PONE-D-21-17789

Bioengineered phytomolecules-capped silver nanoparticles using Carissa Carandas leaf extract to embed on to urinary catheter to combat UTI pathogens

PLOS ONE

Dear Dr. Saravanan,

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 Aug 13 2021 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: http://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,

Amitava Mukherjee, ME, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. Please include your tables as part of your main manuscript and remove the individual files.

3. Thank you for stating the following financial disclosure:

 “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”

At this time, please address the following queries:

a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution.

b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”

c) If any authors received a salary from any of your funders, please state which authors and which funders.

d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.”

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

4. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. PLOS defines a study's minimal data set as the underlying data used to reach the conclusions drawn in the manuscript and any additional data required to replicate the reported study findings in their entirety. All PLOS journals require that the minimal data set be made fully available. For more information about our data policy, please see http://journals.plos.org/plosone/s/data-availability.

Upon re-submitting your revised manuscript, please upload your study’s minimal underlying data set as either Supporting Information files or to a stable, public repository and include the relevant URLs, DOIs, or accession numbers within your revised cover letter. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. Any potentially identifying patient information must be fully anonymized.

Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access.

We will update your Data Availability statement to reflect the information you provide in your cover letter.

5. We note that Figures 8, 10 and 11 in your submission contain copyrighted images. All PLOS content is published under the Creative Commons Attribution License (CC BY 4.0), which means that the manuscript, images, and Supporting Information files will be freely available online, and any third party is permitted to access, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution. For more information, see our copyright guidelines: http://journals.plos.org/plosone/s/licenses-and-copyright.

We require you to either (1) present written permission from the copyright holder to publish these figures specifically under the CC BY 4.0 license, or (2) remove the figures from your submission:

a. You may seek permission from the original copyright holder of Figures 8, 10 and 11 to publish the content specifically under the CC BY 4.0 license.

We recommend that you contact the original copyright holder with the Content Permission Form (http://journals.plos.org/plosone/s/file?id=7c09/content-permission-form.pdf) and the following text:

“I request permission for the open-access journal PLOS ONE to publish XXX under the Creative Commons Attribution License (CCAL) CC BY 4.0 (http://creativecommons.org/licenses/by/4.0/). Please be aware that this license allows unrestricted use and distribution, even commercially, by third parties. Please reply and provide explicit written permission to publish XXX under a CC BY license and complete the attached form.”

Please upload the completed Content Permission Form or other proof of granted permissions as an "Other" file with your submission.

In the figure caption of the copyrighted figure, please include the following text: “Reprinted from [ref] under a CC BY license, with permission from [name of publisher], original copyright [original copyright year].”

b. If you are unable to obtain permission from the original copyright holder to publish these figures under the CC BY 4.0 license or if the copyright holder’s requirements are incompatible with the CC BY 4.0 license, please either i) remove the figure or ii) supply a replacement figure that complies with the CC BY 4.0 license. Please check copyright information on all replacement figures and update the figure caption with source information. If applicable, please specify in the figure caption text when a figure is similar but not identical to the original image and is therefore for illustrative purposes only.

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

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: Partly

Reviewer #2: Partly

Reviewer #3: Partly

**********

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

Reviewer #1: Yes

Reviewer #2: N/A

Reviewer #3: N/A

**********

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

Reviewer #3: Yes

**********

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

Reviewer #3: No

**********

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: 1- Throughout the manuscript, there are a lot of typical, grammatical and unclear sentences like (Scherrs formula - AgNo3- and "Among the inorganic nanoparticles Silver nanoparticles (Ag NPs) considered to be a much more attention in scientific field", that lead to misunderstanding or confusion, I recommend sending this manuscript to a professional English language editing service.

2- In the introduction, authors should justify why they decided to use Ag NPs and leaves of C. carandas? highlight their advantages, because we can not simply use something because just it is available !

3- Line 60, "Leaves of C. carandas were used to yield Ag NPs", I think you need to rephrase this sentence, as leaf extract can only be used to stabilize formed Ag NPs and / or reduce the precursor solution of silver nitrate into Ag NPs.

4- Line 93, wavelength of Cu-Kα radiation is not correct, the correct value is 1.5406 Å.

5- In line 225, authors used Scherrer formula to determine crystalline size, and they mentioned non-correct wavelength in Line 93, then accordingly, the calculated size will not be correct. Please check this size again.

6- XRD pattern contains non-assigned peaks, please explain.

7- on FTIR spectra, it is better to highlight, peaks confirming the conjugation between Ag NPs and the extract.

8- On SAED pattern, you should assign the crystalline planes and match them with those obtained by XRD.

9- Fig. 2 is not clear, it is better to draw the data using suitable software !

10- Fig. 3 it is hard to see the label, also indicate the ZOI on the figure for each tested sample.

11- Fig.4, error bars should be added.

12- On Fig. 9, assign Ag NPs.

Reviewer #2: This work is having potential data, but no novelty, a simple repeat of already exiting report. The synthesis of AgNPs with plant extract is mushroomed in the literature. The MS, though, having good data, but I could not see any novelty to the field. I would suggest the work shall be modified to focus on Cather biofilm inhibition with standard drugs and other available AgNPs (might be synthesized by different methods).

Further

1. The Fig 10 is inappropriate, require evidence based pathway

2. Light Microscopy and Florescent Microscopy images shall be placed under suppl doc

3. Include CFLSM image for biofilm inhibition

4. TEM is showing a cluster of AgNPs, required scale marked particles

5. Self agglomeration of synthesized AgNPs on storage is required

6. Language and presentation require editing e.g. In the Introduction Pseudomonas is written as Pseudomon as

Reviewer #3: I consider the manuscript is tecnically sound, however some conclusions and discussion must be reconsidered in order to be supported by data obtained, I believe that description of results, the discussion and conclusions are highly restricted by language and strongly suggest a revision by professional editing service.

Detailed minor and major revisions are yellow highlighted in manuscript file attached to revision. in general minor and major revisions are:

Bioengineered phytomolecules-capped silver nanoparticles using Carissa Carandas leaf extract to embed on to urinary catheter to combat UTI pathogens

Minor revisions

All minor revisions are highlighted in manuscript file, these include suggestion for rewrite sentences, and simple changes.

Major revisions

Abstract and introduction

Grammar revision is suggested in some parts of these sections, in manuscript file are highlighted in yellow.

Material and methods

Grammar revision is suggested in some parts of this section, in manuscript file are highlighted in yellow.

Synthesis and optimization of AgNPs production

Include units of Ag ion concentration, volume of leaf extract, etc.

Antibacterial activity

I suggest modification of titles and subtitles order, and include some methodology description described in other method section.

Include description about how the AgNPs concentration was calculated.

Biofilm inhibition assay

Indicate concentration of AgNPs in concentration units (i.e. mg/L) instead of volume units. If cocnetration and volume of AgNps are equivalent please indicate and explain

In Section 2.12 it is not clear the objective of this experiment, please justify.

Results

I suggest to maintain the same subtitles used in methods section in order to establish an order and accordance between methods and results

I suggest include images of AgNPs suspensions obtained at different synthesis conditions (i.e. varying pH, leaf extract concentration, time reaction and Ag ions concentration)

I consider it is necessary to provide clear description of parameters used in each optimization condition of results obtained and presented in fig 1.

I considered necessary to clearly indicate which are the optimal parameters selected for AgNPs synthesis and criteria used for the establishment of these parameters.

It is not clear how the average size of AgNPs observed by HR-TEM was calculate, please include description.

I suggest to include information about how the MICs were calculated?

The fig 4 shows an important inhibition of bacterial growth (O.D.) at 160 mg/L however higher concentration must be proved in order to establish the MICs. I suggest include O.D. measurements of cultures exposed to higher concentrations of AgNPs to obtain a 100% of growth inhibition and establish the MICs

Description of results obtained by SEM must be wide described based on the results presentes in figure 9.

I suggest that the section of results 3.10 (Mechanisms of antibacterial and antibiofilm activity of AgNPs) must be eliminated and included and well describer in discussion section.

Discussion

I suggest general revision of grammar of this sections, some parts of the text are not understandable. (yellow highlighted)

Lines 328-329

Question: With SPR intensity do you refer to intensity in colour? or intensity of the peak absorption in spectra? if you refer to the color, you must provide the images of AgNPs suspensions . if you refer to the absorption peak, in figure 1a a variation of peak intensity and wavelenght of maximum absorption was clearly observed, thus an effect of pH in the intensity of absorption peak is produced.

Lines 327-340

I consider that based on FTIR results, probable phytomolecules involved in stabilization and capping of AgNPs must be provided and make a comparison with results obtained in previous studies on which phytosynthesis of AgNPs was carried out.

Lines 346-347

I consider is important to indicate how the particle size average was determined, HR-TEM indicate certain grade of heterogenicity of particle size, and in this part of discussion you describe that AgNPs are homogeneous in size, however in conclusion section a size heterogeneity of AgNPs was mentioned. Please describe results, discussion and conclusion according to the data obtained.

Line 358

I consider that a wide discusion based on the scientific litterature about the effciency of AgNPs coated catheters against UTIs must be provided.

Line 378

I consider that a wide description of the figure 10 was necessary, adapt the information provided below to the mechanisms described in figure.

Line 386-394

I consider that this part of discussion must include comparison of the previous studies described with the results obtained in this work. And include a wide discussion about phytomolecules involved in AgNPs synthesis.

Conclusions

I suggest rewrite the conclusions, cause I consider that some conclusions show discrepancy with the results and discussion, some of this conclusions are not supported by data presented.

Figures and tables

In general I suggest to improve the figure description, in order to be clear, informative and to support the description of the results. Also improve of resolution is recommended.

**********

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: Yes: Mohamed Abd Elkodous

Reviewer #2: No

Reviewer #3: Yes: LUZ ELENA VIDALES RODRIGUEZ

[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.

Attachments

Attachment

Submitted filename: PONE-D-21-17789_reviewer.pdf

PONE-D-21-17789R1

Bioengineered phytomolecules-capped silver nanoparticles using Carissa carandas leaf extract to embed on to urinary catheter to combat UTI pathogens

PLOS ONE

Dear Dr. Saravanan,

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.

==============================

ACADEMIC EDITOR: Your manuscript can be accepted provided you are ready to undertake minor revision as suggested by reviewer 3.

==============================

Please submit your revised manuscript by Sep 23 2021 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: http://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,

Amitava Mukherjee, ME, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

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.

[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 #3: 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 #3: Yes

**********

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

Reviewer #1: Yes

Reviewer #3: N/A

**********

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 #3: 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 #3: 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: In the revised version of their manuscript, authors addressed the required comments properly and I think the article is acceptable.

Reviewer #3: Comments has been adressed by authors, however, some mistakes remains in the manuscript, most of those are simple mistakes and easy to correct and has been highlighted in the manuscript file.

Description of an specific part in the discussion section (highlighted in mauscript file) can be improved.

In fig 11, I stronglly suggest to modify the figure, specifically, eliminate some cellular organelles which are specific for eukaryiotic cells.

**********

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: Yes: Mohamed Abd Elkodous

Reviewer #3: 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.

Attachments

Attachment

Submitted filename: PONE-D-21-17789_R1.pdf

Bioengineered phytomolecules-capped silver nanoparticles using Carissa carandas leaf extract to embed on to urinary catheter to combat UTI pathogens

PONE-D-21-17789R2

Dear Dr. Saravanan,

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,

Amitava Mukherjee, ME, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: