Molecular analysis of South African ovine herpesvirus 2 strains based on selected glycoprotein and tegument genes

Ovine herpesvirus 2 (OvHV-2), is the causative agent of sheep-associated malignant catarrhal fever (SA-MCF), a generally fatal disease of cattle and other captive wild ruminants. Information on the OvHV-2 strains circulating in South Africa (SA) and other African countries with regard to genetic structure and diversity, and pattern of distribution is not available. This study aimed to characterize the OvHV-2 strains circulating in SA using selected genes encoding glycoproteins and tegument proteins. To establish the genetic diversity of OvHV-2 strains, four genes, Ov 7, Ov 8 ex2, ORF 27 and ORF 73 were selected for analysis by PCR and DNA sequencing. Nucleotide and amino acid multiple sequence analyses revealed two genotypes for ORF 27 and ORF 73, and three genotypes for Ov 7 and Ov 8 ex2, randomly distributed throughout the regions. Ov 7 and ORF 27 nucleotide sequence analysis revealed variations that distinguished SA genotypes from those of reference OvHV-2 strains. Epitope mapping analysis showed that mutations identified from the investigated genes are not likely to affect the functions of the gene products, particularly those responsible for antibody binding activities associated with B-cell epitopes. Knowledge of the extent of genetic diversity existing among OvHV-2 strains has provided an understanding on the distribution patterns of OvHV-2 strains or genotypes across the regions of South Africa. This can facilitate the management of SA-MCF in SA, in terms of introduction of control measures or safe practices to monitor and control OvHV-2 infection. The products encoded by the Ov 7, Ov 8 ex2 and ORF 27 genes are recommended for evaluation of their coded proteins as possible antigens in the development of an OvHV-2 specific serodiagnostic assay.S1 Table. Average sequence identities determined for the Ov 7 nucleotide and amino sequences obtained between South African Ov 7 sequences compared to reference sequences.S2 Table. Average sequence identities determined for the Ov 8 ex2 nucleotide (Figure A) and derived amino acid (Figure B) sequences obtained between South African OvHV-2 strains compared to reference strains.S3 Table. Average sequence identities determined for the ORF 27 nucleotide and derived amino acid sequences obtained between South African OvHV-2 strains compared to reference strains.S4 Table. Average sequence identities for the ORF 73 nucleotide and derived amino acid sequences obtained between South African OvHV-2 strains compared to reference strains.This work was supported by Department of Science and Technology (DST) for research funding; Meat Industry Trust (MIT) for payment of the University tuitions.http://www.plosone.orgam2017Veterinary Tropical Disease

Molecular analysis of South African ovine herpesvirus 2 strains based on selected glycoprotein and tegument genes

Ovine herpesvirus 2 (OvHV-2), is the causative agent of sheep-associated malignant catarrhal fever (SA-MCF), a generally fatal disease of cattle and other captive wild ruminants. Information on the OvHV-2 strains circulating in South Africa (SA) and other African countries with regard to genetic structure and diversity, and pattern of distribution is not available. This study aimed to characterize the OvHV-2 strains circulating in SA using selected genes encoding glycoproteins and tegument proteins. To establish the genetic diversity of OvHV-2 strains, four genes, Ov 7, Ov 8 ex2, ORF 27 and ORF 73 were selected for analysis by PCR and DNA sequencing. Nucleotide and amino acid multiple sequence analyses revealed two genotypes for ORF 27 and ORF 73, and three genotypes for Ov 7 and Ov 8 ex2, randomly distributed throughout the regions. Ov 7 and ORF 27 nucleotide sequence analysis revealed variations that distinguished SA genotypes from those of reference OvHV-2 strains. Epitope mapping analysis showed that mutations identified from the investigated genes are not likely to affect the functions of the gene products, particularly those responsible for antibody binding activities associated with B-cell epitopes. Knowledge of the extent of genetic diversity existing among OvHV-2 strains has provided an understanding on the distribution patterns of OvHV-2 strains or genotypes across the regions of South Africa. This can facilitate the management of SA-MCF in SA, in terms of introduction of control measures or safe practices to monitor and control OvHV-2 infection. The products encoded by the Ov 7, Ov 8 ex2 and ORF 27 genes are recommended for evaluation of their coded proteins as possible antigens in the development of an OvHV-2 specific serodiagnostic assay.S1 Table. Average sequence identities determined for the Ov 7 nucleotide and amino sequences obtained between South African Ov 7 sequences compared to reference sequences.S2 Table. Average sequence identities determined for the Ov 8 ex2 nucleotide (Figure A) and derived amino acid (Figure B) sequences obtained between South African OvHV-2 strains compared to reference strains.S3 Table. Average sequence identities determined for the ORF 27 nucleotide and derived amino acid sequences obtained between South African OvHV-2 strains compared to reference strains.S4 Table. Average sequence identities for the ORF 73 nucleotide and derived amino acid sequences obtained between South African OvHV-2 strains compared to reference strains.This work was supported by Department of Science and Technology (DST) for research funding; Meat Industry Trust (MIT) for payment of the University tuitions.http://www.plosone.orgam2017Veterinary Tropical Disease

Analysis of the overhead throwing motion in softball and baseball

<p>Ref indicates the nucleotide sequences of the ORF 73 gene of the OvHV-2 reference genomes obtained from the GenBank. Variable regions are highlighted in rectangles (▯), and groups of genotypes (G) are illustrated on the right side of the align sequences.</p

Summary of genotypes of different genes investigated for characterization of South African OvHV-2 strains.

<p>Summary of genotypes of different genes investigated for characterization of South African OvHV-2 strains.</p

The phylogenetic trees of the O RF 73 nucleotide (A & B) and amino acid (C & D) sequences of OvHV-2 strains and published closely related reference sequences obtained from GenBank constructed using Neighbor-Joining (A & C) and Maximum Likelihood (B & D) methods.

<p>The unrooted condensed trees were constructed using Neighbor-Joining (A & C) and Maximum Likelihood (B & D) methods, showing branching patterns of each clade. Groups 1 and 2 are illustrated and abbreviated as clade 1 (C1) and 2 (C2), respectively, with supporting bootstrap values at each clade nodes.</p

Multiple sequence alignment of nucleotide sequences of the Ov 7 gene from SA OvHV-2 strains and reference sequences derived from OvHV-2 genome sequences (Accession numbers: NC-007646.1, AY839756.1, DQ198083.1), showing three genotypes of Ov 7 gene.

<p>Ref on the alignment indicates the nucleotide sequences of the Ov 7 gene of the OvHV-2 reference genomes obtained from the GenBank. Variable regions are highlighted in rectangles (▯) and group of genotypes (G) are illustrated on the right side of the align sequences.</p

Multiple sequence alignment of amino acid sequences of the ORF 73 gene from SA OvHV-2 strains and reference sequences derived from OvHV-2 genome sequences (Accession numbers: NC-007646.1, AY839756.1, DQ198083.1) analysed with ExPASy translate tool.

<p>Ref indicates the amino acid sequences of the ORF 73 gene of the OvHV-2 reference genomes obtained from the GenBank. Variable regions are highlighted in rectangles (▯), and groups of amino acid sequences (G) are illustrated on the right side of the align sequences</p

The list of bovine blood samples, obtained from different geographic areas in South Africa, used for molecular characterization of OvHV-2.

<p>The list of bovine blood samples, obtained from different geographic areas in South Africa, used for molecular characterization of OvHV-2.</p

The phylogenetic trees of the ORF 27 nucleotide (A & B) and amino acid (C & D) sequences of OvHV-2 strains and their closely related reference sequences obtained from GenBank constructed using Neighbor-Joining (A & C) and Maximum Likelihood (B & D) methods.

<p>The unrooted condensed trees were constructed using Neighbor-Joining (A & C) and Maximum Likelihood (B & D) methods, showing branching patterns of each clade. Groups 1 and 2 are illustrated and abbreviated as clade 1 (C1) and 2 (C2), respectively, with supporting bootstrap values at each clade nodes.</p

The phylogenetic trees of the Ov 7 nucleotide (A & B) and amino acid (C & D) sequences of OvHV-2 strains and their closely related reference sequences obtained from GenBank.

<p>The unrooted condensed trees were constructed using Neighbor-Joining (A & C) and Maximum Likelihood (B & D) methods, showing branching patterns of each clade. Groups 1, 2 and 3 are illustrated and abbreviated as clade 1 (C1), 2 (C2) and 3 C3), respectively, with supporting bootstrap values at each clade nodes.</p