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1. A cryopreservation method to recover laboratory- and field-derived bacterial communities from mosquito larval habitats

   https://doi.org/10.1371/journal.pntd.0011234  

   Zhao, Serena Y.; Hughes, Grant L.; Coon, Kerri L. (April 2023, PLOS Neglected Tropical Diseases)

   Bartholomay, Lyric C. (Ed.)

   Mosquitoes develop in a wide range of aquatic habitats containing highly diverse and variable bacterial communities that shape both larval and adult traits, including the capacity of adult females of some mosquito species to transmit disease-causing organisms to humans. However, while most mosquito studies control for host genotype and environmental conditions, the impact of microbiota variation on phenotypic outcomes of mosquitoes is often unaccounted for. The inability to conduct reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions has also greatly limited our ability to identify microbial targets for mosquito-borne disease control. Here, we developed an approach to isolate and cryopreserve bacterial communities derived from lab and field-based larval rearing environments of the yellow fever mosquito Aedes aegypti –a primary vector of dengue, Zika, and chikungunya viruses. We then validated the use of our approach to generate experimental microcosms colonized by standardized lab- and field-derived bacterial communities. Our results overall reveal minimal effects of cryopreservation on the recovery of both lab- and field-derived bacteria when directly compared with isolation from non-cryopreserved fresh material. Our results also reveal improved reproducibility of bacterial communities in replicate microcosms generated using cryopreserved stocks over fresh material. Communities in replicate microcosms further captured the majority of total bacterial diversity present in both lab- and field-based larval environments, although the relative richness of recovered taxa as compared to non-recovered taxa was substantially lower in microcosms containing field-derived bacteria. Altogether, these results provide a critical next step toward the standardization of mosquito studies to include larval rearing environments colonized by defined microbial communities. They also lay the foundation for long-term studies of mosquito-microbe interactions and the identification and manipulation of taxa with potential to reduce mosquito vectorial capacity. 

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2. The microbiome and mosquito vectorial capacity: rich potential for discovery and translation

   https://doi.org/10.1186/s40168-021-01073-2  

   Cansado-Utrilla, Cintia; Zhao, Serena Y.; McCall, Philip J.; Coon, Kerri L.; Hughes, Grant L. (May 2021, Microbiome)

   Abstract Microbiome research has gained considerable interest due to the emerging evidence of its impact on human and animal health. As in other animals, the gut-associated microbiota of mosquitoes affect host fitness and other phenotypes. It is now well established that microbes can alter pathogen transmission in mosquitoes, either positively or negatively, and avenues are being explored to exploit microbes for vector control. However, less attention has been paid to how microbiota affect phenotypes that impact vectorial capacity. Several mosquito and pathogen components, such as vector density, biting rate, survival, vector competence, and the pathogen extrinsic incubation period all influence pathogen transmission. Recent studies also indicate that mosquito gut-associated microbes can impact each of these components, and therefore ultimately modulate vectorial capacity. Promisingly, this expands the options available to exploit microbes for vector control by also targeting parameters that affect vectorial capacity. However, there are still many knowledge gaps regarding  mosquito–microbe interactions  that need to be addressed in order to exploit them efficiently. Here, we review current evidence of impacts of the microbiome on aspects of vectorial capacity, and we highlight likely opportunities for novel vector control strategies and areas where further studies are required. 

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3. Abiotic factors shape mosquito microbiomes that enhance host development

   https://doi.org/10.1093/ismejo/wrae181  

   Kriefall, Nicola G; Seabourn, Priscilla S; Yoneishi, Nicole M; Davis, Kahiwahiwa; Nakayama, Kirsten K; Weber, Danya E; Hynson, Nicole A; Medeiros, Matthew_C I (January 2024, The ISME Journal)

   Abstract Metazoans rely on interactions with microorganisms through multiple life stages. For example, developmental trajectories of mosquitoes can vary depending on the microorganisms available during their aquatic larval phase. However, the role that the local environment plays in shaping such host-microbe dynamics and the consequences for the host organism remain inadequately understood. Here, we examine the influence of abiotic factors, locally available bacteria, and their interactions on the development and associated microbiota of the mosquito Aedes albopictus. Our findings reveal that leaf detritus infused into the larval habitat water, sourced from native Hawaiian tree ‘ōhi‘a lehua Metrosideros polymorpha, invasive strawberry guava Psidium cattleianum, or a pure water control, displayed a more substantial influence than either temperature variations or simulated microbial dispersal regimes on bacterial community composition in adult mosquitoes. However, specific bacteria exhibited divergent patterns within mosquitoes across detrital infusions that did not align with their abundance in the larval habitat. Specifically, we observed a higher relative abundance of a Chryseobacterium sp. strain in mosquitoes from the strawberry guava infusion than the pure water control, whereas the opposite trend was observed for a Pseudomonas sp. strain. In a follow-up experiment, we manipulated the presence of these two bacterial strains and found larval developmental success was enhanced by including the Chryseobacterium sp. strain in the strawberry guava infusion and the Pseudomonas sp. strain in the pure water control. Collectively, these data suggest that interactions between abiotic factors and microbes of the larval environment can help shape mosquito populations' success. 

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4. Microbial communities of container aquatic habitats shift in response to Culex restuans larvae

   https://doi.org/10.1093/femsec/fiaa112  

   Muturi, Ephantus J; Dunlap, Christopher; Cáceres, Carla E (July 2020, FEMS Microbiology Ecology)

   ABSTRACT We examined how larvae of Culex restuans mosquito influences the bacterial abundance, composition and diversity in simulated container aquatic habitats. The microbiota of Cx. restuans larvae were also characterized and compared to those of their larval habitats. The presence of Cx. restuans larvae altered the bacterial community composition and reduced the bacterial abundance, diversity and richness. Azohydromonas sp., Delftia sp., Pseudomonas sp., Zooglea sp., unclassified Enterobacteriaceae and unclassified Bacteroidales were suppressed while Prosthecobacter sp., Hydrogenaphaga sp., Clostridium sp., unclassified Clostridiaceae and Chryseobacterium sp. were enhanced in the presence of Cx. restuans larvae. Cx. restuans larvae harbored distinct and less diverse bacterial community compared to their larval habitats. These findings demonstrate that Cx. restuans larvae play a key role in structuring the microbial communities in container aquatic habitats and may lower the nutritional quality and alter the decomposition process and food web dynamics in these aquatic systems. The findings also demonstrate that mosquito larvae are highly selective of the bacterial taxa from the larval environment that colonize their bodies. These findings provide new opportunities for more focused studies to identify the specific bacterial taxa that serve as food for mosquito larvae and those that could be harnessed for disease control. 

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5. Interspecies microbiome transplantation recapitulates microbial acquisition in mosquitoes

   https://doi.org/10.1186/s40168-022-01256-5  

   Coon, Kerri L.; Hegde, Shivanand; Hughes, Grant L. (April 2022, Microbiome)

   Abstract BackgroundMosquitoes harbor microbial communities that play important roles in their growth, survival, reproduction, and ability to transmit human pathogens. Microbiome transplantation approaches are often used to study host-microbe interactions and identify microbial taxa and assemblages associated with health or disease. However, no such approaches have been developed to manipulate the microbiota of mosquitoes. ResultsHere, we developed an approach to transfer entire microbial communities between mosquito cohorts. We undertook transfers between (Culex quinquefasciatustoAedes aegypti) and within (Ae. aegyptitoAe. aegypti) species to validate the approach and determine the number of mosquitoes required to prepare donor microbiota. After the transfer, we monitored mosquito development and microbiota dynamics throughout the life cycle. Typical holometabolous lifestyle-related microbiota structures were observed, with higher dynamics of microbial structures in larval stages, including the larval water, and less diversity in adults. Microbiota diversity in recipient adults was also more similar to the microbiota diversity in donor adults. ConclusionsThis study provides the first evidence for successful microbiome transplantation in mosquitoes. Our results highlight the value of such methods for studying mosquito-microbe interactions and lay the foundation for future studies to elucidate the factors underlying microbiota acquisition, assembly, and function in mosquitoes under controlled conditions. 

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