Molecular Sexing of The Asian Elephants in The Taman Negara National Park
A molecular study was carried out to investigate on the molecular sexing of Asian elephant (Elephas maximus) from selected population in Taman Negara National Park (TNNP). The main objective of this study were i) to evaluate the sex ratio of Asian elephants from faecal samples of Taman Negara National Park by molecular sexing approach using Y-chromosome gene, ii) to infer phylogenetic relationships of individual Asian elephant from study sites using D-loop region sequences and iii) to infer distribution of haplotype in study area of TNNP based on D-loop region sequences. For the molecular sexing, 44 faecal samples on FTA cards and eight positive controls were amplified using Y-chromosome gene (SRY1 and AMELY2) and X-chromosome gene (PLP1). Male samples have three peaks while female only have one peak. For the mitochondrial DNA D-loop region sequences were proceeded to NCBI BLAST, haplotype and phylogenetic analysis. Results from molecular sexing showed that the sex ratio were 24 male and 20 female. For the mitochondrial DNA, only 19 DNA samples successfully amplified using MDL3 and MDL5 primers. Results from the analysis showed that within the 28 sequences, there are nine haplotypes present from 630 bp of sequence. For phylogenetic relationships analysis, Neighbour-Joining (NJ) showed that all the samples were in monophyletic clade with bootstrap 93% support while Maximum Parsimony (MP) showed polyphyletic clade with two differents clade where samples EM572, EM591, EM574 and EM592 were separated from the other samples. From the results obtained, it can be seen that the D-loop region can be used to evaluate the population genetic structure for Asian elephants in National Park. Further research is advised with more additional samples and wider geographical distribution for knowledge of Asian elephant population genetics, molecular ecology and conservation in TNNP.