Effects of Cynanchum bungei Decne Addition on the Physicochemical Properties and Antioxidant Activity of Rice Wine

Round 1

Reviewer 1 Report (Previous Reviewer 1)

The revised manuscript has been improved accordingly to suggestions

Author Response

Thanks very much, I am glad the manuscript is recognized by the reviewer.

Reviewer 2 Report (Previous Reviewer 2)

Authors made some corrections according to the reviewer’s suggestion. 

Some of the experimental methods were very confusing. In brewing process, Cynanchum bungei Decne homogenate was added as discirbed in line 65-66, however, in “2.10 Measurement of growth and glucose uptake of yeast”, as shown in line 139-140, Cynanchum bungei Decne was extracted and added to the culture medium. Both of the ratios were set as 5%, 15%, and 25%. Why not use the same extraction method? It is very hard to draw accurate conclusions by using different material and methods. 

It is very hard to understand why authors did the experiments in section 3.7. It is obvious in the section 3.7 Cynanchum bungei Decne extracts affected the fermentation of yeast in the later phase, but what about the homogenate? The results in section 3.7 did not support the rice wine fermentation. Author should track the changes of yeast activity during rice wine fermentation. 

As the results showed, adding 25% of Cynanchum bungei Decne (CYW) resulted in similar alcoholic content with 15% addition (BYW), higher bioactive compounds, higher antioxidant activity and more volatile compounds, only with lower sensory evaluation score. It is hasty to draw the conclusion by choosing 15% as the best addition ratio for the rice wine production.

 

Minors:

1.    Line 54, What do author mean by“commercial mature”? 

2.    Line 162, “1.5 mL culture solution of one week's”, from which culture solution?

3.    Line 173, “mM”should me “mmol/L”; Line 178,“M”should be “mol/L”; “mmol/L”or“mg mL^-1 ”, please use the same format.

 

Language should be further improved. 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report (New Reviewer)

The manuscript discusses the utilization of Cynanchum bungei in the production of rice wine. While the study presents some interesting findings, there are areas where the manuscript could be improved. Here are some considerations:

1. The introduction section is too concise. It is unclear whether the authors aim to introduce a new source of starch for the rice wine process or enhance its content of bioactives. I suggest that the authors expand this section to provide more clarity and context.

2. Once the complete species name "Cynanchum bungei" is mentioned for the first time in the main text, it should be abbreviated as "C. bungei" for all subsequent mentions throughout the manuscript. The repetition of the species' name, especially in the introduction section, affects the readability of the text. I recommend revising the manuscript to improve this aspect.

3. Some methodologies are poorly described. While it may not be necessary to provide a comprehensive description for every method, certain essential information should be included regardless of referencing external sources. Please revise sections 2.4, 2.7 (including a description of the minor modifications mentioned), 2.8, and 2.12 (please describe the Coomassie bright blue method for protein quantification) to ensure the methodology is adequately explained.

4. Please separate numerical values from their respective units throughout the entire text to improve clarity and readability.

5. The statistical analysis requires further clarification. It would be helpful to understand why the authors chose ANOVA for data analysis. Did they evaluate the distribution of data using a normality test? If so, which test did they perform? This information is important for ensuring the accuracy of the data analysis.

6. On page 7, did the authors perform correlation analysis? If so, this analysis should be described in the Materials and Methods section. Additionally, if a correlation matrix was generated, it should be included either in the main text or as a supplementary file, with appropriate references in the manuscript.

7. On page 9, the authors claim an inhibition effect during the stable phase of yeast growth. However, it appears that there is no statistical significance. Furthermore, relying solely on fluorescence microscopy is not sufficient to establish the interference of C. bungei in cell viability. Figure 4 serves as an illustration, but the fluorescence intensity should be quantified using a fluorimeter or flow cytometer for more accurate analysis. Moreover, since the cells were clearly able to grow (which indicates viability), it is not convincing that C. bungei has a significant impact on rice fermentation. This aspect should be explored further in the discussion section.

8. The sensory data were inadequately discussed in the manuscript. I suggest that the authors provide a more comprehensive analysis and interpretation of the sensory data to enhance the manuscript's overall discussion.

The English language in the manuscript could benefit from improvement. I strongly recommend that the authors seek assistance from a native English speaker to revise the manuscript and ensure its linguistic accuracy. This will help to enhance the overall quality of the manuscript and make it more accessible to a wider audience.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report (Previous Reviewer 2)

The authors have revised the manuscript accordingly. 

Minor editing of English language required

Author Response

Comment 1. Minor editing of English language required.

Answer: Thanks very much! We have carefully checked the manuscript and some errors have been corrected.

Reviewer 3 Report (New Reviewer)

The manuscript was significantly improved. However, some important isseus remain.

1. The use of the Duncan post hoc test did not address my initial question regarding the selection of ANOVA as the appropriate statistical test. Prior to deciding on the analytical methods (parametric or non-parametric) for comparing different groups, it is essential for the authors to first assess the normality of the data. If the data generated from the experiments exhibit a normal distribution, parametric tests like ANOVA can be employed. On the other hand, non-parametric tests are necessary when the data does not follow a normal distribution. This step is crucial in any data analysis, and the authors should clarify whether they conducted a normality test (and specify which one) and describe the methodology used in the Materials and Methods section.

2. The description of the correlation analysis is missing from the Material and Methods section. It is important for the authors to specify which correlation method was utilized, such as Pearson, Spearman, or Kendall. The correlation analysis should be included in section 2.13 of the manuscript. Furthermore, it is necessary for the authors to present the correlation matrix resulting from the analysis.

Author Response

Comment 1: The use of the Duncan post hoc test did not address my initial question regarding the selection of ANOVA as the appropriate statistical test. Prior to deciding on the analytical methods (parametric or non-parametric) for comparing different groups, it is essential for the authors to first assess the normality of the data. If the data generated from the experiments exhibit a normal distribution, parametric tests like ANOVA can be employed. On the other hand, non-parametric tests are necessary when the data does not follow a normal distribution. This step is crucial in any data analysis, and the authors should clarify whether they conducted a normality test (and specify which one) and describe the methodology used in the Materials and Methods section.

Authors’ response: We are very sorry that We did not address your question. The Shapiro-Wilke method was used to test the normal distribution of the data, which has been added in the revised manuscript.

 

Comment 2. The description of the correlation analysis is missing from the Material and Methods section. It is important for the authors to specify which correlation method was utilized, such as Pearson, Spearman, or Kendall. The correlation analysis should be included in section 2.13 of the manuscript. Furthermore, it is necessary for the authors to present the correlation matrix resulting from the analysis.

Authors’ response: Thanks very mush. Pearson method was used for the correlation analysis, and the method and correlation matrix (Table 3) have been added in the revised manuscript.

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

In my opininion the paper is clearly written and reports interesting data on the addition of Cynanchum bungei extracts in yellow wine.

Specific comments:

- line 59: define DNSO

- line 60: “YPD media test” should be “YPD media to test”?

- line 62: “the above groups” is refered to what?

- lines 62 and 91: specify that yellow wine yeast is Saccharomyces cerevisiae

- line 68: define DNS

- lines 80, 82: define PBS and FDA

- line 88: reference [17] is reported after [22], not in numerical order

- line 90: red wine koji is a starter colture?

- line 103: national standard of People’s Republic of China is not reported in REFERENCES section

- lines 115-116: “the analytical sample by filtering” the verb is missing

- lines 133, 135: define DPPH and ABTS

- lines 267-268: it’s not clear to me the meaning of this sentence

- line 348: “…are demonstrated different flavour…” what is the meaning?

- line 355: Saccharomyces

Figure 1B: what is the legend of y-axis?

Figure 4: the vertical bar (on the top right) 0 to 1.5 refers to which unit?

REFERENCES: I suggest to reduce references number

Author Response

In my opininion the paper is clearly written and reports interesting data on the addition of Cynanchum bungei extracts in yellow wine.

Specific comments:

Comment 1: - line 59: define DNSO

The author’ response: I am very sorry the wrong typewriting! It is DMSO and the full name has been presented in the revised manuscript.

 

Comment 2: - line 60: “YPD media test” should be “YPD media to test”?

The author’ response: Thanks very much! The mistake has been corrected as suggested in the revised manuscript.

 

Comment 3: - line 62: “the above groups” is refered to what?

The author’ response: Thanks! “the above groups” is referred to the groups containing Cynanchum bungei Decne extract equivalent to 0%, 5%, 15%, and 25% fresh Cynanchum bungei Decne. It has been revised to “all groups”.

 

Comment 4: - lines 62 and 91: specify that yellow wine yeast is Saccharomyces cerevisiae

The author’ response: Thanks very much! The revision has been performed as suggested.

 

Comment 5: - line 68: define DNS

The author’ response: Thanks! The full name of DNS has been presented in the revised manuscript.

 

Comment 6: - lines 80, 82: define PBS and FDA

The author’ response: Thanks very much! The full name of DNS has been presented in the revised manuscript.

 

Comment 7: - line 88: reference [17] is reported after [22], not in numerical order

The author’ response: Thanks very much! The references and their order have been revised in the revised manuscript.

 

Comment 8: - line 90: red wine koji is a starter culture?

The author’ response: Thanks very much! The starter is a koji (Qu) which is self-produced by Monascus purpureus CGMCC 16790. The related content has been revised.

 

Comment 9: - line 103: national standard of People’s Republic of China is not reported in REFERENCES section

The author’ response: Thanks! The standard has been included in the references.

 

Comment 10: - lines 115-116: “the analytical sample by filtering” the verb is missing

The author’ response: Thanks very much! The sentence has been revised to “the analytical sample was filtered” in the revised manuscript.

 

Comment 11: - lines 133, 135: define DPPH and ABTS

The author’ response: Thanks very much! The full names of DPPH and ABTS have been presented in the revised manuscript.

 

Comment 12: - lines 267-268: it’s not clear to me the meaning of this sentence

The author’ response: Thanks! The related sentence has been re-written in the revised manuscript.

 

Comment 13: - line 348: “…are demonstrated different flavour…” what is the meaning?

The author’ response: Thanks! The sentence has been revised to “ethyl phenylacetate (honey aroma), and octanoic acid ethyl ester (typical fruit and brandy aroma) exhibit different flavour”.

 

Comment 14: - line 355: Saccharomyces

The author’ response: Thanks! “saccharomyces” has been revised to “yeasts”.

 

Comment 15: Figure 1B: what is the legend of y-axis?

The author’ response: Sorry! The y-axis of Figure 1B was covered by Figure 1A. Revision has been performed in the revised manuscript.

 

Comment 16: Figure 4: the vertical bar (on the top right) 0 to 1.5 refers to which unit?

The author’ response: Thanks very much for your careful check! The unit (μg mL^-1) has been added in the figure caption.

 

Comment 17: REFERENCES: I suggest to reduce references number

The author’ response: Thanks very much! The references have been reduced to 44.

Reviewer 2 Report

This manuscript mainly discussed the addition of Cynanchum bungei Decne on the quality of yellow wine, only a small part was related to the vitality of yeast. Normally, authors should track the variation of strain activity in the fermentation process. The activity of yeast includes vitality and viability. Both parameters affect the fermentation ability of the strain and the quality of final product. As shown in figure 1A, the glucose uptake of sample group was different from the control group, which means there should be difference regarding viability and other activities of yeast strain. Although with the same vitality, there would be difference in viability and fermentation performance of yeast during wine fermentation. Hence, current results can not convince me that Cynanchum bungei Decne did not affect the performance of yeast during fermentation. 

Authors better change the title of this manuscript. 

Minors:

1.     Please specify the ripeness，Picking location (longitude and latitude) of Cynanchum bungei Decne

2.     Figure1B, where is the labeling of the Y-axis? 

 

 

 The language should be further improved.

Author Response

Comment 1: This manuscript mainly discussed the addition of Cynanchum bungei Decne on the quality of yellow wine, only a small part was related to the vitality of yeast. Normally, authors should track the variation of strain activity in the fermentation process. The activity of yeast includes vitality and viability. Both parameters affect the fermentation ability of the strain and the quality of final product. As shown in figure 1A, the glucose uptake of sample group was different from the control group, which means there should be difference regarding viability and other activities of yeast strain. Although with the same vitality, there would be difference in viability and fermentation performance of yeast during wine fermentation. Hence, current results can not convince me that Cynanchum bungei Decne did not affect the performance of yeast during fermentation.

The author’ response: Thanks very much! To improve the manuscript, the changes of growth, cell viability, alcohol dehydrogenase activity and glucose uptake ability of yeast were monitored, and the data have been added in the revised manuscript.

 

Comment 2: Authors better change the title of this manuscript.

The author’ response: Thanks very much! The title has been changed to “Effects of Cynanchum bungei Decne addition on the physico-chemical properties and antioxidant activity of rice wine”.

 

Comment 3: Minors:

1. Please specify the ripeness, picking location (longitude and latitude) of Cynanchum bungei Decne
2. Figure1B, where is the labeling of the Y-axis?

The author’ response: Thanks! (1) The ripeness, picking location (longitude and latitude) of Cynanchum bungei Decne have been added as suggested.

(2) The y-axis of Figure 1B was covered by Figure 1A. Revision has been performed in the revised manuscript.