Characterization of polymorphisms of transferrin receptor and their association with susceptibility to ETEC F4ab/ac in pigs

Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) expressing F4 (F4ab, F4ac and F4ad) fimbriae is a significant cause of mortality and morbidity in newborn and weaned pigs. The locus controlling susceptibility towards ETEC F4ab/ac has been mapped to SSC13q41, in which TFRC (transferrin receptor) was localized and considered as a positional candidate gene for ETEC F4ab/ac receptor. In this study, we determined susceptibility/resistance to ETEC F4ab/ac in a total of 755 F2 animals from a White Duroc x Erhualian intercross using a microscopic enterocyte adhesion assay. We identified two TFRC polymorphisms (SNPs 591 A>G and 632 A>G) in a single exon after comparative sequencing analysis of 2371-bp amplicons containing the complete coding region of TFRC using RNA of eight full-sib F2 animals with susceptible and resistant phenotypes. The intron sequences flanking the two exon polymorphisms were obtained, revealing an intron polymorphism (SNP 291 C>T). We genotyped the 19 founder animals of the White Duroc x Erhualian intercross for the identified polymorphisms, showing that only the 291 C>T polymorphism is a highly informative marker. We further genotyped all 59 F1 and 755 F2 animals for the 291 C>T polymorphism, and the association of this polymorphism with susceptibility/resistance to ETEC F4ab/ac in these F2 animals was evaluated by the transmission disequilibrium test. The result showed that the 291 C>T polymorphism is not a causal mutation, however, has a significant linkage disequilibrium with the ETEC F4ab/ac, especially F4ac receptor locus.     

工程菌DH5a感受态细胞的制备及质粒pGEM的转化研究

绿色荧光蛋白（green fluorescent protein，GFP）是源于海洋生物多管水母(aequoia victoria)的一种发光蛋白，能够自身催化形成生色团并在蓝光激发下发出绿色荧光，能在活细胞内稳定表达，本试验首先从含有GFP的大肠杆菌中提取质粒，并利用胶回收试剂盒纯化质粒，采用氯化钙法将工程菌DH_5a制成感受态细胞，然后将纯化后的质粒转入已制备好的感受态细胞中，使得工程菌DH_5a转化成具有抗氨苄青霉素的大肠杆菌，并表达绿色荧光蛋白，接种于加氨苄青霉素的LB培养基上，筛选阳性克隆。     

CpG-ODN enhances mammary gland defense during mastitis induced by Escherichia coli infection in goats

Seven healthy native goats in early lactation, weighing 30–40 kg, were used in this study. The right mammary gland of the seven does were infused with CpG-ODN at a dosage of 100 μg kg⁻¹ body weight on the day 5 postpartum (PP). The left glands were used as controls and infused with sterile phosphate-buffered saline (PBS). On day 8 PP, the same dosage of CpG-ODN or PBS was again infused. On day 9 PP, the mammary glands (both right and left) of the seven does were infused with 6 × 10⁶ colony-forming units (CFU) Escherichia coli and, at 0, 8, 16, 24, 48 and 72 h postinfection (PI), milk samples were collected from all glands. Goats were euthanized at 72 h PI and the mammary tissue harvested. Infusion with 6 × 10⁶ CFU ml⁻¹ E. coli induced acute mastitis. Histopathological evaluations showed that polymorphonuclear neutrophils (PMNs) were still present in alveoli at 72 h PI, but PMNs in the CpG-ODN-treated glands has disappeared. Bacteria counts in milk peaked at 16 h PI and CpG-ODN induced a significant decrease in viable bacteria from 16 h PI until the end of the experiment. This study showed that CpG-ODN promoted the expression of its specific receptor (TLR-9 mRNA) in mammary tissue, stimulated IL-6 production, reduced bacteria counts in milk, attenuated the impact of inflammation mediators on cells and significantly shortened the inflammation course. These results suggest that the CpG-ODN improved mammary gland defense and, thereby, had a beneficial effects against mastitis caused by E. coli infection in goats.     

2015年非洲口蹄疫流行状况与分析

2015年,非洲共有29个国家向世界动物卫生组织（OIE）通报发生771起口蹄疫疫情,包括A、O、SAT 1、SAT 2和SAT 3等5种血清型。O型和SAT 2型是非洲地区的主要流行血清型;病毒多通过野生动物散播;东非是非洲的主要疫区;感染家畜以牛为主。2015年非洲的口蹄疫流行状况和特点提示：该地区的口蹄疫防控任务十分艰巨,短期难以控制或根除;南非型口蹄疫病毒通过野生动物迁徙或非法动物产品贸易等途径向外扩散的风险依然存在,需提高警惕。     

12个多年生牧草品种在迪庆高寒地区的引种评价

为筛选出适宜云南迪庆高寒地区种植的多年生牧草品种,本研究开展了12个多年生牧草品种引种适应性的比较,并采用灰色关联度综合评价法对其生产性能进行了综合评价。结果表明,12种牧草生产性能的高低顺序为紫花苜蓿牧歌401+Z(Medicago sativa cv.Amerigraze401+Z)菊苣将军(Cichorium intybus cv.Commander)鸭茅安巴(Dactylis glomerata cv.Anmba)多年生黑麦草雅晴(Lolium perenne cv.Yatsyn)白三叶铺地(Trifolium repens cv.Prop)红三叶(T.pratense)白三叶海法(T.repens cv.Haifa)紫花苜蓿游客(Medicago sativacv.Eureka)紫花苜蓿WL-525(Medicago sativacv.WL-525)扁穗雀麦草地麦特(Bromus catharticus cv.Grasslands Matua)紫花苜蓿猎人河(M.sativacv.Hunterriver)老芒麦同德(Elymus sibiricus cv.Tongde)。其中,牧歌401+Z紫花苜蓿、将军菊苣、安巴鸭茅、雅晴多年生黑麦草、铺地白三叶、普通红三叶、海法白三叶在当地适应性较好,生产性能相对较高,可作为当地建植栽培草地及改良天然草地的优选品种。研究结果对解决当地饲草匮乏、促进当地草地畜牧业的可持续发展具有重要的意义。     

中国南方荷斯坦奶牛κ-CN基因第四、第五外显子的多态性研究

本文利用PCR-RFLP技术，检测了中国南方荷斯坦牛κ-酪蛋白（κ-casein，κ-CN）基因第4、第5外显子的遗传多态性。结果表明：κ-CN基因第4、第5外显子均存在遗传多态性，第4外显子上存在突变位点g．10943A〉C，第5外显子上存在突变位点g．15153T〉C。第4外显子有两种基因型（AA，AB），等位基因频率分别为0．9379和0．0621，第5外显子有三种基因型（AA，BB，AB），等位基因频率分别为0．6512和0．3488。κ-CN两个外显子PCR-RFLP位点的基因型分布符合Hardy-Weinberg平衡定律（p〉0．05）。κ-CN基因第4、第5外显子座位的多态信息含量分别为0．1097和0．3511，第4外显子呈现低度多态，第5外显子呈现中度多态；其杂合度、有效等位基因数分别为0．1165、1．1318和0．4543、1．8324。     

湖北省某县羊布鲁氏菌病群体血清流行率横断面研究

2015年初,湖北省某县启动羊布鲁氏菌病监测净化工作。为了解该县羊布鲁氏菌病的流行现状和阳性场户的地理分布,对该县1 567户养羊场户分两阶段抽样,采用虎红平板凝集和c ELISA垂直试验进行检测,获知该县羊布鲁氏菌病的群体血清流行率。结果显示:该县羊布鲁氏菌病真实群体血清流行率为2.00%,不同乡镇和不同养殖规模养羊场户的群体血清流行率分别在0~11.16%和0.67%~25.58%之间不等;检出布鲁氏菌病阳性场户74户,其分布在全县10个乡镇,主要集中在4个乡镇,以存栏量在50只以下的养羊场户居多;不同乡镇和不同养殖规模养羊场户中的布鲁氏菌病分布情况不同。调查结果提示,应根据本底调查结果,制订净化方案,实行布鲁氏菌病区域化管理,同时加强对不同流行区域间羊群移动的检疫监管。     

出生前山羊大脑皮质的形成和发育

对山羊胚胎大脑皮质形成及其神经元发育的形态学变化进行了研究，结果表明：（1）山羊胚胎不同类型大脑皮质的皮质板形成时间顺序为旧皮质、新皮质→海马皮质，与种系发生规律相反，提示神经系统内部结构的形成并非完全符合种系进化，而与动物个体组织结构自身的发展相关。不同类型大脑皮质皮质板的分化顺序为古皮质→旧皮质→新皮质，与种系发生的顺序相符。（2）山羊胚胎海马皮质锥体细胞的发育较早，按由深到浅的次序发育，而海马皮质中的非锥体细胞于第19周开始发育。梨状叶皮质锥体细胞发育的较早，最先出现在深层，颗粒层第18周出现。大脑新皮质各层神经元发育顺序依次为：内锥体层→外锥体层、多形层→内颗粒层→外颗粒层。各种类型皮质的神经元在出生前都未发育成熟。     

猪囊尾蚴囊液抗原制备及ELISA检测条件的优化

本试验采用SephadexG-200层析技术纯化猪囊尾蚴（Cysticercus cellulosae）的囊液粗抗原。共获得两种（CF1、CF2）层析抗原，以酶联免疫吸附试验（ELISA）判定抗原的最适包被浓度以及抗体的最适稀释倍数，各阶段最适反应条件，结果证明CF1具有特异性强、敏感性高、稳定性好、重复性强的特点可以作为免疫诊断囊尾蚴病猪IgG。     

两种早熟禾超氧物歧化酶活性测定的适宜条件

以冷地早熟禾 (Poacrymophila)和草地早熟禾 (P .pratensis)为材料 ,探讨光照反应时间、酶浓度以及酶反应体系等对NBT光化还原反应的影响。随着光照反应时间的延长 ,最大光还原值直线上升。 2 5min后光化还原值逐渐趋于稳定。光照反应开始后 ,酶液对NBT光化还原反应速率的抑制百分率迅速升高 ,在 15～ 2 5min时 ,酶液对NBT光化还原反应的抑制百分率保持稳定 ,2 5min后下降速率较大。光照反应时间对SOD酶活性测定的影响较大 ,确定为 2 5min较为适宜。酶浓度在 4 0～ 6 0mg/mL时光化还原值变化速率较一致。测定中反应体系NBT、核黄素的加样量以及光照强度在一定范围内时光化还原值较为稳定