Abstract 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 F 1 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 F 1 larval internal ( p =0.001; pseudo- F = 9.53) and cuticle surface ( p =0.001; pseudo- F = 8.51) microbiota, and only F 1 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 F 1 larval internal microbiota, while Acidovorax, Paucibacter , and uncharacterized Comamonadaceae , dominated the larval cuticle surface. F 1 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 F 1 progeny under normal laboratory conditions, but this may not last beyond the F 1 larval stage without adjustments to maintain field-derived microbiota. Our findings provide the first comprehensive characterization of laboratory-colonized F 1 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.

Comprehensive characterization of internal and cuticle surface microbiota of laboratory-reared F<sub>1</sub><i>Anopheles albimanus</i> originating from different sites

Comprehensive characterization of internal and cuticle surface microbiota of laboratory-reared F1Anopheles albimanus originating from different sites