Pathogenesis of duck plague in the bursa of Fabricius, thymus, and spleen.

White Pekin ducks were inoculated orally with duck plague virus and killed at 24-hour intervals after inoculation. Spleen, thymus, and bursa of Fabricius were collected and examined by light, fluorescent, and electron microscopy. Necrosis of lymphocytes occurred in the bursa of Fabricius, thymus, splenic periarteriolar lymphoid sheath (T lymphocytes), and splenic germinal centers (B lymphocytes). Viral nucleocapsids were present in the karyoplasm of lymphocytes, but these cells necrotized before virions were formed. Periarteriolar reticular sheath cells and sinusoidal lining cells in the spleen, epithelial cells in Hassall's corpuscle of the thymus, epithelial cells between the cortex and medulla of the follicles in the bursa of Fabricius, and macrophages in all 3 tissues contained nucleocapsids in the nuclei and virions in cytoplasmic vacuoles before necrosis occurred.     

Pathological lesions following an experimental intoxication with aflatoxin B1 in broiler chickens

A follow-up of chickens dosed orally over 21 days with 0.2 and 3 micrograms of aflatoxin B1 (AFB1) g-1 of bodyweight daily and their subsequent recovery 10 days after withdrawal of contaminated food was conducted. Vacuolation of liver cells during the initial days of the intoxication and cellular depletion in the follicle medulla of the bursa of Fabricius were the lesions which appeared first and persisted during the recovery phase in both groups of intoxicated animals. The intensity of these lesions and their persistence was related to the dose of aflatoxin ingested. A significant reduction in the bodyweight and absolute weights of liver, bursa of Fabricius spleen and thyroid was observed in the higher dose group.     

Histomoniasis in the bursa of fabricius of chickens

Histomoniasis was diagnosed in a flock of 6-wk-old commercial chickens. Clinical signs included depression, stilted gait, inappetence, and a slight increase in mortality. At necropsy, there were pale-yellow to dark-gray circular and depressed necrotic lesions in the liver. The ceca were enlarged and impacted with caseous cores. Cecal worms were not observed either at necropsy or on histopathology. Histomonads were demonstrated microscopically within the bursa of Fabricius in addition to the liver, ceca, and spleen. This is the first report of the presence of histomonads in the bursa of Fabricius in commercial chickens.     

肉鸽法氏囊、脾脏和胸腺的形态学、组织学观察

[目的]为今后研究肉鸽免疫功能提供其免疫器官的形态学、组织学观察依据。[方法]选取40日龄的肉鸽,摘除法氏囊、脾脏和胸腺进行形态学观察;通过制作常规石蜡切片,苏木精—伊红染色,显微照相进行组织学观察。[结果]40日龄肉鸽法氏囊黏膜上皮完整,黏膜固有层腔上囊小结数量减少,体积变小,中间出现空泡,淋巴细胞减少;法氏囊小结中皮质和髓质界限不明显,小结周围有大量的结缔组织增生,导致法氏囊免疫功能降低。脾脏各组织结构发育趋于完善,细胞排列紧密。胸腺髓质中淋巴细胞较少,可见胸腺小体;皮质中淋巴细胞较多。[结论]40日龄的肉鸽法氏囊开始萎缩,免疫功能降低,而脾脏和胸腺免疫功能正常。     

Anatomical distribution of T- and B-lymphocytes in Marek's disease--an immunohistochemical study

Neural lesions of Marek's disease, Marek's disease tumours in the ovary, liver, and kidney, as well as spleen and bursa of Fabricii of chickens bearing Marek's disease tumorous infiltrations, were examined by a new immunohistochemical technique basing upon Sternbergers unlabelled antibody enzyme method which allows the exact localization of lymphoid cells based on their surface antigens. Type C neural lesions contained T-lymphocytes almost exclusively. Type B neural lesions had relatively high proportions of T- and B-lymphocytes, and severe type A neural lesions possessed one part of heavily labelled T-lymphocytes and a number of cells stained weakly by rabbit-anti-chicken-T-cell-globulin. Tumorous infiltrations had similar characteristics as type A neural lesions. Spleen and interfollicular spaces of bursa of Fabricius were infiltrated by T-lymphocytes.     

ChMDA5通路参与传染性法氏囊病病毒致雏鸡法氏囊损伤的研究

传染性法氏囊病病毒（IBDV）为dsRNA病毒,可造成雏鸡法氏囊损伤进而发生免疫抑制;MDA5是能够特异性识别dsRNA病毒的模式识别受体。为研究鸡MDA5（chMDA5）信号通路在IBDV致雏鸡法氏囊病理损伤中的作用,试验选取50只14日龄SPF雏鸡随机分为IBDV感染组和空白对照组,每组25只,IBDV感染组雏鸡通过点眼、滴鼻感染IBDV JIC7株病毒液,0.6 mL·只^-1,空白对照组雏鸡经相同途径给予相同剂量无菌PBS,感染IBDV后第1、4、7、21及35天采集雏鸡法氏囊。采用qRT-PCR方法检测法氏囊中IBDV载量,chMDA5及chMDA5信号通路衔接蛋白（chIPS-1）、转录因子（chIRF3和chNF-κB）、下游产物细胞因子（chIFN-β、chTNF-α、chIL-1β、chIL-6） mRNA水平变化;间接免疫荧光法检测chMDA5蛋白表达变化,传统病理学方法检查法氏囊病理组织学变化。结果发现,雏鸡感染IBDV后,其法氏囊中chMDA5、chIPS-1、chIRF3、chNF-κB、chIFN-β、chTNF-α、chIL-1β、chIL-6的表达量均显著高于对照组,且法氏囊组织发生形态损伤,上述变化趋势与IBDV载量变化基本一致。结果表明,雏鸡法氏囊chMDA5及其信号转导通路可被IBDV激活,参与到IBDV感染雏鸡法氏囊损伤与抗损伤过程中。     

Comparative pathogenesis of serotype 1 and variant serotype 1 isolates of infectious bursal disease virus and their effect on humoral and cellular immune competence of specific-pathogen-free chickens

Specific-pathogen-free chickens inoculated with isolate VA (variant A) or isolate IM of infectious bursal disease virus (IBDV) were examined for mitogenic response to T-cell mitogens, primary and secondary antibody response to sheep erythrocytes and Brucella abortus, and gross and histologic lesions in thymus and bursa. Both isolates induced comparable depression in the mitogenic and antibody response, and both caused extensive gross and histologic lesions in the bursa of Fabricius. However, bursal necrosis induced by the IM isolate was accompanied by an inflammatory response, whereas the inflammatory component was lacking in the lesion induced by the VA isolate. Furthermore, the IM isolate induced extensive lesions in the thymus, but the VA isolate did not.     

In vitro and in vivo characterization of avian reoviruses. III. Host factors affecting virulence and persistence

Three avian reovirus isolates (2177, 2035, and 1733) were used to determine the effect of the age of chickens at inoculation on virus virulence and persistence. Groups of specific-pathogen-free leghorns were inoculated with three different reovirus isolates of different levels of pathogenicity at 1 day, 1 week, 2 weeks, 3 weeks, or 4 weeks of age. Tissues were examined for the presence of virus and lesions at regular intervals until 8 weeks postinoculation (PI) and then again at 22 weeks PI. Isolate 1733, which is highly pathogenic, was reisolated from the thymus, trachea, liver, intestine, cecal tonsils, bursa of Fabricius, gastrocnemius tendon, and white blood cells. Microscopic lesions were observed in some tissues, including the thymus, liver, spleen, bursa of Fabricius, and gastrocnemius tendons, when sampled within a 7-day period following inoculation. This isolate persisted and produced microscopic lesions in the gastrocnemius tendons for as long as 22 weeks PI. The isolates of intermediate pathogenicity (2035) or low pathogenicity (2177) were isolated less frequently and from fewer tissues than isolate 1733. Isolate 2035 could be found in the gastrocnemius tendons as long as 7 weeks PI, whereas isolate 2177 was never isolated from the tendons, nor did it produce any notable gross or microscopic tissue changes. Birds inoculated at age 1 week or older with any of the three reovirus pathotypes were more resistant to infection than 1-day-old inoculates, as evidenced by a decrease in virus reisolations and a concurrent reduction in the severity of lesions in selected tissues.     

Susceptibility of ducks and duck-origin cell cultures to infectious bursal disease virus

Specific-pathogen-free (SPF) ducks that were 1, 3, 4, 7, 10, 30, and 180 days old were inoculated experimentally orally or nasally with infectious bursal disease virus (IBDV). Attempts to induce clinical disease in ducks with strain J1 or FK-78 of IBDV were unsuccessful. Virus-recovery attempts from organ and intestinal contents were also unsuccessful. No significant gross or histopathological lesions were found in liver, spleen, kidney, heart, or bursa of Fabricius of 1- and 3-day-old ducks at 4 or 7 days postinoculation. The ratios of bursa weight to body weight of 1-, 10-, and 30-day-old inoculated and control ducks revealed no difference at 21 days postinoculation. The ducks responded serologically, however, by developing both virus-neutralizing and agar-gel-precipitin antibodies. Virus multiplied in embryonated duck eggs and duck embryo fibroblast cells but not in duck kidney cells.     

谷氨酰胺对肉鸡免疫器官胚后发育的影响

160只1日龄艾维茵肉仔鸡随机分成4组。分别饲喂添加0％、0．2％、0．4％和0．8％谷氨酰胺（Gln）饲粮28d。每周末取鸡24只，每组6只，颈静脉放血致死，取胸腺、法氏囊和脾脏，称重．制作切片．光镜观察，研究基础日粮中添加Gln对肉鸡免疫器官胚后发育的影响。结果显示：（1）添加0．8％Gln组1、4周龄和添加0．4％Gln组2、4周龄胸腺重量显著高于对照组（P〈0．05），添加0．8％Gln组2周龄极显著高于对照组（P〈0．01）；添加0．4％Gln组1、2、4周龄和添加0．8％Gln组4周龄法氏囊重量显著高于对照组（P〈0．05），添加0．8％Gln组1、2周龄极显著高于对照组（P〈0．01）；试验各组脾脏重量均高于对照组。添加0．8％Gln组4周龄显著高于对照组（P〈0．05）。（2）试验组胸腺小叶皮质增厚，淋巴细胞密集，髓质比例减小。胸腺小体减少、减小。试验组法氏囊皱襞内淋巴滤泡增多，皮质增厚．淋巴细胞增多。试验组脾脏脾小结增多、增大；动脉周围淋巴鞘增厚；椭球增多、增大．细胞排列疏松．鞘毛细血管管腔变大．内皮细胞增高。间隙增大。添加0．8％Gln对肉鸡免疫器官发育影响最为明显。结果表明，日粮中添加Gln对肉鸡中枢淋巴器官胚后发育的促进作用优于外周淋巴器官。并有延缓法氏囊退化作用。本试验从组织学角度证明Gln能够促进机体淋巴细胞的增殖分化及免疫器官的胚后发育，进而提高机体的免疫功能。     

Evidence for bursal involvement in the pathogenesis of hemorrhagic enteritis of turkeys

The pathogenesis of hemorrhagic enteritis in turkey poults infected with hemorrhagic enteritis virus (HEV) at 3 days or at 2 or 5 weeks of age was compared with pathogenesis in poults that had been chemically bursectomized neonatally and exposed to cell-culture-propagated virus at 2 or 5 weeks of age. Conventional poults exposed to HEV at 2 or 5 weeks developed clinical disease, and mortality ranged from 38% to 100%. In addition to the splenic and intestinal lesions usually seen with HEV infection, the pancreas, bursa of Fabricius, and thymus were also affected. In contrast, although they were free from detectable maternal antibody, poults infected with HEV at 3 days of age failed to develop clinical disease or mortality; however, virus was demonstrated by histological and electron microscopic examinations in spleens of these poults. Neonatal chemical bursectomy completely prevented the clinical signs, gross lesions, and mortality induced by HEV in poults at 2 or 5 weeks of age. These findings strongly suggest that an intact bursa is necessary for HEV to induce disease in turkeys.     

Infectious Bursal Disease in New Brunswick

A flock of four week old chickens experienced a disease of sudden onset in which the only symptoms were those of depression shortly before death, and in which the predominant histological lesion was necrosis of lymphocytes in the bursa of Fabricius.

A virus, designated strain Sk-1, was isolated from pooled bursal tissue of affected birds and was serologically identified as a strain of the infectious bursal agent. This virus was chloroform resistant, did not hemagglutinate guinea pig or chicken erythrocytes and did not produce a cytopathic effect in chick embryo tissue cultures. Equivocal results were obtained in filtration studies but the agent was less than 100nm in diameter.

Four week old chicks inoculated with strain Sk-1 developed microscopic lesions in the bursa of Fabricius which were similar to those seen in the original field specimens. Inoculated chick embryos exhibited characteristic macroscopic lesions and necrosis of vascular tissue was a common histological change.

A limited serological survey of local poultry flocks indicated that infection by this agent had occurred in four of the ten flocks examined.

     

Infectious bursal disease virus infection in the quail-chicken hybrid

Hybrids produced from crossing Cornell K-strain white leghorn chickens and Line II Japanese quails were studied for susceptibility to infection with infectious bursal disease virus (IBDV). Quail-chicken hybrids were infected successfully following inoculation with IBDV at 14, 21, or 52 days of age. In most cases, precipitating antibodies were detected in serum by 10 days postinoculation (PI). Although no clinical signs or gross lesions were evident in the bursa of Fabricius of hybrids, histologic changes in the bursa were detected upon microscopic examination using hematoxylin and eosin staining. Chickens were successfully infected also; they had gross and microscopic lesions in the bursa and produced precipitating antibodies. In addition, staining of bursal sections with low concentrations of peroxidase-conjugated concanavalin A revealed a rearrangement of a leukocyte cell type (probably macrophages) in infected chickens and hybrids. Japanese quails were refractory to infection; they showed no bursal changes and did not form precipitating antibodies.     

Atrophy of the primary lymphoid organs of meat pigeons in Italy associated with circoviruslike particles in the bursa of Fabricius

During a survey effected in a meat pigeon slaughterhouse of central Italy, atrophy of primary lymphoid organs (bursa of Fabricius and thymus) and hypoplasia of bone marrow were observed. Histologic, ultrastructural, and hematologic examinations were performed on a total of 80 randomly selected 30-day-old meat pigeons. By histologic studies, lymphocytic depletion and necrosis with cyst formations in the bursa of Fabricius were detected in all subjects that showed thymus and bursa atrophy at necropsy. Basophilic intranuclear inclusions were also observed in bursal cells. After ultrastructural studies, these inclusions were proved to be viral particles resembling circoviruslike particles in morphology and size. Severe lymphocytic depletion of the bursa was plausibly associated with the presence of the viral particles.     

不同来源传染性支气管炎病毒诱导SPF鸡发病的免疫机制研究

试验旨在探讨不同来源的传染性支气管炎病毒(Infectious bronchitis virus,IBV)诱导SPF鸡发病的免疫机制。选用140只1日龄SPF白来航鸡,随机分为4组,3组攻毒组通过滴鼻点眼途径分别接种鸡源IBV强毒株、鸡源IBV弱毒株和野鸡源IBV毒株3个毒株,对照组以同种方式接种等量灭菌的磷酸盐缓冲液。在感染后12 h、36 h、72 h、7 d和14 d,每组随机选取5只进行剖检,并分别采集法氏囊、肾脏和气管组织,剩余鸡用于观察临床症状、发病及死亡情况。应用实时荧光定量PCR检测攻毒后不同时间点采集的各组织中IBV的病毒载量、Toll样受体(Toll-like receptors,TLRs)及部分细胞因子(白细胞介素(interleukin,IL)和干扰素(interferon,IFN))表达量的变化。结果显示,感染不同来源IBV毒株之后仅鸡源IBV强毒株感染组SPF鸡出现抑郁、翅膀下垂、甩头等典型的临床症状,且在感染后5~10 d共有7只死亡,死亡率为20%。病理剖检发现,感染鸡源IBV强毒株的鸡肾脏肿大、尿酸盐沉积和有花斑样病变,而感染野鸡源IBV毒株、鸡源IBV弱毒株和对照组的鸡无明显的眼观病变。实时荧光定量PCR结果显示,在鸡源IBV强毒株组的法氏囊、肾脏和气管3个组织中均检测到病毒。对照组和野鸡源IBV毒株组中均未检测到病毒,鸡源IBV弱毒株组只在部分组织中检测到病毒。在感染后72 h,鸡源IBV强毒株组与其他各组相比,TLR1、TLR2、TLR3、TLR5、TLR7和TLR15基因在法氏囊中的表达量均显著升高(P<0.05),IL-6和IFN-β参与更强烈的抗病毒免疫反应;在感染后7 d,鸡源IBV弱毒株组与其他各组相比,肾脏中TLR2、TLR3、TLR15、TLR21、IL-6和IL-18基因表达量均显著升高(P<0.05)。野鸡源IBV感染后36 h法氏囊组织中IFN-γ基因表达量显著上调(P<0.05)。综上所述,3个IBV毒株中仅鸡源IBV强毒感染引起SPF鸡典型临床发病症状与可视组织病变,且可提高SPF鸡组织中免疫相关因子的基因表达量。本研究结果揭示,不同来源的IBV对SPF鸡的不同致病性与其感染诱导的免疫反应不同有关。     

禽网状内皮组织增生病病毒感染对SPF雏鸡法氏囊细胞Bcl-2和C-myc基因表达及凋亡的影响

【目的】 探究禽网状内皮组织增生病病毒(Reticuloendotheliosis virus,REV)感染禽类后导致免疫器官发生细胞凋亡的机理。【方法】 以1日龄SPF雏鸡为试验对象,将100只SPF雏鸡随机均分为REV感染组和未感染病毒的对照组,REV感染组雏鸡经腹腔感染500 μL REV稀释液,对照组雏鸡经相同途径注射等量灭菌生理盐水,于病毒感染后第1、7、14、21、28和42天,2组雏鸡随机各抽取5只,心脏采血处死雏鸡后快速摘取法氏囊。分别应用HE染色和病理切片成像系统测定分析法氏囊细胞核浆比,TUNEL细胞凋亡原位检测试剂盒测定凋亡细胞数,免疫组化法测定Bcl-2和C-myc阳性细胞数量,实时荧光定量PCR和ELISA法分别检测法氏囊Bcl-2和C-myc 基因mRNA表达和蛋白含量。【结果】 ①REV感染1日龄SPF雏鸡后21~42 d,其法氏囊淋巴细胞凋亡百分比显著或极显著高于对照组雏鸡(P<0.05;P<0.01);②SPF雏鸡感染REV后21和28 d,其法氏囊细胞核浆比显著低于对照组雏鸡(P<0.05);③REV感染SPF雏鸡法氏囊中Bcl-2和C-myc阳性细胞数在病毒感染后21和28 d显著高于对照组雏鸡(P<0.05);④REV感染SPF雏鸡后21 d,其法氏囊Bcl-2和C-myc 基因mRNA表达极显著高于对照组雏鸡(P<0.01)。⑤SPF雏鸡感染REV后,其法氏囊中Bcl-2蛋白含量较对照组雏鸡有不同程度的增加,其中21和28 d分别差异极显著(P<0.01)和显著(P<0.05),病毒感染组雏鸡的C-myc蛋白含量也始终高于对照组雏鸡,且21和28 d极显著增高(P<0.01)。【结论】 REV感染所致SPF雏鸡法氏囊细胞Bcl-2和C-myc的mRNA表达以及蛋白含量异常均与病毒感染导致的法氏囊细胞凋亡密切相关,而法氏囊细胞凋亡数量增加与REV感染引发的机体免疫机能抑制密切相关。     

雏鸭腔上囊的生长及组织发育

通过对1～49日龄雏鸭腔上囊绝对质量和生长指数测定以及组织学观察,探讨雏鸭腔上囊生长及组织发育规律。结果:随日龄增长,腔上囊绝对质量逐渐增高;腔上囊生长指数21日龄达最高;黏膜大皱褶高、宽度、淋巴滤泡面积、皮质宽度均不断增加;小结相关上皮向腔面形成突起,并不断增高增宽。结果表明,雏鸭腔上囊1～14日龄生长较为缓慢,14～35日龄生长较快,35～49日龄发育基本趋于稳定,其中21～28日龄是雏鸭腔上囊生长发育的高峰时期,28日龄时,雏鸭腔上囊组织结构基本发育成熟。     

鸡胚法氏囊的组织发育

本试验通过对孵化至１３－２１日龄的鸡胚法氏囊进行组织学的连续性动态观察,较为详细地描述了早日龄鸡胚法氏囊的组织学分化发育过程。实验结果显示,直至出壳胶鸡胚法氏囊尚未形成较完整的能组织起 体液免疫促进作用的组织基础结构,其完善的形态结构需在后天环境中分化和发育。