Bacillus- and Lactobacillus-Based Dietary Synbiotics Are Associated with Shifts in the Oropharyngeal, Proximal Colonic, and Vaginal Microbiomes of Korean Native Black Pigs
Round 1
Reviewer 1 Report
- Please put the graph axes in black color.
- In "p-value" the p must be in italic.
- Line 108: Please add space after number (-20 °C).
- Line 150: Please provide R Studio program version.
Author Response
Point 1: Please put the graph axes in black color.
Response 1: First, we would like to thank the reviewer for taking the time to read and offer some helpful suggestions. As advised, the font color of graph axis texts has been changed to black in Figure 1, Figure2, Figure 3 and Figure 4. We believe that it is an improvement on the legibility of the axes.
Point 2: In "p-value" the p must be in italic.
Response 2: Thank you for pointing this out. We have italicized the ‘p’ in ‘p-value’ as recommended.
Point 3: Line 108: Please add space after number (-20 °C).
Response 3: We thank you for catching this typo. We have added the space between the number and the symbol in “-20 °C”.
Point 4: Line 150: Please provide R Studio program version.
Response 4: We thank the reviewer for highlighting this omission. Indeed, the R and R studio versions should have been declared in the methods. Accordingly, we have provided these versions in lines 150-151.
“Output from QIIME2 was imported into R (version 4.1.3 2022-03-10), in RStudio (2022.07.0+353) for further analysis”
Reviewer 2 Report
Interesting paper on the modulating effect of symbiotics (probiotics + prebiotics) on the oropharyngeal, proximal colonic and vaginal microbiomes of native Korean black pigs.
Appropriate methodology for the purpose of the study and appropriate statistical analysis for the data obtained.
Discussion and conclusions based on the results obtained.
Author Response
Point 1: Interesting paper on the modulating effect of symbiotics (probiotics + prebiotics) on the oropharyngeal, proximal colonic and vaginal microbiomes of native Korean black pigs.
Point 2: Appropriate methodology for the purpose of the study and appropriate statistical analysis for the data obtained.
Point 3: Discussion and conclusions based on the results obtained.
Response 3: We greatly appreciate the reviewer for carefully reading the manuscript and for their kind and positive comments.
Reviewer 3 Report
Dear Authors,
Thank you for allowing me to review your manuscript.
The introduction is well documented and the purpose of the study is clearly expressed.
At result section some correlations are always a good point.
However, the clinical implications should be better focused.
What could the new therapeutic strategies consist of?
Inaddition I would reserve a paragraph dedicated to possible clinical developments in humans and another to the limits of the study that are not adequately described.
Discussion and conclusions are well written.
Finally, for a multidisciplinary approach we propose to mention:
DOI10.1136/ijgc-2021-003241
DOI10.3390/biomedicines8120554
DOI10.3390/cancers14143457
Author Response
Dear Authors,
Thank you for allowing me to review your manuscript.
Point 1: The introduction is well documented, and the purpose of the study is clearly expressed. At result section some correlations are always a good point.
Response 1: We would like to thank the reviewer for taking the time to read through the manuscript and for providing some constructive suggestions and comments.
Point 2: However, the clinical implications should be better focused. What could the new therapeutic strategies consist of? In addition, I would reserve a paragraph dedicated to possible clinical developments in humans and another to the limits of the study that are not adequately described.
Response 2: We thank the reviewer for this constructive suggestion. As advised, we have edited the “Discussion” section to include a commentary on the “clinical implications “of this study in lines (450 - 468)
“Probiotics have had a growing application in maintenance of sexual and reproductive health in women (Lehtoranta2022). Although their application in clinical management of gynecological and obstetric conditions is a plausible prospect and a preferable alternative to antimicrobials, its efficacy remains inconclusive so far (Buggio2019). As such, further research remains necessary and use of the pig as a biomedical animal model facilitates such extensive studies given that pigs have physiological and anatomical similarities to humans (Pang2007). Our study explored an oral route of probiotic application which assumes that some of the ingested probiotic organisms can transit through the gastrointestinal tract and ascend into the vaginal tract from the rectum. Although this is more plausible in the pig than it is in humans for behavior reasons, changes in vaginal microbiome have been noted following oral probiotic regimens in women (Chen2021). Another mechanism through which oral probiotics could influence vaginal microbiota is through indirect systemic effects due to the probiotic’s modulatory effects within the gut microbiome (Cervantes-Barragan2017, Hou2022). Although it has been intuitively suggested that a topical, intra-vaginal application route may yield clearer results, both routes of administration have been found to be effective (Li2019). Overall, orally administered probiotics warrant attention since they are convenient to use and likely, more agreeable. Such a non-invasive application route, provides opportunities for the development of vaginal-microbiome-restoring probiotics following gynecological procedures especially when fertility sparing is a goal (Lehtoranta2022, Bergamini2022)”
About the limitations of the study, we added the following text under the “Conclusion” section in lines 479 - 485).
“However, this study had a few limitations including the following. First, the inclusion criteria for the experimental animals greatly limited the group sizes used in this study which slightly underpowered our findings. Second, our work was based on a metataxonomic approach and the shifts in predicted microbial functionality described herein only represent changes in potential functionality. To validate our findings, future studies with larger sample sizes should combine the metataxonomic strategy used here with meta-transcriptomic and metaproteomic approaches that can identify differential expression of microbial pathways and proteins.”
Point 3: Discussion and conclusions are well written.
Response 3: We thank the reviewer for this positive feed-back.
Point 4: Finally, for a multidisciplinary approach we propose to mention:
DOI10.1136/ijgc-2021-003241
DOI10.3390/biomedicines8120554
DOI10.3390/cancers14143457
Response 4: Thank you for these useful suggestions. Accordingly, we read and reviewed these studies into fertility-sparing ovarian surgeries. The studies were quite interesting and provided us with an insight into advances in clinical and surgical gynecology. In considerations of the hormonal influence of ovaries on the vaginal microbiomes, we do think that probiotic supplementation could be developed as a supportive therapy to facilitate recovery and restoration of vaginal microbial health after such fertility sparing gynecological interventions. This addition can be found in lines 466-468.
“Such a non-invasive application route, provides opportunities for the development of vaginal-microbiome-restoring probiotics following gynecological procedures especially when fertility sparing is a goal (Lehtoranta2022, Bergamini2022)”
Round 2
Reviewer 3 Report
The work has improved a lot.
Congratulations
