Study on Synergistic Effect of VA5 Immunopotentiator on Pigeon NDV Inactivated Vaccine

This study was designed to evaluate the synergistic effect of VA5 immunopotentiator on pigeon ND4416 inactivated vaccine.160 healthy pigeons were randomly divided into four groups,including ND4416 strain inactivated vaccine group (NDV group),ND4416 strain inactivated vaccine and immunopotentiator (VA5) mixed group (NDV+VA5 group),La Sota inactivated vaccine group (La Sota group) and normal saline as the control group (C group),to assess their vaccine efficacy against virulent pigeon NDV by serological analysis and animal testing.Pigeons sera were collected at different time points after immunization and measured the HI antibody titer of each group.The results showed that VA5 immunopotentiator significantly improved the serum antibody level of pigeons immunized with pigeon Newcastle disease inactivated vaccine (P<0.05).In addition,comparative test of spleen lymphocyte transformation was conducted at various time points after immunization.The results indicated that VA5 effectively stimulated the lymphocyte transformation of immune pigeon.Pigeons in each groups were challenged with ND4416 strain at the 30th,90th and 180th d after immunization.The results presented that the NDV+VA5 group had 100% protection rate and higher than La Sota group.The duration of immunization test showed that the antibody titer of NDV+VA5 group reached peak 11.20log₂ at the 21st d,remained 7.50log₂ at 180th d,and the protection rate remained 100% at 180th d.It indicated that VA5 immunopotentiator sustained the immune duration of pigeon NDV vaccine up to 180 d.Moreover,the in vitro detoxification test results suggested that VA5 immunopotentiator reduced the in vitro detoxification cycle of pigeons after challenge.Overall,this study suggested VA5 immunopotentiator could significantly improve the immune efficacy of pigeon Newcastle disease inactivated vaccine,which provided a basis for further enhancing the efficacy of Newcastle disease vaccine,as well increased experimental data for the application of immunopotentiators.     

柔嫩艾美耳球虫表面抗原SAG2基因的克隆与表达

根据生物信息学预测的基因序列设计引物,应用RT-PCR方法从柔嫩艾美耳球虫第二代裂殖子总RNA扩增获得了鸡柔嫩艾美耳球虫表面抗原2(surface antigen 2,SAG2)基因序列,将其与pGEM-T easy载体连接后转化E.coliDH5α,筛选阳性克隆,以带有限制酶切位点的特异性引物用PCR方法扩增不含SAG2 N端信号肽序列的ORF序列后克隆至表达载体pET-32 a(+),构建了重组表达质粒pET-32 a(+)-SAG2,并将其转化至E.coliBL21(DE3)。经IPTG诱导,获得了SAG2重组抗原在大肠杆菌的高效表达,重组蛋白的表达量约占菌体总蛋白的35%,融合蛋白的分子量约为47 ku。菌体经超声处理后进行SDS-PAGE分析表明,表达蛋白以包涵体的形式存在。     

不同施肥水平与组合对饲用黑麦经济效益的影响及施肥决策

应用回归均匀设计构造试验方案,研究氮肥、磷肥、钾肥不同水平与组合对饲用黑麦经济效益的影响及优化施肥方案。结果表明,饲用黑麦干草价格较低时,应减少肥料投入以降低生产成本为主,使所获利润最大;干草价格较高时,应适量增加肥料投入以提高产量,从而获得较高利润; 在饲用黑麦生产中,施肥量多少取决于所追求的目标。以产量为目标,就要投入较高的施肥量,以经济效益为目标,就要求适宜的施肥量,考虑到牧草品质,就要求较多的施氮水平。只有饲用黑麦产品实行优质优价,才能基本达到三者目标的统一。综合考虑,施肥量投入应为N:225 kg/hm²左右,不应低于150 kg/hm²,P₂O₅为75 kg/hm²,K₂O为30 kg/hm²以下比较适宜。     

草原红牛不同组织雄激素受体基因表达量检测及比较研究

采用实时荧光定量RT-PCR技术检测雄激素受体基因(Androgen receptor gene,AR)在草原红牛公牛和母牛多种组织中的表达情况,探讨该基因在草原红牛公牛和母牛不同组织中的表达差异.结果表明,AR基因在所检测的公牛和母牛的心、肝、脾、肺、肾、肠、胃及睾丸(卵巢)等8种组织中均有表达,且肝脏中的表达量高于其他组织;草原红牛公牛和母牛相同组织闻比较结果显示:AR基因在草原红牛公牛肝脏中表达量与母牛相比较差异不显著,其他不同组织间表达量均有显著差异(P<0.05),且公牛各组织中的表达量均高于母牛.本试验为深入研究AR基因的生物学功能及了解该基因对不同性别个体肉质性状的形成的作用机制奠定了基础.     

围产期奶牛代谢与营养调控研究进展

围产期是奶牛泌乳周期中最为关键的阶段,该阶段奶牛代谢的最大特点是能量摄入减少而需求增加从而导致能量负平衡.改善围产期奶牛的营养条件,最大限度地缓解能量负平衡,对于充分发挥奶牛的生产性能至关重要.本文综述了围产期奶牛的代谢特点以及营养调控研究进展,旨在为提高奶牛的生产性能和健康水平提供新的思路.     

Trinexapac-ethyl、Prohexad ione-Ca在草坪化学控制中的应用研究

为探明Trinexapac-ethyl和Prohexadione-Ca用于草坪化学控制的可行性,研究了其对高羊茅(Festuca arundinacea)生长发育的影响。结果显示:使用剂量高于0.095 kg/hm²时,Trinexapac-ethyl对高羊茅具有明显的矮化作用,在0.36kg/hm²剂量下Prohexadione-Ca的抑制率可达95.2%;Trinexapac-ethyl和Prohexadione-Ca都能使植株新增鲜重显著降低,但对新增干重影响很小,Trinexapac-ethyl对植株新增干重无影响;在施用Trinexapac-ethyl的情况下,随着留茬高度的增加新增株高减少,合理留茬高度为4-6cm;Trinexapac-ethyl和Prohexadione-Ca可以有效降低株高,从而减少草坪修剪次数。     

盐碱胁迫下黑麦草生长及离子微区分布特征

盐碱生境通过离子毒害和渗透胁迫等抑制植物的生长发育。离子微区分布调控主要通过调整盐分在不同部位的分布来减轻Na⁺等高浓度离子对重要组织器官的毒害作用,是盐碱胁迫下植物生存的重要策略之一。为探究黑麦草在盐碱胁迫下的离子微区分布特征,本研究通过水培法,采用不同盐碱浓度(0、50、100、200 mmol·L^-1)培养液处理黑麦草植株,考察其生长、离子吸收、运输选择性和微区分布变化,揭示黑麦草对盐碱胁迫的适应机制。结果表明:1) 盐碱胁迫下,植株的相对含水率、叶绿素含量逐渐降低,电解质外渗率显著升高;当盐碱浓度大于100 mmol·L^-1时,根长、株高和生物量均显著降低,即黑麦草的耐盐阈值是100~200 mmol·L^-1;2) 盐碱浓度为50 mmol·L^-1时,植株根中K⁺/Na⁺较空白升高了48.3%,Ca²⁺/Na⁺升高了54.1%,说明黑麦草根部可通过增强K⁺、Ca²⁺的吸收来维持细胞渗透压并缓解高浓度Na⁺的毒性;3) 在50~200 mmol·L^-1的盐碱浓度下,黑麦草叶片中可溶性组分及细胞壁中Na⁺的相对含量较对照显著增加,但细胞器内Na⁺的相对含量降低,说明叶部将Na⁺富集在液泡和细胞壁中,以减少Na⁺对细胞器的毒害作用;4) 叶片微观结构的SEM电镜图片显示,盐碱胁迫下叶片表皮增厚、导管数量减少和孔径缩小。综上所述,盐碱胁迫下黑麦草不同器官的适应机理不同,根部主要通过强化K⁺、Ca²⁺的吸收,降低对Na⁺的吸收,并将Na⁺区隔化在液泡中以降低离子毒害和维持渗透调节,而叶片主要通过将Na⁺区隔在液泡及细胞壁中,以及微观结构的变化来保护细胞器免受离子毒害。本研究可为黑麦草的耐盐碱适应机制及其在盐碱土上的种植提供理论依据。     

猪圆环病毒3型TaqMan-MGB荧光定量PCR方法的建立及应用

为建立猪圆环病毒3型(PCV3)荧光定量PCR检测方法,本研究根据GenBank中PCV3基因序列设计特异性引物和探针,经过反应体系和条件优化,建立了特异性检测PCV3的TaqMan-MGB荧光定量PCR方法。该检测方法在4.78×10~1拷贝/μL～4.78×10~9拷贝/μL质粒标准品范围内均有良好的线性关系;该方法特异性试验结果显示,其与多种常见猪病病毒均无交叉反应,特异性良好;本研究建立的方法敏感性是常规PCR方法的100倍,敏感性较高;批内批间重复性试验变异系数均小于2.3%,重复性良好。对临床样品的检测结果显示,该方法对PCV3的检出率高于常规PCR方法,并且PCV3阳性样品多存在混合感染情况。该方法的建立为PCV3的实验室诊断及流行病学调查提供了快速、准确的检测手段。     

Cloning,Expression and Bioinformatics Analysis of Enterococcus faecalis from Northeast Black Bear

In order to detect Enterococcus faecalis endocarditis antigen (EfaA) of bear that was used to disclose the infected Northeast Black bear immediately, a pair of PCR primers was synthesized by EfaA gene of Enterococcus faecalis that was recorded in GenBank (accession number:U03756.1) and the target fragment was 689 bp. The PCR product was cloned by pMD19-T vector, then was transferred to Escherichia coli DH5α competent cells and the positive clones were filtered. Recombinant plasmid (pMD19-T-EfaA) was extracted. Identification of PCR and digestion, sequencing, the structure prediction of proteins were operated. The results showed that EfaA gene was cloned successfully, and the gene homology with ATCC 29212 was 100.0%. pMD19-T-EfaA was constructed successfully, structure of proteins was predicted. This study provided theoretical basis for diagnostic methods of Enterococcus faecalis and laid basis for prevention and control of Northeast Black bear disease.