SARS-CoV-2 infection and transmission in the North American deer mouse
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Abstract
Widespread circulation of SARS-CoV-2 in humans raises the theoretical risk of reverse zoonosis events with wildlife, reintroductions of SARS-CoV-2 into permissive nondomesticated animals. Here we report that North American deer mice ( Peromyscus maniculatus ) are susceptible to SARS-CoV-2 infection following intranasal exposure to a human isolate, resulting in viral replication in the upper and lower respiratory tract with little or no signs of disease. Further, shed infectious virus is detectable in nasal washes, oropharyngeal and rectal swabs, and viral RNA is detectable in feces and occasionally urine. We further show that deer mice are capable of transmitting SARS-CoV-2 to naïve deer mice through direct contact. The extent to which these observations may translate to wild deer mouse populations remains unclear, and the risk of reverse zoonosis and/or the potential for the establishment of Peromyscus rodents as a North American reservoir for SARS-CoV-2 remains unknown.
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SciScore for 10.1101/2020.07.25.221291: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Experiment were approved by the Animal Care Committee located at the Canadian Science Center for Human and Animal Health in accordance with the guidelines provided by the Canadian Council on Animal Care. Randomization Deer mice were randomly assigned to their respective groups and were housed in a temperature-controlled, light-cycled facility. Blinding not detected. Power Analysis not detected. Sex as a biological variable Thirteen to thirty two-week old male or female deer mice were infected with 105 or 106 TCID50 of SARS-CoV-2 by an intranasal route (i.n.) of administration in a 50 μl volume. Cell Line Authentication Contamination: The P1 virus was subsequently … SciScore for 10.1101/2020.07.25.221291: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Experiment were approved by the Animal Care Committee located at the Canadian Science Center for Human and Animal Health in accordance with the guidelines provided by the Canadian Council on Animal Care. Randomization Deer mice were randomly assigned to their respective groups and were housed in a temperature-controlled, light-cycled facility. Blinding not detected. Power Analysis not detected. Sex as a biological variable Thirteen to thirty two-week old male or female deer mice were infected with 105 or 106 TCID50 of SARS-CoV-2 by an intranasal route (i.n.) of administration in a 50 μl volume. Cell Line Authentication Contamination: The P1 virus was subsequently passaged at a 1:1000 dilution on mycoplasma-free VeroE6 cells (ATCC) in Dulbecco’s Modified Eagle’s Medium (Hyclone) containing 1% L-glutamine and 0.5 μg/ml of TPCK-trypsin and harvested when 80% cytopathic effect (CPE) became evident. Table 2: Resources
Antibodies Sentences Resources SARS-CoV-2-S-specific enzyme-linked immunosorbent assay (ELISA): SARS-CoV-2 spike/nucleoprotein (S/N)-specific IgG antibody responses were assessed using an in-house assay. S/N)-specific IgGsuggested: NoneDeer mouse IgG was detected with a KPL peroxidase-labeled polyclonal goat antibodies against Peromyscus leucopus IgG (H+L) (Sera Care) antibodies against Peromyscus leucopus IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources The virus stock was titrated on Vero cells by conventional TCID50 assay, as described previously53. Verosuggested: NoneThe sera-virus mixtures were added to 24-well plates containing Vero E6 cells at 100% confluence, followed by incubation at 37 °C and 5% CO2 for 1 hour. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources GraphPad Prism’s multiple t test was used to perform the second subtraction so as not to lose the variation of the mock animals. GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)For the phylogenetic analysis, mitochondrial genome sequences (see Accession codes) were aligned with MAFFT v7.46759, with regions of poor alignment trimmed with Gblocks v0.91b60 resulting in a final alignment of 15,393bp in 115 blocks of minimum 5bp lengths. MAFFTsuggested: (MAFFT, RRID:SCR_011811)Gblockssuggested: (Gblocks, RRID:SCR_015945)A maximum likelihood phylogeny was constructed with RAxML v8.2.1261 using the GTR+I+G4 substitution model as selected by modeltest-ng62. RAxMLsuggested: (RAxML, RRID:SCR_006086)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.
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SciScore for 10.1101/2020.07.25.221291: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Here, eight to thirty two-week old male or female deer mice (P. maniculatus rufinus; in-house colony) were inoculated with 105 TCID50 of SARS-CoV-2 by an intranasal route (i.n.) and monitored daily for clinical signs and weight loss for 21 days or necropsied at 2 and 4 day postinfection (dpi). Table 2: Resources
Antibodies Sentences Resources All five deer mice infected at 105 TCID50 had detectable serum IgG titers against mixed spike/nucleoprotein (S/N) antigen as assessed by ELISA at 14 dpi (OD 2.7 – 2.8 at 1:100) (Fig. 4g), and … SciScore for 10.1101/2020.07.25.221291: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Here, eight to thirty two-week old male or female deer mice (P. maniculatus rufinus; in-house colony) were inoculated with 105 TCID50 of SARS-CoV-2 by an intranasal route (i.n.) and monitored daily for clinical signs and weight loss for 21 days or necropsied at 2 and 4 day postinfection (dpi). Table 2: Resources
Antibodies Sentences Resources All five deer mice infected at 105 TCID50 had detectable serum IgG titers against mixed spike/nucleoprotein (S/N) antigen as assessed by ELISA at 14 dpi (OD 2.7 – 2.8 at 1:100) (Fig. 4g), and neutralizing antibodies by 28 dpi (plaque reduction neutralization test (PRNT90) 1:40 to 1:320) (Fig. 4h). PRNT90suggested: NoneData from additional tools added to each annotation on a weekly basis.
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