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1. Culex pipiens and Culex restuans egg rafts harbor diverse bacterial communities compared to their midgut tissues

   https://doi.org/10.1186/s13071-020-04408-4  

   Juma, Elijah O. ; Kim, Chang-Hyun ; Dunlap, Christopher ; Allan, Brian F. ; Stone, Chris M. ( December 2020 , Parasites & Vectors)

   null (Ed.)

   Abstract Background The bacterial communities associated with mosquito eggs are an essential component of the mosquito microbiota, yet there are few studies characterizing and comparing the microbiota of mosquito eggs to other host tissues. Methods We sampled gravid female Culex pipiens L. and Culex restuans Theobald from the field, allowed them to oviposit in the laboratory, and characterized the bacterial communities associated with their egg rafts and midguts for comparison through MiSeq sequencing of the 16S rRNA gene. Results Bacterial richness was higher in egg rafts than in midguts for both species, and higher in Cx pipiens than Cx. restuans . The midgut samples of Cx. pipiens and Cx. restuans were dominated by Providencia. Culex pipiens and Cx. restuans egg rafts samples were dominated by Ralstonia and Novosphingobium , respectively. NMDS ordination based on Bray-Curtis distance matrix revealed that egg-raft samples, or midgut tissues harbored similar bacterial communities regardless of the mosquito species. Within each mosquito species, there was a distinct clustering of bacterial communities between egg raft and midgut tissues. Conclusion These findings expand the list of described bacterial communities associated with Cx. pipiens and Cx. restuan s and the additional characterization of the egg raft bacterial communities facilitates comparative analysis of mosquito host tissues, providing a basis for future studies seeking to understand any functional role of the bacterial communities in mosquito biology. 

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2. Effect of life stage and pesticide exposure on the gut microbiota of Aedes albopictus and Culex pipiens L

   https://doi.org/10.1038/s41598-020-66452-5  

   Juma, Elijah O. ; Allan, Brian F. ; Kim, Chang-Hyun ; Stone, Christopher ; Dunlap, Christopher ; Muturi, Ephantus J. ( December 2020 , Scientific Reports)
3. Condition-Specific Competitive Effects of the Invasive Mosquito Aedes albopictus on the Resident Culex pipiens among Different Urban Container Habitats May Explain Their Coexistence in the Field

   https://doi.org/10.3390/insects12110993  

   Leisnham, Paul T. ; LaDeau, Shannon L. ; Saunders, Megan E. ; Villena, Oswaldo C. ( November 2021 , Insects)

   null (Ed.)

   Condition-specific competition, when environmental conditions alter the outcome of competition, can foster the persistence of resident species after the invasion of a competitively superior invader. We test whether condition-specific competition can facilitate the areawide persistence of the resident and principal West Nile virus vector mosquito Culex pipiens with the competitively superior invasive Aedes albopictus in water from different urban container habitats. (2) Methods: We tested the effects of manipulated numbers of A. albopictus on C. pipiens’ survival and development in water collected from common functional and discarded containers in Baltimore, MD, USA. The experiment was conducted with typical numbers of larvae found in field surveys of C. pipiens and A. albopictus and container water quality. (3) Results: We found increased densities of A. albopictus negatively affected the survivorship and development of C. pipiens in water from discarded containers but had little effect in water from functional containers. This finding was driven by water from trash cans, which allowed consistently higher C. pipiens’ survival and development and had greater mean ammonia and nitrate concentrations that can promote microbial food than other container types. (4) Conclusions: These results suggest that the contents of different urban containers alter the effects of invasive A. albopictus competition on resident C. pipiens, that trash cans, in particular, facilitate the persistence of C. pipiens, and that there could be implications for West Nile virus risk as a result. 

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4. Light Pollution Disrupts Seasonal Differences in the Daily Activity and Metabolic Profiles of the Northern House Mosquito, Culex pipiens

   Wolkoff, Matthew ; Fyie, Lydia ; Meuti, Megan E. ( January 2023 , Insects)

   The Northern house mosquito transmits West Nile virus and survives the winter by entering a state of dormancy called diapause. Light pollution has been shown to interfere with diapause initiation in this mosquito. The effects of light pollution on daily activity rhythms and metabolic products have not been thoroughly investigated in diapausing and non-diapausing Northern house mosquitoes. We found that light pollution affected mosquito activity levels and several metabolic products differently depending on photoperiod, indicating that light pollution may disrupt nutrient accumulation and possibly interfere with diapause initiation in this species. 

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5. ReMOT Control Delivery of CRISPR-Cas9 Ribonucleoprotein Complex to Induce Germline Mutagenesis in the Disease Vector Mosquitoes Culex pipiens pallens (Diptera: Culicidae)

   https://doi.org/10.1093/jme/tjab016  

   Li, Xixi ; Xu, Yang ; Zhang, Hongbo ; Yin, Haitao ; Zhou, Dan ; Sun, Yan ; Ma, Lei ; Shen, Bo ; Zhu, Changliang ( February 2021 , Journal of Medical Entomology)

   Slotman, Michel (Ed.)

   Abstract The wide distribution of Culex (Cx.) pipiens complex mosquitoes makes it difficult to prevent the transmission of mosquito-borne diseases in humans. Gene editing using CRISPR/Cas9 is an effective technique with the potential to solve the growing problem of mosquito-borne diseases. This study uses the ReMOT Control technique in Culex pipiens pallens (L.) to produce genetically modified mosquitoes. A microinjection system was established by injecting 60 adult female mosquitoes—14 µl injection mixture was required, and no precipitation occurred with ≤1 µl of endosomal release reagents (chloroquine or saponin). The efficiency of delivery of the P2C-enhanced green fluorescent protein-Cas9 (P2C-EGFP-Cas9) ribonucleoprotein complex into the ovary was 100% when injected at 24 h post-bloodmeal (the peak of vitellogenesis). Using this method for KMO knockout, we found that gene editing in the ovary could also occur when P2C-Cas9 RNP complex was injected into the hemolymph of adult Cx. pipiens pallens by ReMOT Control. In the chloroquine group, of the 2,251 G0 progeny screened, 9 individuals showed with white and mosaic eye phenotypes. In the saponin group, of the 2,462 G0 progeny screened, 8 mutant individuals were observed. Sequencing results showed 13 bp deletions, further confirming the fact that gene editing occurred. In conclusion, the successful application of ReMOT Control in Cx. pipiens pallens not only provides the basic parameters (injection parameters and injection time) for this method but also facilitates the study of mosquito biology and control. 

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