Association between mink coronavirus (MCoV), Campylobacter spp., and diarrhea in farmed mink (Neogale vison)

Michelle Lauge Quaade; Mikael Leijon; Mikhayil Hakhverdyan; Thomas Bruun Rasmussen; Charlotte Kristiane Hjulsager; Lars Andresen; Karin Mundbjerg; Anne Sofie Vedsted Hammer

doi:10.1371/journal.pone.0341271

Peer Review History

Original SubmissionJuly 21, 2025
4 Nov 2025 Decision Letter - Baochuan Lin, Editor Dear Dr. Quaade, Please submit your revised manuscript by Dec 19 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. 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If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: The manuscript under review meets all the requirements necessary for publication in the journal. It has been prepared with great care, in accordance with all the rigors of scientific writing. Its advantages include not only a summary of the results of zoonotic safety research conducted to date on Danish American mink farms, but also an indication of further necessary stages of experimental activities in the breeding of the most important species of carnivorous fur animals. Reviewer #2: This manuscript appears to be a good reflection of sound science to examine disease, post-weaning anorexia diarrhea syndrome (PADS) in farmed mink. There are a few areas that need to be clarified: Methods 1.3 Whole Genome Shotgun sequencing is not exactly clear. The procedure is specific for RNA- RNA viruses. In Results section 2.4 this is called Metagenomic shotgun sequencing. Neither description is exactly correct. It is more correctly metagenomic since the filtering of cells and bacteria appears to be a process that was not successful. However, it is also only an examination of isolated RNA sequences. Metagenomes would include the isolation and sequencing of DNA also. Just define this so that readers know you only examined RNA- to find MCoV. I thought there might be information about the Campylobacters based on the metagenomic statement. Related to Campylobacter, there are many species that may reside naturally in the mink. Others have noted that wild animals may harbor specific types of C. jejuni that differ from those associated with domestic animals. Additionally, there may be other Campylobacter species that are specifically found just in the PADS mink. I believe you should note the limitations of only performing qPCR for 16S rRNA detection. ****** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: No Reviewer #2: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation. NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. https://doi.org/10.1371/journal.pone.0341271.r001
Revision 1
17 Dec 2025 Author Response Response to the Academic Editor 1. PLOS ONE Style Requirements Please ensure that your manuscript meets PLOS ONE's style requirements... Response: We have carefully reviewed the PLOS ONE style requirements. We confirm that the manuscript, including file naming conventions and formatting, has been updated to fully adhere to these guidelines. 2. Data Availability Please confirm at this time whether or not your submission contains all raw data required to replicate the results of your study... Response: We have revised the Data Availability Statement to ensure full transparency. The "minimal data set" required to replicate all study findings, including necessary metadata, has been deposited in a stable, public repository. • Revised Data Availability Statement: Data from this study is available at the Open Science Framework (OSF) under study ID m4ce2 and can be accessed via DOI: 10.17605/OSF.IO/M4CE2. All MCoV sequence data has been deposited at NCBI with accessions: PQ451274.1-PQ451290.1. 3. Additional Citations If the reviewer comments include a recommendation to cite specific previously published works... Response: The reviewers did not recommend specific citations in their initial comments. However, to address Reviewer #2’s comment regarding Campylobacter diversity, we have proactively added a relevant reference (Petersen et al., 2001) to support our discussion on wildlife-specific strains. 4. Reference List Please review your reference list to ensure that it is complete and correct. Response: We have thoroughly reviewed the reference list and confirmed it is complete, correct, and compliant with journal formatting. Response to Reviewer #2 Comment 1: Sequencing Terminology Methods 1.3 Whole Genome Shotgun sequencing is not exactly clear... In Results section 2.4 this is called Metagenomic shotgun sequencing. Neither description is exactly correct... Just define this so that readers know you only examined RNA. Response: We agree with the reviewer that the previous terminology was imprecise given that our protocol specifically targeted RNA. To improve clarity and accuracy we have renamed the headers for sections 1.3 and 2.4 to "Next-Generation Sequencing." Comment 2: Campylobacter Specificity and Wildlife Strains Related to Campylobacter, there are many species that may reside naturally in the mink. Others have noted that wild animals may harbor specific types of C. jejuni that differ from those associated with domestic animals... I believe you should note the limitations of only performing qPCR for 16S rRNA detection. Response: We thank the reviewer for this important observation regarding Campylobacter diversity. We agree that a significant limitation of our study is the reliance on a 16S rRNA-based qPCR, which detects the genus but does not distinguish between species. We acknowledge that mink likely harbor natural/commensal Campylobacter species and that specific types—potentially different from those in other domestic animals—might be the true drivers of PADS. We have updated the Discussion (Page 25, Lines 460-470) section to explicitly address this limitation, noting that we could not differentiate between a specific pathogenic species unique to PADS and a general overgrowth of commensal flora. Reviewers comments: Reviewer #2: This manuscript appears to be a good reflection of sound science to examine disease, post-weaning anorexia diarrhea syndrome (PADS) in farmed mink. There are a few areas that need to be clarified: Methods 1.3 Whole Genome Shotgun sequencing is not exactly clear. The procedure is specific for RNA- RNA viruses. In Results section 2.4 this is called Metagenomic shotgun sequencing. Neither description is exactly correct. It is more correctly metagenomic since the filtering of cells and bacteria appears to be a process that was not successful. However, it is also only an examination of isolated RNA sequences. Metagenomes would include the isolation and sequencing of DNA also. Just define this so that readers know you only examined RNA- to find MCoV. I thought there might be information about the Campylobacters based on the metagenomic statement. Response: The titles of paragraphs 1.3 and 2.4 has been changed to “Next-Generation Sequencing” to improve clarity. Related to Campylobacter, there are many species that may reside naturally in the mink. Others have noted that wild animals may harbor specific types of C. jejuni that differ from those associated with domestic animals. Additionally, there may be other Campylobacter species that are specifically found just in the PADS mink. I believe you should note the limitations of only performing qPCR for 16S rRNA detection. Response: We thank the reviewer for this insightful observation regarding the specificity of our assay and the diversity of Campylobacter in wildlife. We fully agree that a significant limitation of our study is the reliance on a 16S rRNA-based qPCR, which detects the genus as a whole but does not distinguish between specific species or strains. We acknowledge that mink likely harbor natural, commensal Campylobacter populations, and as the reviewer notes, studies indicate that wildlife often carry specific strains or clonal lines that differ from those found in humans or other domestic animals. To address this, we have revised the Discussion section (Page 25, Lines 460-470). We now explicitly state that our data cannot distinguish between specific pathogenic types and commensal flora. We have also incorporated a reference to Petersen et al. (2001) to highlight that Campylobacter strains in Danish wildlife have been shown to possess distinct serotype distributions and clonal lines compared to agricultural animals, suggesting that the "wildlife reservoir" may be distinct from the pathogenic strains usually screened for. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0341271.r002
5 Jan 2026 Decision Letter - Baochuan Lin, Editor Association between mink coronavirus (MCoV), Campylobacter spp., and diarrhea in farmed mink (Neogale vison) PONE-D-25-37216R1 Dear Dr. Quaade, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support . If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Baochuan Lin, Ph.D. Academic Editor PLOS One Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #2: N/A ****** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #2: Yes ****** Reviewer #2: The authors have written a nice paper and have addressed the issues that I identified in the original submission. ****** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #2: No ******** https://doi.org/10.1371/journal.pone.0341271.r003
Formally Accepted
Acceptance Letter - Baochuan Lin, Editor PONE-D-25-37216R1 PLOS One Dear Dr. Quaade, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS One. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Baochuan Lin Academic Editor PLOS One https://doi.org/10.1371/journal.pone.0341271.r004

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