scholarly journals Detection of prevalence of metallo-beta lactamases in clinical isolates of imipenem resistant Pseudomonas aeruginosa from neonatal septicaemia cases in a tertiary hospital in Odisha, India

2017 ◽  
Vol 5 (1) ◽  
pp. 61 ◽  
Author(s):  
Surya Narayan Mishra ◽  
Seba Ranjan Biswal ◽  
Basanta Kumar Behera ◽  
Dipti Pattnaik
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
Strict Adherence ◽  
Neonatal Septicemia ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Pseudomonas Spp ◽  
Neonatal Septicaemia ◽  
Synergy Test

Background: Pseudomonas aeruginosa is a clinically troublesome gram-negative pathogen that causes both opportunistic infections and nosocomial outbreaks. Metallo beta lactamase have recently emerged as a worrisome resistance mechanism. Carbapenems had been the drug of choice for the infections caused by most penicillin- or cephalosporin-resistant gram-negative bacteria due to its broad-spectrum activity and stability to hydrolysis by most beta-lactamases. This does not hold good anymore due to rapid uprise of MBL producing strains. The current research covered 163 hospitalized cases of neonatal septicaemia from which Pseudomonas aeruginosa is isolated in the Paediatric Department of KIMS, Bhubaneswar. The study aimed at detecting the prevalence of metallo-beta lactamases in clinical isolates of imipenem resistant Pseudomonas aeruginosa from neonatal septicemia cases and to establish the antibiogram of Imipenem-resistant P. aeruginosa these cases. Methods: Clinical samples obtained from suspected cases of neonatal septicemia were first cultured by conventional methods and then identification was done by VITEK-2 instrument. Metallo beta lactamase (MBL) production was done by combined disc synergy test (CDST) using imipenem and EDTA (CDST-IPM) and double disc synergy test (DDST) using IPM and EDTA (DDST-IPM). Results: Among 1510 processed clinical specimens from cases of neonatal septicaemia; 637 (42.18%) showed positive growth of various clinically significant pathogens. Out of them in 163 (25.58%) cases Pseudomonas spp. was isolated. Of these, a total of 95 (58.28%) Pseudomonas spp. was found resistant to imipenem. All imipenem-resistant Pseudomonas isolates were positive for MBL by CDST imipenem-EDTA (CDST-IPM) method, whereas 89 (93.68%) were positive by DDST-IPM method, respectively. Pseudomonas aeruginosa was mostly isolated from endotracheal tube aspirate (57.89%) followed by pus (56.41%). Out of the 95 cases of MBL-producing Pseudomonas; 46 (48.42%) isolates showed the maximum susceptibility to piperacillin-tazobactam combination. All MBL-producing Pseudomonas isolates were resistant to ceftriaxone.Conclusions: MBL-producing Pseudomonas is found to be highly prevalent in our hospital, which is one of the major causes of multidrug resistance and need regular surveillance and strict adherence to a robust antibiotic policy.

scholarly journals Metallo-beta-lactamase producing Pseudomonas aeruginosa in neonatal septicemia

2010 ◽  
Vol 2 (01) ◽  
pp. 014-016 ◽  
Author(s):  
Madhu Sharma ◽  
Sarita Yadav ◽  
Uma Chaudhary
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Neonatal Sepsis ◽  
Antimicrobial Agents ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
Neonatal Septicemia ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Gram Negative Organisms

ABSTRACTGram-negative bacilli are important agents causing neonatal sepsis. The organisms isolated are often resistant to multiple antimicrobials specially which are metallo-beta-lactamases (MβL) producers. Therefore, the present study was conducted with the objective to examine the incidence of MβL producing strains in multidrug resistant (MDR) Pseudomonas aeruginosa from cases of neonatal sepsis. Between January-December 2006, 1994 cases of neonatal sepsis were investigated. The isolates obtained were identified and tested for susceptibility to various antimicrobial agents. The multidrug resistant P. aeruginosa isolates were screened for the presence of MβL by imipenem-EDTA disc method. Five hundred and ninety three (29.73%) isolates were obtained from culture of neonates. Most frequent offender was P. aeruginosa (48.2%). There was an overall predominance of gram-negative organisms. MβL production was seen in 69.5% of imipenem-resistant P. aeruginosa isolates. MβL producing P. aeruginosa is an emerging threat in neonatal septicemia and a cause of concern for physicians treating such infections.

scholarly journals Investigation of Metallo-Beta-Lactamases in Carbapenem Resistant Pseudomonas aeruginosa Strains by Phenotypic and Genotypic Methods

2020 ◽  
Vol 25 (3) ◽  
pp. 301-307
Author(s):  
M. Duygu Aksoy ◽  
H. Murat Tuğrul
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Ethylenediaminetetraacetic Acid ◽  
Carbapenem Resistance ◽  
Beta Lactamase ◽  
Bacterial Strains ◽  
Beta Lactamases ◽  
Carbapenem Resistant ◽  
Synergy Test ◽  
Phenotypic Tests ◽  
Positive Controls

Introduction: Carbapenem resistant Pseudomonas aeruginosa strains cause serious problems in treatment. A large number of identified metallo-beta-lactamase (MBL) enzymes produced by P. aeruginosa are one of the most important mechanisms in resistance to carbapenems. MBL genes are located on the chromosome or plasmid, and they can easily spread between different bacterial strains. The activities of these enzymes are zinc-dependent, and they are inhibited by ethylenediaminetetraacetic acid (EDTA). Therefore, this advantage is used in MBL identification tests. In this study, it was aimed to determine MBL among P. aeruginosa strains. Materials and Methods: MBL existence was investigated in 35 P. aeruginosa strains accepted to be mildly susceptible/resistant to any of the carbapenem group of antibiotics through phenotypic and genotypic methods. Phenotypic tests were performed as double disk synergy test (DDST), combined disk diffusion tests (CDDT) by using 0.1 M and 0.5 M EDTA, MBL E-test, and modified Hodge test (MHT). blaIMP, blaVIM, blaGIM, blaSIM, blaSPM genes and blaNDM gene were investigated by multiplex polimerase chain reaction (PCR) and PCR, respectively. Escherichia coli ATCC 25922 and P. aeruginosa ATCC 27853 standard bacteria were used in tests. VIM-1, VIM-2, IMP-13, SPM-1, NDM-1 type MBL-producing P. aeruginosa strains were used as positive controls. Results: Among the carbapenems resistant P. aeruginosa isolates, positivity of MBL was found as 54.2% by MBL E-test, 42.8% by DDST, 94.2% and 37.1% by CDDT method using 0.5 M and 0.1 M EDTA, respectively. Modified Hodge test and genotypic method did not detect MBL. Conclusion: In order to correctly evaluate the results of the phenotypic method, the investigation of resistance genes by molecular methods is also required. The most common metallo-beta-lactamase enzymes responsible for resistance to carbapenem in Pseudomonas were not observed. It was thought that different mechanisms might be responsible for the identified carbapenem resistance.

scholarly journals In Vitro Activity of the Ultra-Broad-Spectrum Beta-lactamase Inhibitor QPX7728 in Combination with Multiple Beta-Lactam Antibiotics against Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Olga Lomovskaya ◽  
Debora Rubio-Aparicio ◽  
Kirk Nelson ◽  
Dongxu Sun ◽  
Ruslan Tsivkovski ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Resistance Mechanisms ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
In Vitro Activity ◽  
Beta Lactam ◽  
Beta Lactams ◽  
Beta Lactamases ◽  
Lactamase Inhibitor

QPX7728 is an ultra-broad-spectrum beta-lactamase inhibitor with potent inhibition of key serine and metallo beta-lactamases. QPX7728 enhances the potency of multiple beta-lactams in beta-lactamase producing Enterobacterales and Acinetobacter spp. In this study we evaluated the in vitro activity of QPX7728 (8 μg/ml) combined with multiple beta-lactams against clinical isolates of Pseudomonas aeruginosa with varying beta-lactam resistance mechanisms. Seven-hundred-ninety clinical isolates were included in this study; 500 isolates, termed a “representative panel”, were selected to be representative the MIC distribution of meropenem (MEM), ceftazidime-avibactam (CAZ-AVI), and ceftolozane-tazobactam (TOL-TAZ) resistance for clinical isolates according to 2017 SENTRY surveillance data (representative panel). An additional 290 selected isolates (“challenge panel”), that were either non-susceptible to MEM or were resistant to TOL-TAZ or CAZ-AVI were also tested; 61 strains carried metallo beta-lactamases (MBLs), 211 strains were defective in the carbapenem porin OprD and 185 strains had the MexAB-OprM efflux pump overproduced based on a phenotypic test. Against the representative panel, susceptibility for all QPX7728/beta-lactam combinations was >90%. For the challenge panel, QPX-ceftolozane (TOL) was the most active combination (78.6% susceptible) followed by equipotent QPX-piperacillin (PIP) and QPX-cefepime (FEP), restoring susceptibility in 70.3% of strains (CLSI breakpoints for the beta-lactam compound alone). For MBL-negative strains, QPX-TOL and QPX-FEP restored the MIC values to susceptibility rates in ∼90% and ∼80% of strains, respectively, vs 68-70% for QPX-MEM and QPX-PIP and 63-65% for TOL-TAZ and CAZ-AVI. For MBL-positive strains, QPX-PIP restored the MIC to susceptibility values for ∼70% of strains vs 2-40% for other combinations. Increased efflux and impaired OprD had varying effect on QPX7728 combination depending on the partner beta-lactam tested. QPX7728 enhanced the potency of multiple beta-lactams against P. aeruginosa, with varying results according to the beta-lactamase production and other intrinsic resistance mechanisms.

scholarly journals Detection of metallo beta lactamase production in imipenem resistant gram negative bacilli non fermenters isolated in a Tertiary Care Hospital.

2019 ◽  
Vol 26 (12) ◽  
pp. 2080-2084
Author(s):  
Ayesha Sajjad ◽  
Muna Malik ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Fareeha Imran ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Tertiary Care ◽  
Care Hospital ◽  
Beta Lactamase ◽  
Culture Methods ◽  
Cross Sectional ◽  
Gram Negative ◽  
Synergy Test ◽  
Pseudomonas Luteola

Objectives: Metallo-beta lactamase (MBL) producing non fermenter Gram negative bacilli is an emerging warning and cause of worry as they have established as one of the most feared resistance mechanisms and are the foremost cause of nosocomial infections worldwide. Carbapenem, including Imipenem, Meropenem and Doripenem are often used as a last remedy for treatment of infections caused by Pseudomonas aeruginosa, Acinetobacter and other Gram-negative. The present study was designed to explore the distribution of imipenem resistant non-fermenter Gram-negative bacilli isolates in different age groups. Study Design: Cross sectional Descriptive study. Setting: Microbiology laboratory, PGMI, Lahore. Period: January 2015 to December 2015. Material & Methods: 53 imipenem resistant NFGNB that were isolated from appropriate sampling of patients suffering from several infections were analyzed by using different standard microbiological techniques like microscopy, culture methods, biochemical reactions and antibiotic susceptibility using Kirby-Bauer method. MBL recognition was performed by imipenem-2MPA double disc synergy test (DDST). Results: This study shows the frequency of imipenem resistant non-fermenter Gram-negative bacilli isolated from various clinical wards. Maximum NFGNB were recovered from surgery/surgical allied 35.84% followed by ICU 28.3%, medicine /medicine allied 20.75%, pediatrics 9.4% and gynae/obs 5.6% respectively. MBL production was identified among different imipenem resistant non-fermenter Gram-negative bacilli isolates by DDST using 2-MPA. Out of total 53 imipenem resistant non-fermenter Gram Negative Bacilli 37 Pseudomonas aeruginosa 20(54.05%) were MBL positive. Out of 13 Acinetobacter baumannii and 2 Pseudomonas luteola, 11(84.61%) Acinetobacter baumannii and 1(50%) Pseudomonas luteola were positive for MBL production. None of the Acinetobacter junii indicated MBL production. Conclusion: Double disc synergy test is operational for detection of MBL producers among NFGNB. It can be established in our routine clinical microbiology laboratories, for the MBL recognition especially in imipenem resistant isolates as of its cost efficiency.

scholarly journals Phenotypic and Molecular Detection of IMP and SPM Metallo-Beta-Lactamases in Clinical Isolates of Carbapenem Resistant Pseudomonas aeruginosa

2020 ◽  
Author(s):  
Zahra Norouzi Bazgir ◽  
Mohammad Ahanjan ◽  
Hamid Reza Goli ◽  
Roya Ghasemian ◽  
Mohammad Bagher Hashemi-Soteh
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Susceptibility ◽  
Molecular Detection ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
Antibiotic Susceptibility Pattern ◽  
Beta Lactamases ◽  
Phenotypic Identification ◽  
Carbapenem Resistant ◽  
High Prevalence

Abstract Objectives: Metallo-beta-lactamases play a major role in the resistance of Pseudomonas aeruginosa to carbapenems. The aim of this study was the phenotypic and molecular detection of IMP and SPM carbapenemase genes in 100 carbapenem-resistant clinical isolates of P. aeruginosa. The isolates identified using standard microbiological tests, and their antibiotic susceptibility pattern determined by disk agar diffusion (Kirby Bauer) method. Phenotypic identification of Metallo-beta-lactamase-producing strains assessed by the combined disk test (CDT). Then, PCR was used to detect the presence of IMP and SPM genes.Results: The highest and lowest levels of antibiotic resistance were observed against gentamicin (40%) and piperacillin-tazobactam (13%), respectively. Besides, 40 isolates (40%) had the Multi-drug Resistant (MDR) phenotype, while 5 (12.5%) MDR isolates were resistant to all antibiotics tested. The results of the CDT showed that among 43 carbapenem non-susceptible clinical isolates of P. aeruginosa, 33 (76.74%) isolates were Metallo-beta-lactamase-producing strains. Also, the frequency of the IMP gene was determined to be 9%, while none of these isolates carried the SPM gene. Due to the high prevalence of carbapenem-resistant and MDR P. aeruginosa in this study, routine antibiotic susceptibility testing and phenotypic identification of carbapenemase production by this bacterium are necessary for proper selection of antibiotics.

scholarly journals Detection of Extended-Spectrum β-Lactamases in Clinical Isolates of Pseudomonas aeruginosa

2006 ◽  
Vol 50 (9) ◽  
pp. 2990-2995 ◽  
Author(s):  
Xiaofei Jiang ◽  
Zhe Zhang ◽  
Min Li ◽  
Danqiu Zhou ◽  
Feiyi Ruan ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Screening Methods ◽  
Extended Spectrum ◽  
Pcr Product ◽  
Amoxicillin Clavulanate ◽  
Synergy Test ◽  
Esbl Producers ◽  
Disk Test

ABSTRACT With the occurrence of extended-spectrum β-lactamases (ESBLs) in Pseudomonas aeruginosa being increasingly reported worldwide, there is a need for a reliable test to detect ESBLs in clinical isolates of P. aeruginosa. In our study, a total of 75 clinical isolates of P. aeruginosa were studied. Nitrocefin tests were performed to detect the β-lactamase enzyme; isoelectric focusing electrophoresis, PCR, and PCR product sequencing were designed to further characterize the contained ESBLs. Various ESBL-screening methods were designed to compare the reliabilities of detecting ESBLs in clinical isolates of P. aeruginosa whose β-lactamases were well characterized. Thirty-four of 36 multidrug-resistant P. aeruginosa clinical isolates were positive for ESBLs. bla VEB-3 was the most prevalent ESBL gene in P. aeruginosa in our study. Among the total of 34 isolates that were considered ESBL producers, 20 strains were positive using conventional combined disk tests and 10 strains were positive using a conventional double-disk synergy test (DDST) with amoxicillin-clavulanate, expanded-spectrum cephalosporins, aztreonam, and cefepime. Modifications of the combined disk test and DDST, which consisted of shorter distances between disks (20 mm instead of 30 mm) and the use of three different plates that contained cloxacillin (200 μg/ml) alone, Phe-Arg β-naphthylamide dihydrochloride (MC-207,110; 20 μg/ml) alone, and both cloxacillin (200 μg/ml) and MC-207,110 (20 μg/ml) increased the sensitivity of the tests to 78.8%, 91.18%, 85.29%, and 97.06%.

scholarly journals Molecular aspects of high-level resistance to sulbactam-cefoperazone in Klebsiella oxytoca clinical isolates.

1996 ◽  
Vol 40 (9) ◽  
pp. 1988-1994 ◽  
Author(s):  
K Kimura ◽  
Y Arakawa ◽  
S Ohsuka ◽  
H Ito ◽  
K Suzuki ◽  
...  
Keyword(s):  
Klebsiella Oxytoca ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Point Of View ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
Beta Lactamases ◽  
Molecular Aspects ◽  
High Level ◽  
Level Resistance

Nine Klebsiella oxytoca strains which demonstrated resistance to the combination of sulbactam and cefoperazone were isolated from geographically separate hospitals in Japan in 1995. Among them, K. oxytoca SB23 showed high-level resistance to sulbactam-cefoperazone (MIC > 128 micrograms/ml) and aztreonam (MIC, 128 micrograms/ml). The sulbactam-cefoperazone resistance was not transferred from strain SB23 to Escherichia coli CSH2 by conjugation, beta-Lactamase RbiA, produced by strain SB23, was purified, and the molecular mass was estimated to be 29 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Kinetic parameters for RbiA revealed that cefoperazone and aztreonam were hydrolyzed efficiently by this enzyme. Moreover, ceftazidime and imipenem were also hydrolyzed weakly by RbiA, although strain SB23 did not show any resistance to these agents. Clavulanate, sulbactam, and tazobactam failed to block the hydrolysis of cefoperazone by RbiA. The structural gene of RbiA (blaRBI) was cloned and sequenced, and the deduced amino acid sequence of RbiA demonstrated high-level similarities to those of the beta-lactamases found in K. oxytoca D488, E23004, and plasmid-mediated MEN-1, which have been classified into Bush functional group 2be. Although RbiA demonstrates high-level molecular similarity to the enzymes in group 2be, from an enzymological point of view, this enzyme might be differentiated from the enzymes in that group. Hybridization analysis revealed that beta-lactamase genes highly similar to blaRBI were generally encoded on the chromosome of the sulbactam-cefoperazone-resistant clinical isolates of K. oxytoca tested in the study, despite their different derivations. This observation suggests that sulbactam-cefoperazone-resistant A. oxytoca strains which produce RbiA-type beta-lactamases have been proliferating in many hospitals in Japan.

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

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

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

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