禽髓细胞性白血病骨髓内瘤细胞核内Ag-NOR蛋白颗粒定量分析

对禽髓细胞性白血病(ML)自然病例和J亚群禽白血病病毒内蒙株(IMC10200)试验感染肉鸡的骨髓组织,采用Ag-NOR染色技术,进行了瘤细胞核仁组成区嗜银蛋白(Ag-NOR蛋白)的形态观察及定量分析。结果表明:随着ML病变加重,瘤细胞核内Ag-NOR蛋白颗粒数量明显增多;其大小、形态均与对照组显著差异。     

IMC10200株ALV-J实验诱发禽骨髓性白血病的研究

对分离自肉种鸡群亚临床感染的J亚群禽白血病病毒IMC1o200株进行了实验感染诱发禽骨髓性白血病的病理学研究.IMC10200株J亚群禽白血病病毒尿囊腔接种11日龄肉鸡胚和SPF蛋鸡胚,孵出后跟踪观察.9周龄时随机抽检感染鸡,感染肉鸡特异引物PCR检测全部为阳性;组织病理学观察感染鸡无明显病变.至21周龄时感染肉鸡出现第一例典型骨髓性白血病病例;感染SPF蛋鸡未见明显J亚群禽白血病相关病变.     

禽白血病髓细胞样瘤细胞的鉴别与误诊分析

J亚型禽白血病病毒(subtype J avian leukemia virus, ALV-J)诱导的髓细胞样瘤细胞在形态上与正常髓系细胞很相似，为明确两者的形态差异和减少对禽髓细胞性白血病的误诊，对ALV-J人工感染病例和自然感染病例中的髓细胞样瘤细胞的形态特征进行了系统观察，并将其与正常鸡髓系细胞进行了鉴别和分析。结果表明，ALV-J感染鸡的血液、肝脏、脾脏和法氏囊等器官中髓细胞样瘤细胞主要是含有粗大嗜酸颗粒的中幼粒细胞和晚幼粒细胞，胞核约占细胞体积的1/2,胞核偏在、未分叶，可见病理核分裂象，其在石蜡组织切片中直径为8～10μm,在血涂片中直径12～14μm;正常幼龄鸡的肝脏、肾脏、法氏囊等器官中也有数量不等的晚幼粒细胞，这些细胞在形态上与髓细胞样瘤细胞很相似，胞浆中含有较多细小的嗜酸颗粒，但细胞体积较小，核浆比小，胞核约占细胞体积的1/3;其在石蜡组织切片中直径为5～8μm,血涂片中未见。研究结果提示，临床中应依据细胞大小、核型以及胞浆成分等特征对髓细胞样瘤细胞进行鉴别，同时应结合病毒核酸检测结果来确诊J亚型禽白血病。     

J亚群禽白血病病毒人工感染肉鸡病理学及组织嗜性研究

本试验从内蒙古某地养鸡场分离出一株J亚群禽白血病病毒(ALV-J),命名为ALV-J IMC10200株.通过人工感染试验建立发病模型,对诱发的肉鸡典型禽骨髓细胞瘤病的病理学变化和病毒的组织嗜性进行研究,为进一步探索本病发病机理、病毒的生物学特性奠定基础.     

禽骨髓细胞瘤病自然病例的病理学研究

对禽骨髓细胞瘤病显性型病鸡、ELISA阳性的稳性型病鸡和阴性对照鸡进行了病理学研究。结果表明，临床症状明显的显性型病鸡肝、脾、肾、睾、丸（或卵巢）、肺等组织有灰白色结节状病灶和不同程度的肿胀，甚至在股骨或肋骨等处出现大小不一的硬固肿块，镜检病变部位有大量髓细胞样瘤细胞，呈典型的骨髓细胞瘤病的病理特征。电镜下瘤细胞病变明显，瘤细胞膜下疑似病毒样粒子；在自然病例肾悬液感染的鸡膛成纤维细胞培养物中巨噬细胞的膜性结构上发现增殖的病毒样凸起。ALV-ELISA抗体检测试剂盒检出阳性、无明显临床症状和眼观病变的隐性型病鸡，光镜下骨髓、肝、卵巢、心肌等组织中可见与显性型相同的髓细胞样瘤细胞，但数量较少。ELISA抗体检测阴性对照鸡骨髓中胞浆含有嗜酸性颗粒的髓细胞明显少于阳性鸡，除在肝、卵巢等组织中偶尔出现外，其他组织很少见到。     

肉鸡禽骨髓细胞白血病的骨髓瘤细胞数量变化规律的研究

为了进一步研究禽骨髓细胞白血病瘤细胞的变化规律,对1日龄肉种鸡人工接种禽骨髓细胞白血病肝组织毒,进行分组试验.分别在2,3,4,5,7,8,23周龄剖杀,观察病变,并取其骨髓、血液涂片瑞氏染色,油镜下观察.对骨髓内瘤细胞数量进行方差分析,t检验,结果表明在2周龄内的瘤细胞差异不显著(P＞0.05),3周龄后瘤细胞开始增殖,数量增多,与对照组差异显著或极显著(P＜0.05或P＜0.01).同时对比同龄肉种鸡其它组织的变化,瘤细胞在5周龄后陆续出现.     

致鸡急性肉瘤组织细胞提取液的泛嗜性研究

为研究致鸡急性肉瘤组织细胞提取液的泛嗜性,本研究采取发生急性肉瘤的山东某地饲养的36日龄817肉杂鸡肉瘤组织细胞提取液接种CEF进行细胞培养,同时人工感染1日龄817肉杂鸡、海兰褐蛋鸡与白羽肉鸡、白来航系SPF鸡等4个品系鸡,结果均复制出同样肉瘤。将复制出的新鲜肉瘤分别制备肉瘤细胞、组织切片与培养12 d的细胞同时采用IFA试验进行病毒鉴定,结果从肉瘤组织中分离到A亚型禽白血病病毒(ALV-A)、J亚型禽白血病病毒(ALV-J)与禽网状内皮增生病病毒(REV)。对1日龄817肉杂鸡进行的致瘤性与接种部位的嗜性研究结果表明肉瘤形成与接种部分密切相关。对不同日龄817肉杂鸡致瘤性结果显示肉瘤发生率对感染日龄有较强的依赖性。本研究表明来自817肉杂鸡的肉瘤组织细胞提取液中存在ALV-A、ALV-J以及REV共感染,具有急性、高致瘤性、泛嗜性等特点,在不同品系、不同日龄鸡均可诱发急性肉瘤。但急性肉瘤由不同病毒诱发还是与多种病毒共感染相关还有待于进一步研究。     

家禽骨髓内红细胞、白细胞发生的显微和亚显微结构特点

利用光镜和电镜对鹅、鸭和鸡骨髓内红细胞和白细胞发生的显微和亚显微结构进行了研究。结果显示 :鹅骨髓内红细胞系的体积比鸭的略大。光镜下鸭、鹅异嗜性粒细胞 ,在早幼阶段胞质内出现少量圆形颗粒 ;在晚幼阶段出现较多暗红色的圆形、杆状、梭形颗粒。嗜酸性粒细胞在早幼阶段胞质内出现少量着桔红色的圆形颗粒 ;晚幼阶段颗粒多呈杆状 ,胞核轮廓清楚。嗜碱性粒细胞在各阶段胞质内散布紫红色的细小颗粒。电镜下鸡原始红细胞多附着在窦壁上 ,核周隙窄 ,核孔数量较多 ;而成熟红细胞多分布在近血窦中央处 ,核周隙宽 ,核孔数量较少。异嗜性颗粒在早幼阶段可分 A型和 B型。嗜碱性颗粒分为 型和 型 ,它们分别在中幼和晚幼阶段出现。嗜酸性颗粒呈均质的圆形     

肉鸡禽骨髓细胞白血病的骨髓瘤细胞数量变化规律的研究

为了进一步研究禽骨髓细胞白血病瘤细胞的变化规律，对1日龄肉种鸡人工接种禽骨髓细胞白血病肝组织毒，进行分组试验。分别在2，3，4，5，7，8，23周龄剖杀，观察病变，并取其骨髓、血液涂片瑞氏染色，油镜下观察。对骨髓内瘤细胞数量进行方差分析，t检验，结果表明：在2周龄内的瘤细胞差异不显著（P＞0.05)，3周龄后瘤细胞开始增殖，数量增多，与对照组差异显著或极显著（P＜0.05或P＜0.01)。同时对比同龄肉种鸡其它组织的变化，瘤细胞在5周龄后陆续出现。     

禽白血病的诊断与防治

<正>禽白血病是一类由禽白血病病毒引起的禽类多种肿瘤性疾病的总称。在自然条件下该类病以淋巴细胞白血病最为常见,其次是成红细胞白血病、成髓细胞白血病、骨髓细胞瘤及内皮瘤等。1临床症状(1)淋巴细胞性白血病。该病是最常见的一种病型,潜伏期长,人工感染1~14日龄的易感雏鸡后,在第14~30周龄发病,14周龄内发病的很少见。自然发病的鸡常于14周龄后出现症状,至性成熟期发病率     

应用组织芯片免疫组化法检测ALV-J

禽白血病J亚群（ALV-J）是英国的Payne和他的同事们在20世纪90年代初从肉鸡中分离出来的新亚群，主要引起肉鸡的骨髓瘤白血病。自1999年，我国一些肉用型种鸡场陆续发生了禽白血病J亚群。并且近几年来，ALV-J已从最初只引起肉种鸡发病开始向蛋鸡及中国地方种鸡蔓延。组织芯片技术是一种新型特殊的生物芯片技术，它能明显提高工作效率，减少实验误差。本研究将组织芯片技术和免疫组化染色结合起来，用特异性抗ALV-J囊膜蛋白gp85的单克隆抗体来检测发病鸡只的各组织器官的组织切片。在肝脏、脾脏、肾脏、卵巢、腺胃、骨髓、髓细胞瘤组织均检出病毒阳性抗原。结果表明组织芯片技术和免疫组化染色相结合为临床诊断ALV-J提供了一个高通量、敏感的检测方法。     

肉鸡骨髓细胞瘤病的PCR诊断和病理学研究

对曾感染ALV-J的某肉种鸡场的5只肉鸡进行了PCR检测和病理学研究。结果：5只鸡中有3例PCR呈阳性，5只鸡的骨髓、肝、脾、肾、肺、十二指肠固有膜等组织中均有不同数量的局灶性或单个散在的髓细胞样瘤细胞，瘤细胞胞浆内含有大量圆球形嗜酸性颗粒，表明该鸡场仍有骨髓细胞瘤病的存在。其中2只鸡还同时有增生性肉芽肿，提示有大肠杆菌的混合感染。     

Lesions of bone and bone marrow in myeloid leukosis occurring naturally in adult broiler breeders

Lesions of bone and bone marrow in myeloid leukosis (ML) occurring naturally in adult broiler breeders were investigated pathologically. During gross examination, nodules and protrusions were commonly observed on the surface of the sternum, ribs, vertebrae, and synsacrum. The bone marrow of all the bones of the body was pale in color. Histologically, granulated myelocytes proliferated in the bone marrow of various bones and in the periosteum of the sternum, ribs, vertebrae, and synsacrum. The first proliferation of tumor cells occurred in the bone marrow of epiphysis. The myelocytes invaded through haversian and Volkmann's canals from the bone marrow to periosteal areas. Hematopoiesis was suppressed by marked proliferation of tumor cells in the bone marrow of the whole bone. Atrophy was also seen in the bones, including medullary bones of the chickens suffering from ML. Proliferation of myelocytes was seen in the bone marrow and periosteum of ossified cartilaginous rings of the trachea and larynx. Marked proliferation of myelocytes was seen in the dura mater of spinal cords, and it subsequently depressed the spinal cords. Bone formation with cartilage was seen in the periosteum of the sternum having marked proliferation of myelocytes in the bone marrow and periosteum. Ultrastructurally, tumor cells showed large nuclei and cytoplasm with large round electron-dense lysosomes. The virus particles were rarely detected in the cytoplasm of tumor cells. The polymerase chain reaction test of tumor samples showed positive for subgroup J avian leukosis virus. This study indicates that the myelocytes can invade through the compact bones to the periosteum in the sternum, ribs, vertebrae, synsarcum, and ossified cartilage of trachea and larynx having thinner compact bones. In addition, the periosteal osteogenesis with cartilage in the sternum may be reactive change against the bone atrophy because of the marked proliferation of myelocytes.     

Lysis of myelocytes in chickens infected with infectious bursal disease virus

In specific-pathogen-free chickens infected with the highly virulent HPS-2 strain or virulent reference GBF-1 strain of infectious bursal disease virus (IBDV), pathologic changes of the bone marrow were investigated. On histologic examination, bone marrow lesions were prominent in the HPS-2 group but only mild in the GBF-1 group. The bone marrow of the HPS-2 group showed severe lysis and depletion of heterophil myelocytes with pyknotic nuclear alteration 2-3 days after inoculation. On examination with an electron microscope, heterophil myelocytes were characterized by shrinkage of the cytoplasm and peripheral condensation of nuclear chromatin. IBDV particles were not detected in altered myelocytes. A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method demonstrated a positive reaction in only heterophil myelocytes. In contrast, nucleosomal DNA fragmentation in HPS-2-infected bone marrow cells was indiscernible by agarose gel electrophoresis. These findings indicate that lysis of bone marrow cells is selectively induced in heterophil myelocytes at an early stage after IBDV infection and independent of virus replication.     

Isolation and some characteristics of a subgroup J-like avian leukosis virus associated with myeloid leukosis in meat-type chickens in the United States.

Several subgroup J-like avian leukosis viruses (ALV-Js) were isolated from broiler breeder (BB) and commercial broiler flocks experiencing myeloid leukosis (ML) at 4 wk of age or older. In all cases, diagnosis of ML was based on the presence of typical gross and microscopic lesions in affected tissues. The isolates were classified as ALV-J by 1) their ability to propagate in chicken embryo fibroblasts (CEF) that are resistant to avian leukosis virus (ALV) subgroups A and E (C/AE) and 2) positive reaction in a polymerase chain reaction with primers specific for ALV-J. The prototype strain of these isolates, an isolate termed ADOL-Hc1, was obtained from an adult BB flock that had a history of ML. The ADOL-Hc1 was isolated and propagated on C/AE CEF and was distinct antigenically from ALV of subgroups A, B, C, D, and E, as determined by virus neutralization tests. Antibody to ADOL-Hc1 neutralized strain HPRS-103, the prototype of ALV-J isolated from meat-type chickens in the United Kingdom, but antibody to HPRS-103 did not neutralize strain ADOL-Hc1. On the basis of both viremia and antibody, prevalence of ALV-J infection in affected flocks was as high as 87%. Viremia in day-old chicks of three different hatches from a BB flock naturally infected with ALV-J varied from 4% to 25%; in two of the three hatches, 100% of chicks that tested negative for virus at hatch had evidence of viremia by 8 wk of age. The data document the isolation of ALV-J from meat-type chickens experiencing ML as young as 4 wk of age. The data also suggest that strain ADOL-Hc1 is antigenically related, but not identical, to strain HPRS-103 and that contact transmission of ALV-J is efficient and can lead to tolerant infection.     

禽白血病病毒J亚群(ALV-J)的攻毒试验

将禽白血病病毒 J亚群 (AL V- J)内蒙株 NM876 1和 NM9996人工接种于 1日龄爱维茵肉种鸡 ,通过眼观、病理组织学检查观察了攻毒鸡的病理学特征 ;通过 PCR和 EL ISA技术检测了病毒感染率、抗体变化规律以及病毒和抗体的相关性。结果显示 ,攻毒鸡从第 3周开始即可检出病毒 ,NM876 1株和 NM9996株病毒感染率分别为 71.4 %和6 4 .3% ;从第 5周开始 ,出现较明显的病理学变化 ,病变特征以骨髓、肝脏、心脏、脾、卵巢等组织内髓细胞增生为主 ,而法氏囊、脑、坐骨神经无变化 ;攻毒鸡抗体消长变化有一定的规律 ,一般在 7～ 8周龄和 18～ 2 0周龄时分别出现 1次高峰 ,而在 4～ 6周龄、10周龄和 2 3周龄分别出现 1次低谷 ,提示鸡场进行 EL ISA检测时要避开这一时期 ;攻毒鸡产生耐受性病毒血症 ,即病毒阳性而抗体阴性 (V A- )的比例较高 (7/14 ,9/14 )。通过以上的研究证明 ,AL V- J内蒙株疾病模型复制成功 ,NM876 1、NM9996可作为原型株进行相关研究     

间接荧光抗体法快速诊断海兰褐蛋鸡J亚群禽白血病的研究

某地区海兰褐蛋鸡发生肿瘤性传染病，经临床症状和病理学检查，在肝、脾、肾、卵巢、输卵管、肺、骨髓、腺胃、肠道等可见胞浆内充满球形嗜酸性颗粒的骨髓瘤细胞，呈灶状或弥漫性的分布。该变化是禽骨髓细胞瘤病病理学的特征性变化，具有证病意义。经用特异性抗J亚群白血病病毒囊膜蛋白gp85的单克隆抗体的免疫组化方法，在病料组织切片上检出病毒抗原，从而确诊为蛋鸡J亚群禽白血病。本研究是尝试用间接荧光抗体法来快速诊断该病。采用特异性抗J亚群禽白血病病毒囊膜蛋白gp85单克隆对组织触片作间接荧光抗体试验，检测J亚群禽白血病病毒抗原。在骨髓、食管、肌肉、输卵管、肾都出现了范围广而且很明亮的荧光；肝、肺、脾的荥光较为明亮；心脏的荧光较弱，但清晰可见。表明在病料组织中存在特异性病毒抗原。     

Heterophil function and resistance to staphylococcal challenge in broiler chickens naturally infected with avian leukosis virus subgroup J

Avian leukosis virus subgroup J has a high tropism for myeloid lineage cells and frequently induces neoplastic transformation of myelocytes. The impact of congenital avian leukosis virus subgroup J infection on the function of circulating heterophils and susceptibility to staphylococcal infection was investigated. Six-week-old broiler chickens negative for exogenous avian leukosis viruses or congenitally infected with avian leukosis virus subgroup J were inoculated intravenously with 10(6) colony-forming units of Staphylococcus aureus, and pre- and postinoculation heterophil function was assessed. All chickens developed a leukocytosis with heterophilia after inoculation, but total leukocyte and heterophil counts were significantly higher in leukosis-negative chickens than in virus-infected chickens. Tenosynovitis was more severe in leukosis-negative chickens, and 2/10 (20%) of the virus-infected chickens had no histologic evidence of tenosynovitis. Osteomyelitis in the tibiotarsus or tarsometatarsus developed in 5/10 (50%) of the chickens in each group. S. aureus was recovered from the hock joint of 6/10 (60%) of the chickens in each group. Heterophils from all chickens exhibited similar phagocytic ability pre- and postinoculation. Heterophils from virus-infected chickens exhibited less bactericidal ability preinoculation than did heterophils from leukosis-negative chickens. However, postinoculation bactericidal ability was similar in both groups. Avian leukosis virus subgroup J provirus was present in heterophils isolated from congenitally infected chickens. Heterophils isolated from broiler chickens congenitally infected with avian leukosis virus subgroup J exhibit no significant functional deficits, and infected and uninfected chickens exhibit similar susceptibility to staphylococcal infection.