Enhancing Steamed Bread Quality Through Co-Fermentation of Sourdough with Kazachstania humilis and Lactobacillus plantarum
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis study aims to evaluate the effects of co-fermentation in bread making. While the work doesn't introduce entirely new concepts, it is presented clearly. Including more comparisons with existing literature and conducting a statistical analysis could significantly enhance the quality of the study.
Below are some additional considerations
Abstract
The abstract provides comprehensive information about the work conducted.
Introduction
The introduction is well-written and effectively presents the background of the study.
Materials and Methods
Statistical analysis is missing
While I appreciate the flowchart in section 2.2, it would be more appropriate to introduce it with a brief explanation rather than placing it in a separate paragraph. The authors could consider including an introductory paragraph for section 2.2, where the flowchart is presented, and then adding sub-sections (such as 2.2.1, 2.2.2, etc.) for detailed descriptions of the methods used.
Line 107: Do the authors mean 12 hours of pre-culture in MRS and YPD? If so, why do they talk about 24h in paragraph 2.3?
Section 2.4 a diagram might help to understand
Line 206: do the authors mean “20 hours” (instead of days)
Results
A more thorough comparison with existing literature could enhance the work.
Figure 2(a). I can't find a significant difference between the two trends. Did the authors perform a statistical evaluation to highlight possible differences?
Figure 5 the legend does not seem exhaustive and should include more information
Author Response
Comment 1: Statistical analysis is missing
Response: Thank you for pointing this out. We agree with this comment. Therefore, we have added this part on page 4 paragraph 3 line 140-146
Comment 2: While I appreciate the flowchart in section 2.2, it would be more appropriate to introduce it with a brief explanation rather than placing it in a separate paragraph. The authors could consider including an introductory paragraph for section 2.2, where the flowchart is presented, and then adding sub-sections (such as 2.2.1, 2.2.2, etc.) for detailed descriptions of the methods used.:
Response: Thank you for this insight we agree to the comment and have added the introductory paragraph on section 2.2 on page 4 and added the subsections as advised on page 4 paragraph 4 line 148-151 and the sub sections 2,2.1 and 2.2.2 on pages 5 and 6 lines 154 and 175 respectively.
Comment 3: Line 107: Do the authors mean 12 hours of pre-culture in MRS and YPD? If so, why do they talk about 24h in paragraph 2.3?
Response: We appreciate you noticing that error. we have corrected the error and changed the time to 24h which is the correct time for pre-culture .we have made changes on page 5 paragraph 2 line 156.
comment 4: Section 2.4 a diagram might help to understand.
Response: Thank you for pointing this out we agree to the insight and have added a diagram on section 2.4 in page 6 paragraph 1 line 173-174
comment 5:Line 206: do the authors mean “20 hours” (instead of days)
Response: Thank you for noticing this. we agree to the comment and have corrected the mistake on page 9 paragraph 1 line 277
Comment 6 :Results. A more thorough comparison with existing literature could enhance the work
Response : Thank you for this comment we agree to this insight and have added literature section on page 8 paragraph 1 line 242-253
Comment 7: Figure 2(a). I can't find a significant difference between the two trends. Did the authors perform a statistical evaluation to highlight possible differences?
Response: Thank you for your insightful comment. We acknowledge that the differences in colony counts between LP and LP+KH groups in Figure 4 (a) appear subtle. We performed statistical analysis using one-way ANOVA at each time point to assess the significance of the differences between the two groups. Although the trends suggest enhanced LP growth in the LP+KH group during the late fermentation phase, the differences were not statistically significant (p > 0.05), except at the 20-hour point, where a significant difference was observed (p < 0.05). We have now added this clarification in the revised figure legend and Results section on page 10 line 298-301
comment 8: Figure 5 the legend does not seem exhaustive and should include more information.
Response: We agree to this. We have updated the legend to be more exhaustive with more information in page 13 paragraph 1 line 376 and updated to figure 7.
Reviewer 2 Report
Comments and Suggestions for AuthorsPeer-Review Comments
General Comments:
The manuscript titled "Enhancing Steamed Bread Quality through Co-Fermentation of Sourdough with Kazachstania humilis and Lactobacillus plantarum" presents an interesting and well-conducted study. The experimental design is coherent, and the conclusions are consistent with the data presented. The co-fermentation strategy with Kazachstania humilis and Lactobacillus plantarum appears promising for improving the sensory and textural quality of steamed bread. The manuscript is overall well-structured and written clearly. However, I have some important suggestions and specific comments that need to be addressed before acceptance:
Culture Media Description:
The manuscript mentions the use of MRS and YPD media but does not provide full compositions or detailed descriptions.
Please include the complete formulation or the main composition of both MRS and YPD media, or at least a reference describing them, to ensure reproducibility.
Selective Cultivation and Antibiotics:
It is important to highlight that both MRS and YPD media can support the growth of both bacteria and yeasts if selective agents are not used.
In order to accurately quantify Lactobacillus plantarum and Kazachstania humilis separately during plating, selective antibiotics (such as cycloheximide for inhibiting yeast in MRS, and chloramphenicol for inhibiting bacteria in YPD) should be added to the media.
Please clarify whether antibiotics were used in your plating methods. If not, this is a significant methodological limitation that should be discussed.
If antibiotics were not added, it must be acknowledged that the plate counts may not fully distinguish between bacteria and yeasts, potentially affecting the results' accuracy.
The Sensory Analysis section requires the inclusion of the Ethics Committee approval number that authorized the study. It is not permissible to publish sensory analysis results without prior approval from an Ethics Committee. A description of the Ethics Committee’s approval must be incorporated into the methodology. Additionally, clarify whether the panelists signed a "Informed Consent Form" regarding their participation. Was the data collected anonymously? If anonymity was not guaranteed, explicit approval from the panelists for publication is necessary. Please describe in the methodology how these ethical procedures were conducted, in accordance with the guidelines for research involving human subjects.
Figures 1 and 2 (growth curves and viable counts) should clearly state whether selective media were used for separate enumeration of bacteria and yeast. This is crucial for validating the claims made regarding microbial growth dynamics.
I suggest adding a section discussing the limitations of the study, the potential for industrial application, and future perspectives for scaling up the research to pilot and industrial levels.
Comments on the Quality of English Language
Minor English editing is suggested throughout the manuscript to improve flow (e.g., "was" should be corrected to "were" in "LAB species (Lactobacillus plantarum1 (LP1)) and the yeast species (Kazachstania humilis strain (KH2-1)) was used").
There are several typos where words are merged (e.g., “KH sourdough was prepared by mixing 100 g of wheat flour(strong flour, Wudeli Flour Group…” → there should be a space after "flour").
Author Response
Comment 1:The manuscript mentions the use of MRS and YPD media but does not provide full compositions or detailed descriptions. Please include the complete formulation or the main composition of both MRS and YPD media, or at least a reference describing them, to ensure reproducibility.
Response: Thank you for pointing this out. We agree with this comment. Therefore, we have added this part on page 3 and 4 paragraph 2 lines 114-139
Comment 2: Selective Cultivation and Antibiotics: It is important to highlight that both MRS and YPD media can support the growth of both bacteria and yeasts if selective agents are not used. Page 3 paragraph 2 line 110-113
Response: Thank you for this insight we agree to the comment and have highlighted that selective agents were used on pages 3 line110-113.
Comment 3: In order to accurately quantify Lactobacillus plantarum and Kazachstania humilis separately during plating, selective antibiotics (such as c-0ycloheximide for inhibiting yeast in MRS, and chloramphenicol for inhibiting bacteria in YPD) should be added to the media.
Response: : Thank you for pointing this out we agree to the insight and highlighted that c-0ycloheximide and chloramphenicol were added to the media during plating process as highlighted in Page 3 paragraph 2 line 110-113
comment 4: Please clarify whether antibiotics were used in your plating methods. If not, this is a significant methodological limitation that should be discussed.
If antibiotics were not added, it must be acknowledged that the plate counts may not fully distinguish between bacteria and yeasts, potentially affecting the results' accuracy.
Response Thank you for this insight we agree to the comment and have highlighted that antibiotics were added to the media as shown in Page 3 paragraph 4 line 122 and page 4 paragraph 2 line 134
comment 5: The Sensory Analysis section requires the inclusion of the Ethics Committee approval number that authorized the study. It is not permissible to publish sensory analysis results without prior approval from an Ethics Committee. A description of the Ethics Committee’s approval must be incorporated into the methodology. Additionally, clarify whether the panelists signed a "Informed Consent Form" regarding their participation. Was the data collected anonymously? If anonymity was not guaranteed, explicit approval from the panelists for publication is necessary. Please describe in the methodology how these ethical procedures were conducted, in accordance with the guidelines for research involving human subjects.
Response: Thank you for highlighting this. We agree that sensory Analysis section requires the inclusion and we have included it on page 3 paragraph 1 line 93-100
Comment 6 : Figures 1 and 2 (growth curves and viable counts) should clearly state whether selective media were used for separate enumeration of bacteria and yeast. This is crucial for validating the claims made regarding microbial growth dynamics.
Response : Thank you for this comment we agree to this insight and have highlighted this on page 8 paragraph 2 line 261 and page 9 paragraph 1 line 284.
Comment 7: I suggest adding a section discussing the limitations of the study, the potential for industrial application, and future perspectives for scaling up the research to pilot and industrial levels.
Response: Thank you for your insightful comment and we have added that section on page 15 paragraph 2 line 450-461
comment 8: Minor English editing is suggested throughout the manuscript to improve flow (e.g., "was" should be corrected to "were" in "LAB species (Lactobacillus plantarum1 (LP1)) and the yeast species (Kazachstania humilis strain (KH2-1)) was used").
There are several typos where words are merged (e.g., “KH sourdough was prepared by mixing 100 g of wheat flour (strong flour, Wudeli Flour Group…” → there should be a space after "flour").
Response: We agree to this. We have corrected the minor English editing errors in as shown in on page 3 paragraph 3 line 103 and on page 5 paragraph 2 line 164 as well as the entire manuscript.
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors have addressed all the suggested revisions. In my opinion, the manuscript is now suitable for publication in Fermentation.
