Abstract
We developed a novel real-time PCR assay for rapid sexing in all three elephant species, which amplifies small fragments of the orthologous sexual chromosome zinc finger protein genes ZFX/ZFY (65 bp). This assay is a simple, inexpensive and reliable tool that is suitable for non-invasive DNA samples and can be incorporated into larger SNP panels for individual identification and population genetic studies.
This is a preview of subscription content, log in via an institution to check access.
Data availability
The datasets generated during during the current study are available from the corresponding author on reasonable request.
References
Ahlering MA, Hailer F, Roberts MT, Foley C (2011) A simple and accurate method to sex savannah, forest and asian elephants using noninvasive sampling techniques. Mol Ecol Resour 11:831–834
Beja-Pereira A, Oliveira R, Alves PC, Schwartz MK, Luikart G (2009) Advancing ecological understandings through technological transformations in noninvasive genetics. Mol Ecol Resour 9:1279–1301
Bourgeois S, Kaden J, Senn H, Bunnefeld N, Jeffery KJ, Akomo-Okoue EF, Ogden R, McEwing R (2019) Improving cost-efficiency of faecal genotyping: new tools for elephant species. PLoS One 14(1):e0210811
Cain B, Watts PC, Wandera AB, Stevens-Wood B, Shawcross SG (2013) A reliable, single-step method for gender determination in black rhinoceros from low-copy template DNA. Conserv Genet Resour 5:655–657
Chakraborty S, Boominathan D, Desai AA, Vidya TNC (2014) Using genetic analysis to estimate population size, sex ratio, and social organization in an Asian elephant population in conflict with humans in Alur, southern India. Conserv Genet 15(4):897–907
Chang H-W, Gu D-L, Su S-H, Chang C-C, Cheng C-A, Huang H-W, Yao C-T, Chou T-C, Chuang L-Y, Cheng C-C (2008) High-throughput gender identification of Accipitridae eagles with real-time PCR using TaqMan probes. Theriogenology 70:83–90
Chelliah K, Bukka H, Sukumar R (2013) Modeling harvest rates and numbers from age and sex ratios: a demonstration for elephant populations. Biol Conserv 165:54–61
Fernando P, Melnick DJ (2001) Molecular sexing eutherian mammals. Mol Ecol Resour 1:350–353
Gupta SK, Thangaraj K, Singh L (2006) A simple and inexpensive molecular method for sexing and identification of the forensic samples of elephant origin. J Forensic Sci 51:805–807
Matejusová I, Bland F, Hall AJ, Harris RN, Snow M, Douglas A, Middlemas SJ (2013) Real-time PCR assays for the identification of harbor and gray seal species and sex: a molecular tool for ecology and management. Mar Mamm Sci 29:186–194
Mondol S, Mailand CR, Wasser SK (2014) Male biased sex ratio of poached elephants is negatively related to poaching intensity over time. Conserv Genet 15:1259–1263
Morin PA, Nestler A, Rubio-Cisneros NT, Robertson KM, Mesnick SL (2005) Interfamilial characterization of a region of the ZFX and ZFY genes facilitates sex determination in cetaceans and other mammals. Mol Ecol 14:3275–3286
Munshi-South J (2011) Relatedness and demography of African forest elephants: inferences from noninvasive fecal DNA analyses. J Hered 102:391–398
Sukumar R, Ramakrishnan U, Santosh JA (1998) Impact of poaching on an Asian elephant population in Periyar, southern India: a model of demography and tusk harvest. In: Animal Conservation Forum. Cambridge University Press, pp. 281–291
Vidya TNC, Kumar VR, Arivazhagan C, Sukumar R (2003) Application of molecular sexing to free-ranging Asian elephant (Elephas maximus) populations in southern India. Curr Sci 85:1074–1077
Acknowledgements
We thank the Centre National de la Recherche Scientifique et Technologique (CENAREST; permit n° AR0016/14/MESRS/CENAREST/CG/CST/CSAR) for research authorization to conduct this study in Gabon. We thank the Agence Nationale des Parcs Nationaux and the Institut de Recherche en Ecologie Tropicale of CENAREST for hosting the study and providing institutional and logistical support. We thank West Midlands Safari Park for providing savannah elephant blood samples collected from routine health monitoring.
Code availability
Not applicable.
Funding
Funds for the project were provided by the Economic Commmunity of Central African States (CEEAC) under the ECOFAC V Programme (Fragile ecosystems of Central Africa) funded by the European Union (contract n° FED/2013/332-349), by the French Development Agency under the France-Gabon debt conversion agreement (Convention AFD CGA 1188.01.H/Projet Eléphant Gabon) and by the Department for Environment, Food and Rural Affaires (DEFRA), UK, as part of their comitment to tackling Illegal Wildlife Trade.
Author information
Authors and Affiliations
Contributions
S.B. and R.M. designed the study. R.M. designed the primers. S.B., J.K., P.K. and T.V. performed the laboratory work. R.M. and K.O. supervised the laboratory work. S.B. performed the statistical analyses. S.B. wrote the manuscript. All co-authors revised the manuscript.
Corresponding author
Ethics declarations
Ethics approval
This research study was conducted in Gabon under permit n° AR0016/14/MESRS/CENAREST/CG/CST/CSAR delivered by the Centre National de la Recherche Scientifique et Technologique (CENAREST). All applicable international, national and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Conflicts of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by: Maciej Szydlowski
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Bourgeois, S., Ouitavon, K., Kongmee, P. et al. A simple sexing test for elephant species and its application to faecal DNA. J Appl Genetics 62, 507–509 (2021). https://doi.org/10.1007/s13353-021-00627-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13353-021-00627-2