Individual identification and genetic diversity of Eurasian otters based on microsatellite markers

doi:10.16829/j.slxb.150814

Abstract

Abstract: Population size and genetic diversity are important basic information for threatened species conservation. As a Class II protected animal, Eurasian otter (Lutra lutra) was once widely distributed in China, but has experienced severe population decline and range shrinkage over the 20th century. However, only a few studies have been conducted on Eurasian otters in China. Basic research such as population surveys and genetic diversity studies are urgently needed. We collected 270 otter spraints for DNA extraction during 2019-2020 in Yushu City, Qinghai Province and Qingchuan County, Guangyuan City, Sichuan Province. We used 9 microsatellite loci and the SRY gene to genotype individuals and identify sex. Then we estimated the sizes of the two populations based on individual identification and noninvasive capturerecapture method (Capwire). We also assessed their genetic diversity based on the nine microsatellite markers. We successfully genotyped 67 spraints (24. 8%) with 7 - 9 microsatellites and identified 40 otter individuals. Among them, 10 were from Yushu and 30 were from Qingchuan, with a sex ratio of 4F/5M and 15F/14M respectively. Sex identification failed for two individuals. The estimated population of otters within our study area was 13 (95% CI: 7 - 21) in Yushu City and 75 (95% CI: 59-133) in Qingchuan County. The average observed heterozygosity (H_O) and expected heterozygosity (H_E) of the Yushu population were 0. 680 and 0. 611, while those of the Qingchuan population were 0. 664 and 0. 658, revealing a moderate genetic diversity. Otters in Yushu and Qingchuan have been differentiated significantly (F_ST = 0. 238) and no significant inbreeding was found in both populations (F_IS: Yushu -0. 121, Qingchuan -0. 010). As the first study estimating population size and genetic diversity of Eurasian otters based on fecal DNA in Chinese mainland, our work provides important basic information for otter conservation in China.

Key words: Eurasian otter (Lutra lutra), Fecal DNA, Microsatellite, Population size, Genetic diversity

摘要： 种群数量和遗传多样性是保护濒危物种所需的基础信息。欧亚水獭 (Lutra lutra) 属于国家二级保护动物，曾经在我国分布广泛，但在 20 世纪经历了大规模的种群数量下降和分布区缩减。欧亚水獭相关研究十分缺乏，种群数量调查和遗传多样性等基础研究亟待开展。本研究采用非损伤性取样法，于 2019—2020 年在青海省玉树藏族自治州玉树市和四川省广元市青川县的主要河流共采集 270 个欧亚水獭粪便样品并进行 DNA 提取，基于 9 个微卫星位点和 SRY 基因分子标记进行个体识别和性别鉴定，进而使用非损伤性标志重捕法 Capwire 估计两地水獭的种群数量，并基于微卫星分型数据评估两个种群的遗传多样性。最终共有 67 个粪便样品 (24. 8%) 成功分型 7 ~9 个微卫星位点并用于个体识别，共识别出玉树市 10 个个体、青川县 30 个个体，雌雄性比分别为 4:5、15:14，两地各有 1 个个体未成功鉴定性别。估计研究区域内玉树的水獭种群数量为 13 只 (95% 置信区间：7 ~ 21 只)、青川县的水獭种群数量为 75 只 (95% 置信区间：59 ~ 133 只)。玉树、青川种群的平均观测杂合度 H_O分别为 0. 680、0. 664，平均期望杂合度 H_E分别为 0. 611、0. 658，表明两个水獭种群均具有中等的微卫星遗传多样性。玉树、青川种群的平均 F_ST为 0. 238，平均近亲繁殖系数 F_IS分别为-0. 121、-0. 010，表明两个水獭种群之间分化显著，且种群内近交程度低。本研究是我国大陆地区基于粪便 DNA 对欧亚水獭进行种群数量估计和遗传多样性评估，可为我国水獭的保护提供重要的基础信息。

关键词: 欧亚水獭, 粪便 DNA, 微卫星, 种群数量, 遗传多样性

CLC Number:

Q346

ZHENG Kaidan, WANG Qiaoyun, FAN Pengfei, HAN Xuesong, XIAO Mei, SHEN Limin, DONG Zhengyi, ZHANG Lu. Individual identification and genetic diversity of Eurasian otters based on microsatellite markers[J]. ACTA THERIOLOGICA SINICA, 2024, 44(2): 146-158.

郑凯丹, 汪巧云, 范朋飞, 韩雪松, 肖梅, 谌利民, 董正一, 张璐. 基于微卫星的欧亚水獭个体识别和遗传多样性[J]. 兽类学报, 2024, 44(2): 146-158.

References

Balestrieri A, Remonti L, Prigioni C. 2016. Towards Extinction and Back:Decline and Recovery of Otter Populations in Italy. Cham:Springer International Publishing.
Balestrieri A, Gariano P, Grandinetti M, Verduci F, Gianfranceschi L, Gatti E, Mucci N, Mengoni C, Tremolada P. 2022. Faecal DNAbased genetic survey of a relict Eurasian otter (Lutra lutra) popu lation (Sila Massif, S Italy). Conservation Genetics Resources, 14(4):453-461.
Bodkin J L, Ballachey B E, Cronin M A, Scribner K T. 1999. Population demographics and genetic diversity in remnant and translo cated populations of sea otters. Conservation Biology, 13 (6):1378-1385.
Bouchardy C, Lemarchand C, Boulade Y, Gouilloux N, Rosoux R, Berny P. 2011. Natural recolonization of the Eurasian otter (Lutra lutra) in Massif Central (France). IUCN Otter Spec. Group Bull, 28B:26-29.
Broquet T, Ménard N, Petit E. 2007. Noninvasive population genetics:a review of sample source, diet, fragment length and microsat ellite motif effects on amplification success and genotyping error rates. Conservation Genetics, 8 (1):249-260.
Bryant J V, Gottelli D, Zeng X, Hong X, Chan B P L, Fellowes J R, Zhang Y, Luo J, Durrant C, Geissmann T, Chatterjee H J, Turvey S T. 2016. Assessing current genetic status of the Hainan gibbon using historical and demographic baselines:implications for con servation management of species of extreme rarity. Molecular Ecology, 25 (15):3540-3556.
Cheng Z, Tian X, Zhong Z, Li P, Sun D, Bai J, Meng Y, Zhang S, Zhang Y, Wang L, Liu D. 2021. Reintroduction, distribution, population dynamics and conservation of a species formerly ex tinct in the wild:A review of thirty-five years of successful Milu(Elaphurus davidianus) reintroduction in China. Global Ecology and Conservation, 31:e01860.
Cianfrani C, Lay G L, Maiorano L, Satizábal H F, Loy A, Guisan A. 2011. Adapting global conservation strategies to climate change at the European scale:The otter as a flagship species. Biological Conservation, 144 (8):2068-2080.
Cianfrani C, Maiorano L, Loy A, Kranz A, Lehmann A, Maggini R, Guisan A. 2013. There and back again? Combining habitat suit ability modelling and connectivity analyses to assess a potential return of the otter to Switzerland. Animal Conservation, 16 (5):584-594.
Dallas J, Piertney S. 1998. Microsatellite primers for the Eurasian ot ter. Molecular Ecology, 7:1248-1251.
Dallas J F. 2000. Sex identification of the Eurasian otter Lutra lutra by PCR typing of spraints. Conservation Genetics, 1 (2):181-183.
Dallas J F, Marshall F, Piertney S B, Bacon P J, Racey P A. 2002. Spatially restricted gene flow and reduced microsatellite polymor phism in the Eurasian otter Lutra lutra in Britain. Conservation Genetics, 3 (1):15-28.
Dallas J, Coxon K, Sykes T, Chanin P, Marshall F, Carss D, Bacon P, Piertney S, Racey P. 2003. Similar estimates of population ge netic composition and sex ratio derived from carcasses and faeces of Eurasian otter Lutra lutra. Molecular Ecology, 12:275-282.
Dou H, Yang H, Feng L, Mou P, Wang T, Ge J. 2016. Estimating the population size and genetic diversity of Amur tigers in northeast China. PLoS ONE, 11 (4):e0154254.
Durbin L S. 1998. Habitat selection by five otters Lutra lutra in rivers of northern Scotland. Journal of Zoology, 245 (1):85-92.
Excoffier L, Lischer H E L. 2010. Arlequin suite ver 3. 5:a new series of programs to perform population genetics analyses under Linux and Windows. Molecular Ecology Resources, 10 (3):564-567.
Foster-Turley P, Macdonald S M, Mason C. 1990. Otters:an action plan for their conservation. Gland, Switzerland:IUCN/SSC Otter Specialist Group.
Frankham R, Ballou J D, Briscoe D A. 2002. Introduction to Conser vation Genetics. Cambridge:Cambridge University Press.
Gagneux P, Boesch C, Woodruff D S. 1997. Microsatellite scoring errors associated with noninvasive genotyping based on nuclear DNA amplified from shed hair. Molecular Ecology, 6 (9):861-868.
Galpern P, Manseau M, Hettinga P, Smith K, Wilson P. 2012. Allele match:an R package for identifying unique multilocus genotypes where genotyping error and missing data may be present. Molecular Ecology Resources, 12 (4):771-778.
Han X S, Dong Z Y, Zhao G, Zhao X, Shi X Y, Lü Z, Li H Q. 2021.Using surveillance cameras to analyze the activity pattern of the Eurasian otters (Lutra lutra) and the efficiency of camera trap monitoring. Biodiversity Science, 29 (6):770-779. (in Chinese)HÁjkovÁ P, ZemanovÁ B, Bryja J, HÁjek B, Roche K, Tkadlec E, Zima J. 2006. Factors affecting success of PCR amplification of microsatellite loci from otter faeces. Molecular Ecology Notes, 6(2):559-562.
Hájková P, Zemanová B, Roche K, Hájek B. 2009. An evaluation of field and noninvasive genetic methods for estimating Eurasian ot ter population size. Conservation Genetics, 10 (6):1667-1681.
Honnen A C, Roos A, Stjernberg T, Zachos F E. 2015. Genetic analy sis of Eurasian otters (Lutra lutra) reveals high admixture in Fin land and pronounced differentiation in Sweden. Mammalian Biol ogy, 80 (1):47-53.
Huang C C. 2005. Fecal DNA typing to determine the fine scale population structure and sex-biased dispersal pattern of Eurasian otter(Lutra lutra) in Kinmen. Master thesis. Taipei:Taiwan Univer sity. (in Chinese)
Huang C C, Hsu Y C, Lee L L, Li S H. 2005. Isolation and character ization of tetramicrosatellite DNA markers in the Eurasian otter(Lutra lutra). Molecular Ecology Notes, 5:314-316.
Huang X, Hu N Q, He K, Guan Z H, Garber P A, Chapman C A, Jiang X L, Fan P F. 2022. Disassociation of social and sexual partner relationships in a gibbon population with stable one-male twofemale groups. American Journal of Primatology, 84 (7):e23394.
Hung C M, Li S H, Lee L L. 2004. Faecal DNA typing to determine the abundance and spatial organisation of otters (Lutra lutra)along two stream systems in Kinmen. Animal Conservation, 7:301-311.
Jang-Liaw N H. 2021. A barcodingbased scatanalysis assessment of Eurasian otter Lutra lutra diet on Kinmen Island. Ecology and Evolution, 11 (13):8795-8813.
Jang-Liaw N H, Tan Y C, Chang C J, Juan C H, Hou H Y, Chung L W, Cao H S, Waku D, Chang S W, Lee L L. 2023. Genetic diversity and structure of Eurasian otters on Kinmen Island. Conservation Genetics, 24:589-606.
Janssens X, Fontaine M C, Michaux J R, Libois R, De Kermabon J, Defourny P, Baret P V. 2008. Genetic pattern of the recent recov ery of European otters in southern France. Ecography, 31 (2):176-186.
Kalz B, Jewgenow K, Fickel J. 2006. Structure of an otter (Lutra lutra) population in Germany-results of DNA and hormone analy ses from faecal samples. Mammalian Biology, 71 (6):321-335.
Kruuk H, Conroy J W H, Glimmerveen U, Ouwerkerk E J. 1986. The use of spraints to survey populations of otters Lutra lutra. Bio logical Conservation, 35 (2):187-194.
Kruuk H. 2006. Otters:Ecology, Behaviour and Conservation. Ox ford:Oxford University Press.
Lampa S, Gruber B, Henle K, Hoehn M. 2007. An optimisation ap proach to increase DNA amplification success of otter faeces.Conservation Genetics, 9 (1):201-210.
Lampa S, Mihoub J B, Gruber B, Klenke R, Henle K. 2015. Noninvasive genetic mark-recapture as a means to study population sizes and marking behaviour of the elusive Eurasian otter (Lutra lutra). PLoS ONE, 10 (5):e0125684.
Lanszki J, Hidas A, Szentes K, Révay T, Lehoczky I, Weiss S. 2008.Relative spraint density and genetic structure of otter (Lutra lutra) along the Drava River in Hungary. Mammalian Biology, 73 (1):40-47.
Lanszki J, Hidas A, Szentes K, Révay T, Lehoczky I, Jeney Z, Weiss S. 2010. Genetic structure of otter (Lutra lutra) populations from two fishpond systems in Hungary. Mammalian Biology, 75:447-450.
Lerone L, Mengoni C, Carpaneto G M, Randi E, Loy A. 2014. Proce dures to genotype problematic non-invasive otter (Lutra lutra) samples. Acta Theriologica, 59 (4):511-520.
Lerone L, Mengoni C, Di Febbraro M, Krupa H, Loy A. 2022. A non invasive genetic insight into the spatial and social organization of an endangered population of the Eurasian otter (Lutra lutra, Mustelidae, Carnivora). Sustainability, 14 (4):1943.
Martin E A, Heurich M, Müller J, Bufka L, Bubliy O, Fickel J. 2017.Genetic variability and size estimates of the Eurasian otter (Lutra lutra) population in the Bohemian Forest Ecosystem. Mammalian Biology, 86:42-47.
McMillan S, Wong A, Tang S, Yau E, Gomersall T, Wong P, Vu A, Sin S, Hau B, Bonebrake T. 2022. Spraints demonstrate small population size and reliance on fishponds for Eurasian otter (Lutra lutra) in Hong Kong. Conservation Science and Practice, 5 (1):e12851. DOI:10. 1111/csp2. 12851.
Miller C R, Joyce P, Waits L P. 2005. A new method for estimating the size of small populations from genetic mark-recapture data.
Molecular Ecology, 14 (7):1991-2005.
Mucci N, Arrendal J, Ansorge H, Bailey M, Bodner M, Delibes M, Fer rando A, Fournier P, Fournier C, Godoy J A, Hajkova P, Hauer S, Heggberget T M, Heidecke D, Kirjavainen H, Krueger H-H, Kvaloy K, Lafontaine L, Lanszki J, Lemarchand C, Liukko U-M, Loeschcke V, Ludwig G, Madsen A B, Mercier L, Ozolins J, Pau novic M, Pertoldi C, Piriz A, Prigioni C, Santos-Reis M, Luis T S, Stjernberg T, Schmid H, Suchentrunk F, Teubner J, Tornberg R, Zinke O, Randi E. 2010. Genetic diversity and landscape genetic structure of otter (Lutra lutra) populations in Europe. Conservation Genetics, 11 (2):583-599.
Park C S, Cho G J. 2017. Individual identification of Eurasian otters(Lutra lutra) in South Korea (Sincheon River, Daegu) by micro satellite markers. Journal of Veterinary Medical Science, 79 (6):1064-1067.
Peakall R, Smouse P E. 2012. GenAlEx 6. 5:genetic analysis in Excel. Population genetic software for teaching and research-an up date. Bioinformatics (Oxford, England), 28 (19):2537-2539.
Pennell M W, Stansbury C R, Waits L P, Miller C R. 2013. Capwire:a R package for estimating population census size from noninvasive genetic sampling. Molecular Ecology Resources, 13 (1):154-157.
Piao Z J, Sui Y C, Wang Q, Li Z, Niu L J. 2011. Population fluctua tion and resource protection of otter (Lutra lutra) in Changbai Mountain Nature Reserve. Journal of Hydroecology, 32 (2):115-120. (in Chinese)
Pigneur L M, Caublot G, Fournier-Chambrillon C, Fournier P, Giralda Carrera G, Grémillet X, Le Roux B, Marc D, Simonnet F, Smitz N, Sourp E, Steinmetz J, Urra-Maya F, Michaux J R. 2019. Cur rent genetic admixture between relictual populations might en hance the recovery of an elusive carnivore. Conservation Genetics, 20 (5):1133-1148.
Quaglietta L, Fonseca da Silva V, Hajkova P, Mira A, Boitani L. 2013. Fine-scale population genetic structure and short-range sex-biased dispersal in a solitary carnivore, Lutra lutra. Journal of Mammalogy, 94:561-571.
Quaglietta L, Hájková P, Mira A, Boitani L. 2015. Eurasian otter (Lu tralutra) density estimate based on radio tracking and other data sources. Mammal Research, 60 (2):127-137.
R Core Team. 2023. R:A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria.
Raesly E. 2001. Progress and status of river otter reintroduction projects in the United States. Wildlife Society Bulletin, 29:856-862.
Ran J H, Liu S Y, Sun Z Y, Liu S C, Wang H J, Liu X G, He W H. 2002. Main associated animals in giant panda habitat in Qingchuan County, Sichuan. Sichuan Journal of Zoology, 21 (1):50-52. (in Chinese)
Räikkönen J, Bignert A, Mortensen P, Fernholm B. 2006. Congenital defects in a highly inbred wild wolf population (Canis lupus).Mammalian Biology, 71 (2):65-73.
Ruiz-Olmo J, Jimenez J, López-Martín J. 1995. Radio-tracking of otters Lutra lutra in north-eastern Spain. Lutra, 38:11-21.
Ruiz-Olmo J, Saavedra D, Jiménez J. 2001. Testing the surveys and visual and track censuses of Eurasian otters (Lutra lutra). Journal of Zoology, 253 (3):359-369.
Sittenthaler M, Schöll E M, Leeb C, Haring E, Parz-Gollner R, Hackländer K. 2020. Marking behaviour and census of Eurasian otters (Lutra lutra) in riverine habitats:what can scat abundances and non-invasive genetic sampling tell us about otter numbers?Mammal Research, 65 (2):191-202.
Sulkava R T, Sulkava P O, Sulkava P E. 2007. Source and sink dynamics of density-dependent otter (Lutra lutra) populations in riv ers of central Finland. Oecologia, 153 (3):579-588.
Taberlet P, Griffin S, Goossens B, Questiau S, Manceau V, Escaravage N, Waits L, Bouvet J. 1996. Reliable genotyping of samples with very low DNA quantities using PCR. Nucleic Acids Research, 24:3189-3194.
Tison JL, Blennow V, Palkopoulou E, Gustafsson P, Roos A, Dalén L. 2015. Population structure and recent temporal changes in ge netic variation in Eurasian otters from Sweden. Conservation Genetics, 16 (2):371-384.
Wang Q Y, Zheng K D, Han X S, He F, Zhao X, Fan P F, Zhang L. 2021. Site-specific and seasonal variation in habitat use of Eur asian otters (Lutra lutra) in western China:implications for con servation. Zoological Research, 42 (6):825-832.
Weinberger I C, Muff S, de Jongh A, Kranz A, Bontadina F. 2016.Flexible habitat selection paves the way for a recovery of otter populations in the European Alps. Biological Conservation, 199:88-95.
Wilson D E, Reeder D M. 2005. Mammal Species of the World:A Taxonomic and Geographic Reference. 3rd ed. Baltimore, Mary land:Johns Hopkins University Press.
Zhang B, Li M, Zhang Z, Goossens B, Zhu L, Zhang S, Hu J, Bruford M W, Wei F. 2007. Genetic viability and population history of the giant panda, putting an end to the "evolutionary dead end"?Molecular Biology and Evolution, 24 (8):1801-1810.
Zhang L, Fan P F. 2020. Conservation status of otters in China and a discussion on restoring otter populations in the Pearl River Delta.Acta Theriologica Sinica, 40 (1):71-80. (in Chinese)
Zhang L, Wang Q, Yang L, Li F, Chan B P L, Xiao Z, Li S, Song D, Piao Z, Fan P. 2018. The neglected otters in China:Distribution change in the past 400 years and current conservation status. Biological Conservation, 228:259-267.
Zheng K D. 2022. Individual identification and genetic diversity of Eurasian otter based on molecular markers. Master thesis.Guangzhou:Sun Yat-sen University. (in Chinese)
冉江洪, 刘少英, 孙治宇, 刘世昌, 王鸿加, 刘小庚, 何万红. 2002. 四川青川县大熊猫栖息地主要伴生哺乳动物调查. 四川动物, 21(1):50-52.
朴正吉, 睢亚橙, 王群, 李卓, 牛丽君. 2011. 长白山自然保护区水獭种群数量变动与资源保护. 水生态学杂志, 32 (2):115-120.
张璐, 范朋飞. 2020. 中国水獭保护现状及珠江口水獭种群重建探讨. 兽类学报, 40 (1):71-80.
郑凯丹. 2022. 基于分子标记的欧亚水獭个体识别和遗传多样性研究. 广州:中山大学硕士学位论文.
黄传景. 2005. 利用排遗 DNA 标定法探讨金门地区水獭之族群遗传结构与雌雄播迁模式之差异. 台北:台湾大学硕士论文.
韩雪松, 董正一, 赵格, 赵翔, 史湘莹, 吕植, 李宏奇. 2021. 基于视频监控系统的欧亚水獭活动节律初报及红外相机监测效果评估. 生物多样性, 29 (6):770-779.

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