Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Experimental investigation on the transport of different fish species in a jet fish pump

A jet fish pump with a throat of ø60 mm was designed to study its performance in the transport of different fish species and the physiological changes in fish thereafter. Experiments were conducted to investigate the fish transport rate and energy required to transport each ton of fish when transporting Carassius auratus, commonly known as the Chinese goldfish, Megalobrama amblycephala, or Wuchang bream, and Ctenopharyngodon idella, the grass carp. Fish were examined for external injuries as well as for several important enzymes and hormones which are indicators of tissue injury and stress. The results showed that the transport rate for all three species of fish rose dramatically with an increase in the primary stream rate. In this experiment, the transport rates of C. auratus, M. amblycephala and C. idella reached 2357 ± 37.2 kg ∙ h⁻¹, 2888 ± 41.6 kg ∙ h⁻¹, and 2060 ± 40.2 kg ∙ h⁻¹, respectively. However, both injury rate and energy required to transport each ton of fish increased no matter whether the primary stream rate was too low or too high. Considering both transport rate and injury rate, the mean primary stream rate of 80 m³ ∙ h⁻¹ was determined to be the optimal operating condition in this experiment. Fish were stressed and most likely some of their organs were damaged. However, most physiological indexes almost fully recovered after several hours.     

寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用

为建立一种针对寨卡病毒的快速诊断方法,本研究根据寨卡病毒的3’端保守基因序列,设计合成1对引物,建立了检测寨卡病毒的荧光定量PCR方法。结果显示:所建立的检测方法的Ct值与标准品在1.41×10^1~1.41×10^10^ copies/μL具有良好的线性关系,相关性为1,斜率为-3.502;灵敏性结果显示,该方法的检测限度为1.41×10^1 copies/μL,是普通PCR的10000倍;特异性结果显示,对CHIKV、DENV和JEV无特异性扩增,特异性强;重复性试验结果显示,组内和组间变异系数均小于1%,重复性好。本研究建立的SYBR Green I real-time PCR检测方法,可用于寨卡病毒感染的快速诊断。     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

宁南霉素等药剂对番茄黄化曲叶病毒病的田间防效

[目的]评价宁南霉素、艾德拉和碧护3种药剂混合使用防治番茄黄花曲叶病毒病的效果。[方法]通过田间试验研究8%宁南霉素、艾德拉绿(花、果)与碧护组成的药剂对番茄黄化曲叶病毒病的防治效果。[结果]8%宁南霉素、艾德拉绿(花、果)与碧护组成的试验药剂对番茄黄化曲叶病毒病具有很好的田间防治效果,第3次施药后10、20、30 d防效均在82%以上,与对照药剂番茄黄化曲叶病毒灵差异达极显著水平,且其产量比清水对照增加25 747.5 kg/hm~2,保产率为103.90%。[结论]宁南霉素、艾德拉和碧护混合使用对番茄黄花曲叶病毒病具有较好的防治效果,为番茄黄花曲叶病毒病的有效防治提供了理论依据。     

真核表达NDV F基因重组质粒的构建及其在CEF细胞中的转染

通过PCR方法自含NDVZF基因的克隆质粒中扩增NDVF基因 ,将其与真核表达载体pcDNA3..1/V5_His_TOPO重组。经核酸内切酶酶切 ,阳性克隆鉴定准确无误 ,核酸序列测定结果与原始基因比较 ,同源性大于 99% ,启始密码和终止密码未出现变异 ,将该重组质粒命名为pcDNANDVZF。将pcDNANDVZF在脂质体作用下转染CEF细胞 ,用间接免疫荧光试验检测 ,结果证明pcDNANDVZF可在CEF细胞中大量表达F蛋白。     

真核表达载体构建表达ＰＲＶ闽Ａ株gE基因表位抗原编码区片段的重组质粒

根据伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的序列与身份种真核表达载体pPICZaA、pAcGP67A序列与特性分别设计了两对ＰＣＲ引物。通过ＰＣＲ方法扩增到了两端具有不同酶切位点的gE基因表位抗原编码片段。将这２个片段分别克隆到pPICZaA与pAcGP67A载体，转化大肠杆菌ＴＯＰ１０菌档及ＸＬ１－Ｂlue菌株，获得了含伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的重组质粒pICZaA-FS与pAcGP67A-FS。序测定结果显示两个重组质粒中插入片段的大小与方向均正确。     

动物病毒性传染病的基本特征

动物病毒病几乎占传染病中的80％，经长期研究和查阅大量文献资料后，发现病毒性传染病具有许多独特的特征，了解掌握这些特征对病毒病的诊断，预防，治疗具有重要的指导作用。     

绵羊慢病毒自然感染绵羊的硬化性淋巴细胞性乳腺炎

７头来自新疆南部某绵羊慢病毒（ＯｖＬＶ）感染的羊场的绵羊用于本研究。用琼脂凝胶免疫扩散检查绵羊血清中对绵羊进行性肺炎（ＯＰＰ）病毒（ＯＰＰＶ）的抗体，结果表明有６例呈阳性，１例阴性，抗体效价在３年中呈下降趋势。４例血清学阳性边菜羊和１例阴性和田羊有不同程度的硬化性（纤维性）淋巴细胞性乳腺炎，小叶内有不等的淋巴细胞浸润，导管周围无淋巴滤泡形成，小叶间大量纤维组织增生。７例的肺、脑、关节、血管均无ＯｖＬＶ性特异性病变。从血清学阳性羊的外周血白细胞中未分离到ＯｖＬＶ。     

Transmission of the agent causing a melon yellowing disease by the greenhouse whitefly Trialeurodes vaporariorum in southeast Spain

The agent causing a yellowing disease of melon (Cucumis melo), which results in severe losses in crops under plastic on the coastal plains of southeast Spain, was shown to be transmitted in a semipersistent manner by the greenhouse whitefly (Trialeurodes vaporariorum Westwood). The agent was transmitted by grafting, but not by mechanical inoculation or through seeds. The agent was acquired in the minimum period tested (2 h) and could infect plants in an infection feeding interval of 6 h. Capsella bursa-pastoris, Cucumis melo, C. sativus, Cucurbita moschata, Cichorium endivia, Lactuca sativa andTaraxacum officinale were found susceptible.Results suggest that the yellowing disease affecting melon crops in the southeast of Spain is due to a pathogen similar to beet pseudo yellows virus, but this has to be confirmed by serology.