Dataset Information

Molecular Identification and Phylogenetic Analysis of Acetylcholinesterase-2 Gene From Culex pipiens And Culex quinquefasciatus (Diptera: Culicine) in Nigeria.

ABSTRACT: West Nile virus (WNV) is maintained in an enzootic cycle between ornithophilic mosquitoes and birds. There is paucity of information on population density of these vectors and the molecular details in Nigeria. We determined relative abundance and employed molecular methods to detect Culex species. Mosquitoes were caught from April 2013 to February 2014. Genomic DNA extraction was carried out and acetylcholinesterase 2 gene amplified by Polymerase Chain Reaction in different Culex species. Culex pipiens quinquefasciatus and Cx. p. pipiens were identified in different locations, while some other Culex species could not be identified. Culex p. quinquefasciatus were sequenced by Sanger's method. Vector density was 100 Culex species per man hour. Identification of Culex pipiens and Cx. quinquefasciatus from different locations in Ibadan, Nigeria reaffirms the presence of vectors and highlights possibility of enzootic transmission of WNV. Surveillance should be sustained in equine population and bird reservoirs.

SUBMITTER: Onoja AB

PROVIDER: S-EPMC7039529 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Molecular Identification and Phylogenetic Analysis of Acetylcholinesterase-2 Gene From Culex pipiens And Culex quinquefasciatus (Diptera: Culicine) in Nigeria.

Onoja A B AB Maina M M MM Opayele A V AV

Nigerian veterinary journal 20181213 4

West Nile virus (WNV) is maintained in an enzootic cycle between ornithophilic mosquitoes and birds. There is paucity of information on population density of these vectors and the molecular details in Nigeria. We determined relative abundance and employed molecular methods to detect Culex species. Mosquitoes were caught from April 2013 to February 2014. Genomic DNA extraction was carried out and acetylcholinesterase 2 gene amplified by Polymerase Chain Reaction in different Culex speci ...[more]

PMID: 32095033

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Project description:BACKGROUND: Culex spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, Culex pipiens pipiens, Culex pipiens quinquefasciatus, and Culex tarsalis are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of Culex pipiens complex mosquitoes and hybrids in Mexico City. Culex pipiens pipiens preferentially feed on avian hosts; Culex pipiens quinquefasciatus have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of Culex pipiens complex mosquitoes and their hybrids. RESULTS: Mosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as Culex, Culiseta or Ochlerotatus by morphological analysis. Within the genus Culex, specimens were further identified as Culex tarsalis or as belonging to the Culex pipiens complex. Members of the Culex pipiens complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the Ace.2 gene. Culex pipiens quinquefasciatus was the most abundant form collected. CONCLUSIONS: Important WNV vectors species, Cx. p. pipiens, Cx. p. quinquefasciatus and Cx. tarsalis, are all present in Mexico City. Hybrids of Cx. p. pipiens and Cx. p. quinquefasciatus were also collected and identified. The presence and abundance of these WNV competent vectors is a cause for concern. Understanding the distribution of these vectors can help improve viral surveillance activities and mosquito control efforts in Mexico City.

Project description:BackgroundThe genomes of three major mosquito vectors of human diseases, Anopheles gambiae, Aedes aegypti, and Culex pipiens quinquefasciatus, have been previously sequenced. C. p. quinquefasciatus has the largest number of predicted protein-coding genes, which partially results from the expansion of three detoxification gene families: cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST), and carboxyl/cholinesterases (CCE). However, unlike An. gambiae and Ae. aegypti, which have large amounts of gene expression data, C. p. quinquefasciatus has limited transcriptomic resources. Knowledge of complete gene expression information is very important for the exploration of the functions of genes involved in specific biological processes. In the present study, the three detoxification gene families of C. p. quinquefasciatus were analyzed for phylogenetic classification and compared with those of three other dipteran insects. Gene expression during various developmental stages and the differential expression responsible for parathion resistance were profiled using the digital gene expression (DGE) technique.ResultsA total of 302 detoxification genes were found in C. p. quinquefasciatus, including 71 CCE, 196 P450, and 35 cytosolic GST genes. Compared with three other dipteran species, gene expansion in Culex mainly occurred in the CCE and P450 families, where the genes of α-esterases, juvenile hormone esterases, and CYP325 of the CYP4 subfamily showed the most pronounced expansion on the genome. For the five DGE libraries, 3.5-3.8 million raw tags were generated and mapped to 13314 reference genes. Among 302 detoxification genes, 225 (75%) were detected for expression in at least one DGE library. One fourth of the CCE and P450 genes were detected uniquely in one stage, indicating potential developmentally regulated expression. A total of 1511 genes showed different expression levels between a parathion-resistant and a susceptible strain. Fifteen detoxification genes, including 2 CCEs, 6 GSTs, and 7 P450s, were expressed at higher levels in the resistant strain.ConclusionsThe results of the present study provide new insights into the functions and evolution of three detoxification gene families in mosquitoes and comprehensive transcriptomic resources for C. p. quinquefasciatus, which will facilitate the elucidation of molecular mechanisms underlying the different biological characteristics of the three major mosquito vectors.

Project description:Culex species are the most widespread mosquito species across the world and are known to be highly opportunistic, feeding on humans and livestock. They are known to acquire the potential to transmit zoonotic diseases, including Rift Valley Fever (RVF). However, despite their public health significance, they remain understudied in North-western Nigeria, compared to Anophelines. This study was therefore aimed at determining the relative abundance and Multiplex polymerase chain reaction (Multiplex PCR) identification of members of the Culex pipiens complex, in Kura Local Government Area (LGA), North-western, Nigeria. Adult mosquitoes were collected using Center for Disease Control (CDC) miniature light traps from August to October 2019. Mosquitoes were identified using morphological identification keys. Members of the Culex pipiens complex were further identified using Multiplex PCR to assess the presence of sibling species. A total of 413 mosquitoes, belonging to 3 genera, Culex, Anopheles and Aedes were collected. Of this figure, 120 Culex spp. females were collected. Homes with livestock had the highest occurrence of mosquitoes, 123 (61.19%) compared to those without livestock, 78 (38.81%). There was no statistical difference among the two (2) categories of homes (P ≥ 0.005). Culicoides spp. were the most common with 130 collected (65.38%). Again, homes with livestock had the highest occurrence, 85 whilst homes without livestock had 45 of the other flies caught. Multiplex- PCR revealed no expected bands for Cx. quinquefasciatus and Cx. pipiens from the DNA obtained from field collected mosquitoes as confirmed by using genomic DNA of an insectary Culex quinquefasciatus as control. Cx. spp. is presently regarded as a biting nuisance having no significant epidemiological importance. Efforts at its control should be intensified before it is too late. This study provides useful information on the occurrence and multiplex PCR of Culex spp in Kura Local Government Area, North-western Nigeria. These results have implications for the control of Culex spp. mosquito populations and the spread of human, livestock and avian diseases.

Project description:The genomes of three major mosquito vectors of human diseases, including Anopheles gambiae, Aedes aegypti, and Culex pipiens quinquefasciatus, have been previously sequenced. C. p. quinquefasciatus has the largest number of predicted protein-coding genes, which partially results from the expansion of three detoxification gene families: cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST), and carboxylcholinesterases (CCE). However, unlike A. gambiae and A. aegypti, which have large amounts of gene expression data, C. p. quinquefasciatus has limited transcriptomic resources. Knowledge of complete gene expression information is very important for the exploration of the functions of genes involved in specific biological processes. In the present study, the three detoxification gene families of C. p. quinquefasciatus were analyzed for phylogenetic classification and compared with those of three other dipteran insects. Gene expression during various developmental stages and the differential expression responsible for parathion resistance were profiled using the digital gene expression (DGE) technique. Results: A total of 291 detoxification genes were found in C. p. quinquefasciatus, including 70 CCE, 186 P450, and 35 GST genes. Compared with three other dipteran species, gene expansion in Culex mainly occurred in the CCE and P450 families, where the genes of α-esterases, juvenile hormone esterases, and CYP325 of the CYP4 subfamily showed the most pronounced expansion on the genome. A total of 13314 genes were expressed in five DGE libraries. Genes with signal transduction and odorant binding functions were prominently expressed during egg development. Genes involved in proteolysis, glycosphingolipid biosynthesis, and purine metabolism were preferentially expressed at the larval stage. Seventy five percent of the detoxification genes were found to be expressed. One fourth of the CCE and P450 genes were expressed at unique stages, indicating their developmentally regulated expression. Fifteen detoxification genes, including 2 CCEs, 6 GSTs, and 7 P450s, were expressed at higher levels in a parathion-resistant strain than in a susceptible strain. Conclusion: The results of the present study provide new insights into the functions and evolution of three detoxification gene families in mosquitoes and comprehensive transcriptomic resources for C. p. quinquefasciatus, which will facilitate the elucidation of molecular mechanisms underlying the different biological characteristics of the three major mosquito vectors. Raw data were deposited in SRA and assigned accession number SRA049959: http://www.ncbi.nlm.nih.gov/sra?term=SRA049959

Dataset Information

Molecular Identification and Phylogenetic Analysis of Acetylcholinesterase-2 Gene From Culex pipiens And Culex quinquefasciatus (Diptera: Culicine) in Nigeria.

Publications

Molecular Identification and Phylogenetic Analysis of Acetylcholinesterase-2 Gene From <i>Culex pipiens</i> And <i>Culex quinquefasciatus</i> (Diptera: Culicine) in Nigeria.

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