植物源杀菌剂大黄素甲醚对烟草病毒病的防治效果

为明确大黄素甲醚对烟草病毒病的防治效果,最佳施药时期及施用量。以0.1%大黄素甲醚水剂为试验药剂,8%宁南霉素水剂和20%盐酸吗啉胍可湿性粉剂为对照,开展田间小区试验。结果表明,0.1%大黄素甲醚水剂于移栽定植期施药防效最好,高浓度达62.81%,中、高浓度即制剂量80克/亩、100克/亩的防效与对照药剂均无显著性差异。田间观察发现大黄素甲醚对烟草有一定促生长作用。     

草莓对干旱胁迫生理及分子响应研究进展

干旱胁迫不仅会引起草莓大面积减产,还会直接影响果实的品质,已成为我国栽培品种草莓减产的主要非生物因子之一。本文主要综述干旱胁迫对草莓产量和品质、生理生化特性及部分基因表达的影响;此外,详细介绍了国内外在提高草莓干旱耐受性方面取得的研究进展和成果,以期为研究草莓耐旱机制和选育耐旱性优良的草莓新品种提供一定的参考依据和理论支持。     

河南省樱桃谷鸭乙型肝炎病毒全基因组克隆与序列分析

对河南省樱桃谷鸭主产区信阳、新郑、焦作三地628份樱桃谷鸭血清样本应用PCR技术进行鸭乙型肝炎病毒（DHBV）检测,并将三地DHBV阳性样本各挑选1份进行DHBV全基因的扩增、克隆、测序及序列分析。结果显示,信阳、新郑、焦作三地樱桃谷鸭DHBV自然携带率分别为10.5%、8.9%、17.6%;3株DHBV基因组全长分别为3 021、3 027、3 024bp,均含有编码P、S和C蛋白的3个开放阅读框,各开放阅读框氨基酸同源性比较显示差异显著的区域位于P蛋白。序列分析显示,3株DHBV河南株核苷酸同源性为89.8%~93.9%,与参考株为89.4%~99.5%。遗传进化分析及P蛋白关键位点分析结果表明,信阳分离株为西方基因型,其余2株为中国基因型。本试验掌握了河南省樱桃谷鸭主产区DHBV自然感染情况,并成功克隆了3株樱桃谷鸭DHBV全基因序列,为该病毒的进一步研究提供了有益信息。     

Two Akabane virus glycoprotein Gc domains induce neutralizing antibodies in mice

Akabane virus (AKAV), belonging to the genus Orthobunyavirus and family Peribunyaviridae, causes reproductive and congenital abnormalities in ruminants. Its envelope glycoprotein Gc is a neutralizing antigen, on which at least five distinct antigenic regions have been identified. We attempted to identify the domains using truncated recombinant AKAV Gc proteins expressed in Escherichia coli and monoclonal antibodies (mAbs) with AKAV-neutralizing activity. Dot blot analysis revealed that amino acid positions 1–97 and 189–397 (Gc_1–97 and Gc_189–397) in the truncated recombinant proteins reacted with the mAbs. Additionally, AKAV was neutralized by sera from mice immunized with these recombinant proteins. The results suggested that the two domains contain neutralizing epitopes and could be potential subunit vaccines against AKAV.     

猪脐静脉血管内皮细胞的分离及体外培养研究

为建立一种操作简单并且能获得大量猪血管内皮细胞的分离培养方法,本研究采集新生健康长白猪脐带,取其脐静脉血管,灌注0.1%的Ⅰ型胶原酶,于37℃水浴消化10 min后收集内皮细胞,培养于含20%胎牛血清的M199培养基中,长满单层后用胰酶消化传代培养。并对培养的细胞形态学、相关抗原和其他特征以及对病毒的易感性进行鉴定。结果表明,Ⅰ型胶原酶消化后可获得大量内皮细胞,细胞培养后的形态学观察显示细胞贴壁生长良好,呈典型的铺路石状排列;第Ⅷ因子相关抗原和细胞摄取乙酰化低密度脂蛋白均呈阳性,内皮细胞比率可高达100%。传3代后的生长曲线显示细胞传代后延迟期小于1 d,对数生长期约3 d,第5 d进入平台期,第7 d开始脱落死亡。病毒感染试验表明所获得的内皮细胞对猪瘟病毒高度易感,病毒生长与常用的PK-15没有区别。上述研究结果表明本研究建立的血管内皮细胞分离培养方法简便有效,为大量制备原代血管内皮细胞提供了可靠的方法。     

禽白血病病毒J亚群诱导蛋鸡群多种肿瘤

2009年8月,山东泰安某海兰褐商品蛋鸡场,开产后产蛋率只有60％～70％,31周龄时,死亡率突然增加,最高达20％.患病鸡经大体剖检、病理组织学、PCR和免疫组织化学检测确诊为J亚群禽白血病.病理组织学检测发现送检鸡除了髓细胞瘤外,还存在明显的纤维肉瘤、组织肉瘤、腺癌和血管内皮瘤等肿瘤.纤维肉瘤单独存在,组织肉瘤、腺癌和血管内皮瘤分别和髓细胞瘤混合存在,本文详细描述了各肿瘤的组织学特点.ALV-J从肉种鸡传向商品蛋鸡后引起肿瘤谱扩展的原因和机制值得进一步研究.     

犬瘟热病毒N蛋白基因融合蛋白原核表达条件的优化与蛋白纯化

[目的]提高犬瘟热病毒N蛋白基因在大肠杆菌中表达可溶性GST融合蛋白的含量.[方法]研究诱导剂(IPTG)的最佳加入时间、诱导剂(IPTG)浓度、诱导时间以及不同温度条件对GST - CDV N融合蛋白表达的影响,获得在大肠杆菌中的最佳的表达条件,然后再在最佳条件下大量的表达,用GST - Sepharose 4B亲和层析柱对GST - CDV N融合蛋白进行纯化,得到最大量的可溶性融合蛋白,并对蛋白含量进行测定.[结果]大肠杆菌BL21 (DE3)转化菌在37℃培养3.5 h(OD600 =1.314)时,加入IPTG使终浓度为1 mmol/L,然后在27℃诱导8h,GST - CDV N融合蛋白可溶性表达量最高,达到0.862 mg/mL.[结论]确定了犬瘟热病毒N蛋白基因的优化表达条件,为犬瘟热病毒分子免疫学诊断和亚单位疫苗的研制奠定了基础.     

Substitution of vaccinia virus Elstree by modified vaccinia virus Ankara to test the virucidal efficacy of chemical disinfectants

After the eradication of variola in 1980, the smallpox vaccination was considered to be no longer required and was subsequently abandoned mainly because of possible adverse effects of vaccinia virus especially in first‐time vaccinees. Despite a growing number of humans without immunity against vaccinia virus, vaccinia virus Lister Elstree (VACV) is still prescribed for testing virucidal efficacy of chemical disinfectants in the guidelines of the German Veterinary Medical Society [Deutsche Veterinärmedizinische Gesellschaft (DVG)], the German Association for the Control of Virus Diseases [Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten (DVV)] and the Robert Koch Institute (RKI). To evaluate a possible substitution of VACV, with the attenuated modified vaccinia virus Ankara (MVA) the virucidal efficacy of four different DVG‐listed commercially available chemical disinfectants representing different groups of chemicals was tested against these two viruses. Quantitative suspension tests and qualitative carrier tests with poplar wood and gauze were performed. Distinction of VACV and MVA was confirmed by cytopathogenic effects, such as differences in plaque morphology. No significant difference in disinfection efficacy between VACV and MVA was observed for any of the disinfectants tested. Implying that vaccinia virus poses a risk after inadvertent inoculation, our results show that MVA, which does not replicate in humans, should replace VACV in the chemical disinfectant testing guidelines.