PONE-D-23-42913Development and assessment of an immobilized bacterial alliance that efficiently degrades tylosin in wastewaterPLOS ONE

Dear Dr. Gao,

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.

Authors <word-sub class="revised" data-idx="0-0-0">are required</word-sub> to <word-sub class="revised" data-idx="0-0-1">assess</word-sub> and <word-sub class="revised" data-idx="0-0-2">implement</word-sub> the <word-sub class="revised" data-idx="0-0-3">essential revisions</word-sub> for a <word-sub class="revised" data-idx="0-0-4">subsequent</word-sub> round of <word-sub class="revised" data-idx="0-0-5">manuscript review.</word-sub> <word-sub class="revised" data-idx="0-1-0">The reviewers raised several concerns, such as</word-sub> the biosafety of the bacterial <word-sub class="revised" data-idx="0-1-2">strain</word-sub> and the efficiency of pollutant <word-sub class="revised" data-idx="0-1-5">species degradation.</word-sub>

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Academic Editor

PLOS ONE

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China Agriculture Research System of MOF and MARA (CARS-37)"

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China Agriculture Research System of MOF and MARA (CARS-37)"

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Reviewers' comments:

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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: Partly

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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: No

Reviewer #2: Yes

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

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Reviewer #1: Yes

Reviewer #2: No

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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: In their work, Zhao et al examined the possibility of using a consortium consisting of two newly isolated bacterial strains, Kurthia gibsonii (TYL-A1) and Klebsiella pneumonia (TYL-B2), for microbial degradation of tylosin. For better performance, the consortium was immobilized in PVA-SA-AC. Possible mechanisms for improving TYL biodegradation through immobilization are discussed. The feasibility of TYL removal was tested using real wastewater. Despite the overall good impression of the paper, I have some concerns, mainly about the real wastewater part and the pathogenicity, see comments below:

Specific comments:

Line 18: “PVA-SA-AC” should be defined upon first use

Lines 74-77: The poor sentence construction, please check

Line 79: what is TSM?

Lines 86-87: “from an agricultural environment.” What is the agricultural environment? Please be more specific when formulating the purpose of the work.

Line 121: “spresd” please check

Line 147: “0.22 mm membrane” please check the units are correct.

Line 186: “in wastewater TYL” change to "in wastewater containing TYL"

Lines 186-187: “standing wastewater and supernatant were” please give more detail what is standing wastewater and supernatant?

Line 188: “wastewater containing TYL” how much TYL is contained in wastewater?

Line 189: “NH+ 4−N” please change to “NH4+-N”. Please use the same format for other nitrate-N and phosphate-P

Line 246: what are CTC and OTC?

Line 261: “++++ softened…” please check, should be +++

Line 266: What does the word “concretization” mean here?

Line 279: “in contrast to inefficient degradation by free bacteria” degradation by free bacteria was not inefficient

Line 293: "a" and "b" are not shown on images

Line 315: “represent SEM of three experiments” please define SEM here

Line 315: “wastewater from aquaculture” Wastewater from aquaculture or agriculture? please clarify

Line 373: “Degradation efficiency was reduced, but it can be recycled.” the meaning is not clear, please rephrase

Line 376: “initial increase was attributed to TYL and immobilized carriers” please state your thesis more clearly. How could TYL and the immobilized carrier contribute to the increase in COD?

Lines 376-377: “whereas the decrease was attributed to degradation of TYL” what was the COD equivalent of TYL contained in wastewater?

Line 380: “probably due to bacteria utilizing P” Why was there a difference compared to the control?

Lines 380-382: not a logical explanation. First, ammonia levels generally do not increase during denitrification. Secondly, this is an aerobic process that does not favor denitrification.

General comment:

What about the safety concerns associated with strong pathogenicity of Klebsiella pneumonia? How does this relate to practical applicability? The propagation of an antibiotic-resistant pathogen is unlikely to be approved by the relevant authorities?

Reviewer #2: The review of the manuscript PONE-D-23-42913 has been completed. Concerns below needs to be addressed.

The abstract needs to be re-written to reflect the significance of the study.

Previous researches have reported the use of bacteria for the remediation of pollutant species. Authors should add some of these papers include their degradation efficiency

The study needs to be comprehensive. Adsorption accelerated oxidation and biodegradation of pollutant specie besides antibiotics should be added. Here are some references: https://doi.org/10.3390/ijms231810637, https://doi.org/10.3390/ijerph19169962, https://doi.org/10.3390/w14132063.

The novelty and significance of the study should be comprehensively presented.

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Reviewer #1: No

Reviewer #2: No

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PONE-D-23-42913R1Development and assessment of an immobilized bacterial alliance that efficiently degrades tylosin in wastewaterPLOS ONE

Dear Dr. Gao,

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.

The reviewers' comments need to be addressed, and the manuscript must be revised accordingly before it can be accepted for publication.

Please submit your revised manuscript by May 20 2024 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 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'.

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.

We look forward to receiving your revised manuscript.

Kind regards,

Marcos Pileggi, Ph.D

Academic Editor

PLOS ONE

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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

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

Reviewer #1: Yes

Reviewer #2: N/A

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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: Yes

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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: Yes

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: Thank you for addressing my comments.

Please

1) Lines 82-84: You should not give description of agricultural environment in the aim of the work. It can be indicated in the introduction above.

2) Lines 275-278: Just define SEM in parentheses rather than in a separate sentence, it's redundant.

3) Line 298: pH, not PH

4) Be clear about whether you are working with aquaculture or agricultural wastewater?

5) Lines 369-375: Do you mean that composite carbon is sourced from beads and those beads are being biodegraded? Because it seems that wastewater did not have undissolved organics.

Reviewer #2: The review of the revised manuscript titler'Development

and assessment of an immobilised bacterial alliance that effectively degrades tylosin in wastewater has been completed. The concern below is required to be addressed before manuscript can be accepted.

The manuscript has been improved comprehensively. However, before it can be accepted. Authors should address the minor comment below.

The Line 79-80 should be supported with a hypothesis to give deeper insight into the objective of the study.

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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: Yes: Abdulrahman Oyekanmi Adeleke

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While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Development and assessment of an immobilized bacterial alliance that efficiently degrades tylosin in wastewater

PONE-D-23-42913R2

Dear Dr. Gao,

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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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,

Marcos Pileggi, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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: (No Response)

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: (No Response)

Reviewer #2: Yes

**********

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

Reviewer #1: (No Response)

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: (No Response)

Reviewer #2: Yes

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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 Response)

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: (No Response)

Reviewer #2: Concerns has been comprehensively addressed. Manuscript is suitable for acceptance in the current form.

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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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