Nanoformulations of the Piper auritum Kunth (Piperales: Piperaceae) Essential Oil for the Control of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

Round 1

Reviewer 1 Report (Previous Reviewer 2)

Comments and Suggestions for Authors

I have carefully compared and revised the new version of the resubmitted  manuscript entitled “Nanoformulations of the Piper auritum Kunth (Piperales: Piperaceae)essential oil for the control of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)”(agriculture-4114243-v1 versus agriculture-4114243 - with tack changes.
The revised manuscript presents the development and evaluation of three nano-based formulations of Piper auritum essential oil: a nanoemulsion (NE), a microemulsion (ME), and silver nanoparticles (AgNP), as potential eco-friendly tools for controlling Spodoptera frugiperda. The work remains novel in its combined approach: green synthesis of silver nanoparticles using essential oil, comparison of NE, ME, and AgNP systems and biological evaluation against S. frugiperda under controlled conditions.

Compared to the original version (agriculture-4114243-v1), the revised manuscript now includes additional stability studies of essential oil formulations over 60 days, a clearer description of nanoparticle dispersion and polydispersity, expanded bioassay methodology, including replication and controls and improved discussion of variability in essential oil composition and environmental limitations. Among the tested formulations, the nanoemulsion showed the highest larvicidal activity is supported by morphological observations of larval and pupal damage.

The scientific quality of the manuscript has clearly improved compared to version “agriculture-4114243-v1”. The authors addressed major concerns related to:

1. Lines 195-225: Reproducibility of bioassays
2. Lines 246-255: Inclusion of controls and statistical treatment
3. Lines 181-193: Formulation stability

The experimental design is now transparent and reproducible.

The added discussion (Lines 568-594) on environmental impact and formulation stability strengthens the relevance to the application. The Conclusion is moderated and includes limitations. The supplementary material has been improved and logically linked to the manuscript.

Overall, all concerns raised in the resubmitted manuscript are adequately addressed.

Only minor issues remain:

1. A final English language polishing is recommended to improve flow and reduce sentence length.
2. Minor formatting consistency (units, spacing) should be checked during production.

Author Response

Comments and Suggestions for Authors

I have carefully compared and revised the new version of the resubmitted manuscript entitled “Nanoformulations of the Piper auritum Kunth (Piperales: Piperaceae) essential oil for the control of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)”(agriculture-4114243-v1 versus agriculture-4114243 - with tack changes.
The revised manuscript presents the development and evaluation of three nano-based formulations of Piper auritum essential oil: a nanoemulsion (NE), a microemulsion (ME), and silver nanoparticles (AgNP), as potential eco-friendly tools for controlling Spodoptera frugiperda. The work remains novel in its combined approach: green synthesis of silver nanoparticles using essential oil, comparison of NE, ME, and AgNP systems and biological evaluation against S. frugiperda under controlled conditions.

Compared to the original version (agriculture-4114243-v1), the revised manuscript now includes additional stability studies of essential oil formulations over 60 days, a clearer description of nanoparticle dispersion and polydispersity, expanded bioassay methodology, including replication and controls and improved discussion of variability in essential oil composition and environmental limitations. Among the tested formulations, the nanoemulsion showed the highest larvicidal activity is supported by morphological observations of larval and pupal damage.

The scientific quality of the manuscript has clearly improved compared to version “agriculture-4114243-v1”. The authors addressed major concerns related to:

1. Lines 195-225: Reproducibility of bioassays
2. Lines 246-255: Inclusion of controls and statistical treatment
3. Lines 181-193: Formulation stability

The experimental design is now transparent and reproducible.

The added discussion (Lines 568-594) on environmental impact and formulation stability strengthens the relevance to the application. The Conclusion is moderated and includes limitations. The supplementary material has been improved and logically linked to the manuscript.

Overall, all concerns raised in the resubmitted manuscript are adequately addressed.

Only minor issues remain:

Comment 1: A final English language polishing is recommended to improve flow and reduce sentence length.

Response: The authors thank the reviewer for their comments. The document has been reviewed and corrected. Please see the text highlighted in yellow.

Comment 2: Minor formatting consistency (units, spacing) should be checked during production.

Response: The authors thank the reviewer for their comments. The document has been reviewed and corrected. Please see the text highlighted in yellow.

 

Author Response File: Author Response.pdf

Reviewer 2 Report (Previous Reviewer 1)

Comments and Suggestions for Authors

Authors have made a conscientious effort to address the concerns raised in an earlier peer review, and the revised manuscript represents a substantial improvement over the original version.  I can now recommend that the paper be published.  Please note that genus names (in this case Piper) must, like the full Latin binomial, always be italicized.  This should be corrected throughout the manuscript, especially in the Discussion section.

Author Response

Comments and Suggestions for Authors

Authors have made a conscientious effort to address the concerns raised in an earlier peer review, and the revised manuscript represents a substantial improvement over the original version. I can now recommend that the paper be published.  Please note that genus names (in this case Piper) must, like the full Latin binomial, always be italicized.  This should be corrected throughout the manuscript, especially in the Discussion section.

Response: The authors thank the reviewer for their comments. The document has been reviewed and corrected. Please see the text highlighted in yellow.

Author Response File: Author Response.pdf

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

see attached file

Comments for author File: Comments.pdf

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript “Nanoformulations of the Piper auritum essential oil for the control of Spodoptera frugiperda(J.E. Smith) (Lepidoptera: Noctuidae)“ investigates the green synthesis of silver nanoparticles using Piper auritum essential oil and evaluates their biological activity against Spodoptera frugiperda. The study combines nanoparticle characterisation with biological assays, aiming to emphasise eco-friendly alternatives to conventional pest control. The main strengths are the application of green chemistry principles, the integration of chemical and biological evaluations, and the inclusion of supplementary material (UV-Vis, FTIR, microscopy, and bioactivity data), which provides additional support for the findings.

The manuscript addresses a relevant topic, bridging nanotechnology, plant-derived bioactive compounds, and the control of agricultural pests. However, the review of prior work on plant-based nanoparticle synthesis and insecticidal activity is somewhat limited. The inclusion of recent studies would make the paper more comprehensive. While the experiments appear generally well-designed, several issues concerning reproducibility and methodological transparency require improvement. For example, the nanoparticle size distribution and stability are insufficiently discussed, and biological assays lack detailed descriptions of replicates, controls, and statistics. The discussion should be improved by expanding of consideration of limitations, such as potential variability in essential oil composition, scalability of the synthesis, and environmental/ecotoxicological implications. Without these, the conclusions seem too strong in relation to the presented evidence.

The cited literature includes relevant sources, but many references are older or regional. More recent publications on nanoparticle synthesis and bioactivity testing should be incorporated. Self-citations are not excessive, but the balance could be improved.

Specific comments

Manuscript

Lines 112–130: More details are needed on reproducibility (number of synthesis repetitions, batch-to-batch variation).

Lines 178–195: The FTIR spectrum (Figure S3) is presented, but the interpretation is limited. A clearer assignment of functional groups contributing to nanoparticle stabilization would strongly support the discussion.

Table 2: Should specify the statistical tests used for biological assays and clarify sample sizes.

Conclusion: Conclusions should be moderated to reflect limitations in toxicity data and environmental impact. Future research directions, such as standardization of essential oils, nanoparticle stability testing, and field trials, should be emphasized.

Supplementary material

Figure S2: The effect of EO/DW ratio and AgNO₃ volume is shown, but quantitative analysis (particle size, polydispersity) is missing.

Figure S4: The biological images are informative, but results should be supported by quantitative data on egg-laying reduction, with error bars and statistical analysis.

Table S1: The table is valuable but could be better linked to the main text.

 

 

Author Response

Please see the attachment

Author Response File: Author Response.pdf