Genetic management of wild Asian elephants (Elephas maximus) in Thailand
Rutten, Roxan Gerarda Paula Agnes
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Analysis of mitochondrial DNA from dung samples can provide information about the maternal lineages of groups of wild Asian elephants in Thailand. In this Student Research Project (SRP) 98 DNA samples collected from dung in Kaengkrachan National Park (western forest complex of Thailand) and 58 samples collected in Phukieo Wildlife Sanctuary (eastern forest complex of Thailand) were used for mitochondrial DNA analysis. After testing several primers pairs it was decided to use the MDL5 and MDL3 primers that were also used by Fernando et al. (2003(a)), because these primers gave the best sequencing results. A 570 bp fragment of mitochochondrial DNA that includes part of the d-loop region was analyzed using PCR amplification and sequencing. In Kaengkrachan National Park 4 mitochondrial haplotypes were identified: BN/HT01 (n = 16; 16.33%), BH/HT05 (n = 35; 35.71%), BQ/HT07 (n = 39; 39.80%) and haplotype AE_ AH/HT02 (n = 8; 8.16%). In Phukieo Wildlife Sanctuary 4 different haplotypes were found as well: BH/HT05 (n = 8), haplotype AE_AH/HT02 (n = 44) and AB/HT04 (n = 4). One sample from Phukieo Wildlife sanctuary displayed a haplotype that had not been reported previously by Fernando et al. (2003(a)) and Fickel et al. (2007). This could be a new haplotype: New HTA (n = 1). Heteroplasmia was found in one sample from Phukieo Wildlife Sanctuary. Haplotype AE_AH/HT02 and New HTA were both present in this sample. The presence of heteroplasmia indicates that the haplotype New HTA is most likely a recent mutation. It was not possible to do a quantitative analysis of the data collected in Phukieo Wildlife Sanctuary, because no selection of samples was made. All collected samples were analysed, so there is a possibility some individuals were sampled more than once. In the samples taken from Kaengkrachan National Park the assumption was made that each sample was taken from a different individual, based on the method of selecting samples for analysis. The haplotypes that were found in this SRP correlated with haplotypes found by both Fernando et al. (2003(a)) and Fickel et al. (2007), with the exception of the newly found haplotype New HTA. The presence of haplotype BQ/HT07 (which had previously only been identified on the Malaysian peninsula by Fernando et al. (2003(a)) in the western forest complex, but not in the eastern forest complex indicates there may be some geographical influence on the distribution of mitochondrial haplotypes within Thailand. However, the evidence that was found in this project for a geographical influence was inconclusive and further research remains necessary. The information gathered on the d-loop haplotypes was combined with the information that was gathered on cytochrome b haplotypes by Helmie van Lith in her SRP. Combining this information can contribute to understanding the phylogeny of d-loop haplotypes in the Asian elephant. The data gathered in this SRP made a contiburtion to the database of genetic information on the population of wild Asian elephants in Thailand. This data can be used for further genetic research on Asian elephants and may help to design future conservation programs for the species.