PONE-D-20-17828

Microbiological quality and genotoxicity of domestic water sources: A combined approach using Micro biological Survey method and mutagenesis assay (micronucleus test) in root tips of Vicia Faba in the West Region of Cameroon

PLOS ONE

Dear Dr. Mabvouna,

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.

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We look forward to receiving your revised manuscript.

Kind regards,

Swatantra Pratap Singh, Ph.D.

Academic Editor

PLOS ONE

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Additional Editor Comments (if provided):

The article titled “Microbiological quality and genotoxicity of domestic water sources: A combined approach using Micro biological Survey method and mutagenesis assay (micronucleus test) in root tips of Vicia Faba in the West Region of Cameroon” is interesting, however, it has very high similarity with a previous article on "Microbiological quality of water sources in the West region of Cameroon: quantitative detection of total coliforms using Micro Biological Survey method". This article cannot be accepted in the present form and need Major revision in terms of presentation, methodology, and discussion. The manuscript lacks many technical details that require clarification and/or improvement. Authors are suggested to address the reviews comments in order to make the manuscript suitable for publication and provide point-by-point justification or rebuttal against the comments:

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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: No

Reviewer #2: Partly

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

Reviewer #1: No

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: 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: No

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: The work has been found to be very interesting, however there are some major suggestions with the respect to the design of the experiment and also discussions in the writeup:

1) Details about the sampling to be given in more depth

2) As the microbial load of the water is to be tested, the time of the day or even the temperature at the time of collection is crucial, please provide the data

3)Why E.coli, please explain the need for ennumeration of the indicator, depth in literature lacking especially in the introduction and discussion

3) Why an enzymatic assay like MBS, why not membrane filtration? Explicitly mention the advantages and the underlying principle and the rationale behind the choice of the method

4)Micronucleus test is a standard method, please cite where it has been adapted from.

5)Flowchart on the protocol for micronucleus test is recommended

6)22 samples may be too few to come up with a valid conclusion

Reviewer #2: Manuscript Number: PONE-D-20-17828

Title: Microbiological quality and genotoxicity of domestic water sources: A combined approach using Micro biological Survey method and mutagenesis assay (micronucleus test) in root tips of Vicia Faba in the West Region of Cameroon

Authors: Rodrigue Biguioh Mabvouna, Sali Ben Béchir Adogaye, Patrick Martial Pete Nkamedjie, Adrillene Laure Wondeu Deutou, Martin Sanou Sobze, Jean Blaise Kemogne and Vittorio Colizzi

Article Type: Research Article

Journal: PLOS ONE

Comments: The manuscript highlights the poor quality of domestic water sources in the Central African sub-region. The study deals with the microbial and genetic toxicity induced factors responsible for deteriorating water quality of different natural/domestic sources of drinking water. While this is worthy of investigation, the manuscript lacks in clarity of presentation, especially the methodology and discussion section. The manuscript provides a good read on detection of total coliforms (TC) which was done using the Micro Biological Survey-HACCP & water Easy test along with micronucleus test to assess the genetic toxicity caused due to the poor water quality. However, the approach of the study lacks many technical details that need clarification and/or improvement. Authors should improve the usage of the English language while revising the manuscript. Authors are suggested to address following comments in order to make the manuscript suitable for publication and provide point-by-point justification or rebuttal against each comment:

1. The content of abstract requires modification. Authors are suggested to make the abstract more clear and informative. Provide concise and factual abstract briefly and sequentially stating the aims (objectives), major results, and principal conclusions of the work. Derive the conclusion at last. Provide the data (values) for significant findings of the present study.

2. Abstract, line 29, and Study design and period, line 123, At each site,…genotoxic analysis. The word ‘litter’ should be replaced by ‘litre’ which is a unit of volume.

3. Introduction, lines 78-80, According to World Health Organization…between life stages. Please rephrase the sentence.

4. Total coliforms detection, Lines 130-131, what does it mean by rapid color change? Specify the changes in color corresponding to a definite range for the determination of increasing or decreasing coliform count.

5. Total coliforms detection, Lines 130-131, which reagent is the author talking about? Please mention.

6. Total coliforms detection, Lines 141-145, please mention the specific color change and concentration range (very high, high and low) for coliform count after 24 hours. Also clarify the reason for the reducing coliform counts with the passage of time.

7. Total coliforms detection, lines 141-145, A color change…103 CFU/ml. Please rephrase the sentence.

8. Of the 4 water sources sampled (as mentioned in study design and discussion), results of drilling water is not mentioned in Table 1. Further, results lack the complete details on TC concentration range along all the sources. Also, provide the coordinates for the sampling locations.

9. The reason for opting/choosing Vicia faba as a study plant is not mentioned in the methodology. Authors need to explain with proper rationale or any background study.

10. MN (micronucleus) test abbreviation must be given in the sub headings rather than providing details within the text body for better understanding of readers. Also brief introduction about MN test is not provided as given for Micro Biological Survey-HACCP & water Easy test for coliforms. Please include.

11. The major criticism of the manuscript lies in the lack of technical details provided for the methodology adopted. Authors are suggested to make this section clearer and explanatory. How well water is supposed to be more contaminated with higher coliform count during rainy reason and not in the surface water source? Explanation provided is not satisfactory and subject to contradiction. More meaningful and concrete justification is required.

12. Discussion, TC concentration detected highest among wells but later genotoxicity effects has been reported in spring water, river water as well as well water. What could be the possible reason for such differences? Authors need to address the same.

13. What specific genotoxicity effects do author referring to?? Whatever genetic changes/ mutation detected in the experiments is nowhere mentioned in the manuscript. Please clarify.

14. Discussion, line 215, “However, the TC concentration of untreated source water was found to be higher than treated water sources with no significant difference” – the sentence itself showing significant differences in TC concentration. Please rephrase suitably.

15. Discussion, lines 217 – 219, (Other standardized……..environmental pollutions) – past studies and future need for more such studies needs to be differentiated well. Rephrase the sentence carefully.

16. Discussion, line 232, intensive agricultural activities instead of “intensives”. Please check the whole manuscript for other grammatical errors.

17. The conclusion section must state the key findings of the study. Authors are suggested to draw major inferences/primary conclusions first from the results followed by the secondary conclusions reached through the critical analysis.

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6. 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

[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.]

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.

PONE-D-20-17828R1

Microbiological quality and genotoxicity of domestic water sources: A combined approach using Micro biological Survey method and mutagenesis assay (micronucleus test) in root tips of Vicia Faba in the West Region of Cameroon

PLOS ONE

Dear Dr. Mabvouna,

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.

Please submit your revised manuscript by 15th Dec 2020. 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Swatantra Pratap Singh, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

The article has been improved, however few miner corrections are required before publication as suggested by the reviewers.

[Note: HTML markup is below. Please do not edit.]

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: Partly

Reviewer #2: Yes

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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: No

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: 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: The work done is interesting however some suggestions on improving the clarity of the writeup is recommended:

Line 130: 1 mL of what concentration of thiosulphate? Is it sodium thiosulphate, if yes please specify, if not please mention the salt name explicitly

Line 133: Mention explicitly the temperature and pH were taken before or after placing the samples in the ice box

Table 1: The temperature mentioned is for the water or ambient air temperature? Also, mentioning the time of collection is suggested

Line 160-164: Add the reference

For the section of total coliform detection: The test details insufficient, which enzymes play a crucial role in the color change? More details on the mechanism of color change needs to be explained to understand how the kit works?

Line 219: Why so much variation in the pH, was it because the samples were collected at different time frames of the day, was the time of collection not kept constant? Or was there appearance or disappearance of algal blooms?

Reviewer #2: The manuscript has been revised in accordance with the comments provided. Authors have also satisfactorily answered and clarified the doubts raised. The revised manuscript is now suitable for acceptance and publication in the Journal.

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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

[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.]

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.

Microbiological quality and genotoxicity of domestic water sources: A combined approach using Micro biological Survey method and mutagenesis assay (micronucleus test) in root tips of Vicia Faba in the West Region of Cameroon

PONE-D-20-17828R2

Dear Dr. Mabvouna,

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 for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

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,

Swatantra Pratap Singh, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

As per reviewers comments and we please to accept the article for the publication.

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

**********

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: 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: The paper has covered a very important topic and the authors have tried and included all the revisions suggested by the reviewers

Reviewer #2: The manuscript has been revised suitably. The revised manuscript is now suitable for acceptance and publication in the Journal.

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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