Development and application of G1-ELISA for detection of antibodies against bovine ephemeral fever virus

The gene encoding antigenic site G₁ of bovine ephemeral fever virus (BEFV) was highly expressed in the host cell Escherichia coli. An indirect G₁-ELISA with the recombinant protein as the coating antigen was established to detect antibodies against BEFV. The result revealed that the optimal concentration of the coated antigen was 0.5 μg/well and the dilution of serum was 1:20. It was optimal that sera with P/N value  2.2 were considered positive, P/N value  2.0 negative, and between 2.0 and 2.2 ambiguous. The G₁-ELISA method gave a sensitivity of 97.6% and a specificity of 98.6% by testing 590 field serum samples. These results suggest that the G₁-ELISA may be a good alternative tool for seroepidemiological surveys.     

Prime-boost immunization using alphavirus replicon and adenovirus vectored vaccines induces enhanced immune responses against classical swine fever virus in mice

This study was designed to evaluate the prime-boost vaccination regimens as a novel immunization strategy for DNA vaccine against classical swine fever virus (CSFV). BALB/c mice were primed with the alphavirus replicon-vectored DNA vaccine pSFV1CS-E2-UL49 encoding the E2 protein of CSFV fused with the UL49 gene encoding the transduction protein VP22 of pseudorabies virus, followed by either homologous boosting with pSFV1CS-E2-UL49 or heterologous boosting with the recombinant adenovirus rAdV-E2 expressing the E2 protein or with the baculovirus-produced recombinant E2 protein (rE2) in adjuvant. The humoral and cell-mediated immune responses following prime-boost vaccination were assessed. The results showed that: (1) boosting with either rAdV-E2 or rE2 elicited high-level antibodies, whereas homologous boosting with pSFV1CS-E2-UL49 elicited low-level antibodies (below positive threshold); (2) heterologous boosting with rAdV-E2 resulted in stronger CD8⁺ and CD4⁺ T cells proliferation responses and higher stimulation indexes; and (3) heterologous boosting with rAdV-E2 induced more IFN-γ production. These results support the notion that a regimen of DNA prime-recombinant adenovirus boost enhances humoral and cell-mediated immune responses, and the DNA prime-protein boost regimen enhances humoral immune responses.     

口蹄疫病毒受体猪源整联蛋白αvβ6的细胞定位及组织分布

利用pcDNA3.1（＋）-β6p真核表达载体稳定转染CHOK1细胞,间接免疫荧光法（IFA）检测整联蛋白αvβ6在转染细胞中的表达分布。以间接免疫组化法（IHA）检测整联蛋白αvβ6在健康猪舌、唇、气管等组织的表达分布。采用抗猪源整联蛋白β6亚基的单克隆抗体作为一抗,FITC标记山羊抗小鼠IgG、辣根过氧化物酶标记山羊抗小鼠IgG分别作为二抗。结果表明,pcDNA3.1（＋）-β6p真核表达载体稳定转染的CHOK1细胞的细胞膜及细胞质内可见致密的绿色荧光（IFA）,猪舌、唇、气管等组织细胞的细胞膜及细胞质内均出现了棕黄色的特异阳性反应物,表明整联蛋白αvβ6受体广泛分布于猪体的器官组织细胞中,为深入研究整联蛋白αvβ6在口蹄疫病毒感染过程中的作用奠定了基础。     

石羊河上游退化草地植物功能群生态位分异特征——以阿尔泰狗哇花型草地为例

采用时空转换方式,以石羊河上游高山草地阿尔泰狗哇花Heteropappus altaicus型退化草地及其原生群落为研究对象,以其主要功能群结构特征调查为基础,研究各功能群生态位季节动态特征。结果表明：该区生态位季节动态变化趋势同功能群类别有关,与功能群所处的演替阶段无关;功能群生态位数量特征变化的绝对数量及相对数量受到其所处的演替阶段影响,表现为随季节的推移禾本科功能群生态位宽度上升,其上升相对值为退化样地（120.8%）高于原生群落（65.5%）;而杂类草下降,其下降相对值差异较小,依次为51.38%和66.34%。随季节的推移禾本科-杂类草生态位重叠度升高,且升高的幅度为退化样地(113.3%)略低于原生群落(119.4%),就升高的绝对值而言,原生群落各观测时期均低于退化草地;杂类草-禾本科生态位重叠度则逐渐降低,其相对数量在退化样地与原生群落中依次下降53.0%和54.9%。其中退化草地禾本科、杂类草生态位重叠度在7月中旬存在一个拐点,其前期禾本科-杂类草组对低于杂类草-禾本科组对,而后期刚好相反。造成上述现象的原因可能与功能群生长特性和放牧干扰共同作用有关。返青较早的禾本科功能群早期生长受到放牧干扰的影响,为返青较迟的杂类草生长提供较大的资源空间,从而使得杂类草功能群在“植被空隙中进行季节更新”,进而改变了原群落的功能群结构。阿尔泰狗哇花型退化草地是植物群落退化演替过程中的关键阶段,如果加以适当的管理,草原退化将得到一定程度的改善;如果不加以保护,草地继续退化后的不可预知风险将提高。     

奶牛全混合日粮添加过瘤胃脂肪饲养效果对比试验研究

[目的]为了研究日粮中添加过瘤胃脂肪能够有效降低奶牛产后负平衡效果.[方法]本试验选用围产期(产前3～4周)健康的奶牛48头,按照年龄、体重、胎次、体况评分、预产期相同或相近的原则,参考上胎产奶量或母亲产奶量等指标随机分为试验组和对照组,在试验组日粮中添加过瘤胃脂肪300 g/头·日,试验期145 d.[结果]表明:试验组日产奶量提高1.96 kg,产后初次配种间隔缩短8.86 d,差异极显著(P＜0.01);对奶牛体况的改善也有一定的作用,特别是对头胎牛的体况改善明显.[结论]在奶牛全混合日粮中添加过瘤胃脂肪可提高奶牛产奶量、繁殖性能,改善体况.     

低磷日粮中添加植酸酶对蛋鸡肠道形态结构的影响

试验旨在研究在低磷的玉米-豆粕型日粮中添加植酸酶对蛋鸡肠道形态结构的影响。选用18周龄体重、产蛋周龄相近的伊莎褐蛋鸡840羽,随机分成7组:正、负对照组及在负对照组基础上分别添加300、1000和10000U/kg的包衣状植酸酶、300U/kg的粉状及300U/kg的微丸状植酸酶(试验Ⅲ、Ⅳ、Ⅴ、Ⅵ和Ⅶ组)。结果表明:与负对照组相比,Ⅳ组的空肠绒毛高度得到了显著提高(P<0.05),Ⅲ、Ⅴ、Ⅵ和Ⅶ组与之相比无显著差异(P>0.05),但分别提高了16.49%、21.80%、15.73%以及17.29%;隐窝深度、绒毛高度/隐窝深度、黏膜厚度与之相比差异不显著(P>0.05)。各试验组间以及试验组与正对照组之间亦无显著差异(P>0.05)。因此,在低磷日粮中添加植酸酶对蛋鸡肠道结构具有一定的改善作用。     

广西“猪高热综合征”主要病原调查及PRRSVNsp2基因变异分析

收集广西境内39个市(县)137个不同规模猪场及农村散养户146份病料,经检测发现广西"猪高热综合征"主要病原以PRRSV变异株(NSP2基因缺失)为主,检出率高达59.59%,其次为PCA占41.78%,CSFV占4041%,SIV占10.27%,PRV、HPS均占2.05%.将来自广西不同地区的15株PRRSV变异株(包括1株经典株)与国内外参考毒株部分Nsp2序列进行同源性比较,结果发现广西PRRSV变异株Nsp2序列之间同源性为83.3%～100%,与国内其他变异株同源性为83.2%～98.6%.这些毒株与国内其他变异株有完全一致的缺失特征.广西PRRSV变异株可分为2个亚群,亚群Ⅰ主要以桂西北地区毒株组成,亚群Ⅱ主要以桂东南地区毒株组成.所有的广西PRRSV流行株与国内其他毒株一样均属于美洲型.调查结果显示,广西"猪高热综合征"主要病原PRRSV变异株与PCV、CSFV等其他病原菌混合感染十分严重.其中,二重感染、三重感染分别高达42.53%和22.99%.     

犬瘟热RT-PCR快速诊断方法的建立

本研究建立了快速诊断犬瘟热感染的RT-PCR方法,并应用该方法对犬瘟热病死犬的脏器及感染犬体表采集样品进行了检测。为确定犬瘟热最佳的隔离检疫周期及采样部位,本研究对一例CDV攻毒犬感染病程收集的口鼻液、粪便、尿液样品进行了对比检测,结果证实粪便、口鼻液等易于获取的样品中CDV病毒滴度较高,并可在发病全程检测到,因此,它们可作为理想的受试样品。     

Effects of chromium stress on the subcellular distribution and chemical form of Ca,Mg, Fe,and Zn in two rice genotypes

A hydroponic experiment was carried out to study effects of chromium (Cr) stress on the subcellular distribution and chemical form of Ca, Mg, Fe, and Zn in two rice genotypes differing in Cr accumulation. The results showed that Ca, Mg, Fe, and Zn ions were mainly located in cell walls and vacuoles in roots. However, large amounts of metal ions were transferred from the vacuole to the nucleus and to other functional organelles in shoots. Chromium concentrations in the nutrient solution of 50 μM and above significantly decreased Ca concentrations in the chloroplast/trophoplast, the nucleus, and in mitochondria. It further increased Mg concentrations in the nucleus and in mitochondria, as well as Zn and Fe concentrations in the chloroplast/trophoplast. These Cr‐induced changes in ion concentrations were associated with a significant reduction in plant biomass. It is suggested that Cr stress interferes with the functions of mineral nutrients in rice plants, thus causing a serious inhibition of plant growth. The chemical forms of the four nutrients were determined by successive extraction. Except for Ca, which was mainly chelated with insoluble phosphate and oxalic acid, Mg, Zn, and Fe were extractable by 80% ethanol, d‐H₂O, and 1μM NaCl. The results indicated that these low–molecular weight compounds, such as organic acids and amino acids, may play an important role in deposition and translocation of Mg, Zn, and Fe in the xylem system of rice plants.     

三峡黄金旅游带旅游资源评价及开发研究

通过层次分析法、模糊赋分法对三峡工程蓄水后三峡黄金旅游带(重庆段)主要自然、人文景点(区)资源进行评价,并分析了旅游资源结构、开发价值及空间组合特点。在旅游带的旅游资源结构中,自然资源起着基础作用,人文景观为重要组成部分;在空间组合上可分为都市成片旅游区和沿江点状旅游区。基于上述评价结果,文章提出关于了三峡黄金旅游带旅游资源开发的一些思路。