Sustainable Biomass Valorization by Solid-State Fermentation with the Mutant Strain Trichoderma viride M5-2 of Forage Legumes to Improve Their Nutritional Composition as Animal Feed
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript presents a solid state fermentation of two legumes to improve the nutritional value for animal feed. It presents experimentation, FT-IR characterization and several DoE. As it is, the document needs to improve in the below areas before publication.
Major revisions
The manuscript needs major revision in the English writing, Grammar and how ideas are presented. It is a must to perform a revision in all the document. There is a template to prepare a manuscript for this journal. Authors should strictly abide by the format. Please ensure that the document has the same format, it’s not optional.
The design of Experiments (DoE) is confusing. The presentation of the results does not give significant information. A DoE is used to study a phenomenon which goal is to establish the best operation conditions based on a response variable. I see too many analysis, response variables, but there is not a proposal on the best experimental conditions. In the conclusion there wasn’t stated the best way to process the meals according to the DoE.
Other revisions
Line 62-64, Where are the reports that support this claim? Is there a review? A series of articles? Authors should state a reference to sustain this claim
Line 73-75, According to this line previous works have done a very similar job than this manuscript more than 10 years ago (the reference is from 2014), so what is really the novelty of the work?
Line 86, What does ICA mean? Authors must put the meaning of any acronym in the manuscript.
Materials and Methods, This section lack of references in the procedures. There are complete paragraphs without a single one of them. The authors must use references to other works in which they discuss how they decide to use those analysis.
Line 161-170, In the factorial design, so there are 3 response variables? Fiber composition, DM and pH? The authors should state why they choose these variables as the key point to define their experiment.
Line 164-165, What are the authors trying to say here? Please rewrite the idea in this sentence.
Line 166-169, Are there 2 factorial design? Why the authors choose to expand the time in this one? Why change the response variable? Where all the experiments done in the same flask or in different ones?
Table 1 and Table 2, Why is the ANOVA not shown in the manuscript? These tables present the information on the results and the P value, but the complete ANOVA for each DoE is not presented. Authors must include this information for replication purposes. Also, add information regarding the residues of the experiment and the model equation. What other information can the authors add? Are they contour plot or response surface in which areas are more suitable for this experimentation?
Table 3, Is pH a variable that important to be considered as a response variable in a DoE? Or is more of a descriptive variable on how the experiment works? Im curious if the authors plan to choose experimentation conditions by the pH measurements.
Table 4 and 5, Again the authors should show the results of the DoE and then the complete ANOVA results. Add information regarding the residues of the experiment and the model equation. Are they contour plot or response surface in which areas are more suitable for this experimentation?
Table 3, 4, 5, 6, These results are similar into an ANOVA for a single factor than a factorial design. Are authors trying to select the better conditions in which the pH, % DM, Physical indicators, Solubility (%) are obtained? This is an important question, what would be the best set of conditions to improve the nutritional value of the flour, and why are they suggesting this parameters?
In the discussion there is no comparison between the nutritional value of the flours and other animal feed. How the authors can state that this meals are efficient in animals feed? (Line 456) The manuscript lack the information of nutritional values on animal feed in Ecuador or Cuba?
Author Response
First, we'd like to thank you for your time and suggestions for improving the quality of our article, Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors
(1) The innovation and practical application value of the research need to be more prominently highlighted in the abstract. It is recommended to add one or two sentences at the end of the abstract to emphasize the study's potential contributions to sustainable agriculture or the animal feed industry.
(2) The research gap needs to be more clearly identified. Currently, the introduction mentions the potential of fungal fermentation but does not explicitly state why Trichoderma viride M5-2 was chosen and how it differs from existing studies.
(3) A justification for the experimental design needs to be supplemented. For example, why was a 120-hour fermentation time selected? Is there any pre-experimental evidence to support this choice?
(4) The statistical analysis methods section requires more detailed explanation. For instance, are the specific model of ANOVA and the conditions for hypothesis testing satisfied?
(5) The description of statistical differences in some results is not sufficiently clear. For example, do the changes in "CP" and "TP" in Table 4 have biological significance? Additional discussion is needed.
(6) The discussion could focus more on the innovative aspects of the research. For example, in what ways does T. viride M5-2 outperform previous studies?
(7) The limitations of the study need to be supplemented. For example, could the rise in pH during the fermentation process affect the long-term stability of enzymes? Has the feasibility of large-scale production been considered?
(8) Some conclusions require more literature support. For instance, in the fourth paragraph, it is mentioned that "lignin concentration was due to two possible causes," but no relevant literature is cited to substantiate this claim.
(9) The quality of figures must be improved.
Author Response
First, we'd like to thank you for your time and suggestions for improving the quality of our article, Please see the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsAll of the comments were addressed in this revision. New supplementary data was provided, and the overall discussion of the research was improved
Author Response
Dear reviewer, we are pleased that you were satisfied with the changes we made to our manuscript based on your suggestions. We are convinced that your invaluable help allowed us to create an improved and more understandable version. Once again, all the authors thank you for your time and valuable advice.
Reviewer 2 Report
Comments and Suggestions for Authorsthe paper can be accepted in the present form.
Author Response
Dear reviewer, we are pleased that you were satisfied with the changes we made to our manuscript based on your suggestions. We are convinced that your invaluable help allowed us to create an improved and more understandable version. Once again, all the authors thank you for your time and valuable advice.
