Use of Eucalyptus Charcoal Waste in the Formulation of Substrate for the Cultivation of Two Strains (LED 20/11 and LED 20/12) of Lentinula edodes

Round 1

Reviewer 1 Report

The manuscript titled ‘’Use of Eucalyptus Charcoal Waste in The Lentinula edodes Substrate Production‘’ faces with some serious flaws. The abstract is not complete. The introduction is not sufficient at all, and the background of the research has been avoided. And more important, the paper doesn’t seem to be fitted with the scope of Agronomy journal since agronomy factors are not considered in this paper such as i) field experiment ii) long-term measurements (it should be at least two-years study) iii) agronomy analysis.

There are just some comments;

Line 12: the abstract needs to be started with the background of the idea and your main concern by doing this study! Needs to be precisely rephrased!

Line 16: following doses: 1%, 2%, 4%, 8%, 16% -> on which base are these percentages? WW, VV?

Line 23: abstract has not been finalized! What conclusion or future aspect you can draw from it?

The conclusion is like repeating the results!

Author Response

Reviewer 1

The manuscript titled ‘’Use of Eucalyptus Charcoal Waste in The Lentinula edodes Substrate Production‘’ faces with some serious flaws. The abstract is not complete. The introduction is not sufficient at all, and the background of the research has been avoided. And more important, the paper doesn’t seem to be fitted with the scope of Agronomy journal since agronomy factors are not considered in this paper such as i) field experiment ii) long-term measurements (it should be at least two-years study) iii) agronomy analysis.

Dear reviewer, we greatly appreciate the thorough review and constructive suggestions provided for our work. We deeply value the time and expertise you have dedicated to evaluating our study, and we are confident that your recommendations have significantly improved the quality of the work. 

There are just some comments;

Line 12: the abstract needs to be started with the background of the idea and your main concern by doing this study! Needs to be precisely rephrased!

The considerations have been added to the abstract.

Line 16: following doses: 1%, 2%, 4%, 8%, 16% -> on which base are these percentages? WW, VV?

The dosages were consistently doubled from the previous dosage until reaching a quantity of 16% of the substrate. The percentage corresponds to its amount in the dry weight of substrate formulation.

Line 23: abstract has not been finalized! What conclusion or future aspect you can draw from it?

The necessary alterations were made to the abstract.

The conclusion is like repeating the results!

The necessary alterations were made.

Reviewer 2 Report

The MS deals with the examination of using Eucalyptus fine charcoal FC as a    substrate source for Lentinula edodes mushroom production.

 The FC is a byproduct of Eucalyptus waste burning.

The authors have examined the reuse of this waste, which is an important goal.

 a

While the purpose of this study is important, I find some limitations.

 My main concern is that the study used only 2 strains of Lentinus, showing different reactions of the strains to FC.  Hence it is difficult to reach a conclusion. It is more of a case study.

To obtain overall conclusions of the effect of the FC on Lentinus growth, more strains need to be studied.

Here are some detailed comments:

Introduction:

1.               What is the significance of Lentinus.  What is its importance regarding the aim of using FC.     Please add the advantage of using the FC with Lentinus rather than with other mushroom substrates.

3.               Line 45-     

“The best substrate to use in the production of shiitake is the one that has the lowest economic cost and provides greater yield, considering the qualitative aspects of mushrooms (number and size)

 “

This is a general sentence and is not unique to Lentinus. 

4.               Line 58-

“Viotto et 58 al. [14] reported that the use of FC due to assistance in moisture retention and oxygenation 59 of the L. edodes substrate.”  

Please add the result of this work. i.e. did the use of FC increase yield?

Methods and materials

1.             Table 1: What do you mean by substrate? Is it the percent of FC in the substrate? Please elaborate.

2.             Table 1: Are the data of the sawdust and the FC taken from another work (from 2013)? It is expected that this will be examined for fresh sawdust and FC batch used in the present work and then will be compared to the new substrate mixture using the different FC in this work.

3.             Table 1: K₂O- Why have you shown the concentration of this compound and not that of standard K?

4.             Please clarify why you have used only 2 strains of Lentinus? It is expected to use at least 3 strains or more.

Results

1.             Do Table 2 and Figures 1 and 2 show the results of the 0 FC treatment of the FC experiment? Please indicate in the figure if the results are from the FC experiment or another experiment.

2.             Please add standard deviation in the table and figures.

3.             Table 3: What do the data represent? What is the Lentinus strain? Alternatively, is it a combination of the data of the two strains? What kind of combination? Average? Sum? Please add this information to the table.

4.             Figure 3: Since the figure represents an XY plot, it will be preferable to use X axis as linear data of FC concentration and not as the number of treatments. You may get different conclusions.

Conclusion

This section is a summary of the results.

Conclusions need to be added. Does the addition of FC improve yield, or at least does not harm it, and so on? Do you recommend its use? What is required to be further studied in this regard?  

Author Response

Reviewer 2

The MS deals with the examination of using Eucalyptus fine charcoal FC as a    substrate source for Lentinula edodes mushroom production. The FC is a byproduct of Eucalyptus waste burning. The authors have examined the reuse of this waste, which is an important goal. While the purpose of this study is important, I find some limitations. My main concern is that the study used only 2 strains of Lentinus, showing different reactions of the strains to FC.  Hence it is difficult to reach a conclusion. It is more of a case study. To obtain overall conclusions of the effect of the FC on Lentinus growth, more strains need to be studied.

Dear reviewer, we greatly appreciate the thorough review and constructive suggestions provided for our work. We deeply value the time and expertise you have dedicated to evaluating our study, and we are confident that your recommendations have significantly improved the quality of the work.

Here are some detailed comments:

Introduction:

1. What is the significance of Lentinus.  What is its importance regarding the aim of using FC.     Please add the advantage of using the FC with Lentinus rather than with other mushroom substrates.

The considerations were added to the introduction.

2. Line 45- “The best substrate to use in the production of shiitake is the one that has the lowest economic cost and provides greater yield, considering the qualitative aspects of mushrooms (number and size) “ This is a general sentence and is not unique to Lentinus. 

The expression was corrected to encompass the entire mushroom production.

3. Line 58- “Viotto et 58 al. [14] reported that the use of FC due to assistance in moisture retention and oxygenation 59 of the  edodes substrate.”  Please add the result of this work. i.e. did the use of FC increase yield?

Due to the nature of the scope of this work, no analyses were conducted to determine yield parameters. It is noteworthy that the substrates used were provided by regional producers, who employ eucalyptus charcoal to optimize moisture retention and substrate oxygenation. This practice is widely employed in mushroom cultivation, particularly in the formation of the surface layer intended for mushroom production (Eira et al., 2021).

Eira et al. (2021) Revisiting the casing layer: Casing materials and management in Agaricus mushroom cultivation. https://doi.org/10.1590/1413-70542021450001R21

Methods and materials

1. Table 1: What do you mean by substrate? Is it the percent of FC in the substrate? Please elaborate.

The description was changed as requested.

2. Table 1: Are the data of the sawdust and the FC taken from another work (from 2013)? It is expected that this will be examined for fresh sawdust and FC batch used in the present work and then will be compared to the new substrate mixture using the different FC in this work.

Yes, the data regarding FC and sawdust are taken from other studies. However, the data serve only to exemplify the chemical conditions of the initial material, as the chemical analysis for each substrate was determined.

3. Table 1: K₂O- Why have you shown the concentration of this compound and not that of standard K?

It's common practice for us to present the data in the K₂0 format; however, they have been modified and are now being presented in standard K format.

4. Please clarify why you have used only 2 strains of Lentinus? It is expected to use at least 3 strains or more.

We used the strains that have been yielding the best results in our research, which were employed. Furthermore, the strains were isolated from different regions under distinct cultivation conditions, and it is expected that they would suffice to demonstrate the variability exerted by the genetics of the strains.

Results

1. Do Table 2 and Figures 1 and 2 show the results of the 0 FC treatment of the FC experiment? Please indicate in the figure if the results are from the FC experiment or another experiment.

The table 2 presents the arithmetic mean of the various dosages observed in each studied lineage. Figure 1, in turn, represents the experimental dataset, encompassing all treatments and repetitions conducted. Lastly, Figure 2 comprises a comprehensive analysis of the experiment, also considering the mean of each investigated production flow. Supplementary information has been included in the tables and figures to provide a deeper understanding of the data analyses conducted.

2. Please add standard deviation in the table and figures.

Standard deviation has been incorporated into the table data, while the figures have not received this inclusion due to representing the same dataset, resulting in the application of the same standard deviation to the data contained within them.

3. Table 3: What do the data represent? What is the Lentinus strain? Alternatively, is it a combination of the data of the two strains? What kind of combination? Average? Sum? Please add this information to the table.

The data in Table 3 represent the average of observations made in the two lineages used in the study. This information has been included in the table to facilitate the understanding of the results.

4. Figure 3: Since the figure represents an XY plot, it will be preferable to use X axis as linear data of FC concentration and not as the number of treatments. You may get different conclusions.

 We acknowledge the observation made. However, the purpose of Figure 3 is to exemplify the variations in means concerning the applied doses, using the mean comparison test.

Conclusion

This section is a summary of the results.

Conclusions need to be added. Does the addition of FC improve yield, or at least does not harm it, and so on? Do you recommend its use? What is required to be further studied in this regard?  

The recommended changes have been implemented.

Round 2

Reviewer 1 Report

The paper has been improved and potentially can be published.

Author Response

Thank you very much for the time you dedicated to reviewing the work. Their contributions were fundamental to improving its quality and accuracy.

Reviewer 2 Report

The MS of the version has been improved.

Yet, the fact that the work is showing only 2 strains is limiting and can not give a general conclusion on the effect of the FC on the Shitake yield. 

With that, it is important to show the specific reaction of certain strains to the FC. 

comments

1. Please change the MS title to be more precise, i.e. "Use of Eucalyptus charcoal waste in the Lentinula edodes sub-strate production of XX and YY strains. " or a similar detailed title. 

2. Line 12-  Please add the scientific name (Lentinus edodes ) along with Shitake.

3. Table 3- What is the SD refer to?  In my former review, I suggested adding the SD of the results within each treatment, i.e. each FC concentration and each flush.  This will give an idea of the variation of yield within the repeats of each treatment. 

4. Line 240 – Use either shitake or Lentinus, but not both, in the same paragraph

Author Response

The MS of the version has been improved.

Yet, the fact that the work is showing only 2 strains is limiting and can not give a general conclusion on the effect of the FC on the Shitake yield. 

With that, it is important to show the specific reaction of certain strains to the FC. 

Dear Reviewer,

The use of 2 strains is precisely to demonstrate a duplicate genetic effect. No more strains were used, as these are used commercially in the country. It would not be agronomically viable to import a strain from another country for such a study. Thank you very much for the time you dedicated to reviewing the work. Your contributions will be fundamental to improving its quality and accuracy. Their considerations were accepted and implemented, changes were also made to the experimental design and presentation of results.

Comments

1. Please change the MS title to be more precise, i.e. "Use of Eucalyptus charcoal waste in the Lentinula edodes sub-strate production of XX and YY strains. " or a similar detailed title.

Dear reviewer, we agree with the change of title, it was changed to: “Use of Eucalyptus Charcoal Waste in the formulation of sub-strate for the cultivation of two strains (LED 20/11 and LED 20/12) of Lentinula edodes.”

2. Line 12- Please add the scientific name (Lentinus edodes) along with Shitake.

We made the correction, adding the scientific name in parentheses, along with the popular name. Due to the text changes, the change will now be found on line 13.

3. Table 3- What is the SD refer to?  In my former review, I suggested adding the SD of the results within each treatment, i.e. each FC concentration and each flush.  This will give an idea of the variation of yield within the repeats of each treatment.

 We added the standard deviation values in all treatments, shown in tables 2 and 3. The lower caption was also added, demonstrating what ± refers to.

4. Line 240 – Use either shitake or Lentinus, but not both, in the same paragraph

We check and make corrections, leaving only one name, whether popular or scientific.