Isolation and identification of Newcastle disease viruses from field outbreaks in chickens and pigeons

Abstract

Eleven dead or sick birds submitted from farms in the year 2010 with a history of sudden death with respiratory and/or diarrhoeal signs were used for isolation and identification of Newcastle disease virus (NDV). All samples were subjected to routine necropsy. Pooled respiratory tissues were inoculated in embryonated chicken eggs and chicken embryo fibroblast (CEF) cell culture. The growth of NDV was confirmed by embryo mortality, cytopathic effects (CPE) in cell culture, haemagglutination (HA) and haemagglutination inhibition (HI) test. The presence of NDV was confirmed by reverse transcriptionpolymerase chain reaction (RT-PCR). At necropsy seven cases were tentatively diagnosed as Newcastle disease (ND). Out of seven ND-suspected samples, four yielded virus in both embryos and cell culture, while one was positive only in embryos, one only in cell culture and one sample was negative in both embryos and cell culture. RT-PCR successfully amplified a 766 bp fragment covering parts of Matrix and Fusion protein genes of NDV from the samples that were positive either in embryos or in cell culture. It is suggested that RT-PCR could be a rapid and sensitive tool for the detection of NDV.

Immunomodulatory activity of curcumin-entrapped poly d,l -lactic- co -glycolic acid nanoparticles in mice

“…The anti-sera obtained from the blood were titrated against the erythrocyte antigen (1% SRBCs) in a 96-well microtitre plate according to Mazumder et al 26 Briefly, twofold serial dilution of anti-serum (50 μL) was made in 25 μL of 0.9% pyrogen-free normal saline. Thereafter, 25 μL of 1% SRBCs was added to each well, and microtitre plates were mixed gently by handshaking and incubated at room temperature (25°C) for two hours.…”

The positive expression of genotype VII Newcastle disease virus (Malaysian isolate) in Japanese quails (Coturnix coturnix japonica)

“…There was no attempt to isolate the virus by egg inoculation as the negative detection of virus in cloacal swabs due to the low replication of NDV in quails as mentioned by Mohamed et al , [13]. RT-qPCR was done in this study as it offers a reliable tool for quick detection of NDV, and this was agreed by Mazumder et al , 2012, which has performed the RT-PCR in his study for NDV detection although the gold standard is to perform virus isolation and serological identification followed by in vivo pathogenicity testing [8]. …”

Pathotypic and Genotypic Characterization of Two Bangladeshi Isolates of Newcastle Disease Virus of Chicken and Pigeon Origin

“…All the chickens suffered from respiratory distress with or without diarrhoea and the pigeon were highly depressed and anorexic before death. At necropsy, haemorrhagic lesions in the proventriculus with or without button like ulcers in the intestinal mucosa and congestion in lungs were found (Mazumder et al., ). Isolation of virus was attempted in chicken embryos and primary chicken embryo fibroblast (CEF) cell culture.…”

“…Six samples (five from chickens and one from a pigeon) resulted in embryo mortality and produced cytopathic effects (CPE) in CEF cell culture. All the six isolates were identified as NDV by haemagglutination and haemagglutination inhibition tests (Mazumder et al., ). Of these six field isolates of NDV, two isolates, one from a 21‐day‐old broiler chicken (APMV1/chicken/Bangladesh/BD‐C162/2010) and one from a 6‐month‐old domestic pigeon (APMV1/pigeon/Bangladesh/BD‐P01/2010), were selected for pathogenic and genetic characterization.…”