Taxonomic Evaluation of Leptobrachium hasseltii (TSCHUDI) based on Morphological, Acoustic, and Molecular Analysis

Abstract

Leptobrachium from Southeast Asian populations was first described as Leptobrachium hasseltii Tschudi with type species from Java. Taxonomic reassessment revealed that the Leptobrachium group was a cryptic species due to similarities in their morphological appearance. Leptobrachium hasseltii is then separated into two groups; Sumatran and Javan populations, and Bali populations. Thus, this study aims to evaluate the taxonomic status of L. hasseltii based on morphological characters, bioacoustic and molecular analyses on the populations of Sumatera, Java, and Bali. I analyzed the morphological data, bioacoustic and phylogenetic analyses of L. hasseltii populations from Sumatera, Java, and Bali to evaluate their exact taxonomy position. The genetic material used is mitochondrial DNA gene sequences 16S rRNA. Phylogenetic trees are reconstructed using Maximum Likelihood (ML) and Bayesian Inference (BI) analyses. I used Leptobrachium specimens stored in the Museum Zoologicum Bogoriense (MZB). Approximately 21 morphological characters were measured with digital calipers and an ocular micrometer with a precision of 0.01 mm. Advertisement call characteristics from each population are described, including series of the call, the number of notes per call, note repetition rate, and characteristics of each note. Phylogenetic results shoId Leptobrachium sp. from Bali was separated from all the Sumatran and Javan populations, this result is supported by morphometric data, which its cluster clearly separated from other populations, and its also supported by bioacoustic results, which have a significantly different character. Here I confirmed the possibility of new species Leptobrachium, by combinations of phylogenetic relationship, morphological and morphometry, and bioacoustic analyses.