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Campylobacter hepaticus  causes Spotty Liver Disease (SLD) in chickens.  C. hepaticus  is fastidious and slow-growing, presenting difficulties when growing this bacterium for the preparation of bacterin vaccines and experimental disease challenge trials. This study applied genomic analysis and in vitro experiments to develop an enhanced  C. hepaticus  liquid culture method. In silico analysis of the anabolic pathways encoded by  C. hepaticus  revealed that the bacterium is unable to biosynthesise  l- cysteine,  l- lysine and  l- arginine. It was found that  l -cysteine added to Brucella broth, significantly enhanced the growth of  C. hepaticus , but  l- lysine or  l- arginine addition did not enhance growth. Brucella broth supplemented with  l- cysteine (0.4 mM),  l- glutamine (4 mM), and sodium pyruvate (10 mM) gave high-density growth of  C. hepaticus  and resulted in an almost tenfold increase in culture density compared to the growth in Brucella broth alone (log10 = 9.3 vs 8.4 CFU/mL). The type of culture flask used also significantly affected  C. hepaticus  culture density. An SLD challenge trial demonstrated that  C. hepaticus  grown in the enhanced culture conditions retained full virulence. The enhanced liquid culture method developed in this study enables the efficient production of bacterial biomass and therefore facilitates further studies of SLD biology and vaccine development.

Enhancement of Campylobacter hepaticus culturing to facilitate downstream applications