Arginine deiminase pathway is far more important than urease for acid resistance and intracellular survival in Laribacter hongkongensis: a possible result of arc gene cassette duplication.

Background Laribacter hongkongensis is a Gram-negative, urease-positive bacillus associated with invasive bacteremic infections in liver cirrhosis patients and fish-borne community-acquired gastroenteritis and traveler's diarrhea. Its mechanisms of adaptation to various environmental niches and host defense evasion are largely unknown. During the process of analyzing the L. hongkongensis genome, a complete urease cassette and two adjacent arc gene cassettes were found. We hypothesize that the urease cassette and/or the arc gene cassettes are important for L. hongkongensis to survive in acidic environment and macrophages. In this study, we tested this hypothesis by constructing single, double and triple non-polar deletion mutants of the urease and two arc gene cassettes of L. hongkongensis using the conjugation-mediated gene deletion system and examining their effects in acidic environment in vitro, in macrophages and in a mouse model. Results HLHK9ΔureA, HLHK9ΔureC, HLHK9ΔureD and HLHK9ΔureE all exhibited no urease activity. HLHK9ΔarcA1 and HLHK9ΔarcA2 both exhibited arginine deiminase (ADI) activities, but HLHK9ΔarcA1/arcA2 double deletion mutant exhibited no ADI activity. At pH 2 and 3, survival of HLHK9ΔarcA1/arcA2 and HLHK9ΔureA/arcA1/arcA2 were markedly decreased (p < 0.001) but that of HLHK9ΔureA was slightly decreased (p < 0.05), compared to wild type L. hongkongensis HLHK9. Survival of HLHK9ΔureA/arcA1/arcA2 and HLHK9ΔarcA1/arcA2 in macrophages were also markedly decreased (p < 0.001 and p < 0.01 respectively) but that of HLHK9ΔureA was slightly decreased (p < 0.05), compared to HLHK9, although expression of arcA1, arcA2 and ureA genes were all upregulated. Using a mouse model, HLHK9ΔureA exhibited similar survival compared to HLHK9 after passing through the murine stomach, but survival of HLHK9ΔarcA1/arcA2 and HLHK9ΔureA/arcA1/arcA2 were markedly reduced (p < 0.01). Conclusions In contrast to other important gastrointestinal tract pathogens, ADI pathway is far more important than urease for acid resistance and intracellular survival in L. hongkongensis. The gene duplication of the arc gene cassettes could be a result of their functional importance in L. hongkongensis. [ABSTRACT FROM AUTHOR]