Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons

Document Type : Original Article

Authors

1 Avian and Rabbit Medicine Department, Faculty of Veterinary Medicine, Zagazig University, 45511, Egypt

2 Veterinary Hospital, Faculty of Veterinary Medicine, Zagazig University, 45511, Egypt

3 Avian and Rabbit Diseases, Faculty of Veterinary Medicine, Zagazig University, Sharkia, Egypt

Abstract

Fifty-one pigeon houses from both commercial lofts and backyard of different ages and breeds were investigated for incrimination of pigeon paramyxovirus-1 (PPMV-1) and/ or Newcastle Disease virus (NDV) during 2013-2015. The results of clinical examination revealed nervous signs (wing paralysis, head tremors, torticollis, opisthotonos and leg paralysis), greenish diarrhea, and respiratory signs, with variable mortalities. The gross lesions included congestion and hemorrhage in brain; nephrosis and/ or nephritis in kidneys, ulcer in intestine and mucoid enteritis. Virus isolation via chicken embryo inoculation revealed embryo congestion and hemagglutination (HA) activity of harvested allantoic fluids were detected 41 samples (80.39%) with HA titers ranging from 26- 210. The hemagglutination inhibition (HI) results of the same isolates against PPMV-1 and NDV hyper immune sera which prepared in rabbits (1:5dilution) varied from 23 – 210   in 38 virus isolates. The allantoic fluids of 41 haemaglutinating virus isolates were subjected to Reverse transcriptase polymeraze chain reaction (RT-PCR), Thirty-eight isolates evidenced successful products for partial amplification of fusion protein gene of NDV and PPMV-1 at 356 bp using specific primers.  Restriction fragment length polymorphism (RFLP) using   a set of three enzymes; HhaI, MspI and MboI. revealed that 9 strains (mesogenic / lentogenic PPMV-1), 27(mesogenic / lentogenic NDV), one velogenic strain for PPMV-1 and the other was NDV. MDT value in ECE revealed 72 ± 0.0 and 62.4±5.89 indicating that both isolates could be mesogenic. It could be concluded that both NDV and PPMV-1 are co- circulating among pigeons and causing economical losses. Rt-PCR followed by RFLP can be useful rapid tools for detection and identification of both viruses. Beside, in the view of the failure of adopted   prophylactic vaccination, both LaSota and Inactivated PPMV-1 vaccines must be included to overcome the impact of aforementioned disease problems in pigeons

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