Colonic Dysbiosis Is Associated with Gastrointestinal Disorders More than Mother–Child Epidemiological Traits in Mexican Children with Autism Spectrum Disorders
, Abraham Wall-Medrano 1, Arnulfo Ramos-Jiménez 1, Aarón F. González-Córdova 2, Florinda Jiménez-Vega 1,* and Rocio Campos-Vega 3,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe presented study describes the epidemiological characteristics and risk factors associated with gastrointestinal disorders in a group of children and adolescents with ASD residing in Mexico. The interesting aspect of this study is the in-depth analysis of these disorders in the early stages of development, in relation to intrauterine growth, cesarean delivery, and type of feeding. Literature has already described that these risk factors are associated with changes in the commensal microbiota and deviations in colonization patterns during early and late infancy. Therefore, identifying biomarkers that could be used in diagnostics is crucial to aid in the diagnosis and management of gastrointestinal complications. As mentioned by the authors, this study has certain limitations, such as the lack of more advanced sequencing techniques (NGS), which would provide a more comprehensive view of microbiota composition. Additionally, the dataset should be expanded to obtain more solid and robust data, especially in relation to complications. It would also be important to consider the presence of food selectivity and the type of diet followed by the patients and their families, as gastrointestinal disorders are also linked to nutrition.
Below are some points that should be revised and require further clarification.
- Line 19: It might be clearer to use the term “gut microbiota (GM)” instead of colonic-fecal microbiota. To be revised throughout the rest of the manuscript.
- Line 20-21: "A cases FGIDs (+) and controls FGIDs (-)” is not very clear; you could write “a study of FGIDs (+) cases and FGIDs (-) controls” for a clearer formulation.
- Line 22: "Distal gastrointestinal pathologies” could simply say “Gastrointestinal disorders” to improve the flow of the text.
- Line 22: When referring to 6-GIS, do you mean Gastrointestinal Severity Index? It might be helpful to define what you mean the first time it is used.
- Line 22: "bacterial markers: Bacteroidetes, Firmicutes, Actinomycetes" it would be clearer to write "bacterial markers, including Bacteroidetes, Firmicutes, and Actinomycetes".
- Check throughout the text that genus and species names are always written in italics.
- Line 28-29: “Compared to asymptomatic ones” a better alternative could be “compared to those who were asymptomatic” to make the sentence flow more smoothly.
- Line 30: “physiopathology” is less common than “pathophysiology”. Therefore, replace it with “pathophysiology” for greater consistency.
- Line 38: Rewrite the sentence “to acquiring other comorbidities throughout life”.
- Line 39: What do the percentages refer to? Are unclear.
- Line 41-46: To be revised to avoid repetition of gastrointestinal symptoms. Please rewrite it for clarity.
- Line 47: Remove the period.
- Line 50: Modify 'dietary deviation' to 'food selectivity'; it is a more precise.
- Line 53: What is meant by humidity?
- Line 55-56: Modify the term 'fecal bacterial genome' to 'metagenomics'; it is a more precise.
- Line 56-57: What does a poorly abundant-diversified mean? Rephrase the sentence to make it clearer.
- Line 58: It is recommended to consistently use the term autism or the acronym ASD throughout the text to assist the reader in comprehension.
- Line 71: Modify the term ‘early stages of human development’ with ‘since birth’.
- Merge paragraph 2.1 with the description of sociodemographic data and health indicators into a single paragraph, as it always describes the study population. The paragraph could be renamed as 'Patient Characteristics'.
- Rename paragraph 2.2 as 'Methods and Assessments'.
- Line 108: What does stool washing protocol mean?Does it refer to the protocol for the pre-treatment of the fecal sample?
- Line 122: Change the colon with a comma.
- Line 125: Remove the period between 16 and the S.
- Merge paragraph 2.2.4 with 2.2.5 as both describe PCRs used to amplify the 16S gene. It could be renamed as: 16S rRNA Gene Amplification (2S-PCR).
- Line 129: If the acronym NT has not been used earlier, spell out the full term.
- The readability could be improved by presenting the PCR protocol in bullet points. Also, I would not use arrows to indicate the increase or decrease of the PCR binding conditions.
- Line 141: The Tm temperature is missing.
- Line 148: Move Figure 1 to the results section, describing the results, as it is not the correct section to place the figure.
- Line 159: Explain the sentence better. What does 211 stands for?
- For all figures and tables, place the identifier (e.g., Table 1. Descriptive data of participants) on the same line as the description (e.g., lines 185-187), as this way it is clear that it refers to the description of the figure or table.
- Format and standardize the styles of all the tables.
- In Table 2, in the variable rows, insert FGID+ and FGID-, in order to make it immediately clear what is being referred to.
- Remove Table 3 from the text and place it in the supplementary materials.
- Advice to keep only the first table, which contains the statistically significant data, and move the others to the supplementary materials to maintain focus on the text and make the reading flow more smoothly.
- Line 212-215: If the data refer to Table 1, they should be moved to where the table data are described.
- Line 218: Remove the period.
- Paragraph 3.4 should be moved after the general description of the study group (paragraph 3.1) so that it concludes the general description of the participants before transitioning to the description of symptoms and pre-natal and perinatal information.
- Line 331: Remove the 's' from microbiota.
- Results and discussion should be separated into two paragraphs (Paragraph 3: Results and Paragraph 4: Discussion), as it is necessary to first describe the data and then make inferences for the discussion. Therefore, the conclusions will become Paragraph 5.
Must be improved. At multiple locations, the sentence is incomplete and in other cases they are unclear.
Author Response
March, 2025
Ms. Joyce Qiu
Section Managing Editor
Gastroenterology Insight
Thank you for coordinating and handling the manuscript review process. Additionally, to the suggested changes, we are proposing an adjustment to the title: “Colonic dysbiosis is associated with gastrointestinal disorders more than mother-child epidemiological traits in Mexican children with autism spectrum disorders”- gastroent-3482138. We really appreciate the opportunity to address the editor´s comments; so, we are submitting R1-gastroent-3482138 in response to your communication.
We provide below our responses to the editor’s/reviewer’s comments. We are submitting the reviewed manuscript (red marked), considering carefully all comments and incorporating all changes required to improve the manuscript quality and suitability for publication. Additionally, English was improved by a specialist (changes not red-marked); take into consideration that some of the changes suggested by the reviewers and that were attended on redaction were also modified during this edition.
Best regards,
Dr. Rocio Campos-Vega
Researcher-professor. Research and Graduate Studies in Food Science,
School of Chemistry.
Universidad Autónoma de Querétaro
C.U., Cerro de las Campanas,
Querétaro, Qro., México. CP 76010
+52 (442) 19212 00 Ext. 5590
e-mail: chio_cve@yahoo.com.mx
Reviewer 1
- Line 19: It might be clearer to use the term “gut microbiota (GM)” instead of colonic-fecal microbiota. To be revised throughout the rest of the manuscript.
Our response: Thanks for the observation- the changes was made along the text.
- Line 20-21: "A cases FGIDs (+) and controls FGIDs (-)” is not very clear; you could write “a study of FGIDs (+) cases and FGIDs (-) controls” for a clearer formulation.
Our response: Thanks- the redaction was modified (lines 20-21)
- Line 22: "Distal gastrointestinal pathologies” could simply say “Gastrointestinal disorders” to improve the flow of the text.
Our response: Thanks- the redaction was modified. (line 21)
- Line 22: When referring to 6-GIS, do you mean Gastrointestinal Severity Index? It might be helpful to define what you mean the first time it is used.
Our response: The definition of it was included. (line 22)
- Line 22: "bacterial markers: Bacteroidetes, Firmicutes, Actinomycetes" it would be clearer to write "bacterial markers, including Bacteroidetes, Firmicutes, and Actinomycetes".
Our response: Thanks- the redaction was modified. (lines 23 and 24)
- Check throughout the text that genus and species names are always written in italics.
Our response: The full document was reviewed.
- Line 28-29: “Compared to asymptomatic ones” a better alternative could be “compared to those who were asymptomatic” to make the sentence flow more smoothly.
Our response: Thanks- the redaction was modified. (lines 29-30)
- Line 30: “physiopathology” is less common than “pathophysiology”. Therefore, replace it with “pathophysiology” for greater consistency.
Our response: Thanks- the redaction was modified. (line 31)
- Line 38: Rewrite the sentence “to acquiring other comorbidities throughout life”.
Our response: Thanks- the redaction was modified. (lines 37 and 38)
- Line 39: What do the percentages refer to? Are unclear.
Our response: The information was expanded. (lines 38 and 39)
- Line 41-46: To be revised to avoid repetition of gastrointestinal symptoms. Please rewrite it for clarity.
Our response: Thanks- the redaction was modified. (lines 42-47)
- Line 47: Remove the period.
Our response: Thanks- the redaction was modified.
- Line 50: Modify 'dietary deviation' to 'food selectivity'; it is a more precise.
Our response: Thanks- the redaction was modified. (line 50)
- Line 53: What is meant by humidity?
Our response: Thanks- the redaction was modified.
- Line 55-56: Modify the term 'fecal bacterial genome' to 'metagenomics'; it is a more precise.
Our response: Thanks- the redaction was modified. (line 55)
- Line 56-57: What does a poorly abundant-diversified mean? Rephrase the sentence to make it clearer.
Our response: Thanks- the redaction was modified. (line 56)
- Line 58: It is recommended to consistently use the term autism or the acronym ASD throughout the text to assist the reader in comprehension.
Our response: Thanks- the text was modified.
- Line 71: Modify the term ‘early stages of human development’ with ‘since birth’.
Our response: Thanks- the text was modified.
- Merge paragraph 2.1 with the description of sociodemographic data and health indicators into a single paragraph, as it always describes the study population. The paragraph could be renamed as 'Patient Characteristics'.
Our response: Thanks- the text was modified. (line 75)
- Rename paragraph 2.2 as 'Methods and Assessments'.
Our response: Thanks for the suggestion- the text was modified. (line 89)
- Line 108: What does stool washing protocol mean? Does it refer to the protocol for the pre-treatment of the fecal sample?
Our response: The text was modified to avoid confusion. (line 104)
- Line 122: Change the colon with a comma.
Our response: Thanks- the text was modified.
- Line 125: Remove the period between 16 and the S.
Our response: The text was modified.
- Merge paragraph 2.2.4 with 2.2.5 as both describe PCRs used to amplify the 16S gene. It could be renamed as: 16S rRNA Gene Amplification (2S-PCR).
Our response: Thanks- the text was modified. (line 120)
- Line 129: If the acronym NT has not been used earlier, spell out the full term.
Our response: The text was modified.
- The readability could be improved by presenting the PCR protocol in bullet points. Also, I would not use arrows to indicate the increase or decrease of the PCR binding conditions.
Our response: Thanks- the text was modified. (lines 130-144)
- Line 141: The Tm temperature is missing.
Our response: Thanks- the information was included. (line 134)
- Line 148: Move Figure 1 to the results section, describing
the results, as it is not the correct section to place the figure.
Our response: Thanks- the figures and tables were removed from the main document because the journal editors placed these there.
- Line 159: Explain the sentence better. What does 211 stands for?
Our response: Thanks- the text was modified. It was a typo.
- For all figures and tables, place the identifier (e.g., Table 1. Descriptive data of participants) on the same line as the description (e.g., lines 185-187), as this way it is clear that it refers to the description of the figure or table.
Our response: Thanks- the text was modified.
- Format and standardize the styles of all the tables.
In Table 2, in the variable rows, insert FGID+ and FGID-, in order to make it immediately clear what is being referred to.
Remove Table 3 from the text and place it in the supplementary materials.
Advice to keep only the first table, which contains the statistically significant data, and move the others to the supplementary materials to maintain focus on the text and make the reading flow more smoothly.
Line 212-215: If the data refer to Table 1, they should be moved to where the table data are described.
Line 218: Remove the period.
Our response: The text and tables were modified. some of them are included as supplementary material, and only tables including the information from children are kept as part of the main document.
- Paragraph 3.4 should be moved after the general description of the study group (paragraph 3.1) so that it concludes the general description of the participants before transitioning to the description of symptoms and pre-natal and perinatal information.
Our response: Thanks- the text was modified. (lines 174-198)
- Line 331: Remove the 's' from microbiota.
Our response: Thanks- the text was modified.
- Results and discussion should be separated into two paragraphs (Paragraph 3: Results and Paragraph 4: Discussion), as it is necessary to first describe the data and then make inferences for the discussion. Therefore, the conclusions will become Paragraph 5.
Our response: Thanks for the suggestion. In this case, we prefer to maintain the sections as first proposed. Thanks in advance for your understanding.
Reviewer 2
- Line 120: Which is the correct identification of the solution, PCA or PCIA?
Our response: Thanks- the text was modified. (line 115)
Figure 1: The graphics are too small, and the font size of the axis labels is also too small, making them difficult to read.
Our response: The format was improved.
- Figure 1: Consider presenting the figures in a different order for better clarity. Currently, there are three panels with four, three, and five graphs each. A configuration of three by four or four by three might be more effective.
Our response: We appreciate the suggestion; the format was improved to have a better configuration considering the classification of the results.
- Figure 1: The significance markers are unclear. You have marked them with letters, but sometimes you use "a-a" and other times "a-b," which is confusing. Could you use asterisks to indicate significant changes and provide the corresponding p-values?
Our response: We appreciate the suggestion; the format was improved for a better understanding.
- Caption for Figure 1, Line 159: In the sentence “Each box plot extends from 211 the 25th-75th percentile, median value (inner horizontal line), and whiskers (dots) show the highest and lowest values (excluding outliers)” the number 211 appears to be a typo. Please clarify its meaning or correct it.
Our response: The text was corrected.
- Line 193: The sentence “The presence of obstetric comorbidities was high among the mothers: 42 women (95%) presented between 1 - 8 comorbidities (mean 3 ± 2.5)” is unclear. Higher compared to whom?
Our response: The text was corrected. (lines 201-202)
- Line 330 and throughout the manuscript: Refer to patients as “children with autism” or “patients with ASD.” Avoid using the term “autistics.”
Our response: Thanks- the text was modified.
- Line 335: Complete the sentence with “patients with autism”; the word “autism” is missing.
Our response: Thanks- the text was modified.
- Figure 2: The same comments for Figure 1 apply here.
Our response: Thanks- the text was modified.
Reviewer 2 Report
Comments and Suggestions for AuthorsComments on Version 1 of “Colonic Dysbiosis, More Than Mother-Child Epidemiological Traits, Is Associated with Gastrointestinal Disorders in Mexican Children with Autism Spectrum Disorders”
Line 120: Which is the correct identification of the solution, PCA or PCIA?
Figure 1: The graphics are too small, and the font size of the axis labels is also too small, making them difficult to read.
Figure 1: Consider presenting the figures in a different order for better clarity. Currently, there are three panels with four, three, and five graphs each. A configuration of three by four or four by three might be more effective.
Figure 1: The significance markers are unclear. You have marked them with letters, but sometimes you use "a-a" and other times "a-b," which is confusing. Could you use asterisks to indicate significant changes and provide the corresponding p-values?
Caption for Figure 1, Line 159: In the sentence “Each box plot extends from 211 the 25th-75th percentile, median value (inner horizontal line), and whiskers (dots) show the highest and lowest values (excluding outliers)” the number 211 appears to be a typo. Please clarify its meaning or correct it.
Line 193: The sentence “The presence of obstetric comorbidities was high among the mothers: 42 women (95%) presented between 1 - 8 comorbidities (mean 3 ± 2.5)” is unclear. Higher compared to whom?
Line 330 and throughout the manuscript: Refer to patients as “children with autism” or “patients with ASD.” Avoid using the term “autistics.”
Line 335: Complete the sentence with “patients with autism”; the word “autism” is missing.
Figure 2: The same comments for Figure 1 apply here.
Author Response
March, 2025
Ms. Joyce Qiu
Section Managing Editor
Gastroenterology Insight
Thank you for coordinating and handling the manuscript review process. Additionally, to the suggested changes, we are proposing an adjustment to the title: “Colonic dysbiosis is associated with gastrointestinal disorders more than mother-child epidemiological traits in Mexican children with autism spectrum disorders”- gastroent-3482138. We really appreciate the opportunity to address the editor´s comments; so, we are submitting R1-gastroent-3482138 in response to your communication.
We provide below our responses to the editor’s/reviewer’s comments. We are submitting the reviewed manuscript (red marked), considering carefully all comments and incorporating all changes required to improve the manuscript quality and suitability for publication. Additionally, English was improved by a specialist (changes not red-marked); take into consideration that some of the changes suggested by the reviewers and that were attended on redaction were also modified during this edition.
Best regards,
Dr. Rocio Campos-Vega
Researcher-professor. Research and Graduate Studies in Food Science,
School of Chemistry.
Universidad Autónoma de Querétaro
C.U., Cerro de las Campanas,
Querétaro, Qro., México. CP 76010
+52 (442) 19212 00 Ext. 5590
e-mail: chio_cve@yahoo.com.mx
Reviewer 1
- Line 19: It might be clearer to use the term “gut microbiota (GM)” instead of colonic-fecal microbiota. To be revised throughout the rest of the manuscript.
Our response: Thanks for the observation- the changes was made along the text.
- Line 20-21: "A cases FGIDs (+) and controls FGIDs (-)” is not very clear; you could write “a study of FGIDs (+) cases and FGIDs (-) controls” for a clearer formulation.
Our response: Thanks- the redaction was modified (lines 20-21)
- Line 22: "Distal gastrointestinal pathologies” could simply say “Gastrointestinal disorders” to improve the flow of the text.
Our response: Thanks- the redaction was modified. (line 21)
- Line 22: When referring to 6-GIS, do you mean Gastrointestinal Severity Index? It might be helpful to define what you mean the first time it is used.
Our response: The definition of it was included. (line 22)
- Line 22: "bacterial markers: Bacteroidetes, Firmicutes, Actinomycetes" it would be clearer to write "bacterial markers, including Bacteroidetes, Firmicutes, and Actinomycetes".
Our response: Thanks- the redaction was modified. (lines 23 and 24)
- Check throughout the text that genus and species names are always written in italics.
Our response: The full document was reviewed.
- Line 28-29: “Compared to asymptomatic ones” a better alternative could be “compared to those who were asymptomatic” to make the sentence flow more smoothly.
Our response: Thanks- the redaction was modified. (lines 29-30)
- Line 30: “physiopathology” is less common than “pathophysiology”. Therefore, replace it with “pathophysiology” for greater consistency.
Our response: Thanks- the redaction was modified. (line 31)
- Line 38: Rewrite the sentence “to acquiring other comorbidities throughout life”.
Our response: Thanks- the redaction was modified. (lines 37 and 38)
- Line 39: What do the percentages refer to? Are unclear.
Our response: The information was expanded. (lines 38 and 39)
- Line 41-46: To be revised to avoid repetition of gastrointestinal symptoms. Please rewrite it for clarity.
Our response: Thanks- the redaction was modified. (lines 42-47)
- Line 47: Remove the period.
Our response: Thanks- the redaction was modified.
- Line 50: Modify 'dietary deviation' to 'food selectivity'; it is a more precise.
Our response: Thanks- the redaction was modified. (line 50)
- Line 53: What is meant by humidity?
Our response: Thanks- the redaction was modified.
- Line 55-56: Modify the term 'fecal bacterial genome' to 'metagenomics'; it is a more precise.
Our response: Thanks- the redaction was modified. (line 55)
- Line 56-57: What does a poorly abundant-diversified mean? Rephrase the sentence to make it clearer.
Our response: Thanks- the redaction was modified. (line 56)
- Line 58: It is recommended to consistently use the term autism or the acronym ASD throughout the text to assist the reader in comprehension.
Our response: Thanks- the text was modified.
- Line 71: Modify the term ‘early stages of human development’ with ‘since birth’.
Our response: Thanks- the text was modified.
- Merge paragraph 2.1 with the description of sociodemographic data and health indicators into a single paragraph, as it always describes the study population. The paragraph could be renamed as 'Patient Characteristics'.
Our response: Thanks- the text was modified. (line 75)
- Rename paragraph 2.2 as 'Methods and Assessments'.
Our response: Thanks for the suggestion- the text was modified. (line 89)
- Line 108: What does stool washing protocol mean? Does it refer to the protocol for the pre-treatment of the fecal sample?
Our response: The text was modified to avoid confusion. (line 104)
- Line 122: Change the colon with a comma.
Our response: Thanks- the text was modified.
- Line 125: Remove the period between 16 and the S.
Our response: The text was modified.
- Merge paragraph 2.2.4 with 2.2.5 as both describe PCRs used to amplify the 16S gene. It could be renamed as: 16S rRNA Gene Amplification (2S-PCR).
Our response: Thanks- the text was modified. (line 120)
- Line 129: If the acronym NT has not been used earlier, spell out the full term.
Our response: The text was modified.
- The readability could be improved by presenting the PCR protocol in bullet points. Also, I would not use arrows to indicate the increase or decrease of the PCR binding conditions.
Our response: Thanks- the text was modified. (lines 130-144)
- Line 141: The Tm temperature is missing.
Our response: Thanks- the information was included. (line 134)
- Line 148: Move Figure 1 to the results section, describing
the results, as it is not the correct section to place the figure.
Our response: Thanks- the figures and tables were removed from the main document because the journal editors placed these there.
- Line 159: Explain the sentence better. What does 211 stands for?
Our response: Thanks- the text was modified. It was a typo.
- For all figures and tables, place the identifier (e.g., Table 1. Descriptive data of participants) on the same line as the description (e.g., lines 185-187), as this way it is clear that it refers to the description of the figure or table.
Our response: Thanks- the text was modified.
- Format and standardize the styles of all the tables.
In Table 2, in the variable rows, insert FGID+ and FGID-, in order to make it immediately clear what is being referred to.
Remove Table 3 from the text and place it in the supplementary materials.
Advice to keep only the first table, which contains the statistically significant data, and move the others to the supplementary materials to maintain focus on the text and make the reading flow more smoothly.
Line 212-215: If the data refer to Table 1, they should be moved to where the table data are described.
Line 218: Remove the period.
Our response: The text and tables were modified. some of them are included as supplementary material, and only tables including the information from children are kept as part of the main document.
- Paragraph 3.4 should be moved after the general description of the study group (paragraph 3.1) so that it concludes the general description of the participants before transitioning to the description of symptoms and pre-natal and perinatal information.
Our response: Thanks- the text was modified. (lines 174-198)
- Line 331: Remove the 's' from microbiota.
Our response: Thanks- the text was modified.
- Results and discussion should be separated into two paragraphs (Paragraph 3: Results and Paragraph 4: Discussion), as it is necessary to first describe the data and then make inferences for the discussion. Therefore, the conclusions will become Paragraph 5.
Our response: Thanks for the suggestion. In this case, we prefer to maintain the sections as first proposed. Thanks in advance for your understanding.
Reviewer 2
- Line 120: Which is the correct identification of the solution, PCA or PCIA?
Our response: Thanks- the text was modified. (line 115)
Figure 1: The graphics are too small, and the font size of the axis labels is also too small, making them difficult to read.
Our response: The format was improved.
- Figure 1: Consider presenting the figures in a different order for better clarity. Currently, there are three panels with four, three, and five graphs each. A configuration of three by four or four by three might be more effective.
Our response: We appreciate the suggestion; the format was improved to have a better configuration considering the classification of the results.
- Figure 1: The significance markers are unclear. You have marked them with letters, but sometimes you use "a-a" and other times "a-b," which is confusing. Could you use asterisks to indicate significant changes and provide the corresponding p-values?
Our response: We appreciate the suggestion; the format was improved for a better understanding.
- Caption for Figure 1, Line 159: In the sentence “Each box plot extends from 211 the 25th-75th percentile, median value (inner horizontal line), and whiskers (dots) show the highest and lowest values (excluding outliers)” the number 211 appears to be a typo. Please clarify its meaning or correct it.
Our response: The text was corrected.
- Line 193: The sentence “The presence of obstetric comorbidities was high among the mothers: 42 women (95%) presented between 1 - 8 comorbidities (mean 3 ± 2.5)” is unclear. Higher compared to whom?
Our response: The text was corrected. (lines 201-202)
- Line 330 and throughout the manuscript: Refer to patients as “children with autism” or “patients with ASD.” Avoid using the term “autistics.”
Our response: Thanks- the text was modified.
- Line 335: Complete the sentence with “patients with autism”; the word “autism” is missing.
Our response: Thanks- the text was modified.
- Figure 2: The same comments for Figure 1 apply here.
Our response: Thanks- the text was modified.
