Metagenome Sequencing Reveals the Midgut Microbiota Makeup of Culex pipiens quinquefasciatus and Its Possible Relationship With Insecticide Resistance

Midgut microbiota can participate in the detoxification and metabolism processes in insects, but there are few reports on the relationship between midgut microbiota and insecticide resistance in mosquitoes. In this study, we performed metagenomic sequencing on a susceptible strain (SS), a field-collected Hainan strain (HN), and a deltamethrin-resistant strain (RR) of Culex pipiens quinquefasciatus to understand the diversity and functions of their midgut microbiota. The results revealed differences in midgut microbiota among the three strains of Cx. pipiens quinquefasciatus. At the phylum level, Proteobacteria was the most prominent, accounting for nearly 70% of their midgut microbes. At the genus level, Aeromonas made up the highest proportion. In addition, Aeromonas, Morganella, Elizabethkingia, Enterobacter, Cedecea, and Thorsellia showed significant differences between strains. At the species level, Bacillus cereus, Enterobacter cloacae complex sp. 4DZ3-17B2, Streptomyces sp. CNQ329, and some species of Pseudomonas and Wolbachia were more abundant in the two resistant strains. Principal component analysis (PCA) showed that the SS strain had significantly different metagenomic functions than the two deltamethrin-resistant strains (HN and RR strain). The HN and RR strains differed from the SS strain in more than 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The analysis of species abundance and functional diversity can provide directions for future studies.

Transcript Assembly and Quantification by RNA-Seq Reveals Significant Differences in Gene Expression and Genetic Variants in Mosquitoes of the Culex pipiens (Diptera: Culicidae) Complex

The taxonomy of Culex pipiens complex of mosquitoes is still debated, but in North America it is generally regarded to include Culex pipiens pipiens, Culex pipiens molestus, and Culex quinquefasciatus (or Culex pipiens quinquefasciatus). Although these mosquitoes have very similar morphometry, they each have unique life strategies specifically adapted to their ecological niche. Differences include the capability for overwintering diapause, bloodmeal preference, mating behaviors, and reliance on blood meals to produce eggs. Here, we used RNA-seq transcriptome analysis to investigate the differential gene expression and nucleotide polymorphisms that may link to the divergent traits specifically between Cx. pipiens pipiens and Cx. pipiens molestus.

Culex interfor and Culex saltanensis (Diptera: Culicidae) are susceptible and competent to transmit St. Louis encephalitis virus (Flavivirus: Flaviviridae) in central Argentina

Background St. Louis encephalitis virus (SLEV) is endemic and autochthonous on the American continent. Culex pipiens quinquefasciatus is a vector of SLEV; however, Culex interfor and Culex saltanensis have also been found to be naturally infected with SLEV. The aim of this study was to determine the vector competence of C. interfor and C. saltanensis for SLEV from Argentina compared with C. p. quinquefasciatus. Methods Female of the Culex species were orally infected by feeding on viraemic chicks that had been inoculated with SLEV. Abdomens, legs and saliva blood-fed mosquitoes were analysed by viral plaque assay. Results Mosquitoes were susceptible to orally acquired infection, dissemination and transmission of SLEV in the saliva. Conclusions Our results demonstrate that C. saltanensis and C. interfor are susceptible to SLEV and competent for its transmission.

Effects of Propoxur Exposure on Insecticidal Susceptibility and Developmental Traits in Culex pipiens quinquefasciatus

Propoxur-sel strains of Culex pipiens quinquefasciatus were derived from a lab-bred strain following 16 generations of propoxur exposure under sublethal concentrations of LC25 (lethal concentration of 25%) and LC50 (lethal concentration of 50%), respectively. This resulted in resistance development in F16 with ratios of 8.8× and 6.3×, respectively, compared with F0. The fecundity, longevity, sex ratio (F/M), and hatchability of the propoxur-exposed Cx. quinquefasciatus adult survivors and their offspring were decreased, with no effect on the emergence ratio and pupa survival rate. In addition, the intrinsic rates of increase (r), the net reproduction (R0), and the finite rate of increase (λ) of the Cx. quinquefasciatus offspring generations were also decreased significantly compared to F0. Correspondingly, the mean generation time (T) and the population double time (DT) in propoxur-sels were increased. Enhanced activities of cytochrome P450 monooxygenase and esterase were also observed in propoxur-sels, indicating that a detoxification mechanism might be responsible for resistance development in Cx. quinquefasciatus. Except for the three genes cyp4d42v1, cyp4c52v1, and cyp6aa9 which displayed a coincidence in some degree in different treatments, induction by different doses of propoxur and constitutive expression in different generations of propoxur-sel strains resulted in an inconsistent identification of the P450 genes probably related with resistance.

Culex saltanensis and Culex interfor (Diptera: Culicidae) are susceptible and competent to transmit St. Louis encephalitis virus (Flavivirus: Flaviviridae) in central Argentina.

Infectious diseases caused by mosquito-borne viruses constitute health and economic problems worldwide. St. Louis encephalitis virus (SLEV) is endemic and autochthonous in the American continent. Culex pipiens quinquefasciatus is the primary urban vector of SLEV; however, Culex interfor and Culex saltanensis have also been found naturally infected with the virus, suggesting their potential role as vectors. OBJECTIVE: The aim of this study was to determine the vector competence of Cx. interfor and Cx. saltanensis for SLEV from central Argentina in comparison to Cx. p. quinquefasciatus. METHODS: Adult female mosquitoes of the three Culex species were orally infected by feeding on viremic chicks that had been inoculated with SLEV. Then, abdomens, legs and saliva blood-fed mosquitoes were analyzed by viral plaque assay and the presence of cytopathic effect on the cell culture monolayer. RESULTS: Mosquitoes were permissive to orally acquired infections, to virus dissemination, and transmission of SLEV in the saliva. Cx. saltanensis and Cx. interfor are potential vectors of SLEV. CONCLUSIONS: Our results demonstrate that in Argentina both Cx. saltanensis and Cx. interfor are susceptible to SLEV and competent for its transmission. Moreover they are abundant during SLEV epidemic period in urban area, positive for this virus in nature, and found to feed on natural hosts.