possible toxicity for rats, broiler chickens, catfish and dairy cows of two GM lines of glyphosate-resistant soybean (GTS). The growth, feed conversion efficiency, catfish fillet composition, broiler breast muscle and fat pad weights and milk production, rumen fermentation and digestibilities in cows were claimed to be similar for GTS and non-GTS. However: These experiments were poorly designed since the high dietary protein concentration and the low inclusion level of GTS could have masked any GM effect. No individual feed intakes, body or organ weights were given and no histology was performed, except some qualitative microscopy on the pancreas. The feeding value of the two GTS lines was not substantially equivalent either because the rats grew significantly better on one of the GTS lines than on the other. The experiment with broiler chicken was a commercial and not a scientific study. The catfish experiment showed again that the feeding value of one of the GTS lines was superior to the other. Milk production and performance of lactating cows also showed significant differences between cows fed GM and non-GM feeds. Moreover, testing of the safety of 5-enolpyruvylshikimate-3-phosphate synthase which renders soybeans glyphosate-resistant was irrelevant because in the gavage studies an E. coli recombinant and not the GTS product was used. Their effects could be different as the differences in post-translational modification could have impaired their stability to gut proteolysis.

Thus, the claim that the feeding value of GTS and non-GTS lines was substantially equivalent is at best premature. In a separate study it was claimed that rats and mice which were fed 30% toasted GTS or nonGTS in their diet had no significant differences in nutritional performance, organ weights, histopathology and production of IgE and IgG antibodies. However, under the unphysiological basically, starvation conditions of these experiments when, instead of the normal daily growth of 5-8 g per day, the rats grew less than 0.3 g and mice not at all, no valid conclusions could be drawn.

Page 3

Result: Rats had meager weight gain when fed GM soybeans. GM corn: One broiler chicken feeding study with rations containing transgenic Event 176 derived Bt corn (Novartis) has been published. However, the results of this trial are more relevant to commercial than academic scientific studies. GM peas: The nutritional value of diets containing GM peas expressing bean alpha-amylase inhibitor when fed to rats for 10 days at two different (30% or 65%) dietary inclusions was shown to be similar to that of parent-line peas. Even at 65% level the difference was small mainly because the alpha-amylase inhibitor expressed in the peas was quickly digested in the rat gut and its antinutritive effect abolished. Unfortunately no gut histology was done or lymphocyte responsiveness measured. Although some organ weights, mainly the caecum and pancreas were different, those of others were remarkably similar suggesting that GM peas may be used in the diets of farm animals at low/moderate levels if their progress was carefully monitored.

However, to establish its safety for humans a more rigorous specific risk assessment will have to be carried out with several GM lines. This should include: An initial nutritional/toxicological testing on laboratory animals If no harmful effects are then detected, it should be followed by clinical, double-blind, placebo-type tests with human volunteers, keeping in mind that any possible harmful effects would be particularly serious with the young, old, and disabled.

A protocol for such testing was given at the OECD conference in Edinburgh, February 2000 and subsequently published. Result: GM peas seem to have no harmful effects on animals but that doesnt mean they are safe for humans. GM potatoes: In a short feeding study to establish the safety of GM potatoes expressing the soybean glycinin gene, rats were daily force-fed with 2 g of GM or control potatoes/kg body weight. Although no difference in growth, feed intake, blood cell count and composition and organ weights between the groups was found, the potato intake of the animals was too low and unclear, whether the potatoes were raw or boiled. Feeding mice with potatoes transformed with a Bacillus thuringiensis var. kurstakiCry1 toxin gene or the toxin itself was shown24 to have caused villus epithelial cell hypertrophy and multinucleation, disrupted microvilli, mitochondrial degeneration, increased numbers of lysosomes and autophagic vacuoles and activation of crypt Paneth cells. The results showed that despite claims to the contrary, CryI toxin was stable in the mouse gut and therefore GM crops expressing it need to be subjected to thorough teststo avoid the risks before marketing. In another study, young, growing rats were pair-fed on iso-proteinic and iso-caloric balanced diets containing raw or boiled non-GM potatoes and GM potatoes with the snowdrop (Galanthus nivalis) bulb lectin (GNA) gene. The results showed that the mucosal thickness of the stomach and the crypt length of the intestines of rats fed GM potatoes was significantly increased. Most of these effects were due to the insertion of the construct and not to GNA which had been been preselected as a non-mitotic lectin unable to induce hyperplastic intestinal growth and epithelial T lymphocyte infiltration. Although there is controversy about the tests, most of the adverse comments on this Lancet paper were personal, non-peer reviewed opinions and, as such, of limited scientific value. The findings, on the other hand, were published in a peer-reviewed publication25 and the criticism replied to. The work, however, has not been repeated nor results contradicted and it is therefore imperative that the effects on the gut structure and metabolism of all other GM crops developed using similar techniques and genetic vectors should be thoroughly investigated before their release into the food chain.

Page 4

Result: Toxins were found in mice after eating GM potatoes. GM tomatoes: This study with a GM tomato expressing B. thuringiensis toxin CRYIA(b) gene was published in a book and not in a peer-reviewed journal. However, its importance was underlined by the immunocytochemical demonstration of in vitro binding of Bt toxin to the caecum/colon from humans and rhesus monkeys. Although in vivo the Bt toxin was not bound by the rat gut, this was possibly due to the authors use of recombinant Bt toxin. b. Compositional studies: GM soybeans: To make soybeans herbicide resistant, the gene of 5-enolpyruvylshikimate-3phosphate synthase from Agrobacterium was used. Safety tests claim the GM variety to be substantially equivalent to conventional soybeans. The same was claimed for GTS (glyphosateresistant soybeans) sprayed with this herbicide. However, several significant differences between the GM and control lines were recorded and the statistical method used was flawed because: Instead of comparing the amounts of components in a large number of samples of each individual GTS with its appropriate parent line grown side-by-side and harvested at the same time, the authors compared samples from different locations and harvest times. There were also differences in the contents of natural isoflavones (genistein, etc.) with potential importance for health. Additionally, the trypsin inhibitor (a major allergen) content was significantly increased in GTS. Because of this, and the large variability ( 10% or more), the lines could not be regarded as substantially equivalent. Result: Allergen content increased when soybeans were genetically modified. GM potatoes: There is only one peer-reviewed publication on GM potatoes that express the soybean glycinin gene. However, the expression level was very low and no improvements in the protein content or amino acid profile were obtained. GM rice: The kind that expresses soybean glycinin gene (40-50 mg glycinin/g protein) has been developed and is claimed to contain 20% more protein. However, the increased protein content was probably due to a decrease in moisture rather than true increase in protein putting a question mark over the significance of this GM crop. GM cotton: Several lines of GM cotton plants have been developed using a gene from Bacillus thuringiensis subsp. kurstaki providing increased protection against major lepidopteran pests. The lines were claimed to be substantially equivalent to parent lines in levels of macronutrients and gossypol, cyclopropenoid fatty acids and aflatoxin levels were less than those in conventional seeds. However, because of the use of inappropriate statistics it is questionable whether the GM and non-GM lines were truly equivalent, particularly as environmental stresses could have unpredictable effects on antinutrient/toxin levels. Result: The toxin level of GM cotton is unpredictable. c. Safety tests on commercial GM crops: GM tomatoes: The first and only safety evaluation of a GM crop, the FLAVR SAVRTM tomato, was commissioned by Calgene, as required by the FDA. This GM tomato was produced by inserting kanr genes into a tomato by an antisense GM method. The test has not been peerreviewed or published but is on the internet. The results claim there were no significant alterations in total protein, vitamins and mineral contents and in toxic glycoalkaloids. Therefore, the GM and parent tomatoes were deemed to be substantially equivalent.

Page 5

In acute toxicity studies with male/female rats, which were tube-fed homogenized GM tomatoes, toxic effects were claimed to be absent. In addition, it was concluded that mean body and organ weights, weight gains, food consumption and clinical chemistry or blood parameters were not significantly different between GM-fed and control groups. However: The unacceptably wide range of rat starting weights (18% to 23%) invalidated these findings. No histology on the intestines was done even though stomach sections showed mild/moderate erosive/necrotic lesions in up to seven out of twenty female rats but none in the controls. However, these were considered to be of no importance, although in humans they could lead to life-endangering hemorrhage, particularly in the elderly who use aspirin to prevent thrombosis. Seven out of forty rats on GM tomatoes died within two weeks for unstated reasons. These studies were poorly designed and therefore the conclusion that FLAVR SAVRTM tomatoes were safe does not rest on good science, questioning the validity of the FDAs decision that no toxicological testing of other GM foods will in future be required.

Result: Some rats died within a few weeks after eating GM tomatoes. GM maize: Two lines of Chardon LL herbicide-resistant GM maize expressing the gene of Phosphinothricin Acetyltransferase Enzyme (PAT-PROTEIN) before and after ensiling showed significant differences in fat and carbohydrate contents compared with non-GM maize and were therefore substantially different. Toxicity tests were only performed with the PAT-PROTEIN even though with this the unpredictable effects of the gene transfer or the vector or gene insertion could not be demonstrated or excluded. The design of these experiments was also flawed because: The starting weight of the rats varied by more than 20% and individual feed intakes were not monitored. Feed conversion efficiency on PAT-PROTEIN was significantly reduced. Urine output increased and several clinical parameters were also different. The weight and histology of the digestive tract (and pancreas) was not measured.

Thus, GM maize expressing PAT-PROTEIN may present unacceptable health risks. Result: Rats ability to digest was decreased after eating GM corn. ii. Allergenicity: Allergies are caused by the specific activation of an inflammatory process resulting from allergens interacting with immune effector mechanisms. Specific allergies show geographic distribution primarily because of dietary considerations. Some of the best-documented examples of food allergies include peanuts, milk, hens eggs, Brazil nuts, hazelnuts, walnuts, shellfish, celery, kiwi fruit, and rice. Although typical symptoms of food allergens are not life threatening, severe anaphylactic reactions may be fatal. The term food allergy is often overused by the public, as well as by some physicians and scientists, to describe any undesired or bothersome problem related to diet. For clarity, human adverse reactions to foods can be classified into two categories: toxic reactions and nontoxic reactions. Toxic reactions are caused by toxicants naturally occurring in foods or toxic compounds and contaminants introduced during food production, processing, and handling. Nontoxic reactions are further divided into food intolerance and food allergy. Food intolerance refers to those nontoxic reactions that are nonimmune mediated. One example of food intolerance is lactose intolerance, the inability to digest lactose present in dairy food products. Food allergy is defined as those nontoxic reactions that are primarily immune mediated.

Page 6

The antigenic molecules giving rise to the immune response are therefore called food allergens. The immune-mediated food allergy can be mediated by IgE (immunoglobulin E) or non-IgE mediated. Most known cases of food allergy are IgE mediated. In IgE-mediated food allergy, when first ingested, the allergens may be cleaved to some degree by digestive enzymes, absorbed by the gut mucosa, processed in immunopotent cells, and then presented to the immune system, resulting in the production of allergen-specific IgE. The allergen-specific IgE antibodies circulate in the body and will bind to mast cells throughout the body tissues and basophils in the blood. On renewed contact with the food allergens, the allergens bind to the IgE antibodies, which triggers the degranulation of the mast cells and basophils and production of mediators (such as histamine and leukotrienes), which induce various clinical symptoms. Symptoms of IgE-mediated food allergy vary among individuals and are nonspecific. The clinical aspects of IgE-mediated food allergy can be oral (itching and swelling of lips, mouth, or throat); dermal (urticaria, atopic dermatitis, angioederma); gastrointestinal (vomiting, cramps, diarrhea, abdominal pain); respiratory (asthma, rhinitis, bronchospasm, wheezing); and cardiovascular (decrease in blood pressure, anaphylaxis). The symptoms may occur within minutes to days after ingestion of the offending food, and in severe cases, death could occur. Allergenicity studies: One of the major health concerns with GM food is its potential to increase allergies and anaphylaxis in humans eating unlabeled GM foodstuffs. When the gene is from a crop of known allergenicity, it is easy to establish whether the GM food is allergenic usingin vitrotests, such as RAST or immunoblotting, with sera from individuals sensitised to the original crop. This was demonstrated in GM soybeans expressing the brasil nut 2 S protein or in GM potatoes expressing cod protein genes. It is also relatively easy to assess whether genetic engineering affected the potency of endogenous allergens. Some farm workers exposed to B. thuringiensis pesticide were shown to have developed skin sensitization and IgE antibodies to the Bt spore extract. With their sera it may now therefore be possible to test for the allergenic potential of GM crops expressing Bt toxin. It is all the more important because Bt toxin Cry1Ac has recently been shown to be a potent oral/nasal antigen and adjuvant.

Assessment of the allergenicity of a GM foodcrop, however, is difficult when the gene is transferred from a source not eaten before or with unknown allergenicity or on gene transfer/insertion a new allergen or adjuvant is developed or the expression of a minor allergen is increased. Unfortunately, while there are good animal models for nutritional/toxicological testing, no such models exist for allergenicity testing. Presently only indirect and rather scientifically unsound methods, such as finding SHORT sequence homologies (at least 8 contiguous amino acids) to any of the about 200 known allergens, are used for the assessment of allergenicity. The decision-tree type of indirect approach based on factors (such as size and stability) of the transgenically expressed protein is even more unsound, particularly as its stability to gut proteolysis is assessed by an in vitro (simulated) testing instead of in vivo(human/animal) testing and this is fundamentally wrong. The concept that most allergens are abundant proteins is also misleading because for example Gad c 1, the major allergen in codfish, is not a predominant protein. However, when the gene responsible for the allergenicity is known, such as the gene of the alpha-amylase/trypsin inhibitors/allergens in rice, cloning and sequencing opens the way for reducing their level by antisense RNA strategy.

Thus, in the absence of reliable methods for allergenicity testing, it is at present impossible to definitely establish whether a new GM crop is allergenic or not before its release into the human/animal food/feed chain.

Page 7

iii. Pathogen drug resistance (Antibiotic resistance): The combination of antibiotic resistance genes and antibiotic is an important tool in genetic engineering in general and in plant biotechnology in particular. A key task in genetic engineering is the identification and selection of cells into which a new gene has been introduced. Antibiotic resistance genes have the ability to selectively inactivate certain antibiotics and consequently protect cells against these antibiotics. An antibiotic resistance gene can thus be used to tag a gene carrying a trait or characteristic of interest. In practice the antibiotic gene is linked to a gene carrying the trait of interest prior to its introduction into the recipient cells, whether of bacterial, yeast, plant or animal origin. These cells are then incubated in the presence of the antibiotic. The only cells which multiply under these conditions are the ones that have incorporated the antibiotic resistance gene together with the trait of interest. However, concern has been expressed that the release of these markers in GM plants may result in an increase in the rate of antibiotic resistance in human pathogens. a. Bacterial mechanisms of antibiotic resistance: Several mechanisms have evolved in bacteria which confer them with antibiotic resistance. These mechanisms can chemically modify the antibiotic, render it inactive through physical removal from the cell, or modify target site so that it is not recognized by the antibiotic. The most common mode is enzymatic inactivation of the antibiotic. An existing cellular enzyme is modified to react with the antibiotic in such a way that it no longer affects the microorganism. An alternative strategy utilized by many bacteria is the alteration of the antibiotic target site. These and other mechanisms are shown in the the figure and accompanying table below.

Table: Mechanism of antibiotic resistance Antibiotic Method of resistance Chloramphenicol Reduced uptake into cell Tetracycline Active efflux from the cell -lactams, Erythromycin, Eliminates or reduces binding of antibiotic to cell target Lincomycin -lactams, Aminoglycosides, Enzymatic cleavage or modification to inactivate Chloramphenicol antibiotic molecule Sulfonamides, Trimethoprim Metabolic bypass of inhibited reaction Sulfonamides, Trimethoprim Overproduction of antibiotic target (titration)

Figure: Mechanisms of antibiotic resistance in bacteria

Page 8

b. The acquisition and spread of antibiotic resistance in bacteria: The development of resistance is inevitable following the introduction of a new antibiotic. Initial rates of resistance to new drugs are normally on the order of 1%. However, modern uses of antibiotics have caused a huge increase in the number of resistant bacteria. In fact, within 8-12 years after wide-spread use, strains resistant to multiple drugs become widespread. Multiple drug resistant strains of some bacteria have reached the proportion that virtually no antibiotics are available for treatment. Antibiotic resistance in bacteria may be an inherent trait of the organism (e.g. a particular type of cell wall structure) that renders it naturally resistant, or it may be acquired by means of mutation in its own DNA or acquisition of resistance-conferring DNA from another source. Inherent (natural) resistance: Bacteria may be inherently resistant to an antibiotic. For example, an organism lacks a transport system for an antibiotic; or an organism lacks the target of the antibiotic molecule; or, as in the case of Gram-negative bacteria, the cell wall is covered with an outer membrane that establishes a permeability barrier against the antibiotic. Acquired resistance: Several mechanisms are developed by bacteria in order to acquire resistance to antibiotics. All require either the modification of existing genetic material or the acquisition of new genetic material from another source. Vertical gene transfer: The spontaneous mutation frequency for antibiotic resistance is on the order of about of about 10-8- 10-9. This means that one in every every 108- 109 bacteria in an infection will develop resistance through the process of mutation. In E. coli, it has been estimated that streptomycin resistance is acquired at a rate of approximately 10-9 when exposed to high concentrations of streptomycin. Although mutation is a very rare event, the very fast growth rate of bacteria and the absolute number of cells attained means that it doesn't take long before resistance is developed in a population. Once the resistance genes have developed, they are transferred directly to all the bacteria's progeny during DNA replication. This is known as vertical gene transfer or vertical evolution. The process is strictly a matter of Darwinian evolution driven by principles of natural selection: a spontaneous mutation in the bacterial chromosome imparts resistance to a member of the bacterial population. In the selective environment of the antibiotic, the wild type (non mutants) are killed and the resistant mutant is allowed to grow and flourish Horizontal gene transfer: Another mechanism beyond spontaneous mutation is responsible for the acquisition of antibiotic resistance. Lateral or horizontal gene transfer (HGT) is a process whereby genetic material contained in small packets of DNA can be transferred between individual bacteria of the same species or even between different species. There are at least three possible mechanisms of HGT, equivalent to the three processes of genetic exchange in bacteria. These are transduction, transformation or conjugation. Conjugation occurs when there is direct cell-cell contact between two bacteria (which need not be closely related) and transfer of small pieces of DNA called plasmids takes place. This is thought to be the main mechanism of HGT. Transformation is a process where parts of DNA are taken up by the bacteria from the external environment. This DNA is normally present in the external environment due to the death and lysis of another bacterium. Transduction occurs when bacteria-specific viruses (bacteriophages) transfer DNA between two closely related bacteria.

Page 9

c. Societal, medical and agricultural practices that lead to antibiotic resistance: In the face of a microbe's inherent ability to develop antibiotic resistance, many societal. medical and agricultural practices contribute to this process, foremost of which are discussed below. Antibiotics in food and water: Prescription drugs are not the only source of antibiotics in the environment. In the United States, antibiotics can be found in beef cattle, pigs and poultry. The same antibiotics then find their way into municipal water systems when the runoff from housing facilities and feedlots contaminates streams and groundwater. So it's a double hit: we get antibiotics in our food and drinking water, and we meanwhile promote bacterial resistance. Routine feeding of antibiotics to animals is banned in the European Union and many other industrialized countries. Maybe they know something we don't. Indiscriminateuseofantibioticsinagricultureandveterinarypractice:The non-therapeutic use of antibiotics in livestock production makes up at least 60 percent of the total antimicrobial production in the United States. Irresponsible use of antibiotics in farm animals can lead to the development of resistance in bacteria associated with the animal or with people who eat the animal. Such resistance can then be passed on to human pathogens by mechanisms of HGT. Of major concern is the use of antibiotics as feed additives given to farm animals to promote animal growth and to prevent infections (rather than cure infections). The use of an antibiotic in this way contributes to the emergence of antibiotic-resistant pathogens and reduces the effectiveness of the antibiotic to combat human infections. Antibiotic resistance in genetically modified crops: Antibiotic-resistance genes are used as "markers" in genetically modified crops. The genes are inserted into the plant in early stages of development to in order to detect specific genes of interest. e.g. herbicide-resistant genes or insecticidal toxin genes. The antibiotic-resistance genes have no further role to play, but they are not removed from the final product. This practice has met with criticism because of the potential that the antibiotic-resistance genes could be acquired by microbes in the environment. In some cases these marker genes confer resistance to front-line antibiotics such as the beta-lactams and aminoglycosides. Inappropriate use of antibiotics in the medical environment: One problem is the casual use of antibiotics in medical situations where they are of no value. This is the fault of both health care workers and patients. Prescribers sometimes thoughtlessly prescribe 'informed' demanding patients with antibiotics. This leads to use of antibiotics in circumstances where they are of not needed, e.g. viral upper respiratory infections such as cold and flu, except when there is serious threat of secondary bacterial infection. Another problem is patient failure to adhere to regimens for prescribed antibiotics. d. Safety assessment of antibiotic resistance in GM crops: The safety assessment of crops containing antibiotic resistance markers has been thoroughly reviewed by experts from internationally recognised scientific bodies and scientific committees in the European Union and governmental experts from Canada, Japan, Switzerland and the United States among other countries. The experts evaluated the safety of crops containing the nptII marker gene conferring resistance to the antibiotics kanamycin and neomycin, the aad gene conferring resistance to streptomycin and spectinomycin and the bla gene conferring resistance to ampicillin. They concluded that the risk of gene transfer from transgenic crops containing these genes to the microbial population is negligible and, if it were to occur, there would be no impact on the present high frequency of occurrence of antibiotic resistance in microbial populations. The European Commission Scientific Committee on Plants provided an opinion on the safety of a Bt insectprotected maize product containing the nptII gene.

Page 10

Their conclusions are based on the following observations: The antibiotic resistance markers used in plant biotechnology were obtained from naturally occurring bacteria. The nptII gene in particular was isolated from bacteria in the human gut. The markers in approved products are already widespread because effective natural transfer mechanisms exist to shuttle the genes directly between bacterial cells. This process confers advantages to bacteria. Some microbial populations can hence act as reservoirs for certain antibiotic resistance genes that then are rapidly spread in response to selective pressures. The antibiotics inactivated by nptII, kanamycin and neomycin are rarely used in human therapy since they have been replaced by less toxic antibiotics with greater efficacy. There is no known mechanism for the transfer of genes from plant cells to bacteria. Gene transfer to soil micro-organisms could in theory occur in fields where GM crops are cultivated. The rate of DNA transfer between bacteria under optimal conditions in nature is in the range of 10-2 to 10-5. The estimated frequency of uptake of the nptII gene marker from transgenic plants to bacteria under optimal conditions is 10-17 and thus considered to be insignificantly small. Antibiotic resistance markers only confer resistance against specific antibiotics. Antibiotic resistance markers do not result in antibiotic production. There are therefore no antibiotics present in food produced from plants produced using biotechnology.

However, alternative markers and marker removal systems are being investigated in response to public concerns and to expand the number of tools available in plant molecular biology.

3. Conclusion
Over 1000 GMOs and their products have been tested in field trials or industrial applications, ultimately finding themselves in natural surroundings. Considering the increasing number of international GMO releases derived from transgenics, it is important not to ignore the issues related to risk. So, for this reason, the progress of GMO use and technology has always been, and still is, accompanied by interesting and controversial discussions. The agricultural biotechnology debate has addressed the practice of technology and containment, that is, how to prevent the accidental release of GMOs into the environment.

References: Sarad Parekh R.: The GMO Handbook Genetically Modified Animals, Microbes, and Plants in Biotechnology. Antibiotic Resistance Markers in Genetically Modified (GM) Crops (Publication). http://textbookofbacteriology.net/themicrobialworld/homepage.html http://www.icgeb.org/~bsafesrv/index.html http://www.actionbioscience.org