水貂阿留申细小病毒的致病机制、鉴定及防治方法

<正>水貂阿留申细小病毒(Aleutian mink disease parvo virus,AMDV)是感染水貂的自主复制型细小病毒,是引起水貂阿留申病的病原。AMDV感染水貂后会产生抗体依赖的病毒感染增强作用(Antibody-dependent enhancement,ADE),还没有效果较好的疫苗进行防控。本文主要     

水貂阿留申病的研究进展

水貂阿留申病(Aleutian disease of mink,ADM)是由水貂阿留申病细小病毒(Aleutian mink disease parvovirus,AD-MV)引起的一种慢性、进行性传染病,一直是危害世界养貂业健康发展最重要的疫病之一。到目前为止,还没有疫苗可成功用于ADM的预防,也没有特异有效的治疗方法,唯一可行的防治方法就是通过多次特异性检疫,淘汰病貂,净化貂群。笔者对阿留申病的病原学、发病机制、防治措施等方面进行概述,为临床防治水貂阿留申病提供了理论基础。     

水貂阿留申病粪便中DNA的提取方法及与产仔数的相关性

<正>水貂阿留申病(Aleutian disease of mink),又称浆细胞增多症(Plasmacytosis),是由细小病毒科的阿留申病病毒(ADMV)引起的一种慢性传染病,导致自身免疫系统紊乱并逐渐衰竭,并发强烈的自身免疫。主要侵害网状内皮系统,其特征为浆细胞弥漫性增生、产生多量C-球蛋白和持续性的病毒血症。水貂阿留申病具有自身免疫     

水貂阿留申病的危害及净化措施

正水貂阿留申病(Aleutian disease of mink,AD)是一种以水貂繁殖能力下降、冬季饮欲增强、食欲减退、逐渐消瘦死亡为主要临床症状的病毒性传染病,而且本病严重损害水貂的免疫系统,造成疫苗的免疫保护期缩短、抗体整齐度参差不齐,严重影响疫苗的免疫效果。本文针对国内水貂养殖及防控现状,提供了合理的检测净化方法以供广大养殖户参考,为国内种貂选育和商品貂的繁殖工作奠定基础。水貂阿留申病(ADM)又称浆细胞增多症(plas-     

我国水貂阿留申病的检测及净化

水貂阿留申病(Aleutian disease of mink,AD),是以水貂繁殖能力下降、消痩、自身免疫病、高免疫球蛋白血症、肾衰竭死亡为基本特征的一种病毒性传染病。目前该病在我国呈现较高的感染率(60%～70%),严重影响我国养貂业的健康可持续发展。文中针对我国水貂阿留申病的流行及危害现状,提出合理的检测方法,为提升并稳定阳性貂的生产性能,逐步建立国内优良水貂育种群提供参考。     

水貂阿留申病毒感染过程中主要分子蛋白作用机理研究进展

正水貂阿留申病(Aleutian mink disease, AMD)是由AMD病毒(AMDV)感染水貂引起的自身免疫系统功能紊乱,并引发自身免疫的慢性、传染性疾病,在全球范围的水貂养殖国家造成了重大的经济损失~([1-2])。由于其特殊的致病机制和抗体依赖性增强作用(ADE)~([3-4]),尚无预防该病的商业疫苗。目前主要通过检疫淘汰病貂来净化貂厂,但     

水貂阿留申病的诊治

阿留申病(Aleutian disease of mink)是水貂的一种慢性病毒性传染病,主要特征是全身性浆细胞增多,血清丙种球蛋白增高,肾小球肾炎以及纤维素性坏死性动脉炎,持续性病毒血症和母貂空怀显著增加。本病最初在"阿留申"品系     

水貂阿留申病毒结构蛋白与非结构蛋白的研究进展

水貂阿留申病毒(Aleutian mink disease virus,ADV)是一种主要侵染水貂的自主复制型细小病毒,是一种在水貂中广泛存在的重要病原体。病毒粒子的蛋白分为结构蛋白(VP1、VP2)和非结构蛋白(NS1、NS2)两类。VP1蛋白对病毒粒子产生感染性有重要作用;VP2蛋白是主要免疫功能区,能刺激机体产生中和抗体;NS1和NS2主要参与病毒的复制和基因的表达调节。文中对近年来国内外学者关于水貂阿留申病毒结构蛋白和非结构蛋白的研究情况进行归纳和总结。     

水貂阿留申病毒荧光定量PCR检测方法的建立及初步应用

根据水貂阿留申病毒（Aleutian mink disease virus,AMDV）基因序列（GenBank登录号：NC_001662.1）设计1对特异性引物,通过对荧光定量PCR反应条件的优化,建立了检测AMDV的SYBR GreenⅠ荧光定量PCR方法,该方法的灵敏度达10拷贝/μL,≥10²拷贝/μL具有良好的特异性和重复性。同时利用该方法对8份疑似AMDV血清进行检测,结果6份阳性,阳性率为75%。本研究为水貂阿留申病的鉴别诊断及净群根除奠定了技术基础。     

水貂阿留申病基因工程诊断抗原的制备

本研究旨在建立一种生产水貂阿留申病(Aleutian disease,AD)诊断抗原的新方法。试验提取水貂阿留申病毒(Aleutian disease virus,ADV)的基因组,PCR扩增ADV核衣壳蛋白VP2基因,构建重组表达质粒Bacmid-VP2,脂质体介导其转染昆虫细胞Sf9获得重组杆状病毒AcMVPV-VP2。电镜下观察表达的VP2蛋白,Western blotting检测目的蛋白的反应原性。以传统接毒方法生产的AD诊断抗原作对照,通过对流免疫电泳试验检测表达蛋白的生物学活性。结果表明,表达的重组VP2蛋白在电镜下组装成病毒样颗粒且能与ADV阳性血清发生反应。与商业诊断抗原相比,重组抗原诊断AD的阴阳性的符合率为100%。该方法可成为生产AD诊断抗原的替代方法。     

Nucleotide sequence and polymerase chain reaction/restriction fragment length polymorphism analyses of Aleutian disease virus in ferrets in Japan.

Two ferrets with spontaneous Aleutian disease (AD) were found in Japan. The diagnosis was verified by polymerase chain reaction (PCR) amplification of part of the capsid gene specific to AD virus (ADV). The nucleotide sequences (365 bp in length) of the amplified fragments from the 2 ferrets differed by a single nucleotide, producing an amino acid alteration. Compared with other types of ADV, these isolates had 96% sequence similarity to a published ferret ADV (FADV) in contrast to <91% homology to various types of mink ADV (MADV). The phylogenetic tree of ADVs indicates that these 2 isolates and the published FADV belong to the same genetic group and definitely are divergent from MADVs. The predicted amino acid sequence of the hypervariable segment in the capsid gene was conserved among the 3 types of FADV. These results indicated that the 2 isolates found in Japan were new DNA types of FADV and could have been derived from FADV(s). A restriction fragment length polymorphism (RFLP) method to distinguish the ferret types of ADV from the mink types of ADV was developed on the basis of differences in their nucleotide sequences. Digestion of the PCR products with Afal or ScaI provided different cleavage patterns for FADV and MADV. This PCR/RFLP analysis of the ADV capsid gene will be a valuable asset for diagnosis of this virus infection in ferrets.     

水貂阿留申病发病机理的研究进展

水貂阿留申病是由阿留申病病毒引起的一种可对水貂养殖业造成严重损失的传染病。该病困扰动物医学界多年 ,始终未得到攻克。近年来随着分子生物学技术的发展 ,国内外学者对该病分子水平的发病机理有了更多的认识 ,研究内容主要集中在与水貂阿留申病发病密切相关的抗病毒抗体、病毒核酸的晚启动子P3 6及其顺式作用元件(cis acting)、结构蛋白VP2等方面。文章对以上研究进展进行了详细的归纳总结与分析 ,并在此基础上提出了尝试使用反义RNA或干扰RNA对该病进行预防、治疗的设想 ,以期今后对阿留申病的继续深入研究能有所帮助。     

阿留申病毒强致病性毒株非结构蛋白基因的克隆及序列分析

根据Genbank公布的全基因序列设计两对引物,利用PCR扩增技术,得到阿留申病毒非结构蛋白基因ADV-LN1、ADV-LN2,将其克隆至pMD18-T载体中进行序列测定,并与其它国内外毒株的非结构蛋白进行序列比较和分析。序列分析表明,同国外毒株相比核苷酸序列同源率NS1为87.8%～99.1%,NS2为87.7%～98.5%,NS3为85.4%～99.2%;氨基酸序列同源率NS1为82.5%～98.0%,NS2为81.6%～96.5%,NS3为78.2%～98.9%;与国内毒株相比核苷酸序列同源率NS1为91.5%～93.8%,NS2为93.0%～94.7%,NS3为93.5%～96.6%;氨基酸序列同源率NS1为87.4%～89.9%,NS2为87.7%～90.4%,NS3为88.5%～94.3%。系统进化树分析表明:国内流行毒株之间与欧美毒株之间,差异逐渐缩小,遗传变异趋势更加复杂。     

Isolation,Identification and Drug Sensitivity Analysis of Shigella from Minks

In order to provide therapeutical guidance for drug admistration, the bacteria of three sick minks suffering from typical diarrhea symptoms provided by mink farms in Jilin province were isolated and identified, and the drug sensitivity was tested. The bacteria were isolated with TSA plates, and identified using biochemical methods and PCR assay. The virulence of the isolates was determined by infecting BALB/c mice. The antimicrobial susceptibility of the isolates to antimicrobial agents was investigated using the K-B method. PCR was used to detect the resistance genes and Ⅰ integrons. A total of 3 Shigella isolates were obtained from sick minks. The virulent determination showed that all isolates could cause mice diarrhea. The drug sensitivity results showed that 3 strains were sensitive to fluoroquinolone, cephalosporin, florfenicol and polymyxin, but they were resistant to aminoglycoside, tetracycline, chloramphenicol, penicillin and ampicillin. There were seven resistance genes were detected,bla_TEM-1,aadA1, aac(3')-Ⅱc, aac(6')-Ⅰb, aph(3')-Ⅶ, tet(M), cat2 and three class Ⅰ integrons carrying aadA 1 gene cassette. All of the isolates were virulent and caused the mice diarrhea. The resistance of the 3 strains were very serious and mainly for multiple drug resistance phenomenon. The resistance genes detected in the mink were various, and could bring enormous implications for clinical treatment.     

Isolation,Identification and Drug Sensitivity Analysis of Pseudomonas aeruginosa from Minks

In order to provide therapeutical guidance for drug admistration,the bacteria from five dead minks suffering with typical symptoms provided by mink farms in Shandong province were isolated and identified,and the drug sensitivity was tested.The bacteria were isolated with TSA plates,and identified using biochemical methods and PCR assay.The drug sensitivity of the isolates to antimicrobial agents was investigated using K-B method.PCR was used to detect the resistance genes.A total of 5 Pseudomonas aeruginosa isolates were obtained from sick minks.The drug sensitivity results showed that 5 strains were sensitive to fluoroquinolone,the third and fourth generations cephalosporin drugs,but were resistant to aminoglycoside,tetracycline,chloramphenicol,penicillin,the first and second generations cephalosporin drugs.There were six resistance genes were detected,aadA1,aac(3')-Ⅱc,aac(6')-Ⅰb,tetA,tetK and cat2.All of the isolates were detected more than three kinds of resistance genes.The result showed that 5 strains were all Pseudomonas aeruginosa,and Pseudomonas aeruginosa was the main cause of mink hemorrhagic pneumonia.The resistance of 5 strains were very serious and mainly for multiple drug resistance phenomenon.The resistance genes detected in the mink were various,and could cause the resistance to the drugs.     

水貂阿留申病毒地方强毒株ADV-LN的分离鉴定

本研究采用对流免疫电泳方法(CIEP)检测辽宁水貂养殖场疑似水貂阿留申病毒(aleutian mink disease virus,ADV)水貂血清抗体,采集抗体阳性水貂的肝脏、脾脏、肾脏和肠系膜淋巴结组织,电镜观察存在细小病毒样颗粒。组织液研磨无菌处理后,接种CRFK细胞,盲传6代,取病毒细胞分离液用PCR方法检测,呈ADV阳性。将病毒分离液纯化后接种健康水貂,隔离观察,接种后3 d即出现食欲减退,贫血,被毛无光泽,后期出现拒食、狂饮、死亡,个别水貂出现神经症状,表现抽搐、痉挛、步态蹒跚、共济失调,证明分离获得的病毒为ADV强毒株,命名为ADV-LN株。     

水貂绿脓杆菌分离鉴定及药物敏感性分析

本试验旨在通过对山东某水貂养殖场送检的5只具有典型出血性肺炎症状的病死水貂进行细菌分离鉴定及耐药情况分析,为临床用药和治疗提供依据和参考。通过细菌分离纯化、生化鉴定和PCR方法对分离菌株进行鉴定,采用K-B药敏法检测分离菌株对常用药物的敏感性,并通过PCR方法检测分离菌株耐药基因的携带情况。经鉴定共分离得到5株绿脓杆菌,药物敏感性试验结果显示,5株绿脓杆菌对氟喹诺酮类药物、第3代和第4代头孢类药物较敏感,对氨基糖苷类、四环素、氯霉素、青霉素类、第1代和第2代头孢类药物耐药。耐药基因检测结果显示,5株绿脓杆菌共检测出6种耐药基因aadA1、aac(3')-Ⅱc、aac(6')-Ⅰb、tetA、tetK、cat2。且每株分离菌均检测出3种以上耐药基因。结果表明,分离的5株菌株均为绿脓杆菌,主要引起水貂出血性肺炎。分离菌株耐药性较严重,主要表现为多重耐药现象。菌株携带的耐药基因呈多样性,可引起菌株对相应药物产生耐药现象。     

Isolation and Identification of Local Aleutian Mink Virus Virulent Strain ADV-LN

Mink suspected infection aleutian mink disease virus (ADV) from mink breeding areas in Liaoning province were tested with CIEP method.The mink with antibody to ADV were selected and culled.Liver,spleen,kidney and mesenteric lymph node samples were taken for pathological examination and the viruses were observed under electron microscope.The grinded tissue fluid filter was added penicillin and treptomycin and inoculated into CRFK cells and passaged by 6 times for virus isolation.And cells cultures were identified as ADV by PCR.Then they were inoculated into healthy mink.Three days later,the mink showed clinical signs,which including the loss of appetite,anemia,hair dull,antifeedant and binge drinking.Some minks showed neurological symptoms,manifested symptoms of convulsions,cramps,staggering gait,ataxia,or hind limb paralysis and died.The virus strains isolated and identified were named as the ADV-LN.     

水貂志贺氏菌分离鉴定及药物敏感性分析

试验旨在对吉林省某水貂养殖场送检的3只具有典型腹泻症状的病死水貂的小肠和肠内容物样品进行细菌分离鉴定及耐药情况分析,为临床治疗提供参考。通过细菌分离纯化和PCR方法对分离菌株进行鉴定,对BALB/c小鼠进行菌液注射来检测菌株的致病性。采用K-B药敏法检测菌株对常用药物的敏感性,并通过PCR方法检测其耐药基因和Ⅰ类整合子的携带情况。结果显示,分离得到3株志贺氏菌,致病性检测试验显示分离菌可引起小鼠腹泻。药物敏感性试验结果显示,3株志贺氏菌对氟喹诺酮类药物、头孢类药物、氟苯尼考和多黏菌素较敏感,对氨基糖苷类、四环素、氯霉素、青霉素、氨苄西林耐药。耐药基因检测结果显示,3株志贺氏菌共检测出7种耐药基因bla_TEM-1、aadA1、aac（3'）-Ⅱc、aac（6'）-Ⅰb、aph（3'）-Ⅶ、tet（M）、cat2及携带aadA1基因盒的Ⅰ类整合子。结果表明,分离的3株志贺氏菌均具有致病性,可引起小鼠腹泻,主要表现为多重耐药现象,携带的耐药基因呈多样性,为临床治疗带来巨大影响。     

Persistent viral shedding during asymptomatic Aleutian mink disease parvoviral infection in a ferret.

A 2-year-old domestic ferret that appeared clinically healthy was repeatedly seropositive for Aleutian mink disease parvovirus (ADV) over a 2-year observation period. Antibody titers, determined by counter-immunoelectrophoresis, ranged from 1024 to 4096. Viral DNA also was identified in serum, urine, feces, and blood cell fractions by polymerase chain reaction analysis. Ultimately, DNA in situ hybridization revealed ADV DNA in histologic sections of various tissues and organs. These data indicate that this asymptomatic ferret was persistently infected with ADV.