Balanced Use of Zn, Cu, Fe, and B Improves the Yield and Sucrose Contents of Sugarcane Juice Cultivated in Sandy Clay Loam Soil
Round 1
Reviewer 1 Report
Dear Authors,
The manuscript entitled for review: "Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil" requires many important corrections. I have included all my comments in the pdf file as the reviewer comments.
Row 2. Juice quality is usually described by many parameters. In the paper presented for review, only polarization was marked. In my opinion, the title should be: Balanced use of micro-and macro-nutrients improves the yield and juice polarization of sugarcane cultivated in sandy loam soil
Row 24 Why was the @ symbol used?
Is it a dose of a micronutrient in a pure ingredient or in the form of ZnSO4 salt? The same question applies to the rest of the micronutrients
Row 87 US Salinity Laboratory Staff. Diagnosis and improvement of saline and alkali soils. Agriculture Handbook 60 Washington DC, United States Salinity Laboratory, USDA, 1954, p 160. Almost 70 years ago, modern analytical solutions were unknown. MDPI requires the use of modern research equipment. The authors refer to old methodologies that were once used. In the Materials and methods section, you can specify the source of the method, but you also need to describe the equipment that was used for testing. For example, EC was determined, the method was described in the cited literature, but a modern conductivity meter (Type, Manufacturer, City, Country) was used. The principle of descriptions of materials and methods should ensure the possibility of repeating the research elsewhere, by other authors, using the same research techniques!
This section needs very serious improvements!
Row 91 How do you know? Please enter particles size distribution and determination method! E.g. , the particle size distribution was determined using a Mastersizer 3000 (Malvern Instruments Limited, Worcestershire, UK)
or
Soil pH (pH) was determined potentiometrically in water and 1MKCl-1 solution in the ratio of 1: 2.5 (w / v), using a pH SenTix61 electrode and pH 538 WTW potentiometer (Xylem Analytics, Weilheim, Germany).
Row 93 No description for micronutrients! Why is the Zn content not specified? After all, it was one of the research factors!
Row 97 No methods described OM determination!
Row 106 The doses of fertilizers should be converted into the pure component: N, P, K
The same goes for micronutrients.
Of course, it is necessary to specify the form (fertilizers) in which these ingredients were applied.
Row 108 Common names should be avoided, here indicate potassium chloride or KCl or muriate of potash (KCl)
Row 114 Early maturity? Full maturity? When, please describe how was the harvest date determined?
As specified, stripped cane height (after all, the top part is cut off!)
What instrument was used to determine the Brix extract content?
Row116 There is no such % Brix unit.
Which refractometer was used to determine the sucrose content?
The Brix oBx scale can be used from which you can approximate % sucrose (but for sucrose content you need to use polarimeter!)
Of course in the Brix scale 1 oBx (Brix degree) approximately means 1 g of sucrose in 100 g of sugar/water solution.
Row 134 Did the authors also treat the year of research as a research factor?
Row 188-192 Authors described only one parameter, not parameters! You should explain what is Pol.
In figure 3, there is on Y-axis description Bx(%) should be Polarization or oBx, not %
Remember! Sucrose content% is not the same as polarization!
You can approximate the sugar content of the juice from the Bx readings, but it will not be pure sugar!
The quality parameters include:
The moisture content of cane
Brix of cane (Total dissolved Solids)
Polarization (Pol.) Of cane
Purity of cane
Reducing sugars of cane juice
the pH of cane juice
ash content
and macro and micronutrients content
How did the authors get the sugar yield? Is it a biological or technological sugar yield?
Row 203 In table 3 what means acronyms PKR and BCR? My guess is that PKR is the Pakistani currency. Can these values be converted into, for example, the more common USD?
And BCR is benefit-cost-ratio?
Row 213 Did the authors measure sucrose synthase activity? Are they authorized to make such statements? They can write that the use of Zn caused an increase in sucrose content, which may be related to an increase in sucrose synthase activity as reported by Pawar et al. [28]
The discussion of the results is very poorly developed!
Please note other minor corrections in the attached pdf file
Yours sincerely
Reviewer
Comments for author File: Comments.pdf
Author Response
Dear Authors,
The manuscript entitled for review: "Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil" requires many important corrections. I have included all my comments in the pdf file as the reviewer comments.
Response: We have incorporated all the comments/corrections suggested by the reviewers
Row 2. Juice quality is usually described by many parameters. In the paper presented for review, only polarization was marked. In my opinion, the title should be: Balanced use of micro-and macro-nutrients improves the yield and juice polarization of sugarcane cultivated in sandy loam soil
Response: As per the suggestion of the reviewer, we have revised the title as “Balanced use of Zn, Cu, Fe, and B improves the yield and sucrose contents of sugarcane juice cultivated in sandy clay loam soil”
Row 24 Why was the @ symbol used? Is it a dose of a micronutrient in a pure ingredient or in the form of ZnSO4 salt? The same question applies to the rest of the micronutrients
Response: It is a dose of micronutrients in the form of salt. We have replaced it with “at the rate of” and highlighted where changed
Row 87 US Salinity Laboratory Staff. Diagnosis and improvement of saline and alkali soils. Agriculture Handbook 60 Washington DC, United States Salinity Laboratory, USDA, 1954, p 160. Almost 70 years ago, modern analytical solutions were unknown. MDPI requires the use of modern research equipment. The authors refer to old methodologies that were once used. In the Materials and methods section, you can specify the source of the method, but you also need to describe the equipment that was used for testing. For example, EC was determined, the method was described in the cited literature, but a modern conductivity meter (Type, Manufacturer, City, Country) was used. The principle of descriptions of materials and methods should ensure the possibility of repeating the research elsewhere, by other authors, using the same research techniques! This section needs very serious improvements!
Response: We have revised the suggested section (Please see lines 88-93)
Row 91 How do you know? Please enter particles size distribution and determination method! E.g. , the particle size distribution was determined using a Mastersizer 3000 (Malvern Instruments Limited, Worcestershire, UK) or Soil pH (pH) was determined potentiometrically in water and 1MKCl-1 solution in the ratio of 1: 2.5 (w / v), using a pH SenTix61 electrode and pH 538 WTW potentiometer (Xylem Analytics, Weilheim, Germany).
Response: We have added the particle distribution in Table 1 and revised the suggested sentences as per the suggestion of the reviewer (Please see lines 96-97)
Row 93 No description for micronutrients! Why is the Zn content not specified? After all, it was one of the research factors!
Response: Zinc contents were medium in range; we have mentioned it in line 98
ECE?
Response: ECe means, electrical conductivity of extract
is it total Boron?
Response: No, it is available. Modified
Row 97 No methods described OM determination!
Response: Added as per the suggestion of the reviewer (Please see lines 95-96)
Row 106 The doses of fertilizers should be converted into the pure component: N, P, K. The same goes for micronutrients. Of course, it is necessary to specify the form (fertilizers) in which these ingredients were applied.
Response: Amended/revised and the form of nutrients/fertilizer is mentioned in treatments (Please see lines 116)
diammonium phosphate (NH4)2HPO4 is a source of both phosphorus and nitrogen! KCl source of potassium, why the authors write P and K sources respectively?
Response: We agree with the reviewer that diammonium phosphate (NH4)2HPO4 is a source of both phosphorus and nitrogen. We have modified the sentence to clarify the meaning (Please see lines 116-121)
Row 108 Common names should be avoided, here indicate potassium chloride or KCl or muriate of potash (KCl)
Response: Modified as per the suggestion of the reviewer (Please see line 118)
Row 114 Early maturity? Full maturity? When, please describe how was the harvest date determined? As specified, stripped cane height (after all, the top part is cut off!) What instrument was used to determine the Brix extract content?
Response: After twelve months the crop was harvested, sucrose of sugarcane juice was measured by Polarimeter (Please see line 129)
Row116 There is no such % Brix unit. Which refractometer was used to determine the sucrose content? The Brix oBx scale can be used from which you can approximate % sucrose (but for sucrose content you need to use polarimeter!). Of course in the Brix scale 1 oBx (Brix degree) approximately means 1 g of sucrose in 100 g of sugar/water solution.
Response: We agree with the reviewer. Polarimeter was used for measurement of pol reading to calculate sucrose contents in sugarcane juice sample (Please see line 131-132)
Row 134 Did the authors also treat the year of research as a research factor?
Response: Thanks for your comment. No, the 1st year data is of plant crop and 2nd year data is of ratoon crop
growth chemicals??
Response: Revised (Please see line 160)
Row 188-192 Authors described only one parameter, not parameters! You should explain what is Pol.
Response: Revised and Pol mean polarization (angle of rotation of sucrose) which was measured by using Polarimeter (Please see line 209).
In figure 3, there is on Y-axis description Bx(%) should be Polarization or oBx, not %
Remember! Sucrose content% is not the same as polarization!
You can approximate the sugar content of the juice from the Bx readings, but it will not be pure sugar!
The quality parameters include: The moisture content of cane, Brix of cane (Total dissolved Solids), Polarization (Pol.) Of cane, Purity of cane, Reducing sugars of cane juice, the pH of cane juice, ash content and macro and micronutrients content
Response: As per the suggestion of both the reviewers, we have mentioned POL% as measured through Polarimeter for the estimation of sucrose contents in sugarcane juice
How did the authors get the sugar yield? Is it a biological or technological sugar yield?
Response: Cane yield was multiplied with sugar recovery (%) of juice and divided by 100 to calculate sugar yield t/ha (Please see line 133).
??? what is RD
Response: RD means Recommended, revised, and highlighted (Please see Table 3)
Row 203 In table 3 what means acronyms PKR and BCR? My guess is that PKR is the Pakistani currency and BCR is benefit-cost-ratio? Can these values be converted into, for example, the more common USD?
Response: Yes, PKR is a Pakistani Rupee, and BCR is a benefit-cost ratio. We have converted PKR to USD as per the suggestion of the reviewer (Please see Table 3)
Row 213 Did the authors measure sucrose synthase activity? Are they authorized to make such statements? They can write that the use of Zn caused an increase in sucrose content, which may be related to an increase in sucrose synthase activity as reported by Pawar et al. [28]
Response: Revised as suggested (Please see lines 235-236)
The discussion of the results is very poorly developed!
Response: We have revised the discussion section as per the suggestion of the reviewer and highlighted it as yellow where changes were made
Please note other minor corrections in the attached pdf file
Response: We have incorporated/corrected all the suggestions/comments and highlighted
Yours sincerely
Reviewer
Author Response File: Author Response.docx
Reviewer 2 Report
The manuscript contains interesting experimental results. Their value is lowered by the fact that the results come from 2 years and the number of repetitions of each combination is 3, not 4. The Materials and Methods chapter needs to be corrected. The discussion is not extensive and requires supplementing and refinement.
Comments:
The title is in the form of an application. I propose to change it.
Citation: Lastname, F.; Lastname, F.; Lastname, F. Title. - is not completed.
Line 69, [5,12]. The order should not be 12, but 8
Line 81, please include a figure with the location of the experiment
Line 88, pH in KCl or pH in H2O?
Line 89, what was the available magnesium content in the soil?
Line 93, please provide the soil classification according to to IUSS Working Group WRB. World Reference Base for Soil Resources 2014. International Soil Classification System for Naming Soils and Creating Legends for Soil Maps. Update 2015;World Soil Resources Raport 106; FAO: Rome, Italy, 2015; 188 p.
Table 1, instead of organic matter, please provide the content of soil organic carbon (SOC)
Line 102, etc., correct 0.2 kg ha-1
Line 102, what is borax? Please provide the name of the compound or the chemical formula
Line 106, please provide the doses of phosphorus and potassium in kg P and kg K ha-1
Line 107, please specify the composition of the mineral fertilizers used
Line 108, what nitrogen fertilizer was used? Please provide its composition
Figure 1, 2, a bit strange form of figure
Table 3, there is no statistical analysis
Table 3, please explain what is PKR? Please enter in USD.
References, more than half of the publications are over 10 years old - before 2010 (4, 13-19, 21-26, 28-30, 32, 33). Please remove them. It is necessary to supplement with the latest publications.
Author Response
The manuscript contains interesting experimental results. Their value is lowered by the fact that the results come from 2 years and the number of repetitions of each combination is 3, not 4. The Materials and Methods chapter needs to be corrected. The discussion is not extensive and requires supplementing and refinement.
Response: The 1st year data is of plant crop and 2nd year data is of ratoon crop. Revised and improved as suggested.
Comments:
The title is in the form of an application. I propose to change it.
Response: We have revised the title as “Balanced use of Zn, Cu, Fe, and B improves the yield and sucrose contents of sugarcane juice cultivated in sandy clay loam soil”
Citation: Lastname, F.; Lastname, F.; Lastname, F. Title. - is not completed.
Response: It will be completed once the paper has been accepted
Line 69, [5,12]. The order should not be 12, but 8
Response: We have crosschecked the order and found it correct as we already listed 11 references citations
Line 81, please include a figure with the location of the experiment
Response: We have added the GPS points of the study area location (Please see lines 82-83)
Line 88, pH in KCl or pH in H2O?
Response: Modified as per the suggestion of the reviewer “The pH was measured from 1:2.5 soil water ratio, respectively while the extract from the soil saturated paste was used for EC determination” (Please see lines 92-93)
Line 89, what was the available magnesium content in the soil?
Response: Unfortunately, we have not determined magnesium as it is not deficient in Pakistani soils
Line 93, please provide the soil classification according to IUSS Working Group WRB. World Reference Base for Soil Resources 2014. International Soil Classification System for Naming Soils and Creating Legends for Soil Maps. Update 2015; World Soil Resources Report 106; FAO: Rome, Italy, 2015; 188 p.
Response: We have added the soil classification as per the USDA soil taxonomy and WRB soil classification systems (Please see lines 96-97)
Table 1, instead of organic matter, please provide the content of soil organic carbon (SOC)
Response: We have added the contents of soil organic carbon (SOC) instead of organic matter (Please see Table 1)
Line 102, etc., correct 0.2 kg ha-1
Response: Corrected (Please see line 105)
Line 102, what is borax? Please provide the name of the compound or the chemical formula
Response: We have added the chemical formula of borax (Na2B4O7·10H2O) used in the present study
Line 106, please provide the doses of phosphorus and potassium in kg P and kg K ha-1
Response: We have added the recommended NPK doses (Please see line 116)
Line 107, please specify the composition of the mineral fertilizers used
Response: We have added the composition of all the fertilizers used in the present study (Please see lines 117-118)
Line 108, what nitrogen fertilizer was used? Please provide its composition
Response: Urea and diammonium phosphate were used as N sources. We have added the composition of both fertilizers (Please see lines 117-118)
Table 3, there is no statistical analysis
Response: In Table 3, the data presented is easily understandable where there is no need for statistical analysis
Table 3, please explain what is PKR? Please enter in USD.
Response: We have converted the Pakistani rupee to USD as per the suggestion of the reviewer
References, more than half of the publications are over 10 years old - before 2010 (4, 13-19, 21-26, 28-30, 32, 33). Please remove them. It is necessary to supplement with the latest publications.
Response: New references have been added
Author Response File: Author Response.docx
Reviewer 3 Report
Review of the article: “ Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil”
I suggest changing the title - the research involved differentiated fertilization only with microelements: Zn, Cu, Fe, B
The macronutrient fertilization was the same in all combinations. Maybe the title would be better
“Balanced use of Zn, Cu, Fe and B improves the yield and juice quality of sugarcane cultivated in sandy loam soil”
Modify the aim of research L 75-76 accordingly
- Materials and Methods
L 92 and Table 1. Please refer to the standards used to describe the soil nutrient levels - low, medium high. Quote relevant literature.
Were the plants grown in the same soil in both years - was the fertilization of the plants the same in each year of the study. Every year plants were planted or the same plants were grown for two tears on the same filed. This must be described in detail. Not every reader knows sugarcane cultivation agrotechnics - I, as a reviewer, do not know it and please describe the cultivation method in more detail
Which year are the results in table 1 - before and after cultivation?
There is no table with meteorological data during the research - plant cultivation. Precipitation, insolation, temperature - please complete if possible.
L 99-105 and the remaining manuscript fragments
Doses: 02 or 2.0 or maybe 20.0? Do not write 05 and 01 or 02 only 2.0, 1.0 5.0 etc.
On what basis were the doses of micronutrients determined - cite the literature or describe the preliminary research.
L 99-105 and description of all figures ”Replace the“ @ ”symbol with a description way.
Description of all figures; complete what do the letters of homogeneous groups mean? What do the bars indicate?
L 189. What does the abbreviation "Pol ..." mean. Brx is a refractometric measurement and not a polarimetric (compare with L 113-117). This is incomprehensible. In the entire publication I did not find an explanation for the abbreviation "Pol."
L 113-117 The description of the measurement methods for sugars and brix should be more precise. This fragment of the analysis method requires a detailed description
Table 3. Below this table will explain all abbreviations used in this table. Specify the units used in this table.
Since the research covered only four micronutrients, please describe in the final part of the discussion the potential influence of other micronutrients on metabolism - the sugar content in plants, e.g. molybdenium, manganeseiodine, selenium, vanadium, etc. Please try to write a short comment about the potential influence of other elements on the sugar content. If necessary, describe the need for study with other elements. See for example:
Sugar Tech 2018, 20, 518–533; or Front. in Plant Sci. doi: 10.3389/fpls.2021.656283; or
Agronomy https://doi.org/10.3390/agronomy11091720
or use other literature.
Author Response
Review of the article: “Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil” I suggest changing the title - the research involved differentiated fertilization only with microelements: Zn, Cu, Fe, B. The macronutrient fertilization was the same in all combinations. Maybe the title would be better. “Balanced use of Zn, Cu, Fe, and B improves the yield and juice quality of sugarcane cultivated in sandy loam soil”
Response: We have modified the title as “Balanced use of Zn, Cu, Fe, and B improves the yield and sucrose contents of sugarcane juice cultivated in sandy clay loam soil”
Modify the aim of research L 75-76 accordingly
Response: We have modified the aim of the research (Please see lines 78-80)
Materials and Methods
L 92 and Table 1. Please refer to the standards used to describe the soil nutrient levels - low, medium high. Quote relevant literature.
Response: We have added the relevant literature as per the suggestion of the reviewer (Please see lines 110-111)
Were the plants grown in the same soil in both years - was the fertilization of the plants the same in each year of the study. Every year plants were planted or the same plants were grown for two tears on the same field. This must be described in detail. Not every reader knows sugarcane cultivation agrotechnics - I, as a reviewer, do not know it and please describe the cultivation method in more detail
Response: We have added the suggested details (Please see lines 112-115)
Which year are the results in table 1 - before and after cultivation?
Response: Before the start of the experiment and at the end of the experiment in November 2021
There is no table with meteorological data during the research - plant cultivation. Precipitation, insolation, temperature - please complete if possible.
Response: We have meteorological data during the study period (Figure 1)
L 99-105 and the remaining manuscript fragments Doses: 02 or 2.0 or maybe 20.0? Do not write 05 and 01 or 02 only 2.0, 1.0 5.0 etc.
Response: Revised
On what basis were the doses of micronutrients determined - cite the literature or describe the preliminary research.
Response: We have cited the literature from which the doses were selected (Please see lines 111)
L 99-105 and description of all figures ”Replace the“ @ ”symbol with a description way.
Response: We have revised the description of all Figures (Please see the highlighted text)
Description of all figures; complete what do the letters of homogeneous groups mean? What do the bars indicate?
Response: Revised as per the suggestion of the reviewer (Please see the highlighted text)
L 189. What does the abbreviation "Pol ..." mean. Brx is a refractometric measurement and not a polarimetric (compare with L 113-117). This is incomprehensible. In the entire publication, I did not find an explanation for the abbreviation "Pol."
Response: We have explained “Pol”. Also revised the materials and methods section and Figure 3 accordingly
L 113-117 The description of the measurement methods for sugars and brix should be more precise. This fragment of the analysis method requires a detailed description
Response: Revised as per the suggestion of the reviewer
Table 3. Below this table will explain all abbreviations used in this table. Specify the units used in this table.
Response: Revised
Since the research covered only four micronutrients, please describe in the final part of the discussion the potential influence of other micronutrients on metabolism - the sugar content in plants, e.g. molybdenum, manganese, iodine, selenium, vanadium, etc. Please try to write a short comment about the potential influence of other elements on the sugar content. If necessary, describe the need for study with other elements. See for example:
Sugar Tech 2018, 20, 518–533; or Front. in Plant Sci. doi: 10.3389/fpls.2021.656283; or
Agronomy https://doi.org/10.3390/agronomy11091720
or use other literature.
Response: We have consulted the suggested papers, cited and added the suggested statement (Please see lines 251-253)
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Dear Authors,
The manuscript entitled for review: "Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil" still requires minor corrections. I have included all my comments in the pdf file as the reviewer comments.
Row 22. Please change the sentence as below: The aim of this study was to investigate the effect of micronutrient application on the yield and sucrose content expressed as the polarization of sugar cane juice (POL%) under field conditions.
Row 24 Why was the @ symbol used?
Is it a dose of a micronutrient in a pure ingredient or in the form of ZnSO4 salt? The same question applies to the rest of the micronutrients
Row 91 Please type (Type, Manufacturer, City, Country) of apparatus was used.
Row 95 Please provide a method of Zn determination as I suggested previously!
Row 101 Please enter particles size distribution and determination method!
E.g. , the particle size distribution was determined using a Mastersizer 3000 (Malvern Instruments Limited, Worcestershire, UK) or hydrostatic method?
How did you convert Organic matter % (see first version of manuscript) into SOC % (current version)?
0.48 -> 0.278
and
0.51 -> 0.296 ?
The discussion of the results is still very poorly developed!
Please note other minor corrections in the attached pdf file
Yours sincerely
Reviewer
Comments for author File: Comments.pdf
Author Response
Response to Reviewer comments
Dear Authors,
Comment: The manuscript entitled for review: "Balanced use of micro-and macro-nutrients improves the yield and juice quality of sugarcane cultivated in sandy loam soil" still requires minor corrections. I have included all my comments in the pdf file as the reviewer comments.
Response: Thanks for your comments. We have incorporated all the comments/suggestions made and highlighted as yellow where changes were made
Comment: Row 22. Please change the sentence as below: The aim of this study was to investigate the effect of micronutrient application on the yield and sucrose content expressed as the polarization of sugar cane juice (POL%) under field conditions.
Response: We have changed the suggested sentence (Please see lines 22-24)
Comment: Row 24 Why was the @ symbol used? Is it a dose of a micronutrient in a pure ingredient or in the form of ZnSO4 salt? The same question applies to the rest of the micronutrients
Response: We applied different micronutrients doses as their salts. We have modified the details of the treatments and highlighted where revised
Comment: Row 91 Please type (Type, Manufacturer, City, Country) of apparatus was used.
Response: Added (Please see lines 92-93)
Comment: Row 95 Please provide a method of Zn determination as I suggested previously!
Response: We have added the micronutrients (Zn, Cu, Fe, and Mn) determination method (Please see lines 97-98)
Comment: Row 101 Please enter particles size distribution and determination method! E.g. , the particle size distribution was determined using a Mastersizer 3000 (Malvern Instruments Limited, Worcestershire, UK) or hydrostatic method?
Response: We have added the particles size distribution and determination method as “The particle size distribution for soil textural analysis was determined by the hydrometer method as described by Bouyoucos [13] and Page [14].” (Please see lines 88-90)
Comment: How did you convert Organic matter % (see first version of manuscript) into SOC % (current version)? 0.48 -> 0.278 and 0.51 -> 0.296 ?
Response: As we presented the value of organic matter earlier. It was converted into organic carbon via the following formula
Organic matter (%) = 1.724 * total organic carbon
Comment: The doses of fertilizers should be converted into the pure components: N, P, K. The same goes for micronutrients. Of course, it is necessary to specify the form (fertilizers) in which these ingredients were applied
Response: We have converted the fertilizer doses as “pure NPK 168:48.9:92.6 kg ha-1” (Please see lines 117-118)
Comment: The methodology should describe exactly that 1st year data is of plant crop and 2nd year data is of ratoon crop. Write what is the difference! The reader from the US or Europe is not familiar with the technology of growing sugar cane, so it should be explained!
Response: We have added a statement as per the suggestion of the reviewer (Please see lines 131-132)
Comment: The discussion of the results is still very poorly developed!
Response: We have modified/revised the discussion to make it deep and well developed and highlighted as yellow where changes were made
Author Response File: Author Response.docx