Dear Dr. MBUTHIA,

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Mabel Kamweli Aworh, DVM, MPH, PhD. FCVSN

Academic Editor

PLOS ONE

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In addition to addressing the reviewers comments, please highlight key limitations of the study as the las paragraph of the Discussion section. Also ensure that 80% of the reference list is within the last five years.

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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?

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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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??>

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: General Comment

This paper offers truly important insights into how migratory birds play a role in spreading antimicrobial resistance. Its findings are vital for guiding public health policies and environmental strategies against AMR. What's particularly exciting is that this study uncovers a link between the little stint bird and AMR, thoroughly addressing a big gap in what we currently know. Looking ahead, authors should consider sequencing their isolates, as a future study. It would provide much deeper genomic insights, especially into the mobile genetic elements that carry or share these resistance genes.

Specific Comments

Line 242–243: The table layout appears cluttered, making it hard to read, especially at publication scale. Also, consider defining these abbreviations (for example “C1”, “C2”) mentioned in the caption.

Line 256: Insert a space between “1” and “and.”

Lines 261–262: Similar to Table 1, define all abbreviations for clarity. Improve table formatting for readability.

Lines 274–278: Good you mentioned, potential AMR factors such as human interactions at hot springs, rivers shared with livestock, and salt pans effluent. Consider elaborating on how these factors directly contribute to AMR in migratory birds in the regions. Expand the discussion to explain the mechanisms or pathways through which these factors may directly or indirectly influence AMR transmission or persistence.

Lines 442–443: Review reference formatting (for instance citations, 35 and 39) and ensure consistent use of the appropriate citation style as required by the journal throughout the reference list.

Reviewer #2: The topic of the study is definitely of interest to the AMR community. Minor corrections are suggested as enumerated below

1. Paragraph starting at line 64. Introducing a statement or 2 describing how the little stint (Calidris minuta) specifically feeds and generally interacts with the niche upon arrival at lakes Magadi and Bogoria will be informative.

2. Line 77. "While anthropogenic activities are recognized as a key driver of AMR dissemination by migratory birds globally,", kindly provide reference for this.

3. Line 94. "Anthropogenic activity is substantial due to the commercial trona and soda ash mining industry." Knowing that PLoS One is a generalist publication, the authors might wish to clarify to readers how the commercial trona might contribute to AMR risk at this site.

4. Line 90. "Human interactions with the lake are minimal, with the agro-pastoralists Tugens and animals (domestic and wildlife) accessing the freshwater springs by the lakes." At present, I am not convinced, as per this statement, that the human interaction due to the agro-pastoralists and their animals are minimal (no evidence provided) or minimal compared to lake Magadi. The authors should provide clarifying evidence.

5. Line 117. Reporting the results of number of samples collected under the methods section is inappropriate. Such data should be in the result section.

6. Line 120. We are told that water samples were drawn from lakes, hot springs, rivers, and springs [4 sources] used by the little stint. What we are not told is whether the 48 samples that was eventually collected was achieved using random or other form of sampling from these 4 sources. Please clarify.

7. At line 153, "They included; ampicillin (AMP, 10 μg), amoxicillin-clavulanic acid (AMC, 30 (20:10) μg), cefepime (FEP, 30 μg), cefotaxime (CTX, 30 μg), ceftriaxone (CRO, 30 μg), cefuroxime (CXM, 30 μg), meropenem (MEM, 10 μg), ciprofloxacin (CIP, 5 μg), tetracycline (TE, 30 μg), chloramphenicol (C, 30 μg), gentamicin (CN, 10 μg), trimethoprim sulfamethoxazole (SXT, 25 (1.25: 23.75). The authors, might explain to PONE readers what the numbers with the μg units mean and why μg is missing for trimethoprim sulfamethoxazole.

8. Full legend information should accompany each table. For example, the authors should not leave readers to assume exactly what C1/C2 means. In Table 1, the value of the data within and outside brackets in each cell should be defined at the top of the column.

9. The authors should introduce a section on limitations encountered in this study. Limitations include lack of molecular studies to achieve source attribution for the bacteria isolated as well as determine dissemination pathways. Though the authors have tried to indirectly and subtly link the Rift Valley niche to AMR pathogens in these birds by mentioning environmental stewardship. Evidence for such is not presented in this study.

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what does this mean? ). If published, this will include your full peer review and any attached files.). If published, this will include your full peer review and any attached files.

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Reviewer #1: No

Reviewer #2: No

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A One Health Assessment of Antimicrobial-Resistant Enterobacterales in Migratory Little Stints (Calidris minuta) and Aquatic Ecosystems in the Kenyan Rift Valley

PONE-D-25-57615R1

Dear Dr. Mbuthia,

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.

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Kind regards,

Mabel Kamweli Aworh, DVM, MPH, PhD. FCVSN

Academic Editor

PLOS One

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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??>

Reviewer #1: Yes

Reviewer #2: (No Response)

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

Reviewer #1: Yes

Reviewer #2: (No Response)

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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: (No Response)

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

Reviewer #1: Yes

Reviewer #2: (No Response)

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Reviewer #1: Thank you for addressing my comments. A minor correction is required on lines 31–32 of the abstract, as content appears to be omitted between "lowest for" and "and meropenem."

Reviewer #2: "Since the main bodies of both lakes are unsuitable for hydration or domestic use due to their extreme chemistry, significant interaction between humans, livestock, and C. minuta occurs at the surrounding peripheral freshwater sources, including springs, rivers, marshes, and boreholes." This new statement added at Line 107 is welcomed. However, it might require a reference especially if the authors did not specifically observe what was stated.

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what does this mean? ). If published, this will include your full peer review and any attached files.). 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 For information about this choice, including consent withdrawal, please see our Privacy Policy .-->

Reviewer #1: No

Reviewer #2: No

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