Fig 1.
Sampling sites for mosquito collection.
The wild bird’s migration sites and stopover in Malaysia used as sampling location of mosquitoes in this study [marked with orange pin]. The study location map was prepared using Natural Earth website: https://www.naturalearthdata.com/downloads/.
Table 1.
Sequences of primers used in this study.
Table 2.
Mosquito Distribution According to Location, Genus, Species, Sex and Pool Number.
Fig 2.
Bands of positive mosquito samples from the gel electrophoresis procedure.
The expected target size is 114 bp, targeting capsid and pre-membrane genes of the WNV. The gel was prepared with 1.5% agarose gel electrophoresis (w/v). Key: L: 100 bp size ladder; +ve ctrl: negative control (deionised distilled water); +ve ctrl: positive control (WNV plasmid); S1–S5: pooled samples of female Cx. vishnui mosquitoes.
Fig 3.
Phylogenetic analysis of WNV detected in the mosquito pools in Malaysia with other global WNV.
The analysis includes 52 WNV nucleotide sequences from other countries, 4 WNV nucleotides sequences of wild birds from Malaysia (presented in the blue box) and 35 sequences from this study (presented in the red box). The tree branches are colored according to the proposed WNV lineages; green: Lineage 1; orange: Lineage 2; other lineages (3, 4a, 4c,5 and 6): purple. The tree was constructed using the maximum likelihood method with the Jukes-Cantor model with 1000 bootstrap replicate. Branches are scaled bootstraps.