Isolation and characterization of avian reovirus associated with arthritis in broiler breeders
Latifa Bekheet Mohamed Bekheet;
Abstract
From broiler breeder flock in Giza governorate, demonstrating symptoms of arthritis, lameness and swelling of the hock joints, samples of tibiotarsal tendons, tendon sheaths and fluids around ruptured tendons were collected.
Homogenate of the samples was prepared then the filtrate was used for isolation of field avian reovirus by inoculation on the chorioallantoic membrane of specific pathogen-free embryonated chicken eggs (SPF-ECE) at 11-13 days old then propagation of the isolated virus was carried out for 6 passages in (SPF-ECE). CAM of the inoculated eggs showed pock lesions.
Filtrate prepared from the harvested CAMs was examined by haemagglutination test and AGPT for detection of contamination with other viruses than avian reovirus using specific antisera to such viruses.
Virus titration test was performed in SPF-ECE using filtrate of the harvested CAM from the sixth passage that showing pock lesions and the titer of the isolated virus was 106.1 EID50/ml.
For the antigenic characterization of the isolated virus, AGPT and FAT were conducted using antibodies specific to ARV and both tests gave positive results.
The VNT was conducted and the results revealed the antigenic relatedness between the isolated virus and ARV.
Transmission electron microscopic examination (negative staining) was applied on filtrate of pock lesion preparations, the results showed the virus particles in aggregates with apparent damage to the outer capsid of the virus.
Histopathological examination of pock lesions on the harvested CAM was carried out, the pathognomonic intracytoplasmic inclusion bodies were appeared.
RT-PCR was employed for molecular characterization of the isolated virus using primers specific to the conserved fragment of sigma C gene to amplify fragment with molecular size of 532 bp. Electrophoresis of the amplified products in agarose gel revealed the expected band with correct molecular size confirming the molecular characterization of the isolated virus.
The isolated virus designated as ARV/EGYPT/GIZA 2011. Further studies including sequencing of the amplified gene and the phylogenetic analysis will be conducted.
Homogenate of the samples was prepared then the filtrate was used for isolation of field avian reovirus by inoculation on the chorioallantoic membrane of specific pathogen-free embryonated chicken eggs (SPF-ECE) at 11-13 days old then propagation of the isolated virus was carried out for 6 passages in (SPF-ECE). CAM of the inoculated eggs showed pock lesions.
Filtrate prepared from the harvested CAMs was examined by haemagglutination test and AGPT for detection of contamination with other viruses than avian reovirus using specific antisera to such viruses.
Virus titration test was performed in SPF-ECE using filtrate of the harvested CAM from the sixth passage that showing pock lesions and the titer of the isolated virus was 106.1 EID50/ml.
For the antigenic characterization of the isolated virus, AGPT and FAT were conducted using antibodies specific to ARV and both tests gave positive results.
The VNT was conducted and the results revealed the antigenic relatedness between the isolated virus and ARV.
Transmission electron microscopic examination (negative staining) was applied on filtrate of pock lesion preparations, the results showed the virus particles in aggregates with apparent damage to the outer capsid of the virus.
Histopathological examination of pock lesions on the harvested CAM was carried out, the pathognomonic intracytoplasmic inclusion bodies were appeared.
RT-PCR was employed for molecular characterization of the isolated virus using primers specific to the conserved fragment of sigma C gene to amplify fragment with molecular size of 532 bp. Electrophoresis of the amplified products in agarose gel revealed the expected band with correct molecular size confirming the molecular characterization of the isolated virus.
The isolated virus designated as ARV/EGYPT/GIZA 2011. Further studies including sequencing of the amplified gene and the phylogenetic analysis will be conducted.
Other data
| Title | Isolation and characterization of avian reovirus associated with arthritis in broiler breeders | Other Titles | عزل و توصيف لفيروس الريو المصاحب لالتهاب المفاصل فى أمهات التسمين | Authors | Latifa Bekheet Mohamed Bekheet | Issue Date | 2016 |
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