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1.
为了解山东省禽腺病毒(fowl adenovirus,FAdV)毒株的基因遗传演化情况及致病性,本试验对山东省两家疑似暴发鸡包涵体肝炎和心包积液综合征鸡场采集的病料(肝脏、脾脏)进行PCR鉴定,并将鉴定为FAdV的2份阳性病料提取病毒液,接种鸡肝癌细胞(LMH)进行毒株传代培养和细胞病变(CPE)观察、PCR检测、TCID50测定、鸡胚致病性试验、SPF鸡回归试验、病毒hexon部分基因扩增及序列分析。结果显示,试验成功分离到2株FAdV,2株分离株细胞传第1代即可观察到CPE,PCR均可扩增出大小为500bp的片段,且2株分离株细胞F5代TCID50分别为10-7.75/0.1mL和10-7.60/0.1mL,均对7日龄SPF鸡胚有强致病性;第1分离株和第2分离株对21日龄SPF鸡攻毒死亡率分别为80%和15%。hexon基因核苷酸同源性分析结果显示,第1分离株与FAdV-4株同源性最高(99.57%),第2分离株与FAdV-8b株同源性最高(99.18%)。这2株FAdV分离株均与Ⅰ群禽腺病毒同源,与火鸡出血性肠炎病毒(HEV)所在的血清Ⅱ群及减蛋综合征病毒(EDSV)所在的血清Ⅲ群禽腺病毒位于不同分支。第1分离株基因型为C型,血清型为4型,命名为FAdV-SDC4株;第2分离株基因型为E型,血清型为8b型,命名为FAdV-SDE8b株。综上所述,FAdV-SDC4和FAdV-SDE8b株属于近几年国内流行毒株。本研究结果可为鸡包涵体肝炎、心包积液综合征的防控工作和疫苗株的选择提供科学依据。  相似文献   

2.
2株Ⅰ群禽腺病毒的分离鉴定及致病性分析   总被引:1,自引:1,他引:0  
为了解山东省禽腺病毒(fowl adenovirus,FAdV)毒株的基因遗传演化情况及致病性,本试验对山东省两家疑似暴发鸡包涵体肝炎和心包积液综合征鸡场采集的病料(肝脏、脾脏)进行PCR鉴定,并将鉴定为FAdV的2份阳性病料提取病毒液,接种鸡肝癌细胞(LMH)进行毒株传代培养和细胞病变(CPE)观察、PCR检测、TCID50测定、鸡胚致病性试验、SPF鸡回归试验、病毒hexon部分基因扩增及序列分析。结果显示,试验成功分离到2株FAdV,2株分离株细胞传第1代即可观察到CPE,PCR均可扩增出大小为500 bp的片段,且2株分离株细胞F5代TCID50分别为10-7.75/0.1 mL和10-7.60/0.1 mL,均对7日龄SPF鸡胚有强致病性;第1分离株和第2分离株对21日龄SPF鸡攻毒死亡率分别为80%和15%。hexon基因核苷酸同源性分析结果显示,第1分离株与FAdV-4株同源性最高(99.57%),第2分离株与FAdV-8b株同源性最高(99.18%)。这2株FAdV分离株均与Ⅰ群禽腺病毒同源,与火鸡出血性肠炎病毒(HEV)所在的血清Ⅱ群及减蛋综合征病毒(EDSV)所在的血清Ⅲ群禽腺病毒位于不同分支。第1分离株基因型为C型,血清型为4型,命名为FAdV-SDC4株;第2分离株基因型为E型,血清型为8b型,命名为FAdV-SDE8b株。综上所述,FAdV-SDC4和FAdV-SDE8b株属于近几年国内流行毒株。本研究结果可为鸡包涵体肝炎、心包积液综合征的防控工作和疫苗株的选择提供科学依据。  相似文献   

3.
研究对临床上疑似禽腺病毒感染鸡病例进行了病毒分离与鉴定。通过对病鸡肝组织研磨液接种鸡胚、电镜观察、PCR及序列鉴定,分离出一株血清4型禽腺病毒,命名为JH。动物回归试验证明,肌肉接种1 000 EID_(50)病毒量对3周龄SPF鸡的致死率高达90%;病死鸡剖解可见典型的心包积液与包涵体肝炎,肝组织切片HE染色可见典型的嗜碱性核内包涵体。这一高致病性血清4型禽腺病毒的分离报道,为探究国内禽腺病毒分子流行与毒力演变提供基础材料。  相似文献   

4.
为了解贵州地区禽腺病毒血清4型(FAdV-4)分离株的致病性和生物学特性,本研究对经PCR检测为FAdV-4核酸阳性的肝脏样本应用鸡肝癌细胞(LMH)进行FAdV-4分离培养,通过观察FAdV-4感染细胞病变,并应用PCR、间接免疫荧光试验(IFA)、电镜等技术对分离病毒进行鉴定;然后通过动物感染试验分析分离病毒的致病性。结果显示,FAdV-4核酸阳性肝脏接种LMH细胞后在72 h出现细胞病变,对LMH细胞具有一定的适应性,盲传5代肝脏样本接种细胞经PCR检测,呈FAdV-4阳性;IFA检测显示FAdV-4感染LMH细胞见有绿色荧光,FAdV-4细胞培养物经电镜观察可见有大量病毒粒子的存在。上述表明,本研究分离获得FAdV-4,将其命名为FAdV-4-GZJS。动物感染试验证实,该病毒能引起鸡胚和雏鸡出现心包积液、肝炎等临床病例所表现的典型症状;荧光定量PCR检测,感染雏鸡肝脏部位FAdV-4含量最高,心包积液次之。FAdV-4贵州株分离鉴定及其生物学特性和致病性的分析,为该病原致病机制研究奠定基础。  相似文献   

5.
对5份不同来源的临床上疑似禽腺病毒感染鸡组织处理后接种6~7日胚龄SPF鸡胚,经过LMH细胞传代,通过血凝性检测和PCR方法对细胞培养物进行鉴定。血凝性检测表明分离物不能凝集鸡红细胞,对Hexon和52K基因的序列分析显示,分离到5株病毒为Ⅰ群4型禽腺病毒(FAdV-4)。动物回归试验结果表明,5个病毒株均能致40日龄SPF鸡死亡,死亡率达80%以上,并表现与FAdV-4感染临床发病相似的症状。研究结果为探究国内FAdV-4分子流行与毒力演变以及病毒感染的预防提供了基础材料。  相似文献   

6.
从山东某疑似感染禽腺病毒的发病鸡群中采集病料并分离到一株禽腺病毒,克隆纯化后进行PCR检测和序列分析,鉴定该毒株为血清4型禽腺病毒,命名为GY株。用LMH细胞繁殖的GY株病毒滴度可以达到107.5 TCID50/0.1mL,并建立了纯净稳定的种子批。动物致病性试验结果显示,攻毒鸡只8/10死亡,剖检均出现心包积液,肝脏肿大、出血或坏死,表明该毒株为强毒株。就攻毒途径、攻毒剂量等方面研究发现,GY株经胸部肌肉注射后发病率和发病时间相对颈部皮下注射途径更有优势,确定攻毒途径为肌肉注射;通过比较不同剂量(5×103.0~5×106.0 TCID50)GY株病毒攻击SPF鸡的致病性,将攻毒剂量定为5×105.0 TCID50。研究表明,GY株可作为禽腺病毒4型攻毒用强毒株。  相似文献   

7.
为进一步了解Ⅰ群禽腺病毒鸭源分离株的致病性和生物学特性,采取接种鸡胚和鸭胚,攻毒SPF鸡和雏鸭,并应用鸡胚成纤维细胞(CEF)和鸡胚肝细胞(CEL)培养观察细胞病变。试验结果表明,SPF鸡胚和鸭胚均出现死亡以及心包积液等典型症状,SPF鸡和雏鸭也引起不同程度死亡,且出现与临床送检病例类似的症状,PCR检测与序列分析证实分离的2株鸭源Ⅰ群禽腺病毒均为FadV-4血清型,病毒接种CEF细胞后没有细胞病变,而接种CEL细胞后则出现很强的聚集性、细胞变圆等典型病变。表明分离的鸭源FadV-4对SPF鸡和雏鸭均具有一定的致病性,对CEL细胞具有一定的适应性。  相似文献   

8.
采集北京市平谷某鸡场病鸡肝组织,经研磨后接种SPF鸡胚,分离到一株禽腺病毒。经PCR扩增,扩增产物测序后进化树分析和病毒中和试验确定该毒株为禽腺病毒4型。动物回归试验复制出相同的临床症状,在电镜下观察到病鸡肝细胞中有典型的腺病毒粒子。综合上述结果确诊病鸡感染了禽腺病毒4型。  相似文献   

9.
2015年8月,河南和湖北某鸡场发生了以心包积水为典型特征的疫情,具有较高的发病死亡率。临床剖检分析为疑似心包积水综合征,病原可能为禽腺病毒。采集心包积水液的病鸡肝脏、心脏等病料接种SPF鸡胚进行病毒分离,同时对病料和接种后的鸡胚尿囊液进行禽腺病毒的PCR检测及序列分析。结果:在病料与尿囊液中均检测出大小约649 bp的片段,将分离到的两株病毒分别命名为HBHP1510和HNXZ1510;序列分析结果表明,两株病毒的同源性为99.6%,与腺病毒4型的同源性最高,为95.1%~97.5%;遗传进化分析表明两株病毒与禽腺病毒4型的亲缘关系最近。本研究表明,此次从心包积水综合征病死鸡中分离的病毒为禽腺病毒Ⅰ亚群4型。  相似文献   

10.
为获得禽腺病毒毒株,将临床疑似Ⅰ群禽腺病毒感染病例的肝组织,通过接种SPF鸡胚进行病毒分离,用PCR方法对分离产物进行扩增,将PCR产物进行序列测定,对测定的序列在NCBI网上进行分析,结果显示,从病料可分离出病毒,用PCR方法特异性地扩增出了与设计片段大小相一致的片段,序列分析表明,分离株序列与禽腺病毒SD1-15分离株序列同源性99%,说明分离株为Ⅰ群禽腺病毒血清4型。  相似文献   

11.
BackgroundInclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH.ObjectivesTo characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes.MethodsSuspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes.ResultsThe hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard.ConclusionsFAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.  相似文献   

12.
Abdel-Alim GA  Saif YM 《Avian diseases》2002,46(4):1001-1006
The pathogenicity of serotype 2 OH strain of infectious bursal disease virus (IBDV) to specific-pathogen-free (SPF) chicken embryos and 2-wk-old SPF chickens and turkey poults was investigated. The virus was pathogenic for chicken embryos after five passages as evidenced by pathologic changes in inoculated embryos. The embryo-adapted virus was not pathogenic for 2-wk-old SPF chickens and turkey poults as indicated by lack of clinical signs, gross or microscopic lesions in the bursa of Fabricius of inoculated birds. Bursa-to-body-weight ratios of the inoculated chickens and turkey poults were not significantly different from those of uninoculated controls. Virus-neutralizing antibodies to serotype 2 IBDV were detected in inoculated chickens and turkeys. Results of this study indicated that the embryo-adapted serotype 2 OH IBDV isolate that is pathogenic for chicken embryos is infectious but not pathogenic in chickens and turkeys.  相似文献   

13.
Y Okuda  M Ono  S Yazawa  Y Imai  I Shibata  S Sato 《Avian diseases》2001,45(4):819-827
The pathogenicity of a serotype 1 fowl adenovirus (FAV-99ZH), isolated from broiler chickens exhibiting gizzard erosion, was investigated in commercial broiler chickens. Five-, 3-, and 1-wk-old commercial broiler chickens were inoculated with FAV-99ZH by both oral and ocular routes. In the 5-wk-old chickens (trial 1), none of which had the maternal antibody to FAV-99ZH, severe gizzard erosions were observed on days 5, 7, and 10 postinoculation (PI). Among the 3-wk-old chickens (trial 2), which were separated into a control group and three treatment groups according to their maternal antibody titer levels, some chickens showed clinical signs such as depression and anorexia. Compared with the control group, all the treatment groups showed decreased weight gain. One treatment group, moreover, showed significantly decreased (P < 0.05) weight gain on day 10 PI. Severe gizzard lesions, such as erosion or ulcers, were observed from day 4 PI in all treatment groups regardless of their maternal antibody levels. The 1-wk-old chickens (trial 3) were separated into a control group and two treatment groups according to their titer levels of the inoculated virus. In spite of high maternal antibody levels, severe gizzard lesions were observed in both treatment groups, which also showed decreased weight gain. One treatment group, inoculated with the higher dose, showed significantly decreased (P < 0.05) weight gain on days 10 and 14 PI compared with the control group. Fowl adenovirus was recovered mainly from gizzard and rectal (including feces) samples from inoculated chickens but was not recovered from liver samples in any of the trials or in any of the control chickens. Although the reproduced disease was similar to that described in a previous report of experimental infection of specific-pathogen-free (SPF) white leghorn chickens with fowl adenovirus, the pathogenicity of FAV-99ZH in commercial broiler chickens was more severe than that in the SPF white leghorn chickens. The results of the present study indicate that FAV-99ZH isolated from gizzard erosion had pathogenicity and produced severe lesions in the gizzards of broiler chickens and that FAV-99ZH could infect and produce illness in broiler chickens with maternal antibodies against this virus.  相似文献   

14.
The pathogenicity of serotype 8 fowl adenovirus (FAV), isolated from gizzard erosions of slaughtered broiler chickens, was investigated. In experiment 1, 29 5-day-old specific-pathogen-free (SPF) chickens were inoculated with the isolates of serotype 8 FAV, M013 (group 1) or G0054 (group 2) strain, via an oral route. There were no clinical signs in any of chickens after inoculation, and mild gizzard erosions were observed macroscopically and microscopically in three inoculated chickens of group 2. FAV was recovered from gizzards and rectums but was not recovered from pancreas and livers from chickens in both inoculated groups. In experiment 2, 27 1-day-old SPF chickens were inoculated with the G0054 strain by intramuscular route. Five, 6, and 3 inoculated chickens died on days 3, 4, and 5 postinoculation (PI), respectively. Four, 3, 1, and 1 inoculated chickens became moribund with severe clinical signs such as ruffled feathers, severe depression and closed eyes from days 3 to 6 PI, respectively. Macroscopically, the common characteristic of the gross lesions of dead chickens and euthanized moribund chickens was discoloration of liver. FAV was recovered from the gizzard, liver, pancreas and rectum. Virus titers in the liver and pancreas were high until day 6 PI. Histologically, necrotizing hepatitis and pancreatitis with intranuclear inclusion bodies were observed in the inoculated chickens. These results indicate that some strains of serotype 8 FAV are able to reproduce not only gizzard erosion by oral inoculation but inclusion body hepatitis (IBH) by intramuscular inoculation.  相似文献   

15.
The pathogenicity of serotype 8 group I avian adenovirus (GIAAV) strains (TR630 and Saga97 strains) from inclusion body hepatitis (IBH) against cyclophosphamide (CY)-treated 3-wk-old specific-pathogen-free (SPF) chickens was examined. SPF chickens were inoculated intramuscularly with 10(7) plaque-forming units of viruses. Both strains from IBH could produce hydropericardium and mortality in CY-treated chickens as hydropericardium syndrome (HPS) that serotype 4 GIAAV strains cause, although they could not induce either hydropericardium or mortality in nontreated chickens. Histologically, hepatocytic necrosis with intranuclear inclusions, pancreatic acinar necrosis with intranuclear inclusions, and epicardial edema were seen in CY-treated chickens inoculated with GIAAV from IBH. Immunohistochemically, these inclusions were positive against GIAAV antigen. There were neither histologic lesions nor positive reactions against GIAAV antigen in nontreated chickens inoculated with GIAAV from IBH. From the present findings, pathogenic characteristics of IBH strains and HPS strains in the chickens were essentially the same.  相似文献   

16.
The mortality and pathology caused by serotype 4 adenovirus, isolated from chickens with hydropericardium syndrome (HPS) in Japan, was investigated in specific-pathogen-free (SPF) chickens. One-day-old to 15-mo-old SPF chickens were inoculated intramuscularly, orally, and intranasally with liver homogenates from HPS chickens or isolated serotype 4 adenovirus. There were no clinical signs before death. The mortality rate in all groups of 1-day-old chicks was 100%, irrespective of the inoculum or inoculation route. Four-week-old chickens inoculated with liver homogenate also had a 100% mortality rate. Five-week-old chickens inoculated with cell culture of HPS adenovirus had a 40% mortality rate. The mortality rates of 7-mo-old hens inoculated with liver homogenates intramuscularly and orally were 75% and 25%, respectively. In 15-mo-old hens inoculated with liver homogenates intramuscularly, the mortality rate was 70%. Gross lesions were hydropericardium and swelling and congestion of the liver with occasional petechial hemorrhages. Histologically, the liver had diffuse or multifocal hepatic necrosis and hemorrhage with intranuclear inclusion bodies noted within hepatocytes. In the spleen, macrophages containing erythrocytes and yellow pigment were prominent in the red pulp. In the lung, a moderate diffuse macrophage infiltration was noted throughout the lung parenchyma, and these macrophages contained yellow pigment. In the pancreas of the chicks inoculated at 1 day old, there was multifocal necrosis of glands with intranuclear inclusion bodies. Intranuclear inclusion bodies were seen also in the gizzard, proventriculus, duodenum, cecum, kidney, and lung of the chicks inoculated at 1 day old. Immunohistochemically, the intranuclear inclusion bodies of various organs showed positive reactions against group I avian adenovirus. Adenovirus was recovered from the liver of chickens with HPS. This study indicates that HPS adenovirus is able to reproduce HPS lesions and mortality in SPF chicks and even adult chickens and that it is a highly pathogenic strain.  相似文献   

17.
Gizzard lesions were formed in specific-pathogen-free (SPF) white leghorn chickens inoculated with fowl adenovirus (FAV). The virus, serotype 1 FAV 99ZH strain (FAV-99ZH), was originally isolated from the gizzard mucosa of commercial broiler chickens exhibiting gizzard erosion with intranuclear inclusion bodies. Five-day-old and 53-day-old SPF white leghorn chickens were inoculated with FAV-99ZH by both oral and ocular routes and then examined at necropsy on days 3, 5, 7, 10, 14, and 21 postinoculation (PI). There were no clinical signs in any of the chickens after the inoculation. Focal gizzard lesions occurred macroscopically, however, in inoculated chickens at several experimental periods. FAV was recovered from tissue samples of the proventriculus, gizzard, pancreas, and rectum by day 10 or 7 PI but was not recovered from liver samples of any of the chickens. These results indicate that FAV isolated from gizzard erosion is able to reproduce gizzard lesions as necrosis and erosion in SPF white leghorn chickens and that it may have a greater degree of tissue tropism in gizzards and other digestive organs than in the liver.  相似文献   

18.
为确诊海南省某鸡场疑似心包积液综合征病例,于2016年5月从海南省某鸡场送检的病鸡中采集病料(肝脏、心脏),研磨,通过卵黄囊接种7日龄SPF鸡胚,并进行鸡胚病变特征观察、禽腺病毒PCR特异性检测、基因测序和Blast序列比对。结果表明,接种鸡胚后5d开始死胚,剖检可见胚体发育受阻,羽毛稀少,体表潮红,鸡胚皮肤表面有出血点;在病料研磨液、尿囊液及胚体肝脏中均检测到大小约为884bp目的片段,将分离病毒命名为HN1605;hexon基因Blast序列比对及遗传进化分析结果表明,分离毒株与2015年山东和湖北报道的Ⅰ亚群腺病毒血清4型(FAV-4)SDSX、SDSX-15、SDNY2-15、HB1510和HBLF-15分离株亲缘关系最近。本研究表明,此次心包积液综合征病原是Ⅰ亚群禽腺病毒血清4型病毒,该病毒依然危害着我国鸡群,疫情不断扩大,并呈现由我国北方地区向南方地区扩散的趋势。  相似文献   

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