ESBL Producers in Gram Negative Isolates from Clinical Samples

Resistance to beta lactam antibiotics is the most common cause for beta-lactamase production. Increasing number of extended spectrum beta-lactamase (ESBL) producers has reduced the treatment options which resulted in emergence of multidrug resistant strains, treatment failure and hence increased mortality. To detect phenotypically, ESBL producers in Gram negative isolates from different samples and to know their susceptibility pattern. A retrospective study of Gram negative isolates was conducted. Total of 521 isolates were isolated from various samples. They were processed and identified by standard procedures. The antibiotic susceptibility testing was performed by Kirby- Bauer disc diffusion method using CLSI guidelines. ESBL was detected by combination disk test. A total of 521 Gram negative isolates were isolated which included E. coli, Klebsiella pneumoniae, Citrobacter spp., Enterobacter spp., Proteus spp. and Acinetobacter spp. Pseudomonas aeruginosa. Of 521 isolates tested, ESBL was detected in 329 (63.1%) isolates. These isolates showed maximum susceptibility to piperacillin- tazobactam (86%) followed by imipenem (78.4%), amikacin (63.5%), cotrimoxazole (54.4%), ciprofloxacin (51%), amoxi-clav (44.9%), cefepime (44.1%), gentamicin (38.9%), cefoxitin (34.9%) and ampicillin (19.1%). ESBL producers which are resistant to beta lactam antibiotics have become a major problem. Detection of these beta-lactamase enzymes by simple disk method and its reporting will help clinicians in prescribing proper antibiotics.

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PREVALENCE OF CEPHALOSPORIN-RESISTANT GRAM-NEGATIVE BACILLI FROM CLINICAL SAMPLES

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2016.v9s2.13456 ◽

2016 ◽

pp. 176

Author(s):

Kavi Aniis ◽

Rajamanikandan Kcp ◽

Arvind Prasanth D

Keyword(s):

Clinical Samples ◽

Beta Lactamase ◽

Bacterial Isolates ◽

Beta Lactam ◽

Gram Negative ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Lactam Ring ◽

Esbl Producers

ABSTRACT Objective: Beta-lactams are the group of antibiotics that contain a ring called as “beta-lactam ring,” which is responsible for the antibacterial activity. The presence of resistance among Gram-negative organisms is due to the production of beta-lactamases enzymes that hydrolysis the beta-lactam ring thereby conferring resistance to the organism. This study is undertaken to determine the prevalence of extended-spectrum beta-lactamase (ESBL) producing Gram-negative organism from clinical samples. Methods: A total of 112 clinical samples were taken for this study. The combined disc synergistic test (CDST) was used for the phenotypic detection of ESBL producers from the clinical samples. The genotypic identification of ESBL producers was carried out by alkaline lysis method by isolation of plasmid DNA. Result: A total of 87 bacterial isolates were isolated and identified. Among them, Klebsiella (41%) was the predominant organism followed by Escherichia coli (33%), Proteus (10%), Pseudomonas (10%), and Serratia (6%). Among the various bacterial isolates, Klebsiella showed a higher percentage of resistance. The CDST showed that 8 isolates of Klebsiella, 3 isolates of E. coli, and 1 isolate of Pseudomonas were found to be ESBL producers. The genotypic confirmation showed that the two bacterial isolates, namely, Klebsiella and E. coli were found to possess temoniera (TEM) gene which was the 400-500 bp conferring resistance to the antibiotics. Conclusion: The results of this study suggest that early detection of ESBL producing Gram-negative organism is a very important step in planning the therapy of patient in Hospitals. CDST continues to be a good indicator in the detection of ESBL producers. Keywords: Beta-lactamases, Gram-negative bacilli, Extended-spectrum beta-lactamase, Resistance, Combined disc synergistic test.

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Coproduction of Extended Spectrum, AmpC and Metallo β- Lactamases in Pseudomonas aeruginosa Isolates from a Super Speciality Center

International Journal of Current Microbiology and Applied Sciences ◽

10.20546/ijcmas.2021.1010.020 ◽

2021 ◽

Vol 10 (10) ◽

pp. 176-184

Author(s):

Ashna Bhasin Poonam Loomba ◽

Abha Sharma Bibhabati Mishra ◽

Ashish Bajaj

Keyword(s):

Pseudomonas Aeruginosa ◽

Treatment Options ◽

Multidrug Resistant ◽

Clinical Isolates ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum

Pseudomonas aeruginosa (P. aeruginosa) is one of the leading causes of hospital as well as community acquired infections. They’re strenuous to treat as most of isolates exhibit various degrees of beta- lactamase mediated resistance to majority of the beta-lactam antibiotics. Single isolate can express multiple β- lactamase enzymes, further limiting the treatment options. Therefore, this study was outlined to research the coexistence of various beta-lactamase enzymes in clinical isolates of P. aeruginosa. The aim of the study was to detect the co-prevalence of Extended Spectrum Beta lactmases (ESBL), AmpC and Metallo β-Lactamases (MBL) in Pseudomonas aeruginosa isolates from a superspeciality center. Fifty clinical isolates of P. aeruginosa were tested for the presence of AmpC beta-lactamase, extended spectrum beta- lactamase (ESBL) and metallo beta-lactamase (MBL) enzyme. Discernment of AmpC beta-lactamase was performed by disk antagonism while ESBL detection was done by the combined disk diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guidelines and MBL were detected by the Imipenem EDTA disk potentiation test. Eleven of 50 (22%) isolates were confirmed to be positive for AmpC and Extended spectrum beta lactamases. Co-production of AmpC along side ESBL and MBL was reported in 12 % isolates. The study shows the high prevalence of multidrug resistant P. aeruginosa producing beta-lactamase enzymes of diverse mechanisms. Consequently, formulation of a correct antibiotic policy and taking measures to restrict the indiscriminative use of cephalosporins and carbapenems should be taken to mitigate the emergence of this multiple beta-lactamase producing pathogens.

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Prevalence and Antibiogram of Extended-Spectrum Beta-Lactamase Producing Gram-negative Bacteria Isolated from Septicemic Neonates in a Tertiary Care Hospital

KYAMC Journal ◽

10.3329/kyamcj.v11i4.51991 ◽

2021 ◽

Vol 11 (4) ◽

pp. 171-175

Author(s):

Tania Rahman ◽

Momtaz Begum ◽

Sharmeen Sultana ◽

SM Shamsuzzaman

Keyword(s):

Antimicrobial Susceptibility ◽

Diffusion Method ◽

Gram Negative Bacteria ◽

Care Hospital ◽

Beta Lactamase ◽

Blood Samples ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Esbl Producers

Background: In recent years, Extended-spectrum beta-lactamase (ESBL) producing microorganisms have complicated treatment of infections due to resistance of ESBL producing strains to a wide range of antimicrobials. Objective: Target of this study was to determine the prevalence of ESBL producing gramnegative bacteria in neonatal sepsis cases and to reveal the antimicrobial susceptibility pattern of those isolated ESBL producers. Materials and Methods: This cross sectional study was carried out in Dhaka Medical College Hospital (DMCH) over a period of 12 months from January to December in 2016. Following isolation and identification of gram-negative bacteria from blood samples of suspected septicemic neonates, antimicrobial susceptibility test was performed by Kirby Bauer disk-diffusion method and ESBL producers were detected by Double Disk Synergy (DDS) test. Results: Among 52 Gram-negative bacteria isolated from 106 blood samples, 34.61% ESBL producers were detected and Enterobacter spp. (45%) was predominant followed by Klebsiella pneumoniae (33.33%). None of the ESBL producers was resistant to colistin and tigecycline. All ESBL producing Acinetobacter baumannii, 77.78% and 66.67% of ESBL producing Enterobacter spp and Klebsiella spp. respectively showed resistance to meropenem. All ESBL producers were resistant to piperacillintazobactam. Conclusion: Appropriate measures should be taken to prevent the spread of ESBL producing strains by combining strategies for infection prevention, control and rational use of antibiotics. KYAMC Journal Vol. 11, No.-4, January 2021, Page 171-175

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Presence of different beta-lactamase classes among clinical isolates of Pseudomonas aeruginosa expressing AmpC beta-lactamase enzyme

The Journal of Infection in Developing Countries ◽

10.3855/jidc.497 ◽

2010 ◽

Vol 4 (04) ◽

pp. 239-242 ◽

Cited By ~ 16

Author(s):

Supriya Upadhyay ◽

Malay Ranjan Sen ◽

Amitabha Bhattacharjee

Keyword(s):

Pseudomonas Aeruginosa ◽

Treatment Options ◽

Clinical Isolates ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Enzyme Detection

Introduction: Infections caused by Pseudomonas aeruginosa are difficult to treat as the majority of isolates exhibit varying degrees of beta-lactamase mediated resistance to most of the beta-lactam antibiotics. It is also not unusual to find a single isolate that expresses multiple β-lactamase enzymes, further complicating the treatment options. Thus the present study was designed to investigate the coexistence of different beta-lactamase enzymes in clinical isolates of P. aeruginosa. Methodology: A total of 202 clinical isolates of P. aeruginosa were tested for the presence of AmpC beta-lactamase, extended spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) enzyme. Detection of AmpC beta-lactamase was performed by disk antagonism test and a modified three-dimensional method, whereas detection of ESBL was done by the combined disk diffusion method per Clinical and Laboratory Standards Institute (CLSI) guidelines and MBL were detected by the Imipenem EDTA disk potentiation test. Results: A total of 120 (59.4%) isolates were confirmed to be positive for AmpC beta-lactamase. Among them, 14 strains (7%) were inducible AmpC producers. Co-production of AmpC along with extended spectrum beta-lactamase and metallo beta-lactamase was reported in 3.3% and 46.6% isolates respectively. Conclusion: The study emphasizes the high prevalence of multidrug resistant P. aeruginosa producing beta-lactamase enzymes of diverse mechanisms. Thus proper antibiotic policy and measures to restrict the indiscriminative use of cephalosporins and carbapenems should be taken to minimize the emergence of this multiple beta-lactamase producing pathogens.

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Molecular Detection of Carbapenem Resistant Gram Negative Bacterial Isolates from Dogs

Indian Journal of Animal Research ◽

10.18805/ijar.b-4297 ◽

2021 ◽

Author(s):

Surya Sankar ◽

Thresia . ◽

Anu Bosewell ◽

M. Mini

Keyword(s):

Companion Animals ◽

Multidrug Resistant ◽

Beta Lactam ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Beta Lactam Antibiotics ◽

Carbapenem Resistant ◽

Carbapenemase Gene ◽

Line Of Therapy ◽

Encoding Genes

Background: Carbapenems are beta-lactam antibiotics that are considered as the last line of therapy against multidrug resistant extended spectrum beta-lactamase. The resistance to carbapenems predominantly through carbapenemase is one of the most important emerging health problems worldwide in the therapy of clinical infections. The objective of the present study is to determine the presence of carbapenemase encoding genes among Gram- negative bacterial spp. associated with clinical infections in dogs. Methods: 30 Escherichia coli, 11 Klebsiella pneumoniae and three Pseudomonas aeruginosa isolated from urine, swabs from lesional skin and anterior vagina of dogs presented with different clinical ailments formed the samples for the study. Polymerase chain reaction was carried out to detect the presence of carbapenemase encoding genes viz., KPC, NDM, OXA, VIM and IMP among the isolates.Result: Out of the 44 Gram- negative isolates tested, 28 (76.3%) were positive for at least one tested carbapenemase gene. The highest frequency of carbapenemase recorded was for NDM followed by OXA-181, KPC, OXA-48 and VIM. Our study identified a high prevalence of carbapenemases among companion animals like dogs which could act as potential source of transmission of these resistance bacteria or their genomes to humans.

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Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-ProducingEscherichia coliamong Uropathogens of Pediatrics in North of Iran

BioMed Research International ◽

10.1155/2015/309478 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 24

Author(s):

Mohammad Sadegh Rezai ◽

Ebrahim Salehifar ◽

Alireza Rafiei ◽

Taimour Langaee ◽

Mohammadreza Rafati ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Multidrug Resistant ◽

Beta Lactamase ◽

Beta Lactam ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases ◽

North Of Iran

Escherichia coliremains as one of the most important bacteria causing infections in pediatrics and producing extended-spectrum beta-lactamases (ESBLs) making them resistant to beta-lactam antibiotics. In this study we aimed to genotype ESBL-producingE. coliisolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of theE. coliisolates were initially considered ESBL producing based on their MIC results. These isolates were then tested by polymerase chain reaction (PCR) for the presence or absence ofCTX,TEM,SHV,GES, andVEBbeta-lactamase genes. About 30.5% of isolatedE. coliwas ESBL-producing strain. TheTEMgene was the most prevalent (49%) followed bySHV(44%),CTX(28%),VEB(8%), andGES(0%) genes. The ESBL-producingE. coliisolates were susceptible to carbapenems (66%) and amikacin (58%) and showed high resistance to cefixime (99%), colistin (82%), and ciprofloxacin (76%). In conclusion, carbapenems were the most effective antibiotics against ESBl-producingE. coliin urinary tract infection in North of Iran. The most prevalent gene is the TEM-type, but the other resistant genes and their antimicrobial resistance are on the rise.

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First Global Report of Plasmid-Mediated mcr-1 and Extended-Spectrum Beta-Lactamase-Producing Escherichia coli from Sheep in Portugal

Antibiotics ◽

10.3390/antibiotics10111403 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1403

Author(s):

Josman Dantas Palmeira ◽

Marisa Haenni ◽

Jean-Yves Madec ◽

Helena Maria Neto Ferreira

Keyword(s):

One Health ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Colistin Resistance ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Extended Spectrum

Resistances to extended-spectrum cephalosporins (ESC) and colistin are One Health issues since genes encoding these resistances can be transmitted between all sectors of the One Health concept, i.e., human, animal, and the environment. Among food-producing animals, sheep farming has long been overlooked. To fill in this knowledge gap, we looked for ESC- and colistin resistance in 21 faecal samples collected from sheep in one farm in the south of Portugal. ESC-resistant isolates were selected on MacConkey agar plates supplemented with cefotaxime. Susceptibility testing was performed by the disk-diffusion method according to CLSI, while colistin MIC was determined by broth microdilution. ESC- and colistin-resistance genes were identified by PCR, and the clonality of all isolates was assessed by XbaI-PFGE. The replicon content was determined by PCR according to the PCR-based replicon typing (PBRT) scheme. Sixty-two non-duplicate ESC-resistant E. coli isolates were identified, which all presented an extended-spectrum beta-lactamase (ESBL) phenotype, mostly due to the presence of CTX-M genes. One CTX-M-1-producing E. coli was concomitantly colistin-resistant and presented the plasmid-mediated mcr-1 gene. Nearly all isolates showed associated resistances to non-beta-lactam antibiotics, which could act as co-selectors, even in the absence of beta-lactam use. The results showed a high proportion of ESBL-producing E. coli in sheep faeces. Their dissemination was very dynamic, with the spread of successful clones between animals, but also a large diversity of clones and plasmids, sometimes residing in the same animal. This study highlights the need for global surveillance in all food-producing sectors, in order to avoid the dissemination of genes conferring resistance to last-resort antibiotics in human medicine.

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Clinical Relevance of Antibiotic Susceptibility Profiles for Screening Gram-negative Microorganisms Resistant to Beta-Lactam Antibiotics

Microorganisms ◽

10.3390/microorganisms8101555 ◽

2020 ◽

Vol 8 (10) ◽

pp. 1555 ◽

Cited By ~ 1

Author(s):

Francisco Montiel-Riquelme ◽

Elisabeth Calatrava-Hernández ◽

Miguel Gutiérrez-Soto ◽

Manuela Expósito-Ruiz ◽

José María Navarro-Marí ◽

...

Keyword(s):

Resistant Bacteria ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Beta Lactam ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Beta Lactam Antibiotics ◽

Clinical Laboratories ◽

Phenotypic Methods ◽

Susceptibility Profiles

The increasing resistance to antibiotics is compromising the empirical treatment of infections caused by resistant bacteria. Rapid, efficient, and clinically applicable phenotypic methods are needed for their detection. This study examines the phenotypic behavior of β-lactam-resistant Gram-negative bacteria grown on ChromID ESBL medium with ertapenem, cefoxitin, and cefepime disks, reports on the coloration of colonies, and establishes a halo diameter breakpoint for the detection of carbapenemase-producing bacteria. We studied 186 β-lactam-resistant Gram-negative microorganisms (77 with extended spectrum beta lactamase (ESBL), 97 with carbapenemases, and 12 with AmpC β-lactamases (AmpC)). Susceptibility profiles of Gram-negative bacteria that produced ESBL, AmpC, and carbapenemases were similar to the expected profiles, with some differences in the response to cefepime of ESBL-producing microorganisms. Coloration values did not differ from those described by the manufacturer of ChromID ESBL medium. In the screening of carbapenemase production, inhibition halo diameter breakpoints for antibiotic resistance were 18 mm for Enterobacterales and ertapenem, 18 mm for Pseudomonas and cefepime, and 16 mm for Acinetobacter baumannii and cefepime. This innovative phenotypic approach is highly relevant to clinical laboratories, combining susceptibility profiles with detection by coloration of high-priority resistant microorganisms such as carbapenemase-producing A. baumannii, carbapenemase-producing Pseudomonas spp., and ESBL and/or carbapenemase-producing Enterobacterales.

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Extended Spectrum Beta Lactamase (ESBL) Producing Klebsiella pneumoniae , a therapeutic challenge.

Medical Journal of Shree Birendra Hospital ◽

10.3126/mjsbh.v13i1.12996 ◽

2015 ◽

Vol 13 (1) ◽

pp. 22-25

Author(s):

Raina Chaudhary ◽

Sabita Bhatt Bhatt ◽

Eva Piya

Keyword(s):

Klebsiella Pneumoniae ◽

Multidrug Resistant ◽

Diffusion Method ◽

Clinical Samples ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Esbl Producer ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Sensitivity Pattern

Introduction: Klebsiella pneumoniae is one of the most common Gram negative bacteria encountered byclinicians worldwide as a cause of infections in human. Most of the infections are acquired in hospital settingtherefore, it is reported to be the amongst the 10 most common nosocomial pathogen in various studies. Nowadays,Klebsiella pneumoniae infections are complicated by increase in Extended Spectrum Beta Lactamase (ESBL)producing isolates. Therefore, this study is being conducted with the objective to fi nd out the prevalence ofESBL producing Klebsiella pneumoniaein various clinical samples and to fi nd out there sensitivity pattern.Methods: A total of 100 Klebsiella pneumoniae were isolated from various samples during the period of April2013 to November 2013 in Microbiology Unit of Shree Birendra Hospital. All the isolates were identifi ed withtheir sensitivity pattern according to standard methodology. Combination disc diffusion method was followedfor identifi cation of ESBL.Results: Out of total 100 isolates of Klebsiella pneumoniae21% were ESBL producer.ESBL producer isolatesshowed 100% sensitivity to Imepenem followed by Amikacin 57.1% and Chloramphenicol 47.6%. All theESBL isolates were resistant to both Cefotaxime and Ceftazidime.Conclusions: ESBL producer Klebsiella pneumoniae isolates were multidrug resistant. Continuous surveillanceand timely intervention with discouraging the use of cephalosporin group of antibiotics is mandatory.doi: http://dx.doi.org/10.3126/mjsbh.v13i1.12996

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Phenotypic characterization of beta-lactamases producing Gram-negative bacteria in a tertiary hospital, Nepal

Nepal Journal of Biotechnology ◽

10.3126/njb.v7i1.26954 ◽

2019 ◽

Vol 7 (1) ◽

pp. 74-81

Author(s):

Elina Maharjan ◽

Pooja Shakya ◽

Balkrishna Bhattachan ◽

Bharat Prasad Baral ◽

Dhiraj Shrestha

Keyword(s):

Tertiary Hospital ◽

Diffusion Method ◽

Phenotypic Characterization ◽

Clinical Samples ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Gram Negative ◽

Beta Lactamases ◽

Disk Method ◽

Esbl Producers

Infections caused by beta-lactamases producing Gram-negative bacteria are increasing, thus posing a challenge to the management of such infections. The surveillance data of such bacteria is limited in Nepal so this study aimed to detect the beta-lactamase producing Gram-negative bacteria in a tertiary setting. A total of 604 clinical samples, including urine, blood, sputum and body fluids, were cultured and identified by the routine standard laboratory protocols. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method following Clinical and Laboratory Standard Institute guidelines (2014). Extended-spectrum beta-lactamases (ESBL) producers were identified by combined disk method and metallo-beta-lactamases (MBL) producers were identified by Imipenem- EDTA combined disk method. Out of 604 samples, 282 (46.7%) samples showed significant growth, of which 229 (81.2%) were Gram-negative bacteria. Of 229 Gram-negative bacteria, 200 (87.3%) were multidrug resistant, 67 (29.3%) were ESBL producers and 16 (7.0%) were MBL producers. Klebsiella pneumoniae were among higher ESBL producers and Pseudomonas aeruginosa were among higher MBL producers. The findings suggest higher antibacterial resistance among Gram-negative bacteria with the added burden of beta-lactamase production. Imipenem was effective against 125 of 229 Gram-negative bacteria tested. Thus, imipenem can be the drug of choice for empirical management. The higher multidrug resistance and higher beta-lactamases production among Gram-negative bacteria warrant the continuous monitoring, surveillance, early detection, and infection control practices of such bacteria

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