Bonobos’ saliva remaining on the pith of terrestrial herbaceous vegetation can serve as non-invasive wild genetic resources
Evaluating the genetic diversity of natural populations of endangered species is important for conservation. Although the genetic analysis of wildlife usually requires collecting DNA non-invasively, the variety of non-invasive DNA sampling methods is limited for each species. We present a method to obtain DNA of an endangered species, the bonobo (Pan paniscus), in which the pith of the terrestrial herbaceous vegetation (THV) that they consumed was newly utilized. We investigated the (1) frequency of encountering remnant saliva on three types of THV pith; (2) concentrations of DNA in the saliva samples by the real-time quantitative PCR; and (3) rates of positive PCR, accurate genotyping, and allelic drop out by analyzing two autosomal microsatellite loci (D7s817 and D9s910). The number of remnant saliva samples was recorded by following the bonobo groups on a daily basis. The frequency of encountering DNA samples was higher in saliva samples than in fecal samples. More than half of the saliva samples remaining on two types of THV pith provided sufficient concentrations of bonobo DNA (> 200 pg/μl). Rates of positive PCR and accurate genotyping were high, and allelic drop out rate was low when the amount of template DNA was above 200 pg per reaction. Our results suggest that the remnants of bonobo saliva on the pith of THV are a potential resource for obtaining DNA, and better than other kinds of samples from the perspective of the abundant sampling opportunities.
KeywordsBonobo Pan paniscus Conservation Non-invasive DNA sampling Saliva Terrestrial herbivorous vegetation
We thank the Research Centre for Ecology and Forestry and the Ministry of Scientific Research of the Democratic Republic of the Congo for permitting our research. We also thank Dr. T. Sakamaki and local assistants at Wamba for help in our fieldwork, and Dr. T. Yumoto for providing valuable comments on terrestrial herbivorous vegetation, and Dr. H. Imai, Dr. G. Hanya, Dr. N. Suzuki-Hashido, Dr. T. Hayakawa, Ms. M. Hakukawa, and Ms. K. Takano for help in our genetic analysis. We also thank the editor and two reviewers for their reviewing processes and helpful comments for our manuscript. This study was financially supported by Japan Society for the Promotion of Science Grant-in-aid for JSPS fellows (17J09827 to S.I.), and the Leading Graduate Program in Primatology and Wildlife Science of Kyoto University.
SI designed this study, collected DNA samples of animals, took photos of THV, conducted DNA experiments, analyzed the genetic data, wrote and revised the manuscript. KT helped DNA sampling of animals. YK and TF supervised SI to design this study. All authors revised the manuscript and gave final approval for publication.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was approved by the Research Centre for Ecology and Forestry and the Ministry of Scientific Research of the Democratic Republic of the Congo, and followed all laws in the Democratic Republic of the Congo. This study was conformed to the Guidelines for Field Research established by the Ethics Committee of the Primate Research Institute, Kyoto University.
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