Severe acute respiratory disease in American mink ( Neovison vison ) experimentally infected with SARS-CoV-2

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   ScreenIT

   Jan 28, 2022

   SciScore for 10.1101/2022.01.20.477164: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Ethics statement: All animal experiments were approved by the Institutional Animal Care and Use Committee of Rocky Mountain Laboratories, NIH and carried out in an Association for Assessment and Accreditation of Laboratory Animal Care (AALAC
   Sex as a biological variable	The animals used in the infection study consisted of 9 females and two males; intake female body weight range 1.04 kg – 1.47 kg, mean = 1.18 kg, male weights were 2.06 kg and 2.73.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	Contamination: Mycoplasma testing was performed at regular intervals and no mycoplasma was detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   U…

   SciScore for 10.1101/2022.01.20.477164: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Ethics statement: All animal experiments were approved by the Institutional Animal Care and Use Committee of Rocky Mountain Laboratories, NIH and carried out in an Association for Assessment and Accreditation of Laboratory Animal Care (AALAC
   Sex as a biological variable	The animals used in the infection study consisted of 9 females and two males; intake female body weight range 1.04 kg – 1.47 kg, mean = 1.18 kg, male weights were 2.06 kg and 2.73.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	Contamination: Mycoplasma testing was performed at regular intervals and no mycoplasma was detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Upon arrival whole blood from all mink were screened for antibodies against SARS-CoV-2.	SARS-CoV-2 suggested: None
   The secondary antibody was the Vector Laboratories ImPress VR anti-rabbit IgG polymer (cat# MP-6401).	anti-rabbit IgG suggested: None
   To detect ACE-2, ACE-2 Antibody R&D Systems (catalog #AF933) was used at a 1:100 dilution with Vector Laboratories ImPress anti-goat IgG polymer (Cat #MP-7405) as a secondary antibody.	anti-goat IgG suggested: (Vector Laboratories Cat# MP-7405, RRID:AB_2336526)
   Experimental Models: Cell Lines
   Sentences	Resources
   Briefly, plates pre-coated with poly-L-lysine (Sigma-Aldrich, St. Louis, MO, USA) were seeded with 293T cells and transfected the following day with 1,200 ng of empty plasmid and 400 ng of plasmid encoding coronavirus spike or no-spike plasmid control (green fluorescent protein (GFP)).	293T suggested: None
   BHK cells were seeded in black 96-well plates and transfected the next day with 100 ng plasmid DNA encoding human or mink ACE2, using polyethylenimine (Polysciences, Inc., Warrington, PA, USA).	BHK suggested: None
   After a 1-hour incubation at 37°C and 5% CO2, the virus-serum mixture was added to VeroE6 cells.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Recombinant DNA
   Sentences	Resources
   These spike sequences were synthesized and cloned into pcDNA3.1+ (GenScript Biotech, Piscataway, NJ, USA)	pcDNA3.1+ suggested: RRID:Addgene_117272)
   BHK cells previously transfected with ACE2 plasmid of interest were inoculated with equivalent volumes of pseudotype stocks.	ACE2 suggested: RRID:Addgene_164219)
   Software and Algorithms
   Sentences	Resources
   Reads were de novo assembled using SPAdes v.	SPAdes suggested: (SPAdes, RRID:SCR_000131)
   The percent identity between the ACE2 sequences was calculated by Clustal Omega (39)	Clustal Omega suggested: (Clustal Omega, RRID:SCR_001591)
   Mutagenesis to show residues that differ in mink ACE2, and the alpha and delta variant RBD, was performed in COOT (40).	COOT suggested: (Coot, RRID:SCR_014222)
   The figures were generated using The Pymol Molecular Graphics System (https://www.schrodinger.com/pymol).	Pymol suggested: (PyMOL, RRID:SCR_000305)
   ARTIC primers and Illumina adapters were trimmed, low quality bases and duplicate reads were filtered out, and mapping and variant calling were completed as described in the iVar and PrimalSeq pipeline described by Grubaugh et al.(47).	PrimalSeq suggested: None
   Statistical analysis: Statistical analysis was performed using GraphPad Version 8.4.3.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)

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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: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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

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   Version published to 10.1101/2022.01.20.477164v1 on bioRxiv

   Jan 24, 2022