PONE-D-24-45060Mangrove consortium resistant to the emerging contaminant DEHP: Composition, diversity, and ecological function of bacteriaPLOS ONE

Dear Dr. Natascha Krepsky,

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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: Yes

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: Yes

Reviewer #2: No

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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: No

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: Comments to the authors:

In this manuscript, MP001 communities were exposed to DEHP at different concentrations under laboratory conditions, and high-throughput sequencing techniques were used to assess bacterial composition, diversity, and potential ecological function. The resistance, composition, diversity and ecological function of MP001 to the emerging pollutant DEHP were evaluated. Studies have shown that the MP001 community shows potential as an eco-friendly bioreactor for DEHP. But there are some problems with this manuscript: First, there are some problems with structure and English grammar. Second, you need to provide detailed information about the laboratory equipment and application. Finally, issues related to community function and community diversity warrant further analysis and consideration of possible environmental influences on community structure and function. The specific comments are provided below.

Specific comments:

1. The section "1. Introduction" can add research hypothesis, deeper research purpose and research significance

2. In section "2.2 Sampling and consortium isolation", it is recommended to draw a geographic map of the sample area.

3. In the section "2.2 Sampling and consortium isolation", to what depth was the sediment collected? Are they randomly selected sediments or are they collected at specific locations?

4. In the section "2.3. Bacterial inoculum preparation", please provide the manufacturing information of each experimental instrument.

5. In the section "2. Materials and Methods", I noticed that you used some applications to analyze the data, please provide their version information.

6. Line 156. There is a grammatical error that suggests adding the indefinite article "a" between "as" and "negative".

7. In the section "3. Results", whether attention was paid to the possible effects of other environmental factors (such as temperature, pH, salinity, etc.) on community structure and function.

8. In "3.2.2. Diversity of the consortium MP001 before and after DEHP exposure", there was no significant change in community diversity after DEHP treatment. But it is worth exploring in depth whether this means that the community's resistance to DEHP is due to its inherent diversity or due to other factors.

9. . In the section "3.2.3. Potential ecological function of the consortium MP001 before and after DEHP exposure", although fermentation is mentioned as the main potential ecological function, Other possible ecological functions or interactions among community members were not further explored.

10. It is suggested to present the "4. Discussion" part in subsections.

Reviewer #2: Comments:

In this study, the authors isolated a bacterial consortium from neotropical mangrove using Di(2-ethylhexyl) phthalate (DEHP) as a stress source. They aimed to identify and characterize the DEHP-resistant microorganisms through gradient concentration exposure, DNA sequencing. The findings demonstrated that MP001 possesses a high potential for application in bioremediation purposes. However, significant concerns exists regarding the environmental significance and reasonability of the exposure design in its current form. Additionally, there are issues with the layout of figures and redundant/inaccurate information conveyed in some figures, which greatly compromises the readability of this paper.

The following specific points require further clarification and addressing:

1.In the Abstract, the authors mentioned the bacterial composition of exposure treatment and negative control. Are there any conclusion based on the changes in treatment and negative control groups?

2.The background and environmental context in which the study was conducted lack clarity. The selection of exposure doses ranged from 0.05 to 6 mg/L was based on specific considerations. However, there is a lack of available data or descriptions regarding the actual range of DEHP concentrations in the environment, which forms the core focus of this research.

3.In the present study, MP001 was isolated from the sediment of the Magé mangrove incubated at 37 ℃, and the exposure experiment was carried out at 35 ℃ ± 1. However, the average temperature in Rio de Janeiro state was reported as 17-36 ℃. Why did the authors choose such high temperatures? Does it really make sense to conduct bacteria isolation and exposure at these temperature?

4.In this study, the sediment sample was collected from the Magé mangrove in Rio de Janeiro state, which seems that the sediment sample was sampled at only one sampling site in the Magé mangrove. I’m worrying about the representativeness of this study because of the low sample size. If not, please provide the sampling sites of the study.

5.Could the media used for the separation and culture have a certain effect on bacteria determination? Why the media employed to bacteria isolation and exposure experiment were different?

6.The authors demonstrated that DEHP could provide as a carbon resource and degraded by the MP001 consortium in Line 313-314, Page 13. However, DEHP concentrations after exposure were didn’t determine. Thus, I consider this conclusion is unreasonable.

7.In Discussion section, the authors explain the dominant role of ARGs in xenobiotics resistance. However, it seems that the ARGs identification was absent in the present study.Therefore, I consider many elements of these paragraphs to be redundant and irrelevant to the content of this paper.

8.Ensure consistency in the significance in italics across the manuscript. Besides, several mistakes, such as “DEH Psediment” in Line 352, Page 14, disordered graphs (Figure 2 and 3)......

9.The authors should check and correct the mistakes in references according to the format requirement of the journal.

10.Several journal abbreviations need to be fixed and standard journal title abbreviations should be used throughout.

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Reviewer #1: No

Reviewer #2: No

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Mangrove consortium resistant to the emerging contaminant DEHP: Composition, diversity, and ecological function of bacteria

PONE-D-24-45060R1

Dear Dr. Natascha Krepsky,

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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Kind regards,

Mei Li

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 #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 #2: Yes

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

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 #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 #2: Yes

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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 #2: (No Response)

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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

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Submitted filename: Comments.docx