Screening, Characterization and Probiotic Properties of Selenium-Enriched Lactic Acid Bacteria

Round 1

Reviewer 1 Report

Authors: Lixia Zan, Zhe Chen, Ben Zhang, Xiangyu Zou, Afeng Lan, Wenyi Zhang, Yahong Yuan, Tianli Yue.

Ref: Submission ID Fermentation 2684996

Title:"Screening, characterization and prebiotic properties of selenium-rich lactic acid bacteria”

The aim of this study was to investigate the Se-enriched capabilities of six strains of lactic acid bacteria, aiming to explore the relationship between changes in the bacterial cell structure and Se environmental metabolism in Se-enriched lactic acid bacteria.

This study is interesting, well done with a potential application. However,  I would like to call the authors attention at some points throughout the text. 

1.               In lines 175-179, statistical analysis was mentioned, but I did not see any real data treatment with statistics throughout the manuscript, you have to make this clear.

2.                Line 196, best in impersonal

3.               In my opinion the text in lines 203-206 is not that clear, you should start as a new paragraph in line 204 (Figure 1H…).

4.               I should think that lines 229-234 need a wider description

5.               In line 290, which is the value of the Se concentration threshold?

6.               In my viewpoint, the conclusion section does not seem to be a real conclusion but an extent of the discussion, you have to show what is outstanding from this study. 

Overall the english is fine, with bits and pieces to be checked in the text (punctuation marks).

Author Response

Dear Reviewer:

Thank you for the comments concerning our manuscript entitled "Screening, characterization and probiotic properties of selenium- enriched lactic acid bacteria".

Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:

1. In lines 175-179, statistical analysis was mentioned, but I did not see any real data treatment with statistics throughout the manuscript, you have to make this clear.

Response to comment 1: Thank you for bringing this to our attention. We have supplemented the information on the statistical methods used for data analysis. Additionally, we have modified the presentation of the data in the results section to reflect the format of Mean+SD.

2. Line 196, best in impersonal.

Response to comment 2: Thank you for the valuable suggestion and for helping us improve the quality of our manuscript. In response to this comment, we have revised line 196 to use impersonal language.

3. In my opinion the text in lines 203-206 is not that clear, you should start as a new paragraph in line 204 (Figure 1H…).

Response to comment 3: We thank the reviewer for bringing this to our attention and for helping us improve the organization and clarity of our manuscript. We have made the suggested revision and started a new paragraph at line 204.

4. I should think that lines 229-234 need a wider description.

Response to comment 4: Thank you for your comment regarding the need for a wider description in lines 229-234. We have expanded the description in the 3.3 section to provide a more comprehensive explanation.

5. In line 290, which is the value of the Se concentration threshold?

Response to comment 5: We apologize for the oversight in not providing this information. We have made comprehensive revisions to the discussion section.

6. In my viewpoint, the conclusion section does not seem to be a real conclusion but an extent of the discussion, you have to show what is outstanding from this study. 

Response to comment 6: We appreciate the viewpoint regarding the conclusion section of our manuscript. In response, we have revised the conclusion section to provide a concise summary of the key contributions and outstanding aspects of the study.

Once again, we appreciate your review and guidance, and we will continue to strive for your approval.

Best regards,

Tianli Yue

Reviewer 2 Report

Document is well written, is interesting. Some recommendations could be considered.

Line 44. Se-deficient, not se-deficient.

Line 70. space LABas. "LAB as"

Line 73 and 74. What strain is LA 21805? Could you give gender and species? Italicize scientific names.

Line 91 and 166. What solution is PBS?

Line 95 and 96. Be uniform, use names or chemical formulas. Ej. Argon, hydrochloric acid, and NaBH4 solution

Document is well written

Author Response

Dear Reviewer:

Thank you for the comments concerning our manuscript entitled "Screening, characterization and probiotic properties of selenium- enriched lactic acid bacteria".

Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:

1. Line 44. Se-deficient, not se-deficient.

Response to comment 1: Thank you for pointing that out. We have made the necessary correction in the revised manuscript.

2. Line 70. space LABas. "LAB as"

Response to comment 2: Thank you for pointing out the space issue. We have corrected it to "LAB as."

3. Line 73 and 74. What strain is LA 21805? Could you give gender and species? Italicize scientific names.

Response to comment 3：Thank you for your significant reminding. We have provided the gender and species for LA 21805 and italicized the scientific names.

4. Line 91 and 166. What solution is PBS?

Response to comment 4: We apologize for the oversight, and have clarified that PBS stands for "Phosphate-Buffered Saline."

5. Line 95 and 96. Be uniform, use names or chemical formulas. Ej. Argon, hydrochloric acid, and NaBH4 solution

Response to comment 5: We appreciate your suggestion and have made the appropriate changes to ensure uniformity in the use of names or chemical formulas in the manuscript.

Once again, we appreciate your review and guidance, and we will continue to strive for your approval.

Best regards,

Tianli Yue

Reviewer 3 Report

Title: " Screening, characterization and prebiotic properties of selenium-rich lactic acid bacteria

Manuscript Number: fermentation-2684996

Journal: fermentation

Article Type: Original Article

Comments to the author (if any):

Abstract

The article entitled " Screening, characterization and prebiotic properties of selenium-rich lactic acid bacteria" provided the information of high-selenium strains which were characterized for their structure and probiotic properties. The abstract still need to re-write and improve it. It displayed untidiness. The abstract needs a clearer structure. It would be helpful to present the main points in a more organized manner. For instance, the introduction of the topic, M&M, key findings or contributions, and the focus of the article could be structured more coherently. The isolates were isolated from X, characterized for PGP traits—screened for Y…. Subjected to concentrations... A,, then followed by Results and conclusion. Avoid the repetition.

·       What was the objective the experiment?

·       English. Moreover, the English language used in the article has significant issues and requires extensive editing. The authors must be seeking help from native speaker to improve its quality.

·       For instance, and activated the 3rd generation, precipitated bacterial body or precipitated bacterial mass?

·       Why the authors want to know the effect of Se on bacterial cell? (morphology Structural Characterization of SL)

Introduction

The introduction needs to extend it. The following point must be included.

The role of bacteria in probiotic, industrialization, main source of lactobacillus bacteria, natural source that could be used to get them.

M&M

·       The following section needs to re-write

·       Strains and Reagents

·       Six LAB (LA 21805, Lactobacillus paracasei 20241, Lactobacillus fermentum 21828, Lactobacillus casei 23185, Lactobacillus acidophilus 6064, LA 6076, were purchased, but then what is the point to do Strain Identification by 16S rRNA Sequencing?

Minor

·       Selenium-enriched strains. Used it in the beginning of abstract. How do you know?

·       Write complete name of abbreviation for the first time. MRS, PBS, HS

·       What does it mean, does the strain has copy? between the original strains and selenium-enriched LAB were observed

·       16S rRNA repeated in abstract

·       bacterial DNA extraction kit. Name, company, model

·       Scanning Electron Microscopy (SEM) Analysis and Fourier Transform Infrared (FT-IR) Analysis X-ray Diffraction (XRD) Analysis. Add the references

·       How determined the number of viable bacteria

·       Which test , Tukey or Duncan?

Results

·       They were purchased not n selected?  In this study, six strains of bacteria were selected

·       Figure 1. Which alphabet represent which bacteria? Explain it clearly.  

·       Reconstruct the Phylogenetic tree and differentiate your strains (bold or colr).

·       Why the authors are in rush. They wrote a short caption for Figure 3. They must explain all the important information regarding the figure.

Discussion

·       I could not find any discussion because the authors simply collated information from the literature.

·       The order of the results must have been followed in the discussion.

Conclusion

·       All the findings were repeated in the conclusion. For instance, discussing 16Sr RNA is not necessary.

·       Throughout the research trial, six strains were examined; however, the author concluded that "strain LA 6076 was" chosen.

·       Rewrite it entirely, mentioning just the standout outcomes.

Author Response

Dear Reviewer:

Thank you for the comments concerning our manuscript entitled "Screening, characterization and probiotic properties of selenium- enriched lactic acid bacteria".

Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:

1. Abstract

The article entitled " Screening, characterization and prebiotic properties of selenium-rich lactic acid bacteria" provided the information of high-selenium strains which were characterized for their structure and probiotic properties. The abstract still need to re-write and improve it. It displayed untidiness. The abstract needs a clearer structure. It would be helpful to present the main points in a more organized manner. For instance, the introduction of the topic, M&M, key findings or contributions, and the focus of the article could be structured more coherently. The isolates were isolated from X, characterized for PGP traits—screened for Y…. Subjected to concentrations... A,, then followed by Results and conclusion. Avoid the repetition.

What was the objective the experiment?

English. Moreover, the English language used in the article has significant issues and requires extensive editing. The authors must be seeking help from native speaker to improve its quality.

For instance, and activated the 3rd generation, precipitated bacterial body or precipitated bacterial mass?

Why the authors want to know the effect of Se on bacterial cell? (morphology Structural Characterization of SL)

Response to comment 1:

Thank you for your valuable feedback on our manuscript. We appreciate your suggestions and have made the following revisions in response to your comments:

We have rewritten the abstract to present the main points in a more organized manner, including the introduction of the topic, methodology, key findings, and the focus of the article.

The study investigated the effect of selenium concentration on the growth and enrichment of, and evaluate the probiotic properties of selenium-enriched LAB. The objective of our experiment was to provide scientific evidence for the development of selenium-enriched lactic acid bacteria products by gaining a deeper understanding of the mechanisms of LAB selenium enrichment and adaptation.

We have sought the assistance of a native speaker to extensively edit and improve the quality of the language used in the manuscript.

We have made the necessary corrections, for example, using "bacterial biomass" instead of "precipitated bacterial body" for clarity.

We wanted to know the effect of selenium on bacterial cells because high selenium environments can cause changes in the cell structure and chemical composition of microorganisms. This is important for understanding the adaptability of Lactobacillus plantarum in selenium-rich environments and its potential applications.

2. Introduction

The introduction needs to extend it. The following point must be included. The role of bacteria in probiotic, industrialization, main source of lactobacillus bacteria, natural source that could be used to get them.

Response to comment 2:

Thank you for your valuable feedback on our manuscript. We have carefully considered your comments and made the following revisions:

We have extended the introduction to include the role of bacteria in probiotics, their industrialization, the main sources of lactobacillus bacteria, and natural sources that could be used to obtain them. These additions provide a more comprehensive overview of the context and significance of our study.

3. M&M

The following section needs to re-write

Strains and Reagents

Six LAB (LA 21805, Lactobacillus paracasei 20241, Lactobacillus fermentum 21828, Lactobacillus casei 23185, Lactobacillus acidophilus 6064, LA 6076, were purchased, but then what is the point to do Strain Identification by 16S rRNA Sequencing?

Selenium-enriched strains. Used it in the beginning of abstract. How do you know?

Write complete name of abbreviation for the first time. MRS, PBS, HS

What does it mean, does the strain has copy? between the original strains and selenium-enriched LAB were observed

16S rRNA repeated in abstract

bacterial DNA extraction kit. Name, company, model

Scanning Electron Microscopy (SEM) Analysis and Fourier Transform Infrared (FT-IR) Analysis X-ray Diffraction (XRD) Analysis. Add the references

How determined the number of viable bacteria

Which test , Tukey or Duncan?

Response to comment 3:

Thank you very much for your valuable feedback on our manuscript. We have made modifications and provided responses to your suggestions:

We have clarified the purpose of strain identification by 16S rRNA sequencing in the manuscript. This step was conducted to assess the impact of selenium-enriched conditions on the genetic composition of the strains.

We have re-written the "Strains and Reagents"

We have rewritten the abstract to accurately reflect the experimental process.

We have provided the complete names of the abbreviations mentioned for the first time in the manuscript, including, MRS (De Man, Rogosa, and Sharpe), and Phosphate-Buffered Saline (PBS).

We have mentioned the name and model of the genomic DNA extraction kit used in the manuscript, which is the Tiangen Bacterial Genomic DNA Extraction Kits (Tiangen, Beijing, China).

We have added references for the scanning electron microscopy (SEM) analysis, Fourier transform infrared (FT-IR) analysis, and X-ray diffraction (XRD) analysis in the manuscript.

We have indicated that the number of viable bacteria was determined using the plate coating method to calculate the colony-forming units (CFU).

We have specified the statistical test used in the manuscript, which is the Tukey test.

Thank you for your review and suggestions. We believe that these modifications further enhance the quality and clarity of our manuscript.

4. Results

They were purchased not selected? In this study, six strains of bacteria were selected 

Figure 1. Which alphabet represent which bacteria? Explain it clearly.

Reconstruct the Phylogenetic tree and differentiate your strains (bold or colr).

Why the authors are in rush. They wrote a short caption for Figure 3. They must explain all the important information regarding the figure.

Response to comment 4:

Thank you for your feedback on our manuscript. We have addressed your comments as follows:

We apologize for the confusion. We have revised the sentence.

We have added a clear legend to Figure 1.

We have reconstructed the phylogenetic tree and highlighted our strains in bold.

We have revised the caption for Figure 3 to provide a more detailed explanation of the important information presented in the figure.

5. Discussion

I could not find any discussion because the authors simply collated information from the literature.

The order of the results must have been followed in the discussion.

Response to comment 5:

We apologize for the lack of discussion. We have revised the manuscript to provide a more comprehensive discussion of our results, following the order presented in the Results section.

6. Conclusion

All the findings were repeated in the conclusion. For instance, discussing 16Sr RNA is not necessary.

Throughout the research trial, six strains were examined; however, the author concluded that "strain LA 6076 was" chosen.

Rewrite it entirely, mentioning just the standout outcomes.

Response to comment 6:

Thank you for your valuable feedback, which has helped us improve the clarity and quality of our manuscript. We have rewritten the conclusion to focus on the standout outcomes of our study and avoid unnecessary repetition of information.

Once again, we appreciate your review and guidance, and we will continue to strive for your approval.

Best regards,

Tianli Yue

Reviewer 4 Report

On overall quality of the paper is good, however some issues require corrections or explanations.

Title: "prebiotic" is not the same as "probiotic"

Abbreviations should be defined not only in the Abstract, but in the main text as well. LA abrr. is not explained anywhere

Line 49: The sentence suggests that all LAB are probiotics, which is not true

Lines 73-74: latin names of the species shuld be written in italic; names LA 21805 and LA 6076 should be completed by latin names

Line 77: chemical form of selenium should be given

Lines 224-225: Had Authors expected that Se would change genetic identity of the strains?

Lines 256-261 and 262-266: the same information repeated

Lines 44, 220, 230: "Se" instead of "se"

Lines 51-53: plural form - "help", "reduce"

Line 60: excessive space

Line 61: "bioaccumulation and biotranformation capacity"

Line 70: missing space

Line 91: "biomass" rather than "body"

Line 112: for "species" both singural and plural forms are the same

Line 196: passive voice should be used

Lines 215-218: dots/semicolons missing

Line 281: of Se

Line 349: passive voice should be used

Author Response

Dear Reviewer:

Thank you for the comments concerning our manuscript entitled "Screening, characterization and probiotic properties of selenium-enriched lactic acid bacteria".

Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made correction which we hope meet with approval. Revised portion are marked in the paper. The main corrections in the paper and the responds to the reviewer’s comments are as flowing:

1. Title: "prebiotic" is not the same as "probiotic"

Response to comment 1: Thank you for your significant reminding. We have corrected the title to "Screening, characterization and probiotic properties of selenium-rich lactic acid bacteria."

2. Abbreviations should be defined not only in the Abstract, but in the main text as well. LA abrr. is not explained anywhere

Response to comment 2：We apologize for the oversight. We have provided an explanation for "LA" when it first appears in the abstract.

3. Line 49: The sentence suggests that all LAB are probiotics, which is not true

Response to comment 3：We apologize for any confusion caused by the sentence suggesting that all LAB are probiotics. We have rewritten and supplemented this section of the abstract.

4. Lines 73-74: latin names of the species shuld be written in italic; names LA 21805 and LA 6076 should be completed by latin names

Response to comment 4：We appreciate your feedback. We have italicized the Latin names of species and added the complete Latin names for LA 21805 and LA 6076.

5. Line 77: chemical form of selenium should be given

Response to comment 5：Thanks for your suggestion. We have added the chemical form of selenium.

6. Lines 224-225: Had Authors expected that Se would change genetic identity of the strains?

Response to comment 6：Thank you for your interest in this issue. In our study, we did not specifically anticipate that selenium would alter the genetic characteristics of strains. However, considering that selenium's interactions with biological systems are typically complex and multifaceted, we conducted 16S rRNA determinations to observe whether selenium had affected the genetic characteristics of the strains. We plan to further investigate this issue in our future research to gain a better understanding of selenium's role in microbial systems. Once again, we appreciate your valuable input.

7. Lines 256-261 and 262-266: the same information repeated

Response to comment 7：We apologize for the oversight and have eliminated the repeated information.

8. Lines 44, 220, 230: "Se" instead of "se"

Response to comment 8：We apologize for the error. We have corrected "se" to "Se" in the mentioned lines.

9. Lines 51-53: plural form - "help", "reduce"

Response to comment 9：Thank you for the correction. We have adjusted the plural forms.

10. Line 60: excessive space

Response to comment 10：We appreciate your observation. Excessive space has been removed from Line 60.

11. Line 61: "bioaccumulation and biotranformation capacity"

Response to comment 11：Thanks for your suggestion. We have updated the phrasing to "bioaccumulation and biotransformation capacity."

12. Line 70: missing space

Response to comment 12：We apologize for the missing space and have corrected it in Line 70.

13. Line 91: "biomass" rather than "body"

Response to comment 13：Thank you for pointing this out. We have replaced "body" with "biomass" in Line 91.

14. Line 112: for "species" both singural and plural forms are the same

Response to comment 14：Thank you very much for your suggestion. We have made the necessary adjustments for the term "species."

15. Line 196: passive voice should be used

Response to comment 15：We appreciate your suggestion. Passive voice has been used in Line 196.

16. Lines 215-218: dots/semicolons missing

Response to comment 16：Thank you for the observation. We have added the missing dots/semicolons in the specified lines.

17. Line 281: of Se

Response to comment 17：Thank you for your suggestions. We have revised "of Se" in Line 281 as suggested.

18. Line 349: passive voice should be used

Response to comment 18：Thank you very much for your suggestion. We have made revisions to this section.

Once again, we appreciate your review and guidance, and we will continue to strive for your approval.

Best regards,

Tianli Yue

Round 2

Reviewer 3 Report

I was rejected this manuscript due to significant flaws. The authors asserted that they enhanced its writing and corrected grammatical mistakes, but regrettably, I still identified them. I highlighted these issues. Moreover, the article contains numerous errors, and I provided recommendations to enhance its quality. Kindly address these concerns point by point. It could be published in the current issue of fermentation once address the concerns

Fully spell out the abbreviation the first time it is used. MRS?

How the same starin could be compared. " Lactobacillus plantarum 6076, Se-enriched Lactobacillus plantarum 6076 exhibited enhanced.

Lactobacillus plantarum 6076. No need to write it complete again and again. Replace. L. plantarum 6076

In the beginning the author generakizes and used LAB and later on, he focused on L. plantarum 6076. It created confusion.

Sentence #21 and 22 used twice LAB.

What the authors regions mean here "countries and regions"

Grammar mistakes, LAB play… LAB plays

The abstract doesn’t say anything about

Lactobacillus plantarum 21805, Lactobacillus paracasei 20241, Lactobacillus fermentum

21828, Lactobacillus casei 23185, Lactobacillus acidophilus 6064, Lactobacillus plantarum 6076

All strains were incubated in MRS. You mean grow in this medium. Incubation could be in incubator but this is a medium right

Activated mean here "and activated to the 3rd generation"

Minor editing of English language required

Author Response

Dear Editors and Reviewer,

Thank you for your thorough review and valuable feedback on our manuscript. We appreciate the time you've taken to highlight the areas that need improvement. We have addressed each of your concerns point by point and make the necessary revisions to enhance the quality of the article.

1. Reviewer Comment: Fully spell out the abbreviation the first time it is used. MRS?

Response: We apologize for our oversight. "MRS" (de Man, Rogosa, and Sharpe agar) has been fully spelled out. Thank you for bringing this to our attention.

2. Reviewer Comment: How the same strain could be compared. " Lactobacillus plantarum 6076, Se-enriched Lactobacillus plantarum 6076 exhibited enhanced.

Response: Thank you for highlighting this potential confusion. We have made modifications to the article.

3. Reviewer Comment: Lactobacillus plantarum 6076. No need to write it complete again and again. Replace. L. plantarum 6076,

Response: We apologize for any confusion caused by the repeated use of the complete term. The abbreviation "L. plantarum 6076" has been used to replace the complete term "Lactobacillus plantarum 6076" to avoid repetitive usage.

4. Reviewer Comment: In the beginning, the author generalizes and uses LAB, and later on, he focuses on L. plantarum 6076. It created confusion.

Response: We appreciate your feedback on this matter. The manuscript has been revised to ensure consistency in terminology throughout, with the use of "LAB".

5. Reviewer Comment: Sentence #21 and 22 used twice LAB.

Response: Thank you for bringing this to our attention. We have addressed this repetition.

6. Reviewer Comment: What the authors regions mean here "countries and regions"?

Response: We apologize for the confusion. We have replaced and described the term to eliminate ambiguity.

7. Reviewer Comment: Grammar mistakes, LAB play… LAB plays

Response: We appreciate your feedback on grammar. Grammar mistakes, such as "LAB play" have been carefully reviewed and corrected to "LAB plays," as you've pointed out.

8. Reviewer Comment: The abstract doesn’t say anything about Lactobacillus plantarum 21805, Lactobacillus paracasei 20241, Lactobacillus fermentum 21828, Lactobacillus casei 23185, Lactobacillus acidophilus 6064, Lactobacillus plantarum 6076.

Response: We apologize for any oversight in the initial abstract. We have updated the abstract to include relevant information including Lactobacillus plantarum 21805, Lactobacillus paracasei 20241, Lactobacillus fermentum 21828, Lactobacillus casei 23185, Lactobacillus acidophilus 6064, and Lactobacillus plantarum 6076.

9. Reviewer Comment: "All strains were incubated in MRS." You mean grow in this medium. Incubation could be in incubator but this is a medium right

Response: Thank you for pointing out the need for clarity in this regard. We have modified the appropriate terminology.

10. Reviewer Comment: Activated mean here "and activated to the 3rd generation"

Response: We appreciate your feedback. We have revised the text to provide a clear explanation in the context for better understanding.

We are committed to addressing these concerns and enhancing the manuscript's quality to meet the standards for publication in the issue of Fermentation. We will make the revisions promptly and resubmit the manuscript for your review.

Thank you once again for your valuable feedback.

Sincerely,

Tianli Yue