Detection of Wolbachia followed PCR using the wsp-specific primers wsp-81F and wsp-691R44 from DNA extracted from single female mosquitos (twenty by colony) using PureLink Genomic DNA Mini Kit (Invitrogen, Grand Island, New York, USA) according to the manufacturer’s instructions. To verify the quality of the DNA, the amplification of the cytochrome c oxidase (COI) gene was used according to Folmer et al.45.

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