2008 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to perform genome analysis of Leptospira species to identify genetic differences and variation in gene expression under different experimental conditions. A variety of approaches will be used, including genome sequencing, bioinformatics, development of microarrays, and analysis of proteomes. 1b.Approach (from AD-416) Each laboratory independently determined the genome sequence of strains of Leptospira borgpetersenii serovar hardjo. In addition, the laboratory at Monash University has revised the annotation of two Leptospira interrogans genomes. Comparative sequence analysis using bioinformatics will be used to identify alterations in genes and attempts will be made to correlate these genetic differences with altered characteristics of the bacteria. Genome sequence data will be used to design and produce microarrays that will be used to identify changes in gene expression under different environmental conditions. Protein expression patterns will be determined using proteomic analysis techniques and immunological analysis. 3.Progress Report In FY2008, studies on microarray analysis of the Leptospira interrogans response to iron limitation were conducted by collaborators. A poster session describing these results was presented at an international meeting on bacterial genomics. Genomic analysis of Leptospira biflexa was completed leading to the discovery of three sets of genes: those unique to non-pathogenic saprophytes, those unique to pathogenic Leptospira, and genes shared among both saprophytic and pathogenic strains. This work was submitted for consideration and published. Comparative sequence analysis of spirochetes is continuing; genomic analysis of the spirochete Brachyspira hyodysenteriae was conducted with the discovery of variation among a major surface protein family. A poster session was presented at the annual meeting of the Australian Society for Microbiology. Progress under this agreement is monitored by periodic conference calls and e-mails. Conference calls are organized at varying periods, often at least once a month, sometimes weekly. Data generation can be monitored by secure Internet access to a server hosted by the cooperator. This accomplishment aligns with the “countermeasures to prevent and control zoonotic diseases” action plan, of NP 103, component 3, problem statement 3B.