Population cytogenetic and molecular evidence for existence of a new species in Anopheles fluviatilis complex (Diptera: Culicidae)

Cytogenetic, cross-mating and molecular evidence of four cytological races of Anopheles crawfordi (Diptera: Culicidae) in Thailand and Cambodia

“…Our findings are in agreement with the results of cross-matings among karyotypic forms of other anophelines previously reported by several investigators, i.e., Anopheles vagus [34], A. pullus (= Anopheles yatsushiroensis) [35], A. sinensis [36][37][38][39], Anopheles aconitus [25], Anopheles barbirostris A1 and A2 [11,29], Anopheles campestris-like [30], Anopheles peditaeniatus [31,32], and Anopheles paraliae [40]. Until now, numerous studies have used ribosomal and mitochondrial DNA markers for phylogenetic analysis to determine the relationships among sibling species and/or subspecies of Anopheles [11,27,29,30,[41][42][43][44][45][46][47][48]. Recently, Ngo et al [49] reported that Anopheles dangi is deemed to be a synonym of A. crawfordi based on low mean genetic distance (0.006) of COI, COII and Cyt-b genes of mtDNA and the D3 gene of rDNA derived from specimens collected in south-central Vietnam.…”

Modified PCR-based assay for the differentiation of members of Anopheles fluviatilis complex in consequence of the discovery of a new cryptic species (species V)

“…fluviatilis sensu lato (s.l.) populations, one from district Hardwar, Uttarakhand (30.0° N, 78.2° E), where species T, U, and V are prevalent [8] and the other from district Sundergarh, Odisha (22.1240° N, 84.0432° E), where species S is prevalent in sympatric association with T [1,5], were selected for this study. Anopheles fluviatilis s.l.…”

“…Species S is highly anthropophagic and is a highly efficient malaria vector [2,4,5], whereas species T and U are almost zoophilic, and considered as poor vectors [4,6,7]. Subsequently, in the year 2013, a new cryptic species was reported in this complex which has been designated as species V [8]. This new species is characterized by having two fixed paracentric inversions s1 and S present on arms 2 and 3, respectively of polytene chromosome complement, and also by fixed nucleotide differences in the D3 domain of 28S-and ITS2-rDNA [8].…”

“…Subsequently, in the year 2013, a new cryptic species was reported in this complex which has been designated as species V [8]. This new species is characterized by having two fixed paracentric inversions s1 and S present on arms 2 and 3, respectively of polytene chromosome complement, and also by fixed nucleotide differences in the D3 domain of 28S-and ITS2-rDNA [8]. The biology of this new species and its role in malaria transmission is not investigated so far, except that in Hardwar where the new species was identified, species V had 0.04 human blood index while for species T and U collected from the same area it was 0.0 [8].…”

“…Prior to the discovery of new cryptic species (species V) in the Fluviatilis Complex, an allele-specific PCR assay (ASPCR) was developed by Singh et al [9] for the differentiation of the then known three members of the complex, i.e., species S, T, and U, which was based on species-specific differences in nucleotide sequences in D3 domain of 28S ribosomal DNA (rDNA). The discovery of additional cryptic species (species V) in the Fluviatilis Complex [8] has complicated further studies on the bionomics of An. fluviatilis in the absence of a PCR-based assay for the differentiation of all four members of the Fluviatilis Complex.…”