Background: Research on mosquito-microbe interactions may lead to new tools for mosquito and mosquito-borne disease control. To date, such research has largely utilized laboratory-reared mosquitoes that typically lack the microbial diversity of wild populations. A logical progression in this area involves working under controlled settings using field-collected mosquitoes or, in most cases, their progeny. Thus, an understanding of how laboratory colonization affects the assemblage of mosquito microbiota would aid in advancing mosquito microbiome studies and their applications beyond laboratory settings. Methods: Using high throughput 16S rRNA amplicon sequencing, we characterized the internal and cuticle surface microbiota of F1 progeny of wild-caught adult Anopheles albimanus from four locations in Guatemala. A total of 132 late instar larvae and 135 2-5day old, non-blood-fed virgin adult females that were reared under identical laboratory conditions, were pooled (3 individuals/pool) and analyzed. Results: Results showed geographical heterogeneity in both F1 larval internal (p=0.001; pseudo-F = 9.53) and cuticle surface (p=0.001; pseudo-F = 8.51) microbiota, and only F1 adult cuticle surface (p=0.001; pseudo-F = 4.5) microbiota, with a more homogenous adult internal microbiota (p=0.12; pseudo-F = 1.6) across collection sites. Overall, ASVs assigned to Leucobacter, Thorsellia, Chryseobacterium and uncharacterized Enterobacteriaceae, dominated F1 larval internal microbiota, while Acidovorax, Paucibacter, and uncharacterized Comamonadaceae, dominated the larval cuticle surface. F1 adults comprised a less diverse microbiota compared to larvae, with ASVs assigned to the genus Asaia dominating both internal and cuticle surface microbiota, and constituting at least 70% of taxa in each microbial niche. Conclusions: These results suggest that location-specific heterogeneity in filed mosquito microbiota can be transferred to F1 progeny under normal laboratory conditions, but this may not last beyond the F1 larval stage without adjustments to maintain field-derived microbiota. Our findings provide the first comprehensive characterization of laboratory-colonized F1 An. albimanus progeny from field-derived mothers. This provides a background for studying how parentage and environmental conditions differentially or concomitantly affect mosquito microbiome composition, and how this can be exploited in advancing mosquito microbiome studies and their applications beyond laboratory settings.

中文翻译：

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实验室饲养的F1按蚊不同来源的内部和表皮表面微生物群的全面表征

背景：对蚊虫-微生物相互作用的研究可能会导致新的蚊虫和蚊媒疾病控制工具。迄今为止，此类研究已在很大程度上利用了实验室饲养的蚊子，这些蚊子通常缺乏野生种群的微生物多样性。在这一领域中，逻辑上的进展包括使用田间采集的蚊子，或者在大多数情况下，是它们的后代，在受控的条件下进行工作。因此，了解实验室定植如何影响蚊帐菌群的组装将有助于推进蚊帐微生物组研究及其在实验室环境之外的应用。方法：使用高通量16S rRNA扩增子序列，我们从危地马拉的四个地点表征了野生捕获的成年按蚊F1子代的内部和表皮表面微生物区系。汇集了在相同实验室条件下饲养的132只晚龄幼虫和135只2-5日龄，非血液喂养的成年雌性，进行了分析（每池3个人）并进行了分析。结果：结果显示F1幼虫内部（p = 0.001；假F = 9.53）和表皮表面（p = 0.001； pseudo-F = 8.51）微生物群和仅F1成年表皮表面（p = 0.001；假）均存在地理异质性-F = 4.5）菌群，跨收集点具有更均一的成年内部菌群（p = 0.12;假F = 1.6）。总体而言，分配给白杆菌，Thorsellia，Chryseobacterium和未表征的肠杆菌科的ASV占F1幼虫内部微生物群的主导，而Acidovorax，Paucibacter和未表征的Comamonadaceae占幼虫表皮表面的主导。与幼虫相比，F1成虫的微生物群落种类较少，分配给Asaia属的ASV占据了内部和表皮表面微生物群，并且在每个微生物生态位中至少构成了类群的70％。结论：这些结果表明，在正常实验室条件下，已归档蚊子菌群中特定于位置的异质性可以转移到F1子代中，但是如果不进行调整以保持田间微生物群，这种情况可能不会持续到F1幼虫期之后。我们的发现提供了实验室定植的F1 An的第一个全面表征。来自野外母亲的albimanus后代。这为研究育儿和环境条件如何差异或同时影响蚊子微生物组组成，以及如何将其用于推进蚊子微生物组研究及其在实验室环境之外的应用提供了背景。