Using microsatellite markers for the traceability test of fresh pork in Taiwan
Date Issued
2009
Date
2009
Author(s)
Lyu, Yue-Jheng
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.n 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. he 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.n 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.
Subjects
Pigs
Microsatellite markers
Traceability
Individual identification
SDGs
Type
thesis
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