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Molecular characterization of porcine group A rotavirus to contain piglet diarrhea for productivity enhancement in North East India.

ABSTRACT: Gastrointestinal tract infections with acute diarrhea are indeed a serious cause of concern that held back the swine industry of North East India and among the various etiological agents causing the disease, group A rotavirus is the most common. In this regard, molecular characterization of the viral strains circulating in the region is of utmost importance for a strategic approach to control the disease. Thus, in the present study, diarrheic fecal samples from piglets of Tripura and Assam were evaluated for genotypic characterization of VP7 and VP4 gene of the virus. The samples positive for group A rotavirus were subjected to G and P type determination by nested- multiplex RT PCR as well as nucleotide sequencing of VP7 and VP4 gene. Nested- multiplex RT PCR revealed the presence of porcine group A rotavirus in the genotypic combination of G1P[6], G3P[6], G8P[6] and G4P[6] in the two states of North East India. Nucleotide sequencing of VP7 and VP4 genes however, revealed emergence of an unusual genotypic combination G26P[13]. VP7 and VP4 nucleotide and its deduced amino acid sequences when compared with global isolates, showed that the present strains shared closest similarity to the porcine rotavirus strain TJ4-1from Japan and hp140 from Eastern India. Identification of a novel genotype G26P[13] in the RVA infected pig population of Tripura and Assam, suggested presence of a greater diversity in rotavirus strains in the region. Thus, it evokes the need for continuous monitoring to assess and consider the strain variability in the design of a suitable vaccine candidate against the virus.

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The online version of this article (10.1007/s13337-021-00659-6) contains supplementary material, which is available to authorized users.

SUBMITTER: Das S

PROVIDER: S-EPMC8035605 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus that causes high mortality in piglets, thus posing a serious threat to the world pig industry. Porcine epidemic diarrhea (PED) is related to the imbalance of sodium absorption by small intestinal epithelial cells; however, the etiology of sodium imbalanced diarrhea caused by PEDV remains unclear. Herein, we first proved that PEDV can cause a significant decrease in Na+/H+ exchanger 3 (NHE3) expression on the cell membrane, in a viral dose-dependent manner. Further study showed that the PEDV nucleocapsid (N) protein participates in the regulation of NHE3 activity through interacting with Ezrin. Flame atomic absorption spectroscopy results indicated a serious imbalance in Na+ concentration inside and outside cells following overexpression of PEDV N. Meanwhile, molecular docking technology identified that the small molecule drug Pemetrexed acts on the PEDV N-Ezrin interaction region. It was confirmed that Pemetrexed can alleviate the imbalanced Na+ concentration in IPEC-J2 cells and the diarrhea symptoms of Rongchang pigs caused by PEDV infection. Overall, our data suggest that the interaction between PEDV N and Ezrin reduces the level of phosphorylated Ezrin, resulting in a decrease in the amount of NHE3 protein on the cell membrane. This leads to an imbalance of intracellular and extracellular Na+, which causes diarrhea symptoms in piglets. Pemetrexed is effective in relieving diarrhea caused by PEDV. Our results provide a reference to screen for anti-PEDV targets and to develop drugs to prevent PED.IMPORTANCEPorcine epidemic diarrhea (PED) has caused significant economic losses to the pig industry since its initial outbreak, and the pathogenic mechanism of porcine epidemic diarrhea virus (PEDV) is still under investigation. Herein, we found that the PEDV nucleocapsid protein interacts with Ezrin to regulate Na+/H+ exchanger 3 activity. In addition, we screened out Pemetrexed, a small molecule drug, which can effectively alleviate pig diarrhea caused by PEDV. These results provide support for further exploration of the pathogenesis of PEDV and the development of drugs to prevent PED.

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Molecular characterization of porcine group A rotavirus to contain piglet diarrhea for productivity enhancement in North East India.

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