鸡传染性支气管炎病毒M41株的致病性及排毒规律

本试验针对鸡传染性支气管炎病毒(IBV)呼吸型M41标准株的S1基因设计并合成引物,构建了阳性质粒标准品;经过不断优化建立了检测IBV-M41 SYBR GreenⅠReal-Time PCR标准曲线。通过对8日龄SPF鸡人工感染IBV-M41发病,采用Real-Time PCR方法跟踪检测其泄殖腔排毒情况;剖检并采集试验鸡的相关器官制作病理切片;在临床观察的基础上记录试验鸡的体质量、体温等指标。结果显示,所建立的检测方法重复性好,特异性高,最低可检测到28.3拷贝/μL的病毒核酸。人工感染病毒3～15 d后在鸡的排泄物中均检测到了病毒;攻毒组鸡临床症状明显,相关组织器官均呈现不同程度的病变。该病毒致病性试验、排毒规律和real-time PCR检测方法的建立,为IBV感染鸡作为实验动物模型进行相关药物的临床药效评价提供方法与依据。     

Kinetics of lymphocytic subsets in chicken tracheal lesions infected with infectious bronchitis virus

The kinetics of T-cells (CD3 positive (+), CD4+ and CD8+ cells) and B-cells (IgG+, IgM+ and IgA+ cells) in chicken trachea were studied immunohistochemically and histopathologically following an intratracheal inoculation of infectious bronchitis virus (IBV). Viral antigen was detected in the cytoplasm of tracheal epithelium from 16 hr to 6 days post-inoculation (p.i.) with a peak on 4 days p.i. A few IgG+, IgM+ and IgA+ cells were detected in the submucosa from 8 hr p.i. Thereafter IgG+ and IgM+ cells were gradually increased in number, and dramatically increased from 3 days p.i., peaked on 4 days p.i., and gradually decreased after 5 days p.i. IgA+ cells were detected in a small number than IgG+ and IgM+ cells during the all experimental period. These B cells mainly existed in the lamina propria, and some cells were recognized in the interepithelial space. After 14 days p.i., small number of IgG+ and IgM+ cells were detected in the germinal center of lymph follicles in the lamina propria. From 24 to 60 hr p.i., a few number of CD3+, CD4+ and CD8+ cells were detected at the perivascular area in the lamina propria. After 3 or 4 days p.i., each positive T-cells increased rapidly in number, and reached on the peak at 5 days p.i. CD3+, CD4+ and CD8+ cells tend to distribute diffusely, perivascular area, and surrounding area of CD4+ cells, respectively. CD4+ cells were dramatically decreased from 7 days p.i., and CD3+ and CD8+ cells were decreased from 14 days p.i. No T-cells were detected in the lymph follicles in the lamina propria.     

鸡传染性支气管炎病毒的组织嗜性研究进展

鸡传染性支气管炎（IB）是鸡的一种急性、高度传染性病毒性传染病。自１９３１年首次报道了以呼吸道病变为主的呼吸道病变型IB犤１犦以来，临床上表现为不同组织嗜性的传染性支气管炎病毒（IBV）毒株，就不断地被分离和鉴定出来，先后出现了嗜生殖道型犤１犦、嗜肾型犤２犦、嗜肠型IBV犤３犦，Gough（１９９２）报道IBV毒株７９３／B能引起肌肉松弛犤４犦，有些IBV毒株混合感染致肿头综合征，以及IBV交叉感染出现的多变异株及重组现象。１９９４年以来，我国的江苏、山东等地流行了一种以腺胃肿胀、腺胃乳头溃疡为特征的传染病，从…     

Induction of chicken interferon by avian infectious bronchitis virus.

The ability of nine strains of avian infectious bronchitis virus (IBV) to induce chicken interferon has been investigated using Semliki Forest virus for the tests. The Beaudette, H120 and Connecticut 46 strains induced interferon in the allantoic fluids of embryonated hens' eggs, the highest titre (1 : 30) being associated with Beaudette; but these as well as the Massachusetts M-41 and H52 strains failed to yield interferon in primary monolayer cultures of chick kidney cells as did all nine strains in organ cultures of chick embryo trachea. None of six strains of IBV investigated was susceptible to the inhibitory effects of chicken interferon.     

鸡传染性支气管炎病毒D971分离株与M41株、T株接种鸡的病理形态学比较研究

鸡传染性支气管炎病毒D971分离株感染SPF鸡后主要表现为消瘦、稀便、伴有轻微的呼吸症状；剖检可见腺胃乳头肿胀、充出血、或乳头凹陷，并附有大量黏液；组织学观察可见腺胃黏膜上皮和固有层有坏死、脱落或溃疡，个别病例见有淋巴样细胞浸润，肌胃黏膜有中度坏死，伴有少量淋巴样细胞浸润，实质器官细胞变性、坏死，脾、胸腺及法氏囊的淋巴细胞有较为明显的坏死。M41株和T株感染SPF鸡后，其腺胃变化不明显。     

鸡传染性支气管炎的免疫机制

由鸡传染性支气管炎病毒(Infectiousbronchitisvirus, IBV) 引起的鸡传染性支气管炎由于病毒血清型较多, 易于发生变异而难以免疫预防,成为养鸡业发展的重大阻力。本文从IBV的分子免疫学结构, 在鸡体内引起的体液免疫和细胞免疫方面进行IB免疫机制分析, 提出了具有借鉴价值的免疫方法。     

Epithelial cell kinetics in the inflammatory process of chicken trachea infected with infectious bronchitis virus

All stages of degeneration and regeneration in chicken tracheal epithelium were studied morphologically following an intratracheal inoculation of infectious bronchitis virus (IBV). Viral antigen was detected in the cytoplasm of tracheal epithelium from 1 to 7 days post-inoculation (d.p.i.) with a peak on 3 d.p.i. At 1 d.p.i., almost all epithelial cells were involved in the degeneration. At this time, labelling index of bromodeoxyuridine (BrdU) in the basal cells showed significantly high value compared with control. At 2 and 3 d.p.i., a great number of basal cells were recognized, but the BrdU labelling index tended to decrease. At 4 and 5 d.p.i., the BrdU labelling index of basal cells significantly decreased than 1 d.p.i., and a few number of regenerated immature ciliated epithelia appeared. At 6 to 11 d.p.i., the ciliated columnar epithelia increased rapidly in number, and returned to the normal appearance except for non-ciliated patch by 13 d.p.i. These results suggested that the tracheal epithelial cells infected with IBV degenerated within 24 hours and proliferating activity of basal cells functioned immediately, and 3 to 4 days later, these basal cells were differentiated to the ciliated epithelia.     

The infection of primary avian tracheal epithelial cells with infectious bronchitis virus

Here we introduce a culture system for the isolation, passaging and amplification of avian tracheal epithelial (ATE) cells. The ATE medium, which contains chicken embryo extract and fetal bovine serum, supports the growth of ciliated cells, goblet cells and basal cells from chicken tracheas on fibronectin- or matrigel-coated dishes. Non-epithelial cells make up less than 10% of the total population. We further show that ATE cells support the replication and spread of infectious bronchitis virus (IBV). Interestingly, immunocytostaining revealed that basal cells are resistant to IBV infection. We also demonstrate that glycosaminoglycan had no effect on infection of the cells by IBV. Taken together, these findings suggest that primary ATE cells provide a novel cell culture system for the amplification of IBV and the in vitro characterization of viral cytopathogenesis.     

肾型传染性支气管炎病毒对鸡嘌呤代谢的影响研究

用T株肾型传染性支气管炎病毒(IBV)人工感染14日龄非免疫土种鸡建立感染模型,于攻毒前、攻毒后48 h、72 h、96 h、120 h 采集血样,测定血浆黄嘌呤氧化酶(xanthine oxidase, XOD)活性,探索鸡嘌呤代谢变化与肾型IBV致病机理之间的关系.结果表明,与攻毒前相比较, 攻毒后48 h和72 h血浆XOD活性明显增强, 差异显著(P<0.01,P<0.05).结果提示,XOD活性升高引起的嘌呤代谢紊乱可能是鸡肾型IB的病理机制之一.     

中药治疗鸡肾型传染性支气管炎

鸡肾型传染性支气管炎是鸡感染传染性支气管炎病毒(肾毒株)所引发的一种急性、高度接触性传染病。其特征是病鸡气喘、咳嗽、打喷嚏、肾炎、下痢、蛋鸡产蛋减少。该病多发于2～4周龄的雏鸡,成年鸡也有发生。由于目前尚无特异药物治疗,给养鸡业造成重大损失。我站门诊部自1994年以来,试用中药治疗本病150群、患鸡126570羽,平均治愈率达93.13%。1　症状鸡群突然出现呼吸困难、伸颈、打喷嚏、流涕、发出气管口罗音。患鸡精神沉郁,羽毛松乱,怕冷打堆,排白色稀粪,饮水量增加。蛋鸡产蛋量下降,常产软壳蛋、畸型蛋及粗壳蛋。死亡率可高达30.00%。2　…     

山东省鸡传染性贫血流行病学调查

本文对山东省12个地市58个鸡群传染性贫血的流行情况进行了调查,共检测了368份血清样品,蛋鸡群、肉鸡群和种鸡群的鸡传染性贫血病毒(CAV)ELISA抗体检出率分别为91.7%、77.6%和84.78%,平均抗体阳性率为83.2%,结果表明近几年山东省鸡群中CAV感染相当普遍,而且蛋鸡和种鸡抗体阳性率明显高于肉鸡群。     

肾病-肾炎型鸡传染性支气管炎的病理学研究

对43只14日龄的SPF鸡人工感染从上海市分离获得的1株肾致病型鸡传染性支气管炎病毒（上海株IBV），分别于接种后第2、7、14、24、28和36d扑杀，对肾、肺等组织器官进行了动态病理学观察。结果显示，感染鸡主要表现呼吸系统和泌尿系统的病理变化。在感染后的第2d，支气管和气囊呈急性炎症变化；第5d肾实质变性坏死，炎性细胞浸润，肺充血；第8～13d，肾和肺的病变最为严重；第14d后，支气管和输尿管转变为慢性炎症，肾出现弥漫性或局灶性间质性肾炎病理变化。将M41标准毒株与上海株IBV的致病力进行比较，发现2个毒株引起的呼吸器官病变基本相同，但M41，毒株对肾的致病力较轻，未见间质性肾炎和输尿管炎病理变化。证实，上海株IBV主要侵害肺和肾，并造成肾实质变性坏死和间质性肾炎变化。     

鸡肾型传染性支气管炎的诊治

鸡肾型传染性支气管炎是由冠状病毒科的传染性支气管炎病毒引起的鸡的一种急性接触性传染病,主要引起鸡水样腹泻和间质性肾炎,造成肾脏损伤,死亡率高,也能造成呼吸道感染,是影响养鸡业的重要传染病之一。2009年6月21日,烟台市福山区某养殖场发生鸡肾型传染性支气管炎。根据疾病流行特点、临床症状、剖检变化及实验室检查,诊断为鸡肾型传染性支气管炎。     

鸡传染性支气管炎的诊断与防制

鸡传染性支气管炎(Avian Infectious Bronchitis,简称IB)是由传染性支气管炎病毒(infectious bron-chitis virus,IBV)引起鸡的一种急性高度接触性传染的病毒性疾病。该病主要侵害鸡的呼吸系统、泌尿生殖系统和消化系统,使病鸡出现呼吸困难、肾脏病变、产蛋下降等症候。雏鸡感染     

Early pathogenesis in chicks of infection with an enterotropic strain of infectious bronchitis virus

One-day-old specific-pathogen-free chicks were inoculated intranasally and intraocularly with infectious bronchitis virus (strain G). At days 1, 3, 5, 7, 10, and 14 postinfection, three birds were euthanatized, and the virus contents of both enteric tissues and some non-enteric tissues were assayed. Immunofluorescence and histopathological studies were also conducted. Six of 30 chicks died of nephritis between days 5-10 postinfection. Gross kidney lesions were the major pathological abnormalities. Inflammation was observed histologically in trachea, kidney, and rectum. High virus titers were found at various times in trachea, kidney, and all enteric tissues except for the jejunum. Relatively high titers of virus were still detectable at day 14 postinfection in the kidney, proventriculus, cecal tonsil, ileum, rectum, and bursa of Fabricius. Immunofluorescence staining showed viral antigens in enterocytes at the tips of villi in the ileum and rectum, and in the bursa. Viral antigens were also demonstrated in the epithelial cells of the trachea and in kidney tubules.     

单抗介导的银加强金标免疫技术检测传染性支气管炎病毒

利用胶体金标记纯化的羊抗鼠IgG,建立了一种银加强金标免疫技术(SECGA)检测IB抗原.检测抗原时先将抗原点样于硝酸纤维素膜(NCM)中央,封闭后浸于1:40 IBV单抗中作用10 min,洗涤后再浸于1:4金标羊抗鼠IgG液中作用1 h,银染10 min后观察,在膜上出现灰黑斑点的为阳性结果,反之,不出现斑点为阴性结果.SECGA对IB抗原最低检测量为0.703 ng/点(2 μL),对含毒尿囊液的最低检测量为102.9 EID50,检测IBV M41在鸡胚中繁殖时以36～54 h含毒量最高;检测SPF鸡气管粘液,至少于感染后3 d内检出病毒.结果表明,SECGA检测抗原可用于IBV早期诊断,具有群特异性、快速、敏感、简便,不需特殊设备、结果易判定等优点,尤适于基层实验室或现场应用.