Analysis of Results: This is the focus of much of ARS' research related to food safety and the security of the U.S. agricultural production system. Under Goal II, 59 Indicators are aligned under 10 Performance Goals. Because of the unique and dynamic nature of research, several Indicators were added, deleted, or modified in this Report that did not first appear in the Annual Performance Plan for FY 2001. This was done to ensure that significant accomplishments that were not anticipated last year were reported. While it is not possible to report research accomplishments numerically, the progress projected in all 59 Indicators was completed or substantially completed during FY 2001.

OBJECTIVE 2.1: Secure food and fiber system: "Maintain a safe and secure food and fiber system that meets the Nation's needs now and in the future."

STRATEGY 2.1.1: Plant and animal production systems: Improve efficiency of agricultural production systems to ensure the security of the Nation's food, fiber, and energy supply.

During FY 2001, ARS will

ACCOMPLISHMENTS: Appropriate nesting material is the single most important factor preventing the widespread use of the blue orchard bee as a pollinator of almonds, cherries, pears, and other orchard crops. Researchers at Logan, Utah, in collaboration with a cherry producer in North Ogden, Utah, have developed an inexpensive, durable, and easily-sanitized nesting block for this purpose.

IMPACT/OUTCOME: The new nesting block will likely be adopted by orchard growers.

ACCOMPLISHMENTS: New vaccines have been developed by several scientists at various laboratories against foot and mouth disease, respiratory disease in cattle, and swine influenza.

IMPACT/OUTCOME: These new vaccines will help producers combat losses from diseases where it exists and increase preparedness for foreign diseases should incursion occur (e.g., foot and mouth diseases).

ACCOMPLISHMENTS: Results of research concerning seed production, establishment, fertilizer strategies and grazing management of old world bluestem grasses at Woodward, Oklahoma, make it possible to triple livestock production per acre. Research also has demonstrated that rangeland net returns can increase between $6 to $12 per acre by significantly integrating these lands with old world bluestem pasture and croplands to create a year-around grazing system.

IMPACT/OUTCOME: Profitable agricultural production depends on lowering input costs while maintaining productivity and protecting the environment. Lowering establishment and maintenance costs for old world bluestem will help producers meet these objectives.

During FY 2001, ARS will

produce recommendations for the best way to provide supplements to calves grazing on bermuda grass in the South Central United States.

ACCOMPLISHMENTS: A finishing system was presented to producers at a field day and articles were published in the "paperless press."

IMPACT/OUTCOME: Producers will have the option to finish calves for market or sell them to feed lots.

ACCOMPLISHMENTS: Stockers of different genetic make-up established that calves from purebred cows grew fatter than calves from crossbred cows. Stocker calves from Brahman cows gained weight faster than stockers from Angus cows.

IMPACT/OUTCOME: Producers can match the genetics for the stockers with their specific system to optimize economic return.

ACCOMPLISHMENTS: Growth curve analysis conducted at STARS indicates that the growth rate of most breeds of calves in the late summer is below their genetic potential. This is in large part due to declining milk production of their dams and low nutritive value of available forage. Researchers at STARS investigated molasses-protein creep supplement as a method for supplying extra nutrients to calves during the late summer. Additionally, another series of experiments looked at creep grazing calves, by providing access to rhizoma perennial peanut, a high quality tropical legume. In this experiment, milk production of the calf dams was actually measured. The molasses treatment did not affect calf weaning weights due to low consumption. Creep grazing did improve weaning weights of calves by about 25 pounds some years, particularly for those dams with low milk production, but this improvement in weaning weight was noted only in years when the calves actively grazed the creep.

IMPACT/OUTCOME: Identified a means that producers in the subtropical United States could use to improve the performance of their calves without using purchased supplement.

STRATEGY 2.1.2: Plant, animal, and ecosystems protection: Improve integrated management systems that contribute to the protection of plants, animals, and ecosystems against pests (insects, weeds, pathogens, etc.).

During FY 2001,ARS will

ACCOMPLISHMENTS: Volatile organic compounds were collected from the skin of humans, fractionated, tested for the attractancy to mosquitoes, and identified. Besides several attractants, five attractancy inhibitors were discovered.

IMPACT/OUTCOME: Both attractants and repellents have value in the control of insect-borne diseases. Attractant inhibitors may lead to new classes of economically competitive, efficacious repellents for use on animals and humans. Attractants can be used to increase the efficiency and specificity of traps used for disease surveillance, as for West Nile virus.

ACCOMPLISHMENTS: In the midsouth United States, the tarnished plant bug, tobacco budworm, bollworm, and cotton aphid have been the most persistent and economically important pests of cotton since the eradication of the bollweevil. Currently, tobacco budworm is being controlled very effectively with transgenic Bt cotton, which is somewhat less effective on bollworm. Control of tarnished plant bug and aphids rely almost exclusively on the use of chemical insecticides. Aphids can be brought under control using the fungus, Neozygites fresenii. ARS scientists at Stoneville and Mississippi State, Mississippi, have recently expanded their efforts for control of the tarnished plant bug using an areawide management strategy. Technologies that are being developed and deployed with varying degrees of success by the scientists include sex pheromones, monitoring for insecticide resistance, destroying potential hosts that supply the tarnished plant bug to cotton, judicious use of insecticides, and release of sterilized tarnished plant bugs into the pests alternate hosts prior to their invasion of cotton fields. The tarnished plant bug so far seems to be a particularly good candidate for use of the sterile insect technology to decrease populations because progeny of partially sterile bugs are also sterile, and two generations or more of sterile bugs has resulted from a single release.

IMPACT/OUTCOME: Implementation of a pheromone trapping and sterile tarnished plant bug release program on an areawide basis, in lieu of insecticide applications, is decreasing populations of the pest below economic levels, as well as giving natural enemies of tarnished plant bug and other cotton insect pests the opportunity to control the pest as well. The program also significantly decreases the environmental impact from the overuse of chemical pesticides.

ACCOMPLISHMENTS: An experimental method for testing selective blue-green algicides in catfish production ponds was developed and used for the past two summers.

IMPACT/OUTCOME: Selective blue-green algicides are being discussed with interested companies for commercialization.

ACCOMPLISHMENTS: Research on a fungal biological control agent for tall whitetop or hoary cress (Cardaria draba) was begun at the Northern Plains Agricultural Research Laboratory in Sidney, Montana. The host-specificity and efficacy of this fungus (Cercospora sp.) is currently being characterized.

IMPACT/OUTCOME: If the Cercospora fungus is specific to whitetop, and damages the plant significantly, it may provide ranchers with a non-chemical, affordable, sustainable option for this weed.

ACCOMPLISHMENTS: Herbicides and conventional management techniques cannot control yellow star thistle, a major Eurasian invasive weed of crops, rangeland and natural areas. ARS scientists with the Foreign Disease-Weed Science Research group at Frederick, Maryland, completed testing and applied for field release of a fungus (Puccinia jaceae). They determined that the fungus was specific and very damaging to yellow star thistle, obtained release approval from Californian regulatory, and are awaiting final Federal release approval.

IMPACT/OUTCOME: This is the first time in the modern regulatory era in the United States that a plant pathogen has gone through the regulatory process, and if the final regulatory hurdles are passed and the fungus is released, there is an excellent chance to reduce yellow star thistle populations that lower rangeland productivity and threaten valuable native plants.

ACCOMPLISHMENTS: ARS had major accomplishments for this indicator for saltcedar and rangeland weeds threatening Indian Reservations.

Saltcedar is one of the most damaging invasive weeds in the western half of the United States, replacing beneficial riparian vegetation, increasing risk to native birds, causing erosion, poisoning livestock, blocking access to water by livestock, drying water sources, and increasing fire hazard and frequency. ARS scientists with the Exotic and Invasive Weeds Research group in Albany, California, and Temple, Texas, released the Chinese leaf beetle (Diorhabda elongata) in six states. They have become established at release sites and are spreading to new infestations of saltcedar.

The Bureau of Land Management estimates that infestation of their lands by invasive weeds nearly quadrupled from 1985 to 2000, and they have no affordable, environmentally friendly methods of managing these weeds. ARS scientists with the Pest Management Research Unit in Sidney, Montana, redistributed more than 10 million biological control agents for leafy spurge, knapweeds, field bindweed, Canada thistle, and poison hemlock to Indian Reservations in Montana. Selected sites on the Reservations will be monitored over the next several years to determine establishment and impact on the target weeds.

IMPACT/OUTCOME: The Chinese leaf beetle (Diorhabda elongata) and other agents that will be released pending completion of host-specificity testing, may provide federal, state and private land managers with effective and affordable methods of control for saltcedar. This significant progress in biological control may provide the only answer for long-term and ecologically acceptable management of the weed.

The technology transfer for these rangeland weeds provides safe, affordable, sustainable and environmentally friendly biological control agents to Indian Reservations. The impact of the agents is reducing temporary, expensive and environmentally damaging inputs to management of the weeds that would otherwise be required. These impacts are expected to increase with time, as populations of agents increase and spread to other infestations of the weeds.

ACCOMPLISHMENTS: Leafy spurge causes more than $150 million damage annually in western states, including loss of rangeland, invasion of natural areas, loss of biological diversity, high herbicide costs, and low profitability. ARS scientists with the Pest Management Research Unit in Sidney, Montana, has coordinated the first areawide integrated weed management project (TEAM Leafy Spurge) in the United States. This project is a partnership between ARS, USDA-APHIS, and universities in Montana, North and South Dakota, and is conducted in cooperation with ranchers, Forest Service, Cooperative States Research, Education and Extension Service, the Bureau of Land Management, National Park Service and the state Departments of Agriculture. All available strategies for leafy spurge management are being investigated.

IMPACT/OUTCOME: TEAM Leafy Spurge has resulted in redistribution of over 20 million Aphthona flea beetles (the main herbivore against leafy spurge) in Montana, North Dakota, South Dakota and Wyoming, and many infestations of the weed are now under control. The roles of chemical, cultural and mechanical control in containment of leafy spurge are being determined.

During FY 2001, ARS will

determine both chronic and acute pain associated with farm management practices (tail docking, beak trimming, induced molting) in conjunction with the expression or lack of expression of behavior response.

ACCOMPLISHMENTS: Research was published and provided to popular press on measurements of pain and expression of behaviors associated with tail docking of dairy cattle.

IMPACT/OUTCOME: This information could impact farm management practices to minimize the pain to animals and improve production.

ACCOMPLISHMENTS: Preliminary data indicated that regrouping pigs versus nongrouping pigs had increased bacteria in the rectum. Tests with more pigs are recommended.

IMPACT/OUTCOME: Eliminating "co-mingling" appears to be a strategy that could reduce food safety risks.

STRATEGY 2.1.3: Germplasm resources and genomics: Acquire, preserve, evaluate, describe, and enhance genetic resources and develop new knowledge and technologies to increase the productive capacity and usefulness of plants, animals, and other organisms.

During FY 2001,ARS will

ACCOMPLISHMENTS: Strawberry germplasm US 4808 and 4809 were released by ARS scientists in Beltsville, Maryland. These are the first and only know sources of resistance to angular leaf spot. US 4808 was collected from a wild population in Minnesota germplasm. US 4809 was collected from a wild population in Georgia.

As part of a program to increase the nutritional value of vegetables, pigmented carrot roots from India, China, Turkey, Syria, and Egypt were intercrossed and visually selected for high color by ARS scientists in Wisconsin. Segregating families from 19 crosses were evaluated chromatographically and high pigment individuals further intercrossed. Further selection and evaluation is underway.

In some crops, complex genetic backgrounds make it difficult or impossible to make crosses with wild relatives. ARS scientists in Madison, Wisconsin, (cucumber) and in Pullman, Washington, (chickpea) are making progress in broadening the genetic diversity of these crops by overcoming crossing barriers with wild relatives.

IMPACT/OUTCOME: Identifying native sources of resistance to angular leaf spot in strawberry and freely distributing this material to genetic improvement programs will allow plant breeders to develop new varieties resistant to this potentially serious disease.

Breeding vegetables such as carrots with new pigments that enhance nutrition is an important step in harnessing genetic diversity in horticultural production. The program at Madison is involved with extensive field trials in key production areas, and is also involved in assessing the impact of improved germplasm on human nutrition, thus the potential impacts of this work on both agriculture and human health are considerable.

In crops with barriers to the introgression of native plant material into breeding populations, efforts such as those with chickpeas (Washington) and cucumber (Wisconsin) are critical to broadening the genetic diversity of important horticultural crops. Pest resistance, crop quality, productivity and adaptability are all potential impacts of broadening the genetic base of these crops.

ACCOMPLISHMENTS: ARS scientists in Salinas, California, (in cooperation with the California Beet Growers Association) released sugarbeet germplasm line M1 which carries a high level of resistance to root-knot nematode. Breeder seed is being maintained at the Agricultural Research Station in Salinas, California, and will be provided to sugarbeet breeders and researchers in small amounts.

IMPACT/OUTCOME: Nematode resistant varieties will be developed using this germplasm, and production potential in nematode infested areas will improve. The profitability of sugarbeet production will increase by increasing sugar yield per acre.

ACCOMPLISHMENTS: ARS scientists at the Southern Regional Research Center developed new methods to identify factors in corn grain that impede aflatoxin formation. They have developed tests to identify protein inhibitors of aflatoxin production. As a first step, corn lines were identified that demonstrate significant inhibition of toxin production without affecting fungal growth.

IMPACT/OUTCOME: Identification of corn lines that inhibit toxin production will enable the ARS scientists in New Orleans to now identify specific corn proteins that confer natural resistance to aflatoxin production following fungal invasion. This research may provide a new breeding strategy to enhance natural antifungal resistant mechanisms in commercial corn hybrids.

ACCOMPLISHMENTS: In addition to having successfully genetically engineered papaya against Papaya Ringspot Virus (PRSV), a devastating disease for production in Hawaii, ARS scientists are now making crosses to transfer this critical trait into a number of commercially important varieties and are making progress in increasing fruit size and quality.

ARS scientists at Beltsville, Maryland, had genetically engineered tomato fruit to have longer vine and shelf life, robustness, and a 3-fold increase in the lycopene content.

Transformation systems are being developed for a number of other crops to improve crops such as blackberry for cold hardiness (Corvallis, Oregon), pineapple for nematode resistance (Hilo, Hawaii), peaches for plum pox virus resistance (Kearneysville, West Virginia), citrus for pest resistance and horticultural traits (Fort Pierce, Florida) and walnuts for control of aflatoxin (Albany, California).

IMPACT/OUTCOME: Many horticultural crops (such as pineapple, peaches, citrus) are threatened by devastating diseases which would reduce fresh fruits and vegetables availability to the consumer. Papaya was threatened by a devastating disease but through genetic engineering resistant plants are now producing in areas otherwise not productive due to a serious virus problem. Increasing quality and nutritional characteristics of tomato will increase the quantity, quality and nutritional value of tomato products available to the consumer. Oregon is the major national producer of blackberries but that crop is often devastated by cold weather, thus a cold hardy blackberry would increase stability of production and supply while making this nutritious fruit more available to consumers. Potential food safety hazards such as aflatoxin in nut crops like walnut can be addressed with biotechnology in order to insure the safety of the food supply for the consuming public.

ACCOMPLISHMENTS: Sixty-three collections of white clover were made from well managed pastures in Kentucky, Ohio, and West Virginia. The collection was established and evaluated at the USDA-NRCS plant material center at Alderson, West Virginia. Many of the collection proved superior in terms of production and persistence and the five best populations were selected to grow out seed to make this germplasm available to plant breeders.

IMPACT/OUTCOME: Because these clover populations are well adapted to the region, they should be an excellent source of germplasm to develop high-quality forage varieties well suited for the harsh growing conditions of Appalachia and reduce input costs for limited resource farmers.

ACCOMPLISHMENTS: Root cultures of trefoil (Lotus corniculatus) were evaluated as a screening tool for evaluating germplasm for desirable traits. None of the tests evaluated proved more economical than conventional screens. However, in screening for resistance to soybean cyst nematodes (SCN), scientists at Columbia, Missouri, observed that the presence of trefoil roots caused SCN eggs to germinate but the juveniles that did invade the roots failed to lay eggs.

IMPACT/OUTCOME: These preliminary results indicate that using a trefoil crop in rotation with soybeans may be good biological control for reducing SCN in heavily infected fields.

ACCOMPLISHMENTS: Plant explorations coordinated and supported by USDA/ARS successfully collected germplasm in nine nations, including the United States. Samples of pears, breadfruit, tomato, forages, spinach, onion relatives, chiles, amaranths, and lesquerella were secured and added to the National Plant Germplasm System (NPGS).

IMPACT/OUTCOME: These explorations safeguarded a wide diversity of genetic material in NPGS genebanks, where they will available on request to researcher and breeders worldwide. Access to diverse genetic material is key to ensuring continual progress in crop genetic improvement; the latter is integral to food security for the ever increasing human population.

ACCOMPLISHMENTS: USDA/ARS scientists released varieties from a wide spectrum of crops, including disease-resistant broccoli, "soyfood" cultivars for U.S. production and export to Japan, new dwarf cultivar of African iris for the nursery trade in southern Florida, a new longer-lived and more productive hybrid citrus rootstock, a new rice variety for the southern United States with unique, highly effective resistance to the serious diseases and also with superior yield potential and grain quality, several superior potato varieties, 14 pinto bean lines (of various market classes) resistant to common bacterial blight, two new varieties of dry pea resistant to many diseases, and with improved seed quality, new lesquerella (a specialty oilseed) varieties with superior agronomic traits, and the first sugarbeet line resistant to root knot nematode.

IMPACT/OUTCOME: Some of the new crop lines released by USDA/ARS scientists provided very high levels of resistance to diseases and pests, thereby serving as sources of resistance genes for further breeding. Rice cultivars with higher yield, better grain quality, and superior resistance to diseases and insects may increase and sustain profitability for U.S. rice producers.. New ornamental varieties increase the diversity of products for U.S. producers, potentially increasing profitability. Other new varieties will enable U.S. farmers to remain competitive in the global export market, e.g., the new citrus rootstocks released potentially may save Florida citrus producers millions of dollars each year, the new lesquerella varieties should lower production costs. Finally, new disease resistant and more productive varieties (e.g., pea, potato) should increase producer profitability.

During FY 2001, ARS will

ACCOMPLISHMENTS: An article is in press that describes the identification and sequencing of three different transcripts associated with disease resistance.

IMPACT/OUTCOME: This research is the basis for developing molecular markers with potential for developing genetic lines of catfish with strong innate immune functions. Innate immune functions are believed to be critical for resistance traits against causative agents such as ESC.

ACCOMPLISHMENTS: At the end of 2000, an international scientific team that includes ARS researchers in Albany, California, finished the first essentially complete genome sequence for a plant, Arabidopsis, an experimental "model plant." It was sequenced first because of its small genome and rapid life cycle.

IMPACT/OUTCOME: Knowledge of the Arabidopsis genome is already helping to elucidate the structure and function of crop genomes and their constituent genes. For example, scientists at Albany, California, are currently using this information to find genes in maize that govern agronomically-important traits.

ACCOMPLISHMENTS: At the end of 2000, an international scientific team that includes ARS researchers in Albany, California, finished the first essentially complete genome sequence for a plant, Arabidopsis, an experimental "model plant." It was sequenced first because of its small genome and rapid life cycle.

IMPACT/OUTCOME: Knowledge of the Arabidopsis genome is already helping to elucidate the structure and function of crop genomes and their constituent genes. For example, scientists at Albany, California, are currently using this information to find genes in maize that govern agronomically-important traits.

ACCOMPLISHMENTS: With the USDA/ARS Center for Agricultural Bioinformatics at Cornell University, Ithaca, New York, serving as a hub, the USDA/ARS effort in bioinformatics and crop genome databases expanded its overall capacity. USDA/ARS scientists and collaborators at the Center tested a prototype "data warehouse" for managing millions of individual cross-sequence comparisons, and assessed the utility of software components that may be assembled into a high-volume, high-speed sequence analysis "data pipeline." USDA/ARS scientists at Columbia, Missouri, and their university collaborators integrated data in the database MaizeDB so that the preceding sources of information are linked, and so that MaizeDB is more interoperable with other genome databases. USDA/ARS scientists at Albany, Calfornia, produced more than 70,000 individual ESTs (expressed sequence tags, unique, small identification sequences for individual genes) for wheat, barley and their close wild relatives, which is more than 85 percent of such ESTs that are publicly available worldwide. USDA/ARS scientists at Hilo, Hawaii, and collaborators used more highly refined genetic maps of the related crop sorghum?whose genome is much smaller and simpler?as templates for constructing a superior genetic map for the two important sugarcane species.

IMPACT/OUTCOME: The efforts at the Center at Cornell University and elsewhere will enhance the USDA/ARS genome database and bioinformatic capabilities to effectively manage and deliver to users the masses of sequencing data issuing from the sequencing of model species genomes, and of ESTs. Improvements to the maize database that integrate information about gene sequences, gene function, and chromosomal location may facilitate the isolation and analyses of genes controlling important agronomic traits in maize. The process of identifying individual genes by unique tags (ESTs) is essential for further genome analyses, and will furnish a highly valuable tool for accelerating the progress of cereal genetics and breeding. The mapping information derived from the sorghum genome may help accelerate genetic research and crop improvement in sugarcane, via marker assisted selection.

ACCOMPLISHMENTS: Transformation technology was developed for several crops that allows creation of genetically engineered plants for specific purposes. Barley, potato, and the "queen of ornamentals," the rose, are conspicuous examples. Transformation of cotton has also been advanced to the point that it can be reliably used to create novel genetic combinations.

IMPACT/OUTCOME: Under a cooperative agreement, transformation technology for roses was advanced to a useful state, and the technology was transferred to the private sector partner for use in creating disease-resistant roses.

Barley transformation technology has been considerably advanced, and is now useful for generating disease-resistant genetic lines. Testing is underway for genes that may provide resistance to head scab (Fusariumgraminearum), and genes for powdery mildew resistance will be tested in the future.

Under cooperative agreements, transgenic potatoes have been developed that reduce glycoalkaloid content of the tuber and increase late blight resistance.

Cotton has been transformed with a haloperoxidase gene that generates low concentrations of the active ingredient of laundry bleach, hypochlorite. The transformed plants are undergoing testing for resistance against Aspergillus flavus, the pathogen that synthesizes aflatoxins.

During FY 2001, ARS will

ACCOMPLISHMENTS: Scientists at Beltsville, Maryland, have successfully developed protocols for genetically engineering highbush blueberry. Other scientists within the group are studying other traits such as the molecular basis of cold hardiness, disease resistance, and fruit quality. In concert with the ARS blueberry breeding program at Chatsworth, these technologies will be useful in developing research models to understand these systems, and in the future may be used to produce varieties with characteristics not attainable through conventional breeding.

ARS sugarcane breeders at Houma, Louisiana, and Canal Point, Florida, have used new approaches in molecular genetics to enhance the conventional breeding program. In addition to targeting critical traits in pest resistance and production efficiency, marker assisted breeding strategies have been developed which will greatly enhance the highly labor and time intensive breeding protocols involved in developing new sugarcane varieties. Molecular markers identifying wild parents will help breeders to eliminate crosses in which self pollination has occurred, greatly enhancing selection efficiency.

ARS sugarbeet scientists at Ft. Collins, Colorado, breeding for multiple disease resistance have combined traditional and molecular technologies to produce germplasm more rapidly and efficiently to meet the changing needs of seed companies and the growers they serve.

ARS lettuce breeders in Salinas, California, are developing techniques for marker-assisted selection for the enhancement of germplasm for a number of traits, including resistance to critical diseases such as lettuce mosaic and big vein. Sources of resistance have been successfully identified and in some cases the improved germplasms could actually be used as cultivated varieties if that were necessary.

ARS scientists at Madison, Wisconsin, have identified molecular markers for several genes of the carotenoid and anthocyanin biosynthetic pathways and have developed carrot germplasm with high pigment content and sweet, mild flavor. In addition to the field testing of improved germplasm for production potential, human clinical trials are being undertaken for bioavailability of key phytonutrients.

IMPACT/OUTCOME: Developing marker assisted strategies in conventional breeding programs of horticultural crops have tremendous potential to increase the efficiency and effectiveness of long term, resource intensive breeding programs. By more effectively targeting breeding populations, plant breeders can more quickly provide disease resistant germplasm and varieties which will increase production efficiency and thus increase availability of produce to the consumer, sustainability for the producer and will conserve land and natural resources while reducing the use of crop protectants. In addition, marker assisted selection will increase the ability of breeders to target important quality traits that will increase crop utilization and enhance human health.

ACCOMPLISHMENTS: Tobacco and cotton plants (the former as a model species for testing purposes) were genetically engineered to express a bacterial gene coding for chloroperoxidase (which generates compounds related to household bleach), and numerous genetic lines resulting from this gene introduction were evaluated. Extracts from four of the cotton plants were active against cultures of the aflatoxin-producing fungus, Aspergillus flavus. When seeds were removed from the plants and inoculated with the fungus, they were resistant to infection. Engineered tobacco plants also showed a hypersensitive response against some other fungal disease organisms. A patent is pending for this use of the chloroperoxidase gene (the gene itself is not proprietary).

IMPACT/OUTCOME:The cotton breeding pool contains little useful genetic resistance to aflatoxin-producing fungi. Genetically engineered resistance is expected to greatly enhance the ability of breeders to develop cotton varieties with reduced aflatoxin problems.

ACCOMPLISHMENTS: ARS, in cooperation with North Dakota State University, has developed a cultivar named HiFi, which has 50 percent more beta-glucan than any other oat cultivars. This cultivar is undergoing final evaluation prior to release.

IMPACT/OUTCOME: The soluble fiber, or beta-glucan in oats has been shown to reduce serum cholesterol in humans and actually decreases the risk of heart disease when incorporated into the diet. The assimilation of grain from this cultivar into the national food supply may more greatly enhance health benefits currently enjoyed from oat consumption.

ACCOMPLISHMENTS: ARS, in collaboration with North Dakota State University, tested a waxy (low amylose) durum wheat for bread baking quality. Results indicate a substitution of fat with 20 percent waxy wheat flour results in a loaf of bread with softness and shelf life improved over the no fat control and just as good as the 3 percent shortening control.

IMPACT/OUTCOME: Development of a waxy durum wheat with good bread baking quality indicates that a waxy durum wheat variety could be used to produce a no fat bread with improved softness and shelf life.

ACCOMPLISHMENTS: Two varieties of red clover and one variety of kura clover are being released this year and pre-breeder seed is available for a new variety of birdsfoot trefoil.

IMPACT/OUTCOME: The red clover varieties are disease resistant, higher yielding, and persist longer(3-4 years). As a result, they will save on average over a $100 per acre in re-establishment and maintenance costs per year. The trefoil and kura clover are also more disease resistant and persistent.

ACCOMPLISHMENTS: P-7 bluebunch wheatgrass germplasm was released as a selected class of certified seed (genetically manipulated track).P-7 is a multiple-origin ploycross produced by intermating 25 different populations.

IMPACT/OUTCOME: Bluebunch wheatgrass is a native grass species found in many parts of the West and is an important species for ecosystem restoration projects following wildfires and the control of invasive weeds. P-7 is intended to provide a high level of genetic diversity within a single release to aid in maintaining genetic biodiversity during ecosystem restoration.

ACCOMPLISHMENTS: At Lincoln, Nebraska, populations of switchgrass germplasm synthesized from plant materials collected from the tallgrass prairiehave been advanced through two generations of random mating. Seed for this cultivar has been increased and will be released in 2002. A new cultivar of Canada wildrye will also be released in 2002.

IMPACT/OUTCOME: These cross-pollinated cultivars of native grasses will be useful for re-vegetating land to restore native prairies and help preserve genetic biodiversity within the species.

ACCOMPLISHMENTS: The white clover research unit at Mississippi State was re-directed into animal waste research in 1999 and only one new clover cultivar was released prior to redirection.

IMPACT/OUTCOME: The new cultivar has increased disease resistance and this will increase persistence and productivity.

ACCOMPLISHMENTS: Significant progress has been made although drought in Georgia has slowed the release of these new varieties. Three new pearl millet varieties have been developed (99A1, Tift 454,and Tifgrain 102) and in 2002, over 1,000 acres will be planted to provide seed in quantity and to further verify adaptability to local conditions. Potential parents for improved seeded bermuda grass have been evaluated and crossed. In 2002, sufficient seed will be produced to permit field evaluation.

IMPACT/OUTCOME: Southeastern poultry producers need these pearl millet varieties so they can locally produce grain with no preharvest aflatoxins with minimal irrigation inputs while eliminating transportation costs resulting from importing grain from other regions. Existing hybrid bermuda grasses for the southeast cannot be established from seed; they must be sprigged. An improved seeded bermuda grass will help limited resource farmers improve their pastures at a more affordable cost.

During FY 2001, ARS will

ACCOMPLISHMENTS: ARS, in cooperation with peanut breeders in Florida and North Carolina, determined trends in peanut flavor quality in all varieties released since 1930. Virginia-type cultivars showed an overall trend toward reduced flavor quality in recent varieties. Considerable variation in roasting quality was found among runner-type cultivars released since 1980. Spanish and fastigiate varieties exhibited consistent improvement in sweet and bitter components of roasted peanut flavor intensity.

IMPACT/OUTCOME: These data established that specific ancestral cultivars contributed characteristics that reduce the flavor quality of some new peanut breeding lines. However, this new knowledge has enabled the development of genetic methodology to select future parental lines and cultivars with superior flavor profiles.

ACCOMPLISHMENTS: A new formula was implemented for net genetic merit in the National Dairy Cattle Genetic Evaluation Program.

IMPACT/OUTCOME: The implementation of the new formula increased the effectiveness of the selection and raised the emphasis on traits selected to meet industry goals.

ACCOMPLISHMENTS: Calving ease was added in the National Cattle Genetic Evaluation Program.

IMPACT/OUTCOME: Dairy producers now have information available to them concerning the identification of "easy calving bulls" to breed to heifers, and how to use these bulls to reduce the problem of difficult calving.

ACCOMPLISHMENTS: In the first phase of this project, cytogenetic studies of the 4 nondormant alfalfa germplasm sources were completed using chromosome banding and image analysis techniques. The four sources could not be distinguished from each other.

IMPACT/OUTCOME: These results indicate that the germplasm had not been derived from original sources and/or germplasm from several sources had been mixed.

ACCOMPLISHMENTS: A microsatellite-based genetic linkage map for channel catfish was published in June 2001.

IMPACT/OUTCOME: Use of microsatellite and genetic linkage maps will increase the efficiency of selective breeding programs for channel catfish and rainbow trout.

STRATEGY 2.1.4: Plant and animal biological processes: Develop biologically-based technologies to improve productivity, safety, nutrient content, and quality of plants, animals, microbial organisms, and their products.

During FY 2001, ARS will

ACCOMPLISHMENTS: An environmental assessment of the effect of aquaculture use of copper sulfate was submitted to the Federal Drug Administration (FDA). A critique of the submission has been received from FDA and a revision of the assessment has been submitted.

IMPACT/OUTCOME: This assessment allows FDA to determine whether the benefit of the use of copper sulfate to control diseases in aquaculture warrants the risk it poses to the environment.

ACCOMPLISHMENTS: Efficacy studies of potassium permanganate for preventing the establishment of Ichthyophthirius multifiliis was completed.

IMPACT/OUTCOME: This research will provide the basis for FDA to approve a label claim for potassium permanganate.

ACCOMPLISHMENTS: A U.S. patent was granted for a modified, killed Streptococcus iniae vaccine for the prevention of streptococcal disease in tilapia and hybrid striped bass. An application was submitted to the U.S. patent office for a Flexibacter columnaris vaccine.

IMPACT/OUTCOME: A cooperative research and development partner (Intermet) has been identified to commercialize their vaccine.

ACCOMPLISHMENTS: Research was completed that showed variation for natural resistance to columaris disease among strains of catfish.

IMPACT/OUTCOME: Differences among catfish were not at a magnitude to justify additional research.

ACCOMPLISHMENTS: ARS scientists at Plum Island, New York, Athens, Georgia, Laramie, Wyoming, Beltsville and Frederick, Maryland, and Ames, Iowa, working in collaboration with the Department of Defense, have developed rapid on-site tests that detect and identify important animal, plant, and foodborne pathogens.

IMPACT/OUTCOME: Development of these new diagnostic rapid detection technologies allow animal health officials in regulatory capacities (APHIS and State agencies) greater ability to determine if a disease agent is present, where it is located, and when it is eradicated (if possible). This will aid in ensuring our trading partners have confidence in our ability to detect and control disease agents.

OBJECTIVE 2.2: Safe food: "Maintain an adequate, nutritious, and safe supply of food to meet human nutritional needs and requirements."

STRATEGY 2.2.1: Plant and animal product safety: Provide knowledge and means for production, storage, and processing of safe plant and animal products.

During FY 2001, ARS will

ACCOMPLISHMENTS: ARS has undertaken several field trials of the competitive exclusion (CE) product for swine. These trials will be used to help meet the FDA requirements for the approval for use of this product to help control Salmonella and E coli.

IMPACT/OUTCOME: Because the product also improves swine production, extensive use by swine producers is expected, and the product has the potential to significantly help assure safe meat products.

ACCOMPLISHMENTS: ARS developed rapid immunoassays for the detection of various Campylobacter species, Listeria, including specifically Listeria monocytogenes, and E. coli, including O157:H7 and other Enterohemorrhagic Escherichia coli strains. The assays for detection of antibiotic residues was only completed in part due to realignment of the food safety program to address the more critical issues of the detection of molecules associated with the transmission of the bovine spongiform encephalitis infective agent; e.g., bovine proteins, central nervous system tissues, and prions in animal feed. The scientist involved in developing immunoassays for antibiotics was transferred from the College Station, Texas, laboratory to the Western Regional Research Center, Albany, California. He will continue, albeit at a slower pace, to develop immunoassays for antibiotic residues since this is still a major concern for the Food Safety Inspection Service (FSIS). It is anticipated the residue assays will be available by 2005.

IMPACT/OUTCOME: The immunoassay technologies that were completed were transferred to FSIS to assist the Agency in fulfilling its regulatory requirements. Various industry partners, including Igen Corporation based in Maryland, are associated with the technology transfer for the food industry.

ACCOMPLISHMENTS: Studiesindicated that the GRAS status herbal extract did not produce the required 5 log (99.999 percent) reduction in Salmonella levels required by regulatory agencies for use by industry.

IMPACT/OUTCOME: The project scientists were redirected to another poultry processing problem.

ACCOMPLISHMENTS: ARS has demonstrated that the bacterial endophytic strain of Bacillus subtilis can control plant diseases and mycotoxins caused by certain fungi.

IMPACT/OUTCOME: The use of this bacterium, which has been patented,will provide the potential for substantive control of fungal growth to help assure safer corn and other food commodities.

ACCOMPLISHMENTS: ARS scientists at Raleigh, North Carolina, determined the degree of experimental error associated with sampling, sample preparation, and analysis of mycotoxin concentration in corn. These statistical measures enabled more accurate estimates of aflatoxin and fumonisin in shelled corn.

IMPACT/OUTCOME: An improved method was designed and evaluated for determining the performance of sampling plants for analysis of aflatoxin fumonisin content in shelled corn.

ACCOMPLISHMENTS: Under a CRADA with Dow AgroSciences, tobacco and cotton plants were engineered to express a synthetic gene coding for a lytic peptide, resulting in enhanced disease resistance. The demonstration showed the potential of the proprietary synthetic peptide for disease prevention. The CRADA ended in 2001 and the information has been transferred to the private sector partner.

IMPACT/OUTCOME:The synthetic peptide was shown to enhance resistance of cotton to the important toxin-producing pathogen, Aspergillus flavus. This engineered trait, when incorporated into cotton varieties, will be the first source of significant genetic resistance to this troublesome pathogen.

ACCOMPLISHMENTS: Genes identified with resistance to aflatoxin have been incorporated into the cotton genome and plants grown in the laboratory. The next step is to produce cotton seed to determine if the trail is stable and produces acceptable levels of resistance.

IMPACT/OUTCOME: If successful, these engineered plants will be resistant to the production of aflatoxin, thus greatly reducing this serious food safety and economic threat to cotton and other commodities.

ACCOMPLISHMENTS: ARS scientists traced and enumerated E. coli O157:H7 in many cattle production operations. They also followed these animals to slaughter and determined the number of carcasses carrying the pathogen. The number of pathogens was consistently higher in the summer than in the winter months, but there was no consistent geographic trend. ARS scientists have also determined the presence of this pathogen in water and in several vectors in the production environment, as well as measuring the effects of different feeding regimes. Additional studies will be necessary before definitive control procedures can be recommended.

IMPACT/OUTCOME: The information gained in these studies has the potential to help significantly reduce the levels of E. coli O157:H7 in cattle and thus in beef products.

ACCOMPLISHMENTS: Combination treatment processes for the microbial decontamination of pork and beef trim were designed and evaluated, for use at the last possible step prior ton grinding. These treatment processes were shown to reduce and control populations of fecal bacteria on beef or pork trim and in the resultant ground product without a large negative effect on ground meat quality. A method was also developed to rinse sample beef trim pieces for the presence of E. coli O157 and other pathogens. Trim intervention technology will have to first be adapted by engineering development for optimal delivery to all surfaces of meat trim in a manner that is economically feasible, environmentally safe (with regard to increased effluents) and, most importantly, will not cause quality or functionality problems in the resulting products.

IMPACT/OUTCOME: Information on various accomplishments related to antimicrobial interventions to reduce both pathogenic and spoilage bacteria on carcasses and red meat has been presented to scientific and industrial groups, livestock breeders and producers, and other professionals that are potential end-users of the science and/or technologies to decrease the likelihood of pathogens entering the food chain.

develop techniques to improve the effectiveness of washing technologies for pathogen decontamination of fresh fruits, vegetables and seeds. Determine if ionizing irradiation or a combination of it and subsequent washing with a sanitizer is more efficacious. Develop a profile of the environmental conditions required to minimize growth of pathogens on fruits and vegetables during storage.

ACCOMPLISHMENTS: Current washing and sanitizing methods used by industry, even with small modifications, have proven ineffective or unworkable for the most part, to achieve the new 99.999 percent (5 log) reduction in pathogens required by the FDA. While the research program is still in progress, there has however, been some success. A method to decontaminate sprouting seed while maintaining quality and viability was designed using a combination of gamma irradiation and chlorine. Promising multiple hurdle treatments using combinations of physical (brushing) and chemical (hydrogen peroxide and citrus-related antimicrobial compounds) treatments have been developed for such products as cantaloupes, apples, peppers, potatoes, and lettuce. While these techniques work in the laboratory, they need to be evaluated in industrial processing conditions. In association with Penn State University, ARS, at the Eastern Regional Research Center, has built a specialized Biological Safety Containment Facility (BSL-2) for use in evaluating new washing technologies and for investigating effects of commercial handling, packing, and processing practices. It is envisaged that studies will be completed in 2004. Studies on developing environmental conditions to reduce pathogen growth continue. The numbers and types of pathogens on produce and how they interact with tissue and each other to form stable biofilms has proven to be challenging. Developing alternate conditions has also been complicated by the now required use of lower levels of GRAS chemicals. Alternate methods using natural competitive exclusion cultures are being evaluated.

IMPACT/OUTCOME: Information on techniques for decontaminating produce and sprouting seed have been disseminated to growers, processors, trade organizations, and Federal regulatory agencies. The method to decontaminate sprouting seed was adopted by the FDA, approved by the sprout industry, and received permanent registration by the EPA. This method has had a significant impact in reducing the incidence of sprout related food-borne illness. It is anticipated that technologies developed for other produce will have a similar positive effect when implemented.

ACCOMPLISHMENTS: Completion of the project was hindered by the retirement and transfer of scientists from within the project. Consequently, the project was combined into a larger program, the National Animal Health Monitoring System (NAHMS), to collect information to predict trends in food-borne carrier rates, antibiotic usage and resistance, and biosecurity.

IMPACT/OUTCOME: What data was available from the swine model processing plant study was transferred to the FSIS for incorporation in their HACCP program. The swine processing plant is now part of the Health Improvement and Modernization Programme (HIMP), a larger Hazard Analysis program using "Gold Standard" processing plants. The information gained from the NAHMS program will be used by both industry and regulatory agencies to improve animal husbandry practices to contribute to overall food safety and quality by reducing or eliminating pathogens.

ACCOMPLISHMENTS: ARS has provided and continues to provide data to various regulatory agencies in order to develop risk assessment models. Risk Models for Salmonella enteritidis, E. coli O157:H7, and Listeria have been developed and are currently under review. A model for Campylobacter has been developed but is not publically available. The complexity of food matrices and the lack of data still complicates the implementation of the assessments. ARS continues research to specially focus on Listeria in ready-to-eat foods and E. coli O157:H7 in ground beef. A new series of predictive and simulation models (Poultry-FARM) have been developed that assess the severity and risk of Salmonella and Campylobacter infections from chicken.

ACCOMPLISHMENTS: A new version 6.0 of the Pathogen Modeling Program (PMP) has been developed which estimates the effects of multiple variables on the growth, inactivation, and survival of food-borne pathogens. Specifically growth, nonthermal inactivation/survival, thermal, gamma-irradiation, cooling-growth, time-to-toxigeneis, and time-to-turbidity models were developed.

ACCOMPLISHMENTS: Various innovative methods (low-pressure gas chromatographic/mass spectrometry, super-critical fluid extraction, accelerated solvent extraction, pressurized water extraction combined with solid-phase microfiber, immunoassays) were developed and optimized to substantially speed-up and increase the detection limits for analysis of hormones, residues, and antibiotics in foods, while concomitantly reducing, if not eliminating, the need for organic solvents in the extraction process.

IMPACT/OUTCOME: The technologies have been adopted by numerous industrial organizations and State and Federal regulatory laboratories in order for them to assure the safety of foods. The SFE methods have been verified and have passed the Association of Analytical Communities International Inter-laboratory Collaborative Study Process, meaning they are the gold standard international methods. Our technologies have been adopted by industry CRADA partners. Immunoassays and the monoclonal antibody cell line for the beta-agonist ractopmine were issued a U.S. patent.