Saeed A. Khan, Ph.D. Division of Microbiology National Center for Toxicological Research US Food and Drug Administratio

1. National Center for Toxicological ResearchScientific Advisory Board Subcommittee Program Review of the Division of Microbiology “Response to the Subcommittee Report” Saeed A. Khan, Ph.D. Division of Microbiology National Center for Toxicological Research US Food and Drug Administration Jefferson, Arkansas

2. National Center for Toxicological Research Advisory Board Site Visit Review Team Consumer Representative Dr. Robert E. AndersonFDA Liaison MembersDr. Peter H. Cooney (CBER) Dr. Roger A. Jones (CVM) Dr. Arthur J. Miller (CFSAN) Dr. David G. White (CVM) Science Advisory BoardMember Dr. Catherine W. Donnelly, ChairConsultants To Advisory Board Dr. Raul J. Cano Dr. Thomas W. Federle Dr. Mike G. Johnson Dr. Susan F. Kotarski

3. Comments on the General Recommendations • Strategic Planning • A New Philosophical Paradigm • Leadership • Building Capacity • Public Relations/Communications

4. Comments on the Research Focal Areas • Foodborne Pathogens, Food Safety and Methods Development • Determination of the Role of Intestinal Microflora In Activation or Detoxification of Xenobiotics • Environmental Biotechnology

5. Comments on the Research Focal Areas (Cont.) • Use of Microorganisms as Models to Predict the Metabolic Pathway by which Drugs are Metabolized in Animals • Microbiologist Surveillance/Diagnostic Support of Research

6. Overview of the Division of Microbiology Saeed A. Khan, Ph.D. Division of Microbiology National Center for Toxicological Research US Food and Drug Administration Jefferson, Arkansas

7. Division of Microbiology Division of Chemistry Department of Health and Human Services (DHHS) Food and Drug Administration (FDA) Office of Operations National Center for Toxicological Research Office of the Director Office of the Deputy Director for Research Division of Genetic and Reproductive Toxicology Genetic Toxicology Lab Reproductive Toxicology Lab Division of Biometry and Risk Assessment Division of Biochemical Toxicology Division of Molecular Epidemiology Division of Neurotoxicology

8. Mission The Division of Microbiology at the NCTR serves a multipurpose function with specialized expertise to perform fundamental and applied research in microbiology in areas of FDA’s responsibility in toxicology. The Division of Microbiology also responds to microbial surveillance and diagnostic needs for research projects within the NCTR and FDA.

9. Virology Serology Division of Microbiology Director Surveillance/Diagnostic Program Research Program Environmental Biotechnology Media Preparation Food Safety Microbial Models in Toxicology Intestinal Microflora Projects Bacteriology Parasitology Mycology Center Surveillance and Research Support FDA/Division of Microbiology Research Collaboration Breeding Colony Quarantine Facility Primate colony NTP Animal Studies NON-NTP Animal Studies Animal Husbandry Contractor Diet Preparation Contractor Environmental Health and Program Assurance Microbiology Division Biochemical Toxicology Division Chemistry Division Genetic & Reproductive Toxicology Division Molecular Epidemiology Division Neurotoxicology Division

10. Division of Microbiology Organizational Chart Administration Cerniglia, Carl E., Ph.D., Director, Division of Microbiology Fleischer, Patricia Malone, Sandra L. Research Program Principal InvestigatorsIntern Program Erickson, Bruce D., Ph.D. Bunch, Sherita Khan, Ashraf A., Ph.D. Wynne, Rebecca Khan, Saeed A., Ph.D. Nawaz, Mohamed, Ph.D. Technical Support Pothuluri, Jairaj V., Ph.D. Moody, Joanna D. Rafii, Fatemeh, Ph.D. Williams, Anna Sutherland, John B., Ph.D. Wang, Rong-Fu, Ph.D. ORISE Cha, Chang-Jun, Ph.D. Kim, Seong-Jae, Ph.D. Kim, Yong-Hak, Ph.D. Nayak, Rajesh, Ph.D. Surveillance/Diagnostic Program Campbell, Warren, Program Leader Camp, Marvin W. Colvert, Ronald M. Holland, Michael A. Paine, Donald D. Robertson, Latriana H. Steele, Roger S. Summage-West, Christine Wagner, R. Doug, Ph.D. (PI) Williams-Sims, Lillie M.

11. Anaerobic Bacteria Culture and Detection • Bacteria Isolation and Identification • Cellular Fatty Acids Analysis (MIDI) • Diagnostic Surveillance of Environmental Samples • Endo and Ecto-parasite Identification • Fungal Isolation and Identification • Media Preparation Facility • Rapid Molecular Methods in Detection of Microorganisms • Virus Detection • PCR • DNA Sequencing • ELISA • Gene Cloning • Gene Isolation and Manipulation • Hybridoma Technology • Monoclonal and PolyclonalAntibodies • Site-directed Mutagenesis Division of Microbiology Staff Skills and Facilities • Semicontinuous Culture System • Fermentation Facility • Microbial Bioassays • Tuberculocidal Assays of Chemical Germicides • Protein Purification Techniques • Biodegradation Methods • Isolation and Identification of Chemical Intermediates • Gas liquid Chromatography • High Performance Liquid Chromatography

12. Division of Microbiology FDA Projects   CFSAN    ORA  3  NCTR Microbiology 6    CVM   CDRH 10   1   CBER  CDER   

13. Microbiology Division ResearchProjects with the Food Safety Initiative • In Vitro Model and Molecular Analysis of Competitive Exclusion Products • Molecular Screening Methods for the Determination of Vancomycin Resistance in Selective Competitive Exclusion Products. • Studies on the Fluoroquinolone Resistance in Campylobacter sp. isolated from Poultry

14. Microbiology Division ResearchProjects with the Food Safety Initiative • Studies on Mechanism of Fluoroquinolones Resistant Salmonella spp. Isolated from Poultry. • Screening Animal Feeds Using Molecular Techniques for Proteins Derived from Mammalian Tissue to Detect Possible Bovine Spongiform Encephalopathy (BSE) • Molecular Methods to Identify and Quantitate Human Foodborne Pathogens in the Animal Production Environment. • Biodegradation of Veterinary Drug Residues.

15. Recent Accomplishments • Developed • Molecular techniques for detecting (a) human intestinal microflora (b) 13 different strains of food-borne pathogens (c) animal pathogens and environmental isolates • Guidance document entitled “Assessing the effects of antimicrobial residues in food on the human intestinal microflora” • Alternative microbial methods for studying metabolism of drugs

16. Recent Accomplishments (Cont.) • Developed • Bioluminescent assay to determine the tuberculocidal activity of disinfectants • Developed multiplex PCR methods for the determination of vancomycin resistance genes • Developed a cell-based model of colonization resistance against Salmonella invasion • Developed a multiplex PCR method for the detection of Salmonella typhimurium DT104.

17. Recent Accomplishments (Cont.) • Characterized • Microbial cytochrome P-450s and glutathione transferase • Aerolysin toxins genes • Erythromycin resistance genes (erm) in Staphylococcus sp. isolated from chickens • Azoreductase and nitroreductase enzymes in bacteria isolated from human intestinal tract • Fluoroquinolone resistant Campylobacter spp. from poultry samples.

18. Recent Accomplishments (Cont.) • Characterized competitive exclusion products. • Completed method validation trial for the detection of BSE • Determined the survival of Shigella on foods • Discovered an agent in oyster homogenates that is lethal to V. cholerae and V. vulnificus • Discovered new strains of bacteria and fungi that degrade environmental pollutants • Elucidated novel biodegradation pathways for xenobiotics

19. FY 2001 Plans PI: Bruce Erickson, Ph.D. Initiate a new chemostat experiment for testing veterinary fluoroquinolones. PI: Ashraf Khan, Ph.D. Isolation and molecular characterization of fluoroquinolone resistant Salmonella spp. and E. coli from chicken and turkey litters.

20. FY 2001 Plans (Cont.) PI: Saeed Khan, Ph.D.Study the plasmid profile of vancomycin- resistant bacteria isolated from Competitive Exclusion Products. Genetic fingerprinting and strain typing of vancomycin-resistant organisms isolated from the CE product will be carried out by pulse- field gel electrophoresis and molecular probing.

21. FY 2001 Plans (Cont.) PI: Mohamed Nawaz, Ph.D. Correlate the environmental data with the occurrence of fluoroquinolone resistance to campylobacters. Characterize all fluoroquinolone resistance campylobacters at the molecular level (PCR-RFLP, PFGE). PI: Jairaj Pothuluri, Ph.D. Determine the biodegradation rates and metabolic fate of the antibiotics used in aquaculture.

22. FY 2001 Plans (Cont.) PI: Fatemeh Rafii, Ph.D. Detect specific bacteria from the human intestinal tract involved in the conversion of phytoestrogens (daidzein and genistein) to estrogenic and non estrogenic end products. Evaluate the effect of fluoroquinolones on resistance development in anaerobic bacteria from the human intestinal tract, mechanism of resistance development, impact on metabolic activities and the dissemination of resistance to bacterial pathogens.

23. FY 2001 Plans (Cont.) PI: John Sutherland, Ph.D. Identification of the metabolites produced from norfloxacin and sarafloxacin by fungi grown on poultry litter. Isolation of new strains of fungi from litter in poultry houses and screening them for the biotransformation of veterinary fluoroquinolones. PI: Robert Wagner, Ph.D. Complete in vitro assay of competitive exclusion products.

24. Vision Strive for scientific excellence and strengthen the relevance of the research in the Division of Microbiology with the mission of the Food and Drug Administration. Maintain a world-class research program to solve current issues that face the FDA in the next millennium, so the Agency can make sound science-based regulatory decisions on microbiology.