Applications of Food-Associated Lactobacillaceae in Fermented Foods, Health, and Emerging Biotechnologies

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This article provides a comprehensive overview of lactic acid bacteria (LAB) applications across multiple fields, including human health, food fermentation, agriculture, livestock, and vaccination. While the manuscript is generally well-structured, it would be better to incorporate more recent advancements in the laboratory study of LAB, particularly in the areas of food and health.

Main concerns:

1. Title Scope: The current title, which emphasizes LAB applications in fermented foods and beverages, does not fully reflect the broader scope of the article's content.

2. The discussion on fermented foods and beverages should include the specific roles of LAB in flavor profiles and bioactive compounds in fermentation products.

3. Recent advances on the contribution of LAB-derived extracellular vesicles to gut health should be integrated into the manuscript.

Minor point: 

1. In the manuscript, Figure 1, 3 and 4 are actually tables. 

Author Response

We sincerely thank the reviewer for their careful evaluation of our manuscript and for the constructive comments and suggestions, which have helped us improve the quality, clarity, and scope of the article. We have addressed each comment in detail below. All changes have been incorporated into the revised manuscript.

General

Reviewer comment:

This article provides a comprehensive overview of lactic acid bacteria (LAB) applications across multiple fields, including human health, food fermentation, agriculture, livestock, and vaccination. While the manuscript is generally well-structured, it would be better to incorporate more recent advancements in the laboratory study of LAB, particularly in the areas of food and health.

Response:
We thank the reviewer for this positive assessment. In response, we have incorporated more recent advancements in the laboratory study of lactic acid bacteria, particularly related to food and health applications. Notably, we have updated the manuscript to reflect Zheng et al.’s reclassification of the Lactobacillaceae, ensuring that current taxonomic and functional insights are accurately represented throughout the text.

>Main Concerns

1. Title Scope

Reviewer comment:
The current title, which emphasizes LAB applications in fermented foods and beverages, does not fully reflect the broader scope of the article's content.

Response:
We agree with the reviewer’s observation and have revised the title to better reflect the comprehensive scope of the manuscript. The new title is:

“Applications of Food-Associated Lactobacillaceae in Fermented Foods, Health, and Emerging Biotechnologies.”

This revised title more accurately captures the breadth of applications discussed in the manuscript.

2. Roles of LAB in Flavor Profiles and Bioactive Compounds; extracellular vesicles

Reviewer comment:
The discussion on fermented foods and beverages should include the specific roles of LAB in flavor profiles and bioactive compounds in fermentation products. Recent advances on the contribution of LAB-derived extracellular vesicles to gut health should be integrated into the manuscript.

Response:
We appreciate this suggestion and have expanded the discussion accordingly. A new subsection (Section 3.3: Roles of Food-Associated Lactobacillaceae in Flavor Development and Bioactive Compound Formation) has been added to detail the metabolic contributions of LAB to flavor development, aroma compounds, and the generation of health-relevant bioactive molecules during fermentation. We have also added a dedicated subsection (Section 3.5: Extracellular Vesicles Derived from Food-Associated Lactobacillaceae), which discusses recent findings on LAB-derived extracellular vesicles and their roles in host–microbe interactions, gut health, and immunomodulation.

Minor Point

1. Figures vs. Tables

Reviewer comment:
In the manuscript, Figure 1, 3, and 4 are actually tables.

Response:
We agree and have corrected this issue. Figures 1, 3, and 4 have been relabeled appropriately as tables.

Reviewer 2 Report

Comments and Suggestions for Authors

Paper: “Application of Lactobacillus in Fermented Food and Beverages”

by Pathan et al., Fermentation

The subject of this review (Lactobacillus in fermented foods) has already been extensively covered in the literature, and this manuscript provides neither original contributions nor innovative summaries. The review appears largely compilatory, lacking a critical or genuinely innovative approach. The figures and tables are mostly descriptive summaries and add no interpretative or conceptual value. Some of the most notable errors authors treats Lactobacillus as a single genus, ignoring the 2020 taxonomic revision that split it into 25 genera. This is a serious error for a review published in 2025. The abstract is generic and fails to highlight a knowledge gap or a clear objective. The introduction is verbose and repetitive, lacking a critical framework. Section progression is incoherent: the text jumps from probiotics to fermentation, then to safety, then to industrial applications, without logical flow and many paragraphs are descriptive rather than analytical. References are mixed: some outdated, others not directly relevant. Key recent works (2020–2025) on taxonomy, functional genomics, and biotechnological applications of the new genera are missing. Certain data (e.g., probiotic market size, clinical trial numbers) are reported without proper contextualization or primary sources. The language is often colloquial and unsuitable for a scientific review and there are frequent repetition of concepts already stated. The conclusions are generic and fail to provide a critical synthesis or concrete perspectives. No discussion of literature limitations or methodological challenges. 

In summary, this review contains serious taxonomic and nomenclatural errors, lacks originality and a critical approach. The structure is confusing and the language is unscientific. Therefore, the manuscript does not meet the minimum standards required for a scientific review in an international journal.

Comments on the Quality of English Language

The language is often colloquial and unsuitable for a scientific review.

Author Response

We thank the reviewer for their thorough assessment of our manuscript. We take these comments seriously and have substantially revised the manuscript to address concerns related to scope, structure, taxonomy, critical analysis, and scientific rigor.

First, we would like to clarify the issue of taxonomy. The 2020 reclassification of the former Lactobacillus genus, as proposed by Zheng et al., was cited and acknowledged in the original submission. However, in the revision, we have emphasized this update further, recognizing that in many sections of the initial draft, references to “Lactobacillus” were not consistently updated to reflect the 25 new genera. We also note that although Zheng’s reclassification occurred in 2020, some post 2020 articles, even in MDPI, still refer interchangeably to Lactobacillaceae using older terminology. In the revised manuscript, we have rigorously applied the current nomenclature across all sections, tables, and figures, ensuring that species are correctly attributed to their respective genera (e.g., Lactiplantibacillus, Limosilactobacillus, Levilactobacillus, Lacticaseibacillus), and we have highlighted functional and health-relevant differences to provide a more accurate and nuanced discussion.

Regarding originality and critical synthesis, we have restructured the manuscript to move beyond a descriptive compilation. The review now explicitly frames post-2020 advances in functional genomics, metabolomics, extracellular vesicles (subsection 3.5) , and emerging biotechnological applications, emphasizing the importance of strain–matrix interactions and postbiotic signaling. This conceptual lens helps interpret health effects, fermentation outcomes, and industrial applications, allowing readers to understand these microorganisms within complex food matrices rather than as interchangeable probiotics.

We have also improved the logical flow, moving systematically from taxonomy and health effects to fermentation mechanisms, safety considerations, industrial applications, and future perspectives. Figures and tables have been revised to provide interpretative insight, highlighting trends, mechanisms, and strain-specific effects. References have been updated to include key studies from 2020–2025.

Finally, the manuscript’s language has been polished for formal scientific style, redundancy has been minimized, and conclusions now provide a critical synthesis with concrete perspectives on research gaps, methodological challenges, and translational potential. We maintain that this is a review rather than an original experimental study. We believe these revisions adequately address the reviewer’s concerns, enhancing taxonomic accuracy, analytical depth, and overall coherence, while maintaining the review’s relevance and contribution to the current understanding of food-associated Lactobacillaceae.

Reviewer 3 Report

Comments and Suggestions for Authors

I  think this review give us a very good summary about the applications of  Lactobacillus in Fermented Foods and Beverages，Agricultural and Livestock，Safety， Vaccination，  Challenges and Future Perspectives, I think  this manuscript  can be published after the authors make a minor revise , My revision opinions are as follows:

 For  Therapeutic intervention, besides  Antihypertensives, Lipid lowering agents, Infection prevention, some  Lactobacillus also  have  ability for gout treatment，diabetes  lowering，liver protecting  etc.  it is better if the author can supplement  addition relevant literature 

 

Author Response

We thank the reviewer for their positive evaluation of our manuscript and for recognizing its comprehensive coverage of Lactobacillaceae applications across fermented foods and beverages, agriculture and livestock, safety, vaccination, and future perspectives. We appreciate the constructive suggestion regarding therapeutic interventions. In response, we have expanded the relevant sections in Table 1 and Section 4.2 to include additional biochemical mechanisms and evidence on the roles of specific Lactobacillaceae strains in gout management, glycemic control in diabetes, and hepatoprotective effects. Appropriate recent literature has been added to support these functions, and the discussion has been integrated within a strain-specific and mechanistic framework. These additions strengthen the clinical and translational relevance of the review while remaining aligned with its scope as a narrative synthesis.

Reviewer 4 Report

Comments and Suggestions for Authors

General comments:

Lactobacillus is subdivided into new genera as of 2020. Authors should use new nomenclature, or simply use "members of family Lactobacillaceae" 

Introduction is very scarce and somewhat redundant. Different roles that lacotabcilli may play should be explained in more detail, especially their role in the fermentation processes.

The chapter addressing Lactobacillus and probiotic properties is not adequately connected to the topic of the manuscript. It does not explain how lactobacilli originating from foods and beverages exert probiotic effects within those matrices. Moreover, lactobacilli are frequently administered as probiotics via pharmacological formulations, and is therefore not relevant in this context. Consequently, the chapter should be revised to include only studies in which lactobacilli were deliberately incorporated into food and beverage matrices.

Chapter 4 is not connected in any way with the title of the manuscript

Genus should be written in italic

Lines 30-32. This sentence is somewhat confusing and redundant. Development of sensory properties is part of the fermentation process, and it should not be explained separately. Also, not all lactobacilli are probiotics, this is a property of the specific strains, rather than all lactobacilli. Authors are advised to be more precise.

lines 34-36: as probiotic properties are tied to a specific strains, this statement is too general. Most of Lacobacilli are not probiotics - therefore, it should be clearly stated that "some strains, members of Lactobacillaceae family are considered as probiotics" or  the strains (for example LL. acidophilus LA5) should be written. 

Line 46: References are missing

line 56: Genus is not a "probiotic group" but a taxonomic category. Therefore, this sentence should be written in a different manner.

Line 73: If the authors write about the strains, then the complete names of the strains should be written

Lines 78-83: the connection between antibiotic usage and bacteriocin application is somewhat unclear. Bacteriocins are currently used for food preservation properties, and not as therapeutics

Line 94: references should be given in parenthesis as numbers

Lines 113-116: There is only one research mentioned in the text, but two references are cited. 

Lines 116-123: references should be cited only by numbers, and not the names.

lines 125-129: citation for the studies is missing

Lines 134-139: the original research should be cited here, and not the review 

Lines 176-190: new taxonomy should be used

Author Response

We thank the reviewer for the detailed and constructive comments, which have helped us significantly improve the accuracy, clarity, and focus of the manuscript. We have carefully revised the manuscript to address the chief concerns raised.

Regarding taxonomy and nomenclature, we emphasized the 2020 reclassification of the former Lactobacillus genus into multiple genera within the family Lactobacillaceae by moving it to the abstract. More importantly, in the revised manuscript, we now consistently use the updated nomenclature or the collective term members of the family Lactobacillaceae where appropriate. All genus and species names have been formatted in italics throughout the manuscript. We note lamentably that some inconsistencies in italics and genus formatting were introduced during conversion to the MDPI format; this formatting error has now been corrected.

The Introduction has been expanded and streamlined to reduce redundancy and to better articulate the diverse roles of Lactobacillaceae in fermentation processes, including their contributions to acidification, flavor and texture development, microbial stability, and safety. We now explicitly frame sensory development as an intrinsic component of fermentation rather than a separate process, and we clarify that probiotic properties are strain-specific rather than genus-wide.

The section on probiotic properties has been substantially revised to improve its relevance to the manuscript’s focus. We enhanced discussion to studies in which Lactobacillaceae strains were deliberately incorporated into food and beverage matrices and discuss how probiotic effects are exerted within those matrices. References to pharmacological or capsule-based probiotic administration have been corrected.

The manuscript title has been addended and is now inclusive of Chapter 4 which we maintain is a significant component of this review. 

Throughout the manuscript, we have improved precision by explicitly distinguishing strains from genera, providing full strain designations where applicable, and revising language that incorrectly referred to genera as “probiotic groups” (the corrected version now moved to abstract). Statements implying that all lactobacillaceae are probiotics have been corrected to reflect strain-specific evidence (e.g., Lacticaseibacillus rhamnosus GG, Lactiplantibacillus acidophilus LA-5).

Major citation issues noted by the reviewer have been addressed: missing references have been added; citations are now consistently formatted as numerical references in parentheses; duplicate or mismatched citations have been corrected; and original research articles have replaced reviews where appropriate. We also clarified the discussion on bacteriocins to reflect their established role in food preservation rather than therapeutic use.

Finally, the revised manuscript consistently applies the updated taxonomy in all relevant sections, including Lines 176–190, ensuring taxonomic accuracy and compliance with current standards.

We believe these revisions substantially improve the manuscript’s scientific rigor, coherence, and relevance to fermented food and beverage research, and we sincerely thank the reviewer for their valuable guidance.

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Compared with the previous version, the revised manuscript is significantly clearer and more logically coherent. Recent advances have been incorporated, enabling a more thorough discussion across all sections of the manuscript. The authors have also addressed the reviewers' questions and suggestions. In my view, the paper is now suitable for publication.

Author Response

We sincerely thank the reviewer for their detailed input throughout the review process.

Reviewer 2 Report

Comments and Suggestions for Authors

The authors have significantly improved and reorganized the work. I have no further suggestions.

Author Response

We sincerely thank the reviewer for their input throughout the review process.

Reviewer 4 Report

Comments and Suggestions for Authors

-

Author Response

We sincerely thank the reviewer.