doi: 10.15389/agrobiology.2016.2.255rus

UDC 619:578.2(574)

 

MOLECULAR AND BIOLOGICAL PROPERTIES OF PATHOGENIC
NEWCASTLE DISEASE VIRUSES ISOLATED IN KAZAKHSTAN

M.B. Orynbaev, K.T. Sultankulova, A.A. Kerimbaev,
V.M. Strochkov, E.K. Shalgynbaev, Z.D. Omarova,
G.K. Musaeva, E.D. Burashev, Zh.K. Kydyrbaev, A.R. Sansyzbai

Research Institute for Biological Safety Problems SC MES RK,
p.g.t. Gvardeiskii, Kordaiskii Region, Zhambylskaya Province, 080409 Republic of Kazakhstan,
e-mail ribsp@biosafety.kz

Received September 22, 2015

 

Currently, Newcastle disease (ND) is highly contagious viral infection of birds, characterized by pneumonia, encephalitis, multiple pointed hemorrhages and defeat of internals is spread in various regions of the world. To the present, all isolated ND viruses (Paramyxoviridae, Paramyxovirus) are divided into two classes, representing the diverse and constantly developing group of viruses. Despite universal vaccination, the disease is difficult to control, and in connection with this the ND causal agent is listed among the most important pathogens. In recent years, studies on the genetic variability of the ND strains in Kazakhstan were not conducted, although many transcontinental migratory routes of wild birds, the main carriers of the pathogens, are crossed exactly here. The present study was conducted to examine the characteristics of the circulation, as well as the isolation and characterization of isolates of Newcastle disease virus that caused the disease of poultry in different regions of Kazakhstan in 2010, 2012 and 2013. Using 10-day-old developing chicken embryos (DCE), we studied virus isolates from dead hens at ND outbreaks in poultry farms and private yards in Almatinskaya, North Kazakhstan and Zhambylskaya provinces of Kazakhstan. The mean death time of embryos (MDT) and intracerebral pathogenicity index (ICPI) were estimated. Viral RNA was isolated and used for PCR. Amplified products were further detected, purified and sequenced. The obtained nucleotide sequences were analyzed using Sequencher v. 4.5 (Gene Codes Corporation, USA). A set of nucleotide sequences from an international database GenBank was used to construct the dendrogram and determine the genotype. Phylogenetic analysis of the sequences was performed using Mega 6.06 and the following parameters: Statistical Method — Neighbor-joining; Test of Phylogeny — Bootstrap method; No. of Bootstrap Replications — 500; Model/Method — Kimura 2-parameter model. Studies have shown that the ND virus causes outbreaks both among vaccinated and non-vaccinated poultry. The ND isolates belong to velogenic strains. All of them had a proteolytic cleavage site 110GGRRQKRF117 in the fusion protein, which is characteristic of the V-pathotype. The sequencing and phylogenetic analysis of the F-gene showed that the virus from dead birds of those vaccinated at the poultry farm in Almatinskaya Province belongs to VIId genotype, while the isolates from non-vaccinated birds of the private farms in Almatinskaya, Zhambylskaya and North Kazakhstan provinces belong to VIIb genotype. According to the obtained information, despite the geographical distance of outbreaks, the same ND virus genotypes are circulating in the territory of Northern Kazakhstan and in the southern regions of the country. Wide spread of the virus in Kazakhstan requires from veterinary services to develop effective control measures with regard to ND molecular epidemiology.   

Keywords: Newcastle disease, strain, pathogenicity index, PCR, sequencing, phylogenetic analysis.

 

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