CLA-Producing Probiotics for the Development of a Yogurt-Type Beverage

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. Line 12-13: The line is too long, complex and difficult to understand. Providing you with an example to rewrite the line something like this or break down it into simple, small sentences for better readability and understanding. "This study evaluated the ability of four probiotic strains—Lactobacillus rhamnosus LbRE-LSAS, Bifidobacterium animalis subsp. lactis Bb12, and two yogurt starters (TA040 and LB340)—to ferment MRS medium and milk supplemented with linoleic acid (0, 0.5, or 1 mg/mL). The goal was to produce a yogurt-type beverage enriched with conjugated linoleic acid (CLA) isomers."
2. Line 17: Mentioned " GC chromatography", but the full form of C in GC is chromatography. Please remove the word chromatography and replace it with word analysis. 
3. Line 47-49: The author highlighted the multiple positive health benefits of CLA, but the line seems vague, as the line only talks about the multiple health benefits but doesn't say which health benefits. Please specify the particular health benefits CLA provide for human health. 
4. Line 217: What is the meaning of good biomasses here? Please clarify 
5. Line 223: Needs a clearer explanation of why variation occurs.
6. Figure 1: Statistical significance is not clearly marked in the figure legend. Also, they are here and there. Please do the correct formatting and add the p-value in the figure title. 
7. In both the figures, please correct the decimal number system. Instead of comma, put the dot for representing the values in decimal. 
8. Figure 2: Include p-values or confidence intervals, also correct the formatting of the figure in terms of superscribing the significant values. 
9. Line 294-297: There is no error margin (±SD) reported for CLA conversion rates.
10. Table 3a, 3b: Report p-values or confidence intervals to confirm effects are not random. 

Author Response

Detailed responses to Beverages

 Journal Editor and Referees

 Dear Editor,

First of all, we would like to thank you for the reviewer’s detailed comments and the constructive criticism that has helped to improve the manuscript. A marked copy of the manuscript has been uploaded for the reviewers and editors. We took into account all these comments for the revision of our manuscript and we believe that it significantly strengthened our manuscript. All authors have read and agreed to the revised version of the manuscript.

In the responses to reviewers file, we have included the original comments from the three referees and we developed a point-by-point response in red italics to each of the reviewer’s comment as expected by the journal Editorial committee. Therefore, we believe that we addressed the different issues which were raised by the reviewers.

 In response to comments by Reviewer 1

Comments and Suggestions for Authors

1. Line 12-13: The line is too long, complex and difficult to understand.

 Providing you with an example to rewrite the line something like this or break down it into simple, small sentences for better readability and understanding. "This study evaluated the ability of four probiotic strains—Lactobacillus rhamnosus LbRE-LSAS, Bifidobacterium animalis subsp. lactis Bb12, and two yogurt starters (TA040 and LB340)—to ferment MRS medium and milk supplemented with linoleic acid (0, 0.5, or 1 mg/mL). The goal was to produce a yogurt-type beverage enriched with conjugated linoleic acid (CLA) isomers."

This has been corrected in the revised version.

2. Line 17: Mentioned " GC chromatography", but the full form of C in GC is chromatography. Please remove the word chromatography and replace it with word analysis. 

This has been corrected in the revised version.

3. Line 47-49: The author highlighted the multiple positive health benefits of CLA, but the line seems vague, as the line only talks about the multiple health benefits but doesn't say which health benefits. Please specify the particular health benefits CLA provide for human health. 

This has been corrected in the revised version.

4. Line 217: What is the meaning of good biomasses here? Please clarify 

The adjective “good” was replaced with another one.

5. Line 223: Needs a clearer explanation of why variation occurs.

In general, growth capacity of LAB is not only dependent on medium composition but also strain’s specific (Ziar et al., 2014).

Ziar, H.; Gérard, P.; Riazi, A. Effect of prebiotic carbohydrates on growth, bile survival and cholesterol uptake abilities of dairy-related bacteria. J. Sci. Food Agric. 2014, 94, 1184–1190.

6. Figure 1: Statistical significance is not clearly marked in the figure legend. Also, they are here and there. Please do the correct formatting and add the p-value in the figure title. 

The next paragraph was added to the figure title:

Different lowercase letters (a–c) stand for significantly various mean values of bacterial growth per one strain according to performed one-way ANOVA with the post hoc test (p > 0.05).

7. In both the figures, please correct the decimal number system. Instead of comma, put the dot for representing the values in decimal.

This has been corrected in the revised version.

8. Figure 2: Include p-values or confidence intervals, also correct the formatting of the figure in terms of superscribing the significant values.

This has been corrected in the revised version.

9. Line 294-297: There is no error margin (±SD) reported for CLA conversion rates.

This has been corrected in the revised version.

10. Table 3a, 3b: Report p-values or confidence intervals to confirm effects are not random.

This has been corrected in the revised version.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

After evaluating the article, you should consider the following suggestions.

Abstract

L12 - Write the full name and the abbreviation MRS in brackets in the first place where it is mentioned and then use only the abbreviation.

L17 – Write the full name and the abbreviation GC in brackets in the first place where it is mentioned and then use only the abbreviation.

1. Introduction

L47-L71 - More recent studies, particularly clinical trials, should be included to support the health benefits of CLA.

2. Materials and Methods

L75 – It should be mentioned in brackets where the collection comes from (LMBAFS, Abdelhamid Ibn Badis University, Algeria).

L86 – LA if it has already been abbreviated, then the abbreviation should be used.

L90 – UHT trebuie detaliat

L91 - The sentence should be reworded to avoid the confusion that M17 is MRS.

L100 - "wtw" should be capitalised.

L111 - Why is the purity of the compound written in brackets in some cases and outside the brackets in others? The same spelling should be maintained.

3. Results and Discussion

L169 - The subchapter should be reworded. For example, Bacterial conversion of linoleic acid into CLA.

L184 – L204 The spaces between the figure and its legend should be removed.

L208 - Figures 1a and 1b are mentioned in the legend to figure 1, but do not appear in the figure itself. They only appear in figure 2.

L312 - A comparative graph summarising the CLA conversion rates for each strain would improve clarity.

L332 - The statements could be better supported by citing current clinical studies.

L396 - In some paragraphs, Lactobacilli and Bifidobacteria are not italicised. They should be italicised everywhere.

L410-L413 - No indication whether a positive control (a standard antibiotic) was used for comparison in the antimicrobial tests. Without such a control, the comparison with the antibacterial effect of CLA is incomplete.

L440 – L452 - Units of measurement in tables 3a and 3b should be given at the top of the columns and not in the table title to comply with data reporting standards.

L426 – 427 - Tables 3a and 3b show the association of Streptococcus thermophilus strain TA040 with Lactobacillus delbrueckii ssp. bulgaricus LB340 leads to a decrease in the antibacterial activity of Streptococcus thermophilus TA040. This should be mentioned.

4. Conclusions

L478- L484 - The conclusions should be more detailed and a paragraph with process limitations and future research should be added.

Best regards,

Author Response

Detailed responses to Beverages

 Journal Editor and Referees

Dear Editor,

First of all, we would like to thank you for the reviewer’s detailed comments and the constructive criticism that has helped to improve the manuscript. A marked copy of the manuscript has been uploaded for the reviewers and editors. We took into account all these comments for the revision of our manuscript and we believe that it significantly strengthened our manuscript. All authors have read and agreed to the revised version of the manuscript.

In the responses to reviewers file, we have included the original comments from the three referees and we developed a point-by-point response in red italics to each of the reviewer’s comment as expected by the journal Editorial committee. Therefore, we believe that we addressed the different issues which were raised by the reviewers.

In response to comments by Reviewer  2

Dear Authors,

After evaluating the article, you should consider the following suggestions.

Abstract

L12 - Write the full name and the abbreviation MRS in brackets in the first place where it is mentioned and then use only the abbreviation.

This has been corrected in the revised version.

L17 – Write the full name and the abbreviation GC in brackets in the first place where it is mentioned and then use only the abbreviation.

This has been corrected in the revised version.

1. Introduction

L47-L71 - More recent studies, particularly clinical trials, should be included to support the health benefits of CLA.

Scientific report of EFSA detailed all clinical trials. Health claims were detailed.

1. Materials and Methods

L75 – It should be mentioned in brackets where the collection comes from (LMBAFS, Abdelhamid Ibn Badis University, Algeria).

This has been corrected in the revised version.

L86 – LA if it has already been abbreviated, then the abbreviation should be used.

This has been corrected in the revised version.

L90 – UHT trebuie detaliat

This has been corrected in the revised version.

L91 - The sentence should be reworded to avoid the confusion that M17 is MRS.

This has been corrected in the revised version.

L100 - "wtw" should be capitalised.

This has been corrected in the revised version.

L111 - Why is the purity of the compound written in brackets in some cases and outside the brackets in others? The same spelling should be maintained.

This has been corrected in the revised version.

1. Results and Discussion

L169 - The subchapter should be reworded. For example, Bacterial conversion of linoleic acid into CLA.

This was the authors’ choice for more attractive and illustrative head title.

L184 – L204 The spaces between the figure and its legend should be removed.

This has been corrected in the revised version.

L208 - Figures 1a and 1b are mentioned in the legend to figure 1, but do not appear in the figure itself. They only appear in figure 2.

Both exist. Newer version was added.

L312 - A comparative graph summarising the CLA conversion rates for each strain would improve clarity.

Thank you for the idea. However, all authors judged the data included in the manuscript sufficient.

L332 - The statements could be better supported by citing current clinical studies.

With the due respect to the referee and even if CLA are already in the market, the controversial about their health effect still exists. That’s why including clinical studies for consolidating statement would not be for interest for this in vitro study.

L396 - In some paragraphs, Lactobacilli and Bifidobacteria are not italicised. They should be italicised everywhere.

This has been corrected in the revised version.

L410-L413 - No indication whether a positive control (a standard antibiotic) was used for comparison in the antimicrobial tests. Without such a control, the comparison with the antibacterial effect of CLA is incomplete.

The antibiotic Azithromycin (10µg/disk) was used against Staphylococcus aureus ATCC 29523 = 23 mm, and Escherichia coli ATCC 25922 = 31 mm

Cefoperazone was used against Salmonella typhimirium ATCC 14028: 29 mm

L440 – L452 - Units of measurement in tables 3a and 3b should be given at the top of the columns and not in the table title to comply with data reporting standards.

This has been corrected in the revised version.

L426 – 427 - Tables 3a and 3b show the association of Streptococcus thermophilus strain TA040 with Lactobacillus delbrueckii ssp. bulgaricus LB340 leads to a decrease in the antibacterial activity of Streptococcus thermophilus TA040. This should be mentioned.

Thank you for the remark, we added the paragraph.

1. Conclusions

L478- L484 - The conclusions should be more detailed and a paragraph with process limitations and future research should be added.

This has been corrected in the revised version.

Best regards,

Reviewer 3 Report

Comments and Suggestions for Authors

In abstract or where for the first time appears write full name of MRS.

In the abstract authors need to add which what strains they have the best results.

To my opinion abstract need to be more consist of and give results with numbers of experimentational part.

In preparation of yogurt-type beverage authors need to write full methods of preparation of the sample.

Authors need to cheek statistical results in figure 1 because in my opinion they have really big standard deviation in 0% LA with L.rhamnosus and its very close to the  next B.lactis Bb12 (0% LA).

In description of Figure 1 have a) and b) but in my version I didn’t see figure 1b) please cheek.

The conclusion needs to be write with more results.

Author Response

Detailed responses to Beverages

 Journal Editor and Referees

Dear Editor,

First of all, we would like to thank you for the reviewer’s detailed comments and the constructive criticism that has helped to improve the manuscript. A marked copy of the manuscript has been uploaded for the reviewers and editors. We took into account all these comments for the revision of our manuscript and we believe that it significantly strengthened our manuscript. All authors have read and agreed to the revised version of the manuscript.

In the responses to reviewers file, we have included the original comments from the three referees and we developed a point-by-point response in red italics to each of the reviewer’s comment as expected by the journal Editorial committee. Therefore, we believe that we addressed the different issues which were raised by the reviewers.

In response to comments by Reviewer 3

In abstract or where for the first time appears write full name of MRS.

This has been corrected in the revised version.

In the abstract authors need to add which what strains they have the best results.

All strains were interesting. We respected the 200 words recommended in the journal’s template.

To my opinion abstract need to be more consist of and give results with numbers of experimentational part.

We respected the 200 words recommended in the journal’s template.

In preparation of yogurt-type beverage authors need to write full methods of preparation of the sample.

All the needed detail was given.

Authors need to cheek statistical results in figure 1 because in my opinion they have really big standard deviation in 0% LA with L.rhamnosus and its very close to the  next B.lactis Bb12 (0% LA).

This SD is particularly from different experiments (n=12).

In description of Figure 1 have a) and b) but in my version I didn’t see figure 1b) please cheek.

Both exist. A newer version was added.

The conclusion needs to be write with more results.

This has been corrected in the revised version.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Thank you for revising the manuscript according to the suggested changes.

Author Response

Respected colleague,
We value the work you put into reviewing the manuscript.
Regards

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

The article can be published in its current form as soon as the following minor comments have been clarified.

Abstract

L13 - The first place where MRS was mentioned was in the abstract at L13. At this point, the name MRS should be mentioned in detail and the abbreviation is mentioned in brackets, not at L75.

Results and Discussion

L348 - The statements will be more convincingly supported if the human health benefits are presented in accordance with the cited references.

Conclusions

L500 - The conclusions should include future research.

Best regards,

Author Response

Respected colleague,
We value the work you put into reviewing the manuscript.
Regards

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

I accept the corrections and have no additional comments to the authors.

Author Response

Respected colleague,
We value the work you put into reviewing the manuscript.
Regards