Bacterial virulence analysis using brine shrimp as an infection model in relation to the importance of quorum sensing and proteases
Bacterial virulence analysis using brine shrimp as an infection model in relation to the importance of quorum sensing and proteases
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DOI：
10.2323/jgam.60.169
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发表时间：
2014-01-01
影响因子：
1.2
通讯作者：
Lee, Joon-Hee
中科院分区：
文献类型：
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作者：
Lee, Mi-Nan;Kim, Soo-Kyoung;Lee, Joon-Hee
卤虫是一种水生甲壳类动物，属于卤虫属。这种微生物被广泛用于测试化学品的毒性。本研究以卤虫为感染模式生物，研究细菌的毒力。用创伤弧菌、铜绿假单胞菌、越南伯克霍尔德氏菌、金黄色葡萄球菌和大肠杆菌等多种病原菌感染卤虫无节幼体，通过计数感染无节幼体的存活率来研究其对这些细菌的敏感性。所有供试细菌对卤虫均具有明显的毒力，几天内即可杀死卤虫无节幼体，其中创伤弧菌的毒力最强。铜绿假单胞菌对卤虫的毒力也表现出剂量依赖性，但毒力弱于创伤弧菌。利用创伤弧菌和铜绿假单胞菌的毒力减弱突变体（如群体感应（QS）突变体或蛋白酶缺陷突变体）进行的毒力测试显示毒力显著减弱，表明QS机制在这些细菌对卤虫的毒力中是重要的。B。vietnamiensis、S.金黄色葡萄球菌和E.大肠杆菌对卤虫也有一定的毒力，且毒力在24 h内与投加剂量有关。沙门氏菌和枯草芽孢杆菌对卤虫也有一定的毒力，但与其他细菌相比，其毒力较弱，且发挥缓慢。因此，卤虫是一种较好的细菌毒力测定模型，尤其是创伤弧菌和铜绿假单胞菌，QS在卤虫致病力测定中具有重要作用。
Brine shrimp are aquatic crustaceans belonging to a genus of Artemia. This organism is widely used for testing the toxicity of chemicals. In this study, brine shrimp were evaluated as an infection model organism to study bacterial virulence. Artemia nauplii were infected with various pathogenic bacteria, such as Vibrio vulnificus, Pseudomonas aeruginosa, Burkholderia vietnamiensis, Staphylococcus aureus, and Escherichia coli, and the susceptibility to these bacteria was investigated by counting the survival of the infected nauplii. While all of the tested bacteria have significant virulence to brine shrimp, killing the nauplii in a few days, V. vulnificus showed the strongest virulence. P. aeruginosa also showed a dose-dependent virulence to brine shrimp, but the virulence was weaker than that of V. vulnificus. The virulence tests using the virulence-attenuated mutants of V. vulnificus and P. aeruginosa, such as quorum sensing (QS) mutants or protease-deficient mutants showed a significant attenuation of virulence, demonstrating that the QS mechanism is important in the virulence of these bacteria to brine shrimp. B. vietnamiensis, S. aureus, and E. coli were also virulent to brine shrimp and the virulence was correlated with dosage within 24 h under our conditions. Salmonella enterica Typhimurium and Bacillus subtilis were also virulent to brine shrimp, but the virulence was weak and slowly exerted compared with that of other bacteria. Taken together, we suggest that brine shrimp are a good infection model to assay bacterial virulence, especially for V. vulnificus and P. aeruginosa, and QS is important in the bacterial virulence to brine shrimp.