Environmental Dissemination of Antimicrobial Resistance: A Resistome-Based Comparison of Hospital and Community Wastewater Sources

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript entitles “Environmental dissemination of antimicrobial resistance: A 2 resistome-based comparison of hospital and community 3 wastewater sources” by Kitano et al.. This manuscript focused on the study of ARG surveillance in wastewater, comparison of wastewater sources from hospital and shopping mall. Short-read sequencing was applied to study the amount of ARGs in wastewater samples and compare RPKM values to analyses the abundance of ARGs. The results showed that ARGs were found in hospital wastewater more than shopping mall, especially, aminoglycoside resistance genes and beta-lactam resistance genes. No ARGs from shopping mall was higher than hospital with significant difference. The wastewater samples were determined the residual of antimicrobial concentrations using UPLC-MS/MS and found that levofloxacin and clarithromycin were detected in hospital and shopping mall. Levofloxacin showed higher concentration than clarithromycin. The content is very interested in wastewater surveillance field, in both ARGs and antimicrobial concentrations in wastewater. The quality of presentation is excellent. It has some minor points that require to improve manuscript. And my comments are below; I hope that these comments might help authors to improve manuscript and get ready to be published.

Major comment:

• Please give the results of UPLC-MS/MS and emphasized the levofloxacin and clarithromycin from wastewater sample from hospital and shopping mall.

Minor comments:

• In line 126 – 133, that information is not significant difference and it already showed in supplementary table S1. Authors is not need to explain in main text, cited the table S1 is enough.
• In line 138, please give a citation for a previous study.
• In line 146, it should be December 2021 (1.9 log10) and February 2021 (1.7 log10) were notable peaks?
• In supplementary tables, please repeat the header row in every page.
• In figure S2, please re-check label on x-axis.
• In figure 2b, please re-check label on y-axis. If data in figure 2b is linked to table S4, It seen like the unit should be ratio, not log10?
• In table 1, please re-type the name of genes as in main text.

Author Response

Please see the attachment.

Author Response File: Author Response.doc

Reviewer 2 Report

Comments and Suggestions for Authors

Overall, it is an interesting and straightforward study! A few comments need to be addressed:

1. Figure 1 can be improved, it is currently blurred and hard to read due to the font size and vague words; please also add legends in Figure 1.
2. Figure 2b, did the authors conduct any statistical analyses between Clarithromycin and Levofloxacin?
3. Line 284, why samples were collected three times during this time? What factors justify the sample collection dates?
4. how "small" the community is? Is the hospital located within the community sewershed?

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

1. Fig 1, 2 and Table 1 didn’t appear right after mentioning
2. Fig 1, there is the figure title in the plot, and on the bottom
3. Line 137-138, a reference citation of the previous study?
4. Keep it consistent in the results, use one decimal or two? E.g. line 144, -0.53 to 1.9
5. In the section 4.1, prove the distance between the hospital and the mall? If possible provide a map for the sampling location?

Author Response

Please see the attachment.

Author Response File: Author Response.docx