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. 2019 Mar 8;15(1):84.
doi: 10.1186/s12917-019-1828-6.

Molecular detection and phylogenetic analysis of Peste des petits ruminants virus circulating in small ruminants in eastern Amhara region, Ethiopia

Biruk Alemu  1 , Getachew Gari  2 , Geneviève Libeau  3 , Olivier Kwiatek  3 , Menbere Kidane  4 , Rediet Belayneh  4 , Bewuket Siraw  5 , Barbara Wieland  6 , Wondwoson Asfaw  7 , Reta D Abdi  8   9
Affiliations

Molecular detection and phylogenetic analysis of Peste des petits ruminants virus circulating in small ruminants in eastern Amhara region, Ethiopia

Biruk Alemu et al. BMC Vet Res. .

Abstract

Background: Peste des Petits Ruminants (PPR) is a severe, highly infectious and fatal viral disease of small ruminants. Four lineages of PPR virus have been identified globally based on sequence analysis of the nucleoprotein (N) and fusion (F) gene. The aim of this study was to isolate and genetically characterize recently circulating PPR virus in small ruminants in the eastern Amhara region in Ethiopia. A total of 28 anti-mortem samples (gum debris, nasal and ocular swab) were collected from clinically suspicious animals and examined for the presence of PPRV by a one-step RT-PCR assay. Samples positive with RT-PCR were subjected to isolation of the virus which were subsequently genetically characterized by sequencing of the nucleoprotein (N) gene and phylogenetic analysis of PPR virus (PPRV) strains.

Results: Of the 28 clinical samples examined, 46.4% were positive with RT-PCR for viral nucleic acid. The PPRV was successfully isolated on CHS-20 cell line with the ovine signaling lymphocyte activation molecule (SLAM) receptor expressed on the cell surface and confirmed with RT-PCR and IFAT assay. The nucleotide sequence and phylogenetic analysis indicated that the PPRV obtained were clustered genetically with Lineage IV isolates of the virus.

Conclusion: The successful isolation of the virus and molecular findings of this study confirmed active lineage IV PPRV infections among populations of sheep and goats in eastern Amhara, suggesting risks for potential spread of the disease to currently free areas. Thus, we recommend systematic vaccination to contain outbreaks in affected districts and geographically linked surrounding districts to which the disease could potentially spread due to different epidemiological linkages.

Keywords: Eastern Amhara; Isolation; Molecular characterization; PPRV; Small ruminants.

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Conflict of interest statement

Ethics approval and consent to participate

This study was approved by the animal research ethical review committee of Addis Ababa University (Certificate ref. No: VM/ERC/02/05/10/2018) and we received oral consent from the animal owner to take clinical samples from their animals.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Map of Ethiopia showing regions, study zones, districts and sampling sites. PA: peasant association names used by local community; PAs2007: peasant association names sourced from 2007 census map
Fig. 2
Fig. 2
Observed clinical signs of PPR: a) Erosive and necrotic stomatitis, b) the upper dental pad completely hidden by a thick cheese-like material, c) Ulceration on the upper surface of the tongue, d) serious nasal discharge, dead cells on the surface of tongue and lesion on lower lip, e) muco-purulent nasal discharge, and f) lacrimation
Fig. 3
Fig. 3
Agarose gel electrophoresis of PCR products (351 bp) amplified with NP3 and NP4, PPR specific primers. Lane M: 100 bp DNA molecular weight marker; Lane P: Positive control; Lane N: Negative control; Lane 1–11: Field samples
Fig. 4
Fig. 4
Phylogenetic analysis of nucleotide sequences from the amplified products of PPRV N protein gene with different lineages occurring worldwide
See this image and copyright information in PMC

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