Vertical transmission of Zika virus in Culex quinquefasciatus Say and Aedes aegypti (L.) mosquitoes

Atchara Phumee,Usavadee Thavara,Rome Buathong,Apiwat Tawatsin,Siwaporn Boonyasuppayakorn,Proawpilart Intayot,Yutthana Joyjinda,Jakkrawarn Chompoosri,Supaporn Wacharapluesadee,Padet Siriyasatien,Rungfar Boonserm

doi:10.1038/s41598-019-41727-8

Abstract

Several mosquito species have been described as vectors for the Zika virus (ZIKV), such as those in the Aedes, Anopheles, Mansonia and Culex genera. Our previous survey studies were found the ZIKV RNA positive in both male, female and larvae of Culex quinquefasciatus Say and Aedes aegypti (L.) mosquitoes collected from active ZIKV infected patients’ homes in Thailand. Therefore, the aims of this study were to investigate whether ZIKV could be vertically transmitted in Cx. quinquefasciatus, Ae. aegypti and Ae. albopictus. Laboratory and field colonies of these mosquito species were maintained and artificially fed with ZIKV in human blood. Fully engorged mosquitoes (F0) were selected and reared for the vertical transmission study. The subsequent mosquito generations were fed with human blood without the virus. ZIKV in the mosquitoes was detected by hemi-nested RT-PCR and sequencing. C6/36 cells were used to isolate ZIKV from samples that tested positive by hemi-nested RT-PCR. Moreover, ZIKV was identified by immunocytochemical staining 7 days after infection in several organs of infected F0 females, including the salivary glands, midguts, yoke granules and facet cells of the eye. The localization of the ZIKV antigen was identified by the presence of the specific antibody in the salivary glands, midguts, yoke granules and facet cells. ZIKV was detected in female and male Cx. quinquefasciatus until the F6 and F2 generations, respectively. The isolated virus showed cytopathic effects in C6/36 cells by 5 days postinfection. The results suggested that the vertical transmission of ZIKV occurs in Cx. quinquefasciatus in the laboratory. However, we were able to detect the presence of ZIKV in Ae. aegypti in only the F1 generation in both male and female mosquitoes, and Ae. albopictus mosquitoes were not able to vertically transmit the virus at all. Data obtained from this study could be valuable for developing a better understanding of the role of Cx. quinquefasciatus as a potential vector for ZIKV transmission in Thailand and may be useful in creating more effective mosquito vector control strategies in the future.

Highlights

Zika virus (ZIKV) is an arbovirus belonging to the Flaviviridae family and the Flavivirus genus, which includes African and Asian lineages[1,2]
Our results demonstrated that vertical transmission of ZIKV occurs in the Cx. quinquefasciatus mosquito
The study was based on a molecular technique for ZIKV RNA detection, the isolation of viruses in cell culture and an ICC assay for the localization of a ZIKV-specific antigen

Summary

Introduction

Zika virus (ZIKV) is an arbovirus belonging to the Flaviviridae family and the Flavivirus genus, which includes African and Asian lineages[1,2]. Previous studies reported that Aedes mosquitoes such as Ae. africanus, Ae. aegypti, Ae. apicocoargenteus, Ae. furcifer, Ae. vittatus and Ae. luteocephalus are the principal vectors of ZIKV5–9. In Thailand a total of 686 of confirmed Zika case had been reported between January and November 201614, by the Ministry of Public Health (MoPH), especially in Phitsanulok and Chanthaburi provinces where Cx. quinquefasciatus were found naturally ZIKV infections. The MoPH reported the first two indigenous cases of ZIKV-related microcephaly in Thailand[15]. In this study, we determined the potential for the Thai Cx. quinquefasciatus mosquito to vertically transmit ZIKV. Information obtained from this study provides fundamental data regarding whether Ae. aegypti, Ae. albopictus and Cx. quinquefasciatus are capable of vertically transmitting ZIKV in the laboratory.

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Conclusion