Detection of diverse Wolbachia 16S rRNA sequences at low titers from malaria vectors in Kayin state, Myanmar

Sawasdichai, Sunisa; Chaumeau, Victor; Dah, Tee; Kulabkeeree, Thithiworada; Kajeechiwa, Ladda; Phanaphadungtham, Monthicha; Trakoolchengkaew, Muesuwa; Kittiphanakun, Praphan; Akararungrot, Yanada; Oo, Kyi; Delmas, Gilles; White, Nicholas J.; Nosten, François

doi:10.12688/wellcomeopenres.15005.4

Cited by 5 publications

(8 citation statements)

References 43 publications

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“…Given that DNA samples were extracted from the head and thorax of female Anopheles, detection failure in nested PCR is possibly due to low-intensity infection or the reproductive organ speci city of Wolbachia. Therefore, assays with high sensitivity, such as quantitative PCR, may aid in the detection of low-intensity Wolbachia infection [30] [31,32]. Moreover, DNA preparation from the whole body of mosquitoes ensures the inclusion of Wolbachia strains that speci cally infect germ cells.…”

Section: Resultsmentioning

confidence: 99%

Molecular Identification of Native Wolbachia Pipientis in Anopheles Minimus in a Low-Malaria Transmission Area of Umphang Valley Along the Thailand-Myanmar Border

Tongkrajang

Ruenchit

Tananchai

et al. 2020

Preprint

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BackgroundWolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia.MethodsFollowing a two-year survey of malaria vectors in Umphang Valley of Tak Province, Thailand, DNA extracts of female An. minimus, An. peditaeniatus, An. maculatus, and An. dirus were subjected to amplification of the conserved region of the 16S rRNA-encoding gene. The DNA sequences of the amplicons were phylogenetically compared with those of known Wolbachia strains.ResultsAmong four Anopheles spp., amplification was detected in only the DNA samples from An. minimus. The DNA sequencing of amplicons revealed 100% similarity to Wolbachia pipientis, confirming the specificity of amplification. The phylogenetic trees indicate a close relationship with Wolbachia strains in subgroup B.ConclusionTo the best of our knowledge, the data presented herein provide the first molecular evidence of a Wolbachia strain in An. minimus, hereinafter named wAnmi, in a low-malaria transmission area in the Umphang Valley of western Thailand. Further biological characterization is required to examine its potential for malaria transmission control in the field.

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Section: Resultsmentioning

confidence: 99%

Molecular Identification of Native Wolbachia Pipientis in Anopheles Minimus in a Low-Malaria Transmission Area of Umphang Valley Along the Thailand-Myanmar Border

Tongkrajang

Ruenchit

Tananchai

et al. 2020

Preprint

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show abstract

“…Given that DNA samples were extracted from the head and thorax of female Anopheles , detection failure in nested PCR is possibly because of low-intensity infection or the reproductive organ specificity of Wolbachia . Therefore, assays with high sensitivity, such as quantitative PCR, may aid in the detection of low-intensity Wolbachia infection [ 39 , 48 , 49 ]. Moreover, DNA preparation from the whole body of mosquitoes ensures the inclusion of Wolbachia strains that specifically infect germ cells.…”

Section: Discussionmentioning

confidence: 99%

“…minimus and An. maculatus collected from different villages [ 39 ]. Thus, the Wolbachia subgroup B in An.…”

Section: Discussionmentioning

confidence: 99%

“…Collectively, identification of the Wolbachia strain in Anopheles spp. requires further confirmation, in which high-sensitivity assays, such as fluorescent in situ hybridization [ 25 ] and quantitative PCR [ 39 ], whole mosquitoes, and more diverse areas will be included.…”

Section: Discussionmentioning

confidence: 99%

“…Level of Wolbachia 16S rRNA- and Plasmodium 18S rRNA-coding DNA sequence was examined in the same DNA sample as the nested PCR using quantitative real-time PCR (qPCR). The use of W-Specf and W16S primers in qPCR had > 100% efficiency in amplification and detected the Wolbachia 16S rRNA-coding region at concentrations lower than combination of W-Specf and W-Specr did [ 26 , 39 ]. Thus, this study deployed the W-Specf and W16S primers.…”

Section: Methodsmentioning

confidence: 99%

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Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border

et al. 2020

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Background Wolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia. Methods Following a 2-year survey of malaria vectors in Umphang Valley of Tak Province, Thailand, DNA extracts of female An. minimus, An. peditaeniatus, and An. maculatus were subjected to amplification of the conserved region of the 16S rRNA-encoding gene. The DNA sequences of the amplicons were phylogenetically compared with those of known Wolbachia strains. Results Among three Anopheles spp., amplification was detected in only the DNA samples from An. minimus. The DNA sequencing of amplicons revealed 100% similarity to Wolbachia pipientis, confirming the specificity of amplification. The Wolbachia-positive An. minimus samples were devoid of Plasmodium 18S rRNA amplification. The phylogenetic trees indicate a close relationship with Wolbachia strains in subgroup B. Conclusion To the best of our knowledge, the data presented herein provide the first molecular evidence of a Wolbachia strain in An. minimus, hereinafter named wAnmi, in a low-malaria transmission area in the Umphang Valley of western Thailand. Further biological characterization is required to examine its potential for malaria transmission control in the field.

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Diversity of Wolbachia infections in Sri Lankan mosquitoes with a new record of Wolbachia Supergroup B infecting Aedes aegypti vector populations

Wijegunawardana,

Gunawardene,

Abeyewickreme

et al. 2024

Sci Rep

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Wolbachia bacteria are common endosymbionts of insects and have recently been applied for controlling arboviral vectors, especially Aedes aegypti mosquito populations. However, several medically important mosquito species in Sri Lanka were present with limited information for the Wolbachia infection status. Therefore, the screening of Wolbachia in indigenous mosquitoes is required prior to a successful application of Wolbachia-based vector control strategy. In this study, screening of 78 mosquito species collected from various parts of the country revealed that 13 species were positive for Wolbachia infection, giving ~ 17% infection frequency of Wolbachia among the Sri Lankan mosquitoes. Twelve Wolbachia-positive mosquito species were selected for downstream Wolbachia strain genotyping using Multi Locus Sequencing Type (MLST), wsp gene, and 16S rRNA gene-based approaches. Results showed that these Wolbachia strains clustered together with the present Wolbachia phylogeny of world mosquito populations with some variations. Almost 90% of the mosquito populations were infected with supergroup B while the remaining were infected with supergroup A. A new record of Wolbachia supergroup B infection in Ae. aegypti, the main vectors of dengue, was highlighted. This finding was further confirmed by real-time qPCR, revealing Wolbachia density variations between Ae. aegypti and Ae. albopictus (p = 0.001), and between males and females (p < 0.05). The evidence of natural Wolbachia infections in Ae. aegypti populations in Sri Lanka is an extremely rare incident that has the potential to be used for arboviral vector control.

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Detection of diverse Wolbachia 16S rRNA sequences at low titers from malaria vectors in Kayin state, Myanmar

Cited by 5 publications

References 43 publications

Molecular Identification of Native Wolbachia Pipientis in Anopheles Minimus in a Low-Malaria Transmission Area of Umphang Valley Along the Thailand-Myanmar Border

Molecular Identification of Native Wolbachia Pipientis in Anopheles Minimus in a Low-Malaria Transmission Area of Umphang Valley Along the Thailand-Myanmar Border

Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border

Diversity of Wolbachia infections in Sri Lankan mosquitoes with a new record of Wolbachia Supergroup B infecting Aedes aegypti vector populations

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