#Paramyxo- and #Coronaviruses in #Rwandan #Bats (Trop Med Infect Dis., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Trop Med Infect Dis. 2019 Jul 2;4(3). pii: E99. doi: 10.3390/tropicalmed4030099.

Paramyxo- and Coronaviruses in Rwandan Bats.

Markotter W1, Geldenhuys M2, Jansen van Vuren P2,3, Kemp A3, Mortlock M2, Mudakikwa A4, Nel L5, Nziza J6, Paweska J2,3, Weyer J2,3.

Author information: 1 Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria, Gauteng 0001, South Africa. wanda.markotter@up.ac.za. 2 Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria, Gauteng 0001, South Africa. 3 Centre for Emerging Zoonotic and Parasitic diseases, National Institute for Communicable Diseases, National Health laboratory Services, Sandringham, Johannesburg 2131, South Africa. 4 Rwanda Development Board, Department of tourism and Conservation, P.O Box 6239, Kigali, Rwanda. 5 Centre for Viral Zoonoses, Department of Biochemistry, Genetics and Microbiology, Faculty of Natural and Agricultural Sciences, University of Pretoria, Pretoria, Gauteng 0001, South Africa. 6 Mountain Gorilla Veterinary Project, P.O Box 115, Musanze, Rwanda.

 

Abstract

A high diversity of corona- and paramyxoviruses have been detected in different bat species at study sites worldwide, including Africa, however no biosurveillance studies from Rwanda have been reported. In this study, samples from bats collected from caves in Ruhengeri, Rwanda, were tested for the presence of corona- and paramyxoviral RNA using reverse transcription PCR assays. Positive results were further characterized by DNA sequencing and phylogenetic analysis. In addition to morphological identification of bat species, we also did molecular confirmation of species identities, contributing to the known genetic database available for African bat species. We detected a novel Betacoronavirus in two Geoffroy’s horseshoe bats (Rhinolophus clivosus) bats. We also detected several different paramyxoviral species from various insectivorous bats. One of these viral species was found to be homologous to the genomes of viruses belonging to the Jeilongvirus genus. Additionally, a Henipavirus-related sequence was detected in an Egyptian rousette fruit bat (Rousettus aegyptiacus). These results expand on the known diversity of corona- and paramyxoviruses and their geographical distribution in Africa.

KEYWORDS: Rwanda; barcoding; bat; caves; coronavirus; henipavirus; jeilongvirus; paramyxovirus; surveillance

PMID: 31269631 DOI: 10.3390/tropicalmed4030099

Keywords: Coronavirus; Betacoronavirus; Paramyxovirus; Henipavirus; Bats; Rwanda.

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#Human #coronavirus alone or in co-infection with #rhinovirus C is a #risk factor for #SARI and admission to the #PICU: A one-year study in Southeast #Brazil (PLoS One, abstract)

[Source: PLoS One, full page: (LINK). Abstract, edited.]

OPEN ACCESS /  PEER-REVIEWED / RESEARCH ARTICLE

Human coronavirus alone or in co-infection with rhinovirus C is a risk factor for severe respiratory disease and admission to the pediatric intensive care unit: A one-year study in Southeast Brazil

Alessandra K. Matsuno  , Talita B. Gagliardi , Flavia E. Paula, Luciano K. S. Luna, Bruna L. S. Jesus, Renato T. Stein, Davi C. Aragon, Ana P. C. P. Carlotti , Eurico Arruda

Published: June 3, 2019 / DOI: https://doi.org/10.1371/journal.pone.0217744

 

Abstract

Objective

We aimed to assess the profile of respiratory viruses in young children hospitalized for acute lower respiratory tract infection (ALRI) and its association with disease severity, defined as need for pediatric intensive care unit (PICU) admission.

Design

Prospective observational cohort study.

Setting

A tertiary-care university hospital in Brazil.

Patients

Children younger than three years attending the pediatric emergency room with ALRI who were admitted to the hospital.

Interventions

None.

Measurements and main results

Nasopharyngeal aspirates were collected from patients from June 1st, 2008 to May 31st, 2009within the first 48 hours of hospitalization. Nasopharyngeal aspirates were tested for 17 human respiratory viruses by molecular and immunofluorescence based assays. Simple and multiple log-binomial regression models were constructed to assess associations of virus type with a need for PICU admission. Age, prematurity, the presence of an underlying disease and congenital heart disease were covariates. Nasopharyngeal aspirates were positive for at least one virus in 236 patients. Rhinoviruses were detected in 85.6% of samples, with a preponderance of rhinovirus C (RV-C) (61.9%). Respiratory syncytial virus was detected in 59.8% and human coronavirus (HCoV) in 11% of the samples. Co-detections of two to five viruses were found in 78% of the patients. The detection of HCoV alone (adjusted relative risk (RR) 2.18; 95% CI 1.15–4.15) or in co-infection with RV-C (adjusted RR 2.37; 95% CI 1.23–4.58) was independently associated with PICU admission.

Conclusions

The detection of HCoV alone or in co-infection with RV-C was independently associated with PICU admission in young children hospitalized for ALRI.

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Citation: Matsuno AK, Gagliardi TB, Paula FE, Luna LKS, Jesus BLS, Stein RT, et al. (2019) Human coronavirus alone or in co-infection with rhinovirus C is a risk factor for severe respiratory disease and admission to the pediatric intensive care unit: A one-year study in Southeast Brazil. PLoS ONE 14(6): e0217744. https://doi.org/10.1371/journal.pone.0217744

Editor: Raymond J. Pickles, University of North Carolina at Chapel Hill, UNITED STATES

Received: October 2, 2018; Accepted: May 19, 2019; Published: June 3, 2019

Copyright: © 2019 Matsuno et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the manuscript and its Supporting Information files.

Funding: AKM and EAN supported mainly by an academic grant from Abbott Laboratórios do Brasil Ltda based on an unrestricted investigator-generated proposal. EAN Additional funding was provided by a grant from the Sao Paulo State Research Foundation (FAPESP) and by FAEPA – “Fundação de Apoio ao Ensino Pesquisa e Assistência” (in English would be “Foundation of Support to Teaching Research and Assistance) of the University of Sao Paulo Hospital of Ribeirão Preto. EA has long standing scholarship support from the Brazilian Research Council (CNPq). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: AKM and EAN were supported by an academic grant from Abbott Laboratórios do Brasil Ltda, based on an unrestricted investigator-generated proposal. This does not alter our adherence to PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors: https://journals.plos.org/plosone/s/competing-interests).

Keywords: Coronavirus; Rhinovirus; SARI; Intensive Care; Pediatrics; Brazil.

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High-Throughput #Screening and Identification of Potent Broad-Spectrum #Inhibitors of #Coronaviruses (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses

Liang Shen, Junwei Niu, Chunhua Wang, Baoying Huang, Wenling Wang, Na Zhu, Yao Deng, Huijuan Wang, Fei Ye, Shan Cen, Wenjie Tan

Tom Gallagher, Editor

DOI: 10.1128/JVI.00023-19

 

ABSTRACT

Coronaviruses (CoVs) act as cross-species viruses and have the potential to spread rapidly into new host species and cause epidemic diseases. Despite the severe public health threat of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome CoV (MERS-CoV), there are currently no drugs available for their treatment; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are urgently needed. To search for effective inhibitory agents, we performed high-throughput screening (HTS) of a 2,000-compound library of approved drugs and pharmacologically active compounds using the established genetically engineered human CoV OC43 (HCoV-OC43) strain expressing Renilla luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs in vitro. We screened 56 hits from the HTS data and validated 36 compounds in vitro using wild-type HCoV-OC43. Furthermore, we identified seven compounds (lycorine, emetine, monensin sodium, mycophenolate mofetil, mycophenolic acid, phenazopyridine, and pyrvinium pamoate) as broad-spectrum inhibitors according to their strong inhibition of replication by four CoVs in vitroat low-micromolar concentrations. Additionally, we found that emetine blocked MERS-CoV entry according to pseudovirus entry assays and that lycorine protected BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This represents the first demonstration of in vivo real-time bioluminescence imaging to monitor the effect of lycorine on the spread and distribution of HCoV-OC43 in a mouse model. These results offer critical information supporting the development of an effective therapeutic strategy against CoV infection.

 

IMPORTANCE

Currently, there is no approved therapy to treat coronavirus infection; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are needed. Based on our high-throughput screening assay using a compound library, we identified seven compounds with broad-spectrum efficacy against the replication of four CoVs in vitro. Additionally, one compound (lycorine) was found to protect BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This inhibitor might offer promising therapeutic possibilities for combatting novel CoV infections in the future.

 

FOOTNOTES

Received 7 January 2019. Accepted 17 March 2019. Accepted manuscript posted online 27 March 2019.

Supplemental material for this article may be found at https://doi.org/10.1128/JVI.00023-19.

Copyright © 2019 American Society for Microbiology. All Rights Reserved.

Keywords: Antivirals; Coronavirus.

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Analysis of #Coronavirus Temperature-Sensitive Mutants Reveals an Interplay between the Macrodomain and Papain-Like Protease Impacting #Replication and #Pathogenesis (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Analysis of Coronavirus Temperature-Sensitive Mutants Reveals an Interplay between the Macrodomain and Papain-Like Protease Impacting Replication and Pathogenesis

Xufang Deng, Robert C. Mettelman, Amornrat O’Brien, John A. Thompson, Timothy E. O’Brien, Susan C. Baker

Julie K. Pfeiffer, Editor

DOI: 10.1128/JVI.02140-18

 

ABSTRACT

Analysis of temperature-sensitive (ts) mutant viruses is a classic method allowing researchers to identify genetic loci involved in viral replication and pathogenesis. Here, we report genetic analysis of a ts strain of mouse hepatitis virus (MHV), tsNC11, focusing on the role of mutations in the macrodomain (MAC) and the papain-like protease 2 (PLP2) domain of nonstructural protein 3 (nsp3), a component of the viral replication complex. Using MHV reverse genetics, we generated a series of mutant viruses to define the contributions of macrodomain- and PLP2-specific mutations to the ts phenotype. Viral replication kinetics and efficiency-of-plating analysis performed at permissive and nonpermissive temperatures revealed that changes in the macrodomain alone were both necessary and sufficient for the ts phenotype. Interestingly, mutations in the PLP2 domain were not responsible for the temperature sensitivity but did reduce the frequency of reversion of macrodomain mutants. Coimmunoprecipitation studies are consistent with an interaction between the macrodomain and PLP2. Expression studies of the macrodomain-PLP2 portion of nsp3 indicate that the ts mutations enhance proteasome-mediated degradation of the protein. Furthermore, we found that during virus infection, the replicase proteins containing the MAC and PLP2 mutations were more rapidly degraded at the nonpermissive temperature than were the wild-type proteins. Importantly, we show that the macrodomain and PLP2 mutant viruses trigger production of type I interferon in vitro and are attenuated in mice, further highlighting the importance of the macrodomain-PLP2 interplay in viral pathogenesis.

 

IMPORTANCE

Coronaviruses (CoVs) are emerging human and veterinary pathogens with pandemic potential. Despite the established and predicted threat these viruses pose to human health, there are currently no approved countermeasures to control infections with these viruses in humans. Viral macrodomains, enzymes that remove posttranslational ADP-ribosylation of proteins, and viral multifunctional papain-like proteases, enzymes that cleave polyproteins and remove polyubiquitin chains via deubiquitinating activity, are two important virulence factors. Here, we reveal an unanticipated interplay between the macrodomain and the PLP2 domain that is important for replication and antagonizing the host innate immune response. Targeting the interaction of these enzymes may provide new therapeutic opportunities to treat CoV disease.

Copyright © 2019 American Society for Microbiology. All Rights Reserved.

Keywords: Coronavirus; Viral pathogenesis.

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#Nucleoside analogues for the #treatment of #coronavirus #infections (Curr Opin Virol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Curr Opin Virol. 2019 May 21;35:57-62. doi: 10.1016/j.coviro.2019.04.002. [Epub ahead of print]

Nucleoside analogues for the treatment of coronavirus infections.

Pruijssers AJ1, Denison MR2.

Author information: 1 Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, TN, United States; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, TN, United States. 2 Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, TN, United States; Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, TN, United States; Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, TN, United States. Electronic address: mark.denison@vumc.org.

 

Abstract

Recent outbreaks of SARS-Coronavirus and MERS-Coronavirus (CoV) have heightened awareness about the lack of vaccines or antiviral compounds approved for prevention or treatment of human or potential zoonotic CoVs. Anti-CoV drug development has long been challenged by the activity of a 3′ to 5′ proofreading exoribonuclease unique to CoVs. Recently, a promising nucleoside analogue with broad-spectrum activity against CoVs has been identified. This review will discuss progress made in the development of antiviral nucleoside and nucleotide analogues targeting viral RNA synthesis as effective therapeutics against CoV infections and propose promising strategies for combination therapy.

Copyright © 2019. Published by Elsevier B.V.

PMID: 31125806 DOI: 10.1016/j.coviro.2019.04.002

Keywords: Coronavirus; MERS-CoV; Antivirals.

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Complete #genome analysis of a #SARS-like #bat #coronavirus identified in the Republic of #Korea (Virus Genes, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Virus Genes. 2019 May 10. doi: 10.1007/s11262-019-01668-w. [Epub ahead of print]

Complete genome analysis of a SARS-like bat coronavirus identified in the Republic of Korea.

Kim Y1,2, Son K1, Kim YS2, Lee SY2, Jheong W1, Oem JK3.

Author information: 1 Environmental Health Research Department, National Institute of Environmental Research, Hwangyeong-ro 42, Seo-gu, Incheon, Republic of Korea. 2 Department of Veterinary Infectious Diseases, College of Veterinary Medicine, Chonbuk National University, Jeonju, Republic of Korea. 3 Department of Veterinary Infectious Diseases, College of Veterinary Medicine, Chonbuk National University, Jeonju, Republic of Korea. jku0623@jbnu.ac.kr.

 

Abstract

Bats have been widely known as natural reservoir hosts of zoonotic diseases, such as severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) caused by coronaviruses (CoVs). In the present study, we investigated the whole genomic sequence of a SARS-like bat CoV (16BO133) and found it to be 29,075 nt in length with a 40.9% G+C content. Phylogenetic analysis using amino acid sequences of the ORF 1ab and the spike gene showed that the bat coronavirus strain 16BO133 was grouped with the Beta-CoV lineage B and was closely related to the JTMC15 strain isolated from Rhinolophus ferrumequinum in China. However, 16BO133 was distinctly located in the phylogenetic topology of the human SARS CoV strain (Tor2). Interestingly, 16BO133 showed complete elimination of ORF8 regions induced by a frame shift of the stop codon in ORF7b. The lowest amino acid identity of 16BO133 was identified at the spike region among various ORFs. The spike region of 16BO133 showed 84.7% and 75.2% amino acid identity with Rf1 (SARS-like bat CoV) and Tor2 (human SARS CoV), respectively. In addition, the S gene of 16BO133 was found to contain the amino acid substitution of two critical residues (N479S and T487 V) associated with human infection. In conclusion, we firstly carried out whole genome characterization of the SARS-like bat coronavirus discovered in the Republic of Korea; however, it presumably has no human infectivity. However, continuous surveillance and genomic characterization of coronaviruses from bats are necessary due to potential risks of human infection induced by genetic mutation.

KEYWORDS: Bat; Frame shift; SARS-like coronavirus; Whole genome; Zoonotic disease

PMID: 31076983 DOI: 10.1007/s11262-019-01668-w

Keywords: Coronavirus; SARS; Bats; S. Korea.

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#Discovery and Characterization of Novel #Bat #Coronavirus Lineages from #Kazakhstan (Viruses, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Viruses. 2019 Apr 17;11(4). pii: E356. doi: 10.3390/v11040356.

Discovery and Characterization of Novel Bat Coronavirus Lineages from Kazakhstan.

Mendenhall IH1, Kerimbayev AA2, Strochkov VM3, Sultankulova KT4, Kopeyev SK5, Su YCF6, Smith GJD7,8,9, Orynbayev MB10.

Author information: 1 Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169857, Singapore. ian.mendenhall@duke-nus.edu.sg. 2 Research Institute for Biological Safety Problems, 080409 Gvardeiskiy, Kordaiskiy rayon, Zhambylskaya oblast, Kazakhstan. aslan_kerim@mail.ru. 3 Research Institute for Biological Safety Problems, 080409 Gvardeiskiy, Kordaiskiy rayon, Zhambylskaya oblast, Kazakhstan. vstrochkov@gmail.com. 4 Research Institute for Biological Safety Problems, 080409 Gvardeiskiy, Kordaiskiy rayon, Zhambylskaya oblast, Kazakhstan. sultankul70@mail.ru. 5 Research Institute for Biological Safety Problems, 080409 Gvardeiskiy, Kordaiskiy rayon, Zhambylskaya oblast, Kazakhstan. kopeyev85@mail.ru. 6 Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169857, Singapore. yvonne.su@duke-nus.edu.sg. 7 Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169857, Singapore. gavin.smith@duke-nus.edu.sg. 8 SingHealth Duke-NUS Global Health Institute, SingHealth Duke-NUS Academic Medical Centre, Singapore 168753, Singapore. gavin.smith@duke-nus.edu.sg. 9 Duke Global Health Institute, Duke University, Durham, NC 27710, USA. gavin.smith@duke-nus.edu.sg. 10 Research Institute for Biological Safety Problems, 080409 Gvardeiskiy, Kordaiskiy rayon, Zhambylskaya oblast, Kazakhstan. omb65@mail.ru.

 

Abstract

Coronaviruses are positive-stranded RNA viruses that infect a variety of hosts, resulting in a range of symptoms from gastrointestinal illness to respiratory distress. Bats are reservoirs for a high diversity of coronaviruses, and focused surveillance detected several strains genetically similar to MERS-coronavirus, SARS-coronavirus, and the human coronaviruses 229E and NL63. The bat fauna of central Asia, which link China to eastern Europe, are relatively less studied than other regions of the world. Kazakhstan is the world’s ninth largest country; however, little is understood about the prevalence and diversity of bat-borne viruses. In this study, bat guano was collected from bat caves in three different sites of southern Kazakhstan that tested positive for coronaviruses. Our phylogenetic reconstruction indicates these are novel bat coronaviruses that belong to the genus Alphacoronavirus. In addition, two distinct lineages of Kazakhstan bat coronaviruses were detected. Both lineages are closely related to bat coronaviruses from China, France, Spain, and South Africa, suggesting that co-circulation of coronaviruses is common in multiple bat species with overlapping geographical distributions. Our study highlights the need for collaborative efforts in understudied countries to increase integrated surveillance capabilities toward better monitoring and detection of infectious diseases.

KEYWORDS: Hypsugo savii; Kazakhstan; Myotis blythii; coronavirus; phylogeny; virus discovery

PMID: 30999711 DOI: 10.3390/v11040356

Keywords: Coronavirus; Alphacoronavirus; Bats; Kazakhstan.

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