Antimicrobial Resistance Distribution and Quorum-Sensing Regulation of Enterococcal Strains, Isolated from Hospitalized Patients

Round 1

Reviewer 1 Report

The manuscript studied "Antimicrobial resistance distribution and quorum-sensing regulation of enterococcal strains, isolated from hospitalized patients" Overall, the study is good, however this is a limited prevalence study, the authors only detect susceptibility profiles and quorum-sensing regulation genes in enterococcal strains. It would be better if they also do some mechanistic study, For example. MLST, Biofilm, and conjugation to check the dissemination of these quorum-sensing regulation genes among enterococcal strains.   Regarding this study, it is necessary to mention the following points, and if corrections are made, it can be considered for publication.
1. There are typographical errors throughout the text. 2. Why the authors only focusing on few resistance genes what was the criteria 3. How can we use the current results as predictive models for identifying emergence of such infections. 4. As the samples were obtained from two different hospitals so how many were "E. faecalis and E. faecium" from each hospital? Did you find some related isolates. 5. Why the authors not done MLST typing to further characterize the isolates relationship 6. Why the authors choose only 7 antibiotics for susceptibility testing. Why only DDST testing were used for susceptibility why not other assays like MIC.? 7. Discussion part should be in context with your results, explain only your important results.  8. Also write the gaps and limitations of your study in your discussion part. 9. The tables should be made in format according to the journal 10. In Table 5 and Table 7 I am not clear about the Chi Square results "0,266/ 3,841, asa1 " 314,544 / 12,592" etc.........Please explain these values.
11. Be sure to refer to recent studies in the discussion section.
12. The authors must check the style of the original articles and make them according to the journal template    

Author Response

Response to Reviewer 1 Comments

 

In order to clarify the information, we have provided, we have made revisions as you recommended. The revisions were highlighted by using the "Track Changes" function in Microsoft Word, so that changes are easily visible. Tables and article style were made according to the journal template.

    

Point 1. There are typographical errors throughout the text.

Response 1. The typographical errors throughout the text were corrected.

The number of enterococci by species from the two hospitals is noted.

 

Point 2. Why the authors only focusing on few resistance genes what was the criteria?

Response 2. We focus precisely on these antimicrobial resistance genes because they are responsible for resistance to the main groups of antibiotics used to treat enterococcal infections.

Point 3. How can we use the current results as predictive models for identifying emergence of such infections?

Response 3. The possible explanation is to use the data collected to create a neural network, but this is beyond the scope of our study, so we have removed this suggestion from the text.

Point 4. As the samples were obtained from two different hospitals so how many were "E. faecalis and E. faecium" from each hospital?

Response 4. The number of enterococci by species from the two hospitals was included in the text, (point 2.1, section Material and methods).

Point 5. Why the authors not done MLST typing to further characterize the isolates relationship?

Response 5. MLST was not performed because we choose to use methods readily available in regular microbiology laboratories.

Point 6. Why the authors choose only 7 antibiotics for susceptibility testing. Why only DDST testing were used for susceptibility why not other assays like MIC?

Response 6. We applied only these 7 antibiotics for susceptibility testing, because we work according to a EUCAST standard. According to the EUCAST standard MIC does not have to be used for antimicrobial susceptibility testing, but instead DDST can be used.

Point 7. Discussion part should be in context with your results, explain only your important results.

Response 7. We have made corrections to be more focussed on our results.

Point 8. Also write the gaps and limitations of your study in your discussion part.

Response 8. We have included some inherent limitations that PCR assays possess.

Point 9. The tables should be made in format according to the journal 

Response 9. We have made corrections.

Point 10. In Table 5 and Table 7 I am not clear about the Chi Square results "0,266/ 3,841, asa1 " 314,544 / 12,592" etc.........Please explain these values.

Response 10. The significantly higher observed chi square values than the critical chi square value prove the statistical significance of the experimental data. The greater the value of the observed chi function when compared to the critical one, the lower the value of P, the greater the statistical significance of the results.

Point 11 Be sure to refer to recent studies in the discussion section.

Response 11. We have compared our results with the most recent research possible.

Point 12 The authors must check the style of the original articles and make them according to the journal template  

Response 12. We have used the journal template to prepare our manuscript.

Reviewer 2 Report

Although Enterococcus is a normal flora of the gastrointestinal and urogenital systems, the bacteria can nevertheless lead to serious infections such as bacteraemia, endocarditis, urinary tract infections, and wounds. Currently, infections caused by antibiotic-resistant strains of Enerococcus faecalis and Enterococcus faecium are a serious problem for the healthcare system.

The aim of the study was to evaluate the pattern of antimicrobial resistance and to analyze the frequency of quorum-sensing asa1 and esp genes in clinical isolates representing the genus Enterococcus.

Remarks

Page 1 line 36 – Enterococcus – the Latin name should be written in italics

Materials and Methods 2.1. Sample collection, isolation, and identification - please provide the genes that were used for molecular identification.

I understand that the asa1 and esp genes were analyzed. Based on the data from the publication, wouldn't it be better to use the gelE gene in the identification, as it is more frequently present in Enterococcus spp. strains?

Pages 2 line 87- "5 The inhibition .." - please delete the number 5

"VanA", "VanB" – the genes are written in lower case and italics and with a capital letter if we write about vancomyco-resistance type A (VanA) and vancomyco-resistance type B

References item 61 – the font should be reduced

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 2 Comments

In order to clarify the information, we have provided in our manuscript, we have made revisions as you recommended. The revisions were highlighted by using the "Track Changes" function in Microsoft Word, so that changes are easily visible. We have changed the text in the Introduction and the Aim according your recommendations.

Point 1

Response 1. We wrote the Enterococcus in italics.

Point 2

Response 2. We provided the gene that we used for molecular identification.

Point 3

Response 3. In our previous study, published in Trakia journal of Sciences we have already looked at the gelE gene, so in the current study, we have decided to examine the other two genes asa1 and esp.

Point 4

Response 4. We deleted the number 5.

Point 5

Response 5. We corrected vanA and vanB where the where necessary.

Point 6

Response 6. We reduced the font of reference item 61.

Round 2

Reviewer 1 Report

The authors have successfully address all the comments and suggestions I have made. Therefore, I would recommend this for publication.