-->PONE-D-25-43850-->-->Brucella canis  Outbreak Managment Towards Rehoming Strategies: The Italian Experience-->-->PLOS ONE

Dear Dr. D'Aurelio,

Thank you for submitting your manuscript to PLOS ONE . Two expert reviewers evaluated your paper and their comments are provided below. Both expressed the opinion that the information provided represents an important contribution to the field. But both also pointed out some issues that need to be addressed before the paper will be considered acceptable for publication in the journal. Consequently, I am going to ask that you submit a revised version and adequately and appropriately addresses all of  the comments raised by both reviewers.

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Academic Editor

PLOS ONE

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- doi: 10.12834/VetIt.2561.16874.1

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Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Yes

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-->2. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: N/A

Reviewer #2: N/A

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-->3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

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-->4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: Yes

Reviewer #2: Yes

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-->5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: General Introduction comment: I think the introduction could be tightened up a bit. Right now, there’s quite a lot of detail on the global distribution of B. canis, which could be shortened. What feels more central here is Italy’s unique regulatory context and why that made a no-kill strategy necessary. Section 2.2 (“Implementation of the No-Kill Strategy”) would actually fit better in the introduction, since it sets the stage for why this protocol was needed in the first place.

2.5 Post-Adoption Monitoring and Owner Responsibilities: It would be useful to clarify how compliance with follow-up testing was verified (e.g., submission of veterinary reports)

2.4 Seronegative animals were selected for potential rehoming: The quarantine and sequential testing scheme is clearly described, but the rationale for choosing the specific timepoints (days 10, 28, and 56/8 weeks) should be explained. It would also strengthen the section to indicate how many animals tested positive at each stage and to briefly describe housing and welfare conditions during quarantine.

Results section: In the first paragraph of the Results, you report how many dogs tested positive or negative. I think it would give readers a clearer picture if you also included some basic demographics for those groups, like sex ratios or age distribution.

4.5 Study Limitations: The limitations section is strong, but it would be helpful to expand on how the ~20% loss to follow-up could have biased the results, since dogs lost to monitoring may have differed from the ones that followed up. It would also be helpful to explicitly discussing the impact of imperfect test sensitivity and the potential for false negatives would improve the transparency and balance of the interpretation.

Grammar:

1) Brucella canis Outbreak Management (not managment) Towards Rehoming Strategies

2) Introduction first paragraph: (Djokic et al., 2023;) has a stray ; and should be delated and say (Djokic et al., 2023)

3) In the last paragraph of the introduction, there is an et al that does not appear to go with any sentences. Needs to be deleted.

4) Methods: "seronegative status was determined using mSAT, with a cut-off titer of 1:20" (not cu-toff)

5) 96,7% should be a decimal not a comma (96.7%)

Reviewer #2: Overview

The aim of this article is to is to describe the procedure used to manage a Brucella canis outbreak in dogs in Italy. Due to ethical and legislative reasons, the authors chose a no-kill approach with strict serological and bacterial testing of animals prior to selection of consistently negative dogs for rehoming.

According to the authors, of 683 tested dogs, 254 were positive by mSAT and/or bacteriological culture while 438 were negative by both tests. Using the no-kill strategy 398 negative dogs were rehomed.

Major comments

Lines 40 to 51: Insert the references where appropriate for easier identification of which reference supports which statement.

Table 1: As there is a clear difference in management between males and female dogs having the test results by sex would provide more information to the reader.

Minor comments

Throughout the manuscript: Improper use of grammar and spelling mistakes. Please check.

Conclusion

This work has potential for good impact in the scientific community, and we recommend this paper with minor revisions.

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Reviewer #1: No

Reviewer #2: No

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-->PONE-D-25-43850R1-->-->Brucella canis  Outbreak Management Towards Rehoming Strategies: The Italian Experience-->-->PLOS One

Dear Dr. D'Aurelio,-->--> -->-->Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.-->--> -->-->Both reviewers expressed the opinion that your manuscript was greatly improved and that you had addressed the specific concerns raised in their previous reviews. Both, however, have made suggestions for minor grammatical and typographical revisions aimed at improving the 'polish' of  the final version of  the paper. Considering that PLOS ONE leaves this last step to the authors and not to professional editors, I am going to ask that you submit a revised version of  the manuscript addressing the reviewers' suggestions. Given that these are minor changes they should not take long to address.

Please submit your revised manuscript by Jun 08 2026 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.

Please include the following items when submitting your revised manuscript:-->

• A 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'.

-->

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

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We look forward to receiving your revised manuscript!

Sincerely,

Marty Roop, Ph.D.

Academic Editor

PLOS One

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If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. 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.

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Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.-->

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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-->2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->3. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: No

Reviewer #2: Yes

**********

-->6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: The authors have addressed the reviewer comments thoroughly and substantially improved the manuscript, particularly in the Methods, Results, and Limitations sections. The study is well-structured and provides valuable insight into B. canis outbreak management. A few minor grammatical issues still remain. Pending these minor revisions, I recommend acceptance.

1) "discospondlytis”: should be “discospondylitis

2) Bracket error: “Africa 12], and Europe [1]”

3) Miss space after "programs: "absence of mandatory surveillance programs[1]"

4) This sentence appears twice so need to delete one instance: “Because B. canis is characterized by intermittent shedding…”

5) Broken/fragmented sentence and not sure where the et al was suppose to go to: "In 2020, B. canis has been detected and isolated for the first time in a commercial breeding kennel [18]. et al."

Reviewer #2: Overview

Thank you for your careful revision and detailed response. The manuscript is substantially improved in clarity and methodological transparency. Most of my prior concerns have been addressed; just a few edits are required.

Point-by-point assessment relative to my previous review:

Major comments

Lines 40 to 51: Insert the references where appropriate for easier identification of which reference supports which statement.

- Assessment: The authors included references for each statement.

Table 1: As there is a clear difference in management between males and female dogs having the test results by sex would provide more information to the reader.

- Assessment: The authors have included additional tables which provide a breakdown of the serological and culture results per sex for better understanding.

Minor comments

Line 59: It seems that the sentence starts with a “et al.” which does not seem connected to any sentence. It needs to be removed.

Table 1: The first line of the table reads “Serology (Msat)” where it should read “Serology (mSAT)”. Please make the appropriate change.

**********

-->7. PLOS authors have the option to publish the peer review history of their article (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

**********

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Brucella canis  Outbreak Management Towards Rehoming Strategies: The Italian Experience

PONE-D-25-43850R2

Dear Dr. D'Aurelio,

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.

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Sincerely,

R. Martin Roop II, Ph.D.

Academic Editor

PLOS One

Additional Editor Comments (optional):

Reviewers' comments: