PONE-D-22-31808BIOPROSPECTION OF THE TRICHOMONACIDAL ACTIVITY OF LIPID EXTRACTS DERIVED AT DIFFERENT STAGES OF DEVELOPMENT FROM Gigartina skottsbergii AGAINST Trichomonas vaginalisPLOS ONE

Dear Dr. Borsuk,

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Academic Editor

PLOS ONE

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Additional Editor Comments:

Authors must improve the discussion by comparing the efficacy of the fatty acid rich extracts with other extracts that are available in the natural source used. Author may follow and cite Doi: 10.2174/1389557516666151120121036.

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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: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: N/A

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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: Yes

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: The manuscript entitled “BIOPROSPECTION OF THE TRICHOMONACIDAL ACTIVITY OF LIPID EXTRACTS DERIVED AT DIFFERENT STAGES OF DEVELOPMENT FROM Gigartina skottsbergii AGAINST Trichomonas vaginalis” is a nice piece of work describing the in vitro and in silico anti-parasitic activity of Gigartina skottsbergii against Trichomonas vaginalis. But before it can be accepted for publication in this high quality journal the following issues are needed to be addressed properly

1. Please provide a graphical abstract with brief caption

2. It remains elusive how valid the results from the shown experiments are. How many replicates were performed for each culture condition? So the description and interpretation of the data should be done more carefully.

3. Indicate the numbers of parasites as replicate for each experiment in the methodology section or figure legend.

4. The structural or ultra-structural changes in cell morphology after exposure to Gigartina skottsbergii extract should be studied.

5. Abbreviations in the picture should be explained.

Reviewer #2: Authors tried to established the trichomonacidal activity of fatty acids derived from the marine macroalgae Gigartina skottsbergii . Overall the article is interesting and can be published after some minor revisions as follows:

General Comments:

1. Several abbreviations are used in the text without full form. One time mention of the full form of the used abbreviations is required for general readers

2. Though overall language is good, but there is a scope of improving the standard of language. In some cases it is too wordy and unnecessary!

3. Images/ Graphs can be more attractive and colourful.

Specific Comments

Title:

It is too wordy! It can be shortened like “Bioprospection of the trichomonacidal activity of lipid extracts derived from marine macroalgae Gigartina skottsbergii”

Abstract:

Line 31: Trichomonas vaginalis should be italic.

Line 34, 39: in vitro and in silico should be italic and the pattern should be maintained all through the manuscripts

Line 34-35: ‘marine microalgae’ should be placed before ‘Gigartina skottsbergii’

Introduction:

Line 74: ‘….considered important considering …….’ Rewrite the sentence.

Line 78: Instead of ‘non-viral infections’ write ‘protozoan infection’

Line 79: Add ‘particularly in female’ after ‘genitourinary system’

Material and Methods:

Macroalgae Specimens

Line 93-99: How the specimen is identified? What is its accession no.? Necessary information should be provided

Extraction and Derivatization

Line 103: Why the fatty acids are only extracted? Are there any clues for using fatty acids as anti-protozoan drug? Because alga produces an array of secondary metabolites such as alkaloids, flavonoids, glycosides, terpenoids, and phenazines which have alleged medicinal properties!

Line 103: Bligh and Dyer (B&D) - year and reference no. is missing

Line 110: ‘described by [15]’- author is missing!

Instrumentation and Quantification

Line 130-134: ‘Methanol, chloroform, …………..purchased from Supelco (Bellefonte, USA).’ Unnecessary and delete from the text.

Line 141: Full form of ‘TvMGL, TvPNP, TvTPI, and TvLDH’ should be given

Line 145-46: ‘Linker structures ….. PubChem program’.- unnecessary and delete

Line 149,152: ‘Autodock’ should be ‘AutoDock’

Parasite culture conditions

Line 166: Full form of ‘GFG, GFT, and GF’ should be given

Line 166: Correct the information ‘Sena-Lopes (et al., 2017)’- et al. should be italic and outside the parenthesis, also provide the reference no. within [ x]

Line 173: ‘2.6 × 105’ should be ‘2.6 × 105’

Line 180: ‘CO2’ should be ‘CO2’

Cytotoxicity Assay

Line 203: Navarrete-Vázquez (et al., 203 2015) - et al. should be italic and outside the parenthesis, also provide the reference no. within [ x]

Gene expression analysis of T. vaginalis

Line 237-238: ‘Forward5'-CG….AGG3'andReverse 5'-TCCT…..AAC-3'- provide space in appropriate places!

Results:

Anti-T. vaginalis Activity

Line 277: 89,61% and 86,95%, should be 89.61% and 86.95%, respectively

Molecular docking

Line 331: Required space is missing in ‘(∆Gbinding)’

DISCUSSION

Line 384: Few references are required for the comment “Several of the identified constituent compounds are already reported in the literature…”

Line 398-399: ‘Therefore, it was established that the evaluated extracts exhibited…’ this sentence is not in conformity with the previous sentence. Rewrite accordingly

Line 402: ‘Bonde (et al., 2021)’- et al. should be italic and outside the parenthesis, also provide the reference no. within [ x]

Line 448-449: ‘(LEITSCH; KOLARICH; DUCHÊNE, 2010; JONES et 449 al., 2016)’- it is not the Plos Ones’ reference style!

Conclusion:

Line 478: Write ‘Gigartina skottsbergii’ after ‘ subantarctic marine algae’

Line 479: ‘T. vaginalis’ should be italic

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Reviewer #1: Yes: Dr. Priya Roy

Reviewer #2: No

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Attachments

Attachment

Submitted filename: review report of PONE-D-22-31808.docx

BIOPROSPECTION OF THE TRICHOMONACIDAL ACTIVITY OF LIPID EXTRACTS DERIVED FROM MARINE MACROALGAE Gigartina skottsbergii

PONE-D-22-31808R1

Dear Dr. Borsuk,

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,

Suprabhat Mukherjee, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Authors have addressed all the comments.

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

**********

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

**********

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: Authors are successfully able to comply all the questions raised by the reviewers. They should check the ms inconformity with PlosOne guidelines.

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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 #2: Yes: Biplob Kumar Modak

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