SARS-CoV-2 surveillance in Norway rats ( Rattus norvegicus ) from Antwerp sewer system, Belgium

  1. Valeria Carolina Colombo
  2. Vincent Sluydts
  3. Joachim Mariën
  4. Bram Vanden Broecke
  5. Natalie Van Houtte
  6. Wannes Leirs
  7. Lotte Jacobs
  8. Arne Iserbyt
  9. Marine Hubert
  10. Leo Heyndrickx
  11. Hanne Goris
  12. Peter Delputte
  13. Naomi De Roeck
  14. Joris Elst
  15. Robbert Boudewijns
  16. Kevin K. Ariën
  17. Herwig Leirs
  18. Sophie Gryseels

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Abstract

Background

SARS-CoV-2 human-to-animal transmission can lead to the establishment of novel reservoirs and the evolution of new variants with the potential to start new outbreaks in humans.

Aim

We tested Norway rats inhabiting the sewer system of Antwerp, Belgium, for the presence of SARS-CoV-2 following a local COVID-19 epidemic peak. In addition, we discuss the use and interpretation of SARS-CoV-2 serological tests on non-human samples.

Methods

Between November and December 2020, Norway rat oral swabs, feces and tissues from the sewer system of Antwerp were collected to be tested by RT-qPCR for the presence of SARS-CoV-2. Serum samples were screened for the presence of anti-SARS-CoV-2 IgG antibodies using a Luminex microsphere immunoassay (MIA). Samples considered positive were then checked for neutralizing antibodies using a conventional viral neutralization test (cVNT).

Results

The serum of 35 rats was tested by MIA showing 3 potentially positive sera that were later shown to be negative by cVNT. All tissue samples of 39 rats analyzed tested negative for SARS-CoV-2 RNA.

Conclusion

This is the first study that evaluates SARS-CoV-2 infection in urban rats. We can conclude that the sample of 39 rats had never been infected with SARS-CoV-2. We show that diagnostic serology tests can give misleading results when applied on non-human samples. SARS-CoV-2 monitoring activities should continue due to the emergence of new variants prone to infect Muridae rodents.

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  1. ScreenIT

    SciScore for 10.1101/2021.03.06.433708: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: All procedures were carried out under the approval of the University of Antwerp Ethical Committee for Animal Experiments (ECD code 2020-21).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Serology: To test SARS-CoV-2 exposure in sewer rats, serum samples were first screened for the presence of binding anti-SARS-CoV-2 IgG antibodies, using an in-house Luminex microsphere immunoassay (MIA) (Mariën et al 2021).
    anti-SARS-CoV-2 IgG
    suggested: None
    MIA
    suggested: None
    A biotin-labelled goat anti-mouse IgG Y-chain specific conjugate (Sigma, B7022, 1/300 dilution) was used for visualization of the primary antibodies.
    A biotin-labelled goat anti-mouse IgG Y-chain specific conjugate
    suggested: None
    anti-mouse IgG
    suggested: (Sigma-Aldrich Cat# B7022, RRID:AB_258598)
    B7022
    suggested: (Assay Biotech Cat# B7022, RRID:AB_10684184)

    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2021.03.06.433708 on bioRxiv