Kriangwanich Wannapimol, Buddhachat Kittisak, Poommouang Anocha, Chomdej Siriwadee, Thitaram Chatchote, Kaewmong Patcharaporn, Kittiwattanawong Kongkiat, Nganvongpanit Korakot
Department of Veterinary Biosciences and Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
Excellence Center in Veterinary Bioscience, Chiang Mai University, Chiang Mai, Thailand.
PeerJ. 2021 Jun 25;9:e11689. doi: 10.7717/peerj.11689. eCollection 2021.
Currently, species identification of stranded marine mammals mostly relies on morphological features, which has inherent challenges. The use of genetic information for marine mammal species identification remains limited, therefore, new approaches that can contribute to a better monitoring of stranded species are needed. In that context, the ISSR-HRM method we have proposed offers a new approach for marine mammal species identification. Consequently, new approaches need to be developed to identify individuals at the species level. Eight primers of the ISSR markers were chosen for HRM analysis resulting in ranges of accuracy of 56.78-75.50% and 52.14-75.93% in terms of precision, while a degree of sensitivity of more than 80% was recorded when each single primer was used. The ISSR-HRM primer combinations revealed a success rate of 100% in terms of discrimination for all marine mammals included in this study. Furthermore, ISSR-HRM analysis was successfully employed in determining marine mammal discrimination among varying marine mammal species. Thus, ISSR-HRM analysis could serve as an effective alternative tool in the species identification process. This option would offer researchers a heightened level of convenience in terms of its performance and success rate. It would also offer field practice to veterinarians, biologists and other field-related people a greater degree of ease with which they could interpret results when effectively classifying stranded marine mammals. However, further studies with more samples and with a broader geographical scope will be required involving distinct populations to account for the high degree of intraspecific variability in cetaceans and to demonstrate the range of applications of this approach.
目前，搁浅海洋哺乳动物的物种鉴定主要依赖形态特征，这存在一些固有挑战。利用遗传信息进行海洋哺乳动物物种鉴定的应用仍然有限，因此，需要新的方法来更好地监测搁浅物种。在此背景下，我们提出的ISSR-HRM方法为海洋哺乳动物物种鉴定提供了一种新途径。因此，需要开发新方法来在物种水平上识别个体。选择了8个ISSR标记引物用于HRM分析，精度范围为56.78-75.50%和52.14-75.93%，而使用每个单一引物时记录的灵敏度超过80%。ISSR-HRM引物组合对本研究中所有海洋哺乳动物的鉴别成功率为100%。此外，ISSR-HRM分析成功用于确定不同海洋哺乳动物物种之间的鉴别。因此，ISSR-HRM分析可作为物种鉴定过程中的一种有效替代工具。该方法在性能和成功率方面将为研究人员提供更高的便利性。它还将为兽医、生物学家和其他相关领域人员的野外实践提供更大的便利，使他们在有效分类搁浅海洋哺乳动物时能够更轻松地解读结果。然而，需要进一步开展研究，涉及更多样本和更广泛的地理范围，涵盖不同种群，以考虑鲸类动物种内高度的变异性，并证明该方法的应用范围。
