In This Issue:
What's in the Pipeline?
A Reader Responds
A Word from USDA's Biotechnology Information Center
Virus Resistant Papayas Deregulated
A Review of Transgenic Aquatic Organisms
Antifreeze Transgenes Increase Cold Tolerance and Freeze Protection
Will EASDAQ Help Growth of European Agbiotech Firms?
As this summer's field tests are winding down to harvest and termination, planning is well underway for the next round. As of September 13, dozens of notification letters and permit requests for off-season trials have been received at USDA/APHIS's Biotechnology Permits unit, and no doubt more are on the way.
A quick survey of submissions reveals a lot of overlap in the development of herbicide tolerant and insect resistant corn lines. Male sterility, which may be suitable for biological confinement to prevent gene flow through pollen transmission, will be evaluated in rapeseed, Brassica oleracea, and corn. Antibiotic-producing soybeans and rapeseed that produces a pharmaceutical protein are also scheduled for field testing.
Under the notification procedure, crops having the following traits will be tested in warm, sunny fields in Puerto Rico and/or Hawaii, except where noted (acknowledgment of some notifications may still be awaiting state response):
Cotton:
Permits are pending for field tests of:
Connect to the USDA/APHIS/BBEP Biotechnology Permits homepage www.aphis.usda.gov/bbep/bp/ for updated information on field tests and product deregulations.
Pat Traynor, Editor
A READER RESPONDS
I was most interested and pleased to see Dr. Stewart's comment, as I believe a national discussion of how our regulatory agencies should do risk assessment of biotechnology is long overdue. I'm a toxicologist working for a regional office of the Environmental Protection Agency. My specialty is human health risk assessment, and I've become very interested in the methodologies being used to assess the risks of this novel technology.
Several observations:
1. Dr. Stewart rightly suggested that regulatory agencies should
critically analyze risk assessment data and take to heart
recommendations from independent scientists. But there is a much more
fundamental need. For biotechnology there are no commonly accepted risk
assessment protocols, guidance, methodologies, testing guidelines or
science policies, as we have with human health risk assessment.
There is in place today a systematic approach to human health risk assessment because in 1983 the National Academy of Science conducted a study of the institutional means for risk assessment (published as "Risk Assessment in the Federal Government; Managing the Process"). It became the basis of a common framework for human health risk assessment now used by all federal agencies.
A similar framework is now needed - indeed, is overdue - for assessment of products of biotechnology. Assessments are done on an ad hoc basis by different agencies (EPA, FDA, USDA) and within at least EPA, by different programs. Obviously, this creates assessments which are not comparable, and are of variable quality. Assessors working independently and using different protocols are not sharing insights, raising common issues or learning from one another how to address the unique challenges raised by this new technology.
General principles, such as: "Avoid the commercialization and large-scale release of any crop engineered with fitness-altering genes and capable of interbreeding with wild relatives" (Dr. Stewart's excellent wording) are needed not only to protect the environment, but to guide manufacturers in their applications, and to protect the industry from the stigma of ill-conceived products.
2. Dr. Stewart touched on the costs and benefits of taking the avoidance vs management/ mitigation paths. The question of who bears the costs and who reaps the benefits is interesting and not always simple. True, the manufacturer must pay substantial research and development costs, but the likely far larger costs of any environmental or human health disasters will be borne by society. The manufacturer may reap a substantial economic benefit if its product succeeds, but of course this is an economic risk. On the other hand, economic success for an individual company does not necessarily assure societal benefits. Under the Toxic Substances Control Act (TSCA) for example, efficacy of a product is not required for approval of an application; in fact if a company could successfully market an ineffective product, their projected profits would be counted as the benefit to be weighed against potential environmental risks. EPA is facing this situation right now as it ponders approval of a genetically engineered nitrogen-fixing soil bacterium with questionable efficacy.
3. As Dr. Stewart pointed out, there is tremendous economic pressure to develop new products for short-term gain, without thought about the long-term consequences. There is also tremendous political pressure: to help small businesses, to make the U.S. a technological leader, to keep government out of the private sector, to reduce regulations. The management of our regulatory agencies feels that pressure. I hope that scientists such as Dr. Stewart will take advantage of the respect accorded them, and offer their views to a larger audience, so that political pressure can be balanced with reasoned concerns.
Suzanne Wuerthele
EPA Regional Office, Denver, CO
wuerthele.suzanne@epamail.epa.gov
How do I learn about research related to genetically engineered algae? What biotech-related databases are available online? What organizations are involved in biotechnology education? These are just a few of the thousands of questions handled each year by the Biotechnology Information Center (BIC) in Beltsville, MD. The BIC is one of 10 information centers located at the National Agricultural Library, which is part of USDA's Agricultural Research Service. Now in its 11th year of operation, the BIC is regarded as one of the foremost sources of information about agricultural biotechnology around the world.
The BIC staff is familiar with concepts and techniques used in biotechnology and will perform complimentary searches of the AGRICOLA database on specific topics or conduct an exhaustive search of major databases on a cost recovery basis. In addition to developing aids to assist consumers, educators, researchers, and policy makers, the Center's Internet website at www.nal.usda.gov/bic provides access to an extensive collection of biotechnology documents and links to other sites. In any given month, users from more than 30 countries connect to the site, accessing up to 19,000 documents. One of the most popular areas of the BIC site provides access to full-text biotechnology patents.
Much of BIC's strength lays in its commitment to continually updating all of its resources, guides, bibliographies, and databases. The "Quick Bibliographies", commonly called QB's, provide customers with a list of all the latest articles, papers, books, and monographs pertaining to a particular subject area, thus saving the patron hours of time spent in library research. Most recently, BIC added the following QB's to its list: Risk Assessment/Biosafety; Public Perception; Gene Gun/Biolistic Technology; Bioethics, Legislation and Regulation; Patenting Issues; Commercialization and Economic Aspects; Herbicide Tolerance/Resistance in Plants; Viral Resistance in Plants/Viral Coat Proteins; and Education and Training. These titles are now or will be soon available via BIC's webpage.
Another BIC specialty is its extensive collection of biotechnology-related audio-visuals and audio-tapes. All are available for loan to any individual within the United States through a standard interlibrary loan request. Some of the latest additions to this collection include, "Licensing Veterinary Biologics: Traditional and Recombinant Vaccines", "Whither Biogenetics?", "Designer Plants", and "Superanimals, Superhumans?".
To learn more about the Biotechnology Information Center resources and services, call 301-504-5947 or email: biotech@nalusda.gov.
The first genetically engineered perennial crop was cleared for commercial production September 5, when USDA/APHIS biotechnology staff concluded that two lines of papaya trees modified with the coat protein gene from papaya ringspot virus (PRV) are just as safe to grow as papaya cultivars developed through traditional breeding practices. The trees were developed by researchers at Cornell University and the University of Hawaii.
Papaya ringspot virus is the most important pathogen of payaya on a worldwide basis; all major production areas are affected and the virus continues to spread into new areas where papayas are grown. PRV, a member of the potyvirus group transmitted by aphids, is a tenacious pathogen that has never been successfully eradicated once introduced. A 1992 outbreak of PRV in Hawaii's Puna district, where 95% of the state's papayas are grown, has become so widespread it is unlikely that it can be contained by control measures.
Genetic resistance has been identified in papaya germplasm but the trait is polygenic, only moderately effective, and not found in cultivars suited to Hawaii. Cross-protection by deliberate inoculation with a mild mutant strain of PRV (HA 5-1) to prevent subsequent infection by more virulent strains offers some protection against Hawaiian strains of PRV. However, while cross-protected trees have only relatively mild disease symptoms, yield is reduced 10-20% and the approach requires inoculation of every seedling generation.
These limitations were circumvented by transforming the cultivar 'Sunset' with the coat protein gene of mild PRV strain HA 5-1. After initial tests in the greenhouse, a two-year field test of transformed line 55-1 showed that R0 plants and their progenies are highly resistant to Hawaiian isolates of PRV; the line is susceptible to virus isolates from other parts of the world, including Thailand. Greenhouse tests of transformed line 63-1 indicate resistance to Hawaiian PRV isolates and better resistance to Thai isolates than line 55-1.
The APHIS determination that transgenic 'Sunset' papaya lines 55-1 and
63-1 shall no longer be regulated is based on their conclusions that
the transgenic lines:
According to Dennis Gonsalves, principal investigator from Cornell University, growers in Hawaii are anxious to get the new papaya lines planted.
Pat Traynor
Information Systems for Biotechnology
traynor@nbiap.biochem.vt.edu
Gene transfer techniques have been applied to a large number of aquatic organisms, both vertebrates and invertebrates. Gene transfer experiments have targeted a wide variety of applications, including the study of gene structure and function, aquaculture production, and use in fisheries management programs. This article briefly reviews the status of development of transgenic aquatic organisms.
Because fish have high fecundity, large eggs, and do not require reimplantation of embryos, transgenic fish prove attractive model systems in which to study gene expression. Transgenic zebrafish have found utility in studies of embryogenesis, with expression of transgenes marking cell lineages or providing the basis for study of promoter or structural gene function. Although not as widely used as zebrafish, transgenic medaka and goldfish have been used for studies of promoter function. This body of research indicates that transgenic fish provide useful models of gene expression, reliably modeling that in "higher" vertebrates.
Perhaps the largest number of gene transfer experiments address the goal of genetic improvement for aquaculture production purposes. Most experiments entail transfer of genes for growth hormone (GH) or other growth factors. Devlin and coworkers reported the most dramatic results to date, with transgenic coho salmon families exhibiting growth rates 11 times those of non-transgenic controls (1). However most studies, such as those involving transgenic Atlantic salmon and channel catfish, report growth rate enhancement on the order of 30-60%. In addition to the species mentioned, GH genes also have been transferred into striped bass, tilapia, rainbow trout, gilthead sea bream, common carp, bluntnose bream, loach, and other fishes.
Shellfish also are subject to gene transfer toward the goal of intensifying aquaculture production. Growth of abalone expressing an introduced GH gene is being evaluated; accelerated growth would prove a boon for culture of the slow-growing mollusk. A marker gene was introduced successfully into giant prawn, demonstrating feasibility of gene transfer in crustaceans, and opening the possibility of work involving genes affecting economically important traits. In the ornamental fish sector of aquaculture, ongoing work addresses the development of fish with unique coloring or patterning. A number of companies have been founded to pursue commercialization of transgenics for aquaculture. As most aquaculture species mature at 2-3 years of age, most transgenic lines are still in development and have yet to be tested for performance under culture conditions.
A number of experiments utilize gene transfer to develop genetic lines of potential utility in fisheries management. Transfer of GH genes into northern pike, walleye, and largemouth bass are aimed at improving the growth rate of sport fishes. Gene transfer has been posed as an option for reducing losses of rainbow trout to whirling disease, although suitable candidate genes have yet to be identified. Richard Winn of the University of Georgia is developing transgenic killifish and medaka as biomonitors for environmental mutagens, which carry the bacteriophage phi X 174 as a target for mutation detection. Development of transgenic lines for fisheries management applications generally is at an early stage, often at the founder or F1 generation.
Broad application of transgenic aquatic organisms in aquaculture and fisheries management will depend on showing that particular GMOs can be used in the environment both effectively and safely. Although our base of knowledge for assessing ecological and genetic safety of aquatic GMOs currently is limited, some early studies supported by the USDA biotechnology risk assessment program have yielded results. Data from outdoor pond-based studies on transgenic catfish reported by Rex Dunham of Auburn University show that transgenic and non-transgenic individuals interbreed freely, that survival and growth of transgenics in unfed ponds was equal to or less than that of non-transgenics, and that predator avoidance is not affected by expression of the transgene.
Laboratory studies of transgenic medaka by Bill Muir and colleagues at Purdue University indicated that large males gain a higher frequency of matings, but that transgenic offspring exhibit decreased viability; computer modeling suggests that the demographic viability of medaka populations could be threatened by introduction of transgenics. Possible impacts of monosex or triploid grass carp stocks will be assessed by Bill Shelton of the University of Oklahoma; although these carp will not be transgenic, the results will have bearing on use of transgenic stocks as insights will be gained on the efficacy and environmental safety of these means of reproductive confinement.
Given the desire to pursue research and development with genetically modified aquatic organisms in the face of incomplete knowledge regarding environmental risks, a decision support tool for assessing and managing risks was developed. The Performance Standards for Safely Conducting Research with Genetically Modified Fish and Shellfish was produced by a working group under USDA's Agricultural Biotechnology Research Advisory Committee with input from a wide range of aquatics professionals (2). The Performance Standards serve as a guide for a researcher to assess potential risks associated with a proposed experiment with an aquatic GMO, and to adopt appropriate confinement should any risk be identified. Documented completion of the performance standards should help the researcher gain approval for the experiment from an institutional biosafety committee. The Performance Standards have been converted into a computer software package, which can be obtained without cost by downloading it from the NBIAP website at http://www.isb.vt.edu, by e-mail request at nbiap@vt.edu, or by calling 540/231-3747. Print copies are available from the Biotechnology Information Center of the National Agricultural Library (see article elsewhere in this issue).
References
1. Devlin, R.H., T.Y. Yesaki, E.M. Donaldson, and C.-L. Hew. 1995.
Aquaculture 137:161-169.
2. Agricultural Biotechnology Research Advisory Committee. 1995.
Performance Standards for safely conducting research with genetically
modified fish and shellfish. Part I. Supporting text. Document no.
95-04. Part II. Flowcharts and accompanying worksheets. Document no.
95-05. National Agricultural Library, Beltsville, MD.
Eric Hallerman
Virginia Tech
ehallerman@vt.edu
Marine fish that live in icy waters, such as wolf fish and sea raven, often synthesize antifreeze proteins (AFPs) to protect against freezing. Transfer of AFP genes into economically important fish species has been proposed as a method for conferring freeze protection. Production of AFP-expressing strains of salmon and other fish, for example, would promote the development of sea-pen aquaculture in colder regions of the world.
Experiments to introduce AFP genes suggest that genetically stable germ-line transformed fish can be produced (1). Integration of a winter flounder AFP gene into the genome of the Atlantic salmon following microinjection of eggs was accompanied by low level expression in a small number of fry. Inheritance of the AFP gene was stable, and about 50% of the offspring resulting from transgenic F1:wildtype crosses contained the gene. When goldfish oocytes were microinjected with ocean trout AFP genes, nearly a quarter of the resulting 2-month old fish contained one to several copies of the transgene. More than 50% of the offspring of separate crosses between two transgenic males and a control female inherited the gene, and AFP was detected in both F1 and F2 progeny. Transgenic goldfish experimentally challenged with low temperatures also had significantly greater tolerance for cold conditions than controls, implying that the AFP gene product may provide cold tolerance as well as freeze protection.
Transgenic expression of fish AFP genes may also be a means of increasing the frost resistance and freeze tolerance of plants (2). Tobacco plants transformed with the winter flounder AFP gene produced mRNA, demonstrating that the gene was transcribed, but no detectable AFP protein was produced. However, protein was detected after cold exposure (4 C for 24 h), suggesting that this may be a suitable strategy for producing frost resistant plants.
References
1. Molecular Marine Biology and Biotechnology, vol. 1, 1992,
p.309-317; vol. 4, 1995, p. 20-26.
2. Plant Molecular Biology, 1993, p.377-385.
J. Glenn Songer
University of Arizona
gsonger@ccit.arizona.edu
WILL EASDAQ HELP GROWTH OF EUROPEAN FIRMS?
It is well known that one of the more onerous tasks facing the biotechnology industry is the relentless pursuit of capital to fund research and development. This has been particularly difficult for European firms due to the lack of a European stock exchange which catered to high-risk technology firms in industries like biotech. This month a new option has arisen in the form of the European Association of Securities Dealers Automated Quotation (EASDAQ) stock exchange.
The pan-European EASDAQ is an initiative of the European Association of Securities Dealers (EASD) and the European Venture Capital Association. One of the major incentives behind the development of the EASDAQ was the need to fill a void that has left most of Europe's smaller growth companies without access to liquid public markets. It is estimated that only 2.2% of European companies with less than 500 employees have raised capital on a stock exchange. This lack of public equity markets has also had an impact on the venture capital community's ability to raise funds. All of this is in contrast to the situation in the United States, where the National Association of Securities Dealers Automated Quotation market (NASDAQ) boasts listings of over 4,900 companies. The NASDAQ has been a significant source of capital for biotechnology firms in the U.S. and abroad, and the hope of many is that the EASDAQ will ultimately play the same role for European biotechnology. One study conducted by Coopers & Lybrand indicated that as many as 500 stocks (including biotechnology stocks) could be traded on the EASDAQ within 5 years, representing firms from most of the major European countries (1).
Not everyone is so enthusiastic about the prospects for the new European exchange. A recent article in Nature Biotechnology questions whether or not the need for such an exchange still exists in Europe, as was the case when the initiative was first put forth several years ago. For one thing, since the time that the EASDAQ concept was proposed in 1994, the London Stock Exchange (LSE) has revised its listing rules, making it easier for high-tech companies such as biotechnology firms to enter the market. In addition, other small-cap markets have reemerged including the Alternative Investment Market (AIM) in London and the Nouveau March‚ in Paris. Finally, a number of European firms have accessed capital through the NASDAQ, which the EASDAQ ultimately hopes to leverage through a relationship with NASDAQ offering a dual listing to firms listed on either exchange.
Those concerned about the value of the EASDAQ fear that there simply may not be enough buying and selling of stock to provide adequate commissions to brokers who provide access for large institutional investors. One possible outcome that has been mentioned is that the EASDAQ will be used more by non-UK firms who have less access to domestic markets, while UK-based firms will tend to list on the LSE or AIM. Time will tell how many other biotechnology firms will follow in the footsteps of the European biomedical diagnostics firm Innogenetics, the first European bioscience firm to be quoted on the new exchange (2).
References:
1. EASD home page on the Internet World Wide Web at http://ourworld.compuserve.com/homepages/easd/
2. Ward, M. EASDAQ opens, with some unease. Nature Biotechnology,
Vol. 14, No. 9, September 1996, pp. 1075-1076.
William O. Bullock
Institute for Biotechnology Information, LLC
Research Triangle Park, NC
http://www.biotechinfo.com
The material in this News Report is compiled by NBIAP's Information Systems for Biotechnology, a joint project of USDA/CSREES and the Virginia Polytechnic Institute and State University. It does not necessarily reflect the views of the U.S. Department of Agriculture or of Virginia Tech. The News Report may be freely photocopied or otherwise distributed without charge. P.L. Traynor, Editor.
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