Exposure route, sex, and age influence disease outcome in a golden Syrian hamster model of SARS-CoV-2 infection

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   ScreenIT

   Jun 16, 2021

   SciScore for 10.1101/2021.06.12.448196: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics	IACUC: All experiments performed under Animal User Document H-20-006, approved by the Animal Care Committee at the Canadian Science Center for Human and Animal Health in accordance with the guidelines provided by the Canadian Council on Animal Care. Euthanasia Agents: All procedures were performed under inhalation anesthesia using isoflurane.
   Sex as a biological variable	Hamster challenge and re-challenge experiments: Groups of male or female golden Syrian hamsters were anaesthetized and exposed to 1×105 TCID50 SARS-CoV-2 by an intranasal route of inoculation with the inoculum distributed evenly into both nares in a volume of 20 µl (intranasal low volume) or in a volume of 100 µl to allow for …

   SciScore for 10.1101/2021.06.12.448196: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics	IACUC: All experiments performed under Animal User Document H-20-006, approved by the Animal Care Committee at the Canadian Science Center for Human and Animal Health in accordance with the guidelines provided by the Canadian Council on Animal Care. Euthanasia Agents: All procedures were performed under inhalation anesthesia using isoflurane.
   Sex as a biological variable	Hamster challenge and re-challenge experiments: Groups of male or female golden Syrian hamsters were anaesthetized and exposed to 1×105 TCID50 SARS-CoV-2 by an intranasal route of inoculation with the inoculum distributed evenly into both nares in a volume of 20 µl (intranasal low volume) or in a volume of 100 µl to allow for inhalation of the inoculum (intranasal high volume), or by an intragastric route of infection via oral gavage.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   Cells and Viruses: Vero cells (ATCC) were cultured in Minimal Essential Medium (MEM) (Hyclone) supplemented with 5% Bovine Growth Serum Supplemented Calf (Hyclone) and L-glutamine.	Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
   SARS-CoV-2 (Canada/ON-VIDO-01/2020; EPI_ISL_425177) was isolated from a positive patient sample and stocks of virus were grown in VeroE6 cells.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Software and Algorithms
   Sentences	Resources
   Data analysis: Results were analyzed and graphed using Prism 8 software (Graphpad Software).	Prism suggested: (PRISM, RRID:SCR_005375) Graphpad suggested: (GraphPad, RRID:SCR_000306)

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   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).

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   A limitation of the routes of administration described in this work is that i.n. adminstration results in some inoculum being swallowed and entering the gastrointestinal tract54. Even with low volume i.n. administration, there may be a small amount of virus that become deposited within the lower respiratory tract shortly after infection. With the i.g. instillation by oral gavage some inoculum is also expected to be introduced to the respiratory tract52 or oral cavity when the feeding tube is inserted or withdrawn. We questioned whether the differential pathogenesis observed with an i.g. route versus an high-volume intranasal route administration was due to the alimentary tract exposure or rather a small viral dose seeding the airway during gavage; however, a recent report describes that low dose SARS-CoV-2 inoculation of the hamster airway results in diminished pathogenesis55. Given that the i.n.L inoculation may also be expected to result in a low dose delivery of virus to the lower respiratory tract, the milder clinical signs of illness that we observed are consistent with the reduced pathogenesis reported by others upon low dose exposure. Given our findings that all routes of infection resulted in similar timing and extent of virus replication in the respiratory and digestive tract tissues (Fig. 1), it is possible that disease outcome is more directly related to the nature of the immune responses that are established at the initial site of greatest virus exposure and that ...

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2021.06.12.448196v1 on bioRxiv

   Jun 16, 2021