Wolbachia infection in wild mosquitoes (Diptera: Culicidae): implications for transmission modes and host-endosymbiont associations in Singapore.
BACKGROUND
Wolbachia are intracellular bacterial endosymbionts found in most insect lineages. In mosquitoes, the influence of these endosymbionts on host reproduction and arboviral transmission has spurred numerous studies aimed at using Wolbachia infection as a vector control technique. However, there are several knowledge gaps in the literature and little is known about natural Wolbachia infection across species, their transmission modes, or associations between various Wolbachia lineages and their hosts. This study aims to address these gaps by exploring mosquito-Wolbachia associations and their evolutionary implications.


METHODS
We conducted tissue-specific polymerase chain reaction screening for Wolbachia infection in the leg, gut and reproductive tissues of wild mosquitoes from Singapore using the Wolbachia surface protein gene (wsp) molecular marker. Mosquito-Wolbachia associations were explored using three methods-tanglegram, distance-based, and event-based methods-and by inferred instances of vertical transmission and host shifts.


RESULTS
Adult mosquitoes (271 specimens) representing 14 genera and 40 species were screened for Wolbachia. Overall, 21 species (51.2%) were found positive for Wolbachia, including five in the genus Aedes and five in the genus Culex. To our knowledge, Wolbachia infections have not been previously reported in seven of these 21 species: Aedes nr. fumidus, Aedes annandalei, Uranotaenia obscura, Uranotaenia trilineata, Verrallina butleri, Verrallina sp. and Zeugnomyia gracilis. Wolbachia were predominantly detected in the reproductive tissues, which is an indication of vertical transmission. However, Wolbachia infection rates varied widely within a mosquito host species. There was no clear signal of cophylogeny between the mosquito hosts and the 12 putative Wolbachia strains observed in this study. Host shift events were also observed.


CONCLUSIONS
Our results suggest that the mosquito-Wolbachia relationship is complex and that combinations of transmission modes and multiple evolutionary events likely explain the observed distribution of Wolbachia diversity across mosquito hosts. These findings have implications for a better understanding of the diversity and ecology of Wolbachia and for their utility as biocontrol agents.
['Ding|Huicong|H|', 'Yeo|Huiqing|H|', 'Puniamoorthy|Nalini|N|http://orcid.org/0000-0003-0651-8356']
D000330:Aedes D000711:Anaplasmataceae Infections D000818:Animals D005075:Biological Evolution D003465:Culex D009033:Culicidae D004269:DNA, Bacterial D005260:Female D054884:Host-Pathogen Interactions D008297:Male D000072138:Mosquito Vectors D016133:Polymerase Chain Reaction D017421:Sequence Analysis D012846:Singapore D000079426:Vector Borne Diseases D020577:Wolbachia
Wolbachia Transmission modes Host-endosymbiont association Tissue-specific polymerase chain reaction Wolbachia surface protein gene Reproductive endosymbiont