Susceptibility of sheep to experimental co-infection with the ancestral lineage of SARS-CoV-2 and its alpha variant
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SciScore for 10.1101/2021.11.15.468720: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Ethics statement: All animal studies and experiments were approved and performed under the Kansas State University (KSU) Institutional Biosafety Committee (IBC, Protocol #1460) and the Institutional Animal Care and Use Committee (IACUC, Protocol #4508.2) in compliance with the Animal Welfare Act. Sex as a biological variable Virus challenge of animals: Ten male sheep, approximately 6 months of age, were acquired from Frisco Farms (Ewing, IL) and acclimated for ten days in BSL-3Ag biocontainment with feed and water ad libitum prior to experimental procedures. Randomization not detected. Blinding Two independent veterinary pathologists (blinded to the treatment groups) examined the slides and … SciScore for 10.1101/2021.11.15.468720: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Ethics statement: All animal studies and experiments were approved and performed under the Kansas State University (KSU) Institutional Biosafety Committee (IBC, Protocol #1460) and the Institutional Animal Care and Use Committee (IACUC, Protocol #4508.2) in compliance with the Animal Welfare Act. Sex as a biological variable Virus challenge of animals: Ten male sheep, approximately 6 months of age, were acquired from Frisco Farms (Ewing, IL) and acclimated for ten days in BSL-3Ag biocontainment with feed and water ad libitum prior to experimental procedures. Randomization not detected. Blinding Two independent veterinary pathologists (blinded to the treatment groups) examined the slides and morphological descriptions were provided. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Following three washes with PBS-T, 100 μL of HRP-labelled Rabbit Anti-Sheep IgG (H+L) secondary antibody (VWR, Batavia, IL, USA) diluted 1:1000 (100ng/mL) was added to each well and incubated for 1 hour at RT. Anti-Sheep IgGsuggested: NoneIndirect ELISA was used to detect bovine coronavirus (BCoV) antibodies in sera with Spike (S) recombinant viral protein (LSBio, Seattle, WA, USA) using the methods described above. BCoVsuggested: NoneSARS-CoV-2-specific immunohistochemistry (IHC): IHC was performed as previously described (29) on four-micron sections of formalin-fixed paraffin-embedded tissue mounted on positively charged Superfrost® Plus slides and subjected to IHC using a SARS-CoV-2-specific anti-nucleocapsid rabbit polyclonal antibody (3A, developed by our laboratory) with the method previously described (30). anti-nucleocapsidsuggested: NoneExperimental Models: Cell Lines Sentences Resources Virus isolation was performed by culturing 100 µl of filtered (0.2 µm; MidSci, St. Louis, MO) sample /well in duplicate on Vero E6/TMPRRS2 cells and monitoring for CPE for up to 5 days post inoculation. Vero E6/TMPRRS2suggested: NoneSusceptibility of ovine and bovine cells to SARS-CoV-2: The SARS-CoV-2 USA-WA1/2020 strain was passaged 3 times in Vero-E6 cells to establish a stock virus for infection experiments. Vero-E6suggested: NoneCell supernatants were titrated on Vero E6 cells to determine SARS-CoV-2 replication kinetics by virus titers (TCID50/mL). Vero E6suggested: NoneThe mixture was subsequently cultured on Vero-E6/TMPRSS2 cells in 96-well plates. Vero-E6/TMPRSS2suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Software and Algorithms Sentences Resources Swabs were placed in 2mL of viral transport medium (DMEM, Corning; combined with 1% antibiotic-antimycotic, ThermoFisher), vortexed, and aliquoted directly into cryovials and RNA stabilization/lysis Buffer RLT (Qiagen, Germantown, MD, USA). ThermoFishersuggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)To determine an accurate relative percentage of each SARS-CoV-2 lineage in each sample, BLAST databases were first generated from individual trimmed and filtered sample reads. BLASTsuggested: (BLASTX, RRID:SCR_001653), McKinney, TX, for 4-7 days at room temperature), trachea, and lungs as well as various other extrapulmonary tissues (liver, spleen, kidneys, heart, pancreas, gastrointestinal tract [stomach, small intestine including Peyer’s patches and colon], cerebrum [including olfactory bulb], tonsils and numerous lymph nodes) were routinely processed and embedded in paraffin. Peyer’ssuggested: 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.
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.
Results from scite Reference Check: We found no unreliable references.
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