Comprehensive characterization of internal and cuticle surface microbiota of laboratory-reared F1 Anopheles albimanus originating from different sites

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, the internal and cuticle surface microbiota of F1 progeny of wild-caught adult Anopheles albimanus from four locations in Guatemala were characterized. A total of 132 late instar larvae and 135 2–5 day-old, non-blood-fed virgin adult females that were reared under identical laboratory conditions, were pooled (3 individuals/pool) and analysed. Results Results showed location-associated 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. These findings provide the first comprehensive characterization of laboratory-colonized F1An. 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.

Microbiome structure of a wildDrosophilacommunity along tropical elevational gradients and comparison to laboratory lines

“…The substantial differences in microbiomes between lab and field specimens suggests that future studies should be cautious and specific with the types of microbiome-related questions investigating laboratory flies. Interpreting microbiome community composition from lab-kept specimens, particularly those from cultures maintained across multiple generations, is unlikely to yield data entirely representative of natural microbiomes 70,71 , except perhaps in scenarios where microbiome communities from lab-reared specimens are a subset of the more diverse microbiomes found in the field (as our results show in Figure 5).…”

Microbiome Structure of a Wild Drosophila Community along Tropical Elevational Gradients and Comparison to Laboratory Lines

“…The substantial differences in microbiomes between lab and field specimens suggest that future studies should be cautious and specific with the types of microbiome-related questions investigating laboratory flies. Interpreting microbiome community composition from lab-kept specimens, particularly those from cultures maintained across multiple generations, is unlikely to yield data entirely representative of natural microbiomes ( 58 , 59 ), except perhaps in scenarios where microbiome communities from lab-reared specimens are a subset of the more diverse microbiomes found in the field (NMDS mean stress ≈ 0.15, PERMANOVA, R 2 = 0.150, Benjamini-Hochberg corrected P ≤ 0.001, beta-dispersion F = 126.8 and P ≤ 0.001) (Table S1 and Fig. 1 ).…”

Current Status of Metatranscriptomic and Related Studies in Hematophagous Disease-Transmitting Vectors

“…Of these, the vast majority correspond to metataxonomic analyses that have used either DNA metabarcoding or RNA shotgun sequencing to study the different components of the microbiome separately. The metabarcoding analyses have mainly focused on bacteria (e.g., Boissière et al 2012;Buck et al 2016;Coon et al 2016Coon et al , 2014Dada et al 2021aDada et al , 2019Díaz et al 2021;Dickson et al 2017;Duguma et al 2019;Gimonneau et al 2014;Hegde et al 2018;Mancini et al 2018;Muturi et al 2016;Osei-Poku et al 2012;Sharma et al 2014;Trzebny et al 2023;Villegas et al 2018;, but a couple have analyzed the fungal (Tawidian et al 2021) and eukaryotic (Belda et al 2017) components, and one metabarcoding study included both prokaryotes and eukaryotes (Thongsripong et al 2018).…”