DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration

Memorandum

One of the goals of the Food Safety Initiative (FSI) is to optimize the use of the limited resources available for Federally supported food safety research. On an interagency basis, this goal is being realized through increased coordination of our research programs with those of our sister agencies. This effort includes direct interaction and collaboration with these agencies and support of the formation of the Joint Institute for Food Safety Research (JIFSR).

As part of this process, CFSAN has been working to optimize intramural FSI research programs. The focus has been to identify priority research needs and devote sufficient resources to a limited number of research projects to realize these priority needs in a timely manner. Integral to the management of this initiative is a strong emphasis on program accountability and "mission relevance." The development of the CFSAN 3-Year Research Plan during FY98 was the Center's first step in this effort. One of the goals of that document, as a research management tool, was to communicate CFSAN's priority research needs, as identified in the FSI and PIFSI 1997 reports and by the managers of CFSAN's regulatory programs.

During the past six months, the FSI Research/Risk Assessment staff has been working with the Center's research scientists and regulatory program managers to improve the means for identifying the Center's priority research needs and communicating those needs to: (1) FDA scientists; (2) other Federal research agencies; and (3) our industry and academic research partners. Within CFSAN, the goal is to provide the research scientists, their line managers, and project administrative liaisons with information needed to modify current research plans as part of yearly updates of the 3-Year Plan. Likewise, this information is critical to the development of new research projects, as the current projects reach the point where they need to be revised, redirected, or replaced. The identified priority research needs also serve as a key selection criterion for the competitive intramural research programs that will be announced shortly.

Attached are the two documents that have resulted from our discussions with the regulatory program managers. The first is a letter to the USDA research agencies identifying our research needs that we feel they may be better able to realize because either they have unique research facilities available or we have insufficient resources to adequately address the problem. Each of the areas identified in that document, however, can be considered a priority research need by FDA scientists, particularly if we have unique capabilities for addressing the problem area. The second document is the most recent consolidation of the 3 to 5 year research needs of CFSAN's regulatory programs. The research needs are organized by commodity groups, reflecting the organization of the Center. Thus, it is not surprising that there is overlap between several of the areas.

Together, the two attached documents provide CFSAN's priority research needs for the next 3 to 5 years as identified by the managers of our regulatory programs. It should be noted that these needs are provided in the form of "risk management questions." In so doing, the goal is to provide CFSAN scientists with clear guidance on the "deliverables" that are needed, while empowering you to use your creativity to determine how to best meet these needs in a timely manner. Both documents are provided as a tool to help CFSAN research scientists ensure that their research is directed to priority, mission-relevant needs. Research that addresses either the regulatory needs identified in these documents or those identified in the FY-1998 3-Year Research Plan will be considered priority research for the Center.

Any FSI-supported research scientist whose current research activities do not address one of these needs should discuss, as soon as possible, their research plans with their supervisor and the administrative liaison of the FSI project to which they are currently assigned. It is important to note that virtually all research activities underway currently by FSI-funded research scientists address one or more of these regulatory research needs. Thus, any shifts in current research activities to address a newly identified research need should be done in consultation with your division director and the administrative liaison of your current project. Any substantive changes in projects should be communicated to the FSI research/risk assessment staff so that we can continue to help the office directors ensure that CFSAN has a balanced research FSI program that avoids unplanned redundancies. We, in turn, will be working with the division directors to provide further guidance of relative priorities within each of the research categories.

Finally, it is realized that the scope of the research needs outlined in these documents is well beyond the limited capabilities of the CFSAN intramural research program to address fully. Thus, these documents will also be made available to the public, with the hopes that researchers in academia, industry, and other research agencies will also be interested in addressing some of the questions posed.

Attachments, 2

cc: CFSAN Management Team
CFSAN Office Directors
Dr. William Raub, Assistant Secretary, DHHS, OSASPE
Dr. Steven Sundlof, Director, CVM
Dr. Norris Alderson, Science Director, CVM
Dr. Carl Cerniglia, Director of Microbiology, NCTR
Dr. Bernard Schwetz, Director, NCTR
Dr. James Yager, Director, ORA

RESEARCH NEEDED TO SUPPORT FDA/CFSAN
REGULATORY PROGRAMS
FY-1999

FRESH/FRESH CUT PRODUCE AND RELATED COMMODITIES

PRODUCE, GENERAL:

The priority research areas reflect CFSAN's need to (1) provide the field investigators and laboratories with more effective and cost-efficient means for analyzing produce samples for various pathogenic microorganisms, (2) develop guidance for industry on "good agricultural practices" and "good manufacturing practices" that will help assure the safety of fresh and fresh-cut produce, and (3) identify and evaluate the effectiveness of strategies and technologies that can prevent, reduce, or effectively eliminate pathogenic microorganisms on produce.

• What is the incidence and prevalence of foodborne pathogens (baseline information) on fruits and vegetables associated with outbreaks of foodborne illness (e.g., berries, cantaloupes, lettuce)? Are there any indicator organisms, quality attributes, or other product characteristics that can be used by industry or regulatory inspectors to predict an increased risk of contamination?
• For which pathogens and commodities is infiltration of pathogens into plant tissue likely? What conditions and process foster the infiltration of pathogens? How effective are intervention technologies at reducing the level of internalized pathogens in various fruits and vegetables?
• What rapid techniques are available to reliably determine the level of viable Cryptosporidium parvum oocysts on produce?
• Does hand sorting of produce lead to an increased risk of cross contamination? Does the use of disposable gloves reduce the potential for cross contamination? By how much? How effective are solutions in the hand-dip stations in eliminating surface pathogens from hand or gloves? Is there a minimal time required for dipping to effectively eliminate surface pathogens? Are the industry procedures/standards adequate?
• What levels of microbial reduction (e.g. number of log cycles) are achievable through the use of the following surface treatments at different points in packinghouse operations?

• What is the mechanism of action of different chemical sanitizers on microbial cells? What factors influence the lethality of these sanitizers to various pathogens? What is the potential for development of increased resistance to these antimicrobial agents?

• In the development of performance criteria for the inactivation of pathogens on produce, can the effects obtained with the use of multiple or sequential use of chemical sanitizers be considered additive? Are there instances where agents are synergistic. Should the effects of chemical sanitizers in combination with other interventions technologies (e.g., heat processing, high hydrostatic pressure, ionizing irradiation) be considered additive? Synergistic?
• Should a clean water rinse(s) be required before a chlorine solution or other sanitizing solution is used to treat produce? Should a clean water rinse(s) be required after treatment with a sanitizing solution for produce that is consumed raw?
• What reaction byproducts or residues remain in or on produce from treatment with different chemical sanitizers, especially those that have yet to receive premarket approval for use with produce?

• What is the depth of penetration into different commodities by different chemical sanitizers?
• Does the use of treatments with sanitizing agents eliminate the non-pathogenic microflora such that the commodity would be more likely to support the growth of pathogenic bacteria if contaminated subsequently?
• How do environmental parameters (e.g., temperature, pH, salt concentration, water activity, oxygen tension) affect the inactivation kinetics of foodborne pathogens by different intervention technologies?
• What dose levels of ionizing radiation can be used on commodities of interest (e.g., berries) to eliminate pathogenic microorganisms but maintain quality attributes (e.g., taste, texture)? What are the other relevant treatment parameters in addition to dose?

• Have high hydrostatic pressure techniques been tried on any of the commodities of interest (i.e., those associated with outbreaks of foodborne illness? What are the effects of this potential intervention on sensory attributes? What microbial reductions can be achieved? What are the relevant treatment parameters that influence the efficacy and practicality of the process (i.e., achieve a significant microbial reduction while maintaining a product that is acceptable to consumers)?
• Can competitive exclusion techniques be used to control or reduce the incidence of foodborne pathogens on fresh or fresh-cut produce? What is the efficacy of such techniques? What criteria should be used to evaluate the safety and efficacy of competitive exclusion cultures?
• What packing environments (e.g., oxygen tension, pH, water activity) have the potential to establish conditions that favor growth and toxin production by anaerobic and facultatively anaerobic pathogens?
• What is the effect of MAP and VAC packing of fresh and fresh-cut produce on outgrowth of pathogens (including spore-formers) vs. non-pathogenic, competitive microorganisms in untreated products? In products that have been irradiated or treated with surface sanitizing agents?

SPROUTS:

The research needs related to sprouted seeds fall into the general areas associated with our ability (1) to provide guidance to both our inspectors and industry on how sprouts can be produced safely, (2) to evaluate for premarket approval the plausibility and safety of proposed approaches for the prevention, reduction, or effective elimination of pathogenic bacteria on seeds or sprouts, and (3) to assess the likelihood that sprouts have been contaminated with foodborne pathogens.

• What is the incidence and prevalence of foodborne pathogens on seeds used to produce sprouted seed products? On the sprouted products themselves? How do these compare with seeds associated with sprouted seeds implicated in outbreaks of foodborne illness?
• At which stage(s) in the production of sprouted seeds does contamination of the product with pathogenic microorganisms (e.g., Escherichia coli 0157, Salmonella, Bacillus cereus) occur?

• What methods are available to determine or predict that there is an increased risk that a production batch of sprouted seeds contains pathogenic microorganisms that can be used prior to harvesting the sprouts? For example:

• Are there any indicator organisms, quality attributes, or other product characteristics (e.g., scarification of seeds, geographical source of seeds) that can be used by industry or regulatory inspectors to predict an increased risk of contamination?
• What is the potential for irradiation to reduce incidence and prevalence of pathogenic microorganisms on sprouted seeds or the seeds from which they are produced?

• What is the effect of modified atmosphere packaging (MAP) and vacuum packaging (VAC) on growth and survival of pathogenic (including spore-formers) and non-pathogenic bacteria in sprouted seeds? In products irradiated under the conditions defined above? In products treated by different means (e.g., sanitizing agents)?
• Are there other potential alternative intervention technologies for reducing pathogens present on sprouted seeds or the seeds from which they are produced? What are the effects of these interventions on germination? What are the relevant treatment parameters that have to be considered in developing criteria for their effective use?

JUICE:

The primary research needs involve the identification and assessment of the efficacy of alternative treatments or strategies for the reduction or elimination of pathogenic microorganisms in fruit and vegetable juices, and the use of this information to provide guidance both to our inspectors and the industry.

• Are there simple, non-thermal treatments for the reduction or elimination pathogens in various fruit and vegetable juices that are amenable for use by small processors?
• Are there various nonpathogenic microorganisms (e.g., Lactobacillus fermentans, Enterobacter aerogenes) that are sufficiently similar to pathogens such as Salmonella Hartford or E. coli 0157:H7 with respect to attachment to fruit and equipment, heat resistance, acid tolerance, ability to survive at treatments with sanitizing agents, etc that they can be used to validate the effectiveness of antimicrobial treatments and processes for the reduction of pathogens during juice production in commercial operations (i.e., reliably serve as surrogate organisms)? What are the gaps, if any, in the safety data for the most promising candidate surrogate organisms?
• What levels of reduction, in specific pathogens, can be achieved through the use of various antimicrobial treatments (e.g., pulsed light, ultraviolet light, pulsed electric fields, ozone, etc.)? What are the relevant treatment parameters (e.g., temperature, pH, radiation dose, field strength, concentrations, etc) needed to achieve specified reductions (e.g., 1 log, 3 logs)?
• What is the effect of various factors, especially temperature, on the survival and growth of specific pathogens (or appropriate surrogate organisms) in finished juice that has not been pasteurized or otherwise been treated to reduce pathogens? In finished juice that has been treated to reduce pathogens by the various alternative antimicrobial treatments listed above?

GRAINS AND RELATED PRODUCTS

The primary foci are (1) to acquire data needed to assess the public health risks posed by dietary exposures to specific mycotoxins, and (2) to develop for field investigators and laboratories improved means for differentiating foods that have increased risks of significant contamination by mycotoxins.

• How can we improve our estimate(s) of human dose-response relations for the Fusarium-derived mycotoxins to which humans can be exposed through consumption of grain and related products?

• Are there techniques, data, or animal models available to assess/estimate human dose-response relations for specific mycotoxins to which humans can be exposed through consumption of grain, related food products, and other foods?

• How can monitoring of foods for mycotoxin contamination be improved?

FOODS OF ANIMAL ORIGIN

DAIRY PRODUCTS:

The following research areas reflect the need to evaluate potential food safety concerns that have emerged for which there are insufficient data to assess the risks to public health such that scientifically sound guidance can be provided to the inspection force and the industry.

• Is a 60-day aging period for raw milk cheese adequate to ensure eliminate of non-spore forming pathogenic bacteria, such as E. coli O157:H7 or L. monocytogenes? If not, what alternative production procedures will minimize the risk of foodborne illness resulting from such pathogens?
• Should FDA be concerned about the presence of Mycobacterium paratuberculosis in pasteurized milk or products made from pasteurized milk?

EGGS:

The regulatory focus with egg and egg products involves the need (1) to provide guidance to the industry and our inspectors concerning the effectiveness of strategies for preventing or eliminating contamination of shell eggs and egg products and (2) to reduce the costs and increase the effectiveness of laboratory techniques in support of field investigations and product tracebacks.

• Can analytical methods and sampling procedures be developed to more reliably and cost-effectively detect Salmonella Enteritidis when present in eggs at both a low frequency and low level of contamination (e.g., 1 infected egg containing <50 salmonellae in a pooled sample of 500 eggs)?
• Are there effective means for reducing the levels of S. Enteritidis and other salmonellae in intact shell eggs while maintaining the "raw character" of the food? What are the pertinent parameters that influence the effectiveness of the process both in relation to food safety and food quality?
• How do environmental parameters (e.g., temperature, pH, salt concentration, water activity, oxygen tension) affect the inactivation kinetics of foodborne pathogens by different intervention technologies proposed for use with liquid eggs and egg products? How do these parameters differ among different strains of pathogens such as S. Enteritidis and other salmonellae that can be internalized as a result of transovarian transmission?
• What levels of pathogen reduction in shell eggs can be achieved via non-thermal treatments? What are the relevant treatment parameters for achieving significant reductions in the levels of S. enteritidis without adversely affecting sensory acceptability and nutritional quality of eggs?

SEAFOOD

Molluscan Shellfish:

Priority research needs associated with raw molluscan shellfish are related to the recent emergence of Vibrio parahaemolyticus as an cause of disease in consumers of raw molluscan shellfish. The research areas reflect a need to provide state and federal officials with means (1) to assess the risks associated with this microorganism and (2) to make scientifically sound decisions related to the management of risks associated with consumption of raw molluscan shellfish.

• What levels of tdh⁺ V. parahaemolyticus strains (V. parahaemolyticus strains that have the gene that encodes for thermolabile direct hemolysin) are found in raw molluscan shellfish at retail?
• What is the infectious dose (or probability of infection) for tdh⁺ V. parahaemolyticus? Are there specific serotypes/strains that have increased infectivity/virulence? How does the infectivity of tdh⁺ V. parahaemolyticus vary among various subpopulations, particularly those at increased risk?
• What means are available that federal and state regulators can used to determine when there is an unacceptable risk of tdh⁺ V. parahaemolyticus outbreaks from the consumption of raw molluscan shellfish?

• What handling techniques or processing treatments (e.g., heat shock, irradiation, other) are effective for reducing and/or eliminating V. parahaemolyticus and V. vulnificus from raw molluscan shellfish?
• Are there microbiological, chemical, or physical attributes (e.g., coliphage levels, water temperature or salinity) that can be used to predict an increased risk of contamination of molluscan shellfish with viruses of public health concern (e.g., Norwalk viruses, hepatitis type A).

Fin Fish and Crustaceans:

The focus of research related to fin fish is related to the need to provide guidance to the seafood industry, our inspection forces, and the developers of food safety policies on (1) means for preventing scombrotoxin intoxications, (2) the food safety implications of employing different forms of product packaging, (3) approaches for measuring and reducing the risks associated with the consumption of raw fin fish and crustaceans, and (4) the likelihood that aquiculture products are a source of pathogenic bacteria that are resistant to multiple antimicrobials.

Scombrotoxin/Biogenic Amines/Decomposition

• What harvesting/processing practices increase the potential for scombrotoxin production and what corrective measures will prevent such occurrences?
• What indicators (e.g., sensory, chemical) can be used effectively by inspection personnel to differentiate seafood with increased risk of contamination by scombrotoxin?
• Are time/temperature integrators currently available sufficiently accurate to identify fish that have been held under storage conditions conducive to the formation of scombrotoxin?
• What correlation, if any, exists between sensory attributes used to identify decomposition and the formation of cadaverine and putrescine? Histamine? Are putrescine and cadaverine themselves toxic at the levels in which they are present in decomposed seafood or are they potentiators of toxicity induced by another agent?
• Why does decomposition in non-scombroid species of fish (e.g., salmon) and other seafood sometimes cause illness with symptoms similar to scombroid poisoning? Do microbial toxins other than biogenic amines play a role in outbreaks of scombrotoxicity?
• Can decomposition be reliably and objectively determined by analytical methods? What part of the fish is the best indicator that decomposition has occurred?
• Does carbon monoxide treatment of scombroid fish pose a hazard to consumers due to CO-treatment masking of the visual or other sensory signs of decomposition currently used as indicators of possible scombrotoxin formation? Is there a reliable field method to rapidly determine if fish, especially tuna, have been treated with CO?

Packaging Technologies

• Is spoilage before toxin formation a suitable control mechanism for MAP and VAC products?
• Do MAP and VAC of seafood increase the growth of pathogenic bacteria and/or toxin formation (esp. C. botulinum) before organoleptic signs of spoilage are apparent?
• What packing conditions (e.g., oxygen tension, pH, water activity) establish conditions that favor growth and toxin production by "anaerobic" pathogens?

Handling and Preparation of Raw Fin Fish and Crustaceans

• What is the incidence of parasitic illness in the United States that is attributable to the consumption of fin fish?

• What environmental and production parameters must be maintained to ensure that outgrowth of, and toxin production by, Staphylococcus aureus does not occur during manufacturing of commercial breaded/battered seafood products (e.g., frozen fin fish, shrimp)?
• Do commercially defined time-temperature processing schedules consistently reduce and/or eliminate bacterial pathogens of concern from seafood products? For example, what cumulative time-temperature exposures are needed to ensure control of bacterial hazards in commercial cooked blue crab meat?
• How can levels of Listeria monocytogenes on cold smoked salmon be reduced?
• Are the cooking practices used in the home and food service establishments sufficient to inactivate pathogenic bacteria, viruses, or parasites that may be present in fin fish?

Antimicrobial Resistance

• What approved and unapproved antimicrobials (antibiotics and antibiotic-like compounds) and their residues are present in imported and domestic aquiculture seafood? Do aquiculture products represent a significant risk in relation to the emergence of antimicrobial resistant foodborne pathogens in human populations?

SPECIAL NUTRITIONALS

The focus of the research is the acquisition of data needed to better assess the microbial food safety risks that may be associated with (1) the sporadic, low levels of certain microorganisms found in infant formulas and other special nutritionals and (2) the use of viable microorganisms as dietary supplements.

• What is the microbial food safety significance of non-spore forming gram-negative bacteria (e.g., Enterobacter sakazaki) that are found sporadically at low levels in powdered infant formulas? Are there more effective and cost-effective sampling methods for these microorganisms in powdered infant formulas?
• What is the microbial food safety significance of low levels of B. cereus and other spore forming microorganisms in infant formulas? What is the potential for heat stable enterotoxins, such as that produced by B. cereus, to be carried into finished powdered infant formulas if present in the ingredients?
• What data are needed to assess the risk posed by potential contaminants (TSE infective agents and microbial pathogens) in glandular products of animal origin and meat byproducts that are used as dietary supplements?
• What is the risk that microorganisms used as probiotic (e.g., Lactobacillus spp., blue-green algae) dietary supplements could cause infections in individuals that have severely compromised immune systems.

ESTIMATING THE RISK AND IMPACT
OF FOODBORNE DISEASE

• How can the findings from epidemiological investigations of outbreaks of foodborne illness be used to draw the most accurate estimates of both background risk and cost of foodborne disease and "attributable risks and costs," i.e., the risk and cost of disease from a particular microorganism as a result of eating a particular food?
• Are there alternative means for deducing dose-response relations for foodborne pathogens that can be used as an alternative to human volunteer feeding studies? Animals?
• What segments (e.g., neonates, the elderly) of the general population should be considered at increased risk for foodborne disease? How does this vary for different disease agents?
• Are there alternative approaches to the validation of new chemical and microbiological methods that would be more cost effective and timely than existing collaborative study protocols, while still maintaining an equivalent level of assurance? What are the relative costs of these alternative procedures?
• How can exposure data on foodborne pathogens that occur only sporadically and at low levels be acquired in a more effective and cost-efficient manner? How can the current ability of the industry to control the presence of foodborne pathogens in various foods be assessed? How can improvements in microbial food safety be assessed objectively and cost-effectively?
• Are there quantitative data available on the extent of transfer of pathogenic bacteria from the hands of food workers to food items? How is this rate of transfer influenced by food type, frequency of hand washing, and other parameters? Are there differences in transfer rates when a food worker is the source of contamination versus when they are the means of cross contamination between raw and ready-to-eat foods?

EMERGENCE OF NEW MICROBIAL CONCERNS

The focus of this research area is the need to enhance the ability of FDA's food safety program to respond rapidly to the emergence or re-emergence of new microbial threats through the acquisition of knowledge in microbial genetics and physiology needed (1) to rapidly characterize new and/or emerging foodborne pathogens, (2) to facilitate methods development, and (3) to identify predictors of new microbial threats.

• Are there common virulence characteristics that are likely to be transferred among foodborne microorganisms that can be used as molecular targets to develop detection reagents and methods for emerging pathogens.
• Are there means for estimating the dose-response relations for a newly emerged foodborne pathogen based on the integration of its virulence characteristics?
• What impact do food production, processing, and marketing operations have on the frequency of genetic and physiological changes that would foster the emergence or re-emergence of pathogens with increased potential to cause disease?

DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration
Washington, DC 20204
November 19, 1998

Dr. Eileen Kennedy
Deputy Under Secretary
Research, Education and Economics
United States Department of Agriculture
Whitten Building, Room 217-W
1400 Independence Avenue, SW
Washington, DC 20250

Dear Dr. Kennedy:

As part of the Food Safety Initiative research planning for the 1999 fiscal year and in anticipation of the initiation of the Joint Institute for Food Safety Research, our Center Director, Joseph Levitt, asked the FSI Research/Risk Assessment staff to develop and communicate to you the enclosed list of research needs. They represent priority research needs associated with the Food and Drug Administration's mission to develop and implement scientifically sound food safety policies for the foods for which our agency is responsible. In developing this list of priority research needs, we have focused primarily on pre-harvest research areas that the intramural and extramural programs of the USDA research agencies have unique facilities and expertise, or for which we know that there is USDA research already underway. Without question, USDA research programs can achieve significant gains in these areas much more quickly and efficiently than FDA attempting to duplicate USDA's research strengths. Some limited requests in FDA-specialized areas, where our current/expected research-related resources are just not sufficient, are also included.

We will be happy to provide you a detailed briefing on any or all of the research areas identified. Likewise, we are always pleased to meet with representatives of REE and its member agencies to discuss and coordinate our research needs, goals, and plans.

Thank you in advance for your consideration.

Sincerely,

Robert L. Buchanan, Ph.D.

Lead Scientist for Food Safety Initiative

Enclosure cc:
Dr. Catherine Woteki, Under Secretary for Food Safety, USDA
Dr. Kaye Wachsmuth, Deputy Administrator for Public Health and Science,
Food Safety and Inspection Service, USDA
Dr. Colien Hefernan, Acting Administrator, CSREES, USDA
Dr. William Raub, Assistant Secretary for Health Policy, DHHS
Dr. James O'Hara, Under Secretary for Public Health and Science, DHHS
Dr. Michael Friedman, Lead Deputy Commissioner, FDA
Mr. Joseph Levitt, Director, CFSAN, FDA
Ms. Janice Oliver, Deputy Director for Systems and Support, CFSAN, FDA
Mr. Keith Pitts, Special Assistant to Secretary Glickman, USDA
Mr. Eric Olsen, Special Assistant to Deputy Secretary Rominger, USDA

Priority FDA Research Needs for the USDA Research Agencies

A. Microbiological Safety of Produce

1. Determine the incidence and prevalence of foodborne pathogens on various fruits and vegetables, and identify factors (e.g.,regional/ seasonal differences, climate, damage during harvest) that contribute to the level and persistence of the pathogens.

2. Assess the potential for foodborne pathogens to grow or survive in or on fruits and vegetables, including evaluation of varietal differences and the effects of storage/transport temperatures (e.g., identify temperatures that minimize or inhibit pathogen growth).

3. Develop guidelines for the proper storage, treatment, and reuse of containers used to harvest fresh produce for raw consumption. Containers are often stored outside where contamination by birds and pests is possible. Containers made of different materials should be considered (i.e, plastic, wood, cloth).

4. Assess the potential for using produce quality attributes as indicators of increased risk of contamination by foodborne pathogens.

5. Identify factors that can lead to the internalization of foodborne pathogens by various fruits and vegetables (e.g. infiltration, contamination at time of flowering, injury).

6. Determine the factors influencing the microbiological quality of agricultural and greenhouse water and develop simple field methods for its assessment.

7. Develop inexpensive, practical means for improving the microbiological quality of agricultural and greenhouse water.

8. Assess the effect of locating animal production facilities near or adjacent to produce growing areas on the incidence and prevalence of foodborne pathogens on or in various classes of fruits and vegetables. Research should also include the assessment of mechanisms of spread by pathogens (e.g., wind-drift, runoff).

9. Develop quantitative models for the survival and inactivation of foodborne pathogens in manure and the development of "user friendly" guidelines for the safe use of manure.

10. Develop means for accelerating the elimination of pathogens via the composting of manure, and develop a simple means for farmers or manure suppliers to monitor the effectiveness of the process.

11. Assess the role insects, birds, and feral animals may have as vectors for the contamination of fruits and vegetables by foodborne pathogens. Assess the impact of practices, including the planned introduction of domestic animals into produce growing areas, on the incidence of pathogens on or in produce (e.g., turning cattle into a field to glean residue after harvest; geese in an orchard to control weeds or snails).

12. Develop intervention technologies for use at harvest that reduce the risk of foodborne pathogens on field-packed commodities.

13. Develop intervention technologies that can be used in packing houses or processing facilities to reduce or eliminate pathogens in or on fresh or fresh-cut produce.

14. Identify non-human reservoirs for Cyclospora and the sources, vectors, and agricultural practices that contribute to its presence on produce.

B. Sprouted Seeds

1. Develop intervention technologies that can be used at seed mills to reduce the incidence and prevalence of foodborne pathogens on seeds destined for the production of sprouts (also relate to water quality concerns. . .i.e., see A.6).

2. Assess the effect of scarification on the potential for seeds intended for the production of sprouts to harbor foodborne pathogens.

3. Assess the potential for using seed quality attributes, including varietal differences, as indicators of increased risk of contamination by foodborne pathogens.

4. Develop means for retarding the growth of foodborne pathogenic bacteria during the germination and outgrowth of sprouted seeds.

5. Develop intervention technologies that can be used just prior to germination to reduce the prevalence of foodborne pathogens on seeds to be used for sprouting.

C. Dairy Products

1. Determine the incidence and levels of Listeria monocytogenes, Salmonella spp., Mycobacterium paratuberculosis, Escherichia coli 0157:H7, and Cryptosporidium parvum in farm bulk tank raw milk, and identify factors (e.g., seasonal and regional differences, husbandry practices, farm ecology of the pathogen) that contribute to contamination by these pathogens.

D. Agricultural Antimicrobial Use

1. Develop a database of prudent practices for use of antibiotics in food animals (and aquaculture; see F4) that can be used to conduct educational research to determine the most effective means to educate food animal producers on proper use of antibiotics.

2. Quantify antibiotic resistance development on swine farms using specific drugs as part of herd health programs, including following the animals through slaughter to determine the incidence and prevalence of antibiotic resistance in both the general microflora and pertinent foodborne pathogens on meat entering commerce.

3. Conduct an Antimicrobial Prescription Survey to determine prescribing practices of food animal practitioners, production practices among the cattle and swine industries, and the relationship between antimicrobial use and resistance on the farm.

4. Conduct feasibility study for the development, implementation, and maintenance of an international database on antimicrobial resistance.

5. Determine the basal antibiotic resistance patterns in dairy cattle, and assess the persistence of those patterns after clinical outbreaks, including the effect of therapeutic drug treatment on the pattern of antibiotic resistance.

E. Meats, Game Meats, or Poultry

1. Assess the extent of implementation of animal feed rules for the prevention of the spread of Bovine Spongiform Encephalopathy (BSE) in the U.S.

2. Develop and employ molecular methodologies for the detection of Clostridium perfringens in poultry production facilities to facilitate identification of on-farm sources of contamination. Research should emphasize antibiotic resistance of this pathogen.

3. Develop methods to detect game meats (e.g., wild deer and elk) contaminated with transmissible spongiform encephalopathies (TSEs) agents. Determine the incidence of game animals with TSEs that are consumed by human populations in the United States. Both data sets will aid in calculating the likelihood for producing TSEs in humans for risk assessment.

F. Aquaculture

1. Identify the types and sources of bacteria typically found in closed, recirculating systems used in aquaculture, with a particular emphasis on the incidence and prevalence of potential foodborne pathogens (e.g., most notably Listeria monocytogenes and Salmonella spp., especially the latter contaminating aquacultured shrimp). Develop appropriate controls to avoid pathogen contamination of aquacultured products by these pathogens.

2. Develop models relating environmental conditions to the potential for unacceptable levels of Vibrio parahaemolyticus and Vibrio vulnificus and other foodborne pathogens in aquaculture fish, crustaceans, and shellfish from aquatic and estuarine environments.

3. Determine the sources of foodborne pathogens in freshwater aquaculture products, and identify environmental factors and production practices that influence the incidence and prevalence of foodborne pathogens both in the growing environment and the harvested animals.

4. Develop methods for detecting aquaculture drugs (antibiotics)/chemicals, or appropriate metabolites thereof, to provide appropriate regulatory controls.

G. Egg and Egg Products

1. Develop prophylactics, such as vaccines or competitive exclusion, to eliminate or minimize Salmonella enteritidis from laying flocks.

2. Determine the dynamics of Salmonella enteritidis in a naturally infected flock (e.g., spread through the house; environmental positivity versus egg positivity; recurrence of infection over the lifetime of a flock which has included a molt).

3. Develop the means for differentiating layer flocks where Salmonella enteritidis remains in the intestinal tract versus those flocks where it resides in the reproductive tract or in both the reproductive and intestinal tracts, resulting in production of eggs internally contaminated with the pathogen (i.e.,virulence of pathogen and genetics of breeds).

4. For Salmonella serovars other than Salmonella enteritidis that infect reproductive tracts (e.g., S. heidelberg, S. typhimurium),determine virulence factors, genetics of breed, production, feed or environmental selection pressures that may result in increased transovarian infectivity, emphasizing serovars resistant to multiple antibiotics.

5. Develop alternative, non-thermal treatments for the elimination of pathogens from shell eggs and liquid egg products.

H. Grains and Oilseeds

1. Develop grain and oilseed varieties that are resistant to the formation of mycotoxins, particularly fumonisins, vomitoxin, and aflatoxin.

2. Determine the factors that influence the frequency and extent of fumonisin contamination in both sweet and field corn. Determine the consumption patterns for sweet corn in the human diet. Develop data sets based on fumonisin contamination and corn consumption patterns that will support the development of risk assessments for fumonisin.

3. Review data on cellular and molecular responses observed in animals after exposure to fumonisin and conduct similar experiments using appropriate human cells (e.g., macrophages or endothelial cells), to indicate whether humans are more similar to horses or rodents.

4. Develop and evaluate biomarkers for human exposure to fumonisins and resulting biological effects.

5. Determine the factors that influence the frequency and extent of contamination of grains and oilseeds by aflatoxins, fumonisins, deoxynivalenol, acetyldeoxynivalenol, nivalenol, ochratoxin, cyclopiazonic acid and zearalenone to obtain sufficient exposure data for use in conducting risk assessments.

6. Evaluate the potential immunotoxic, neurotoxic, and reproductive effects of trichothecene mycotoxins.

7. Develop rapid, sensitive, cost-effective analytical methods for the determination of mycotoxins for use in monitoring and control programs.

8. Assess the potential for Bacillus cereus growth and toxin production on cooked grain-based foods, including during storage of the cooked food.

I. Juices

1. Develop intervention technologies, suitable for use by small juice manufacturers, that eliminate foodborne pathogens without thermally processing the juice.

J. Consumer research in support of food safety information policies.

1. Identify population and subgroup trends for selected food safety knowledge, attitudes and practices (e.g., risky food consumption behaviors, in-home food preparation and handling behaviors) that may be significant behavioral risk factors for foodborne illness and may be targeted as such be food safety education and labeling policies.

2. Evaluate the consumer impacts of ongoing food safety information policies such as USDA care labels on meat and poultry products, warning statements on unpasteurized juices, safe handling instruction on shell eggs, the Fight Back education campaign and other labeling and education initiatives.

3. Conduct validation studies to develop and standardize measures of consumer food handling practices, perceptions of food safety risks, and consumer confidence.

4. Evaluate the motivational bases and barriers to change of consumer compliance with food safety recommendations.

5. Conduct special surveys of high-risk groups such as pregnant women who are too rare in the population to be adequately reached with general population surveys to assess knowledge, attitudes and practices related to their special needs, such as the risk of L. monocytogenes for neonatal health.

K. General

1. Assess the role of transportation practices as a source of cross contamination of commodities, and develop improved sanitation practices, based on identification of sources of contamination during transportation of food and food animals from production locations (i.e., farms) to initial processing facilities (e.g., slaughter house, packing house, grain milling).

2. Determine the incidence and prevalence of abnormal levels of biogenic amines and their reaction products in foods associated with human and animal adverse reactions and identify factors that contribute to these levels.

Appendix B

Descriptions of FDA/FSI-funded Grants

1. Risk Assessment Grant #: FD-R-001621: Establish a Correlative Dose-Response Model For Human Cryptosporidiosis Principal Investigator: Saul Tzipori, School of Veterinary Medicine, Tufts University

2. Risk Assessment Grant #: FD-U-001622-01: Development of a Risk Assessment Dose-Response Model for Food borne Listeria monocytogenes Principal Investigator: Mary Alice Smith, University of Georgia

3. Risk Assessment Grant #: FD-R-001625-01: Dose-Response to Vibrio Species Principal Investigator: J. Glenn Morris, Jr., University of Maryland

4. Research Grant #FD-U-001626-01: Disinfection of Alfalfa Seeds and Sprouts Principal Investigator: Larry R. Beuchat, Ph.D., University of Georgia

5. Research Grant #FD-U-001631-01: Inactivation of Pathogens on Produce by GRAS Chemicals Principal Investigator: Michael P. Doyle, Ph.D.,University of Georgia

6. Research Grant #FD-U-001638-01:A Non-Thermal Method to Enhance Safety of Fresh Produce Principal Investigator: Yen-Con Hung, Ph.D., University of Georgia, GA

7. Research Grant #FD-U-001627-01: Natural Variation in Escherichia coli O157:H7 Principle Investigator: Frederick Blattner, University of Wisconsin

8. Research Grant #FD-U-001629-01: Consumer Handling and Washing of Fresh Fruits and Vegetables Principal Investigator: Linda Joan Harris, Ph.D., UCLA