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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林恩仲(Ec-Chung Lin) | |
dc.contributor.author | Yue-Jheng Lyu | en |
dc.contributor.author | 呂岳錚 | zh_TW |
dc.date.accessioned | 2021-06-15T01:21:06Z | - |
dc.date.available | 2010-07-30 | |
dc.date.copyright | 2009-07-30 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-24 | |
dc.identifier.citation | 李俊億、謝幸媚。2008。親子鑑定的演算邏輯,第 26 頁。臺大出版中心,台北市。
胡忠一。2005。日本實施食品產銷履歷制度與我國推動示範該畫辦理現況。農政與農情 154: 47-52。 胡忠一。2006。我國農產品產銷履歷制度推動現況。農業貿易人才訓練班 出口實務班講義。Available: http://coa.cpc.org.tw/edu/Class/doc/95/%E8%AC%9B% E7%BE%A9/95%E5%88%9D%E9%9A%8E2/%E8%BE%B2%E7%94%A2%E5%93%81%E7%94%A2%E9%8A%B7%E5%B1%A5%E6%AD%B7%E6%8E%A8%E5%8B%95%E7%8F%BE%E6%B3%81_%E8%83%A1%E5%BF%A0%E4%B8%80%E7%A7%91%E9%95%B7_.pdf. Access: Dec. 2008. 胡忠一。2008。農產品產銷履歷制度。綠色農業科技研討會講義。Available: http://www.nttu.edu.tw/apic/20080505/paper/2.doc. Access: Dec. 2008. 徐立衡。2008。USDA 表示試驗計畫顯示動物辨識有用。台灣農產品安全資訊追溯網。Available: http://taft.coa.gov.tw/public/data/8121615572271.pdf. Access: Dec. 2008. 張靜文。2005。加拿大牛隻生產履歷之介紹。農政與農情 160: 88-93。 陳雅琴。2005。美國農業部籌建全國動物標識系統。行政院農業委員會。Available: http://www.coa.gov.tw/htmlareafile/web-articles/4881/0403.pdf. Access: Dec. 2008. 陳雅琴。2006。美國食品安全的制度體系與法令規定。農政與農情 164: 77-82。 陳祈睿。2008。我國推動農產品產銷履歷之執行成果。農政與農情 188: 44-47。 黃培訓。2004。日本食品生產履歷對臺灣的啟示。農政與農情 145: 75-82。 鄭祝菁。2008。豬隻產銷履歷推動現況與展望。農政與農情 192: 44-46。 龔榮太。2007。世界各國農畜產品積極導入產銷履歷之探討。畜產報導月刊 88: 42-50。 Alves, E., C. Castellanos, C. Ovilo, L. Silio, and C. Rodrı’guez. 2002. Differentiation of the raw material of the Iberian pig meat industry based on the use of amplified fragment length polymorphism. Meat Sci. 61: 157–162. Arana, A., B. Soret, I. Lasa, and L. Alfonso. 2002. Meat traceability using DNA markers: Application to the beef industry. Meat Sci. 61: 367–373. Bai, L., C. Ma, S. Gong, and Y. Yang. 2007. Food safety assurance systems in China. Food Control 18: 480-484. Ballester, M., A. Mercade, B. van Haandel, J. Santamartina, and A. Sanchez. 2007. Individual identification and genetic traceability in pig using the SMPlexTM genotyping system. Anim. Genet. 38: 662-666. Barcos, L. O. 2001. Recent developments in animal identification and the traceability of animal products in international trade. Revue scientifique et technique 20: 640-651. Bernard, D.1998. Developing and implementing HACCP in the USA. Food Control 9: 91-95. Bjørnstad, G., and K. H. Røed. 2002. Evaluation of factors affecting individual assignment precision using microsatellite data from horse breeds and simulated breed crosses. Anim. Genet. 33: 264-270. Blasi, M., A. Lanza, C. Varlotta, A. Rosati, and S. Dall’Olio. 2003. Pig ham genetic traceability. Ital. J. Anim. Sci. 2(Suppl. 1): 82-84. Brookes, A. J. 1999. The essence of SNPs. Gene 234: 177-186. Bostein, D., R. L. White, M. Skolnick, and R.W. Davis. 1980. Construction of a genetic linkage map using restriction fragment length polymorphisms. Am. J. Hum. Genet. 32: 314-331. Cornuet, J. M., S. Piry, G. Luikart, A. Estoup, and M. Solignac. 1999. New methods employing multilocus genotypes to select or exclude populations as origins of individuals. Genetics 153: 1989-2000. Cunningham, E. P., and C. M. Meghen. 2001. Biological identification systems: genetic markers. Rev. Sci. Tech. 20: 491-499. Dalvit, C., M. De Marchi, and M. Cassandro. 2007. Genetic traceability of livestock products: A review. Meat Sci. 77: 437-449. Dalvit, C., M. De. Marchi, C. Targhetta, M. Gervaso, and M. Cassandro. 2008. Genetic traceability of meat using microsatellite markers. Food Res. Int. 41: 301-307. Eichmann, C., B. Berger, M. Steinlechner, and W. Parson. 2005. Estimating the probability of identity in a random dog population using 15 highly polymorphic canine STR markers. Forensic Sci. Int. 151: 37-44. European Commission. 1996. Commission Regulation (EEC) No. 1081/96 of 14 June 1996. Reference Method for the detection of cows’ milk and caseinate in cheeses from ewes’ milk, goats’ milk and buffalos’ milk. Official Journal of the European Communities, L142, 15–25. European Commission. 1997. Green Paper on the general principles of food law in the European Union. European Commission. 2000. White Paper on food safety. European Commission. 2000. Council Regulation (EC) No. 1760/2000 of 17 July 2000 establishing a system for the identification and registration of bovine animals and regarding the labeling of beef and beef products and repealing Council Regulation (EC) No 820/97. OfficialJournal of the European Communities, L 204, 1–10. European Commission. 2000. Commission Regulation (EC) No. 1825/2000 of 25 August 2000 laying down detailed rules for the application of Regulation (EC) No 1760/2000 of the European Parliament and of the Council as regards the labelling of beef and beef products. Official Journal of the European Communities, L 216, 8–12. European Commission. 2002. Council Regulation (EC) No 178/2002 of 28 January 2002 laying down the general principles and requirementsof food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety. Official Journal of the European Communities, L 31, 1-24. Fahrenkrug, S. C., B. A. Freking, T. P. L. Smith, G. A. Rohrer, and J. W. Keele. 2002. Single nucleotide polymorphism (SNP) discovery in porcine expressed genes. Anim. Genet. 33: 186-195. Fan, B., Y. Z. Chen, C. Moran, S. H. Zhao, B. Liu, M. Yu, M. J. Zhu, T. A. Xiong, and K. Li. 2005. Individual-breed assignment analysis in Swine populations by using microsatellite markers. Asian-Aust. J. Anim. Sci. 18: 1529-1534. Fan, B., Z. G. Wang, Y. J. Li, X. L. Zhao, B. Liu, S. H. Zhao, M. Yu, M. H. Li , S. L. Chen, T. A. Xiong, and K. Li. 2002. Genetic variation analysis within and among Chinese indigenous swine populations using microsatellite markers. Anim. Genet. 33: 422-427. Fang, M., X. Hu, T. Jiang, M. Braunschweig, L. Hu, Z. Du, J. Feng, Q. Zhang, C. Wu, and N. Li. 2005. The phylogeny of Chinese indigenous pig breeds inferred from microsatellite markers. Anim. Genet. 36: 7-13. Fung, W. K., and Y. Q. Hu. 2008. Statistical DNA forensic: Theory, Methods and Computation. Page 25-27. John Wiley and Sons Press. New York. Geldermann, H., E. Müller, G. Moser1, G. Reiner, H. Bartenschlager, S. Cepica, A. Stratil, J. Kuryl, C. Moran, R. Davoli, and C. Brunsch. 2003. Genome-wide linkage and QTL mapping in porcine F2 families generated from Pietrain, Meishan and wild boar crosses. J. Anim. Breed Genet. 120: 363-393. Gemmell, N. J., and S. Akiyama. 1996. An efficient method for the extraction of DNA from vertebrate tissues. Trends Genet. 12: 338-339. Goffaux, F., B. China, L. Dams, A. Clinquart, and G. Daube. 2005. Development of a genetic traceability test in pig based on single nucleotide polymorphism detection. Forensic Sci. Int. 151: 239-247. Hafler D. A., and P. L. De Jager. 2005. Applying a new generation of genetic maps to understand human inflammatory disease. Nat. Rev. Immunol. 5: 83-91. Harris, H. 1966. Enzyme polymorphism in man. Proc. Roy. Sac.164: 298-310. Heaton, M. P., G. P. Harhay, G. L. Bennett, R. T. Stone, W. M. Grosse, and E. Casas. 2002. Selection and use of SNP markers for animal identification and paternity analysis in U.S. beef cattle. Mammal. Genome 13: 272-281. Heaton, M. P., J. E. Keen, M. L. Clawson, G. P. Harhay, N. Bauer, and C. Schultz. 2005. Use of bovine single nucleotide polymorphism markers to verify sample tracking in beef processing. J. Am. Vet. Med. Assoc. 226: 1311-1314. Horng, Y. M., and M. C. Huang. 2003. Male-specific DNA sequences in pigs. Theriogenology 59: 841-848 Hunter, R. L., and C. L. Markert. 1957. Histonchemical demonstration of enzymes separated by zone electrophoresis in starch gels. Science 125: 1294-1295. International Organization for Standardization (ISO). 1994. ISO 8402: Quality management and quality assurance – vocabulary. Geneva: ISO. Jacquiet, P., J. F. Humbert, A. M. Comes, J. Cabaret, A. Thiam, and D. Cheikh. 1995. Ecological, morphological and genetic characterization of sympatric Haemonchus spp. Parasites of domestic ruminants in Mauritania. Parasitology 110: 483-492. Jeffreys, A. J., J. F. Y. Brookfield, and R. Semeonoff. 1985a. Positive identification of an immigration test case using DNA fingerprints. Nature 317: 818-819. Jeffreys, A. J., V. Wilson, and S. L. Thein. 1985b. Hypervariable 'minisatellite' regions in human DNA. Nature 314: 76-79. Jeffreys, A. J., V. Wilson, and S. L. Thein. 1985c. Individual-specific 'fingerprints' of human DNA. Nature 316: 76-79 Kacirek, S. L., K. M. Irvin, P. I. Dimsoski, S. J. Moeller, M. E. Davis, and H. C. Hines. 1998. Variation at microsatellite loci in the Large White, Yorkshire and Hampshire breeds of swine. Proceedings of the 6th World Congress on Genetics Application to Livestock Production 23: 640-643. Kan, Y. W., and A. M. Dozy. 1978. Polymorphism of DNA sequence adjacent to human beta-globin structural gene: relationship to sickle mutation. Proc. Natl. Acad. Sci. U.S.A. 75: 5631-5635. Katsuaki, S., and O. Takashi. 2008. Cattle traceability system in Japan for BSE. Vet. Ital. 44: 519-526. Kimura, M., and J. F. Crow. 1964. The number of alleles that can be maintained in a finite population. Genetics 49: 725-738. Kim, S., and A. Misra. 2007. SNP genotyping: technologies and biomedical application. Annu. Rev. Biomed. Eng. 9: 289-320. Kim, T. H., K. S. Kim, B. H. Choi, D. H. Yoon, G.. W. Jang, K. T. Lee, H. Y. Chung, H. Y. Lee, H. S. Park, and J. W. Lee. 2005. Genetic structure of pig breeds from Korea and China using microsatellite loci analysis. J. Anim. Sci. 83: 2255-2263. Kirkpatrick, B. W. 1992. Identification of a conserved microsatellite site in the porcine and bovine insulin-like growth factor-I gene 5' flank. Anim. Genet. 23: 543-548. Latorra D. and M. S. Schanfield. 1996. Analysis of human specificity in AFLP systems APOB, PAH, and D1S80. Forensic Sci. Int. 83: 15-25. Li, C. C. 1955. Population Genetics. Page 297-298. University of Chicago Press. Chicago. Martinez, I., and I. Malmheden Yman. 1998. Species identification in meat products by RAPD analysis. Food Res. Int. 31: 459–466. Michelmore, R. W., I. Paran, and R. V. Kesseli. 1991. Identification of markers linked to disease-resistance genes by bulked segregate analysis: a rapid method to detect markers in specific regions by using segregating populations. Proc. Natl. Acad. Sci. U.S.A 88: 9828-9832. Miraglia M., K. G. Berdalb, C. Breraa, P. Corbisierc, A. Holst-Jensenb, E. J. Kokd, H. J. P. Marvind, H. Schimmelc, J. Rentsche, J. P. P. F. van Ried, and J. Zagonf. 2004. Detection and traceability of genetically modified organisms in the food production chain. Food and Chem. Toxic. 42: 1157-1180. Mullis M. B., and F. A. Faloonas. 1987. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 155: 335-350. Nandi, S., P. K. Subudhi, and D. Senadhira. 1997. Mapping QTLs for submerqence tolerance in rice by AFLP analysis and selective genotyping. Mol. Gen. Genet. 255: 1-8. Nei, M. 1978. Estimation of average heterozygosity and genetic distance from a small number of individual. Genetic 89: 583-590. Nei, M. 1987. Molecular evolutionary genetics. Page 87-88. Columbia University Press. New York. Nechtelberger, D., C. Kaltwasser, I. Stur, J. -N. Meyer, G. Brem, M. Mueller, and S. Mueller. 2001. DNA microsatellite analysis for parentage control in Austrian pigs. Anim. Biotech. 12: 141-144. Orrú, L., F. Napolitano, G. Catillo, and B. Moioli. 2006. Meat molecular traceability: How to choose the best set of microsatellites? Meat Sci. 72: 312-317. Rohrer, G. A., L. J. Alexander, J. W. Keele, T. P. Smith, and C. W. Beattie. 1994. A microsatellite linkage map of the porcine genome. Genetics 136: 231-245. Rohrer, G. A., B. A. Freking, and D. Nonneman. 2007. Single nucleotide polymor- phisms for pig identification and parentage exclusion. Anim. Genet. 38: 253-258. Rajic, A., L. A. Waddell, J. M. Sargeant, S. Read, J. Farber, M. J. Firth, and A. Chambers. 2007. An overview of microbial food safety programs in beef, pork, and poultry from farm to processing in Canada. J. Food Prot. 70: 1286-1294. Saez, R., Y. Sanz, and F. Toldra. 2004. PCR-based fingerprinting techniques for rapid detection of animal species in meat products. Meat Sci. 66: 659-665. Saiki, R. K., S. Scharf, F. A. Faloona, K. B. Mullis, G. T. Horn, H. A. Erlich, and N. Arnheim. 1985. Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350-1354 SanCristobal, M., C. Chevalet, C. S. Haley, R. Joosten, A. P. Rattink, B. Harlizius, M. A. M. Groenen, Y. Amigues, M.-Y. Boscher, G. Russell, A. Law, R. Davoli, V. Russo, C. De´saute´s, L. Alderson, E. Fimland, M. Bagga, J. V. Delgado, J. L. Vega-Pla, A. M. Martinez, M. Ramos, P. Glodek, J. N. Meyer, G. C. Gandini, D. Matassino, G. S. Plastow, K. W. Siggens, G. Laval, A. L. Archibald, D. Milan, K. Hammond, and R. Cardellino. 2006a. Genetic diversity within and between European pig breeds using microsatellite markers. Anim. Genet. 37: 189-198. SanCristobal, M., C. Chevalet, J. Peleman, H. Heuven, B. Brugmans, M. van Schriek, R. Joosten, R. A. P. Rattink, B. Harlizius, M. A. M. Groenen, Y. Amigues, M. Y. Boscher, G. Russell, A. Law, R. Davoli, V. Russo, C. Desautes, L. Alderson, E. Fimland, M. Bagga, J. V. Delgado, J. L. Vega-Pla, A. M. Martinez, M. Ramos, P. Glodek, J. N. Meyer, G. Gandini, G. D. Matassino, K. Siggens, G. Laval, A. Archibald, D. Milan, K. Hammond, R. Cardellino, C. Haley, and G. Plastow. 2006b. Genetic diversity in European pigs utilizing amplified fragment length polymorphism markers. Anim. Genet. 37: 232-238. Schwagele, F. 2005. Traceability from a European perspective. Meat Sci. 71: 164-173. Schembri, N., J. A. Toribio, F. Sithole, and P. K. Holyoake. 2007. Review of identification and traceability legislation for pigs in Australia. Aust. Vet. J. 85: 255-260 Shackelford, E. S. D., J. W. Keele, M. Koohmaraie, T. P. Smith, R. T. Stone. and J. V. Delgado. 2003. Detection of quantitative trait loci for growth and carcass composition in cattle carcass. J. Anim. Sci. 81: 2976-2983. Shackell, G. H., H. C. Mathias, V. M. Cave, and K. G. Dodds. 2005. Evaluation of microsatellites as a potential tool for product tracing of ground beef mixtures. Meat Sci. 70: 337–345. Smithies, O. 1955. Zone electrophoresisin starch gels: Group variation in the serum proteins of normal individuals. Biochem. J. 61: 629-641. Smith, G. C., D. L. Pendell, J. D. Tatum, K. E. Belk, and J. N. Sofos. 2008. Post-slaughter traceability. Meat Sci. 80: 66-74. Sobrio, B., M. Brio´n, and A. Carracedo. 2005. SNPs in forensic genetics: a review on SNP typing methodologies. Forensic Sci. Int. 154: 181-194. Southern, E. M. 1970. Base sequence and evolution of guinea-pig alpha-satelllite DNA. Nature 227: 794-798. Tautz, D. 1989. Hypervariability of simple sequences as a general source for polymorphic DNA markers. Nucleic Acids Res. 17: 6463-6471. Ueno, S., N. Tomaru, H. Yoshimaru, T. Manabe, and S. Yamamoto. 2002. Size-class differences in genetic structure and individual distribution of Camellia japonica L. in a Japanese old-growth evergreen forest. Heredity 89: 120-126. Umar, A., J. C. Boyer, D. C. Thomas, D. C. Nguyen, J. I. Risinger, J. Boyd, Y. Ionov, M. Perucho, and T. A. Kunkel. 1994. Defective mismatch repair in extracts of colorectal and endometrial cancer cell lines exhibiting microsatellite instability. J. Biol. Chem. 269: 14367-14370. Vicente, A. A., M. I. Carolino, M. C. O. Sousa, C. Ginja, F. S. Silva, A. M. Martinez, L. Vega-Pla, N. Carolino, and L. T. Gama. 2008. Genetic diversity in native and commercial breeds of pigs in Portugal assessed by microsatellites. J. Anim. Sci. 86: 2496-2507. Vos, P., and R. B. Hogers. 1995. AFLP: A new technique for DNA fingerprinting. Nueleic Acids Res. 23: 4407-4414. Waits, P. L., L. Gordon, and T. Pierre. 2001. Estimating the probability of identity among genotypes in natural populations: cautions and guidelines. Mol. Ecol. 10: 249-256. Wang. D. G. 1998. Large-scale identification, mapping, and genotyping of single-nucleotide polymorphisms in the human.genome. Science 280: 1077-82. Weissenbach, J., G. Gyapay, C. Dib, A. Vignal, J. Morissette, P. Millasseau, G. Vaysseix, and M. J. Lathrop. 1994. A second-generation linkage map of the human genome. Biol. Chem. 269: 14367-14370. Williams J. G. K., A. R. Kubelik, J. Livak, A. Rafalski, and S. V. Tingey. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Res. 18: 6531-6535. Wright, S. 1978. Evolution and the Genetics of Populations. vol. 4. Variability Within and Among Natural Populations. Page 53-56. University of Chicago Press. Chicago. Wyman, A. R., and R. A. White. 1980. A highly polymorphic locus in human DNA. Proc. Natl. Acad. Sci. U.S.A. 77: 6754-6758. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42728 | - |
dc.description.abstract | 近年來由於狂牛病的爆發及各種食品安全事件,使得食品安全日漸受到重視,故最近世界各國以歐盟為首,都在推行產銷履歷制度以增進食品安全。傳統使用在動物製品上的產銷履歷以紙本記錄及耳標為主,但是此兩種方法均有造假或錯置的可能,因此利用 DNA 分子標幟進行遺傳追溯對傳統產銷履歷方法進行驗證成為近年來的研究方向之一,由於微衛星標幟具有高多態性、廣泛分布於真核生物染色體及易於使用聚合酶鏈鎖反應 (polymerase chain reaction, PCR) 技術複製等優點,使之常被用來做為遺傳追溯的分子標幟,本研究之目的即在建立可用於台灣地區豬隻產銷履歷驗證之微衛星標幟組。
本研究對來自檢定站,台灣地區常見的三個品種的純種豬隻 (藍瑞斯, 約克夏及杜洛克) 以及來自數個私人豬場之商用雜交肉豬 (藍瑞斯×杜洛克 (LD),藍瑞斯×約克夏×杜洛克 (LYD) 及未知遺傳背景雜交肉豬 (UnP) 採用10 組微衛星標幟進行研究,計算各標幟之個體鑑別率 (probability of identity, P(ID)) 以確定其個體鑑別之效率,實驗方法為對微衛星標幟引子標記螢光進行兩組多引子PCR 增殖,並進行毛細管電泳以進行基因型鑑定,結果發現三個純品種其理論異質度 (expected heterozyosity, HE) 在無遺傳相關個體上分別為 0.62、0.65 及 0.65;在所有個體間 (部分具有遺傳相關) 分別為 0.61、0.66 及 0.65。其多態性訊息含量 (polymorphism information content, PIC) 在無遺傳相關個體上分別為0.57、0.60 及 0.60;在所有個體間 (部分具有遺傳相關) 分別為 0.57、0.61 及 0.61,三個純品種均在 S0227 有最低的遺傳岐異而在 SW857 有最高的遺傳岐異,進行哈溫平衡 (Hardy-Weinberg equilibrium, HWE) 檢定的結果,在無遺傳相關個體上,藍瑞斯及約克夏各有一個標幟,杜洛克有三個標幟偏離 HWE (P<0.05),在所有個體間 (部分具有遺傳相關),藍瑞斯有一個標幟,約克夏及杜洛克各有三個標幟偏離 HWE (P<0.05),但無論有無遺傳相關,十個標幟之總合均無族群偏離 HWE,表示少數標幟偏離 HWE 對個體鑑別之影響並不顯著。 進行個體鑑定之結果,三個品種其 P(ID) 在遺傳相關個體上分別為 1.42×10-8、6.49×10-9 及 1.23×10-8,在所有個體間 (部分具有遺傳相關) 分別為 1.58×10-8、5.61×10-9 及 9.34×10-9 ,而對雜交豬隻進行個體鑑別之結果,LD 雜交肉豬、LYD 雜交肉豬及 UnP 雜交肉豬其 P(ID) 分別為1.79×10-9、6.14×10-9 及 3.83×10-9,所有族群之 P(ID) 值均小於1×10-7。 另於屠宰場採集民間兩豬場 14 批未知品種肉豬,共 339 頭,與大賣場取得之零售樣品共 8 批進行產銷履歷驗證,可以發現大賣場採得之同批樣品內均有相同之基因型,但僅有一批大賣場樣品可追溯到正確批次之屠宰場採集樣品。其餘之大賣場採樣樣品均未能對應到正確之屠宰場採樣樣品,這些樣品有些具有銷售日期與屠宰日期相差過遠之現象,或是經基因型鑑定的結果,出現了屠宰場採樣樣品所來自之兩豬場所無或極少出現之交替基因。 綜上所述,臺灣近五年之每年上市屠宰的純種淘汰猪及雜交肉猪總數在一千萬頭以下,故屠宰猪在無論有無遺傳相關的情況下,此 10 組微衛星標幟應可達成臺灣常見之純種猪及雜交肉猪的個體鑑別需求,以應用於未來新鮮猪肉產品的產銷履歷回溯驗證。 | zh_TW |
dc.description.abstract | In the last few decades, Bovine spongiform encephalopathy (BSE) and other food safety issues induced to increase consumer’s concerns of safety in food supply chain. EU member countries and other nations in the world have regulations to ensure food traceability in order to promote food safety. Conventional traceability of animal products consists of labeling system and paper documents, which could be counterfeit and less reliable. To verify the correctness of conventional traceability, genetic traceability based on DNA markers has become popular in recently year. Because microsatellite markers have advantage such as highly polymorphic, abundantly dispersed throughout most eukaryotic nuclear genome, and efficiently amplified using polymerase chain reaction (PCR), they are often utilized as molecular markers in genetic traceability. The objective of this study is to establish the microsatellite marker sets for genetic verification traceability of fresh pork products in Taiwan.
In this study, purebred pigs of three breeds (Landrace, Yorkshire, and Duroc) from the central test station and commercial crossbred pigs (Landrace × Duroc (LD), Landrace × Yorkshire × Duroc (LYD), and unknown genetic background crossbred pigs (UnP) from several private farms were genotyped using ten microsatellite marker sets. The probability of identity (P(ID)) of each marker set in individual genetic traceability was calculated. We performed two multiplex polymerase chain reaction (PCR) with fluorescent-labeled microsatellite markers. The PCR products were then separated by capillary electrophoresis and genotyping. The expected heterozygosity (HE) in none genetic related individuals of three pure breeds were 0.62, 0.65, and 0.65; in all individuals (partial genetic related) of three pure breeds were 0.61, 0.66, and 0.65. The polymorphism information content (PIC) in unrelated individuals of the three breeds were 0.57, 0.60, and 0.60; in all individuals (partial genetic related) of three breeds were 0.57, 0.61, and 0.61. In the three breeds, the genetic diversity was the lowest in the marker of S0227 and the highest in SW857. After performing Hardy-Weinberg equilibrium (HWE) test in the three breeds, in unrelated individuals of the three breeds, there were one in Landrace, one in Yorkshire, and three in Duroc microsatellite markers against HWE respectively (P<0.05), in all individuals (partial genetic related) of three breeds, there were one in Landrace, three in Yorkshire and three in Duroc microsatellite markers against HWE respectively (P<0.05). The combination of observed heterozygosity (HO) with ten microsatellite marker sets showed that the overall situation still agreed with HWE. Therefore, those marker sets against HWE had no significant effects on individual identification. The results of individual identification showed that P(ID) in unrelated individuals of three breeds were 1.42×10-8, 6.49×10-9, and 1.23×10-8; in all individuals (partial genetic related) of three breeds were 1.58×10-8, 5.61×10-9, and 9.34×10-9. The P(ID) of LD crossbred pigs, LYD crossbred pigs, and UnP were 1.79×10-9, 6.14×10-9, and 3.83×10-9, respectively. All the tested groups had their P(ID) values less than 1.00×10-7. We collected 339 UnP with 14 batches from two different private farms in slaughter house, and 8 batches of retail sale sample from hypermarkets for traceability test. The result showed that samples from hypermarkets with the same batch number had the same genotypes. Two batches from hypermarkets could trace back to batches from slaughter house correctly. Other samples from hypermarket could not correspond to correct sample collected in slaughter house, some of the samples had issues of the sales date far from slaughtering date, the results of genotyping, the alleles appeared in these samples were rare or even never appeared in these two farms. In Taiwan, the number of purebred and crossbred pigs slaughtered per year has been less than ten million (1×107) heads in recent five years. Thus, according the results of this study these ten microsatellite markers are sufficient for individual identification among the common purebred and crossbred commercial pigs in the genetic traceability of fresh pork. | en |
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dc.description.tableofcontents | 誌謝....................................................................................................................................Ι
目錄..................................................................................................................................II 圖次..................................................................................................................................V 表次.................................................................................................................................VI 中文摘要......................................................................................................................VIII 英文摘要..........................................................................................................................X 壹、前言...........................................................................................................................1 貳、文獻檢討.....................................................................................................................2 一、產銷履歷 (Traceability) 的重要性.......................................................................2 二、產銷履歷之原則及精神.........................................................................................3 三、各國產銷履歷之執行現況.....................................................................................4 (一) 歐洲地區..........................................................................................................4 (二) 北美地區..........................................................................................................5 1. 美國..................................................................................................................5 2. 加拿大..............................................................................................................8 (三) 亞洲地區..........................................................................................................8 1. 日本..................................................................................................................8 2. 台灣..................................................................................................................9 (四) 其他地區........................................................................................................14 四、分子遺傳標記.......................................................................................................14 (一) 等位基因酶 (Allozyme)................................................................................15 (二) DNA 分子遺傳標記.......................................................................................15 1. 限制片段長度多態性 (Restriction Fragment Length Polymorphism, RFLP). .............................................................................................................................16 2. 隨機增殖 DNA 多態性 (Random Amplified Polymorphism DNA, RAPD). .............................................................................................................................16 3. 增殖片段長度多態性 (Amplified Fragment Length Polymorphism, AFLP).. .............................................................................................................................17 4. 微衛星 (Microsatellite).................................................................................18 5. 單核苷酸多態性 (Single Nucleotide Polymorphism, SNP).........................19 (三) 族群遺傳統計..............................................................................................20 1. 有效交替基因數 (Number of effective alleles, Ne)......................................20 2. 理論異質度 (Expected heterozygosity, HE)..................................................21 3. 觀測異質度 (Observed heterozygosity, HO)............................................... 21 4. 多態性訊息含量 (Polymorphism Information Content, PIC)......................22 5. 懷特氏固定指數 (Wright’s fixation index)..................................................22 五、分子遺傳標記於產銷履歷上之運用...................................................................23 六、研究動機...............................................................................................................26 參、材料與方法...............................................................................................................28 一、試驗樣本..............................................................................................................28 二、基因組 DNA 萃取...............................................................................................28 三、微衛星標幟引子設計...........................................................................................30 四、聚合酶鏈鎖反應 (Polymerase Chain Reaction, PCR).........................................31 五、電泳樣品製備.......................................................................................................33 六、PCR產物基因型判別...........................................................................................33 七、微衛星標幟基因型頻率統計...............................................................................33 肆、結果...........................................................................................................................35 一、PCR產物增殖結果...............................................................................................35 二、純種豬隻之遺傳多態性 .....................................................................................35 三、純種豬隻之 HWE 檢定及 FIS 值......................................................................44 四、純種豬隻之個體鑑定...........................................................................................47 五、雜交豬隻之個體鑑別...........................................................................................50 六、產銷履歷之驗證...................................................................................................53 伍、討論...........................................................................................................................56 一、純種豬隻微衛星標幟多態性之討論...................................................................56 二、純種豬隻之 HWE 檢定......................................................................................57 三、純種豬隻之個體鑑別...........................................................................................58 四、雜交豬隻之個體鑑別 .........................................................................................60 五、驗證結果之探討...................................................................................................61 六、未來研究方向.............................................................................................62 陸、結論...........................................................................................................................63 柒、參考文獻...................................................................................................................64 圖次 頁次 圖 1. 產銷履歷流程圖....................................................................................................4 圖 2. 產銷履歷農產品 TAP, OTAP及UTAP 標章 ..................................................10 圖 3. 各微衛星標幟其PCR產物所標定之螢光及其片段大小..................................34 圖 4. 毛細管電泳訊號經 Peak Scanner 1.0 分析之波型圖.......................................39圖 5. 純品種豬隻 P(ID) 對標幟數之折線圖...............................................................52 圖 6. 純品種及雜交豬隻 P(ID) 對標幟數之折線圖....................................................52 表次 頁次 表 1. 歐盟近年推行之食品安全措施一覽表................................................................7 表 2. 美國近年推行之食品安全措施一覽表................................................................7 表 3. 日本近年推行之產銷履歷措施一覽表................................................................9 表 4. 本實驗所使用豬隻品種及 DNA 樣品來源......................................................29 表 5. 屠宰場採樣之日期、批號及樣品數....................................................................30 表 6. 大賣場採樣之地點、批號、產品類型及採樣號..................................................31 表 7. 本試驗 PCR 反應所使用之微衛星標幟、螢光種類及其引子序列.................32 表 8. 本試驗所觀測到各微衛星標幟之交替基因 (PCR 產物片段大小)................38 表 9. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體在 10 組微衛星標幟之Na.....................................................................................................................40 表 10. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之全部之個體在 10 組微衛星標幟之Na...............................................................................................................40 表 11. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體在 10 組微衛星標幟之 Ne.............................................................................................................41 表 12. 藍瑞斯、約克夏及杜洛克三個品種豬隻之全部之個體在 10 組微衛星標幟之 Ne.................................................................................................................41 表 13. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體在 10 組微衛星標幟之 HE 及其標準機差 (SE).........................................................................42 表 14. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之全部之個體在 10 組微衛星標幟之 HE 及其標準機差 (SE)..........................................................................42 表 15. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體在 10 組微衛星標幟之 PIC...........................................................................................................43 表 16. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之全部之個體在 10 組微衛星標幟之 PIC...........................................................................................................43 表 17. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體 10 個微衛星標幟之 HO................................................................................................................45 表 18. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之全部之個體 10 個微衛星標幟之 HO................................................................................................................45 表 19. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體 10 個微衛星標幟之 FIS.................................................................................................................46 表 20. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之全部之個體 10 個微衛星標幟之 FIS.................................................................................................................46 表 21. 藍瑞斯、約克夏及杜洛克三個純品種豬隻之無相關個體 10 個微衛星標幟之 P(ID)..............................................................................................................48 表 22. 藍瑞斯、約克夏及杜洛克三個純品種豬隻全部之個體 10 個微衛星標幟之 P(ID)....................................................................................................................48 表 23. 藍瑞斯、約克夏及杜洛克三個純品種豬隻無相關個體 10 個微衛星標幟之 P(ID)sibs................................................................................................................49 表 24. 藍瑞斯、約克夏及杜洛克三個純品種豬隻全部之個體 10 個微衛星標幟之 P(ID)sibs................................................................................................................49 表 25. 各品種以及雜交豬隻之各項統計....................................................................51 表 26. 本試驗A及B兩豬場樣品微衛星標幟交替基因頻率.....................................53 表 27. 大賣場採樣之基因型鑑定結果........................................................................55 表 28. 產銷履歷之驗證結果........................................................................................55 表 29. 單一族群及其內部兩個次族群交替基因頻率及 HE......................................58 | |
dc.language.iso | zh-TW | |
dc.title | 運用微衛星標幟進行台灣豬隻
產銷履歷追溯之驗證 | zh_TW |
dc.title | Using microsatellite markers for the traceability test of fresh pork in Taiwan | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 王佩華(Pei-Hwa Wang) | |
dc.contributor.oralexamcommittee | 宋永義(Yung-Yi Sung),黃木秋(Mu-Chiou Huang) | |
dc.subject.keyword | 猪,微衛星標幟,產銷履歷,個體鑑別, | zh_TW |
dc.subject.keyword | Pigs,Microsatellite markers,Traceability,Individual identification, | en |
dc.relation.page | 73 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-07-27 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
顯示於系所單位: | 動物科學技術學系 |
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