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引进美国SPF杜洛克种猪抗腹泻基因检测及后代选育研究
摘要 为在种猪世代选育中加大GG纯合抗性个体的选留比例,建立杜洛克抗性K88(F4)腹泻专门化品系,试验对引进美国SPF杜洛克种猪后代中的43头(其中母猪37头,公猪6头)进行耳朵组织采样,样本送往江西农业大学动物生物技术国家重点实验室培育基地进行抗腹泻基因检测;之后将已检测过抗腹泻基因的种猪分为2组,试验组按GG(♂)×GG(♀),对照组按GG(♂)×GA(♀)进行配种,并进行后代选育研究。结果发现:杜洛克种猪群GG基因型个体比例高达86.05%,仅有5头GA基因型个体和1头AA基因型个体;在相同的饲养条件下,试验组哺乳种猪腹泻发生率为2.97%、死亡率为0.99%,30日龄断奶仔猪成活率为97.03%,对照组差异明显。
仔猪腹泻是国内外养猪业常见易发、多发疾病,不仅影响仔猪成活率,而且能直接或间接第造成巨大的经济损失。能致哺乳仔猪腹泻的疾病有很多种,其中以仔猪红痢、仔猪白痢、仔猪黄痢最为常见。据统计,目前我国平均每头母猪年生产断奶仔猪只有11~13头,其原因除年产窝数较少(约1.7~1.9窝)、窝产仔猪较少(不足10头)外,仔猪死亡率高(大20%~30%)也是重要原因之一。仔猪因腹泻导致死亡的头数约占总死亡头数的49%,由腹泻导致仔猪生长发育受到影响造成的潜在损失也很大。因此,预防仔猪腹泻,提高仔猪成活率,可以更大限度地发挥养猪业的生产潜能,获得较大的经济效益。本试验对引进美国SPF杜洛克种猪的后代进行了抗腹泻选育研究,以期为良种猪的抗病育种工作及推广应用积累一些数据和经验。
1、材料与方法
1.1试验时间与地点
试验于2012年10月8日至2013年3月1日在贵州省畜禽良种场杜洛克生产区进行。
1.2试验动物
选取引进美国SPF杜洛克种猪后代中的43头(其中母猪37头,公猪6头)进行耳朵组织采样(≥300mg)。样品送往江西农业大学动物生物技术国家重点实验室培育基地进行抗腹泻基因检测。
1.3抗腹泻基因检测
采用SNaPshot法进行基因检测。该技术由美国应用生物公司(ABI)开发,是基于荧光标记单碱基延伸原理的分型技术(也称小测序),主要针对中等通量的SNP分型项目。在一个含有测序酶、4种荧光标记ddNTP、紧临多态位点5′一端的不同长度延伸引物和PCR产物模板的反应体系中,引物延伸一个碱基即终止,经ABI测序仪检测后,根据峰的移动位置确定该延伸产物对应的SNP位点,根据峰的颜色可得知掺入的碱基种类,从而确定该样本的基因型。对于PCR产物模板,可通过多重PCR反应体系来获得。通常用于10~30个SNP位点分析。
1.4后代选育研究
根据抗腹泻基因检测结果,淘汰1头AA基因型的母猪。2012年10月8日,依试验设计要求,试验组按GG(♂)×GG(♀)、对照组按GG(♂)×GA(♀)进行配种。2013年2月1日至3月20日为试验观察期,在相同的饲养条件下,观察并记录哺乳仔猪腹泻的发生和治愈情况、30日龄仔猪断奶成活率等。
2、结果与分析
抗腹泻基因的检查结果
抗腹泻基因的检查结果如表1所示。
   表1  杜洛克种猪抗腹泻基因的检测结果

 注:G为断奶前仔猪F4ac腹泻抗性
Abstract increase in pig breeding generations homozygous GG elected to stay in the proportion of resistant individuals , the establishment of Duroc -resistant K88 (F4) diarrhea specialized strains , the test for the introduction of American Duroc progeny SPF 43 ( which 37 sows , boars 6 ) of the ear tissue samples , samples were taken for animal Biotechnology , Jiangxi Agricultural University, State Key Laboratory Breeding Base for anti- diarrheal genetic testing ; after the gene has been detected anti-diarrhea divided into two groups of pigs test group by GG (♂) × GG (♀), the control group by GG (♂) × GA (♀) be breeding , and breeding of future generations . The results showed that : Duroc group GG genotype as high as 86.05% , only 5 GA genotype and an AA genotype ; reared under the same conditions , the experimental group breastfeeding rate was 2.97% pig diarrhea , mortality rate was 0.99% , 30 -day-old weaned survival rate was 97.03% in the control group significantly different .
Diarrhea is a common domestic pig industry prone , multi- disease not only affects the survival rate of piglets , and can directly or indirectly cause huge economic losses first . Piglets can cause diarrhea diseases there are many, which piglets red dysentery , diarrhea piglets , yellow scour the most common. According to statistics, China's average annual production per sow weaned only 11 to 13 , in addition to an annual litter the reason fewer ( about 1.7 to 1.9 nest ) , nest farrow fewer ( less than 10 ) , the piglets high mortality ( 20% to 30% of large ) is one of the important reasons. Piglet deaths due to diarrhea leading to the head of the head approximately 49 % of the total number of deaths from diarrhea resulting in a potential loss of piglets also greatly affected by the result . Therefore , prevention of diarrhea and improve the survival rate of piglets , you can better maximize the production potential of the pig industry and achieve greater economic efficiency. The trial of the United States to introduce SPF Duroc breeding resistant offspring were studied diarrhea, with a view to breeding disease-resistant varieties of pigs and promote the use of accumulated some data and experience.
1 Materials and methods
1.1 Test time and place
Test on October 8, 2012 to March 1, 2013 carried out in Guizhou Province Duroc breeding farm livestock production areas .
1.2 Test Animals
Select the introduction of American Duroc progeny SPF 43 ( of which 37 sows , boars 6 ) of the ear tissue samples (≥ 300mg). Samples sent to the National Laboratory of Animal Biotechnology , Jiangxi Agricultural University training base for anti- diarrheal genetic testing .
1.3 anti-diarrhea genetic testing
SNaPshot method using genetic testing . The technology was developed by the U.S. company Applied Biosystems (ABI) to develop , is based on the principle of fluorescently labeled single base extension genotyping technology ( also known as small sequencing ) , mainly for medium -throughput SNP genotyping projects. After containing Sequenase , four kinds of fluorescent labeled ddNTP, close to the polymorphic site 5 ' end of different lengths of the primer extension reaction system and a PCR product template, single base primer extension is terminated by ABI sequencing detected , the mobile determines that the peak position of the extension product of the genotype corresponding to SNP sites , a peak based on the color base species that may be incorporated in order to determine the sample . Template for PCR products , by multiplex PCR reaction system to obtain . Typically used for 10 to 30 SNP loci analysis .
1.4 Breeding of offspring
According to the results of genetic testing for anti-diarrheal , out of an AA genotype sows. October 8, 2012 , in accordance with the design requirements of the test , the test group by GG (♂) × GG (♀), the control group by GG (♂) × GA (♀) conducted breeding . 2013年1 February to 20 March for the trial observation period , under the same rearing conditions , observed and recorded the occurrence of breast-feeding and cure diarrhea cases, 30 days post-weaning survival rate and so on.
2 , Results and Analysis
Anti-diarrhea gene test results
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