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[Biotechnology, especially genetic modification, and legislation].

Biotechnology and genetic modification (GM) related legislation is not yet fully developed in the European Union (EU). New legislation has been recently issued ('Introduction of GMO's in the environment') and recently proposals from the European Commission ('GMO's in food and feed' and 'Traceability and labelling of GMO's') entered the decision-making process in the end of 2001. The proposals for the establishment of the European Food Authority play a role in this respect. GMO legislation is complex not in the least because of the demands for the dossiers, to be submitted with an application, while these procedures for admission must become more transparent. In this paper the relevant legislation will be discussed with the exception of that related to human health. Because of dissatisfaction with the present legislation, the European Commission in the past years granted no new approvals for introductions on the market of GMO's and for GM novel foods. New legislation should suspend the present de-facto moratorium. The tasks and position of the Inspectorate for the Health Protection and Veterinary Public Health is discussed. A provision has been made in the legislation with respect to adventitious or technically unavoidable contamination of raw materials with GMO's up to a maximum of 1%, of which the enforcement is not yet watertight. The analytical methods are being still developed.

Animal Feed↗

Nutritional properties of tubers of conventionally bred and transgenic lines of potato resistant to necrotic strain of Potato virus Y (PVYN).

The potential effect of genetic modification on nutritional properties of potatoes transformed to improve resistance to a necrotic strain of Potato virus Y was determined in a rat experiment. Autoclaved tubers from four transgenic lines were included to a diet in the amount of 40% and compared with the conventional cv. Irga. The experiment lasted 3 weeks and special attention was paid to nutritional properties of diets, caecal metabolism and serum indices. Genetic modification of potato had no negative effect on the chemical composition and nutritional properties of tubers, ecosystem of the caecum, activity of serum enzymes and non-specific defence mechanism of the rats. Obtained results indicate that transgenic potato with improved resistance to PVY(N): line R1F (truncated gene coding for PVY(N) polymerase in sense orientation), R2P (truncated gene coding for PVY(N) polymerase in antisense orientation), and NTR1.16 (non-translated regions of PVY(N) genome in sense orientation) are substantial and nutritional equivalence to the non-transgenic cultivar. Tubers of transgenic line NTR2.27 (non-translated regions of PVY(N) genome in antisense orientation) increased the bulk of caecal digesta and the production of SCFA as compared to tubers of the conventional cultivar and the other transgenic clones. Taking into account some deviations, it seems reasonable to undertake a long-term feeding study to confirm the nutritional properties of tubers of transgenic lines.

Animals↗

Genetically modified plants for improved trace element nutrition.

Deficiencies of iron and zinc are common worldwide. Various strategies have been used to combat these deficiencies including supplementation, food fortification and modification of food preparation and processing methods. A new possible strategy is to use biotechnology to improve trace element nutrition. Genetic engineering can be used in several ways; the most obvious is to increase the trace element content of staple foods such as cereals and legumes. This may be achieved by introduction of genes that code for trace element-binding proteins, overexpression of storage proteins already present and/or increased expression of proteins that are responsible for trace element uptake into plants. However, even very high levels of expression may not substantially increase the iron and zinc contents unless many atoms of trace elements are bound per protein molecule. Another possibility is to introduce a protein that specifically enhances trace element absorption even in the presence of naturally occurring inhibitors, thus improving bioavailability. Genetically modifying plants so that their contents of inhibitors of trace element absorption such as phytate are substantially reduced is another approach. Increasing the expression of compounds that enhance trace element absorption such as ascorbic acid is also a possibility, although this has received limited attention so far. Iron absorption may be increased by higher ascorbic or citric acid content but require overexpression of enzymes that are involved in the synthetic pathways. Finally, a combination of all of these approaches perhaps complemented with conventional breeding techniques may prove successful.

Absorption↗

Contaminants in feed for food-producing animals.

Outbreaks of Bovine Spongiform Encephalopathy (BSE) and food borne microbial infections, dioxin contaminated animal products, the presence of veterinary drug residues, microbial resistance to antibiotics, mycotoxins, agricultural and industrial chemicals, etc. are serious concerns for the food industry in many countries. Since the direct links between feed safety and safety of foods of animal origin are obvious, feed production and manufacture should be considered as an integral part of the food production chain. Industry is responsible for the quality and safety of food and feed that is produced. This paper is a brief review of some microorganisms as source of infections for farm animals that could result in human illnesses. These include Salmonella enterica, Bacillus anthracis, Toxoplasma gondii, Trichinella spiralis, prions, Listeria monocytogenes, EHEC, Campylobacter, Clostridium botulinum, Hog Cholera virus, Foot and Mouth Disease virus, etc. as well as other contaminants associated with animal feed such as mycotoxins, veterinary drugs, dioxins and PCB and Genetically Modified Organisms.

Animal Feed↗

Biotechnology of temperate fruit trees and grapevines.

Challenges concerning fruit trees and grapevines as long lived woody perennial crops require adapted biotechnological approaches, if solutions are to be found within a reasonable time frame. These challenges are represented by the need for correct identification of genetic resources, with the foreseen use either in conservation or in breeding programmes. Molecular markers provide most accurate information and will be the major solution for questions about plant breeders rights. Providing healthy planting material and rapid detection of newly introduced pathogens by reliable methods involving serological and molecular biological tools will be a future challenge of increases importance, given the fact that plant material travels freely in the entire European Union. But also new breeding goals and transgenic solutions are part of the biotechnological benefits, e.g. resistance against biotic and abiotic stress factors, modified growth habits, modified nutritional properties and altered processing and storage qualities. The successful characterization of transgenic grapevines and stone fruit trees carrying genes of viral origin in different vectors constructed under ecological consideration, will be presented. Beyond technical feasibility, efficiency of resistance, environmental safety and Intellectual Property Rights, also public acceptance needs consideration and has been addressed in a specific project. The molecular determination of internal quality parameters of food can also be addressed by the use of biotechnological tools. Patient independent detection tools for apple allergens have been developed and should allow to compare fruits from different production systems, sites, and genotypes for their content of health threatening compounds.

Animals↗

A general method for selection of alpha-acetolactate decarboxylase-deficient Lactococcus lactis mutants to improve diacetyl formation.

The enzyme acetolactate decarboxylase (Ald) plays a key role in the regulation of the alpha-acetolactate pool in both pyruvate catabolism and the biosynthesis of the branched-chain amino acids, isoleucine, leucine, and valine (ILV). This dual role of Ald, due to allosteric activation by leucine, was used as a strategy for the isolation of Ald-deficient mutants of Lactococcus lactis subsp. lactis biovar diacetylactis. Such mutants can be selected as leucine-resistant mutants in ILV- or IV-prototrophic strains. Most dairy lactococcus strains are auxotrophic for the three amino acids. Therefore, the plasmid pMC004 containing the ilv genes (encoding the enzymes involved in the biosynthesis of IV) of L. lactis NCDO2118 was constructed. Introduction of pMC004 into ILV-auxotrophic dairy strains resulted in an isoleucine-prototrophic phenotype. By plating the strains on a chemically defined medium supplemented with leucine but not valine and isoleucine, spontaneous leucine-resistant mutants were obtained. These mutants were screened by Western blotting with Ald-specific antibodies for the presence of Ald. Selected mutants lacking Ald were subsequently cured of pMC004. Except for a defect in the expression of Ald, the resulting strain, MC010, was identical to the wild-type strain, as shown by Southern blotting and DNA fingerprinting. The mutation resulting in the lack of Ald in MC010 occurred spontaneously, and the strain does not contain foreign DNA; thus, it can be regarded as food grade. Nevertheless, its application in dairy products depends on the regulation of genetically modified organisms. These results establish a strategy to select spontaneous Ald-deficient mutants from transformable L. lactis strains.

Amino Acids, Branched-Chain↗

Genetically modified plants: the stakes.

Generically modified plants (GMP) are massively used on the American continent in Australia and in China, since they represent an unquestionable potential for progress. New attributes are therefore devoted to the human and animal diet, to the facilitating of culture management, to the reducing of the chemical fertilizer and pesticide usage, and to the conquest of new cultural spaces. Considering itself to be flawed by a too hasty plunge into the market, concomitant with sagging evaluations of other innovations, Europe is confronted by a strong societal debate which blocks GMP cultures and orientates the research towards an evaluation of the environmental and public health risks and an evaluation of their economical and sociological impacts. The authors encourage this societal debate in order to arbitrate the presence of transgenes in conventional productions and products, to define the accepted rules of responsibility, to decide what is not acceptable, and to involve the more upstream actors and operators of the innovation process, all that keeping in mind the agronomical, ecological and economical repercussions of their decisions.

Animals↗

[Functional nutrition and optimal nutrition. Near or far?].

The concept of functional food, about which scientific agreement is still lacking, springs from the field of Optimum Nutrition, aimed at modifying genetic and physiological aspects of human life and at the prevention and treatment of a growing number of diseases, far beyond merely covering nutritional requirements. From the European Union perspective, functional foods can be natural as well as industrially processed foods. The leading functional foods regarding which the soundest scientific evidence exists are probiotics, live microbial food ingredients represented mainly by fermented dairy products. Prebiotics, such as inulin-type fructans, are the trophic substrate of probiotics and potential intestinal microflora selectors. The combination of prebiotics and probiotics is termed synbiotic. Innumerable substances are known to have functional effects: soluble and insoluble fiber, phytosterols, phytoestrogens, monounsaturated and polyunsaturated fatty acids, phenol derivatives, vitamins and other phytochemicals. Functional foods exert their actions on different systems, especially the gastrointestinal, cardiovascular and immunological ones, acting too as enhancers of development and differentiation and positively modulating nutrient metabolism, gene expression, oxidative stress and the psychic sphere. The establishment of Health Claims must be firmly based upon scientific knowledge and legal regulation. Efficient biomarkers related to biological response must be found. Furthermore, it is essential to analyze possible diet or drug interactions as well as it is indispensable to conduct valid studies on humans. The prime objective must be the diet as a whole. Thus, the future challenge of a functional diet emerges.

Dietary Fiber↗

Planktonic/sessile dimorphism of polysaccharide-encapsulated sphingomonads.

Sphingomonads have acquired diverse metabolic activities to inhabit a wide range of environments. Several strains of Sphingomonas display phenotypic dimorphism and can adopt either a planktonic or sessile behavior in liquid media. The sessile state is marked by the presence of a viscous exopolysaccharide capsule. Specific types of these capsular polysaccharides are harvested from large-scale fermentations for use as rheology modifiers in many industrial and food applications. Sensing of environmental stimuli and genetic control over synthesis of the capsule are key events in alternating between these two phenotypes.

Journal Article↗

The fate of the recombinant DNA in corn during composting.

In order to make regulations that safeguard food and the environment, an understanding of the fate oftransgenes from genetically modified (GM) plants is of crucial importance. A compost experiment including mature transgenic corn plants and seeds of event Bt 176 (Zea mays L.) was conducted to trace the fate of the transgene cryIA(b) during the period of composting. In bin 1, shredded corn plants including seeds were composted above a layer of cow manure and samples from the corn layer were collected at intervals during a 12-month period. The samples were tested for the transgene persistence and microbial counts and also the compost was monitored for temperature. In bin 2, piles of corn seeds, surrounded by sheep manure and straw, were composted for 12 months. A method combining nested polymerase chain reaction (PCR) and southern hybridization was developed for detection of the transgene in compost. The detection sensitivity was 200 copies of the transgene per gram of dry composted corn material. Composting commenced on day 0, and the transgene was detected in specimens from bin 1 on days 0 and 7 but not on day 14 or thereafter. The transgene in corn seeds was not detectable after 12 months of composting in bin 2. Temperatures in both bins rose to about 50 degrees C within 2 weeks and remained above that temperature for about 3 months, even when the ambient temperature dropped below -20 degrees C. Extracts from compost were inoculated onto culture plates and then were incubated at 23 to 55 degrees C. Within the first 2 weeks of composting in bin 1, the counts of bacteria incubated at 55 degrees C increased from 3.5 to 7.5 log10, whereas those incubated at 23 degrees C remained at about 7.5 log10. The counts of fungi incubated at 45 degrees C increased slightly from 2.5 to 3.1 log10, but those incubated at 23 degrees C decreased from 6.3 to 3.0 log10. The rapid degradation of the transgene during composting of Bt corn plants suggested that the composting process could be used for safe disposal of transgenic plant wastes.

Animals↗

Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods.

This article summarizes the current situation pertaining to modern biotechnology in Ireland, with a particular focus on genetically modified (GM) crops. It briefly examines some important results of the major national surveys carried out in Ireland since 1989, highlights the recent upsurge in media (newspaper) coverage of GM related stories in three Irish opinion leader publications and it allows for an insight into the Irish public's relationship with modern biotechnology.

Attitude↗

[Transgenic plants: towards a more balanced diet?].

Over the past ten years, knowledge of the metabolic capacities of plants has expanded considerably. Combined with technical achievements in gene transfer, a better understanding of metabolic regulation provides new more precise and targeted means of modifying plant products in order to improve health and well being through diet. While a great number of attempts to modify plant metabolism are still in the exploratory phase, some key applications are emerging with applications involving micro and macronutrients.

Amino Acids↗

Model studies on the detectability of genetically modified feeds in milk.

Detecting the use of genetically modified feeds in milk has become important, because the voluntary labeling of milk and dairy products as "GMO free" or as "organically grown" prohibits the employment of genetically modified organisms (GMOs). The aim of this work was to investigate whether a DNA transfer from foodstuffs like soya and maize was analytically detectable in cow's milk after digestion and transportation via the bloodstream of dairy cows and, thus, whether milk could report for the employment of transgene feeds. Blood, milk, urine, and feces of dairy cows were examined, and foreign DNA was detected by polymerase chain reaction by specifically amplifying a 226-bp fragment of the maize invertase gene and a 118-bp fragment of the soya lectin gene. An intravenous application of purified plant DNA showed a fast elimination of marker DNA in blood or its reduction below the detection limit. With feeding experiments, it could be demonstrated that a specific DNA transfer from feeds into milk was not detectable. Therefore, foreign DNA in milk cannot serve as an indicator for the employment of transgene feeds unless milk is directly contaminated with feed components or airborne feed particles.

Animal Feed↗

[Transgenesis; applications, welfare and ethics].

Biomedical proteins and proteins having a nutritional function can be produced in large quantities (as well as cheap) in the mammary gland of mammals such as cows. Several of these proteins are of major economic importance and will result in several new uses for milk. For example, milk of genetically modified cows will be produced especially for branches of the pharmaceutical industry or for the baby food industry. Production of these new proteins will not have any injurious effects on the animal as proteins in the mammary gland are characterised by the fact that they follow a one-way course.

Animals↗

Interlaboratory transfer of a PCR multiplex method for simultaneous detection of four genetically modified maize lines: Bt11, MON810, T25, and GA21.

The number of cultured hectares and commercialized genetically modified organisms (GMOs) has increased exponentially in the past 9 years. Governments in many countries have established a policy of labeling all food and feed containing or produced by GMOs. Consequently, versatile, laboratory-transferable GMO detection methods are in increasing demand. Here, we describe a qualitative PCR-based multiplex method for simultaneous detection and identification of four genetically modified maize lines: Bt11, MON810, T25, and GA21. The described system is based on the use of five primers directed to specific sequences in these insertion events. Primers were used in a single optimized multiplex PCR reaction, and sequences of the amplified fragments are reported. The assay allows amplification of the MON810 event from the 35S promoter to the hsp intron yielding a 468 bp amplicon. Amplification of the Bt11 and T25 events from the 35S promoter to the PAT gene yielded two different amplicons of 280 and 177 bp, respectively, whereas amplification of the 5' flanking region of the GA21 gave rise to an amplicon of 72 bp. These fragments are clearly distinguishable in agarose gels and have been reproduced successfully in a different laboratory. Hence, the proposed method comprises a rapid, simple, reliable, and sensitive (down to 0.05%) PCR-based assay, suitable for detection of these four GM maize lines in a single reaction.

Base Sequence↗