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Vibrio alginolyticus, an opportunistic pathogen that causes vibriosis in miscellaneous fish species, has brought about serious economic damage to the mariculture industry in South China. The mechanism of virulence regulation in V. alginolyticus is yet not known except a Vibrio harveyi-like quorum sensing (QS) system that is established to manipulate the expression of diverse genes including those encoding virulence determinants. In this study, a new TetR family QS regulator, luxT, was identified and characterized in V. alginolyticus. The transcription of luxT gene was cell density dependent and was positively regulated by LuxU, an established QS component relaying the signal from three paralleled QS regulatory systems in V. harveyi. In addition, luxT positively regulated both luxO at transcriptional level and luxR at post-transcriptional level, which is thoroughly different from the established QS regulation mode in V. harveyi and Vibrio vulnificus. The mutant of luxT deletion produced markedly decreased total extracellular proteases and reduced motility ability compared to the wild type and the complemented strain luxT. The fish infection results indicated that mutation of luxT led to marginal attenuation in the virulence of V. alginolyticus, suggesting that LuxT might play a role in the fine-tuning of the virulence via QS in V. alginolyticus. [ABSTRACT FROM AUTHOR]

Abstract: The fish pathogen Vibrio alginolyticus virulence factors and regulation mechanism are barely known except the quorum sensing regulated extracellular products. In the present study, the cleavable isotope-coded affinity tags (cICATs) were used to analyze the differentially expressed extracellular proteins of V. alginolyticus wild-type and a ΔluxO mutant, and a putative peptidase Pep was identified. Quantitative real-time reverse transcription PCR confirmed that the transcription of pep was down-regulated by LuxO. Furthermore, western blotting demonstrated that the expression of Pep was cell-density dependent and quorum sensing regulated in V. alginolyticus. The pep in-frame mutant abolished the swimming and swarming motility, suggesting that it was involved in motility regulation in V. alginolyticus. In liquid medium, the polar flagellum was observed intact in ∆pep mutant while the chemotaxis related gene cheY, which is a response regulator causing change in direction of flagellar rotation, was down-regulated in ∆pep mutant. When ∆pep mutant was grown on hard agar plate, the lateral flagella disappeared and the transcriptions of flagellar biogenesis genes flaK, fliS, lafK, and lafA were down-regulated in ∆pep mutant. These results suggested that Pep was a luxO-regulated extracellular protein and involved in the motility of V. alginolyticus. [Copyright &y& Elsevier]

In Gram-negative bacteria, the TonB/ExbB/ExbD complex is required to energize the specific high-affinity receptors mediating iron uptake processes. In fish pathogen Vibrio alginolyticus MVP01, which exhibited capacities to scavenge iron sources with various specific iron uptake systems, we identified and characterized two sets of TonB systems. In V. alginolyticus, TonB1 and TonB2 systems were arrayed as TonB1–ExbB1–ExbD1 and ExbB2–ExbD2–TonB2, respectively. The transcription of the TonB systems was regulated by the iron concentration in culture medium. While TonB1 system specifically contributed to the hemin and hemoglobin utilization in V. alginolyticus MVP01, both of the TonB systems showed abilities to support the utilization of iron sources from ferrichrome and vibrioferrin, the endogenous siderophore of V. alginolyticus. In addition, when mutants of the TonB systems were inoculated intraperitoneally into the fish Brachydanio rerio, marked attenuation in virulence of V. alginolyticus was observed, indicating that both of the TonB systems are essential for the virulence of V. alginolyticus. [ABSTRACT FROM AUTHOR]

Vibrio alginolyticus, a marine bacterium, is an opportunistic pathogen capable of causing vibriosis with high mortality to fishes in the South China Sea. Stress resistance is very important for its survival in the natural environment and upon infection of the host. RpoS, an alternative sigma factor, is considered as an important regulator involved in stress response and virulence in many pathogens. In this study, the rpoS gene was cloned and characterized to evaluate the role of RpoS in V. alginolyticus. The predicted protein showed high identity with other reported rpoS gene products. The in-frame deleted mutation of rpoS in V. alginolyticus led to sensitivity of the strain to ethanol, hyperosmolarity, heat, and hydrogen peroxide challenges. Further studies showed that extracellular autoinducer 2 level, four of seven detected protease activities, and cytotoxicity of extracellular products were markedly decreased in the rpoS mutant compared with that in the wild-type strain. The results indicated that the global regulator RpoS was part of the regulatory networks of virulence and LuxS quorum sensing system. [ABSTRACT FROM AUTHOR]