2007 Annual Report 1a.Objectives (from AD-416) The objectives of this project are to work with the Department of Fisheries and Allied Aquacultures of Auburn University to conduct research to characterize bacterial genomic sequences from fish pathogens for prediction of genes that may encode vaccine candidates. Further, the work includes epidemiological and diagnostic research. The work includes epidemiological research related to major fish health problems of catfish for vaccine development. Further, the work is to identify genes responsible for virulence of the protozoa Ich. 1b.Approach (from AD-416) Work closely with Department of Fisheries and Allied Aquacultures of Auburn University to conduct the pathogen genomic research, collecting, analyzing and reporting on data. By applying appropriate tools of bacterial protozoan genomic technologies to the sequences of candidate pathogens, it is possible to identify putative virulence factors for diagnostic and vaccine candidates. The major pathogenic bacteria to be sequenced include Edwardsiella ictaluri and Flavobacterium columnare and Ich. The utilization of genomic sequence in combination with the application of bioinformatics through genomics and proteomics can expedite the epidemiological diagnostic and vaccine discovery process by rapidly providing sets of potential candidates for further testing. Epidemiological and pathological research will also be conducted by application of diagnostic tests and field evaluation of fish pathogen problems. 3.Progress Report This report serves to document research conducted under a Specific Cooperative Agreement between Auburn University and the Aquatic Animal Health Research Unit. Additional details can be found in the in-house projects, 6420-32000-022-00D, "Aquatic Animal Diagnostics, Pathogenesis and Applied Epidemiology"; 6420-32000-019-00D, "Vaccinology and Immunology of Aquatic Animals"; and 6420-32000-020-00D, "Molecular Analysis of Virulence Determinants of Select Bacteria in Fish Diseases." Flavobacterium columnare molecular epidemiology Flavobacterium columnare is an important fish pathogens and information of genetic groups and fish species is lacking. Flavobacterium colunmare isolates were obtained from fish species present in the Mobile River and catfish farm ponds. The F. columnare isolates were fingerprinted using amplified fragment length polymorphism (AFLP) and intergenic spacer region (ISR) single-strand conformation polymorphism analysis. The majority (more than 85%) of the isolates recovered from catfish (including both blue and channel catfish species) were genomovar II. Flavobacterium columnare genomics Information on time differential gene expression in F. columnare is absence in the literature. A shot-gun genomic library of the Flavobacterium columnare ALG-530 virulent strain has been constructed and more than 3,000 clones have been sequenced. The expression profile of the genes tested varied when the strain was grown in vitro. Expression of the thiamine gene, glycosyltransferase gene, and gliding gene (gldB) was not detected. Gene expression level in the presence of catfish skin was lower than under other conditions. Discovery and characterization of lytic phages to Edwardsiella ictaluri Phage therapy is an alternative to vaccination for disease control. To date, we have identified several lytic phages to the catfish pathogen E. ictaluri and are characterizing those phages and their ability to control enteric septicemia of catfish in infected catfish. Parasite pathogen gene expression The ciliate protozoan Ichthyophthirius multifiliis (Ich) is an important parasite of catfish that causes 'white spot disease' leading to significant losses. A genomic resource for large-scale studies of this parasite has been lacking. To study gene expression involved in Ich pathogenesis and virulence, our goal was to generate expressed sequence tags (ESTs) for the development of a microarray platform for the analysis of global gene expression in this species. The results provide a basis for the development of miroarrays useful for gene expression studies or Ich development and virulence. Monitoring activities include meetings, phone calls, site visits, scientific mansucripts, and receipt of written progress reports, initiated by the Authorized Departmental Officer's Designated Representative (ADODR) or Cooperator, involving the exchange of information about the state of or progress of the agreement.