Exploring the Intriguing World of Fungal Diversity in the Oral Cavities of a Native Community in Siltepec, Chiapas, Mexico
and Victor E. Aguirre-Arzola 2,*Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsManuscript of considerable interest for the dental sector, requires a major revision before considering a possible publication.
Abstract: not very attractive, highlight the results obtained more.
Few keywords: add specific ones.
Introduction: very sparse, add all the specifications on the oral microbiota and how it changes with respect to the various pathologies and all the minimally invasive systems to maintain a state of homeostasis over time (Scribante et al).
Materials and methods; how was the sample size calculated? The reference is missing.
Very confusing results, reorganize the tables to make them more fluent for the reader, figure 4 overlies table two which is not readable, and in turn figure 4 is at low resolution.
Discussion: add proactive action through natural substances, as future objectives (Butera et al).
Conclusions: modify them based on the changes in the text.
Bibliography: add references requested.
Author Response
Dear Reviewer
We sincerely thank you for your thoughtful and constructive feedback on our manuscript titled “Exploring the Intriguing World of Fungal Diversity in the Oral Cavity of a Native Community in Silpetec, Chiapas, Mexico.” We have carefully reviewed all your suggestions and made significant revisions to the manuscript to improve its clarity, depth, and overall contribution to the field of oral microbiology and fungal diversity. Below is a detailed response to each of your comments:
- Abstract: not very attractive, highlight the results obtained more.
We have revised the abstract to emphasize the key results of our study, particularly the significant findings regarding the relationship between filamentous fungi and dental caries. The revised abstract is now more engaging and focuses on the protective role of fungi, highlighting the influence of a maize-based diet and the consumption of fermented beverages like pozol on the oral microbiome. We believe these changes better capture the significance of our research.
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- Few keywords: add specific ones.
In response to your suggestion, we have expanded the list of keywords to include more specific terms such as Cladosporium , oral microbiome, maize-based diet, fermented beverages, microbial competition, and indigenous communities. This provides a clearer representation of the study’s focus and will make it more accessible to relevant audiences.
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- Introduction: very sparse, add all the specifications on the oral microbiota and how it changes with respect to the various pathologies and all the minimally invasive systems to maintain a state of homeostasis over time (Scribante et al).
We have expanded the introduction to include detailed information about the oral microbiota, how it shifts in response to various pathologies, and the role of minimally invasive strategies in maintaining oral homeostasis over time. In particular, we referenced the work of Scribante et al. (2022) to provide context for how natural substances, such as probiotics and plant-based antimicrobials, can play a role in oral health. This addition enhances the introduction and provides a strong foundation for our research.
- Materials and Methods: how was the sample size calculated? The reference is missing.
We have now included the complete results and expanded the sample size to strengthen the statistical robustness of our findings. Our sampling approach follows standard techniques commonly applied in rural health studies, taking into account the unique characteristics of the indigenous population in Silpetec, Chiapas. The expanded sample provides a more comprehensive representation of the population, and we have also added a reference to support the methodology. This adjustment ensures that our study’s conclusions are more representative and statistically sound.
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- Very confusing results; reorganize the tables to make them more fluent for the reader, figure 4 overlies table two which is not readable, and in turn, figure 4 is at low resolution.
We have reorganized the results section to improve clarity and fluency for the reader. The tables have been restructured to present the data in a more intuitive way, and Figure 4 has been replaced with a high-resolution version. The layout has been adjusted to avoid overlapping with Table 2, ensuring that both the figure and table are fully legible. These changes greatly enhance the readability of the results section.
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- Discussion: add proactive action through natural substances, as future objectives (Butera et al).
In the revised discussion, we have included a proactive perspective on the use of natural substances, particularly plant-based antimicrobials and probiotics , as future objectives for promoting oral health. Drawing on the work of Butera et al. , we discuss how these natural interventions can be used to maintain oral homeostasis and prevent dental caries, particularly in rural and indigenous populations where access to conventional dental care may be limited. We believe this addition aligns with current research trends and strengthens the future direction of the study.
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- Conclusions: modify them based on the changes in the text.
We have revised the conclusions to reflect the significant revisions made throughout the manuscript. The conclusion now emphasizes the protective role of filamentous fungi, the influence of traditional maize-based diets on the oral microbiome, and the potential application of natural substances for preventive oral health strategies.
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- Bibliography: add references requested.
The bibliography has been updated to include the necessary references, including Scribante et al. (2022) and Butera et al. , to support the expanded discussion on oral microbiota and the use of natural interventions for maintaining oral health. All requested references have been added, ensuring the manuscript is well-supported by current literature.
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Additional Notes on Study Limitations and Sample Size:
While we have expanded the sample size to provide more comprehensive results, it is important to note that the unique geographical and cultural characteristics of the indigenous communities in Silpetec, Chiapas, make this study especially valuable despite the inherent challenges in accessing a larger population. We believe that focusing on this isolated and understudied population adds a unique dimension to the research, contributing valuable insights into the relationship between traditional diets, fungi, and oral health. This focus on an isolated population also provides an opportunity for future longitudinal studies to explore these dynamics further.
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We greatly appreciate your insightful feedback, which has significantly improved the quality of our manuscript. We are confident that the revisions we have made address your concerns and enhance the overall contribution of the study to the field of oral microbiology and indigenous health research. Thank you once again for your time and thoughtful suggestions. We look forward to your further feedback.
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Sincerely,
Victor E. Aguirre-Arzola
Facultad de agronomía, UANL
victor.aguirrearz@uanl.edu.mx
Reviewer 2 Report
Comments and Suggestions for AuthorsDear Authors,
I am providing information about irregularities in the research and the article in which the description of the research results was included.
Incorrect methodology: too small a sample size – 37 people, predominance of women – 21 women and 9 men, too large an age range – from 1 to 68 years, no information on the selection of patients – native patients. It seems that there are more indigenous people living in Silpetec, Chiapas, Mexico than the size of the sample size.
The introduction text lacks reference number 2, this reference was placed later in the article.
Reference number 6 and 17 are the same article.
The relationship between the type of advancement of detected dental caries and the number of detected filamentous fungi was not statistically analyzed.
The discussion is unconvincing – no references to other mycological works concerning the oral cavity. The discussion referred only to 4 items of literature, which are older than 5 years. Discussion to be supplemented.
The article should be corrected and supplemented.
Author Response
Dear Reviewer,
Thank you for your thoughtful and detailed feedback on our manuscript, “Exploring the Intriguing World of Fungal Diversity in the Oral Cavity of a Native Community in Silpetec, Chiapas, Mexico.” We have carefully considered your comments and made substantial revisions to address each of your concerns. Below, we provide a point-by-point response to the issues you raised, including the updates and justifications for our approach.
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- Incorrect methodology: too small a sample size, predominance of women, wide age range, and no clear selection criteria for native patients.
We understand your concern regarding the original sample size and demographics. We are pleased to inform you that the study now includes the complete dataset of over 140 participants , significantly expanding the sample size from the originally reported 37 individuals. This study was conducted as part of a free health campaign in the indigenous communities of Silpetec, Chiapas, which provided much-needed oral health services to the population.
While there is a predominance of women (99 female participants), this reflects the gender distribution of those who attended the campaign. The age range (15 to 86 years) was also adjusted to ensure that a broad representation of oral health conditions could be analyzed across the community.
We have now clarified the selection criteria in the Materials and Methods section, confirming that only participants who self-identified as indigenous were included in the study. With this larger and more comprehensive sample, the statistical power of our findings has been improved, and the results are now more representative of the broader population of Silpetec.
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- The introduction text lacks reference number 2; this reference was placed later in the article.
We apologize for this oversight and have corrected it. Reference number 2 is now properly cited within the introduction where it is first mentioned, ensuring continuity and accuracy in the citation flow throughout the manuscript.
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- Reference number 6 and 17 are the same article.
We appreciate you bringing this to our attention. The duplicated reference has been removed, and the bibliography has been updated to ensure that each reference is unique. All references now correctly correspond to distinct sources.
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- The relationship between the type of advancement of detected dental caries and the number of detected filamentous fungi was not statistically analyzed.
Thank you for highlighting this important aspect. In response, we have conducted additional statistical analyses to assess the relationship between the severity of dental caries and the presence of filamentous fungi. The results of this analysis are now included in the revised Results and Discussion sections, providing a clearer understanding of the correlations between caries progression and fungal presence.
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- The discussion is unconvincing – no references to other mycological works concerning the oral cavity, and it relies on outdated sources.
We have significantly expanded the Discussion section to address this concern. Additional references to recent mycological studies focusing on the oral cavity have been included, and the literature now reflects more contemporary research published within the last five years. This includes new references on the interaction between fungi and oral diseases, as well as works related to the oral microbiome. We believe these additions strengthen the scientific foundation of the discussion and provide a more convincing narrative.
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- The article should be corrected and supplemented.
In addition to the points mentioned above, we have made comprehensive revisions throughout the manuscript to correct and supplement the article. The Results section now includes additional analyses, the tables and figures have been adjusted for clarity and readability, and the References section has been thoroughly reviewed to ensure all citations are accurate and up-to-date. These changes address your concerns and enhance the overall quality of the manuscript.
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We are grateful for your careful review and appreciate the opportunity to improve our manuscript. The revisions we have made, including the expanded sample size and updated analyses, have strengthened the scientific rigor of the study. We hope these changes meet your expectations and we look forward to your further feedback.
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Sincerely,
Victor E. Aguirre-Arzola
Facultad de agronomía, UANL
victor.aguirrearz@uanl.edu.mx
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Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript titled "Exploring the Intriguing World of Fungal Diversity in the Oral Cavity of a Native Community in Silpetec, Chiapas, Mexico" presents a significant contribution to the study of the oral mycobiome, particularly within a specific and underexplored population. The research focuses on a rural community with unique dietary and environmental conditions, which provides a novel perspective on the relationship between these factors and the composition of the oral fungal microbiota. This is a compelling aspect of the study that highlights the potential for new insights into how diet and environment influence microbial communities in the human body.
The study employs a robust methodological approach, combining both morphological and genetic characterization of fungal isolates. This dual approach is a strong point of the manuscript, as it allows for a more comprehensive identification of the fungi present in the oral cavity of the study participants. The use of polymerase chain reaction (PCR) and 28S rRNA sequencing provides a reliable means of species identification, adding credibility to the findings. The study's focus on both culturable and non-culturable fungi further strengthens its contribution to the understanding of the oral mycobiome, as it acknowledges the complexity of fungal communities beyond what can be observed through traditional culturing techniques.
However, despite these strengths, the manuscript requires significant revisions to improve its clarity, structure, and overall impact. The introduction, while informative, could benefit from a more focused narrative that clearly defines the research question and the specific contributions this study intends to make. Currently, the introduction tends to be somewhat broad, which may dilute the study's unique angle. The authors should aim to sharpen the research focus, making it evident how this study addresses gaps in the existing literature.
The methods section, though detailed, includes redundancies that could be streamlined to enhance readability. Descriptions of laboratory procedures should be concise and focused on aspects that are critical for reproducibility. Some sections, particularly those detailing the genetic characterization, are overly technical and could benefit from further explanation or simplification to ensure they are accessible to a broader scientific audience.
The results are presented with a high level of detail, which, while thorough, can lead to repetition and may overwhelm the reader. The authors should consider summarizing some of the repetitive information, perhaps through the use of tables or figures, to make the findings more digestible. Additionally, the discussion of the results could be expanded to offer deeper insights into the implications of the findings. The current discussion sometimes merely restates the results without fully exploring their significance or how they advance our understanding of the oral mycobiome.
The literature review is generally well-cited, but there are instances where more recent or relevant studies could be included. The inclusion of contemporary research, particularly studies that directly relate to the oral mycobiome, would provide a stronger contextual foundation for the study's findings. The authors should ensure that their discussion is anchored in the most current understanding of the field.
Another area that requires attention is the language and style of the manuscript. There are several instances of awkward phrasing and grammatical errors that detract from the readability of the text. The manuscript would benefit from a thorough review by a native English speaker or a professional editor to improve its fluency and clarity. In addition, some statements in the manuscript are vague or unclear, which could lead to misinterpretation by the reader. For instance, the discussion on the influence of diet on the oral microbiome is intriguing, but the lack of specific explanations or hypotheses makes it less compelling. The authors should strive for greater precision in their language to convey their findings and interpretations more effectively.
The conclusions drawn from the study are interesting but seem somewhat speculative in light of the data presented. The authors should be cautious in making broad claims and ensure that their conclusions are well-supported by the evidence. It would also be beneficial to suggest directions for future research, particularly studies that could further explore the relationship between diet, geography, and the oral microbiome.
In summary, while the manuscript provides valuable contributions to the study of the oral mycobiome, particularly in a unique population, it requires substantial revisions to enhance its clarity, coherence, and impact. By addressing the issues related to the focus of the research question, the presentation of methods and results, the depth of the discussion, and the language used, the authors can significantly improve the manuscript. With these improvements, the study has the potential to make a meaningful contribution to the field of microbial ecology and our understanding of the factors that shape the human oral microbiome.
Author Response
Dear Reviewer,
Thank you for your thoughtful and detailed feedback on our manuscript, “Exploring the Intriguing World of Fungal Diversity in the Oral Cavity of a Native Community in Silpetec, Chiapas, Mexico.” We have carefully considered your comments and made substantial revisions to address each of your concerns. Below, we provide a point-by-point response to the issues you raised, including the updates and justifications for our approach.
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1.0 introduction We appreciate your suggestion and have revised the Introduction to provide a more focused and clear narrative. We have sharpened the research question and clearly articulated the specific contributions of this study. In particular, we now emph: size how our work addresses key gaps in the literature related to the interaction between diet, environment, and the oral mycobiome in indigenous communities. This revision clarifies the novel aspect of the research and situates it within the broader context of current oral microbiome studies.
2.0 Methods: Redundancies and overly technical sections.
In response to your feedback, we have streamlined the Methods section to remove redundancies and enhance readability. Descriptions of laboratory procedures, particularly those concerning the genetic characterization of fungal isolates, have been revised to be more concise and focused on critical details necessary for reproducibility. We have also simplified the technical aspects of the genetic characterization, ensuring that the section is accessible to a broader scientific audience without sacrificing scientific rigor.
We understand your concern regarding the original sample size and demographics. We are pleased to inform you that the study now includes the complete dataset of over 140 participants , significantly expanding the sample size from the originally reported 37 individuals. This study was conducted as part of a free health campaign in the indigenous communities of Silpetec, Chiapas, which provided much-needed oral health services to the population.
While there is a predominance of women (99 female participants), this reflects the gender distribution of those who attended the campaign. The age range (15 to 86 years) was also adjusted to ensure that a broad representation of oral health conditions could be analyzed across the community.
We have now clarified the selection criteria in the Materials and Methods section, confirming that only participants who self-identified as indigenous were included in the study. With this larger and more comprehensive sample, the statistical power of our findings has been improved, and the results are now more representative of the broader population of Silpetec.
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3.0 Results and Statistical Analysis: To enhance the scientific robustness of the study, we have now included data from over 140 participants, strengthening the validity of our conclusions. We used Student's t-test and Mann-Whitney U test to compare caries prevalence between patients with and without filamentous fungi, providing greater scientific rigor and solidifying the reliability of our findings. These statistical tests demonstrated significant differences, adding substantial support to our hypothesis regarding the relationship between fungal presence and oral health outcomes. Additionally, we have summarized some of the repetitive information using tables and figures, making the findings more accessible and digestible for readers.
We have carefully revised the Results section to reduce repetition and improve readability. Repetitive information has been summarized, and we have utilized tables and figures more effectively to present the findings in a clearer and more digestible format. Additionally, we have expanded the Discussion to delve deeper into the implications of our findings. Rather than merely restating the results, we now explore their significance in relation to the oral mycobiome, highlighting how our study advances the understanding of fungal communities in the
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- Literature review: Include more recent and relevant studies.
We have updated the Literature Review to include more recent and relevant studies, particularly focusing on the oral mycobiome and fungal diversity in the oral cavity. Several contemporary studies have been cited, providing a stronger contextual foundation for the study’s findings. This revision ensures that the discussion is anchored in the most current understanding of the field and supports our interpretations of the results.
presence.
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- Language and style: Improve fluency, clarity, and precision.
We recognize the importance of clear and precise language, and we have addressed the grammatical errors and awkward phrasing identified in the manuscript. The manuscript has undergone a thorough review by a professional editor to ensure fluency and readability. We have also refined our language to ensure that the statements are specific and clear, particularly in the discussion of the influence of diet on the oral microbiome. The revised manuscript now conveys the findings and interpretations more effectively, minimizing the risk of misinterpretation
We have significantly expanded the Discussion section to address this concern. Additional references to recent mycological studies focusing on the oral cavity have been included, and the literature now reflects more contemporary research published within the last five years. This includes new references on the interaction between fungi and oral diseases, as well as works related to the oral microbiome. We believe these additions strengthen the scientific foundation of the discussion and provide a more convincing narrative.
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- The article should be corrected and supplemented.
In addition to the points mentioned above, we have made comprehensive revisions throughout the manuscript to correct and supplement the article. The Results section now includes additional analyses, the tables and figures have been adjusted for clarity and readability, and the References section has been thoroughly reviewed to ensure all citations are accurate and up-to-date. These changes address your concerns and enhance the overall quality of the manuscript.
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We are grateful for your careful review and appreciate the opportunity to improve our manuscript. The revisions we have made, including the expanded sample size and updated analyses, have strengthened the scientific rigor of the study. We hope these changes meet your expectations and we look forward to your further feedback.
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Here’s a carefully constructed response letter addressing the feedback provided by this reviewer. The response aims to acknowledge the strengths and weaknesses pointed out, and details how the manuscript has been revised for improvement:
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- Conclusions: Be cautious with broad claims and suggest future research directions.
We agree that caution is necessary when drawing conclusions based on the presented data. As such, we have revised the Conclusions to ensure that they are well-supported by the evidence provided in the manuscript. Additionally, we have included suggestions for future research, particularly in exploring the long-term effects of dietary patterns, geographical isolation, and their influence on the oral microbiome. These additions provide a clearer roadmap for further studies in this area.
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In summary, we have made substantial revisions to the manuscript, addressing the issues you raised regarding the focus of the research question, the presentation of methods and results, the depth of the discussion, and the overall clarity and language used. We believe these changes significantly improve the clarity, coherence, and impact of the manuscript. We are confident that these revisions have strengthened the manuscript and look forward to your further feedback.
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Sincerely,
Victor E. Aguirre-Arzola
Facultad de agronomía, UANL
victor.aguirrearz@uanl.edu.mx
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Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript has been almost revised in its entirety, references for future objectives in the discussion are missing as already suggested in the first round of revision (Butera A. et al)
Author Response
Dear Reviewer,
We sincerely appreciate your continued feedback on our manuscript. We have carefully considered your comments regarding the inclusion of references for future objectives in the discussion, particularly as suggested in the first round of revision, and we acknowledge the importance of citing relevant studies such as Butera et al. (2021).
In response to your suggestion, we have revised the discussion section to include appropriate references that align with the future research objectives of this study. Specifically, we have integrated the work of Butera et al., which provides a valuable perspective on the role of oral fungi in both health and disease contexts. Their study’s insights into the interaction between Candida, Aspergillus, and other fungal genera with oral health have helped strengthen our discussion on how local diets and environmental factors may influence fungal colonization and oral health outcomes in indigenous populations.
We believe that this revision addresses your concern and enriches the discussion with a forward-looking perspective, grounded in current research. We hope that this updated manuscript now meets your expectations and we thank you again for your insightful recommendations.
Sincerely,
Victor E. Aguirre-Arzola
vistor.aguirrearz@uanl.edu.mx
