Monoclonal antibodies (MAbs) were produced against a vaccinal S1133 strain of avian reovirus. Cha... more Monoclonal antibodies (MAbs) were produced against a vaccinal S1133 strain of avian reovirus. Characterization of six MAbs in Western blotting, radioimmunopre‐cipitation and gold immunoelectron microscopy revealed that the MAbs were specific to the outer capsid proteins, μ2/μ2c, σ2 and σ3. Two of three MAbs, directed against σ2 protein, neutralized the virus infectivity in a broadly specific manner, whereas the third
Three unrecognized field isolates of Infectious Bronchitis Virus (IBV) were recovered from commer... more Three unrecognized field isolates of Infectious Bronchitis Virus (IBV) were recovered from commercial broiler chickens vaccinated with live Mass viral strain (H120). These isolates were identified by immunofluorescence using monoclonal antibodies produced against reference serotypes: Mass, Conn, and Ark. RT-PCRs were performed on viral RNAs to amplify S1 gene using a specific set of primers S1OLIGO3' and S1OLIGO5'. Restriction polymorphism (RFLP) of PCR products was determined by the use of HaeIII restriction enzyme. As expected, patterns of PCR products were different from common pattern of strains assigned to Mass serotype M41, Beaudette, H120, and Florida. Molecular analysis showed a nucleotide insertion in hypervariable region one (HVR-1) of S1 gene of only Quebec isolates (Qu16, Qu_mv and Q_37zm). However, New Brunswick IBV isolate (NB_cp) did not display these insertions. Major amino acid changes involved insertion of two stretches (aa118-119: Arg-Ser and aa141-145: Sy...
The use of attenuated vaccines or the occurrence of low virulent T-lymphotropic or B-lymphotropic... more The use of attenuated vaccines or the occurrence of low virulent T-lymphotropic or B-lymphotropic viruses in flocks may alter the immune responses of young chicks in spite of the absence of clinical signs. Infections with a low virulent T-lymphotropic chicken infectious anaemia virus (lvCIAV) followed by infection with an intermediate B-lymphotropic infectious bursal disease virus (iIBDV) were conducted in specific pathogen free chicks. Thirty-six 1-day-old chicks were infected with the lvCIAV strain (CAV-VAC®) and a similar number of chicks were inoculated with phosphate-buffered saline. At 14 days after lvCIAV infection, one group of 18 lvCIAV-infected chicks and one group of 18 uninfected chicks were infected with an iIBDV strain. At 4, 7 and 14 days post infection with iIBDV, six chicks from each group were euthanized and lymphoid organs were collected. Detection of lvCIAV and iIBDV genomes was conducted by polymerase chain reaction and reverse transcriptase-polymerase chain reaction, respectively. Double-labelled lymphoid subsets from the thymus, spleen and bursa were studied by cytofluorometric analysis. The results reveal that previous infection with lvCIAV increases the occurrence of the lvCIAV and iIBDV genome in thymus and/or bursa without the occurrence of clinical signs in dually lvCIAV/iIBDV-infected chicks. However, the decreases of B cells in spleen and bursa and increases of T-cell subsets in bursa observed in chicks infected with iIBDV did not occur in chicks previously infected with lvCIAV. Taken together, these results suggest that previous infection of young chicks with lvCIAV decreases lymphoid disorders induced by iIBDV while subsequent iIBDV infection increases the lvCIAV genome in lymphoid organs.
Four chicken lymphoblastoid cell lines were inoculated with avian reovirus strain S1133 and two l... more Four chicken lymphoblastoid cell lines were inoculated with avian reovirus strain S1133 and two local isolates, 965 and 615. Of the inoculated cell lines, TLT, a B-cell line, was productively infected with the three viruses as demonstrated by immunofluorescence assay (IFA) and radioimmunoprecipitation assay. A comparative growth curve analysis of the three avian reoviruses was done at 37 degrees and 41 degrees C. Isolate 965 replicated to a higher titre at both temperatures while the replication of S1133 and 615 was found to be inhibited at 41 degrees C. IFA revealed that among the transformed T lymphoblastoid cells used in this study, only MDCC-RP1 was permissive to virus infection with isolate 965, and at 41 degrees C, but not 37 degrees C.
Mycoplasma gallisepticum (MG) is one of the aetiologic agents of chronic respiratory disease in c... more Mycoplasma gallisepticum (MG) is one of the aetiologic agents of chronic respiratory disease in chickens and infectious sinusitis in turkeys. We investigated humoral and cellular immune mechanisms following experimental infection with four different strains of MG. Peripheral blood leukocytes (PBL) obtained from chickens were examined for proliferation using antigen preparations of whole cell MG as stimuli in vitro. A consistent lymphoproliferative response was observed against the homologous whole cell antigens in the group of chickens infected with strain PG31. Significant lymphoproliferation was detected as early as 1 week post-infection. We further characterized antigen-specific proliferation by measuring the production of interferon and nitric oxide by the PBL of infected chickens. Consistent with lymphoproliferation, we also detected the presence of interferon and nitric oxide in vitro in antigen-stimulated cultures. These results indicate a possible role of cell-mediated immune responses in the development of immunity following MG infection in chickens.
Reoviruses were isolated from intestinal contents of broiler chickens from nine flocks in Quebec ... more Reoviruses were isolated from intestinal contents of broiler chickens from nine flocks in Quebec with malabsorption syndrome. Serum neutralization test demonstrated the existence of antigenic differences between the isolates and the reference vaccine strain. The isolated reoviruses were inoculated orally and into the foot pad in one-day-old chicks, resulting in a transient, but significant depression in body weight gains. Chickens infected with isolate 615, showed in addition to growth problems, clinical signs and tissue lesions similar to those observed in field cases. When isolate 615 was inoculated into SPF chicks at one day of age, intestinal absorption of D-xylose in infected chicks at 7 days post-infection was significantly lower (P <0.05) than for corresponding controls. This study suggests the implication of some reovirus isolates, such as 615 which was serologically distinct from the vaccine strain S1133, as infectious agents associated with pathological conditions other than viral arthritis.
Growth rate in broiler birds has increased substantially in the last decade due to improvement in... more Growth rate in broiler birds has increased substantially in the last decade due to improvement in genetics, feed formulation, cleaner environment, and vaccine formulations. As a result, it has become necessary to review and revise prediction method for vaccination in chicks. This study was undertaken to determine the possible use of the rate of weight gain rather than age in predicting vaccination time. Two groups of 1-day-old broilers originating from old and young breeders, respectively, and with different levels of maternal antibodies against infectious bursal disease virus (IBDV) were used in this study. The chicks were divided into four groups and subjected to two feed regiments: groups A1 and B1 were fed broiler feed for normal growth rate, and groups A2 and B2 were fed breeder feed for slower growth rate. At 1, 4, 8, 12, 16, 22, 29, and 36 days of age, 22 chicks in each group were weighed, and blood samples were collected. Serum samples were tested for antibodies against IBDV by enzyme-linked immunosorbent assay (ELISA) and virus neutralization test. Maternal antibody decline curves for each group were plotted according to chick age and chick weight. Fast-growing birds in groups A1 and B1 showed a faster rate of antibody decline, whereas slow-growing birds in groups A2 and B2 had a slower rate of antibody decline. Based on the effect of weight gain on maternal antibody decline, a new way of predicting vaccination time for IBDV based on measuring maternal antibody titers at 4 days of age was proposed and tested. The predicted antibody decline was shown to correspond to the real ELISA titers measured in our experiments (R = 0.9889), whereas a lower correlation (R = 0.8355) was detected between real ELISA titers and the titers predicted by the current method using age-based Deventer formula.
Monoclonal antibodies (MAbs) were produced against a vaccinal S1133 strain of avian reovirus. Cha... more Monoclonal antibodies (MAbs) were produced against a vaccinal S1133 strain of avian reovirus. Characterization of six MAbs in Western blotting, radioimmunopre‐cipitation and gold immunoelectron microscopy revealed that the MAbs were specific to the outer capsid proteins, μ2/μ2c, σ2 and σ3. Two of three MAbs, directed against σ2 protein, neutralized the virus infectivity in a broadly specific manner, whereas the third
Three unrecognized field isolates of Infectious Bronchitis Virus (IBV) were recovered from commer... more Three unrecognized field isolates of Infectious Bronchitis Virus (IBV) were recovered from commercial broiler chickens vaccinated with live Mass viral strain (H120). These isolates were identified by immunofluorescence using monoclonal antibodies produced against reference serotypes: Mass, Conn, and Ark. RT-PCRs were performed on viral RNAs to amplify S1 gene using a specific set of primers S1OLIGO3' and S1OLIGO5'. Restriction polymorphism (RFLP) of PCR products was determined by the use of HaeIII restriction enzyme. As expected, patterns of PCR products were different from common pattern of strains assigned to Mass serotype M41, Beaudette, H120, and Florida. Molecular analysis showed a nucleotide insertion in hypervariable region one (HVR-1) of S1 gene of only Quebec isolates (Qu16, Qu_mv and Q_37zm). However, New Brunswick IBV isolate (NB_cp) did not display these insertions. Major amino acid changes involved insertion of two stretches (aa118-119: Arg-Ser and aa141-145: Sy...
The use of attenuated vaccines or the occurrence of low virulent T-lymphotropic or B-lymphotropic... more The use of attenuated vaccines or the occurrence of low virulent T-lymphotropic or B-lymphotropic viruses in flocks may alter the immune responses of young chicks in spite of the absence of clinical signs. Infections with a low virulent T-lymphotropic chicken infectious anaemia virus (lvCIAV) followed by infection with an intermediate B-lymphotropic infectious bursal disease virus (iIBDV) were conducted in specific pathogen free chicks. Thirty-six 1-day-old chicks were infected with the lvCIAV strain (CAV-VAC®) and a similar number of chicks were inoculated with phosphate-buffered saline. At 14 days after lvCIAV infection, one group of 18 lvCIAV-infected chicks and one group of 18 uninfected chicks were infected with an iIBDV strain. At 4, 7 and 14 days post infection with iIBDV, six chicks from each group were euthanized and lymphoid organs were collected. Detection of lvCIAV and iIBDV genomes was conducted by polymerase chain reaction and reverse transcriptase-polymerase chain reaction, respectively. Double-labelled lymphoid subsets from the thymus, spleen and bursa were studied by cytofluorometric analysis. The results reveal that previous infection with lvCIAV increases the occurrence of the lvCIAV and iIBDV genome in thymus and/or bursa without the occurrence of clinical signs in dually lvCIAV/iIBDV-infected chicks. However, the decreases of B cells in spleen and bursa and increases of T-cell subsets in bursa observed in chicks infected with iIBDV did not occur in chicks previously infected with lvCIAV. Taken together, these results suggest that previous infection of young chicks with lvCIAV decreases lymphoid disorders induced by iIBDV while subsequent iIBDV infection increases the lvCIAV genome in lymphoid organs.
Four chicken lymphoblastoid cell lines were inoculated with avian reovirus strain S1133 and two l... more Four chicken lymphoblastoid cell lines were inoculated with avian reovirus strain S1133 and two local isolates, 965 and 615. Of the inoculated cell lines, TLT, a B-cell line, was productively infected with the three viruses as demonstrated by immunofluorescence assay (IFA) and radioimmunoprecipitation assay. A comparative growth curve analysis of the three avian reoviruses was done at 37 degrees and 41 degrees C. Isolate 965 replicated to a higher titre at both temperatures while the replication of S1133 and 615 was found to be inhibited at 41 degrees C. IFA revealed that among the transformed T lymphoblastoid cells used in this study, only MDCC-RP1 was permissive to virus infection with isolate 965, and at 41 degrees C, but not 37 degrees C.
Mycoplasma gallisepticum (MG) is one of the aetiologic agents of chronic respiratory disease in c... more Mycoplasma gallisepticum (MG) is one of the aetiologic agents of chronic respiratory disease in chickens and infectious sinusitis in turkeys. We investigated humoral and cellular immune mechanisms following experimental infection with four different strains of MG. Peripheral blood leukocytes (PBL) obtained from chickens were examined for proliferation using antigen preparations of whole cell MG as stimuli in vitro. A consistent lymphoproliferative response was observed against the homologous whole cell antigens in the group of chickens infected with strain PG31. Significant lymphoproliferation was detected as early as 1 week post-infection. We further characterized antigen-specific proliferation by measuring the production of interferon and nitric oxide by the PBL of infected chickens. Consistent with lymphoproliferation, we also detected the presence of interferon and nitric oxide in vitro in antigen-stimulated cultures. These results indicate a possible role of cell-mediated immune responses in the development of immunity following MG infection in chickens.
Reoviruses were isolated from intestinal contents of broiler chickens from nine flocks in Quebec ... more Reoviruses were isolated from intestinal contents of broiler chickens from nine flocks in Quebec with malabsorption syndrome. Serum neutralization test demonstrated the existence of antigenic differences between the isolates and the reference vaccine strain. The isolated reoviruses were inoculated orally and into the foot pad in one-day-old chicks, resulting in a transient, but significant depression in body weight gains. Chickens infected with isolate 615, showed in addition to growth problems, clinical signs and tissue lesions similar to those observed in field cases. When isolate 615 was inoculated into SPF chicks at one day of age, intestinal absorption of D-xylose in infected chicks at 7 days post-infection was significantly lower (P <0.05) than for corresponding controls. This study suggests the implication of some reovirus isolates, such as 615 which was serologically distinct from the vaccine strain S1133, as infectious agents associated with pathological conditions other than viral arthritis.
Growth rate in broiler birds has increased substantially in the last decade due to improvement in... more Growth rate in broiler birds has increased substantially in the last decade due to improvement in genetics, feed formulation, cleaner environment, and vaccine formulations. As a result, it has become necessary to review and revise prediction method for vaccination in chicks. This study was undertaken to determine the possible use of the rate of weight gain rather than age in predicting vaccination time. Two groups of 1-day-old broilers originating from old and young breeders, respectively, and with different levels of maternal antibodies against infectious bursal disease virus (IBDV) were used in this study. The chicks were divided into four groups and subjected to two feed regiments: groups A1 and B1 were fed broiler feed for normal growth rate, and groups A2 and B2 were fed breeder feed for slower growth rate. At 1, 4, 8, 12, 16, 22, 29, and 36 days of age, 22 chicks in each group were weighed, and blood samples were collected. Serum samples were tested for antibodies against IBDV by enzyme-linked immunosorbent assay (ELISA) and virus neutralization test. Maternal antibody decline curves for each group were plotted according to chick age and chick weight. Fast-growing birds in groups A1 and B1 showed a faster rate of antibody decline, whereas slow-growing birds in groups A2 and B2 had a slower rate of antibody decline. Based on the effect of weight gain on maternal antibody decline, a new way of predicting vaccination time for IBDV based on measuring maternal antibody titers at 4 days of age was proposed and tested. The predicted antibody decline was shown to correspond to the real ELISA titers measured in our experiments (R = 0.9889), whereas a lower correlation (R = 0.8355) was detected between real ELISA titers and the titers predicted by the current method using age-based Deventer formula.
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