Distinct Nutritional Profiles of Fermented Chamerion angustifolium from Natural, Organic, and Biodynamic Cultivation Systems: Evidence from a Four-Year Study

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript describes the 4-year study of nutritional profiles (vitamins, pigments, sugars) of fireweed leaves fermented by solid-state fermentation method. The impact of three cultivation systems of fireweed were compared (natural, organic, biodynamic) as well as the time of SSF (24 and 48 hours).

In my opinion the methods used in the research are suitable and results are described and discussed properly. However, I have a few comments.

First of all, the introduction is difficult to read due to its lack of consistency. It would be helpful to describe the biodynamic system (lines 45-46). The rest of the introduction requires careful review because it is difficult to read (lines 47-78). First, SSF is described, then there is a reference to fermented tea, followed by another mention of SSF, and then the description of fermented tea continues. After that, the topic of SSF again continues. Therefore, this section requires some tidying up. There is a cut sentence in lines 53-54. Additionally, the Results and Discussion section (lines 93-104) presents the definition of SSF again, which I consider unnecessary.

When citing, for example, "Giannattasio et al." the publication date is given in parentheses (e.g., lines 156, 161, 165, 171, 173, 201, etc.). I haven't encountered this type of citation in other manuscripts published in this journal, so it probably requires correction. In the Materials and Methods section (line 377), the figure title is given (it should be removed). Line 390 states "Error! Reference not found" - this is also likely an oversight. Figure 3 is duplicated, and some text from section 3.4 appears between the images.

Author Response

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Author Response File: Author Response.doc

Reviewer 2 Report

Comments and Suggestions for Authors

Review

on the paper "Distinct Nutritional Profiles of Fermented Chamerion angustifolium from Natural, Organic, and Biodynamic Cultivation Systems: Evidence from a Four-Year Study"

by Marius Lasinskas, Elvyra Jarienė, Jūratė Staveckienė, and Jurgita Kulaitienė

 

Main remarks

1. The study was conducted over four years. However, this is not reflected in the results: there is no information on the differences in the content of the studied metabolites by year and on the weather conditions during these four seasons.
2. During solid-state fermentation, the content of some metabolites increases by 24 hours and decreases by 48 hours. Or, conversely, they initially decrease and then increase. Overall, this pattern of changes in metabolite content during solid-state fermentation is atypical and requires speculation about the causes of the observed processes.
3. The surface of the fireweed leaves used for solid-state fermentation was untreated and, therefore, contaminated with bacteria and fungi. Their development during fermentation is confirmed by a decrease in glucose content, the most easily digestible saccharide. Since the microbial composition was not studied, the results of solid-state fermentation cannot be reproduced.
4. The authors did not draw a conclusion about the feasibility of solid-phase fermentation conducting, although samples obtained from this biotechnological process constituted the majority of samples used in the biochemical experiments.
5. There is no information on whether changes in leaf weight were observed after solid-state fermentation. Such changes in leaf mass could have affected metabolite content.

 

Minor remarks.

1. Table 3 is given twice.
2. The phrase in parentheses is unclear (L 390).

Author Response

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Author Response File: Author Response.doc

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript contains valuable experimental data and provides a useful comparison of cultivation systems and fermentation processes. However, several aspects could be improved to strengthen the scientific clarity, methodological transparency, and interpretation of the results.

Major comments

1. Novelty and scientific contribution

The study provides extensive compositional data; however, the novelty should be more clearly defined in the Introduction. The authors should better emphasize how their work advances current knowledge on fermentation of medicinal plants or on the nutritional enhancement of Chamerion angustifolium. A clearer comparison with previously published studies would help highlight the specific contribution of this work.

2. Experimental design clarification

The manuscript mentions that the study was conducted over four years, but the structure of the experimental design is not sufficiently clear. The authors should specify whether the data presented represent pooled results or averages across years, the number of biological replicates per treatment, how seasonal or interannual variability was handled statistically.

Clarifying these points would increase the reliability of the conclusions.

3. Statistical analysis and data presentation

Although ANOVA and Tukey tests are mentioned, the manuscript would benefit from a more detailed description of the statistical model used (factors included, interaction terms, etc.). Additionally, standard deviations or error bars should be consistently presented in tables or figures to indicate data variability.

4. Interpretation of fermentation effects

The discussion of biochemical mechanisms behind the observed changes in pigments, sugars, and vitamin C during fermentation could be expanded. For example, the potential role of microbial activity, enzymatic oxidation, or degradation pathways should be discussed more critically and supported with relevant literature.

5. Correlation analysis interpretation

The Pearson correlation tables are extensive, but their interpretation in the text remains limited. The authors should focus on the most biologically meaningful correlations and explain their relevance rather than listing many coefficients.

Minor comments

1. The manuscript would benefit from careful language editing to improve readability and reduce occasional grammatical inconsistencies.
2. Some sections of the Results and Discussion contain repetitive descriptions of data trends. Condensing these sections would improve the flow of the manuscript.
3. Figure captions should be expanded slightly to make the figures understandable without referring extensively to the main text.
4. Ensure consistent formatting of scientific names (e.g., Chamerion angustifolium should be italicized throughout the manuscript).
5. Some references appear somewhat dated; incorporating several recent studies (last 5 years) on plant fermentation and functional foods would strengthen the discussion.

Comments on the Quality of English Language

Long sentences can be written more clearly.

Author Response

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Author Response File: Author Response.doc

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors,

Thank you for your kind attention to my comments.

I would like to draw your attention to two issues.

1. I cannot agree with your response to comment 2. The manuscript does not explain why changes in the lutein, sucrose, glucose, and fructose contents in some experimental variants are atypical for fermentation processes, i.e., do not exhibit a consistent trend of these changes.
2. The response to comment 4 cannot be accepted. The possibility of conducting biochemical experiments cannot justify the development of any biotechnological process. The question was about what practical problems, or the increase in the content of which biologically active or nutrient substances, would benefit from solid-phase fermentation of fireweed leaves.

Comments for author File: Comments.docx

Author Response

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Author Response File: Author Response.doc

Reviewer 3 Report

Comments and Suggestions for Authors

What are the differences in nutritional profiles of the Fermented Chamerrion angustifolium from Natural, Organic, and Biodynamic Cultivation Systems, compared to only the biodynamic processes? The authors then compare natural food production processes and fermentation, and look to understand the chemistry of food fermentation in fireweed plants. It is of real interest from the point of view of sustainability and functional foods. So this is where the study provides valuable experimental results but, on the whole, the research provides a thorough look at how cultivation systems and fermentation are compatible in the study of food production. Major comments. 1. Novelty and scientific contribution. Although this study has a considerable composition and is highly diverse, the novelty deserves to be clearly described in an introductory paper; in that regard. This work will certainly contribute to the knowledge of fermentation of medicinal flora and is therefore important to the nutritional enhancement of Chamerion angustifolium. A clear picture of the specific contribution of this work would help compare with all previous studies considered in this research. 2. Experimental design clarification. Four years of experimental study was performed however the experimental format structure is not apparent for this study. The authors should ask why we are showing results. o the number of biological replicates per treatment (carotenoids, chlorophylls, sugars, and vitamin C derivatives) o this seasonal/interannual variation and when did we use them effectively? Let us clarify those points so that the conclusions will be more consistent. Statistical analysis and data presentation. Although ANOVA and Tukey tests have been used in the review, a more detailed description of the statistical model would help us understand why these findings occurred -factors used, interaction terms, etc. -must be given in the tables and in all figures as standard deviation or error bars so that the readers can see that it wasn’t just statistical noise and results in their statistics. The fermentation effects can also be interpreted as biochemical changes with the change of pigments or sugars (and Vitamin C) in fermentation). The authors should, for instance, address the possible role of microbial activity and enzymatic oxidation and degradation mechanisms separately and in line with the relevant research papers. 4. Correlation analysis interpretation The Pearson correlation tables are quite extensive, but in the text are relatively poorly summarised. It is critical. So, most correlations should be studied with the best likelihood to be biologically relevant, and in context of which the correlations were relevant rather than the list of coefficients. Some minor comments to begin with for the manuscript (language editing and the editing process). In the end there are some words for discussion, but a few sentences still need the edit in it. In the Research Section, results and discussion are repetitive in their comments. It would also contribute to chapter length if these sections were divided into smaller parts of the manuscript. Figure captions must be expanded a little with descriptive details about their structure that can be easily recognized using the figures but the text of the text itself should be not much larger to the point of loss of vocabulary. As well as, keeping scientific names and text formats appropriate in the manuscript. Name them properly and clearly (for example, Chamerion angustifolium should be italicized throughout the whole manuscript). 5. Some references look slightly dated; possibly several recent papers (last 5 years) on plant fermentation and functional foods would help us to keep up the discussion in advance of that. That’s it, the material has relevance in the work, and they have also given a good database. The manuscript had some deficiencies, but it provided insights into these deficiencies through experiments and added valuable evidence that can help us establish our work on functional plant products and fermentation-based processing.

Comments on the Quality of English Language

Long sentences can be written more clearly.

Author Response

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Author Response File: Author Response.doc

Round 3

Reviewer 3 Report

Comments and Suggestions for Authors

Manuscrpt was improved