Fecal DNA analysis for identifying species and sex of sympatric carnivores: A noninvasive method for conservation on the Tsushima islands, Japan
Fecal DNA analysis for identifying species and sex of sympatric carnivores: A noninvasive method for conservation on the Tsushima islands, Japan
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DOI：
10.1093/jhered/esi124
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发表时间：
2005-11-01
影响因子：
3.1
通讯作者：
Tatara, M
中科院分区：
文献类型：
--
作者：
Kurose, N;Masuda, R;Tatara, M
粪便分析是研究哺乳动物食性和物种特性的有用工具。然而，根据体型相似的哺乳动物粪便的形态特征和内容物来鉴定物种通常是困难的。因此，我们开发了非侵入性的DNA分析方法，使用粪便样本的种类和性别的鉴定的四个小同域食肉动物生活在日本对马岛：豹猫（Felis bengalensis），日本貂（Martes melampus），西伯利亚鼬（Mustela sibirica），野猫（Felis卡图斯）。根据以往系统发育研究的DNA序列数据，设计了种特异性引物，对线粒体细胞色素B基因（112-347 bp）进行PCR扩增，以鉴定种属;对SRY基因（135 bp）进行PCR扩增，以鉴定性别。通过对PCR条件的调整，成功地扩增出了特异性DNA片段，并应用于种属鉴定和性别鉴定。从PCR产物中获得的核苷酸序列与预期的食肉动物物种的细胞色素B序列相对应。该协议的开发可能是一个有价值的工具，在管理和保护的四个食肉动物物种发生在对马群岛。
Fecal analysis is a useful tool for the investigation of food habits and species identity in mammals. However, it is generally difficult to identify the species based on the morphological features and contents of feces deposited by mammals of similar body size. Therefore we developed noninvasive DNA analysis methods using fecal samples for identification of the species and sex of four small sympatric carnivores living on the Tsushima Islands of Japan: the leopard cat (Felis bengalensis), Japanese marten (Martes melampus), Siberian weasel (Mustela sibirica), and feral cat (Felis catus). Based on DNA sequence data from previous phylogenetic studies, we designed species-specific primers for polymerase chain reaction (PCR) amplification of the partial mitochondrial cytochrome b gene (112-347 bp) to identify the species and primers for the partial SRY gene (135 bp) to determine the sex. Due to the adjustment of PCR conditions, those specific DNA fragments were successfully amplified and then applied for species and sex identification. Nucleotide sequences obtained from the PCR products corresponded with cytochrome b sequences of the carnivore species expected. The protocol developed could be a valuable tool in the management and conservation of the four carnivore species occurring on the Tsushima Islands.