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Molecular characterization and phylogenetic study of the fusion genes of Newcastle disease virus from the recent outbreaks in Ahvaz, Iran.

ABSTRACT: Newcastle disease (ND) is an acute and highly contagious disease affecting many domestic and wild species of birds. Its effects are most notable in domestic poultry due to their high susceptibility and the potential for severe impacts of an epizootic on the poultry industry. In this study, partial sequences of fusion genes of three Newcastle disease virus (NDV) isolates collected during 2013-14 outbreaks from the vaccinated commercial broiler chicken farms with high mortality around Ahvaz city (Southwest of Iran) were characterized. All three isolates showed the amino acid sequence 112RRQKRF117 at the C-terminus of the F2 protein and phenylalanine at the N-terminus of the F1 protein residue 117. These amino acid sequences were identical to a known virulent motif. The phylogenetic analysis revealed that the Iranian ND isolates in this study are closely related to the genotype VIId of class II NDV strains. Our results specified that there are velogenic NDV circulating in Ahvaz commercial broiler flocks and causing outbreaks in poultry industry.

SUBMITTER: Boroomand Z

PROVIDER: S-EPMC4758317 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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Project description:BackgroundAfrican swine fever (ASF) is a highly lethal and economically significant disease of domestic pigs in Eastern Africa particularly in Uganda where outbreaks regularly occur. Sequence analysis of variable genome regions have been extensively used for molecular epidemiological studies of African swine fever virus (ASFV) isolates. By combining p72, P54 and pB602L (CVR), a high level resolution approach is achieved for viral discrimination. The major aim of this study therefore, was to investigate the genetic relatedness of ASF outbreaks that occurred between 2010 and 2013 in Uganda to contribute to the clarification of the epidemiological situation over a four year period.MethodsTissue samples from infected domestic pigs associated with an ASF outbreak from 15 districts in Uganda were confirmed as being infected with ASFV using a p72 gene-based polymerase chain reaction amplification (PCR) assay recommended by OIE. The analysis was conducted by genotyping based on sequence data from three single copy ASFV genes. The E183L gene encoding the structural protein P54 and part of the gene encoding the p72 protein was used to delineate genotypes. Intra-genotypic resolution of viral relationships was achieved by analysis of tetramer amino acid repeats within the hypervariable CVR of the B602L gene.ResultsTwenty one (21) ASF outbreaks were confirmed by the p72 ASF diagnostic PCR, however; only 17 isolates were successfully aligned after sequencing. Our entire isolates cluster with previous ASF viruses in genotype IX isolated in Uganda and Kenya using p72 and P54 genes. Analysis of the CVR gene generated three sub-groups one with 23 tetrameric amino acid repeats (TRS) with an additional CAST sequence, the second with 22 TRS while one isolate Ug13. Kampala1 had 13 TRS.ConclusionWe identified two new CVR subgroups different from previous studies. This study constitutes the first detailed assessment of the molecular epidemiology of ASFV in domestic pigs in the different regions of Uganda.

Project description:BackgroundCarbapenem resistant Acinetobacter baumannii is an important nosocomial pathogen associated with a variety of infections.ObjectivesThe current study aimed to characterize the antimicrobial susceptibility, analyze the prevalence of oxacillinase and metallo-β-lactamase (MBL) genes and molecular typing of clinical isolates of A. baumannii.Materials and methodsA total of 124 non-repetitive isolates of A. baumannii were collected from various clinical specimens in two teaching hospitals in Ahvaz, south-west of Iran. Antimicrobial susceptibility test was carried out by disk diffusion method. The minimum inhibitory concentrations (MICs) of imipenem, meropenem, colistin and tigecycline were determined using E-test. To screen for MBL production, double disk synergy (DDs) test and MBL E-test were performed. The presence of bla OXA-23-like, bla OXA-24-like, bla OXA-51-like, bla OXA-58-like, bla VIM, bla IMP and bla SPM genes was assessed by polymerase chain reaction (PCR). To identify clonal relatedness, all isolates were subjected to repetitive sequence-based PCR (rep-PCR).ResultsBased on disk diffusion results, the highest rate of resistance was observed in rifampin (96.8%). Colistin and polymyxin-B were the most effective agents in vitro. According to the MIC results, the rate of resistance to imipenem, meropenem, colistin and tigecycline were 78.2%, 73.4%, 0.8% and 0, respectively. Metallo-β-lactamase production was positive in 42.3% and 79.4% of the isolates by DDs test and E-test, respectively. All isolates (100%) carried bla OXA-51-like gene. According to the results of multiplex PCR, bla OXA-23-like and bla OXA-24-like genes were detected in 85.6% and 6.2% of carbapenem resistant isolates, respectively. No bla OXA-58- like, bla VIM, bla IMP and bla SPM were detected. By rep-PCR, carbapenem resistant isolates were separated into six genotypes (A to F). Genotype A (30.9%) was the most prevalent (P value < 0.001). Genotypes B and C were found in 28.9% and 26.8% of the isolates, respectively.ConclusionsThe rate of carbapenem resistant A. baumannii isolates were high in this study. Since, bla OXA-58-like or MBL genes were not detected, it seems that resistance to carbapenems is related to bla OXA-23-like and bla OXA-24-like. Moreover, bla OXA-23-like was the most prevalent oxacillinase (OXA) gene. Most of the isolates belonged to one of the four dominant genotypes indicating clonal dissemination in the hospitals under study. In order to control the spread of carbapenem-resistant A. baumannii, infection- control strategies are needed.

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Molecular characterization and phylogenetic study of the fusion genes of Newcastle disease virus from the recent outbreaks in Ahvaz, Iran.

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