Hamster and ferret experimental infection with intranasal low dose of a single strain of SARS-CoV-2
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
Understanding the pathogenesis of the SARS-CoV-2 infection is key to developing preventive and therapeutic strategies against COVID-19, in the case of severe illness but also when the disease is mild. The use of appropriate experimental animal models remains central in the in vivo exploration of the physiopathology of infection and antiviral strategies. This study describes SARS-CoV-2 intranasal infection in ferrets and hamsters with low doses of low-passage SARS-CoV-2 clinical French isolate UCN19, describing infection levels, excretion, immune responses and pathological patterns in both animal species. Individual infection with 10 3 p.f.u. SARS-CoV-2 induced a more severe disease in hamsters than in ferrets. Viral RNA was detected in the lungs of hamsters but not of ferrets and in the brain (olfactory bulb and/or medulla oblongata) of both species. Overall, the clinical disease remained mild, with serological responses detected from 7 days and 10 days post-inoculation in hamsters and ferrets respectively. The virus became undetectable and pathology resolved within 14 days. The kinetics and levels of infection can be used in ferrets and hamsters as experimental models for understanding the pathogenicity of SARS-CoV-2, and testing the protective effect of drugs.
Article activity feed
-
SciScore for 10.1101/2020.09.24.311977: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: These experiments were approved by the Anses/ENVA/UPEC ethic committee and the French Ministry of Research (Apafis n°24818-2020032710416319). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable (Mustela putorius furo, ten neutered males and five females Euroferrets, Denmark) and twenty-one 8-week old female hamsters (Mesocricetus auratus, strain RjHan:AURA - Janvier Labs, France) were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Specific antibody binding was detected by peroxydase-labelled Goat anti-Hamster IgG (H+L) (Invitrogen) and peroxydase-labelled Goat … SciScore for 10.1101/2020.09.24.311977: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: These experiments were approved by the Anses/ENVA/UPEC ethic committee and the French Ministry of Research (Apafis n°24818-2020032710416319). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable (Mustela putorius furo, ten neutered males and five females Euroferrets, Denmark) and twenty-one 8-week old female hamsters (Mesocricetus auratus, strain RjHan:AURA - Janvier Labs, France) were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Specific antibody binding was detected by peroxydase-labelled Goat anti-Hamster IgG (H+L) (Invitrogen) and peroxydase-labelled Goat anti-Ferret IgG (H+L) (KPL), diluted 1:5000 and 1:100 respectively. anti-Hamster IgGsuggested: Noneanti-Ferret IgGsuggested: (Rockland Cat# 618-100-012, RRID:AB_218731)Experimental Models: Cell Lines Sentences Resources Vero CCL-81 cells (passage 32, from ATCC, USA), grown at 80% confluence level were inoculated with 200µl micro-filtered elution. CCL-81suggested: NoneCell supernatants (12 ml) were harvested at day 3 after inoculation and immediately used for passage 1 (P1) produced in T75 culture flasks containing Vero cells as previously described. Verosuggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)Virus titration: Viral load was determined for viral inoculum and for a subset of samples (list in the supplementary table) by plaque assay on VeroE6 cells. VeroE6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Software and Algorithms Sentences Resources The efficiency, slope and correlation coefficient (R2) were determined with the Rotor Gene software. Rotor Genesuggested: None: Wilcoxon signed-rank test was performed by package stat in R (version 3.3.3) to compare viral RNA quantities in nasal washes between males and females ferrets and with GraphPad Prism v6 to compare the areas under the curves for the kinetics of viral presence in clinical samples and of weight changes from D0. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
-
-
SciScore for 10.1101/2020.09.24.311977: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement These experiments were approved by the Anses/ENVA/UPEC ethic committee and the French Ministry of Research (Apafis n°24818-2020032710416319). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable (Mustela putorius furo, ten neutered males and five females Euroferrets, Denmark) and twenty-one 8-week old female hamsters (Mesocricetus auratus, strain RjHan:AURA - Janvier Labs, France) were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Specific antibody binding was detected by peroxydase-labelled Goat anti-Hamster IgG (H+L) (Invitrogen) and peroxydase-labelled Goat anti-Ferret … SciScore for 10.1101/2020.09.24.311977: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement These experiments were approved by the Anses/ENVA/UPEC ethic committee and the French Ministry of Research (Apafis n°24818-2020032710416319). Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable (Mustela putorius furo, ten neutered males and five females Euroferrets, Denmark) and twenty-one 8-week old female hamsters (Mesocricetus auratus, strain RjHan:AURA - Janvier Labs, France) were used. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Specific antibody binding was detected by peroxydase-labelled Goat anti-Hamster IgG (H+L) (Invitrogen) and peroxydase-labelled Goat anti-Ferret IgG (H+L) (KPL), diluted 1:5000 and 1:100 respectively. anti-Hamster IgGsuggested: Noneanti-Ferret IgGsuggested: (Rockland Cat# 618-100-012, RRID:AB_218731)KPLsuggested: NoneExperimental Models: Cell Lines Sentences Resources Vero CCL-81 cells (passage 32, from ATCC, USA), grown at 80% confluence level were inoculated with 200µl micro-filtered elution. CCL-81suggested: NoneCell supernatants (12 ml) were harvested at day 3 after inoculation and immediately used for passage 1 (P1) produced in T75 culture flasks containing Vero cells as previously described. Verosuggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)Non-infected Vero E6 lysate was also coated to provide sample background OD values. Vero E6suggested: RRID:CVCL_XD71)Software and Algorithms Sentences Resources The efficiency, slope and correlation coefficient (R2) were determined with the Rotor Gene software. Rotor Genesuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.
-
