Molecular Characterization of Multi Drug Resistant (MDR) Extended Spectrum βlactamase (ESBL) Producing Klebsiella spp. from Pond Water in West Bengal (original) (raw)
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European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 2018
Carbapenem antibiotics are one of the last-resort agents against multidrug-resistant (MDR) bacteria. The occurrence of carbapenemase-producing Enterobacteriaceae (CPE) in wastewater and aquatic environments is an indication of MDR bacteria in the community. This study evaluated CPE in aquatic environments and compared them to the local hospital isolates in Sweden. Phenotypic and genotypic analyses of antibiotic resistance of environmental and clinical CPE were performed. The relatedness of the isolates and possible clonal dissemination was evaluated using phylogenetic and phyloproteomic analysis. Klebsiella oxytoca carrying carbapenemase genes (bla, bla) were isolated from wastewater and the recipient river, while K. oxytoca (bla) and Klebsiella pneumoniae (bla, bla, bla, bla) were isolated from patients at the local clinics or hospital. The K. oxytoca classified as sequence type 172 (ST172) isolated from the river was genotypically related to two clinical isolates recovered from pa...
The β-lactams are frequently used antibiotics and are essential in treatment of serious bacterial infections. The emergence of β-lactamases has been an ongoing serious therapeutic problem. To overcome this problem, scientists utilized several β-lactamases inhibitors such as clavulanic acid and tazobactam that are used in combination with antibiotics. However, pathogenic bacteria have acquired additional mechanisms of resistance such as the acquisition of extended spectrum beta-lactamases (ESBL) and carbapenemases that cannot be inhibited by current inhibitors. Carbapenem-resistant Enterobacteriaceae (CRE) has become an international health threat. In the present work, 100 clinical Klebsiella strains were isolated, identified, and their antibiotic profiles were determined by the disc diffusion method. β-lactam resistance was evaluated using phenotypic and genotypic methods. More than 50% of the Klebsiella isolates exhibited resistance to tetracyclin, ceftriaxone, (pipracillin/tazobactam), azetreonam, ofloxacin, cefepime and cefoperazone/sulbactam. Klebsiella isolates were less resistant to gentamicin (32.4%) and amikacin (14.7%). Few isolates were resistant to meropenem and imipenem (6.86%). Among the 100 clinical isolates, 50% were ESBL producers and 32% were AmpC producers. The 7 imipenem resistant isolates were carbapenemase producers. PCR showed that carbapenem resistance may be due to NDM gene that was present in 43% of the isolates and OXA gene that was found in 28% of the isolates. KPC genes were not detected in any of the isolates. Antibiotic resistance is a worldwide problem that poses a major threat to therapeutic efficacy of available antibiotics, including carbapenem. Determination of causes of resistance is essential for better treatment options.
Antibiotics
Carbapenemase-producing Enterobacteriaceae (CPE) have been heavily linked to hospital acquired infections (HAI) thereby leading to futility of antibiotics in treating infections and this have complicated public health problems. There is little knowledge about carbapenemase-producing Klebsiella spp. (CPK) in South Africa. This study aimed at determining the occurrence of CPK in different samples collected from selected environmental niches (hospitals, wastewater treatment plants, rivers, farms) in three district municipalities located in the Eastern Cape Province, South Africa. Molecular identification and characterization of the presumptive isolates were determined using polymerase chain reaction (PCR) and isolates that exhibited phenotypic carbapenem resistance were further screened for the possibility of harbouring antimicrobial resistance genes. One hundred (43%) of the 234 confirmed Klebsiella spp. isolates harboured carbapenem-resistance genes; 10 isolates harboured blaOXA-48-l...
Multiple antibiotic resistance in Klebsiella spp. and other Enterobacteriaceae isolated in Singapore
1994
A common pattern of multiple antibiotic resistance has been noted in bacteria isolated from Singaporean patients. The resistance pattern includes: ampicillin, cefuroxime, ceftazidime and other third generation cephalosporins, aztreonam, gentamicin and other aminoglycosides. The bacterial species implicated are Klebsiellas and other members of the Enterobacteriaceae. Preliminary laboratory investigation with a disk-diffusion augmentation test suggests the presence of extended-spectrum beta-lactamases. A retrospective study of laboratory blood culture records shows a rising incidence of resistance in Klebsiella spp. since 1985. Antimicrobial susceptibility results suggest a high degree of co-transfer of aminoglycoside resistance. The high frequency of this type of multiple antibiotic resistance should result in greater caution in the selection of presumptive antibiotic therapy for septicaemia, in order to avoid treatment failure and further selection of resistant strains.
Journal of thee Medical Sciences (Berkala Ilmu Kedokteran)
Background: The monitoring of antimicrobial resistance (AMR) in microorganisms that circulate in the environment is an important topic of scientific research and contributes to the development of action plans to combat the spread of multidrug-resistant (MDR) bacteria. As a synanthropic vector for multiple pathogens and a reservoir for AMR, flies can be used for surveillance. Methods: We collected 163 flies in the inner city of Berlin and examined them for extended-spectrum β-lactamase (ESBL)-producing Escherichia coli genotypically and phenotypically. Results: The prevalence of ESBL-producing E. coli in flies was 12.9%. Almost half (47.6%) of the ESBL-positive samples showed a co-resistance to ciprofloxacin. Resistance to carbapenems or colistin was not detected. The predominant ESBL-type was CTX-M-1, which is associated with wildlife, livestock, and companion animals as a potential major source of transmission of MDR E. coli to flies. Conclusions: This field study confirms the permanent presence of ESBL-producing E. coli in an urban fly population. For continuous monitoring of environmental contamination with multidrug-resistant (MDR) bacteria, flies can be used as indicators without much effort.
African Journal of Microbiology Research, 2016
The β-lactams are frequently used antibiotics and are essential in treatment of serious bacterial infections. The emergence of β-lactamases has been an ongoing serious therapeutic problem. To overcome this problem, scientists utilized several β-lactamases inhibitors such as clavulanic acid and tazobactam that are used in combination with antibiotics. However, pathogenic bacteria have acquired additional mechanisms of resistance such as the acquisition of extended spectrum beta-lactamases (ESBL) and carbapenemases that cannot be inhibited by current inhibitors. Carbapenem-resistant Enterobacteriaceae (CRE) has become an international health threat. In the present work, 100 clinical Klebsiella strains were isolated, identified, and their antibiotic profiles were determined by the disc diffusion method. β-lactam resistance was evaluated using phenotypic and genotypic methods. More than 50% of the Klebsiella isolates exhibited resistance to tetracyclin, ceftriaxone, (pipracillin/tazobactam), azetreonam, ofloxacin, cefepime and cefoperazone/sulbactam. Klebsiella isolates were less resistant to gentamicin (32.4%) and amikacin (14.7%). Few isolates were resistant to meropenem and imipenem (6.86%). Among the 100 clinical isolates, 50% were ESBL producers and 32% were AmpC producers. The 7 imipenem resistant isolates were carbapenemase producers. PCR showed that carbapenem resistance may be due to NDM gene that was present in 43% of the isolates and OXA gene that was found in 28% of the isolates. KPC genes were not detected in any of the isolates. Antibiotic resistance is a worldwide problem that poses a major threat to therapeutic efficacy of available antibiotics, including carbapenem. Determination of causes of resistance is essential for better treatment options.
Frontiers in Microbiology, 2021
This study was undertaken to evaluate the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae in selected shrimp aquaculture farms (n = 37) in Kerala, South India and to characterize the isolates using molecular tools. Overall, a low prevalence of ESBL-producers was found in the farms, most likely due to the reduced antibiotic usage in the shrimp farming sector. Out of the 261 samples (77 shrimp and 92 each of water and sediment), 14 (5.4%) tested positive for ESBL-E. coli or ESBL-K. pneumoniae. A total of 32 ESBL-E. coli and 15 ESBL- K. pneumoniae were recovered from these samples. All ESBL isolates were cefotaxime-resistant with minimal inhibitory concentration (MIC) ≥32 μg/ml. Of all isolates, 9 (28.1%) E. coli and 13 (86.7%) K. pneumoniae showed simultaneous resistance to tetracycline, ciprofloxacin, and trimethoprim-sulfamethoxazole. PCR analysis identified CTX-M group 1 (blaCTX–M–15) as the predominant ESBL genotype in bot...
Investigations of Carbapenem-resistant Klebsiella species and associated clinical considerations
2019
The use of many antibiotics to treat infections has become limited in the last decade. Enterobacteriaceae, especially Klebsiella spp., have acquired resistance to quinolones, aminoglycosides, cephalosporins and carbapenems. Resistance to β-lactams is mediated via extended-spectrum β-lactamases, AmpC type β-lactamases and carbapenemases combined with porin loss. Carbapenems are the antibiotics of last resort. The emergence of carbapenemase-producing organisms (CPOs) has led to Public Health England introducing a national toolkit to limit their spread. As part of this requirement, the Charing Cross microbiology laboratory of the Imperial College Healthcare NHS Trust revised its screening programme for the detection of CPOs. This improved the detection and isolation of CPOs, and highlighted Klebsiella spp. were more of a problem with respect to multidrug resistance than previously thought. Thirty-nine carbapenem-resistant Klebsiella strains were characterised. Phenotypic tests identified the strains as Klebsiella pneumoniae (n = 36) and Klebsiella oxytoca (n = 3). Detailed whole-genome sequence (WGS) analyses showed the K. oxytoca were Klebsiella michiganensis and one of the K. pneumoniae strains to be Klebsiella variicola subsp. variicola. The K. michiganensis strains were all of sequence type 138. They were predicted to encode the β-lactamases bla GES-5 , bla SHV-66 , bla TEM-1 , bla OXA and bla CTX-M-15 , and the 12gene operon of the kleboxymycin biosynthetic gene cluster. This gene cluster encodes for tilimycin and tilivalline, enterotoxins previously thought only to be carried by K. oxytoca strains. Incorporation of antimicrobial resistance and virulence gene data showed hypervirulent, multidrug-resistant K. pneumoniae strains encoding both aerobactin and rmpA (the regulator of mucoid phenotype) or colibactin are present in West London Hospitals. These are a cause for concern, as they have the potential to cause outbreaks that are untreatable. WGS analyses yield more accurate and comprehensive data compared with phenotypic testing, enabling exact identification of clinically important strains, detailed outbreak investigations and molecular characterisation of antibiotic resistance and virulence genes in clinical settings. Thirty-two bacteriophages were isolated from sewage water and found to infect one or more of the clinical Klebsiella isolates. Some phages with broad host ranges (i.e. they infected K. pneumoniae, K. michiganensis, K. variicola and K. grimontii strains) were identified, which may have use in clinical therapeutics against multidrug-resistant infections. These bacteriophages remain to be characterised in detail. Table 2.1: Negative controls used in this study. Control Strain Characteristics* Enterobacter cloacae M22 AmpC positive Klebsiella pneumoniae W88 ESBL positive with porin loss Enterobacter aerogenes W31 AmpC positive Enterobacter spp. X34 AmpC + ESBL positive Klebsiella pneumoniae W68 ESBL positive Escherichia coli H97 ESBL positive Klebsiella pneumoniae F44 ESBL positive Klebsiella pneumoniae H32 ESBL positive with porin loss Klebsiella pneumoniae T10 ESBL positive with porin loss Klebsiella pneumoniae T73 ESBL positive with porin loss Enterobacter spp. H88 AmpC positive with porin loss Klebsiella pneumoniae F22 CTX M ESBL positive with porin loss Enterobacter cloacae W46 AmpC positive Klebsiella pneumoniae M68 AmpC + ESBL positive Klebsiella pneumoniae X15 ESBL positive Pseudomonas aeruginosa M30 Porin loss + efflux Enterobacter cloacae F09 AmpC positive Klebsiella pneumoniae H03 ESBL positive Pseudomonas aeruginosa S89 Porin loss Enterobacter cloacae W66 ESBL + AmpC positive *AmpC, AmpC-type β-lactamase, ESBL, extended-spectrum β-lactamase; CTX M ESBL, cefotaxime-resistant ESBL. Isolated from Country Reference(s) P. aeruginosa Burn wound, clinical isolates China Wang et al. (2006); Wang et al.
RADS Journal of Biological Research & Applied Sciences
Multi drug resistance has now become a worldwide therapeutic challenge due to the widespread use of broad spectrum antibiotics. Klebsiella species have significant importance in clinical field as they cause various infections in human and are considered as potential pathogens that express antibiotic resistance through their strong enzymatic activity. Extended spectrum beta lactamases (ESBLs) are plasmid mediated enzymes produced mostly because of mutation and few other factors. These enzymes confer resistance against various β-lactam drugs including cephalosporins and monobactams. Among the genus Klebsiella, ESBLs are highly prevalent in K. pneumoniae followed by K. oxytoca. This study was conducted in Pakistan to assess the distribution of ESBL producers among Klebsiella spp., an important member of the family Enterobacteriaceae. From January 2010 to January 2012, a total of 236 gram-negative isolates were collected from different renowned microbiological laboratories. Out of the...
2019
The use of beta-lactams has tremendously increased since the discovery of antibiotics. This has led to the emergence of certain resistant genes such as Extended Spectrum Beta-Lactamase (ESBL) which confer resistance to third generation cephalosporins. The objective of this study was to determine the prevalence of Extended Spectrum Beta Lactamase (ESBL) genes, Carbapenem resistance genes (blaKPC, blaOXA and IMP) and outer membrane porins genes (OMP-35, OMP-36, and OMP-36N) from different hospitals and laboratories in Ikeja-Lagos, Nigeria. A total of 177 bacterial isolates were collected between May 2017 to July 2017 from patients with urinary tract infections (UTI) and gastroenteritis. They were identified biochemically and investigated for ESBL and Carbapenemase production using phenotypic Double Disk Synergy Test (DDST) and Modified Hodges’ Test respectively. Antibiotics susceptibility profile was also investigated. Multiplex PCR was used to detect the genes responsible for the res...