安徽省鸡免疫抑制性疾病的流行病学调查

采集了安徽省6个主要养鸡地区65群鸡的185只病鸡共986份组织样品，对可引起免疫抑制性疾病的5种最常见病毒进行了PCR检测。结果，传染性腔上囊病病毒（IBDV）、鸡传染性贫血病毒（CIAV）、马立克氏病病毒（MDV）、禽白血病病毒（ALV）和网状内皮增生症病毒（REV）的群阳性率和个体阳性率分别为73．08％和40．91％、46．15％和25．95％、41．54％和17．84％、18．46％和7．57％、13．85％和4．86％，其中被检鸡之二重或多重混合感染的总阳性率为24．33％。调查结果证实，免疫抑制性疾病在安徽省的商业鸡群中普遍存在，并与鸡群疾病多而复杂、损失大相关。     

鸡的免疫抑制性疾病及解决措施

鸡传染性贫血（CIAV）、网状内皮细胞增生（REV）、淋巴性白血病（ALV-JALV-A亚型）、法氏囊（IBDV）、马立克（MDV）等是近年来鸡群中较常见的免疫抑制性疾病,尤其以CIAV、REV、ALV—J感染较为严重,对养鸡生产的危害已引起国内外的高度关注。以我国著名禽病学家崔治中教授为代表的禽病学者对鸡免疫抑制性疾病展开了较为深入的研究。研究表明,免疫抑制病多为亚临床感染,导致鸡体免疫器官（如法氏囊、胸腺等）受损甚至萎缩,     

鸡传染性贫血病的研究进展

鸡传染性贫血病（CIA）是由鸡传染性贫血病毒（CIAV）引起的一种以雏鸡再生障碍性贫血和全身淋巴组织萎缩为主要特征的病毒性传染病。它和马立克氏病病毒（MDV）、传染性法氏囊病病毒（IBDV）、网状内皮增生症病毒（REV）、禽白血病病毒（ALV）和禽呼肠孤病毒（ARV）是危害养鸡业最常见的免疫抑制性病毒。     

我国商业鸡群多种免疫抑制性病毒共感染的研究

在2000～2006年间通过对来自我国4个主要养鸡省区的193个商业鸡群共808只病/死鸡分别进行5种免疫抑制性病毒CIAV、IBDV、MDV、REV和ALV感染情况的调查,结果显示:CIAV、IBDV、MDV、ALV和REV的群阳性率和个体阳性率分别为65.56%和30.94%、49.14%和23.98%、35.63%和12.5%、12.22%和4.29%、12.22%和4.16%,其中被检鸡二重及多重混合感染的总阳性率为31.73%,最常见的共感染形式是MDV CIAV(阳性率为10.02%)和MDV ALV(阳性率为8.50%);7个不同品种鸡被各种病毒的感染率也不尽相同。结果证实,免疫抑制性病毒的感染在4个省区的商业鸡群中普遍存在。     

医学我国鸡群中免疫抑制性病毒多重感染的诊断和对策

我国雏鸡和青年鸡群中,由不同的免疫抑制性病毒多重感染诱发的免疫抑制性疾病越来越常见,由它造成的经济损失日趋严重.有许多病毒感染都可能造成鸡群不同程度的免疫抑制状态,如传染性法氏囊病病毒(IBDV),马立克病病毒(MDV),网状内皮增生病病毒(REV),鸡传染性贫血病毒(CIAV),呼肠孤病毒(ReoV,传染性关节炎病毒), 鸡白血病病毒(ALV)等.在这类可引起鸡群免疫抑制的病毒感染中,网状内皮增生病病毒、鸡传染性贫血病毒和鸡传染性关节炎病毒(吸肠毒,ReoV)这三种病毒不仅能通过接触横向传染,还能通过鸡胚垂直传染.而且这类病毒对雏鸡、特别是一日龄雏鸡最易感.因此, 如果在用于制造弱毒疫苗的一批鸡胚中,有个别污染了这类病毒,就会使整批疫苗实际上都污染了病毒,导致给出壳雏鸡免疫接种时人为地造成全群感染.这样的事故,即使在国外发达国家也时而有报道[1].鸡群的免疫抑制性病毒多重感染不仅影响鸡群的生产性能,更易导致多种其它不同的细菌性和病毒性继发性感染,且易造成对特定疫苗免疫反应的抑制作用,如导致对鸡新城疫病毒(NDV)疫苗抗体反应的减弱并使有效抗体滴度持续时间缩短.认识、研究和控制免疫抑制性病毒多重感染,将成为我国禽病界必须面对的一个重要的现实问题.     

鸡传染性贫血病毒与马立克氏病病毒共感染SPF鸡群免疫抑制协同作用的研究

将鸡传染性贫血病毒（Chicken infectious anemia virus,CIAV or CAV）和马立克氏病病毒（Marek s dis-ease virus,MDV）人工单一和共同感染1日龄的SPF鸡,感染后分别于14、21、28、35日龄检测鸡体红细胞压积的变化,并检测鸡群疫苗免疫3周后的抗体反应,以探讨CAV与MDV共感染对鸡体的免疫抑制是否有协同作用。结果表明,在血液分析方面,CAV与MDV共感染组较病毒单一感染组与对照组差异极显著,共感染不仅加重了鸡群贫血现象,而且延长了贫血的病理症状;而在禽流感病毒（Avian influenza virus,AIV）H5/H9疫苗、新城疫病毒（Newcastle disease virus,NDV）疫苗和传染性法氏囊病毒（Infectious bursal disease virus,IBDV）疫苗免疫后3周的抗体检测中,CAV与MDV共感染组较其它各实验组差异极显著,抗体滴度大大低于其它实验组;此外,CAV与MDV共感染组,鸡体生长状况明显差于实验各组,有6只鸡只死亡（6/25）,比病毒单一感染时的死亡率大大增加。综上研究证明,CAV与MDV共感染在免疫抑制作用上有协同作用。     

我国鸡群免疫抑制病感染现状与防控

一、鸡群免疫抑制病的感染现状 1．经对鸡群中禽网状内皮增生病病毒、鸡传染性贫血病毒、禽呼肠孤病毒3种病毒感染的血清学调查发现,绝大多数鸡场中鸡传染性贫血病毒、禽呼肠孤病毒的母源抗体阳性率都非常高,在经过1～3周母源抗体消失后不久,从5周龄开始出现自然感染引发的抗体阳性率逐渐升高;到20周龄时,不论什么遗传品系和地理分布,绝大数鸡群中鸡传染性贫血病毒、禽呼肠孤病毒的抗体阳性率可达到95％～100％。     

鸡传染性贫血病的研究进展

鸡传染性贫血病(CIA)是由鸡传染性贫血病毒(CIAV)引起的一种以雏鸡再生障碍性贫血和全身淋巴组织萎缩为主要特征的病毒性传染病.它和马立克氏病病毒(MDV)、传染性法氏囊病病毒(IBDV)、网状内皮增生症病毒(REV)、禽白血病病毒(ALV)和禽呼肠孤病毒(ARV)是危害养鸡业最常见的免疫抑制性病毒.     

不容忽视的鸡传染性贫血病

在近期流行的鸡病中，大家的注意力大多集中在免疫抑制病、霉菌毒素、新城疫等方面，我们在一些大型养殖公司鸡群的临床中还见到一些值得重视的现象，就是鸡传染性贫血病（CIA）造成的伤害。鸡传染性贫血病毒是由鸡传染性贫血病毒（CIAV）引发的，引起小鸡严重贫血、皮下肌肉出血、骨髓内成红细胞受到破坏，法氏囊、胸腺萎缩。CIAV既可垂直传播，又可水平传播。     

鸡传染性贫血病在我国的流行、诊断及防制

鸡传染性贫血病（Chicken infectious anemia,CIA）最早被称之为贫血综合征、贫血皮炎综合征、出血性贫血病、贫血因子病等,它是由鸡贫血病毒（Chicken infectious anemia Virus ,CIAV）引起的以鸡再生障碍性贫血和淋巴组织萎缩为主要特征的免疫抑制性疾病.CIAV主要侵害肉鸡、后备父母代种鸡和刚开产蛋鸡,可导致雏鸡胸腺、法氏囊及其它淋巴样器官中淋巴细胞严重缺失,并直接降低免疫机能而引起免疫抑制.CIAV与MDV、IBDV共同或先后感染时,或抑制免疫反应提高鸡对这些疾病或其它疾病的易感性,或相互作用可使各自的致病力增强.     

AA肉鸡群中免疫抑制性病毒分子流行病学调查及对疫苗免疫效果影响分析

为了解AA肉种鸡群中免疫抑制性病毒的存在状况及其对NDV疫苗免疫效果和对商品鸡生长的影响,采用多重PCR、RT-PCR和血清学方法对山东某规模化大型AA父母代肉种鸡场的28日龄、180日龄左右种鸡及其所生产的1日龄、30日龄左右商品鸡进行IBDV、MDV、ALV、REV、CIAV及REOV感染状况的调查,并同时对鸡群NDV免疫后抗体水平进行跟踪监测.结果表明种鸡和商品鸡均存在CIAV的感染,并不同程度地存在CIAV与MDV、IBDV等的共感染;发病鸡群NDV抗体水平明显偏低的原因是由于鸡群感染免疫抑制性病毒所致;种鸡群感染CIAV并垂直传播给商品鸡,间接引起商品鸡发生免疫抑制性病毒的共感染,导致商品鸡群30日龄左右生长受阻、免疫能力降低、疾病爆发.     

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。     

In vivo events of retroviral long terminal repeat integration into Marek's disease virus in commercial poultry: detection of chimeric molecules as a marker

The present study demonstrated, for the first time, that not only in vitro, but also in vivo, coinfections with Marek's disease virus (MDV) and each of the three avian retroviruses (reticuloendotheliosis virus [REV], avian lymphoid leukosis virus [ALV], and ALV-J) lead to retroviral long terminal repeat (LTR) integration into MDV. A total of 306 chicken and 59 turkey commercial flocks, submitted for differential avian oncogenic virus diagnosis, served to evaluate the flock mixed virus infection rate, the rate of birds with a multiple virus infection, and the issue of retroviral LTR integration into MDV in vivo. About a quarter of the tumor-bearing commercial flocks carried a mixed MDV and retrovirus infection. A total of 2926 DNA samples were analyzed, including 2428 chicken and 498 turkey DNA samples. Of these, 991 DNAs originated from flocks with a multiple virus infection. In 103 DNA preparations from that group (103/991, 10.4%), including 38 and 56 from chicken blood and tumor tissues, respectively, and nine samples from turkey blood, multiple virus sequences were detected by polymerase chain reaction (PCR). Fifty-six of the 103 samples were further analyzed by the previously developed hot spot-combined (HS-cPCR assay, of which 48% (27/56) contained chimeric MDV and retroviral LTR molecules. When extrapolated to the total samples derived from the flocks with multiple virus infection, that rate implies that about 5% of the DNA samples would carry MDV-retrovirus integration events. Several birds held a variety of chimeric molecules, indicating that several recombination events occurred simultaneously. The validation of the MDV and retroviral LTR chimeric constitution of these molecules was derived by the MDV and retroviral heterologous primers used for their creation by the HS-cPCR assay, Southern blotting and their detection by retroviral LTR probes, and LTR amplification from the gel-purified chimeric molecules. From several molecules, the LTR was sequenced, and a 161-bp retroviral LTR sequence was demonstrated. Our biochemical data imply that a recent integration occurred in the birds. The viability of recombinant viruses represented by the chimeric molecules will be further approached.     

鸡传染性贫血病毒SYBR Green Ⅰ实时荧光定量PCR检测方法的建立

为建立鸡传染性贫血病毒(chicken infectious anemia virus,CIAV)的实时荧光定量PCR检测方法,本试验通过CIAV基因组保守区域设计1对扩增片段大小为180bp的特异性引物,构建pGM-T-CIAV重组质粒,制备阳性标准品,建立SYBR GreenⅠ实时荧光定量PCR标准曲线,并进行敏感性试验、特异性试验和重复性试验。结果显示,CIAV的Ct阈值与标准品浓度在5.33×108至5.33×103拷贝/μL间呈良好的线性关系,相关系数R2=0.998,斜率为-3.443,产物Tm值在86℃左右。该方法与网状内皮组织增生病病毒(REV)、禽白血病病毒(ALV)J亚型、马立克氏病病毒(MDV)、传染性法氏囊病病毒(IBDV)基因组均无交叉反应,敏感性为5.33拷贝/μL,比普通PCR高1 000倍,批内和批间重复试验变异系数均小于3%。结果表明,本试验建立的CIAV SYBR GreenⅠ实时荧光定量PCR检测方法可实现对鸡传染性贫血病的早期诊断及感染程度的定量分析的检测。     

Coinfection of specific-pathogen-free chickens with Marek's disease virus (MDV) and chicken infectious anemia virus: effect of MDV pathotype

Both Marek's disease virus (MDV) and chicken infectious anemia virus (CIAV) infections are prevalent in chickens throughout the world. In the past decade, MDV strains with increased virulence (very virulent plus MDV pathotype [vv+MDV]) have been isolated. The purpose of this experiment was to determine the effects of coinfection of chickens with CIAV and a vv+MDV isolate. Specific-pathogen-free chickens were inoculated at 1 day posthatch with RB1B (very virulent MDV pathotype [vvMDV]) only, 584A (vv+MDV) only, CIAV only, RB1B + CIAV, 584A + CIAV, or nothing. Samples of spleen, thymus, and bursa of Fabricius were collected at 4, 7, 10, and 13 days postinoculation (DPI). Thymic and bursal atrophy at 13 DPI and final mortality at 30 DPI were significantly greater in chickens inoculated with 584A with or without added CIAV, or with RB1B plus CIAV, compared with birds inoculated with RB1B alone. Both amounts of virus reisolated and levels of virus detected by quantitative-competitive polymerase chain reaction were greater at 4 DPI in 584A inoculates compared with RB1B inoculates. To monitor the early cytolytic infection, northern analysis was done with a probe for the MDV immediate early gene ICP4 (infected cell protein 4). In the absence of CIAV, ICP4 expression was more apparent in chickens inoculated with 584A than in those inoculated with RB1B. CIAV coinfection increased ICP4 expression in the spleens of chickens infected with RB1B. These results indicated that inoculation of chickens with the 584A isolate caused a more robust early cytolytic infection compared with inoculation with RB1B alone and support the classification of 584A as a vv+MDV strain. Coinfection with CIAV exacerbated vvMDV strain RB1B infection. The extent of this exacerbation was less evident when birds were coinfected with 584A and CIAV.     

用斑点杂交法同时检测鸡群中的CAV MDV和REV

为研究鸡传染性贫血病毒(CAV)在鸡群中的感染状况以及马立克氏病病毒(MDV)和网状内皮细胞增生病病毒(REV)在鸡群中的发病率，用斑点杂交法对山东省4个肉鸡场和2个肉种鸡场进行CAV、MDV和REV的检测，结果表明除一个肉种鸡场没有检测出REV以外，其他鸡场均同时检测出CAV、MDV和REV，并且发现直接从病科中检测CAV的阳性率(20％)远远低于将病科接种SPF鸡胚后的检出率(80％)。     

商品肉鸡群中MDV、REV、CAV和ARV多重感染的检测

从山东省东营、日照、潍坊、聊城等地区自然发病和临床健康AA商品肉鸡群中分别采集脏器样品,用特异性核酸探针对样品进行马立克氏病病毒（Marek’s disease virus,MDV）、网状内皮组织增生症病毒（Reticuloendotheliosis virus,REV）、鸡传染性贫血病病毒（Chicken anemia virus,CAV）和禽呼肠孤病毒（Avian reoviruses,ARV）检测。结果显示,自然发病AA商品肉鸡群中MDV、REV、CAV和ARV的检出率均较高,分别为69．30％、57．46％、63．60％和67．11％;临床健康AA商品肉鸡群中MDV、REV、CAV的检出率分别为36．96％、43．48％和30．42％,且自然发病和健康鸡群中均存在不同病毒组合的多重感染,感染率分别为85．96％和43．46％。用x2检验进行分析发现,自然发病商品肉鸡群与临床健康商品肉鸡群中MDV、CAV、MDV＋REV、REV＋CAV的检出率和未检出的比例差异极显著（P〈0．01）;REV、MDV＋CAV检出率差异显著（P〈0．05）。对自然发病商品肉鸡的肝脏、脾脏、法氏囊中4种病毒检出率进行X2检验分析发现,MDV在脾脏中检出率显著高于肝脏和法氏囊;REV在法氏囊中检出率显著高于肝脏和脾脏,而CAV和ARV分别在脾脏和肝脏中检出率较高。结果表明,多种免疫抑制性病毒的共感染已普遍存在,是目前AA商品肉鸡易发病且生长缓慢的重要流行病学因素之一。     

我国白羽肉用型鸡群中CAV、REV和REOV感染状况的血清学调查

为了解鸡传染性贫血病毒（CAV）、禽网状内皮增生病病毒（REV）和呼肠孤病毒（REOV）在我国白羽肉用型鸡中的感染状态，在2003—2004年，检测了来自5省市8个公司不同年龄鸡群血清样品中3种病毒抗体的存在状况。结果表明，在送检的75个鸡群中，对CAV、REV和REOV呈现抗体阳性的鸡群分别有64个（85．3％）、36个（48％）和74个（96％）。在总共检测的1764份血清样品中，对这3种病毒的平均抗体阳性率分别为51．4％、9．8％和75．1％。在1日龄雏鸡，对CAV和REOV的平均母源抗体阳性率可达100％和81．1％，但对REV只有7．4％。抗体阳性率随年龄变化的动态分析表明，对REV和REOV的母源抗体在出壳后2～3周内消失，而对CAV的母源抗体则可持续3～4周。对CAV和REOV的抗体从5周龄起再次出现，到20周龄时，所有送检鸡群全部阳性，平均阳性率在90％以上。有近一半送检鸡群对REV呈现抗体阳性，抗体阳性率普遍较低，即使在达到开产年龄后，仍还有很高比例鸡为抗体阴性，即对REV仍为易感鸡。研究表明，我国多数鸡群中都同时存在着这3种病毒的感染，但它们在感染的程度和动态等流行病学特点上显著不同，应根据鸡群中抗体的阳性率分别采取不同的措施。     

Chicken infectious anemia virus infection in Israeli commercial flocks: virus amplification, clinical signs, performance, and antibody status

The impact of chicken infectious anemia virus (CIAV) infection on commercial chicken flocks in Israel was examined by analyzing flocks with or without typical CIAV signs, signs of other diseases, or apparently healthy flocks. In 23 flocks (broilers and layers) of ages up to 8 wk, typical signs of CIAV infection (stunting, gangrenous dermatitis, and secondary bacterial infections) were recorded. When permitted by flock owners, in several cases among these 23 flocks the morbidity, mortality, and performance parameters were recorded; the presence of CIAV was detected by polymerase chain reaction (PCR); and the antibody status of parents and broilers was measured. In addition, total mortality, number of birds sold, total kilograms of meat sold, density (kg/m2), mean age at slaughter, daily growth rate in grams, total kilogram of food consumed, food conversion rate, and the European Index were calculated. We also surveyed flocks affected by other diseases, such as tumors, respiratory diseases, or coccidiosis, and flocks with no apparent clinical signs. The latter flocks were negative by CIAV-PCR, indicating that typical CIAV clinical signs are associated with one-step PCR-CIAV amplification. However, a small amount of CIAV might still be present in these flocks, acting to induce the subclinical effects of CIAV infection. These data indicate a link between the presence of virus sequences and typical CIAV signs and strengthen the concept that CIAV infection has a negative economic impact on the chicken industry.