一株高病毒载量 PCV2 毒株的基因组特征及序列分析

【目的】了解广东省某猪群中猪圆环病毒 2 型（Porcine circovirus type 2,PCV2）流行毒株的遗传进化情况,丰富 PCV2 分子流行病学数据,为当地 PCV2 疫苗候选株的选用和研发提供参考。【方法】使用 qPCR 方法对疑似 PCV2 的样品进行检测,发现 1 株具有高病毒载量的 PCV2 毒株,命名为 GD222858。通过 PCR 方法进行全基因组分子克隆及遗传进化分析。使用 MegAlign 软件将该毒株 ORF1、ORF2 基因编码的氨基酸序列与 PCV2 同亚型参考毒株进行比对,分析氨基酸序列的相似性;采用 DNAStar 预测该毒株的 Cap 蛋白二级结构及 B 细胞表位,并与 4 株疫苗株 DBN-SX07-2（HM641752）、LG（HM038034）、SH（HM038027）、ZJ（AY686764）的 Cap 蛋白抗原指数进行比对分析。【结果】GD222858 毒株基因组长度为 1 767 bp。遗传进化分析表明该毒株属于 PCV2d 亚型。与国内外 82 株参考毒株的核苷酸相似性为 91.4%~99.6%,与越南毒株 Han8（GenBank 登录号：JQ181600）的亲缘关系最近。在 ORF1 编码的 Rep 蛋白处发现多个特异性突变位点 F70Y、F77L、W202R、N256S;ORF2 编码的Cap 蛋白相对保守。Protean 预测 Cap 蛋白的氨基酸第 5~18、24~25、39~41、48~49、57~65、99、101、112~114、139~140、145~150、162~165、175~181、188~189、205~211、227~232 位 置 处 均 可 能 存 在 潜 在 的 B 细 胞 表 位。GD222858 毒株的 Cap 蛋白抗原指数与 4 株疫苗株均有差异,在氨基酸 45~57、124~132、223~233 位置处抗原指数明显高于 4 株疫苗株,且与疫苗株 HM038034 差异最大。【结论】GD222858 毒株感染猪群的原因可能是 Rep 蛋白多个位点发生特异性突变及疫苗株选用不当所致。     

非洲猪瘟形势下猪流行性腹泻的防控措施

自2018年第一例非洲猪瘟疫情报道至今已经两年多了.为了快速扩繁猪群,许多清空场及新建场不得不大量外购来源复杂后备猪进群.因此,猪场在对猪流行性腹泻的防控上面临着更大的挑战.文章介绍了病毒性腹泻暴发的原因、防控难点和综合防控措施,期望为养殖业同人在实际生产中,科学防控猪流行性腹泻提供一定的参考借鉴.     

早期断奶仔猪营养性腹泻的分析及防治

仔猪腹泻是一种严重制约养猪业发展的疾病,其发生由应激、环境、营养等多种因素造成。本文主要从营养方面阐述了早期断奶仔猪发生腹泻的原因,并综合近年研究资料,论述了各种营养因素对仔猪营养性腹泻的改善作用,为研究合理的断奶仔猪日粮提供理论依据。     

鹿源牛病毒性腹泻病毒核酸疫苗免疫实验研究

试验应用间接ELISA方法检测鹿源牛病毒性腹泻分离株CCSYD核酸疫苗体液免疫效果,应用淋巴细胞转化实验检测其细胞免疫效果,并与鹿源牛病毒性腹泻分离株CCSYD灭活苗和标准株C24V灭活苗进行比较。结果表明:鹿源牛病毒性腹泻分离株CCSYD核酸疫苗产生了较好的体液和细胞免疫应答,免疫前后免疫应答水平差异均显著(P<0.05)。     

牛星状病毒EvaGreen实时荧光定量PCR检测方法的建立及应用

牛星状病毒(BAstV)是我国新发的犊牛腹泻病原,本试验的目的是建立检测BAstV的Real-time PCR方法.根据BAstV流行株的ORFla基因序列设计引物,通过优化反应条件和体系,成功建立基于EvaGreen检测BAstV的Real-time PCR方法.结果表明,该检测方法的Ct值与标准品模板在1.36×1...     

海南辣椒、樱桃番茄病毒病调查及病原dot-ELISA鉴定

[目的]调查和鉴定海南省辣椒和樱桃番茄病毒病发生情况和病毒种类,明确病毒病发生为害程度及毒源种类.[方法]2018—2019年,通过5点取样法对海南省10个市县的辣椒和樱桃番茄进行病毒症状类型、发病率和病情指数统计.采集辣椒和樱桃番茄主产区病毒病样本593份,通过斑点酶联免疫吸附法(dot-ELISA)对黄瓜花叶病毒(...     

Experimental transmission of encephalopathy and retinopathy induced by nodavirus to sea bream, Sparus aurata L., using different infection models

The transmission of viral encephalopathy and retinopathy (VER) was investigated in juvenile sea bream, Sparus aurata L. Two different infection routes [intraperitoneal (i.p.) and intramuscular injection (i.m.)] were tested at two different temperatures (20 and 26 °C) using sea bream of mean weight 0.7, 2 and 4 g, as well as an immersion challenge performed at 26 °C with sea bream of 0.7 g. Successful transmission of the disease was only achieved by i.m. injection. Mortalities of 100% occurred in sea bream of 0.7 g at day 15 post-infection and 47% in sea bream of both 2 and 4 g at day 30 post-infection in all the experimental infections at 26 °C. No mortalities were ever observed with infections at 20 °C. When mortalities were observed, the virus was detected by immunoperoxidase staining in the SSN-1 cell line inoculated with tissues from infected fish. Histological examination of both normal and infected fish showed a vacuolization in the bipolar and granular layers of the retina of the infected sea bream. This is the first experimentally induced transmission of VER in sea bream. Differences were observed at the time of disease onset depending on water temperature, the route of infection and the age of the juvenile fish.     

Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.     

Outbreak of viral haemorrhagic septicaemia (VHS) in lumpfish (Cyclopterus lumpus) in Iceland caused by VHS virus genotype IV

A novel viral haemorrhagic septicaemia virus (VHSV) of genotype IV was isolated from wild lumpfish (Cyclopterus lumpus), brought to a land‐based farm in Iceland, to serve as broodfish. Two groups of lumpfish juveniles, kept in tanks in the same facility, got infected. The virus isolated was identified as VHSV by ELISA and real‐time RT‐PCR. Phylogenetic analysis, based on the glycoprotein (G) gene sequences, may indicate a novel subgroup of VHSV genotype IV. In controlled laboratory exposure studies with this new isolate, there was 3% survival in the I.P. injection challenged group while there was 90% survival in the immersion group. VHSV was not re‐isolated from fish challenged by immersion. In a cohabitation trial, lumpfish infected I.P. (shedders) were placed in tanks with naïve lumpfish as well as naïve Atlantic salmon (Salmo salar L.). 10% of the lumpfish shedders and 43%–50% of the cohabiting lumpfish survived after 4 weeks. 80%–92% of the Atlantic salmon survived, but no viral RNA was detected by real‐time RT‐PCR nor VHSV was isolated from Atlantic salmon. This is the first isolation of a notifiable virus in Iceland and the first report of VHSV of genotype IV in European waters.