Paramyxoviruses, negative-sense single-stranded RNA viruses, pose a critical threat to human public health. Currently, 78 species, 17 genera, and 4 subfamilies of paramyxoviruses are harbored by multiple natural reservoirs, including rodents, bats, birds, reptiles, and fish. Henipaviruses are critical zoonotic pathogens that cause severe acute respiratory distress and neurological diseases in humans. Using reverse transcription-polymerase chain reaction, 115 Crocidura species individuals were examined for the prevalence of paramyxovirus infections. Paramyxovirus RNA was observed in 26 (22.6%) shrews collected at five trapping sites, Republic of Korea. Herein, we report two genetically distinct novel paramyxoviruses (genus: Henipavirus): Gamak virus (GAKV) and Daeryong virus (DARV) isolated from C. lasiura and C. shantungensis, respectively. Two GAKVs and one DARV were nearly completely sequenced using next-generation sequencing. GAKV and DARV contain six genes (3’-N-P-M-F-G-L-5´) with genome sizes of 18,460 nucleotides and 19,471 nucleotides, respectively. The phylogenetic inference demonstrated that GAKV and DARV form independent genetic lineages of Henipavirus in Crocidura species. GAKV-infected human lung epithelial cells elicited the induction of type I/III interferons, interferon-stimulated genes, and proinflammatory cytokines. In conclusion, this study contributes further understandings of the molecular prevalence, genetic characteristics and diversity, and zoonotic potential of novel paramyxoviruses in shrews.
Actinomycetes are Gram positive, free living saprophytes which are distributed in soil as one of the major populations and are primary source of antibiotics. This study was carried out with a quest to isolate actinomycetes from soil samples of different places and assess their antibacterial activity. Isolation of actinomycetes was carried out by serial dilution of soil sample followed by spread plate method. The antimicrobial extract was extracted using ethyl acetate. Assessment of antimicrobial activity was performed by using Agar cup plate assay against test organisms (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Salmonella paratyphi, Bacillus subtilis, Staphylococcus aureus). Antibacterial activity was tested against Methicillin Sensitive Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus in the isolates having effective inhibitory activity against Staphylococcus aureus. From 15 soil samples of 12 different locations, 121 actinomycetes isolates were isolated. Among them, 58 (47.9%) isolates were inhibitory against at least 1 test organism in primary screening, of which 22 isolates effective against more than 1 test organism was chosen for secondary screening. Out of them, 8 were inhibitory against 2 test organisms while 14 were inhibitory against 3 test organisms. Staphylococcus aureus was found to be the most susceptible test organism with its susceptibility against 12 actinomycetes isolates. Among 12 isolates effective against Staphylococcus aureus, 10 were found to have an inhibitory effect against Methicillin Susceptible Staphylococcus aureus while 6 were found to have inhibitory effect against Methicillin Resistant Staphylococcus aureus strain. The findings of this study highlight the inhibitory potential of actinomycetes and the need for further investigation for obtaining novel antimicrobial agents from actinomycetes from various unexplored areas.
Whole-genome sequencing of infectious agents enables the identification and characterization of emerging viruses. The MinION device is a portable sequencer that allows real-time sequencing in fields or hospitals. Hantaan orthohantavirus (Hantaan virus, HTNV), harbored by Apodemus agrarius, causes hemorrhagic fever with renal syndrome (HFRS) and poses a critical public health threat worldwide. In this study, we aimed to evaluate the feasibility of using nanopore sequencing for whole-genome sequencing of HTNV from samples having different viral copy numbers. Amplicon-based next-generation sequencing was performed in A. agrarius lung tissues collected from the Republic of Korea. Genomic sequences of HTNV were analyzed based on the viral RNA copy numbers. Amplicon-based nanopore sequencing provided nearly full-length genomic sequences of HTNV and showed sufficient read depth for phylogenetic analysis after 8 h of sequencing. The average identity of the HTNV genome sequences for the nanopore sequencer compared to those of generated from Illumina MiSeq revealed 99.8% (L and M segments) and 99.7% (S segment) identities, respectively. This study highlights the potential of the portable nanopore sequencer for rapid generation of accurate genomic sequences of HTNV for quicker decision making in point-of-care testing of HFRS patients during a hantavirus outbreak.
Paramyxoviruses, negative-sense single-stranded RNA viruses, pose a potential threat to public health. Currently, 78 species and 17 genera of paramyxoviruses are classified and harbored by multiple natural reservoirs, including rodents, bats, birds, reptiles, and fish. Jeilongvirus has been proposed as a novel paramyxovirus genus containing J-, Beilong, and Tailam viruses, found in wild rodents. Using RT-PCR, 824 Apodemus agrarius individuals were examined for the prevalence of paramyxovirus infections. Paramyxovirus RNA was detected in 108 (13.1%) rodents captured at 14 trapping sites in Korea. We first present two genetically distinct novel paramyxoviruses (genus Jeilongvirus), Paju Apodemus paramyxoviruses 1 (PAPV-1) and 2 (PAPV-2), from A. agrarius. Six PAPV strains were completely sequenced using next-generation and Sanger sequencing. PAPV-1 genome comprised 19,716 nucleotides, with eight genes (3-N-P/V/C-M-F-SH-TM-G-L-5), whereas PAPV-2 genome contained 17,475 nucleotides, with seven genes (3-N-P/V/C-M-F-TM-G-L-5). The disparity between PAPV-1 and -2 revealed the presence of the SH gene and length of the G gene in the genome organization. The phylogenies of PAPV-1 and -2 belong to distinct genetic lineages of Jeilongvirus despite being from the same natural host. PAPV-1 clustered with Beilong and Tailam viruses, while PAPV-2 formed a genetic lineage with Mount Mabu Lophuromys virus-1. PAPV-1 infected human epithelial and endothelial cells, facilitating the induction of type I/III interferons, interferon-stimulated genes, and proinflammatory cytokines. Therefore, this study provides profound insights into the molecular epidemiology, virus-host interactions, and zoonotic potential of novel rodent-borne paramyxoviruses.
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