Molecular characterization and transmission pattern of tetracycline resistance gene in tigecycline and carbapenem resistance klebsiella pneumoniae isolates at a tertiary care hospital India

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1. 

   Version published to 10.1099/acmi.0.001017.v2 on Access Microbiology

   Jul 7, 2025
2. 

   Access Microbiology

   May 22, 2025

   Dear authors, thank you for submitting your manuscript to Access Microbiology. It has now been reviewed by two experts in the field, whose comments are attached at the bottom of this email. Please review the language used in the manuscript - there are several aspects which would require revision to clarify the author's intended message. We offer a discounted translation service, if this could be of interest please find more information here (https://www.editage.com/; https://www.microbiologyresearch.org/submission-and-peer-review#6). It was also noted that accession numbers should be incldued. I welcome the authors to address these concerns as well as all other reviewer comments that will strengthen this manuscript that I believe will be of interest to the wider scientific community.

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3. 

   Access Microbiology

   May 16, 2025

   Comments to Author

   In this study Jyoti et al have looked into the molecular characterization and transmission pattern of tetracycline resistance gene in tigecycline and carbapenem resistance klebsiella pneumoniae isolates from a tertiary care hospital in India. The study is interesting; however the authors really need to put efforts to improve their work in a presentable format. The manuscript is not formatted and tenses are all mixed up. It looks like some of the sections have been copied and pasted from a dissertation/thesis. I also have following observations with the present version of manuscript The authors need to thoroughly revise the introduction section of the manuscript. The Objectives need to have connectivity with the main aim/hypothesis laid out in the introduction section. Demographic data- provide …

   Comments to Author

   In this study Jyoti et al have looked into the molecular characterization and transmission pattern of tetracycline resistance gene in tigecycline and carbapenem resistance klebsiella pneumoniae isolates from a tertiary care hospital in India. The study is interesting; however the authors really need to put efforts to improve their work in a presentable format. The manuscript is not formatted and tenses are all mixed up. It looks like some of the sections have been copied and pasted from a dissertation/thesis. I also have following observations with the present version of manuscript The authors need to thoroughly revise the introduction section of the manuscript. The Objectives need to have connectivity with the main aim/hypothesis laid out in the introduction section. Demographic data- provide information on the age range of the patients in the results section and not in the material and methods section. What was the incubation temperature used for antibiotic sensitivity tests? "Sequenced data was matched from NCBI data base and submitted in GenBank"- The database is INSDC and not NCBI. Please fix the errors. Also, I could not find the accession numbers. What was the amplicon length of tet(B)? Accession numbers are missing throughput the manuscript. For MALDI-TOF MS, critical information is missing. For example, peak positions (m/z) for each strain has to be included. What extraction procedure was adopted before the run was undertaken? Were the cultures grown overnight? The figures need re-working. The current figures need to be changed. I would strongly suggest the authors to do an alignment of blaNDM and blaOXA-48-like gene sequences which they reported in their study along with sequences reported from other published studies. This would also provide a clearer picture of the molecular evolution of these genes linked to resistance since critical information on plasmid backbone is missing.

   Please rate the manuscript for methodological rigour

   Poor

   Please rate the quality of the presentation and structure of the manuscript

   Poor

   To what extent are the conclusions supported by the data?

   Partially support

   Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

   No

   Is there a potential financial or other conflict of interest between yourself and the author(s)?

   No

   If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?

   Yes

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4. 

   Access Microbiology

   May 14, 2025

   Comments to Author

   This is an interesting study with matched sanger sequence and phenotype data and a good sample size (n = 152). It provides a useful dataset from a tertiary care hospital in Lucknow India showing the scale of carbapenem and tigecycline resistance. In general the English needs to be improved to aid readability. I think a large part needs to be rewritten in more readable language before accepting for publication. Some species names lack italics (mostly K. pneumoniae). However the science is robust, the authors are aware of the limitations of the study (that whole genome sequencing would have added to the study), and the findings are of interest to the wider scientific community. Line 2. klebsiella pneumoniae in title should be capitalised Line 45 - "tetracycline (tetA, tetB, tetK, tetM and tetS) …

   Comments to Author

   This is an interesting study with matched sanger sequence and phenotype data and a good sample size (n = 152). It provides a useful dataset from a tertiary care hospital in Lucknow India showing the scale of carbapenem and tigecycline resistance. In general the English needs to be improved to aid readability. I think a large part needs to be rewritten in more readable language before accepting for publication. Some species names lack italics (mostly K. pneumoniae). However the science is robust, the authors are aware of the limitations of the study (that whole genome sequencing would have added to the study), and the findings are of interest to the wider scientific community. Line 2. klebsiella pneumoniae in title should be capitalised Line 45 - "tetracycline (tetA, tetB, tetK, tetM and tetS) genes were performed by PCR" reword this - PCR was performed rather than the genes Line 46 - conjugation assay - change to assays Line 52 and others - "only 51.6% (16/31) and 29.1% (9/31) isolates" should be 29.1% (9/31) of isolates Line 56 - plasmids rather than plasmid? Line 60 - you could add to the conclusion that you have found the blaNDM and blaOXA to be transferable Line 65- "becoming common day by day in world and leading severe public health concern" - rewrite Line 70- "Despite of nosocomial infections" amend English Line 73 - Italicize K. pneumoniae The end of the introduction from line 99 onwards contains information on methods and some results (such as percentage of tetracycline resistance) which should be moved to the methods and results sections. The objectives of the study could be condensed from the bullet points or the whole section removed as the abstract covers these objectives. Line 140 - The split between male and female is interesting - it would be good to associate this metadata to each sample (KP1, KP2, etc.) and include more metadata - this can be included as a supplementary table Line 177 - clarify where these 152 carbapenem resistant Klebsiella pneumoniae isolates came from Line 180 - Figure 2 could be improved as it doesn't add anymore information than the text - a Sankey Diagram could be created where the number of CRKP flows to those that are tigecycline resistant and where each of these have come from Figure 3 - expand the figure capture to make it clear that these are percentages of tigecycline and carbapenem resistant K. pneumoniae isolates. It would also be good to include other resistance genes of interest in this figure such as those conferring resistance to aminoglycosides, phenicols, colistin, and other beta lactams. Since you have tested aztreonam, gentamicin, ciprofloxacin and colistin it would be good to see what resistance genes they have relating to this phenotypic resistance. Line 204 - just call it plasmids rather than "Extra-chromosomal genetic material"- there can be extra-chromosomal genetic material that are not plasmids. And I think you mean to say plasmid extractions were used as the template for PCR but this isn't clear in the text Line 215 "World wide" rather than "world wild" Line 216 acinetobacter baumannii is a species so should be italicised and the A capitalised Line 242 - "found that all patient infected with tet gene containing K. pneumoniae isolates" - reword this as you can't be infected by a gene but these patients were infected by the K. pneumoniae isolates containing the tet genes Line 244 - why were conjugation experiments only done for two of the four isolates? Line 249 - what do the numbers refer to in brackets after the genes - blaNDM (2/2) and blaOXA48 (2/3) - are these the number of transconjugants you obtained or the number of conjugation experiments that were run? It isn't clear in the text Line 262 - clarify precisely which online repositories these datasets can be found with associated URL

   Please rate the manuscript for methodological rigour

   Good

   Please rate the quality of the presentation and structure of the manuscript

   Poor

   To what extent are the conclusions supported by the data?

   Strongly support

   Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?

   No

   Is there a potential financial or other conflict of interest between yourself and the author(s)?

   No

   If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?

   Yes

   Read the original source
5. 

   Version published to 10.1099/acmi.0.001017.v1 on Access Microbiology

   Mar 5, 2025