LRH─A3、hCG和PMSG对体外培养条件下牦牛垂体组织LH和FSH分泌的调节

促卵泡素３号（ＬＲＨ－Ａ３）、人绒毛膜促性腺激素（ｈＣＧ）和孕马血清促性腺激素（ＰＭＳＧ）等３种外源性激素均可增加体外培养条件下牦牛垂体组织分泌ＬＨ和ＦＳＨ的能力。培养液中ＬＨ和ＦＳＨ含量与加入的ＬＲＨ－Ａ３量呈正相关，与加入的ＰＭＳＧ和ｈＣＧ的量无显著关系     

2R,3R-丁二醇和2,3-丁二醇诱导匍匐翦股颖抗病性的比较

以2R,3R-丁二醇和2,3-丁二醇作为诱抗剂,在诱导匍匐翦股颖产生对褐斑病抗性的过程中,重点比较了不同施用方式的诱抗效果,筛选出了诱抗剂的最佳作用方式和浓度。结果表明:匍匐翦股颖接菌后第15d,2R,3R-丁二醇根部注射处理下,100μmol/L的病情指数最低,诱导效果最佳;而与叶面喷施相比,叶面喷施的诱导效果不明显;2,3-丁二醇根部注射处理下,250μmol/L的病情指数最低,诱导效果最佳,而与叶面喷施相比,叶面喷施的诱导效果不明显。结果表明,100μmol/L的2R,3R-丁二醇与250μmol/L的2,3-丁二醇根部注射可有效诱导匍匐翦股颖产生对褐斑病的抗性。     

硫酸铜,硝酸银和脱落酸对草地早熟禾胚性愈伤诱导的影响

试验以草地早熟禾4个品种的成熟种子为外植体,研究了CuSO4、AgNO3、脱落酸(ABA)对草地早熟禾胚性愈伤诱导的影响,结果表明,K3诱导培养基中添加CuSO4对草地早熟禾胚性愈伤组织的形成有促进作用,除DP192-10之外的3个品种都发生了明显的形态上的变化,DP-LF-300和2W42-117在添加5μmol/LCuSO4的培养基上诱导出了乳白色、致密的胚性愈伤组织,2W42-117的胚性愈伤率高达40%。在MS基本培养基中添加AgNO3、ABA对草地早熟禾4个品种的胚性愈伤组织的诱导没有作用。     

西安荷斯坦奶牛群5个基因座位遗传多态性的PCR-RFLP分析

应用PCR-RFLP方法对西安荷斯坦牛的κ-en、β-lg、β-lg5′侧翼区、CSN1S2、IGFBP-3共5个基因座位进行了多态性分析。结果表明，在西安荷斯坦牛群中，没有发现携带CSN1S2^P等位基因的个体，其多态信息含量为0。κ-en基因座位呈现低度多态(PIC=0．2366)，β-lg、β-lg5′侧翼区、IGFBP-3基因座位的多态信息含量分别为0．3168、0．3689、0．4439，均呈现中度多态。κ-en、β-lg、β-lg5′侧翼区、IGFBP-3、CSNIS2基因座位的杂合度和DNA多态度分别为0．2742、0．3947、0．4879、0．4891、0和0．0255、0．0116、0．0333、0．0112、0。而且，在西安荷斯坦牛群中，κ-en、β-lg、β-lg5′侧翼区、IGFBP-3共4个基因座位均处于Hardy-Weinberg平衡状态，CSN1S2基因座位处于纯合状态。     

亚洲璃眼蜱唾液腺一新功能基因的克隆与测序

HaB1是亚洲璃眼蜱雌成蜱唾液腺差异表达基因文库中的一个片段,根据其序列设计引物HaB1-GSP1和HaB1-GSP2,以唾液腺总RNA为模板,RACE法扩增获得HaB1的未知3′-末端。测定该末端序列,进行序列拼接,设计全长引物5-′CCAGTCCGAAGGAAGGGCG-3′和5-′TCTC-CGGGCACGTGAAGTGTC-3′。以cDNA第一链为模板,扩增获得基因全长。经核查表明,该基因为一新基因(登录号AY803896)。     

基因枪轰击不同剂量小鹅瘟病毒VP3基因疫苗在雏鹅体内的动态分布

本文开展了检测小鹅瘟病毒(GPV)VP3基因的实时荧光定量PCR(FQ-PCR)方法的建立和基因枪轰击不同剂量(1、3和6μg)GPV-VP3基因疫苗(pcDNA-GPV-VP3)在30日龄四川白鹅体内(心、肝、脾、肺、肾、法氏囊、胸腺、哈氏腺、十二指肠、空肠、回肠、直肠、盲肠、胰腺、血液、脑及注射部位皮肤)分布规律的研究。结果表明:①建立的FQ-PCR特异性强、灵敏度高、重复性好,核酸模板数与FQ-PCR测定的Ct值相关系数达到0.999,具有很好的直线相关性;②pcDNA-GPV-VP3各剂量免疫雏鹅1 h即可在各组织中检测到,其中注射部位含量最高,肝、肾、淋巴器官(脾、法氏囊、胸腺、哈氏腺)含量较高;③到免疫后217 d时,1μg组免疫雏鹅各个组织器官内仍检测到pcDNA-GPV-VP3的存在,但多数组织器官中的含量比1 h时约少了4个数量级,其中免疫部位减少了7个数量级;④血液中pcDNA-GPV-VP3的含量较少,且免疫后1 h~217 d各时间点的差异不显著(P≥0.05);⑤不同剂量pcDNA-GPV-VP3免疫雏鹅各组织中的含量呈现的总体规律为6μg组>3μg组>1μg组,但差异不显著(P≥0.05)。因此,FQ-PCR是定量检测pcDNA-GPV-VP3在免疫雏鹅体内含量的可靠方法,pcDNA-GPV-VP3免疫雏鹅后1 h时可分布至雏鹅体内各组织器官中并持续存在217 d以上。     

乳牛口服大量β-胡萝卜素对维生素D3及钙磷代谢的影响

６头健康成年荷斯坦乳牛被随机均分为２组。试验组牛第１周每头口服β－胡萝卜素２３．０ｇ,以后每周每头口服１１．５ｇ,对照组不做任何处理。试验期共８周。结果表明,试验组乳牛血清钙及无机磷水平呈下降趋势;血清维生素Ｄ３活性代谢产物２５－ＯＨ－Ｄ３和１,２５－（ＯＨ）２－Ｄ３均呈现明显下降;至第５周末,血液β－胡萝卜素水平最高,２５－ＯＨ－Ｄ３水平下降非常明显。据此认为,乳牛对β－胡萝卜素的吸收存在一阈值,大剂量的β－胡萝卜素进入机体,可能直接干扰维生素Ｄ３的羟化,使其活性代谢产物呈下降趋势,但由于吸收阈值的存在,其对维生素Ｄ３和骨骼钙磷代谢的干扰作用可得到一定程度的减缓。     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype

A new assay for the detection of swine influenza virus (SIV) was developed with a novel nucleic acid probe——Molecular beacon in this study. The specific primers and molecular beacon probes were designed according to the conserved region of H3 and N2 genes of SIV H3N2 subtype. A digital RT-PCR assay was developed for detection of SIV H3N2 subtype. The results showed that SIV H3N2 subtype could be identified simultaneously on this microarry with high sensitivity and reproducibility,which could reach to 10⁶ dilute viruse. The conclusion was that the digital RT-PCR method could analyze quantitatively the RNA templates.On the identification of H3N2 SIV,the digital RT-PCR method was much more scientific than Real-time quantitative PCR method.     

Molecular Characterization of Equine Rotavirus Group A Detected in Argentinean Foals During 2009–2014

Equine rotavirus group A (RVA) has been detected in several countries worldwide since its first detection in 1975. Currently, equine RVA is considered the major cause of dehydrating diarrhea in foals younger than 3 months, and the frequency of detection in clinical cases varies from 20% to 77%. The genotypes of epidemiologic relevance found in horses are G3P[12] and G14P[12]. In a survey conducted in Argentina from 1992 to 2008, equine RVA was detected in 21% and 39% of the fecal samples and outbreaks, respectively. Genotype distribution was 51% G3P[12] and 33% G14P[12]. In continuation with the surveillance, the aim of the present study was to characterize the equine RVA detected in Thoroughbred foals in Argentina from 2009 to 2014. A total of 436 stool samples (corresponding to 177 single diarrhea cases or outbreaks) were analyzed. Equine RVA was detected in 31% (135 of 436) of the samples, which corresponded to 42% (74 of 177) of outbreaks. From the positive cases, 42% (57 of 135) were genotyped. Of this, 63% were G3 (36 of 57) and 37% (21 of 57) were G14 genotype. Considering the whole data (1992–2014), equine RVA was detected in 25% (300 of 1,207) of the stool samples and 41% (119 of 293) of the diarrhea outbreaks. The results of this study also show a cyclic pattern of the G3 and G14 prevalence in the horse population with a change in G3:G14 frequencies from year to year. Furthermore, clustering in the phylogenetic tree suggests evolutionary and geographic relationships between the Argentinean strains compared with the strain circulating worldwide.