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

Author(s):  
Ashna Bhasin Poonam Loomba ◽  
Abha Sharma Bibhabati Mishra ◽  
Ashish Bajaj

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.

2010 ◽  
Vol 4 (04) ◽  
pp. 239-242 ◽  
Author(s):  
Supriya Upadhyay ◽  
Malay Ranjan Sen ◽  
Amitabha Bhattacharjee

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.


2020 ◽  
Vol 14 (3) ◽  
pp. 2027-2032
Author(s):  
Mita D. Wadekar ◽  
J.V. Sathish ◽  
C. Pooja ◽  
S. Jayashree

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.


Antibiotics ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1403
Author(s):  
Josman Dantas Palmeira ◽  
Marisa Haenni ◽  
Jean-Yves Madec ◽  
Helena Maria Neto Ferreira

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.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mojisola C. Hosu ◽  
Sandeep D. Vasaikar ◽  
Grace E. Okuthe ◽  
Teke Apalata

AbstractThe proliferation of extended spectrum beta-lactamase (ESBL) producing Pseudomonas aeruginosa represent a major public health threat. In this study, we evaluated the antimicrobial resistance patterns of P. aeruginosa strains and characterized the ESBLs and Metallo- β-lactamases (MBL) produced. Strains of P. aeruginosa cultured from patients who attended Nelson Mandela Academic Hospital and other clinics in the four district municipalities of the Eastern Cape between August 2017 and May 2019 were identified; antimicrobial susceptibility testing was carried out against thirteen clinically relevant antibiotics using the BioMérieux VITEK 2 and confirmed by Beckman autoSCAN-4 System. Real-time PCR was done using Roche Light Cycler 2.0 to detect the presence of ESBLs; blaSHV, blaTEM and blaCTX-M genes; and MBLs; blaIMP, blaVIM. Strains of P. aeruginosa demonstrated resistance to wide-ranging clinically relevant antibiotics including piperacillin (64.2%), followed by aztreonam (57.8%), cefepime (51.5%), ceftazidime (51.0%), piperacillin/tazobactam (50.5%), and imipenem (46.6%). A total of 75 (36.8%) multidrug-resistant (MDR) strains were observed of the total pool of isolates. The blaTEM, blaSHV and blaCTX-M was detected in 79.3%, 69.5% and 31.7% isolates (n = 82), respectively. The blaIMP was detected in 1.25% while no blaVIM was detected in any of the strains tested. The study showed a high rate of MDR P. aeruginosa in our setting. The vast majority of these resistant strains carried blaTEM and blaSHV genes. Continuous monitoring of antimicrobial resistance and strict compliance towards infection prevention and control practices are the best defence against spread of MDR P. aeruginosa.


2011 ◽  
Vol 2 (1) ◽  
pp. 8
Author(s):  
Ronak Bakhtiari ◽  
Jalil Fallah Mehrabadi ◽  
Hedroosha Molla Agamirzaei ◽  
Ailar Sabbaghi ◽  
Mohammad Mehdi Soltan Dallal

Resistance to b-lactam antibiotics by gramnegative bacteria, especially <em>Escherichia coli (E. coli)</em>, is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly <em>E. coli </em>is via the production of extended spectrum beta lactamase (ESBLs) enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk). Five hundred clinical samples were collected and 200 <em>E. coli </em>isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on <em>E. coli </em>isolates. A confirmation test (Combined Disk method) was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64%) <em>E. coli </em>isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8%) isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.


2015 ◽  
Vol 8 (1) ◽  
pp. 71-75 ◽  
Author(s):  
Olga I Chub ◽  
Aleksandr V Bilchenko ◽  
Igor Khalin

Background : Increased multidrug resistance of extended-spectrum beta-lactamases (ESBLs) compromises the efficacy of treatment of urinary tract infections. Objective : The objective of this study is to determine the prevalence of ESBL-producing uropathogens from hospitalized patients with chronic pyelonephritis and to identify the presence of genes involved in the resistance. Methods : A cross-sectional study of 105 patients with chronic pyelonephritis, treated in Kharkiv City Clinical Emergency Hospital, Ukraine was carried. Bacterial isolates were collected, antimicrobial susceptibility of isolates was determined by the Kirby Bauer disk diffusion method and screening for the presence of blaSHV, blaTEM, blaCTX-M ESBL genes was performed by polymerase chain reaction. Results : 84 (80%) patients had positive urine cultures. Eschеrichia coli wаs the most common microorganism isolated. Among them, 29 (25.2%) were found to be ESBL producers. Out of 53 E. coli isolates, 10 (18.9%), 4 (7.5%) and 6 (11.3%) were identified to carry bla(TEM), bla(SHV) and bla(CTX-M) beta-lactamase genes, respectively. The highest resistance was observed against ampicillin (75.9%), ciprofloxacin (48.3%), levofloxacin (41.4%) and gentamicin (41.4%). Beside this, only meropenem (96.6% susceptibility), nitroxolinum (86.2%) and fosfomycin (72.4%) exhibited a good enough activity against ESBLs-producing urinary strains. Conclusion : Isоlation and detеction of ESBL-prоducing strаins are еssential fоr the sеlection оf the mоst effеctive antibiоtic for the empiric trеatment.


1996 ◽  
Vol 40 (11) ◽  
pp. 2488-2493 ◽  
Author(s):  
P Mugnier ◽  
P Dubrous ◽  
I Casin ◽  
G Arlet ◽  
E Collatz

A clinical strain of Pseudomonas aeruginosa, PAe1100, was found to be resistant to all antipseudomonal beta-lactam antibiotics and to aminoglycosides, including gentamicin, amikacin, and isepamicin. PAe1100 produced two beta-lactamases, TEM-2 (pI 5.6) and a novel, TEM-derived extended-spectrum beta-lactamase called TEM-42 (pI 5.8), susceptible to inhibition by clavulanate, sulbactam, and tazobactam. Both enzymes, as well as the aminoglycoside resistance which resulted from AAC(3)-IIa and AAC(6')-I production, were encoded by an 18-kb nonconjugative plasmid, pLRM1, that could be transferred to Escherichia coli by transformation. The gene coding for TEM-42 had four mutations that led to as many amino acid substitutions with respect to TEM-2: Val for Ala at position 42 (Ala42), Ser for Gly238, Lys for Glu240, and Met for Thr265 (Ambler numbering). The double mutation Ser for Gly238 and Lys for Glu240, which has so far only been described in SHV-type but not TEM-type enzymes, conferred concomitant high-level resistance to cefotaxime and ceftazidime. The novel, TEM-derived extended-spectrum beta-lactamase appears to be the first of its class to be described in P. aeruginosa.


Antibiotics ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 67 ◽  
Author(s):  
Ahmed Zikri ◽  
Kamal El Masri

Infections, with multidrug-resistant Pseudomonas aeruginosa, are a major concern in the pediatric intensive care unit, especially in immunocompromised patients. Some of these strains are resistant to all beta-lactams, including carbapenems, leaving very limited treatment options remaining. These options include aminoglycosides and colistin, both of which have poor pharmacokinetic profiles with significant toxicities. Newer beta-lactam/beta-lactamase inhibitor combinations offer additional novel options to treat such infections, given their good pharmacokinetic profiles and activity against multi-drug resistant strains. Ceftolozane/tazobactam is a novel cephalosporin/beta-lactamase inhibitor combination approved in 2014. The drug demonstrates good activity against multidrug-resistant P. aeruginosa strains, including those resistant to all other antibiotics. Ceftolozane/tazobactam is currently approved in adult patients 18 years and older only. There are very limited data on its pharmacokinetic profile and clinical utility in the pediatric population. We report the use of ceftolozane/tazobactam to successfully treat pneumonia caused by multidrug-resistant P. aeruginosa in a pediatric patient with combined immunodeficiency syndrome.


2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Mohammad Sadegh Rezai ◽  
Ebrahim Salehifar ◽  
Alireza Rafiei ◽  
Taimour Langaee ◽  
Mohammadreza Rafati ◽  
...  

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.


2020 ◽  
Author(s):  
Carissa Duru ◽  
Grace M Olanipekun ◽  
Vivian Odili ◽  
Nicholas J Kocmich ◽  
Amy Rezac ◽  
...  

AbstractBackgroundBacteremia is a leading cause of death in developing countries but etiologic evaluation is infrequent and empiric antibiotics are not evidence-based. Very little is known about the types of extended-spectrum β-lactamases (ESBL) in pediatric bacteremia patients in Nigeria. We evaluated the patterns of ESBL resistance in children enrolled into surveillance for community acquired bacteremic syndromes across health facilities in Central and Northwestern Nigeria.MethodBlood culture from suspected cases of sepsis from children age less than 5 years were processed using automated Bactec® incubator System from Sept 2008-Dec 2016. Enterobacteriacea were identified to the species level using Analytical Profile Index (API20E®) identification strip and antibiotic susceptibility profile was determined by the disc diffusion method. The multidrug resistant strains were then screened and confirmed for extended spectrum beta lactamase (ESBL) production by the combination disc method as recommended by Clinical and Laboratory Standard Institute (CLSI). Real time PCR was used to elucidate the genes responsible for ESBL production characterize the resistance genesResultOf 21,000 children screened from Sept 2008-Dec 2016, 2,625(12.5%) were culture-positive. A total of 413 Enterobacteriaceae available for analysis were screened for ESBL. ESBL production was detected in 160/413(38.7%), comprising Klebsiella pneumoniae 105/160(65.6%), Enterobacter cloacae 21/160(13.1%), Escherichia coli 22/160(13.8%), Serratia species 4/160(2.5%), Pantoea species 7/160(4.4%) and Citrobacter species 1/160(0.6%). Of the 160 ESBL-producing isolates, high resistance rates were observed among ESBL-positive isolates for Ceftriaxone (92.3%), Aztreonam (96.8%), Cefpodoxime (96.25%), Cefotaxime (98.75%) and sulphamethoxazole-trimethoprim (90%), while 87.5 %, 90.63%, and 91.87% of the isolates were susceptible to Imipenem, Amikacin and Meropenem respectively. Frequently detected resistance genes were blaTEM 83.75%) (134/160), and, blaCTX-M 83.12% (133/160) followed by blaSHV genes 66.25% (106/160). Co-existence of blaCTX-M, blaTEM and blaSHV was seen in 94/160 (58.8%), blaCTX-M and blaTEM in 118/160 (73.8%), blaTEM and blaSHV in 97/160 (60.6%) and blaCTX-M and blaSHV in 100/160 (62.5%) of isolates tested.ConclusionOur results indicate a high prevalence of ESBL resistance to commonly used antibiotics in Enterobacteriaceae isolates from bloodstream infections in children in this study. Careful choice of antibiotic treatment options and further studies to evaluate transmission dynamics of resistance genes could help in the reduction of ESBL resistance in these settings.


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