Low prevalence of naturalWolbachiain major malarial vectorsAnopheles culicifaciess.l., andAnopheles stephensi:a first report

Sankar, S.; Sundari, T Mowna; Anand, Asha

doi:10.1101/2020.11.22.393652

Cited by 4 publications

(2 citation statements)

References 74 publications

(147 reference statements)

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“…In order to obtain a sufficient number of reads for mapping to the high number of genes across the Wolbachia genome, an optimization in the ddRAD-Seq library preparation (e.g., restriction enzyme selection) should be performed both for the host and the Wolbachia genomes. An alternative to the utilization of 16S rRNA gene marker for Wolbachia detection, a study of Sankar et al (2021) detected low prevalence of natural Wolbachia from Anopheles culicifacies and Anopheles stephensi using a nested PCR method of 16S rRNA. Regarding the evolutionary rates of the 16S rRNA and wsp markers, the detection of Wolbachia using PCR assays showed different results between targets genes where wsp gene were detected from higher number of individuals (83 individuals) than 16S rRNA (49 individuals) (Table 3).…”

Section: Discussionmentioning

confidence: 99%

Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq

Muharromah,

Reyes,

Kagia

et al. 2023

Front. Cell. Infect. Microbiol.

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BackgroundWolbachia, an endosymbiotic bacterium, is globally used to control arboviruses because of its ability to block arboviral replication and manipulate the reproduction of Wolbachia host, Aedes aegypti. Polymerase chain reaction (PCR)-based Wolbachia detection has been recently reported from natural Ae. aegypti populations. However, due to the technical limitations of PCR, such as primer incompatibility, PCR-based assays are not sufficiently reliable or accurate. In this study, we examined double digestion restriction site-associated DNA sequencing (ddRAD-Seq) efficiency and limitations in Wolbachia detection and quantification in field-collected Ae. aegypti natural populations in Metro Manila, the Philippines, compared with PCR-based assays.MethodsA total of 217 individuals Ae. aegypti were collected from Metropolitan Manila, Philippines. We separated it into 14 populations consisting of 7 female and male populations. We constructed a library for pool ddRAD-Seq per population and also screened for Wolbachia by PCR assays using wsp and 16S rRNA. Wolbachia density per population were measured using RPS17 as the housekeeping gene.ResultsFrom 146,239,637 sequence reads obtained, 26,299 and 43,778 reads were mapped across the entire Wolbachia genome (with the wAlbA and wAlbB strains, respectively), suggesting that ddRAD-Seq complements PCR assays and supports more reliable Wolbachia detection from a genome-wide perspective. The number of reads mapped to the Wolbachia genome per population positively correlated with the number of Wolbachia-infected individuals per population based on PCR assays and the relative density of Wolbachia in the Ae. aegypti populations based on qPCR, suggesting ddRAD-Seq-based semi-quantification of Wolbachia by ddRAD-Seq. Male Ae. aegypti exhibited more reads mapped to the Wolbachia genome than females, suggesting higher Wolbachia prevalence rates in their case. We detected 150 single nucleotide polymorphism loci across the Wolbachia genome, allowing for more accurate the detection of four strains: wPip, wRi, TRS of Brugia malayi, and wMel.ConclusionsTaken together, our results demonstrate the feasibility of ddRAD-Seq-based Wolbachia detection from field-collected Ae. aegypti mosquitoes.

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Section: Discussionmentioning

confidence: 99%

Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq

Muharromah,

Reyes,

Kagia

et al. 2023

Front. Cell. Infect. Microbiol.

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“…culicifacies and An. stephensi species from Tamil Nadu [ 14 ]. However, there can be significant overlap in Wolbachia strains infecting one host, and different strains can affect the survival of the other [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Molecular Evidence of Wolbachia Species in Wild-Caught Aedes albopictus and Aedes aegypti Mosquitoes in Four States of Northeast India

Vinayagam,

Nirmolia,

Chetry

et al. 2023

Journal of Tropical Medicine

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Wolbachia, a Gram-negative intracellular bacterium, naturally infects many arthropods, including mosquito vectors responsible for the spread of arboviral diseases such as Zika, chikungunya, and dengue fever. Certain Wolbachia strains are involved in inhibiting arbovirus replication in mosquitoes, and this phenomenon is currently being studied to combat disease vectors. A study was conducted in four states in north-eastern India to investigate the presence of natural Wolbachia infection in wild-caught Aedes albopictus and Aedes aegypti mosquitoes, the established vectors of dengue. The detection of a Wolbachia infection was confirmed by nested PCR and sequencing in the two mosquito species Ae. aegypti and Ae. albopictus. Positivity rates observed in Ae. aegypti and Ae. albopictus pools were 38% (44 of 115) and 85% (41 of 48), respectively, and the difference was significant (chi-square = 28.3174, p = 0.00000010). Sequencing revealed that all detected Wolbachia strains belonged to supergroup B. Although Wolbachia infection in Ae. aegypti has been previously reported from India, no such reports are available from north-eastern India. Data on naturally occurring Wolbachia strains are essential for selecting the optimal strain for the development of Wolbachia-based control measures. This information will be helpful for the future application of Wolbachia-based vector control measures in this part of the country.

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Wolbachia 16S rRNA haplotypes detected in wild Anopheles stephensi in eastern Ethiopia

et al. 2022

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Background About two out of three Ethiopians are at risk of malaria, a disease caused by the parasites Plasmodium falciparum and Plasmodium vivax. Anopheles stephensi, an invasive vector typically found in South Asia and the Middle East, was recently found to be distributed across eastern and central Ethiopia and is capable of transmitting both P. falciparum and P. vivax. The detection of this vector in the Horn of Africa (HOA) coupled with widespread insecticide resistance requires that new methods of vector control be investigated in order to control the spread of malaria. Wolbachia, a naturally occurring endosymbiotic bacterium of mosquitoes, has been identified as a potential vector control tool that can be explored for the control of malaria transmission. Wolbachia could be used to control the mosquito population through suppression or potentially decrease malaria transmission through population replacement. However, the presence of Wolbachia in wild An. stephensi in eastern Ethiopia is unknown. This study aimed to identify the presence and diversity of Wolbachia in An. stephensi across eastern Ethiopia. Methods DNA was extracted from An. stephensi collected from eastern Ethiopia in 2018 and screened for Wolbachia using a 16S targeted PCR assay, as well as multilocus strain typing (MLST) PCR assays. Haplotype and phylogenetic analysis of the sequenced 16S amplicons were conducted to compare with Wolbachia from countries across Africa and Asia. Results Twenty out of the 184 mosquitoes screened were positive for Wolbachia, with multiple haplotypes detected. In addition, phylogenetic analysis revealed two superclades, representing Wolbachia supergroups A and B (bootstrap values of 81 and 72, respectively) with no significant grouping of geographic location or species. A subclade with a bootstrap value of 89 separates the Ethiopian haplotype 2 from other sequences in that superclade. Conclusions These findings provide the first evidence of natural Wolbachia populations in wild An. stephensi in the HOA. They also identify the need for further research to confirm the endosymbiotic relationship between Wolbachia and An. stephensi and to investigate its utility for malaria control in the HOA. Graphical Abstract

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Low prevalence of naturalWolbachiain major malarial vectorsAnopheles culicifaciess.l., andAnopheles stephensi:a first report

Cited by 4 publications

References 74 publications

Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq

Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq

Molecular Evidence of Wolbachia Species in Wild-Caught Aedes albopictus and Aedes aegypti Mosquitoes in Four States of Northeast India

Wolbachia 16S rRNA haplotypes detected in wild Anopheles stephensi in eastern Ethiopia

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