鸡传染性贫血病疫苗免疫母鸡后子代雏鸡外周血液T细胞数量及免疫球蛋白含量变化

应用免疫SPA菌体不法和间接ELISE法对鸡传染性贫血病（CIA）疫苗免疫母鸡后，其子代雏鸡外周血液T细胞数量和血清IgG,IgM,IgA含量进行了动态研究。结果发现：CIA疫苗免疫母鸡后，其子代雏鸡外周血液T细胞数量和IgG,IgM,IgA含量均不同和蔼地高于未免疫的相对照雏鸡，表明CIA疫苗免疫母后，其子代雏鸡全身的体液上述各项指标均明显低于疫苗免疫的子代雏鸡，这与CIAV强毒攻击子代雏鸡后，未免疫的子代雏鸡外周血液上述各项指标均明显低于疫苗免疫的子代雏鸡，这与CIAV强毒感染雏鸡后，其免疫器官组织，尤其是胸腺严重损害，淋巴细胞变性坏死子等密切相关，是导致感染鸡免疫功能低下的基础。     

鸡传染性贫血疫苗免疫母鸡后子代雏鸡的免疫学变化

应用免疫学新技术对鸡传染性贫血（CIA）疫苗免疫母鸡后，其子代雏鸡外周血液和免疫器官组织及局部体液的免疫学变化进行了动态研究。结果表明：CIA疫苗免疫母鸡后，其子代雏鸡外周血液T、B细胞数量和IgG、IgM、IgA含量及免疫器官组织的T细胞和IgG、IgM、IgA抗体生成细胞以及泪液、气管液、胆汁、肠液的IgA、IgM、IgG含量均不同程度地高于未免疫的相应对照雏鸡，表明CIA疫苗免疫母鸡后，其子代雏鸡全身的体液免疫和细胞免疫功能明显增强。而CIAV强毒攻击后，未免疫的子代雏鸡，其外周血液和免疫器官组织及局部体液的上述各项指标均明显低于疫苗免疫的子代雏鸡。     

鸡传染性贫血病毒的分子生物学及其核酸检测技术

鸡传染性贫血 (CIA)是鸡的重要的免疫抑制性疾病之一 ,以引起雏鸡再生障碍性贫血和全身淋巴组织萎缩为主要特征。鸡传染性贫血病毒 (CIAV)主要引起鸡骨髓成红细胞和胸腺皮质的成淋巴细胞溶细胞感染 ,继而导致贫血和免疫抑制。该病毒在世界范围内广泛存在 ,是养鸡业潜藏的巨大威胁。文章从 CIA的危害、基本控制方法、病原的基因组及其所编码的蛋白质的功能、疾病的诊断等方面的最新进展进行了综述 ,重点介绍了病原的分子生物学研究进展及其核酸检测技术 ,并对 CIAV的研究前景进行了展望     

鸡贫血病毒实验室检测方法研究进展

鸡贫血病毒不仅可引起鸡的传染性贫血,而且也是引起鸡免疫抑制病的主要病原。鸡传染性贫血是以雏鸡再生障碍性贫血和免疫抑制为主要特征的传染病,是导致许多疫苗免疫失败以及雏鸡死亡的主要原因之一。鸡贫血病毒在世界范围内广泛存在,是养鸡业潜藏的巨大威胁。本文主要阐述了该病毒的病毒分离鉴定、血清学方法和分子生物学方法等实验室检测方法,旨在为鸡传染性贫血病的诊断和防治提供参考。     

鸡贫血病-法氏囊病联合免疫母鸡后子代雏鸡外周血液T细胞数量及免疫球蛋白含量变化

应用免疫SRA菌体花环法和间接ELISE法对鸡传染性贫血病 (CIA) -传染性法氏囊病 (IBD)联合免疫母鸡后 ,其子代雏鸡外周血液T细胞数量和免疫球蛋白IgG、IgM、IgA含量变化进行了动态研究。结果发现 ,CM -IBD联合免疫母鸡后 ,其子代雏鸡外周血液T细胞数量和IgG、IgM、IgA含量均不同程度地高于未免疫的相应对照雏鸡 ,表明CIA-IBD联合免疫母鸡后 ,其子代雏鸡外周血液体液免疫和细胞免疫功能明显增强。而CIAV、IBDY强毒攻击后 ,未免疫的子代雏鸡 ,其外周血液的上述指标均明显低于疫苗免疫的子代雏鸡 ,表明未免疫的子代雏鸡外周血液的免疫机能降低 ,这与未免疫雏鸡缺乏特异性抗体 ,强毒攻击后 ,雏鸡免疫器官组织广泛损害 ,淋巴细胞变性坏死等有关     

CIA-IBD联合免疫种鸡后子代雏鸡外周血液免疫学变化

应用免疫SPA菌体花环法、MTT法和间接ELISA法对鸡传染性贫血病(CIA)-传染性法氏囊病(IBD)联合免疫种鸡后,其子代雏鸡外周血液T细胞数量和增殖功能以及免疫球蛋白IgG、IgM、IgA含量变化进行了动态研究。结果发现,CIA-IBD联合免疫种鸡后,其子代雏鸡外周血液T细胞数量和IgG、IgM、IgA含量均不同程度地高于未免疫的相应对照雏鸡,表明CIA-IBD联合免疫种鸡后,其子代雏鸡外周血液体液免疫和细胞免疫功能明显增强。而CIAV、IBDV强毒攻击后,未免疫的子代雏鸡,其外周血液的上述指标均明显低于疫苗免疫的子代雏鸡,表明未免疫的子代雏鸡外周血液的免疫机能降低,这与未免疫雏鸡缺乏特异性抗体,强毒攻击后,雏鸡免疫器官组织广泛损害,淋巴细胞变性坏死等有关。     

云南省地方品种鸡群4种垂直传播病毒血清流行病学调查

为了解云南省地方品种鸡群中禽白血病病毒(ALV)、禽网状内皮组织增生症病毒(REV)、鸡传染性贫血病毒(CAV)和禽脑脊髓炎病毒(AEV)的感染情况,采用ELISA方法对云南省滇东、滇西、滇中、滇南和滇北5个区域8个地方品种鸡群的2295份血清样品进行了上述4种垂直传播病毒的抗体检测。结果显示:云南省8个地方品种鸡群中,ALV、REV、CAV和AEV平均抗体阳性率分别为27.02%、72.77%、98.47%和76.25%,二重、三重、四重感染率分别为32.90%、36.38%、15.90%;不同区域、不同品种鸡群中均存在上述4种病原的感染,抗体阳性率有一定差异,但无规律性。结果表明,云南省地方品种鸡群中4种垂直传播病毒感染率较高,流行面较广,且呈现严重的混合感染状态。结果提示:应重视这4种可垂直传播病原的流行控制,谨防因病毒混合感染导致重大疫病免疫失败;继续加强抗体检测,及时淘汰阳性鸡群,净化种群,同时定期进行疫苗污染监测,提升饲养管理水平,保障云南省地方鸡种质资源的安全健康发展。     

Detection of pigeon circovirus by polymerase chain reaction

Pigeon circovirus was identified by polymerase chain reaction (PCR) in young pigeons belonging to 12 different lofts. Viral DNA was extracted from formalin-fed, paraffin-imbedded tissues containing primarily bursa and occasionally liver and spleen with a commercial kit. PCR primers were selected from a published sequence for columbid circovirus and evaluated in a PCR assay. The histopathologic examination of various tissues revealed basophilic globular intracytoplasmic inclusions in the mononuclear cells of the bursa of Fabricius and occasionally in the spleen characteristic for a circovirus. Transmission electron microscopy of a few bursas of Fabricius revealed virus particles measuring 18-21 nm. All the samples were negative by PCR for psittacine beak and feather disease (PBFD) virus and chicken infectious anemia virus. The primers for both pigeon circovirus and PBFD virus did not react in PCR with the chicken anemia virus DNA. Most of the circovirus-infected pigeons had concurrent infections of Escherichia coli, Salmonella, Pasteurella, Aspergillus, candidiasis, nematodiasis, or capillariasis.     

鸡传染性贫血免疫对肝微粒体抗氧化酶活性的影响

目的是观察鸡传染性贫血多次免疫对肝微粒体中抗氧化酶活性的影响.对20只SPF鸡随机分为2组,每组10只,免疫组鸡用鸡传染性贫血弱毒苗免疫4次,每次间隔2周,对照组鸡注射同剂量的生理盐水.最后一次免疫后10d取肝脏制备微粒体,利用测试盒测定肝微粒体中的GSH-Px活性、SOD活性、CAT活性和MDA含量.结果与对照组相比,免疫组肝微粒体中GSH Px活性、SOD活性和CAT活性都显著提高(P＜0.05),MDA含量显著减低(P＜0.05).结论为鸡传染性贫血多次免疫可提高鸡体抗氧化能力.     

Isolation and identification of chicken infectious anemia virus in Brazil.

Seven chicken infectious anemia virus (CIAV) isolates were obtained from seven broiler flocks with poor performance in two states of Brazil. All isolates induced thymus atrophy, bone-marrow aplasia, and low hematocrit values when inoculated into 1-day-old susceptible chicks. The CIAV isolates were resistant to treatment with chloroform and were able to pass through 50-nm-pore-size filters. CIAV-specific antigens could be demonstrated in tissues of experimentally infected chicks using a monoclonal antibody specific for CIAV. These characteristics of the virus and the virus-induced lesions demonstrate that CIAV is present in Brazil and that the virus is associated with production problems.     

Partial characterization of an adenovirus-like virus isolated from broiler chickens with transmissible viral proventriculitis

Transmissible viral proventriculitis (TVP) was experimentally reproduced in specific-pathogen-free chickens using a homogenate of proventricular tissue obtained from TVP-affected commercial broiler chickens. Thin-section electron microscopy revealed intranuclear, approximately 70-nanometer (nm), adenovirus-like viruses (AdLV) within proventricular lesions. The AdLV, designated AdLV (R11/3), could not be propagated using various avian and mammalian cell cultures or by inoculation of embryonated chicken eggs by yolk, allantoic, or chorioallantoic membrane routes. However, AdLV (R11/3) was successfully propagated by amniotic inoculation of embryonated chicken eggs, with detection of the virus in proventriculi and intestinal contents of hatched 2-day-old chicks (8 days postinoculation). Virus propagation was evident in in ovo-inoculated chicks by (1) gross and microscopic lesions in proventriculi consistent with TVP, (2) immunohistochemical localization of AdLV (R11/3) antigens in proventricular epithelium, (3) thin-section electron microscopic detection of intranuclear, approximately 70-nm AdLVs within proventricular epithelium, and (4) negative-stain electron microscopic detection of extracellular, approximately 70-nm AdLVs in intestinal contents. Indirect immunofluorescence and polymerase chain reaction procedures that specifically recognize groups I, II, and III avian adenoviruses failed to recognize AdLV (R11/3). The findings suggest an etiologic role for AdLV (R11/3) in TVP and indicate that this virus is distinct from known avian adenoviruses.     

Economically Important Non-oncogenic Immunosuppressive Viral Diseases of Chicken—Current Status

Immunosuppressive viral diseases threaten the poultry industry by causing heavy mortality and economic loss of production, often as a result of the chickens' increased susceptibility to secondary infections and sub-optimal response to vaccinations. This paper aimed to present an up-to-date review of three specific economically important non-oncogenic immunosuppressive viral diseases of chickens, viz. chicken infectious anaemia (CIA), infectious bursal disease (IBD) and hydropericardium syndrome (HPS), with emphasis on their immunosuppressive effects. CIA and IBD causes immunosuppression in chickens and the socio-economic significance of these diseases is considerable worldwide. CIA occurs following transovarian transmission of chicken anaemia virus and has potential for inducing immunosuppression alone or in combination with other infectious agents, and is characterized by generalized lymphoid atrophy, increased mortality and severe anemia. The virus replicates in erythroid and lymphoid progenitor cells, causing inapparent, sub-clinical infections that lead to depletion of these cells with consequent immunosuppressive effects. The IBD virus replicates extensively in IgM(+) cells of the bursa and chickens may die during the acute phase of the disease, although IBD virus-induced mortality is highly variable and depends, among other factors, upon the virulence of the virus strain. The sub-clinical form is more common than clinical IBD because of regular vaccination on breeding farms. Infection at an early age significantly compromises the humoral and local immune responses of chickens because of the direct effect of B cells or their precursors. HPS is a recently emerged immunosuppressive disease of 3-6-weeked broilers, characterized by sudden onset, high mortality, typical hydropericardium and enlarged mottled and friable livers, with intranuclear inclusion bodies in the hepatocytes. The agent, fowl adenovirus-4, causes immunosuppression by damaging lymphoid tissues; the presence of IBD and CIA viruses may predispose for HPS or HPS may predispose for other viral infections. Synergism with CIA or other virus infections or prior immunosuppression is necessary to produce IBH-HPS in chickens and the susceptibility of chickens infected with fowl adenovirus varies throughout the course of CIA infection. The mechanism of immunosuppression has been studied in detail for certain chicken viruses at molecular levels, which will provides new opportunities to control these diseases by vaccination.     

Isolation and preliminary characterization of chicken anemia virus from chickens in Nigeria

Chicken anemia virus (CAV) was isolated for the first time from the Nigerian chicken population. The virus was recovered from necropsied birds from broiler and pullet flocks that suffered disease outbreaks tentatively diagnosed as infectious bursal disease. A sensitive polymerase chain reaction (PCR) assay detected CAV DNA in tissues of necropsied birds. Restriction endonuclease analysis performed with the 733-bp PCR product and the Cfo I enzyme indicated at least two different CAVs were circulating among the Nigerian chicken population. Four isolates were obtained from pooled liver and thymus tissues using the MDCC-MSB1 cell line. These isolates were found to be antigenically closely related to the Cuxhaven-1 (Cux-1) reference strain of CAV when reacted with four monoclonal antibodies prepared against the Cux-1 virus. One of the isolates (isolate A) induced thymus atrophy, bone marrow aplasia, and low hematocrit values when inoculated into 1-day-old specific-pathogen-free chickens. These findings not only demonstrate that CAV is present in Nigeria, but they also likely represent the first cell culture isolation of the virus in Africa.     

应用聚合酶链反应检测鸡传染性贫血病毒

根据鸡传染性贫血病毒（CAV）Cux－1株的基因序列,设计了一对引物,用这对引物对三株CAV的核酸模板进行PCR扩增,结果均能特异性地扩增出420bp的片段,但对其它5种禽病病原体核酸模板的扩增,结果均为阴性,该PCR能检出10fg鸡传染性贫血病毒DNA模板。     

The isolation and characterization of chicken anemia agent (CAA) from broilers in the United States

A chicken anemia agent (CAA) isolated from commercial broilers in the United States was characterized in vivo and in vitro. When inoculated into susceptible 1-day-old chickens, the agent induced a severe bone marrow aplasia, thymic atrophy, multiple subcutaneous and intramuscular hemorrhages, and anemia, as evidenced by reduced hematocrits. Chickens derived from different breeder flocks and inoculated in ovo or at 1 day of age varied in their susceptibility to the CAA, with some flocks being highly susceptible, while others were almost totally resistant. This was true for both specific-pathogen-free and commercial chickens. The isolate was able to pass through a 50-nm-pore-size filter and was resistant to inactivation at 56 C for 30 minutes. It failed to agglutinate avian and mammalian erythrocytes and could not be propagated in conventional cell cultures. The physical and biological characteristics of the agent and the disease it induces indicate that it is similar to the CAA found in Japan and Europe.     

Serological evidence of chicken infectious anemia virus in the United States at least since 1959

A retrospective, serological survey was performed to determine an approximate time frame for when chickens were first exposed to chicken anemia virus (CAV) in the southeastern United States. A serum collection covering most of the period between 1959 and 2005 was available for the present study. These sera were obtained from adult chicken flocks that were maintained in experimental chicken farms at Auburn University's Department of Poultry Science. Sera were tested for the presence of CAV-specific antibodies using a commercially available competitive enzyme-linked immunosorbent assay (ELISA) kit. Values <0.6 were considered positive. Fresh sera obtained from hens in 2005 showed 45.5% negative and 54.5% positive for CAV antibodies. The assessment of serum samples covering the time period of 1959 through 1979 resulted in most sera being positive for CAV antibodies. The percentage of positive samples between years varied from 43% to 100%. These serological results support assumptions based on circumstantial evidence that CAV must have been present in the United States long before its first isolation in 1989.     

Polymerase chain reaction amplification for direct detection of chicken anemia virus DNA in tissues and sera.

Direct detection of chicken anemia virus (CAV) DNA in tissues and sera was investigated by a polymerase chain reaction (PCR) assay. Using a pair of primers constructed to amplify the coding sequence of the CAV DNA genome, the PCR assay was shown to be extremely sensitive, being able to detect 1 fg of CAV replicative form DNA. The oligonucleotide primers used for the PCR yielded 583 base-pair (bp) amplified product, which was sized by ethidium bromide-agarose gel electrophoresis. Tissue samples from seven cases of suspected chicken infectious anemia were obtained for CAV isolation. DNA extracted from the homogenized suspension of pooled tissues of each case was analyzed by the PCR assay. Results showed that five of the seven cases were positive for CAV DNA by PCR, whereas CAV was isolated from four cases only. The PCR assay also detected CAV DNA in two of 37 serum samples from disease-free chickens. The specificity of PCR was confirmed by chemiluminescence dot-blot analysis of the amplified products.     

我国部分地区蛋鸡群ALV-J及与REV、MDV、CAV混合感染检测

为了解J亚群禽白血病病毒(ALV-J)及其与禽网状内皮细胞增生症病毒(REV)、马立克氏病病毒(MDV)和鸡传染性贫血病病毒(CAV)的混合感染现象,本研究从宁夏、湖北、广东、山东、辽宁、吉林、黑龙江7个省的39个蛋鸡群收集临床表现和剖检病理变化疑似禽白血病的病料样品184份,采用PCR、病毒分离和IFA检测样品中ALV-J、REV、MDV和CAV。结果表明,7个省蛋鸡场均存在ALV-J感染,病料样品阳性率为60.9%,检测鸡群阳性率为82.1%,与REV、MDV、CAV的混合感染率分别为13.6%、24.5%、22.8%,其中存在较为严重的双重感染(29.0%)和3重感染(18.8%),甚至4重感染(1.7%)。研究结果表明,我国蛋鸡群中普遍存在ALV-J感染,而且与REV、MDV、CAV混合感染严重;提示ALV-J已经可以引起蛋鸡群发病,在临床诊断和致病性研究中,应考虑到多重感染的影响。     

鸡贫血病—法氏囊病疫苗免疫母鸡后子代雏鸡的免疫学变化

本项目应用现代免疫学新技术对鸡传染性贫血病（CIA）-传染性法氏囊病（IBD）疫苗联合免疫母鸡后,其子代雏鸡外周血液T、B细胞数量和IgG、IgM、IgA含量法及法氏囊、胸腺、脾脏、盲肠扁桃体、哈德尔腺的T细胞和IgG、IgM、IgA抗体生成细胞数量以及泪液、气管液、胆汁、肠液的IgA、IgM、IgG含量的变化进行了动态研究。结果发现,CIA-IBD疫苗联合免疫母鸡后,其子代雏鸡外周血液、免疫器官组织和局部体液的上述各项指标均不同程度地高于未免疫的相应对照雏鸡。表明CIA-IBD疫苗免疫母鸡后,其子代雏鸡的体液免疫和细胞免疫功能明显增强,而CIAV-IBDV强毒攻击后,未免疫的子代雏鸡,其外周血液,免疫器官组织和局部体液的各项免疫学指标均明显低于疫苗免疫攻毒的子代雏鸡,这与未免疫雏鸡缺乏特异性抗体,强毒攻击后,雏鸡免疫器官组织广泛损害,淋巴细胞变性坏死等有关。     

Virus Pneumonia of Pigs; Attempts at Propagation of the Causative Agent in Cell Cultures and Chicken Embryos

Two strains of the agent of virus pneumonia, were tested for the ability to propagate in 12 types of cell cultures and in chicken embryos. The 5 primary cell cultures used were: swine kidney, lung, bone marrow, testicle, and chicken embryo kidney; and the 7 serial passage cell cultures were: swine kidney, kidney-tumor, testicle, bone-marrow, bovine kidney, and human cervical carcinoma (HeLa). The agent of virus pneumonia was propagated in primary swine kidney and in HeLa cell cultures as shown by the production of typical gross and microscopic lesions in pigs inoculated with cell future fluids. Third passage cell culture fluids, produced typical gross lesions in pigs, but fourth passage cell culture fluids produced only microscopic lesions, and no lesions were produced by sixth and eleventh passage fluids. Control pigs receiving fluids from uninoculated cell cultures remained free of gross or microscopic lesions, as did uninoculated controls. Cytopathic effects were not detected in any of the inoculated cell cultures and no cellular changes were detected by staining with Giemsa stain or acridine orange.

Neither lesions nor deaths occurred in chicken embryos inoculated with both strains of virus pneumonia virus. Pneumonia was not produced in pigs inoculated with suspensions from second chicken embryo passage of the 2 strains inoculated by the chorioallantioic sac, the amniotic sac, and the yolk sac routes.

Identical gross and microscopic lesions were produced in pigs inoculated with either pneumonic lung suspensions or with virulent cell culture fluids. Gross lesions consisted of areas of light to reddish-purple consolidation usually limited to the anterior, cardiac, and intermediate lobes of the lungs. Pleuritis and pericarditis were never present in experimentally produced virus pneumonia. The microscopic lesions were characterized by: 1. perivascular and peribronchiolar lymphoid infiltration and hyperplasia, 2. alveolar interstitial thickening and infiltration, and 3. alveolar exudates consisting of alveolar cells, lymphocytes, plasma cells, and neutrophiles.