Identification of Belgian mosquito species (Diptera: Culicidae) by DNA barcoding

Abstract

Since its introduction in 2003, DNA barcoding has proven to be a promising method for the identification of many taxa, including mosquitoes (Diptera: Culicidae). Many mosquito species are potential vectors of pathogens, and correct identification in all life stages is essential for effective mosquito monitoring and control. To use DNA barcoding for species identification, a reliable and comprehensive reference database of verified DNA sequences is required. Hence, DNA sequence diversity of mosquitoes in Belgium was assessed using a 658 bp fragment of the mitochondrial cytochrome oxidase I (COI) gene, and a reference data set was established. Most species appeared as well-supported clusters. Intraspecific Kimura 2-parameter (K2P) distances averaged 0.7%, and the maximum observed K2P distance was 6.2% for Aedes koreicus. A small overlap between intra- and interspecific K2P distances for congeneric sequences was observed. Overall, the identification success using best match and the best close match criteria were high, that is above 98%. No clear genetic division was found between the closely related species Aedes annulipes and Aedes cantans, which can be confused using morphological identification only. The members of the Anopheles maculipennis complex, that is Anopheles maculipennis s.s. and An. messeae, were weakly supported as monophyletic taxa. This study showed that DNA barcoding offers a reliable framework for mosquito species identification in Belgium except for some closely related species.

A distinct group of north European Aedes vexans as determined by mitochondrial and nuclear markers

“…Of the 16 publicly available Ae. vexans sequences from Belgium present in BoLD (Versteirt et al ., ), two fall within the Group 1 COI sequence variant described herein, demonstrating that this variant is present elsewhere in Europe. Other sequences from Russia (Khrabrova et al ., ) and Hungary (Zittra et al ., ) group together with the other Ae.…”

Using Next-Generation Sequencing for DNA Barcoding: Capturing Allelic Variation in ITS2

“…Length was specific to species and caused difficulty in alignment, leading to large differences between some genera and particularly between subfamilies (Figure 5C). Ancient divergences lead to accumulated differences between Anophelinae and Culicinae species, however the degree of distance between these subfamilies using ITS2 is inflated when compared to mitochondrial genes (Cywinska et al 2006; Versteirt et al 2015; Batovska et al 2016). Alignment-free sequence analysis methods could possibly overcome some of the difficulty associated with comparing alleles of different sizes (Little 2011).…”

DNA reference libraries of French Guianese mosquitoes for barcoding and metabarcoding

“…For mosquitoes, the suitability of the COI gene for species identification was first tested on 37 species occurring in Canada [16]. Since then, barcoding has been used for mosquito species in many parts of the world, including India [17], Iran [18], China [19], Argentina [20], Ecuador [21; 22], Pakistan [23], Singapore [24], Belgium [25], Colombia [26] and Brazil [27]. In most cases, these studies show a high correspondence between morphological species delimitation and mtDNA barcode clusters, but others point out the inability of the method to separate some closely related species distinguished by traditional taxonomy [20].…”