Azithromycin is a bacterial protein synthesis inhibitor of the drug class, Erythromycin and also used for treating intestinal infections. Since, KPN is also an intestinal bacteria we used this disc for susceptibility testing. Moreover, there are previous studies which also mentioned the use of Azithromycin against KPN Baker et. However, it has been removed from the list and figures of modified manuscript.
The list of antibiotics used for the test does not include any beta-lactamase inhibitor. We did not mention these in description, because the percentage of resistance or sensitivity was not significant compared to other antibiotics. It has been included in raw data.
However, it has also been added to the revised manuscript. A resistance analysis should be done according to the origin of the patient. Inpatients or outpatients. Response: Resistance for inpatient and outpatient has been analyzed in the revised draft. A figure containing the data has been added in the modified draft Figure 2, page Response: We selected the resistant isolates based on their antimicrobial resistant profile. Isolates showing resistance to the highest number of antibiotics were chosen for further analysis.
Isolates that had antibiotic resistant genes were further sent for gene sequencing. It has been added in the modified manuscript line- , page When speaking of a reservoir, it refers to a normal growth site.
It is suggested to modify the word by site of infection. Line to How did they show that they are treating the same Klebsiella clone as the patients infections? Response: In this study, Plasmid profiling showed a similarity between fomites and clinical isolates. Moreover, gene sequencing analyses showed that KPC-2 producing KPN strains were transferred from water to hospital and vice versa through mechanism.
This study investigated the clonal relationship of the strains through phylogram and MLST. A number of studies also previously reported similar findings Clarivet B, et al.
However, it has not been mentioned as a conclusive statement in the revised manuscript. This was stated as a probable explanation Line, Page- 17 -. In this study, we considered fomites for analyzing KPN isolates as well. We observed the presence of KPN isolates in sewerage water and bed trails from samples obtained from the neonatal ward.
This indicates that not only clinical specimens but fomites also act as sites of infection of KPN [48, 49]. Presence of KPN in non-biological samples might be a probable explanation of the phenomenon of horizontal transfer of transposable elements into KPN isolates and rapid transmission of a plasmid to another Enterobacteriaceae.
The study was carried out in two healthcare centers in Chattogram. The results could not be generalized to the southern Bangladesh region. Response: Authors would like to thank the reviewer for this important suggestion. Title has been modified. Figure 1A. Azithromycin should not be listed. AmoxiClav is not described in materials. Response: Azithromycin has been used commonly in KPN infections and antibiotic sensitivity tests in Bangladesh Jasmin et al.
In our study, it was according to the common practice of the hospitals of Bangladesh and patients' needs. In addition to that, it was used in KPN infections in some trials according to literature studies Getanda P et al. However, Azithromycin has been removed from the figure and the list in the modified draft.
Amoxiclav has been mentioned in the methodology in the revised manuscript page-8, Line Baker, K. Repurposing azithromycin and rifampicin against Gram-negative pathogens by combination with peptide potentiators. International journal of antimicrobial agents, 53 6 , Persisting transmission of carbapenemase-producing Klebsiella pneumoniae due to an environmental reservoir in a university hospital, France, to Differential expression of bla SHV related to susceptibility to ampicillin in Klebsiella pneumoniae.
Int J Antimicrob Agents. Epub Feb 2. PMID: Getanda, P. Short-term increase in the carriage of azithromycin-resistant Escherichia coli and Klebsiella pneumoniae in mothers and their newborns following intra-partum azithromycin: a post hoc analysis of a double-blind randomized trial.
JAC-Antimicrobial Resistance, 3 1 , dlaa Epub Mar Kim, J. Assessment of antibiotic resistance in Klebsiella pneumoniae exposed to sequential in vitro antibiotic treatments. Annals of clinical microbiology and antimicrobials, 15 1 , Jesmin Akter, A. American Journal of Drug Discovery and Development, 4: Science of The Total Environment.
Prevalence and molecular characterization of antibiotic resistance and associated genes in Klebsiella pneumoniae isolates: A clinical observational study in different hospitals in Chattogram, Bangladesh. Please, address the comments provider by the reviewer and resubmit as early as your convenience. Please submit your revised manuscript by Sep 18 PM. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references.
Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. Reviewer 2: The article sent for review has been improved in relation to the first version, however, some minor recommendations are made to improve it. Submitted filename: Reviewer recommendations 2. We thank the honorable reviewer for valuable feedback.
Please find our response below-. We have rechecked the manuscript and corrected the error. Lines to They are proper names of the units and the first letter of each word should. We have rechecked the manuscript and edited accordingly. When describing the primers used, it is not necessary to place F or R before the sequence, taking into account that they already indicate whether it is Forward or Reverse. It is recommended to place the name of the amplified gene in the case of the third pair of primers.
If the 5'-3' orientation is specified at the beginning of all primers, the extremities of each sequence could be eliminated. All this in order to improve the presentation and facilitate reading. Changes have been made accordingly. Figure 4. It would be interesting for the authors to mention the clonality of these analyzes. Try to hypothesize against this.
The analysis in sewerage water and bed trails from samples obtained from the neonatal Ward is mentioned. Did you make comparative analyzes between the KPN of samples with those of fomites? Was cross contamination found?.
It is recommended to discuss and conclude regarding the clonality found, this data is of interest. Response: We agree with the reviewer that clonality should be monitored. However, the fomites were assessed only to check the presence of KPN. But this needs more samples from fomites. And an in depth molecular study on fomite and source of contamination is needed.
We are working on the molecular identification of fomite and reservoir of the pathogen in the next phase of this study and that is a separate project. Considering the need of in depth analysis, we have excluded this part from the result and discussion of the modified manuscript. We believe this will help the readers to keep focus on the prevalence of antibiotic resistance in Klebsiella isolates from the clinic samples.
Response: Information regarding fomite has been excluded from the modified manuscript and the reason for this is explained in the above response. Submitted filename: Response to the reviewer. An invoice for payment will follow shortly after the formal acceptance.
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PLoS One. Published online Sep Mahbub Hasan. Monica Cartelle Gestal, Editor. Author information Article notes Copyright and License information Disclaimer. Competing Interests: The authors have declared that no competing interests exist. Received Mar 18; Accepted Aug This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
S1 Raw images: PDF. Attachment: Submitted filename: Reviewer recommendations. Attachment: Submitted filename: Reviewer recommendations 2.
Attachment: Submitted filename: Response to the reviewer. Attachment: Submitted filename: Reviewer recommendations 3. Abstract Objective This study was performed to investigate the prevalence of multidrug resistance and molecular characterization of Klebsiella pneumoniae KPN from clinical isolates in the southern region of Bangladesh.
Method The study was carried out using clinical isolates collected from two different hospitals of Chattogram. Table 1 Clinical characteristics of study population. Open in a separate window. Fig 1. Patterns of antimicrobial resistance among Klebsiella pneumonia isolates collected from two hospitals. Fig 2. Antibiotic sensitivity and resistance among different types of isolates.
Fig 3. Fig 5. Fig 4. Fig 6. Detection of plasmid by agarose gel electrophoresis. Acknowledgments The authors would like to thank the research assistants of Disease Biology and Molecular Epidemiology Research Group, Chattogram for their support during the study. Data Availability All relevant data are within the manuscript and its Supporting Information files.
References 1. Tackling antimicrobial resistance in Bangladesh: A scoping review of policy and practice in human, animal and environment sectors. PloS one. Antibiotic resistance of Escherichia coli isolated from poultry and poultry environment of Bangladesh. Internet Journal of food safety. Social-economic factors and irrational antibiotic use as reasons for antibiotic resistance of bacteria causing common childhood infections in primary healthcare.
Eur J Pediatr. Antibiotic resistance: global response needed. The Lancet Infectious Diseases. Antibiotic resistance in Bangladesh: A systematic review. International Journal of Infectious Diseases. Molecular characterization of multidrug-resistant Klebsiella pneumoniae isolates. Brazilian Journal of Microbiology. Situation of antibiotic resistance in Bangladesh and its association with resistance genes for horizontal transfer.
Genomic definition of hypervirulent and multidrug-resistant Klebsiella pneumoniae clonal groups. Emerging infectious diseases. Emerging invasive liver abscess caused by K1 serotype Klebsiella pneumoniae in Korea. Journal of Infection. Podschun R, Ullmann U. Klebsiella spp. Clinical microbiology reviews. Hypervirulent hypermucoviscous Klebsiella pneumoniae: a new and dangerous breed. Klebsiella pneumoniae: characteristics of carbapenem resistance and virulence factors.
Acta Biochimica Polonica. Determination of the frequency of carbapenemase producing Klebsiella pneumoniae isolates in Dhaka city , Bangladesh. Stamford Journal of Microbiology. Molecular analysis of antibiotic resistance determinants and plasmids in Malaysian isolates of multidrug resistant Klebsiella pneumoniae. Molecular characterization of clinical multidrug-resistant Klebsiella pneumoniae isolates. Annals of clinical microbiology and antimicrobials.
Global spread of carbapenemase-producing Enterobacteriaceae. Journal of antimicrobial chemotherapy. Infection, Genetics and Evolution. African Journal of Biotechnology. Journal of medical microbiology. Antimicrobial agents and chemotherapy. Applied and environmental microbiology. Journal of Infection and Chemotherapy.
Diversity and evolution of the class A chromosomal beta-lactamase gene in Klebsiella pneumoniae. Duplication of the chromosomal blaSHV gene in a clinical hypermutable strain of Klebsiella pneumoniae. Journal of Antimicrobial Chemotherapy.
Development and evaluation of a real-time PCR assay for detection of Klebsiella pneumoniae carbapenemase genes. Journal of clinical microbiology. Prevalence and molecular epidemiology of clinical isolates of Escherichia coli and Klebsiella pneumoniae harboring extended-spectrum beta-lactamase and carbapenemase genes in Bangladesh.
Microbial Drug Resistance. Virulent clones of Klebsiella pneumoniae: identification and evolutionary scenario based on genomic and phenotypic characterization. A gene, uge, is essential for Klebsiella pneumoniae virulence.
Infection and immunity. Journal of Bangladesh Academy of Sciences. A review on commonly used biochemical test for bacteria. Innovare Journal of Life Science. Clinical practice guideline for the management of candidiasis: update by the Infectious Diseases Society of America. Clinical Infectious Diseases.
Ahmed OB, Dablool A. Quality improvement of the DNA extracted by boiling method in gram negative bacteria. International Journal of Bioassays. Heat treatment of bacteria: a simple method of DNA extraction for molecular techniques. Kuwait Med J. MEGA7: molecular evolutionary genetics analysis version 7.
Molecular biology and evolution. Factors associated with Klebsiella bacteremia and its outcome in under-five children admitted with diarrhea. International journal of pediatrics. Isolation and identification of pathogenic Escherichia coli, Klebsiella spp. Status of ESBL producing bacteria isolated from skin wound at a tertiary care hospital in Bangladesh. Advances in Infectious Diseases. Multiple-antibiotic resistance mediated by plasmids and integrons in uropathogenic Escherichia coli and Klebsiella pneumoniae.
Bangladesh Journal of Microbiology. Prevalence and comparison of Beta-lactamase producing Escherichia coli and Klebsiella spp from clinical and environmental sources in Lahore, Pakistan.
African Journal of Microbiology Research. Klebsiella pneumonia with pneumatocele formation in a newborn infant. Canadian Medical Association Journal. Carbapenem resistant Enterobacteriaceae neonatal gut colonization: A future concern in healthcare settings.
Indian Journal of Microbiology Research. A structural view of the antibiotic degradation enzyme NDM-1 from a superbug. J Clin Diagn Res. Clinical Microbiology and Infection. Antimicrobial resistance in developing countries : Springer; Monica Cartelle Gestal , Academic Editor. Copyright and License information Disclaimer.
Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer s.
We look forward to receiving your revised manuscript. Please do not edit. Is the manuscript technically sound, and do the data support the conclusions? Has the statistical analysis been performed appropriately and rigorously? Review Comments to the Author Please use the space provided to explain your answers to the questions above. Please upload your review as an attachment if it exceeds 20, characters Reviewer 1: The manuscript by Tanni et al.
There is a duplication when uploading the graphics Reviewer 3: Dear author, Please, achieve next suggestions! Methods: 2. Thanks Please, improve the figure, the quality and pixeles. Reviewer 1: No Reviewer 2: No Reviewer 3: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site.
Attachment Submitted filename: Reviewer recommendations. Copyright notice. It has been added Page 20 and conclusion has been modified as- In the K. References: Magiorakos, A. There is a duplication when uploading the graphics Response: Duplicate figures have been removed. Reviewer 3: Dear author, Please, achieve next suggestions!
Thanks Response: Thank you for your valuable suggestion. Please, improve the figure, the quality and pixeles. It is suggested to place beta-lactams as a class of antibiotics Response: changed ESBL with beta-lactams accordingly. It is not necessary to capitalize Extended Response: corrected accordingly. It is not necessary to capitalize Standard Response: amended accordingly. Figure 1: Disc diffusion test for antibiotic sensitivity assessment.
Has natural resistance to this antibiotic Response: As this is a bed hospital the discs were prepared for the need of all the patients as per requirements for other growths. Why do you place an azithromycin disk? This is used in S. Typhi or Shigella Response: Herein, we aimed to assess the resistance profile of KPN isolates which were isolated from various clinical and environmental sources against common antibiotics classes.
Inpatients or outpatients Response: Resistance for inpatient and outpatient has been analyzed in the revised draft. What was the selection methodology for resistant Klebsiella? Line , The names of the bacteria must be written in italics Response: Amended accordingly. Correct typing error Response: Amended accordingly. Place a period after the word role Response: Amended accordingly. It is suggested to modify the word by site of infection Response: It has been modified accordingly.
The year of the reference should be in parentheses Response: Amended accordingly. This was stated as a probable explanation Line, Page- 17 - In this study, we considered fomites for analyzing KPN isolates as well.
Write in italics Enterobacteriacea Response: Amended accordingly. Remove the quotation mark after patients Response: Amended accordingly. Really unfortunate. Disappointed to see it has been cancelled altogether. Means we now have to miss out on seeing potential breakout performances from the likes of Dean and a potential comeback from Dawson among others,.
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