Genetic analysis of Mexican Criollo cattle populations

The objective of this study was to evaluate the genetic structure of Mexican Criollo cattle populations using microsatellite genetic markers. DNA samples were collected from 168 animals from four Mexican Criollo cattle populations, geographically isolated in remote areas of Sierra Madre Occidental (West Highlands). Also were included samples from two breeds with Iberian origin: the fighting bull (n = 24) and the milking central American Criollo (n = 24) and one Asiatic breed: Guzerat (n = 32). Genetic analysis consisted of the estimation of the genetic diversity in each population by the allele number and the average expected heterozygosity found in nine microsatellite loci. Furthermore, genetic relationships among the populations were defined by their genetic distances. Our data shows that Mexican cattle populations have a relatively high level of genetic diversity based either on the mean number of alleles (10.2-13.6) and on the expected heterozygosity (0.71-0.85). The degree of observed homozygosity within the Criollo populations was remarkable and probably caused by inbreeding (reduced effective population size) possibly due to reproductive structure within populations. Our data shows that considerable genetic differentiation has been occurred among the Criollo cattle populations in different regions of Mexico.     

草原红牛及其杂种牛若干生产性能的微卫星标记研究

选用草原红牛、利木赞与草原红牛杂交后代共计40头作为试验牛群体,以体重、体尺作为衡量牛生长发育的指标,以肉牛线性体型评分方法中的肌肉度线性评分性状和屠宰肉用性状作为衡量牛肉用性能的指标,运用SPSS软件之GLM分析了40头牛21个性状与8个微卫星标记的关系。结果表明,IDVGA55等位基因C(203bp)对体高、十字部高和坐骨端高3个体尺性状有正向效应;BM2113等位基因C(142bp)对腿围、净肉重和净肉率等性状有正向影响;ETH225等位基因A(123bp)对腰角宽具有正向影响;IDVGA46等位基因C(211bp)对5个肌肉度评分性状和4个屠宰肉用性状有负向影响;BM1824等位基因A(171bp)对十字部高、腰厚2个性状有正向效应,等位基因C(179bp)对胸深性状有正向效应;TGLA44等位基因E(221bp)对肉用性状有正向影响。     

草原红牛及其杂交群体中微卫星DNA多态性研究

选用草原红牛及其与利木赞牛的杂交后代66头作为试验牛群体,提取血液及肝脏基因组DNA,设计8对微卫星引物进行PCR扩增,从分子水平上对草原红牛及其杂交群体8个位点的遗传多态性进行微卫星分析。结果表明,在草原红牛中,ETH225、IDVGA2、IDVGA46和IDVGA44等位基因数分别为5、4、3和4,多态信息含量(PIC)分别为0．5420、0．6736、0．5218和0．5750,这4个位点均为高度多态。而另4个位点BM2113、BM1824、IDVGA55和TGLA44等位基因数分别,2、2、2和5,多态信息含量分别为0．3698、0．3604、0．3538和0．4708,属于中度多态性位点。在杂交牛群体中,BM2113、ETH225、IDVGA2、IDVGA46、ID- VGA44、BM1824、IDVGA55和TGLA44这8个位点的等位基因数分别为4、5、4、4、5、4、4和6,多态信息含量(PIC)分别为0．6432、0．5943、0．6593、0．5794、0．7259、0．6121、0．6120和0．6204,杂合度为0．7034,均属于高度多态性位点。这些微卫星位点作为遗传标记应用于草原红牛遗传育种研究之中是可行的。     

Carcass traits and microsatellite distributions in offspring of sires from three geographical regions of Japan.

We proposed that cattle sired by bulls from distinct geographical regions of Japan would differ in their ability to accumulate marbling and that they could be distinguished by differences in microsatellite genotypes. Semen was obtained from six, three, and one sire from the Hyogo, Shimane, and Tottori regions, respectively. Cows and heifers (n = 92) were bred by artificial insemination and raised at the Kyoto University Livestock Research Farm over several generations. The calves (n = 145) were 252 +/- 0.2 d of age (mean +/-SEM) and weighed 164 to 307 kg at the beginning of the finishing phase. Cattle produced from sires from the three regions differed significantly in days on feed (all significant differences P < 0.05). Carcass data were collected from 48, 36, and 19 offspring from the Hyogo, Shimane, and Tottori sires, respectively. There was no difference in slaughter weight (550 +/- 15 kg). Carcasses from Shimane progeny had more muscle than Hyogo cattle. Hyogo and Shimane cattle contained more kidney and 12th-rib fat than Tottori cattle, whereas Hyogo offspring had more 6th-rib fat than Shimane or Tottori offspring. There were no differences across regions in the monounsaturated:saturated fatty acid ratio. The rate of gain of 6th-rib fat for Hyogo progeny (0.033%/d) was significantly greater than that for Shimane or Tattori progeny (both 0.023%/d). Hyogo 12th-rib fat gain was 0.026%/d, which was significantly greater than that for Tottori progeny (0.010%/d). Shimane 12th-rib fat gain was intermediate (0.016%/d) between the other groups. Blood and muscle samples were used for the collection of DNA from 58, 30, and 18 offspring from the Hyogo, Shimane, and Tottori sires, respectively. Samples of the DNA were analyzed for 11 microsatellites. The BM1824 microsatellites for the Tottori progeny were not in Hardy-Weinberg equilibrium because of successive use of Hyogo sires on previous generations. The TGLA227 microsatellites for the Shimane cattle were not in Hardy-Weinberg equilibrium because of selective removal of progeny for slaughter. There were significant differences in allelic frequencies for the BM1824, ETH10, INRA23, and SPS115 and TGLA53 alleles across regions. These data are consistent with the geographical isolation of the different lines of Japanese Black cattle during their development and indicate the superiority of certain groups of sires in the accumulation of intramuscular lipid.     

草原红牛群体中微卫星DNA多态性的研究

选用草原红牛42头作为试验牛群体,经过牛血液基因组DNA的提取、8对微卫星引物的PCR扩增、扩增产物的聚丙烯酰胺电泳分型、UVIBAND计算机凝胶成像分析系统分析各位点等位基因及全部个体的标记基因型、PPAP3.0软件计算基因频率、多态信息含量(PIC)和杂合度等步骤,从分子水平上分析了草原红牛在8个位点的遗传多态性。结果表明,ETH225、IDVGA2、IDVGA46和IDVGA44等位基因数分别为5、4、3和4,多态信息含量分别为0.542 0,0.673 6,0.521 8和0.575 0,这4个位点为高度多态位点。而另4个位点BM2113、BM1824、IDVGA55和TGLA44等位基因数分别2、2、2和5,多态信息含量分别为0.369 8,0.360 4,0.353 8和0.470 8,属于中度多态性位点。8个位点作为遗传标记应用于草原红牛遗传育种研究之中是可行的。     

草原红牛体尺体重性状微卫星标记的研究

研究选用草原红牛、草原红牛与利木赞牛杂交后代共计40头作为试验牛群体，以体尺、体重作为衡量其生长发育指标．利用SPSS软件分析了8个微卫星位点基因标记与体尺性状的关系。结果表明，IDVGA55等位基因C(203bp)对体高、十字部高和坐骨端高三个体尺性状有正效应；BM2113等位基因C(142bp)对腿围性状有正面影响；ETH225等位基因A(123bp)对腰角宽具有正面影响；BM1824等位基因A(171bp)对十字部高性状有正效应，等位基因C(179bp)对胸深性状有正效应；TGLA44等位基因E(221bp)对体重有正面影响。     

平武黄牛血液蛋白质多态性研究

应用醋酸纤维薄膜和垂直板聚丙烯酰胺凝胶电泳技术对四川平武黄牛地方牛种的血红蛋白（Hb)和血清运铁蛋白（Tf)座位多态性进行了分析。结果：平武黄牛Hb座位含有3种基因HbA,HbB和HbC,HbC为优势基因，组成5种表现型，Tf座位有4个等位基因，TfD1和TfE为优势基因，组成10种表现型；与秦川牛，宣汉牛比较，Hb和Tf座位的不同基因及其因型频率都有很大差异。     

Allelic and Genotypic Frequencies of the Leptin Gene Sau3AI-Restriction Fragment Length Polymorphism and Evaluation of Its Association with Age at Puberty in Cattle in the Southwestern United States and Northern Mexico

Leptin is a hormone secreted by adipose tissue that influences appetite, growth, and reproduction. In cattle, the leptin gene contains a Sau3A-restriction fragment length polymorphism (RFLP) within intron II and exon III. This RFLP yields two alleles (A and B) and three genotypes (AA, AB, and BB). Because level of adiposity is important to numerous performance traits, two studies were conducted to assess the potential use of the leptin Sau3AI-RFLP as a selection tool in cattle in the Southwestern United States and Northern Mexico. The objective of the first study was to survey allelic and genotypic frequencies of the Sau3AI-RFLP in breed groups of cattle selected for beef production. High frequencies (≥0.76) of the A allele were observed in cattle that were Bos taurus (Angus and Criollo), Bos indicus-influenced (Brangus, Brahman, F₁-Hereford x Brahman, Santa Cruz, and Santa Gertrudis), and of historic lineage of Bos taurus x Bos indicus (African Mashona). Genotypic frequencies in these breed groups were 0.82 (AA), 0.16 (AB), and 0.02 (BB). The objective of the second study was to evaluate the relationship of the leptin RFLP to age at puberty in heifers. No relationship was detected (P < 0.38) between genotype and age at puberty. The A allele existed in the breed groups evaluated with high frequency, thus negating the ability to assess the relationship of the B allele to age at puberty. Findings do not support use of the leptin Sau3AI-RFLP as a marker in selection strategies for cattle in the Southwestern United States and Northern Mexico.     

2个与HSP70基因连锁的微卫星座位与牛运输应激性状的关联分析

试验探讨了2个与HSP70基因连锁的微卫星座位与牛运输应激性状的关联分析。选择120头12月龄、体重250 kg左右的健康西门塔尔杂种肉牛进行运输应激试验，并根据牛基因组遗传图谱，选择23号染色体上与HSP70基因连锁的微卫星座位BMS468和BM1258检测其在西门塔尔杂种肉牛样本群体中的多态性，采用最小二乘拟合一般线性模型分析微卫星座位与运输应激性状部分指标之间的关联效应。结果显示：微卫星座位BMS468共检测到5个等位基因（128、134、140、146、154 bp），优势等位基因为128和134 bp，有效等位基因数（Ne）、多态信息含量（PIC）和遗传杂合度（He）分别为3.66、0.68和0.73，遗传多态性较高，关联分析显示与运输后7 d平均日增重和发病率显著相关，其中128/128 bp基因型个体的运输后7 d平均日增重最大（P < 0.05），134/128 bp基因型个体的发病率最低（P < 0.05）；微卫星座位BM1258座位共检测到6个等位基因（99、101、103、113、117、119 bp），优势基因为101 bp，Ne、PIC和He分别为4.30、0.73和0.77，遗传多态性较丰富，关联分析显示，其与发病率显著相关，其中99/99 bp基因型个体的发病率最低（P < 0.05）。综上所述，2个微卫星座位可作为牛运输应激性状的潜在遗传标记，为开展牛抗运输应激性状的标记辅助选择提供科学依据。     

Promoter region of the bovine growth hormone receptor gene: single nucleotide polymorphism discovery in cattle and association with performance in Brangus bulls

Expression of the GH receptor (GHR) gene and its binding with GH is essential for growth and fat metabolism. A GT microsatellite exists in the promoter of bovine GHR segregating short (11 bp) and long (16 to 20 bp) allele sequences. To detect SNP and complete an association study of genotype to phenotype, we resequenced a 1,195-bp fragment of DNA including the GT microsatellite and exon 1A. Resequencing was completed in 48 familialy unrelated Holstein, Jersey, Brown Swiss, Simmental, Angus, Brahman, and Brangus cattle. Nine SNP were identified. Phylogeny analyses revealed minor distance (i.e., <5%) in DNA sequence among the 5 Bos taurus breeds; however, sequence from Brahman cattle averaged 27.4 +/- 0.07% divergence from the Bos taurus breeds, whereas divergence of Brangus was intermediate. An association study of genotype to phenotype was completed with data from growing Brangus bulls (n = 553 from 96 sires) and data from 4 of the SNP flanking the GT microsatellite. These SNP were found to be in Hardy-Weinberg equilibrium and in phase based on linkage disequilibrium analyses (r(2) = 0.84 and D'= 0.92). An A/G tag SNP was identified (ss86273136) and was located in exon 1A, which began 88 bp downstream from the GT microsatellite. Minor allele frequency of the tag SNP was greater than 10%, and Mendelian segregation was verified in 3 generation pedigrees. The A allele was derived from Brahman, and the G allele was derived from Angus. This tag SNP genotype was a significant effect in analyses of rib fat data collected with ultrasound when bulls were ~365 d of age. Specifically, bulls of the GG genotype had 6.1% more (P = 0.0204) rib fat than bulls of the AA and AG genotypes, respectively. Tag SNP (ss86273136), located in the promoter of GHR, appears to be associated with a measure of corporal fat in Bos taurus x Bos indicus composite cattle.     

Frequency of bovine Nramp1 (Slc11a1) alleles in Holstein and Zebu breeds

Natural resistance against brucellosis in cattle is linked to the Nramp1 gene, which encodes a divalent cation transporter that localizes in the phagolysosome membrane in macrophages. Nramp1 gene in mouse plays a critical role in innate immunity favoring bacterial killing by macrophages in addition to its influence on adaptative immunity. Polymorphisms at the bovine Nramp1 3' untranslated region (3'UTR), detectable by Single Strand Conformational Analysis (SSCA), are associated with natural resistance against brucellosis. Such polymorphisms are associated with variation in the number of GT repeats. This study compared the frequency of Nramp1 3'UTR polymorphisms between Zebu and European bovine breeds. Eighty-one Holsteins (Bos taurus taurus) and 167 Zebu (Bos taurus indicus), including the following breeds: Nelore (n=95), Guzerá (n=37), and Gir (n=35), totaling 248 pure breed cattle studied. DNA extraction was performed using the guanidium protocol and genotyping was performed by SSCA. DNA from cattle considered genotypically resistant to brucellosis resulted in a single band (homozygous) with 175bp, corresponding to the 3'UTR with 13 GT pairs (GT13), whereas DNA from genotypically susceptible cattle generated one single band with 177bp (homozygous GT14) or double bands with both 175 and 177bp, or 175 and 179bp (heterozygous GT13/GT14 or GT13/GT15, respectively). A marked difference in the frequency of alleles was detected between the Zebu and Holstein cattle. Holsteins had an extremely homogeneous genotype, with 100% of the individuals with a GT13 genotype. In sharp contrast the Nelore breed had the most heterogeneous genotype with four allelic combinations, namely, homozygous GT13, homozygous GT14, heterozygous GT13/GT14, and heterozygous GT13/GT15. When the Zebu breeds were compared to each other, the only significant difference observed was the frequencies of the genotypes GT13 and GT14 between the Nelore and Guzerá breeds. The knowledge of allelic frequencies in different breeds of cattle may prove to be very useful in the future for planning breeding strategies for selection of resistant cattle.     

Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene

The aim of the present study was to identify and characterize polymorphisms within the 5′ flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental × Nelore (n = 30), Canchim × Nelore (n = 30) and Angus × Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648 , AY662649 and AY662650 , respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837 ), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652 , respectively). The allele sequences were compared with GenBank sequences ( M57764 , AF007750 and AH009106 ) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies.     

Four p67 alleles identified in South African Theileria parva field samples

Previous studies characterizing the Theileria parva p67 gene in East Africa revealed two alleles. Cattle-derived isolates associated with East Coast fever (ECF) have a 129 bp deletion in the central region of the p67 gene (allele 1), compared to buffalo-derived isolates with no deletion (allele 2). In South Africa, Corridor disease outbreaks occur if there is contact between infected buffalo and susceptible cattle in the presence of vector ticks. Although ECF was introduced into South Africa in the early 20th century, it has been eradicated and it is thought that there has been no cattle to cattle transmission of T. parva since. The variable region of the p67 gene was amplified and the gene sequences analyzed to characterize South African T. parva parasites that occur in buffalo, in cattle from farms where Corridor disease outbreaks were diagnosed and in experimentally infected cattle. Four p67 alleles were identified, including alleles 1 and 2 previously detected in East African cattle and buffalo, respectively, as well as two novel alleles, one with a different 174 bp deletion (allele 3), the other with a similar sequence to allele 3 but with no deletion (allele 4). Sequence variants of allele 1 were obtained from field samples originating from both cattle and buffalo. Allele 1 was also obtained from a bovine that tested T. parva positive from a farm near Ladysmith in the KwaZulu-Natal Province. East Coast fever was not diagnosed on this farm, but the p67 sequence was identical to that of T. parva Muguga, an isolate that causes ECF in Kenya. Variants of allele 2 were obtained from all T. parva samples from both buffalo and cattle, except Lad 10 and Zam 5. Phylogenetic analysis revealed that alleles 3 and 4 are monophyletic and diverged early from the other alleles. These novel alleles were not identified from South African field samples collected from cattle; however allele 3, with a p67 sequence identical to those obtained in South African field samples from buffalo, was obtained from a Zambian field isolate of a naturally infected bovine diagnosed with ECF. The p67 genetic profiles appear to be more complex than previously thought and cannot be used to distinguish between cattle- and buffalo-derived T. parva isolates in South Africa. The significance of the different p67 alleles, particularly the novel variants, in the epidemiology of theileriosis in South Africa still needs to be determined.     

Analysis of Two Chinese Yak(Bos grunniens) Population Using Bovine Microsatellite Primers

Two Chinese domestic yak populations representing the Plateau type and the Huanhu Alpine type were analysed with 12 bovine microsatellite primers. All primer pairs functioned in the yak genome and polymorphism was found at all loci. The allele size ranges and frequencies of the two yak populations were similar and there was considerable overlap with the allele size ranges observed in cattle. Data for European cattle breeds was obtained from the Cattle Diversity Database(CaDBase)to interpret the heterozygosity and genetic distance estimates in yak populations. Heterozygosity estimated for the two yak populations was comparable to that of European cattle while Nei's Genetic Distance DA between the two yak populations was less than distances between the most closely related German cattle breeds. Bovine microsatellite primers proved to be a valuable tool for characterization of yak populations.     

胰岛素样生长因子Ⅰ基因第1内含子微卫星多态性对山西白猪体重和体尺性状的影响

试验检测了山西白猪174个个体胰岛素样生长因子-Ⅰ(insulin-like growth factor Ⅰ,IGF-Ⅰ)基因第1内含子以CA为核心重复序列的微卫星座位的多态性,并利用单变量动物模型分析了多态位点对山西白猪不同阶段体重和体尺性状的影响。结果表明,在该座位上,共检测到6个等位基因,分别为198、202、204、206、208和210 bp,表现出CA重复数较低的等位基因198和202 bp频率较高,分别为0.2759和0.2385;CA重复数较高的等位基因208 bp频率较低,为0.0747;共检测到12种基因型,纯合子频率较高,均大于0.010(208/208 bp类型除外);而杂合子个体频率都较低,为0.017。统计分析结果表明,IGF-Ⅰ基因型对山西白猪群体的初生重、断奶重、6月龄体重和体高、体长、胸围、背膘厚等性状有显著影响(P＜0.05)。204/204 bp基因型个体不同生长阶段体重高于其他类型个体,而6月龄活体背膘厚又显著低于其他类型个体,符合中国当前猪育种方向,在选育中应优先选留。     

Frequencies of PRNP gene polymorphisms in Vietnamese dairy cattle for potential association with BSE

Summary Since 2004, significant associations between bovine spongiform encephalopathy (BSE) susceptibility in cattle and frequencies of insertion/deletion (ins/del; indel) polymorphisms within the bovine prion protein gene (PRNP) have been reported. In this study, we investigated the frequencies of indel polymorphisms within two variable sites, a 23-bp indel polymorphism in the promoter region (23indel) and a 12-bp indel polymorphism in intron 1 region (12indel), in the PRNP in 206 Vietnamese dairy cattle and seven Japanese BSE-affected cattle. In Vietnamese dairy cattle, the frequency distributions of del allele and del/del genotypic polymorphisms in the 23indel site, which are thought to be associated with BSE susceptibility, were significantly higher, whereas the frequencies of del allelic and del/del genotypic polymorphisms in the 12indel site, which have been reported to confer BSE susceptibility, were significantly lower. We have provided evidence that Vietnamese dairy cattle have a unique genetic background in the PRNP gene in comparison with cattle or sires previously reported in other countries.     

南阳牛IGF-2基因PCR-RFLP分析

利用PCR-RFLP技术和限制性内切酶Bsr I对85头南阳牛类胰岛素样因子2(IGF-2)进行了分析。结果表明：南阳牛在该位点检测到两种等位基因A和B,其基因频率分别为0．5471和0．4529。A等位基因包括185bp和32bp两个片段,B等位基因包括118bp、67bp、32bp三个片段。南阳牛在该位点处于Hardy-Weinberg平衡状态。     

临夏黄牛某些种质特征的研究报告

采用中心产区系统随机整群抽样法，对监夏黄牛两个系统五个群体共１８１头牛进行了遗传检测，描述了临床夏黄牛的体型外貌特征，体尺特以及毛色特征等，利用聚丙烯酰胺凝胶电沪法对血红蛋白，白蛋白，转铁蛋白，后转铁蛋白－１，碱性磷酶等个血液蛋白多态位点进行了分析。     

草原红牛肉用性能微卫星标记的研究

选用草原红牛、利木赞牛与草原红牛杂交后代共计40头作为试验牛群体，以内牛线性体型评分方法中的肌肉度评分性状和屠宰肉用性状作为衡量其肉用性能的指标，利用SPSS软件分析了8个微卫星位点基因标记与肉用性能的关系。结果表明，BM2113等位基因C（142bp）对净肉重和净肉率性状有正面影响；IDVGA46等位基因C（211bp）对内用性能有负面影响；BM1824等位基因A（171bp）时腰厚性状有正效应；TGLA44等位基因E（221bp）对髻甲、腰厚、臀部外形等肌肉度评分性状和体质量、胴体重以及净肉重等屠宰肉用性状有正面影响。