Browsing by Author "Debadatta Dhar Chanda"

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Co-carriage of blaKPC-2 and blaNDM-1 in clinical isolates of Pseudomonas aeruginosa associated with hospital infections from India
(Public Library of Science, 2015) Deepjyoti Paul; Debadatta Dhar Chanda; Anand Prakash Maurya; Shweta Mishra; Atanu Chakravarty; Gauri Dutt Sharma; Amitabha Bhattacharjee
Global spread of KPC poses to be a serious threat complicating treatment options in hospital settings. The present study investigates the genetic environment of blaKPC-2 among clinical isolates of Pseudomonas aeruginosa from a tertiary referral hospital of India. The study isolates were collected from different wards and clinics of Silchar Medical College and Hospital, India, from 2012-2013. The presence of blaKPC was confirmed by genotypic characterization followed by sequencing. Cloning of the blaKPC-2 gene was performed and the genetic environment of this gene was characterized as well. Transferability of the resistance gene was determined by transformation assay and Southern hybridization. Additionally restriction mapping was also carried out. Two isolates of P. aeruginosa were found to harbor blaKPC-2 were resistant towards aminoglycosides, quinolone and β-lactam-β-lactamase inhibitor combination. In both the isolates, the resistance determinant was associated with class 1 integron and horizontally transferable. Both the isolates were co-harboring blaNDM-1. The first detection of this integron mediated blaKPC-2 coexisting with blaNDM-1 in P. aeruginosa from India is worrisome, and further investigation is required to track the gene cassette mediated blaKPC-2 in terms of infection control and to prevent the spread of this gene in hospitals as well as in the community. © 2015 Paul et al. 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.
Diverse Genetic Array of blaCTXM-15 in Escherichia coli: A Single-Center Study from India
(Mary Ann Liebert Inc., 2016) Anand Prakash Maurya; Shweta Mishra; Anupam Das Talukdar; Debadatta Dhar Chanda; Atanu Chakravarty; Amitabha Bhattacharjee
CTX-M-15 is a chief contributor for expanded-spectrum cephalosporin and monobactam resistance in India, complicating treatment options. In this study, we have investigated genetic context of CTX-M-15 in Escherichia coli and their transmission dynamics in a tertiary referral hospital of India. A total of 198 isolates were collected, of which 66 were harboring blaCTXM-15. Among them, 14 isolates were carrying a single CTX-M-15 gene and 52 were harboring multiple extended-spectrum β-lactamase genes along with blaCTX-M-15. The resistance gene was flanked by tnpA, ISEcp1, IS26, and ORF477 in 10 different arrangements. The resistance determinant was horizontally transferable through F, W, I1, and P Inc types of plasmids. Restriction mapping of plasmids showed a variable band pattern even within the same Inc types. Minimum inhibitory concentration was found above the breakpoint level against expanded-spectrum cephalosporins and monobactam while susceptible against carbapenems. blaCTX-M-15 was highly stable and sustained in the cell after 115 serial passages. In pulse-field gel electrophoresis, eight pulsotypes of E. Coli were found to be responsible for the spread of blaCTX-M-15 in the tertiary referral center. We conclude that the presence of CTX-M-15 in the heterogeneous group of E. Coli is highly alarming in terms of infection control and it may require regular monitoring, so as to formulate appropriate antibiotic policy to stop the spread of this resistance determinant. © Copyright 2016, Mary Ann Liebert, Inc. 2016.
Use of fluorescence foldscope as an effective tool for detection of biofilm formation in Pseudomonas aeruginosa
(Wolters Kluwer Medknow Publications, 2020) Chandrayee Deshamukhya; Bhaskar Das; Shiela Chetri; Deepjyoti Paul; Debadatta Dhar Chanda; Tuhina Banerjee; Amitabha Bhattacharjee
Purpose: Pseudomonas aeruginosa is an opportunistic pathogen with biofilm-forming ability, by the virtue of which they can evade the immune response and antimicrobial chemotherapy. Several methods have been designed for the detection of biofilms but require sophisticated instrumentation and expertise. The present study, therefore, used an improvised device, 'fluorescence foldscope' which is an origami-based fluorescence microscope as an easy and effective tool to detect biofilm formation. Methodology: Three representatives of P. aeruginosa of clinical origin were taken for the study along with two reference strains PA01 and ATCC27853. The strains were cultured in Luria Bertani (LB) broth with and without carbapenem (imipenem and meropenem) and cephalosporin (ceftazidime, cefotaxime and ceftriaxone) pressure, respectively. The cultures were diluted to 1:100 in LB; seeded with sterile glass slides at 90° angle and incubated for 5 consecutive days. The slides were observed with fluorescence foldscope. Results: Fluorescence emission was observed in two test isolates CD1 and CD2 at 48 and 72 h, respectively, whereas no fluorescence was observed in CD3. The fluorescence observed in the isolates was not affected by 2 μg/ml carbapenem pressure, while with 2 μg/ml ceftazidime stress, a change in fluorescence was observed in CD2 in comparison to the fluorescence observed under normal growth condition. Conclusion: Fluorescence foldscopy is an effective and reliable tool for the detection of biofilm formation in clinical isolates of P. aeruginosa under different laboratory conditions. Biofilm-forming P. aeruginosa worsens the medical condition and is difficult to eradicate. The present study came up with an effective and reliable tool for the detection of biofilm formation in clinical isolates of P. aeruginosa. © 2020 Wolters Kluwer Medknow Publications. All rights reserved.