Geometric morphometrics versus DNA barcoding for the identification of malaria vectors Anopheles dirus and An. baimaii in the Thai-Cambodia border

Abstract

Anopheles (Cellia) dirus Peyton & Harrison and Anopheles baimaii Sallum & Peyton are sibling species within the Dirus complex belonging to the Leucosphyrus group, and have been incriminated as primary vectors of malaria in Thailand. In the present study, DNA barcoding and geometric morphometrics were used to distinguish between An. dirus and An. baimaii in the international border areas, Trat Province, eastern Thailand. Our results revealed that DNA barcoding based on the cytochrome c oxidase subunit I gene could not be used to distinguish An. dirus from An. baimaii. The overlapping values between intra- and interspecific genetic divergence indicated no barcoding gap present for An. dirus and An. baimaii (ranging from 0 to 0.99%). However, the results of the geometric morphometric analysis based on the wing shape clearly distinguished An. dirus and An. baimaii, with 92.42% of specimens assigned to the correct species. We concluded that geometric morphometrics is an effective tool for the correct species identification of these two malaria vectors. Our findings could be used to make entomological surveillance information more accurate, leading to further effective mosquito control planning in Thailand and other countries in Southeast Asia.

Evaluation of Modern Techniques for Species Identification of Lutzia Mosquitoes (Diptera: Culicidae) in Thailand: Geometric Morphometrics and DNA Barcoding

“…Recently, DNA barcoding was applied to aid mosquito identification in Thailand and found that although this technique has high efficiency, some mosquitoes are unable to identify the exact species, such as Anopheles dirus and An. baimaii [ 23 ].…”

“…Eighteen landmarks on the wing vein structure of the Lutzia mosquitoes were identified and digitized for the GM analyses ( Figure 3 ). This study’s landmark placements are based on earlier research that successfully identified certain mosquito species [ 23 , 24 , 35 ].…”

“…Previously, this technique was applied to identify cryptic species of the Anopheles dirus and the An. barbirostris complexes in Thailand, and it was found that GM had a high level of discriminating efficiency in many member species [ 23 , 24 ]. In addition, this modern technique was also proven to be effective with several insects of medical and veterinary significance [ 25 , 26 , 27 , 28 , 29 ].…”

Mitochondrial DNA barcoding of mosquito species (Diptera: Culicidae) in Thailand

“…dirus COI exhibit low interspecific differences [ 63 , 64 ]. Chaiphongpachara et al [ 65 ] recently used DNA barcoding to separate An . dirus and An .…”

Species Identification of the Major Japanese Encephalitis Vectors within the Culex vishnui Subgroup (Diptera: Culicidae) in Thailand Using Geometric Morphometrics and DNA Barcoding

“…However, this method is limited by the need for complete morphological specimens and practitioner expertise [ 10 ]. Currently, effective alternative methods for species identification include geometric morphometrics and DNA barcoding; both may support the standard method to accurately identify mosquito samples that are morphologically similar [ 16 , 17 , 18 , 19 , 20 , 21 , 22 ].…”