W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA.
Sci Rep. 2018 May 29;8(1):8358. doi: 10.1038/s41598-018-26680-2.
Mosquito larvae continuously encounter microbes in their aquatic environment, which serve as food and play a critical role in successful development. In previous work, we isolated a Chromobacterium sp. (C.sp_P) with larvicidal activity from the midgut of dengue vector Aedes mosquitoes in Panama. In this study, we found a positive correlation between initial concentrations of C.sp_P and larval mortality rates, and that C.sp_P is more efficient at inducing larval mortality in a high nutrient environment. Multiple Chromobacterium species induce larval mortality with similar efficacy to C.sp_P except for C. subtsugae. We also found that a non-lethal dose of C.sp_P lengthens development time and increases mortality over multiple developmental stages, suggesting persistent effects of exposure. Additionally, we showed that larvicidal activity persists in the larval breeding water after removal of live bacteria, and that the larvicidal factor in C.sp_P-treated water is smaller than 3 kDa, heat resistant to 90 °C, and lost after vacuum centrifugation. We showed that C.sp_P produces hydrogen cyanide in culture and in larval water at concentrations sufficient to kill An. gambiae larvae, and treatment of the larval water with a cyanide antidote eliminated larvicidal activity. We conclude that a potential mechanism by which C.sp_P can induce larval mortality is via production of hydrogen cyanide.
蚊子幼虫在水生环境中不断接触微生物,这些微生物既是食物来源,又对其成功发育起着关键作用。在之前的工作中,我们从巴拿马登革热病媒埃及伊蚊的中肠中分离出一种杀蚊幼虫的噬色菌(Chromobacterium sp.,C.sp_P)。在本研究中,我们发现 C.sp_P 的初始浓度与幼虫死亡率之间呈正相关,并且 C.sp_P 在高营养环境中更能有效诱导幼虫死亡。除了 C. subtsugae 外,多种噬色菌诱导幼虫死亡的效果与 C.sp_P 相似。我们还发现,C.sp_P 的非致死剂量会延长发育时间,并在多个发育阶段增加死亡率,表明其暴露后具有持续效应。此外,我们表明,在去除活细菌后,C.sp_P 处理过的水中仍保留杀幼虫活性,并且 C.sp_P 处理水中的杀幼虫因子小于 3 kDa,对 90°C 的热具有抗性,并且在真空离心后失去活性。我们表明,C.sp_P 在培养物中和幼虫水中产生足以杀死 An. gambiae 幼虫的氰化氢,并且用氰化物解毒剂处理幼虫水会消除杀幼虫活性。我们得出结论,C.sp_P 诱导幼虫死亡的潜在机制可能是通过产生氰化氢。