Orthoreoviruses infect virtually all mammalian species, causing systemic infections including mild gastrointestinal and respiratory illnesses. However, little is known about the prevalence or genetic diversity of porcine orthoreoviruses in South Korea. We examined 237 diarrheic fecal samples collect...
Orthoreoviruses infect virtually all mammalian species, causing systemic infections including mild gastrointestinal and respiratory illnesses. However, little is known about the prevalence or genetic diversity of porcine orthoreoviruses in South Korea. We examined 237 diarrheic fecal samples collected from 78 pig farms around the country. RT-PCR utilizing primers specific for the L1 gene of mammalian orthoreoviruses showed that 45 (19.0%) samples were positive. The 10 strains isolated from orthoreovirus-positive samples formed typical perinuclear cytoplasmic inclusion bodies and had an atypical hemagglutination pattern; these are characteristics of type 3 orthoreovirus. Phylogenetic analysis of the S1 gene in these 10 Korean and other strains showed that type 3 orthoreoviruses could be divided into four lineages; the 10 Korean strains were included in porcine lineage IV, along with T3/porcine/Sichuan/2006. Sequence analysis showed that strains in lineage IV had nucleotide identities of 97.0-98.1% and deduced amino acid identities of 96.4-98.2%. Sequence analysis of the σ1 protein, a viral attachment protein, revealed that the amino acid sequences associated with neurotropism (amino acids 198-204, 249I, 350D, and 419E) were highly conserved among the Korean strains, confirming that neural tropism was present. In conclusion, our findings suggest that porcine orthoreovirus infections are endemic in pig farms in South Korea and that the 10 novel Korean porcine orthoreoviruses belong to porcine lineage IV of type 3 orthoreovirus. In addition, sequence analysis of S1 genes encoding the σ1 protein showed that the Korean porcine orthoreoviruses exhibited neural tropism.
In this study, colostrums-deprived piglets were experimentally inoculated with the Korean KPR113 strain and examined for viremia, enteric and nasal virus shedding as well as for viral antigen expression and virus associated lesions in the small and large intestines, extra-intestinal spread in piglets. The KPR113 strain caused severe diarrhea in experimentally infected piglets with gradual villous atrophy in the small intestine and a gradual increase in the crypt depth of the large intestine. The KPR113 strain inoculated piglets showed epithelial damage in nasal turbinates, trachea and lung, and interstitial pneumonia. By SYBR Green real-time PCR, viral RNA was detected in feces of the piglets at post-inoculation day (PID) 1, reaching a peak at PID5, and then rapidly decreasing from PID7. In addition serum, mesenteric lymph node, lungs, liver, choroid plexus. These results show that the KPR113 strain has dual tropism and induces pathological changes in both the digestive and respiratory tracts of piglets.
We evaluated the ability of MeOH extract and ethyl acetate fraction, kuraridin compound from the Shphora flavescens to porcine reovirus. we assessed the anti-viral activity of pre-treatment, simultaneous treatment, and post treatment. Using the simultaneous treatment assay, we found that extracts and fraction and compounds directly blocking viral adsorption to cells. Kuraridin inhibited T1L, T2J, T3D, KPR 113 reovirus absorption with EC50 values of 41.1, 176.9, 26.7, 15.3 μM, respectively. Moreover, the post treatment assay, showed that kuraridin inhibited T1L, T2J, T3D, KPR 113 reovirus viral replication with EC50 values of 62.0, 29.4, 14.4, 14.0 μM, respectively. By the hemagglutination inhibition (HI) assay, one MeOH extracts, EA fraction and one compounds completely inhibited viral adsorption onto bovine RBCs at less than 40 μg/mL or 20 μM against KPR 113 strain. Especially, kuraridin was found with inhibition for both viral attachment and viral replication after showing extended antiviral activity during the quantitative real-time PCR.
Orthoreoviruses infect virtually all mammalian species, causing systemic infections including mild gastrointestinal and respiratory illnesses. However, little is known about the prevalence or genetic diversity of porcine orthoreoviruses in South Korea. We examined 237 diarrheic fecal samples collected from 78 pig farms around the country. RT-PCR utilizing primers specific for the L1 gene of mammalian orthoreoviruses showed that 45 (19.0%) samples were positive. The 10 strains isolated from orthoreovirus-positive samples formed typical perinuclear cytoplasmic inclusion bodies and had an atypical hemagglutination pattern; these are characteristics of type 3 orthoreovirus. Phylogenetic analysis of the S1 gene in these 10 Korean and other strains showed that type 3 orthoreoviruses could be divided into four lineages; the 10 Korean strains were included in porcine lineage IV, along with T3/porcine/Sichuan/2006. Sequence analysis showed that strains in lineage IV had nucleotide identities of 97.0-98.1% and deduced amino acid identities of 96.4-98.2%. Sequence analysis of the σ1 protein, a viral attachment protein, revealed that the amino acid sequences associated with neurotropism (amino acids 198-204, 249I, 350D, and 419E) were highly conserved among the Korean strains, confirming that neural tropism was present. In conclusion, our findings suggest that porcine orthoreovirus infections are endemic in pig farms in South Korea and that the 10 novel Korean porcine orthoreoviruses belong to porcine lineage IV of type 3 orthoreovirus. In addition, sequence analysis of S1 genes encoding the σ1 protein showed that the Korean porcine orthoreoviruses exhibited neural tropism.
In this study, colostrums-deprived piglets were experimentally inoculated with the Korean KPR113 strain and examined for viremia, enteric and nasal virus shedding as well as for viral antigen expression and virus associated lesions in the small and large intestines, extra-intestinal spread in piglets. The KPR113 strain caused severe diarrhea in experimentally infected piglets with gradual villous atrophy in the small intestine and a gradual increase in the crypt depth of the large intestine. The KPR113 strain inoculated piglets showed epithelial damage in nasal turbinates, trachea and lung, and interstitial pneumonia. By SYBR Green real-time PCR, viral RNA was detected in feces of the piglets at post-inoculation day (PID) 1, reaching a peak at PID5, and then rapidly decreasing from PID7. In addition serum, mesenteric lymph node, lungs, liver, choroid plexus. These results show that the KPR113 strain has dual tropism and induces pathological changes in both the digestive and respiratory tracts of piglets.
We evaluated the ability of MeOH extract and ethyl acetate fraction, kuraridin compound from the Shphora flavescens to porcine reovirus. we assessed the anti-viral activity of pre-treatment, simultaneous treatment, and post treatment. Using the simultaneous treatment assay, we found that extracts and fraction and compounds directly blocking viral adsorption to cells. Kuraridin inhibited T1L, T2J, T3D, KPR 113 reovirus absorption with EC50 values of 41.1, 176.9, 26.7, 15.3 μM, respectively. Moreover, the post treatment assay, showed that kuraridin inhibited T1L, T2J, T3D, KPR 113 reovirus viral replication with EC50 values of 62.0, 29.4, 14.4, 14.0 μM, respectively. By the hemagglutination inhibition (HI) assay, one MeOH extracts, EA fraction and one compounds completely inhibited viral adsorption onto bovine RBCs at less than 40 μg/mL or 20 μM against KPR 113 strain. Especially, kuraridin was found with inhibition for both viral attachment and viral replication after showing extended antiviral activity during the quantitative real-time PCR.
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