PONE-D-22-25657Biogenic sunflower oil-chitosan decorated fly ash nanocomposite film using white shrimp shell waste: antibacterial and immunomodulatory potentialPLOS ONE

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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: Partly

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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: Authors have taken up an important topic for their research, the article is well supported by experimental procedures and characterization. However, there are few points which needs to be addressed. My concerns are as follows.

1. The abstract should be precise and more informative.

2. Introduction 3rd paragraphs... Another study conducted by Mitiko. Researcher’s name is underlined for any specific reason or it is a typo?

3. Authors mentioned two reports on chitosan loaded fly ash nanocomposite, they should mention also mention how their procedure are different from previously reported one.

4. If possible introduction should be shortened and the references be reduced.

5. Section: Isolation of chitin and formation of chitosan: Authors developed their own demineralization step or adopted previously reported procedure?

6. Section: Isolation of chitin and formation of chitosan: what is the purpose of bleaching the samples?

7. Section: Isolation of chitin and formation of chitosan: Is there any other reported procedure for deproteinization ?

8. What is the significance of Cell viability assay?

9. “RT-PCR” full form should be mentioned at first place of appearance.

Reviewer #2: Manuscript ID: PONE-D-22-25657

Title: Biogenic sunflower oil-chitosan decorated fly ash nanocomposite film using white shrimp shell waste: antibacterial and immunomodulatory potential

Journal: PLOS ONE

A new sunflower oil-chitosan decorated fly ash (sunflower oil/FA-CSNPs) bionanocomposite film was synthesized using the extract of Litopenaeus vannamei (White shrimp) and monitoring as an antibacterial and immunomodulatory agent. The chitosan (CS) was prepared by deacetylation of chitin isolated from white shrimp extract. Fly ash-chitosan nanoparticles were produced by using glacial acetic acid and sodium tripolyphosphate solution as cross-linkage. The ultrafine polymeric sunflower oil-CS film was fabricated by dissolving CS in glacial acetic acid for 24 h with continuous stirring. The nanostructure of the fabricated polymeric film was confirmed and characterized by different microscopic and spectroscopic approaches including, FTIR, XRD, and SEM. The results are well supported by the conclusion. I recommend major revision of the manuscript before it can be accepted. I request the authors at addressing all comments and suggestions listed below.

Comments and suggestions:

1. “The formed sunflower oil/FA-CSNPs bionanocomposite film showed promising antibacterial and immunomodulatory potentials towards Bacillus subtilis, Pseudomonas aeruginosa and macrophage-derived RAW264.7 cell line, respectively” – Mention the results here.

2. “Keywords: Litopenaeus vannamei; chitin, chitosan; fly ash; antibacterial potential, immunomodulatory effect."- Use different keywords, which are not used in title.

3. “Recently, the control of organic waste disposal is a growing issue

for the legislation and environmental safety..”--- I suggest the author, to discuss a paragraph related to pollution due to the presence of different contaminants. The authors are recommended to check the below related references, which will improve the supporting information.

Journal of Molecular Liquids 317, 2020, 113916

Separation Science and Technology 55 (10), 2020, 1766-1775

Journal of Cleaner Production 156, 2017, 426-436

Journal of Hazardous Materials 400, 2020, 123247

4. Also in recent times, there has been a great incentive for generating environmentally friendly materials for water purification and wastewater treatment, which need to be included in introduction section.

Photochemistry and photobiology 94 (5), 2018, 935-941

Nanomaterials 9 (5), 2019, 776

Arabian Journal of Chemistry 12 (5), 2019, 633-651

Environmental research 2019, 170, 389-397

5. “White shrimp (L. vannamei) shells were purchased from a local market in Riyadh, Saudi

Arabia.”-what basic criterion were chosen for shrimp collections?

6. “Figure 1: FTIR spectra of (a) Fly ash, (b) chitosan, (c) FA-CSNPs, and (d) Sunflower oil/FACSNPs-bionanocomposite film measured at wavenumber 4400-400 cm-1”. mention the optimal conditions in each figure, while needed.

7 “The dried homogenized shrimp shells (100 g) were demineralized (removing calcium carbonate

and phosphate) by adding 1.0 mol L-1 hydrochloric acid (1L) at room temperature under

agitation for 12 h at 250 rpm.”- Does the author followed any reported work, if yes then the novelty needs to be discussed?

8. “The FTIR spectrum of raw fly ash exhibited broad absorption peaks at 3449 and 1634 cm-1 assigned to O-H and H-O-H groups stretching and deformation vibrations of the water molecules” - Need supporting information, Journal of environmental management 219, 2018, 285-293

9. “The crystalline size of FA-CSNPs was found to be 6.90 nm when calculated using Scherrer’s equation:” - Desalination and Water Treatment 57 (46), 2016, 21863-21869

10. “The impact of FA, FA-CSNPs, and bionanocomposite film on the shape of both treated and untreated B. subtilis and P. aeruginosa were examined using a scanning electron microscope (SEM).”- Provide the details of the manufacturer and also do same for all instrument used including city and country in experimental section.

11. The English quality not up to the mark. All the typos and grammar need to check thoroughly in the manuscript.

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Reviewer #1: No

Reviewer #2: No

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Biogenic sunflower oil-chitosan decorated fly ash nanocomposite film using white shrimp shell waste: antibacterial and immunomodulatory potential

PONE-D-22-25657R1

Dear Dr. Amina,

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,

Sivasankar Koppala

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: 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: N/A

Reviewer #2: Yes

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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: Authors have made all the required corrections satisfactorily.

Reviewer #2: All the comments raised by me are addressed properly. I suggest acceptance of the paper 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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