Correlation between the Characteristic Flavour and Microbial Community of Xuanwei Ham after Ripening

Response to Reviewer 1 Comments

（Changes in the article are marked in yellow for quick review by reviewers）

1. Summary

Thank you for your letter and the reviewers’ comments concerning our manuscript Fermentation-3034725 entitled “Correlation between the characteristic flavour and microbial community of Xuanwei ham after ripening”.

All the comments were valuable and very helpful for revising and improving our paper and the important guide to our research. We have studied the comments carefully and have corrected them in our paper accordingly. We hope that the revised manuscript will meet the standards for publication in your journal.

We are looking forward to hearing from you soon.

Sincerely yours,

Guipeng Li, Master

Faculty of Food Science

Tibet Agriculture & Animal Husbandry University, Linzhi 860000, P.R. China

Tel.: 17393197853

E-mail: [email protected]

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Can be improved

Agree

Is the research design appropriate?

Can be improved

Agree

Are the methods adequately described?

Can be improved

Agree

Are the results clearly presented?

Can be improved

Agree

Are the conclusions supported by the results?

Can be improved

Agree

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: Abstract (Lines 15-31): The abstract is concise but could include brief information about the major findings to give readers a quick overview of the results.

Response 1: Thank you for your valuable advice. We agree with this comment. To make the reader quickly understand the results, we have rewritten the important parts of the summary carefully and removed the unimportant information. (Detailed changes are found on lines 15-30.)

Comments 2: Introduction (Lines 34-75): Add more recent references and compare the study's approach and findings with other studies in the field. The introduction could benefit from a more comprehensive literature review. While it cites relevant studies, it would be beneficial to include more recent research and compare findings with similar studies on other types of fermented meat products.

Response 2: Thank you for your guidance, and we think your suggestion is very necessary, So we added some recent references and compared them with other research in the field, the new additions are presented in lines 42-46 and 61-66, and the related references are presented separately in lines 431-432 and 458-459.

Comments 3: Materials and Methods (Lines 76-175): Provide a more detailed explanation of the sampling method and the statistical tools used for data analysis. Sampling Procedure: The procedure for sampling could be described in more detail. Specifically, the selection process for the three hams each year and any potential biases should be addressed. Statistical Analysis: While heatmap analysis is mentioned, a detailed description of the statistical methods used to analyze the data (e.g., ANOVA, correlation coefficients) is missing.

Response 3: We appreciate your valuable advice very much.

1. The ham selection process included the following

 ①Background Research: Before selecting the hams, a thorough background research was conducted to understand the different types of Xuanwei hams and their characteristics and production process. The final selection was made to Yunnan Yiji Food Co. ②Sample Selection: Ensure that each sample of Xuanwei ham selected was representative and from different batches or sources to reduce possible chance and bias. ③Randomisation: Randomisation of ham samples was carried out to ensure the reliability and statistical significance of the experimental results. (Lines 83-84)

Potential biases are as follows

①Subjective bias: When evaluating the experimental results, avoid being influenced by subjective bias and try to analyse the data and conclusions as objectively as possible.②Sample bias: Ensure that the selected sample set of hams adequately represents the overall characteristics of Xuanwei hams, and avoid bias in sample selection.③Experimental design: Adopt the method of double-blind experimental design to minimize the possibility of experimental results being influenced by bias.

2. correlation heatmap was completed using the Wekemo Bioincloud (https:// www.bioincloud.tech). Referring to the method of Gao et al [1] with slight modification, the upper limit of the number of relevant features in the abundance table of the genus was set to the first 20 on the platform provided by this company, and the first 20 microorganisms screened for significance (P < 0.05) features were correlated with the environmental factors to do the correlation analysis with the correlation coefficient threshold set to 0.1, and the correlation coefficients with an absolute value greater than the threshold would be marked as significant. （Located in lines 184-190）

[1]       Yunyun Gao, Guoxing Zhang, Shunyao Jiang, Yong-Xin Liu. 2024. Wekemo Bioincloud: A user-friendly platform for meta-omics data analyses. iMeta 3: e175. https://doi.org/10.1002/imt2.175

Comments 4: Results (Lines 176-315): Include more visual data representations and a detailed breakdown of the findings for each year. Detailed Analysis: The discussion on the microbial communities could be more detailed, particularly in explaining why certain genera dominate at specific ripening times.

Response 4: Thank you for your guidance. There are many factors affecting the microorganism of Xuanwei ham, such as environment, temperature, moisture, salt content, etc. The present experiment found that Sarocladium was the dominant genus in W1 and W4. In this experiment, Sarocladium was found to be the dominant genus in W1, W2 and W4, while Vibrio was found to be the dominant genus in W3, which may be due to the fact that as the hams matured, the salt content of the hams gradually decreased, which resulted in a decrease in the inhibitory ability of the microorganisms and thus Vibrio became the dominant genus in W3. However, Sarocladium was the dominant genus in W4, probably because a series of biochemical reactions such as protein hydrolysis, amino acid degradation, lipolysis and lipid oxidation were beneficial to the reproduction of this microorganism, and thus Sarocladium became the dominant genus in W4. In general, the dominant genera of Xuanwei ham in different years were Sarocladium and Vibrio. (Lines 325-335)

Comments 5: Figure 3: In the paragraph between rows 231-239, reference is made to "Figure 3c", "Figure 3d", and "Figure 3e" to describe various VOCs found in Xuanwei hams of different ages. However, there are no figures with these subsections in the PDF document. This could be a numbering error or a missing reference to specific subparts in Figure 3. Check the other figure in the text

Response 5: We are very sorry to make this low-level mistake. We have marked the changed section icon in the text (at lines 221-235)

Comments 6: Discussion and Conclusion: The practical implications of the findings for the production and storage of Xuanwei ham could be elaborated. For instance, how can producers use this information to improve the flavour and quality of the ham? Discussion (Lines 316-415): Expand on the practical applications of the study's findings and discuss potential reasons for the observed microbial community changes. Conclusion (Lines 416-430): Summarize the key findings and their implications more succinctly. Include suggestions for future research directions.

Response 6: Thank you for your guidance, and we think your suggestion is very necessary, I have restated the summary briefly and written the discussion in more detail. Discussion ideas are roughly as follows;

 It is well known that the older the ham, the richer and more flavourful it is, the more popular it is with consumers, but it is also expensive and famous hams are in short supply. （For example, Iberian ham.） This paper aims to study the correlation between individual dominant microorganisms and flavour to screen out microorganisms that are beneficial to flavour, so that they can be inoculated onto unfermented hams to achieve the flavour of aged hams in a short period of time, thus reducing the cost to the producers and making hams more acceptable to a wider range of people. This will require more effort and dedication in the future, but we are confident. (A few sentences of Lines 389-499 are briefly mentioned at the discussion of the article.)

Comments 7: The article contains some grammatical errors and awkward phrasing that could be improved for better readability.

Response 7: We are very sorry that many English grammar in the article is inappropriate due to our language barrier. The English grammar problems and the use of words in the article have been revised repeatedly. If there are still any problems that need to be modified, please feedback to us, and we will make timely modifications.

4. Additional clarifications

We have tried our best to repeatedly modify the basic problems in the article such as formatting problems, picture typesetting, etc. If any problems need to be modified, please feedback to us, and we will revise it in time.

Response to Reviewer 2 Comments

（Changes in the article are marked in light blue for quick review by reviewers）

1. Summary

Thank you for your letter and the reviewers’ comments concerning our manuscript Fermentation-3034725 entitled “Correlation between the characteristic flavour and microbial community of Xuanwei ham after ripening”.

All the comments were valuable and very helpful for revising and improving our paper and the important guide to our research. We have studied the comments carefully and have corrected them in our paper accordingly. We hope that the revised manuscript will meet the standards for publication in your journal.

We are looking forward to hearing from you soon.

Sincerely yours,

Guipeng Li, Master

Faculty of Food Science

Tibet Agriculture & Animal Husbandry University, Linzhi 860000, P.R. China

Tel.: 17393197853

E-mail: [email protected]

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Yes

Is the research design appropriate?

Yes

Are the methods adequately described?

Yes

Are the results clearly presented?

Yes

Are the conclusions supported by the results?

Can be improved

Agree. Detailed changes are listed below

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: Keywords: please indicate keywords that are not present in the title.

Response 1: Thank you for pointing this out. We agree with this comment. We have therefore rewritten individual keywords carefully, changing microorganisms to microbial diversity, flavour to volatile flavour, and filling in keyword free amino acids. (located in line 33 of the article).

Comments 2: Conclusions: I believe that follow-up studies should try to analyse the correlation between major microorganisms and the acceptability of the hams. Please consider this to be included in the conclusions.

Response 2: Thank you for your guidance, and we think your suggestion is very necessary, and it's really inspiring to our team, and it's something we need to work on, We not only need to screen beneficial microorganisms to shorten the fermentation cycle of Xuanwei ham, but also need to use scientific experimental results to make ham producers accept its feasibility. This sentence has been added to the conclusion. (in lines 394-401).

4. Response to Comments on the Quality of English Language

Point 1: There are some typing errors, such as: dif-ferent; relation-ships/chang-es. Please verify throughout the manuscript.

Response 1: We are very sorry that many English grammar in the article is inappropriate due to our language barrier. The English grammar problems and the use of words in the article have been revised repeatedly. If there are still any problems that need to be modified, please feedback to us, and we will make timely modifications.