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ADDIS ABABA UNIVERSITY COLLEGE OF HEALTH SCIENCES SCHOOL OF VETERINARY MEDICINE Term Paper Submitted for the Course

Biotechnology (ANPS-802)

TERM PAPER ON: Biotechnology in Animal Feed: Improvement of Feeds and Rumen Organisms

By: DEHINENET GEZIE

JUNE, 2011 DEBRE ZEIT, ETHIOPIA

TABLE OF CONTENTS

TABLE OF CONTENTS ............................................................................................................................. I ACKNOWLEDGMENTS ......................................................................................................................... II ABBREVIATIONS ................................................................................................................................... III SUMMARY ............................................................................................................................................... IV 1. INTRODUCTION ....................................................................................................................................1 2. DEFINITION OF BIOTECHNOLOGY ................................................................................................3 3. CONTRIBUTION OF BIOTECHNOLOGY TO LIVESTOCK REVOLUTION.............................4 4. TECHNOLOGICAL OPTIONS .............................................................................................................5 4.1. Plant Breeding and Plant Biotechnology............................................... Error! Bookmark not defined. 4.1.1. Agronomic Improvements ............................................................. Error! Bookmark not defined. 4.1.2. Nutritional Improvements Value Added Feed Stuff ................... Error! Bookmark not defined. 4.2. Animal and Microbial Biotechnology .................................................... Error! Bookmark not defined. 4.3. Biological Treatment or Microbiological Process ................................ Error! Bookmark not defined. 4.3.1. Microbial Biomass Protein (MBP) or Single Cell Protein Production .........Error! Bookmark not defined. 4.3.2. Lignin Biodegradation (Whole Substrate Feeding) ....................... Error! Bookmark not defined. 4.3.3. Mushroom Production and Utilization of Spent Straws as Cattle Feed ........Error! Bookmark not defined. 4.4. Silage Inoculants (For Succulent Roughages)....................................... Error! Bookmark not defined. 4.5. Treatment to Increase Digestibility ....................................................... Error! Bookmark not defined. 4.5.1. Improving the Digestibility of Crop Residues by Chemical Treatments ......Error! Bookmark not defined. 4.5.2. Microbial Treatment of Straw and Other Crop Residues to Improve Digestibility ............... Error! Bookmark not defined. 4.6. For Concentrate Feeds ............................................................................ Error! Bookmark not defined. 4.6.1. Antibiotics or Antimicrobial Agents .............................................. Error! Bookmark not defined. 4.6.2. Feed additives ................................................................................ Error! Bookmark not defined.

6. CONCLUSION .......................................................................................................................................19 REFERENCES ...........................................................................................................................................25

ACKNOWLEDGMENTS

I would like to acknowledge Dr.Tesfay, S. for giving me a chance to prepare this term paper. My acknowledgment also goes to the authors who are cited in this review for their valuable ideas which is incorporated in.

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ABBREVIATIONS AI bST CFU ET GM MBP MIT MOS NDRI STS UMMB USDA SMS artificial insemination bovine somatotropin colony forming units embryo transfer genetically modified Microbial Biomass Protein Massachusetts Institute of Technology Mannan-oligosaccharides National Dairy Research Institute somatotropins molasses mineral block United State Department of Agriculture Spent Mushroom Substrate

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SUMMARY

It is expected that in the coming years increased urbanization and rising incomes will lead to substantial growth in the demand for chicken meat, eggs and milk. Therefore livestock industry would need to utilize biotechnological inputs like genetically modified feedstuffs and feed additives in order to improve nutrient balance for maximum feed utilization. Amongst the feed additives, enzymes would play a major role where the usage will be selective and specific for various ingredients. This will allow nutritionists to formulate feed with alternate feed resources. Ingredients like corn and Soya which are considered to be the most ideal ingredients could be further enhanced nutritionally by genetic modification for complete nutrient digestibility. Due to intensified farming the industry will need to take advantage of modern biotechnology to decrease problems of wet litter, odor, excess of phosphorus and nitrogen in animal waste. Other biotechnologically derived feed additives like amino acids, probiotics, prebiotics, organic minerals, herbal performance enhancers, toxin binders etc will also be used in a significant manner to achieve maximum productivity.

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1. INTRODUCTION

The demand for food of animal origin is increasing in the world due to economic growth, population growth and urbanization, which in turn means greater use of cereals and oilseeds for animal feed. This is going to be the trend in most developing countries (Delgado et al., 1999). In some decades the increase in demand were coped mainly by expanding livestock population. However, declining land areas per agricultural population forces to intensify livestock production. The emphasis so far has been on disease management, labor efficiency, management practices and increasing yields. Today, we are faced with an extraordinary set of challenges of increasing food production of animal origin with all the other limitations like land, water, weather etc and the question is how would we meet these demands. We also have another challenge that the food we produce has to be highly cost efficient to make it more sustainable. This means that the real prices of milk, eggs, meat and fish would be falling. In such a situation biotechnology would potentially play a very significant role in animal husbandry, covering the entire food production chain from strain development, diagnostics, preventive therapies, nutritive improvements to product processing. The biotech revolution has already reached in different countries and various organizations are actively engaged in research and development activities to produce biotechnological products for the improvement of productivity and productive efficiency. The focus of this paper, therefore, is to discuss the present controversial ideas of animal biotechnology.

Key words: Animal feed, biotechnology, microbial treatment E. Sachez, M. Ragucci And A. Vassarotti (2002): Ethical, Legal and Socio Economic Aspects of Agriculture, Fisheries and Food Biotechnology, Belgium,P,180.

Introduction

In the debate about the ethical aspects of biotechnology, most, if not all attention has been focused on ethical thinking in society mainly from the perspective of public acceptance of biotechnology and on the role of ethical arguments in this respect. The ethical opinions in general in society and more specific of consumer groups on biotechnology and biotechnological products and the differences in national cultures in relation to public acceptation, have been the subject of a number of national studies (E. Sachez, 2002).

The use of GMOs in agriculture and food has become one of the most controversial topics in contemporary societies. Promoters of agricultural biotechnologies are concerned that the current public controversy on GMOs is impeding the development and commercialisation of a new technological field considered to be of strategic economic importance. At the same time, critics who believe that GMOs involve unacceptable impacts on the environment, health and society, continue to feel that their concerns have not been addressed. The public is somehow caught in the middle. What do people in Europe think about the use of genetically modified organisms in agriculture and food? What expectations and concerns do they have? How do they shape their views when faced with a new issue such as this? How do they perceive this issue within the whole context of modernisation and lifestyle changes? (E. Sachez, 2002).

During the last few years agricultural biotechnologies have been the subject of numerous inquiries, consultation exercises and public debates and the number continues to grow yet most protagonists, on both sides, remain dissatisfied. The need to understand public responses to biotechnology has never been more pressing. But understanding the response of policy makers to perceived public concerns is also essential (E. Sachez, 2002). The focus of this paper, therefore, is to discuss the present controversial ideas of animal biotechnology.

2. DEFINITION OF BIOTECHNOLOGY Biotechnology is defined as the application of technologies, such as recombinant DNA techniques, biochemistry, molecular and cell biology, genetics and genetic engineering, and cell fusion techniques etc or using living organisms or its products, to manufacture industrial products including antibiotics, insulin, and interferon, to improve plants or animals, to develop microorganisms for specific uses, to identify targets for pharmaceutical development, to transform biological systems into useful processes and products or to develop organisms for specific uses.

3. CONTRIBUTION OF BIOTECHNOLOGY TO LIVESTOCK REVOLUTION

The present status of poultry, cattle and aqua industries pose challenges and opportunities for improvement of productions further to meet the growing demands in the coming years. So far chemical, biological and mechanical innovations had come to our rescue to increase productivity but now the most promising technology would be Biotechnology. Biotechnology is a modern tool by which crop and animal improvement is possible by applying cell and gene technology. Agricultural biotechnology brings together advanced disciplines of biology, genetics, molecular biology, biophysics, biochemical engineering and computer science. The potential contribution of Biotechnology to the livestock revolution would be in area of reproductive biotechnology which includes artificial insemination (AI) and embryo transfer (ET) and molecular biotechnology which consists of development of transgenic animals, diagnostic kits, recombinant vaccines and genetically modified (GM) feed stuff and feed additives. In the production of transgenic animals especially for poultry and cattle, a different country like India is unlikely to make great progress in the near future. This is because the technology development is very time consuming, expensive and inefficient at present. Also consumer acceptance, ethical considerations and risk evaluation may reduce the scope considerably. But other biotechnological inputs covering the entire food production chain from animal feed to product processing would be applicable. The largest impact of biotechnology on livestock production is likely to stem from increasing the livestock feeds through improving nutrient content as well as the digestibility of low quality feeds through use of efficient feed additives.

4. ANIMAL BIOTECHNOLOGIES` BENEFITS AND CONCERNS I. REPRODUCTIVE TECHNOLOGIES

1.Artificial insemination Various aspects of artificial insemination technology have been fairly standardized. It has now become a practical technology in commercial dairy cattle programs in both developed and developing countries, covering a worldwide total of about 50 million first inseminations (Chupin and Schuh 1993, Chupin and Thibier 1995). In India, where well structured dairy development programmes have been established with cattle and buffalo, artificial insemination (AI) procedure has played a valuable role in facilitating appropriate genetic change in animal populations, being linked to intensive male selection. In developed countries, AI system is increasingly being developed and used in the breed development of sheep, horses and pig genetic resources. AI technology offers certain specific advantages, especially i.widespread use of outstanding males and dissemination of superior genetic material; ii.progeny testing under environment and managerial conditions to improve the rate and efficiency of genetic selection; iii.accelerated introduction of new genetic material by import of semen rather than live animals and thus, reducing the international transport costs; iv.enabling the use of frozen semen even after the donor is dead; and v.reducing the risk of spreading sexually transmitted diseases. Long-term semen storage, without loss of viability, for use in AI, is another valuable technology for promoting conservation of endangered breeds of farm animal species, although this technology has the disadvantage of preserving only half genotypes and requiring secure cryopreservation facilities. 2.Embryo Transfer Considerable progress has been made in methods of recovery, storing and implanting cattle embryos (including drug applications to enable acceptance of implanted embryos) in several counties of the world. Approximately 460,000 bovine embryos were transferred world- wide in 5

1997 (Thibier 1998). Several Multiple Ovulation Embryo Transfer schemes (MOET) have been initiated and about 10% genetic gain has been achieved in cows (Nicholas and Smith 1983, Lohius 1995). In India also, techniques and protocols of ET technology are being standardized in cattle, buffalo, sheep, goat, camel, and other species of animals ( Madan et al. 1993, Misra 1993, Anon 1998-99). Embryo transfer and other associated reproductive technologies have been successively used for rapidly multiplying the populations of elite breeds of cattle (Willet et al., l951, Thibier 1996), buffalo (Drost et al. 1983, Madan et al.1993, Misra 1993, Misra et al 1999), sheep (Holm et al. 1996, Thibier 1996), goat (Armstrong et al. 1983, Pawshe et al. 1994, Thibier 1996), horse (Riera and McDonough 1993, Squires et al. 1999) and pig (Kvansnickii 1951, Galvin et al. 1994, Hazeleger and Kemp 1999). ET/MOET schemes are especially recommended for some specialized purposes, viz., (i) rapid expansion of rare and/or improved exotic/local genetic stocks of farm animals and even elite female animals (ii) reducing the cost of international transport by shipping embryos rather than living animals in which case quarantine restrictions would also apply; (iii) the rapid replacement of existing genotypes by using ET/MOET rather than grading up through repeated crossing; and (iv) the possibility of increasing the twinning rate by combining AI with the transplanted embryo (Cunningham 1999). Other advantages of ET technology are (i) pre-natal sex determination (ii) control of sexually transmitted diseases (iii) using biological diversity of embryos and (iv) environmental adaptability as compared to transport and establishment of imported animals. In combination with other animal biotechnological procedures, ET/MOET techniques can accelerate herd development and animal conservation especially of rare genetic stocks. Embryo splitting technique and transfer of split embryos have been standardized by National Institute of Immunology (NII), and the first bovine split embryo derived calf was born in November, 1988 (Raghupathy and Hasnain 1991; Ramachandran 1991. Such technologies enabled the production of 150 cows from an elite cow, as opposed to ten cows that she would have normally produced in her lifetime in conventional husbandry. Scientists at NII also succeeded in producing a 100% pure HolsteinFriesian calf, using a local stray cow as surrogate mother (Raghupathy and Hasnain 1991, Ramachandran 1991). 6

3.Embryo cryopreservation Freezing of semen and embryo is an established commercial practice especially in cattle. About 800 experimental reports and papers have been published on various aspects of embryo cryobiology, especially on developing effective methods of preserving viability by freezing and quick thawing in the presence of cryoprotective additives (Leibo and Loskutoft 1993). This technique is useful as a conservation strategy for endangered breeds, and every effort should be made to select embryos representing the maximum range of current diversity. Preservation of embryonic stem cells could represent an important method of genome conservation. 4.IVM/IVF Embryo Production Although Brackett et al. (1982) used in vivo matured oocytes for IVF system to produce the first calf in the world, it is now possible to adopt a fully in vitro system, whereby the immature oocytes are matured in vitro, followed by fertilization with in vitro capacitated spermatozoa and the newly formed zygotes are cultured in suitable media for development up to the transferable stage. The birth of such in vitro animals was reported in cattle (Gordon and Lu 1990), goat (Hanada 1985), pig (Cheng et al. 1986) and buffalo (Madan et at. 1994). Despite of several advantages of in vitro embryo production, initial application in both cattle and buffaloes has been limited by the ability to recover oocytes. However, recent development of low invasive ultra sound guided transvaginal oocyte retrieval (TVOR)/ oocyte pick up (OPU) has removed these difficulties to a large extent (Pieterse et al. 1991). This repeated recovery permits production of more embryos than might be possible by standard ET practice (Kruip 1994). TVOR also allows repeated collection of oocytes from endangered species of livestock or livestock of high economic importance in order to propagate such genetic resources in much faster way. Gamete intra-fallopian tube Transfer (GIFT), an assisted reproductive technology has been used to produce pregnancies in mares (Carnevale 1996, Hinrichs et al. 1998 a, b). Intra-cytoplasmic sperm injection (ICSI), another assisted reproductive technology, has been successfully used in rabbits (Hosoi et al. 1988), cattle (Goto et al. 1991), mares (Squires et al. 1996, Cochran et al. 1998), dogs (Foul ton et al. 1998), pig (Kim et al. 1998), horse/ zebra (Li et al. 1994, Meintjes et al. 1995). 7

Current status of research on IVM/IVF embryo production and cryopreservation of such zygotes was reviewed by Tervit (1997) and Pieterse et al. (1988). This technique is used for bulk production of high quality embryos. Freezing of high quality oocytes and semen would provide germplasm for undertaking important future mating regimes flexibility in the conservation programmes. 5.Sexing semen and embryos It is possible now to extract one cell from an early embryonic stage and with the use of a DNA probe, one can know the sex of the embryo (Thibier and Nibart 1995). Another way is the sorting of semen, one sperm at a time, into males and females, using staining procedure and detecting by laser beam with the help of standard flow cytometry equipment (Johnson et al. 1996). Thus, sperms with an X or Y chromosome could be used to produce male or female embryos /animals. Seidel et al. (1997) suggested procedures for using low sperm count, though with reduced conception rate in heifers. The bovine Y- chromosome specific sequences are conserved amongst buffalo, Indian zebu and Taurus cattle (Appa Rao et al. 1993). Thus, the use of bovine Y-chromosome specific primers, demonstrate the sex of buffalo or Indian zebu cattle embryos. Efficient embryo biopsy method has also been developed (Taneja et al. 1998). The high rate of survival and conception rates of biopsied embryos also clearly reflects the minimal damage of the embryos. Buffalo Ychromosome specific probe was also developed recently (Appa Rao and Totey 1999). 6.Cloning The first successful cloning in domesticated animals was achieved, using early embryonic source material (in sheep, Willadsen 1986, Campbell et al. 1996; cattle, Sims and First 1993, Stice et al. 1996; pig, Prather et al. 1989), nuclear transplantation (Wolfe and Kraemer 1992), embryonic cultured cell line (Wheeler et al. 1994, Campbell et al. 1996) and somatic cells of adult animals ( Dolly sheep Wilmut et al. 1997, cattle, Cibelli et al. 1998). Eight cloned calves were produced from cumulus and oviduct epithelial cells of an adult cow (Kato et al. 1998). Cloning using somatic cells offers opportunities to select and multiply animals of specific merits (Cunningham 1999). Cloning holds the promise of bypassing conventional breeding procedures to allow 8

creation of thousands of precise duplicates of genetically engineered animals or other animals in a single generation. In remote areas, where sampling and storage of adequate samples of semen and embryos is not practical, one could use clonal samples from diverse animals for conservation of the available genetic diversity of such threatened genetic resources (Anon 1998, Cunningham 1999). Due to increasing commercial pressure, several indigenous breeds of cattle (Rathi, Tharparkar, Red Sindhi and Sahiwal etc.) are under threat from imported breeds that are being used in intensive farming systems. The local breeds may contain valuable genes that confer adaptation, especially to heart tolerance or disease resistance, and there is an urgent need to prevent their extinction. Current method of genetic conservation involves storage of frozen semen or embryos but this procedure is costly and time consuming. Cloning may eventually prove to be much simpler and effective means of conservation of breeds. Skin biopsies, hair follicles, and blood samples might be suitable sources of cells, which could be grown briefly in the laboratory, and subsequently frozen in liquid nitrogen for long- term storage. Many animal species are in danger of extinction. In India, one horned rhinoceros, swamp deer, Manipur bro antler deer, hispid hare, wild buffalo, Assam root turtle, pigmy hog, and Bengal florican, are classified as highly threatened animals in their present habitats. Cloning technique might be applied to increase the population sizes of endogenous species or even restore them following extinction. A broad spectrum of biodiversity can be collected and cryopreserved at modest cost (Benford 1992). San Diego zoos center for reproduction of endangered species has created a collection of frozen fibroblast cell cultures and tissue samples collected from various rare and threatened animal species, such as, Prezewalskis horse, Sumatran rhinoceros etc. China has announced the initiation of a programme to increase the giant panda populations, using nuclear transfer techniques involving even the possible use of bear as egg donors and surrogate mothers (Corley-Smith and Brandhorst 1999). In cross-breeding programmes involving Bos taurus dairy cattle breeds and Bos indicus local breeds in tropical countries, heterosis unto 25% has been regularly achieved in the first cross (Cunningham and Syrstad 1987). Since subsequent breeding strategies normally do not retain the achieved heterosis, embryos could be produced by IVF, using Bos indicus semen on Bos taurus 9

oocytes collected in vivo or alternatively one could use embryos cloned from cells of high performing F1, individuals (Jarvis 1996). Principal germ cell technique is now being developed for producing germ line chimeras and transgenic poultry (Samkiss 1997, Cunningham 1999). Isolation of embryo-derived cell lines has been reported from preimplantation embryos of mouse, Syrian Golden hamster, rat, mink, pig, cattle and sheep (see review by Wheeler et al. 1994). II.GENETIC TECHNOLOGIES 1.Genome maps Recent progress in genetic linkage mapping within multiple animal species has incorporated the genomes of cattle (Bishop et al. 1994, Barendse et al. 1994), swine (Rohrer et al. 1994), sheep (Crawford et al. 1994), and poultry (Khatib et al. 1993). At present the total number of mapped loci are 2850 for cattle, 1774 for pigs and over 1000 for sheep, while the highest resolution single linkage maps in each species are 1445 for cattle, 1250 for pigs and 500 for sheep (Kappes et al. 1999). Technological break through has provided an opportunity to simultaneous construct genetic linkage and physical maps for the major livestock species and merge them into comprehensive maps (Beattie and Hruska 1994). These include linkage PCR (Polymerase chain reaction) amplified highly polymorphic, repetitive elements (micro satellites, Weber and May 1989), and direct chromosomal assignments of these polymorphic elements either by direct amplification (Troyer et al. 1994a / 1994b) or through the use of larger pieces of DNA such as cosmids and florescence in situ hybridization (Yerle et al. 1994, Hawkin et al. 1994). Construction of livestock genome maps has also been benefited by simultaneous development of the data- bases capable of integrating genetic linkages and physical and comparative mapping data (Beattie and Hruska 1994). Furthermore, the co-ordination by FAO within the global strategy of the procedural aspects of genetic resources sampling, analytic methodologies and result reporting of the genetic distancing research activities, which are getting underway for each animal species, could help involve developing countries, and rapidly realize cost-effective country and regional conservation decision making for the large number of animal genetic resources currently identified as at risk (Cunningham 1999). The rapid multiplication of superior germplasm can be attained through the techniques of embryo transfer coupled with in vitro fertilization, twinning, and embryo splitting, 10

sexing and cloning. Several National Institutes viz., Indian Veterinary Research Institute, National Dairy Research Institute, National Institute of Immunology and National Diary Development Board, have already initiated research and development programmes using these biotechnologies. No assessment of impact of these technologies on our cattle and buffalo populations has been made so far. 2.Transgenesis Since the report of first transgenic animals (mouse, Gordon and Ruddle 1981;sheep and pig, Hammer et al., 1985; sheep, Simon et al. 1998; goat, Ebert et al. 1991;and cattle, Krimpenfort et al 1991, Cibelli et at. 1998), engineered gene constructs have also been transferred to birds and fish. Several biotechnological techniques viz., pro-nuclear micro-injection (farm animals), cytoplasmic micro-injection (fish), retrovirus based vectors (birds), are presently used to produce transgenic animals. The present technology allows survival rate of one in ten injected and transferred embryos, and about one in ten of these carries the Tran gene (Wall 1996). Gene transfer in animals has been aimed at (i) modifying the fat or protein synthesis in the mammary glands (Vilotte et al. 1997), (ii) transferring of growth hormone gene in pigs (Pursel and Rexroad, 1993), (iii) transferring cysteine synthesis gene into sheep for enhancing wool production (Word and Nanerrow 1991, Powell et al. 1994), (iv) transferring cold tolerance gene from flounder into salmon (Hew et al. 1995) and (v) micro-injecting gene imparting resistance to influenza virus in pig (Muller et al 1991, 1992). Another king of target protein could be the one, which is not a part of normal animal functioning of a specific species but for which this species could be an economic medium for production. Transgenic bacteria are now routinely producing human insulin. Similarly, although methods are now available to produce recombinant proteins in several species of micro- organisms, plants or animals, the success depends on economics of production, the maximum success so 1 anti-trypsin isfar, being in using milking animals (Bremul 1996). The human being expressed in sheeps milk at levels high enough for commercial extraction (Wright et al. 1991). High level expression of several transgenic encoding medically important proteins have been accomplished in the transgenic goat and most notably a-1 antitrypsin at 15 -30 g / lit in sheep (carver et al.1993), a11

lactalbumin at 4 -5 g / lit in cow (Colmon 1996), acid alpha-glucosidase at 1-2 g / lit in rabbit (Hersbach 1997), and tissue plasminogene activator at 6 g / lit in goat and antithrombin III at 10 g / lit in goat (Ziomek 1988). The potential for the synthesis of pharmaceutical products in milk has continued to be exploited, and seven such products are presently being produced with the estimated market value of US $ 3 billion (Wall 1996). The feasibility of developing transgenic vaccines for major diseases of farm animals, especially for diseases caused by viruses which cannot be grown in cell cultures, has also been explored, some of which seem to have potential for use in resource poor countries. (Cunningham 1999). Although animal biotechnology has revolutionized the generation of veterinary biologicals and has improved significantly the livestock health by providing cheaper and effective vaccines with long shelf life and not requiring elaborate cord chain, care should, however, be exercised in developing new biologicals so that they do not pose any threat to the safety of environment by safeguarding the overall health and well being of plant, animal and human life (Natrajan and Rasool 1997). The diagnostic uses of modern animal biotechnology are other areas of R &D efforts, especially, the use of recombinant proteins and gene deletion mutant vaccines for use in disease diagnosis (Cunningham 1990). Recombinant proteins and monoclonals provide means to diagnose diseases with specificity, accuracy, rapidity, uniformity, manipulatability and ease of performance, while nucleic acid hybridization detects early infection (Natrajan and Rasool 1997). 3.Gene identification The increasing knowledge of mammalian genetic structure has helped in developing DNA based traceability for meat industry, especially to provide widespread consumer guarantees of traceability of products. (Meghen et al. 1998) 4.Molecules conservation 12

Genetic characterization of populations of target species, using a range of micro-satellite loci, can reveal a great deal about the evolutionary history of population and of their relationships with other populations (Mac Huge et al. 1997). Variations in maternal mitochondrial and paternal Y chromosome can provide additional criteria for analyzing patterns of gene flow between populations (Bradley et al. 1998). 5.Germplasm conservation One of the prerequisites of effecting animal genetic resources conservation is the identification of populations to be conserved. Several techniques viz., Restriction fragment length polymorphism (RFLP), random amplification of polymorphic DNA (RAPD), single strand conformation polymorphism (SSCP), mini satellites and micro satellites etc., are used for such a purpose. An estimation of average heterozygocity of a population helps in deciding if it should be preferred for preservation over other populations. Populations with maximum homozygocity need to be preserved because those have the least potential survival value. The above said techniques help in determining the homozygocity / heterozygocity at several loci simultaneously, thereby assisting to provide an index of heterozygocity. Embryo banks, sperm banks, egg banks and DNA banks serve as an insurance against any eventual erosion of genetic diversity of animal species, and their conservation for posterity. Coupled with cryopreservation, invitro fertilization and other reproductive technologies, discussed above, permit medium to long-term storage of genetic material and restoration of eroded genetic diversity especially when in situ conservation is not possible. DNA marker and other molecular conservation techniques assist in animal breed cataloguing. Under the WTO era, every country is conscious about creating clear-cut identities of its own animal genetic resources. DNA techniques, using pooled DNA samples from random individuals, can generate breed specific DNA signatures, which can be developed as identification marks. The FAO Global Strategy for the Management of Farm Animal Genetic Resource places strong emphasis on the use of molecular methods to assist the conservation of endangered breeds and the Measurement of Domestic Animal Diversity aimed at reliably establishing the pair wise genetic distances amongst breeds of target species (Barker et al. 1993). An informal group of 13

experts of FAO is identifying the common set of micro-satellites, which should be used globally for each farm animal species (Bradley et al. 1996). III.RECOMBINANT HORMONES AND IMMUNO-MODULATION 1.Bovine Somatotropin Bovine somatotropin (rBST), a natural growth hormone secreted by the anterior pituitary in all animals, has a major effect on the regulation of growth. It is now possible to transfer the DNA sequences responsible for somatoropin synthesis into bacteria and to produce large quantities of somatoropin commercially at economic levels. Milk production increased between 10 (5 mg BST/day) to 20% (20 mg BST/day) using genetically engineered rBST (Chilliard 1989, Bauman et al. 1994). In view of public concern about the safety of its use in dairy cows, exhaustive evaluation tests were conducted in USA and its use was found safe and permitted for commercial use (Juskevich and Guyer 1990). High production makes higher demands on animal physiology and if adequate feed supply is lacking, negative effects are observed on fertility besides other health problems especially mastitis and ketosis, and the increased frequency of twins (Phipps 1989). In India no serious attempt has been made so far to produce recombinant somatotropin, and thereby to increase milk production in commercial establishments. One of the problems associated with this approach is the high intake of feed, which lowers the inputoutput ratios, thereby reducing the profit margin. 2.Immunomodulation Immunomodulation experiments aimed at modifying endogenous hormones function have shown to improve fertility, growth, lactation and body composition in farm animals (Meloen 1995, Terqui et al. 1995). Active immunization is preferred for long-term responses such as immunocastration, while passive immunization is more appropriate for eliciting short term responses, viz., increased milk yield (Pell and Aston 1995). An animal birth control injection, called Talsur was developed by scientists at the NII, for sterilization of male animals. The injection is usable in all mammals, but would be particularly 14

useful for sterilization of bulls to stop the proliferation of animals of low economic value. Furthermore, as the bulls retain libido, they could be used as teasers bulls to identify females in oestrus. Insemination with semen of high genetic stocks at appropriate time enhanced the chances of conception, as the fertile life of the egg was only 1-2 days. Tulsur has also found application in the sterilization of stray dogs (Rajhupathy and Hasnain 1991). Increases in growth hormones (GH) are known to increase the efficiency of feed use for protein synthesis, but limited experiments conducted on rodents, sheep, goats and cattle have so far not produced consistent results (Flint 1995). Interactions between hormonal and immune system are very complex, nevertheless several successful products e.g. fecundin, vexstrate and others, have been marked for enhancing animal productivity, mostly under specialized production regimes (Cunningham 1999). 3.FEED ENHANCEMENT Biotechnological contributions to livestock nutrition include single cell protein production, the genetic modification of nutritive value of forages, probiotic and biotic feed additives, and the use of enzymes for enhancing the nutritive value and quality of feeds (Cunningham 1999). Cellulose and hemi-cellulose enzymes, as silage additions, as well as bacterial inoculates for preservation and improvement in digestibility, are also available besides other possibilities for feed enhancement (Robinson and McEvoy 1993). These technologies assist in the effective utilization of low nutrient content feedstuff into animal products. The current approach is to survey the naturally available organisms and select the best ones. Productivity in ruminants can be enhanced by using other biotechnological approaches. Thus, not only genes from rumen microorganisms have been cloned mostly in E. coli, but also foreign genes (mostly imparting antibiotic resistance) have been transferred into and expressed in rumen bacteria (Wallace 1997). Gregg (1995) succeeded in transferring a detoxifying (dehalogenase) gene from the soil bacterium, Morexella species into the rumen bacterium, Butyrivibrio fibrisolvens. The modified organism was stable and functional in both the culture medium and in the sheep rumen.

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Another biotechnological innovation enhances animal productivity with the administration of ionophores, which disrupt the functioning of the bacterial membrane and selectively suppress gram-positive bacteria, resulting in shifting the volatile fatty acid production and improvement in feed conversion efficiency (Cunningham 1999). IV.CONCERNS 1.Issues Animal biotechnological innovations and their beneficial effects, discussed in this presentation, fall into four main categories, viz., reproductive, genetic, recombinant hormones, immunomodulation and feed enhancement. Of these innovations and related issues, the most important and contentious ones are (i) the regulation of the testing and utilization of transgenic animals in agricultural environments, (ii) immunology and (iii) the trans-species transmission of diseases. As compared to transgenic plants, there has been limited application of transgenics in animal production environment. Because of the contained nature of most present uses of transgenic animals, with the lesser risk of unwanted escapes into the environment, there has not been much concern in public mind on this issue. There has been, however, a public controversy over the approval and adoption of recombinant bovine Somatotropin (rBST). The main ethical concerns are (i) the social consequences associated with the restructuring of the dairy industry (Kalter 1985, Hallberg 1992); (ii) environmental impacts of dairy restructuring (Lanyon and Beegle 1989); (iii) impact on animal health and well being (Comstock, 1988), and (iv) integrity of the food industry, the regulatory process, and uncertainties about the safety of rBST milk expressed by different research organizations (Thompson 1992, 1994). Besides these considerations, there are differences based on religious believes related to the extent to which scientists may interfere with animals per se and animal genomes (for details on these and other issues, see Megloughlin 1994, Nilsson 1997). There has been little concern in public about the applications of transgenic animals for pharmaceutical purposes and in bio-medical research. However, the world debate has centered around several issues (Weil 1996) viz., public views on (i) humankind and its relationship with nature, especially commodification of nature by humans and antagonism between agrarian societies vs. industrialization; (ii) adverse social and economical impacts, especially benefits to corporations vs. small farmers; (iii) biosafety (viz., harmful side effects in recipient animals and humans) and environmental risks (especially unpredictable expressions in 16

an alien environment); (iv) profit motives vs. altruism of human behaviour; (v) loss of animal genetic diversity and integrity of species; and (vi) altering of natural course of animal evolution. Raising calves in the dark and feeding them with diet, for developing high quality veal, and battery rearing of broilers have been considered as unethical. 2.Public policy Animal biotechnology is expected to have significant impacts on food production and processing. While supporters predict several economic, social, and environmental benefits, critics raise concerns about the bio-safety and ethics of biotechnology, including concern about whether governments (National and World Trade Organization) can adequately regulate the applications of biotechnology (Kendall and Hoban 1994). In fact, animal biotechnology has become an important, but controversial public policy issue in several countries. Since consumers will make the ultimate decisions about the acceptability of animal products generated through biotechnology, efforts should be made to educate the public about several applications and issues associated with animal biotechnology so that consumers can express informed consent. Instead of providing justification for such incidences, we should have an ethical code of conduct using animals for bio-medical research and development. 3.Regulatory mechanisms The regulatory mechanisms related to the application of animal biotechnology are highly influenced by our perceptions of the biotechnological products. Some of us believe that such products are articles of food, food additives, animal drugs, animal biologic, laboratory animals, or are manufacturing sites for human drugs, biologics, and devices. Other people may feel that animal biotechnology is a simple extension of selective animal breeding, a technology for introducing pharmacologically active substances into animals, a source of improved farm income, a potential environmental problem, an ethical conundrum, cruelty to animals and a case for animal rights, even to the extent of demanding/putting/an embargo on the use of animals for research purposes (Matheson, 1994). Recent case of freeing the animals (monkeys at NII) by NGO, Blue cross, is another extreme case of animal rights activism. Instead of providing justifications for such incidences, we should have an ethical Code of Conduct on the use of 17

animals for bio-medical research and development or based on these positive and negative considerations (Matheson, 1994). In any event any policy decision should not lead to slowing down of R&D efforts aimed at human and animal welfare. Consequently, formulating of the national/international animal biotechnology policy cannot be taken as an extension of such issues related to plant biotechnology; of course some issues may require common answers. General public, socio-economists, animal rights activists, religious groups, scientists and government officials (national, international organizations, WTO) differ in their perceptions of the use of products generated through animal biotechnology. Animals are capable of producing drugs, milk with medical food chains, human biologics and food products. What is our attitude towards such innovations and their products? 4.International developments While several countries, which use animal biotechnological products and/or have international trade in such products, have formulated regulatory mechanisms, we in India, have done so little in this direction. Cross country surveys indicate that there are several approaches to the regulatory mechanisms, each with advantages and disadvantages, but we, in India, should formulate regulatory rules and legislation appropriate to our socio-economic and religious attitudes. A consistent approach for similar products on an international basis is certainly desirable and may ultimately emerge under the banner of WTO rules and regulations. It is also desirable to work on an International Code of Conduct on the applications of animal biotechnology as they affect animal genetic resources. This should be in line with a similar International Code on applications of plant biotechnology on plant genetic resources, under the banner of FAO Commission on Genetic Resources for Food and Agriculture. The author has continued to assist the Commission in the preparation of this code since 1990 through two consultancies and correspondences. The Indian National Academy of Agricultural Sciences through this workshop may make not only a recommendation but also prepare a draft Code of Conduct on the use of animal biotechnology. This code may eventually become a negotiating draft for an International Code of Conduct on Animal Biotechnology. The author can assist the National Academy in this noble endeavor. Advantage can also be taken from the guidelines and issues covered in the FAO Code of Conduct for Responsible Fisheries (Anon, 1995). The Codex Alimentarius Commission (CAC) of the FAO & WHO has prescribed over 200 standards, 45 18

codes of practices and 2000 maximum limits for residues of agricultural and veterinary chemicals (Anon, 1999). The CAC is presently developing Recommendations for the Labeling of foods obtained through biotechnology. The CAC is also considering the development of general standards, which would apply basic food safety and food control disciplines to food derived from biotechnology. Elements of concern are potential allergenicity, possible gene transfer from transgenics, pathogenicity deriving from the organisms used, nutritional aspects and labeling. The process of risk analysis should also be applied to biotechnology and food safety. Under the WTO also, its SPS Agreement (Agreement on the Application of Sanitary and Phytosanitary Measures) covers measure in trade that are intended to protect human, animals and plant health or life. These international developments should be taken notice of in formulating our national policy on biosafety of animal biotechnological products. 5. ETHICS AND ANIMAL WELFARE IN ANIMAL BIOTECHNOLOGY FOR FARM PRODUCTION

Ethics: intrinsic arguments against animal biotechnology


Ethical concerns about the genetic engineering of animals may conveniently be divided into two basic types. The genetic engineering of a living organism may for a variety of reasons be thought of as being morally problematic in itself; for example, such engineering may be perceived as wrong or morally dubious due to the mode of production or to the source of genetic material. In such cases the author will use the term intrinsic concerns. But genetic engineering may also be thought of as morally problematic because of its consequences. In that case, the term extrinsic concerns will be used in this paper. When discussing the ethical issues of GM animals it is important to keep these two categories apart. For instance, considerations of risk are often central to extrinsic concerns, but have no force whatsoever where objections to a procedure are based on fundamentally intrinsic concerns. Comstock (9) argues that all the variants of intrinsic arguments against (animal) biotechnology could be summarised in the following claim: it is unnatural to genetically engineer plants, animals and foods. Such an argument could be spelled out in more detail in various ways. This paper will consider three of the most salient ones.

We should not be playing God


Some people hold that it is ethically questionable to transfer genes from one species to another. This attitude is sometimes grounded in a religious belief that humankind should not violate boundaries set by God, and so this line of reasoning is usually referred to as the playing-God argument (8, 20). Any re-designing of nature through the insertion of new genes is morally unacceptable according to this view. The argument does not occur in the Bible (in fact one may cite statements that contradict it), but is based on an interpretation of Gods will. If one accepted the playing-God argument, this would entail an unconditional rejection of the genetic modification of fish and other farm animals. The basic assumptions of the argument are the following: God has drawn up invisible boundaries between the realm of God and the realm of humans. Those that transcend this boundary are guilty of hubris (excessive pride). Obviously any such argument depends on specific religious assumptions about the relationships of God, humans and animals, and the argument is generally proposed by adherents of some forms of Christianity. The problem here is to know where the ethical boundary lies. The only safe assumption would be not to interfere with the course of nature at all, but this would arguably exclude all systematic breeding efforts through the whole sphere of agriculture,

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particularly all selective breeding of animals. If, however, society wants to allow for breeding, then people must presumable regard some changes in the genetic make-up of species as ethically acceptable. In this regard the results of selective breeding and the results of recombinant DNA techniques are on a par.

Genetic modifications of animals break down natural species boundaries


Intrinsic ethical concerns of a similar kind may also be raised without relying on an explicit religious foundation. Respondents in surveys about biotechnology (e.g. in the Euro barometer surveys conducted on behalf of the European Commission) (2, 17) often express concern that genetic engineering is unnatural. The argument then is that the genetic modification of animals breaks some natural order among species that is in a finely tuned state of equilibrium. Nature and all that is natural is then assumed to be valuable and good per se; tampering with nature, as do all forms of genetic engineering where species boundaries are crossed, appears as a hubristic interference with the natural order, and is thus conceived as intrinsically wrong. Any such argument faces two difficulties that need to be examined. First, what is meant by being natural or unnatural, and second what is good about being natural and, conversely, bad about being unnatural? It seems quite clear that a large part of our food production represents some form of unnatural element in nature. All domestic species and most agricultural crops are the results of selective breeding. Most of the nature surrounding our cities has been shaped by human intervention and is, therefore, far from being natural. Thus a notion of nature in a general sense seems at least problematic when used in this argument. More specifically, it may be argued that it is the genetic boundaries that exist between species that constitute what is naturally given and that need to be respected. The insertion of unrelated genes may change the entire direction of evolution and represent human goals rather than a natural development. Against this view is the argument that species are never static: their genetic makeup changes over time. It can also be claimed that species are not genetically isolated. Certain viruses, for example, carry genetic material from one species to another. Also the notion of species may not be so firmly established from a biological point of view as many seem to assume; there are alternative conceptions of what a species is. In sum, the reference to nature and the natural is not such a firm foundation as the argument seems to presuppose. It merely points to the essential social construction of the term nature.

Animal biotechnology implies the commodification of all life forms


The main intuitive argument here would rest on the reductionist nature of genetic engineering. This reductionism implies that living organisms are treated the same way as all other parts of nature that become the object of technoscientific intervention: in other words, life-forms are seen as nothing other than complicated machines (9). The complaint is then that such an approach implies a disrespect for life. Life is of a higher value than can be measured in economic terms alone. The problem with this kind of reasoning is parallel to the arguments mentioned before. Generally speaking, if this argument were true and taken as a general and unconditional principle for the moral acceptability of interventions in nature, then a lot of other activities would fall prey to it activities that are normally regarded as morally acceptable. Food production with animals that are not GM implies a commodification of life and in some sense reduces the organism to an economic value, as does all trading in living organisms. The fact that the assumed moral principle leads to what many would regard as counterintuitive consequences namely, that these practices are morally wrong would then be taken as evidence that the principle fails, at least if taken to its logical conclusion. Therefore, even if the assumption that genetic engineering entails a commodification of life forms is true, this in itself would not be sufficient to reject animal biotechnology in general. The other ethical aspect to be considered in this paper covers the extrinsic arguments against animal biotechnology: arguments about the consequences rather than the nature of our technology. One such line of argument relates to animal welfare, another to environmental considerations.

Extrinsic arguments against animal biotechnology


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Animal welfare
There are different schools in regard to animal welfare, some more restrictive than others. Some take as their only reference point the feelings of the animal, so that the ability to feel pain becomes the crucial factor (11). Others look at animal health and physiological functions (7) as crucial criteria for defining animal welfare. Others again (such as those propagating eco-farming) regard the conditions of the animals natural living environment as the foundation of animal welfare, so that the ability to follow natural instincts in a biologically preferred environment becomes crucial. These differences among competing conceptions are only partly based on science. To a considerable extent the differences are due to different philosophical outlooks, and may not be reconcilable. This is not altogether surprising, since views of animal welfare obviously depend on a variety of implicit value assumptions. In Europe, the 1965 United Kingdom report on animal welfare that became known as the Brambell report was highly influential. The Brambell report (6) included the well known five freedoms: a) freedom from hunger and thirst by ready access to fresh water and a diet to maintain full health and vigour b) freedom from discomfort by providing an appropriate environment including shelter and a comfortable resting area c) freedom from pain, injury or disease by prevention or rapid diagnosis and treatment d) freedom to express normal behaviour by providing sufficient space, proper facilities and company of the animals own kind e) freedom from fear and distress by ensuring conditions and treatment that avoid mental suffering. While it would certainly be wrong to claim that these freedoms have actually been translated into national law, one might still hold that they set a soft-law standard that is often referred to and that some countries have aimed to include in their national regulations for farm animals. Like all such general principles, these freedoms are imprecise and leave room for different interpretations. Much effort has therefore been devoted to specifying what phrases such as sufficient space might mean for specific species in animal production, e.g. battery hens. It is not too farfetched to say that these animal freedoms function in much the same way as the Charter of Human Rights does for humans. They define a prima facie ethical standard that informs our judgements of particular cases. Indeed, there are striking parallels between the five freedoms of the Brambell report and the celebrated hierarchy of needs of the psychologist Abraham Maslow (24), who defined a five-level scale of basic human needs. The five freedoms seem loosely to match this thinking. In regard to animal biotechnology, the challenge is certainly not less than the challenge of assessing animal welfare in general. The crucial question seems to be whether the genetic manipulation of the animal affects properties relevant to animal welfare. Many would consider that if there were negative effects on animal welfare the probability of a positive regulatory decision on the production and marketing of GM animals would be severely reduced.

Animal welfare in regard to transgenic animals


It is difficult to say anything specific about the welfare of transgenic animals in general. In principle it may be better or worse than the welfare of the original animals. Obviously, a lot will depend on the kind of modification that has been made: in other words, which genes have been modified. Thus research will be needed to document the effects of such modifications. Even though the principal responsibility for documenting welfare effects may rest with the company that seeks to market the product, the availability of independent research scrutinising the effects is likely to be crucial. This will be a consequence of the lack of trust that the public now expresses towards industrial research. However a range of experience is already available in regard to transgenic fish. It has been claimed that unexpected phenotypical disadvantageous changes are the rule rather than the exception in the genetic modification of fish (33). More specifically, deformities of the head and other parts of the body have been documented for transgenic salmon (Oncorhynchus kisutch) (10). The documented deformities obviously have serious welfare implications for the fish. In addition, morphological changes and changed allometry have been documented for the same fish, leading to reduced swimming abilities (13, 23, 29). It also seems that transgenic salmon show deviant behaviour in the sense that there is an increased level of activity in regard to feed-intake and swimming (1, 10). Thus one can claim that the literature documents changes in morphology, physiology and behaviour in transgenic salmon.

Environmental effects of animal biotechnology 21

Some animal biotechnology may have negative effects on the environment and biodiversity. As a general rule, it is probable that the larger the animal and the better the containment, the lower the risk of unintended environmental effects. This relates to the ease with which processes can be controlled. Since the ecological interactions of species may be quite complex and not always easily or quickly detectable, such a rule cannot be applied in all cases. Leaving aside other environmental effects of feeding and containing the animals in appropriate surroundings (effects that may not be specific to animal genetic biotechnology), the main concern today is the spread of genetic material to wild relatives. Disturbances in our ecosystem raise ethical concerns, especially when some of the affected species and populations may already be at risk of extinction. Again, the main studies of these effects have been undertaken in regard to transgenic fish. One way to approach the possible gene flow from one population to another is to assess the net fitness of a specific type of fish (30). Muir and Howard (26, 27) list six traits that determine the net fitness of any animal, including transgenic fish: juvenile viability: chances of surviving to sexual maturity adult viability: chances of surviving to procreate fecundity: number of eggs produced by a female fertility: percentage of eggs successfully fertilised by male sperm mating success: success at securing mates age at sexual maturity. Rather than regarding these as independent or associated factors, scientists now develop mathematical models to combine these parameters of the organisms net fitness in order to assess the genetic consequences of escapes of a transgenic organism into the environment. Three possible scenarios or hypotheses can be discerned (30).

The purge scenario


When the net fitness of a transgenic organism is lower than that of its wild relatives, natural selection will work to purge any transgenes that the wild relatives may inherit (21). Some transgenic fish may indeed fall under this scenario, depending on the kind of modification that has been done. It is claimed, though, that this scenario need not be without any long-term effects. In small populations even temporary declines in net fitness are said to potentially threaten the survival of the population.

The spread scenario


A 2003 report by the Pew Initiative on Food and Biotechnology stated that when the net fitness of a transgenic fish is equal to or higher than the net fitness of a wild mate, gene flow is likely to occur and the genes of the transgenic fish will spread through the wild population (30). Some transgenic fish, such as growth-enhanced Coho salmon, reach sexual maturity earlier than their wild counterparts, and this has a significant effect on net fitness. In this scenario the invasion of transgenic fish has a lasting genetic effect on the wild counterparts.

The Trojan gene scenario


In this model it is suggested that enhanced mating success coupled to reduced adult viability would result in a rapid decline of the wild population. The transgenes would quickly introduce their genetic material into the wild population, while the lower viability would reduce population sizes. Thus, interbreeding of transgenic organisms with wild relatives would cause a rapid decline in total population. Again there are a number of significant uncertainties relating to the effects on the environment and biodiversity. Obviously, no general or definite answer is possible at this stage of our knowledge. One may point out that factors such as the introduction of sterile transgenic species could change the potential environmental threats significantly. Even in these cases, however, instability and uncertainty are factors to be taken seriously in our assessments. While transgenic fish may pose an environmental threat of serious proportions due to the lack of control of escapes, the principal points apply to all animal biotechnology. The experiences of exotic species can serve as a rough model for some of the potential threats.

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6. CONCLUSION The next decade will be an extremely exciting time for agriculture and the feed industry. The production of genetically enhanced plants, feed additives, and foods will affect our lives in many ways. Biotechnology will greatly affect the field of medicine, environmental protection, food production, and agriculture. The advantages of using modern biotechnology for the production of animal feeds will be apparent for everyone from the farmer to the consumer. Two key factors will drive public opinion regarding current and future use of GMOs in our food chain. First, there is no way to produce the quantities of food needed for the rapidly increasing world population using standard backcrossing genetic techniques with tillable land available. Therefore, it is must to use the new genetic tools available to produce the GMOs needed to feed our fellow man. Second, I think everyone in the feed and animal industry as well as the general public is concerned about improving our present and future ecological situation. The first wave of feedstuffs already commercially available, that are considered transgenic, will allow farmers to use less chemicals to control insects and use various no-till soil conservation farming methods with herbicide resistant cultivars.

Modern biotechnology also used to decrease problems of dry matter waste, odor, and too much phosphorus and nitrogen in animal waste. Hence, biotechnological interventions can play a vital role not only in the productions but also in environmental protection and sustainability. Therefore, an integrated approach to livestock development would be required, as no single option would provide the answer for achieving the goal in an eco-friendly, cost effective and sustainable manner.

Recommendations
Companies are distrusted as information source all over Europe (Eurobarometer, 1997). This distrust against companies cannot be overcome with more information from the companies. Instead, companies can only improve the way they are viewed upon, through working on their own values and norms. With the primary goal of clarifying them for themselves, i.e. for the people acting in the name of the company. The idea is that clearly defined corporate values and ethical principles would in fact serve as a common guide to decision-making within the company. This, in turn, would enable external stakeholders (the consumers, but also other institutional actors) to better evaluate the companys choices enhancing the understanding of 23

both the scientific, the economic and the moral rationale behind them. It is therefore recommend that biotechnology companies start to make their own moral codes more explicit, undertaking such a process in a transparent way, and being open to dialogue with concerned stakeholders, such as consumers, citizens and environmental groups, the scientific community, regulatory bodies and the media. In order to overcome the strict virtue-ethical or deontological judgement of the public, companies could give other actors a role in the communication with the public. This can be done by intermediate organisations. Another possibility is to invite consumer, environmental or other issue-organisations in the company and to start low profile interaction (as practised by Novo Nordisk in Denmark). However, they should keep in mind the virtue-ethical judgement mentioned above. Starting communication is something they cannot do on their own: it still takes two to tango.
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The fact that companies do better communicate with governmental bodies for instance in the process of developing regulations, than with the public, suggests that companies make the same mistake as the regulators. This is to say that a mismatch between concerns on risk and safety of companies and the public moral questions may be the cause for the lack of communication between public and companies. Therefore it is recommend that - given the need and explicit wishes of companies to communicate -, further research on the causes for this difference in communication is needed. This research should focus on different conceptions of risk and different conceptions of morals in the companies, in issue-groups and in regulatory bodies. The attitude towards biotechnology differs strongly over countries. Also the differences in national debate on biotechnology are surmount. Companies behaviour strongly reflects their social and cultural environment (at least on the point of morals). This means that the differences in moral competitiveness between companies are also (caused by) differences between national cultures and that moral competitiveness of companies is strongly influenced by the national culture. Companies should keep these differences in mind when they try to start communication processes in other countries. This also means that when biotech companies want to implement the EuropaBio principles of dialogue and communication they should adopt it to the characteristics of the national debate on biotechnology.

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