scholarly journals Vancomycin and high-level aminoglycoside resistance in Enterococcus species

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Seyda Ozarslan Kurtgoz ◽  
Burcin Ozer ◽  
Melek Inci ◽  
Nizami Duran ◽  
Erkan Yula
Keyword(s):  
Vancomycin Resistance ◽  
Automated System ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Aminoglycoside Resistance ◽  
Disk Diffusion Method ◽  
Gradient Diffusion ◽  
Gentamicin Resistance ◽  
Polymerase Chain ◽  
High Level

The aim of the study was to investigate vancomycin and high-level aminoglycoside resistance (HLAR) in <em>Enterococcus</em> species by phenotypic and genotypic methods. A hundred <em>Enterococcus</em> strains were included in the study. Antimicrobial susceptibilities of strains were investigated by automated system, betalactamase production was investigated by nitrocefin disks, vancomycin resistance and HLAR were investigated by gradient diffusion method (GDM) and disk diffusion method, respectively. For detection of vancomycin and high-level gentamicin resistance (HLGR) genes, polymerase chain reaction was used. Teicoplanin linezolid, vancomycin, ampicillin, penicillin are the most susceptible antibiotics and strains were detected not to produce beta lactamase. Vancomycin resistance was detected in ten isolates by automated system and in only five isolates by GDM. Five isolates carrying <em>VanA</em> gene were determined. The ratio of HLGR and high-level streptomycin resistance was found 40 and 63% respectively. <em>aac (6’)-1eaph (2’’)-1a</em> gene was detected in 58% of strains. <em>E. faecium</em> strains were found more resistant to the antibiotics than the other species. Beta lactamase was detected in none of strains. The automated system detected vancomycin resistance in more strains than GDM. Therefore it is concluded that strains, which were detected to be resistant to vancomycin, should be confirmed by GDM. The ratio of <em>VanA</em> gene in strains is consistent with other studies. The HLAR ratio was found in about half of strains. The ratio of<em> aac(6’)-1e-aph(2’’)-1a</em> gene, which is the most reported gene in our country and other countries and one of the HLGR genes investigated in our study, was detected 58%.

scholarly journals Prevalence of Transferable OXA-1 β-Lactamase Associated with Carbapenem-Resistant Klebsiella pneumoniae Isolates in Iraq

2021 ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Klebsiella Pneumoniae ◽  
Diffusion Method ◽  
Antibiotics Resistance ◽  
Disk Diffusion Method ◽  
Antibiotic Resistance Profile ◽  
Carbapenem Resistant ◽  
Stool Samples ◽  
Polymerase Chain ◽  
High Level ◽  
Conjugation Experiment

This study was designed to explore the incidence of blaOXA-1 amongst Klebsiella pneumoniae isolates with resistant to carbapenem. Between December 2014 and April 2015, one hundred samples were taken from two hospitals: Babylon Teaching Hospital for Maternity and Pediatric / Babylon Province (clinical, umbilical infections, n= 40; environmental, n=20) and Karbala Hospital for Pediatric / Karbala Province (40 stool samples). All patients were hospitalized or attended these hospitals, all under 1 year of age. Seventeenth (17%) isolates were identified as Klebsiella pneumoniae. The antibiotic resistance profile of isolates was tested using disk diffusion method. High-level of resistance was recorded with ampicillin (94.1%) and piperacillin (88.2%) antibiotics. Resistance to carbapenem was reported in two K.pneumoniae isolates, these were investigated for the existence of OXA-1b-lactamase using Polymerase Chain Reaction (PCR) technique. Two (100%) isolates gave positive result. Transference of this gene was studied by conjugation experiment. The blaOXA-1 gene conjugated successfully in 1 (50%) isolate only.

scholarly journals Analysis of Aminoglycoside Modifying Enzyme Genes Responsible for High-Level Aminoglycoside Resistance among Enterococcal Isolates

2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Vishal Shete ◽  
Naveen Grover ◽  
Mahadevan Kumar
Keyword(s):  
Active Agent ◽  
Bactericidal Effect ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Aminoglycoside Resistance ◽  
Enzyme Gene ◽  
A Cell ◽  
Polymerase Chain ◽  
High Level ◽  
Level Resistance

Enzymatic modification results in high-level resistance to aminoglycoside (HLAR), which eliminates the synergistic bactericidal effect of combined exposure to a cell wall-active agent and an aminoglycoside. So aim of the study was to determine prevalence of HLAR enterococcal isolate and to study distribution of aminoglycoside modifying enzyme genes in them. A total of 100 nonrepeat isolates of enterococci from various clinical samples were analyzed. As per Clinical and Laboratory Standards Institute guidelines enterococci were screened for HLAR by Kirby-Bauer disc diffusion method. Minimum inhibitory concentration of all isolates for gentamicin and streptomycin was determined by E-test. Multiplex polymerase chain reaction (PCR) was carried out for HLAR enterococcal isolates to identify aminoglycoside modifying enzymes genes responsible for resistance. 60% isolates were found to be high-level gentamicin resistant (HLGR) whereas 45% isolates were found to be high-level streptomycin resistant (HLSR). By multiplex PCR 80% HLGR isolates carried bifunctional aminoglycoside modifying enzyme geneaac(6′)-Ie-aph(2′′)-Iawhereas 18 out of 45 high-level streptomycin resistant, that is, 40%, isolates carriedaph(3′)-IIIa.However,aph(2′′)-Ib, aph(2′′)-Ic, aph(2′′)-Id,andant(4′)-Iagenes which encode other aminoglycosides modifying enzymes were not detected. Bifunctional aminoglycoside modifying enzyme geneaac(6′)-Ie-aph(2′′)-Iais the predominant gene responsible for HLAR.

scholarly journals Increasing relevance of Gram-positive cocci in urinary tract infections: a 10-year analysis of their prevalence and resistance trends

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Márió Gajdács ◽  
Marianna Ábrók ◽  
Andrea Lázár ◽  
Katalin Burián
Keyword(s):  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Diffusion Method ◽  
Aminoglycoside Resistance ◽  
Gram Positive ◽  
Disk Diffusion Method ◽  
Pronounced Increase ◽  
High Level ◽  
Tract Infections ◽  
Gram Positive Cocci

Abstract Urinary tract infections (UTIs) are the third most common types of infection in human medicine worldwide. There is increasing appreciation for the pathogenic role of Gram-positive cocci (GPC) in UTIs, as they have a plethora of virulence factors, maintaining their pathogenicity and high affinity for the epithelial cells of the urinary tract. The study was carried out using microbiological data collected corresponding to the period between 2008 and 2017. Antimicrobial susceptibility testing was performed using the disk diffusion method and E-tests. The age range of patients affected from the outpatient and inpatient groups differed significantly (43 [range 0.7–99] vs. 68 [range 0.4–99] years; p = 0.008). 3962 GPCs were obtained from inpatient and 4358 from outpatient samples, corresponding to 20.5 ± 2.8% (range 17.5–26.8%) and 20.6 ± 2.6% (range 17.8–26.0%) of all positive urine samples (p > 0.05); in both groups, Enterococcus spp. were the most prevalent (outpatients: 79.6%; inpatients: 88.5%). High-level aminoglycoside resistance in enterococci was noted in 31.0–46.6% of cases. A pronounced increase in the number of MRSA was seen in the second half of the study period (0.6–1.9% vs. 9.8–11.6%; p = 0.038). The ratio of VRE isolates was 0.16%, no VISA/VRSA isolates were detected.

scholarly journals Phenotypic detection of beta-lactamases production in enterobacteriaceae

2014 ◽  
Vol 142 (7-8) ◽  
pp. 457-463 ◽  
Author(s):  
Ivana Cirkovic ◽  
Ljiljana Pavlovic ◽  
Neda Konstantinovic ◽  
Katarina Kostic ◽  
Snezana Jovanovic ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Automated System ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Beta Lactamases ◽  
Beta Lactam Antibiotics ◽  
Clinical Center ◽  
Different Types

Introduction. Beta-lactam antibiotics are the most commonly used antibacterial drugs. However, many bacteria have developed resistance to these antibiotics, and the most common form of resistance is the production of beta-lactamase enzymes. Many members of the Enterobacteriaceae family produce different types of these enzymes. Objective. The aim of this study was to perform phenotypic detection of production and identification of beta-lactamase type in Enterobacteriaceae isolated from different clinical specimens from patients hospitalized in the Clinical Center of Serbia. Methods. The strains of Enterobacteriaceae were collected between November 2011 and January 2012 in the laboratory of the Clinical Center of Serbia. The isolates were identified according to the standard microbiology procedures and confirmed by the Vitek2 automated system. Antimicrobial susceptibility testing was performed by the disk diffusion method, and the phenotypic detection of production and identification of betalactamases was performed according to previously described methodologies. Results. In this study, a total of 172 Enterobacteriaceae strains were isolated. Further testing was performed on 54/145 (37.2%) strains showing decreased susceptibility to beta-lactam antibiotics: 13/85 (15.3%) Escherichia coli, 31/46 (67.4%) Klebsiella pneumoniae and 10/14 (71.4%) Proteus mirabilis. Among them, 40/145 (27.6%) strains produced extended spectrum betalactamases (ESBLs), 9/145 (6.2%) - AmpC, 1/145 (0.7%) - K1 beta-lactamase and 4/145 (2.8%) - carbapenemases. Carbapenemases were predominantly detected in K. pneumoniae (75%). Conclusion. Enterobacteriaceae produce different types of betalactamases, and the most common type in our study was ESBLs. Production of carbapenemases detected in Enterobacteriaceae is also an associated problem.

Virulence characterization of Klebsiella pneumoniae and its relation with ESBL and AmpC beta-lactamase associated resistance

2020 ◽  
Author(s):  
Elghar Soltani ◽  
Alka Hasani ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Tahereh Pirzadeh ◽  
Mahin Ahangar Oskouee ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Virulence Genes ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Disk Diffusion Method ◽  
Extended Spectrum Beta Lactamase ◽  
Virulence Trait ◽  
Polymerase Chain

Background and Objectives: Trend analysis reveals that Klebsiella pneumoniae has witnessed a steep enhancement in the antibiotic resistance and virulence over the last few decades. The present investigation aimed at a comprehensive approach investigating antibiotic susceptibility including, extended spectrum beta-lactamase (ESBL) and AmpC β-lactamase (AmpC) resistance and the prevalence of virulence genes among the K. pneumoniae isolates. Materials and Methods: Sixty-one K. pneumoniae isolates were obtained from various clinical infections. Antimicrobial susceptibility was performed by disk diffusion method. The Mast® D68C test detected the presence of ESBLs and AmpCs phenotypically, and later presence of ESBL and AmpC genes was observed by polymerase chain reaction (PCR). Multi- plex-PCR was performed to investigate various virulence genes. CMY-2   Results: Amongst 61 K. pneumoniae isolates, 59% were observed as ESBL and 14.7% as AmpC producers. All ESBL   CTX-M-15   producers were positive for bla   CTX-M-15   , while bla   CTX-M-14   was observed in 54.1% isolates. The frequency of AmpC genes was   CMY-2   as follows: bla   CMY-2   (60.7%) and bla   DHA-1   (34.4%). The most frequent virulence genes were those encoding enterobactin and   DHA-1   lipopolysaccharide. Presence of mrkD was associated with bla   CMY-2   DHA-1   gene, while bla   significantly (p≤0.05) correlated   DHA-1   with the presence of iutA and rmpA virulence genes. bla   positive isolates had urine as a significant source, while bla   positive isolates were mainly collected from wound exudates (p≤0.05). Conclusion: Our results highlight that ESBL and AmpC production along with a plethora of virulence trait on K. pneumoni- ae should be adequately considered to assess its pathogenesis and antibiotic resistance.  

Prevalence of SHV-extended spectrum β-lactamase producing carbapenem –resistantKlebsiellapneumoniae among patients with lower respiratory tractinfections in Babylon Province-Iraq.

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Resistance Rate ◽  
Combination Method ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Extended Spectrum ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections ◽  
Positive Results

This study was carried out to screen the prevalence of Klebsiella pneumoniae isolated from patients with lower respiratory tract infections in Babylon province.From December,2015 to the end of March,2016,a total of 100 sputum samples were collected from patients visited or hospitalized Merjan Teaching Hospital and Al- Hashimya General Hospital. Fifteenth (65%) isolates were identified as Klebsiellapneumoniae. All bacterial isolates were evaluated for extended spectrum β-lactamase (ESBL) production phenotypically using disk combination method. Eleven (73.3%) isolates were detected as ESBL-producers. Kirby-Bauer disk diffusion method was employed to determine resistance profile of ESBLs-positive isolates. Higher rates of resistance were observed for ampicillin and piperacillin antibiotics with (81.8%) and (72.7%) resistance rate, respectively, while the lowest rate was noticed for imipenem antibiotic (14.28%). Carbapenem-resistant isolates were investigated for blaSHV gene by Polymerase Chain Reaction (PCR) method, 2 (100%) isolates gave positive results.

scholarly journals Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

2011 ◽  
Vol 2 (1) ◽  
pp. 8
Author(s):  
Ronak Bakhtiari ◽  
Jalil Fallah Mehrabadi ◽  
Hedroosha Molla Agamirzaei ◽  
Ailar Sabbaghi ◽  
Mohammad Mehdi Soltan Dallal
Keyword(s):  
Escherichia Coli ◽  
Disk Diffusion ◽  
Confirmatory Test ◽  
Diffusion Method ◽  
Multi Drug Resistance ◽  
Beta Lactamase ◽  
Disk Diffusion Method ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum

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.

scholarly journals Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Raw Milk ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

scholarly journals The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002

2004 ◽  
Vol 61 (4) ◽  
pp. 391-397
Author(s):  
Veljko Mirovic ◽  
Branka Tomanovic ◽  
Sonja Konstantinovic
Keyword(s):  
Clinical Laboratory ◽  
Blood Cultures ◽  
Diffusion Method ◽  
National Committee ◽  
Disk Diffusion Method ◽  
Beta Lactamases ◽  
Resistance To Antibiotics ◽  
Extended Spectrum Beta Lactamases ◽  
High Level

The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002. The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures. The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%), during the follow-up period. None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2-72.2%). In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA fenotype). Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1%) during the follow-up period. Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001). Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates. Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time. Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period.

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

2010 ◽  
Vol 4 (04) ◽  
pp. 239-242 ◽  
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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