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1.
Isolation of hemorrhagic enteritis virus (HEV) from spleens of infected turkeys in the MDTC-RP19 lymphoblastoid cell line was compared with detection of HEV antigen in the same spleens using the agar gel precipitation (AGP) test. A concordance of 80% was found between the two assays. Virus isolation had a sensitivity of 84% and a specificity of 88% compared with the AGP test. RP19 cells were also susceptible to infection with several other avian adenoviruses, but such infection was easily differentiated from that of HEV by a fluorescent-antibody (FA) test. Turkeys required 10(2) tissue-culture-infectious doses (TCID) to develop HE-specific lesions and 10(5) TCID to be killed. On the other hand, as little as 10 TCID of apathogenic HEV protected the poults against challenge with virulent HEV. The enzyme-linked immunosorbent assay (ELISA) for detection of HEV antibody was improved by using virus-infected RP19 cells as antigen. The ELISA appears to be more sensitive than the serum-neutralization test.  相似文献   

2.
Polypeptides of hemorrhagic enteritis virus (HEV) of turkeys and marble spleen disease virus (MSDV) of pheasants were analyzed by immune precipitation and immunoblot assays. A total of 11 polypeptides ranging in molecular weight from 14,000 to 97,000 were detected in lysates of HEV-infected turkey cells analyzed by immunoblot assay using a polyclonal antibody against HEV. Identical patterns were observed with preparations of MSDV. Five monoclonal antibodies (MAbs) against HEV were chosen based on their virus neutralization activity and used for identification of neutralizing epitopes of these two viruses. Three MAbs precipitated a single 97,000-molecular-weight hexon polypeptide in an immune precipitation assay.  相似文献   

3.
The MDTC-RP30 lymphoblastoid cell line established from Marek's disease (MD) tumors in turkeys consisted of a heterogeneous population of cells 10 to 25 micron in diameter. Large-cell fractions obtained from a bovine fetal serum gradient had a higher titer of cell-associated MD virus (MDV) than the small-cell fractions. Seven single-cell clones were established from MDTC-RP30 cell line: two consisted of large cells, and the other clones consisted of small cells. Infectious MDV was rescued from large-cell clones in chicken embryo fibroblast cultures but not from small-cell clones. All clones contained MDV DNA sequences when hybridized against cloned MDV DNA. All clones were positive for a Marek's-disease-tumor-associated surface antigen and surface immunoglobulins. All but two small-cell clones caused MD in susceptible chickens. The two large-cell-type clones were uniformly tetraploid, whereas one small-cell clone was diploid and the four others were a mixture of diploid and tetraploid, with an occasional triploid cell. Evidence of translocation involving the male (Z) chromosome and the chromosome #3 was seen in one clone. These results suggest that MDV transforms different subpopulations of lymphocytes.  相似文献   

4.
The effects of single and combined vaccination of turkeys against hemorrhagic enteritis virus (HEV) and Newcastle disease virus (NDV) were investigated. Dual vaccination of turkeys with NDV-B1 and HEVp30 or marble spleen disease virus (MSDV) enhanced white mottling of the spleens and the apoptosis rate in spleen cells (P < 0.05). In addition, simultaneously vaccinated turkeys had fewer HEV-infected spleen cells at 4 days postvaccination than turkeys given HEVp30 or MSDV alone. The anti-HEV antibody response was significantly reduced at 14 days postvaccination (P < 0.05), whereas the anti-NDV antibody response was enhanced (P < 0.05) in turkeys vaccinated with HEVp30 + NDV-B1. Further, the effect of dual vaccination on macrophage function was studied. Spleen cells from NDV-B1-vaccinated turkeys were primed to produce nitric oxide (NO) after stimulation in vitro with lipopolysaccharide. Spleen cells from HEVp30- or MSDV-vaccinated turkeys did not produce NO after in vitro stimulation. In dual-vaccinated turkeys, the priming effect of NDV-B1 was reduced in comparison with single-inoculated birds.  相似文献   

5.
Enzyme-linked immunosorbent assays (ELISAs) were developed to quantitate hemorrhagic enteritis virus (HEV) antibodies in turkey sera and HEV antigens in tissue extracts. These assays were more sensitive than the commonly used agar-gel precipitin tests in detecting antigen and antibody. The antibody-ELISA was used to monitor the presence and decline of passive antibodies in turkey poults and the seroconversion of turkeys infected with HEV. The antigen-ELISA was carried out using a monoclonal antibody; this test was used to quantitate HEV antigen in experimentally infected turkeys in a time-sequence experiment. Both ELISAs measured a strong antigenic relationship between an avirulent strain (HEV-A) and a virulent strain (HEV-V).  相似文献   

6.
Hemorrhagic diarrhea, gross hemorrhagic enteritis, and death caused by intravenous virus injection of hemorrhagic enteritis virus (HEV) were prevented in otherwise susceptible turkey poults by surgical splenectomy. The splenectomized poults produced anti-HEV antibodies, which indicated that splenectomy did not completely prevent replication of the virus. These results indicate that the spleen is necessary for the development of the intestinal lesions of this disease. The role of a toxic factor in this disease is discussed.  相似文献   

7.
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.  相似文献   

8.
The aim of the study was to determine the effect of a Polish low-virulence isolate of haemorrhagic enteritis adenovirus (HEV) on the immune system in turkeys and on the course of colibacillosis in birds infected under laboratory conditions. Turkeys were infected per os with HEV at the dose of 10(4.3)EID50/mL and with E. coli (APEC) (serotypes 078:K80:H9) at the dose of 4x10(9)CFU/mL by injection to the thoracic air sac. The birds infected with the HEV were infected with the APEC either simultaneously or after 5 days. Five days after HEV infection, the percentages of subpopulations of the CD3+CD4+ and CD3+CD8alpha+ T cells and the IgM+ B cells were determined in blood and spleens of the HEV-infected turkeys and in the control (uninfected) birds. The course of colibacillosis was more severe in turkeys infected with the APEC 5 days after infection with the HEV than in those infected with the HEV and APEC simultaneously and than in those infected only with APEC. Five turkeys out of the 18 infected with the APEC 5 days after infection with HEV, died. Their body weights were statistically significantly lower with higher FCR values 41 days after the infection in comparison to turkeys in the other groups. A considerable decrease in the percentage of the T and B cells subpopulations in the blood were found in turkeys infected with the HEV and while the percentage of CD3+CD4+ T cells subpopulation in the spleen increased significantly, the contribution of the CD3+CD8alpha+ T cells and IgM+ B cells subpopulations were decreased. These changes in the immune system of turkeys, occurring 5 days after infection with the HEV, made them more susceptible to infection with the APEC.  相似文献   

9.
Propagation of group II avian adenoviruses in turkey and chicken leukocytes   总被引:1,自引:0,他引:1  
An avirulent hemorrhagic enteritis virus isolate (HEV-A) as well as a virulent one (HEV-V), both belonging to the group II avian adenoviruses, were successfully propagated in turkey leukocyte cell cultures. HEV antigens were detected as early as 12 hr after infection of the cells, using HEV-specific monoclonal antibodies in a fluorescent antibody test, and virus particles were observed by electron microscopy in the nuclei of infected cells at 18 to 24 hr after infection. Electron microscopy revealed the presence of HEV in the nuclei of nonadherent cells, as well as in adherent cells. The nonadherent infected cells had the characteristics of immature mononuclear leukocytes, whereas the adherent cells had monocyte-macrophage characteristics. HEV produced in turkey leukocytes was mostly cell-associated, particularly with the nonadherent cells. HEV-A could be serially passed in turkey blood leukocyte cultures at least seven times. Various methods employed to culture virus indicated that cells grown in spinner cultures were superior to cells grown in stationary cultures. In contrast to the successful infection of HEV in turkey leukocytes, the infection of chicken leukocytes with either HEV or splenomegaly virus of chickens, or turkey leukocytes with splenomegaly virus, was poor.  相似文献   

10.
Commercial turkey eggs, free of antibodies to avian metapneumovirus subtype C (aMPV/C), were inoculated with aMPV/C at embryonation day (ED) 24. There was no detectable effect of virus inoculation on the hatchability of eggs. At 4 days post inoculation (DPI) (the day of hatch (ED 28)) and 9 DPI (5 days after hatch), virus replication was detected by quantitative RT-PCR in the turbinate, trachea and lung but not in the thymus or spleen. Mild histological lesions characterized by lymphoid cell infiltration were evident in the turbinate mucosa. Virus exposure inhibited the mitogenic response of splenocytes and thymocytes and upregulated gene expression of IFN-γ and IL-10 in the turbinate tissue. Turkeys hatching from virus-exposed eggs had aMPV/C-specific IgG in the serum and the lachrymal fluid. At 3 week of age, in ovo immunized turkeys were protected against a challenge with pathogenic aMPV/C.  相似文献   

11.
12.
A highly sensitive and specific double-antibody enzyme-linked immunosorbent assay (ELISA) is described for the detection of antigen and antibody of turkey hemorrhagic enteritis virus (HEV). The assay utilizes a virus-neutralizing monoclonal antibody (MAb) to capture the antigen and turkey antiserum against HEV as the second antibody. Microtiter plates were first coated with a dilution of 1:3000 of the MAb (300 ng immunoglobulin/well) and are used for detection of both antigen and antibody. For antibody detection, MAb-coated plates were treated with an appropriate dilution of a cell-culture-propagated HEV antigen and then reacted with the test turkey serum. For detection of HEV antigen, MAb-coated plates were treated with appropriate dilutions of test antigens and then reacted with purified anti-HEV turkey immunoglobulins. The assay for HEV antibody detection was more sensitive and specific than previously described single-antibody ELISAs. Using the double-antibody ELISA, it was found that the spleen of HEV-infected turkeys harbors very high levels of antigen. Traces of HEV antigen are present in some other organs. Infectivity assay for HEV is found to be about two orders of magnitude more sensitive than the ELISA for detection of virus.  相似文献   

13.
Localisation of swine hepatitis E virus in experimentally infected pigs   总被引:2,自引:0,他引:2  
The distribution of intravenously inoculated swine hepatitis E virus (HEV) was assessed by in situ hybridisation for a period of 50 days. Evidence of apparent clinical disease was found in only one pig in the HEV infected group. The only gross lesion observed was mildly enlarged mesenteric lymph nodes at 50 days post infection (dpi). Histopathologically, mild lymphoplasmacytic infiltration and focal hepatocellular necrotic lesions were found in HEV-infected pigs. Swine HEV nucleic acids were detected by RT-PCR in the faeces at 3 dpi in 100% of the 18 pigs infected with the virus. Thereafter, the number of positives declined.The most consistent and intense signal was found in the liver of infected animals using in situ hybridisation. The positive cells were hepatocytes, Kupffer cells, bile epithelial cells and interstitial lymphocytes. Swine HEV RNA was localised in the cytoplasm of the hepatocytes, with a slightly granular pattern of staining, but hybridisation signals were not observed in degenerative or vacuolated hepatocytes. HEV was much less frequently detected in extrahepatic tissues such as lymph nodes, tonsil, spleen and small and large intestine. It was concluded that swine HEV had replicated primarily in the hepatocytes and infection resulted in subclinical infection with minimal histopathological changes in the liver.  相似文献   

14.
Culture fluids from Marek's disease (MD) lymphoblastoid cell lines have suppressive activity against normal and mitogen-stimulated chicken spleen and bursal cells and also against the homologous cell lines. Suppressive activity was also present in supernatants from spleen cells infected in vitro with MD virus. The suppressor factor from MD cell lines was non-sedimentable, trypsin sensitive, heat resistant and partially dialysable. Preliminary studies suggest it has a molecular weight of 20,000 daltons. Studies were also conducted on the effect of the prostaglandin inhibitors indomethacin and aspirin on the production and action of the suppressor factor. At low concentrations they have a stimulatory effect on the cell lines suggesting that they inhibit the effects of suppressor factor; however only small amounts of prostaglandin E2 were present in supernatants. Evidence was obtained that the suppressor factor may act indirectly by stimulating the production of prostaglandin by spleen cell cultures. The role of a suppressor factor in the immunosuppression observed in MD is discussed.  相似文献   

15.
Avian pneumovirus (APV) is an immunosuppressive respiratory pathogen of turkeys. We examined the effect of APV infection on the vaccine efficacy of hemorrhagic enteritis virus (HEV) vaccines. APV was inoculated in 2-wk-old turkeys. Two or four days later, an attenuated HEV vaccine (HEVp30) or marble spleen disease virus (MSDV) vaccine were administered. Virulent HEV challenge was given 19 days after HEV vaccination. APV exposure compromised the ability of HEVp30 and MSDV to protect turkeys against virulent HEV. The protective index values were as follows: MSDV (100%) versus APV + MSDV (0%) (P < 0.05); HEVp30 (60%) versus APV + HEVp30 (30%) (P < 0.05) (Experiment I) and HEVp30 (56%) versus APV + HEVp30 (20%) (P < 0.05) (Experiment II). These data indicated that APV reduced the efficacy of HEV vaccines in turkeys.  相似文献   

16.
Twenty-two flocks of turkeys affected by enteric problems, with ages between 10 and 104 days and located in the Southern region of Brazil, were surveyed for turkey by PCR for turkey astrovirus type 2 (TAstV-2), turkey coronavirus (TCoV), hemorrhagic enteritis virus (HEV), rotavirus, reovirus, Salmonella spp., and Lawsonia intracellularis (Li) infections. Eleven profiles of pathogen combination were observed. The most frequently encountered pathogen combinations were TCoV-Li, followed by TCoV-TAstV-2-Li, TCoV-TastV-2. Only TCoV was detected as the sole pathogen in three flocks. Eight and 19 flocks of the 22 were positive for TAstV-2 and TCoV, respectively. Six were positive for Salmonella spp. and L. intracellularis was detected in 12 turkey flocks. Reovirus and HEV were not detected in this survey. These results throw new light on the multiple etiology of enteritis in turkeys. The implications of these findings and their correlation with the clinical signs are comprehensively discussed, illustrating the complexity of the enteric diseases.  相似文献   

17.
Twenty chicks, 12 turkey poults and 10 ducklings, all 5 weeks old were infected with 2 × 103.5 chick LD50 IBD virus to determine the course of the virus in the 3 poultry species. Uninfected control birds were kept separately. Two infected and 2 control birds/species were euthanized at time intervals between 3 and 168 hours post infection (pi). Sections of thymus, bursa of Fabricius, spleen, liver, kidney, proventriculus and ceacal tonsil were stained for the detection of IBD virus antigen using immunoperoxidase technique. IBD virus antigen positive cells stained reddish-brown and the amount of such cells in tissue sections were noted and scored. Stained cells were present in all organs examined for up to 168 hours pi in the 3 poultry species except ceacal tonsils of ducks at 72 and 120 hours pi. Antigen score was highest in chickens and least in ducks as reflected by average of total scores/sampling time of 12, 10.8 and 8 in chickens, turkeys and ducks respectively. Total antigen score/sampling time in infected chickens peaked twice; 24/48 and 144 hours pi, whereas such bi-phasic peaks were absent in turkeys and ducks. Range of total antigen score at different sampling times was 7–17.5 in chickens, 10–13 in turkeys and 7–10 in ducks indicative of marked viral replication in chickens. Presence of IBD viral antigen in organs of all 3 poultry species is indicative of infections. The innate ability of turkeys and ducks to prevent appreciable replication of IBD virus after infection requires further investigation.  相似文献   

18.
Nuclear inclusion bodies typical of the adenovirus group were widespread in in the spleen and other tissues of 8-week-old turkeys with severe respiratory disease and concomitant evidence of colisepticemia. Adenoviral virions were seen in affected nuclei of splenic tissue and in negatively stained preparations of ground spleen. In splenic tissue, inclusions were most prominent in reticular cells and macrophages in the periarterial lymphoid sheaths, the red pulp and the marginal zones of the periarteriolar reticular sheaths. Marked reticuloendothelial hyperplasia, lymphoid atrophy and granulocytic splenitis characterized the splenic changes. There were inclusions in the respiratory tract, intestinal tract, liver, kidney and pancreas. Inoculation of young turkeys, especially when immunosuppressed, resulted in evidence of infection and respiratory disease. The viruses produced cytopathic changes in primary turkey kidney cell cultures but did not affect embryonating chicken eggs. Concentrated viral suspensions induced precipitin lines in agar gel immunodiffusion tests with known antisera against known turkey adenoviruses but did not show an antigenic relationship to chicken adenoviruses.  相似文献   

19.
An avian influenza virus with surface antigens similar to those of fowl plague virus (Hav 1 Nav 2) was isolated in 1979 from 2 commercial turkey flocks in Central Texas. Two flocks in contact with these infected flocks developed clinical signs, gross lesions, and seroconversion but yielded no virus. This was the first recorded incidence of clinical avian influenza in Texas turkeys and only the second time that an agent with these surface antigens was isolated from turkeys in U.S.  相似文献   

20.
The effect of avian influenza virus (AIV) infection on the ability of turkeys to eliminate Pasteurella multocida from the respiratory tract was evaluated. Four-week-old turkeys were experimentally infected with an apathogenic AIV subtype (H5N2) by the oculonasal route and subsequently superinfected with P multocida (Urbach strain) by the intranasal route three days after infection with AIV. Quantitative clearance of P multocida from the trachea and lung was determined using a pour plate technique on samples collected at intervals after infection. Samples from turkeys which had been infected with AIV were found to yield more P multocida than those from turkeys which had not been infected with AIV. The numbers of P multocida increased in infected birds to a greater extent than in birds which had not been infected with the virus. The present study suggests that AIV infection may contribute to the increased numbers and a decreased clearance of P multocida in turkeys.  相似文献   

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