SARS-CoV-2 infection, disease and transmission in domestic cats
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Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the cause of Coronavirus Disease 2019 (COVID-19) and responsible for the current pandemic. Recent SARS-CoV-2 susceptibility and transmission studies in cats show that the virus can replicate in these companion animals and transmit to other cats. Here, we present an in-depth study of SARS-CoV-2 infection, associated disease and transmission dynamics in domestic cats. Six 4- to 5-month-old cats were challenged with SARS-CoV-2 via intranasal and oral routes simultaneously. One day post challenge (DPC), two sentinel contact cats were co-mingled with the principal infected animals. Animals were monitored for clinical signs, clinicopathological abnormalities and viral shedding throughout the 21 DPC observation period. Postmortem examinations were performed at 4, 7 and 21 DPC to investigate disease progression. Viral RNA was not detected in blood but transiently in nasal, oropharyngeal and rectal swabs and bronchoalveolar lavage fluid as well as various tissues. Tracheobronchoadenitis of submucosal glands with the presence of viral RNA and antigen was observed in airways of the infected cats on 4 and 7 DPC. Serology showed that both, principal and sentinel cats, developed SARS-CoV-2-specific and neutralizing antibodies to SARS-CoV-2 detectable at 7 DPC or 10 DPC, respectively. All animals were clinically asymptomatic during the course of the study and capable of transmitting SARS-CoV-2 to sentinels within 2 days of comingling. The results of this study are critical for our understanding of the clinical course of SARS-CoV-2 in a naturally susceptible host species, and for risk assessment of the maintenance of SARS-CoV-2 in felines and transmission to other animals and humans.
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SciScore for 10.1101/2020.08.04.235002: (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 Several veterinary pathologists independently examined slides and were blinded to the treatment groups. 2.9. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The neutralizing antibody titer was recorded as the highest serum dilution at which at least 50% of wells showed virus neutralization (NT50) based on the appearance of CPE observed under a microscope at 72 h post infection. 2.7. Detection of SARS-CoV-2 antibodies by indirect ELISA: To detect SARS-CoV-2 antibodies in sera, indirect ELISAs were performed with the … SciScore for 10.1101/2020.08.04.235002: (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 Several veterinary pathologists independently examined slides and were blinded to the treatment groups. 2.9. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The neutralizing antibody titer was recorded as the highest serum dilution at which at least 50% of wells showed virus neutralization (NT50) based on the appearance of CPE observed under a microscope at 72 h post infection. 2.7. Detection of SARS-CoV-2 antibodies by indirect ELISA: To detect SARS-CoV-2 antibodies in sera, indirect ELISAs were performed with the recombinant viral proteins, nucleoprotein (N) and the receptor-binding domain (RBD), which were produced in-house. NT50suggested: NoneThe wells were washed 3 times with PBS-T, then 100 µl of Goat anti-Feline IgG (H+L) Secondary Antibody, HRP (ThermoFisher Scientific, catalogue number A18757, Waltham, MA, USA) diluted 1:2500 was added to each well and incubated for 1 h at room temperature. anti-Feline IgGsuggested: (Thermo Fisher Scientific Cat# A18757, RRID:AB_2535534)SARS-CoV-2-specific immunohistochemistry (IHC): For IHC, four-micron sections of formalin-fixed paraffin-embedded tissue were mounted on positively charged Superfrost® Plus slides and subjected to IHC using a SARS-CoV-2-specific anti-nucleocapsid mouse monoclonal antibody (clone 6F10, BioVision, Inc. anti-nucleocapsidsuggested: NoneSections were then incubated with the primary antibody (diluted at 1 µg/ml in Antibody Diluent [Dako, Carpinteria, CA]) for 30 min at room temperature, followed by a polymer-labeled goat anti-mouse IgG coupled with alkaline phosphatase (30 minutes; Powervision anti-mouse IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources The 200 µl per well of virus sera mixture was then cultured on VeroE6 cells in 96-well plates. VeroE6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Sections from mock- and SARS-CoV-2-infected Vero cell pellets were used as negative and positive assay controls. Verosuggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)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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