PONE-D-22-00832Association between microbial composition, diversity, and function of the maternal gastrointestinal microbiome with impaired glucose tolerance on the Glucose Challenge TestPLOS ONE

Dear Dr. Dreisbach,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

==============================Thank you for submitting this manuscript. Firstly, I must apologise for the time it has taken to deal with the manuscript - this was caused by initial difficulties in recruiting sufficiently qualified reviewers. The manuscript has divided opinion among the reviewers. Reviewer #3 believes that the manuscript is fine as is. Reviewer #1 has some relatively minor points which need to be dealt with in any revised manuscript. However, reviewer #2 has some major concerns with the manuscript which need to be dealt with robustly for it to be accepted for publication. Of these I believe that the key points that need revision are the ones concerning the inclusion and exclusion of participants, pregnancy outcomes, and consideration of potential confounders. I also have one additional very minor point which I believe merits revision: in lines 47, 181, 189, 207 & Table 3 please change each instance of "(p=<0.00)" to "(p<0.01)" or use exact p-values.

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Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

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Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

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Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

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Reviewer #2: No

Reviewer #3: Yes

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Reviewer #1: This is a well written and interesting cross-sectional study in an understudied population of women (pregnant women with GDM). This reviewer only has minor suggestions to the authors.

Line 35: Define “IGT” as an abbreviation here (i.e., “…impaired glucose tolerance (IGT)…”

Line 41: Define “NGT” as an abbreviation (e.g., “…and those without (normal glucose tolerance – NGT)…”)

Line 104-105: Please give a brief description of the heating/vortex protocols used to extract DNA from the swabs (e.g., temperatures used, length of incubation, etc.)

Lines 142-154: Was false discovery rate correction used? I would recommend FDR correction of your p-values because of the large number of comparisons done for this study; the FDR corrected results can be reported after the un-corrected results.

Lines 191-193: It is very difficult to see the difference in abundance of these bacteria between IGT and NGT because of how the data is presented. I would suggest using boxplots or log-fold-change bar plots to more clearly show the differences in abundance between IGT and NGT. Please also mention the results for antibiotic use and BMI category in-text and display those results in a boxplot/bar plot or remove them from the figure if they won’t be discussed.

Line 211-214: There are two “Table 2”s. The results in the first Table 2 (alpha diversity vs. BMI category) should be discussed in the text. The second Table 2 (glucose challenge vs alpha diversity) should be Table 3. The second table 2 is also missing p values and lacks in-text discussion of some of the results.

Lines 202-204: Results unclear. As stated, it sounds like Shannon/Inverse Simpson was negatively correlated with glucose challenge test, but the table shows a weakly positive correlation coefficient for both comparisons (0.09 and 0.10, respectively).

Lined 209-120: Remove the last sentence. Put “(Figure 2)” at the end of the previous sentence (i.e., …values in the impaired tolerance group (Figure 2).”). In the figure, please add a label to the x axis (e.g., “Does participant have IGT?”).

Reviewer #2: There are many studies about the intestinal flora of gestational diabetes mellitus. There are few studies on gestational impaired glucose tolerance. However, due to the mild severity of abnormal glucose tolerance during pregnancy, clinical research is of little significance

There are more statistical methods of intestinal flora that can be used

Lack of inclusion criteria and exclusion criteria for patients. No complete clinical data and pregnancy outcomes were provided, and no consideration was given to the situation that may affect the composition of intestinal flora, such as eating habits, application of antibiotics and exclusion of complications and complications during pregnancy

The discussion part directly cites the content of the result part. The reasoning and demonstration process part is lack of organization and supporting literature, which needs to be further summarized.

Reviewer #3: An interesting topic. Microbiota is very important for health. Glucose intolerance or impaired glucose metabolism alter the mintestinal microbiome, which in turn has consequences for the health of mother and child

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Reviewer #2: No

Reviewer #3: No

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Association between microbial composition, diversity, and function of the maternal gastrointestinal microbiome with impaired glucose tolerance on the Glucose Tolerance Test

PONE-D-22-00832R1

Dear Dr. Dreisbach,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Clive J. Petry, PhD

Academic Editor

PLOS ONE

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