Differential expression of urease genes and ureolytic activity of uropathogenic Escherichia coli and Pseudomonas aeruginosa isolates in different nutritional conditions.

Uropathogens have adaptation strategies to survive in the host urinary tract by efficiently utilizing and tolerating the urinary metabolites. Many uropathogens harbour the enzyme urease for the breakdown of urea and the enzymatic breakdown of urea increases the pH and facilitate the struvite crystallization. In this study, the differential urease activity of uropathogenic Escherichia coli and Pseudomonas aeruginosa strains was investigated under different nutritional conditions. The experiments included measurement of growth, pH, urease activity, NH4-N generation and urease gene (ureC) expression among the bacterial strains under different conditions. Further, the implications of urea breakdown on the struvite crystallization in vitro and biofilm formation were also assessed. The study included urease positive isolates and for comparison urease negative isolates were included. Compared to the urease negative strains the urease positive strains formed higher biofilms and motility. The urease positive P. aeruginosa showed significantly higher (p < 0.01) pH and urease activity (A557-A630) compared to E. coli under experimental conditions. Further, supplementation of glucose to the growth media significantly increased the urease activity in P. aeruginosa and in contrast, it was significantly lower in E. coli. The expression profile of urease gene (ureC) was significantly higher (p < 0.001) in P. aeruginosa compared to E. coli and was consistent with the biochemical results of the urease activity under the nutritional conditions. The differential urease activity under two nutritional conditions influenced the biogenic struvite crystallization. It correlated with the urease activity showing higher crystallization rate in P. aeruginosa compared to E. coli. The results highlight the differential urease activity in two common uropathogens under different nutritional conditions that may have significant role on the regulation of virulence, pathogenicity and in the kidney stone disease.

Carvone - a quorum sensing inhibitor blocks biofilm formation in Chromobacterium violaceum.

Carvone is a natural monoterpenoid and in this study it was tested for its role in attenuating quorum sensing (QS) controlled biofilm formation in Chromobacterium violaceum. It showed significant QS inhibition in terms of reduction in violacein at a concentration range of 60 to 70 µg/mL against C. violaceum ATCC 12472. At the same concentration, carvone also inhibited biofilm formation by more than 80%. The biofilm morphology of C. violaceum is unique with a well organised pattern of cell arrangement in a tight matrix. The same was evident in Scanning electron microscopy, however, carvone treatment not only showed reduction in biofilm density but also disruption of biofilm matrix. Interruption of biofilm formation was attributed to reduction in the exopolysaccharide production and swarming motility. Molecular investigations (RT-PCR) showed that the important genes involved in biofilm regulation such as pilS, pilR, pilB and pilT were downregulated significantly in the treatment groups.