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1.
利用鸭源鸡杆菌YU-PDS-RZ-1-SLG分离株制备超声波裂解抗原,建立了可以检测鸭源鸡杆菌多个血清型抗体的间接ELISA方法。包被抗原质量浓度为10mg/L,包被条件为37℃2h,再4℃过夜;封闭液为1%明胶,封闭条件为37℃1h;阴、阳性血清最佳稀释度为1∶100,酶标二抗工作滴度为1∶1 000;底物显色时间为15min。经交叉性试验、阻断试验和重复性试验证实建立的ELISA方法重复性好,特异性强。板内变异系数为2.01%～5.75%,板间变异系数为2.43%～6.20%。间接ELISA方法的灵敏度是微量凝集试验的25～100倍。利用所建立的ELISA方法检测了人工感染鸭源鸡杆菌的4日龄SPF鸡在感染后不同阶段感染组、同居组和空白对照组的血清抗体,并根据感染后不同阶段所测D450值绘制抗体消长曲线,其抗体水平在感染后32～47d开始上升,60d时达到高峰,但维持时间较短,2周后迅速下降。建立的间接ELISA方法可以用于临床病例的血清学快速检测,为进行鸭源鸡杆菌的血清流行病学调查提供了手段。 相似文献
2.
为研究鸭源鸡杆菌(Gallibacterium anatis)在鸡体内的动态分布及鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)对其的影响,将鸭源鸡杆菌和鸡传染性支气管炎病毒分别或同时人工接种SPF蛋鸡,分别于接种后12、24、48、72、96 h对鸡进行剖检,无菌采集鸡的10种组织样品(上腭裂、气管、肺脏、心脏、脾脏、卵巢、肾脏、肝脏、十二指肠和输卵管),利用SYBR Green I定量PCR(Quantitative PCR,qPCR)方法检测鸭源鸡杆菌组、混合感染组在不同时间点、不同器官中的鸭源鸡杆菌DNA含量.结果显示:鸭源鸡杆菌单独感染时,12 h时即可侵袭到气管、心脏、脾脏、卵巢和肾脏,24 h时便可感染肝脏、十二指肠和输卵管,48 h时可传播到肺脏,72 h时感染上腭裂.混合感染组结果表明,鸭源鸡杆菌12h到达卵巢,24 h时即可出现在所有器官.混合感染组各器官的总体qPCR检出率为37.1％,显著高于鸭源鸡杆菌组的25.3％.鸭源鸡杆菌组和混合感染组qPCR检测结果均显示,鸭源鸡杆菌DNA在气管中的检出率最高,在卵巢中达到最高含量.鸭源鸡杆菌能够造成试验鸡的全身感染,该菌主要对鸡的呼吸系统和生殖系统有致病性,气管和卵巢是其主要的致病器官;2种病原同时接种加重了全身感染,IBV促进了鸭源鸡杆菌在鸡体内的传播,尤其促进了其在十二指肠中的增殖,增强了鸭源鸡杆菌对消化系统的致病性,导致鸡腹泻的发生. 相似文献
3.
通过气管、消化道和肌肉注射接种鼻气管鸟杆菌（Ornithobacterium rhinotracheale,ORT）人工感染SPF雏鸡,观察试验鸡的发病情况及症状,并检测各试验组和对照组的体质量;利用建立的PCR检测方法对试验组和对照组的排菌情况进行动态观察;于感染后不同时间随机剖杀试验组和对照组鸡只,采集心脏、肝脏、肺脏和气管等组织固定、切片、H.E.染色,观察各器官病理组织学变化,利用建立的IFA方法对ORT在雏鸡体内的分布进行研究,并进行血常规和血液生化指标检测。结果表明,ORT接种后,主要引起雏鸡的呼吸困难、咳嗽等症状,增重明显降低;病理组织学变化以肝脏、心脏组织细胞变性,气管和肺脏广泛性出血和炎性细胞浸润为特征;IFA阳性信号主要分布于肺脏和气管;从感染后2～8d一直可持续排菌;HGB、WBC、RBC、BUN和AST发生显著变化。这表明ORT感染主要引起雏鸡的呼吸系统病变,气管接种比消化道和肌肉注射接种对雏鸡致病性强,肺脏和气管是ORT侵害的主要靶器官。 相似文献
4.
为明确鸡群经不同途径感染禽网状内皮组织增殖病病毒(REV)的排毒动态,本试验通过设立无特定病原体(SPF)鸡胚6胚龄卵黄囊感染组、1日龄SPF鸡腹腔感染组以及对上述两组SPF鸡分别加入感染鸡的卵黄囊感染同居组和腹腔感染同居组,建立SPF鸡感染REV模型,同时设置空白对照组,在不同日龄记录各组鸡的体重和死亡情况,并综合运用反转录PCR(RT-PCR)、实时荧光定量PCR(RT-qPCR)和间接免疫荧光试验(IFA)3种检测方法对各感染组进行REV的动态监测。结果显示：(1)与空白对照组相比,腹腔感染组和卵黄囊感染组SPF鸡体重增长在感染后第21～70天均受到显著抑制(P<0.05),卵黄囊感染组SPF鸡的死亡率在感染后第49和63天显著升高(P<0.05),腹腔感染组SPF鸡的死亡率在感染后第42天显著升高(P<0.05),卵黄囊感染组SPF鸡血浆中REV病毒载量在感染后第21天显著高于腹腔感染组(P<0.05);(2)卵黄囊感染组SPF鸡自出壳即可检测到病毒血症阳性,并在感染后第21天时达到排毒高峰,腹腔感染组SPF鸡在感染后第14天时达到排毒高峰;(3)卵黄囊... 相似文献
5.
用108ELD50基因Ⅶ型新城疫病毒(NDV)强毒对20只樱桃谷鸭和SPF鸡进行攻毒,同时分别设10只同日龄樱桃谷肉鸭和SPF鸡为空白对照,进行同居感染。试验鸭攻毒后1、4、7和14d取喉头、泄殖腔棉拭和肝、脾、肾等内脏组织进行病毒分离,同时采血进行ND抗体检测。SPF鸡攻毒后5d内100%死亡,对照组9d内100%死亡。试验鸭攻毒组和对照组在14d观察期内无明显临床症状,攻毒后第7天开始部分试验鸭喉头、泄殖腔棉拭和内脏分离到NDV,第14天试验鸭血清NDHI抗体阳性。结果表明NDV强毒感染鸭群后并不引起明显的发病、死亡,但是病毒可以在其体内复制,并通过喉头和泄殖腔排毒,具有重要的流行病学意义。 相似文献
6.
本研究拟通过禽网状内皮增生病病毒(REV)分子克隆化病毒REV-C99株对1、8日龄SPF鸡的致病性比较,探讨REV的致病性与感染日龄的关系。在1、8日龄SPF鸡,分别腹腔注射REV-C99株1000个TCID50.只-1,以新城疫病毒(NDV)灭活疫苗免疫后HI抗体滴度的测定结果为指标,比较了REV对不同日龄SPF鸡的致病性。结果表明,与对照组相比,1日龄SPF鸡感染REV后,即使经过NDV灭活疫苗的二次、三次免疫,对NDV的HI抗体滴度仍然差异显著(P0.05);而8日龄SPF鸡感染REV后则随着感染时间的延长以及NDV灭活疫苗的二次、三次免疫,对NDV的HI抗体滴度差异不显著(P0.05)。本研究表明,REV感染1日龄SPF鸡可显著抑制对NDV灭活疫苗免疫后的体液免疫反应,并且这种免疫抑制作用可持续至4个月;而8日龄SPF鸡感染REV后更多表现为一过性的致病作用。显然,REV的致病性对雏鸡感染日龄具有很强的依赖性,从而揭示出控制REV早期感染的重要性并具有现实意义。 相似文献
7.
雏鸡是鸡传染性贫血病毒（Chickenanemiavirus,CAV）的最易感宿主。为系统了解中国国内CAV海安分离株（HA9805）感染雏鸡后的病原动态及与血清学和致病性的关系,本研究对1日龄SPF鸡进行了人工感染实验,感染后不同时间进行样品采集,并用病毒学、血清学以及组织病理学方法进行了检测与分析。研究结果发现,CAV海安分离株（HA9805）通过胸肌接种可引发雏鸡CAV病毒血症,引起CAV特征性临床症状及病理变化,并导致雏鸡高达13．3％的死亡率;血清中CAV核酸以及中和抗体的产生与组织病理变化以及疾病进程密切相关。该结果为CAV的早期诊断,制定有效的预防与控制措施提供了科学依据。 相似文献
8.
为研究不同禽源贝氏隐孢子虫对雏鸡的致病性差异,人工感染3日龄雏鸡鸡源、鸭源、鹌鹑源、白文鸟源贝氏隐孢子虫.结果发现,各感染组雏鸡潜隐期均为4d,显露期为11～18 d,均未发生死亡,但其增重均受影响.鸡源C.baileyi(林州株)、C.baileyi(郑州株)和鸭源C.bailey(郑州株)感染组雏鸡临床症状较为明显,发病率较高,分别为71.4％、85.7％和77.1％,剖检可见气管有较多粘液分泌物,法氏囊粘膜充血肿胀,平均增重较不感染对照组分别减少34.5、53.1和19.7 g；鹌鹑源C.bailey(武陟株)和白文鸟源C.bailey(郑州株)感染组雏鸡仅部分表现轻微临床症状,发病率分别为31.4％和28.6％,平均增重较不感染对照组分别减少39.7和43.1g.结果表明不同禽源C.bailey对雏鸡均有不同程度的致病性,以鸡源和鸭源C.baileyi致病性较强. 相似文献
9.
7日龄 SPF鸡经滴鼻、点眼感染 PMV- 2 ,可引起轻微的呼吸道症状 ,病理组织学观察可见气管黏液分泌亢进和轻微的淋巴细胞浸润 ;感染 PMV- 2后的 1天 ,2天 ,3天… 11天 ,定位采取气管、肺、肝、脾、肾、心、大脑、法氏囊、盲肠扁桃体等组织 ,检查病毒的分布规律 ,结果表明 ,PMV- 2在法氏囊、气管、肺、胸腺、脾、肾、大脑中均有分布 ;SPF雏鸡感染 PMV - 2后 ,接种鸡新城疫克隆 30疫苗 ,免疫后 5 - 30天 ,每 5天 1次检测血清中 ND的 HI抗体效价 ,结果表明 ,PMV- 2感染组比对照组的 HI抗体效价平均低 2 log2 ,统计结果显示差异极显著。雏鸡感染 PMV - 2后对新城疫疫苗的免疫应答有影响 ,从而可能抑制机体的免疫功能 ,危害养鸡生产。 相似文献
10.
为研究鸡源网状内皮组织增殖病病毒(REV)对HBK无特定病原体(SPF)鸭的致病性,利用鸡源REV现地分离株REV-HN,分别接种鸭胚成纤维细胞(DEF)、鸭胚(尿囊腔和卵黄囊腔接种)和雏鸭(口腔接种),试验设空白对照组,雏鸭还设有同居感染组.接种REV后不同时间收获DEF和胚体,及雏鸭的外周血淋巴细胞(PBL)和免疫器官,提取基因组DNA,通过RT-PCR和PCR方法检测REV前病毒env基因.结果在DEF中未检测到特异性的env基因片段,而在卵黄囊接种后72 h鸭胚胚体和试验感染的雏鸭PBL中检测到.接种后30 d,在1羽感染REV的雏鸭肾、胸腺和法氏囊中检测到REV前病毒.本研究为利用SPF鸭研究REV提供了依据. 相似文献
11.
Over the past 3 years the frequency of Salmonella hadar infections has increased in Belgium in both poultry and humans. Therefore, the course of infection with S. hadar in poultry was investigated. One day-old and 4 week-old specific pathogen-free chickens were orally infected with one of two S. hadar strains, SH1 or SH2. Mortality was 6% (SH1) and 17% (SH2) in birds infected at 1 day-old. Chickens infected at 1 day-old with SH2 showed a mild diarrhoea. The S. hadar faecal excretion in birds infected at 1 day-old remained high throughout the experiment until 12 weeks post-inoculation (pi). Faecal excretion was lower in older birds. Antibodies to S. hadar were observed from 2 weeks pi (SH2, infected at 1 day-old) or 4 weeks pi (SH1, both groups; SH2, chickens infected at 4 weeks of age). The percentage of chickens with antibodies was higher after infection at 1 day-old than after infection at 4 weeks of age. In a second experiment 1 day-old chicks were infected with SH1 and autopsied at regular intervals until 42 days pi. SH1 was isolated from the caeca from 3 h pi onwards and from the liver and spleen from 18 h until 14 days pi. Serous typhlitis and omphalitis were the main lesions. The number of macrophages in the lamina propria of the caecal tonsils was slightly increased from 18 h until 2 weeks pi. In the liver, inflammation was observed in the portal triads and in the sinusoids. This study indicates that infections with S. hadar lead to intense colonisation of the gut and extensive faecal shedding. It may also cause invasive infections in 1 day-old chickens. 相似文献
12.
J A Lin H Kodama C Itakura M Onuma T Mikami 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(2):329-337
The new cloned serotype 2 Marek's disease viruses (MDV) of ML-6, ML-9, and ML-22 strains were inoculated in specific-pathogen-free (SPF) chicks to evaluate the pathogenicity and protective efficacy. Chicks inoculated or contact-infected with ML strains showed no gross and histological lesions in lymphoid organs, sciatic plexuses and other visceral organs during 10 weeks of observation periods, indicating that the viruses were non-pathogenic. Moreover, the viruses were found to be spread horizontally among chicks by demonstrating the presence of viremia in contacted chicks at 2 weeks-old. Chicks vaccinated with ML-6 at one day-old were protected against subsequent challenge by inoculation with virulent MDV strain of Md/5 at 4 or 7 days old or by contact infection at 7 days old with chickens previously inoculated with the same strain. 相似文献
13.
Poultry represents an important source of Salmonella infection in man. Despite intensive research on immunity, little is known about the involvement of T cell sub-populations in the immunological response of chickens against infection with non-host-adapted Salmonella (S.) serovars. In this study, the T cell composition of blood lymphocytes (CD4(+)CD8(+); CD4(+)CD8(-); CD4(-)CD8(+); CD8(+)TcR1(+); CD8(-)TcR1(+), CD8(+)TcR1(-)) after oral administration of the non-attenuated S. typhimurium wild-type strain 421 (infection) or the attenuated vaccine strain Salmonella vac((R)) T (immunization) to day-old chicks was investigated and compared with non-treated chickens by flow cytofluorometry. Additionally, the occurrence of T cell sub-populations (CD4(+); CD8(+); TcR1(+)(gammadelta); TcR2(+)(alphabeta(1))) in ceca, spleen and bursa of Fabricius of the birds was studied immunohistologically. Blood samples and tissues were examined between days 1 and 12 of age.Chicks inoculated with S. typhimurium 421 or Salmonella vac((R)) T showed significantly elevated percentages of CD8(+)TcR1(+) in blood on days 7, 8 and 9, or on day 8 in comparison to control animals. The CD4 to CD8 cell ratio was about 3:1 in infected animals on day 5 of age. In the organs of treated chicks the numbers of CD8(+)(gammadelta) and TcR1(+)(gammadelta) cells had markedly increased on days 4 and 5 in ceca, 8 and 9 in the bursa and 9 and 12 in the spleen. Moreover, infected or vaccinated birds revealed larger quantities of CD4(+) and TcR2(+) T cells in ceca on days 4 and 5. As shown by double staining, the TcR1(+) cells in the organs of infected animals additionally carried the CD8 antigen.In conclusion, immunization of day-old chicks with the attenuated Salmonella live vaccine strain resulted in the same changes in T cell composition as seen after infection with the non-attenuated Salmonella wild-type strain, but at a lower level. The remarkable increase of CD8(+)TcR1(+)(gammadelta) double positive cells in treated birds indicates an important role of this cell sub-population in the immunological defense of chickens against Salmonella exposure. 相似文献
14.
本研究从2008山东表现神经症状的某新城疫疫苗免疫鸡群中分离到一株病毒。通过鸡红细胞凝集试验和血凝抑制试验,表明为新城疫病毒,将其命名为ck/CH/LSD/08-29。根据GenBank上登录的新城疫病毒基因组序列,设计14对引物经过RT-PCR扩增病毒感染的尿囊液,同时应用3'-RACE和5'-RACE技术,扩增得到该病毒全部基因组序列。结果表明,该毒株基因组由15186个核苷酸序列组成,编码6种结构蛋白,在RNA上的顺序依次为3'-NP-P-M-F-HN-L-5'。F基因裂解位点处的氨基酸序列为112R-R-Q-K-R-F117,符合强毒株氨基酸序列特点。通过F基因裂解位点附近高变区389个核苷酸序列构建遗传进化树,证明该毒株在分类地位上属于基因Ⅱ型。此外,病毒感染SPF鸡试验证明ck/CH/LSD/08-29毒株属于强毒嗜神经型毒株,能引起SPF鸡高致死率。 相似文献
15.
The aim of the study was to analyse experimental transmission of Mycoplasma synoviae, an avian pathogen. Three experiments using specific pathogen-free day-old chicks placed in isolators were conducted. In the first experiment, the birds were introduced in an isolator previously contaminated with a M. synoviae broth culture. After 34 days, these birds were eliminated and, for the second trial, the chicks were introduced in the same isolator without disinfecting. In the third assay, the chicks were placed in an isolator containing a mixture of food, feathers and dust collected less than an hour earlier from a M. synoviae infected laying hen flock. In the second and third experiments in order to exacerbate the M. synoviae infection, the birds were inoculated with infectious bronchitis (IB) virus. The presence of M. synoviae in the environment and in tracheal swabs was monitored by culture, a multiplex PCR (mPCR) detecting M. synoviae and Mycoplasma 16S rDNA and a multiplex RT-PCR (mRT-PCR) detecting the M. synoviae mRNA coding for a membrane protein and Mycoplasma 16S rRNA. In in vitro experimental conditions, M. synoviae mRNA and 16S rRNA were detected up to 20 min and 23 h respectively after mycoplasma death. In the first assay, the first infected bird was detected on the 13th day. In the second trial, culturable M. synoviae or viable M. synoviae were detected in the isolator for 3 or 4 to 5 days respectively after depopulation of the birds of the first assay whereas the first culture positive tracheal swabs were detected on the 33rd day, after IB inoculation. In the third experiment, the first infected birds were detected on the 54th day. Thus, the different assays showed that M. synoviae contaminated material (dust, feathers and food) can infect chicks, sometimes after remarkably long silent periods. 相似文献
16.
采用间接ELISA法检测雏鸡初次及二次感染毒害艾美耳球虫(Eimerianecatrix)后血清免疫球蛋白含量的动态变化。结果表明,雏鸡初次感染E.necatrix 后10 d~ 14 d血清IgG, IgM, IgA 含量开始增加,16 d~18 d达到峰值;雏鸡二次攻击性感染E.necatrix 后2 d~7 d,其血清的上述3种免疫球蛋白含量均不同程度低于初次感染雏鸡,随后开始回升,至10 d~14 d明显高于相应对照及初次感染雏鸡。血清抗体,特别是IgG介导的体液免疫,在雏鸡抵抗E.necatrix初次及二次感染中发挥了重要作用。 相似文献
17.
Histopathologic findings in chickens experimentally infected with Gallibacterium anatis by nasal instillation 总被引：1，自引：0，他引：1
Zepeda VA Calderón-Apodaca NL Paasch ML Martín PG Paredes DA Ramírez-Apolinar S Soriano-Vargas E 《Avian diseases》2010,54(4):1306-1309
Histologic findings in chickens experimentally infected by nasal instillation with reference strains of Gallibacterium anatis are described. No clinical signs were observed in experimentally infected birds; however, sequential histologic examinations of trachea, lung, air sacs, and liver revealed lesions in all infected chickens. Our observations suggest that the reference strains of G. anatis used in this experiment are capable of causing primary infection in chickens. Despite that the experimental birds were inoculated by intranasal route, lesions were detected in the liver, suggesting a probable bacteremia. Because several degrees of severity were established in histopathologic lesions, probable variations in virulence, among the experimental strains, also are discussed. 相似文献
18.
The effects of concurrent cage contamination with Salmonella typhimurium and Eimeria tenella on the establishment of salmonella infection in day-old chickens were investigated. Chickens were divided into five groups: uninfected recipient birds placed in a cage contaminated by donor birds infected with E. tenella and S. typhimurium; E. tenella-infected recipients placed in a cage contaminated by S. typhimurium-infected donors; uninfected recipients placed in a cage contaminated by S. typhimurium-infected donors; E. tenella-infected recipients placed in a cage contaminated by uninfected donors; and uninfected recipients placed in a cage contaminated by uninfected donors. Three identical trials were conducted. Recipient birds were necropsied 4, 7, and 11 days after caging. In the cage where donor birds infected with both organisms had been reared, S. typhimurium counts in feces and number of feces positive for S. typhimurium were significantly (P less than 0.05) greater than those in the other cages on days 0, 4, and 7 after caging. Moreover, in this cage, more chicks died, counts of S. typhimurium in cecal contents were greater, and more birds were positive for S. typhimurium than in the other groups. This suggests that S. typhimurium infection in day-old chickens is enhanced in cages contaminated with E. tenella and S. typhimurium compared with infection in cages contaminated with S. typhimurium alone. 相似文献