Biological Activities of Chamaecyparis lawsoniana (A.Murray bis) Parl. and Thuja plicata Donn ex D.Don Essential Oils: Toxicity, Genotoxicity, Antigenotoxicity, and Antimicrobial Activity

1. Summary

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in the re-submitted files.

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Yes

Many thanks for evaluation

Are all the cited references relevant to the research?

Yes

Many thanks for evaluation

Is the research design appropriate?

Yes

Many thanks for evaluation

Are the methods adequately described?

Yes

Many thanks for evaluation

Are the results clearly presented?

Yes

Many thanks for evaluation

Are the conclusions supported by the results?

Yes

Many thanks for evaluation

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: The authors have reported biological active experiments using essential oils derived from trees in the Cypress family. The description of experimental details is sound.

Response 1: Thank you very much for this comment.

Comments 2: Section 2. Materials and Methods

Section 2.1. Did the authors collect voucher samples of the plant materials? These should have been collected and the plant material should have been identified by a registered botanist.

Response 2: Thank you for pointing this out. We agree with this comment. In the originally submitted version, we omitted to state these informations. Therefore, in proof version, we have added details about voucher samples and the name of coauthor who has identified plant material. Please find suggested changes on page 2., Section 2., Subsection 2.1.; line numbers 83-86, marked in yellow.

Comments 3: Section 2. Materials and Methods

Section 2.2. Please state the origin/identification of D. melanogaster.

Response 3: Thank you for pointing this out. We agree with this comment. In the originally submitted version, we omitted to state that information. Therefore, we have added details about origin/identification of the D. melanogaster. Please find completed sentence on page 3., Section 2., Subsection 2.2., subsubsection 2.2.1, line numbers 94-95, marked in yellow.

Comments 4: Section 2. Materials and Methods

Section 2.3. Please state the origin/identification of A. salina.

Response 4: Thank you for pointing this out. We agree with this comment. In the originally submitted version, we omitted to state that information. Therefore, we have added details about the origin/identification of the A. salina. Please find rearranged and completed sentence on page,4., Section 2. Materials and Methods, subsection 2.3, line numbers 147-148, marked in yellow.

Comments 5: Was a GC analysis performed on the essential oil samples used in this study, or did they base their conclusions on the profile published earlier by one of the co-authors (Nikolić)? In practice, it is preferred that the authors perform the chromatographic analysis, determine the composition, and then proceed with the conclusions instead of assuming the profiles are identical.

Response 5: Thank you for this observation. GC analysis of the essential oils, that was used in presented manuscript, was performed, and published earlier by Nikolić et al., 2022. Considering that we used identical essential oils in the presented work, we did not perform GC analysis again on the same oils. So, at the beginning of the section Discussion, we rephrased the first sentence to clarify this. Please find this in the beginning of 4. Discusssion, page, 9., line numbers 350-352, marked in yellow.

Point 1: “The quality of the English language could be improved; however, the current form does present a distraction from the results and discussion within the paper.”

Minor editing of English language required.

Response 1:  Thank you for pointing this out. Native English speaker has revised our manuscript and has improved the English language. We have implemented corrections that we received from the English native speaker. Please find that corrections through the proved manuscript, marked in blue. Please note that all corrections in the manuscript related to English language did not change the essence of the text.

5. Additional clarifications

N/A

Response to Reviewer 2 Comments

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in the re-submitted files.

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Yes

Many thanks for evaluation

Are all the cited references relevant to the research?

Yes

Many thanks for evaluation

Is the research design appropriate?

Yes

Many thanks for evaluation

Are the methods adequately described?

Yes

Many thanks for evaluation

Are the results clearly presented?

Yes

Many thanks for evaluation

Are the conclusions supported by the results?

Yes

Many thanks for evaluation

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: The present research evaluates the Biological activities of Chamaecyparis lawsoniana and Thuja plicata essential oils.

Response 1: We agree with this comment.

Comments 2: The abstract reflects the main results of this research.

Response 1: Many thanks for the comment.

Comments 3: The introduction provides a good overview of the literature that exists on the topic and introduction with good arguments the objectives of the document.

Response 3: Many thanks for the comment.

Comments 4: The material and methods is well detailed and allows a perfect comprehension of how it was conducted.

Response 4: Many thanks for the comment.

Comments 5: The result section is clear, and tables and figures are of good quality, only minor details should be reviewed on this point, please see document attached.

Response 5: Thank you very much for pointing this out. We agree with this comment. Therefore, we have corrected/added details according to the request. Please find these changes, marked in yellow within revised manuscript, in Results section:

-on page 5., please note the changes in heading of Table 1, changed line 222, in Table 1, in seven columns of Table 2 (between lines 222 and 223), and consequently in the text, lines: 205, 206, 211, 212, and 221, all marked in yellow.

-on page 6., please note the changes in heading of Table 2, line number 248, in seven columns of Table 2 (between lines 248 and 249), and consequently in the footnote of Table 2, line 253, and consequently in the text the text, lines: 269, 270 and 271, all marked in yellow.

-on page 7., in Table 3, in first two columns between lines 291 and 292, as well as in footnote of the same table, all marked in yellow.  

-on page 8., in Figure 1 caption, line number 314, and in in Table 4 footnote, line number 330, marked in yellow. 

-on page 9., in Table 5 caption, lines 347 and 348, as well as in the beginning of Section 4, Discussion, lines 350-352, marked in yellow. 

Comments 6: The discussion is well organized and really complete.

Response 6: Many thanks for the comment.

Comments 7: The conclusion is in accordance with the results shown.

Response 7: Many thanks for the comment.

Point 1: English language fine. No issues detected.

Response 1:  Many thanks.

5. Additional clarifications

There are a few demands for correction of the technical errors that we have found within the provided PDF provided by Reviewer 2. Please find corrections, marked with yellow within revised manuscript, according to that demands at:

-page 4., subsection 2.5.2, the first paragraph; line number 183, marked in yellow.   

-page 10., subsection 4.1, the second paragraph; line number 379, marked in yellow.  

-page 13., subsection 4.4, the first paragraph; line number 553, marked in yellow.