Identification of Lactic Acid Bacteria on Raw Material for Cocoa Bean Fermentation in the Brazilian Amazon
, Nelson Rosa Ferreira 1,2,*, Maria Beatriz A. Gloria 3, Rubens Menezes Gobira 4, Felipe de Andrade Maia 2 and Alessandra Santos Lopes 1,2,*Round 1
Reviewer 1 Report
The topic is interesting but the paper needs major revision, especially for the results section. The authors should include results about their experiments (PCR verification gels, etc.) to support the results and discussion section.
Besides, is it possible to analyze the molecules generated by fermentation using the identified microorganisms? This will provide further discussion of the results.
Author Response
Dear Editor,
Attached, please find the revised manuscript with requested clarifications and changes. We thank the Editor and reviewers for taking the time and for the careful reading and evaluation this manuscript. We appreciated the suggestions and we are certain that the manuscript has been enhanced with the changes.
Our replies to the recommendations are listed below and indicated in the manuscript with red color.
Best regards.
Reviewer #1
“The topic is interesting but the paper needs major revision, especially for the results section.”
Our reply: The manuscript was revised and improved. Thank you.
“The authors should include results about their experiments (PCR verification gels, etc.) to support the results and discussion section.”
Our reply: This is a good point. We verify the integrity of the PCR products using 1% agarose gel with SYBR® Safe DNA Gel Stain (Invitrogen) after 30 min. of running in 0.5x TBE buffer under 80V electrical conduction.
We did not identify any new species of bacteria or other microorganisms in this research, which would lead to a detailed study of the characteristics obtained in the agarose gel in order to characterize the DNA and/or the PCR product obtained in relation to the applied conditions of the analysis and/or the number of base pairs (bp). The high identification similarity values that we obtained with the sequencing and comparison with genetic databases, reinforce the efficiency of the PCR conditions applied, PCR product purification process and the accuracy of the genetic sequences deposited in Genbank. This led us to dispense the inclusion of the PCR products gel as well as its analysis, which does not compromise the quality of our findings.
“Besides, is it possible to analyze the molecules generated by fermentation using the identified microorganisms? This will provide further discussion of the results.”
Our reply: Our main objective was to identify the lactic acid bacteria (LAB) present at the beginning of the fermentation process in a farm located in a city of great agrarian importance for the Amazon region, Tomé-Açu. We think it is very important to know the metabolic dynamics that the LAB species identified in our study play during the cocoa fermentation process. However, these studies are being developed in other researches in our laboratory, not only with cocoa, but in other fermented foods that are typical of the Brazilian Amazon region too. Thank you for your suggestion.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
These researchers have isolated and identified in the raw material utilized for the cocoa fermentation in a city in the Brazilian Amazon several LAB. Their findings can be an option for future studies on the use of this bacteria group in the starter culture production. I have enjoyed the reading of this communication.
Author Response
Dear Editor,
Attached, please find the revised manuscript with requested clarifications and changes. We thank the Editor and reviewers for taking the time and for the careful reading and evaluation this manuscript. We appreciated the suggestions and we are certain that the manuscript has been enhanced with the changes.
Our replies to the recommendations are listed below and indicated in the manuscript with red color.
Best regards.
Reviewer #2
“These researchers have isolated and identified in the raw material utilized for the cocoa fermentation in a city in the Brazilian Amazon several LAB. Their findings can be an option for future studies on the use of this bacteria group in the starter culture production. I have enjoyed the reading of this communication.”
Our reply: Thank you very much for your positive feedback.
Author Response File: Author Response.pdf
Reviewer 3 Report
This study describes the identification of lactic acid bacteria in raw materials used for cocoa fermentation, thus contributing to the understanding of microbial dynamics and evolution during cocoa fermentation as well as the understanding of microbial dynamics and succession during cocoa fermentation.
1.The introductory section of this paper does not give a completely clear account of the background to the study, this needs to be improved and the author is invited to add a full account.
2.In the section on results and discussion, I think the author should have discussed the results in comparison with previous studies rather than giving a flat narrative.
Author Response
Dear Editor,
Attached, please find the revised manuscript with requested clarifications and changes. We thank the Editor and reviewers for taking the time and for the careful reading and evaluation this manuscript. We appreciated the suggestions and we are certain that the manuscript has been enhanced with the changes.
Our replies to the recommendations are listed below and indicated in the manuscript with red color.
Best regards.
Reviewer #3
“This study describes the identification of lactic acid bacteria in raw materials used for cocoa fermentation, thus contributing to the understanding of microbial dynamics and evolution during cocoa fermentation as well as the understanding of microbial dynamics and succession during cocoa fermentation.”
Our reply: Thank you for your time to evaluate this manuscript.
“1. The introductory section of this paper does not give a completely clear account of the background to the study, this needs to be improved and the author is invited to add a full account.”
Our reply: The introduction section was improved (Pages 1-2, Lines 31-58). Thank you.
“2. In the section on results and discussion, I think the author should have discussed the results in comparison with previous studies rather than giving a flat narrative.”
Our reply: We appreciate your suggestion. The discussion followed a careful search in literature about the lactic acid bacteria found in this study. This section was improved and can be found on Pages 3-5.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The authors addressed well my comments. The communication could be accepted in its current form.
