A 1.1 Mb duplication CNV on chromosome 17 contributes to skeletal muscle development in Boer goats

The Boer goat is one of the top meat breeds in modern animal husbandry and has attracted widespread attention for its unique growth performance.However,the genetic basis of muscle development in the Boer goat remains obscure.In this study,we identified specific structural variants in the Boer goat based on genome-wide selection signals and analyzed the basis of the molecular heredity of related candidate genes in muscle development.A total of9 959 autosomal copy number variations(CNVs) were identified through selection signal analysis in 127 goat genomes.Specifically,we confirmed that the highest signal CNV(HSV) was a chromosomal arrangement containing an approximately 1.11 Mb(CHIR17:60062304-61171840 bp) duplicated fragment inserted in reverse orientation and a 5 362 bp deleted region(CHIR17:60145940-60151302 bp) with overlapping genes(e.g.,ARHGAP10,NR3C2,EDNRA,PRMT9,and TMEM184C).The homozygous duplicated HSV genotype(+/+) was found in 96% of Boer goats but was not detected in Eurasian goats and was only detected in 4% of indigenous African goats.The expression network of three candidate genes(ARHGAP10,NR3C2,and EDNRA)regulating dose transcription was constructed by RNA sequencing.Results indicated that these genes were involved in the proliferation and differentiation of skeletal muscle satellite cells(SMSCs) and their overexpression significantly increased the expression of SAA3.The HSV of the Boer goat contributed to superior skeletal muscle growth via the dose effects of overlapping genes.

Identification and Phylogenetic Analysis of an Orf Virus Isolated from an Outbreak in Boer Goat in Shanxi Province

To identify and analyze the Orf virus in Shanxi Province, China, an Orf virus strain was successfully isolated from crust materials of boer goat with clinical sore mouth symptom from a goat farm of Shanxi Province by passaging in lamb testis （LT）. The Orf virus was identified by enzyme linked immunosorbent assay （ELISA） test, recurrent infection test, transmission electron microscopy, and PCR. The nucleotide and amino acid sequences of two genes of the Orf virus were analyzed. The results showed that under the electron microscopy the virus had a presence of typical parapoxvirus virions and there were many eosinophilic intracytoplasmic inclusions observed by hematoxylin-eosin （H＆E） stain. In ELISA test, optical density （OD） readings of the sample showed a positive result, and the rabbits infected with the virus showed a typically Orf virus-infected appearance. All these findings proved that the sample was an Orf virus. The phylogenetic studies of Orf B2L and Orf F1L genes showed that the virus clustered in different branches and were closer to the Orf virus Nantou （DQ904351） and the OV-SA00 isolates （AY386264）. Furthermore, the above results may provide some insight into the genotype of the etiological agent responsible for the Orf outbreak in Shanxi Province, and could also provide a comparative view of the B2L and F1L genes of parapoxvirus.

Early Growth and Development of Boer Goats from Embryo Transplant

[ Objective] To study the growth-development of Boer goats from embryo transplant. [ Method] With Boer goats from embryo transplant as the tested animals and natural breeding Boer goats born in the same period as CK, the weight of Boer goats at birth and at the age of 1,3 and 6 months as well as the body size at the age of 3 and 6 months were investigated in this study, while their health conditions were also observed and recorded. [ Result] Under the normal feeding conditions, Boer goats from embryo transplant developed well at early stage, and their meat-using body type were also obvious. The average birth weight of bucks was （4.25±0.95） kg, and that of does was （3.74±0.10） kg. The average weight of bucks at the age of 6 months was （31.90 ±0.74） kg and that of was （25.90 ±0.67） kg. The daily gains of bucks and ewes from birth to 6 months of age were 153.64 and 123.11 g respectively, while their body length, body height and chest circumference increased obviously, and there was no significant difference in body weight or body size between them and natural breeding goats. [ Condusioa] The study lays a foundation for further studies on the breedinq and utilization of Boer noats from embryo transplant at early staae.

AFLP and RAPD Analysis of the Boer and Indigenous Breeds of the Goat in Jiangsu

Blood and tissue samples were collected from 105 goats including 60 Boer goats (30 for each sex), 30 Xuhuai goats (15 for each sex) and 15 Haimen goats (7 stud and 8 does). DNA was extracted and DNA pools were constructed on the basis of goat breeds. In 36 selective primer combinations, 29 combinations amplified totally 3 253 markers including 92 polymorphic markers by amplified fragment length polymorphism (AFLP). On average, 3.17 polymorphic markers were amplified per combination, with a polymorphic frequency of 2. 8%. The primer combinations amplifying more polymorphic markers (showed in brackets) were involved in E00+ACG/M00+CAA (13), E00+ACG/M00+CAG (10), E00+AAC/M00+CAC (8) and E00+A AC/M00+ACT (7). A total of 183 markers including 60 polymorphic markers were amplified by RAPD from the pooled DNA of three breeds using 22 primers with strong polymorphism and high reproduc-ibility selected from 93 RAPD primers. On average, 2. 73 polymorphic markers were amplified per primer, with a polymorphic frequency of 32.8%. The results of AFLP and RAPD coincidently suggested that the genetic distance is the closest between Xuhuai and Haimen goat, next between Xuhuai and Boer goat, and the farthest between Haimen and Boer goat. According to the UPGMA method, Haimen and Xuhuai goats can be gathered together as a cluster, then Boer goat. Both methods can be used to implicate the genetic difference of these three breeds, in particular AFLP has more polymorphic markers.

长江三角洲白山羊不同杂交组合早期生长发育性状研究

为了研究长江三角洲白山羊不同杂交组合生长发育的差异,以长江三角洲白山羊为母本、以萨能奶山羊和波尔山羊为父本进行杂交选育,分析萨白F_(1)、波萨白F_(2)和长江三角洲白山羊F_(0)60日龄体重与体尺指标的变化。结果显示:波萨白F_(2)各日龄体重、平均日增重均显著高于长江三角洲白山羊F_(0)(P<0.05);波萨白F_(2)各日龄体重和平均日增重均高于萨白F_(1),萨白F_(1)各日龄体重和平均日增重均高于长江三角洲白山羊F_(0),但差异均不显著(P>0.05)。整个试验期间,波萨白F_(2)、萨白F_(1)各日龄体尺指标均高于长江三角洲白山羊F_(0),但差异均不显著(P>0.05);各组羊的体尺数据随着日龄的增加呈递增的趋势。该试验通过分析不同杂交组合羔羊体重、体尺等早期生长发育性状,为培育具有较高经济价值的长江三角洲白山羊肉用品种(系)提供了一定参考。

姜黄素对热应激状态下波槐杂交羊生长性能和血液生化指标的影响

本研究旨在评价不同添加剂量的姜黄素对热应激条件下波槐杂交羊生长性能及血液生化指标的影响。试验将30头体重为(28.03±0.32)kg的健康波槐杂交公羊随机分为3组,每组10只,分别饲喂0 mg/头/d(CON组)、450 mg/头/d(CUL组)、900 mg/头/d(CUH组)的姜黄素,预饲期15 d,正试期60 d。结果显示:(1)CUL组波槐杂交羊的平均日增重和总增重显著高于CON和CUH组(P<0.05)。(2)CUL组波槐杂交羊血清中的总蛋白和球蛋白含量显著高于CON和CUH组(P<0.05)。(3)CUL组杂交羊血清中的免疫球蛋白G含量显著高于CON和CUH组(P<0.05),CON和CUH组杂交羊血清中的丙二醛含量显著高于CUL组(P<0.05)。综上所述,日粮中添加450 mg/头/d的姜黄素可以改善热应激条件下波槐杂交羊的应激状态和健康状态,并进一步提高生长性能。

Genetic Polymorphism of Ten Microsatellites in Two Goat Breeds and Its Relationship with Heterosis

[Objective] This study aimed to investigate the genetic diversity of Macheng black goat and its correlation with heterosis.[Method] Ten microsatellite markers were selected for polymorphism investigation and statistical analysis of Boer goat and Macheng black goat populations.[Result] The results showed that totally 175 alleles were found in 10 microsatellite loci; to be specific,the maximum number of detected alleles was 23,and the minimum number was 10; the effective number of alleles （Ne） was 6.4-18.1,with absolute difference value of 1.6-8.1 from the observed number of alleles.The highest gene frequency was 0.239 1 and the lowest was 0.002 7.The polymorphic information contents of all the ten microsatellite markers were above 0.95.The observed heterozygosity （Ho） ranged from 0.616 7 to 0.984 4 and the expected heterozygosity （He） ranged from 0.844 1 to 0.944 6.The average expected heterozygosity of Boer goat and Macheng black goat was respectively 0.894 0 and 0.906 7.Various body weight and body size indices of Boer goatxMacheng black goat hybrids were improved in varying degrees compared with Macheng black goat （with an increase range of 0.32％-30.06％）.The average heterosis rates of body height and chest girth were relatively high,while average heterosis rate of body weight was relatively low.[Conclusion] The genetic distance between Boer goat and Macheng black goat was 0.379 5,which is consistent with the geographical distribution of Boer goat and Macheng black goat populations and is fully relevant to the heterosis of Boer goat × Macheng black goat hybrids,indicating that investigating polymorphism via microsatellite loci is one of the feasible means to predict and analyze heterosis between varieties.

不同月龄波尔山羊睾丸中精原干细胞活性及增殖特征研究

[目的]探究不同月龄波尔山羊睾丸中精原干细胞(spermatogonial stem cells,SSCs)在体外培养过程中的活性及增殖特征。[方法]分别采集1、3、5、58月龄的波尔山羊睾丸,通过组织学方法分别观察不同月龄波尔山羊睾丸的发育情况;分别收集不同月龄波尔山羊睾丸组织,利用三步酶消化法分离成单细胞,通过台盼蓝染色观察睾丸消化细胞总数和死亡细胞数;采用0.2%明胶富集差速贴壁法纯化SSCs并培养10 d,通过流式细胞术鉴定细胞悬液纯化前、纯化后及培养10 d后的Thy-1阳性(Thy-1+)细胞数;将纯化的SSCs培养10 d后,采用碱性磷酸酶染色鉴定SSCs克隆,分别计算不同月龄波尔山羊睾丸SSCs形成的克隆数和克隆面积。[结果]睾丸组织学观察发现1月龄波尔山羊曲细精管管腔生殖细胞数量最少,随着波尔山羊月龄的增加,逐渐发育出各级生殖细胞,但58月龄波尔山羊睾丸生精上皮明显退化。不同月龄波尔山羊睾丸组织经过三步酶消化,在纯化前及纯化后获得的睾丸细胞总数以5月龄波尔山羊最高,显著(P<0.05)高于1月龄波尔山羊;不同月龄波尔山羊纯化前及纯化后的睾丸细胞存活率之间均无显著(P>0.05)差异。在纯化前、纯化后的睾丸组织细胞以及培养10 d的纯化SSCs中,Thy-1+细胞数占睾丸细胞总数的比例均以1月龄波尔山羊最高,显著(P<0.05)高于其他月龄波尔山羊。培养10 d后,不同月龄波尔山羊睾丸来源的SSCs克隆均表现为碱性磷酸酶阳性,1月龄波尔山羊睾丸SSCs形成的平均克隆数显著(P<0.05)高于其他月龄波尔山羊,不同月龄波尔山羊的SSCs平均克隆面积无显著(P>0.05)差异。[结论]5月龄波尔山羊睾丸中细胞总数最多,1月龄波尔山羊睾丸中细胞总数最少,但SSCs活性和增殖效率最高。

Boer山羊精子体外获能前后超微结构的研究

用S培养体系对Boer山羊精子进行体外获能试验,借助电子显微技术观察其获能前后超微结构的变化.结果表明,Boer山羊精子的超微结构与其它哺乳动物的类似,分头、颈、尾3部分。头部主要由细胞核构成,含有遗传信息;颈部脆弱,尾部易从此脱落;尾部是精子运动的器官,由中段、主段和末段3部分组成.精子获能后,顶体质膜膨胀、断裂和丢失;顶体外膜囊泡化,进而顶体内膜裸露.

日粮添加不同水平的氧化锌对波尔山羊卵巢活动和繁殖性能的影响

本试验将不同水平的氧化锌添加到波尔山羊的日常饲养中,研究其对卵泡活动和繁殖性能的影响。试验将60只山羊平均分为4组,每组3个重复,每个重复5只羊。分别为对照组、试验I组(日粮+300mg/kg氧化锌)、试验Ⅱ组(日粮+500mg/kg氧化锌)和试验Ⅲ组(日粮+700mg/kg氧化锌)。试验为期40d,试验结束后测定其卵泡活动、繁殖激素和繁殖性能等指标。结果发现,与对照组相比,在日粮中添加不同水平的氧化锌对波尔山羊的促性腺激素释放激素(GnRH)、促卵泡成熟激素(FSH)、促黄体生成激素(LH)均有显著影响(P <0.05),其中添加500mg/kg氧化锌对提升波尔山羊的卵泡活动效果最显著(P <0.05),GnRH、FSH和LH比对照组分别提高18.17%、29.34%和27.14%;其次,在日粮中添加不同水平的氧化锌对波尔山羊血清中的雌二醇(E2)、孕酮(P4)等繁殖激素指标也有显著影响(P <0.05),其中试验Ⅱ组的繁殖激素指标与对照组相比差异显著(P <0.05),E2、P4分别提高11.56%和27.93%。最后,在日粮中添加不同水平的氧化锌对波尔山羊的繁殖性能也有显著影响(P <0.05),其中500mg/kg氧化锌试验组羊受胎率、产羔率及双羔率与对照组相比差异显著(P <0.05),受胎率、产羔率及双羔率分别比对照组提高30.8%、33.3%和33.3%。研究结果表明,在日粮中添加500mg/kg的氧化锌,对波尔山羊的卵泡活动和繁殖性能的促进效果最显著。

布尔(Boer)山羊种质特性的研究——生理常值及血液生理生化

对布尔山羊生理常值及血液生理生化指标测定结果表明，布尔山羊除羔羊淋巴细胞显著高于成年羊（Ｐ＜００１）外，其余各项指标在年龄间、性别间均无显著差异（Ｐ＞００５）。该品种这些生理生化指标与其它山羊的基本接近或在正常变化范围内。

配种前饲喂不同能量和蛋白水平的日粮对波尔山羊繁殖性能的影响

文章旨在研究波尔山羊在配种前饲喂不同蛋白水平和能量的日粮对其繁殖性能的影响。试验将39头4月龄的健康波尔母山羊分为3组,各13只,试验1组采用高能高蛋白日粮;试验2组采用中能中蛋白日粮;对照组采用低能低蛋白日粮,对比三组波尔山羊繁殖性能差异。结果显示,三组母波尔山羊初始体重与饲养10 d的差异不显著(P>0.05);饲养50 d与饲养100 d平均日增长体重与背膘厚组间差异与组内差异均显著(P<0.05);试验1组总产仔数量、产仔成活率、初窝幼仔平均体重与其他两组差异显著(P<0.05);三组波尔山羊在35和80 d测量的CAT指标和MDA指标均差异显著(P<0.05)。所以认为采用高能高蛋白日粮对波尔山羊繁殖性能有更优的效果,可以在实际饲养过程中推广使用。

波尔山羊ACVRⅡB基因5′调控区多态性及其与初生体重体尺的关联分析

旨在探讨波尔山羊激活素受体ⅡB(activin A receptor typeⅡB,ACVRⅡB)基因5′调控区多态性及其对初生体重体尺的影响。从103只波尔山羊耳组织中提取基因组DNA,扩增ACVRⅡB基因5′调控区序列,测序鉴定SNPs位点,分析遗传多态性,并与波尔山羊初生体重、体尺进行关联分析。结果显示,鉴定到g.-1776C>G、g.-1725G>A、g.-1697C>T、g.-1604G>A、g.-1569G>C 5个SNPs位点,其中g.-1725G>A位点AA型在群体中频率为0.049,AG型为0.359,GG型为0.592,PIC为0.29,处于中度多态;g.-1569G>C位点CG型在群体中频率为0.167,GG型为0.833,PIC为0.14,处于低度多态。连锁分析发现g.-1725 G>A与g.-1697C>T、g.-1569G>C完全连锁(相关性为100%)。关联分析发现,g.-1725G>A位点AG型初生重、初生体长均显著高于GG型(P<0.05);g.-1569G>C位点CG型初生体高显著高于GG型。综上,g.-1725G>A位点可作为波尔山羊早期生长选育的候选分子标记。

波尔山羊杂交后代及其亲本随机扩增多态DNA研究

利用随机扩增多态DNA技术研究了波尔山羊、唐山奶山羊、青龙本地山羊以及波尔山羊与这两个山羊群体杂交后代共计128个山羊个体的随机扩增多态DNA。结果表明 :(1)总群体平均遗传多样性指数 (Hsp)为0 6974,群体遗传分化指数为0 9706,山羊群体间平均遗传距离指数 (0 1314～0 2052)明显大于群体内的相应值 (0 0582～0 1440) ,上述结果说明 ,所研究山羊群体不仅具有较为丰富的遗传多样性 ,而且其核基因组遗传变异主要存在于群体间。(2)山羊群体间的分子聚类关系与各群体间的亲缘关系基本一致。

波尔山羊及黄牛耳部成纤维细胞的体外培养

为了获得丰富的核移植供核体细胞 ,通过活体采集波尔山羊及黄牛耳部皮肤 ,利用组织块和消化法开展原代培养 ,并进行传代培养和培养细胞的冷藏、冷冻保存实验。建立了简易的波尔山羊和黄牛成纤维细胞体外培养程序。

苜蓿青饲对波尔山羊屠宰性状及肉品质的影响

选择45只3月龄、体重(18.00±1.01)kg的波尔山羊,根据饲粮中苜蓿鲜草不同的添加比例,随机分为5个处理:对照组(0%)、试验Ⅰ组(15%)、试验Ⅱ组(30%)、试验Ⅲ组(45%)和试验Ⅳ组(60%),6月龄时各处理组随机选择体重中等的波尔山羊3只(♂)进行屠宰,观测对其屠宰性状和肉品质的影响。结果显示,1)各试验组胴体重、屠宰率、净肉率及骨重均高于对照组,各项指标均以试验Ⅰ、Ⅱ组较高,但各试验组与对照组之间各项指标差异并不显著(P>0.05);2)与对照组及其他试验组相比,试验Ⅱ组胸肌的pH值(P<0.01)、肌肉的剪切力值和肌纤维直径、硬脂酸(P<0.05)的含量均最低,而粗脂肪(P<0.05)、蛋白质、亚麻酸(P<0.01)、亚油酸(P<0.05)的含量均最高,对照组的2类指标表现出最高和最低;3)与对照组及其他试验组相比,试验Ⅱ组肌肉中SOD的含量最高,MDA含量最低。以上结果表明,添加新鲜苜蓿能够提高山羊的屠宰性能,还明显降低了波尔山羊肉的膻味,提高了其风味及抗氧化性能,最适宜的添加比例为30%。

波尔山羊胚胎移植技术的研究与应用

对 75只波尔山羊超排处理 ,共获 A、B级胚胎 776枚 ,平均 (10 .35± 6 .4 5 )枚 /只 ,鲜胚移植受体 36 7只 ,移植产羔率达 5 5 .6 %。应用 CIDR和 PMSG同期发情处理受体山羊 5 6 0只 ,5 4 9只达到同期发情 ,32 h内同期率为 98.0 4 %。囊胚和扩张囊胚在发情后 7d的移植产羔率分别为 6 2 .2 2 % (84 /135 )和 5 8.2 6 % (6 7/115 ) ,显著高于桑椹胚 (43.18% )和孵化囊胚 (5 1.72 % ) (P<0 .0 1) ,而囊胚和扩张囊胚间、桑椹胚和孵化囊胚间差异不显著 (P>0 .0 5 )。受体山羊的黄体数量直接影响到移植产羔结果 ,黄体数≥ 2的受体山羊移植产羔率显著高于黄体数为 1的受体山羊移植产羔率 (P<0 .0 1) ,分别为 6 4 .83% (94 /14 5 )和 4 9.5 5 % (110 /2 2 2 )。PMSG注射时间 (在取 CIDR前 2 d或当天注射 )和剂量对受体羊的黄体数和移植产羔率均无显著影响 (P>0 .0 5 )

波尔山羊和江苏本地山羊的AFLP和RAPD分析

实验山羊共 10 5只 ,其中波尔山羊 6 0只 (公母羊各半 ,采血样 2 0个 ,组织样 4 0个 ) ,徐淮山羊 30只 (公母羊各半 ,每羊采血样 1个 ) ,海门山羊 15只 (公羊 7只 ,母羊 8只 ,每羊采血样 1个 )。分别提取DNA ,按品种构建DNA池。应用 36个选择性引物组合对波尔山羊、徐淮山羊和海门山羊品种间AFLP多样性进行研究 ,其中 2 9个引物组合共扩增出 32 5 3个标记 ,包括多态标记 92个 ,平均每个引物组合扩增 3.17个多态标记 ,多态频率达2 .8%。多态标记数较多的引物组合为 :E0 0 +ACG/M 0 0 +CAA(13个 )、E0 0 +ACG/M0 0 +CAG(10个 )、E0 0 +AAC/M0 0 +CAC(8个 )和E0 0 +AAC/M0 0 +ACT(7个 )。从 93个随机引物中筛选出品种间多态性较强、重复性好的引物 2 2个 ,对 3个品种池DNA进行RAPD扩增 ,共扩增出 183个标记 ,其中多态标记 6 0个 ,平均每个引物扩增2 .73个多态标记 ,多态频率为 32 .8%。两种方法得到一致结果 ,即徐淮山羊与海门山羊遗传距离最近 ,徐淮山羊与波尔山羊次之 ,海门山羊与波尔山羊遗传距离最远。按UPGMA法将海门山羊与徐淮山羊聚为一类 ,其次为波尔山羊。两种方法均能较好地反映 3个山羊品种的遗传差异 ,但AFLP方法多态标记数更多。

苜蓿青饲对波尔山羊生长性能及血液生理生化指标的影响

近年来,随着苜蓿(Medicago sativa L.)产业的发展,相关草产品在奶牛饲养中得到广泛应用并日益成熟,但在山羊饲养的消化生理、营养代谢及利用方面的研究却相对缺乏。为了解不同水平苜蓿鲜草饲粮对波尔山羊生长性能及血液生理生化指标的影响,于2005年7-9月,选用3月龄、初始体重17.77±1.05 kg、遗传背景基本一致的波尔山羊(Capra hircus L.)45只,随机分为5个处理组,各组饲粮中苜蓿鲜草替代水平分别为0%(对照组)、15%(试验Ⅰ组)、30%(试验Ⅱ组)、45%(试验Ⅲ组)和60%(试验Ⅳ组)(按干物质计)。结果表明:试验Ⅰ、Ⅱ、Ⅲ组平均日增重和料重比都优于对照组,但差异不显著;试验Ⅱ组血液总蛋白和球蛋白含量极显著(P<0.01)、白蛋白含量显著(P<0.05)高于对照组,而总胆固醇、高密度酯蛋白胆固醇含量显著低于对照组(P<0.05),尽管各组血糖、尿素氮、甘油三酯及低密度酯蛋白胆固醇含量差异不显著,但试验Ⅱ组甘油三酯和低密度酯蛋白胆固醇含量均最低。因此,30%替代水平为最佳,建议在生产中应用。

波尔山羊MyoG基因的鉴定和序列分析

为了探明山羊MyoG基因的序列结构及其对肌肉的分化调控机制,用PCR产物直接测序的方法获得了波尔山羊MyoG基因2363bp长的DNA序列,并对该序列进行了分析。结果表明,波尔山羊MyoG基因包括3个外显子、2个内含子及部分5′UTR(74bp)和3′UTR(260bp)区,编码序列长675bp,共编码224个氨基酸。结构分析表明,该序列所编码的肽链没有信号肽,第1～138个氨基酸为波尔山羊MyoG基因的bHLH结构域。通过比对分析波尔山羊与已知的GenBank中的人类及其它物种MyoG基因发现,该基因编码区核苷酸以及推测的氨基酸同源性和物种间的亲缘关系相一致,无根系统进化树的聚类结果与这些物种本身的生理特性以及传统分类学中已知的生物分类结果相一致。波尔山羊MyoG基因的鉴定及序列分析为山羊肌肉发育调控的机理以及肉质改良等的研究提供了很好的生物学基础信息。