Project description:BackgroundThe 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.MethodsWe 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.ResultsBacterial 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.ConclusionThese findings expand the list of described bacterial communities associated with Cx. pipiens and Cx. restuans 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.
Project description:Mechanistic evidence of widespread insecticide resistance among Illinois West Nile virus vectors (Culex pipiens and Culex restuans)
Project description:Container aquatic habitats support a specialized community of macroinvertebrates (e.g. mosquitoes) that feed on microbial communities associated with decaying organic matter. These aquatic habitats are often embedded within and around agricultural lands and are frequently exposed to pesticides. We used a microcosm approach to examine the single and combined effects of two herbicides (atrazine, glyphosate), and three insecticides (malathion, carbaryl, permethrin) on microbial communities of container aquatic habitats. MiSeq sequencing of the V4 region of both bacterial and archaeal 16S rRNA gene was used to characterize the microbial communities of indoor microcosms that were either exposed to each pesticide alone, a mix of herbicides, a mix of insecticides, or a mix of all five insecticides. Individual insecticides but not herbicides reduced the microbial diversity and richness and two insecticides, carbaryl and permethrin, also altered the microbial community structure. A mixture of herbicides had no effect on microbial diversity or structure but a mixture of insecticides or all five pesticides reduced microbial diversity and altered the community structure. These findings suggest that exposure of aquatic ecosystems to individual pesticides or their mixtures can disrupt aquatic microbial communities and there is need to decipher how these changes affect resident macroinvertebrate communities.
Project description:Mosquitoes host diverse microbial communities that influence many aspects of their biology including reproduction, digestion, and ability to transmit pathogens. Unraveling the composition, structure, and function of these microbiota can provide new opportunities for exploiting microbial function for mosquito-borne disease control.MiSeq® sequencing of 16S rRNA gene amplicons was used to characterize the microbiota of adult females of Culex pipiens L. and Cx. restuans Theobald collected from nine study sites in central Illinois.Out of 195 bacterial OTUs that were identified, 86 were shared between the two mosquito species while 16 and 93 OTUs were unique to Cx. pipiens and Cx. restuans, respectively. The composition and structure of microbial communities differed significantly between the two mosquito species with Cx. restuans hosting a more diverse bacterial community compared to Cx. pipiens. Wolbachia (OTU836919) was the dominant bacterial species in Cx. pipiens accounting for 91% of total microbiota while Sphingomonas (OTU817982) was the dominant bacterial species in Cx. restuans accounting for 31% of total microbiota. Only 3 and 6 OTUs occurred in over 60% of individuals in Cx. pipiens and Cx. restuans, respectively. There was little effect of study site on bacterial community structure of either mosquito species.These results suggest that the two mosquito species support distinct microbial communities that are sparsely distributed between individuals. These findings will allow investigations of the role of identified microbiota on the spatial and temporal heterogeneity in WNV transmission and their potential application in disease control.
Project description:Mosquitoes of the genus Culex are important vectors of a variety of arthropod-borne viral infections. In most of the northern parts of the USA, Cx. pipiens/restuans is the predominant representative of this genus. As vectors, they play a key role in the spreading of arboviruses and thus, knowledge of the population dynamic of mosquitoes is important to understand the disease ecology of these viruses. As poikilotherm animals, the vital rates of mosquitoes are highly dependent on ambient temperature, and also on precipitation. We present a compartmental model for the population dynamics of Cx. pipiens/restuans. The model is driven by temperature, precipitation, and daytime length (which can be calculated from the geographic latitude). For model evaluation, we used long-term mosquito capture data, which were averaged from multiple sites in Cook County, Illinois. The model fitted the observation data and was able to reproduce between-year differences in the abundance of the Cx. pipiens/restuans mosquitoes, as well as the different seasonal trends. Using this model, we evaluated the effectiveness of targeting different vital rates for mosquito control strategies. The final model is able to reproduce the weekly mean Cx. pipiens/restuans abundance for Cook County with a high accuracy, and over a long time period of 20 years.
Project description:Culex pipiens molestus and Cx. p. quinquefasciatus are the members of Culex pipiens Complex, but they display relatively large differences in behavior and physiological responses. We compared the genes of these mosquitoes to identify those that were differentially expressed in each subspecies. Such genes could play important roles in subspecies-specific blood feeding or oviposition behavior.
Project description:We investigated the bacterial diversity of microbial communities in water-filled, human-made and natural container habitats of the mosquitoes Aedes aegypti and Aedes albopictus in suburban landscapes of New Orleans, Louisiana in 2003. We collected water samples from three classes of containers, including tires (n = 12), cemetery urns (n = 23), and miscellaneous containers that included two tree holes (n = 19). Total genomic DNA was extracted from water samples, and 16S ribosomal DNA fragments (operational taxonomic units, OTUs) were amplified by PCR and separated by denaturing gradient gel electrophoresis (DGGE). The bacterial communities in containers represented diverse DGGE-DNA banding patterns that were not related to the class of container or to the local spatial distribution of containers. Mean richness and evenness of OTUs were highest in water samples from tires. Bacterial phylotypes were identified by comparative sequence analysis of 90 16S rDNA DGGE band amplicons. The majority of sequences were placed in five major taxa: Alpha-, Beta- and Gammaproteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, and an unclassified group; Proteobacteria and Bacteroidetes were the predominant heterotrophic bacteria in containers. The bacterial communities in human-made containers consisted mainly of undescribed species, and a phylogenetic analysis based on 16S rRNA sequences suggested that species composition was independent of both container type and the spatial distribution of containers. Comparative PCR-based, cultivation-independent rRNA surveys of microbial communities associated with mosquito habitats can provide significant insight into community organization and dynamics of bacterial species.
Project description:BackgroundCulex pipiens sensu stricto (s.s.) is considered the primary vector of Usutu virus and West Nile virus, and consists of two morphologically identical but behaviourally distinct biotypes (Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus) and their hybrids. Both biotypes are expected to differ in their feeding behaviour, and pipiens/molestus hybrids are presumed to display intermediate feeding behaviour. However, the evidence for distinct feeding patterns is scarce, and to date no studies have related differences in feeding patterns to differences in host abundance.MethodsMosquitoes were collected using CO2-baited traps. We collected blood-engorged Cx. pipiens/torrentium specimens from 12 contrasting urban sites, namely six city parks and six residential areas. Blood engorged Cx. pipiens/torrentium mosquitoes were identified to the species and biotype/hybrid level via real-time polymerase chain reaction (PCR). We performed blood meal analysis via PCR and Sanger sequencing. Additionally, avian host communities were surveyed via vocal sounds and/or visual observation.ResultsWe selected 64 blood-engorged Cx. pipiens/torrentium mosquitoes of which we successfully determined the host origin of 55 specimens. Of these, 38 belonged to biotype pipiens, 14 were pipiens/molestus hybrids and the identity of three specimens could not be determined. No blood-engorged biotype molestus or Cx. torrentium specimens were collected. We observed no differences in feeding patterns between biotype pipiens and pipiens/molestus hybrids across different habitats. Avian community composition differed between city parks and residential areas, whereas overall avian abundance did not differ between the two habitat types.ConclusionsOur results show the following: (1) Cx. pipiens s.s. feeding patterns did not differ between city parks and residential areas, regardless of whether individuals were identified as biotype pipiens or pipiens/molestus hybrids. (2) We detected differences in host availability between city parks and residential areas. (3) We show that in both urban habitat types, biotype pipiens and pipiens/molestus hybrids fed on both mammalian and avian hosts. This underscores the potential role in arbovirus transmission of biotype pipiens and pipiens/molestus hybrids.
Project description:The Asian tiger mosquito Aedes albopictus is well adapted to urban environments and takes advantage of the artificial containers that proliferate in anthropized landscapes. Little is known about the physicochemical, pollutant, and microbiota compositions of Ae. albopictus-colonized aquatic habitats and whether these properties differ with noncolonized habitats. We specifically addressed this question in French community gardens by investigating whether pollution gradients (characterized either by water physicochemical properties combined with pollution variables or by the presence of organic molecules in water) influence water microbial composition and then the presence/absence of Ae. albopictus mosquitoes. Interestingly, we showed that the physicochemical and microbial compositions of noncolonized and colonized waters did not significantly differ, with the exception of N2O and CH4 concentrations, which were higher in noncolonized water samples. Moreover, the microbial composition of larval habitats covaried differentially along the pollution gradients according to colonization status. This study opens new avenues on the impact of pollution on mosquito habitats in urban areas and raises questions on the influence of biotic and abiotic interactions on adult life-history traits and their ability to transmit pathogens to humans.