An insight into the molecular identity and distribution of Microhyla taraiensis (Anura, Microhylidae) in Khyber Pakhtunkhwa, Pakistan
The taxonomic identity and distribution of Pakistan’s Microhyla in Khyber Pakhtunkhwa has not been confirmed at species level using molecular data; historical records relied on morphology alone and often referred to the now-restricted M. ornata. A combination of field surveys, morphology, bioacoustics, and mitochondrial DNA was utilized to confirm species identity and map the distribution of Microhyla taraiensis in Khyber Pakhtunkhwa (KP), Pakistan. 125 individuals were collected in 13 districts spanning three climatic zones (arid, semi-arid, and hilly/temperate) from lowland plains to lower montane elevations. Adults and tadpoles were examined morphologically; advertisement calls were recorded from one male; and mitochondrial 16S rRNA and COI fragments were sequenced and analyzed with maximum-likelihood phylogenetics. Three COI sequences from Pakistan were identical to one another and matched the M. taraiensis mitogenome from Nepal, and the 16S sequence formed a well-supported clade with Nepalese M. taraiensis, distinct from M. ornata and M. nilphamariensis. Verified records document M. taraiensis at 19 localities in KP, primarily in shallow, human-modified wetlands (paddy fields, irrigation channels, roadside ditches, floodplain ponds), indicating a wider western distribution along the Himalayan foreland than previously documented. These data constitute the first genetically validated records of M. taraiensis in Pakistan, refine regional biogeography at the Indomalayan–Palearctic boundary, and provide a baseline for future genomic, acoustic, and ecological work. Future efforts should pair genome-scale markers with temperature-standardized call libraries, denser spatial sampling, and curated vouchers to assess population structure, refine range limits, and evaluate potential contact with congeners.
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