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研究生:翁瑞鴻
研究生(外文):Ruei-Hong Wong
論文名稱:共域食肉目動物排遺分子鑑種技術建立與應用
論文名稱(外文):Establishment and Application of a Method for Species Identification from Fecal DNA of Sympatric Carnivores
指導教授:吳海音
指導教授(外文):Hai-Yin Wu
學位類別:碩士
校院名稱:國立東華大學
系所名稱:自然資源與環境學系
學門:環境保護學門
學類:環境資源學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:54
中文關鍵詞:排遺食肉目動物物種鑑別多引子巢式PCR
外文關鍵詞:fecal DNAcarnivoraspecies identificationmultiplex-nested PCR
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排遺是野生動物研究者於野外調查時憑藉的訊息之一。然而,共域小型食肉目動物的排遺並不易單從外觀辨識。近年來,非侵入性DNA採樣技術的應用已成為生態學研究、野生動物經營管理及物種保育上的重要工具之一。本研究目的為發展一個準確而簡便的鑑別方法,以提升排遺DNA辨種的效率。本研究自2009年5月至2010年6月於玉山國家公園東部園區採集到487個排遺樣本,利用多引子巢式聚合酶鏈鎖反應法(multiplex-nested PCR),以一組通用引子,搭配五個分別對五種食肉目具專一性的引子,增幅粒線體DNA上12S rRNA基因的不同區段。上述方法成功鑑別出黃喉貂(Martes flavigula chrysospila)、食蟹獴(Herpestes urva formosanus)、黃鼠狼(Mustela sibirica taivana)及白鼻心(Paguma larvata taivana)四種動物。整體PCR鑑別成功率為67%,其中新鮮樣本(N = 223)成功率為73%,舊樣本成功率為62%,高於僅使用通用引子(13%)。在鑑別出物種的327個排遺中,以黃喉貂最多(72%),其次為食蟹獴(20%)、黃鼠狼(8%),而白鼻心排遺僅有1個。比對不同物種的排遺外觀,研究區黃喉貂與黃鼠狼排遺形狀、內含物及出現位置相似,不易區別,而兩者與食蟹獴排遺的外觀有明顯差異。本研究將有助於減少野外調查時排遺辨識的錯誤,並能成為管理者制定保育措施之參考。
Wildlife researchers usually use animals’ feces as indicators of a species’ presence. However, feces of sympatric small carnivores may be of similar appearance, posing difficulty in species identification in the field. Noninvasive genetic techniques have become an important tool in wildlife research in recent years. The purpose of this study is to develop an effective method for species identification from carnivore fecal DNA. I employed multiplex-nested PCR protocol with a pair of universal primers and five species-specific primers to identify the species origins of the 487 carnivore fecal samples collected in the eastern area of the Yushan National Park from May 2009 to June 2010. All primers were designed to amplify specific fragments of mitochondrial DNA 12S rRNA gene, the size of the fragments differed by species. The method successfully identified four species, including yellow-throated marten (Martes flavigula chrysospila), crab-eating mongoose (Herpestes urva formosanus), Siberian weasel (Mustela sibirica taivana) and Formosan gem-faced civet (Paguma larvata taivana). The overall success rate of species identification was 67%. The success rate of fresh samples (73%, N = 223) was higher than that of old samples (62%, N = 264). Among the 327 samples successfully identified, 72% were of yellow-throated marten. It is difficult to differentiate the feces of the two Mustelidae, yellow-throated marten and Siberian weasel, by appearance alone. But they are more readily to distinguish from the feces of crab-eating mongoose. The result provides an effective and accurate species identification method for field survey on small carnivores.
1. 前言 1
2. 材料與方法 7
2.1 研究地 7
2.2 排遺採集 7
2.3 排遺DNA鑑種 8
2.3.1 DNA萃取(DNA extraction) 9
2.3.2 通用引子PCR 10
2.3.3 多引子nested-PCR 10
2.4 資料分析 11
3. 結果 15
3.1 PCR產物鑑種 15
3.2 各物種排遺外觀特徵與座落基質 16
3.3 野外判種與鑑種失敗的排遺 18
4. 討論 19
4.1 排遺分子鑑種方法 19
4.2 鑑種成功率 20
4.3 玉山國家公園東部園區的食肉目動物排遺特徵 22
4.4 未來研究建議 24
5. 結論 27
6. 引用文獻 29
表附錄 35
圖附錄 45
附錄 53
吳海音。2003。玉山國家公園東部園區大型哺乳動物監測計畫。內政部營建署玉山國家公園管理處。
吳海音。2004a。玉山國家公園東部園區中大型哺乳動物監測計畫。內政部營建署玉山國 家公園管理處。
吳海音。2004b。太魯閣國家公園高山地區動物資源基礎調查。內政部營建署太魯閣國家公園管理處。
吳海音。2005。玉山國家公園東部園區臺灣黑熊及偶蹄目動物群聚研究。內政部營建署 玉山國家公園管理處。
吳海音。2006。玉山國家公園東部園區偶蹄目動物監測計畫。內政部營建署玉山國家公 園管理處。
吳海音。2007。玉山國家公園東部園區偶蹄目動物監測計畫(二)。內政部營建署玉山國家公園管理處。
吳海音。2008。玉山國家公園東部園區南安至抱崖哺乳動物監測及與人類活動的關係。內政部營建署玉山國家公園管理處。
吳海音。2009。玉山國家公園東部園區偶蹄目監測及與中小型食肉目動物分布調查。內政部營建署玉山國家公園管理處。
吳海音。2010。玉山國家公園東部園區遊客與野生動物活動監測計畫。內政部營建署玉山國家公園管理處。
呂光洋。1983。太魯閣國家公園動物生態資源調查。內政部營建署太魯閣國家公園管理處。
李嘉鑫。1995。玉山的動物─哺乳類(修訂版)。內政部營建署玉山國家公園管理處。
李嘉鑫(原著)、鄭錫奇(增修)。2009。玉山的動物─哺乳類(再版)。內政部營建署玉山國家公園管理處。
林曜松。2002。生物多樣性公約與生物多樣性保育。國立臺灣大學生物多樣性研究中心。
林曜松。2007。臺灣野生動物保育與研究之發展。林曜松主編。野生動物保育與研究學 術研討會論文集24-27頁。臺北市。
林曜松、李培芬。1982。玉山國家公園動物生態景觀資源調查報告。內政部營建署玉山國家公園管理處。
林良恭、趙榮台、陳一銘、葉雲吟。1998。自然保護區域資源調查監測手冊─哺乳動物。行政院農業委員會。
祁偉廉。1998。臺灣哺乳動物─野外探險實用大圖鑑。大樹文化事業股份有限公司。臺北市。
莊順安。1994。福山森林生態系三種食肉目動物(麝香貓、食蟹獴、鼬獾)的食性研究。國立臺灣大學動物學研究所碩士論文。
楊勝任。2009。玉山國家公園東部園區植群生態調查計畫。內政部營建署玉山國家公園管理處。
裴家騏。1998。利用自動照相設備記錄野生動物活動模式之評估。臺灣林業科學13(4):317-324。
裴家騏、姜博仁。2003。大武山自然保留區和周邊地區雲豹及其他中大型哺乳動物之現況與保育研究(二)。行政院農業委員會林務局。
裴家騏、陳朝圳、吳守從、滕民強。1997。利用自動照相設備與地理資訊系統研究森林野生動物族群之空間分布。中華林學季刊30(3):279-289。
蔡及文。2007。共域黃喉貂與黃鼠狼之食物資源分配。國立臺灣大學森林環境暨資源學 研究所碩士論文。
鄭錫奇、張簡琳玟。1998。臺灣陸域野生哺乳動物之介紹。自然保育季刊23:18-27。
鄭錫奇、張簡琳紋、張仕緯。2000。南投縣哺乳類(再版)。行政院農委會特有生物研究保育中心。
Adams, J. R. and L. P. Waits. 2007. An efficient method for screening faecal DNA genotypes and detecting new individuals and hybrids in the red wolf (Canis rufus) experimental population area. Conservation Genetic 8: 123-131.
Alda, F., J. Inogés, L. Alcaraz, J. Oria, A. Aranda and I. Doadrio. 2008. Looking for the Iberian lynx in central Spain: a needle in a haystack? Animal Conservation 11: 297-305.
Barja, I. 2009. Decision making in plant selection during the faecal-marking behaviour of wild wolves. Animal Behaviour 77: 489-493.
Bellemain, E. and P. Taberlet. 2004. Improved noninvasive genotyping method: application to brown bear (Ursus arctos) faeces. Molecular Ecology Notes 4: 519-522.
Bextine, B., S. J. Tuan, H. Shaikh, M. Blua and T. A. Miller. 2004. Evaluation of methods for extracting Xylella fastidiosa DNA from the glassy-winged sharpshooter. Journal of Economic Entomology 97: 757-763.
Chapin, F. S., III, E. S. Zavaleta, V. T. Eviner, R. L. Naylor, P. M. Vitousek, H. L. Reynolds, D. U. Hooper, S. Lavorel, O. E. Sala, S. E. Hobbie, M. C. Mack and S. Diaz. 2000. Consequences of changing biodiversity. Nature 405: 234-242.
Casper, R. M., S. N. Jarman, B. E. Deagle, N. J. Gales and M. A. Hindell. 2007a. Detecting prey from DNA in predator scats: A comparison with morphological analysis, using Arctocephalus seals fed a known diet. Journal of Experimental Marine Biology and Ecology 347: 144-154.
Casper, R. M., S. N. Jarman, N. J. Gales and M. A. Hindell. 2007b. Combining DNA and morphological analyses of faecal samples improves insight into trophic interactions: a case study using a generalist predator. Marine Biology 152:815-825.
Chame, M. 2003. Terrestrial mammal feces: a morphometric summary and description. Memorias do Instituto Oswaldo Cruz 98: 71-94.
Colli, L., R. Cannas, A. M. Deiana, G. Gandolfi and J. Tagliavini. 2005. Identification of mustelids (Carnivora: Mustelidae) by mitochondrial DNA markers. Mammalian Biology 70: 384-389.
Crooks, K. R. 2002. Relative sensitivities of mammalian carnivores to habitat fragmentation. Conservation Biology 16: 488-502.
Dalén, L., B. Elmhagen and A. Angerbjörn. 2004a. DNA analysis on fox faeces and competition induced niche shifts. Molecular Ecology 13: 2389-2392.
Dalén, L., A. Götherström and A. Angerbjörn. 2004b. Identifying species from pieces of faeces. Conservation Genetic 5: 109-111.
Dallas, J. F., K. E. Coxon, T. Sykes, P. R. F. Chanin, F. Marshall, D. N. Carss, P. J. Bacon, S. B. Piertney and P. A. Racey. 2003. Similar estimates of population genetic composition and sex ratio derived from carcasses and faeces of Eurasian otter Lutra lutra. Molecular Ecology 12:275-282.
Davison, A., J. D.S. Birks, R. C. Brookes, T. C. Braithwaite and J. E. Messenger. 2002. On the origin of faeces: morphological versus molecularmethods for surveying rare carnivores from their scats. Journal of Zoology 257: 141-143.
Dieffenbach, C. W., T. M. Lowe and G. S. Dveksler. 1993. General concepts for PCR primer design. PCR Methods and Applications 3: S30-S37.
Donadio, E. and S. W. Buskirk. 2006. Diet, morphology, and interspecific killing in Carnivora. The American Naturalist 167: 524-536.
Farrell, L. E., J. Roman and M. E. Sunquist. 2000. Dietary separation of sympatric carnivores identified by molecular analysis of scats. Molecular Ecology 9: 1583-1590.
Fernandes, C. A., C. Ginja, I. Pereira, R. Tenreiro, M. W. Bruford and M. S. Reis. 2008. Species-specific mitochondrial DNA markers for identification of non-invasive samples from sympatric carnivores in the Iberian Peninsula. Conservation Genetic 9: 681-690.
Frantz, A. C., M. Schaul, L. C. Pope, F. Fack, L. Schley, C. P. Muller and T. J. Roper. 2004. Estimating population size by genotyping remotely plucked hair: the Eurasian badger. Journal of Applied Ecology 41: 985-995.
Gemmell, N. J. and S. Akiyama. 1996. An efficient method for the extraction of DNA from vertebrate tissues. Trends in Genetics 12: 338-339.
Haag, T., A. S. Santos, C. D. Angelo, A. C. S. Araujo, D. A. Sana, R. G. Morato, F. M. Salzano and E. Eizirik. 2009. Development and testing of an optimized method for DNA-based identification of jaguar (Panthera onca) and puma (Puma concolor) faecal samples for use in ecological and genetic studies. Genetica 136: 505-512.
Harrison, R. L. 2006. A comparison of survey methods for detecting bobcats. Wildlife Society Bulletin 34: 548-552.
Hausknecht, R., R. Gula, B. Pirga and R. Kuehn. 2007. Urine - a source for noninvasive genetic monitoring in wildlife. Molecular Ecology Notes 7: 208-212.
Hung, C. M., S. H. Li and L. L. Lee. 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.
Kocher, T. D., W. K. Thomas, A. Meyer, S. V. Edwards, S. Paabo, F. X. Villablanca and A. C. Wilson. 1989. Dynamics of mitochondrial DNA evolution in aminals: Amplification and sequencing with conserved primers. Proceedings of the National Academy of Sciences USA 86: 6196-6200.
Kohn, M. H. and R. K. Wayne. 1997. Facts from feces revisited. Trends in Evolution and Ecology 12: 223-227.
Livia, L.,V. Francesca, P. Antonella, P. Fausto and R. Bernardino. 2007. A PCR-RFLP method on faecal samples to distinguish Martes martes, Martes foina, Mustela putorius and Vulpes vulpes. Conservation Genetic 8: 757-759.
LoGiudice, K. and R. S. Ostfeld. 2002. Interactions between mammals and trees: predation on mammal-dispersed seeds and the effect of ambient food. Oecologia 130: 420-425.
Long, R. A., P. MacKay, W. J. Zielinski and J. C. Ray. 2008. Noninvasive survey methods for carnivores. Island Press, Washington, D. C., USA.
McCarthy, K. P., T. K. Fuller, M. Ming, T. M. Mccarthy, L. Waits and K. Jumabaev. 2008. Assessing estimators of snow leopard abundance. Journal of Wildlife Management 72: 1826-1833.
McCullough, D. R., K. C. J. Pei and Y. Wang. 2000. Home range, activity patterns, and habitat relations of Reeves’ muntjacs in Taiwan. Journal of Wildlife Management 64: 430-441.
Miller, B., B. Dugelby, D. Foreman, C. M. del Río, R. Noss, M. Phillips, R. Reading, M. E. Soulé, J. Terborgh and L. Willcox. 2001. The importance of large carnivores to healthy ecosystems. Endangered Species Update 18: 202-210.
Milner, J. M., E. B. Nilsen and H. P. Andreassen. 2007. Demographic side effects of selective hunting in ungulates and carnivores. Conservation Biology 21: 36-47.
Murphy, M. A., L. P. Waits and K. C. Kendall. 2003. The influence of diet on faecal DNA amplification and sex identification in brown bears (Ursus arctos). Molecular Ecology 12: 2261-2265.
Murray, D. L., C. A. Kapke, J. F. Evermann and T. K. Fuller. 1999. Infectious disease and the conservation of free-ranging large carnivores. Animal Conservation 2: 241-254.
Nandini, R. and T. Karthik. 2007. Field observations of yellow-throated marten Martes flavigula feeding on flowers in Meghalaya, North-east India. Small Carnivore Conservation 37: 26-27.
Nonaka, N., T. Sano, T. Inoue, M. T. Armua, D. Fukui, K. Katakura and Y. Oku. 2009. Multiplex PCR system for identifying the carnivore origins of faeces for an epidemiological study on Echinococcus multilocularis in Hokkaido, Japan. Parasitology Research 106: 75-83.
Onorato, D., C. White, P. Zager, L. P. Waits. 2006. Detection of predator presence at elk mortality sites using mtDNA analysis of hair and scat samples. Wildlife Society Bulletin 34: 815-820.
Parr, J. W. K. and J. W. Duckworth. 2007. Notes on diet, habituation and sociality of yellow-throated marten Martes flavigula. Small Carnivore Conservation 36: 27-29.
Pilgrim, K. L., K. S. Mckelvey, A. E. Riddle and M. K. Schwartz. 2005. Felid sex identification based on noninvasive genetic samples. Molecular Ecology Notes 5: 60-61.
Prugh, L. R. and C. E. Ritland. 2005. Molecular testing of observer identification of carnivore feces in the field. Wildlife Society Bulletin 33: 189-194.
Rozhnov, V. V., A. L. Morgulis, M. V. Kholodova and I. G. Meshcherskii. 2008. The use of molecular genetic methods for identification of mustelid species by analyzing feces. Russian Journal of Ecology 39: 70-72.
Sanderson, E. W., K. Redford, C.-L. B. Chetkiewicz, R. A. Medellin, A. R. Rabinowitz, J. G. Robinson and A. B. Taber. 2002. Planning to save a species: the jaguar as a model. Conservation Biology 16: 58-72.
Symondson, W. O. C. 2002. Molecular identification of prey in predator diets. Molecular Ecology 11: 627-641.
Taberlet, P., L. P. Waits and G. Luikart. 1999. Noninvasive genetic sampling: look before you leap. Trends in Evolution and Ecology 14: 323-327.
Tamura, K., J. Dudley, M. Nei and S. Kumar. 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24: 1596-1599.
Treves, A. 2009. Hunting for large carnivore conservation. Journal of Applied Ecology 46: 1350-1356.
Treves, A. and K. U. Karanth. 2003. Human-carnivore conflict and perspectives on carnivore management worldwide. Conservation Biology 17: 1491-1499.
Valenzuela-Galván, D., H. T. Ariya and D. W. Macdonald. 2008. Conservation priorities for carnivores considering protected natural areas and human population density. Biodiversity and Conservation 17: 539-558.
Wang, H. Y., M. P. Tsai, M. C. Tu and S. C. Lee. 2000. Universal primers for amplification of the complete mitochondrial 12S rRNA gene in vertebrates. Zoological Studies 39: 61-66.
Weber, W. and A. Rabinowitz. 1996. A global perspective on large carnivore conservation. Conservation Biology 10: 1046-1054.
Wu, H. Y. 1999. Is there current competition between sympatric Siberian weasels (Mustela sibirica) and ferret badgers (Melogale moschata) in a subtropical forest of Taiwan. Zoological Studies 38: 443-451.
Zabala, J. and I. Zuberogoitia. 2003. Badger, Meles meles (Mustelidae, Carnivora), diet assessed through scat-analysis: a comparison and critique of different methods. Folia Zoology 52: 23-30.
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