Abstract

Identification of unknown pathogens in pigs displaying enteric illness is difficult due to the large diversity of bacterial and viral species found within faecal samples. Current methods often require bacterial or viral isolation, or testing only a limited number of known species using quantitative PCR analysis. Herein, faeces from two 25-day-old piglets with diarrhoea from Texas, USA, were analysed by metagenomic next-generation sequencing to rapidly identify possible pathogens. Our analysis included a bioinformatics pipeline of rapid short-read classification and de novo genome assembly which resulted in the identification of a porcine enterovirus G (EV-G), a complete genome with substantial nucleotide differences (>30 %) among current sequences, and a novel non-structural protein similar in sequence to the Torovirus papain-like cysteine protease (PLpro). This discovery led to the identification and circulation of an EV-G with a novel PLpro in the USA that has not been previously reported.

Highlights

  • Identification of unknown pathogens in pigs displaying enteric illness is difficult due to the large diversity of bacterial and viral species found within faecal samples

  • Molecular subtyping based on VP1, VP4/2 or 3D sequence alignments have revealed a large diversity of enterovirus G (EV-G) genotypes

  • The assembly and verification pipeline generated accurate and complete EV-G genomes for both samples and the strain sequences were submitted to GenBank (EVG/Porcine/USA/Texas1/2014/G1-papain-like cysteine protease (PLpro) and EVG/Porcine/USA/ Texas2/2014/G1-PLpro, accession numbers: KY498016 and KY498017, respectively)

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Summary

Introduction

Identification of unknown pathogens in pigs displaying enteric illness is difficult due to the large diversity of bacterial and viral species found within faecal samples. This workflow identified and assembled an enterovirus G complete genome that contained a novel 669 nucleotide insertion from two diarrhoeic 25-day-old piglets at weaning from Texas, USA.

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