GMOs by kimt7410

Understanding

GMOs GENETICALLY MODIFIED ORGANISMS By: Kim Tan

16/12/14

CONTENTS 5

WHAT ARE GMOS ♦ Advantages & Disadvantages

GMOS IN CANADA ♦ Maize, Rapeseed, Soybeans, Sugar beet, Cotton, Papaya, Squash, & Milk Products with rBST

THE PROCESS ♦ Isolation, Cloning, & Induction

GM CROPS ♦ Bt Corn ♦ The Flavr Savr Tomato

GM & ANIMALS ♦ Cows & rBST ♦ Growth Hormone and Antibiotics ♦ GM Fish

22 24

THE HISTORY CONCERNS ♦ Dr. Arpad Pusztai’s Research ♦ Antibiotic Resistance ♦ Bovine Growth Hormone & Cancer

REGULATIONS ♦ Novel Foods in Canada ♦ Food Labelling

CURRENT & PAST ISSUES 2

19 10 22 21

20 15

―Essentially, a GM food is one derived from an organism that has had some of its heritable traits changed. This can involve: traditional breeding techniques of crossbreeding, using chemicals or radiation to alter the genetic make-up of the organism's cells in a process called mutagenesis, or applying recombinant DNA or genetic engineering techniques - for instance, introducing a gene from one species into another species.‖ -Health Canada ―GMO's — or genetically modified organisms — refer to the plants or animals created through the gene splicing techniques of biotechnology. In conversation, GMO's and GE foods refer to the same thing. They are foods created by merging DNA from different species.‖ -David Suzuki Foundation ―Genetically modified crops – also known as genetically modified organisms (GMOs), GE crops or biotech crops – include one or more genes from another organism, such as a bacterium or other microbe or other plant species. For plants, the inserted gene results in a beneficial characteristic in the plant, such as the ability to tolerate environmental pressures from damaging insects or drought. GMO is commonly used to refer to GM plants, as well as the food or ingredients from GM plants.‖ -Monsanto ―Since 1996, Greenpeace is standing up against the spread of genetically engineered organisms also known as genetically modified organisms (GMO). Genetic engineering is the manipulation of genes to create new plants, animals and microorganisms. Multinational corporations have GMO crops planted on millions of hectares of land — a giant genetic experiment with unpredictable and possibly irreversible risks.‖ -Greenpeace

What are

GMOs? G enetically modified organisms (GMOs) are plants or animals with altered genetic material as a result of genetic engineering (GE). Genetic engineering is the process of transferring useful characteristics into plants, animals, or microorganisms by inserting genes from another organism. This is done using processes and tools of biotechnology which is the use of living organisms or substances from living organisms to develop an agricultural, medicinal, or environmental product or process. The main tools used in biotechnology include restriction enzymes, gel electrophoresis, and plasmid vectors. Other organisms that have been genetically modified include bacteria, yeast, and insects.

The common goal of farmers is to improve wild plants and animals. For over 10,000 years, farmers have been doing so by the selective breeding of desired characteristics. The only difference with GMOs is that they are created by means of biotechnology. Desired genes are selected, recombined, and introduced into the existing organism. All crops improved with recombinant DNA have been developed to aid farmers increase productivity by reducing crop damage from weeds, insects, and disease. The first food product of biotechnology appeared on the market in 1994 and by 2003, over seven million farmers in 18 countries were growing GM crops. The six top countries in the GMO industry in order are the US, Argentina, Canada, Brazil,

China, and South Africa. The top 4 GM crops are herbicide-tolerant soybeans, insect-resistant maize, herbicide resistant canola, and Bt cotton. The Flavr Savr Tomato was the first GMO crop to be approved by the FDA in 1994. Since then, over 81 GM foods have been approved in Canada. Common GM crops in Canada include corn, soya, sugar beets, and canola while other GM products such as cottonseed oil, papaya, squash, and milk products are imported. In 20 years, GMOs have become present in almost all the processed foods sold in our grocery stores. Apples, potatoes, and wheat are more foods waiting to be approved. There is a huge controversy over GMOs. Some people argue that GMOs

are not safe to consume while others defend them as being highly beneficial. However, there are both advantages and disadvantages concerning the use of GMOs. ADVANTAGES An advantage to GM foods are the health benefits. Crops can be modified to have a higher nutritional content or to be lower in calories and fats to benefit our health. In developing countries, this can have a huge impact as the people rely heavily on a single stable. For example, rice is the main diet of many people in poorer counties, however it does not have a balanced nutritional content. To counter this, golden rice has been created which is GM rice that stimulates the body to produce vitamin A. This will help the two million children that die a year and 500,000 that go blind due to a vitamin A deficiency. GM crops are also produced with less pesticides than traditional crops, making them more safe to eat. Some foods are even being invented with vaccines in them to treat people for diseases as they consume the product. GMOs designed to be pestresistant and herbicide-tolerant will decrease the amount of crops lost each year due to pests and chemicals. The cost of pesticides and labour will also

be decreased for farmers, resulting in lower food prices for everyone. Even the environment benefits with less pesticides contaminating the crop, soil, wildlife, and groundwater. In 1999, the Environmental Protection Agency (EPA) reported that farmers growing GM cotton had a 21% decrease in insecticide use. Although cross contamination can be a problem for organic farmers, solutions of creating male-sterile crops that do not produce pollen or pollen that does not contain the introduced gene are being studied. Crops designed to be tolerant to a broad spectrum herbicide also allows farmers to leave plants alone after the season instead of plowing the field reducing soil erosion by 70%. Soil erosion is a worldwide problem as well as a loss for farmers as they lose 25 billion tons of topsoil worldwide each year due to weather conditions. A higher yield in crops will prove important in feeding our overpopulated earth. New genes in crops will also allow them to grow in difficult conditions. Scientists are looking to create plants that will survive frosts by injecting them with an antifreeze gene. Drought and salt tolerant crops are also being researched. One GE seed company, Monsanto, reported a 5-8% increase in yield of their GM crops while there has only been a 1-2%

increase in conventional crops. With the raising problem of overpopulation and global warming, these modified crops will prove effective and essential in feeding the world. Some scientists also believe that GMOs are natural. Plants and animals genetically modify themselves all the time through evolution. As well, most of our common foods today are the product of our modification for a better yield, taste, and size. For example, we have created countless vegetables from a species of mustard including broccoli, Brussels sprouts, and cabbage. New processes of genetic engineering are only more precise allowing the modification of limited genes instead of combining thousands of genes at a time. GM companies also conduct in-depth research of their products before they are approved. The effect of the introduced gene on the organismâ&#x20AC;&#x2122;s chemical makeup and nutritional content is analyzed along with its possibility of toxicity and inducing allergies. DISADVANTAGES However, there are many disadvantages of GMOs including the possibility of creating new allergens especially when genes from an unrelated organism is introduced into a

crop. Bacteria in our digestive system may also inherit the antibiotic resistant genes used in the production of GMOs causing them to become increasingly resistant to antibiotics. In addition, the real effects of GMOs in the future cannot be foreseen or known by just a few tests on lab animals. With the patenting of GM seeds and their complementary pesticides, small farmers are being outcompeted by larger farms. Some farmers are also being sued by Monsanto for growing their patented crops without paying when most of the time, they were not at fault and the cause was cross contamination of GM pollen. Poor countries are also outcompeted by richer countries that can afford these new products. Many small farmers lose their land and belongings because although they achieved higher yields, the cost of the seeds were so high they resulted in a loss. GM crops also raise our reliance on a limited number of crops. Only 15 crops make up the 90% of the crops consumed worldwide today. Monoculture increases our vulnerability to pests and diseases which is a high risk with the modern evolution of pests. Furthermore, GM crops pose harm to the environment. Other wildlife may be harmed such as the monarch butterfly that is effected by the pollen

produced by BT corn. There are fears of "superweeds" from weeds crossing with a plant with a herbicide-resistant gene as well as herbicide-resistant crops themselves invading other fields. Viruses may also inherit crop virus genes from GM crops and infect a whole range of plants. Some people also feel that GMOs go against nature and that the genetic makeup of organisms should not be manipulated. The natural change in genetic makeup of organisms take millions of years while genetic engineering allows it to happen in days. In addition, taking the gene of an animal and inserting it into a crop creates a problem for vegetarians, especially in countries where GMO labelling is not mandatory. Biotechnology companies are profit driven and with the amount of money spent on research and development, they will be more concerned with their patent and the profit made from their products rather than health, economical, and environmental issues. The use of GMOs has become a global market with farmers having to purchase products from seed producers. These companies develop and patent GMO seed along with the pesticides that these seeds are resistant to. The largest GMO seed company in the

world is Monsanto owning nearly 86% of GMO seeds grown globally. These companies are also only concerned with crops that are produced in vast quantities such as soybeans, corn, and cotton rather than crops that will feed developing countries in the future. In many countries, GM crops are being rushed to be approved without adequate testing or public input. Labelling of GMOs are also not mandatory in many countries including the Canada and the US. This limits consumers from having the right to choose their food and the ability to trace any illnesses back to GM foods. In conclusion, GMO foods have not been proven to be safe but have been connected to some health and environmental issues. Most nations require mandatory labelling of GMO foods while some have banned GMO food production and imports altogether. In Canada, GMOs are present in almost all of our food products without us knowing as labelling is not mandatory. Most Canadians are unaware of the GMO ingredients in the foods they choose which should be a right. Nevertheless, GMOs come with both positive and negative effects alike many other things. â&#x2122;Ś

Companies using GM ingredients

Starting in 1994, over 81 GM foods have been approved in Canada. There are 4 GM crops being grown in Canada: corn, canola, soy, and sugar beets. These crops are usually processed to be used as ingredients in food products, as biofuel, or animal feed. Canada also imports GM products including papaya, squash, cottonseed oil, and milk (containing Bovine Growth Hormone).

Maize/Corn Maize plays an important role in feeding livestock and the starch industry to be used in numerous foods. In 1996, GM maize was grown in Canada and the US for the first time. It is the only GM crop grown commercially in the EU. GM maize include traits to be insect resistant and herbicide tolerant. Bt corn is insect resistant to a common pest called the European corn borer that chews the stalks causing the plant to fall over. This damage can also lead to fungal diseases and toxic substances such as mycotoxins. The use of Bt and other varieties of maize allow an increase in yield and decrease in cost and use of pesticides. WHERE ON SHELVES: corn cereals, corn chips, popcorn, cornstarch, corn oil, corn ingredients in processed foods such as glucose and fructose

Rapeseed/Canola Rapeseed did not use to be a popular crop until two substances were removed from them. Euric acid, making the rapeseed bitter prevented the use of its oil in food, and gluconsinolates, found in the meal leftover from pressing being toxic prevented its use in animal feed. When these two substances were removed, rapeseed became known as "double-zero" rapeseed, or "canola" in Canada, to differentiate it from other varieties of rapeseed. Today, canola has many applications including cooking oil, margarine production, animal feed, raw material for biodiesel, industrial oils, and lubricants. Canola is genetically modified to be herbicide tolerant to varying herbicides, allowing the use of the specific chemicals without killing the crop. Since 1996, GM canola has been grown in Canada. In 2007, 87% (5.1 million hectares) of the canola grown in Canada was GM. Canola is currently being developed to have a higher content of long fatty acid chains allowing a higher melting point and margarine that can be produced from few processing steps. Another modification being made is to have a higher content of mid-sized fatty acid chains by including a gene for the production of lauric acid. Lauric acid, usually found in coconut oil to be used as a detergent additive, will enable canola to be used as fat coatings in food processing. WHERE ON SHELVES: canola oil, eggs, milk, and meat

Soybeans Soybeans are primarily used as cattle, swine, and chicken feed but is also consumed by humans in soy products and as food additives. In 2007, 216 million tonnes of soybeans were produced worldwide mainly in the US (33%), Brazil (27%), Argentina (21%), and China (7%). India and Paraguay also produce a respectable amount of soybeans. Almost 60% of the soybeans produced in 2007 were GM. The first GM soybeans were grown in the US in 1996 and is now grown in nine countries containing a herbicide tolerant trait. WHERE ON SHELVES: soy oil, soy protein, soy lecithin (used as emulsifier in chocolate, ice cream, margarine, baked goods), tofu, soy beverages, soy puddings, eggs, milk, meat

Sugar beet Sugar beets grow very slowly and are often outcompeted by weeds for light, water, and nutrients. Compared to other crops, they require the most frequent use of pesticides. Without the use of pesticides, only 25% of the crop would be harvestable and with cheap cane sugar outcompeting sugar beet sugar in the market, GM sugar beets became desirable. The German seed company KWS Saat AG developed the GM sugar beet H7-1 with the concept used by Monsanto on soybeans and other crops. The sugar beet is made resistant to the chemical glyphosate that stops the production of specific amino acids in plants, killing them. WHERE ON SHELVES: sugar, other processed foods

Cotton (imported) Cotton comes from the seed of a plant called Gossypium hirsutum and is grown on five continents. It is not only depended on for the production of textiles, but for animal feed and ingredients in other processed foods as well. After harvesting the fibres, the seeds can be used to make cottonseed oil, margarine, protein preparations, cottonseed milk, and animal feed. GM cotton is currently being grown in many countries making up 43% (15 million hectares) of cotton worldwide in 2007. The majority of GM cotton is grown in India and the US but it is also in China, Argentina, South Africa, Australia, Mexico, and Columbia. GM cotton is made to be herbicide tolerant and insect resistant. Bt cotton is insect resistant, including a gene from a soil bacterium that produces insecticidal toxins to stop caterpillars such as the bollworm from getting into cotton bolls and reducing yield and quality. This allows cotton farmers to drastically decrease the amount of pesticides they use. WHERE ON SHELVES: cottonseed oil, vegetable oil

Papaya (imported) Papaya is grown in tropical countries where the Papaya RingspotVirus is a threat. In the 1980s, the University of Hawaii developed a GM papaya that was resistant to this virus by inserting viral genes encoding capsid proteins giving it an immune response to the infection. The first GM papayas were grown in Hawaii in 1999 and today, they are grown around the globe in tropical countries. They are approved for consumption in Canada and the US but not in the EU. WHERE ON SHELVES: papaya, fruit juices

Squash (imported) There are several types of GM squash that are resistant to viruses that create mosaic patterns on the plant and distort the fruit. WHERE ON SHELVES: zucchini, yellow crookneck, straightneck squash

Milk Products with rBST (imported) Recombinant bovine somatotrophin (rBST) is a synthetic growth hormone that is injected into cows to increase milk production. It is not approved for use in Canada but dairy products imported from other countries such as the US may contain rBST milk. WHERE ON SHELVES: milk powder, dairy desserts, dairy drinks

The Process OF GENETIC MODIFICATION

Isolation of the gene of interest from an organism such as Bacillus thuringiensis

Isolation of Gene of Interest Restriction Endonucleases Cuts DNA into fragments, isolating the desired gene Restriction endonucleases, also known as restriction enzymes, are molecular scissors that cut double stranded DNA at recognition sites. Different enzymes recognize different recognition sites which are often four to eight base pairs long and characterized by a complementary palidromic sequences. A complementary palidromic sequence is a sequence that reads the same in the 5' to 3' direction of both strands. Once the enzyme binds to the recognition site, it cuts the DNA with a hydrolysis reaction that breaks the phosphodiester bonds. This may cause the fragments to have sticky ends that have DNA nucleotides missing their complementary base pairs or blunt ends that are fully base paired. Restriction enzymes that produce sticky ends are more useful as the fragments can be joined to other fragments cut by the same enzyme through complementary base pairing. Another enzyme called DNA ligase is needed to reform the phosphodiester bonds however, the fragments being complementary base paired will be able to form hydrogen bonds. Restriction enzymes that recognize a six base pair sequence are most ideal. Restriction enzymes that recognize a longer base pair sequence have a lower frequency of cutting the DNA while a shorter base pair recognition site results in a higher frequency of cuts. This is because if a restriction

DNA is extracted from the organism containing the gene of interest

enzyme recognizes an eight base pair sequence, the probability of it finding the sequence is once (only one sequence) in 65536 nucleotides (four base pairs in a eight base pair sequence, 4x4x4x4x4x4x4x4). In contrast, if the recognition site is 4 base pairs, the probability of finding the sequence is once in 4096 nucleotides (4x4x4x4). Fewer cuts is useful when scientists want to isolate a piece of DNA that includes a whole gene so that the gene itself is not cut into fragments. However, too few cuts can result in fragments that are larger than desired. Thus, restriction enzymes that recognize a six base pair sequence create a moderate number of cuts and are optimal in many applications. Restriction enzymes were first studied in the 1960s and discovered in 1970. In the 1960s, scientists noticed that inserting the DNA from one strand of E. coli into another strand of E. coli resulted in the DNA being cut into fragments. Following this discovery, two enzymes were discovered that could cut DNA randomly. In 1970, Hamilton Smith studied how some bacteria were able to resist attacks from viruses finding out that injected viral DNA was cut into fragments while the bacteria's DNA remained functional. This research led to the discovery and isolation of HindII, proving the existence of site-specific enzymes. Currently, there are around 2500 restriction endonucleases with 200 different target sites that have been isolated from prokaryotes. However, only 200 of these restriction enzymes are available

Restriction enzymes are added to the DNA to cut the DNA into fragments, isolating the desired gene commercially for research. Methylases Methylases are enzymes found in both prokaryotes and eukaryotes. In prokaryotes, they add methyl groups to nucleotides in recognition sites of restriction endonucleases in order to stop them from cutting the DNA into fragments. This protects the DNA of the bacteria while still allowing the restriction endonucleases to destroy any foreign DNA such as from a bacteriophage virus by cutting it. Methylases are important in genetic engineering as they can be used when working with prokaryotes to stop a gene from being cut in unwanted locations. Methylases are less commonly found in eukaryotes where they play a role in transcription. DNA Ligase DNA ligase can join fragments of DNA that have been cut by the same restriction enzyme. If sticky ends were produced, the DNA nucleotides missing their complementary pairs in both fragments will be attracted to one another, forming hydrogen bonds. DNA ligase joins the two DNA fragments by using a condensation reaction to reform the phosphodiester bond in the DNA backbone. However, in the case of joining two blunt ends together, DNA ligase is ineffective and enzyme called T4 DNA ligase is needed. T4 DNA ligase is isolated from T4 bacteriophages and requires ATP instead of NAD as a cofactor unlike DNA ligase.

Gel electrophoresis isolates the desired gene from remaining fragments of DNA

Gel Electrophoresis Separates isolated gene from remaining unwanted fragments of DNA After the desired gene has been isolated from the DNA, gel electrophoresis is a process used to remove remaining unwanted fragments using the chemical and physical properties of DNA. DNA is negatively charged with each nucleotide having a phosphate group that carries a -1 charge. The molar masses of each nucleotide are relatively constant with only a small variation due to the different ratio of purines to pyrimidines in each DNA molecule. This means that each nucleotide will have the same ratio between their charge and mass and the only difference between fragments of DNA are their lengths and different number of nucleotides. This allows for gel electrophoresis to separate DNA fragments based on size. First, DNA is placed in a well in a gel consisting of a buffer containing electrolytes and agarose. The DNA fragments to be separated are often mixed with glycerol, a heavy molecule that will cause the DNA to sink to the bottom of the well and allow it to be seen when migrating across the gel. A negative charge is placed on the end with the wells and a positive charge is placed on the other end. The negatively charged DNA fragments will migrate towards the positive side at a rate that is inversely proportional to the logarithm of their size. The shorter the fragment is, the faster it will be able to navigate through the gel and move towards to positive side. Before the shorter fragments reach the positive side, the

electrical current is turned off. Afterwards, the gel is stained with a stain such as ethidium bromide which can insert itself in the nucleotides of the DNA fragments, fluorescing them under UV light. DNA markers of known size put in one of the other wells can allow for comparison between them to determine the length of the fragments and the corresponding gene. A graph of the log of the known fragments size and distance travelled from the wells can also allow for estimation of the size of fragments through interpolation. In addition, the DNA fragments can be cut out of the gel for further purification and testing. Restriction Fragment Length Polymorphism (RFLP) Reveals small genetic differences in DNA if there are too many fragments of a similar size during gel electrophoresis Polymorphism is any difference in a DNA sequence, both coding and noncoding regions, that can be detected between individuals. As organisms of the same species carry the same genes but in different alleles, the genomes differ and are polymorphic unless they are identical twins. These differences are used as markers in the genetic mapping of various organisms. Restriction fragment length polymorphism (RFLP) analysis is the comparison of different lengths of DNA fragments produced by restriction enzyme digestion to determine genetic differences between the individuals by subjection to complementary radioactive probes. The concerned DNA is first digested by one or a combination of multiple restriction endonucleases. If gel

Restriction Fragment Length Polymorphism is used in cases where there are many fragments of the same size electrophoresis is used at this point, it would prove useless as the mass amount of DNA fragments only varying slightly in size would not be distinguishable on a gel. In this case, the gel is subjected to a chemical that denatures double stranded DNA into single strands. A process called Southern blotting is then used where an electric current transfers the single stranded DNA onto a nylon membrane. The nylon membrane is placed on a gel with a positive electrode behind it so that DNA, being negatively charged, will transfer out of the gel and onto the nylon. The nylon is then placed in a solution containing radioactive complementary nucleotide probes for desired regions. Hybridization will occur with the complementary base pairing between the DNA and probes. When placed against an x-ray film, an autoradiogram will be produced where areas where the radioactive probes have bonded will cause the film to become exposed. By developing the film, the areas will become visible and the differences in patterns can be analyzed.

The gene of interest is inserted into plasmids to be cloned

Cloning of Gene of Interest Plasmids Expresses inserted gene and clones it To have an isolated gene be expressed as a useful protein, they are inserted into the plasmids of bacteria. Plasmids are small, circular, double stranded DNA molecules without a protein coat found in the cytoplasm of bacteria. They are independent of the chromosome in the bacteria cell and range from 1000200,000 base pairs. Plasmids often carry genes that offer antibiotic resistance, protection to heavy metals such as mercury and lead, and the ability to decompose herbicides, industrial waste, or components of petroleum. This relationship between bacteria and plasmids is endosymbiotic meaning both benefit from the mutual arrangement. Plasmids also contain a copy number that indicates the number of copies of a specific plasmid found in a bacterial cell. The greater number of copies there are, the greater the influence of the gene. Artificial plasmids have been engineered to contain a multiple-cloning site where the recognition sites of many restriction enzymes are all contained, allowing for several cuts in the one region. The recognition site of one restriction enzyme is only found once in the plasmid so only one cut it made and the plasmid is not cut into fragments by the same enzyme. One cut in the plasmid will make it linear and if the foreign gene has been cut with the same restriction enzyme, both fragments will have the same complementary base ends resulting in hydrogen bonding between the two. The gene of interest must be placed following a promoter which is the recognition site during protein synthesis. This way, during protein synthesis, the foreign gene will be synthesized along with the original gene. If the foreign gene must be regulated, it is often placed with an inducible promoter

The Polymerase Chain Reaction can be used as an alternative to plasmids to clone the DNA of the gene

so that it is only synthesized when the corresponding substance is present and needs to be broken down. DNA ligase is then added to reform the phosphodiester bonds resulting in a circular plasmid again. This combination is known as recombinant DNA as it now contains original and foreign DNA. When it is inserted back into the bacterial cell, the gene will be cloned as the plasmid containing the foreign DNA replicates to form more copies. Polymerase Chain Reaction (PCR) Another method of replicating DNA besides the use of plasmids The polymerase chain reaction (PCR) developed in 1987 by Kary Mullis is an advanced technique used in biotechnology that allows for a desired gene to be copied without inserting it into a plasmid. PCR is often used in place of plasmids because it is a more direct method of copying DNA without having to be in question of whether the transformation was successful. In PCR, the strands of DNA are separated by subjecting them to high temperatures of 94˚C -96˚C in order to break the hydrogen bonds. One DNA primer complementary to one end of a DNA strand is used while another complementary primer of the opposite end of the opposing strand is used. The DNA primers, being complementary to the 3'-5' ends of the template end, are 5'-3' primers. One is known as the forward primer while the other is a reverse primer as they initiate DNA synthesis in opposite directions of each other. Afterwards, the temperature is lowered to 50˚C -65˚C to allow the primers to better attach to the template DNA. Taq polymerase, a DNA polymerase, then builds the complementary strands using the free nucleotides added to the solution at a temperature of 72˚C. Taq polymerase originates from the bacterium Thermus aquaticus that lives in hot springs, therefore

The DNA of the gene is now purified and there is an adequate amount to insert into the organism of interest

its enzymes has the ability to withstand high temperatures. When the complementary strands have been built, the cycle is repeated with the number of copies doubling each time resulting in an exponential growth. During the first few cycles of the PCR, Taq polymerase replicates the DNA from the beginning to end regardless if it is the desired target area or not. To discontinue this, after the first cycle, variable-length strands are produced that start and end in accordance with the target area. During future cycles, on the two strands of DNA, one end will terminate at the target region where the variable-length strand was produced in the previous cycle, and a primer will attach to the other end of the target region. Taq polymerase will then only replicate from the primer to the terminator at the end of the target region. The strands produced are called constant-length strands. The remaining two strands are extended by Taq polymerase like in the first cycle so that by the third cycle, the number of strands consisting of only the targeted area will increase exponentially.

The Ti plasmid vector enters through the wound of a plant, creating a crown gall

Induction of Transgenics Transformation Introduces foreign DNA by plasmid or virus into another cell Transformation is the introduction of DNA from one source into another organism. Plasmids are often the organism that inherits the new DNA, becoming transformed. By doing so, they can act as vectors, carrying a desired gene into a host cell. A competent cell accepts foreign DNA without any problems, however many bacterium are not competent and must be manipulated in the laboratory with the help of the salt calcium chloride that ionizes into Ca2+ and Cl-. The bacteria is placed in a solution of calcium chloride at 0Ë&#x161;C, where the bacterial membrane's negatively charged phosphates are stabilized by the positively charged calcium ions. In addition, the low temperature freezes the fluid cell membrane establishing a more stable chemical and physical environment. At this time, the plasmid DNA is added to the solution and as the phosphate groups hold a negative charge, they are also stabilized by the positively charged calcium ions. Afterwards, the solution is heated to 42Ë&#x161;C for 90 seconds to create a draft. The outer environment of the cells being at a slightly higher temperature than the inside allows the plasmids to enter the bacterial cell through the pores of the membrane. The solution is then left to settle in an incubator at 37Ë&#x161;C in a nutrient rich medium. Selective plating is conducted to isolate cells that have successfully transformed with recombinant DNA from those that failed. An antibiotic-resistant gene is often added to the plasmid before it is reinserted into the bacteria so if the transformation was a success, it will be able to grow on medium containing an antibiotic. If there is no growth on the medium, then both the foreign gene and antibiotic gene were not successfully inserted, disabling it

A gene gun shoots particles covered in DNA into a plant

The plant regenerates on a growth medium before being transferred into soil

from surviving in an antibiotic environment. In some instances, the foreign gene fails but the antibiotic gene is successful in the transformation. This makes it necessary to test colonies of bacteria that have transformed. A successful colony is isolated and allowed to colonize until there is enough to extract plasmid DNA from it. The plasmid DNA is then treated with the same restriction enzyme used when creating the recombinant DNA so that the cloned inserted gene will be excised. If the gel shows the expected pattern of bands, the colony does carry a recombinant DNA plasmid. In addition to the calcium chloride method, a more modern technique for transforming cells is with the use of electroporators that subject the bacteria to an electric shock. The electric shock makes the cell wall temporarily more permeable so that foreign DNA can enter. In the case of animal or rigid plant cells, other methods are used. Methods of Gene Transfer in Plants The Ti Plasmid- Dicotyledons In 1981, Eugene Nestor and Mary Dell Chilton created a method to insert foreign genes into plants cells to produce transgenic plants. They used the tumourinducing (Ti) plasmid found in the soil bacteria Agrobacterium tumefaciens as a vector. The bacteria, being attracted to the chemicals released in the wound of a plant, can easily enter through the wound. The Ti plamid enters the plant cells through natural transformation and causes a tumor-like growth on the plant called a crown gall. The bacteria takes nutrients from the plant while the plant does not benefit in any way. The T region in the Ti plasmid is the only region that gets inserted into the chromosome of the plant cell, thus foreign genes are inserted in this region of the plasmid. The infected tissue is then extracted and cultured on a shoot-inducing medium for 2-3

days allowing for the transfer of the foreign gene. Afterwards, they are transferred onto a medium containing antibiotics to remove the tissues that have not transformed and then in about 3-5 weeks, a root-inducing medium. In 3-4 weeks, the roots will be formed and the plants can be transferred to soil. Ti plasmids can only be introduced into dicotyldons such as beans, peas, and potatoes. The Gene Gun- Monocotyledons The method of using a gene or particle gun, referred to as biolistics, was invented in 1988 by John Sanford. 1-3 micrometers sized gold or tungsten particles are coated in the foreign DNA and loaded into a electrical gene gun. The gene gun is then bombarded at the plant, allowing the DNA to penetrate the cell wall and membrane. The cells are then placed on a growth medium to develop and are subjected to antibiotics to confirm the success of the transformation. This method is commonly used on primary explants and proliferating embryonic tissues to introduce genes into monocotyledons such as wheat, corn, and rice, a gene gun is needed. Gene Expression In addition to testing the plant for resistance against the specific antibiotic for the success of transformation, they must be assessed for the expected traits of the gene as it is not necessarily expressed even if successfully integrated. Plants are also observed for the number of copies of the genes inherited and the new genes influence on existing genes. The plant performance is first assessed in a greenhouse and then in the outdoors. If the desired trait is expressed and there are no unwanted characteristics, the new plant may be considered for commercial use.

GM crops are often created to include genes making them pesticide resistant, virus resistant, herbicide tolerant, environmental tolerant (drought, heat, salinity, more nutritional, or to stay fresh longer. One of the first GM crops made included a gene that gave the plant resistance to certain pests. A type of pesticide resistant gene commonly used is found in the soil bacterium Bacillus thuringiensis (Bt) which produces a selective toxin. Isolated and inserted into crops such as corn, cabbage, and cotton, these plants will be able to secrete their own toxins killing off specific pests such as caterpillars. Herbicide resistant plants give farmers the ability to spray chemicals that are efficient in killing all the weeds without harming their own crop. Herbicide tolerant crops such as soybeans are commonly engineered to be resistant to glyphosphate, a widely used, nonselective herbicide that kills all plants it comes in contact with without remaining in the soil. Before, glyphosphate could not be used and only expensive, ineffective, selective pesticides that remained in the soil could be used without killing all the plants including the crop. However, with herbicide tolerant crops, broad spectrum herbicides can be used without killing the crop, offering economical and environmental benefits. Crops are also often modified to have an extended shelf life. This can be done by slowing down the ripening process by manipulating the enzyme pectinase which is responsible for breaking down the pectin holding plant cells firmly together. One of the first crops altered to have a longer shelf life was the Flavr Savr Tomato where the gene responsible was removed and reinserted inversed to decrease the amount of pectinase produced, slowing down the ripening process and increasing the shelf life.

Crops being Developed around the World Crop Apple Banana Broccoli Cabbage Celery Coffee

Trait (not necessarily all in the same plant) Insect resistant Free of viruses and worm parasites Delay ripening so remains green longer Resistant to caterpillars Stays crisp longer Better flavour and yield, pest resistant, decrease caffeine content

Cucumber Melon Potato

Resistant to viruses, fungi, bacteria Extend shelf life Resistant to caterpillars and beetles, decrease need for fertilizer, decrease water content for low-fat chips

Raspberry Strawberry

Increase sugar content, extend shelf life Frost resistance for early season production Increase nutritional oils, decrease saturated fat content

Sunflower Tomato

Wheat

Resistant to viruses and frost, increase yield, delayed ripening, increase sugar content, decrease fat content, salt resistant Herbicide resistant, increase flour suitability for bread making,

Bt Corn influence as organisms in the soil are commonly exposed to this toxin. The amount of endotoxins released are also 10 times lower than the level at which there are observable effects to soil organisms such as earthworms and springtails. Delta endotoxins and VIPs are not considered potential food allergens as they are rapidly broken down through digestion. In tests, they have been subjected to acidic environments where the survival time is not long enough to likely survive the digestion process and be absorbed by the bloodstream in humans. There has been an Ontario study where delta endotoxin Cry1Ab was detected in the blood serum of pregnant woman, fetuses, and non-pregnant women but the health effects to this are unknown. Another concern is the ability of Bt corn pollen to contaminate non-Bt corn crops which is an issue to international trade and certified organic producers. Monarch Butterflies Monarch caterpillars feed on milkweed and in tests where they were fed milkweed covered in Bt corn pollen, they died. In addition to this, monarch butterflies tend to lay eggs on milkweed plants near cornfields whether they are growing nonBt or Bt corn. However, studies show that the survival rate of monarch caterpillars is the same in Bt cornfields and non-Bt cornfields. In some cases, they have even shown a higher rate of survival in Bt cornfields as they are untreated with other insecticides. The lethal dose (LD50) of Bt corn pollen to monarch caterpillars is 2500 pollen grains/in2 of milkweed leaf (produced by event 176 which is no longer available for commercial use, producing 15-25 times more delta endotoxin containing pollen grain than the events still in use). At 850 grains/in2 there appears to be no effects and of the events still in use, no known effects are being observed. Studies also show that rainfall helps reduce the amount of pollen on milkweed plants and there is low overlap between pollen shed and the presence of monarch caterpillars. However, depending on the weather and location of different cornfields, there may be areas where these events happen simultaneously.

105 records of individual plot yield records found for Monsantoâ&#x20AC;&#x2122;s DEKALB corn in 2014 in Ontario.

Bt corn is corn that is genetically modified to include a gene from Bacillus thuringeiensis (Bt), a soil bacterium that produces insecticidal toxins. This introduced gene makes corn resistant to certain pests such as lepidopteron caterpillars, in particular the European corn borer, by producing the pesticide in its pollen. There are no known adverse human affects however, non-target insects closely related to the target insect may be affected. Delta endotoxins and vegetative insecticidal proteins (VIPs) are toxins released by Bt corn. They are considered safer for humans and non-target pests than regular insecticides. This is because they are more selective with the toxins released attacking sites that are only found in some groups of insects. Delta endotoxins and VIPs are classified as Generally Regarded as Safe (GRAS) by the US Environmental Protection Agency (EPA) and are approved for most organic certification programs. Studies have shown that the entire Bt corn plant may be toxic with the level of toxicity depending on two factors, the event and the promoter. Different seed companies use different events and promoters with their hybrids varying in what plant tissue the Bt toxins are produced in. Event The insertion event is the event of placing the Bt gene into the DNA of the corn plant. This will determine the physical location of the gene such as the chromosome and part of chromosome which dictates where and the amount of plant toxins produced. There is currently no technology to control the Bt gene location with each event leading to plants that vary in where and how much toxin is produced. Promoter The promoter is the genetic switch that controls where and when the Bt gene produces toxins. There are several different promoters available on the market that each affect the location and amount of toxin produced differently. Concerns Studies have shown that the roots of Bt corn plants leak delta endotoxins into the soil. However, it is believed to be of no

Pollen grains/in2 of milkweed leaf Lethal dose (event no longer produced)

2500

No observed effects

850

Within Bt cornfields

500

Near Bt cornfieldswithin 3 feet

180

Near Bt cornfieldswithin 5 feet

THE FLAVRSAVR TOMATO ~ The Flavr Savr Tomato was the first genetically modified crop to be sold commercially. The research and marketing done created a scientific and temporary sales success.

The Flavr Savr Tomato first genetically modified crop to be sold commercially

Animal testing was conducted by Calgene, who concluded that the PG antisense tomato was the same as the regular tomato in all ways besides the fruit cell wall pectin degrading more slowly and the tomato paste having a higher viscosity. On May 21, 1994, the Flavr Savr Tomato was introduced to the public. The demand was high, however Calgene was not able to make any profit with the high cost of production and distribution. In 1996, Zeneca introduced their product of tomato paste made from PG antisense tomatoes grown and processed in California to the United Kingdom, under licence. Zeneca worked with Sainsbury's and Safeway, selling 1.8 million cans from 1996-1999 with the can label stating the paste was derived from GM tomatoes. The GM tomato paste outsold regular tomato pastes, however sales started declining in 1998. Soon after, Sainsbury's and Safeway stopped carrying the GM products with the explanation of

addressing customer concerns rather than food safety. This decline in sales can be traced to the Bristish Broadcast in August 1998 where Dr. Arpad Pusztai announced that feeding rats GM potatoes lead to biological effects as a result of the process of genetically engineering rather than the product of the introduced gene. However, he later concluded that the statements made in the broadcast were incorrect. However, the market of Zeneca's GM tomato paste did not resume. MORE ON THE GM TOMATO TEST The test Calgene conducted was not reviewed by others or published but can be found on the internet. The test concluded that toxic effects were absent as well as that body and organ weights, weight gain, food consumption, and clinical chemistry or blood parameters were insignificantly different from the rats on regular diets. However, there were many flaws in the test that prove these results invalid. Firstly, the starting mass of the rats ranged greatly by 1823%. Also, histology was not conducted on intestines although lesions were found in the stomachs of 7/20 the female rats while none were found in the control group. In addition, 7/40 of the rats fed GM tomatoes died within 2 weeks for unexplained reasons. This study was poorly designed with the safety of the Flavr Savr Tomato and the decision of discontinuing toxicological testing on other GM foods in the future by the FDA in question. Athanasios Theologis holding a tomato with the gene responsible for ripening blocked.

n the 1980s, researchers at Calgene Inc. saw that the tomato enzyme polygatacturonase (PG) dissolved cell wall pectin softening the fruit. They looked to suppress PG formation in ripening tomatoes by introducing the antisense, reverse orientation copy of the gene. With this, the ripe fruit would stay firm longer allowing the transport of vine-ripened fruit as opposed to picking unripe fruit to be transported and then artificially ripening them with ethylene. In 1987, the tomato PG gene was cloned, methods for the transformation and regeneration of the tomato were planned, and the GM tomatoes were grown. The resulting tomatoes produced as low as 1% of PG as regular tomatoes. In 1992, the US Department of Agriculture determined that the PG antisense tomato lines were not a risk removing the requirement of permits for field testing and transportation. Later, the US Food and Drug Administration approved the introduction of kanamycin-resistance gene constructions to make PG anti-sense gene tomatoes in 1994. Kanamycin-resistant organisms being abundant in human and animal guts as well as the soil, would not cause any influence or allergic reactions in the genetically engineering tomato gene.

Salt Resistant Tomatoes New modifications to the tomato are always in research including the salt resistant tomato created by scientists from the University of California in 2001. 24.7 million acres of farmland worldwide are lost each year as a result of the salinity of modern irritation methods. The new salt resistant tomato would be able to withstand soil that is 50 times saltier and be grown in areas other crops would not survive in. These tomato plants transport salt to their leaves, leaving the taste of the fruit unaffected. This process will also clean the soil, removing salt build up. This concept is planned to be used in other plants as well, although this salt resistant tomato is not yet approved for commercial agriculture.

Genetically modifying animals is more difficult than modifying bacteria or plants. To modify an animal to have improved performance, the changes must be made while it is still an embryo. Before the fertilized cell has divided, the desired gene must be injected into the nucleus of the egg using a precise tool such as a micropipette. However, this process has a low success rate with the DNA only being successfully transferred in about one of every hundred fertilized eggs injected. Animals that have been worked with include cows, pigs, chickens, sheep, and fish to have enhanced growth rates, leaner meat, and be disease resistance.

Cows & rBST Cows produce a form a growth hormone called bovine somatotrophin (BST). It has been found that by injecting excess BST into cows increase their muscle and milk production. BST was originally extracted from dead cows but can now be produced in bacteria with tools of genetic modification. Synthetic BST is called recombinant bovine somatotrophin (rBST). One dose of rBST every two weeks increases milk production by as much as 25%. However, the huge increase in energy needed for the increased production of milk often causes cows to become ill and more susceptible to udder and gut infections. As a result, cows are given antibiotics but their lifespan is often still shortened. rBST is not approved for use in Canada but since 1993, it has been legal for use in the US. In the US, milk produced for cows treated with rBST is not labelled as it is considered as safe as milk produced from cows without rBST treatment. Dairy products imported into Canada are likely to contain milk from rBST cows. There is no way to identify these products according to the Canadian Food Inspection Agency (CFIA) but it is possible to choose products that originate from countries that have not approved the use of rBST such as Canada. However, products where milk is a minor ingredient can still be classified as made in Canada regardless of where the raw ingredient originate from.

Growth Hormones and Antibiotics Hormonal growth promoters (HGPs) are used in beef cattle to increase the rate of production by increasing lean tissue growth and in turn decrease the cost. There are six HGPs approved for use in beef cattle in Canada. Progesterone, testosterone, and estradiol-17Ă&#x; are natural while trenbolone acetate (TBA), zeranol, and melengestrol acetate (MGA) are synthetic, all of which are given as subcutaneous implants (behind the ear) besides MGA which is added to feed. HGPs are only approved for use in beef cattle and not in any other livestock such as poultry or pork. Antibiotics are given to treat illnesses and prevent disease by killing or reducing harmful bacteria. Antibiotics are used on cattle, chickens, pigs, and fish raised for meat as well as fruit and honey bees. When animals that produce products such milk or eggs are on antibiotics, their products are discarded. The CFIA tests both domestic and imported food regularly to ensure they are below the level of hormones and antibiotics safe for humans stated by Health Canada. HGPs and antibiotics have been approved for use in Canada as no evidence of them being a threat to human health have been found. As for concerns of antibiotics creating resistant bacteria in humans, the government is keeping up to date with any scientific evidence produced.

GM Fish Salmon

GE salmon has not been approved for commercial production yet but it is on its way. There are a few variations of these salmon but they are all modified to reach maturity quicker. One variety contains a gene from a flounder fish that allows a growth rate 400-600% faster. They do not grow larger than normal salmon but mature over twice as fast in a 16-18 month period. These salmon also have a lower fat content which is appealing to some consumers. Some concerns of these salmon is that they will escape and breed with the wild population of salmon, outcompeting them. However, companies have countered that their salmon are female and sterile and will be raised on land based facilities to eliminate the possibility of escape.

AquAdvantage® Salmon (background) vs. a non-transgenic Atlantic salmon sibling (foreground) at the same age.

How AquAdvantage® Salmon Eggs are Produced

Aquabounty Technologies One company looking to introduce their GM salmon to the market is Aquabounty Technologies. Aquabounty believes that as approximately 85% of the worlds fish reserves are expected to become depleted when the demand for fish is increasing by 18.8% per capita consumption (14.4kg in the 1990s to 17.1kg in 2011), GE fish will become essential in the near future. Their product, AquAdvantage® Salmon (AAS) includes a gene from Chinook salmon giving it the potential to grow to market size in half the time of conventional salmon. The company claims that the AAS are identical to other Atlantic salmon in all other aspects. Aquabounty plans to grow their all-female, sterile salmon on land based facilities as to eliminate the risk of them escaping and crossbreeding with the wild population. Their AquAdvantage® Salmon eggs will give rise to fast growing fish that will help feed our overpopulated planet while reducing stress on wild fisheries. Other advantages include reducing the need for coastal areas, transfer of disease from farmed to wild fish, cost of feed, and being on land close to markets, the environmental impact of transportation. Aquabounty believes that AquAdvantage® Salmon is the future of salmon aquaculture.

The broodstock are carefully selected to produce the highest quality milt and eggs. Unfertilized eggs are massaged out of female non-transgenic salmon.

Milt is taken from male transgenic AquAdvantage® Salmon. The transgene in their genome is passed down naturally from generation to generation. Aquabounty has produced ten generations of AquAdvantage® Salmon at their hatchery in Canada.

The combination of non-transgenic eggs and transgenic milt produces fertilized eggs.

The fertilized AquAdvantage® Salmon eggs are pressure shocked so the fish hatched from the eggs are sterile and unable to reproduce.

The fertilized, sterile, all-female AquAdvantage® Salmon eggs are placed in incubators until they become eyed eggs so they can be shipped.

The History of GMOs 1970

1953

Hamilton Smith discovers the first restriction enzyme, HindII

James Watson and Francis Crick discover the double helix structure of DNA

1950s

1960s

1973 Herbert Boyer and Stanley Cohen combine their knowledge on restriction enzymes and plasmids to create the first recombinant DNA organism

1970s 1972

1975

Janet Mertz and Ron Davis produce the first recombinant DNA using restriction enzymes

Asilomar Conference held in the US to discuss guidelines on using the newly discovered recombinant DNA technology

The Structure of DNA DNA, deoxyribonucleic acid, is a double stranded polymer of nucleotides (each consisting of a deoxyribose sugar, phosphate group, and one of four nitrogenous bases) that carries the genetic information of an organism. Deoxyribose is a five carbon cyclic ring structure that contains four carbons and an oxygen atom plus another carbon extending out from the ring. The carbons are numbers clockwise starting from the carbon right of the oxygen which is numbered 1 prime or 1'. The nitrogenous bases are attached to the 1' carbon by a glycosyl bond while the phosphate group is attached to the 5' carbon (and to the 3' of the next sugar) by a phosphodiester bond. The four nitrogenous bases are adenine and guanine which are double ringed structures referred to as purines and thymine and cytosine which are single ringed structures referred to as pyrimidines. The total amount of adenines always equal the amount of thymines and the total amount of guanines always equals the amount of cytosines present in a DNA molecule. The total amount of purines and pyrimidines are also always equal. DNA consists of two anti-parallel strands of nucleotides with one strand running in the 5' to 3' direction and the other in the 3' to 5' direction. The bases of one strand pairs with the bases in the other strand turned inwards, forming hydrogen bonds. A purine must always be paired with a pyrimidine, however the only two possible complementary pairs are adenine paired with thymine and guanine paired with cytosine. The diameter of DNA is a constant of 2nm (nanometers) and the double helix turns in the clockwise direction making a full turn every 10 nucleotides to account for the 3.4nm full helical twist.

1983

1980 The US Supreme Court rules that GMOs can be patented

Kary Mullis creates the polymerase chain reaction which allows DNA replication to proceed under an accelerated rate ~ Luis Herrera-Estrella creates the first GM plant by using Agrobacterium to transfer an antibiotic resistant gene into a tobacco plant

1980s 1982 FDA approves first GE product, Genentech's Humulin, a type of human insulin produced by bacteria ~ First transgenic mouse born

1990

1996

First GMO used in food, a modified yeast for bread making

First GM soybeans and corn planted in North America

1993 The FDA approves Bovine somatotropin (bST), a metabolic protein hormone used to increase milk production in cows

1990s 1987 Monsanto announces its success in creating the GM insect resistant crop

2003 Human genome sequenced

2000s 1992

1994

The FDA declare GMOs do not require special regulation

Calgene's Flavr Savr tomato sold commercially

The Asilomar Conference The Asilomar Conderence held in California included scientists from around the globe, government officials, and members of the press. They discussed concerns of the newly discovered recombinant DNA technology giving them the power to clone DNA from any organism on the planet. The group acknowledged that the new technology would lead new advancements in medicine, agriculture, and the industry but also posed risks to human health and the environment. The validity of the moratorium placed after discovering the new technology was discussed. The conference ended in lifting the moratorium despite a number of opposes and guidelines on how experiments should proceed were set. The conditions mainly consisted of protecting laboratory personnel, the general public, and the environment from hazards resulting from experiments. During the time, many had fears of common bacteria becoming pathogens with the insertion of genes giving them the ability to become antibiotic resistant, produce toxins, or act as cancer causing agents. Many also believed lifting the ban was a mistake and proceeding in the use of recombinant DNA would result in a disaster. Since 1975, millions of experiments have be performed without any of these fears coming true or posing any major threat to the public. Attendants and the public concluded that the conference was a success with scientists taking the initiative of raising the issue themselves rather than having the issue raised on them.

The First Transgenic Mouse The first transgenic mouse was created by transferring a gene from one animal to the embryo of another effectively so that the gene would be expressed in the mouse and its offspring. This experiment was conducted by Richard Palmiter and Ralph Brinster. They were looking to understand more on how cells read DNA and translate it into biological structures. The technique they imposed was based on the fact that genes consist of two parts, one that codes for specific proteins, and one that determines when to activate the gene. Today, it is known that these parts are called coding regions (there are also non-coding regions) and the promoter. Palmiter and Brinster placed a gene that produces the enzyme thymidine kinase with the metallothionein (MT) promoter which is responsible for activating the production of MT which binds the metal ions of copper, zinc, and cadmium. Placing the gene into the fertilized eggs, they soon found adding cadmium also produced thymidine kinase. In a different experiment, they added growth hormone with the MT promoter, producing abnormally large mice. The work of Palmiter and Brinster provided a way of testing experimental models and treatments for disease. In one experiment, mice were given a gene that lead to non-functioning livers and then given healthy liver cells, resulting in new, healthy liver tissue. Other diseases that have been studied include cancer, sickle cell disease, and diabetes.

Concerns Dr. Arpad Pusztai’s Research Dr. Arpad Pusztai and a team of researchers at the Rowett Institute of Scotland received a million dollar grant to study the potential health problems of GE foods including potatoes. His results were all over the British media in 1998-1999. However, a short while after he publicized his concerns, he was fired, locked out of his lab and his research and funding ended. From the research he had completed, he found: •significant chemical and compositional differences between conventional and GE foods •significant nutritional and protein difference between batches of the same GE food, a result of random gene insertion •only after ten days, damage to vital organs and immune systems of lab animals fed GM potatoes •rats fed potatoes with lectin added genetically (GE) were harmed while rats fed potatoes with lectin added mechanically (eyedropper) were unharmed, suggests harm from impact of GE process and not foreign gene •Cauliflower Mosaic Viral Promoter (CaMv) inserted into most GM foods harmed lab animals

Antibiotic Resistance The antibiotic resistant marker gene inserted along with foreign genes in our GM food have the potential of mating with pre-existing bacteria in our gut, and for up to an hour after ingestion, in our mouth and throat. This could give rise to new deadly strains of antibiotic "superbugs" such as salmonella, E. coli, tuberculosis, and staph that cannot be cured by traditional antibiotics such as ampicillin, kanamycin, and penicillin. This transfer of antibiotic genes into unintended organisms is an example of horizontal gene transfer that up until GE, had been very rare.

Bovine Growth Hormone & Cancer Common in dairy practices in the US is the GE hormone recombinant Bovine Growth Hormone (rBGH), also known as recombinant Bovine Somatotrophin (rBST) which was approved for sale by the FDA in 1994. Found in the milk produced from cows injected with rBGH is a high level of the chemical hormone Insulin-Like Growth Factor (IGF1) that increases the risk of breast, prostate, and colon cancer. Compared to natural milk, the level of IGF-1 in GE milk and dairy products is ten times higher. This powerful cancer tumour promoter resists pasteurization and digestion in the stomach and is absorbed by the intestinal walls. In addition, cows on rBGH have been shown to have a higher risk of mastitis, an udder infection that must be treated with antibiotics that can end up as residue in milk and beef. rBGH milk may also contain a higher level of pus and bacteria in the milk.

GMO Testing There are many opinions on GMOs ___but still limited data on the health _risks. These uncertainties should have been tested for before allowing humans to consume GM foods. Humans have become part of the experiment with potential health concerns in the future. The few scientific studies that have been published are flawed and were conducted with low standards. In many cases, the GM and parental lines are not equivalent and although some argue this does not concern us biologically, the differences in composition, nutrition, toxicity, and metabolic differences are evident. A more solid scientific foundation should be laid for GMOs before they become more depended on in our future.

Allergenicity Testing Another health concern of GM foods are their influence on allergies and anaphylaxis in humans. However, this uncertainty is difficult to assess as there are good animals for nutritional and toxicological testing but no animal can fully demonstrate the effects of GM foods on allergies in humans. Testing GM foods _in relation to allergic reactions is further __complicated to conduct if the gene ____transferred is from an organism that _____has not been consumed before.

Regulations In Canada, Health Canada is responsible in overseeing matters related to public health, food safety, and nutrition. Biotechnology is a widely used term that includes foods produced from the use of scientific tools and techniques. These foods are commonly called genetically engineered or genetically modified foods and are considered novel foods. It takes 7-10 years to assess the safety of a new GM food before introducing it to the market in Canada. Companies that look to sell new GM foods must first submit data to Health Canada for a pre-market safety assessment (as under Division 28 of Part B of the Food and Drugs Regulations, Novel Foods). This process can be summarized into 8 steps. 1. Pre-submission consultation Health Canada encourages consulting the Novel Foods Section of the Food Directorate prior to proposing a GM food for safety assessment to clarify any questions on the regulatory process requirements. 2. Pre-market notification A submission is made to Health Canada which is fully assessed by scientific evaluators. The GM food must meet all criteria under the Health Canada's Guidelines for the Safety Assessment of Novel Foods. 3. Scientific assessment Further assessment of the product is made including the development, composition of nutritional content, potential for production of toxins, potential of causing allergic reactions, chemical safety, potential secondary effects, and both major and minor constituents. 4. Requests for additional information If insufficient information was submitted, further documentation is requested. 5. Summary report of findings Scientific evaluators summarize their findings in a report. 6. Preparation of food rulings proposal A Health Canada Food Rulings Proposal is prepared and reviewed by the Directors and Director General in the Food Directorate. The decision of whether to approve or decline the introduction of the GM food is made. 7. Letter of no objection If the product is approved, a Letter of No Objection is sent to the company stating that the product can be sold in Canada. 8. Decision document on Health Canada website The decision and information on the novel food and evaluation results are posted on the Novel Foods and Ingredients page of Health Canada's website.

Food Labelling Food labelling policies under the Food and Drugs Act is the responsibility of Health Canada and the Canadian Food Inspection Agency (CFIA). Health Canada safeguards health and safety while CFIA develops general food labelling policies and regulations. Foods that pose a health or safety issue such as having a different nutritional value or composition or containing an allergen must be labelled. In regards to GM foods, labelling is voluntary as summarized in the Canadian Standards Voluntary Labelling and Advertising of Foods that Are and Are Not Products of Genetic Engineering which looks at addressing issues rather than labelling. The group responsible includes consumer groups, food companies, producers, interest groups, universities, and the government with the objective of providing meaningful, understandable, and truthful labels.

GMO Regulation Around the World

Current Issues GM Potatoes In November 2014, the US government approved the “Innate” GM potato by Simplot for the commercial market. This potato is modified to be bruise resistant and reduce asparagine, an amino acid that reacts with specific sugars to oxidize into acrylamide, a carcinogen, at 120ºF (49ºC) which is reached during deep frying. Since 2001, this is the first GM potato that has been proposed after Monsanto’s GM insect-resistant potato being protested off the market.

GM Fish In November 2013, Environment Canada approved the commercial production of GM Atlantic salmon eggs. Aquabounty Technologies is now awaiting approval of GM Atlantic salmon for human consumption in the US and Canada. If these GM fish are approved, they will be the first GM animals for food in the world.

GM Alfalfa In 2014, CBAN stopped GM alfalfa from being approved but it may proceed in 2015. Alfalfa is harvested for hay for animal feed (dairy cows, beef cattle, poultry, pigs, lambs) as it is high in protein and builds up nutrients in the soil, which is especially important for organic farming. Farmers are against GM alfalfa and have been protesting against its approval for five years. If Monsanto’s GM herbicide tolerant (Roundup Ready) alfalfa were introduced, both organic farming and conventional farming would be at threat. GM contamination would also be probable as alfalfa is a perennial crop pollinated by bees.

GM Apples Okanagan Specialty Fruits is a small Canadian company that is awaiting approval of its GM "non-browning" apple in Canada and the US. The approval would be an issue for apple farmers and consumers.

Low Level Presence As of July 2014, the Canadian government is looking at legalizing the presence of unapproved GM foods at 0.1% or lower in our food. These GM foods do not have to be approved for safety in any other country as the government argues that this low level is not harmful. Visit http://www.cban.ca/Take-Action to learn more

The top three GM crops (measured by the amount of land used in 2009) are soybeans (52%), maize (31%), and cotton (12%). The remaining include canola (5%), sugar beet, alfalfa, and papaya.

Past Issues FlavrSavr Tomato Calgene's Flavr Savr tomato, the first GM food to be sold in the US in 1994, was a temporary sales success having a long, attractive shelf life. These tomatoes and its products outcompeted conventional tomatoes in supermarkets until Dr. Arpad Pusztai publicized his research on GM potatoes. In his research, lab rats fed GM potatoes developed lesions in their stomachs and died. Afterwards, the public boycotted these tomatoes, putting the company out of business.

NewLeaf Potato Monsanto's NewLeaf potato containing a gene to make it pesticide resistant to the Colorado potato beetle was introduced to the market in 1995. This potato was expected to be a huge success, especially in the fast food and junk food market, but in 1998 the Institute of Nutrition released a study with results of rats developing organ damage after being fed NewLeaf potatoes. In 1999, farmers were planting over 50,000 acres of NewLeaf potato but by 2001, the product was out of the market.

Monsanto is the largest GM seed company owning over 86% of GM seeds sown globally. The biotechnology company puts small farmers out of business, has multiple connections to the government, and has a long history of many chemicals including PCBâ&#x20AC;&#x2122;s, dioxin, and agent orange.

Starlink Corn In 1998, Starlink corn was released with a gene making the plant pesticide resistant. This corn was limited to be used in animal feed and fuel as it caused allergic reactions in humans. However, Starlink Corn contaminated other varieties of corn grown for human consumption, making its way into the food supply. This contamination caused production to be shut down, however, Starlink Corn DNA has still been recently found in August 2013 in the Saudi Arabian food supply.

LibertyLink Rice In 2011, LibertyLink rice with the trait of being tolerant to glufosinate, the active ingredient in Liberty herbicide, contaminated the conventional long grain rice of thousands of US farmers. As a result, Bayer AG had to pay $750 million as compensation, however, an entire strain of rice was lost permanently.

GM Wheat

Globally, 79% of soybean s, 70% of cotton, and 32% of maize are now GM crops.

In 2002, Monsanto proposed GM wheat with the same herbicide tolerant gene in their other crops. Many farmers were not interested with the fear that GM wheat would contaminate natural wheat, destroying complete harvests. At one point an unapproved GM wheat strain was found in an Oregon field, making Americans suspicious of their wheat supply.

138 incidents were reported in 2012 of non-GMO crops being contaminated by GMO crops

There are 158 GM crop events approved in Canada and 181 events approved in the US.