Original Scientific Article
Classical and molecular characterization of pigeon paramyxovirus type 1 (PPMV-1) isolated from backyard poultry – first report in Macedonia
Dodovski Aleksandar
*
,
Krstevski Kiril
,
Naletoski Ivancho
Abstract
Aim of this study was to characterize pigeon variant of Newcastle disease virus (NDV) isolated from backyard poultry using classical and molecular methods. In standard hemagglutination inhibition (HI) test both polyclonal NDV antiserum and monoclonal antibodies 161/617 specific for pigeon variants of NDV showed inhibition of heamagglutination of the isolated virus. Intracerebral pathogenicity index (ICPI) has shown that the isolate is mesogenic virus (ICPI = 0.81). One-step RT-qPCR for detection of M gene was performed indicating a presence of NDV and RT-qPCR for discrimination between lentogenic and velogenic strains based on F gene was also performed indicating a presence of virulent NDV. A portion of the F gene was amplified and sequenced for determination of virulence and phylogenetic characterization. The F protein cleavage site sequence of the isolate had multiple basic amino acids at residues 112–116 and a phenyl alanine at residue 117 (112RRQKR*F117) which is typical for velogenic strains. The nucleotide sequence of 374 bp was aligned to begin at nt 47 and finish at 420 immediately after the cleavage site and compared with other reference strains from the region and worldwide. In the phylogenetic tree, the isolate clustered into genotype VIb, typical for PPMV-1. This strain is phylogenetically very similar to other PPMV-1 isolated from pigeons in Macedonia. Poultry infected with PPMV-1 can spread the virus in the absence of clinical signs, thus PPMV-1’s are constant threat to domestic poultry. This is the first report of evidenced spillover of PPMV-1 into poultry in Macedonia.
Keywords: PPMV-1, backyard poultry, RT-qPCR, nucleotide sequencing
Acknowledgement
We would like to acknowledge: 1. Dr. Vladimir Savić from the Croatian Veterinary Institute, Poultry Centre, Zagreb, Croatia for the initial training in techniques performed in this study. 2. Dr. Dejan Vidanović from the Specialized Veterinary Institute Kraljevo, Serbia for provision of Serbian sequences. 3. Dr. Christian Grund from the National Reference Laboratory for Newcastle Disease, FLI-Riems, Germany for his generous help in performing the ICPI test. 4. Dr. Ruth Manvell from the Animal Health and Veterinary Laboratories Agency, Weybridge UK for supplying monoclonal antibodies used in this study.
Citation Information
Macedonian Veterinary Review. Volume 36, Issue 1, Pages 33-39, p-ISSN 1409-7621, e-ISSN 1857-7415, 2013
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