牛卵泡抑素基因克隆及真核表达载体构建

[目的]克隆牛的卵泡抑素基因（Follistatin,FSTN）基因,构建真核表达载体。[方法]用Trizol法从牛的卵巢中提取总RNA,反转录成cDNA,用带有酶切位点牛FSTN的特异性引物扩增其完整编码区序列,连接到T载体、测序,序列无误后亚克隆入真核表达载体pIRES2-AcGFP1中,酶切及PCR鉴定载体。[结...     

Monitoring of the intra-dermal tuberculosis skin test performed by Belgian field practitioners

The present study aimed to monitor skin test practices as performed by veterinarian field practitioners in Belgium. For this purpose, an anonymous postal questionnaire was elaborated and dispatched to veterinarians involved in bovine tuberculosis detection. The questionnaire included items focusing on the skin test performance. International experts in the field of bovine tuberculosis were asked to fill the questionnaire and a scoring scale was built as follows: 0 = ‘ideal’ answer, 1 = acceptable answer, whereas 2 = unacceptable answer. Furthermore, experts were asked to rank the questionnaire’s items according to their possible impact on the risk of not detecting reactors. A global score was further calculated for each participant and a comparison of practices was carried out between the two regions of the country, i.e. Wallonia and Flanders. Significant differences were observed between both regions, a harmonization at the country level is thus essential. No veterinarian summed a null score, corresponding to the ideal skin test procedure, which suggests that skin-testing is far from being performed correctly. Field practitioners need to be sensitized to the importance of correctly performing the test. The authors recommend the questionnaire is suitable for application in other countries or regions.     

Analysis of the spatial variation of Bovine tuberculosis disease risk in Spain (2006-2009)

In this study we explored the spatial variation of Bovine tuberculosis (BTB) risk of being positive, new positive or persistently positive, as well as the risk of eliminating BTB in positive herds throughout Spain from 2006 to 2009 by means of hierarchical Bayesian models. The results of the models showed that the risk of infection (positive or new positive herds), persistence and elimination was lower in counties located in north and north-eastern of Spain, and in the Balearic and Canary islands than in the rest of the country. In some counties the risk of positivity was high during the four years of study, whereas there were others where the risk of positivity was high only in some of the years. With regard to the risk of persistence of BTB positive herds, counties located in the central, western and south-western part of the country had a higher risk in the three studied periods. This study has identified some specific areas of increased BTB risk in Spain, information that is useful for disease management.     

中国荷斯坦种公牛BLAD遗传缺陷的分子检测及系谱分析

本试验运用限制性片段长度多态性聚合酶链式反应（RFLP-PCR）方法,检测我国荷斯坦种公牛白细胞粘附缺陷（bovine leukocyte adhesion deficiency,BLAD）基因的携带频率。共检测了来自全国14个公牛站的587头种公牛,发现BLAD携带者8头,携带率为1.36%。对现有公牛系谱信息分析显示,携带者公牛来自美国、加拿大和中国,其中6头携带者公牛可以追溯到共同祖先Osborndale Ivanhoe。此外,本研究还对我国荷斯坦牛遗传缺陷的控制和携带者公牛的利用提出建议。     

牛呼吸疾病综合征及其防治

牛呼吸疾病综合征是严重危害国内外肉牛养殖业的一种重要传染病,该病的致病因素复杂,包括支原体、病毒和细菌等特异性病原以及运输应激等多种因素。美国等发达国家对该病的攻关研究开始于上世纪80年代初,其研究和临床防控均已达到较高水平,但仍认为该病将是未来10～20年养牛业所面临的主要疾病。我国对该病的认识和研究尚处于起步阶段,借鉴世界上发达国家的先进经验将有助于提升我国对该病的防控水平。     

牛病毒性腹泻病毒E2基因生物信息学分析及可溶性表达

为了获得牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)E2基因的最佳可溶性蛋白,本试验根据GenBank已公布的BVDV E2基因序列,利用DNAStar等软件进行生物信息学分析,截取抗原性和亲水性较高的序列进行稀有密码子优化,串联信号肽序列后合成并连接到原核表达载体pET-32a(+),构建重组质粒pET-32a-Opti-E2;转化至E.coli BL21(DE3)宿主菌,经SDS-PAGE和Western blot分析,成功获得大小为43 kDa的BVDV E2基因的最佳可溶性蛋白,且该蛋白特异性较好。本试验E2蛋白的成功表达可为后续ELISA方法的建立和亚单位疫苗的开发奠定基础。     

Genome-wide detection of signatures of selection in three Valdostana cattle populations

The Valdostana is a local dual purpose cattle breed developed in Italy. Three populations are recognized within this breed, based on coat colour, production level, morphology and temperament: Valdostana Red Pied (VPR), Valdostana Black Pied (VPN) and Valdostana Chestnut (VCA). Here, we investigated putative genomic regions under selection among these three populations using the Bovine 50K SNP array by combining three different statistical methods based either on allele frequencies (F_ST) or extended haplotype homozygosity (iHS and Rsb). In total, 8, 5 and 8 chromosomes harbouring 13, 13 and 16 genomic regions potentially under selection were identified by at least two approaches in VPR, VPN and VCA, respectively. Most of these candidate regions were population-specific but we found one common genomic region spanning 2.38 Mb on BTA06 which either overlaps or is located close to runs of homozygosity islands detected in the three populations. This region included inter alia two well-known genes: KDR, a well-established coat colour gene, and CLOCK, which plays a central role in positive regulation of inflammatory response and in the regulation of the mammalian circadian rhythm. The other candidate regions identified harboured genes associated mainly with milk and meat traits as well as genes involved in immune response/inflammation or associated with behavioural traits. This last category of genes was mainly identified in VCA, which is selected for fighting ability. Overall, our results provide, for the first time, a glimpse into regions of the genome targeted by selection in Valdostana cattle. Finally, this study illustrates the relevance of using multiple complementary approaches to identify genomic regions putatively under selection in livestock.     

Introduction and elimination of Bovine Viral Diarrhoea Virus in a commercial beef herd: a case study

Routine Bovine Viral Diarrhoea Virus (BVDV) monitoring of a commercial beef herd in southern New South Wales over a 10-year period provided an opportunity to assess the impact of the introduction of BVDV on that herd. BVDV antibody testing provided strong evidence that the herd was initially free of BVDV (2009–2011). Testing from 2012 suggested BVDV had been introduced into the herd and this was confirmed in 2015 with the identification of persistently infected (PI) animals. Having become established in the herd, the owners then set out to eliminate BVDV from the herd. Antigen testing aimed at identifying PI animals revealed BVDV was already absent from the herd. Subsequent antibody testing confirmed that the herd was now free from BVDV. Despite the incursion of BVDV in this herd, there was little measurable impact on reproductive performance (pregnancy rates), although suspected increased calf losses from birth to calf marking were reported. This is the first time such self-clearance has been documented as part of a longitudinal study under Australian conditions.     

Serological evidence for exposure to bovine viral diarrhoea virus in sheep co-grazed with beef cattle in New Zealand

ABSTRACT

Aims: To determine whether sheep that co-grazed with cattle that were suspected to be positive for bovine viral diarrhoea (BVD) virus had serological evidence of exposure to the virus.

Methods: Eighteen commercial farms that routinely co-grazed cattle and sheep in the same paddocks were recruited through purposive sampling. The recruiting veterinarians identified nine farms with cattle herds that were known or highly suspected to be positive for BVD and nine farms that were considered to be free of BVD. Blood samples were taken from 15 ewes aged 1 year on each farm and samples were submitted to a commercial diagnostic laboratory to test for antibodies against pestiviruses using an ELISA. All samples that were positive were then tested using a virus neutralisation test (VNT)for antibodies against BVD virus.

Results: Of the 270 blood samples, 17 were positive for pestivirus antibodies by ELISA and these originated from two farms that were known or suspected to have BVD virus-positive cattle. None of the samples from the nine flocks co-grazed with cattle herds that were known or suspected to be BVD virus-negative were positive for pestivirus antibodies. Within the two positive farms, 2/15 samples from the first farm and 15/15 samples from the second farm were antibody-positive. When the 17 positive blood samples were submitted for VNT, all 15 samples from the second farm tested positive for BVD virus antibodies with the highest titre being 1:512.

Conclusions and clinical relevance: In this small sample of New Zealand sheep and beef farms with suspected BVD infection in cattle, there was evidence of pestivirus exposure in co-grazed sheep. Although we were unable to confirm the origin of the exposure in these sheep, these findings highlight that farmers who are trying to eradicate BVD from their cattle should be mindful that the infection may also be circulating in sheep, and both populations should be considered a possible risk to each other for generating transient and persistent infections. Further work is needed to estimate the true prevalence of New Zealand sheep flocks that are affected by BVD and the associated economic impacts.