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1.
从山东省多个地区的商品肉鸡养殖场采集64份疑似病料,通过RT—PCR方法检测出37份禽偏肺病毒阳性病料,以此作为病毒分离材料接种SPF鸡胚,盲传至第7代时鸡胚生长明显迟滞。取阳性尿囊液在CEF中培养,呈现禽偏肺病毒典型细胞病变(CPE):细胞变圆、悬浮,有合胞体形成。将分离株接种于Veto细胞及DF-1细胞均出现类似病变。对分离株F基因进行测序,并与GenBank中发表的部分代表序列比较。结果显示,与B亚型禽偏肺病毒的核苷酸同源性最高,为97.4%~99.3%;与A亚型禽偏肺病毒核苷酸同源性较低,为77.4%~78.1%;而与C亚型禽偏肺病毒核苷酸同源性最低,为69.5%~69.7%。基因分型显示分离株为B亚型禽偏肺病毒,将该毒株命名为禽偏肺病毒SDWF株。  相似文献   

2.
以简便技术制备鸡胚皮肤(CES)细胞,进而进行了鸡马立克氏病(MD)SB-1株病毒适应于CES细胞和致CES细胞病变效应(CPE)的研究.试验表明,在形成CPE的细胞中可检出核内包涵体,MD的SB-1株病毒的CES细胞毒的蚀斑形成单位(pfu)为105.7201-7.2122/m L.本研究为以后用CES细胞增殖SB-1株病毒以代替鸡胚成纤维细胞(CEF)制造疫苗提供了依据.  相似文献   

3.
We investigated the interaction between Newcastle disease virus (NDV) and Escherichia coli in cell cultures, embryonated eggs, and 8-wk-old chickens. We measured the interactions on the basis of bacterial adherence and NDV hemagglutination titer in chickens, chicken embryos, and chicken embryo cell culture. Depending on the inoculation order of E. coli, a significant alteration of the growth of NDV was observed in both chickens and chicken embryos. When certain strains of E. coli were given before NDV exposure, the virus titers were lowered. In chickens, the mean virus titer was significantly (P < 0.05) lowered in the crop, the proventriculus, the gizzard, and the jejunum. However, there were no significant differences (P < 0.05) between the two groups for NDV titers in the duodenum, ileum, and cecum. In chicken embryos, when E. coli serotypes O78 and O119:B14 were inoculated before NDV exposure, the mean NDV titers were significantly (P < 0.5) lowered. However, there were no significant differences (P < 0.05) in NDV titer between the two groups when E. coli serotypes O78:K80:NM and O1ab:K NM were inoculated 24 hr before NDV exposure. When NDV was given prior to E. coli exposure, NDV titer was higher in both chickens and chicken embryos. In chickens, when NDV was given 48 hr before E. coli inoculation, NDV was detected in the proventriculus, gizzard, jejunum, ileum, and cecum, whereas no virus was detected in the control groups (NDV only). In the crop, NDV was detected at a significantly (P < 0.05) higher titer in the E. coli-inoculated group when compared with the control group that received NDV alone. In chicken embryos, virus titer was significantly (P < 0.05) higher when NDV was given 24 hr before E. coli inoculation for all three NDV strains used (Ulster and V4 strains). Adherence of E. coli to chicken embryo kidney (CEK) cells was significantly higher (P < 0.05) when the CEK cells were infected first with NDV and then by E. coli. The mean bacterial count per microscopic field in NDV-uninfected monolayers was eight compared with 112 for the NDV-infected monolayers. In approximately 10% of the fields in NDV-infected monolayers, the bacteria were too numerous to count.  相似文献   

4.
Two avian and seven mammalian cell lines were evaluated for their application in propagating avian reovirus WVU 2937. Cultures were compared for monolayer-formation time, support of viral replication, passages and postinfection time required for expression of cytopathic effect (CPE), type of CPE, and virus yield. CPE was observed on the first passage with infected egg yolk in primary chicken embryo kidney cells, primary through tertiary chicken embryo liver (CEL) cells, and African green monkey kidney (VERO) cells; on the third blind passage of infected supernatant in Georgia bovine kidney cells, Crandall feline kidney cells, and baby hamster cells; on the fifth blind passage in rabbit kidney cells; and on the tenth blind passage in porcine kidney cells. CPE was not observed after 10 viral passages in rabbit bone-marrow cells. Monolayer formation time and postinfection time for CPE expression occurred sooner, and virus yield was greater, with CEL and VERO cells than with other cell lines.  相似文献   

5.
不同新城疫病毒F48E8、N-79、LaSota和分离株X以1∶10000稀释接种9日龄SPF鸡胚,每胚0.2ml,结果发现,F48E8和分离株X毒力较强,可在50小时左右致死鸡胚,LaSota和N-79较弱。F48E8和X株接种鸡胚的肌肉、肝脏、脑和尿囊液均有大量病毒存在,而N-79和LaSota只在尿囊液中有大量病毒存在,胚体病毒极少。  相似文献   

6.
To know growth profiles of canine distemper virus (CDV) on Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero-DogSLAMtag; Vero-DST cells), the propagation of three strains of CDV was tested in Vero-DST cells in comparison with parental Vero cells. Strain MD77 could grow well in both cell lines, but demonstrated no syncytium formation or indistinguishable rounding cytopathic effects (CPE) in Vero cells. Strains Onderstepoort and KDK-1 also grew well in Vero-DST cells with apparent syncytium CPE, while they grew less or no efficiently, respectively, in Vero cells. All three CDV strains demonstrated the peak titers, in Vero-DST cells before reaching to an extensive CPE and drastic decrease of titers at/after full CPE. Immunohistochemistry revealed that viral antigens of all CDV strains were found exclusively in the syncytia in Vero-DST cells, while in Vero cells, viral antigen was identified in their single cells for strain MD77 but none for other strains. Thus, every strain of CDV could grow well in Vero-DST cells and behaved differently against Vero cells. These results would be of practical value for workers of CDV because 1) In Vero-DST cells, by observation of distinct syncytium CPE, the highest titer or the best growth of virus could be identified; 2) In Vero cells, various CDV strains could be readily classified after propagation in Vero-DST cells.  相似文献   

7.
Five virus strains with cytopathogenic properties for the cell cultures of chick embryo fibroblasts and embryo kidney cells were isolated from chickens clinically suspected of suffering from infectious bursitis. According to the form of cytopathic effect (CPE) on cell cultures, according to chloroform and thermal resistance, nature of nucleic acid (RNA), according to the absence of the production of intracellular inclusions, negative haemagglutination, and according to antigenic identity with the reference strain, the isolates were deemed to belong to the viruses of infectious bursitis. This conclusion was also corroborated by histological and serological studies in isolates of infected experimental animals.  相似文献   

8.
9.
Five continuous cell lines, swine testicular (ST), human rectal tumor (HRT 18), fetal rhesus monkey kidney (MA104), bovine turbinate (BT), and quail tracheal (QT35), were evaluated and compared with chicken embryo fibroblasts (CEFs) for their ability to propagate B1 or Texas GB strains of Newcastle disease virus (NDV). The NDV Texas GB strain replicated in all the continuous cell lines used in this study. Only the ST and QT35 cells produced a cytopathic effect (CPE) similar to that produced in CEFs. However, the ST cell line remained attached while displaying CPE, whereas infected QT35 cells detached, as did the CEFs. The B1 strain of NDV replicated in ST cells, MA104 cells, and CEFs but with less CPE as compared with the Texas GB strain. Pretreatment with trypsin did not enhance CPE with either NDV strain at the level tested. Sera evaluated for neutralizing antibody titers to NDV were significantly higher in titer when the ST cell line was used and compared with CEFs. A high correlation was found between the microscopic examination and the tetrazolium dye (MTT) microassay methods for determining the viral neutralization endpoint, thus suggesting the ST cell line and MTT microassay could be used as an alternative to CEFs and microscopic examination for evaluating neutralizing antibodies titers to NDV.  相似文献   

10.
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.  相似文献   

11.
Bluetongue virus was isolated from a sentinel herd in British Columbia. Virus isolation was by intravenous inoculation of embryonated chicken eggs and subculture in BHK-21 cells. The cytopathic agent was identified as bluetongue virus by electron microscopy and the immunoperoxidase test. The serotype was identified as serotype 11 by virus neutralization.  相似文献   

12.
Five isolates of infectious laryngotracheitis virus were compared by pock formation on the chorioallantoic membrane of embryonated eggs, plaque size in chicken embryo kidney tissue culture, and antigenic relationship using reciprocal kinetics of neutralization. The A4557-5 strain of infectious laryngotracheitis virus, which causes mild respiratory disease, produced pocks with a zone of edema on the chorioallantoic membrane. A virulent virus (Virus 1), isolated from an outbreak of severe disease characterized by a diphtheritic laryngotracheitis, produced the largest plaques in chicken embryo kidney cell culture. Other virulent viruses (Viruses 2, 3 and V154) did not have unique growth characteristics when grown on the chorioallantoic membrane or in chicken embryo kidney cell culture. All viruses were closely related antigenically as shown by kinetics of neutralization but viruses 2 and 3 were not homogeneous with the other three viruses when neutralized by anti-V154 chicken serum. Following aerosol infection, chickens infected with the A4557-5 virus were immune to challenge with virulent V154 virus. However, in comparison to SA-2 virus, this virus was a less effective immunizing agent when administered by the vent or drinking water methods.  相似文献   

13.
一株分离自外观正常鸡胚的禽呼肠孤病毒的鉴定   总被引:1,自引:0,他引:1  
从外观正常的鸡胚体内分离到一株病毒,其在鸡胚原代成纤维细胞上繁殖产生稳定的以合胞体为特征的CPE;病毒呈二十面体对称,无囊膜,有双层衣壳,直径为50~60nm;SDS-PAGE显示病毒基因组分为10个片段,呈3—3—1—3分布,与禽呼肠孤病毒(ARVS)FDO株基本相同。以上资料说明,鸡胚所带病毒为一株ARVS,其在CEF-2代上的TCID_(50)为10~(-6.16)/0.1ml。  相似文献   

14.
The growth of five strains of infectious bursal disease virus--three strains of serotype I (SAL, D-78, 2512), one of serotype II (OH), and one variant strain (Variant-A)--were compared in Vero and chicken embryo fibroblast (CEF) cell cultures in order to characterize the replication of different strains of IBDV in Vero cells. For all five virus strains, the latent period in Vero cells ranged from 12 to 18 hr, which was longer than the 4-to-6-hr latent period observed in CEF cultures for strains SAL, D-78, and OH. Virus strains SAL, D-78, and OH, which were examined in both Vero and CEF cultures, also had a more extensive maturation phase and higher yields of virus in Vero than in CEF cultures. Total titers of these viruses of 5.35 to 6.10 log10 TCID50/ml in CEFs occurred 24 to 30 hr postinoculation (PI), although the cytopathic effect (CPE) was not seen until 72 hr PI. By comparison, their total infectious virus titers of 6.85 to 8.35 log10 TCID50/ml in Vero cells occurred from 48 hr PI, coinciding with the appearance of CPE. The growth curve of Variant-A in Vero cells differed from the other viruses by showing steadily rising extracellular and cell-associated virus titers throughout the 72-hr observation period. Only very low titers of Variant-A were obtained in CEF cultures, and thus no growth curve in CEFs was performed.  相似文献   

15.
Three biological properties of canine distemper virus were examined to determine if any would consistently differentiate field from vaccine strains of the virus. The properties were the ability to (1) infect macrophages and epithelial cells, (2) produce distinct cytopathologic effect in alveolar and peritoneal macrophages and Vero cells, and (3) produce pocks on the chorioallantoic membrane of embryonated chicken eggs. Four vaccine strains and 5 field isolates were used in the study. The 5 field isolates were obtained directly from canine tissues. Of the 3 properties studied, only the comparison of the ability of the viruses to infect macrophages and epithelial cells was a consistent marker of virus origin. Virulent field isolates would only infect macrophage cultures, whereas the vaccine strains infected both types of cells. One avirulent field isolate from a case of old dog encephalitis reacted more like a vaccine strain by infecting both cell types.  相似文献   

16.
Nine serologic types of avian paramyxovirus (APMV) have been recognized. Newcastle disease virus (APMV-1) is the most extensively characterized virus, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two strains of APMV-2, Yucaipa and Bangor, in 9-day-old embryonated chicken eggs, 1-day-old specific-pathogen-free (SPF) chicks, and 4-wk-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was more than 168 hr for both strains, and their intracerebral pathogenicity index (ICPI) was zero, indicating that these viruses are nonpathogenic in chickens. When inoculated intracerebrally in 1-day-old chicks, neither strain caused disease or replicated detectably in the brain. This suggests that the zero ICPI value of APMV-2 reflects the inability of the virus to grow in neural cells. Groups of twelve 4-wk-old SPF chickens and turkeys were inoculated oculonasally with either strain, and three birds per group were euthanatized on days 2, 4, 6, and 14 postinoculation for analysis. There were no overt clinical signs of illnesses, although all birds seroconverted by day 6. The viruses were isolated predominantly from the respiratory and alimentary tracts. Immunohistochemistry studies also showed the presence of a large amount of viral antigens in epithelial linings of respiratory and alimentary tracts. There also was evidence of systemic spread even though the cleavage site of the viral fusion glycoprotein does not contain the canonical furin protease cleavage site.  相似文献   

17.
Seven of nine avian virus families tested (Birnaviridae, Coronaviridae, Herpesviridae, Paramyxoviridae, Poxviridae, Reoviridae, and Retroviridae) were found to replicate in a quail fibroblast cell line, designated QT35, resulting in a cytopathic effect (CPE) visible with the naked eye or by low-power microscopy. In comparison, only one (Paramyxoviridae) of seven mammalian virus families tested produced an observable CPE. Cytopathic changes induced by examined viruses were round cell, syncytial, and focus formation. Trypsin did not promote cytopathic changes by selected CPE-negative avian and mammalian viruses in QT35 cells. Several avian viruses (infectious bursal disease virus, Newcastle disease virus, Canary pox virus, and reovirus) formed plaques under agar. Avian reovirus and infectious bursal disease virus produced similar titers in chicken embryo fibroblast (CEF) and QT35 cell cultures. Chicken-egg-yolk neutralizing-antibody titers to IBDV were comparable in CEF and QT35 cell-culture systems.  相似文献   

18.
The Sersenk strain of goat pox virus first isolated in Iraq, grew on chorioallantoic membranes (CAM) of developing chicken embryos producing generalised large pocks, 2 to 3 mm in diameter, on the third day post-inoculation. The virus killed the inoculated embryos. Replication of the virus in primary cultures of lamb testis cells induced a cytopathic effect (CPE) and plaque formation characteristic of pox viruses. It was antigenically related to reference strains of goat pox and sheep pox viruses. The virus was sensitive to ether and chloroform, failed to agglutinate erythrocytes and was strictly pathogenic to goats.  相似文献   

19.
减蛋综合征病毒末端片段的克隆及细胞内DNA重组   总被引:6,自引:0,他引:6  
采用9~10日龄非免疫鸭胚增殖的减蛋综合征病毒(EDSV)AA-2毒株,经差速离心法浓缩纯化后,提取病毒基因组DNA。采用碱变性法除去病毒基因组共价结合的末端蛋白(TP)。用限制性内切酶HindⅢ水解纯化的EDSV基因组DNA。经低熔点琼脂糖凝胶电泳后,回收C、D、E片段。克隆到pUC19载体的HindⅢ和SmaⅠ双酶切位点及HindⅢ位点上,经蓝白斑筛选和单、双酶切鉴定,获得了pUHC、pUHD、pUHE重组质粒,其中pUHC含有末端片段。将EDSVSalⅠ水解产生并回收的大片段与pUHC在95℃水浴中变性,65℃复性后,用钙离子介导法,共转染50%~70%的单层鸭胚成纤维细胞,转染后36h开始产生细胞病变(CPE)。48h后将病变细胞反复冻融,经尿囊腔接种9~10日龄鸭胚,回收的尿囊液能凝集鸡红细胞,这种血凝性能被EDSV高免血清抑制,电镜下观察到腺病毒样颗粒。  相似文献   

20.
Eighty-nine avian reoviruses isolated from diseased and clinically normal chickens were classified serologically using antisera against five prototype strains. Eighty-three strains were classified into five serotypes; six strains were untypable. Most of the cytopathogenic strains that produced a clear cytopathic effect (CPE) in chicken embryo fibroblasts (CEFs) were highly pathogenic for chicken embryos (80% or more mortality via the allantoic sac) and for chicks (severe footpad swellings and tenosynovitis). These strains were classified into a single serotype represented by the TS-142 prototype strain. However, 10 strains that could not produce a clear CPE in CEFs showed very low pathogenicity for embryos and chicks, and these strains were serologically different from the TS-142 prototype strain. There was a strong relationship between pathogenicity and serotype. About 17% of the isolates were considered highly pathogenic.  相似文献   

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