祁连县鹿场鹿布鲁氏菌病调查

布鲁氏菌病 (以下简称布病 )是由布鲁氏菌引起的人畜共患传染病。为了解青海省祁连县鹿场鹿的布病感染及流行情况 ,笔者借结核病检疫的机会进行了临床调查 ,并于 2 0 0 1年 11月对祁连县鹿场的 2 0 6份鹿血样进行了布病血清学检验。现将调查情况报告如下 :1　基本情况祁连县鹿场始建于 1964年 ,地处祁连县野牛沟乡境内 ,距县城 40ｋｍ ,海拔高度 3 0 0 0～ 3 3 0 0ｍ ,总面积 2 6ｋｍ2 ,分割围栏12个 ,以灌丛草场为主。现有职工 3 2人 ,饲养各类鹿 3 80头 ,其中梅花鹿 60头、白唇鹿 13 8头、马鹿 182头 ,并养有少量牦牛。2　材料与方法2 1…     

白唇鹿结核病的病原分离

从 1998年开始 ,青海省祁连鹿场白唇鹿群发生以慢性消瘦、肺部有结节为特征的疫病 ,以后逐年增加 ,至 2 0 0 1年已死亡鹿 2 0 0余只。 2 0 0 1年 3月 ,捕杀祁连鹿场的 3只病鹿 ,无菌采取有病变的肺组织和淋巴结进行病原分离。1　材料与方法1.1　材料　　病料采自青海省祁连鹿场病死鹿 ;实验动物 (豚鼠 )购自青海省兽医生物药品厂动物房 ;选用的改良小川氏、杜布氏、杜赛氏培养基按参照文献 [1]自制。1.2　方法1.2 .1　抗酸染色　参照文献 [2 ]进行。1.2 .2　病原分离　材料处理参照文献 [2 ]进行 ,病料经青霉素处理后 ,接种于上述培养基 ,37…     

鹿副结核病抗体间接ELISA检测方法的建立

本研究以副结核杆菌亲和层析抗原为检测抗原,检测以草分枝杆菌抗原吸收的待检鹿血清,建立检测鹿副结核病血清抗体的间接酶联免疫吸附试验,确定其抗原最佳包被浓度为40μg/mL,血清样品稀释度为1:80,兔抗鹿IgG辣根过氧化物酶标记抗体稀释度为1:8000。经特异性试验和重复性试验证明该方法特异性高、重复性好。对不同地区4个鹿场的760头份鹿血清进行副结核病抗体检测,其中阳性61头份,阳性率为8%,获得副结核病在我国鹿群中的血清流行病学资料,从而为防制鹿副结核病提供一定的依据。     

麻鹿和梅花鹿肠毒血症的诊断与防治

鹿肠毒血症是由D型魏氏梭菌引起的一种急性肠毒血症 ,病的发生是由于魏氏梭菌在肠道内大量繁殖 ,产生毒素所引起。魏氏梭菌根据毒素 -抗毒素中和试验分为A、B、C、D、E五型。每型产生一种主要毒素 ,一种或数种次要毒素。肠毒血症是由D型魏氏梭菌引起的一种疾病。1　流行情况宁夏中卫某鹿场原有 32头麻鹿和梅花鹿。 2 0 0 1年 6月 ,从甘肃某鹿场调入 4 5头两种鹿 ,8月中旬 ,又从该鹿场调入 79头两种鹿 ,使该鹿场鹿群数达 15 6头。从 6月份鹿场调入鹿群后不久 ,在鹿群中有不同年龄的鹿发生症状相似的病 ,最急性无任何症状突然发病 ,多在 4…     

后海穴注射庆大霉素治疗仔鹿腹泻

仔鹿腹泻是初生仔鹿的一种常见病和多发病 ,疾病发生原因主要是母鹿乳房或饲料饮水污染 ,人工哺乳不当引起的一种炎性疾病 ,此病治愈率为80 %左右 ,死亡率高达 1 8%～ 2 0 %。笔者自 1 994年以来采用庆大霉素后海穴注射效果明显 ,共收治 42例 ,治愈 3 9例。1　病例　收治 42例病畜均为新疆塔河马鹿。分别在新疆兵团农二师 3 2团一鹿场 2 2例 ,二鹿场 8例 ,三鹿场 1 2例 ,3 6例发病后 2天内就诊 ,余 6例 2天后诊治 ,因脱水死亡 3例。2　临床症状　疾病多发于 2 0～ 6 0日龄的初生仔鹿。主要是腹泻、拉稀 ,可见粪便稀薄至水样 ,呈污褐色 ,有时…     

鹿蠕形螨病诊治

延吉市某鹿场饲养627只鹿。因2008年5月初鹿场发生鹿的皮肤病，该鹿场邀请就诊。通过临床症状和流行病学调查，结合实验室检查，确诊为鹿蠕形螨病。     

仔鹿下痢原因分析及综合防治实践

仔鹿是鹿场发展的希望,是优良种群的基础,是培育高产性状的关键。 仔鹿下痢(腹泻)是养鹿生产中普遍存在的棘手问题,任何一个鹿场都回避不了。 首先,仔鹿下痢是导致仔鹿死亡的主要原因,造成鹿场直接经济损失,一些鹿场仔鹿成活率不到50％,甚至更低。     

鹿快疫的诊断与防制

1997年 7月中旬 ,我市某养鹿场 ,发生了一种急性传染病。该病发病急 ,病程短 ,突然死亡 ,真胃粘膜呈现出血性坏死性炎症。根据流行病学调查、临床特征、病理剖检、实验室诊断 ,确诊为鹿快疫 ,现报告如下。1　发病情况及临术症状　该鹿场于 1 998年初引进马鹿 6 0只 ,其中母鹿 45只 ,公鹿 1 5只 ,自繁仔鹿 2 0只 ,截止 1 999年1 0月 ,鹿场存栏马鹿 80只。 1 999年 5月、8月鹿场突然二次死亡 2只鹿 ,1 0月又突然死亡 1只 ,其中 2只 2岁 ,1只 1 .5岁。有 2只夜间突然发病死亡 ,1只白天吃草时突然发病 ,表现运动失调 ,口吐白沫 ,全身震颤 ,四肢…     

鹿酒糟中毒的诊治

<正>酒糟中毒是由于家畜长期采食酒糟而引起的中毒,会造成养鹿业巨大经济损失。2018年2月吉林省抚松县松花江鹿场仔鹿突发疾病,通过病理剖检、实验室检测方法确诊为鹿酒糟中毒,根据鹿酒糟中毒症状发现与其他反刍动物中毒的症状相似。报告如下。1 发病情况吉林省抚松松花江鹿场饲养成年母鹿500头,公鹿100头,在2018年2月该鹿场仔鹿出现起立难,食欲不振现象,发病时当地出现阴雨天气,电话询问初步判断为白肌病,遂使用亚硒酸钠、磺胺类药物治疗,效     

饲喂微量元素饲料添加剂增产鹿茸的效果

为摸索鹿茸增产途径,增加养鹿生产经济效益,引龙河农场畜牧公司鹿场于1982年3—6月,进行了对鹿饲喂微量元素饲料添加剂增产鹿茸的试验,收到较好的效果。一、材料与方法选本公司鹿场1—9锯龄生产公鹿     

马立克氏病病毒“814”疫苗株基因组重复区序列测定及分析

为了解马立克氏病病毒(MDV)强、弱毒株主要致肿瘤相关基因变异情况,本研究根据MDV GA株基因组序列,设计合成扩增基因组重复区的引物,得到MDV814疫苗株病毒基因组中约26kb的序列片段。与GenBank登录的强、弱毒株进行比较分析表明,扩增的MDV1型814疫苗株的长重复区为12774bp,预测的开放阅读框(ORF)有48个;短重复区为11426bp,预测的ORF有38个。发现了4个MDV814株特有的ORF。814株在编码Meq、RLORF6和23ku的重叠基因内具有类似于疫苗株CVI988的177bp的插入;在RLORF12基因编码区内存在69bp的缺失,该缺失位于病毒复制起始位点内。同时,发现7个814疫苗株特有的氨基酸突变,分布在6个ORF内。单核苷酸多态性(SNP)的鉴定发现,多个基因具有单核苷酸的突变,主要分布于Meq基因,其中氨基酸A115V(丙氨酸-缬氨酸),N142D(天冬酰胺-天门冬氨酸)的变异是814疫苗株所特有的。MDV814疫苗株重复区的基因序列的比较分析将有助于MDV致肿瘤机制的研究。     

Identification and characterization of Marek's disease virus serotype 1 (MDV1) ICP22 gene product: MDV1 ICP22 transactivates the MDV1 ICP27 promoter synergistically with MDV1 ICP4

A previous report [Virus Genes 6 (1992) 365-378] has shown that the US1 gene of Marek's disease virus serotype 1 (MDV1) encodes a homologue of herpes simplex virus type 1 infected cell protein No. 22 (ICP22). In the present study, we expressed and identified a product of the MDV1 US1 gene in chicken embryo fibroblasts (CEFs) with the aid of a recombinant baculovirus expressing a Flag epitope-tagged MDV1 US1 gene, under control of the SRalpha promoter (composed of the enhancer region of the simian virus 40 early promoter and the R region of the human T-cell leukaemia virus type 1 long terminal repeat). In CEF infected with the recombinant baculovirus, MDV1 ICP22 was specifically and efficiently expressed in the presence of n-butyric acid. The apparent M(r) of the expressed protein was 30,000. Reporter gene assays revealed that MDV1 ICP22 by itself transactivated an MDV1 ICP27 promoter/reporter construct weakly but specifically, and furthermore, worked synergistically with MDV1 ICP4 to efficiently up-regulate the MDV1 ICP27 promoter. MDV1 ICP22 may be a regulatory protein that stimulates viral promoters in co-operation with other viral regulatory proteins such as MDV1 ICP4.     

Viral pathogenesis in chicken embryos and tumor induction in chickens after in ovo exposure to serotype 1 Marek's disease virus

We examined the susceptibility of late-stage chicken embryos to infection with oncogenic serotype 1 Marek's disease virus (MDV 1). Intravenous inoculation of MDV 1 at embryonic day (ED) 16 resulted in significant replication of the virus in embryonic tissues. Within 5 days of virus exposure, pp38 viral antigen (pp38) was detected in embryonic bursae and MDV 1 was isolated by plaque assay from the spleens, thymuses, and bursae of embryos. The pathogenesis of MDV 1 after intravenous inoculation at ED 16 was similar to that in chicks exposed to MDV 1 after hatching. In contrast to the response of the embryo to intravenous inoculation, embryos exposed to MDV 1 by the amniotic route did not develop detectable pp38, nor could the virus be isolated from the embryonic tissues by plaque assay. These results show that the route of inoculation of MDV 1 in the embryos is critical for allowing the virus to come in contact with target cells.     

Delayed replication of Marek's disease virus following in ovo inoculation during late stages of embryonal development

Several oncogenic and non-oncogenic isolates of Marek's disease virus (MDV) were inoculated into embryonated eggs on embryonation day (ED) 16 to 18, and embryos or chicks hatching from inoculated eggs were examined for infectious virus and viral internal antigen (VIA) in lymphoid organs. There was no evidence of extensive replication of MDV in any of the embryonic tissues examined. Levels of VIA peaked 4-5 days after chicks hatched. This indicated that MDV remained inactive during embryonation and did not initiate pathogenic events until chicks hatched. Because HVT replicated rapidly in the embryo but MDV did not, in ovo inoculation of HVT simultaneously with oncogenic MDV or several days after MDV resulted in significant protection (P less than 0.025) of hatched chicks against Marek's disease (MD). Little protection was obtained if HVT was given simultaneously with MDV or after MDV to chicks already hatched. The relative susceptibility of the embryo to extensive replication of the vaccine virus but not the challenge virus apparently accounted for protection against MD in chicks hatching from dually infected eggs.     

Development of a nested polymerase chain reaction method to detect oncogenic Marek's disease virus from feather tips.

For the easy survey of Marek's disease virus (MDV), feather tip-derived DNA from MDV-infected chickens can be used because feather tips are easy to collect and feather follicle epithelium is known to be the only site of productive replication of cell-free MDV. To develop a diagnostic method to differentiate highly virulent strains of MDV from the attenuated MDV vaccine strain, CVI988, which is widely used, nested polymerase chain reaction (PCR) was performed to detect a segment of the meq gene in feather tip samples of chickens experimentally infected with MDV. In chickens infected with Md5, a strain of oncogenic MDV, the meq gene was consistently detected, whereas the L-meq gene, in which a 180-base pair (180-bp) sequence is inserted into the meq gene, was detected in CVI988-infected chickens. Moreover, the meq gene was mainly detected even in chickens co-infected with both Md5 and CVI988. These results suggest that this method is appropriate for the surveillance of the highly virulent MDV infection in the field.     

Marek's disease virus infection in the brain: virus replication, cellular infiltration, and major histocompatibility complex antigen expression

Marek's disease virus (MDV) infection in the brain was studied chronologically after inoculating 3-week-old chickens of two genetic lines with two strains of serotype I MDV representing two pathotypes (v and vv+). Viral replication in the brain was strongly associated with the development of lesions. Three viral antigens (pp38, gB, and meq) were detected in the brain of infected chickens. Marked differences between v and vv+ pathotypes of MDV were identified for level of virus replication, time course of brain lesions, and expression of major histocompatibility complex (MHC) antigens. Two pathologic phenomena (inflammatory and proliferative) were detected in the brain of chickens inoculated with vv+MDV, but only inflammatory lesions were observed in those inoculated with vMDV. Inflammatory lesions, mainly composed of macrophages, CD4+ T cells, and CD8+ T cells, started at 6-10 days postinoculation (dpi) and were transient. Proliferative lesions, characterized by severe infiltrates of CD4+CD8- T cells (blasts), started at 19-26 dpi and persisted. Expression of MHC antigens in endothelial cells and infiltrating cells within the brain was influenced by MDV infection. Upregulation of MHC class II antigen occurred in all treatment groups, although it was more severe in those inoculated with vv+MDV. MHC class I antigen was downregulated only in those groups inoculated with vv+MDV. These results enhance our understanding of the nature and pattern of MDV infection in the brain and help to explain the neurovirulence associated with highly virulent MDV.     

Identification of the cross-reactive antigens between Marek's disease virus and pseudorabies virus by monoclonal antibodies

Among the 33 monoclonal antibodies (MAbs) against pseudorabies virus (PRV) examined, three MAbs (24-17, 74-26, and 8) were found to react with cells infected with Marek's disease virus (MDV)-related viruses by immunofluorescence test. Two of the MAbs (24-17 and 74-26) reacted with the nuclei of cells infected with MDV serotype 1 (MDV1), MDV serotype 2 (MDV2), and herpesvirus of turkeys (HVT), whereas MAb 8 reacted with the cytoplasm of MDV2- and HVT-infected cells. However, none of the MAbs against MDV1, MDV2, and HVT that were examined reacted with PRV-infected cells. None of these three MAbs against PRV reactive with MDV-related viruses cross-reacted with the cells infected with other herpesviruses, such as herpes simplex virus type 1, herpes simplex virus type 2, varicella zoster virus, Epstein-Barr virus, or human herpesvirus 6. Southern-blot hybridization under stringent or less-stringent conditions showed that no significant DNA homology was detected between PRV DNA and MDV DNA.     

人参皂苷及其衍生物体内抗马立克氏病毒的间接免疫荧光观察与PCR检测

为了探讨人参皂苷及其衍生物体内抗马立克氏病毒的作用机理.用马立克氏病毒人工感染雏鸡模型,采取人参皂苷及其衍生物口服,通过间接免疫荧光试验动态观察人参皂苷及其衍生物能否减少MDV抗原在组织中的分布;通过PCR检测看人参皂苷及其衍生物能否减少MDV的出现.结果显示:人参皂苷组、衍生物组被检组织的阳性细胞数量明显比盐酸吗啉胍阳性对照组被检组织的阳性细胞数量少;MDV病毒核酸的PCR检测显示,药物没有阻止病毒对组织的感染.结果表明,人参皂苷及其衍生物抗马立克氏病毒效果要优于盐酸吗啉胍.     

应用双重实时荧光定量PCR方法检测鸡马立克氏病血清1型病毒

本研究建立了鸡马立克氏病血清1型病毒（MDV1）绝对定量检测方法。研究中选择MDV1特有的Meq基因的一段保守序列作为检测对象，将其克隆到质粒载体中，作为阳性标准品；同时将管家基因．鸡卵铁转蛋白（Ovo）特异性基因片段克隆到质粒载体上作为内参照的标准品。经荧光定量PCR（FQ-PCR）法扩增获得MDV1的FQ-PCR两条标准曲线，建立了MDV1双重FQ-PCR检测方法。应用该方法绝对定量检测了实验攻毒鸡及吉林省某地发病鸡只的羽髓、淋巴细胞等组织样本中单位细胞病毒拷贝数，并与琼脂扩散（AGP）、常规PCR等检测方法进行比较。结果表明，不论实验攻毒鸡还是自然发病鸡，羽髓中病毒富含量均高于其它组织，每百万宿主细胞内病毒含量为10^7～10^8拷贝；FQ-PCR检测MD发病鸡只的阳性率高于AGP，达100％；该方法的灵敏度比常规PCR检测高10～100倍，在单位细胞内可灵敏地检测到2．78个拷贝的病毒。该方法可以在不同的样品中有效的绝对定量检测MDVl。