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Sample records for beta-lactamases

  1. Metalo-beta-lactamases Metallo-beta-lactamases

    Directory of Open Access Journals (Sweden)

    Rodrigo Elisandro Mendes

    2006-04-01

    Full Text Available Nos últimos anos tem sido observada maior incidência de bacilos Gram-negativos resistentes a cefalosporinas de espectro ampliado no ambiente hospitalar, ocasionando, assim, maior uso de betalactâmicos mais potentes, como os carbapenens. A utilização de carbapenens exerce maior pressão seletiva sobre a microbiota hospitalar, o que pode ocasionar aumento da resistência a esses agentes. Entre os mecanismos de resistência a carbapenens mais comumente identificados estão a produção de betalactamases, como, por exemplo, as pertencentes à classe D de Ambler e as que pertencem à classe B de Ambler, ou metalo-beta-lactamases (MbetaL. Essas últimas hidrolisam todos betalactâmicos comercialmente disponíveis, sendo a única exceção o monobactam aztreonam. Desde o início da década de 1990, novos genes que codificam MbetaLs têm sido descritos em microrganismos clinicamente importantes, como Pseudomonas spp., Acinetobacter spp. e membros da família Enterobacteriaceae. O encontro desses microrganismos não-sensíveis a carbapenens pode ser submetido a metodologias fenotípicas para detecção da produção de MbetaL com o intuito de auxiliar a Comissão de Controle de Infecção Hospitalar (CCIH e prevenir a disseminação desses determinantes de resistência, uma vez que genes que codificam MbetaLs estão contidos em estruturas genéticas que propiciam sua mobilidade de forma muito efetiva, sendo então facilmente disseminados.Increase isolation of Gram-negative bacilli resistant to broad-spectrum cephalosporin has been observed during the last few years, thus determining the use of more potent beta-lactams, such as carbapenems. The use of these antimicrobial agents may lead to the emergence of carbapenem resistant Gram-negative bacilli in the nosocomial environment. Carbapenem resistance may be due to the production of Ambler class D beta-lactamase or Ambler class B beta-lactamase, also called metallo-beta-lactamase (MbetaL. Apart from

  2. Antibodies against chromosomal beta-lactamase

    DEFF Research Database (Denmark)

    Giwercman, B; Rasmussen, J W; Ciofu, Oana; Clemmentsen, I; Schumacher, H; Høiby, N

    1994-01-01

    A murine monoclonal anti-chromosomal beta-lactamase antibody was developed and an immunoblotting technique was used to study the presence of serum and sputum antibodies against Pseudomonas aeruginosa chromosomal group 1 beta-lactamase in patients with cystic fibrosis (CF). The serum antibody...... response was studied with serum samples collected in 1992 from 56 CF patients in a cross-sectional study and with serum samples from 18 CF patients in a longitudinal study. Anti-beta-lactamase immunoglobulin G antibodies were present in all of the serum samples from the patients with chronic...... bronchopulmonary P. aeruginosa infection (CF + P) but in none of the CF patients with no or intermittent P. aeruginosa infection. Anti-beta-lactamase antibodies were present in serum from CF + P patients after six antipseudomonal courses (median) and correlated with infection with a beta-lactam-resistant strain of...

  3. Induction of beta-lactamase production in Pseudomonas aeruginosa biofilm

    DEFF Research Database (Denmark)

    Giwercman, B; Jensen, E T; Høiby, N;

    1991-01-01

    Imipenem induced high levels of beta-lactamase production in Pseudomonas aeruginosa biofilms. Piperacillin also induced beta-lactamase production in these biofilms but to a lesser degree. The combination of beta-lactamase production with other protective properties of the biofilm mode of growth...

  4. Beta-Lactamase Inhibitory Potential and Antibacterial Potentiation of Certain Medicinal Plants and Extracts against Extended Spectrum Beta-Lactamase Producers

    OpenAIRE

    Selvanayagam M; Shakti Singh Solanki

    2013-01-01

    Beta-lactamase inhibitory potential of fifteen plants was checked by beta-lactamase enzyme inhibition assay wherein inhibition of commercial beta-lactamase by their extracts was determined using a chromogenic substrate. These plant extracts showed considerable beta-lactamase inhibition. All these plant extracts were further checked for their inhibition of Extended Spectrum Beta-lactamase in the screening assay. Eight of the plant extracts showed potentiation of cephalosporins against the Exte...

  5. Characterization of beta-lactamases in situ on polyacrylamide gels.

    OpenAIRE

    Sanders, C C; Sanders, W E; Moland, E S

    1986-01-01

    An inhibitor-based characterization system which allowed the identification of beta-lactamases after isoelectric focusing on polyacrylamide gels was developed. This system, using potassium clavulanate and oxacillin, distinguished type I chromosomally mediated enzymes from other beta-lactamases of gram-negative bacteria.

  6. [beta]-Lactamases in the Biochemistry and Molecular Biology Laboratory

    Science.gov (United States)

    Amador, Paula; Prudencio, Cristina; Vieira, Monica; Ferraz, Ricardo; Fonte, Rosalia; Silva, Nuno; Coelho, Pedro; Fernandes, Ruben

    2009-01-01

    [beta]-lactamases are hydrolytic enzymes that inactivate the [beta]-lactam ring of antibiotics such as penicillins and cephalosporins. The major diversity of studies carried out until now have mainly focused on the characterization of [beta]-lactamases recovered among clinical isolates of Gram-positive staphylococci and Gram-negative…

  7. Phenotypic detection of beta-lactamases production in enterobacteriaceae

    OpenAIRE

    Ćirković Ivana; Pavlović Ljiljana; Konstantinović Neda; Kostić Katarina; Jovanović Snežana; Đukić Slobodanka

    2014-01-01

    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 Ente...

  8. [Transfer of plasmid beta-lactamases in enterobacteria].

    Science.gov (United States)

    Umaran, A; Garaizar, J; Gallego, L; Colom, K; Cisterna, R

    1989-04-01

    The aim of the present study was to determine which types of beta-lactamases codified by plasmids are transferred by conjugation from several species of enterobacteria. To this end, 352 strains of ampicillin-resistant enterobacteria from clinical samples from the Hospital Civil of Bilbao were evaluated. Their beta-lactamase activity and their capacity to transfer this capacity by conjugation were evaluated. The several types of plasmidic beta-lactamases in the strains that conjugated and in their respective transconjugants were characterized by analytic isoelectric approach, and also the sensitivity of these stains to 20 beta-lactamic antibiotics and the size of their plasmids. Twenty different types were detected, with a clear predominance of TEM 1. Type TEM 2 was found in 19% of the strains which conjugated, and much less commonly the types SHV 1, HMS 1 and a beta-lactamase of an approximate pl of 4.9 were found. The transfer of these beta-lactamases is mediated by a great variety of plasmids and is associated with variable levels of resistance to penicillins and unstable cephalosporins. The presence of betalactamases with activity on the more stable cephalosporins has not been detected. PMID:2490696

  9. Purification of Staphylococcus aureus beta-lactamases by using sequential cation-exchange and affinity chromatography.

    OpenAIRE

    Kernodle, D S; Zygmunt, D J; McGraw, P A; Chipley, J R

    1990-01-01

    Boronic acids are active-site inhibitors of serine beta-lactamases, and a phenylboronic acid-agarose affinity column has been used to purify beta-lactamase from crude cell extracts of several bacterial species. We applied phenylboronic acid-agarose chromatography to the purification of Staphylococcus aureus beta-lactamase. Two factors interfered with the success of the previously described single-step chromatographic protocol. First, staphylococcal beta-lactamase exhibited non-active-site-med...

  10. Comparative evaluation of a new beta-lactamase inhibitor, YTR 830, combined with different beta-lactam antibiotics against bacteria harboring known beta-lactamases.

    OpenAIRE

    Gutmann, L; Kitzis, M D; Yamabe, S; Acar, J F

    1986-01-01

    YTR 830, a new beta-lactamase inhibitor, combined with amoxicillin or carbenicillin, showed a synergistic effect similar to that observed with clavulanic acid, and generally better than that with sulbactam, against strains harboring chromosome-encoded penicillinases and broad-spectrum beta-lactamases or plasmid-determined beta-lactamases. With ampicillin, YTR 830 showed the best synergistic activity of the inhibitors against Proteus morganii, Citrobacter freundii, and Enterobacter cloacae and...

  11. Beta-lactamases in Enterobacteriaceae infections in children.

    Science.gov (United States)

    Moxon, Christopher Alan; Paulus, Stéphane

    2016-07-01

    Multi-drug resistance in Gram negative bacteria, particularly in Enterobacteriaceae, is a major clinical and public health challenge. The main mechanism of resistance in Enterobacteriaceae is linked to the production of beta-lactamase hydrolysing enzymes such as extended spectrum beta-lactamases (ESBL), AmpC beta-lactamases and carbapenemases (Carbapenemase Producing Enterobacteriaceae (CPE)). ESBL and CPE resistance genes are located on plasmids, which can be transmitted between Enterobacteriaceae, facilitating their spread in hospitals and communities. These plasmids usually harbour multiple additional co-resistance genes, including to trimethoprim-sulfamethoxazole, aminoglycosides, and fluoroquinolones, making these infections challenging to treat. Asymptomatic carriage in healthy children as well as community acquired infections are increasingly reported, particularly with ESBL. Therapeutic options are limited and previously little used antimicrobials such as fosfomycin and colistin have been re-introduced in clinical practice. Paediatric experience with these agents is limited hence there is a need to further examine their clinical efficacy, dosage and toxicity in children. Antimicrobial stewardship along with strict infection prevention and control practices need to be adopted widely in order to preserve currently available antimicrobials. The future development of novel agents effective against beta-lactamases producers and their applicability in children is urgently needed to address the challenge of multi-resistant Gram negative infections. PMID:27180312

  12. Docking and scoring of metallo-beta-lactamases inhibitors

    DEFF Research Database (Denmark)

    Olsen, Lars; Pettersson, Ingrid; Hemmingsen, Lars; Adolph, Hans-Werner; Jørgensen, Flemming Steen

    2004-01-01

    The performance of the AutoDock, GOLD and FlexX docking programs was evaluated for docking of dicarboxylic acid inhibitors into metallo-beta-lactamases (MBLs). GOLD provided the best overall performance, with RMSDs between experimental and docked structures of 1.8-2.6 A and a good correlation bet...

  13. Detection of resistance due to inducible beta-lactamase in Enterobacter aerogenes and Enterobacter cloacae.

    OpenAIRE

    Huber, T W; Thomas, J. S

    1994-01-01

    Thirty-six of 36 strains of Enterobacter cloacae and E. aerogenes with inducible beta-lactamase developed resistance when cefoxitin (inducer) was added to cefuroxime disks. Constitutive beta-lactamase producers (n = 23) were all resistant to cefuroxime. Cefuroxime resistance correlated with the amount of induced or constitutive beta-lactamase. Cefuroxime was a better indicator of induced resistance than cefamandole, cefazolin, cephalothin, ceftriaxone, cefotaxime, ticarcillin with or without ...

  14. Coexistence of Extended Spectrum Beta-Lactamases, AmpC Beta-Lactamases and Metallo-Beta-Lactamases in Acinetobacter baumannii from burns patients: a report from a tertiary care centre of India.

    Science.gov (United States)

    Gupta, V; Garg, R; Garg, S; Chander, J; Attri, A K

    2013-12-31

    Multidrug-resistant Acinetobacter baumanii is a major pathogen encountered in pyogenic infections, especially from burns patients in hospital settings. Often there is also coexistence of multiple beta-lactamase enzymes responsible for beta-lactam resistance in a single isolate, which further complicates treatment options. We conducted a study on burn wound pus samples obtained from the burns unit of our hospital. Phenotypic tests were used to determine the Extended Spectrum Beta-Lactamase, AmpC Beta-Lactamase and Metallo-Beta-Lactamase producing status of the isolates. Almost half of the samples from the burn wounds yielded Acinetobacter baumanii as the predominant pathogen (54.05%). Coexistence of the three resistance mechanisms was seen in 25 of the 100 (25%) isolates of Acinetobacter baumanii. This study emphasizes the need for the detection of isolates that produce these enzymes to avoid therapeutic failures and nosocomial outbreaks. PMID:24799848

  15. Coexistence of Extended Spectrum Beta-Lactamases, AmpC Beta-Lactamases and Metallo-Beta-Lactamases in Acinetobacter baumannii from burns patients: a report from a tertiary care centre of India

    OpenAIRE

    Gupta, V.; Garg, R.; Garg, S.; J Chander; Attri, A.K.

    2013-01-01

    Multidrug-resistant Acinetobacter baumanii is a major pathogen encountered in pyogenic infections, especially from burns patients in hospital settings. Often there is also coexistence of multiple beta-lactamase enzymes responsible for beta-lactam resistance in a single isolate, which further complicates treatment options. We conducted a study on burn wound pus samples obtained from the burns unit of our hospital. Phenotypic tests were used to determine the Extended Spectrum Beta-Lactamase, Am...

  16. Characterization of a new beta-lactamase gene from isolates of Vibrio spp. in Korea.

    Science.gov (United States)

    Jun, Lyu Jin; Kim, Jae Hoon; Jin, Ji Woong; Jeong, Hyun Do

    2012-04-01

    PCR was performed to analyze the beta-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known beta-lactamase genes. This prompted us to screen new beta-lactamase genes. A novel beta-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 beta-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other beta-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A beta-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 beta-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various beta-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 beta-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 beta- lactamase gene, led to the assumption that the location of this new beta-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 beta-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of

  17. Nanomolar Inhibitors of AmpC [beta]-Lactamase

    Energy Technology Data Exchange (ETDEWEB)

    Morandi, Federica; Caselli, Emilia; Morandi, Stefania; Focia, Pamela J.; Blazquez, Jesus; Shoichet, Brian K.; Prati, Fabio (Degali); (NIH); (NWU); (UCSF)

    2010-03-08

    {beta}-lactamases are the most widespread resistance mechanism to {beta}-lactam antibiotics, such as the penicillins and the cephalosporins. In an effort to combat these enzymes, a combination of stereoselective organic synthesis, enzymology, microbiology, and X-ray crystallography was used to design and evaluate new carboxyphenyl-glycylboronic acid transition-state analogue inhibitors of the class C {beta}-lactamase AmpC. The new compounds improve inhibition by over 2 orders of magnitude compared to analogous glycylboronic acids, with K{sub i} values as low as 1 nM. On the basis of the differential binding of different analogues, the introduced carboxylate alone contributes about 2.1 kcal/mol in affinity. This carboxylate corresponds to the ubiquitous C3(4)' carboxylate of {beta}-lactams, and this energy represents the first thermodynamic measurement of the importance of this group in molecular recognition by class C {beta}-lactamases. The structures of AmpC in complex with two of these inhibitors were determined by X-ray crystallography at 1.72 and 1.83 {angstrom} resolution. These structures suggest a structural basis for the high affinity of the new compounds and provide templates for further design. The highest affinity inhibitor was 5 orders of magnitude more selective for AmpC than for characteristic serine proteases, such as chymotrypsin. This inhibitor reversed the resistance of clinical pathogens to the third generation cephalosporin ceftazidime; it may serve as a lead compound for drug discovery to combat bacterial resistance to {beta}-lactam antibiotics.

  18. Phenotypic detection of beta-lactamases production in enterobacteriaceae

    Directory of Open Access Journals (Sweden)

    Ćirković Ivana

    2014-01-01

    Full Text Available 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. [Projekat Ministarstva nauke Republike Srbije, br. ON 175039

  19. New leads of metallo-beta-lactamase inhibitors from structure-based pharmacophore design

    DEFF Research Database (Denmark)

    Olsen, Lars; Jost, Sandra; Adolph, Hans-Werner;

    2006-01-01

    We have applied pharmacophore generation, database searching, docking methodologies, and experimental enzyme kinetics to discover new structures for design of di-zinc metallo-beta-lactamase inhibitors. Based on crystal structures of class B1 metallo-beta-lactamases with a succinic acid and a merc...

  20. Extended-spectrum plasmid-mediated beta-lactamases.

    Science.gov (United States)

    Sirot, D

    1995-07-01

    Extended-spectrum beta-lactamases (ESBLs) are mutant enzymes which derive from TEM or SHV (class A) enzymes. They confer variable levels of resistance to cefotaxime, ceftazidime and other broad-spectrum cephalosporins and to monobactams such as aztreonam but have no detectable activity against cephamycins and carbapenems. Recently, new plasmid-mediated ESBLs, not derived from TEM or SHV enzymes but related to cephalosporinases of Enterobacteriaceae (class C enzymes), that confer resistance to all cephalosporins including cephamycins, have been reported. However, to date there have been no reported outbreaks due to strains producing transferable cephalosporinases. Klebsiella pneumoniae is the species in which the ESBL enzymes have been most commonly reported around the world. Most of the clinical isolates that produce TEM- or SHV-derived ESBL, come from hospitalised patients and have frequently caused nosocomial outbreaks. Care should be taken in the selection of a beta-lactam for the treatment of infections because the presence of an ESBL does not prevent other mechanisms of resistance, such as decreased permeability, from emerging. Broad-spectrum cephalosporins including cefepime and cefpirome are hydrolysed by ESBL. However, low level resistance to cefotaxime, ceftriaxone, cefepime and aztreonam does occur in some strains producing certain TEM-derived ESBL. It remains to be seen, therefore, whether such isolates are clinically susceptible to these drugs. The combination of a third-generation cephalosporin and a beta-lactamase inhibitor such as sulbactam could be of interest against some strains producing certain ESBLs. Among the 7-alpha-methoxy cephalosporins, cefotetan and latamoxef are the most active. However, cephamycins should be used with caution to treat infections caused by ESBL-producing K. pneumoniae because of the relative ease with which clinical strains decrease the expression of outer membrane proteins. The most active beta-lactams are the

  1. Emergence of clinical isolates of Escherichia coli producing TEM-1 derivatives or an OXA-1 beta-lactamase conferring resistance to beta-lactamase inhibitors.

    OpenAIRE

    Zhou, X. Y.; Bordon, F; Sirot, D; Kitzis, M D; Gutmann, L

    1994-01-01

    Sixteen Escherichia coli clinical isolates which were resistant to ampicillin and amoxicillin-clavulanate but susceptible to cephalothin were studied. Eight strains showed the presence of a beta-lactamase which comigrates with reference OXA-1 enzyme. The eight other strains produced different TEM-1 derivatives which had in common a higher Km for penicillins and a higher 50% inhibitory concentration for the beta-lactamase inhibitors. By oligotyping and sequencing of PCR products, it was shown ...

  2. Bactericidal interactions of a beta-lactam and beta-lactamase inhibitors in experimental Pseudomonas aeruginosa endocarditis caused by a constitutive overproducer of type Id beta-lactamase.

    OpenAIRE

    Bayer, A S; Selecky, M; Babel, K; Hirano, L; Yih, J; Parr, T R

    1987-01-01

    We investigated the in vitro and in vivo effects of a combination of a beta-lactam (ceftazidime) and a beta-lactamase inhibitor (dicloxacillin) to synergistically kill a ceftazidime-resistant variant, Pseudomonas aeruginosa PA-48, which overproduces type Id cephalosporinase constitutively. In vitro, dicloxacillin plus ceftazidime exerted bactericidal synergy at approximately 10(5) CFU/ml of inoculum (but not at approximately 10(7)-CFU inoculum), whereas other beta-lactamase inhibitors (sulbac...

  3. Dissemination of the novel plasmid-mediated beta-lactamase CTX-1, which confers resistance to broad-spectrum cephalosporins, and its inhibition by beta-lactamase inhibitors.

    OpenAIRE

    Kitzis, M D; Billot-Klein, D; Goldstein, F W; Williamson, R.; Tran Van Nhieu, G; Carlet, J; Acar, J F; Gutmann, L

    1988-01-01

    The novel beta-lactamase CTX-1 (pI 6.3) encoded on a transferable 84-kilobase plasmid was found in six different bacterial species. It was responsible for a significant decrease in susceptibility towards most penicillins and cephalosporins, except imipenem, temocillin, and cephalosporins which have a 7-alpha-methoxy substituent. Synergy between either ampicillin, piperacillin, cefotaxime, ceftazidime, or aztreonam and three beta-lactamase inhibitors (clavulanic acid, sulbactam, and YTR 830) w...

  4. Thermodynamic Cycle Analysis and Inhibitor Design against Beta-Lactamase

    Energy Technology Data Exchange (ETDEWEB)

    Roth, Tomer A.; Minasov, George; Morandi, Stefania; Prati, Fabio; Shoichet, Brian K. (Degali); (UCSF)

    2010-03-08

    {beta}-Lactamases are the most widespread resistance mechanism to {beta}-lactam antibiotics, such as the penicillins and cephalosporins. Transition-state analogues that bind to the enzymes with nanomolar affinities have been introduced in an effort to reverse the resistance conferred by these enzymes. To understand the origins of this affinity, and to guide design of future inhibitors, double-mutant thermodynamic cycle experiments were undertaken. An unexpected hydrogen bond between the nonconserved Asn289 and a key inhibitor carboxylate was observed in the X-ray crystal structure of a 1 nM inhibitor (compound 1) in complex with AmpC {beta}-lactamase. To investigate the energy of this hydrogen bond, the mutant enzyme N289A was made, as was an analogue of 1 that lacked the carboxylate (compound 2). The differential affinity of the four different protein and analogue complexes indicates that the carboxylate-amide hydrogen bond contributes 1.7 kcal/mol to overall binding affinity. Synthesis of an analogue of 1 where the carboxylate was replaced with an aldehyde led to an inhibitor that lost all this hydrogen bond energy, consistent with the importance of the ionic nature of this hydrogen bond. To investigate the structural bases of these energies, X-ray crystal structures of N289A/1 and N289A/2 were determined to 1.49 and 1.39 {angstrom}, respectively. These structures suggest that no significant rearrangement occurs in the mutant versus the wild-type complexes with both compounds. The mutant enzymes L119A and L293A were made to investigate the interaction between a phenyl ring in 1 and these residues. Whereas deletion of the phenyl itself diminishes affinity by 5-fold, the double-mutant cycles suggest that this energy does not come through interaction with the leucines, despite the close contact in the structure. The energies of these interactions provide key information for the design of improved inhibitors against {beta}-lactamases. The high magnitude of the ion

  5. Correlation between beta-lactamase production and MIC values against penicillin with coagulase negative staphylococci.

    Directory of Open Access Journals (Sweden)

    Narayani T

    1989-07-01

    Full Text Available Two hundred strains of coagulase negative staphylococci (CNS isolated from various clinical specimens (116 and healthy hospital personnel (84 were investigated for the production of beta-lactamases by means of three iodometric techniques and correlated with the minimum inhibitory concentration (MIC values of penicillin-G by agar dilution technique and disc diffusion technique. One hundred and fifty (75.0% of the 200 strains tested produced beta-lactamases. Seventy two per cent of the CNS were found to be beta-lactamase positive by the starch paper technique which was the most sensitive one in our study. The MIC values of penicillin against CNS ranged from less than or equal to 1.25 to greater than or equal to 2000 units. The present study indicated the higher prevalence of beta-lactamase producers with increased penicillin resistance among CNS strains isolated from healthy carriers and hospitalised patients.

  6. Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    Bruno F. Robles

    2014-04-01

    Full Text Available The objectives of the study were to evaluate the presence/production of beta-lactamases by both phenotypic and genotypic methods, verify whether results are dependent of bacteria type (Staphylococcus aureus versus coagulase-negative Staphylococcus - CNS and verify the agreement between tests. A total of 200 bacteria samples from 21 different herds were enrolled, being 100 CNS and 100 S. aureus. Beta-lactamase presence/detection was performed by different tests (PCR, clover leaf test - CLT, Nitrocefin disk, and in vitro resistance to penicillin. Results of all tests were not dependent of bacteria type (CNS or S. aureus. Several S. aureus beta-lactamase producing isolates were from the same herd. Phenotypic tests excluding in vitro resistance to penicillin showed a strong association measured by the kappa coefficient for both bacteria species. Nitrocefin and CLT are more reliable tests for detecting beta-lactamase production in staphylococci.

  7. BetalasEN: microdilution panel for identifying beta-lactamases present in isolates of Enterobacteriaceae.

    Science.gov (United States)

    Sanders, Christine C; Ehrhardt, Anton F; Moland, Ellen Smith; Thomson, Kenneth S; Zimmer, Barbara; Roe, Darcie E

    2002-01-01

    A dried investigational use-only microdilution panel named betalasEN (a short named derived from the panel's purpose, to identify beta-lactamases in Enterobacteriaceae) containing 10 beta-lactam drugs with and without beta-lactamase inhibitors was developed to identify beta-lactamases among clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter koseri, Citrobacter freundii group, Enterobacter spp., and Serratia marcescens. The MICs obtained with a collection of 383 organisms containing well-characterized beta-lactamases were used to develop numeric codes and logic pathways for computerized analysis of results. The resultant logic pathways and betalasEN panel were then used to test and identify beta-lactamases among 885 isolates of Enterobacteriaceae recovered in cultures obtained at six different hospital laboratories across the United States. beta-Lactamases present in 801 (90.5%) of the 885 isolates were identified by betalasEN by using the existing logic pathways and codes or after minor modifications were made to the existing codes. The 84 strains that gave codes that betalasEN could not identify were collected, reidentified, and retested by using betalasEN. Three strains had been misidentified, 54 strains gave different codes upon repeat testing that could be identified by betalasEN, and 27 strains repeated new codes. The beta-lactamases in these strains were identified, and the new codes were added to the betalasEN logic pathways. These results indicate that betalasEN can identify clinically important beta-lactamases among most isolates of Enterobacteriaceae. The results also show that good quality control and attention to proper performance of the tests are essential to the correct performance of betalasEN. PMID:11773104

  8. Evolutionary Trajectories of Beta-Lactamase CTX-M-1 Cluster Enzymes: Predicting Antibiotic Resistance

    OpenAIRE

    Angela Novais; Iñaki Comas; Fernando Baquero; Rafael Cantón; Coque, Teresa M.; Andrés Moya; Fernando González-Candelas; Juan-Carlos Galán

    2010-01-01

    Extended-spectrum beta-lactamases (ESBL) constitute a key antibiotic-resistance mechanism affecting Gram-negative bacteria, and also an excellent model for studying evolution in real time. A shift in the epidemiology of ESBLs is being observed, which is characterized by the explosive diversification and increase in frequency of the CTX-M-type beta-lactamases in different settings. This provides a unique opportunity for studying a protein evolutionary radiation by the sequential acquisition of...

  9. Correlation between beta-lactamase production and MIC values against penicillin with coagulase negative staphylococci.

    OpenAIRE

    Narayani T; Shanmugam J; Naseema K; Bhattacharya R; Shyamkrishnan K

    1989-01-01

    Two hundred strains of coagulase negative staphylococci (CNS) isolated from various clinical specimens (116) and healthy hospital personnel (84) were investigated for the production of beta-lactamases by means of three iodometric techniques and correlated with the minimum inhibitory concentration (MIC) values of penicillin-G by agar dilution technique and disc diffusion technique. One hundred and fifty (75.0%) of the 200 strains tested produced beta-lactamases. Seventy two per cent of ...

  10. Surveillance and Detection of Inhibitor-Resistant Beta-Lactamases in Clinical Isolates of Escherichia coli

    OpenAIRE

    Carl Urban; Patricia Bradford; Joseph Villarin

    2007-01-01

    In recent years, resistance to beta-lactam antibiotics, such as the widely-used cephalosporins and penicillins, has become a major challenge for disease therapy, particularly in common hospital-acquired infections. In the search for the mechanisms behind this increasingly prevalent form of resistance, microbiologists have identified a new type of beta-lactamase enzyme, called inhibitor-resistant TEMs (IRTs), which can withstand the effects of beta-lactamase inhibitor compounds, further reduci...

  11. Penicillin-binding proteins and induction of AmpC beta-lactamase.

    OpenAIRE

    Sanders, C C; Bradford, P A; Ehrhardt, A F; Bush, K; Young, K D; Henderson, T A; Sanders, W E

    1997-01-01

    In competition assays for radiolabeled penicillin, penicillin-binding proteins (PBPs) 4, 7a, and 7b showed very high affinities for strong inducers of AmpC beta-lactamase. Loss of PBP 4 resulted in diminished inducibility. This suggests that if PBPs are involved in induction of AmpC beta-lactamase, there is probably a redundancy in function among the different PBPs.

  12. Plasmid-mediated quinolone resistance in expanded spectrum beta lactamase producing enterobacteriaceae in Morocco.

    OpenAIRE

    Bouchakour, Mohammed; Zerouali, Khalid; Gros Claude, Jean David Perrier; Amarouch, Hamid; El Mdaghri, Naima; Courvalin, Patrice; Timinouni, Mohammed

    2010-01-01

    INTRODUCTION: Although independently acquired, plasmid-mediated quinolone resistance appears to be linked with extended-spectrum or AmpC-type beta-lactamases. Since no data are available in African countries, the prevalence of qnr genes at the University Hospital Ibn Rochd, Casablanca, Morocco, was investigated. METHODOLOGY: Between October 2006 and March 2007, the following 39 randomly selected non-duplicate Enterobacteriaceae producing an extended-spectrum beta-lactamase (ESBL), representin...

  13. Chromosomal beta-lactamase is packaged into membrane vesicles and secreted from Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Ciofu, O; Beveridge, T J; Kadurugamuwa, J; Walther-Rasmussen, J; Høiby, N

    2000-01-01

    Membrane vesicles were isolated from one beta-lactam-sensitive and three beta-lactam-resistant Pseudomonas aeruginosa clinical isolates from patients with cystic fibrosis. The presence of the chromosomally encoded beta-lactamase in the membrane vesicles was shown by electron microscopy and...... enzymatic studies. This is the first report of extracellular secretion of beta-lactamase in P. aeruginosa and it seems that the enzyme is packaged into membrane vesicles....

  14. Physiological studies of the regulation of beta-lactamase expression in Pseudomonas maltophilia.

    Science.gov (United States)

    Rosta, S; Mett, H

    1989-01-01

    The kinetics of beta-lactamase induction in Pseudomonas maltophilia IID1275/873 were investigated. Upon induction with beta-lactam antibiotics, a correlation was seen between the increase in specific beta-lactamase activity and the generation time, as well as the concentration of inducer in the medium. The specific beta-lactamase activity increased slowly within the first 0.5 generation and then more rapidly; it decreased regularly after about 2 generations of growth in the presence of inducer. This decrease could presumably be attributed to the continuous breakdown of inducer by beta-lactamases in the culture medium. In a chemostat culture with continuous supply of fresh inducer-containing medium, the specific beta-lactamase activity could be stabilized at a high level over several generations. Removal of the beta-lactam after a certain induction time showed that a short exposure of the bacteria to inducer caused induction kinetics comparable to those resulting from continuous exposure of the cells to inducer. The two beta-lactamases of P. maltophilia, L1 and L2, were induced simultaneously under various experimental conditions. PMID:2783690

  15. Beta-lactamase Escherichia coli and Staphylococcus aureus isolated from chickens in Nigeria

    Directory of Open Access Journals (Sweden)

    Godwin Onyemaechi Egwu

    2010-06-01

    Full Text Available The occurrence of beta-lactamase-producing Escherichia coli and Staphylococcus aureus in chickens was investigated. Specimens (n = 1 300 were collected from 400 chickens and were streaked on MacConkey agar plates. From each plate, presumptive growths of organisms were picked and streaked on eosin methylene blue and Baird-Parker agars, respectively. Typical colonies of E. coli and S. aureus with similar morphologies were identified by biochemical tests. Isolates were tested for beta-lactamase production and antimicrobial susceptibilities. Results indicated that 805 E. coli isolates from which 89 (11% were beta-lactamase-positive and 660 S. aureus from which 58 (8.8% were beta-lactamase-positive. Both isolates showed a high level of resistance to all twelve antibiotics screened. The increased prevalence of antibiotic resistance amongst bacterial organisms is undoubtedly correlated with the discovery and characterisation of multiple, transferrable resistance determinants, such as beta-lactamases, corresponding to their respective phenotypes. The implications of this for humans when handling and/or consuming chickens and chicken products contaminated with strains of such isolates, is a risk of transferrable multi-drug resistance and a failure of treatment. The results of our study indicated that beta-lactamase-producing E. coli and S. aureus are prevalent in chickens in Nigeria.

  16. Beta-lactamase Escherichia coli and Staphylococcus aureus isolated from chickens in Nigeria.

    Science.gov (United States)

    Mamza, Sunday Akidarju; Egwu, Godwin Onyemaechi; Mshelia, Gideon Dauda

    2010-01-01

    The occurrence of beta-lactamase-producing Escherichia coli and Staphylococcus aureus in chickens was investigated. Specimens (n = 1,300) were collected from 400 chickens and were streaked on MacConkey agar plates. From each plate, presumptive growths of organisms were picked and streaked on eosin methylene blue and Baird-Parker agars, respectively. Typical colonies of E. coli and S. aureus with similar morphologies were identified by biochemical tests. Isolates were tested for beta-lactamase production and antimicrobial susceptibilities. Results indicated that 805 E. coli isolates from which 89 (11%) were beta-lactamase-positive and 660 S. aureus from which 58 (8.8%) were beta-lactamase-positive. Both isolates showed a high level of resistance to all twelve antibiotics screened. The increased prevalence of antibiotic resistance amongst bacterial organisms is undoubtedly correlated with the discovery and characterisation of multiple, transferrable resistance determinants, such as beta-lactamases, corresponding to their respective phenotypes. The implications of this for humans when handling and/or consuming chickens and chicken products contaminated with strains of such isolates, is a risk of transferrable multi-drug resistance and a failure of treatment. The results of our study indicated that beta-lactamase-producing E. coli and S. aureus are prevalent in chickens in Nigeria. PMID:20560125

  17. Exposure assessment of extended-spectrum beta-lactamases/AmpC beta-lactamases-producing Escherichia coli in meat in Denmark

    OpenAIRE

    Carmo, Luís P.; Liza R. Nielsen; da Costa, Paulo M.; Alban, Lis

    2014-01-01

    Introduction: Extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases (AmpC) are of concern for veterinary and public health because of their ability to cause treatment failure due to antimicrobial resistance in Enterobacteriaceae. The main objective was to assess the relative contribution (RC) of different types of meat to the exposure of consumers to ESBL/AmpC and their potential importance for human infections in Denmark.Material and methods: The prevalence of each genotype of ES...

  18. Use of microdilution panels with and without beta-lactamase inhibitors as a phenotypic test for beta-lactamase production among Escherichia coli, Klebsiella spp., Enterobacter spp., Citrobacter freundii, and Serratia marcescens.

    Science.gov (United States)

    Thomson, K S; Sanders, C C; Moland, E S

    1999-06-01

    Over the past decade, a number of new beta-lactamases have appeared in clinical isolates of Enterobacteriaceae that, unlike their predecessors, do not confer beta-lactam resistance that is readily detected in routine antibiotic susceptibility tests. Because optimal methodologies are needed to detect these important new beta-lactamases, a study was designed to evaluate the ability of a panel of various beta-lactam antibiotics tested alone and in combination with beta-lactamase inhibitors to discriminate between the production of extended-spectrum beta-lactamases, AmpC beta-lactamases, high levels of K1 beta-lactamase, and other beta-lactamases in 141 isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, and Serratia marcescens possessing well-characterized beta-lactamases. The microdilution panels studied contained aztreonam, cefpodoxime, ceftazidime, cefotaxime, and ceftriaxone, with and without 1, 2, and 4 microg of clavulanate per ml or 8 microg of sulbactam per ml and cefoxitin and cefotetan with and without 8 microg of sulbactam per ml. The results indicated that a minimum panel of five tests would provide maximum separation of extended-spectrum beta-lactamase high AmpC, high K1, and other beta-lactamase production in Enterobacteriaceae. These included cefpodoxime, cefpodoxime plus 4 microg of clavulanate per ml, ceftazidime, ceftriaxone, and ceftriaxone plus 8 microg of sulbactam per ml. Ceftriaxone plus 2 microg of clavulanate per ml could be substituted for cefpodoxime plus 4 microg of clavulanate per ml without altering the accuracy of the tests. This study indicated that tests with key beta-lactam drugs, alone and in combination with beta-lactamase inhibitors, could provide a convenient approach to the detection of a variety of beta-lactamases in members of the family Enterobacteriaceae. PMID:10348759

  19. Detection of genes mediating beta-lactamase production in isolates of enterobacteria recovered from wild pets in Saudi Arabia

    OpenAIRE

    Sabry A. Hassan; Mohamed Y. Shobrak

    2015-01-01

    Aim: To determine the genetic basis and types of beta-lactamase encountered among enterobacterial isolates of wild pets from the animal exhibit. Materials and Methods: A total of 17 beta-lactamase-producing enterobacteria recovered from fecal samples of wild pet animals were analyzed for a selected beta-lactamase gene by polymerase chain reaction. Results: Molecular analysis identified one or more β-lactamase-encoding genes in 14 enterobacterial isolates as a single or gene combination....

  20. Frequency and Antibiotic Susceptibility Pattern of Amp C Beta Lactamase Producing Bacteria Isolated from a Tertiary Care Hospital of Pakistan

    OpenAIRE

    Afreenish Hassan; Javaid Usman; Fatima Kaleem; Maria Omair; Ali Khalid; Muhammad Iqabal

    2011-01-01

    Objective: Amp C beta lactamases are cephalosporinases which hydrolyze cephamycins and are poorlyinhibited by clavulanic acid. Amp C beta lactamases confer resistance to a wide variety of antibiotics andpose both diagnostic and therapeutic challenges, The objective was to detect the frequency and antibioticsusceptibility pattern of Amp C beta lactamase producing bacteria isolated from a tertiary care hospital ofPakistan.Methodology: Organisms were isolated from various clinical specimens. Fir...

  1. Detection and characterization of metallo beta lactamases producing Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Manoharan A

    2010-01-01

    Full Text Available This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs among nosocomial Pseudomonas aeruginosa. Sixty one among 176 P. aeruginosa isolates, collected as part of a multicentric study (2005-2007, were evaluated for carbapenem resistance (CARB-R; resistant to either imipenem/meropenem and screened for MBL by Combination Disk Diffusion Test (CDDT using imipenem (IMP, meropenem (MER and ceftazidime (CAZ with EDTA. MBL positives were further confirmed by IMP + EDTA Etest. Twenty strains (42.6% were found to be MBL producers among the 61 P. aeruginosa. PCR for IMP and VIM MBL was performed on 48 of the 61, 15 were positive for VIM MBL type. CDDT using IMP + EDTA had the highest sensitivity and specificity of 87.8% and 84.4% when compared to Etest, which was higher than the values obtained for CAZ + EDTA and MER + EDTA. CDDT using IMP + EDTA also compared very well with the PCR (specificity = 90.9%, sensitivity = 93.3%. CARB-R among P. aeruginosa is mediated predominantly via MBL production. Clinical P. aeruginosa isolates can be screened routinely using the less expensive IMP + EDTA CDDT in clinical microbiology laboratories.

  2. METALLO-BETA-LACTAMASE PRODUCING PSEUDOMONAS AERUGINOSA IN NEONATAL SEPTICEMIA

    Directory of Open Access Journals (Sweden)

    Murthy

    2014-05-01

    Full Text Available The emergence, selective multiplication & dissemination of antibacterial resistance is a serious global problem. This study was conducted with the objective to examine the incidence of metallo-beta-lactamase (MβL producing strains among multidrug resistant (MDR Pseudomonas aeruginosa from the suspected cases of neonatal sepsis between January 2011 – December 2013. A total of 994 cases admitted with the suspicion of neonatal sepsis were investigated. 295 (29.7% isolates were obtained from the blood cultures of neonates. The isolates were identified and tested for the susceptibility to various antimicrobial agents. Pseudomonas aeruginosa with 116 (48.3% isolation among 240 Gram negative isolates, was the predominant pathogen in our study. All the 74 (63.8% multidrug resistant P. aeruginosa isolates were screened initially for Imipenem resistance, which were further tested for the presence of MβL by Imipenem-ethylene diamine tetraacetic acid (EDTA disc method. MβL production was seen in 20 (71.4% of the 28 Imipenem-resistant Pseudomonas aeruginosa isolates. MβL producing Pseudomonas aeruginosa has emerged as a potential threat in cases of neonatal septicemia and poses great therapeutic challenge for physicians treating such infections.

  3. Beta-lactamases among Extended spectrum Beta-lactamase resistant (ESBL) Salmonella from poultry, poultry products and human patients in The Netherlands

    DEFF Research Database (Denmark)

    Hasman, Henrik; Mevius, D.; Veldman, K.;

    2006-01-01

    Objectives: The purpose of this work was to study the genetic determinants responsible for extended-spectrum beta-lactamase (ESBL) resistance of Salmonella isolated from Dutch poultry, poultry meat and hospitalized humans. Methods: Thirty-four ESBL-resistant Salmonella isolates from The Netherlands...... enterica isolated from animals, food products and human patients....

  4. beta-Lactamases among extended-spectrum beta-lactamase (ESBL)-resistant Salmonella from poultry, poultry products and human patients in The Netherlands

    DEFF Research Database (Denmark)

    Hasman, Henrik; Mevius, D.; Veldman, K.;

    2005-01-01

    Objectives: The purpose of this work was to study the genetic determinants responsible for extended-spectrum beta-lactamase (ESBL) resistance of Salmonella isolated from Dutch poultry, poultry meat and hospitalized humans. Methods: Thirty-four ESBL-resistant Salmonella isolates from The Netherlands...... enterica isolated from animals, food products and human patients....

  5. Beta-Lactamases among extended-spectrum (beta)-lactamase (ESBL)-resistant Salmonella from poultry, poultry products and human patients in The Netherlands

    NARCIS (Netherlands)

    Hasman, H.; Mevius, D.J.; Veldman, K.T.; Olesen, I.; Aarestrup, F.M.

    2005-01-01

    Objectives: The purpose of this work was to study the genetic determinants responsible for extended-spectrum beta-lactamase (ESBL) resistance of Salmonella isolated from Dutch poultry, poultry meat and hospitalized humans. Methods: Thirty-four ESBL-resistant Salmonella isolates from The Netherlands

  6. CLONING, SEQUENCE ANALYSIS, AND CHARACTERIZATION OF PUTATIVE BETA-LACTAMASE OF STENOTROPHOMONAS MALTOPHILIA

    Directory of Open Access Journals (Sweden)

    Chong Seng Shueh

    2012-10-01

    Full Text Available The main objective of current study was to explore the function of chromosomal putative beta-lactamase gene (smlt 0115 in clinical Stenotrophomonas maltophilia. Antibiotic susceptibility test (AST screening for current antimicrobial drugs was done and Minimum Inhibitory Concentration (MIC level towards beta-lactams was determined by E-test. Putative beta-lactamase gene of S. maltophilia was amplified via PCR, with specific primers, then cloned into pET-15 expression plasmid and transformed into Escherichia coli BL21. The gene was sequenced and analyzed. The expressed protein was purified by affinity chromatography and the kinetic assay was performed. S. maltophilia ATCC 13637 was included in this experiment. Besides, a hospital strain which exhibited resistant to a series of beta-lactams including cefepime was identified via AST and MIC, hence it was named as S2 strain and was considered in this study. Sequencing result showed that putative beta-lactamase gene obtained from ATCC 13637 and S2 strains were predicted to have cephalosporinase activity by National Center for Biotechnology Information (NCBI blast program. Differences in the sequences of both ATCC 13637 and S2 strains were found via ClustalW alignment software. Kinetic assay proved a cephalosporinase characteristic produced by E. coli BL21 clone that overexpressed the putative beta-lactamase gene cloned under the control of an external promoter. Yet, expressed protein purified from S2 strain had high catalytic activity against beta-lactam antibiotics which was 14-fold higher than expressed protein purified from ATCC 13637 strain. This study represents the characterization analysis of putative beta-lactamase gene (smlt 0115 of S. maltophilia. The presence of the respective gene in the chromosome of S. maltophilia suggested that putative beta-lactamase gene (smlt 0115 of S. maltophilia plays a role in beta-lactamase resistance.

  7. The in vitro activity of beta-lactamase inhibitors in combination with cephalosporins against M. tuberculosis.

    Science.gov (United States)

    Chen, C H; Yang, M H; Lin, J S; Lee, Y C; Perng, R P

    1995-04-01

    Although there are reports that the addition of a beta-lactamase inhibitor to ampicillin or amoxicillin greatly improves their in vitro activity against M. tuberculosis, there are no written reports about the antituberculosis effects of beta-lactamase inhibitors in combination with cephalosporins against M. tuberculosis. In this report, we have determined the minimal inhibitory concentrations (MIC) of 5 cephalosporins with or without combination with beta-lactamase inhibitor against M. tuberculosis strains isolated from patients before antituberculosis treatment and checked the production of beta-lactamase by bacteria before this procedure. Four strains of M. tuberculosis were contaminated during the experiment, and all the other 16 strains hydrolyzed the nitrocefin disc, thus indicating a beta-lactamase producer. The MICs of cephalosporins alone against M. tuberculosis were 200-400 micrograms/ml for ceforanide, 100-400 micrograms/ml for cephapirin, 400-1600 micrograms/ml for cefamandole, 200-1600 micrograms/ml for cefotaxime, and 800-1600 micrograms/ml for ceftriaxone. After adding the equimolar concentrations of sulbactam, the MICs were reduced to 100-200 micrograms/ml for ceforanide, 12.5-100 micrograms/ml for cephapirin, 100-400 micrograms/ml for cefamandole, 25-200 micrograms/ml for cefotaxime, and 100-800 micrograms/ml for ceftriaxone. We concluded that sulbactam enhanced the antituberculosis effect of cephalosporins. PMID:7624446

  8. Effect of porin loss on the activity of tigecycline against Klebsiella pneumoniae producing extended-spectrum beta-lactamases or plasmid-mediated AmpC-type beta-lactamases.

    Science.gov (United States)

    Conejo, M Carmen; Hernández, J Ramón; Pascual, Alvaro

    2008-07-01

    Tigecycline showed excellent in vitro activity against 50 clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamases, plasmid-mediated AmpC-type beta-lactamases, or both. This activity was not affected by porin loss. Porin loss, however, did affect the activity of imipenem against strains that expressed both types of enzymes. PMID:18339509

  9. Pseudomonas aeruginosa isolates from patients with cystic fibrosis have different beta-lactamase expression phenotypes but are homogeneous in the ampC-ampR genetic region

    DEFF Research Database (Denmark)

    Campbell, J I; Ciofu, O; Høiby, N

    1997-01-01

    Pseudomonas aeruginosa isolates from 1 of 17 cystic fibrosis patients produced secondary beta-lactamase in addition to the ampC beta-lactamase. Isolates were grouped into three beta-lactamase expression phenotypes: (i) beta-lactam sensitive, low basal levels and inducible beta-lactamase production......; (ii) beta-lactam resistant, moderate basal levels and hyperinducible beta-lactamase production; (iii) beta-lactam resistant, high basal levels and constitutive beta-lactamase production. Apart from a base substitution in the ampR-ampC intergenic region of an isolate with moderate-basal-level and...... hyperinducible beta-lactamase production, sensitive and resistant strains were identical in their ampC-ampR genetic regions. Thus, enhanced beta-lactamase expression is due to mutations in regulatory proteins other than AmpR....

  10. The prevalence of extended-spectrum beta-lactamase in environmental isolates of Enterobacter.

    Science.gov (United States)

    Sharma, Anjana; Dour, Prashant; Singh, Thakur Nirbhay

    2008-01-01

    The incidence of extended-spectrum beta-lactamase (ESBL)-producing strains and multidrug-resistant strains of Enterobacter spp. isolated from the 1312 km long river Narmada was investigated. Out of the 57 isolates of Enterobacter, 73.68% were found to be ESBL producers including the isolates of E. taylorae and isolates of E. agglomerans, which have been characterized for the first time. All the isolates were found susceptible to the antibiotic imipenem. AmpC gene was found in all the Enterobacter strains tested. AmpC beta-lactamase-producing bacterial pathogens may cause major therapeutic failure if not detected and reported in time. It was seen that these enzymes are mainly chromosomally mediated along with several non-AmpC beta-lactamase. PMID:18417885

  11. Using steric hindrance to design new inhibitors of class C beta-lactamases

    Energy Technology Data Exchange (ETDEWEB)

    Trehan, Indi; Morandi, F.; Blaszczak, L.C.; Shoichet, Brian K. (NWU)

    2010-03-08

    {beta}-lactamases confer resistance to {beta}-lactam antibiotics such as penicillins and cephalosporins. However, {beta}-lactams that form an acyl-intermediate with the enzyme but subsequently are hindered from forming a catalytically competent conformation seem to be inhibitors of {beta}-lactamases. This inhibition may be imparted by specific groups on the ubiquitous R1 side chain of {beta}-lactams, such as the 2-amino-4-thiazolyl methoxyimino (ATMO) group common among third-generation cephalosporins. Using steric hindrance of deacylation as a design guide, penicillin and carbacephem substrates were converted into effective {beta}-lactamase inhibitors and antiresistance antibiotics. To investigate the structural bases of inhibition, the crystal structures of the acyl-adducts of the penicillin substrate amoxicillin and the new analogous inhibitor ATMO-penicillin were determined. ATMO-penicillin binds in a catalytically incompetent conformation resembling that adopted by third-generation cephalosporins, demonstrating the transferability of such sterically hindered groups in inhibitor design.

  12. New variant of TEM-10 beta-lactamase gene produced by a clinical isolate of proteus mirabilis.

    OpenAIRE

    Palzkill, T.; Thomson, K S; Sanders, C C; Moland, E S; Huang, W.; Milligan, T W

    1995-01-01

    A clinical isolate of Proteus mirabilis was found to produce a new variant of the TEM-10 beta-lactamase gene. This is the first report of TEM-10 production by P. mirabilis and the first report of extended-spectrum beta-lactamase production by an isolate of this species recovered in the United States.

  13. Novel Computational Protocols for Functionally Classifying and Characterising Serine Beta-Lactamases.

    Science.gov (United States)

    Lee, David; Das, Sayoni; Dawson, Natalie L; Dobrijevic, Dragana; Ward, John; Orengo, Christine

    2016-06-01

    Beta-lactamases represent the main bacterial mechanism of resistance to beta-lactam antibiotics and are a significant challenge to modern medicine. We have developed an automated classification and analysis protocol that exploits structure- and sequence-based approaches and which allows us to propose a grouping of serine beta-lactamases that more consistently captures and rationalizes the existing three classification schemes: Classes, (A, C and D, which vary in their implementation of the mechanism of action); Types (that largely reflect evolutionary distance measured by sequence similarity); and Variant groups (which largely correspond with the Bush-Jacoby clinical groups). Our analysis platform exploits a suite of in-house and public tools to identify Functional Determinants (FDs), i.e. residue sites, responsible for conferring different phenotypes between different classes, different types and different variants. We focused on Class A beta-lactamases, the most highly populated and clinically relevant class, to identify FDs implicated in the distinct phenotypes associated with different Class A Types and Variants. We show that our FunFHMMer method can separate the known beta-lactamase classes and identify those positions likely to be responsible for the different implementations of the mechanism of action in these enzymes. Two novel algorithms, ASSP and SSPA, allow detection of FD sites likely to contribute to the broadening of the substrate profiles. Using our approaches, we recognise 151 Class A types in UniProt. Finally, we used our beta-lactamase FunFams and ASSP profiles to detect 4 novel Class A types in microbiome samples. Our platforms have been validated by literature studies, in silico analysis and some targeted experimental verification. Although developed for the serine beta-lactamases they could be used to classify and analyse any diverse protein superfamily where sub-families have diverged over both long and short evolutionary timescales. PMID

  14. Inhibition of AmpC beta-lactamase through a destabilizing interaction in the active site

    Energy Technology Data Exchange (ETDEWEB)

    Trehan, I.; Beadle, B.M.; Shoichet, B.K. (NWU)

    2010-03-08

    {beta}-Lactamases hydrolyze {beta}-lactam antibiotics, including penicillins and cephalosporins; these enzymes are the most widespread resistance mechanism to these drugs and pose a growing threat to public health. {beta}-Lactams that contain a bulky 6(7){alpha} substituent, such as imipenem and moxalactam, actually inhibit serine {beta}-lactamases and are widely used for this reason. Although mutant serine {beta}-lactamases have arisen that hydrolyze {beta}-lactamase resistant {beta}-lactams (e.g., ceftazidime) or avoid mechanism-based inhibitors (e.g., clavulanate), mutant serine {beta}-lactamases have not yet arisen in the clinic with imipenemase or moxalactamase activity. Structural and thermodynamic studies suggest that the 6(7){alpha} substituents of these inhibitors form destabilizing contacts within the covalent adduct with the conserved Asn152 in class C {beta}-lactamases (Asn132 in class A {beta}-lactamases). This unfavorable interaction may be crucial to inhibition. To test this destabilization hypothesis, we replaced Asn152 with Ala in the class C {beta}-lactamase AmpC from Escherichia coli and examined the mutant enzyme's thermodynamic stability in complex with imipenem and moxalactam. Consistent with the hypothesis, the Asn152 {yields} Ala substitution relieved 0.44 and 1.10 kcal/mol of strain introduced by imipenem and moxalactam, respectively, relative to the wild-type complexes. However, the kinetic efficiency of AmpC N152A was reduced by 6300-fold relative to that of the wild-type enzyme. To further investigate the inhibitor's interaction with the mutant enzyme, the X-ray crystal structure of moxalactam in complex with N152A was determined to a resolution of 1.83 {angstrom}. Moxalactam in the mutant complex is significantly displaced from its orientation in the wild-type complex; however, moxalactam does not adopt an orientation that would restore competence for hydrolysis. Although Asn152 forces {beta}-lactams with 6(7){alpha

  15. Energetic, Structural, and Antimicrobial Analyses of [beta]-Lactam Side Chain Recognition by [beta]-Lactamases

    Energy Technology Data Exchange (ETDEWEB)

    Caselli, E.; Powers, R.A.; Blaszczak, L.C.; Wu, C.Y.E.; Prati, F.; Shoichet, B.K. (NWU)

    2010-03-05

    Penicillins and cephalosporins are among the most widely used and successful antibiotics. The emergence of resistance to these {beta}-lactams, most often through bacterial expression of {beta}-lactamases, threatens public health. To understand how {beta}-lactamases recognize their substrates, it would be helpful to know their binding energies. Unfortunately, these have been difficult to measure because {beta}-lactams form covalent adducts with {beta}-lactamases. This has complicated functional analyses and inhibitor design. To investigate the contribution to interaction energy of the key amide (R1) side chain of {beta}-lactam antibiotics, eight acylglycineboronic acids that bear the side chains of characteristic penicillins and cephalosporins, as well as four other analogs, were synthesized. These transition-state analogs form reversible adducts with serine {beta}-lactamases. Therefore, binding energies can be calculated directly from K{sub i} values. The K{sub i} values measured span four orders of magnitude against the Group I {beta}-lactamase AmpC and three orders of magnitude against the Group II {beta}-lactamase TEM-1. The acylglycineboronic acids have K{sub i} values as low as 20 nM against AmpC and as low as 390 nM against TEM-1. The inhibitors showed little activity against serine proteases, such as chymotrypsin. R1 side chains characteristic of {beta}-lactam inhibitors did not have better affinity for AmpC than did side chains characteristic of {beta}-lactam substrates. Two of the inhibitors reversed the resistance of pathogenic bacteria to {beta}-lactams in cell culture. Structures of two inhibitors in their complexes with AmpC were determined by X-ray crystallography to 1.90 {angstrom} and 1.75 {angstrom} resolution; these structures suggest interactions that are important to the affinity of the inhibitors. Acylglycineboronic acids allow us to begin to dissect interaction energies between {beta}-lactam side chains and {beta}-lactamases. Surprisingly

  16. Characterization of a novel extended-spectrum beta-lactamase from Pseudomonas aeruginosa.

    OpenAIRE

    Nordmann, P.; Ronco, E; Naas, T.; Duport, C; Michel-Briand, Y.; Labia, R

    1993-01-01

    A clinical isolate of Pseudomonas aeruginosa RNL-1 showed resistance to extended-spectrum cephalosporins which was inhibited by clavulanic acid. Although this strain contained three plasmids ca. 80, 20, and 4 kb long, the resistance could not be transferred by mating-out assays with P. aeruginosa or Escherichia coli. Cloning of a 2.1-kb Sau3A fragment from P. aeruginosa RNL-1 into plasmid pACYC184 produced pPZ1, a recombinant plasmid that encodes a beta-lactamase. This beta-lactamase (PER-1) ...

  17. Occurrence and characteristics of extended spectrum beta-lactamases-producing Enterobacteriaceae from foods of animal origin.

    Science.gov (United States)

    Tekiner, İsmail Hakkı; Özpınar, Haydar

    2016-01-01

    Presence of extended spectrum beta-lactamases (ESBL) in bacteria is a growing health concern of global significance. The local, regional, national, and international epidemiological studies for extended spectrum beta-lactamases-producing Enterobacteriaceae and their encoding genes in foods are still incomplete. The objective of this study was to determine the occurrence of extended spectrum beta-lactamases-producing Enterobacteriaceae and the characteristics of their encoding genes from a total of 250 samples of various foods of animal-origin (100 raw chicken meat, 100 raw cow milk, and 50 raw cow milk cheese) sold in Turkey. Overall, 55 isolates were positive as extended spectrum beta-lactamases-producing Enterobacteriaceae. The most prevalent extended spectrum beta-lactamases-producing strain were identified as Escherichia coli (80%), followed by Enterobacter cloacae (9.1%), Citrobacter braakii (5.5%), Klebsiella pneumoniae (3.6%), and Citrobacter werkmanii (1.8%) by Vitek(®) MS. The simultaneous production of extended spectrum beta-lactamases and AmpC was detected in five isolates (9.1%) in E. coli (80%) and E. cloacae (20%). The frequency rates of blaTEM, blaCTX-M, and blaSHV were 96.4%, 53.7%, and 34.5%, respectively. The co-existence of bla-genes was observed in 82% of extended spectrum beta-lactamases producers with a distribution of blaTEM &blaCTX-M (52.7%), blaTEM &blaSHV (20%), blaTEM &blaCTX-M &blaSHV (12.7%), and blaSHV &blaCTX-M (1.8%). The most prevalent variant of blaCTX-M clusters was defined as blaCTX-M-1 (97.2%), followed by blaCTX-M-8 (2.8%). In summary, the analysed foods were found to be posing a health risk for Turkish consumers due to contamination by Enterobacteriaceae with a diversity of extended spectrum beta-lactamases encoding genes. PMID:26991276

  18. Terminal truncations in amp C beta-lactamase from a clinical isolate of Pseudomonas aeruginosa.

    Science.gov (United States)

    Walther-Rasmussen, J; Johnsen, A H; Høiby, N

    1999-07-01

    AmpC beta-lactamases from strains of Pseudomonas aeruginosa have previously been shown to be heterogeneous with respect to their isoelectric point (pI). In order to elucidate the origin of this heterogeneity enzymes were isolated from a clinical isolate of a multiresistant P. aeruginosa strain and biochemically characterized. The purification was accomplished in four chromatographic steps comprising dye-affinity, size-exclusion, hydrophobic interaction chromatography, and chromatofocusing; this resulted in five forms with pI values of 9.1, 8.7, 8.3, 8.2, and 7.6. When analysed by SDS/PAGE and agarose IEF each separated beta-lactamase appeared to be both size- and charge-homogeneous. The specific activities of the variants were very similar. MS of each isolated beta-lactamase form showed minor differences in molecular mass (range 40.0-40.8 kDa). MS of the beta-lactamase with a pI of 8.2 demonstrated the presence of two subforms. The N-terminal sequences of three of the beta-lactamases were identical to the published sequence [Lodge, J.M. , Minchin, S.D., Piddock, L.J.V. & Busby, J.W. (1990) Biochem. J. 272, 627-631], while two variants were truncated by two amino-acid residues, one of which was acidic. The previously published sequence contains an alanine as the ultimate residue, but two of the beta-lactamases showed a substitution of Ala371 for arginine, whereas in the remaining forms C-terminal truncations by one and three residues were found. Our results indicate that the P. aeruginosa strain does not harbour multiple copies of the ampC gene, but rather that the five beta-lactamase isoforms are products of a single structural gene. The combinations of the identified N- and/or C-terminal truncations explained the multiple pI values of the beta-lactamase isoforms. PMID:10406957

  19. Regulatory components in Citrobacter freundii ampC beta-lactamase induction.

    OpenAIRE

    Lindberg, F; Westman, L; Normark, S

    1985-01-01

    Citrobacter freundii encodes an inducible chromosomal beta-lactamase similar to the constitutively expressed ampC beta-lactamase of Escherichia coli. In the latter species the ampC gene is located next to the fumarate reductase (frd) operon, whereas in C. freundii the ampC gene is known to be separated from frd by 1100 base pairs. This intervening DNA segment carries a gene, ampR, coding for a 31-kilodalton polypeptide. The cloned C. freundii OS60 ampC gene is inducible by beta-lactam antibio...

  20. Identification of group specific motifs in Beta-lactamase family of proteins

    Directory of Open Access Journals (Sweden)

    Saxena Akansha

    2009-12-01

    Full Text Available Abstract Background Beta-lactamases are one of the most serious threats to public health. In order to combat this threat we need to study the molecular and functional diversity of these enzymes and identify signatures specific to these enzymes. These signatures will enable us to develop inhibitors and diagnostic probes specific to lactamases. The existing classification of beta-lactamases was developed nearly 30 years ago when few lactamases were available. DLact database contain more than 2000 beta-lactamase, which can be used to study the molecular diversity and to identify signatures specific to this family. Methods A set of 2020 beta-lactamase proteins available in the DLact database http://59.160.102.202/DLact were classified using graph-based clustering of Best Bi-Directional Hits. Non-redundant (> 90 percent identical protein sequences from each group were aligned using T-Coffee and annotated using information available in literature. Motifs specific to each group were predicted using PRATT program. Results The graph-based classification of beta-lactamase proteins resulted in the formation of six groups (Four major groups containing 191, 726, 774 and 73 proteins while two minor groups containing 50 and 8 proteins. Based on the information available in literature, we found that each of the four major groups correspond to the four classes proposed by Ambler. The two minor groups were novel and do not contain molecular signatures of beta-lactamase proteins reported in literature. The group-specific motifs showed high sensitivity (> 70% and very high specificity (> 90%. The motifs from three groups (corresponding to class A, C and D had a high level of conservation at DNA as well as protein level whereas the motifs from the fourth group (corresponding to class B showed conservation at only protein level. Conclusion The graph-based classification of beta-lactamase proteins corresponds with the classification proposed by Ambler, thus there is

  1. Characterization of SFO-1, a plasmid-mediated inducible class A beta-lactamase from Enterobacter cloacae.

    Science.gov (United States)

    Matsumoto, Y; Inoue, M

    1999-02-01

    Enterobacter cloacae 8009 produced an inducible class A beta-lactamase which hydrolyzed cefotaxime efficiently. It also hydrolyzed other beta-lactams except cephamycins and carbapenems. The activity was inhibited by clavulanic acid and imipenem. The bla gene was transferable to Escherichia coli by electroporation of plasmid DNA. The molecular mass of the beta-lactamase was 29 kDa and its pI was 7.3. All of these phenotypic characteristics of the enzyme except for inducible production resemble those of some extended-spectrum class A beta-lactamases like FEC-1. The gene encoding this beta-lactamase was cloned and sequenced. The deduced amino acid sequence of the beta-lactamase was homologous to the AmpA sequences of the Serratia fonticola chromosomal enzyme (96%), MEN-1 (78%), Klebsiella oxytoca chromosomal enzymes (77%), TOHO-1 (75%), and FEC-1 (72%). The conserved sequences of class A beta-lactamases, including the S-X(T)-X(S)-K motif, in the active site were all conserved in this enzyme. On the basis of the high degree of homology to the beta-lactamase of S. fonticola, the enzyme was named SFO-1. The ampR gene was located upstream of the ampA gene, and the AmpR sequence of SFO-1 had homology with the AmpR sequences of the chromosomal beta-lactamases from Citrobacter diversus (80%), Proteus vulgaris (68%), and Pseudomonas aeruginosa (60%). SFO-1 was also inducible in E. coli. However, a transformant harboring plasmid without intact ampR produced a small amount of beta-lactamase constitutively, suggesting that AmpR works as an activator of ampA of SFO-1. This is the first report from Japan describing an inducible plasmid-mediated class A beta-lactamase in gram-negative bacteria. PMID:9925524

  2. Beta-Lactam-beta-lactamase-inhibitor combinations are active in experimental endocarditis caused by beta-lactamase-producing oxacillin-resistant staphylococci.

    OpenAIRE

    Hirano, L; Bayer, A S

    1991-01-01

    Optimal therapeutic strategies for serious infections caused by borderline and heterotypic oxacillin-resistant Staphylococcus aureus (BORSA and ORSA) strains have not been fully characterized. Recent evidence suggests that the dominant penicillin-binding protein of ORSA strains (PBP 2a) shows good affinity for ampicillin and that these strains commonly produce beta-lactamase. Therefore, we compared the in vivo efficacy of the combination of ampicillin plus sulbactam with that of vancomycin ag...

  3. Activity of beta-lactam beta-lactamase inhibitor combinations against extended spectrum beta-lactamase producing enterobacteriaceae in urinary isolates

    International Nuclear Information System (INIS)

    Objective: To determine the susceptibility pattern of beta-lactam beta-lactamase inhibitor combinations against extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae in urinary isolates. Study Design: Observational study. Place and Duration of Study: Ziauddin University Hospital, Karachi, from February to October 2008. Methodology: A total of 190 consecutive non-duplicate isolates of ESBL producing Enterobacteriaceae from urine samples of in-patients were included in the study. Urinary samples from out-patients, repeat samples and non-ESBL producing isolates were excluded. Detection of ESBL was carried out by double disk diffusion technique. Antimicrobial susceptibility testing was performed using modified Kirby Bauer's disk diffusion method according to CLSI guidelines. Statistical analysis was performed by SPSS version 10. Results: Of the 190 ESBL isolates tested, 88 cases (46.31%) were sensitive and 6 cases (3.15%) were resistant to all three combinations, the rest 96 cases (50.52%) were resistant to at least one of the combinations. Susceptibility pattern of cefoperazone/sulbactam, piperacillin/tazobactam, and amoxicillin/clavulanic acid was 95.26, 92.10, and 44.31 percent respectively. Conclusion: Cefoperazone/sulbactam exhibited the best activity against ESBL producing Enterobacteriaceae followed by piperacillin/tazobactam. Hospital antibiotic policies should be reviewed periodically to reduce the usage of extended spectrum cephalosporins and replace them with beta-lactam beta-lactamase inhibitor combinations agent for treating urinary tract infections. (author)

  4. Beta-Lactamase Producing Escherichia coli Isolates in Imported and Locally Produced Chicken Meat from Ghana

    DEFF Research Database (Denmark)

    Rasmussen, Mette Marie; Opintan, Japheth A; Frimodt-Møller, Niels; Styrishave, Bjarne

    2015-01-01

    by A and D, with similar distribution among the isolates from meat of locally reared chickens and imported chickens. Beta-lactamase producing genotype blaCTX-M-15 (50%; 10/20) was the most frequently drug resistant gene detected. More BLP E. coli isolates were found in imported chicken meat compared...

  5. Microbiological Screening Is Necessary to Distinguish Carriers of Plasmid-Mediated AmpC Beta-Lactamase-Producing Enterobacteriaceae and Extended-Spectrum Beta-Lactamase (ESBL)-Producing Enterobacteriaceae because of Clinical Similarity

    OpenAIRE

    Conen, Anna; Frei, Reno; Adler, Hildegard; Dangel, Marc; Fux, Christoph A; Widmer, Andreas F

    2015-01-01

    Objectives Plasmid-mediated AmpC beta-lactamase-producing (pAmpC) Enterobacteriaceae are increasing worldwide, difficult to identify and often confounded with extended-spectrum beta-lactamase (ESBL) producers. The low prevalence precludes routine universal admission screening. Therefore, we evaluated potential risk factors for carriage of pAmpC-producing Enterobacteriaceae that would allow targeted screening to improve yield and reduce cost. Patients and methods We performed a case control st...

  6. [Investigation of beta-lactamase genes and clonal relationship among the extended-spectrum beta-lactamase producing nosocomial Escherichia coli isolates].

    Science.gov (United States)

    Görgeç, Sündüz; Kuzucu, Çiğdem; Otlu, Barış; Yetkin, Funda; Ersoy, Yasemin

    2015-01-01

    Extended-spectrum beta-lactamase (ESBL) producing microorganisms currently cause a major problem. Among theseCTX-M beta-lactamase producing Escherichia coli has also disseminated worldwide as an important cause of both nosocomial and community-acquired infections. The aims of this study were to determine the prevalence of the beta-lactamase genes, antibiotic susceptibilities and clonal relationships of ESBL-producing nosocomial E.coli isolates. A total of 76 ESBL-producing E.coli strains isolated from urine (n= 26), blood (n= 25) and wound (n= 25) specimens of hospitalized patients identified as nosocomial infection agents according to the CDC criteria between June 2010-June 2011 were included in the study. Antibiotic susceptibilities of the isolates were detected by Kirby-Bauer disc diffusion method according to CLSI recommendations. ESBL production was tested by double disc diffusion method, and cefotaxime/cefotaxime-clavulanic acid E-test strips (AB Biodisk, Sweden) were used for indeterminate results. Presence of TEM, SHV, CTX-M, OXA-2 group, 0XA-10 group, PER, VEB and GES beta-lactamase genes were investigated by polymerase chain reaction (PCR) using specific primers. Pulsed-field gel electrophoresis (PFGE) method was used for the detection of clonal relationships among the strains. Most of the ESBL-producing E.coli strains were isolated from samples of inpatients in intensive care (35%), internal medicine (16%) and general surgery (13%) units. All of the 76 strains were found susceptible to imipenem, meropenem and amikacin; however all were resistant to cefotaxime and ceftriaxone. The susceptibility rates of the isolates to cefoxitin, ertapenem, cefoperazone/sulbactam, piperacillin-tazobactam, gentamicin, ciprofloxacin, cefepime, amoxicillin-clavulanic acid, aztreonam and ceftazidime were 96%, 83%, 63%, 61%, 50%, 41%, 25%, 21%, 20% and 18%, respectively. Among E.coli isolates, the frequency of CTX-M, TEM, OXA-2 group, PER, SHV and OXA-10 group beta-lactamase

  7. The revolving door between hospital and community: extended-spectrum beta-lactamase-producing Escherichia coli in Dublin.

    LENUS (Irish Health Repository)

    Burke, L

    2012-07-01

    Escherichia coli that produce extended-spectrum beta-lactamases (ESBLs) are an increasing cause of healthcare-associated infection, and community healthcare facilities may be a reservoir for important epidemic clones.

  8. Secretion of TEM beta-lactamase with signal sequences isolated from the chromosome of Lactococcus lactis subsp. lactis.

    OpenAIRE

    Sibakov, M; Koivula, T; von Wright, A.; Palva, I

    1991-01-01

    With TEM beta-lactamase as a reporter gene, a set of expression-secretion-promoting fragments were isolated from the chromosome of Lactococcus lactis subsp. lactis. The fact that only translocated beta-lactamase renders cells resistant to ampicillin allowed direct ampicillin selection with an Escherichia coli vector (pKTH33). The clones showing the greatest ampicillin resistance were subcloned onto a replicon capable of replication in lactic acid bacteria (pVS2), and the nucleotide sequences ...

  9. Multiresistant extended-spectrum beta-lactamase-producing Enterobacteriaceae from humans, companion animals and horses in central Hesse, Germany

    OpenAIRE

    Schmiedel, Judith; Falgenhauer, Linda; Domann, Eugen; Bauerfeind, Rolf; Prenger-Berninghoff, Ellen; Imirzalioglu, Can; Chakraborty, Trinad

    2014-01-01

    BACKGROUND:Multiresistant Gram-negative bacteria producing extended-spectrum beta-lactamases (ESBLs) are an emerging problem in human and veterinary medicine. This study focused on comparative molecular characterization of beta-lactamase and ESBL-producing Enterobacteriaceae isolates from central Hesse in Germany. Isolates originated from humans, companion animals (dogs and cats) and horses. RESULTS:In this study 153 (83.6%) of the human isolates (n=183) and 163 (91.6%) of the animal iso...

  10. Endemic carbapenem-resistant Pseudomonas aeruginosa with acquired metallo-beta-lactamase determinants in European hospital.

    Science.gov (United States)

    Lagatolla, Cristina; Tonin, Enrico A; Monti-Bragadin, Carlo; Dolzani, Lucilla; Gombac, Francesca; Bearzi, Claudia; Edalucci, Elisabetta; Gionechetti, Fabrizia; Rossolini, Gian Maria

    2004-03-01

    Acquired metallo-beta-lactamases (MBLs) can confer broad-spectrum beta-lactam resistance (including carbapenems) not reversible by conventional beta-lactamase inhibitors and are emerging resistance determinants of remarkable clinical importance. In 2001, multidrug-resistant Pseudomonas aeruginosa carrying bla(VIM) MBL genes were found to be widespread (approximately 20% of all P. aeruginosa isolates and 70% of the carbapenem-resistant isolates) at Trieste University Hospital. Clonal diversity and heterogeneity of resistance determinants (either bla(VIM-1)-like or bla(VIM-2)-like) were detected among MBL producers. This evidence is the first that acquired MBLs can rapidly emerge and establish a condition of endemicity in certain epidemiologic settings. PMID:15109432

  11. Resistance of Xanthomonas maltophilia to antibiotics and the effect of beta-lactamase inhibitors.

    Science.gov (United States)

    Neu, H C; Saha, G; Chin, N X

    1989-01-01

    We examined the susceptibility of 50 isolates of Xanthomonas maltophilia and the effect of beta-lactamase inhibitors upon the susceptibility. The majority of isolates were resistant to azlocillin, piperacillin, mezlocillin, ticarcillin, cefotaxime, ceftizoxime, ceftriaxone, cefoperazone, and ceftazidime. All isolates were resistant to imipenem, CGP 31608, aztreonam, and carumonam. Although disk susceptibility tests showed that the combination of clavulanate with ticarcillin inhibited many isolates, at a ratio of 1:20 few isolates were susceptible to the combination. Addition of clavulanate to aztreonam and to imipenem failed to make organisms susceptible. Sulbactam combined with cefoperazone made some organisms susceptible, but ampicillin-sulbactam was ineffective, whereas tazobactam combined with piperacillin at a ratio of 1:4 made half the isolates have MICs of 32 micrograms/ml or less. The beta-lactamases from the isolates hydrolyzed all of the beta-lactams. PMID:2791491

  12. Lactam hydrolysis catalyzed by mononuclear metallo-beta-lactamases: A density functional study

    DEFF Research Database (Denmark)

    Hemmingsen, Lars Bo Stegeager; Olsen, L.; Antony, J.; Ryde, U.; Adolph, H.-W.

    2003-01-01

    Two central steps in the hydrolysis of lactam antibiotics catalyzed by mononuclear metallo-beta-lactamases, formation of the tetrahedral intermediate and its breakdown by proton transfer, are studied for model systems using the density functional B3LYP method. Metallo-beta-lactamases have two metal......-limiting step. The barrier for the breakdown of the intermediate is low, 0-10 kcal/mol, if it is assisted by a water molecule or by a Cys or Asp model. Thus, the results indicate that proton transfer is not rate-limiting, and that any of the residues from the second metal site may function as proton shuttle...... model with Asp as a proton shuttle, attack of the zinc-bond hydroxide ion seems to be concerted with the proton transfer. We have also studied the effect of replacing one of the histidine ligands by an asparagine or glutamine residue, giving a zinc site representative of other subclasses of metallo...

  13. The role of beta-lactamase-producing-bacteria in mixed infections

    Directory of Open Access Journals (Sweden)

    Brook Itzhak

    2009-12-01

    Full Text Available Abstract Beta-lactamase-producing bacteria (BLPB can play an important role in polymicrobial infections. They can have a direct pathogenic impact in causing the infection as well as an indirect effect through their ability to produce the enzyme beta-lactamase. BLPB may not only survive penicillin therapy but can also, as was demonstrated in in vitro and in vivo studies, protect other penicillin-susceptible bacteria from penicillin by releasing the free enzyme into their environment. This phenomenon occurs in upper respiratory tract, skin, soft tissue, surgical and other infections. The clinical, in vitro, and in vivo evidence supporting the role of these organisms in the increased failure rate of penicillin in eradication of these infections and the implication of that increased rate on the management of infections is discussed.

  14. Phenotypic and Molecular Characterization of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Bangladesh

    OpenAIRE

    Lina, Taslima T.; Khajanchi, Bijay K.; Azmi, Ishrat J.; Mohammad Aminul Islam; Belal Mahmood; Mahmuda Akter; Atanu Banik; Rumana Alim; Armando Navarro; Gabriel Perez; Alejandro Cravioto; Talukder, Kaisar A

    2014-01-01

    BACKGROUND: Resistance to cephalosporins in Enterobacteriaceae is mainly due to the production of extended-spectrum beta-lactamase (ESBL). Little is known about ESBL-producing bacteria in Bangladesh. Therefore, the study presents results of phenotypic and molecular characterization of ESBL-producing Escherichia coli from hospitals in Bangladesh. METHODS: A total of 339 E. coli isolated from patients with urinary tract and wound infections attending three different medical hospitals in urban a...

  15. Extended spectrum beta-lactamases in Escherichia coli from municipal wastewater

    OpenAIRE

    Tatiana Čornejová; Jan Venglovsky; Gabriela Gregova; Marta Kmetova; Vladimir Kmet

    2015-01-01

    Introduction and objective. Over the past decades, awareness of the environmental load of resistant organisms has increased. The presented paper focuses on antibiotic resistance and detection of resistance genes in environmental [i]E. coli[/i] and on the evaluation of biofilm formation in ESBLs (extended spectrum beta-lactamase) producing [i]E. coli[/i] isolated from an urban wastewater treatment plant. Materials and method. Wastewater samples and artificially added polystyrene pellets ...

  16. Surveillance and Detection of Inhibitor-Resistant Beta-Lactamases in Clinical Isolates of Escherichia coli

    Directory of Open Access Journals (Sweden)

    Carl Urban

    2007-06-01

    Full Text Available In recent years, resistance to beta-lactam antibiotics, such as the widely-used cephalosporins and penicillins, has become a major challenge for disease therapy, particularly in common hospital-acquired infections. In the search for the mechanisms behind this increasingly prevalent form of resistance, microbiologists have identified a new type of beta-lactamase enzyme, called inhibitor-resistant TEMs (IRTs, which can withstand the effects of beta-lactamase inhibitor compounds, further reducing the arsenal of drugs available to physicians facing resistant bacteria. In this study, we examined the enzymatic and genetic basis of Escherichia coli isolates demonstrating such resistance to beta-lactam/beta-lactamase inhibitor combinations. Susceptibility trials played a major role in composing the experimental cohort for this project (n=50; each isolate was thoroughly tested to ensure that it was resistant to ampicillin-sulbactam, an inhibitor combination, but susceptible to the third-generation cephalosporin ceftazidime. Subsequently, a number of samples were subjected to assay by pulsed-field gel electrophoresis (n=18 and polymerase chain reaction (n=3 so that their genetic composition and relatedness might be known. In particular, the presence of genes coding for TEM-type beta-lactamases was investigated for each of the 3 isolates sequenced. Even though it was anticipated that the isolates would possess resistance to inhibitor combinations due to an IRT gene, this was not found to be the case. Instead, the mechanism of resistance turned out to be over-expression of a gene coding for a normal TEM enzyme. The results of these experiments have implications for ensuring successful therapy of bacterial infections and for preventing the spread of antimicrobial resistance.

  17. Dissemination of extended-spectrum beta-lactamase producing enterobacteriaceae in children

    OpenAIRE

    Lima, S.B.; Ferreira, H.N.

    2013-01-01

    ABSTRACT Extended-spectrum beta-lactamase producing Enterobacteriaceae (EESBL), have been considered important nosocomial pathogens during the last decades. Nowadays community dissemination of this resistance threat is a reality, namely, in particular niches, as old people care settings. Fecal colonization of healthy people is a reality that might compromise effective infection control in acute care hospitals and long term care facilities and in that way, screening of ...

  18. Metallo-beta-lactamases as emerging resistance determinants in Gram-negative pathogens: open issues

    OpenAIRE

    Cornaglia, G; Akova, M; Amicosante, G; Canton, R.; Cauda, R.; J.D. Docquier; Edelstein, M.; Frère, Jean-Marie; Fuzi, M.; Galleni, Moreno; Giamarellou, H; Gniadkowski, M.; Koncan, R.; Libisch, B.; F. Luzzaro

    2007-01-01

    The rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is a matter of particular concern worldwide and primarily in Europe, one of first continents where the emergence of acquired MBLs has been reported and possibly the geographical area where the increasing diversity of these enzymes and the number of bacterial species affected are most impressive. This spread has not been paralleled by accuracy/standardisation of detection methods, completeness of ep...

  19. Burn Patients Infected With Metallo-Beta-Lactamase-Producing Pseudomonas aeruginosa: Multidrug-Resistant Strains

    OpenAIRE

    Anvarinejad, Mojtaba; Japoni, Aziz; Rafaatpour, Noroddin; Mardaneh, Jalal; Abbasi, Pejman; Amin Shahidi, Maneli; Dehyadegari, Mohammad Ali; Alipour, Ebrahim

    2014-01-01

    Background: Metallo-beta-lactamase (MBL) producing Pseudomonas aeruginosa in the burn patients is a leading cause of morbidity and mortality and remains a serious health concern among the clinicians. Objectives: The aim of this study was to detect MBL-producing P. aeruginosa in burn patients and determine multidrug-resistant (MDR) strains, and respective resistance patterns. Patients and Methods: In this cross-sectional study, 270 strains of P. aeruginosa were isolated from the burn patients ...

  20. Novel plasmid-mediated beta-lactamase from Escherichia coli that inactivates oxyimino-cephalosporins.

    OpenAIRE

    Matsumoto, Y; Ikeda, F; Kamimura, T.; Yokota, Y.; Mine, Y

    1988-01-01

    A highly cephem-resistant Escherichia coli strain, FP1546, isolated from the fecal flora of laboratory dogs previously administered beta-lactam antibiotics was found to produce a beta-lactamase, FEC-1, of 48-kilodalton size and pI 8.2. FEC-1 hydrolyzed cefuroxime, cefotaxime, cefmenoxime, and ceftriaxone, as well as the enzymatically less-stable antibiotics cephaloridine, cefotiam, and cefpiramide. Of the oxyimino-cephalosporins, ceftizoxime was fairly stable to FEC-1. FEC-1 differed notably ...

  1. Extended spectrum beta lactamase producing Proteus penneri: A rare missed pathogen?

    OpenAIRE

    Anita Pandey; Himani Verma; Ashish K Asthana; Molly Madan

    2014-01-01

    Indole negative Proteus species are invariably incorrectly identified as Proteus mirabilis, often missing out isolates of Proteus penneri. We report a case of extended spectrum beta lactamase producing and multidrug-resistant P. penneri isolated from pus from pressure sore of a patient of road traffic accident. Correct and rapid isolation and identification of such resistant pathogen are important as they are significant nosocomial threat.

  2. Antibacterial Activity of Some Plant Extracts Against Extended- Spectrum Beta-Lactamase Producing Escherichia coli Isolates

    OpenAIRE

    Saeidi, Saeide; Amini Boroujeni, Negar; Ahmadi, Hassan; Hassanshahian, Mehdi

    2015-01-01

    Background: The extended-spectrum beta-lactamase (ESBL) -producing Escherichia coli isolates make many serious infections, especially urinary tract infections. Objectives: The purpose of this study was to determine the antibacterial activities of some natural plant extracts against ESBL-producing E. coli isolates, which harbor the TEM gene in urine samples of the patients who have urinary tract infections. Materials and Methods: Evaluation has to be exactly determined for both methods of disk...

  3. AmpC-BETA Lactamases among Enterobacteriaceae Isolated at a Tertiary Hospital, South Western Uganda

    OpenAIRE

    Nakaye, Martha; Bwanga, Freddie; Itabangi, Herbert; Stanley, Iramiot J.; Bashir, Mwambi; Bazira, Joel

    2014-01-01

    Aim To characterize AmpC-beta lactamases among Enterobacteriaceae isolates from clinical samples at Mbarara Regional Referral Hospital. Study Design Laboratory-based descriptive cross-sectional study Place and Duration of Study Microbiology Department, Mbarara Regional Referral Hospital and MBN clinical Laboratories, between May to September 2013. Methodology This study included 293 Enterobacteriaceae isolates recovered from clinical specimens that included blood, urine, stool and aspirates. ...

  4. Cephamycin inactivation due to enzymatic hydrolysis by beta-lactamase from Bacteroides fragilis.

    OpenAIRE

    Yotsuji, A; Minami, S; Kakizawa, H; Yasuda, T; Takai, A.; Saikawa, I; Inoue, M.; Mitsuhashi, S

    1985-01-01

    The susceptibility of 53 clinical isolates of Bacteroides fragilis to cephamycins was examined. Judging from the MICs for 50% of the strains tested, moxalactam was the most active, however, judging from the MICs for 90% of the strains tested, cefbuperazone was more effective than moxalactam. A correlation was observed between in vitro activity of benzylpenicillin and cephaloridine and beta-lactamase production. Inactivation due to enzymatic hydrolysis of cephamycins over a short time was not ...

  5. Studies on structure-based sequence alignment and phylogenies of beta-lactamases.

    Science.gov (United States)

    Salahuddin, Parveen; Khan, Asad U

    2014-01-01

    The β-lactamases enzymes cleave the amide bond in β-lactam ring, rendering β-lactam antibiotics harmless to bacteria. In this communication we have studied structure-function relationship and phylogenies of class A, B and D beta-lactamases using structure-based sequence alignment and phylip programs respectively. The data of structure-based sequence alignment suggests that in different isolates of TEM-1, mutations did not occur at or near sequence motifs. Since deletions are reported to be lethal to structure and function of enzyme. Therefore, in these variants antibiotic hydrolysis profile and specificity will be affected. The alignment data of class A enzyme SHV-1, CTX-M-15, class D enzyme, OXA-10, and class B enzyme VIM-2 and SIM-1 show sequence motifs along with other part of polypeptide are essentially conserved. These results imply that conformations of betalactamases are close to native state and possess normal hydrolytic activities towards beta-lactam antibiotics. However, class B enzyme such as IMP-1 and NDM-1 are less conserved than other class A and D studied here because mutation and deletions occurred at critically important region such as active site. Therefore, the structure of these beta-lactamases will be altered and antibiotic hydrolysis profile will be affected. Phylogenetic studies suggest that class A and D beta-lactamases including TOHO-1 and OXA-10 respectively evolved by horizontal gene transfer (HGT) whereas other member of class A such as TEM-1 evolved by gene duplication mechanism. Taken together, these studies justify structure-function relationship of beta-lactamases and phylogenetic studies suggest these enzymes evolved by different mechanisms. PMID:24966539

  6. Detection of Beta-Lactamase and Extended-Spectrum Beta-Lactamase of Pathogens Isolated from Pig and Chicken and Their Antibiotic Susceptibility Test

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    The antibacterial activity of beta-lactam antibiotics or their combinations with inhibitor sulbactum against non-lactamaseproducing strains, lactamase-producing and ESBLs-producing isolates was evaluated with twofold dilution method after pathogens isolated from pigs and chickens were detected, respectively, for beta-lactamase and extended-spectrum betalactamases (ESBLs). The results revealed that most of 43 clinically isolated strains could produce beta-lactamase and 3strains of shigella isolated from chicken samples produced ESBLs. All of 30 lactamase-producing strains isolated and only one of 16 non-lactamase-producing strains were resistant to amoxicillin and ampicillin. MICs of ampicillin against lactamaseproducing isolates decreased 10-40 and 10-20 times respectively, when it was conbined with sulbactam at ration of 1:2 and 1:4. All clinical isolates were susceptible to third-generation cephalosporins. The MICs of third-generation cephalosporins against lactamase-producing isolates did not change when they were conbined with sulbactam. MICs of ceftiofur and ceftriaxone against ESBLs-producing isolates decreased 2-4 times when they were conbined with sulbactam.

  7. The Deacylation Mechanism of AmpC [beta]-Lactamase at Ultrahigh Resolution

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Yu; Minasov, George; Roth, Tomer A.; Prati, Fabio; Shoichet, Brian K. (Degli); (NWU); (UCSF)

    2010-03-05

    {beta}-Lactamases confer bacterial resistance to {beta}-lactam antibiotics, such as penicillins. The characteristic class C {beta}-lactamase AmpC catalyzes the reaction with several key residues including Ser64, Tyr150, and Lys67. Here, we describe a 1.07 {angstrom} X-ray crystallographic structure of AmpC {beta}-lactamase in complex with a boronic acid deacylation transition-state analogue. The high quality of the electron density map allows the determination of many proton positions. The proton on the Tyr150 hydroxyl group is clearly visible and is donated to the boronic oxygen mimicking the deacylation water. Meanwhile, Lys67 hydrogen bonds with Ser64O{gamma}, Asn152O{delta}1, and the backbone oxygen of Ala220. This suggests that this residue is positively charged and has relinquished the hydrogen bond with Tyr150 observed in acyl-enzyme complex structures. Together with previous biochemical and NMR studies, these observations indicate that Tyr150 is protonated throughout the reaction coordinate, disfavoring mechanisms that involve a stable tyrosinate as the general base for deacylation. Rather, the hydroxyl of Tyr150 appears to be well positioned to electrostatically stabilize the negative charge buildup in the tetrahedral high-energy intermediate. This structure, in itself, appears consistent with a mechanism involving either Tyr150 acting as a transient catalytic base in conjunction with a neutral Lys67 or the lactam nitrogen as the general base. Whereas mutagenesis studies suggest that Lys67 may be replaced by an arginine, disfavoring the conjugate base mechanism, distinguishing between these two hypotheses may ultimately depend on direct determination of the pKa of Lys67 along the reaction coordinate.

  8. Detection of Amp C genes encoding for beta-lactamases in Escherichia coli and Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    M Shanthi

    2012-01-01

    Full Text Available Purpose : Amp C beta-lactamase are Ambler class C enzymes that confer resistance to extended spectrum cephalosporins and are not inhibited by beta-lactamase inhibitors. Their detection is crucial, since the phenotypic tests are not standardised leading to ambiguity in interpretation of results. This study was done to detect the types of Amp C prevalent in Escherichia coli and Klebsiella pneumoniae by multiplex polymerase chain reaction (PCR. Materials and Methods : Seventy-seven consecutive cefoxitin resistant clinical isolates of E. coli (n = 25 and K. pneumoniae (n = 52 were included in the study. Antibiotic susceptibility testing to various classes of antibiotics was performed by disc diffusion using Clinical Laboratory Standards Institute (CLSI guidelines. Minimum inhibitory concentration (MIC to cefoxitin, imipenem and meropenem were determined by broth microdilution method. Isolates were screened for production of Extended Spectrum Beta-Lactamase (ESBL. Multiplex PCR was performed for the detection of Amp C genes after phenotypic testing (Hodge test and inhibitor based test. Results : Cefoxitin Hodge test was positive in 40 isolates which included 20 E. coli and 20 K. pneumoniae. There was zone enhancement with boronic acid in 55 isolates, of which 36 were K. pneumoniae and 19 were E. coli. Multiplex PCR detected Amp C in 11/25 E. coli and 12/52 K. pneumoniae isolates. The Amp C genes detected were CIT (Amp C origin - Citrobacter freundii, DHA (Dhahran Hospital, Saudi Arabia, ACC (Ambler class C, EBC (Amp C origin - Enterobacter cloacae groups. ESBL was co-produced in 54 isolates. Conclusions : Amp C was detected in 29.87% of the study isolates. Majority of them co-produced ESBL. The most common Amp C was the CIT family. Screen tests for cefoxitin resistance may be falsely positive due to production of carbapenamases.

  9. Purification and biochemical characterization of the VIM-1 metallo-beta-lactamase.

    Science.gov (United States)

    Franceschini, N; Caravelli, B; Docquier, J D; Galleni, M; Frère, J M; Amicosante, G; Rossolini, G M

    2000-11-01

    VIM-1 is a new group 3 metallo-beta-lactamase recently detected in carbapenem-resistant nosocomial isolates of Pseudomonas aeruginosa from the Mediterranean area. In this work, VIM-1 was purified from an Escherichia coli strain carrying the cloned bla(VIM-1) gene by means of an anion-exchange chromatography step followed by a gel permeation chromatography step. The purified enzyme exhibited a molecular mass of 26 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and an acidic pI of 5.1 in analytical isoelectric focusing. Amino-terminal sequencing showed that mature VIM-1 results from the removal of a 26-amino-acid signal peptide from the precursor. VIM-1 hydrolyzes a broad array of beta-lactam compounds, including penicillins, narrow- to expanded-spectrum cephalosporins, carbapenems, and mechanism-based serine-beta-lactamase inactivators. Only monobactams escape hydrolysis. The highest catalytic constant/K(m) ratios (>10(6) M(-1). s(-1)) were observed with carbenicillin, azlocillin, some cephalosporins (cephaloridine, cephalothin, cefuroxime, cefepime, and cefpirome), imipenem, and biapenem. Kinetic parameters showed remarkable variability with different beta-lactams and also within the various penam, cephem, and carbapenem compounds, resulting in no clear preference of the enzyme for any of these beta-lactam subfamilies. Significant differences were observed with some substrates between the kinetic parameters of VIM-1 and those of other metallo-beta-lactamases. Inactivation assays carried out with various chelating agents (EDTA, 1,10-o-phenanthroline, and pyridine-2,6-dicarboxylic acid) indicated that formation of a ternary enzyme-metal-chelator complex precedes metal removal from the zinc center of the protein and revealed notable differences in the inactivation parameters of VIM-1 with different agents. PMID:11036013

  10. Prevalence of Extended –Spectrum-Beta-Lactamase-Producing Klebsiella Pneumonia Isolates from Clinical Samples

    Directory of Open Access Journals (Sweden)

    Alizade, H. (MSc

    2014-06-01

    Full Text Available Background and Objective: Klebsiella pneumonia (K.pneumonia is one of the common causes of nosocomial infections. The aim of this research was to determine the prevalence of beta-lactamase genes and phenotypic confirmation of extended–spectrum-beta-lactamase (ESBL producing K.pneumonia isolates from clinical samples. Material and Methods: In this study, 122 K.pneumonia were isolated from clinical specimens of Khoramabad city and were confirmed by standard bacteriological tests. The presence of ESBL enzymes was detected by combined disk diffusion method. PCR assay with specific primers was used to determine blaSHV, blaTEM, blaCTX-15 and blaCTX-M genes in the confirmed isolates. Results: of 122 K.pneumonia isolates, 78 (64.18% were positive for ESBL, using disk diffusion method. According to antibiogram results, 10.65% of isolates were resistant to cefotaxime, 3.27% to ceftazidime and 68.03% to both antibiotics. Ninety isolates (64.18% considered as ESBLs isolates, at the same time, with being resistant to cefotaxime and ceftazidime were also sensitive to cefotaxime-clavulanic acid and ceftazidime-clavulanic acid. In PCR assays, blaCTX-15, blaSHV, blaCTX-M and blaTEM genes were detected in 78.68%, 40.16%, 26.22% and 22.13% of isolates, respectively. Ten resistant patterns of genes were detected. Conclusion: The significance percentage of antibiotic resistant genes of K.pneumonia isolates from clinical samples in Khoramabad city had ESBLs genes; CTX-M category was the most prevalent encoding genes of these enzymes. Keywords: Klebsiella Pneumonia, Extended-Spectrum Beta-Lactamase, Antibiotic Resistance

  11. Detection of genes mediating beta-lactamase production in isolates of enterobacteria recovered from wild pets in Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Sabry A. Hassan

    2015-12-01

    Full Text Available Aim: To determine the genetic basis and types of beta-lactamase encountered among enterobacterial isolates of wild pets from the animal exhibit. Materials and Methods: A total of 17 beta-lactamase-producing enterobacteria recovered from fecal samples of wild pet animals were analyzed for a selected beta-lactamase gene by polymerase chain reaction. Results: Molecular analysis identified one or more β-lactamase-encoding genes in 14 enterobacterial isolates as a single or gene combination. The most frequent extended-spectrum β-lactamases types were transmission electron microscopy and CTX-M, and the most common AmpC enzymes were CMY-2 and DHA types. Conclusions: The study is the first in Saudi Arabia, have established the presence of β-lactamase-encoding genes in the fecal isolates of wild pets.

  12. Prevalence of beta-lactamases among ampicillin-resistant Escherichia coli and Salmonella isolated from food animals in Denmark

    DEFF Research Database (Denmark)

    Olesen, Inger; Hasman, Henrik; Aarestrup, Frank Møller

    2004-01-01

    variants of bla(TEM-1), of which bla(TEM-1b) was the most frequently detected (80 E. coli and 47 Salmonella), followed by bla(TEM-1a) (eight E. coli, one Salmonella) and bla(TEM-1c) (seven E. coli). A few isolates were found to express OXA, TEM-30, or PSE beta-lactamases. Mutations in the ampC promoter...... leading to increased production of the AmpC beta-lactamase were demonstrated in 11 cefoxitin-resistant or intermediate E. coli isolates. Nine of these isolates did not contain any bla(TEM) genes, whereas the remaining two did. No genes encoding SHV or extended-spectrum beta-lactamases were detected. Two...

  13. The effects on the expression of. beta. -lactamase by targeted insertion of a Kirsten murine leukemia virus variant into the coding region of the gene

    Energy Technology Data Exchange (ETDEWEB)

    Dias-Ferrao, V.P.T.

    1988-01-01

    The product of this plasmid gene protects bacteria from the antibiotic, ampicillin. When the Kirsten murine leukemia virus variant DNA (MuLV-K-Vd) was inserted into the Pst 1 site of the {beta}-lactamase gene, the transformed bacteria (E. coli, DH5) were resistant to ampicillin. The purpose of this study is to explain the presence of a functional {beta}-lactamase gene with additional nucleotides inserted into the coding region of the gene. The recombinant plasmid codes for a functional {beta}-lactamase. Northern blot analysis of RNA using a {sup 32}P-labelled 16{sup mer} oligonucleotide as a probe revealed the {beta}-lactamase transcript from the recombinant plasmid to be shorter than the transcript from the wild-type {beta}-lactamase gene. Also, greater levels of {beta}-lactamase mRNA were present in cells containing the recombinant plasmid compared to those containing the wild-type plasmid. Restriction enzyme mapping indicated that the 3{prime} end of MuLV-K-Vd insert contains sequences of {beta}-lactamase. Nucleic acid sequencing substantiated the hybridization data that {beta}-lactamase sequences are present in the 3{prime} end of MuLV-K-Vd. However, exact sequence homology is not evident.

  14. Antibodies against Pseudomonas aeruginosa chromosomal beta-lactamase inpatients with cystic fibrosis are markers of the development of resistance of P. aeruginosa to beta-lactams

    DEFF Research Database (Denmark)

    Ciofu, O; Giwercman, B; Walter-Rasmussen, J; Pressler, T; Pedersen, S S; Høiby, N

    1995-01-01

    Chromosomal beta-lactamase production is considered to be the most important resistance mechanism of Pseudomonas aeruginosa against beta-lactams. Recently we have detected serum and sputum antibodies against P. aeruginosa chromosomal beta-lactamase (a beta ab), using immunoblotting techniques. In...

  15. Beta-Lactamase Producing Escherichia coli Isolates in Imported and Locally Produced Chicken Meat from Ghana.

    Directory of Open Access Journals (Sweden)

    Mette Marie Rasmussen

    Full Text Available The use of antibiotics in food animals is of public health concern, because resistant zoonotic pathogens can be transmitted to humans. Furthermore, global trade with food may rapidly spread multi-resistant pathogens between countries and even continents. The purpose of the study was to investigate whether imported chicken meat and meat from locally reared chicken are potential sources for human exposure to multi resistant Escherichia coli isolates. 188 samples from imported and locally produced chicken meat were sampled and analyzed. 153 bacteria isolates were successfully cultured and identified as E. coli using MALDI-ToF. Of these 109 isolates were from meat whereas the remaining 44 were isolated from the cloaca of locally reared live chickens. Antimicrobial susceptibility test was done on the identified E. coli isolates. Additionally, beta-lactamases production (ESBL and/or AmpC were phenotypically confirmed on all isolates showing resistance to cefpodoxime. Beta-lactamase producing (BLP E. coli meat isolates were further genotyped. Antimicrobial resistance to four antibiotic markers with highest resistance was detected more frequently in isolates from local chickens compared to imported chickens (tetracycline 88.9% vs. 57.5%, sulphonamide 75.0% vs. 46.6%, ampicillin 69.4% vs. 61.6% and trimethoprim 66.7% vs. 38.4%. Beta-lactamase production was found in 29 E. coli meat isolates, with 56.9% of them being multiple drug resistant (≥ 3. The predominant phylogroup identified was B1 followed by A and D, with similar distribution among the isolates from meat of locally reared chickens and imported chickens. Beta-lactamase producing genotype blaCTX-M-15 (50%; 10/20 was the most frequently drug resistant gene detected. More BLP E. coli isolates were found in imported chicken meat compared to locally reared chickens, demonstrating that these isolates may be spreading through food trade. In conclusion, both imported and locally produced chicken meats

  16. [First outbreak report of VIM-1 metallo-beta-lactamase producing Pseudomonas aeruginosa in Japan].

    Science.gov (United States)

    Miki, Kanji; Takegawa, Hiroshi; Etoh, Masaaki; Hayashi, Michio; Haruta, Tsunekazu; Yamane, Kunikazu; Arakawa, Yoshichika

    2010-11-01

    VIM-1 metallo-beta-lactamase (MBL) producing Pseudomonas aeruginosa was isolated from 35 Kobe City Medical Center General Hospital patients from September 2007 to July 2008. All but one were highly resistant to all beta-lactams, aminoglycoside, and fluoroquinolone, and one susceptible to amikacin. Strains negative to a disk diffusion screening test using sodium mercaptoacetate for detecting MBL numbered 35. PCR for MBL indicated all strains were positive for bla(VIWM-1). These strains were indistinguishable by pulsed-field gel electrophoresis, indicating an outbreak of infections caused by VIM-1 MBL producing Pseudomonas aeruginosa. After intervention to control contact, the outbreak was controlled. PMID:21226324

  17. Expression, purification, crystallization and preliminary X-ray analysis of Aeromonas hydrophilia metallo-[beta]-lactamase

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, N.; Toney, J.H.; Fitzgerald, P.M.D. (Merck)

    2010-07-20

    The CphA metallo-{beta}-lactamase from Aeromonas hydrophilia has been expressed, purified and crystallized by the hanging-drop vapor-diffusion method using ammonium sulfate as the precipitant. The crystals exhibit orthorhombic symmetry (P2{sub 1}2{sub 1}2), with unit-cell parameters a = 40.75, b = 42.05, c = 128.88 {angstrom}. There is one monomer in the asymmetric unit and the solvent content is estimated to be 44% by volume. A data set extending to 1.8 {angstrom} has been measured.

  18. Extended-spectrum beta-lactamase production among ampicillin-resistant Escherichia coli strains from chicken in Enugu State, Nigeria Produção de beta-lactamase de espectro expandido por cepas de Escherichia coli resistentes a ampicilina isoladas de frango em Enugu State, Nigéria

    OpenAIRE

    Chah, K.F.; Oboegbulem, S. I.

    2007-01-01

    One hundred and seventy-two ampicillin-resistant E. coli strains isolated from commercial chickens in Enugu State, Nigeria, were screened for beta-lactamase production using the broth method with nitrocefin® as the chromogenic cephalosporin to detect enzyme production. Beta-lactamase producing strains were further examined for extended-spectrum beta-lactamase (ESBL) production using the Oxoid combination discs method. One hundred and seventy (98.8%) of the 172 ampicillin-resistant E. coli str...

  19. Antibodies against beta-lactamase can improve ceftazidime treatment of lung infection with beta-lactam-resistant Pseudomonas aeruginosa in a rat model of chronic lung infection

    DEFF Research Database (Denmark)

    Ciofu, Oana; Bagge, Niels; Høiby, Niels

    2002-01-01

    To test the hypothesis that antibodies against the chromosomal beta-lactamase of Pseudomonas aeruginosa (a beta ab) might act as beta-lactamase inhibitors in patients with cystic fibrosis and chronic lung infection with P. aeruginosa, we compared in a rat model of chronic lung infection the...... efficacy of treatment with ceftazidime in beta-lactamase-immunized (group I) and non-immunized (group II) rats. Chronic lung infection was established with alginate-embedded P. aeruginosa producing high amounts of beta-lactamase in 133 Lewis rats. Prior to infection, group I (66 rats) was immunized three...... times at 2-week intervals with purified beta-lactamase in incomplete Freund's adjuvant (IFA) and group II (67 rats) received IFA. Ceftazidime treatment was initiated after challenge and continued for 10 days, after which the rats were sacrificed and the lung bacteriology and pathology were analysed. Rat...

  20. Pharmacokinetic studies and renal dehydropeptidase stability of the new beta-lactamase inhibitor BRL 42715 in animals.

    OpenAIRE

    Coleman, K.; Griffin, D. R.; Upshon, P A

    1991-01-01

    BRL 42715 is a novel, highly potent beta-lactamase inhibitor with good activity against a broad range of beta-lactamases, including the class I enzymes of Enterobacter and Citrobacter spp. (K. Coleman, D.R.J. Griffin, J.W.J. Page, and P.A. Upshon, Antimicrob. Agents Chemother. 33:1580-1587, 1989). The pharmacokinetics of BRL 42715 were studied following oral and parenteral administration in mice, rats, rabbits, beagle dogs, and cynomolgus monkeys. The elimination half-life (t1/2) of BRL 42715...

  1. Activities of beta-lactam antibiotics against Escherichia coli strains producing extended-spectrum beta-lactamases.

    OpenAIRE

    Jacoby, G A; Carreras, I

    1990-01-01

    Seven extended-spectrum beta-lactamases related to TEM and four enzymes derived from SHV-1 were transferred to a common Escherichia coli host so that the activity of a variety of beta-lactams could be tested in a uniform genetic environment. For most derivatives, penicillinase activity was 10% or less than that of strains making TEM-1, TEM-2, or SHV-1 beta-lactamase, suggesting that reduced catalytic efficiency accompanied the broader substrate spectrum. Despite this deficit, resistance to az...

  2. Susceptibility pattern of extended spectrum beta-lactamase producing isolates in various clinical specimens

    International Nuclear Information System (INIS)

    Objective: To determine the susceptibility pattern of extended spectrum beta-lactamase (ESBL) producing Gram negative isolates from various clinical specimens. Study Design: Descriptive study. Place and Duration of Study: Microbiology Department, Armed Forces Institute of Pathology, Rawalpindi, from January 2008 to January 2009. Methodology: A total of 308 ESBL producing isolates from various clinical specimens sent to AFIP for culture and sensitivity were identified using standard microbiological techniques and tested for antimicrobial susceptibility. At the same time screening for ESBL production was also done. ESBL production was confirmed by combination disc synergy method. The susceptibility pattern of isolates was then recorded in frequency percentages. Results: Out of the 308 ESBL producing isolates more than 99% were susceptible to carbapenems, 84% to tazobactam/ piperacillin, 81% to sulbactam/cefoperazone, 12% to fluoroquinolones, 13% to cotrimoxazole, 59% to amikacin and 18% to gentamicin. Among the urinary isolates 49% were susceptible to Nitrofurontoin and only 5% to Pipemidic acid. Conclusion: Antibiotic choices in case of ESBL producing isolates are limited and at present only carbapenems can be regarded as treatment of choice. As empirical agents, beta-lactam/beta lactamase inhibitor combinations should be used cautiously for serious infections. Fluoroquinolones showed very poor efficacy. Amikacin can be used alternatively in such cases. Nitrofurantoin is still a good oral agent for treating UTI. (author)

  3. Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

    Directory of Open Access Journals (Sweden)

    Ronak Bakhtiari

    2011-10-01

    Full Text Available Resistance to b-lactam antibiotics by gramnegative bacteria, especially Escherichia coli (E. coli, is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly E. coli 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 E. coli isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on E. coli 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% E. coli 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.

  4. Exposure assessment of extended-spectrum beta-lactamases/AmpC beta-lactamases-producing Escherichia coli in meat in Denmark

    Directory of Open Access Journals (Sweden)

    Luís P. Carmo

    2014-02-01

    Full Text Available Introduction: Extended-spectrum beta-lactamases (ESBL and AmpC beta-lactamases (AmpC are of concern for veterinary and public health because of their ability to cause treatment failure due to antimicrobial resistance in Enterobacteriaceae. The main objective was to assess the relative contribution (RC of different types of meat to the exposure of consumers to ESBL/AmpC and their potential importance for human infections in Denmark. Material and methods: The prevalence of each genotype of ESBL/AmpC-producing E. coli in imported and nationally produced broiler meat, pork and beef was weighted by the meat consumption patterns. Data originated from the Danish surveillance program for antibiotic use and antibiotic resistance (DANMAP from 2009 to 2011. DANMAP also provided data about human ESBL/AmpC cases in 2011, which were used to assess a possible genotype overlap. Uncertainty about the occurrence of ESBL/AmpC-producing E. coli in meat was assessed by inspecting beta distributions given the available data of the genotypes in each type of meat. Results and discussion: Broiler meat represented the largest part (83.8% of the estimated ESBL/AmpC-contaminated pool of meat compared to pork (12.5% and beef (3.7%. CMY-2 was the genotype with the highest RC to human exposure (58.3%. However, this genotype is rarely found in human infections in Denmark. Conclusion: The overlap between ESBL/AmpC genotypes in meat and human E. coli infections was limited. This suggests that meat might constitute a less important source of ESBL/AmpC exposure to humans in Denmark than previously thought – maybe because the use of cephalosporins is restricted in cattle and banned in poultry and pigs. Nonetheless, more detailed surveillance data are required to determine the contribution of meat compared to other sources, such as travelling, pets, water resources, community and hospitals in the pursuit of a full source attribution model.

  5. In-vitro activity and beta-lactamase stability of methicillin, isoxazolyl penicillins and cephalothin against coagulase-negative staphylococci

    DEFF Research Database (Denmark)

    Jarløv, J O; Rosdahl, V T; Mortensen, I;

    1988-01-01

    the isoxazolyl penicillins showed a high degree of uniformity. However more strains were resistant to cloxacillin and oxacillin than to dicloxacillin and flucloxacillin. Only a weak correlation was found between beta-lactamase production, and resistance to the six antibiotics. Methicillin was the most...

  6. Prevalence of beta-lactamases among ampicillin-resistant Escherichia coli and Salmonella isolated from food animals in Denmark

    DEFF Research Database (Denmark)

    Olesen, Inger; Hasman, Henrik; Aarestrup, Frank Møller

    2004-01-01

    The genetic background for beta-lactamase-mediated resistance to beta-lactam antibiotics was examined by PCR and sequencing in 160 ampicillin-resistant isolates (109 Escherichia coli and 51 Salmonella) obtained from healthy and diseased food animals in Denmark. Sequencing revealed three different...... variants of bla(TEM-1), of which bla(TEM-1b) was the most frequently detected (80 E. coli and 47 Salmonella), followed by bla(TEM-1a) (eight E. coli, one Salmonella) and bla(TEM-1c) (seven E. coli). A few isolates were found to express OXA, TEM-30, or PSE beta-lactamases. Mutations in the ampC promoter...... leading to increased production of the AmpC beta-lactamase were demonstrated in 11 cefoxitin-resistant or intermediate E. coli isolates. Nine of these isolates did not contain any bla(TEM) genes, whereas the remaining two did. No genes encoding SHV or extended-spectrum beta-lactamases were detected. Two...

  7. Pharmacodynamics of Imipenem in Combination with beta-Lactamase Inhibitor MK7655 in a Murine Thigh Model

    NARCIS (Netherlands)

    Mavridou, E.; Melchers, M.J.B.; Mil, A.C. van; Mangin, E.; Motyl, M.R.; Mouton, J.W.

    2015-01-01

    MK7655 is a newly developed beta-lactamase inhibitor of class A and class C carbapenemases. Pharmacokinetics (PK) of imipenem-cilastatin (IMP/C) and MK7655 were determined for intraperitoneal doses of 4 mg/kg to 128 mg/kg of body weight. MIC and pharmacodynamics (PD) studies of MK7655 were performed

  8. Predicting carriage with extended-spectrum beta-lactamase-producing bacteria at hospital admission : A cross-sectional study

    NARCIS (Netherlands)

    Platteel, T. N.; Leverstein-van Hall, M. A.; Cohen Stuart, J. W.; Thijsen, S. F T; Mascini, E. M.; van Hees, B. C.; Scharringa, J.; Fluit, A. C.; Bonten, M. J M

    2015-01-01

    The prevalence of patients colonized with extended-spectrum beta-lactamase (ESBL)-producing bacteria increases, especially in long-term-care facilities (LTCFs). Identification of ESBL carriers at hospital admission is relevant for infection control measures and antibiotic therapy for nosocomial infe

  9. Survey of metallo-beta-lactamase-producing Enterobacteriaceae colonizing patients in European ICUs and rehabilitation units, 2008-11

    NARCIS (Netherlands)

    Papagiannitsis, C. C.; Izdebski, R.; Baraniak, A.; Fiett, J.; Herda, M.; Hrabak, J.; Derde, L. P. G.; Bonten, M. J. M.; Carmeli, Y.; Goossens, H.; Hryniewicz, W.; Brun-Buisson, C.; Gniadkowski, M.

    2015-01-01

    Objectives: The objective of this study was to perform a multinational survey of patients' colonization by metallo-beta-lactamase (MBL)-producing Enterobacteriaceae, including their molecular characterization. Methods: Patients in 18 hospital units across Europe and Israel (n = 17945) were screened

  10. Emergence of New Delhi Metallo-Beta-Lactamase (NDM-1) and Klebsiella pneumoniae Carbapenemase (KPC-2) in South Africa

    OpenAIRE

    Brink, Adrian J; Coetzee, Jennifer; Clay, Cornelis G.; Sithole, Sindi; Richards, Guy A; Poirel, Laurent; Nordmann, Patrice

    2012-01-01

    This report documents emergence of New Delhi metallo-beta-lactamase (NDM-1) and Klebsiella pneumoniae carbapenemase (KPC-2) in K. pneumoniae and Enterobacter cloacae in South Africa. NDM-1 producers have not been described in South Africa, and this is the first instance that KPC producers have been identified in Africa. The two patients infected with these carbapenemase-producing bacteria demised.

  11. In vitro activities of 15 oral beta-lactams against Klebsiella pneumoniae harboring new extended-spectrum beta-lactamases.

    OpenAIRE

    Kitzis, M D; Liassine, N; Ferré, B.; Gutmann, L; Acar, J F; Goldstein, F.

    1990-01-01

    The activities of 15 oral beta-lactams against Klebsiella pneumoniae harboring new extended-spectrum beta-lactamases were studied. All compounds were affected by these enzymes, especially by the SHV derivatives. Except for ceftibuten, the compounds with the greatest intrinsic activity were more affected by the presence of these enzymes than were older compounds with moderate intrinsic activity.

  12. Evaluation of different phenotypic methods for detection of amp c beta-lactamase producing bacteria in clinical isolates

    International Nuclear Information System (INIS)

    To compare the sensitivity and specificity of different phenotypic methods for detection of Amp C betalactamase producing bacteria. Study Design: Analytical study. Place and Duration of Study: Department of Microbiology, Army Medical College / National University of Sciences and Technology (NUST), Islamabad, Pakistan, from June 2010 to December 2010. Methodology: A total of 150 clinical isolates were screened for presence of Amp C beta-lactamase by using the cefoxitin disc. The confirmatory methods evaluated were inhibitor based assay (boronic acid), Amp C disc test and Amp C Etest. Three dimensional enzyme extract assay was used as the reference method for determining the sensitivity and specificity. Results: Among the total isolates tested, 62.8% bacteria showed the presence of Amp C beta-lactamase by standard three dimensional enzyme extract assay. Among the three methods compared, boronic acid disk test found out to be highly sensitive (88%) and specific (92%) for the detection of Amp C beta-lactamase producing bacteria. Conclusion: Detection of Amp C production is crucial in order to establish the antibiotic therapy and to attain the favourable clinical outcomes. Implementation of simple tests like boronic acid disk tests in the laboratories will help to alleviate the spread of Amp C beta-lactamase harboring organisms. (author)

  13. Prevalence of metallo-beta-lactamase among Pseudomonas aeruginosa and Acinetobacter baumannii in a Korean university hospital and comparison of screening methods for detecting metallo-beta-lactamase.

    Science.gov (United States)

    Oh, Eun-Jee; Lee, Seungok; Park, Yeon-Joon; Park, Jung Jun; Park, Kanggyun; Kim, Sang-Il; Kang, Moon Won; Kim, Byung Kee

    2003-09-01

    To identify the metallo-beta-lactamases (MBLs) prevalent in Korea, a total of 130 clinical isolates of Pseudomonas aeruginosa and Acinetobacter baumannii (99 P. aeruginosa and 31 A. baumannii) with a reduced susceptibility to imipenem (IPM) and/or ceftazidime (CAZ) was subjected to PCR analyses with primers specific to bla(IMP-1), bla(VIM-1), and bla(VIM-2). In addition, inhibitor-potentiated disk diffusion methods (IPD) using two kinds of substrate-inhibitor combinations (ceftazidime-2-mercaptopropionic acid (2MPA) and imipenem-EDTA) were investigated. Thirty-three isolates (29 P. aeruginosa and 4 A. baumannii) carried bla(VIM-2) and two P. aeruginosa isolates harbored bla(IMP-1). The enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) pattern revealed that many of the VIM-2-producing P. aeruginosa isolates were clonally related, whereas the A. baumannii isolates were diverse. The inhibitor-potentiated disk diffusion test using imipenem-EDTA was highly sensitive and specific for detecting the VIM-2 producer. These results suggest that VIM-2 is an important MBL in P. aeruginosa and A. baumannii in the Korean hospital of this study and that the IMP-1-producing P. aeruginosa has also emerged. Screening for MBLs and strict infection control for these isolates will contribute to prevent further spread of resistance. PMID:12842488

  14. Extended-Spectrum-Beta-Lactamases, AmpC Beta-Lactamases and Plasmid Mediated Quinolone Resistance in Klebsiella spp. from Companion Animals in Italy

    DEFF Research Database (Denmark)

    Donati, Valentina; Feltrin, Fabiola; Hendriksen, Rene S.;

    2014-01-01

    We report the genetic characterization of 15 Klebsiella pneumoniae (KP) and 4 isolates of K. oxytoca (KO) from clinical cases in dogs and cats and showing extended-spectrum cephalosporin (ESC) resistance. Extended spectrum beta-lactamase (ESBL) and AmpC genes, plasmid-mediated quinolone resistance...... patterns observed, including two clusters of two (ST340) and four (ST101) indistinguishable isolates, respectively. All isolates harbored at least one ESBL or AmpC gene, all carried on transferable plasmids (IncR, IncFII, IncI1, IncN), and 16/19 were positive for PMQR genes (qnr family or aac(6')-Ib......-cr). The most frequent ESBL was CTX-M-15 (11/19, 58%), detected in all KP ST101, in one KP ST15 and in both KP ST340. bla(CTX-M-15) was carried on IncR plasmids in all but one KP isolate. All KP ST15 isolates harbored different ESC resistance genes and different plasmids, and presented the non-transferable bla...

  15. Activities of beta-lactam antibiotics against Escherichia coli strains producing extended-spectrum beta-lactamases.

    Science.gov (United States)

    Jacoby, G A; Carreras, I

    1990-05-01

    Seven extended-spectrum beta-lactamases related to TEM and four enzymes derived from SHV-1 were transferred to a common Escherichia coli host so that the activity of a variety of beta-lactams could be tested in a uniform genetic environment. For most derivatives, penicillinase activity was 10% or less than that of strains making TEM-1, TEM-2, or SHV-1 beta-lactamase, suggesting that reduced catalytic efficiency accompanied the broader substrate spectrum. Despite this deficit, resistance to aztreonam, carumonam, cefdinir, cefepime, cefixime, cefmenoxime, cefotaxime, cefotiam, cefpirome, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime, and E1040 was enhanced. For strains producing TEM-type enzymes, however, MICs of carumonam, cefepime, cefmenoxime, cefotiam, cefpirome, and ceftibuten were 8 micrograms/ml or less. Susceptibilities of cefmetazole, cefotetan, cefoxitin, flomoxef, imipenem, meropenem, moxalactam, temocillin, FCE 22101, and Sch 34343 were unaffected. FCE 22101, imipenem, meropenem, and Sch 34343 were inhibitory for all strains at 1 microgram/ml or less. In E. coli an OmpF- porin mutation in combination with an extended-spectrum beta-lactamase enhanced resistance to many of these agents, but generally by only fourfold. Hyperproduction of chromosomal AmpC beta-lactamase increased resistance to 7-alpha-methoxy beta-lactams but not that to temocillin. When tested at 8 micrograms/ml, clavulanate was more potent than sulbactam or tazobactam in overcoming resistance to ampicillin, while cefoperazone-sulbactam was more active than ticarcillin-clavulanate or piperacillin-tazobactam, especially against TEM-type extended-spectrum beta-lactamases. PMID:2193623

  16. Caracterização de beta-lactamases em Serratia fonticola

    OpenAIRE

    Henriques, Isabel

    2001-01-01

    A estirpe Serratia fonticola UTAD54 foi isolada no âmbito de um estudo realizado com o objectivo de analisar a presença e disseminação de genes de resistência a antibióticos, em bactérias de águas de consumo. Verificou-se que esta estirpe é resistente a diversos antibióticos do grupo dos beta-lactâmicos, nomeadamente às penicilinas e aos carbapenemos. No âmbito deste estudo foi detectada a produção de uma metalo-beta-lactamase denominada SfhI, responsável pelo fenótipo de re...

  17. [Extended-spectrum beta-lactamases in Klebsiella pneumoniae isolated at the Cordoba Children's Hospital, Argentina].

    Science.gov (United States)

    Saka, H A; Egea, M; Culasso, C; Rollán, R; Avaro, A; Carvajal, L

    2003-01-01

    The aim of the present study was to investigate the presence of extended-spectrum beta-lactamases (ESBL) in Klebsiella pneumoniae isolated at the "Hospital de Niños de Córdoba". The strains were collected from inpatients between January 1996 and July 2000. A total of 150 ESBL producer isolates were detected. During 1996 the prevalence of ESBL producer K. pneumoniae was 20%, but since 1998 the values have increased to approximately 60%. Phenotypic analysis such as isoelectric point (pl) and antibiotyping performed in 32 randomly selected isolates showed two different enzyme profiles: 81% had ESBL with pl = 7.9 and preferential activity against cefotaxime, while 19% showed ESBL with pl = 5.4 and preferential activity against ceftazidime. No isolates resistant to imipenem or ciprofloxacin were detected. Susceptibility to other antimicrobial agents varied, but resistance to gentamicin was strongly associated with ESBL producer isolates. Resistance determinants could be transferred to Escherichia coli by conjugation assays. PMID:12833674

  18. Dynamics and spatial distribution of beta-lactamase expression in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bagge, N.; Hentzer, Morten; Andersen, Jens Bo; Ciofu, O.; Givskov, Michael Christian; Høiby, N.

    2004-01-01

    The development of resistance to beta-lactam antibiotics is a problem in the treatment of chronic Pseudomonas aeruginosa infection in the lungs of patients with cystic fibrosis. The main resistance mechanism is high-level expression of the chromosomally encoded AmpC beta-lactamase of P. aeruginosa...... cells growing in biofilms. Several genes have been shown to influence the level of ampC expression, but little is known about the regulation of ampC expression in P. aeruginosa biofilms. To study the expression of ampC in P. aeruginosa biofilms, we constructed a reporter that consisted of the fusion of...... the ampC promoter to gfp(ASV) encoding an unstable version of the green fluorescent protein. In vitro biofilms of P. aeruginosa were exposed to the beta-lactam antibiotics imipenem and ceftazidime. Sub-MICs of imipenem significantly induced the monitor system of the biofilm bacteria in the peripheries...

  19. Reconstitution of Bacillus cereus 5/B/6 metallo-[beta]-lactamase activity with copper

    Energy Technology Data Exchange (ETDEWEB)

    Hilliard, N.P.; Shaw, R.W. (Texas Tech Univ., Lubbock (United States))

    1992-01-01

    Pathogenic bacteria become resistant to [beta]-lactam antibiotics such as penicillins and cephalosporins through the production of enzymes called [beta]-lactamases. The authors have successfully reconstituted the enzymatic activity of the metallo-[beta]-lactamase of Bacillus cereus 5/B/6 purified from an E. coli expression vector system by the addition of Cu(II) to the apoenzyme. This is the first report that copper supports catalytic activity in this enzyme. Maximal activity of the copper-reconstituted enzyme was achieved by a careful addition of a stoichiometric amount of CuSO[sub 4] to 200 [mu]M apoenzyme. Using either benzylpenicillin or cephalosporin C as the substrate, reconstitution of the activity by addition of copper to the apoenzyme resulted in the recovery of approximately 35% of the control activity of the native Zn(II) enzyme. In agreement with previous reports, in the presence of excess Cu(II), the preparation did not possess measurable catalytic activity. Electronic spectra of the copper-reconstituted enzyme displayed adsorption maxima at 394, 698 and 1,022 nm with extinction coefficients of 2,656, 55 and < 3 M[sup [minus]1]cm[sup [minus]1] respectively. Circular dichorism spectra in the ultraviolent region (UVCD) of the copper-reconstituted enzyme were identical with those of the native Zn(II) enzyme. Addition of excess cephalosporin C to the copper-reconstituted enzyme caused a decrease of about 50% of the absorbance of the 394 nm band and the formation of a new feature at 350 nm.

  20. Extended-spectrum beta-lactamases: implications for the clinical laboratory and therapy.

    Science.gov (United States)

    Harada, Sohei; Ishii, Yoshikazu; Yamaguchi, Keizo

    2008-12-01

    Production of extended-spectrum beta-lactamase (ESBL) is one of the most important resistance mechanisms that hamper the antimicrobial treatment of infections caused by Enterobacteriaceae. ESBLs are classified into several groups according to their amino-acid sequence homology. While TEM and SHV enzymes were the most common ESBLs in the 1990s, CTX-M enzymes have spread rapidly among Enterobacteriaceae in the past decade. In addition, some epidemiological studies showed that organisms producing CTX-M enzymes had become increasingly prevalent in the community setting in certain areas in the world. Several novel enzymes with hydrolyzing activity against oxyimino-cephalosporins, albeit with additional enzymatic characteristics different from those of original TEM and SHV ESBLs (e.g., inhibitor-resistance), have been discovered and pose a problem on the definition of ESBLs. Although several methods to detect the production of ESBL are available in clinical laboratories, existence of other factors contributing resistance against beta-lactams, e.g., inducible production of Amp-C beta-lactamase by some species of Enterobacteriaceae, or inhibitor-resistance in some ESBLs may hinder the detection of ESBLs with these methods. Carbapenems are stable against hydrolyzing activity of ESBLs and are regarded as the drug of choice for the treatment of infections caused by ESBL-producing Enterobacteriaceae. Although several other antimicrobial agents, such as fluoroquinolones and cephamycins, may have some role in the treatment of mild infections due to those organisms, clinical data that warrant the use of antimicrobial agents other than carbapenems in the treatment of serious infections due to those organisms are scarce for now. PMID:19127103

  1. [Detection of bla(CTX-M) beta-lactamase genes in extended-spectrum beta-lactamase producing gram-negative bacteria].

    Science.gov (United States)

    Bayraktar, Banu; Toksoy, Buket; Bulut, Emin

    2010-04-01

    Widespread production of CTX-M type extended-spectrum beta-lactamases (ESBL) in Enterobacteriaceae strains which are resistant to extended-spectrum cephalosporins is the most remarkable example for rapid and global spread of plasmid mediated antimicrobial resistance in bacteria. Consecutive 200 ESBL producing Enterobacteriaceae strains out of 1640 isolates that were obtained from clinical samples (167 urine, 11 wound, 7 bronchoalveolar lavage, 3 peritoneal fluid, 2 cerebrospinal fluid, 2 biopsy, 2 tracheal aspirate, 2 conjunctiva, 1 abscess, 1 catheter) between February to July 2009 in our laboratory were included to this study. Among the 200 ESBL positive isolates 141 (70.5%) were Escherichia coli, 51 (26%) were Klebsiella pneumoniae, 5 (2.5%) were Enterobacter spp. and one of each (0.5%) Citrobacter freundii, Klebsiella oxytoca and Proteus mirabilis. ESBL positivity was 11% among the 123 community-acquired strains and 13% among the 77 hospital acquired strains, the statistical difference being insignificant (p > 0.05). The prevalence of bla(CTX-M) beta-lactamase genes were detected by multiplex polymerase chain reaction with the use of two general primer sets: CTX-MA1 and CTX-MA2 primers for the amplification of CTX-M-1, CTX-M-2 and CTX-M-9 enzymes group, and CTX825-F and CTX825-R primers for the amplification of CTX-M-8 and CTX-M-25 enzymes group. bla(CTX-M) genes were detected in 167 out of 200 strains (83.5%). CTX-M production rates in community and hospital acquired strains were found as 86.2% and 79.2%, respectively and no statistically significant difference was detected (p > 0.05). CTX-M producing strains were either E. coli (n = 132) or Klebsiella spp. (n = 35) and were expressing one of the enzymes from CTX-M-1, CTX-M-2 or CTX-M-9 groups. No strains carrying CTX-M-8 or CTX-M-25 group enzymes were detected. CTX-M production rates in ESBL producing E. coli strains in community and hospital were found as 92.5% and 95.7%, respectively, whereas the same rates

  2. Prevalence of Multidrug Resistant Extended-Spectrum Beta-Lactamase Producing Gram-Negative Bacteria in Neonatal Sepsis

    Directory of Open Access Journals (Sweden)

    Ali Peirovifar

    2014-02-01

    Full Text Available Objectives: Neonatal sepsis with extended-spectrum beta-lactamase (ESBL producing microorganisms is recognized increasingly in recent years. ESBL can be produced by various bacterial strains. This study was conducted to determine the prevalence of ESBL producing pathogens in neonatal sepsis and its impact on clinical outcome. Materials and Methods: A study was carried out from Jan 2012 to Jan 2013 in a referral university hospital. All neonates who had diagnosed as sepsis were enrolled in this study. Blood cultures were processed using the automated BACTEC 9120 system. Antibiotic resistance and beta-lactamase production of bacterial isolates was tested. All patients were followed till discharge. Results: One hundred three neonates with gestation age 36.7±3.2 weeks were enrolled in this study and 56 cases (54% were boys. The most common isolated gram negative pathogens were Klebsiella pneumoniae, Acinetobacter species, and Pseudomonas aeruginosa. The rate for beta-lactamase production were 97.7% in Klebsiella pneumoniae , 81.3% in Acinetobacter, 85.7% in E. coli, 53.3% in Pseudomonas aeruginosa and 100% in Serratia. Thirty eight (35.9% neonates were dead, that 34 of them were beta-lactamase producers. The mean duration of hospitalization were longer in patients infected with beta-lactamase producers (30.2±20.5 vs. 22.8±16.6 days P=0.05 and ESBL producing strains (29.13±20.39 vs. 19±9.84 P=0.05. ESBL production rate were determined 95.5% and 86.7% in Klebsiella pneumoniae by combined disk test (CDT and double disk synergy test (DDST method, respectively. These methods were positive for ESBL production in 78.6% and 64.3% of E. coli isolates, respectively. Conclusion: in our study, the high rate of beta-lactamase and ESBL production were determined for common isolated organisms in neonatal sepsis. Infection with ESBL producing pathogens was associated with longer hospital stay. CDT method was detected more ESBL producing pathogens than DDST

  3. Chemoprophylactic efficacy against experimental endocarditis caused by beta-lactamase-producing, aminoglycoside-resistant enterococci is associated with prolonged serum inhibitory activity.

    OpenAIRE

    Bayer, A S; Tu, J

    1990-01-01

    We studied the prevention of experimental aortic endocarditis caused by a beta-lactamase-producing, aminoglycoside-resistant strain of Enterococcus faecalis (HH22) in 146 catheterized rabbits. Both vancomycin and ampicillin-sulbactam readily killed this resistant enterococcus strain in vitro. At a challenge inoculum of approximately 10(9) CFU, vancomycin (40 mg/kg intravenously [i.v.]), ampicillin (40 mg/kg i.v.), or a combination of ampicillin plus a beta-lactamase inhibitor, sulbactam (20 m...

  4. A step towards the discrimination of beta-lactamase-producing clinical isolates of Enterobacteriaceae and Pseudomonas aeruginosa by MALDI-TOF mass spectrometry

    OpenAIRE

    Schaumann, Reiner; Knoop, Nicolas; Genzel, Gelimer H; Losensky, Kevin; Rosenkranz, Christiane; Stîngu, Catalina S; Schellenberger, Wolfgang; Rodloff, Arne C; Eschrich, Klaus

    2012-01-01

    Summary Background Matrix-Assisted Laser-Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) has already proven to be a powerful tool for species identification in microbiological laboratories. As adequate and rapid screening methods for antibiotic resistance are crucially needed, the present study investigated the discrimination potential of MALDI-TOF MS among extended-spectrum-beta-lactamase (ESBL) or metallo-beta-lactamases- (MBL) producing and the nonproducing strains of...

  5. 6 beta-Iodopenicillanic acid (UI-38,006), a beta-lactamase inhibitor that extends the antibacterial spectrum of beta-lactam compounds: initial bacteriological characterization.

    OpenAIRE

    Moore, B A; Brammer, K W

    1981-01-01

    UK-38,006, 6 beta-iodopenicillanic acid, was shown to be a potent inhibitor of beta-lactamase enzymes. It potentiated the antibacterial action of ampicillin in vitro against beta-lactamase-producing strains of Staphylococcus aureus, Haemophilus influenzae, Bacteroides fragilis. Neisseria gonorrhoeae, and many Enterobacteriaceae. This ability to synergize with ampicillin was also demonstrated in vivo after oral administration of UK-38,006 to experimentally infected mice. UK-38,006 was also sho...

  6. Use of the chromosomal class A beta-lactamase of Mycobacterium fortuitum D316 to study potentially poor substrates and inhibitory beta-lactam compounds.

    OpenAIRE

    Galleni, M; Franceschini, N; Quinting, B; Fattorini, L.; Orefici, G.; Oratore, A; Frère, J M; Amicosante, G

    1994-01-01

    Sixteen different compounds usually considered beta-lactamase stable or representing potential beta-lactam inhibitors and inactivators were tested against the beta-lactamase produced by Mycobacterium fortuitum. The compounds exhibiting the most interesting properties were BRL42715, which was by far the best inactivator, and CGP31608 and ceftazidime, which were not recognized by the enzyme. These compounds thus exhibited adequate properties for fighting mycobacterial infections. Although cloxa...

  7. Hospital outbreak of carbapenem-resistant Pseudomonas aeruginosa producing VIM-1, a novel transferable metallo-beta-lactamase.

    Science.gov (United States)

    Cornaglia, G; Mazzariol, A; Lauretti, L; Rossolini, G M; Fontana, R

    2000-11-01

    A total of 8 Pseudomonas aeruginosa isolates was collected from 7 different patients in different wards of the University Hospital of Verona, Italy, from February 1997 to February 1998. The high level of resistance to carbapenems (imipenem minimum inhibitory concentration was always >128 microg/mL) and other broad-spectrum beta-lactams and the rate of imipenem hydrolysis and its inhibition by ethylenediamine-tetra-acetic acid were all suggestive of production of a carbapenem-hydrolyzing metallo-beta-lactamase. A specific DNA probe derived from the recently cloned bla(VIM-1) gene hybridized to all the isolates. A genomic DNA fingerprinting profile revealed clonal relatedness for 7 of 8 isolates. A description of this hospital outbreak is reported, the occurrence of which confirms that proliferation of metallo-beta-lactamase-producing strains multiply resistant to beta-lactams is already a reality outside Japan. These findings emphasize the need for early recognition of similar isolates. PMID:11073738

  8. High beta-Lactamase Levels Change the Pharmacodynamics of beta-Lactam Antibiotics in Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Wang, Hengzhuang; Ciofu, Oana; Yang, Liang;

    2013-01-01

    , microtiter plates, and on alginate beads were treated with different concentrations of ceftazidime and imipenem. The kinetics of antibiotics on the biofilms was investigated in vitro by time-kill methods. Time-dependent killing of ceftazidime was observed in PAO1 biofilms, but concentration-dependent killing......Resistance to beta-lactam antibiotics is a frequent problem in Pseudomonas aeruginosa lung infection of cystic fibrosis (CF) patients. This resistance is mainly due to the hyperproduction of chromosomally encoded beta-lactamase and biofilm formation. The purpose of this study was to investigate the...... activity of ceftazidime was observed for beta-lactamase-overproducing biofilms of P. aeruginosa in all three models. Ceftazidime showed time-dependent killing on planktonic PAO1 and PA Delta DDh2Dh3. This difference is probably due to the special distribution and accumulation in the biofilm matrix of beta...

  9. Commensal Enterobacteriaceae as reservoirs of extended-spectrum beta-lactamases, integrons, and sul genes in Portugal

    OpenAIRE

    Machado, Elisabete; Coque, Teresa M.; Cantón, Rafael; Sousa, João C.; Peixe, Luísa

    2013-01-01

    Bacteria colonizing the human intestine have a relevant role in the spread of antimicrobial resistance. We investigated the faecal carriage of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in healthy humans from Portugal and analyzed the distribution of sul genes and class 1 and 2 integrons. Faecal samples (n = 113) were recovered from healthy persons (North/Centre of Portugal, 2001–2004) and plated on MacConkey agar with and without ceftazidime (1 mg/L) or cefotaxime (...

  10. Prevalence of Multidrug Resistant Extended-Spectrum Beta-Lactamase Producing Gram-Negative Bacteria in Neonatal Sepsis

    OpenAIRE

    Ali Peirovifar; Mohammad Ahangarzadeh Rezaee; Manizheh Mostafa Gharehbaghi

    2014-01-01

    Objectives: Neonatal sepsis with extended-spectrum beta-lactamase (ESBL) producing microorganisms is recognized increasingly in recent years. ESBL can be produced by various bacterial strains. This study was conducted to determine the prevalence of ESBL producing pathogens in neonatal sepsis and its impact on clinical outcome. Materials and Methods: A study was carried out from Jan 2012 to Jan 2013 in a referral university hospital. All neonates who had diagnosed as sepsis were enrolled in...

  11. SCO-1, a novel plasmid-mediated class A beta-lactamase with carbenicillinase characteristics from Escherichia coli.

    Science.gov (United States)

    Papagiannitsis, C C; Loli, A; Tzouvelekis, L S; Tzelepi, E; Arlet, G; Miriagou, V

    2007-06-01

    A novel class A beta-lactamase (SCO-1) encoded by an 80-kb self-transferable plasmid from Escherichia coli is described. The interaction of SCO-1 with beta-lactams was similar to that of the CARB-type enzymes. Also, SCO-1 exhibited a 51% amino acid sequence identity with the RTG subgroup of chromosomal carbenicillinases (RTG-1, CARB-5, and CARB-8). PMID:17353248

  12. Differences in Extended-Spectrum Beta-Lactamase Producing Escherichia coli Virulence Factor Genes in the Baltic Sea Region

    OpenAIRE

    Jana Lillo; Kristiine Pai; Arta Balode; Mariia Makarova; Kristi Huik; Siiri Kõljalg; Marina Ivanova; Lidia Kaftyreva; Jolanta Miciuleviciene; Paul Naaber; Kristel Parv; Anastasia Pavelkovich; Tiiu Rööp; Karolin Toompere; Ludmila Suzhaeva

    2014-01-01

    The aim of this study was to compare the prevalence of different virulence factor (VF) genes in extended-spectrum beta-lactamase (ESBL) producing Escherichia coli strains isolated from the Baltic Sea region. A total of 432 strains of phenotypically ESBL positive E. coli were collected from 20 institutions located in Estonia, Latvia, Lithuania, and the region of St. Petersburg in Russia from January to May 2012 and analyzed for phylogenetic group and prevalence of 23 VF genes. The strains were...

  13. Efficacy of oxacillin and ampicillin-sulbactam combination in experimental endocarditis caused by beta-lactamase-hyperproducing Staphylococcus aureus.

    OpenAIRE

    Thauvin-Eliopoulos, C; Rice, L B; Eliopoulos, G M; Moellering, R C

    1990-01-01

    Optimal therapy of infections caused by borderline oxacillin-susceptible, beta-lactamase-hyperproducing Staphylococcus aureus has not been established. We used a rat model of aortic valve endocarditis to examine efficacies of antibiotic regimens against a borderline oxacillin-susceptible strain as compared with a fully susceptible S. aureus strain. Animals were treated with oxacillin alone or in combination with sulbactam or with ampicillin-sulbactam combinations at two dose levels. Infection...

  14. Fecal carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae in swine and cattle at slaughter in Switzerland

    OpenAIRE

    Geser, N.; Stephan, R; Kuhnert, P.; R. Zbinden; Kaeppeli, U.; Cernela, N; Haechler, H.

    2011-01-01

    During the past decade, extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have become a matter of great concern in human medicine. ESBL-producing strains are found in the community, not just in hospital-associated patients, which raises a question about possible reservoirs. Recent studies describe the occurrence of ESBL-producing Enterobacteriaceae in meat, fish, and raw milk; therefore, the impact of food animals as reservoirs for and disseminators of such strains into the...

  15. Molecular epidemiology of hybrid CTX-M type beta-lactamases among Escherichia coli isolates from human and animals

    OpenAIRE

    Liu, Chunjiao; 刘春娇

    2014-01-01

    A novel and effective mechanism of forming resistance to beta-lactam antibiotics by producing hybrid CTX-M-type beta-lactamases through genetic recombination was reported recently, albeit sporadically. Its primary introductory factor is the horizontal resistant gene transfer that occurs within the 876 bp 〖bla〗_(CTX-M) gene in Escherichia coli isolates, followed by the extensive application of beta-lactam antimicrobials. 〖bla〗_(CTX-M) originated from the nonpathogenic commensal Kluyvera spp., ...

  16. Multidrug resistance found in extended-spectrum beta-lactamase-producing Enterobacteriaceae from rural water reservoirs in Guantao, China

    OpenAIRE

    Zhang, Hongna; Zhou, Yufa; Guo, Shuyuan; Chang, Weishan

    2015-01-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have been isolated from humans and animals across the world. However, data on prevalence of ESBL-producing Enterobacteriaceae from rural water reservoirs is limited. This study aimed to isolate and characterize ESBL-producing Enterobacteriaceae in rural water reservoirs in Guantao, China. ESBL-producing Enterobacteriaceae were found in 5 (16.7%) of 30 sampled rural water reservoirs. Sixty-six individual isolates expressing a...

  17. In vitro activity of LK-157, a novel tricyclic carbapenem as broad-spectrum {beta}-lactamase inhibitor.

    Science.gov (United States)

    Paukner, Susanne; Hesse, Lars; Prezelj, Andrej; Solmajer, Tomaz; Urleb, Uros

    2009-02-01

    LK-157 is a novel tricyclic carbapenem with potent activity against class A and class C beta-lactamases. When tested against the purified TEM-1 and SHV-1 enzymes, LK-157 exhibited 50% inhibitory concentrations (IC(50)s) in the ranges of the clavulanic acid and tazobactam IC(50)s (55 nM and 151 nM, respectively). Moreover, LK-157 significantly inhibited AmpC beta-lactamase (IC(50), 62 nM), as LK-157 was >2,000-fold more potent than clavulanic acid and approximately 28-fold more active than tazobactam. The in vitro activities of LK-157 in combination with amoxicillin, piperacillin, ceftazidime, cefotaxime, ceftriaxone, cefepime, cefpirome, and aztreonam against an array of Ambler class A (TEM-, SHV-, CTX-M-, KPC-, PER-, BRO-, and PC-type)- and class C-producing bacterial strains derived from clinical settings were evaluated in synergism experiments and compared with those of clavulanic acid, tazobactam, and sulbactam. In vitro MICs against ESBL-producing strains (except CTX-M-containing strains) were reduced 2- to >256-fold, and those against AmpC-producing strains were reduced even up to >32-fold. The lowest MICs (combination of cefepime and cefpirome with a constant LK-157 concentration of 4 microg/ml, thus raising an interest for further development. LK-157 proved to be a potent beta-lactamase inhibitor, combining activity against class A and class C beta-lactamases, which is an absolute necessity for use in the clinical setting due to the worldwide increasing prevalence of bacterial strains resistant to beta-lactam antibiotics. PMID:19075067

  18. Detection Of Extended-Spectrum Beta Lactamase in Klebsiella pneumoniae and Klebsiella oxytoca Bacteria with the Combined Disc Method

    OpenAIRE

    Ebru Yılmaz; Güven Uraz

    2011-01-01

    Extended-spectrum beta lactamases (ESBLs) are responsible for resistance to cephalosporins (ceftazidime, ceftriaxone, and cefotaxime) and aztreonam in gram-negative bacilli. ESBL producing Klebsiella bacteria are a major problem for clinicians, ESBLs increase are cause of failure in treatment particularly paediatric patients and also in medical and surgical units. In this research ESBL was investigated by combined disc method. In this research, 128 clinical isolates of Klebsiella ssp. were co...

  19. High prevalence rate of extended-spectrum beta-lactamases (ESBL) among Enterobacteriaceae in a small Brazilian public hospital

    OpenAIRE

    Adriane Lenhard-Vidal; Rosilene Fressatti Cardoso; Rubia Andreia Falleiros de Pádua; Vera Lúcia Dias Siqueira

    2011-01-01

    The production of extended-spectrum beta-lactamases (ESBL) is considered one of the most important resistance mechanisms that impair antimicrobial treatment of infections caused by Enterobacteriaceae. Data on culture and susceptibility tests were collected from the Clinical Analyses and Research Laboratory charts reporting on patients admitted to the University Hospital of Maringá (HUM) from January 2004 to December 2009. The following Enterobacteriaceae were selected: Escherichia coli, Klebs...

  20. Detection Of Extended-Spectrum Beta Lactamase in Klebsiella pneumoniae and Klebsiella oxytoca Bacteria with the Combined Disc Method

    OpenAIRE

    Yılmaz, Ebru; Uraz, Güven

    2000-01-01

    Extended-spectrum beta lactamases (ESBLs) are responsible for resistance to cephalosporins (ceftazidime, ceftriaxone, and cefotaxime) and aztreonam in gram-negative bacilli. ESBL producing Klebsiella bacteria are a major problem for clinicians, ESBLs increase are cause of failure in treatment particularly paediatric patients and also in medical and surgical units. In this research ESBL was investigated by combined disc method. In this research, 128 clinical isolates of Klebsiella ssp. were co...

  1. A novel extended-spectrum TEM-type beta-lactamase, TEM-138, from Salmonella enterica serovar Infantis.

    OpenAIRE

    chouchani, chedli; Berlemont, Renaud; A. Masmoudi; Galleni, Moreno; Frère, Jean-Marie; O Belhadj; Ben-Mahrez, K.

    2006-01-01

    A novel natural TEM beta-lactamase with extended-spectrum activity, TEM-138, was identified in a ceftazidime-resistant clinical isolate of Salmonella enterica serovar Infantis. Compared to TEM-1, TEM-138 contains the following mutations: E104K, N175I, and G238S. The bla(TEM-138) gene was located on a 50-kb transferable plasmid. Expression studies with Escherichia coli revealed efficient ceftazidimase and cefotaximase activities for TEM-138.

  2. Laboratory Detection of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae: Evaluation of Two Screening Agar Plates and Two Confirmation Techniques▿

    OpenAIRE

    Overdevest, I. T. M. A.; Willemsen, I; Elberts, S.; Verhulst, C; Kluijtmans, J.A.J.W.

    2011-01-01

    The worldwide prevalence of extended-spectrum-beta-lactamase-producing ESBL-producing Enterobacteriaceae (ESBL-E) is increasing, making the need for optimized detection techniques more urgent. In this study we investigated the performance of two ESBL-E screening and two ESBL-E confirmation techniques. In accordance with the Dutch national guidelines (www.wip.nl), a collection of 642 highly resistant Enterobacteriaceae strains, as identified by Vitek2, was used to test the performances of two ...

  3. The KPC type beta-lactamases: new enzymes that confer resistance to carbapenems in Gram-negative bacilli.

    OpenAIRE

    Piotr Wieczorek; Jadwiga Jaworowska; Alina Ostas; Paweł Sacha; Dominika Ojdana; Jerzy Ratajczak; Elzbieta Tryniszewska

    2010-01-01

    Antimicrobial resistance due to the continuous selective pressure from widespread use of antimicrobials in humans, animals and agriculture has been a growing problem for last decades. KPC beta-lactamases hydrolyzed beta-lactams of all classes. Especially, carbapenem antibiotics are hydrolyzed more efficiency than other beta-lactam antibiotics. The KPC enzymes are found most often in Enterobacteriaceae. Recently, these enzymes have been found in isolates of Pseudomonas aeruginosa and Acinetoba...

  4. Role of beta-lactamase in in vivo development of ceftazidime resistance in experimental Pseudomonas aeruginosa endocarditis.

    OpenAIRE

    Bayer, A S; Peters, J.; Parr, T R; Chan, L; Hancock, R E

    1987-01-01

    Two ceftazidime-resistant variants of Pseudomonas aeruginosa (PA-48, PA-60), obtained from cardiac vegetations of rabbits with endocarditis receiving ceftazidime therapy, were studied for mechanisms of resistance. Both resistant variants were stably derepressed for the type Id beta-lactamase, which was ceftazidime inducible in the parental strain (PA-96) used to initially infect the rabbits. There was no evidence of ceftazidime bioinactivation by the resistant strains, and their outer membran...

  5. Microbial resistance and frequency of extended-spectrum beta-lactamase (ESBL) in isolated from blood cultures

    OpenAIRE

    Ruan Carlos Gomes da Silva; Amanda Cristina de Oliveira Silva; Sibele Ribeiro de Oliveira

    2014-01-01

    Introduction:The emergence and spread of isolated carriers of extended-spectrum beta-lactamase (ESBL) have complicated the treatment of nosocomial infections, since its production is not easily identified by the sensitivity tests, routinely performed in clinical laboratories, leading to difficulties in the hospital control of resistant microorganisms and antibiotics misuse.Objective:The objective of this study was to analyze the resistance profile and the frequency of ESBL in Gram-negative ba...

  6. Extended-spectrum-beta-lactamases, AmpC beta-lactamases and plasmid mediated quinolone resistance in klebsiella spp. from companion animals in Italy.

    Directory of Open Access Journals (Sweden)

    Valentina Donati

    Full Text Available We report the genetic characterization of 15 Klebsiella pneumoniae (KP and 4 isolates of K. oxytoca (KO from clinical cases in dogs and cats and showing extended-spectrum cephalosporin (ESC resistance. Extended spectrum beta-lactamase (ESBL and AmpC genes, plasmid-mediated quinolone resistance (PMQR and co-resistances were investigated. Among KP isolates, ST101 clone was predominant (8/15, 53%, followed by ST15 (4/15, 27%. ST11 and ST340, belonging to Clonal Complex (CC11, were detected in 2012 (3/15, 20%. MLST on KP isolates corresponded well with PFGE results, with 11 different PFGE patterns observed, including two clusters of two (ST340 and four (ST101 indistinguishable isolates, respectively. All isolates harbored at least one ESBL or AmpC gene, all carried on transferable plasmids (IncR, IncFII, IncI1, IncN, and 16/19 were positive for PMQR genes (qnr family or aac(6'-Ib-cr. The most frequent ESBL was CTX-M-15 (11/19, 58%, detected in all KP ST101, in one KP ST15 and in both KP ST340. blaCTX-M-15 was carried on IncR plasmids in all but one KP isolate. All KP ST15 isolates harbored different ESC resistance genes and different plasmids, and presented the non-transferable blaSHV-28 gene, in association with blaCTX-M-15, blaCTX-M-1 (on IncR, or on IncN, blaSHV-2a (on IncR or blaCMY-2 genes (on IncI1. KO isolates were positive for blaCTX-M-9 gene (on IncHI2, or for the blaSHV-12 and blaDHA-1 genes (on IncL/M. They were all positive for qnr genes, and one also for the aac(6'-Ib-cr gene. All Klebsiella isolates showed multiresistance towards aminoglycosides, sulfonamides, tetracyclines, trimethoprim and amphenicols, mediated by strA/B, aadA2, aadB, ant (2"-Ia, aac(6'-Ib, sul, tet, dfr and cat genes in various combinations. The emergence in pets of multidrug-resistant Klebsiella with ESBL, AmpC and PMQR determinants, poses further and serious challenges in companion animal therapy and raise concerns for possible bi-directional transmission between

  7. In vitro Efficacy of Meropenem, Colistin and Tigecycline Against the Extended Spectrum Beta-Lactamase Producing Gram Negative Bacilli

    International Nuclear Information System (INIS)

    Objective:To compare the in vitroefficacy of meropenem, colistin and tigecycline against extended spectrum Betalactamase producing Gram negative bacilli by minimal inhibitory concentration. Study Design:Cross-sectional descriptive study. Place and Duration of Study: Department of Microbiology, Army Medical College, National University of Sciences and Technology, Rawalpindi, from June to December 2010. Methodology: Routine clinical specimens were subjected to standard microbiological procedures and the isolates were identified to species level. Extended spectrum beta-lactamase producing Gram negative bacilli were detected by Jarlier disc synergy method and confirmed by ceftazidime and ceftazidime-clavulanate Etest. Minimum Inhibitory Concentration (MIC90) of meropenem, colistin and tigecycline was determined by Etest (AB BIOMERIUX) and the results were interpreted according to the manufacturer's instructions and Clinical and Laboratory Standards Institute guidelines and Food and Drug Authority recommendations. Results were analyzed by using Statistical Package for the Social Sciences version 20. Results: A total of 52 non-duplicate extended spectrum Beta-lactamase-producing Gram negative bacilli were included in the study. The MIC90 of tigecycline (0.75 micro g/ml) was lowest as compared to the meropenem (2 micro g/ml) and colistin (3 micro g/ml). Conclusion: Tigecycline is superior in efficacy against the extended spectrum Beta-lactamase producing Gram negative bacilli as compared to colistin and meropenem. (author)

  8. Nosocomial infections caused by multidrug-resistant isolates of pseudomonas putida producing VIM-1 metallo-beta-lactamase.

    Science.gov (United States)

    Lombardi, Gianluigi; Luzzaro, Francesco; Docquier, Jean-Denis; Riccio, Maria Letizia; Perilli, Mariagrazia; Colì, Alessandra; Amicosante, Gianfranco; Rossolini, Gian Maria; Toniolo, Antonio

    2002-11-01

    Successful carbapenem-based chemotherapy for the treatment of Pseudomonas infections has been seriously hindered by the recent appearance of IMP- and VIM-type metallo-beta-lactamases, which confer high-level resistance to carbapenems and most other beta-lactams. Recently, multidrug-resistant Pseudomonas putida isolates for which carbapenem MICs were >/=32 micro g/ml were recovered from cultures of urine from three inpatients in the general intensive care unit of the Ospedale di Circolo, Varese, Italy. Enzyme assays revealed production of a metallo-beta-lactamase activity, while molecular analysis detected in each isolate a bla(VIM-1) determinant carried by an apparently identical medium-sized plasmid. Conjugation experiments were unsuccessful in transferring the beta-lactamase determinant to Escherichia coli or Pseudomonas aeruginosa. Macrorestriction analysis by pulsed-field gel electrophoresis demonstrated that the isolates were of clonal origin. PCR mapping and sequencing of the variable region of the plasmid-borne class 1 integron carrying the bla(VIM-1) determinant (named In110) showed that the bla(VIM-1)-containing cassette was identical to that previously found in strains of different species from other Italian hospitals and that the cassette array of In110 was not identical but clearly related to that of In70 (a bla(VIM-1)-containing plasmid-borne integron from an Achromobacter xylosoxidans isolate), pointing to a common origin of this cassette and to a related evolutionary history of their cognate integrons. PMID:12409373

  9. Prevalence of extended-spectrum beta-lactamases among Enterobacteriaceae isolated from blood culture in a tertiary care hospital

    International Nuclear Information System (INIS)

    To determine the prevalence of extended spectrum beta-lactamase among Enterobacteriaceae isolated from blood culture in a tertiary care hospital. We carried out this study at the Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia during the period between January 2003 - December 2004. We tested a total of 601 isolates of the family Enterobacteriaceae from blood culture for the prevalence of extended spectrum beta-lactamase (ESBL) production by the standardized disc diffusion method and confirmed by the ESBL E test strips. Ninety-five (15.8%) of the isolates were ESBL producers. Among these, 48.4% were Klebsiella pneumoniae (K. pneumoniae) followed by15.8% of both Escherichia coli (E. coli) and Enterobacter cloacae (Ent. cloacae). Other isolates produced ESBL in low numbers. Klebsiella pneumoniae produced ESBL in significant numbers. Extended spectrum beta-lactamase gram-negative bacilli present significant diagnostic and therapeutic challenges to the management of infections due to these organisms. Microbiology laboratories should start reporting ESBL producing Enterobacteriaceae organism due to their importance in respect to antibiotic therapy and infection control aspects. (author)

  10. Noncovalent Interaction Energies in Covalent Complexes: TEM-1 beta-Lactamase and beta-Lactams

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Xiaojun; Minasov, George; Shoichet, Brian K. (NWU)

    2010-03-08

    The class A {beta}-lactamase TEM-1 is a key bacterial resistance enzyme against {beta}-lactam antibiotics, but little is known about the energetic bases for complementarity between TEM-1 and its inhibitors. Most inhibitors form a covalent adduct with the catalytic Ser70, making the measurement of equilibriumconstants, and hence interaction energies, technically difficult. This study evaluates noncovalent interactions withincovalent complexes by examining the differential stability of TEM-1 and its inhibitor adducts. The thermal denaturation of TEM-1 follows a two-state, reversible model with a melting temperature (T{sub m}) of 51.6 C and a van't Hoff enthalpy of unfolding ({Delta}H{sub VH}) of 146.2 kcal/mol at pH 7.0. The stability of the enzyme changes on forming an inhibitor adduct. As expected, some inhibitors stabilize TEM-1; transition-state analogues increase the T{sub m} by up to 3.7 C(1.7 kcal/mol). Surprisingly, all {beta}-lactam covalent acyl-enzyme complexes tested destabilize TEM-1 significantly relative to the apoenzyme. For instance, the clinically used inhibitor clavulanic acid and the {beta}-lactamase-resistant {beta}-lactams moxalactam and imipenem destabilize TEM-1 by over 2.6 C (1.2 kcal/mol) in their covalent adducts. Based on the structure of the TEM-1/imipenem complex (Maveyraud et al., J Am Chem Soc 1998;120:9748-52), destabilization by moxalactam and imipenem is thought to be caused by a steric clash between the side-chain of Asn132 and the 6(7)-{alpha} group of these {beta}-lactams. To test this hypothesis, the mutant enzyme N132A was made. In contrast with wild-type, the covalent complexes between N132A and both imipenem and moxalactam stabilize the enzyme, consistent with the hypothesis. To investigate the structural bases of this dramatic change instability, the structure of N132A/imipenem was determined by X-ray crystallography. In the complex with N132A, imipenemadopts a very different conformation from that observed in the wild

  11. Evolutionary trajectories of beta-lactamase CTX-M-1 cluster enzymes: predicting antibiotic resistance.

    Directory of Open Access Journals (Sweden)

    Angela Novais

    2010-01-01

    Full Text Available Extended-spectrum beta-lactamases (ESBL constitute a key antibiotic-resistance mechanism affecting Gram-negative bacteria, and also an excellent model for studying evolution in real time. A shift in the epidemiology of ESBLs is being observed, which is characterized by the explosive diversification and increase in frequency of the CTX-M-type beta-lactamases in different settings. This provides a unique opportunity for studying a protein evolutionary radiation by the sequential acquisition of specific mutations enhancing protein efficiency and fitness concomitantly. The existence of driver antibiotic molecules favoring protein divergence has been investigated by combining evolutionary analyses and experimental site-specific mutagenesis. Phylogenetic reconstruction with all the CTX-M variants described so far provided a hypothetical evolutionary scenario showing at least three diversification events. CTX-M-3 was likely the enzyme at the origin of the diversification in the CTX-M-1 cluster, which was coincident with positive selection acting on several amino acid positions. Sixty-three CTX-M-3 derivatives containing all combinations of mutations under positively selected positions were constructed, and their phenotypic efficiency was evaluated. The CTX-M-3 diversification process can only be explained in a complex selective landscape with at least two antibiotics (cefotaxime and ceftazidime, indicating the need to invoke mixtures of selective drivers in order to understand the final evolutionary outcome. Under this hypothesis, we found congruent results between the in silico and in vitro analyses of evolutionary trajectories. Three pathways driving the diversification of CTX-M-3 towards the most complex and efficient variants were identified. Whereas the P167S pathway has limited possibilities of further diversification, the D240G route shows a robust diversification network. In the third route, drift may have played a role in the early stages of

  12. In vitro activity of fosfomycin tromethamine against extended spectrum beta-lactamase producing urinary tract bacteria

    International Nuclear Information System (INIS)

    To determine the in vitro activity of Fosfomycin tromethamine against extended spectrum beta-lactamase producing uropathogens. Study Design: Experimental study. Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from October 2011 to October 2012. Methodology: A total of 381 culture positive ESBL producing isolates from 2400 urine samples submitted over a period of one year were included in this study. Identification of isolates was done by standard biochemical profile of the organisms. The antimicrobial susceptibility of culture positive isolates was performed by disk diffusion method as recommended by Clinical Laboratory Standard Institute guidelines (CLSI). Results: The antimicrobial activity of Fosfomycin to various isolates revealed that 93% of E. coli, 64% Klebsiella spp. 50% Proteus spp. 75% Enterobacter cloacae, 100% Citrobacter freundii, 100% Burkholderia spp. 100% Serratia spp. and 50% Stenotrophomonas maltophilia were susceptible to this chemical compound. Conclusion: Fosfomycin showed excellent effectiveness to most of the common ESBL producing bacteria such as E. coli, Klebsiella and Proteus spp. (author)

  13. Extended-spectrum beta-lactamase-producing Shigella strains in Israel, 2000-2004.

    Science.gov (United States)

    Vasilev, V; Japheth, R; Yishai, R; Andorn, N; Valinsky, L; Navon-Venezia, S; Chmelnitsky, I; Carmeli, Y; Cohen, D

    2007-03-01

    Routine susceptibility testing of 5,616 Shigella isolates at the National Shigella Reference Centre in Israel over a 5-year period (2000-2004) revealed resistance to ceftriaxone in one strain of Shigella boydii 2 and in two strains each of Shigella flexneri 2a, S. flexneri 6, and Shigella sonnei. All seven isolates were confirmed as producers of extended-spectrum beta-lactamase (ESBL) by the combination disk method, the Vitek 1 system, and a modification of the double-disk synergy test, which is based on the inhibitory properties of clavulanic acid, tazobactam, and sulbactam. Tazobactam had the strongest effect in all seven strains. Molecular characterization of the ESBLs identified CTX-M-type enzymes, consisting of the CTX-M-9 group (n = 3), CTX-M-3 (n = 2), CTX-M-39 (n = 1), and CTX-M-2 group (n = 1). Three of the strains also carried bla-(OXA) genes and a bla-(TEM) gene. Although the prevalence of ESBLs in this study was low, further research is needed on the spread and transfer of resistance genes, both in hospitals and in the community. PMID:17265070

  14. New Delhi metallo-beta-lactamase 1-producing Enterobacteriaceae: emergence and response in Europe.

    Science.gov (United States)

    Struelens, M J; Monnet, D L; Magiorakos, A P; Santos O'Connor, F; Giesecke, J

    2010-11-18

    Acquired carbapenemases confer extensive antibiotic resistance to Enterobacteriaceae and represent a public health threat. A novel acquired carbapenemase, New Delhi metallo-beta-lactamase 1 (NDM-1), has recently been described in the United Kingdom and Sweden, mostly in patients who had received care on the Indian subcontinent. We conducted a survey among 29 European countries (the European Union Member States, Iceland and Norway) to gather information on the spread of NDM-1-producing Enterobacteriaceae in Europe, on public health responses and on available national guidance on detection, surveillance and control. A total of 77 cases were reported from 13 countries from 2008 to 2010. Klebsiella pneumoniae was the most frequently reported species with 54%. Among 55 cases with recorded travel history, 31 had previously travelled or been admitted to a hospital in India or Pakistan and five had been hospitalised in the Balkan region. Possible nosocomial acquisition accounted for 13 of 77 cases. National guidance on NDM-1 detection was available in 14 countries and on NDM-1 control in 11 countries. In conclusion, NDM-1 is spreading across Europe, where it is frequently linked to a history of healthcare abroad, but also to emerging nosocomial transmission. National guidance in response to the threat of carbapenemase-producing Enterobacteriaceae is available in approximately half of the surveyed European countries. Surveillance of carbapenemase- producing Enterobacteriaceae must be enhanced in Europe and effective control measures identified and implemented. PMID:21144431

  15. Molecular epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli

    Directory of Open Access Journals (Sweden)

    Catherine Ludden

    2014-09-01

    Full Text Available Objectives: E. coli O25b-ST131 has disseminated worldwide in hospitals and the community. The objective of this study was to determine the extent to which E. coli O25b-ST131 accounts for extended-spectrum beta-lactamase (ESBLproducing E. coli from clinical samples from all sources in this region. Methods: Between January and June 2010 ESBL-producing E. coli were collected from 94 routine samples including 47 from residents of 25 nursing homes, 15 categorized as hospital acquired and 32 others. PCR was performed for detection of bla CTX-M, bla OXA-1, bla TEM, bla SHV and for the identification of members of the E. coli O25b:ST131 clonal group. PFGE was carried out using Xba I in accordance with PulseNet protocols. Results: The majority (97% of isolates harbored a bla CTX-M gene.E. coli O25b-ST131 accounted for 87% of all ESBL-producing E. coliand for 96% of isolates from nursing home residents. Conclusion:The E. coli O25b-ST131 clonal group predominated in the collection of ESBL-producing E. coli, particularly in nursing home isolates. J Microbiol Infect Dis 2014; 4(3: 92-96

  16. Incidence of metallo-beta-lactamase-producing Pseudomonas aeruginosa in diabetes and cancer patients

    Directory of Open Access Journals (Sweden)

    Varaiya Ami

    2008-04-01

    Full Text Available Metallo-beta-lactamase (MBL-producing Pseudomonas aeruginosa strains have been reported to be an important cause of nosocomial infections. There is not enough information from India regarding their prevalence in diabetic and cancer patients. The present study was undertaken over a period of one year from January to December 2006 to study the incidence of MBL P. aeruginosa and the clinical outcome in diabetes and cancer patients admitted to S.L. Raheja Hospital, Mumbai. Two hundred and thirty isolates of P. aeruginosa were obtained from different samples of patients. These isolates were subjected to susceptibility testing to anti-pseudomonal drugs as per CLSI guidelines. They were further screened for the production of MBL by disc potentiation testing using EDTA-impregnated imipenem and meropenem discs. Of the 230 isolates of P. aeruginosa, 60 (26% isolates were found resistant to carbapenems (both imipenem and meropenem and 33 (14.3% were found to be MBL producers. Of the 33 MBL-producing isolates, 24 (72.7% were diabetic patients, six (18.1% were cancer patients and three (9% patients had both diabetes and cancer. Five (15.1% patients responded to the combination therapy of colistin, piperacillin with tazobactam and amikacin, while 28 (84.8% patients responded to the combination therapy of amikacin, piperacillin with tazobactam and gatifloxacin. Thus, the rapid dissemination of MBL producers is worrisome and necessitates the implementation of not just surveillance studies but also proper and judicious selection of antibiotics, especially carbapenems.

  17. Extended spectrum beta-lactamases in Escherichia coli from municipal wastewater

    Directory of Open Access Journals (Sweden)

    Tatiana Čornejová

    2015-09-01

    Full Text Available Introduction and objective. Over the past decades, awareness of the environmental load of resistant organisms has increased. The presented paper focuses on antibiotic resistance and detection of resistance genes in environmental [i]E. coli[/i] and on the evaluation of biofilm formation in ESBLs (extended spectrum beta-lactamase producing [i]E. coli[/i] isolated from an urban wastewater treatment plant. Materials and method. Wastewater samples and artificially added polystyrene pellets were used as the source for [i]E. col[/i]i isolation. Minimal inhibitory concentrations of 19 antibiotics were determined according to CLSI (2013. Biofilm formation was investigated by crystal violet or resazurin methods. CTX-M, carbapenemases, [i]qnrS[/i], mobile elements and virulence factors were determined by PCR. Clonal relatedness of strains was detected by principal component analysis by a Maldi biotyper. Results. ESBL phenotype was detected in 26% of environmental strains. CTX-M, CMY-2 and [i]qnrS[/i] genes of antibiotic resistance were detected. IMP gene together with integron 1 in one ertapenem resistant [i]E. coli[/i] was also recorded. There was no evident correlation between antibiotic resistance, virulence and biofilm production. Conclusions. The results showed that the wastewater is a source of ESBLs, carbapenemases and plasmid fluoroquinolone resistance. Strains with biofilm production, antibiotic resistance of CTX-M group, CMY-2, [i]qnrS[/i] genes and virulence factors present a potential environmental health risk.

  18. Beta Lactamase Producing Clostridium perfringens Bacteremia in an Elderly Man with Acute Pancreatitis

    Directory of Open Access Journals (Sweden)

    Rashmi Mishra

    2016-01-01

    Full Text Available Clostridium perfringens bacteremia is associated with adverse outcomes. Known risk factors include chronic kidney disease, malignancy, diabetes mellitus, and gastrointestinal disease. We present a 74-year-old man admitted with confusion, vomiting, and abdominal pain. Exam revealed tachycardia, hypotension, lethargy, distended abdomen, and cold extremities. He required intubation and aggressive resuscitation for septic shock. Laboratory data showed leukocytosis, metabolic acidosis, acute kidney injury, and elevated lipase. CT scan of abdomen revealed acute pancreatitis and small bowel ileus. He was started on vancomycin and piperacillin-tazobactam. Initial blood cultures were positive for C. perfringens on day five. Metronidazole and clindamycin were added to the regimen. Repeat CT (day 7 revealed pancreatic necrosis. The patient developed profound circulatory shock requiring multiple vasopressors, renal failure requiring dialysis, and bacteremia with vancomycin-resistant enterococci. Hemodynamic instability precluded surgical intervention and he succumbed to multiorgan failure. Interestingly, our isolate was beta lactamase producing. We review the epidemiology, risk factors, presentation, and management of C. perfringens bacteremia. This case indicates a need for high clinical suspicion for clostridial sepsis and that extended spectrum beta lactam antibiotic coverage may be inadequate and should be supplemented with use of clindamycin or metronidazole if culture is positive, until sensitivities are known.

  19. [Antibacterial activity and beta-lactamase stability of eleven oral cephalosporins].

    Science.gov (United States)

    Bauernfeind, A; Jungwirth, R; Schweighart, S; Theopold, M

    1990-01-01

    Oral cephalosporins (cefixime, cefdinir, cefetamet, ceftibuten, cefpodoxime, loracarbef, cefprozil, cefuroxime, cefaclor, cefadroxil and BAY 3522) were compared by their antibacterial profile including stability against new beta-lactamases. Both activity and antibacterial spectrum of compounds structurally related to third generation parenteral cephalosporins (of the oximino class) were superior to established compounds. Activity against staphylococci was found to be highest for cefdinir, cefprozil and BAY 3522. Cefetamet, ceftibuten and cefixime demonstrate no clinically meaningful antistaphylococcal activity while the other compounds investigated demonstrate intermediate activity. The antibacterial spectrum was broadest for cefdinir and cefpodoxime. New oral cephalosporins are equally inactive as established compounds against Enterobacter spp., Morganella, Listeria, Pseudomonas and Acinetobacter spp., methicillin-resistant staphylococci, Enterococcus spp., penicillin-resistant pneumococci and anaerobes. New extended broad-spectrum betalactamases (TEM-3, TEM-5, TEM-6, TEM-7, SHV-2, SHV-3, SHV-4, SHV-5, CMY-1, CMY-2, and CTX-M) are active against the majority of oral cephalosporins. Ceftibuten, cefetamet, cefixime and cefdinir were stable against some of these enzymes even to a higher extent than parenteral cephalosporins. New oral cephalosporins should improve the therapeutic perspectives of oral cephalosporins due to their higher activity against pathogens marginally susceptible to established compounds (higher multiplicity of maximum plasma concentrations over MICs of the pathogens) and furthermore by including in their spectrum organisms resistant to established absorbable cephalosporins (e.g. Proteus spp., Providencia spp., Citrobacter spp., and Serratia spp.). PMID:2079378

  20. Towards a ligand targeted enzyme prodrug therapy: single round panning of a beta-lactamase scaffold library on human cancer cells.

    Science.gov (United States)

    Shukla, Girja S; Murray, Christopher J; Estabrook, Melodie; Shen, Guang-Ping; Schellenberger, Volker; Krag, David N

    2007-05-15

    A novel beta-lactamase scaffold library in which the target-binding moiety is built into the enzyme was generated using phage display technology. The binding element is composed of a fully randomized 8 amino acid loop inserted at position between Y34 and K37 on the outer surface of Enterobacter cloacae P99 cephalosporinase (beta-lactamase, E.C. 3.5.2.6) with all library members retaining catalytic activity. The frequency and diversity of amino acids distributions in peptide inserts from library clones were analyzed. The complexity of the randomized loop appears consistent with standards of other types of phage display library systems. The library was panned against SKBR3 human breast cancer cells in 1 round using rolling circle amplification of phage DNA to recover bound phage. Individual beta-lactamase clones, independent of phage, were rapidly assessed for their binding to SKBR3 cells using a simple high throughput screen based on cell-bound beta-lactamase activity. SKBR3 cell-binding beta-lactamase enzymes were also shown to bind specifically using an immunochemical method. Selected beta-lactamase clones were further studied for their protein expression, enzyme activity and binding to nontumor cell-lines. Overall, the approach outlined here offers the opportunity of rapidly selecting targeted beta-lactamase ligands that may have a potential for their use in enzyme prodrug therapy with cephalosporin-based prodrugs. It is expected that a similar approach will be useful in developing tumor-targeting molecules of several other enzyme candidates of cancer prodrug therapy. PMID:17285581

  1. In vitro susceptibility and distribution of beta-lactamases in Enterobacteriaceae causing intra-abdominal infections in North America 2010-2011.

    Science.gov (United States)

    Hoban, Daryl J; Badal, Robert; Bouchillon, Sam; Hackel, Meredith; Kazmierczak, Krystyna; Lascols, Christine; Hawser, Stephen

    2014-07-01

    The Study for Monitoring Antimicrobial Resistance Trends has been monitoring the activity of antimicrobials indicated for the treatment of intra-abdominal infections since 2004. This report documents the in vitro activity of several recommended antimicrobials against 3449 gram-negative bacilli isolated from the 30 and 25 participating sites in North America in 2010-2011, respectively, and characterizes the extended-spectrum beta-lactamases (ESBL) identified in ESBL-positive and ertapenem-non-susceptible isolates of Enterobacteriaceae. Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Proteus mirabilis, Klebsiella oxytoca, Citrobacter freundii, Enterobacter aerogenes, Serratia marcescens, and Morganella morgannii were the most common species isolated. The incidence of beta-lactamase production was 8.8% and 8.9% for E. coli and K. pneumoniae, respectively. Overall the most active antimicrobials were amikacin, piperacillin-tazobactam, imipenem, and ertapenem, although beta-lactamase production reduced the activity of most agents. Characterization of beta-lactamase genes determined that bla(SHV), bla(CTX-M), bla(AmpC), and bla(KPC) were commonly found in most beta-lactamase-positive isolates. PMID:24813688

  2. Extended-spectrum beta-lactamase orthopedic wound infections in Nigeria

    Directory of Open Access Journals (Sweden)

    Olusolabomi J Idowu

    2011-01-01

    Full Text Available Background: Extended-spectrum beta-lactamase (ESBL-producing Gram-negative bacteria are emerging and impacting significantly on the management of patients and hospital costs. Besides, they are not being routinely sought after in diagnostic laboratories thus contributing to treatment failure. Materials and Methods: Bacterial isolates from wounds of 45 patients were identified using commercial identification kits and antibiotic susceptibility was evaluated by the Bauer-Kirby method. Screening and phenotypic confirmation of ESBL production were done as prescribed by the Clinical and Laboratory Standards Institute. The conjugation experiment was performed by the mating assay in broth between the ESBL producers and E. coli ATCC 25922 as the recipient. Results: Out of 102 Gram-negative bacteria isolated, 36 were positive for ESBL mainly of the Enterobacteriaceae family (33 and the rest were oxidase-positive bacilli (3. The predominant bacteria were Klebsiella spp. and E. coli. Others were Serratia rubidae, Citrobacter freundii, Morganella morgannii, Proteus spp., Providencia stuartii, and Enterobacter spp. There was a significant association between treatment with third-generation cephalosporins (3GCs and isolation of ESBLs ( p=0.0020 . The ESBL producers were multiply resistant and moderately sensitive to colistin. The conjugation experiment showed that the ESBL gene was transferred horizontally and tetracycline, cotrimoxazole, nitrofurantoin, gentamicin, and aztreonam resistance genes were co-transferred. No mortality was recorded but the mean length of stay in the hospital was 82 days. Conclusion: The development and spread of ESBL among Gram-negative bacteria and possible horizontal transfer calls for concern, especially in view of treatment failure, high treatment cost, and consequent discomfort to patients.

  3. A STUDY OF METALLO-BETA-LACTAMASE PRODUCING PSEUDOMONAS AERUGINOSA IN BLOOD SAMPLES OF BURNED PATIENTS

    Directory of Open Access Journals (Sweden)

    Piyali

    2014-11-01

    Full Text Available : BACKGROUND: Septicaemia is a life threatening complication of severely burned patients. Among many organisms invading blood stream Pseudomonas aeruginosa is a well-known for its powerful antibiotic resistance mechanisms which increasingly limit the choices for treatment. Among many such resistance mechanisms it is the metallo-beta-lactamase (MBL which confers resistance to Carbapenem group of antibiotics, one of the final resorts to fight them. The present study was undertaken to detect MBL producing P. aeruginosa using phenotypic method from blood samples of burned patients as well as to know their drug sensitivity pattern. MATERIALS AND METHODS: For this purpose 67 Pseudomonas aeruginosa isolates from blood samples of admitted burned patients were subjected to susceptibility testing to antipseudomonal drugs by disc diffusion test and those found to be Carbapenem resistant were subjected to Imipenem - EDTA combined disk synergy test for MBL detection. RESULT: Out of 67 isolates of P.aeruginosa, 19 (28.4% were found to be Carbapenem resistant and 11 (16.4% were MBL producers. A particularly important feature was that the MBL producers were highly resistant to the antibiotics tested than the non-producers. However all of them were susceptible to Colistin and Polymixin B. CONCLUSION: This study has made us to think that a constant vigil and careful selection of antibiotics are necessary to keep prevalence of MBL producing P.aeruginosa in check. The accurate identification and reporting of MBL producing P. aeruginosa will aid infection control practitioners in preventing the spread of these multidrug-resistant isolates

  4. Extended spectrum beta-lactamase detection in gram-negative bacilli of nosocomial origin

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    Dechen C Tsering

    2009-01-01

    Full Text Available Background: Resistance to third generation cephalosporins by acquisition and expression of extended spectrum beta lactamase (ESBL enzymes among gram-negative bacilli is on a rise. The presence of ESBL producing organisms significantly affects the course and outcome of an infection and poses a challenge to infection management worldwide. Materials and Methods: In the period from June 2007 to 2008, we collected 1489 samples from patients suspected of nosocomial infection. The isolates were identified based on colony morphology and biochemical reaction. Gram negative bacilli resistant to third generation cephalosporins were tested for ESBL by double disc synergy test (DDST- a screening test and then phenotypic confirmatory test. Antimicrobial susceptibility testing was done by modified Kirby Bauer disc diffusion method. Results: From the sample of 238 gram-negative bacilli, we isolated Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Citrobacter freundii, Proteus mirabilis, Morganella morganii and Enterobacter cloacae. Following both methods, 34% isolates were ESBL-positive. The ESBL producing isolates were significantly resistant (p < 0.01 to ampicillin, piperacillin, piperacillin/tazobactam, trimethoprim/sulfamethoxazole, tetracycline, ciprofloxacin and gentamicin as compared to non-ESBL producers. Multidrug resistance was significantly (p < 0.01 higher (69.14% in ESBL positive isolates than non-ESBL isolates (21.66%. Conclusion: High prevalence of ESBL in our hospital cannot be ignored. ESBL producers can be detected by DDST and phenotypic confirmatory test with equal efficacy. The sensitivity of screening test improved with the use of more than one antibiotic and addition of one or two antibiotics would not increase cost and labor. We recommend DDST using multiple antibiotics in all microbiology units as a routine screening test.

  5. qnrA prevalence in extended-spectrum beta-lactamase-positive Enterobacteriaceae isolates from Turkey.

    Science.gov (United States)

    Oktem, I Mehmet Ali; Gulay, Zeynep; Bicmen, Meral; Gur, Deniz

    2008-01-01

    Quinolone resistance mostly originates from chromosomal mutations. In recent years, however, plasmid-mediated quinolone resistance has been reported in several parts of the world. Plasmid-borne qnrA, qnrB, or qnrS genes are responsible for this kind of resistance. Little is known about the diversity, type, and species range of the qnr genes in Turkey. We screened qnrA, qnrB, and qnrS genes in quinolone-resistant blood culture isolates collected from six different medical centers in Turkey which produced extended-spectrum beta-lactamases (ESBLs). A total of 78 ESBL-positive isolates were enrolled in this study. Of these, 37 (47.4%) were nalidixic-acid resistant or intermediate. qnrA was found on large plasmids isolated from five (6.4%) of the Nal(I/R) isolates. In three of these, the same plasmid also carried bla(CTX-M). Four of the qnrA-positive isolates were Klebsiella pneumoniae from Dokuz Eylul University Hospital, Izmir, and the fifth isolate was Escherichia coli from Istanbul University Hospital. Two of the isolates from Izmir were found by enterobacterial repetitive interegenic consensus sequence-PCR to be clonally related. This is the first report on the qnrA prevalence among ESBL-positive blood culture isolates collected from different regions in Turkey. According to our results, plasmid-mediated resistance is a potential problem for the spread of quinolone resistance, and this mechanism could be emerging strongly among the ESBL-positive Enterobacteriaceae in Turkey. PMID:18219128

  6. Molecular modeling and docking analysis of beta-lactamases with inhibitors: a comparative study.

    Science.gov (United States)

    Danishuddin, Mohd; Khan, Asad U

    Beta-lactamases are bacterial enzymes which impart resistance against β-lactam-antibiotics. CTX-Ms are the β-lactamases that target cephalosporin antibiotics (e.g. cefotaxime and ceftazidime) while SME-1, KPC-2, IMI-1 and SFC-1 target carbapenems. Clavulanic acid, sulbactam and tazobactam are traditional β-lactamase inhibitors while LN1-255 and NXL-104 whereas novel inhibitors, inhibiting the activity of these enzymes. Studying the binding pattern of these drugs is helpful in predicting the versatile inhibitors for betalactamases. The aims of the study were: describing the mode of interaction of CTX-M (modeled from the blaCTX-M gene of this study) and the said carbapenemases with their respective target drugs and inhibitors and to perform an in silico comparison of the efficacies of traditional and novel β-lactamase-inhibitors based on fitness score. The blaCTX-M marker was PCR-amplified from plasmid DNA of E. coli strain isolated from community-acquired urinary tract infection. E. coli C600 cells (harboring cloned blaCTX-M) were found positive for extended-spectrum-β-lactamase (ESBL) production by the double-disk-synergy test. The three dimensional structures of CTX-M-15, SME-1 and IMI-1 were predicted by Swiss Model Server. The interaction between selected structures and inhibitors was performed by GOLD 5.0. On the basis of the docking score and binding pattern, we conclude that compound LN1-255 followed by tazobactam is best inhibitor against all the selected target enzymes as compared to clavulanate, sulbactam and NXL-104. Five conserved amino acids, Ser70, Ser130, Lys235, Thr236 and Gly237 were found crucial in stabilizing the complexes through hydrogen bonding and hydrophobic interactions. PMID:23202428

  7. Characterization of Beta-lactamases in Faecal Enterobacteriaceae Recovered from Healthy Humans in Spain: Focusing on AmpC Polymorphisms.

    Science.gov (United States)

    Porres-Osante, Nerea; Sáenz, Yolanda; Somalo, Sergio; Torres, Carmen

    2015-07-01

    The intestinal tract is a huge reservoir of Enterobacteriaceae, some of which are opportunist pathogens. Several genera of these bacteria harbour intrinsic antibiotic resistance genes, such as ampC genes in species of Citrobacter, Enterobacter or Escherichia genera. In this work, beta-lactamases and other resistance mechanisms have been characterized in Enterobacteriaceae isolates recovered from healthy human faecal samples, focusing on the ampC beta-lactamase genes. Fifty human faecal samples were obtained, and 70 Enterobacteriaceae bacteria were isolated: 44 Escherichia coli, 4 Citrobacter braakii, 9 Citrobacter freundii, 8 Enterobacter cloacae, 1 Proteus mirabilis, 1 Proteus vulgaris, 1 Klebsiella oxytoca, 1 Serratia sp. and 1 Cronobacter sp. A high percentage of resistance to ampicillin was detected (57%), observing the AmpC phenotype in 22 isolates (31%) and the ESBL phenotype in 3 isolates. AmpC molecular characterization showed high diversity into bla CMY and bla ACT genes from Citrobacter and Enterobacter species, respectively, and the pulsed-field gel electrophoresis (PFGE) analysis demonstrated low clonality among them. The prevalence of people colonized by strains carrying plasmid-mediated ampC genes obtained in this study was 2%. The unique plasmid-mediated bla AmpC identified in this study was the bla CMY-2 gene, detected in an E. coli isolate ascribed to the sequence type ST405 which belonged to phylogenetic group D. The hybridization and conjugation experiments demonstrated that the ISEcp1-bla CMY-2-blc structure was carried by a ~78-kb self-transferable IncK plasmid. This study shows a high polymorphism among beta-lactamase genes in Enterobacteriaceae from healthy people microbiota. Extensive AmpC-carrier studies would provide important information and could allow the anticipation of future global health problems. PMID:25501887

  8. Shedding of Clostridium difficile, fecal beta-lactamase activity, and gastrointestinal symptoms in 51 volunteers treated with oral cefixime.

    Science.gov (United States)

    Chachaty, E; Bourneix, C; Renard, S; Bonnay, M; Andremont, A

    1993-07-01

    Microbial changes including the shedding of Clostridium difficile, fecal beta-lactamase activity, and gastrointestinal symptoms were assessed in 51 healthy volunteers given 200 mg of cefixime twice daily for 8 days. The number of organisms of the family Enterobacteriaceae (means +/- standard deviations) dropped from 6.9 +/- 1.1 to 3.9 +/- 1.8 log CFU/g of feces (P < 0.01), whereas counts of enterococci rose from 7.0 +/- 1.5 to 9.0 +/- 1.0 log CFU/g of feces (P < 0.01). Both counts returned to their initial levels 50 days after the cessation of treatment. Cefixime did not significantly modify the frequency of fecal excretion of Pseudomonas aeruginosa, Staphylococcus spp., yeasts, or members of the Enterobacteriaceae resistant to ceftazidime or ampicillin. The proportion of subjects shedding C. difficile rose from 6% before treatment to 57% (P < 0.01) at the end of treatment but returned to 8% 50 days thereafter. No case of pseudomembranous colitis was observed. Stool changes occurred in 13 volunteers during treatment (25%) and in 2 others more than 10 days after the end of treatment (4%). These changes were not significantly associated with the shedding of toxigenic strains of C. difficile or with the presence of toxin A in feces. By contrast, during treatment, stool changes occurred in 8 of the 18 volunteers (44%) who had antibiotic activity in their feces but in only 5 of the 33 (15%) for whom no such activity was found (P < 0.05). The absence of antibiotic activity in the feces was itself linked with the presence of beta-lactamase activity in the feces. Since we had found earlier that fecal beta-lactamase activity afforded protection against alteration in stool consistency during treatments with oral cephalosporins, the present study confirmed our previous preliminary results in this respect. PMID:8363371

  9. Contribution of PBP3 Substitutions and TEM-1, TEM-15, and ROB-1 Beta-Lactamases to Cefotaxime Resistance in Haemophilus influenzae and Haemophilus parainfluenzae

    DEFF Research Database (Denmark)

    Søndergaard, Annette; Nørskov-Lauritsen, Niels

    2015-01-01

    AIMS: To investigate the relative contributions of naturally occurring penicillin-binding protein 3 (PBP3) substitutions, and TEM-1, TEM-15, and ROB-1 beta-lactamases on resistance to a third-generation cephalosporin in Haemophilus influenzae and Haemophilus parainfluenzae. RESULTS: The minimum...... inhibitory concentration (MIC) of cefotaxime (CTX) was assessed after transformation with PCR-amplified ftsI genes expressing altered PBP3 and/or small plasmids encoding beta-lactamases into an isogenic environment of H. influenzae and H. parainfluenzae. Group III PBP3, comprising substitutions N526K, S385T......, and L389F, conferred CTX resistance to H. influenzae according to EUCAST interpretative criteria. Group III-like PBP3, comprising substitutions N526H and S385T, increased the CTX MIC of H. parainfluenzae ninefold, but the level did not transgress the resistance breakpoint. Production of TEM-15 beta-lactamase...

  10. Rapid development in vitro and in vivo of resistance to ceftazidime in biofilm-growing Pseudomonas aeruginosa due to chromosomal beta-lactamase

    DEFF Research Database (Denmark)

    Bagge, N; Ciofu, O; Skovgaard, L T; Høiby, N

    2000-01-01

    isolated from the lungs of cystic fibrosis (CF) patients (MICceftazidime-basal/induced beta-lactamase activity: PAO 579= 0.8 mg/l-19/550 milliunits, 19676A=50 mg/l-38/957 milliunits, 17107B=100 mg/l-504/947 milliunits) were studied. After 1 or 2 weeks of continuous or intermittent (4 h/day) administration......(-1) compared to 6.0-10(-5) in the control biofilm. The same trend was observed after continuous administration of ceftazidime. MICceftazidime of the more resistant variants was increased 500-fold for PAO 579 and 8-fold for 19676A, and the specific basal beta-lactamase activities from 19 to 1,400 units for PAO......,300 units for 17107B. It was shown that, during treatment with ceftazidime, biofilm-growing P. aeruginosa had the capacity to develop resistance due to the production of chromosomal beta-lactamase....

  11. Comparison of polyacrylamide and agarose gel thin-layer isoelectric focusing for the characterization of beta-lactamases.

    OpenAIRE

    Vecoli, C; Prevost, F E; Ververis, J J; Medeiros, A A; O'Leary, G P

    1983-01-01

    Plasmid-mediated beta-lactamases from strains of Escherichia coli and Pseudomonas aeruginosa were separated by isoelectric focusing on a 0.8-mm thin-layer agarose gel with a pH gradient of 3.5 to 9.5. Their banding patterns and isoelectric points were compared with those obtained with a 2.0-mm polyacrylamide gel as the support medium. The agarose method produced banding patterns and isoelectric points which corresponded to the polyacrylamide gel data for most samples. Differences were observe...

  12. Prevalence of Extended Spectrum Beta-Lactamases among Escherichia coli and Klebsiella spp isolates in Manipal Teaching Hospital, Pokhara, Nepal

    OpenAIRE

    Shristi Raut; Shishir Gokhale; Bipin Adhikari

    2015-01-01

    Objective: To isolate, identify and phenotypically characterize extended spectrum beta-lactamase (ESBL)-producing Escherichia coli andKlebsiella spp in Manipal Teaching Hospital.Methods: Cross sectional study was conducted among E. coli and Klebsiella spp recovered from patients’ various samples to establish the prevalence of organisms producing ESBL in Manipal Teaching Hospital, Pokhara, Nepal between October 2011 and April 2012. ESBL production was detected by Clinical laboratory Standards ...

  13. Physical mapping and expression of hybrid plasmids carrying chromosomal beta-lactamase genes of Escherichia coli K-12.

    OpenAIRE

    Grundström, T; Jaurin, B; Edlund, T; Normark, S

    1980-01-01

    Hybrid plasmids carrying the ampC gene of Escherichia coli K-12 that codes for the chromosomal beta-lactamase were physically studied. The ampC gene was mapped to a deoxyribonucleic acid segment encompassing 1,370 base pairs. The mapping was facilitated by the isolation of a plasmid carrying an insertion of the transposable element gamma delta (gamma delta) close to ampC. The ampA1 mutation, which increases the expression of ampC by a factor of about 20, was localized to a 370-base pair segme...

  14. Identification of a novel ampC beta-lactamase promoter in a clinical isolate of Escherichia coli.

    OpenAIRE

    O. Olsson; Bergström, S; Normark, S

    1982-01-01

    A clinical strain of Escherichia coli, C16, that overproduces the ampC beta-lactamase was isolated. A 203-bp DNA segment from this strain, including the promoter and attenuator region of the ampC structural gene, was sequenced. A comparison with the corresponding sequence of E. coli K12 revealed four base pair differences between the ampC segments from these strains. DNA sequence data and in vitro transcription indicated that the ampC promoter in the clinical isolate was displaced 5 bp upstre...

  15. Shedding of Clostridium difficile, fecal beta-lactamase activity, and gastrointestinal symptoms in 51 volunteers treated with oral cefixime.

    OpenAIRE

    Chachaty, E; Bourneix, C; Renard, S; Bonnay, M.; Andremont, A

    1993-01-01

    Microbial changes including the shedding of Clostridium difficile, fecal beta-lactamase activity, and gastrointestinal symptoms were assessed in 51 healthy volunteers given 200 mg of cefixime twice daily for 8 days. The number of organisms of the family Enterobacteriaceae (means +/- standard deviations) dropped from 6.9 +/- 1.1 to 3.9 +/- 1.8 log CFU/g of feces (P < 0.01), whereas counts of enterococci rose from 7.0 +/- 1.5 to 9.0 +/- 1.0 log CFU/g of feces (P < 0.01). Both counts returned to...

  16. Oral treatment options for ambulatory patients with urinary tract infections caused by extended-spectrum-beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Auer, Simon; Wojna, Alexandra; Hell, Markus

    2010-09-01

    An increase in extended-spectrum-beta-lactamase (ESBL)-producing Escherichia coli has been observed in outpatient settings. Consequently, 100 ESBL-positive E. coli isolates from ambulatory patients with clinically confirmed urinary tract infections were collected by a single laboratory between October 2004 and January 2008. Antimicrobial susceptibility testing was carried out using the oral antibiotics fosfomycin, pivmecillinam, and nitrofurantoin and the parenteral antibiotic ertapenem. Susceptibility rates indicate that fosfomycin (97%), nitrofurantoin (94%), and pivmecillinam (85%) could be considered important oral treatment options. PMID:20585127

  17. Antimicrobial susceptibility and beta-lactamase production of selected gram-negative bacilli from two Croatian hospitals: MYSTIC study results.

    Science.gov (United States)

    Bedenic, B; Goic-Barisic, I; Budimir, A; Tonkic, M; Mihajkevic, L J; Novak, A; Sviben, M; Plecko, V; Punda-Polic, V; Kalenic, S

    2010-06-01

    The meropenem yearly Susceptibility Test Information Collection (MYSTIC) programme is a global, longitudinal resistance surveillance network that monitors the activity of meropenem and compares its activity with other broadspectrum antimicrobial agents. We now report the antimicrobial efficacy of meropenem compared to other broad-spectrum agents within the selective Gram-negative pathogen groups from two Croatian Hospitals investigated between 2002-2007. A total of 1510 Gram-negative pathogens were tested and the minimum-inhibitory concentrations (MICs) were determined by broth microdilution method according to CLSI.There was no resistance to either imipenem or meropenem observed for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis in both medical centers. High resistance rates of K. pneumoniae to ceftazidime (18%), cefepime (17%) and gentamicin (39%) are raising concern. Acinetobacter baumannii turned out to be the most resistant Gram-negative bacteria with 81% resistant to ceftazidime, 73% to cefepime, 69% to gentamicin and 71% to ciprofloxacin. Almost 20% of Pseudomonas aeruginosa strains were resistant to imipenem, 13% to meropenem, 69% to gentamicin and 38% to ciprofloxacin.The prevalence of extended-spectrum beta-lactamases (ESBLs) in E. coli was 10% and in K. pneumoniae 49%. PCR and sequencing of the amplicons revealed the presence of SHV-5 in nine E. coli strains and additional tem-1 beta-lactamase five strains. Five K. pneumoniae strains were positive for bla(SHV-5 )gene. Eight ESBL positive Enterobacter spp. strains were found to produce tem and CtX-m beta-lactamases. Plasmid-mediated AmpC beta-lactamases were not found among K. pneumoniae, E. coli and Enterobacter spp. Three A. baumannii strains from Zagreb University Center were identified by multiplex PCR as OXA-58 like producers. Six A. baumannii strains from Split University Center were found to possess an ISAba1 insertion sequence upstream of bla(OXA-51 )gene. According to our results

  18. Extended-spectrum beta-lactamase production among ampicillin-resistant Escherichia coli strains from chicken in Enugu State, Nigeria Produção de beta-lactamase de espectro expandido por cepas de Escherichia coli resistentes a ampicilina isoladas de frango em Enugu State, Nigéria

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    K.F. Chah

    2007-12-01

    Full Text Available One hundred and seventy-two ampicillin-resistant E. coli strains isolated from commercial chickens in Enugu State, Nigeria, were screened for beta-lactamase production using the broth method with nitrocefin® as the chromogenic cephalosporin to detect enzyme production. Beta-lactamase producing strains were further examined for extended-spectrum beta-lactamase (ESBL production using the Oxoid combination discs method. One hundred and seventy (98.8% of the 172 ampicillin-resistant E. coli strains produced beta-lactamase enzyme. Sixteen (9.4% beta-lactamase producers were phenotypically confirmed to produce ESBLs. Six of the ESBL producing strains were only detected with ceftazidime versus ceftazidime/clavulanate combination while ten of the ESBL producers were detected with cefotaxime versus cefotaxime/clavulanate combination. Chicken may serve as a reservoir of ESBL-producing E. coli strains which could be transferred to man and other animals.Cento e setenta e duas cepas de Escherichia coli resistentes a ampicilina isoladas de frangos em Enugu State, Nigéria, foram avaliadas quanto à produção de beta-lactamase através do uso de método em caldo com nitrocefin® como indicador cromogênico da produção da enzima. Em seguida, as cepas produtoras de beta-lactamase foram examinadas quanto à produção de beta-lactamase de espectro expandido (ESBL através do método de discos combinados Oxoid. Entre as cepas de Escherichia coli resistentes a ampicilina, cento e setenta (98,8% produziram beta-lactamase. Testes fenotípicos indicaram que dezesseis (9,4% das cepas produtoras de beta-lactamase produziram ESBL. Seis cepas produtoras de ESBL foram detectadas apenas com a combinação ceftazidima versus cefotaxime/clavulanato, enquanto dez cepas produtoras de ESBL foram detectadas com a combinação cefotaxime versus cefotaxime/clavulanato. Frangos podem ser reservatório de cepas de E.coli produtoras de ESBL, que podem ser transferidos para o homem

  19. Evaluation of piperacillin-tazobactam in experimental meningitis caused by a beta-lactamase-producing strain of K1-positive Escherichia coli.

    OpenAIRE

    Kern, W; Kennedy, S L; Sachdeva, M; E.R. Sande; Gunderson, D; Täuber, M G

    1990-01-01

    We evaluated the pharmacokinetics and therapeutic efficacy of piperacillin combined with tazobactam, a novel beta-lactamase inhibitor, in experimental meningitis due to a beta-lactamase-producing strain of K1-positive Escherichia coli. Different doses of piperacillin and tazobactam, as single agents and combined (8:1 ratio; dosage range, 40/5 to 200/25 mg/kg per h), and of ceftriaxone were given to experimentally infected rabbits by intravenous bolus injection followed by a 5-h constant infus...

  20. Different ratios of the piperacillin-tazobactam combination for treatment of experimental meningitis due to Klebsiella pneumoniae producing the TEM-3 extended-spectrum beta-lactamase.

    OpenAIRE

    Leleu, G; Kitzis, M D; Vallois, J M; Gutmann, L; Decazes, J M

    1994-01-01

    We evaluated the pharmacokinetics and therapeutic efficacies of piperacillin and tazobactam, a beta-lactamase inhibitor, given either alone or in different combinations (80:10, 200:10, and 80:25 mg/kg/h), in experimental meningitis due to a strain of Klebsiella pneumoniae producing the TEM-3 extended-spectrum beta-lactamase. Treatment was administered intravenously as a 7-h constant infusion preceded by a bolus of 20% of the total dose. The mean (+/- standard deviation) rates of penetration i...

  1. The translation start signal region of TEM beta-lactamase mRNA is responsible for heat shock-induced repression of amp gene expression in Escherichia coli.

    OpenAIRE

    Kuriki, Y

    1989-01-01

    pBR322 contains the amp gene encoding beta-lactamase. When Escherichia coli carrying this plasmid is exposed to heat shock, beta-lactamase synthesis is repressed transiently at the translational level. To identify the DNA element responsible for this translational repression, DNA segments containing the translation start region of the amp gene were excised from pAT153 and fused in frame with the lacZ reading frame in the open reading frame vector pORF1. These constructs were introduced into E...

  2. Activity of sulbactam in combination with ceftriaxone in vitro and in experimental endocarditis caused by Escherichia coli producing SHV-2-like beta-lactamase.

    OpenAIRE

    Fantin, B; B. Pangon; Potel, G.; Caron, F.; Vallée, E; Vallois, J M; Mohler, J.; Buré, A; Philippon, A; Carbon, C

    1990-01-01

    We studied the efficacy of sulbactam, a beta-lactamase inhibitor, in combination with ceftriaxone in vitro and in experimental endocarditis due to an Escherichia coli strain producing an extended-spectrum beta-lactamase most similar to SHV-2, a new mechanism of resistance to broad-spectrum cephalosporins among members of the family Enterobacteriaceae. In vitro, ceftriaxone demonstrated an important inoculum effect (MICs were 2 and 256 micrograms/ml with 5 X 10(5) and 5 X 10(7) CFU of inoculum...

  3. Occurrence and Detection of AmpC Beta-Lactamases among Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis Isolates at a Veterans Medical Center

    OpenAIRE

    Coudron, Philip E.; Moland, Ellen S.; Thomson, Kenneth S.

    2000-01-01

    AmpC beta-lactamases are cephalosporinases that confer resistance to a wide variety of β-lactam drugs and that may thereby create serious therapeutic problems. Although reported with increasing frequency, the true rate of occurrence of AmpC beta-lactamases in Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis remains unknown. We tested a total of 1,286 consecutive, nonrepeat isolates of these three species and found that, overall, 45 (3.5%) yielded a cefoxitin zone diameter less t...

  4. Utility of the ceftazidime-imipenem antagonism test (CIAT to detect and confirm the presence of inducible AmpC beta-lactamases among enterobacteriaceae

    Directory of Open Access Journals (Sweden)

    Vlademir Vicente Cantarelli

    2007-04-01

    Full Text Available Detection of AmpC beta-lactamase production by enterobacteria has been problematic. Contrary to ESBLs, no specific guidelines are available for detection and confirmation of AmpC production by clinical relevant microorganisms. Moreover, some bacterial species may produce inducible AmpC beta-lactamases that can be easily overlooked by routine susceptibility tests. We reported here a new test based on the strong inducible effect of imipenem on AmpC genes and the consequent antagonism with ceftazidime. This test is very simple and proved to be helpful in detecting AmpC-inducible enzymes among several species of clinical isolates.

  5. Structure-Based Design of Potent and Ligand-Efficient Inhibitors of CTX-M Class A [beta]-Lactamase

    Energy Technology Data Exchange (ETDEWEB)

    Nichols, Derek A.; Jaishankar, Priyadarshini; Larson, Wayne; Smith, Emmanuel; Liu, Guoqing; Beyrouthy, Racha; Bonnet, Richard; Renslo, Adam R.; Chen, Yu (USF); (UCSF); (Clermont)

    2012-07-11

    The emergence of CTX-M class A extended-spectrum {beta}-lactamases poses a serious health threat to the public. We have applied structure-based design to improve the potency of a novel noncovalent tetrazole-containing CTX-M inhibitor (K{sub i} = 21 {mu}M) more than 200-fold via structural modifications targeting two binding hot spots, a hydrophobic shelf formed by Pro167 and a polar site anchored by Asp240. Functional groups contacting each binding hot spot independently in initial designs were later combined to produce analogues with submicromolar potencies, including 6-trifluoromethyl-3H-benzoimidazole-4-carboxylic acid [3-(1H-tetrazol-5-yl)-phenyl]-amide, which had a K{sub i} value of 89 nM and reduced the MIC of cefotaxime by 64-fold in CTX-M-9 expressing Escherichia coli. The in vitro potency gains were accompanied by improvements in ligand efficiency (from 0.30 to 0.39) and LipE (from 1.37 to 3.86). These new analogues represent the first nM-affinity noncovalent inhibitors of a class A {beta}-lactamase. Their complex crystal structures provide valuable information about ligand binding for future inhibitor design.

  6. Multidrug resistance found in extended-spectrum beta-lactamase-producing Enterobacteriaceae from rural water reservoirs in Guantao, China

    Directory of Open Access Journals (Sweden)

    Hongna eZhang

    2015-03-01

    Full Text Available Extended-spectrum beta-lactamase (ESBL-producing Enterobacteriaceae have been isolated from humans and animals across the world. However, data on prevalence of ESBL-producing Enterobacteriaceae from rural water reservoirs is limited. This study aimed to isolate and characterize ESBL-producing Enterobacteriaceae in rural water reservoirs in Guantao, China. ESBL-producing Enterobacteriaceae were found in 5 (16.7% of 30 sampled rural water reservoirs. 66 individual isolates expressing an ESBL phenotype were obtained in the present study. Species identification showed that 42 representatives of Escherichia coli, 17 Klebsiella pneumoniae, 4 Raoultella planticola, and 3 Enterobacter cloacae. 20 isolates contained a single bla gene, including CTX-M (17 strains, TEM (2 strains, and SHV (1 strain. 46 isolates contained more than one type of beta-lactamase genes. ESBL-producing Enterobacteriaceae isolated in this study were all multidrug resistant. These findings indicated that the seroius contamination of ESBL-producing Enterobacteriaceae in rural water resevoirs existed in Guantao, China.

  7. In vivo selection of a chromosomally encoded beta-lactamase variant conferring ceftazidime resistance in Klebsiella oxytoca.

    Science.gov (United States)

    Mammeri, Hedi; Poirel, Laurent; Nordmann, Patrice

    2003-12-01

    Klebsiella oxytoca clinical isolate A was recovered from the urine of a 55-year-old man with prostatic and urinary tract infections. This isolate displayed a beta-lactam resistance phenotype consistent with overproduction of a chromosomally encoded class A beta-lactamase and had decreased susceptibilities to all beta-lactams except ceftazidime, cephamycins, and carbapenems. Four weeks after treatment with an antibiotic regimen that included ceftazidime, K. oxytoca isolate B, which had a high level of resistance to ceftazidime, was isolated from the urine of the same patient. Isoelectric focusing analysis of the culture extracts of these isolates gave a pI of 5.4 for both isolates. Cloning experiments with the PCR products of the bla(OXY) gene resulted in two Escherichia coli DH10B recombinant clones with resistance phenotypes mirroring those of the parental isolates. Sequencing analysis revealed that the bla(OXY-2-5) gene from K. oxytoca B had a single nucleotide substitution compared to the sequence of the bla(OXY-2) gene from K. oxytoca A, leading to a proline-to-serine substitution at position 167, according to the numbering of Ambler. Biochemical analysis of purified OXY-2-5 showed that it had the ability to hydrolyze ceftazidime. This is the first report of in vivo selection of a K. oxytoca isolate that produced a chromosomally encoded beta-lactamase conferring resistance to ceftazidime. PMID:14638475

  8. Mechanism of inactivation of beta-lactamases by novel 6-methylidene penems elucidated using electrospray ionization mass spectrometry.

    Science.gov (United States)

    Tabei, Keiko; Feng, Xidong; Venkatesan, Aranapakam M; Abe, Takao; Hideki, Ushirogochi; Mansour, Tarek S; Siegel, Marshall M

    2004-07-01

    The reactions of 6-methylidene penems 4-7 with beta-lactamases (TEM-1, SHV-1, Amp-C) were characterized by electrospray ionization mass spectrometry (ESI-MS). The kinetics of the reactions were monitored, demonstrating that only one penem molecule reacts to form an acyl-enzyme complex. For penem 5, the ESI-MS/MS spectrum of the hydrolysis product produced in the reaction was identical to the spectrum generated from a synthesized dihydro[1,4]thiazepine 10, confirming the rearrangement of the penem ring system to a seven-membered dihydro[1,4]thiazepine structure. Gas-phase ESI-MS/MS fragmentation data were rationalized due to tautomerization between imine and enamine substructures. ESI-MS/MS analysis of the T-6 trypsin-digested fragments of TEM-1 and SHV-1 demonstrated that the penems were only attached to Ser-70 of these class A beta-lactamases and that the penem ring structures were rearranged to seven-membered dihydro[1,4]thiazepines. PMID:15214794

  9. Metallo-beta-lactamases of Pseudomonas aeruginosa--a novel mechanism resistance to beta-lactam antibiotics.

    Directory of Open Access Journals (Sweden)

    Dorota Olszańska

    2008-06-01

    Full Text Available Since about twenty years, following the introduction into therapeutic of news beta-lactam antibiotics (broad-spectrum cephalosporins, monobactams and carbapenems, a very significant number of new beta-lactamases appeared. These enzymes confer to the bacteria which put them, the means of resisting new molecules. The genetic events involved in this evolution are of two types: evolution of old enzymes by mutation and especially appearance of new genes coming for some, from bacteria of the environment. Numerous mechanisms of enzymatic resistance to the carbapenems have been described in Pseudomonas aeruginosa. The important mechanism of inactivation carbapenems is production variety of b-lactam hydrolysing enzymes associated to carbapenemases. The metallo-beta-enzymes (IMP, VIM, SPM, GIM types are the most clinically significant carbapenemases. P. aeruginosa posses MBLs and seem to have acquired them through transmissible genetic elements (plasmids or transposons associated with integron and can be transmission to other bacteria. They have reported worldwide but mostly from South East Asia and Europe. The enzymes, belonging to the molecular class B family, are the most worrisome of all beta-lactamases because they confer resistance to carbapenems and all the beta-lactams (with the exception of aztreonam and usually to aminoglycosides and quinolones. The dissemination of MBLs genes is thought to be driven by regional consumption of extended--spectrum antibiotics (e.g. cephalosporins and carbapenems, and therefore care must be taken that these drugs are not used unnecessarily.

  10. Phenotypic and molecular characterization of extended-spectrum beta-lactamase-producing Escherichia coli in Bangladesh.

    Directory of Open Access Journals (Sweden)

    Taslima T Lina

    Full Text Available BACKGROUND: Resistance to cephalosporins in Enterobacteriaceae is mainly due to the production of extended-spectrum beta-lactamase (ESBL. Little is known about ESBL-producing bacteria in Bangladesh. Therefore, the study presents results of phenotypic and molecular characterization of ESBL-producing Escherichia coli from hospitals in Bangladesh. METHODS: A total of 339 E. coli isolated from patients with urinary tract and wound infections attending three different medical hospitals in urban and rural areas of Bangladesh between 2003-2007 were screened for ESBL-production by the double disk diffusion test. Isolates with ESBL-phenotype were further characterized by antibiotic susceptibility testing, PCR and sequencing of different β-lactamase and virulence genes, serotyping, and XbaI-macrorestriction followed by pulsed-field gel electrophoresis (PFGE. RESULTS: We identified 40 E. coli with ESBL phenotype. These isolates were resistant to ceftriaxone, ceftazidime, cefotaxime, aztreonam, cefepime, and nalidixic acid but remained susceptible to imipenem. All but one isolate were additionally resistant to ciprofloxacin, and 3 isolates were resistant to cefoxitin. ESBL genes of blaCTX-M-1-group were detected in all isolates; blaTEM-type and blaOXA-1-type genes were detected in 33 (82.5% and 19 (47.5% isolates, respectively. Virulence genes that are present in diarrhoeagenic E. coli were not found. Class-1 integron was present in 20 (50% isolates. All the ESBL-producing E. coli isolates harbored plasmids ranging between 1.1 and 120 MDa. PFGE-typing revealed 26 different pulsotypes, but identical pulsotype showed 6 isolates of serotype O25:H4. CONCLUSION: The prevalence of multidrug-resistant ESBL-producing E. coli isolates appears to be high and the majority of the isolates were positive for blaCTX-M. Although there was genetic heterogeneity among isolates, presence of a cluster of isolates belonging to serotype O25:H4 indicates dissemination of the

  11. beta-lactamase producing bacteria in the subgingival microflora of adult patients with periodontitis. A comparison between Spain and The Netherlands

    NARCIS (Netherlands)

    Herrera, D; van Winkelhoff, AJ; Dellemijn-Kippuw, N; Winkel, EG; Sanz, M

    2000-01-01

    Background/aims: Countries with a high per capita antibiotic use frequently demonstrate a high level of drug resistance. The aim of this study was to compare the prevalence and levels of beta-lactamase producing bacteria in the subgingival microflora in adult patients with periodontitis in Spain and

  12. Constitutive high expression of chromosomal beta-lactamase in Pseudomonas aeruginosa caused by a new insertion sequence (IS1669) located in ampD

    DEFF Research Database (Denmark)

    Bagge, N.; Ciofu, O.; Hentzer, Morten; Campbell, J.I.A.; Givskov, Michael Christian; Høiby, N.

    2002-01-01

    The expression of chromosomal AmpC beta-lactamase in Pseudomonas aeruginosa is negatively regulated by the activity of an amidase, AmpD. In the present study we examined resistant clinical P. aeruginosa strains and several resistant variants isolated from in vivo and in vitro biofilms for mutatio...

  13. Ceftriaxone-sulbactam combination in rabbit endocarditis caused by a strain of Klebsiella pneumoniae producing extended-broad-spectrum TEM-3 beta-lactamase.

    Science.gov (United States)

    Caron, F; Gutmann, L; Bure, A; Pangon, B; Vallois, J M; Pechinot, A; Carbon, C

    1990-11-01

    We studied the activity of the combination of sulbactam and ceftriaxone against a Klebsiella pneumoniae strain producing TEM-3, a new extended-broad-spectrum beta-lactamase, in an endocarditis model. In vitro, ceftriaxone was strongly inactivated in the presence of TEM-3 (MBC, 128 micrograms/ml with an inoculum of 5 x 10(5) CFU/ml). A marked inoculum effect was demonstrated with sulbactam: effective concentrations of inhibitor needed to reduce the MIC and MBC of ceftriaxone to similar levels increased from 1 microgram/ml in the presence of an inoculum of 5 x 10(5) CFU/ml to 20 micrograms/ml in the presence of an inoculum of 1 x 10(7) CFU/ml. In vivo, sulbactam given at 200 mg/kg of body weight every 12 h, a dosage higher than that previously reported to be effective against rabbit endocarditis caused by other microorganisms, was not sufficient to restore the complete activity of ceftriaxone given at 30 mg/kg once daily for 4 days. This insufficient activity may be correlated with the presence of a high level of beta-lactamase inside the vegetations, as indicated by a quantitative in vitro assay of beta-lactamase activity in the cardiac vegetation, suggesting an insufficient inactivation of the extended-broad-spectrum beta-lactamase in vivo. PMID:2073099

  14. Constitutive high expression of chromosomal beta-lactamase in Pseudomonas aeruginosa caused by a new insertion sequence (IS1669) located in ampD

    DEFF Research Database (Denmark)

    Bagge, Niels; Ciofu, Oana; Hentzer, Morten; Campbell, Joan I A; Givskov, Michael; Høiby, Niels

    2002-01-01

    The expression of chromosomal AmpC beta-lactamase in Pseudomonas aeruginosa is negatively regulated by the activity of an amidase, AmpD. In the present study we examined resistant clinical P. aeruginosa strains and several resistant variants isolated from in vivo and in vitro biofilms for mutations...... in the expression of high levels of AmpC beta-lactamase. Complementation of these isolates with ampD from the reference P. aeruginosa strain PAO1 caused a dramatic decrease in the expression of AmpC beta-lactamase and a parallel decrease of the MIC of ceftazidime to a level comparable to that of PAO1....... One highly resistant, constitutive beta-lactamase-producing variant contained no mutations in ampD, but a point mutation was observed in ampR, resulting in an Asp-135-->Asn change. An identical mutation of AmpR in Enterobacter cloacae has been reported to cause a 450-fold higher AmpC expression...

  15. Constitutive high expression of chromosomal beta-lactamase in Pseudomonas aeruginosa caused by a new insertion sequence (IS1669) located in ampD

    DEFF Research Database (Denmark)

    Bagge, N.; Ciofu, O.; Hentzer, Morten; Campbell, J.I.A.; Givskov, Michael Christian; Høiby, N.

    2002-01-01

    The expression of chromosomal AmpC beta-lactamase in Pseudomonas aeruginosa is negatively regulated by the activity of an amidase, AmpD. In the present study we examined resistant clinical P. aeruginosa strains and several resistant variants isolated from in vivo and in vitro biofilms for mutations...... in ampD to find evidence for the genetic changes leading to high-level expression of chromosomal beta-lactamase. A new insertion sequence, IS1669, was found located in the ampD genes of two clinical P. aeruginosa isolates and several biofilm-isolated variants. The presence of IS1669 in ampD resulted...... in the expression of high levels of AmpC beta-lactamase. Complementation of these isolates with ampD from the reference P. aeruginosa strain PAO1 caused a dramatic decrease in the expression of AmpC beta-lactamase and a parallel decrease of the MIC of ceftazidime to a level comparable to that of PAO1...

  16. Novel Insights Into The Mode of Inhibition of Class A SHV-1 Beta-Lactamases Revealed by Boronic Acid Transition State Inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    W Ke; J Sampson; C Ori; F Prati; S Drawz; C Bethel; R Bonomo; F van den Akker

    2011-12-31

    Boronic acid transition state inhibitors (BATSIs) are potent class A and C {beta}-lactamase inactivators and are of particular interest due to their reversible nature mimicking the transition state. Here, we present structural and kinetic data describing the inhibition of the SHV-1 {beta}-lactamase, a clinically important enzyme found in Klebsiella pneumoniae, by BATSI compounds possessing the R1 side chains of ceftazidime and cefoperazone and designed variants of the latter, compounds 1 and 2. The ceftazidime and cefoperazone BATSI compounds inhibit the SHV-1 {beta}-lactamase with micromolar affinity that is considerably weaker than their inhibition of other {beta}-lactamases. The solved crystal structures of these two BATSIs in complex with SHV-1 reveal a possible reason for SHV-1's relative resistance to inhibition, as the BATSIs adopt a deacylation transition state conformation compared to the usual acylation transition state conformation when complexed to other {beta}-lactamases. Active-site comparison suggests that these conformational differences might be attributed to a subtle shift of residue A237 in SHV-1. The ceftazidime BATSI structure revealed that the carboxyl-dimethyl moiety is positioned in SHV-1's carboxyl binding pocket. In contrast, the cefoperazone BATSI has its R1 group pointing away from the active site such that its phenol moiety moves residue Y105 from the active site via end-on stacking interactions. To work toward improving the affinity of the cefoperazone BATSI, we synthesized two variants in which either one or two extra carbons were added to the phenol linker. Both variants yielded improved affinity against SHV-1, possibly as a consequence of releasing the strain of its interaction with the unusual Y105 conformation.

  17. Activity of imipenem against VIM-1 metallo-beta-lactamase-producing Klebsiella pneumoniae in the murine thigh infection model.

    Science.gov (United States)

    Daikos, G L; Panagiotakopoulou, A; Tzelepi, E; Loli, A; Tzouvelekis, L S; Miriagou, V

    2007-02-01

    The in-vivo activity of imipenem against VIM-1-producing Klebsiella pneumoniae (VPKP) was assessed in a thigh infection model in neutropenic mice. Animals were infected with three VPKP isolates (imipenem MICs 2, 4 and 32 mg/L, respectively) and a susceptible clinical isolate (MIC 0.125 mg/L) that did not produce any beta-lactamase with broad-spectrum activity. Bacterial density at the site of infection was determined after imipenem treatment (30 and 60 mg/kg every 2 h for 24 h). The log(10) reduction in CFU/thigh was greatest for the wild-type isolate, intermediate for the two imipenem-susceptible VPKP isolates, and lowest for the imipenem-resistant VPKP isolate. Whilst in-vivo imipenem activity appeared reduced against in-vitro susceptible VIM-1 producers compared with a VIM-1-negative control, an increased drug dosage could moderate this reduction. PMID:17328735

  18. Spread of extended-spectrum beta-lactamase-producing Escherichia coli from a swine farm to the receiving river.

    Science.gov (United States)

    Li, Song; Song, Wengang; Zhou, Yufa; Tang, Yujing; Gao, Yanxia; Miao, Zengmin

    2015-09-01

    The dissemination of drug-resistant bacteria into different environments has posed a grave threat to public health, but data on the spread of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) from animal farms to the receiving river are limited. Here, 57 ESBL-producing E. coli isolated from a pig farm and the receiving river were analyzed in terms of drug resistance, ESBL genes, and enterobacterial repetitive intergenic consensus (ERIC). The results showed that ESBL-producing E. coli from swine feces and downstream water of the pig farm outfall overlapped substantially in drug resistance and ESBL genes. Additionally, six ESBL-producing E. coli from the downstream water exhibited 100 % genetic similarity with strains from the swine feces. In conclusion, effluents of animal farms are a likely contributor to the presence of ESBL-producing E. coli in aquatic environments. PMID:25921760

  19. Detection Of Extended-Spectrum Beta Lactamase in Klebsiella pneumoniae and Klebsiella oxytoca Bacteria with the Combined Disc Method

    Directory of Open Access Journals (Sweden)

    Ebru Yılmaz

    2011-06-01

    Full Text Available Extended-spectrum beta lactamases (ESBLs are responsible for resistance to cephalosporins (ceftazidime, ceftriaxone, and cefotaxime and aztreonam in gram-negative bacilli. ESBL producing Klebsiella bacteria are a major problem for clinicians, ESBLs increase are cause of failure in treatment particularly paediatric patients and also in medical and surgical units. In this research ESBL was investigated by combined disc method. In this research, 128 clinical isolates of Klebsiella ssp. were collected from different microbiology laboratories in Ankara. All isolates were identified with classical methods and API 20E. According to the results of identification, 103 K. pneumoniae, 25 K. oxytoca were obtained. ESBL has been detected 59,37% in Klebsiella bacteria by the combined disk method.

  20. Virulence of a Klebsiella pneumoniae strain carrying the New Delhi metallo-beta-lactamase-1 (NDM-1)

    DEFF Research Database (Denmark)

    Fuursted, Kurt; Schøler, Lone; Hansen, Frank;

    2011-01-01

    in vitro virulence by assessing various virulence factors. The NDM-1 carrying K. pneumoniae isolate was the most virulent in the murine sepsis model but there was no clear cut correlation to in vitro virulence factors or killing in C. elegans. It is concluded that K. pneumoniae carrying NDM-1 have an......The aim of the study was to compare and evaluate virulence in five strains of Klebsiella pneumoniae, including an isolate carrying New Delhi metallo-beta-lactamase-1 (NDM-1). In vivo virulence was assessed using a murine sepsis model and using the nematode Caenorhabditis elegans killing model, and...... intrinsic virulence potential, which in coexistence with its multiresistance could promote and partly explain its epidemiological success....

  1. Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus.

    Science.gov (United States)

    Aqil, Farrukh; Khan, M Sajjad A; Owais, Mohd; Ahmad, Iqbal

    2005-01-01

    Ethanolic extracts and some fractions from 10 Indian medicinal plants, known for antibacterial activity, were investigated for their ability to inhibit clinical isolates of beta-lactamase producing methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA). Synergistic interaction of plant extracts with certain antibiotics was also evaluated. The MRSA test strains were found to be multi-drug resistant and also exhibited high level of resistance to common beta-lactam antibiotics. These strains produced beta-lactamases, which hydrolyze one or other beta-lactam antibiotics, tested. The extract of the plants from Camellia sinensis (leaves), Delonix regia (flowers), Holarrhena antidysenterica (bark), Lawsonia inermis (leaves), Punica granatum (rind), Terminalia chebula (fruits) and Terminalia belerica (fruits) showed a broad-spectrum of antibacterial activity with an inhibition zone size of 11 mm to 27 mm, against all the test bacteria. The extracts from the leaves of Ocimum sanctum showed better activity against the three MRSA strains. On the other hand, extracts from Allium sativum (bulb) and Citrus sinensis (rind) exhibited little or no activity, against MRSA strains. The antibacterial potency of crude extracts was determined in terms of minimum inhibitory concentration (MIC) by the tube dilution method. MIC values, of the plant extracts, ranged from 1.3 to 8.2 mg/ml, against the test bacteria. Further, the extracts from Punica granatum and Delonix regia were fractionated in benzene, acetone and methanol. Antibacterial activity was observed in acetone as well as in the methanol fractions. In vitro synergistic interaction of crude extracts from Camellia sinensis, Lawsonia inermis, Punica granatum, Terminalia chebula and Terminalia belerica was detected with tetracycline. Moreover, the extract from Camellia sinensis also showed synergism with ampicillin.TLC of the above extracts revealed the presence of major phytocompounds, like

  2. Prevalence and antimicrobial susceptibility of extended-spectrum beta-lactamase producing urinary isolates of Escherichia coli in outpatients

    Directory of Open Access Journals (Sweden)

    Marković Tatjana

    2013-01-01

    Full Text Available Introduction. In Gram-negative bacteria, the production of beta-lactamases is the most important mechanism of resistance to beta-lactam antibiotics. In the Banja Luka region, there were no extensive researches on the prevalence and antimicrobial resistance of the extended-spectrum beta-lactamase (ESBL producing Escherichia coli (E. coli isolates. Objective. The aim of the present study was to determine the presence of ESBL producing E. coli isolates as the cause of the urinary tract infections in outpatients, the distribution of these ESBL isolates according to age and gender of patients and their susceptibility to antimicrobials. Methods. Urine specimens obtained from outpatients were cultured on chromogenic CPS-ID3 media. All plates showing significant (>105 cfu/ml growth of E. coli in pure culture were further processed. Antimicrobial susceptibility testing was performed on VITEK TWO Compact using AST-GN27 cards for testing Gram negative bacteria and detection of ESBL producers. Results. Out of 2,195 isolates, 177 (8.1% were ESBL producers. Ninety-two isolates were obtained from female patients (5% of E. coli isolated from women and 85 isolates from male patients (23% of E. coli isolated from men. High percentage of ESBL isolates was detected in the infant age group under one year (36.7% and in the age group over 60 years (28.8%. All ESBL isolates were susceptible to imipenem and resistant to ampicillin, piperacillin, cefazolin, cefotaxime, ceftazidime and cefepime. There was a significant resistance to amikacin (79.1%, gentamicin (76.8%, amoxicillin/clavulanate (54.8% and trimethoprim/sulphamethoxazole (45.8%. Resistance to nutrofurantoin was 13.6%. Conclusion. This study has demonstrated the presence of ESBL producing E. coli urinary isolates in outpatients, and their extensive susceptibility to imipenem and nitrofurantoin.

  3. Enhanced synergism of antibiotics with zinc oxide nanoparticles against extended spectrum {beta}-lactamase producers implicated in urinary tract infections

    Energy Technology Data Exchange (ETDEWEB)

    Bhande, Rashmi M., E-mail: bhanderashmi@gmail.com; Khobragade, C. N., E-mail: profcnkbt@rediffmail.com [Swami Ramanand Teerth Marathwada University, School of Life Sciences (India); Mane, R. S., E-mail: rsmane@rediffmail.com; Bhande, S., E-mail: sambhajibhande@gmail.com [Swami Ramanand Teerth Marathwada University, School of Physical Sciences (India)

    2013-01-15

    In this study, enhanced synergistic bioactivity of zinc oxide nanoparticles (ZnO NPs) with {beta}-lactam antibiotics were evaluated against a panel of clinically isolated extended spectrum {beta}-lactamase producers implicated in urinary tract infections. Chemically synthesized zinc oxide nanoparticles (15 nm) were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), high resolution transmittance electron microscopy (HR-TEM), selective area electron diffraction (SAED), X-ray photoelectron spectroscopy (XPS), and UV-Visible spectrophotometry techniques. The antimicrobial potency (10 {+-} 0.66, 12, 11.33 {+-} 1.10, and 0.7 {+-} 0.66 mm inhibiting zone) and minimum inhibitory concentrations (80, 60, 30, 50 {mu}g/ml) of ZnO NPs were tested separately whereas time-kill and membrane leakage assays were evaluated in combination with ZnO NPs+ cefotaxime, ampicillin, ceftriaxone, cefepime against the {beta}-lactamase producer strains of E. coli, K. pneumoniae, S. paucimobilis, and P. aeruginosa, respectively. Time-kill curve dynamics of ZnO NPs with {beta}-lactam antibiotics revealed enhanced bactericidal activity (50, 85, 58, 50 % fold inhibition) by delaying the exponential and stationary phases of all isolates when tested separately. Posttime-kill effect was studied on cell membrane by assaying leakage of reducing sugars (130.2, 124.7, 137, and 115.8 {mu}g/bacterial dry weight of 1 mg ({mu}g/mg) and proteins (15, 10, 16, 18 {mu}g/mg). These assays revealed that membrane leakage was due to synergism of ZnO NPs+ {beta}-lactam antibiotics which successfully damage cell membrane thereby leading to death of all ESBL producers. The results demonstrate the utilization of ZnO NPs as a potentiator of {beta}-lactam antibiotics and suggest the possibility to use nanoparticles in a combination therapy to treat UTI.

  4. Metallo-beta-lactamase producing Pseudomonas aeruginosa strains isolated in hospitals in Recife, PE, Brazil Produção de metalo-beta-lactase de linhagens de Pseudomonas aeruginosa isoladas em hospitais do Recife, PE, Brasil

    OpenAIRE

    Vera Magalhães; Ana Kelly Lins; Marcelo Magalhães

    2005-01-01

    Out of 24 nosocomial strains of Pseudomonas aeruginosa from Recife, Brazil, 15 (62%) were metallo-beta-lactamase producers. Such isolates were resistant to main antipseudomonas drugs, except polymixyn B and aztreonam. The enzyme responsible for the carbapenem-resistance belongs to SPM-1 class, and the gene involved, blaspm-1, is likely plasmid located.De 24 linhagens hospitalares de Pseudomonas aeruginosa provenientes de Recife, Brasil, 15 (62%) produziram metalo-beta-lactamase. Tais isolados...

  5. Molecular characterization of vancomycin-resistant enterococci and extended-spectrum beta-lactamase-containing Escherichia coli isolates in wild birds from the Azores Archipelago

    OpenAIRE

    2011-01-01

    Abstract To study the prevalence of vancomycin-resistant enterococci (VRE) and extended-spectrum beta-lactamases (ESBLs) containing Escherichia coli isolates, and the mechanisms of resistance implicated, 220 faecal samples from wild birds were collected between 2006 and 2010 in the Azores Archipelago. Samples were spread in Slanetz-Bartley agar plates supplemented with 4 mg/L of vancomycin and in Levine agar plates supplemented with 2 mg/L of cefotaxime for VRE and ESBL-containing ...

  6. Extensive Within-Host Diversity in Fecally Carried Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli Isolates: Implications for Transmission Analyses

    OpenAIRE

    Stoesser, N; Sheppard, A. E.; Moore, C. E.; Golubchik, T.; Parry, C M; Nget, P; Saroeun, M.; Day, N.P.; Giess, A.; Johnson, J. R.; Peto, T E; Crook, D. W.; Walker, A.S.

    2015-01-01

    Studies of the transmission epidemiology of antimicrobial-resistant Escherichia coli, such as strains harboring extended-spectrum beta-lactamase (ESBL) genes, frequently use selective culture of rectal surveillance swabs to identify isolates for molecular epidemiological investigation. Typically, only single colonies are evaluated, which risks underestimating species diversity and transmission events. We sequenced the genomes of 16 E. coli colonies from each of eight fecal samples (n = 127 ge...

  7. Characterization of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli Strains Involved in Maternal-Fetal Colonization: Prevalence of E. coli ST131

    OpenAIRE

    Birgy, André; Mariani-Kurkdjian, Patricia; Bidet, Philippe; Doit, Catherine; Genel, Nathalie; Courroux, Céline; Arlet, Guillaume; Bingen, Edouard

    2013-01-01

    Maternal-fetal Escherichia coli infections, such as neonatal bacteremia and meningitis, are important causes of morbidity and mortality. From 2006 to 2010, we studied newborns and their mothers who were colonized with E. coli in a French hospital in order to document (i) the epidemiology and genetic characteristics of extended-spectrum-beta-lactamase (ESBL)-producing E. coli strains, (ii) the prevalence of associated virulence genes, (iii) the prevalence of clone sequence type 131 (ST131), an...

  8. Ampicillin-sulbactam is effective in prevention and therapy of experimental endocarditis caused by beta-lactamase-producing coagulase-negative staphylococci.

    OpenAIRE

    Ramos, M C; Ing, M; Kim, E.; Witt, M D; Bayer, A S

    1996-01-01

    Optimal strategies for the prophylaxis and therapy of endocarditis caused by oxacillin-resistant, coagulase-negative staphylococci in patients with native or prosthetic valvular heart disease are not well defined. We compared the in vivo efficacies of ampicillin-sulbactam-based regimens with those of vancomycin-based oxacillin-resistant, beta-lactamase-producing coagulase-negative staphylococcal isolate (Staphylococcus haemolyticus SE220). Ampicillin-sulbactam (100 and 20 mg/kg of body weight...

  9. Amoxicillin treatment of experimental acute otitis media caused by Haemophilus influenzae with non-beta-lactamase-mediated resistance to beta-lactams: aspects of virulence and treatment.

    OpenAIRE

    Melhus, A; Janson, H; Westman, E.; Hermansson, A.; Forsgren, A; Prellner, K

    1997-01-01

    Through alterations primarily in the penicillin-binding proteins, a non-beta-lactamase-mediated resistance to beta-lactams has evolved in Haemophilus influenzae. The virulence of these chromosomally changed strains has been questioned. To ascertain whether these alterations involve a reduction in virulence of H. influenzae and whether they could be advantageous for the bacterium during amoxicillin treatment of acute otitis media, a total of 70 Sprague-Dawley rats were challenged with a suscep...

  10. Treatment of experimental endocarditis caused by a beta-lactamase-producing strain of Enterococcus faecalis with high-level resistance to gentamicin.

    OpenAIRE

    Hindes, R G; Willey, S H; Eliopoulos, G M; Rice, L B; Eliopoulos, C T; Murray, B.E. (Brendan); Moellering, R C

    1989-01-01

    Several antimicrobial regimens were evaluated in the treatment of experimental enterococcal endocarditis due to a beta-lactamase-producing, highly gentamicin-resistant strain of Enterococcus faecalis. Ampicillin alone cleared bacteremia in the majority of rats and reduced titers of bacteria within vegetations (6.84 versus 8.80 log10 CFU/g in controls) but did not sterilize valves. Ampicillin-sulbactam combinations, vancomycin, daptomycin, and imipenem each reduced residual bacterial titers wi...

  11. Putative connection between zoonotic multiresistant extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in dog feces from a veterinary campus and clinical isolates from dogs

    OpenAIRE

    Schaufler, Katharina; Bethe, Astrid; Lübke-Becker, Antina; Ewers, Christa; Kohn, Barbara; Wieler, Lothar H; Günther, Sebastian

    2015-01-01

    Introduction: To contribute to the understanding of multiresistant bacteria, a ‘One Health’ approach in estimating the rate of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and getting insights into the transmission from clinical settings to the surrounding environment was employed by collecting fecal samples of dogs in a public area. Isolates were compared to those from samples of diseased dogs from a nearby small-animal clinic.Materials and methods: One hundred fecal sa...

  12. Comparison of ampicillin-sulbactam with vancomycin for treatment of experimental endocarditis due to a beta-lactamase-producing, highly gentamicin-resistant isolate of Enterococcus faecalis.

    OpenAIRE

    Lavoie, S R; Wong, E S; Coudron, P E; Williams, D S; Markowitz, S M

    1993-01-01

    Increasing antibiotic resistance in the enterococci, including the capacity for beta-lactamase production and the development of high-level aminoglycoside resistance, has complicated the treatment of serious enterococcal infections, which often require synergistic antibiotic combinations for cure. We utilized the rabbit model of aortic valve endocarditis to investigate the effects of various antibiotics, alone and in combination, against a multiply antibiotic-resistant isolate of Enterococcus...

  13. Investigations of multiresistance to antibiotics and chemotherapeutics and extended spectrum beta: Lactamase effect (ESBL test) in strains E.coli and salmonella originating from domestic animals

    OpenAIRE

    Mišić Dušan; Stošić Zorica; Kiškarolj Ferenc; Adamov Vladica; Ašanin Ružica

    2006-01-01

    The presence of multiresistance to the effects of antibiotics and chemotherapeutics and extended spectrum beta-lactamase were examined in 45 strains of E. coli and 35 strains of Salmonella. The strains of E. coli originated from several species of domestic animals: dogs, cats, poultry, and cattle, and 30 strains of Salmonella originated from poultry, 4 strains from cattle, and 1 strain from swine. The presence of the following serovarieties was established using serological examinations: Salm...

  14. Effect of hyperproduction of TEM-1 beta-lactamase on in vitro susceptibility of Escherichia coli to beta-lactam antibiotics.

    OpenAIRE

    Wu, P J; Shannon, K; Phillips, I

    1994-01-01

    The susceptibility of 173 TEM-1-producing isolates of Escherichia coli was assessed by determination of MICs by the agar dilution method. MICs of amoxicillin, mezlocillin, cephaloridine, and, to a smaller extent, amoxicillin-clavulanic acid (but not cephalexin, cefuroxime, cefotaxime, ceftazidime, or imipenem) were higher for isolates that produced large amounts of beta-lactamase than for isolates that produced smaller amounts. The effect of fixed concentrations of clavulanic acid on resistan...

  15. Comparison of the inoculum effect of cefoxitin and other cephalosporins and of beta-lactamase inhibitors and their penicillin-derived components on the Bacteroides fragilis group.

    OpenAIRE

    Goldstein, E J; Citron, D M; Cherubin, C E

    1991-01-01

    We compared the inoculum effects for 109 recent clinical isolates of the Bacteroides fragilis group of cefoxitin, cefotetan, ceftizoxime, ceftriaxone, and three beta-lactamase inhibitors (clavulanic acid, sulbactam, and tazobactam) and their penicillin-derived components. Bactericidal activity was assayed and morphologic changes were monitored for selected strains exhibiting a large inoculum effect. Ceftizoxime demonstrated the largest inoculum effect, followed by cefotetan and ceftriaxone. T...

  16. TEM-4, a new plasmid-mediated beta-lactamase that hydrolyzes broad-spectrum cephalosporins in a clinical isolate of Escherichia coli.

    OpenAIRE

    Paul, G C; Gerbaud, G; Bure, A; Philippon, A M; B. Pangon; Courvalin, P.

    1989-01-01

    A clinical isolate of Escherichia coli, strain CB-134, recovered in 1986 from an abdominal abscess, exhibited resistance to penams, oxyimino-beta-lactams including broad-spectrum cephalosporins (cefotaxime, ceftriaxone, ceftazidime), and aztreonam but remained susceptible to cephamycins (cefoxitin, cefotetan) and to moxalactam and imipenem. Clavulanate (2 micrograms/ml) restored the susceptibility of the strain to broad-spectrum cephalosporins and aztreonam. A beta-lactamase with an isoelectr...

  17. Detection of extended spectrum beta-lactamases and resistance in members of the Enterobacteriaceae family isolated from healthy sheep and dogs in Umuarama, Paraná, Brazil

    OpenAIRE

    Patricia Alves de Oliveira; Rodrigo Assunção Moura; Graziela Vendrame Rodrigues; Karoline Franciani Cardoso Lopes; Melissa Marchi Zaniolo; Kariny Aparecida Jardim Rubio; Eduardo Herrera Dias; Leila Alves de Oliveira; Roberta Torres Chideroli; Daniela Dib Gonçalves

    2016-01-01

    Bacterial resistance is a primary public health concern worldwide. Within this context, pets and breeding animals act as reservoirs for multidrug-resistant bacteria (MR), such as those producing extended spectrum beta-lactamases (ESBL) and those presenting plasmid-mediated quinolone resistance (PMQR). The aim of this study was to detect the presence of ESBL and PMQR in members of the Enterobacteriaceae family, isolated from healthy sheep and dogs from non-intense farming rural properties in t...

  18. Cefotetan, a new cephamycin: comparison of in vitro antimicrobial activity with other cephems, beta-lactamase stability, and preliminary recommendations for disk diffusion testing.

    OpenAIRE

    Ayers, L W; Jones, R N; Barry, A. L.; Thornsberry, C; Fuchs, P C; Gavan, T L; Gerlach, E H; Sommers, H M

    1982-01-01

    Cefotetan is a new, potent, 7 alpha-methoxy cephalosporin (cephamycin). The in vitro activity of cefotetan tested in a multiphasic, collaborative study against 12,260 consecutive clinical isolates and 448 selected isolates showed 93% of Enterobacteriaceae, 90% of methicillin-susceptible Staphylococcus aureus (broth dilution), 83% of Bacteroides fragilis, and 72% of non-enterococcal streptococci to be inhibited by less than or equal to 8 micrograms/ml. Beta-Lactamase-producing and -nonproducin...

  19. Efficacy of ciprofloxacin in experimental aortic valve endocarditis caused by a multiply beta-lactam-resistant variant of Pseudomonas aeruginosa stably derepressed for beta-lactamase production.

    OpenAIRE

    Bayer, A S; Lindsay, P.; Yih, J; Hirano, L; Lee, D.; Blomquist, I K

    1986-01-01

    The emergence of multi-beta-lactam resistance is a limiting factor in treating invasive Pseudomonas infections with newer cephalosporins. The in vivo efficacy of ciprofloxacin, a new carboxy-quinolone, was evaluated in experimental aortic valve endocarditis caused by a strain of Pseudomonas aeruginosa which is stably derepressed for beta-lactamase production and is resistant to ceftazidime and multiple other beta-lactam agents. A total of 51 catheterized rabbits with aortic catheters in place...

  20. Study on the Genotype of Extended-spectrum Beta-lactamases Mediated by Plasmid in Southern China

    Institute of Scientific and Technical Information of China (English)

    陆坚; 唐英春; 文丽霞; 张扣兴; 张天托; 朱家馨; 谈淑卿

    2004-01-01

    To investigate the prevalence and genotype of extended spectrum beta-lactamases (ESBLs) mediated by plasmid in Gram-negative bacteria found in southern China, a total of 1184 clinical isolates of non-repetitive strains of Gram-negative bacteria were collected in 2001 from 5 different cities in southern China. The ESBLs-producing isolates were distinguished by means of the phenotype confimatory test based on the NCCLS criteria and were subjected to plasmid conjugation and electroporation experiments. Those clinical isolates succeeded in plasmid transfers had undergone plasmid conjugation and electro-transformation, plasmid DNA extraction and Pst Ⅰ digest linger-printing analysis, as well as the tmiversal primer PCR amplification of the TEM, SHV, CTX-M, VEB, PER and SFO genes and the DNA sequencing in order to determine the genotypes of ESBLs and their plasmid locations. It was found that the incidence of the ESBLs-producing strains of Gram-negative bacteria was 14.6% (173/1184) with 67 strains of transconjugants and 11 strains of electro-transformants, in which CTX-M-14 type was 33.3% (26/78); CTX-M-3 type was 23.1% (18/78); CTX-M-9 type was 14.1% (11/78); CTX-M-5 type was 6.4% (5/78); CTX-M-13 type was 2.6% (2/78); SHV-5 type was 7.7% (6/78); SHV-12 type was 5.1% (4/78), SHV-2a type was 2.6% (2/78) and unidentified type was 5.1% (4/78). 29.5% of the wild strains also carried broad-spectrum beta-lactamases TEM-1 and SHV-1 types. The above mentioned ESBLs genes were located on transferable plasmids with variable sizes (from 35 to 190 kb). The CTX-M type ESBLs was characterized by high-level of resistance to cefotaxime. It concluded that the CTX-M-type was the most prevalent genotype in clinical isolates of Gram-negative bacteria in southern China, and the SHVtype ranks in the second place. TEM-, VEB-, Toho- and PER-types were not found in these isolates.

  1. [Typing of extended-spectrum beta-lactamase-producing Salmonella typhimurium strains isolated in a pediatric unit].

    Science.gov (United States)

    Mhand, R A; Soukri, A; Amarouch, H; Mdaghri, N E; Benbachir, M

    1999-01-01

    Extended-spectrum b-lactamases (ESBLs) derive mainly from TEM and SHV b-lactamases. These enzymes confer resistance to all oxyimino cephalosporins and monobactams except cephamycins and carbapems. ESBLs are often encoded by large plasmids that carry resistance determinants to multiple antibiotics and spread among the members of the Enterobacteriaceae. Since the first outbreak of Klebsiella pneumoniae expressing an extended-spectrum beta-lactamase reported in 1984, nosocomial infections due to Enterobacteriaceae species which produce ESBLs have been generally recovered from patients hospitalized in intensive care units. The most frequently isolated ESBL-producing strains belong to the genus Klebsiella, Escherichia, Enterobacter and Proteus; ESBLs are rarely associated with the genus Salmonella. The first Salmonella were detected in France in 1984 (Salmonella typhimurium), in Tunisia in 1988 (Salmonella wien) and in Argentina in 1991 (Salmonella typhimurium). In 1994, 10 isolates of Salmonella typhimurium expressing an extended-spectrum beta-lactamase were isolated for the first time from 10 children hospitalized in a pediatric unit of the hospital Ibn-Rochd, Casablanca. Previous study showed that all isolates belonged the same serotype, and biotype, and showed a resistance to oxyimino beta-lactams, gentamycin, tobramycin and trimethoprim-sulfamethoxazole but remained susceptible to tetracycline, chloramphenicol and quinolones. Oxyimino beta-lactams resistance determinant of all strains of Salmonella typhimurium was transferred by conjugation to Escherichia coli; Resistance to gentamycin and trimethoprim-sulfamethoxazole was also cotransferred. In this study, we characterized the relationship between all isolates by comparing plasmid profiles and patterns of proteins because there appear to be the more effective method for evaluating epidemiologic relationship between Salmonella species, and the protein profiles method has been used for many bacterial species. These

  2. The Structural Bases of Antibiotic Resistance in the Clinically Derived Mutant beta-Lactamases TEM-30, TEM-32, and TEM-34

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Xiaojun; Minasov, George; Shoichet, Brian K. (NWU)

    2010-03-08

    Widespread use of {beta}-lactam antibiotics has promoted the evolution of {beta}-lactamase mutant enzymes that can hydrolyze ever newer classes of these drugs. Among the most pernicious mutants are the inhibitor-resistant TEM {beta}-lactamases (IRTs), which elude mechanism-based inhibitors, such as clavulanate. Despite much research on these IRTs, little is known about the structural bases of their action. This has made it difficult to understand how many of the resistance substitutions act as they often occur far from Ser-130. Here, three IRT structures, TEM-30 (R244S), TEM-32 (M69I/M182T), and TEM-34 (M69V), are determined by x-ray crystallography at 2.00, 1.61, and 1.52 {angstrom}, respectively. In TEM-30, the Arg-244 {yields} Ser substitution (7.8 {angstrom} from Ser-130) displaces a conserved water molecule that usually interacts with the {beta}-lactam C3 carboxylate. In TEM-32, the substitution Met-69 {yields} Ile (10 {angstrom} from Ser-130) appears to distort Ser-70, which in turn causes Ser-130 to adopt a new conformation, moving its O{gamma} further away, 2.3 {angstrom} from where the inhibitor would bind. This substitution also destabilizes the enzyme by 1.3 kcal/mol. The Met-182 {yields} Thr substitution (20 {angstrom} from Ser-130) has no effect on enzyme activity but rather restabilizes the enzyme by 2.9 kcal/mol. In TEM-34, the Met-69 {yields} Val substitution similarly leads to a conformational change in Ser-130, this time causing it to hydrogen bond with Lys-73 and Lys-234. This masks the lone pair electrons of Ser-130 O{gamma}, reducing its nucleophilicity for cross-linking. In these three structures, distant substitutions result in accommodations that converge on the same point of action, the local environment of Ser-130. TEM-1 {beta}-lactamase is the predominant source of resistance to {beta}-lactams, such as the penicillins. TEM-1 and related class A {beta}-lactamases confer resistance by hydrolyzing the {beta}-lactam ring of these antibiotics

  3. Characterization of VIM-2, a carbapenem-hydrolyzing metallo-beta-lactamase and its plasmid- and integron-borne gene from a Pseudomonas aeruginosa clinical isolate in France.

    Science.gov (United States)

    Poirel, L; Naas, T; Nicolas, D; Collet, L; Bellais, S; Cavallo, J D; Nordmann, P

    2000-04-01

    Pseudomonas aeruginosa COL-1 was identified in a blood culture of a 39-year-old-woman treated with imipenem in Marseilles, France, in 1996. This strain was resistant to beta-lactams, including ureidopenicillins, ticarcillin-clavulanic acid, cefepime, ceftazidime, imipenem, and meropenem, but remained susceptible to the monobactam aztreonam. The carbapenem-hydrolyzing beta-lactamase gene of P. aeruginosa COL-1 was cloned, sequenced, and expressed in Escherichia coli DH10B. The deduced 266-amino-acid protein was an Ambler class B beta-lactamase, with amino acid identities of 32% with B-II from Bacillus cereus; 31% with IMP-1 from several gram-negative rods in Japan, including P. aeruginosa; 27% with CcrA from Bacteroides fragilis; 24% with BlaB from Chryseobacterium meningosepticum; 24% with IND-1 from Chryseobacterium indologenes; 21% with CphA-1 from Aeromonas hydrophila; and 11% with L-1 from Stenotrophomonas maltophilia. It was most closely related to VIM-1 beta-lactamase recently reported from Italian P. aeruginosa clinical isolates (90% amino acid identity). Purified VIM-2 beta-lactamase had a pI of 5.6, a relative molecular mass of 29.7 kDa, and a broad substrate hydrolysis range, including penicillins, cephalosporins, cephamycins, oxacephamycins, and carbapenems, but not monobactams. As a metallo-beta-lactamase, its activity was zinc dependent and inhibited by EDTA (50% inhibitory concentration, 50 microM). VIM-2 conferred a resistance pattern to beta-lactams in E. coli DH10B that paralleled its in vitro hydrolytic properties, except for susceptibility to ureidopenicillins, carbapenems, and cefepime. bla(VIM-2) was located on a ca. 45-kb plasmid that in addition conferred resistance to sulfamides and that was not self-transmissible either from P. aeruginosa to E. coli or from E. coli to E. coli. bla(VIM-2) was the only gene cassette located within the variable region of a novel class 1 integron, In56, that was weakly related to the bla(VIM-1)-containing

  4. Sequential necrotizing fasciitis caused by the monomicrobial pathogens Streptococcus equisimilis and extended-spectrum beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Endo, Akiko; Matsuoka, Ryosuke; Mizuno, Yasushi; Doi, Asako; Nishioka, Hiroaki

    2016-08-01

    Necrotizing fasciitis is a rapidly progressing bacterial infection of the superficial fascia and subcutaneous tissue that is associated with a high mortality rate and is caused by a single species of bacteria or polymicrobial organisms. Escherichia coli is rarely isolated from patients with monomicrobial disease. Further, there are few reports of extended-spectrum beta-lactamase (ESBL)-producing E. coli associated with necrotizing fasciitis. We report here our treatment of an 85-year-old man who was admitted because of necrotizing fasciitis of his right thigh. Streptococcus equisimilis was detected as a monomicrobial pathogen, and the infection was cured by amputation of the patient's right leg and the administration of antibiotics. However, 5 days after discontinuing antibiotic therapy, he developed necrotizing fasciitis on his right upper limb and died. ESBL-producing E. coli was the only bacterial species isolated from blood and skin cultures. This case demonstrates that ESBL-producing E. coli can cause monomicrobial necrotizing fasciitis, particularly during hospitalization and that a different bacterial species can cause disease shortly after a previous episode. PMID:26912298

  5. Instant Typing Is Essential to Detect Transmission of Extended-Spectrum Beta-Lactamase-Producing Klebsiella Species.

    Directory of Open Access Journals (Sweden)

    Anne F Voor in 't Holt

    Full Text Available Infections with multidrug-resistant (MDR microorganisms are an increasing threat to hospitalized patients. Although rapid typing of MDR microorganisms is required to apply targeted prevention measures, technical barriers often prevent this. We aimed to assess whether extended-spectrum beta-lactamase (ESBL-producing Klebsiella species are transmitted between patients and whether routine, rapid typing is needed.For 43 months, the clonality of all ESBL-producing Klebsiella isolates from patients admitted to Erasmus MC University Medical Center in Rotterdam, the Netherlands was assessed with Raman spectroscopy. A cluster was defined as n ≥ 2 patients who had identical isolates. Primary patients were the first patients in each cluster. Secondary patients were those identified with an isolate clonally related to the isolate of the primary patient.Isolates from 132 patients were analyzed. We identified 17 clusters, with 17 primary and 56 secondary patients. Fifty-nine patients had a unique isolate. Patients (n = 15 in four out of the 17 clusters were epidemiologically related. Ten of these 15 patients developed an infection.Clonal outbreaks of ESBL-producing Klebsiella species were detected in our hospital. Theoretically, after Raman spectroscopy had detected a cluster of n ≥ 2, six infections in secondary patients could have been prevented. These findings demonstrate that spread of ESBL-producing Klebsiella species occurs, even in a non-outbreak setting, and underscore the need for routine rapid typing of these MDR bacteria.

  6. Extended spectrum beta-lactamase-producing Enterobacteriaceae in international travelers and non-travelers in New York City.

    Directory of Open Access Journals (Sweden)

    Scott A Weisenberg

    Full Text Available BACKGROUND: We performed this study 1 to determine the prevalence of community-associated extended spectrum beta-lactamase producing Enterobacteriaceae (ESBLPE colonization and infection in New York City (NYC; 2 to determine the prevalence of newly-acquired ESBLPE during travel; 3 to look for similarities in contemporaneous hospital-associated bloodstream ESBLPE and travel-associated ESBLPE. METHODS: Subjects were recruited from a travel medicine practice and consented to submit pre- and post-travel stools, which were assessed for the presence of ESBLPE. Pre-travel stools and stools submitted for culture were used to estimate the prevalence of community-associated ESBLPE. The prevalence of ESBLPE-associated urinary tract infections was calculated from available retrospective data. Hospital-associated ESBLPE were acquired from saved bloodstream isolates. All ESBLPE underwent multilocus sequence typing (MLST and ESBL characterization. RESULTS: One of 60 (1.7% pre- or non-travel associated stool was colonized with ESBLPE. Among community-associated urine specimens, 1.3% of Escherichia coli and 1.4% of Klebsiella pneumoniae were identified as ESBLPE. Seven of 28 travelers (25.0% acquired a new ESBLPE during travel. No similarities were found between travel-associated ESBLPE and hospital-associated ESBLPE. A range of imported ESBL genes were found, including CTX-M-14 and CTX-15. CONCLUSION: ESBL colonization and infection were relatively low during the study period in NYC. A significant minority of travelers acquired new ESBLPE during travel.

  7. Comparison between phenotypic and PCR for detection of OXA-23 type and metallo-beta-lactamases producer Acinetobacter spp.

    Directory of Open Access Journals (Sweden)

    Azimi, Leila

    2013-11-01

    Full Text Available [english] Background: Resistance to carbapenems is developing around the world and can cause many problems for treatment of patients. Production of metallo-beta-lactamase (MBL is one of the main mechanism for this type of resistance. So, detection of MBL-producer microorganisms can prevent the spread of this type of resistance.Materials and methods: In this study 94 spp. were investigated. Resistance to imipenem was conducted after purification and identification. Combination disc (CD and Double Disc Synergy Test (DDST were performed for phenotypic detection of MBL and the molecular PCR method was done for vim-1, vim-2, imp-1 and OXA-23 genes.Results: According to TSI, SIM and oxidation-fermentation (OF test and PCR assay 93 and one strain were identified. 85% of them were resistant to imipenem. 34% of them have a positive combination disc test (CD while Double Disc Synergy Test (DDST was negative for all of them. The vim-1, vim-2 and imp-1 genes were not detected in PCR molecular method, however in 74% of strains with positive results in combination disc, were positive for the OXA-23 gene after PCR test. This study shows that the blaOXA-23 resistance determinant may become an emerging therapeutic problem.Discussion: According to the results, it seems that combination disc does not have enough specificity for detection of MBL-producer and using Double Disc Synergy Test (DDST can be more convenient.

  8. Clavulanic acid: a competitive inhibitor of beta-lactamases with novel anxiolytic-like activity and minimal side effects.

    Science.gov (United States)

    Kim, Deog J; King, Jean A; Zuccarelli, Lisa; Ferris, Craig F; Koppel, Gary A; Snowdon, Charles T; Ahn, Chang H

    2009-08-01

    Clavulanic acid is a member of the beta lactam family of antibiotics with little or no intrinsic antibacterial activity of its own; instead, it is used to enhance the activity of antibiotics by blocking bacterial beta-lactamases. Because clavulanic acid by itself is very safe, orally active and shows good brain penetrance, we sought to determine if it had any potential as a psychotherapeutic. Clavulanic acid was a tested across three mammalian species, hamsters, rats and cotton-top tamarin monkeys in a series of behavioral assays designed to screen for anxiolytic activity. In addition, several studies were done in rodents to compare the behavioral profile of clavulanic acid to the commonly prescribed benzodiazepines, particularly with respect to their unwanted side effects of motor depression, amnesia and neuroendocrine dysregulation. Our findings show that clavulanic acid is a highly potent anxiolytic in rodents without altering motor activity in the open field test, normal learning and memory in the Morris water maze, or normal stress hormone release. Orally administered clavulanic acid significantly reduces measures of anxiety in male/female pairs of cotton-top tamarins. In addition, male tamarins showed a highly significant increase in sexual arousal as measured by the number of penile erections. The fact clavulanic acid has anxiolytic activity in the tamarin holds the promise that this drug may be an effective therapeutic for the treatment of anxiety disorders in humans. PMID:19394358

  9. Phenotypic Detection of Metallo-Beta-Lactamases in Carbapenem Resistant Acinetobacter baumannii Isolated from Pediatric Patients in Pakistan

    Directory of Open Access Journals (Sweden)

    Muneeza Anwar

    2016-01-01

    Full Text Available Multidrug resistant A. baumannii has emerged as an important and problematic human pathogen as it is the causative agent of several types of infections especially in neonates and immunocompromised patients because they have least capacity to fight against infections. Carbapenems are used as last resort antibiotics for treating these infections but currently resistance against carbapenems due to MBL production is on the rise. The objective of this study was to determine the frequency of antibiotic resistance in A. baumannii and also to compare the efficacy of combined disk test and double disk synergy test for detection of metallo-beta-lactamases. A total of 112 A. baumannii were identified from various clinical samples and antibiotic susceptibility profile was determined by Kirby-Bauer Disk Diffusion method. Out of 112, 66 (58.9% isolates were resistant to both imipenem and meropenem (OXOID. These resistant isolates were tested for carbapenemase production, and 55 (83.3% were carbapenemase producers by Modified Hodge Test. These isolates were further tested for MBL production by combined disk test and double disk synergy test. Out of 66, 49 isolates were positive by both methods, CDT and DDST, and only one isolate was detected as negative (with kappa value = 0.038. All MBL producing strains showed remarkable resistance to cephalosporins, fluoroquinolones, aminoglycosides, and piperacillin/tazobactam (OXOID. The antibiotic resistance was very high in A. baumannii which were isolated from children in Pakistan specially attending a nephrology unit.

  10. Phenotypic Detection of Metallo-Beta-Lactamases in Carbapenem Resistant Acinetobacter baumannii Isolated from Pediatric Patients in Pakistan

    Science.gov (United States)

    Anwar, Muneeza; Ejaz, Hassan; Zafar, Aizza; Hamid, Hamdan

    2016-01-01

    Multidrug resistant A. baumannii has emerged as an important and problematic human pathogen as it is the causative agent of several types of infections especially in neonates and immunocompromised patients because they have least capacity to fight against infections. Carbapenems are used as last resort antibiotics for treating these infections but currently resistance against carbapenems due to MBL production is on the rise. The objective of this study was to determine the frequency of antibiotic resistance in A. baumannii and also to compare the efficacy of combined disk test and double disk synergy test for detection of metallo-beta-lactamases. A total of 112 A. baumannii were identified from various clinical samples and antibiotic susceptibility profile was determined by Kirby-Bauer Disk Diffusion method. Out of 112, 66 (58.9%) isolates were resistant to both imipenem and meropenem (OXOID). These resistant isolates were tested for carbapenemase production, and 55 (83.3%) were carbapenemase producers by Modified Hodge Test. These isolates were further tested for MBL production by combined disk test and double disk synergy test. Out of 66, 49 isolates were positive by both methods, CDT and DDST, and only one isolate was detected as negative (with kappa value = 0.038). All MBL producing strains showed remarkable resistance to cephalosporins, fluoroquinolones, aminoglycosides, and piperacillin/tazobactam (OXOID). The antibiotic resistance was very high in A. baumannii which were isolated from children in Pakistan specially attending a nephrology unit. PMID:27123345

  11. Antibiotic combinatorial approach utilized against extended spectrum beta-lactamase (ESBL bacteria isolates from Enugu, South Eastern Nigeria

    Directory of Open Access Journals (Sweden)

    Ruth A. Afunwa

    2014-04-01

    Full Text Available Introduction: Antibiotic options in the treatment of extended spectrum beta-lactamase (ESBL producing bacteria are very limited. The purpose of this study was to analyze several commonly applied antibiotics in quite various novel combinations for use against ESBL-producing bacteria isolates.Methods: Total of 460 samples of urine, throat and anal swab were collected from volunteers and patients from nursery, primary and secondary schools and from other individuals in the community. Hospital and community isolates comprised of 65% and 35% respectively. The identification and characterization of the isolates were done by standard culturing and in vitro antibiotic sensitivity procedures.Results: The antibiotic combination studies showed that the combination of gentamicin with the other antibiotics had predominantly synergistic effects. The percentage synergistic effect for the combinations of gentamicin/pefloxacin was 69%, gentamicin/[Amoxicillin and clavulanic acid] 72%, gentamicin/ceftriaxone 68%, gentamicin/cefuroxime 81.9%, and gentamicin/ciprofloxacin 80.6%, against the community and hospital derived ESBL producing organisms of both Enterobacteriaceae and Pseudomonas species.Conclusion: Good antimicrobial monitoring exercise and corresponding antimicrobial screening activities should work towards a dynamic approach to generate effective treatment options using combination therapy.

  12. Coexistence of plasmid-mediated quinolone resistance determinants and AmpC-Beta-Lactamases in Escherichia coli strains in Egypt.

    Science.gov (United States)

    Abd El-Aziz, N K; Gharib, A A

    2015-01-01

    Three kinds of plasmid—mediated quinolone resistance (PMQR) determinants (qnr genes, qepA and aac(6')—Ib—cr) have been discovered and shown to be widely distributed among clinical isolates. To characterize the prevalence of PMQR determinants among AmpC—producing E. coli strains in food—producing animals and animal by—products in Egypt, twenty—nine E. coli strains were tested for their susceptibilities to antimicrobials and screened for PMQR determinants and AmpC Beta lactamases using PCR and plasmid profiling. It was found that qnr genes being detected alone or in combination with qepA or aac(6')—Ib—cr genes in 11 (37.9%) strains comprising 9 for qnrA and only one for both qnrB and qnrS. Moreover, qepA and aac(6')—Ib—cr were detected in 41.38% and 3.45% of E. coli strains, respectively. The ampC β—lactamase genes were detected in 75.86 % of all strains and in 100% and 53.3% of the PMQR determinant—positive and negative strains, respectively. In several cases, plasmid profiling of E. coli strains exhibiting the coexistence of both PMQR determinants and ampC genes on a single plasmid as a first report in Egypt that may contribute to rapid spread and increase in bacterial resistance, which is important to public health concern. PMID:26475385

  13. High diversity of extended-spectrum beta-lactamase-producing bacteria in an urban river sediment habitat.

    Science.gov (United States)

    Lu, Su-Ying; Zhang, Ya-Li; Geng, Sui-Na; Li, Tian-Yu; Ye, Zhuo-Ming; Zhang, Dong-Sheng; Zou, Fei; Zhou, Hong-Wei

    2010-09-01

    Antibiotic-resistant bacteria (ARB) have been surveyed widely in water bodies, but few studies have determined the diversity of ARB in sediment, which is the most taxon-abundant habitat in aquatic environments. We isolated 56 extended-spectrum beta-lactamase (ESBL)-producing bacteria from a single sediment sample taken from an urban river in China. All strains were confirmed for ESBL-producing capability by both the clavulanic acid combination disc method and MIC determination. Of the isolated strains, 39 were classified as Enterobacteriaceae (consisting of the genera Escherichia, Klebsiella, Serratia, and Aeromonas) by 16S rRNA gene sequencing and biochemical analysis. The present study identifies, for the first time, ESBL-producing strains from the families Brucellaceae and Moraxellaceae. The bla(CTX-M) gene was the most dominant of the ESBL genes (45 strains), while the bla(TEM) gene was the second-most dominant (22 strains). A total of five types of bla(CTX-M) fragments were identified, with both known and novel sequences. A library of bla(CTX-M) cloned from the sediment DNA showed an even higher diversity of bla(CTX-M) sequences. The discovery of highly diverse ESBL-producing bacteria and ESBL genes, particularly bla(CTX), in urban river sediment raises alarms for potential dissemination of ARB in communities through river environments. PMID:20639374

  14. The KPC type beta-lactamases: new enzymes that confer resistance to carbapenems in Gram-negative bacilli.

    Directory of Open Access Journals (Sweden)

    Piotr Wieczorek

    2010-05-01

    Full Text Available Antimicrobial resistance due to the continuous selective pressure from widespread use of antimicrobials in humans, animals and agriculture has been a growing problem for last decades. KPC beta-lactamases hydrolyzed beta-lactams of all classes. Especially, carbapenem antibiotics are hydrolyzed more efficiency than other beta-lactam antibiotics. The KPC enzymes are found most often in Enterobacteriaceae. Recently, these enzymes have been found in isolates of Pseudomonas aeruginosa and Acinetobacter spp. The observations of blaKPC genes isolated from different species in other countries indicate that these genes from common but unknown ancestor may have been mobilized in these areas or that blaKPC-carrying bacteria may have been passively by many vectors. The emergence of carbapenem resistance in Gram-negative bacteria is worrisome because the carbapenem resistance often may be associated with resistance to many beta-lactam and non-beta-lactam antibiotics. Treatment of infections caused by KPC-producing bacteria is extremely difficult because of their multidrug resistance, which results in high mortality rates. Therapeutic options to treat infections caused by multiresistant Gram-negative bacteria producing KPC-carbapenemases could be used polymyxin B or tigecycline.

  15. Differences in Extended-Spectrum Beta-Lactamase Producing Escherichia coli Virulence Factor Genes in the Baltic Sea Region

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    Jana Lillo

    2014-01-01

    Full Text Available The aim of this study was to compare the prevalence of different virulence factor (VF genes in extended-spectrum beta-lactamase (ESBL producing Escherichia coli strains isolated from the Baltic Sea region. A total of 432 strains of phenotypically ESBL positive E. coli were collected from 20 institutions located in Estonia, Latvia, Lithuania, and the region of St. Petersburg in Russia from January to May 2012 and analyzed for phylogenetic group and prevalence of 23 VF genes. The strains were collected from clinical material (urine, blood, wound, and respiratory tract. Bacterial isolates were compared according to phylogenetic group, clinical material, and geographical origin. Most of the VF genes were concentrated within phylogenetic group B2 and/or D. When comparing strains isolated from different countries, it was found that strains originating from Estonia and Latvia belonged mainly to group B2 and strains from Lithuania and Russia mainly to groups B2 and D. The P-fimbrial adhesin gene papEF was more prevalent in Russian strains, colicin gene cvaC in Lithuanian strains, and capsular gene kpsMTII in Latvian strains; serum resistant gene traT was less prevalent in Estonian strains. The regional differences of VF genes remained statistically significant after taking into account the phylogenetic distribution in the countries.

  16. Differences in extended-spectrum beta-lactamase producing Escherichia coli virulence factor genes in the Baltic Sea region.

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    Lillo, Jana; Pai, Kristiine; Balode, Arta; Makarova, Mariia; Huik, Kristi; Kõljalg, Siiri; Ivanova, Marina; Kaftyreva, Lidia; Miciuleviciene, Jolanta; Naaber, Paul; Parv, Kristel; Pavelkovich, Anastasia; Rööp, Tiiu; Toompere, Karolin; Suzhaeva, Ludmila; Sepp, Epp

    2014-01-01

    The aim of this study was to compare the prevalence of different virulence factor (VF) genes in extended-spectrum beta-lactamase (ESBL) producing Escherichia coli strains isolated from the Baltic Sea region. A total of 432 strains of phenotypically ESBL positive E. coli were collected from 20 institutions located in Estonia, Latvia, Lithuania, and the region of St. Petersburg in Russia from January to May 2012 and analyzed for phylogenetic group and prevalence of 23 VF genes. The strains were collected from clinical material (urine, blood, wound, and respiratory tract). Bacterial isolates were compared according to phylogenetic group, clinical material, and geographical origin. Most of the VF genes were concentrated within phylogenetic group B2 and/or D. When comparing strains isolated from different countries, it was found that strains originating from Estonia and Latvia belonged mainly to group B2 and strains from Lithuania and Russia mainly to groups B2 and D. The P-fimbrial adhesin gene papEF was more prevalent in Russian strains, colicin gene cvaC in Lithuanian strains, and capsular gene kpsMTII in Latvian strains; serum resistant gene traT was less prevalent in Estonian strains. The regional differences of VF genes remained statistically significant after taking into account the phylogenetic distribution in the countries. PMID:25250320

  17. Differences in Extended-Spectrum Beta-Lactamase Producing Escherichia coli Virulence Factor Genes in the Baltic Sea Region

    Science.gov (United States)

    Balode, Arta; Makarova, Mariia; Huik, Kristi; Kõljalg, Siiri; Kaftyreva, Lidia; Miciuleviciene, Jolanta; Naaber, Paul; Rööp, Tiiu; Toompere, Karolin; Suzhaeva, Ludmila; Sepp, Epp

    2014-01-01

    The aim of this study was to compare the prevalence of different virulence factor (VF) genes in extended-spectrum beta-lactamase (ESBL) producing Escherichia coli strains isolated from the Baltic Sea region. A total of 432 strains of phenotypically ESBL positive E. coli were collected from 20 institutions located in Estonia, Latvia, Lithuania, and the region of St. Petersburg in Russia from January to May 2012 and analyzed for phylogenetic group and prevalence of 23 VF genes. The strains were collected from clinical material (urine, blood, wound, and respiratory tract). Bacterial isolates were compared according to phylogenetic group, clinical material, and geographical origin. Most of the VF genes were concentrated within phylogenetic group B2 and/or D. When comparing strains isolated from different countries, it was found that strains originating from Estonia and Latvia belonged mainly to group B2 and strains from Lithuania and Russia mainly to groups B2 and D. The P-fimbrial adhesin gene papEF was more prevalent in Russian strains, colicin gene cvaC in Lithuanian strains, and capsular gene kpsMTII in Latvian strains; serum resistant gene traT was less prevalent in Estonian strains. The regional differences of VF genes remained statistically significant after taking into account the phylogenetic distribution in the countries. PMID:25250320

  18. Occurrence of extended-spectrum beta-lactamases in Enterobacteriaceae isolated from hospitalized patients in Curitiba, southern Brazil

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    Keite da Silva Nogueira

    2006-12-01

    Full Text Available Production of extended-spectrum beta-lactamases (ESBL by enterobacteria is an important resistance mechanism against antimicrobial beta-lactamics. We tested 498 bacterial strains isolated from two tertiary-care teaching hospitals for ESBL production, using screening breakpoints for aztreonam and third generation cephalosporins, according to CLSI recommendations. Among these isolates, 155 were positive for the ESBL screening test, and 121 (78% were confirmed by the clavulanic acid combination disk method. We found a high frequency of ESBL (24% among Enterobacteriaceae, with a frequency of 57.4% for Klebsiella pneumoniae, 21.4% for Klebsiella oxytoca, and 7.2% for E. coli. In other members of Enterobacteriaceae, non-Klebsiella and non-E. coli, the prevalence was 21.6%. Ceftriaxone and cefotaxime showed a higher sensitivity in the screening test (99.2% when compared to ceftazidime, aztreonam and cefpodoxime. However, cefotaxime/cefotaxime plus clavulanic acid showed a higher sensitivity in the confirmatory test (96.7%.

  19. High prevalence rate of extended-spectrum beta-lactamases (ESBL among Enterobacteriaceae in a small Brazilian public hospital

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    Adriane Lenhard-Vidal

    2011-12-01

    Full Text Available The production of extended-spectrum beta-lactamases (ESBL is considered one of the most important resistance mechanisms that impair antimicrobial treatment of infections caused by Enterobacteriaceae. Data on culture and susceptibility tests were collected from the Clinical Analyses and Research Laboratory charts reporting on patients admitted to the University Hospital of Maringá (HUM from January 2004 to December 2009. The following Enterobacteriaceae were selected: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter spp. and Proteus mirabilis. All tests were performed according to the recommendations of the Clinical and Laboratory Standards Institute (biochemical identification; susceptibility tests; initial screening and phenotypic confirmatory tests for ESBL. For Enterobacter spp. isolates, a disk approximation test was carried out, adding a cefepime disk. Seven hundred samples were analyzed, and E. coli was the most prevalent bacteria (n= 356. ESBLs were detected phenotypically in 7.3% of E. coli, 61.7% of K. pneumoniae, 33.3% of K. oxytoca, 7.1% of P. mirabilis, and 13.4% of Enterobacter spp samples. Overall ESBL prevalence reached 22% when all producers were taken together. Although HUM is considered a small-sized hospital, it showed high levels of resistance to antimicrobial agents, similar to those observed in bigger hospitals, which demonstrated the need for careful epidemiological surveillance.A produção de beta-lactamases de espectro ampliado (ESBL é considerada um dos mais importantes mecanismos de resistência aos antimicrobianos, o que dificulta o tratamento de infecções causadas por enterobactérias. Dados sobre cultura e testes de sensibilidade foram coletados das fichas do Laboratório de Ensino e Pesquisa de Análises Clínicas de pacientes atendidos no Hospital Universitário de Maringá (HUM, de janeiro de 2004 a dezembro de 2009. As enterobactérias escolhidas foram: Escherichia coli, Klebsiella

  20. PER, CTX-M, TEM and SHV Beta-lactamases in Clinical Isolates of Klebsiella pneumoniae Isolated from Tehran, Iran

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    Leila Nasehi

    2010-06-01

    Full Text Available Objective(sDifferent types of extended spectrum beta-lactamases (ESBLs are encountered in the clinical settings worldwide. There are a few studies regarding the prevalence of ESBL genes among Klebsiella pneumoniae isolates at Tehran especially those of blaPER and blaCTX. The aim of this study was to determine the prevalence of blaSHV, blaTEM ,blaPER and blaCTX genes among clinical K. pneumoniae of different hospitals in Tehran.Materials and MethodsTwo hundred isolates of K. pneumoniae were received from different clinical specimens. The susceptibility of the isolates to 10 different antibiotics was examined by disk diffusion test. The MICs for ceftazidime were also determined using micro-broth dilution assay. Isolates showing MIC 4 μg/ml for ceftazidime were screened for ESBL production by phenotypic confirmatory test (PCT and subjected to PCR for studied genes. Variation among four amplified genes was evaluated using PCR-RFLP.ResultsBy disk diffusion test, resistance to ceftazidime and cefotaxime were 34.7% and 33.5% respectively. However, all strains were susceptible to imipenem. Eighty isolates showed MICs≥ 4 μg/ml for ceftazidime of which 77 (96% were positive for ESBL in PCT. The prevalence of blaSHV, blaCTX-M, blaTEM and blaPER among these isolates were 26%, 24.5%, 18% and 7.5%, respectively. No variation was detected in the genes by PCR-RFLP.ConclusionAs far as we know this is the first report of the blaPER-1 in K. pneumoniae in Iran. The blaCTX-M was the second most common gene detected among the ESBL positive isolates of K. pneumoniae. For rapid identification of ESBL producing isolates it was recommended that clinical laboratories adopt simple test based on CLSI recommendation for confirming ESBL production in enterobacterial species.

  1. Emergence of Escherichia coli Producing Extended-Spectrum AmpC beta-lactamases (ESAC in animals

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    Marisa eHaenni

    2014-02-01

    Full Text Available In both humans and animals, the spread of Extended-Spectrum -Lactamases (ESBL/AmpC producers has become a major issue, particularly due to the plasmidic dissemination of most of these genes. Besides, over-expression of the chromosomal ampC gene was largely reported in human and animal Enterobacteriaceae and, more recently, modifications within the coding region of the ampC gene (encoding Extended-spectrum AmpC -lactamases (ESACs were shown to be responsible for an hydrolysis spectrum expanded to oxyiminocephalosporins in humans. In this study, among 6765 cattle E. coli isolates, 28 (0.37% isolates harboring a reduced susceptibility to cefepime (MICs ranging from 0.5 to 12 g/ml were investigated as presumptive ESACs producers. Highly conserved mutations in the promoter/attenuator region were identified at positions -88, -82, -42, -18, -1 and +58. Using sequencing and cloning experiments, amino acid substitutions of the AmpC beta-lactamase were characterized at positions 287 (mostly S287N, but also S287C, 292 (A292V and 296 (H296P, similarly to data reported in humans. Interestingly, those cattle ESAC-producing E. coli isolates predominantly belonged to the Clonal Complex (CC23, thus mirroring what has been described in humans. The driving forces for the selection of ESACs in animals are unknown, and their prevalence needs to be further investigated in the different animal sectors. Considering the over-representation of ESAC-producing E. coli belonging to CC23 in both humans and animals, exchanges of ESAC producers between the two populations may have occurred as well. To our best knowledge, this study is the first report of ESACs in animals worldwide, which should be considered an emerging mechanism contributing to the resistance to extended-spectrum cephalosporins in the animal population.

  2. Molecular characterization of the extended-spectrum beta-lactamase (ESBL)-producing Shigella spp. in Shanghai.

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    Li, J; Li, B; Ni, Y; Sun, J

    2015-03-01

    Shigellosis is a public health concern in China. We tested 216 Shigella isolates collected in Shanghai in 2007 for the production of extended-spectrum beta-lactamases (ESBLs). ESBL-producing isolates were characterized using polymerase chain reaction (PCR)-based genotyping, conjugation, pulsed-field gel electrophoresis (PFGE), and DNA sequence analysis of regions adjacent to bla genes. Plasmids containing genes encoding ESBLs were analyzed using plasmid replicon typing. ESBLs were produced by 18.1 % (39/216) of Shigella isolates, and all 39 ESBL-producing strains harbored bla CTX-M genes. CTX-M-14 was the most frequent variant (69.2 %, 27/39), followed by CTX-M-15 (15.4 %, 6/39). All bla CTX-M genes were transferable by conjugation, and the insertion sequence ISEcp1 was detected upstream of all bla CTX-M genes. The CTX-M-producing Shigella isolates showed high clonal diversity. IncI1, IncFII, IncN, and IncB/O replicons were respectively detected in 23 (58.9 %), 9 (23.1 %), 1 (2.6 %), and 1 (2.6 %) of the 39 transconjugants carrying bla CTX-M. The bla CTX-M-14 genes were most frequently carried by IncI1 (n = 13, 48.1 %) or IncFII (n = 9, 33.3 %) plasmids, and the bla CTX-M-15 genes were closely associated with IncI1 (n = 5, 83.3 %). Our findings demonstrate the high prevalence of ESBL-producing Shigella in Shanghai, the importance of plasmids and ISEcp1 as carriers of bla CTX-M genes, and the close association between certain bla CTX-M genes with a specific plasmid. PMID:25252628

  3. Prevalence of Extended Spectrum Beta-Lactamases among Escherichia coli and Klebsiella spp isolates in Manipal Teaching Hospital, Pokhara, Nepal

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    Shristi Raut

    2015-06-01

    Full Text Available Objective: To isolate, identify and phenotypically characterize extended spectrum beta-lactamase (ESBL-producing Escherichia coli and Klebsiella spp in Manipal Teaching Hospital. Methods: Cross sectional study was conducted among E. coli and Klebsiella spp recovered from patients’ various samples to establish the prevalence of organisms producing ESBL in Manipal Teaching Hospital, Pokhara, Nepal between October 2011 and April 2012. ESBL production was detected by Clinical laboratory Standards Institute (CLSI recommendations. Results: E. coli (n=285 and Klebsiella spp (n=55 were isolated from various clinical samples. The specimens were urine 255 (75%, blood 18 (5.3%, pus 48 (14.1%, sputum 17 (5.1%, and body fluid 2 (0.6%. Seventy six (22.4% were ESBL producing organisms by phenotypic confirmatory test with double disk diffusion method. ESBL group of organisms showed 100% resistance to ampicillin and cefotaxime. All the organisms in this study were 100% sensitive to imipenem and 95.6% sensitive to cefoperazone+sulbactam combination. ESBL producing isolates showed high rate of resistance to ciprofloxacin (90.7%, ceftriaxone (89.4%, ceftazidime (89.4%, cotrimoxazole (90.4% and norfloxacin (88.1% as compared to non-ESBL group. Conclusion: ESBL producing E. coli and Klebsiella spp showed high prevalence in Nepal. Routine laboratory testing for ESBL in Nepalese hospitals is needed in order to optimize antibiotic management and reduce the risk of spread of infections caused by ESBL producers. J Microbiol Infect Dis 2015;5(2: 69-75

  4. Identification of Klebsiella pneumoniae strains harboring inactive extended-spectrum beta-lactamase antibiotic-resistance genes

    Institute of Scientific and Technical Information of China (English)

    Xu Li; Zhai Yao; Lyu Yuan; Wang Qi; An Shuchang; Chen Jichao; Chen Yusheng

    2014-01-01

    Background The extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae has increasingly become a major contributor to nosocomial infections and can exhibit multiple antibiotic resistance.Previous studies have focused on the resistance genes in ESBL-producing strains,and the resistance-associated genetic environment of non-ESBL-producing strains has been ignored until now.Here,we investigated the occurrence and characteristics of non-ESBL-producing K.pneumoniae,which potentially carries unexpressed resistance genes.Methods K.pneumoniae strains were collected from five medical institutions in China from February 2010 to August 2013.The VITEK-2 ESBL detection system was used as a primary screen to identify the ESBL-producing phenotype,and the three primary types of ESBL-associated genes (CTX,SHV,and TEM) were detected by polymerase chain reaction (PCR) to confirm the strains presenting with a non-ESBL-producing phenotype.mRNA expression in the non-ESBL-producing strains was further screened by reverse-transcription PCR (RT-PCR) to validate their transcriptional efficiency.Results Out of 224 clinically isolated antibiotic-sensitive K.pneumoniae strains with a non-ESBL-producing phenotype,5 (2.2%) were identified to carry inactivated ESBL blaSHV genes with intact upstream promoter regions and resistance gene sequences.Interestingly,three of the five antibiotic-sensitive K.pneumoniae strains containing ESBL blaSHV genes still exhibited mRNA transcription of blasHv,while the other two exhibited no mRNA transcription.Conclusion These findings suggest that inactivated ESBL genes exist in non-ESBL-producing antibiotic-sensitive K.pneumoniae strains,which have the potential to transform the strain into an ESBL phenotype if an inappropriate application or overdose of antibiotics is implemented during clinical management.

  5. Microbial resistance and frequency of extended-spectrum beta-lactamase (ESBL in isolated from blood cultures

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    Ruan Carlos Gomes da Silva

    2014-12-01

    Full Text Available Introduction:The emergence and spread of isolated carriers of extended-spectrum beta-lactamase (ESBL have complicated the treatment of nosocomial infections, since its production is not easily identified by the sensitivity tests, routinely performed in clinical laboratories, leading to difficulties in the hospital control of resistant microorganisms and antibiotics misuse.Objective:The objective of this study was to analyze the resistance profile and the frequency of ESBL in Gram-negative bacteria isolated from blood cultures. A hundred bacterial samples from blood cultures of adult patients were analyzed, which were phenotypically identified by biochemical tests of carbohydrates fermentation and submitted to determination of the resistance profile by disc diffusion test and ESBL screening by disc approximation and disc replacement methods.Results:Among the bacterial samples tested, 30 were identified as Gram-negative bacteria, predominantly by Proteus mirabilis, Pantoea agglomerans, and Escherichia coli. Of these, 73.33% were positive for the detection of ESBL by phenotypic tests, and was found mainly in Pantoea agglomerans, Proteus mirabilis, and Enterobacter cloacae.Conclusion:The increase in the occurrence of ESBL in different Enterobacteriaceae shows the importance of the amplification of detection in other species than Escherichia coli or Klebsiella sp., so that the assistance to the patient is not restrained, since these resistant bacteria cannot be detected by the laboratories. Considering the frequency of ESBL in this study, we highlight the importance of its detection, aiming to its contribution to the development of improvements in the health care policies of hospitals.

  6. High prevalence of extensively drug-resistant and metallo beta-lactamase-producing clinical Acinetobacter baumannii in Iran.

    Science.gov (United States)

    Maspi, Hossein; Mahmoodzadeh Hosseini, Hamideh; Amin, Mohsen; Imani Fooladi, Abbas Ali

    2016-09-01

    Acinetobacter species particularly Acinetobacter baumannii (A. baumannii) have been widely reported as broad-spectrum antibiotic resistant pathogens. Expression of various types of metallo beta-lactamases (MBL), classified as Ambler class B, has been associated with carbapenem resistance. Here, we attempted to assess the frequency of extensively drug-resistant (XDR) and MBL-producing A. baumannii among clinical isolates. 86 clinical A. baumannii strains were collected from 2014 to 2015 and their susceptibility to meropenem (10 μg), imipenem (10 μg), azteronem (30 μg), pipracillin (100 μg) tazobactam (110 μg), tobramycin (10 μg), fosfomycin (200 μg), rifampicin (5 μg), colistin (10 μg), tigecycline (15 μg), sulbactam/ampicillin (10 μg + 10 μg) and polymixin B (300 U) was evaluated using disk diffusion method. The MBL-producing isolates were screened using combined disc diffusion method. Furthermore, the presence of blaVIM, blaIMP, blaSPM, blaGIM, blaSIM and blaNDM was detected by PCR. 34.9% of isolates were recovered from bronchoalveolar lavage (BAL). 81 (94.2%) and 62 (71.2%) isolates were multidrug resistance (MDR) and XDR, respectively. 44 (51.2%) and 65 (75.6%) isolates were MBL-producing strains with resistance to imipenem and meropenem, respectively. 2 (2.3%), 13 (15.1%), 2 (2.3%), 4 (4.7%) and 2 (2.3%) isolates carried blaVIM, blaIMP, blaSPM, blaGIM and blaSIM genes, respectively. Our data showed that the rate of XDR and MBL A. baumannii is on the rise. PMID:27448835

  7. Determination of Extended-Spectrum Beta-lactamases Genes and Antibiotic Resistance Patterns in Escherichia coli Isolates from Healthy Cats

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    Baharak Akhtardanesh

    2016-01-01

    Full Text Available ne"> Background: This study was set to detect extended-spectrum beta-lactamases (ESBLsproducing E. coli isolates and the genes underlying their resistance in relation to phylogeneticbackground from fecal samples of healthy owned cats.Methods: A total of 50 E. coli isolates were confirmed by standard bacteriological tests. Thephylogenetic analyses of the isolates were carried out by combinations of three genetic markerschuA, yjaA and DNA fragment TspE4.C2 by a triplex PCR method. The ESBL (blaCTXM, blaTEM,blaSHV, blaOXA encoding genes were detected. To identify ESBL producing phenotypes, allselected isolates were screened with a double disk synergy test including cefotaxime, cefotaximewith clavulanic acid, ceftazidime and ceftazidime with clavulanic acid.Results: Results showed that E. coli isolates fell into four phylogenetic groups (A, D, B1 andB2 with prevalence of 78%, 4%, 8%, 10% and five phylogenetic subgroups including A0 (74%, A1 (4 %, B1 (8 %, B2–2 (6 %, B2–3 (4 % and D1 (4 %, respectively. Among all E. coliisolates, 4% were positive for bla SHV, blaCTX-M-15 and blaOXA-1 genes which distributed in B2-2,B2-3, A0 subgroups, respectively. According to antibiotic susceptibility test, 20 isolates wereresistant which belonged to D (D1 phylogenetic subgroup and A (A0 phylogenetic subgroupgroups.Conclusion: The results showed that healthy cats could be considered as potential source for thedissemination of ESBL-encoding genes. Further investigations in companion animals and theirowners are needed to clarify the importance of spreading of these zoonotic strains.

  8. Class 1 integrons contributes to antibiotic resistance among clinical isolates of Escherichia coli producing extended-spectrum beta-lactamases

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    T Chen

    2013-01-01

    Full Text Available Objectives: The objective of this study is to determine the prevalence of antibiotic resistance factors, including the production of extended-spectrum beta-lactamases (ESBLs and the presence of class 1 integrons among Escherichia coli isolated from clinical specimens. Materials and Methods: Bacterial species identification was performed using a VITEK-2 system (VITEK2 GN-card; bioMérieux, France. Antimicrobial susceptibility testing was determined using the disk diffusion method according to the 2010 Clinical and Laboratory Standards Institute guidelines. Polymerase chain reaction (PCR was used to detect integrons and amplify variable regions of the bla TEM, bla SHV and bla CTX-M genes. Gene cassettes were detected by deoxyribonucleic acid sequencing. Results: In this study, 58% (100/172 of clinical E. coli isolates were identified as ESBL producers. We found that 90% of the ESBL-producing E. coli isolates harbored the blaCTX-M gene, whereas only 59% and 32% possessed the blaTEM and blaSHV genes respectively. The presence of class 1 integrons was based on the detection of the integrase gene by PCR. A total of 69% of the ESBL-producing isolates were integron-positive. Resistance to 10 antibiotics, including quinolones, sulfonamides and β-lactam/enzyme inhibitors, was significantly higher in the class 1 integron-positive isolates (P < 0.05. The occurrence of class 1 integrons in blaTEM , blaSHV and blaCTX-M gene carriers was 72.9%, 84.4% and 68.9%, respectively. Class 1 integrons were detected in 61.5% of the isolates with only one ESBL genotype, but in 69.0% and 92.3% of the isolates with two or three different ESBL genotypes, respectively. Conclusions: Our findings indicate that clinical strains of bacteria with multiple ESBL genotypes may have greater opportunities to carry class 1 integrons.

  9. Extended-spectrum beta-lactamases producing E. coli in wildlife, yet another form of environmental pollution?

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    Sebastian eGuenther

    2011-12-01

    Full Text Available Wildlife is normally not exposed to antimicrobial agents but can acquire antimicrobial resistant bacteria through contact with humans, domesticated animals and the environment, where water polluted with faeces seems to be the most important vector. E. coli, a ubiquitous commensal bacterial species colonizing the intestinal tract of mammals and birds, is also found in the environment. Extended-spectrum beta-lactamases producing E. coli (ESBL-E. coli represent a major problem in human and veterinary medicine, particular in nosocomial infections. Additionally an onset of community acquired ESBL-E. coli infections and an emergence in livestock farming has been observed in recent years, suggesting a successful transmission as well as persistence of ESBL-E. coli strains outside clinical settings. Another parallel worldwide phenomenon is the spread of ESBL-E. coli into the environment beyond human and domesticated animal populations, and this seems to be directly influenced by antibiotic practice. This might be a collateral consequence of the community onset of ESBL-E. coli infections but can result (a in a subsequent colonization of wild animal populations which can turn into an infectious source or even a reservoir of ESBL-E.coli, (b in a contribution of wildlife to the spread and transmission of ESBL-E. coli into fragile environmental niches, (c in new putative infection cycles between wildlife, domesticated animals and humans, and (d in problems in the medical treatment of wildlife. This review aims to summarize the current knowledge on ESBL-E. coli in wildlife, in turn underlining the need for more large scale investigations, in particular sentinel studies to monitor the impact of multiresistant bacteria on wildlife.

  10. Neutron Diffraction Studies of a Class A beta-Lactamase Toho-1 E166A/R274N/R276N Triple Mutant

    Energy Technology Data Exchange (ETDEWEB)

    Blakeley, Matthew P. [Institut Laue-Langevin (ILL); Chen, Yu [ORNL; Afonine, Pavel [Lawrence Berkeley National Laboratory (LBNL)

    2010-01-01

    beta-Lactam antibiotics have been used effectively over several decades against many types of bacterial infectious diseases. However, the most common cause of resistance to the beta-lactam antibiotics is the production of beta-lactamase enzymes that inactivate beta-lactams by rapidly hydrolyzing the amide group of the beta-lactam ring. Specifically, the class A extended-spectrum beta-lactamases (ESBLs) and inhibitor-resistant enzymes arose that were capable of hydrolyzing penicillins and the expanded-spectrum cephalosporins and monobactams in resistant bacteria, which lead to treatment problems in many clinical settings. A more complete understanding of the mechanism of catalysis of these ESBL enzymes will impact current antibiotic drug discovery efforts. Here, we describe the neutron structure of the class A, CTX-M-type ESBL Toho-1 E166A/R274N/R276N triple mutant in its apo form, which is the first reported neutron structure of a beta-lactamase enzyme. This neutron structure clearly reveals the active-site protonation states and hydrogen-bonding network of the apo Toho-1 ESBL prior to substrate binding and subsequent acylation. The protonation states of the active-site residues Ser70, Lys73, Ser130, and Lys234 in this neutron structure are consistent with the prediction of a proton transfer pathway from Lys73 to Ser130 that is likely dependent on the conformation of Lys73, which has been hypothesized to be coupled to the protonation state of Glu166 during the acylation reaction. Thus, this neutron structure is in agreement with a proposed mechanism for acylation that identifies Glu166 as the general base for catalysis.

  11. Antibacterial and toxicological evaluation of beta-lactams synthesized by immobilized beta-lactamase-free penicillin amidase produced by Alcaligenes sp.

    Science.gov (United States)

    Gayen, Jiaur R; Majee, Sutapa B; Das, Shuvendu; Samanta, Timir B

    2007-12-01

    Search for anti-beta-lactamase and synthesis of newer penicillin were suggested to overcome resistance to penicillin in chemotherapy. It was found that clavulanic acid, an ant-beta-lactamase was ineffective due to its structural modification by bacteria. Thus, there is a need for the synthesis of newer pencillins. Retro-synthesis was inspired by the success of forward reaction i.e.conversion of penicillin G to 6-aminopenicillanic acid (6-APA) by biological process. In the present study a better enzymatic method of synthesis of newer pencillin by a beta-lactamase-free penicillin amidase produced by Alcaligenes sp. is attempted. Antibacterial and toxicological evaluation of the enzymatically synthesized beta-lactams are reported. Condensation of 6-APA with acyl donor was found to be effective when the reaction is run in dimethyl formamide (DMF 50% v/v) in acetate buffer (25 mM pH 5.0) at 37 degrees C. Periplasm entrapped in calcium alginate exihibited the highest yield (approximately 34%) in synthesis. The minimum inhibitory concentration of the synthetic products against Staphylococcus aureus and Salmonella typhi varied between 20-80 microg/ml. Some of the products exhibited antibacterial activity against enteric pathogens. It was interesting to note that product A was potent like penicillin G. LD50 value of three products (product A, B and C) was more than 12 mg/kg. Furthermore, these synthetic beta-lactams did not exihibit any adverse effect on house keeping enzymes viz., serum glutamate oxalacetate-trans-aminase, serum glutamate pyruvate -trans-aminase, acid phosphatase, alkaline phosphatase of the test animals. The hematological profile (RBC and WBC) of the test animals also remained unaffected. PMID:18254214

  12. OXA-46, a new class D beta-lactamase of narrow substrate specificity encoded by a blaVIM-1-containing integron from a Pseudomonas aeruginosa clinical isolate.

    Science.gov (United States)

    Giuliani, Francesco; Docquier, Jean-Denis; Riccio, Maria Letizia; Pagani, Laura; Rossolini, Gian Maria

    2005-05-01

    A novel OXA-type enzyme, named OXA-46, was found to be encoded by a gene cassette inserted into a class 1 integron from a multidrug-resistant Pseudomonas aeruginosa clinical isolate. The variable region of the integron also contained a bla(VIM-1) metallo-beta-lactamase cassette and a duplicated aacA4 aminoglycoside acetyltransferase cassette. OXA-46 belongs to the OXA-2 lineage of class D beta-lactamases. It exhibits 78% sequence identity with OXA-2 and the highest similarity (around 92% identity) with another OXA-type enzyme detected in clinical isolates of Burkholderia cepacia and in unidentified bacteria from a wastewater plant. Expression of bla(OXA-46) in Escherichia coli decreased susceptibility to penicillins and narrow-spectrum cephalosporins but not to extended-spectrum cephalosporins, cefsulodin, aztreonam, or carbapenems. The enzyme was overproduced in E. coli and purified by two anion-exchange chromatography steps (approximate yield, 6 mg/liter). OXA-46 was made of a 28.5-kDa polypeptide and exhibited an alkaline pI (7.8). In its native form OXA-46 appeared to be dimeric, and the oligomerization state was not affected by EDTA. Kinetic analysis of OXA-46 revealed a specificity for narrow-spectrum substrates, including oxacillin, other penicillins (but not temocillin), and narrow-spectrum cephalosporins. The enzyme apparently did not interact with temocillin, oxyimino-cephalosporins, or aztreonam. OXA-46 was inactivated by tazobactam and carbapenems and, although less efficiently, also by clavulanic acid. Enzyme activity was not affected either by EDTA or by divalent cations and exhibited low susceptibility to NaCl. These findings underscore the functional and structural diversity that can be encountered among class D beta-lactamases. PMID:15855521

  13. Molecular characterization and clinical significance of New Delhi metallo-beta-lactamases-1 producing Escherichia coli recovered from a South Indian tertiary care hospital

    Directory of Open Access Journals (Sweden)

    Arindam Ckakraborty

    2015-01-01

    Full Text Available Context: The increased rate of infection by New Delhi metallo-beta-lactamases-1 (NDM1 producing Escherichia coli is a major concern since they show a high rate of drug resistance and are responsible for mortality and morbidity. Aims: To characterize the NDM1 producing E. coli isolates and their impact on patients′ clinical outcome. Settings and Design: This descriptive study was carried out in a multi-specialty tertiary care hospital. Materials and Methods: Three hundred nonrepeat strains of E. coli from inpatients were included in the study. Modified Hodge test and metallo-beta-lactamases (MBL e-test were performed to detect carbapenemase and MBL activity. Polymerase chain reaction (PCR technique was performed to detect NDM1. NDM1 positive isolates were further tested for plasmid mediated AmpC, blaCTX , blaSHV , blaTEM genes and also for phylogrouping by PCR methods. Treatment and patients′ clinical outcome were also analyzed. Results: Out of 300 isolates, 21 (7% were MBL producers by phenotypic methods. Of this, 17 (81% were NDM1 positives, among the NDM1 producers 6 (35% isolates were belongs to phylogroups D followed by A 5 (29%, B1 4 (24% and B2 2 (12%, 15 (88% isolates were blaCTX-M positive suggestive of extended-spectrum beta lactamase producing strain and 7 (47% were positive with CIT type of AmpC. With the follow-up of the patients, it was found that 12 (71% recovered and 3 (18% developed relapses, and mortality was seen in 2 (12% patients. Conclusions: NDM1 producing isolates showed a high degree of drug resistance but can be treated with suitable antimicrobials, in the majority. Early detection and choice of appropriate antibiotics may help in reducing mortality and morbidity.

  14. Detection of extended spectrum beta-lactamases in gram negative bacilli from clinical specimens in a teaching hospital in South eastern Nigeria

    Directory of Open Access Journals (Sweden)

    C N Akujobi

    2010-01-01

    Full Text Available Antimicrobial drug resistance seen among many gram-negative bacteria, especially those expressing the extended-spectrum β- lactamase (ESBL enzymes that hydrolyze the expanded- spectrum cephalosporins has been on the increase. This has compromised treatment options and thus a threat to the containment of bacterial infections. To determine the existence of the extended-spectrum β-lactamase enzymes in Nnewi, 250 clinical isolates of members of the family Enterobacteriaceae and Pseudomonas species from Nnamdi Azikiwe University Teaching Hospital, Nnewi were identified by conventional methods. These include Klebsiella species (96, E. coli (90, Pseudomonas species (37, Enterobacter species (13, Proteus species (6, Citrobacter species (5 and Salmonella species (3. Antimicrobial drug susceptibility testing was carried out on all the isolates by the disc diffusion method. Extended Spectrum Beta- lactamases were detected by the double disc synergy test. High level of antimicrobial resistance was noted in test organisms against some of the antimicrobial drugs: Ampicillin + Cloxacillin (93.2%, Tetracycline (90.8%, Streptomycin (82.4%, and Nalidixic acid (62%, and low level of resistance was observed against Ofloxacin (26.4%, Cefotaxime (28.8% and Nitrofurantoin (28.8%. One hundred and forty four isolates (57.6% were suspected ESBL-producers judged by their resistance to any of the third generation cephalosporins used but 40 (16% actually produced the extended spectrum beta- lactamase enzymes. This shows the existence of Extended Spectrum Beta- Lactamase producing gram negative organisms in Nnewi. Considering the treatment difficulties, as well as the high cost of treatment associated with these organisms, concerted efforts are needed to contain their spread.

  15. Structure of GES-1 at Atomic Resolution: Insights Into the Evolution of Carbapenamase Activity in the Class a Extended-Spectrum Beta-Lactamases

    Energy Technology Data Exchange (ETDEWEB)

    Smith, C.A.; /SLAC, SSRL; Caccamo, M.; /Notre Dame U.; Kantardjieff, K.A.; /Cal State, Fullerton; Vakulenko, S.; /Notre Dame U.

    2007-10-08

    The structure of the class A extended-spectrum {beta}-lactamase GES-1 from Klebsiella pneumoniae has been determined to 1.1 Angstrom resolution. GES-1 has the characteristic active-site disulfide bond of the carbapenemase family of {beta}-lactamases and has a structure that is very similar to those of other known carbapenemases, including NMC-A, SME-1 and KPC-2. Most residues implicated in the catalytic mechanism of this class of enzyme are present in the GES-1 active site, including Ser70, which forms a covalent bond with the carbonyl C atom of the {beta}-lactam ring of the substrate during the formation of an acyl-enzyme intermediate, Glu166, which is implicated as both the acylation and deacylation base, and Lys73, which is also implicated as the acylation base. A water molecule crucial to catalysis is observed in an identical location as in other class A {beta}-lactamases, interacting with the side chains of Ser70 and Glu166. One important residue, Asn170, also normally a ligand for the hydrolytic water, is missing from the GES-1 active site. This residue is a glycine in GES-1 and the enzyme is unable to hydrolyze imipenem. This points to this residue as being critically important in the hydrolysis of this class of {beta}-lactam substrate. This is further supported by flexible-docking studies of imipenem with in silico-generated Gly170Asn and Gly170Ser mutant GES-1 enzymes designed to mimic the active sites of imipenem-hydrolyzing point mutants GES-2 and GES-5.

  16. Neutron Diffraction Studies of a Class A beta-Lactamase Toho-1 E166A/R274N/R276N Triple Mutant

    International Nuclear Information System (INIS)

    beta-Lactam antibiotics have been used effectively over several decades against many types of bacterial infectious diseases. However, the most common cause of resistance to the beta-lactam antibiotics is the production of beta-lactamase enzymes that inactivate beta-lactams by rapidly hydrolyzing the amide group of the beta-lactam ring. Specifically, the class A extended-spectrum beta-lactamases (ESBLs) and inhibitor-resistant enzymes arose that were capable of hydrolyzing penicillins and the expanded-spectrum cephalosporins and monobactams in resistant bacteria, which lead to treatment problems in many clinical settings. A more complete understanding of the mechanism of catalysis of these ESBL enzymes will impact current antibiotic drug discovery efforts. Here, we describe the neutron structure of the class A, CTX-M-type ESBL Toho-1 E166A/R274N/R276N triple mutant in its apo form, which is the first reported neutron structure of a beta-lactamase enzyme. This neutron structure clearly reveals the active-site protonation states and hydrogen-bonding network of the apo Toho-1 ESBL prior to substrate binding and subsequent acylation. The protonation states of the active-site residues Ser70, Lys73, Ser130, and Lys234 in this neutron structure are consistent with the prediction of a proton transfer pathway from Lys73 to Ser130 that is likely dependent on the conformation of Lys73, which has been hypothesized to be coupled to the protonation state of Glu166 during the acylation reaction. Thus, this neutron structure is in agreement with a proposed mechanism for acylation that identifies Glu166 as the general base for catalysis.

  17. The Survey of Genes Encoding Beta-Lactamases, in Escherichia Coli Resistant to Beta-Lactam and Non-Beta-Lactam Antibiotics

    Directory of Open Access Journals (Sweden)

    Fereshteh Shahcheraghi

    2010-01-01

    Full Text Available Resistance to the new generation of cephalosporins which is mediated by Extended-Spectrum beta-lactamases (ESBLs has been found amongEscherichia coli isolates throughout the world. These resistance genes and their producers, the micro-organisms carrying beta-lactamases, are responsible for serious clinical and therapeutic problems among inpatients and it is necessary to pay more attention to detection of ESBLs producing organisms.Materiasl and MethodsCollectively 260 isolates of E. coli were obtained from 6 hospitals in Tehran (Iran during April-2006 to April-2007. The antibiotic susceptibility patterns of isolates were determined by disk diffusion method. phenotypic confirmatory test (PCT was carried out for screening of ESBLs. Microbroth dilution assay was used to determine the minimum inhibitory concentration (MIC of ceftazidime. Isolates showing MIC≥2 μg/ml were subjected to polymerase chain reaction (PCR targeting blaTEM, blaSHV, blaCTX and blaPER genes. ResultsThe PCT showed that 48.08% of isolates are ESBL producers (125 of 260. The majority of cefotaxime resistant (90.8% and ceftazidime resistant (92.5% isolates were ESBL producers. The obtained results by PCR revealed that 5.77% (n=15 of 260 and 24.23 (n=63 of isolates can produce SHV and TEM type enzymes respectively. blaCTX was detected in 20.38% of isolates (n=53 and none of them could produce blaPER type beta-lactamases. ConclusionThe results of our study showed that the ESBL genes have high prevalence among clinical isolates of E. coli. Such high dissemination of ESBLs is a serious problem for public health and therefore, it's necessary to seek a program for monitoring ESBLs in hospitals.

  18. Molecular characterization of vancomycin-resistant enterococci and extended-spectrum beta-lactamase-containing Escherichia coli isolates in wild birds from the Azores Archipelago

    OpenAIRE

    Silva, Nuno; Igrejas, Gilberto; Rodrigues, Pedro; Rodrigues, Tiago; Gonçalves, Alexandre; Felgar, Ana; Pacheco, Rui; Gonçalves, David; Cunha, Regina; Poeta, Patrícia

    2011-01-01

    To study the prevalence of vancomycin-resistant enterococci (VRE) and extended-spectrum beta-lactamases (ESBLs) containing Escherichia coli isolates, and the mechanisms of resistance implicated, 220 faecal samples from wild birds were collected between 2006 and 2010 in the Azores Archipelago. Samples were spread in Slanetz-Bartley agar plates supplemented with 4 mg/L of vancomycin and in Levine agar plates supplemented with 2 mg/L of cefotaxime for VRE and ESBL-containing E. coli isolation, r...

  19. Prevalence and Characteristics of the Epidemic Multiresistant Escherichia coli ST131 Clonal Group among Extended-Spectrum Beta-Lactamase-Producing E. coli Isolates in Copenhagen, Denmark

    OpenAIRE

    Olesen, Bente; Hansen, Dennis S.; Nilsson, Frida; Frimodt-Møller, Jakob; Leihof, Rikke Fleron; Struve, Carsten; Scheutz, Flemming; Johnston, Brian; Krogfelt, Karen A.; Johnson, James R.

    2013-01-01

    We report the characteristics of 115 extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli clinical isolates, from 115 unique Danish patients, over a 1-year study interval (1 October 2008 to 30 September 2009). Forty-four (38%) of the ESBL isolates represented sequence type 131 (ST13)1, from phylogenetic group B2. The remaining 71 isolates were from phylogenetic groups D (27%), A (22%), B1 (10%), and B2 (3%). Serogroup O25 ST131 isolates (n = 42; 95% of ST131) comprised 7 differe...

  20. Characterization of paired mucoid/non-mucoid Pseudomonas aeruginosa isolates from Danish cystic fibrosis patients: antibiotic resistance, beta-lactamase activity and RiboPrinting

    DEFF Research Database (Denmark)

    Ciofu, O; Fussing, V; Bagge, N;

    2001-01-01

    The purpose of this study was to characterize 42 paired mucoid and non-mucoid Danish cystic fibrosis (CF) Pseudomonas aeruginosa isolates collected in 1997, by RiboPrinting, antibiotic susceptibility and beta-lactamase activity. Eight P. aeruginosa isolates collected before 1991 were included for...... before 1991 had an antibiotic susceptibility pattern similar to the 1997 isolates. Despite prolonged and intensive antibiotic treatment, susceptible mucoid isolates were isolated from the CF sputum, possibly because these bacteria are protected from the selective pressure of antibiotics by the resistant...

  1. Antimicrobial resistance testing of verocytotoxin-producing Escherichia coli and first description of TEM-52 extended-spectrum {beta}-lactamase in serogroup O26.

    OpenAIRE

    Buvens, Glenn; Bogaerts, Pierre; Glupczynski, Gerald; Lauwers, Sabine; Piérard, Denis

    2010-01-01

    We have investigated the antimicrobial resistance of verocytotoxin-producing Escherichia coli (VTEC) strains isolated from humans, animals, food and environment in Belgium. Resistance was more frequent in human non-O157 as compared to O157 strains from humans or other sources and within non-O157 VTEC, intimin-positive strains were more resistant than intimin-negative strains. We also report the first VTEC producing an IncI1 plasmid-borne blaTEM-52 extended-spectrum beta-lactamase previously a...

  2. Use of ampicillin-sulbactam for treatment of experimental meningitis caused by a beta-lactamase-producing strain of Escherichia coli K-1.

    OpenAIRE

    Guerra-Romero, L; Kennedy, S L; Fournier, M A; Tureen, J H; Täuber, M G

    1991-01-01

    We evaluated the pharmacokinetics and therapeutic efficacy of ampicillin combined with sulbactam in a rabbit model of meningitis due to a beta-lactamase-producing strain of Escherichia coli K-1. Ceftriaxone was used as a comparison drug. The MIC and MBC were 32 and greater than 64 micrograms/ml (ampicillin), greater than 256 and greater than 256 micrograms/ml (sulbactam), 2.0 and 4.0 micrograms/ml (ampicillin-sulbactam [2:1 ratio, ampicillin concentration]) and 0.125 and 0.25 micrograms/ml (c...

  3. Beta-lactam induction of ISEcp1B-mediated mobilization of the naturally occurring bla(CTX-M) beta-lactamase gene of Kluyvera ascorbata.

    OpenAIRE

    Nordmann, Patrice; Lartigue, Marie-Frédérique; Poirel, Laurent

    2008-01-01

    ISEcp1B is an insertion element associated with the emerging expanded-spectrum beta-lactamase bla(CTX-M) genes in Enterobacteriaceae. Because ISEcp1B-bla(CTX-M)positive strains may be identified from humans and animals, the ability of this insertion sequence to mobilize the bla(CTX-M-2) gene was tested from its progenitor Kluyvera ascorbata to study the effects of amoxicillin/clavulanic and cefquinome as enhancers of transposition. These beta-lactam molecules are administered parenterally to ...

  4. Ceftriaxone-sulbactam combination in rabbit endocarditis caused by a strain of Klebsiella pneumoniae producing extended-broad-spectrum TEM-3 beta-lactamase.

    OpenAIRE

    Caron, F.; Gutmann, L; Bure, A; B. Pangon; Vallois, J M; Pechinot, A; Carbon, C

    1990-01-01

    We studied the activity of the combination of sulbactam and ceftriaxone against a Klebsiella pneumoniae strain producing TEM-3, a new extended-broad-spectrum beta-lactamase, in an endocarditis model. In vitro, ceftriaxone was strongly inactivated in the presence of TEM-3 (MBC, 128 micrograms/ml with an inoculum of 5 x 10(5) CFU/ml). A marked inoculum effect was demonstrated with sulbactam: effective concentrations of inhibitor needed to reduce the MIC and MBC of ceftriaxone to similar levels ...

  5. Comparison of the activity of imipenem and beta-lactams combined with sulbactam and clavulanic acid in beta-lactamase-producing strains of Bacteroides fragilis.

    Science.gov (United States)

    Martín, M A; Castillo, A M; Liébana, J; Marín, A; Alados, J C; Piédrola, G

    1991-01-01

    We compared the "in vitro" activity of imipenem with 14 beta-lactams, both alone and in combination with clavulanic acid, and sulbactam against 110 beta-lactamase-producing strains of Bacteroides fragilis. The following antibiotics were tested: amoxycillin, penicillin, mezlocillin, piperacillin, cephalothin, cephazolin, cefamandole, cefmetazole, cefonicid, cefoxitin, cefotaxime, ceftazidime, ceftizoxime, and ceftriaxone. In all cases, except those of cefoxitin and cefmetazole, these combinations showed a statistically significant increase in beta-lactam activity, which was, however, never higher than that of imipenem, the antibiotic which performed best against Bacteroides fragilis. PMID:1940333

  6. Meta-analysis of proportion estimates of Extended-Spectrum-Beta-Lactamase-producing Enterobacteriaceae in East Africa hospitals

    DEFF Research Database (Denmark)

    Sonda, Tolbert; Kumburu, Happiness; van Zwetselaar, Marco;

    2016-01-01

    Background: A high proportion of Extended-Spectrum-Beta-Lactamase (ESBL) producing Enterobacteriaceae is causing common infections in all regions of the world. The burden of antibiotic resistance due to ESBL in East Africa is large but information is scarce and thus it is unclear how big the...... due to ESBL between countries. This highlights a need for active surveillance systems which can help understand the actual epidemiology of ESBL, aid in formulating national or regional guidelines for proper screening of ESBL, and support developing standardized approaches for managing patients...

  7. Phenotypic and molecular characterization of CTX-M extended-spectrum beta-lactamase-producing Escherichia coli isolates in Shiraz, Iran

    OpenAIRE

    Khosrow Zamani; Amir Emami; Abdollah Bazargani; Afagh Moattari

    2015-01-01

    AbstractINTRODUCTIONThe aim of this study was to detect the prevalence of the extended-spectrum beta-lactamase (ESBL)-encoding CTX-M gene in Escherichia coliisolates.METHODS:Phenotypic screening of 376 E. coli isolates for ESBL was conducted using disk diffusion. ESBL-producing isolates were tested using PCR and specific primers. The blaCTX-M cluster was identified using the RFLP method, and its genotype was sequenced.RESULTS:From 202 ESBL-producing E. coli , 185 (91.5%) possessed CTX-M genes...

  8. Impact of Fecal Carriage of Extended-Spectrum Beta-Lactamase Producing Enterobacteriaceae on the Outcomes of Transrectal Needle Biopsies of the Prostate.

    OpenAIRE

    Ergönül, Önder; Tükenmez-Tigen, Elif; Altinkanat, Gülşen; Özgen, Mahir Bülent; Ertük Şengel, Buket; Odabaşi, Zekaver; Türkerí, Levent N.; Mülazimo?lu, Lütfye; Korten, Volkan

    2013-01-01

    Objective: This study aimed to address the prevalence, the risk factors and the results of fecal carriage of extended-spectrum beta-lactamase producing Enterobacteriaceae (ESBL-PE) in patients who had undergone transrectal needle biopsy of the prostate (TRNBP). Patients and Methods: A total of 143 patients who had undergone TRNBP were included. Of these, 33(23%) had fecal carriage of ESBL-PE. The mean age of the patients was 62 ± 7.5 (43-81) years. Results: A univariate analysis showed that q...

  9. Ceftazidime/avibactam: a novel cephalosporin/nonbeta-lactam beta-lactamase inhibitor for the treatment of complicated urinary tract infections and complicated intra-abdominal infections.

    Science.gov (United States)

    Hidalgo, Jose A; Vinluan, Celeste M; Antony, Nishaal

    2016-01-01

    There has been greater interest in developing additional antimicrobial agents due to the increasing health care costs and resistance resulting from bacterial pathogens to currently available treatment options. Gram-negative organisms including Enterobacteriaceae and Pseudomonas aeruginosa are some of the most concerning threats due to their resistance mechanisms: extended-spectrum beta-lactamase production and Klebsiella pneumoniae carbapenemase enzymes. Ceftazidime is a third-generation broad-spectrum cephalosporin with activity against P. aeruginosa and avibactam is a novel nonbeta-lactam beta-lactamase inhibitor. Avycaz(®), the trade name for this new combination antibiotic, restores the activity of ceftazidime against some of the previously resistant pathogens. Avycaz was approved in 2015 for the treatment of complicated urinary tract infections, including pyelonephritis, and complicated intra-abdominal infections with the addition of metronidazole in patients with little to no other treatment options. This review article assesses the clinical trials and data that led to the approval of this antibiotic, in addition to its spectrum of activity and limitations. PMID:27528799

  10. Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-Producing Escherichia coli among Uropathogens of Pediatrics in North of Iran

    Directory of Open Access Journals (Sweden)

    Mohammad Sadegh Rezai

    2015-01-01

    Full Text Available Escherichia coli remains 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-producing E. coli isolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of the E. coli isolates 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 of CTX, TEM, SHV, GES, and VEB beta-lactamase genes. About 30.5% of isolated E. coli was ESBL-producing strain. The TEM gene was the most prevalent (49% followed by SHV (44%, CTX (28%, VEB (8%, and GES (0% genes. The ESBL-producing E. coli isolates 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-producing E. coli in 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.

  11. Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-Producing Escherichia coli among Uropathogens of Pediatrics in North of Iran.

    Science.gov (United States)

    Rezai, Mohammad Sadegh; Salehifar, Ebrahim; Rafiei, Alireza; Langaee, Taimour; Rafati, Mohammadreza; Shafahi, Kheironesa; Eslami, Gohar

    2015-01-01

    Escherichia coli remains 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-producing E. coli isolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of the E. coli isolates 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 of CTX, TEM, SHV, GES, and VEB beta-lactamase genes. About 30.5% of isolated E. coli was ESBL-producing strain. The TEM gene was the most prevalent (49%) followed by SHV (44%), CTX (28%), VEB (8%), and GES (0%) genes. The ESBL-producing E. coli isolates 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-producing E. coli in 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. PMID:26064896

  12. Loop-Mediated Isothermal Amplification untuk Mendeteksi Gen blaTEM sebagai Penyandi Extended-Spectrum Beta-Lactamase pada Isolat Enterobacteriaceae

    Directory of Open Access Journals (Sweden)

    Bayu A. P. Wilopo

    2015-12-01

    Full Text Available Extended-spectrum beta-lactamase (ESBL is a beta-lactamase enzyme that is capable of hydrolyzing penicillin, cephalosporin, and monobactam, and can be inhibited by clavulanic acid. This enzyme is encoded by multiple genes, one of them is blaTEM. Polymerase chain reaction (PCR is one of the DNA amplification methods that are frequently used; however, there are other methods that can be used including, among others, loop-mediated isothermal amplification (LAMP. LAMP requires simple equipment with quicker and easy-to-read results compared to PCR. This study was a diagnostic test to explore the sensitivity and specificity of LAMP method compared to PCR in detecting blaTEM gene. Furthermore, the concordance between LAMP and PCR methods was assessed. A total of 92 Enterobacteriaceae isolates were examined by PCR and LAMP methods and compared. The result showed that the LAMP method had a sensitivity of 91.4% and a specificity of 91.2% with a concordance value (kappa of 85.4%. In conclusion, LAMP method has a good validity and a very good conformity compared to the PCR method. Therefore, LAMP method can be used as an alternative diagnostic test, especially in limited settings.

  13. Ceftazidime/avibactam: a novel cephalosporin/nonbeta-lactam beta-lactamase inhibitor for the treatment of complicated urinary tract infections and complicated intra-abdominal infections

    Science.gov (United States)

    Hidalgo, Jose A; Vinluan, Celeste M; Antony, Nishaal

    2016-01-01

    There has been greater interest in developing additional antimicrobial agents due to the increasing health care costs and resistance resulting from bacterial pathogens to currently available treatment options. Gram-negative organisms including Enterobacteriaceae and Pseudomonas aeruginosa are some of the most concerning threats due to their resistance mechanisms: extended-spectrum beta-lactamase production and Klebsiella pneumoniae carbapenemase enzymes. Ceftazidime is a third-generation broad-spectrum cephalosporin with activity against P. aeruginosa and avibactam is a novel nonbeta-lactam beta-lactamase inhibitor. Avycaz®, the trade name for this new combination antibiotic, restores the activity of ceftazidime against some of the previously resistant pathogens. Avycaz was approved in 2015 for the treatment of complicated urinary tract infections, including pyelonephritis, and complicated intra-abdominal infections with the addition of metronidazole in patients with little to no other treatment options. This review article assesses the clinical trials and data that led to the approval of this antibiotic, in addition to its spectrum of activity and limitations. PMID:27528799

  14. Surveillance of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Dairy Cattle Farms in the Nile Delta, Egypt

    Science.gov (United States)

    Braun, Sascha D.; Ahmed, Marwa F. E.; El-Adawy, Hosny; Hotzel, Helmut; Engelmann, Ines; Weiß, Daniel; Monecke, Stefan; Ehricht, Ralf

    2016-01-01

    Introduction: Industrial livestock farming is a possible source of multi-resistant Gram-negative bacteria, including producers of extended spectrum beta-lactamases (ESBLs) conferring resistance to 3rd generation cephalosporins. Limited information is currently available on the situation of ESBL producers in livestock farming outside of Western Europe. A surveillance study was conducted from January to May in 2014 in four dairy cattle farms in different areas of the Nile delta, Egypt. Materials and Methods: In total, 266 samples were collected from 4 dairy farms including rectal swabs from clinically healthy cattle (n = 210), and environmental samples from the stalls (n = 56). After 24 h pre-enrichment in buffered peptone water, all samples were screened for 3rd generation cephalosporin-resistant Escherichia coli using Brilliance™ ESBL agar. Suspected colonies of putatively ESBL-producing E. coli were sub-cultured and subsequently genotypically and phenotypically characterized. Susceptibility testing using the VITEK-2 system was performed. All suspect isolates were genotypically analyzed using two DNA-microarray based assays: CarbDetect AS-1 and E. coli PanType AS-2 kit (ALERE). These tests allow detection of a multitude of genes and their alleles associated with resistance toward carbapenems, cephalosporins, and other frequently used antibiotics. Serotypes were determined using the E. coli SeroGenotyping AS-1 kit (ALERE). Results: Out of 266 samples tested, 114 (42.8%) ESBL-producing E. coli were geno- and phenotypically identified. 113 of 114 phenotypically 3rd generation cephalosporin-resistant isolates harbored at least one of the ESBL resistance genes covered by the applied assays [blaCTX-M15 (n = 105), blaCTX-M9 (n = 1), blaTEM (n = 90), blaSHV (n = 1)]. Alarmingly, the carbapenemase genes blaOXA-48 (n = 5) and blaOXA-181 (n = 1) were found in isolates that also were phenotypically resistant to imipenem and meropenem. Using the array-based serogenotyping

  15. Commensal Enterobacteriaceae as reservoirs of extended-spectrum beta-lactamases, integrons and sul genes in Portugal

    Directory of Open Access Journals (Sweden)

    Elisabete eMachado

    2013-04-01

    Full Text Available Bacteria colonizing the human intestine have a relevant role in the spread of antimicrobial resistance. We investigated the faecal carriage of extended-spectrum beta-lactamase (ESBL-producing Enterobacteriaceae in healthy humans from Portugal and analysed the distribution of sul genes and class 1 and 2 integrons. Faecal samples (n=113 were recovered from healthy persons (North/Centre of Portugal, 2001-04 and plated on MacConkey agar with and without ceftazidime (1mg/L or cefotaxime (1mg/L. Isolates representing different morphotypes/plate and antibiotic susceptibility patterns (n=201 were selected. Isolates resistant to sulfonamides and/or streptomycin, gentamicin and trimethoprim were screened (PCR, sequencing for sul genes (sul1, sul2, sul3 and class 1 and 2 integrons. Presence of ESBLs was inferred using the DDST and further confirmed by PCR and sequencing. ESBL producers were selected for clonal analysis, plasmid characterization and conjugation assays by standard methods. ESBL-producing isolates were found in 1.8% (2/113 of samples, corresponding to Escherichia coli of phylogroups A (n=1 and B1 (n=1 carrying transferable blaCTX-M-14 and the new blaTEM-153, respectively. A 80kb IncK-blaCTX-M-14 was found, being highly related to that widely spread among CTX-M-14 producers of humans and animals from Portugal and other European countries. sul genes were found in 88% (22/25;sul2-60%, sul1-48%, sul3-4% of the sulfonamide resistant isolates. Class 1 integrons were more frequently found than class 2 (7% vs 3%. Interestingly, gene cassette arrangements within these platforms were identical to those commonly observed among Enterobacteriaceae from Portuguese food-producing animals, although aadA13 is here firstly described in Morganella morganii. These results reinforce the relevance of human commensal flora as reservoir of clinically relevant antibiotic resistance genes including blaESBLs, and highly transferable genetic platforms as IncK epidemic

  16. blaCTX-M-I group extended spectrum beta lactamase-producing Salmonella typhi from hospitalized patients in Lagos, Nigeria

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    Akinyemi KO

    2015-05-01

    Full Text Available Kabiru O Akinyemi,1 Bamidele A Iwalokun,2 Olajide O Alafe,1 Sulaiman A Mudashiru,1 Christopher Fakorede,11Department of Microbiology, Lagos State University, Ojo, Lagos, Nigeria; 2Biochemistry and Nutrition Division, Nigerian Institute of Medical Research, Yaba, Lagos, NigeriaPurpose: The global spread of blaCTX-M-I extended-spectrum beta-lactamase (ESBL-producing Salmonella spp. remains a major threat to treatment and control. Evidence of emergence and spread of this marker are lacking in Nigeria. This study investigated blaCTX-M-I ESBL production among Salmonella isolates from hospitalized patients.Methods: Patients (158 total made up of two groups were evaluated. Group A was composed of 135 patients with persistent pyrexia and group B was composed of 23 gastroenteritis patients and their stool samples. Samples were cultured, and isolates were identified and were subjected to antibiotic susceptibility testing by standard methods. Isolates were further screened for ESBL production, blaCTX-M-I genes and transferability by double disk synergy test, plasmid extraction, polymerase chain reaction, and conjugation experiment.Results: Thirty-five (25.9% Salmonella isolates were identified from group A, of which 74.3% were S. typhi, 22.9% were S. paratyphi and two (5.7% were invasive non-typhoidal S. enteritidis. Nine Plasmodium falciparum infections were recorded, four of which were identified as co-infections with typhoidal Salmonella. Only two (8.7% S. enteritidis samples were obtained from group B (P>0.05. A total of 24 isolates were ESBL-positive, eliciting resistance to five to seven antibiotics, and were multiple-drug resistant. ESBL production due to the blaCTX-M-I gene cluster was detected in eleven (45.8% Salmonella isolates. Nine (81.8% of the eleven blaCTX-M-I ESBL producers were S. typhi and two (18.2% isolates were S. enteritidis. Four of nine S. typhi blaCTX-M-I ESBL-producing strains harbored 23 kb self-transmissible plasmid that was co

  17. Emergence of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae in the Central African Republic: genetic characterization

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    Frank Thierry

    2011-08-01

    Full Text Available Abstract Background Cross-resistance to quinolones and beta-lactams is frequent in Enterobacteriaceae, due to the wide use of these antibiotics clinically and in the food industry. Prescription of one of these categories of antibiotic may consequently select for bacteria resistant to both categories. Genetic mechanisms of resistance may be secondary to a chromosomal mutation located in quinolone resistance determining region of DNA gyrase or topoisomerase IV or to a plasmid acquisition. The insertion sequence ISCR1 is often associated with qnr and may favour its dissemination in Gram-negative bacteria. The aim of this study was to determine the genetic mechanism of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in the Central African Republic. Findings Among seventeen ESBL-producing Enterobacteriaceae isolated from urine, pus or stool between January 2003 and October 2005 in the Central African Republic, nine were resistant to ciprofloxacin (seven from community patients and two from hospitalized patients. The ESBL were previously characterized as CTX-M-15 and SHV-12. Susceptibility to nalidixic acid, norfloxacin and ciprofloxacin, and the minimal inhibitory concentrations of these drugs were determined by disc diffusion and agar dilution methods, respectively. The presence of plasmid-borne ISCR1-qnrA region was determined by PCR and amplicons, if any, were sent for sequencing. Quinolone resistance determining region of DNA gyrase gyrA gene was amplified by PCR and then sequenced for mutation characterization. We found that all CTX-M-producing strains were resistant to the tested quinolones. All the isolates had the same nucleotide mutation at codon 83 of gyrA. Two Escherichia coli strains with the highest MICs were shown to harbour an ISCR1-qnrA1 sequence. This genetic association might favour dissemination of resistance to quinolone and perhaps other antibiotics among Enterobacteriaceae

  18. Extended-spectrum beta-lactamases among Enterobacteriaceae isolated in a public hospital in Brazil Beta-lactamases de espectro estendido em Enterobacteriaceae isoladas de Hospital Público no Brasil

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    Milena Dropa

    2009-08-01

    Full Text Available Extended-spectrum β-lactamases (ESBL in enterobacteria are recognized worldwide as a great hospital problem. In this study, 127 ESBL-producing Enterobacteriaceae isolated in one year from inpatients and outpatients at a public teaching hospital at São Paulo, Brazil, were submitted to analysis by PCR with specific primers for blaSHV, blaTEM and blaCTX-M genes. From the 127 isolates, 96 (75.6% Klebsiella pneumoniae, 12 (9.3% Escherichia coli, 8 (6.2% Morganella morganii, 3 (2.3% Proteus mirabilis, 2 (1.6% Klebsiella oxytoca, 2 (1.6% Providencia rettgeri, 2 (1.6% Providencia stuartti, 1 (0.8% Enterobacter aerogenes and 1 (0.8% Enterobacter cloacae were identified as ESBL producers. BlaSHV, blaTEM and blaCTX-M were detected in 63%, 17.3% and 33.9% strains, respectively. Pulsed field gel eletrophoresis genotyping of K. pneumoniae revealed four main molecular patterns and 29 unrelated profiles. PCR results showed a high variety of ESBL groups among strains, in nine different species. The results suggest the spread of resistance genes among genetically different strains of ESBL-producing K. pneumoniae in some hospital wards, and also that some strongly related strains were identified in different hospital wards, suggesting clonal spread in the institutional environment.Beta-lactamases de espectro estendido (ESBL em enterobactérias são reconhecidas mundialmente como um grande problema hospitalar. Neste estudo, 127 Enterobacteriaceae produtoras de ESBL isoladas por um ano, de pacientes internados e ambulatoriais de um hospital público de ensino em São Paulo, Brasil, foram submetidas à análise pela PCR com iniciadores específicos para os genes blaSHV, blaTEM e blaCTX-M. Dos 127 isolados, 96 (75,6% K. pneumoniae, 12 (9,3% E. coli, 8 (6,2% M. morganii, 3 (2,3% Proteus mirabilis, 2 (1,6% Klebsiella oxytoca, 2 (1,6% Providencia rettgeri, 2 (1,6% Providencia stuartti, 1 (0,8% Enterobacter aerogenes e 1 (0,8% Enterobacter cloacae foram identificados

  19. Genomic Dissection of Travel-Associated Extended-Spectrum-Beta-Lactamase-Producing Salmonella enterica Serovar Typhi Isolates Originating from the Philippines: a One-Off Occurrence or a Threat to Effective Treatment of Typhoid Fever?

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas; Mikoleit, Matthew;

    2015-01-01

    One unreported case of extended-spectrum-beta-lactamase (ESBL)-producing Salmonella enterica serovar Typhi was identified, whole-genome sequence typed, among other analyses, and compared to other available genomes of S. Typhi. The reported strain was similar to a previously published strain harbo...

  20. SYNTHESIS AND PROCESSING OF ESCHERICHIA-COLI TEM-BETA-LACTAMASE AND BACILLUS-LICHENIFORMIS ALPHA-AMYLASE IN ESCHERICHIA-COLI : THE ROLE OF SIGNAL PEPTIDASE-I

    NARCIS (Netherlands)

    van Dijl, J M; SMITH, H; BRON, S; VENEMA, G

    1988-01-01

    A mutant of Escherichia coli, in which signal peptidase I synthesis can be regulated, was constructed. The mutant was used to study the effects of signal peptidase I limitation on the synthesis and efficiency of processing of two proteins: the periplasmic E. coli TEM-beta-lactamase and Bacillus lich

  1. Activity of sulbactam in combination with ceftriaxone in vitro and in experimental endocarditis caused by Escherichia coli producing SHV-2-like beta-lactamase.

    Science.gov (United States)

    Fantin, B; Pangon, B; Potel, G; Caron, F; Vallée, E; Vallois, J M; Mohler, J; Buré, A; Philippon, A; Carbon, C

    1990-04-01

    We studied the efficacy of sulbactam, a beta-lactamase inhibitor, in combination with ceftriaxone in vitro and in experimental endocarditis due to an Escherichia coli strain producing an extended-spectrum beta-lactamase most similar to SHV-2, a new mechanism of resistance to broad-spectrum cephalosporins among members of the family Enterobacteriaceae. In vitro, ceftriaxone demonstrated an important inoculum effect (MICs were 2 and 256 micrograms/ml with 5 X 10(5) and 5 X 10(7) CFU of inoculum per ml, respectively). Sulbactam inhibited the beta-lactamase degradation of ceftriaxone and enhanced the killing by ceftriaxone with both inocula tested. In vivo, sulbactam (100 mg/kg every 8 h) or ceftriaxone (15 or 30 mg/kg every 24 h) alone were ineffective after a 4-day therapy. The addition of sulbactam to ceftriaxone (15 mg/kg) or to the ceftriaxone (15 mg/kg)-netilmicin (6 mg/kg every 24 h) combination produced a reduction of 2 log10 CFU/g of vegetation greater than that produced by therapy without sulbactam. The sulbactam-ceftriaxone (30 mg/kg) combination produced a reduction of almost 5 log10 CFU/g of vegetation greater than that produced by single-drug therapy (P less than 0.01), sterilized five of eight vegetations (versus none of seven for ceftriaxone [30 mg/kg] alone; P less than 0.05), and was as effective as the ceftriaxone (15 mg/kg)-sulbactam-netilmicin combination. We concluded that (i) SHV-2 production was responsible for ceftriaxone failure in vivo, probably because of the high inoculum present in vegetations; (ii) sulbactam used in a regimen which provided levels in serum constantly above 4 micrograms/ml and a vegetation/serum peak ratio of approximately 1:3 enhanced the activity of a broad-spectrum cephalosporin in a severe experimental infection; and (iii) the highest dose of ceftriaxone in combination with sulbactam was as effective as the lowest dose of ceftriaxone plus sulbactam plus an aminoglycoside. PMID:2188586

  2. Nosocomial acquisition of methicillin-resistant Staphyloccocus aureus (MRSA and extended-spectrum beta-lactamase (ESBL Enterobacteriaceae in hospitalised patients: a prospective multicenter study

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    De Angelis Giulia

    2012-03-01

    Full Text Available Abstract Background The risk of acquisition of antibiotic resistant-bacteria during or shortly after antibiotic therapy is still unclear and it is often confounded by scarce data on antibiotic usage. Primary objective of the study is to compare rates of acquisition of methicillin-resistant Staphylococcus aureus and extended spectrum beta-lactamase-producing Enterobacteriaceae in hospitalised patients, after starting antibiotic therapy. Methods/Design The study, running in three European hospitals, is a multicenter, prospective, longitudinal, observational cohort study funded from the European Community's Seventh Framework Programme [FP7/2007-2013] within the project 'Impact of Specific Antibiotic Therapies on the prevalence of hUman host ResistaNt bacteria' (acronym SATURN. Nasal and rectal screening for methicillin-resistant Staphylococcus aureus and extended spectrum beta-lactamase-producing Enterobacteriaceae will be obtained at hospital admission, discharge, at antibiotic start (t0, within one hour and at the following intervals: day 3 (t1, 7 (t2, 15 (t3, and 30 (t4. Two nested case-control studies will be performed. The objective of the first study will be to define individual level of risk related to specific antibiotics. Patients acquiring methicillin-resistant Staphylococcus aureus and extended spectrum beta-lactamase-producing Enterobacteriaceae (cases will be compared with patients not acquiring antibiotic-resistant strains after starting antibiotic therapy (controls; ratio 1:4. To define the impact of antibiotics on new acquisition of target antibiotic-resistant bacteria, a second nested case-control study will be done (ratio 1:4. Control group will be selected among patients not receiving antibiotics, admitted in the same ward on the day of the corresponding case, with negative cultures at admission. Epidemiological, clinical and microbiological data will be prospective collected. Discussion The rationale of this study is to better

  3. High prevalence of extended-spectrum and plasmidic AmpC beta-lactamase-producing Escherichia coli from poultry in Tunisia.

    Science.gov (United States)

    Maamar, Elaa; Hammami, Samia; Alonso, Carla Andrea; Dakhli, Nouha; Abbassi, Mohamed Salah; Ferjani, Sana; Hamzaoui, Zaineb; Saidani, Mabrouka; Torres, Carmen; Boutiba-Ben Boubaker, Ilhem

    2016-08-16

    This study was conducted to detect extended spectrum beta-lactamases (ESBLs) and plasmidic AmpC beta-lactamase (pAmpC-BL)-producing Escherichia coli isolates in industrial poultry samples were collected from healthy chickens of the three farms. Samples were inoculated onto desoxycholate-lactose-agar plates supplemented with cefotaxime (2mg/L). E. coli was identified by biochemical and molecular methods and antibiotic susceptibility testing by the disk diffusion method. Genes encoding ESBLs and pAmpC-BL were detected by PCR and sequencing. Phylogenetic groups were determined by triplex PCR. The molecular typing of strains was done by pulsed field gel electrophoresis (PFGE) and Multilocus Sequence Typing (MLST) in those isolates showing different PFGE patterns. Cefotaxime-resistant E. coli isolates were recovered in 48 of 137 fecal samples (35%), and one isolate/sample was further studied. The following beta-lactamase genes were detected: blaCTX-M-1 (29 isolates, isolated in all three farms), blaCTX-M-15 (5 isolates, confined in farm II), blaCTX-M-14 and blaCMY-2 (one isolate and 13 isolates, respectively, in farm III). The 48 cefotaxime-resistant isolates were distributed into phylogroups: B1 (n=21), A (n=15) and D (n=12). PFGE analysis revealed 19 unrelated patterns: 15 different profiles among ESBL-positive strains and 4 among the CMY-2-positive isolates. The following sequence types-associated phylogroups were detected: a) CTX-M-1-positive strains: lineages ST542-B1, ST212-B1, ST58-B1, ST155-B1 and ST349-D; b) CTX-M-15-positive strain: lineage ST405-D; c) CTX-M-14-positive strain: lineage ST1056-B1; d) CMY-2-positive strains: lineages ST117-D, ST2197-A, and ST155-B1. Healthy chickens constitute an important reservoir of ESBL- and pAmpC-BL-producing E. coli isolates that potentially could be transmitted to humans via the food chain or by direct contact. PMID:27220012

  4. Crystal Structures of KPC-2[beta]-Lactamase in Complex with 3-Nitrophenyl Boronic Acid and the Penam Sulfone PSR-3-226

    Energy Technology Data Exchange (ETDEWEB)

    Ke, Wei; Bethel, Christopher R.; Papp-Wallace, Krisztina M.; Pagadala, Sundar Ram Reddy; Nottingham, Micheal; Fernandez, Daniel; Buynak, John D.; Bonomo, Robert A.; van den Akker, Focco (Case Western); (Stokes); (SMU)

    2012-08-01

    Class A carbapenemases are a major threat to the potency of carbapenem antibiotics. A widespread carbapenemase, KPC-2, is not easily inhibited by {beta}-lactamase inhibitors (i.e., clavulanic acid, sulbactam, and tazobactam). To explore different mechanisms of inhibition of KPC-2, we determined the crystal structures of KPC-2 with two {beta}-lactamase inhibitors that follow different inactivation pathways and kinetics. The first complex is that of a small boronic acid compound, 3-nitrophenyl boronic acid (3-NPBA), bound to KPC-2 with 1.62-{angstrom} resolution. 3-NPBA demonstrated a Km value of 1.0 {+-} 0.1 {micro}M (mean {+-} standard error) for KPC-2 and blocks the active site by making a reversible covalent interaction with the catalytic S70 residue. The two boron hydroxyl atoms of 3-NPBA are positioned in the oxyanion hole and the deacylation water pocket, respectively. In addition, the aromatic ring of 3-NPBA provides an edge-to-face interaction with W105 in the active site. The structure of KPC-2 with the penam sulfone PSR-3-226 was determined at 1.26-{angstrom} resolution. PSR-3-226 displayed a K{sub m} value of 3.8 {+-} 0.4 {micro}M for KPC-2, and the inactivation rate constant (kinact) was 0.034 {+-} 0.003 s{sup -1}. When covalently bound to S70, PSR-3-226 forms a trans-enamine intermediate in the KPC-2 active site. The predominant active site interactions are generated via the carbonyl oxygen, which resides in the oxyanion hole, and the carboxyl moiety of PSR-3-226, which interacts with N132, N170, and E166. 3-NPBA and PSR-3-226 are the first {beta}-lactamase inhibitors to be trapped as an acyl-enzyme complex with KPC-2. The structural and inhibitory insights gained here could aid in the design of potent KPC-2 inhibitors.

  5. Surto em berçário por Klebsiella pneumoniae produtora de beta-lactamase de espectro estendido atribuído à colonização de profissional de saúde portador de onicomicose Outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in an intermediate-risk neonatal unit linked to onychomycosis in a healthcare worker

    Directory of Open Access Journals (Sweden)

    Valéria C. Cassettari

    2006-08-01

    Full Text Available OBJETIVO: Descrever surto por Klebsiella pneumoniae produtora de beta-lactamase de espectro estendido em berçário de risco intermediário. MÉTODOS: Após identificação dos primeiros casos, a situação foi conduzida como surto, sendo intensificadas as medidas básicas de prevenção de infecções hospitalares e investigadas possíveis fontes de disseminação da bactéria. RESULTADOS: O surto durou 6 meses e atingiu 36 recém-nascidos, causando sete infecções e 29 colonizações. Na primeira fase do surto, os portadores evoluíram com infecção, porém, na segunda fase, os portadores eram assintomáticos e só foram identificados por culturas de vigilância. O surto foi resolvido após identificação e tratamento de profissional de saúde que apresentava onicomicose e era portadora de Klebsiella pneumoniae produtora de beta-lactamase de espectro estendido nas mãos. CONCLUSÃO: Detecção e controle da disseminação oculta da bactéria multirresistente entre os recém-nascidos de menor risco evitou sua instalação endêmica no berçário, bem como a conseqüente exposição dos pacientes mais graves e suscetíveis à infecção.OBJECTIVE: To describe an outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in an intermediate-risk neonatal unit. METHODS: After the identification of the first cases, the situation was regarded as an outbreak, and basic preventive measures against nosocomial infections were strictly enforced, and possible sources of dissemination were investigated. RESULTS: The outbreak lasted for 6 months and affected 36 newborn infants, causing seven infections and 29 colonizations. In the first stage of the outbreak, patients developed infection, but in the second stage, they were asymptomatic and were only identified by surveillance cultures. The outbreak was controlled after the identification and treatment of the healthcare worker who had been diagnosed with onychomycosis and whose hands were

  6. Estudo da produção de beta -lactamase e sensibilidade às drogas em linhagens de estafilococos coagulase-negativos isolados de recém-nascidos Study of production of beta-lactamase and drugs susceptibility in strains of coagulase-negative staphylococci isolated of neonates

    Directory of Open Access Journals (Sweden)

    Maria de Lourdes Ribeiro de Souza da Cunha

    2002-01-01

    Full Text Available Os estafilococos coagulase-negativos (ECN, embora reconhecidos como saprófitas por muito tempo, têm emergido como agentes etiológicos de uma série de infecções, sendo atualmente os principais responsáveis por sepse em UTI neonatal. Tendo em vista estas características, este estudo objetivou a identificação de estafilococos coagulase-negativos isolados de processos infecciosos em recém-nascidos, bem como a determinação da produção de beta-lactamase e sensibilidade às drogas pelas linhagens isoladas. O Staphylococcus epidermidis foi a espécie mais freqüentemente isolada (77,8%. O estudo da produção de beta-lactamase revelou esta característica na maioria das linhagens de ECN isoladas (71,8%. As linhagens de ECN mostraram, ainda, resistência múltipla aos antibióticos utilizados, com 63,2% dos isolados apresentando resistência a cinco ou mais drogas. A elevada transmissibilidade de plasmídios entre estas linhagens e o uso abusivo de drogas antimicrobianas têm-se constituído em importantes fatores na seleção de amostras multirresistentes e na transferência de genes de resistência.Although coagulase-negative staphylococci (CNS have been recognized as saprophytes for a long time, they had emerged as etiologic agents of infections. They have currently been the most frequently isolated pathogen in sepsis in neonatal intensive care unit (NICU. This study aimed the identification of CNS strains isolated from newborns' infections and to determination of beta-lactamase and drugs susceptibility. Staphylococcus epidermidis was the most frequently isolated species (77,8%. The study of the beta-lactamase production revealed this characteristic in the most of the strains of CNS isolated (71,8%. The strains isolated in this study presented multiple resistance to the antibiotics tested, with 63,2% of isolates presenting resistance to five or more drugs. The high transmissibility of plasmids among those strains and the abusive use of

  7. Immobilization of the enzyme {beta}-lactamase by self-assembly on thin films of a poly(phenyleneethynylene) sequenced with flexible segments containing sulfur atoms

    Energy Technology Data Exchange (ETDEWEB)

    Vazquez, Erika [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Aguilar, Abdieel Esquivel [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Facultad de Ciencias Quimicas, Universidad Autonoma de Coahuila, Blvd. V. Carranza and Ing. J. Cardenas, 25000 Saltillo (Mexico); Moggio, Ivana [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico)]. E-mail: imoggio@ciqa.mx; Arias, Eduardo [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Romero, Jorge [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Barrientos, Hector [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Torres, Jose Roman [Centro de Investigacion en Quimica Aplicada (CIQA), Blvd. Enrique Reyna 140, 25253, Saltillo (Mexico); Luz Reyes Vega, Maria de la [Facultad de Ciencias Quimicas, Universidad Autonoma de Coahuila, Blvd. V. Carranza and Ing. J. Cardenas, 25000 Saltillo (Mexico)

    2007-05-16

    A novel poly(phenyleneethynylene) sequenced in the main conjugated chain with flexible groups containing sulfur atoms has been synthesized by Heck-Sonogashira coupling reaction. Layer-by-layer films of the polymer have been prepared with a linear growth in thickness up to four layers as evidenced by UV-Vis spectroscopy and perfilometry. On the top of these multilayers, the enzyme {beta}-lactamase was deposited by self-assembly. The enzymatic activity was measured by a modified spectrophotometric standard assay method for penicillin G, ampicillin and amoxicillin. A higher and faster activity was obtained for penicillin G and thus preliminary study of the biosensor response by fluorescence was carried out for this antibiotic revealing a decrease in the polymer fluorescence as function of the penicillin G concentration.

  8. Expression, purification, crystallization, and preliminary X-ray crystallographic analysis of OXA-17, an extended-spectrum {beta}-lactamase conferring severe antibiotic resistance

    Energy Technology Data Exchange (ETDEWEB)

    Lee, J. H., E-mail: msgjhlee@mju.ac.kr; Sohn, S. G., E-mail: sgsohn@mju.ac.kr; Jung, H. I., E-mail: jhinumber1@hanmail.net; An, Y. J., E-mail: anyj0120@hanmail.net; Lee, S. H., E-mail: sangheelee@mju.ac.kr [Myongji University, Drug Resistance Proteomics Laboratory, Department of Biological Sciences (Korea, Republic of)

    2013-07-15

    OXA-17, an extended-spectrum {beta}-lactamase (ESBL) conferring severe antibiotic resistance, hydrolytically inactivates {beta}-lactam antibiotics, inducing a lack of eradication of pathogenic bacteria by oxyimino {beta}-lactams and not helping hospital infection control. Thus, the enzyme is a potential target for developing antimicrobial agents against pathogens producing ESBLs. OXA-17 was purified and crystallized at 298 K. X-ray diffraction data from OXA-17 crystal have been collected to 1.85 A resolution using synchrotron radiation. The crystal of OXA-17 belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 48.37, b = 101.12, and c = 126.07 A. Analysis of the packing density shows that the asymmetric unit probably contains two molecules with a solvent content of 54.6%.

  9. Presence of blaPER-1 and blaVEB-1 beta-lactamase genes among isolates of Pseudomonas aeruginosa from South West of Iran.

    Science.gov (United States)

    Davodian, Elham; Sadeghifard, Nourkhoda; Ghasemian, Abdolmajid; Noorbakhsh, Samileh

    2016-09-01

    Pseudomonas aeruginosa isolates have acquired resistance to antibiotics such as novel beta-lactams. The aim of this study was to investigate the blaPER-1, blaVEB-1, and blaPSE-1 genes among isolates of P. aeruginosa among intensive care unit (ICU) patients. Sixty-five isolates were collected. The antibiotic susceptibility testing and combined disk tests were performed to detect the isolates producing extended spectrum beta-lactamases (ESBLs) among ceftazidime-resistant isolates. Polymerase chain reaction (PCR) amplification of blaPER-1, blaVEB-1, and blaPSE-1 genes was conducted. Ten (15.3%) isolates were ESBL-positive, of which 40% (n=4) belonged to males and 60% (n=6) were collected from females. Moreover, two and one isolates harbored blaPER-1 and blaVEB-1 genes, respectively. PMID:26944896

  10. Plasmid-mediated extended-spectrum beta-lactamase-producing strains of Enterobacteriaceae isolated from diabetes foot infections in a Brazilian diabetic center

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    R.N. Motta

    2003-04-01

    Full Text Available We bacteriologically analyzed 156 species of Enterobacteriaceae, isolated from 138 patients with community-acquired diabetic foot ulcers, in a prospective study made at a diabetic center and at the Federal University of Ceará, Brazil, from March, 2000, to November, 2001.The samples were cultured using selective media, and identification, susceptibility tests and detection of plasmid-mediated-extended-spectrum-beta-lactamase (ESBL producing strains were made with conventional and automated methods. The most frequently occurring pathogens were K. pneumoniae (21.2%, Morganella morganii (19.9% and E. coli (15.4%. High resistance rates were noted for ampicillin, first generation cephalosporin, trimethoprim/sulfamethoxazole, tetracycline, amoxicillin-clavulanic acid and chloramphenicol. ESBL-producing strains were detected in 6% of the patients. Resistance among gram-negative bacteria has become increasingly common, even in community-acquired infections.

  11. Investigations of multiresistance to antibiotics and chemotherapeutics and extended spectrum beta: Lactamase effect (ESBL test in strains E.coli and salmonella originating from domestic animals

    Directory of Open Access Journals (Sweden)

    Mišić Dušan

    2006-01-01

    Full Text Available The presence of multiresistance to the effects of antibiotics and chemotherapeutics and extended spectrum beta-lactamase were examined in 45 strains of E. coli and 35 strains of Salmonella. The strains of E. coli originated from several species of domestic animals: dogs, cats, poultry, and cattle, and 30 strains of Salmonella originated from poultry, 4 strains from cattle, and 1 strain from swine. The presence of the following serovarieties was established using serological examinations: Salmonella Enteritidis 17 strains, Salmonella Gallinarum 1 strain, Salmonella Hartford 5 strains, Salmonella Anatum 1 strain, Salmonella Typhimurium 4 strains, Salmonella Agona 1 strain, Salmonella Infantis 1 strain, Salmonella Thompson var. Berlin 1 strain, Salmonella Tennessee 1 strain, Salmonella Senftenberg 1 strain, Salmonella Glostrup 1 strain, and Salmonella Hadar 1 strain. In the examinations of the listed strains we used antibiogram discs of ampicillin, amoxicillin with clavulanic acid, cephalexin, cephtriaxon, cephotaxim, cephtazidime, aztreonam, gentamycin, chloramphenicol, tetracycline, cyprofloxacine, and a combination of sulphamethoxasole and trimethoprim. The lowest prevalence of multiresistance in E. Coli strains to 3 or more antibiotics was established in dogs 20%, and the highest in 60% strains originating from swine. In 62.88% strains of Salmonella we established sensitivity to all applied antibiotics. Resistance was also established in a small number of the examined strains to ampicillin (11 strains, to tetracycline (5 strains, to amoxicillin with clavulanic acid (5 strains, to sulphamethoxasole with trimethoprim (5 strains, to gentamycin (3 strains, and to cloramphenicol (1 strain. Of all the examined strains of Salmonella, 6 strains originating from poultry exhibited multiresistence. The presence of extended spectrum beta-lactamase effects examined using the ESBL test, was not established in strains of E. coli and Salmonella strains.

  12. Use of ampicillin-sulbactam for treatment of experimental meningitis caused by a beta-lactamase-producing strain of Escherichia coli K-1.

    Science.gov (United States)

    Guerra-Romero, L; Kennedy, S L; Fournier, M A; Tureen, J H; Täuber, M G

    1991-10-01

    We evaluated the pharmacokinetics and therapeutic efficacy of ampicillin combined with sulbactam in a rabbit model of meningitis due to a beta-lactamase-producing strain of Escherichia coli K-1. Ceftriaxone was used as a comparison drug. The MIC and MBC were 32 and greater than 64 micrograms/ml (ampicillin), greater than 256 and greater than 256 micrograms/ml (sulbactam), 2.0 and 4.0 micrograms/ml (ampicillin-sulbactam [2:1 ratio, ampicillin concentration]) and 0.125 and 0.25 micrograms/ml (ceftriaxone). All antibiotics were given by intravenous bolus injection in a number of dosing regimens. Ampicillin and sulbactam achieved high concentrations in cerebrospinal fluid (CSF) with higher dose regimens, but only moderate bactericidal activity compared with that of ceftriaxone was obtained. CSF bacterial titers were reduced by 0.6 +/- 0.3 log10 CFU/ml/h with the highest ampicillin-sulbactam dose used (500 and 500 mg/kg of body weight, two doses). This was similar to the bactericidal activity achieved by low-dose ceftriaxone (10 mg/kg), while a higher ceftriaxone dose (100 mg/kg) produced a significant increase in bactericidal activity (1.1 +/- 0.4 log10 CFU/ml/h). It appears that ampicillin-sulbactam, despite favorable CSF pharmacokinetics in animals with meningitis, may be of limited value in the treatment of difficult-to-treat beta-lactamase-producing bacteria, against which the combination shows only moderate in vitro activity. PMID:1759824

  13. Inhibitory effects of various essential oils and individual components against extended-spectrum beta-lactamase (ESBL) produced by Klebsiella pneumoniae and their chemical compositions.

    Science.gov (United States)

    Orhan, Ilkay Erdogan; Ozcelik, Berrin; Kan, Yüksel; Kartal, Murat

    2011-10-01

    In the current study, in vitro inhibitory activity of several essential oils obtained from the cultivated plants, Foeniculum vulgare, Mentha piperita and M. spicata, Ocimum basilicum, Origanum majorana, O. onites, O. vulgare, Satureja cuneifolia, and a number of individual essential oil components of terpene and aromatic types were screened against 10 isolated strains of Klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) enzyme, which makes this microorganism quite resistant against the antibiotics: trimetoprime-sulfametoksazol, sulbactam-ampicilin, clavulonate-amoxicilin, ceftriaxon, cefepime, imipenem, ceftazidime, tobramicine, gentamisine, ofloxacin, and ciprofloksasin. All of the essential oils and the components exerted a remarkable inhibition ranging between 32 and 64 μg/mL against all of these strains as strong as the references (ampicilin and oflaxocin) inhibiting at 32 μg/mL. Besides, chemical compositions of the essential oils were elucidated by gas chromatography-mass spectrometry (GC-MS). The essential oils and the pure components widely found in essential oils screened herein have shown remarkable inhibition against ESBL-producing K. pneumoniae strains, which leads to the suggestion that they may be used as food preservatives for this purpose. Practical Application:  The essential oils obtained from Foeniculum vulgare, Mentha piperita and M. spicata, O.cimum basilicum, Origanum majorana, O. onites, O. vulgare, and Satureja cuneifolia as well as common essential oil components have shown notable inhibitory effects against 10 isolated strains of Klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) enzyme and they might be used as food preservative or ingredient. PMID:22417594

  14. Multidrug resistant NDM-1 metallo-beta-lactamase producing Klebsiella pneumoniae sepsis outbreak in a neonatal intensive care unit in a tertiary care center at central India

    Directory of Open Access Journals (Sweden)

    Atul Khajuria

    2014-01-01

    Full Text Available Objective: The objective of the following study is to detect genes encoding carbapenem resistance in Klebsiella pneumoniae sepsis outbreak in a neonatal intensive care unit (NICU. Materials and Methods: Antibiotic sensitivity test was performed by standard Kirby Bauer disc diffusion technique and minimum inhibitory concentrations of antibiotics was determined by VITEK-2. Polymerase chain reaction (PCR assays and sequencing was used to determine the presence of beta-lactamase encoding genes. Conjugation experiments were performed to determine the transferability of beta-lactamase. Isolate relatedness were determined by repetitive-element PCR (REP, enterobacterial repetitive intergenic consensus (ERIC PCR and random amplified polymorphic deoxyribonucleic acid (RAPD. Results: All the isolates were completely resistant to the second and third generation cephalosporins tested as well as carbapenems. Susceptibility profiling of the isolates indicated that 100% retained susceptibility to tigecycline and colistin. Conjugation experiments indicated that blaNDM-1 was transferable and likely through a plasmid-mediated event. All the isolates showed the presence of blaNDM-1 with co association of bla CTX-M-15 . REP-PCR, ERIC-PCR and RAPD revealed a single clonal type circulating in NICU environment. Conclusion: Co-production of NDM-1 with CTX-M-15 in K. pneumoniae isolates was detected for the first time in our NICU. Transmission of plasmid carrying these resistant genes to other members of Enterobacteriaceae will increase the incidence of multidrug resistance. Early detection of these genes will help in prevention and adequate infection control by limiting the spread of these organisms.

  15. Prevalência de produção de beta-lactamases de espectro estendido em bacteremias por Klebsiella pneumoniae e Escherichia coli

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    Zavascki, Alexandre Prehn

    2007-01-01

    Full Text Available Objetivos: avaliar a prevalência da produção de beta-lactamases de espectro estendido (extendedspectrum beta-lactamases – ESBL por Klebsiella pneumoniae e Escherichia coli isoladas de pacientes adultos hospitalizados e descrever as características clínicas dos pacientes. Métodos: em estudo transversal retrospectivo, foram revisados os prontuários de pacientes adultos que apresentaram bacteremia por Klebsiella pneumoniae e Escherichia coli no Hospital São Lucas da PUCRS, no período de junho de 2004 a março de 2006. Resultados: um total de 145 pacientes foram incluídos no estudo; destes, em 51 (35,2% foi isolada uma bactéria produtora de ESBL; a prevalência de ESBL foi maior em Klebsiella pneumoniae (45 isolados, 55,6% do que em Escherichia coli (6 isolados, 9,4% p<0,001. O uso de antimicrobianos beta-lactâmicos dentro dos 14 dias prévios ao isolamento foi mais comum no grupo ESBL (50 pacientes, 98% que no grupo não-ESBL (15 pacientes, 16%; p<0,001. O imipenem foi a droga com maior taxa de eficácia in vitro contra isolados produtores de ESBL (100%. Conclusões: o estudo demonstrou uma elevada prevalência de produção de ESBL em bacteremias por K. pneumoniae. Pacientes com bacteremia por isolados produtores de ESBL apresentaram maior mortalidade e o uso prévio de beta-lactâmicos esteve fortemente associado à produção dessa enzima. É necessária a vigilância contínua da prevalência de ESBL em enterobactérias, para definição de esquemas terapêuticos e políticas de controle de infecção.

  16. Molecular Study of Extended-Spectrum Beta-Lactamases among Enterobacter spp. Isolated from Teaching Hospitals in Tehran and Qazvin, Iran

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    Amir Peymani

    2013-10-01

    Full Text Available Abstract Background and objective: Enterobacter spp. are members of the family Enterobacteriaceae. In recent years, there are several reports showing the high prevalence of beta-lactam resistant among these organisms. Different types of beta-lactamase enzymes have been identified in Enterobacter spp., which Extended-Spectrum Beta-Lactamases (ESBLs are very important among them. The ESBL-producing isolates capable to hydrolyze extended spectrum penicillin's and cephalosporins. The aims of this study were to determine the prevalence of ESBLs and to detect of blaTEM and blaSHV genes among ESBL-producing Enterobacter spp. isolates. Materials and methods: A total of 107 Enterobacter isolates were collected from Tehran and Qazvin hospitals. Isolates were identified by standard microbiology and biochemical methods. All isolates were screened for ESBLs production by agar disk diffusion method using ceftazidime, cefotaxime, aztreonam, cefpodoxime and ceftriaxone discs and then were confirmed by combined disk method. ESBLs-positive isolates were tested for blaTEM and blaSHV-ESBLs genes. Results: Of the 107 isolates, 58 isolates (54.2% showed the reduced susceptibility to the screening antibiotics. 51 isolates (47.6% were ESBLs positive by combined disks method. Twenty eight isolates (54.9% contained blaTEM genes and 2 isolates (3.9% harbored the blaSHV genes. Conclusions: This study showed the high prevalence of ESBLs in Enterobacter spp. isolated from the clinical specimens in studied hospitals. The initial identification of ESBL-producing isolates and use of appropriate infection strategy is essential to control the more spread of these organisms in selected hospitals.

  17. Metallo-beta-lactamase producing Pseudomonas aeruginosa strains isolated in hospitals in Recife, PE, Brazil Produção de metalo-beta-lactase de linhagens de Pseudomonas aeruginosa isoladas em hospitais do Recife, PE, Brasil

    Directory of Open Access Journals (Sweden)

    Vera Magalhães

    2005-06-01

    Full Text Available Out of 24 nosocomial strains of Pseudomonas aeruginosa from Recife, Brazil, 15 (62% were metallo-beta-lactamase producers. Such isolates were resistant to main antipseudomonas drugs, except polymixyn B and aztreonam. The enzyme responsible for the carbapenem-resistance belongs to SPM-1 class, and the gene involved, blaspm-1, is likely plasmid located.De 24 linhagens hospitalares de Pseudomonas aeruginosa provenientes de Recife, Brasil, 15 (62% produziram metalo-beta-lactamase. Tais isolados foram resistentes às principais drogas antipseudomonas, exceto polimixina B e aztreonam. A enzima responsável pela resistência aos carbapanêmicos pertence à classe SPM-1 e o gene envolvido, blaspm-1, provavelmente é plasmidial.

  18. Abattoirs as Non-Hospital Source of Extended Spectrum Beta Lactamase Producers: Confirmed by the Double Disc Synergy Test and Characterized by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry

    OpenAIRE

    Ikegbunam, Moses Nkechukwu; Anagu, Linda Onyeka; Ifeanyi R Iroha; Ejikeugwu, Chika Ebiye; Esimone, Charles Okey

    2014-01-01

    In this study, the presence of extended spectrum beta lactamase (ESBL) producing organisms in abattoirs, a non-hospital community was investigated. The presence of ESBL-producing phenotypes was confirmed by the Double Disc Synergy Test (DDST). Out of the 99 isolates screened for ESBL, 28 (28.3%) were confirmed positive. The positive isolates were characterised by using Matrix-Assisted Laser Desorption/Ionization Time of flight Mass Spectrometry. 50% of the isolates were Pseudomonas spp., the ...

  19. Comparison of the inoculum effects of members of the family Enterobacteriaceae on cefoxitin and other cephalosporins, beta-lactamase inhibitor combinations, and the penicillin-derived components of these combinations.

    OpenAIRE

    Goldstein, E J; Citron, D M; Cherubin, C E

    1991-01-01

    We compared the inoculum effects of 105 recent clinical isolates of the family Enterobacteriaceae on cefoxitin, other cephalosporins, aztreonam, and three beta-lactamase inhibitors (clavulanic acid, sulbactam, and tazobactam) and their penicillin-derived components. Piperacillin and aztreonam showed the largest inoculum effect, and cefoxitin showed the smallest. The other cephalosporins tested (cefotetan, ceftizoxime, and ceftriaxone) showed an intermediate inoculum effect. In general, the in...

  20. Prevalence of 16S rRNA methylase, modifying enzyme, and extended-spectrum beta-lactamase genes among Acinetobacter baumannii isolates.

    Science.gov (United States)

    Liu, Zhenru; Ling, Baodong; Zhou, Liming

    2015-08-01

    Multidrug-resistant Acinetobacter baumannii has become a worldwide problem, and methylation of 16S rRNA has recently emerged as a new mechanism of resistance to aminoglycosides, which is mediated by a newly recognized group of 16S rRNA methylases. 16S rRNA methylase confers a high-level resistance to all 4,6-substituted deoxystreptamine aminoglycosides that are currently used in clinical practice. Some of the A. baumannii isolates have been found to coproduce extended-spectrum beta-lactamases (ESBLs), contributing to their multidrug resistance. The aim of this study was to detect the determinants of the 16S rRNA methylase genes armA, rmtA, rmtB, rmtC, rmtD, rmtE, and npmA, the modifying enzyme genes aac(6')-Ib, ant(3″)-Ia, aph(3')-I, and the extended-spectrum beta-lactamase genes bla(TEM), bla(SHV), and bla(CTX-M-3) among A. baumannii isolates in northeastern Sichuan, China. Minimum inhibitory concentrations (MICs) of 21 different antimicrobial agents against the A. baumannii isolates were determined. The clinical isolates showed a high level of resistance (MIC≧256 μg/ml) to aminoglycosides, which ranged from 50·1 to 83·8%. The resistances to meropenem and imipenem, two of the beta-lactam antibiotics and the most active antibiotics against A. baumannii, were 9·1 and 8·2%, respectively. Among 60 amikacin-resistant isolates, only the 16S rRNA methylase gene armA was found to be prevalent (66·7%), but the other 16S rRNA methylase genes rmtA, rmtB, rmtC, rmtD, rmtE, and npmA were not detected. The prevalences of the modifying enzyme genes aac (6')-Ib, ant (3″)-Ia, and aph (3')-I were 51·7, 81·7, and 58·3%, respectively, which are different from a previous study in which the occurrences of these genes were 3, 64, and 72%, respectively. Among the 40 isolates that were armA-positive, the prevalences of bla(TEM), bla(SHV), and bla(CTX-M-3) genes were detected for the first time in China, and their occurrences were 45, 65, and 52·5%, respectively. In all, A

  1. Prevalence and characteristics of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae isolated from rural well water in Taian, China, 2014.

    Science.gov (United States)

    Zhang, Hongna; Zhou, Yufa; Guo, Shuyuan; Chang, Weishan

    2015-08-01

    The production of extended-spectrum beta-lactamase (ESBL) is one of the major antimicrobial resistance mechanisms in Enterobacteriaceae, and the increasing number of ESBL-producing Enterobacteriaceae isolated from water environments has posed a serious threat to the public health. The study aimed to analyze prevalence and characterization of ESBL-producing Enterobacteriaceae from rural well waters in Taian, China. A total of 10 isolates expressing an ESBL phenotype, including 9 Escherichia coli (E. coli) and 1 Klebsiella pneumoniae (K. pneumoniae) was obtained from 4 (4%) out of the 100 sampled wells. ESBL genotype revealed that 9 expressed CTX-M-15 and 1 produced CTX-M-27. Five out of 8 ESBL-producing E. coli expressing CTX-M-15 belonged to ST10, which are mostly detected from human feces in China. Importantly, the only strain of CTX-M-27-producing E. coli belonged to multi-locus sequence type B2:131 (ST131), which may be related with severe infection in humans and animals. PMID:25821088

  2. An amperometric penicillin biosensor with enhanced sensitivity based on co-immobilization of carbon nanotubes, hematein, and {beta}-lactamase on glassy carbon electrode

    Energy Technology Data Exchange (ETDEWEB)

    Chen Bi; Ma Ming [Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081 (China); Su Xiaoli, E-mail: xsu@hunnu.edu.cn [Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha 410081 (China)

    2010-07-26

    An amperometric penicillin biosensor with enhanced sensitivity was successfully developed by co-immobilization of multi-walled carbon nanotubes (MWCNTs), hematein, and {beta}-lactamase on glassy carbon electrode using a layer-by-layer assembly technique. Under catalysis of the immobilized enzyme, penicillin was hydrolyzed, decreasing the local pH. The pH change was monitored amperometrically with hematein as a pH-sensitive redox probe. MWCNTs were used as an electron transfer enhancer as well as an efficient immobilization matrix for the sensitivity enhancement. The effects of immobilization procedure, working potential, enzyme quantity, buffer concentration, and sample matrix were investigated. The biosensor offered a minimum detection limit of 50 nM (19 {mu}g L{sup -1}) for penicillin V, lower than those of the conventional pH change-based biosensors by more than two orders of magnitude. The electrode-to-electrode variation of the response sensitivity was 7.0% RSD.

  3. Increasing prevalence of imipenem-resistant Pseudomonas aeruginosa and molecular typing of metallo-beta-lactamase producers in a Korean hospital.

    Science.gov (United States)

    Kim, In-Suk; Lee, Nam Yong; Ki, Chang-Seok; Oh, Won Sup; Peck, Kyong Ran; Song, Jae-Hoon

    2005-01-01

    The types of metallo-beta-lactamases (MBLs), integrons, and genetic relatedness among Pseudomonas aeruginosa were investigated with a recent high prevalence of imipenem resistance in a Korean hospital. During 2000-2003, a total of 116 non-duplicate imipenem-resistant P. aeruginosa isolates were analyzed by PCR and DNA sequencing to detect of bla (IMP-1), bla (VIM-1), bla (VIM-2), bla (SPM-1), intI 1, intI 2, and intI 3 genes. Among them, MBL-producing isolates were evaluated for genetic relatedness using pulsed-field gel electrophoresis (PFGE) profiles. Of 116 isolates, 21 (18.1%) carried bla (VIM-2) gene with the intI 1 gene. Analysis of VIM-2 procuders by PFGE grouped 21 isolates into eight different clusters. Six of eight cluster I strains, all of four cluster II strains, and all of three cluster III strains were isolated in 2000, 2002, and 2003, respectively. Data concluded that P. aeruginosa carrying bla (VIM-2) with a class 1 integron was the only type among MBLs. A hospital outbreak by VIM-2 producers occurred annually, which could be at least a part of a recent high prevalence of imipenem resistance. PMID:16359195

  4. Clonal relatedness and conserved integron structures in epidemiologically unrelated Pseudomonas aeruginosa strains producing the VIM-1 metallo-{beta}-lactamase from different Italian hospitals.

    Science.gov (United States)

    Riccio, Maria Letizia; Pallecchi, Lucia; Docquier, Jean-Denis; Cresti, Stefania; Catania, Maria Rosaria; Pagani, Laura; Lagatolla, Cristina; Cornaglia, Giuseppe; Fontana, Roberta; Rossolini, Gian Maria

    2005-01-01

    Three epidemiologically independent Pseudomonas aeruginosa isolates, representative of the first VIM-1 metallo-beta-lactamase producers detected at three different hospitals in northern Italy, were investigated to determine their genomic relatedness and to compare the structures of the genetic supports for the VIM-1 determinants. The three isolates, all of serotype O11, appeared to be clonally related according to the results of genotyping by macrorestriction analysis of genomic DNA by pulsed-field gel electrophoresis and random amplification of polymorphic DNA. Investigation of the genetic support for the bla(VIM-1) determinant revealed that it was carried on identical or almost identical integrons (named In70.2 and In70.3) located within a conserved genomic context. The integrons were structurally related to In70 and In110, two plasmid-borne bla(VIM-1)-containing integrons from Achromobacter xylosoxidans and Pseudomonas putida isolates, respectively, from the same geographic area (northern Italy) and were found to be inserted close to the res site of a Tn5051-like transposon, different from any of those described previously, that was apparently carried on the bacterial chromosome. The present findings suggest that the three VIM-1-producing isolates are members of the same clonal complex which have been spreading in hospitals in northern Italy since the late 1990s and point to a common ancestry of their bla(VIM-1)-containing integrons. PMID:15616282

  5. Synthesis of silver nanoparticles using the Streptomyces coelicolor klmp33 pigment: An antimicrobial agent against extended-spectrum beta-lactamase (ESBL) producing Escherichia coli

    International Nuclear Information System (INIS)

    The increasing emergence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli) occurred mainly due to continuous persistent exposure to antibiotics causing high morbidity and mortality so studies in controlling this infection are required. In the present investigation, we developed a synthesis for silver nanoparticles employing a pigment produced by Streptomyces coelicolor klmp33, and assessed the antimicrobial activity of these nanoparticles against ESBL producing E. coli. The ESBL producing E. coli were isolated from urine samples collected from the Gulbarga region in India. As can been seen from our studies, the silver nanoparticles having irregular shapes and size of 28–50 nm showed remarkable antimicrobial activity and moreover the synthesis time is just 20 min and thus the same can be used for formulating pharmaceutical remedies. - Highlights: • Silver nanoparticle synthesis by photo-irradiation method in just 20 min • Isolation of ESBL producing E. coli from urine samples from the Gulbarga region. • Antimicrobial activity of silver nanoparticles against ESBL producing E. coli • The minimum inhibitory concentration of silver nanoparticles against ESBL producing E. coli was 40 μL

  6. Antimicrobial activity of antibiotics in combination with natural flavonoids against clinical extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Lin, Rong-Dih; Chin, Yi-Ping; Lee, Mei-Hsien

    2005-07-01

    Extended-spectrum beta-lactamases (ESBLs) are plasmid-mediated class A enzymes commonly found in the family Enterobacteriaceae, mainly in Klebsiella pneumoniae. Flavonoids have also been reported to possess antimicrobial activity. In this study, the in vitro activities of 18 antibiotics and 12 flavonoids against 20 ESBL-producing K. pneumoniae isolates were evaluated. All of these isolates were susceptible to imipenem and cefmetazole, but were resistant to ampicillin, ampicillin/sulbactam, aztreonam, cefazolin, cefoperazone, cefotaxime, ceftazidime, ceftriaxone, cefuroxime, piperacillin and ticarcillin. Susceptibilities to amikacin, amoxicillin/clavulanate, cefoxitin, ciprofloxacin and gentamicin were variable. Myricetin, a flavonol, inhibited ESBL-producing K. pneumoniae isolates at a high minimum inhibitory concentration (MIC) (MIC(90) value 256 mg/mL), but exhibited significant synergic activity against ESBL-producing K. pneumoniae in separate combination with amoxicillin/clavulanate, ampicillin/sulbactam and cefoxitin. Because of the low-toxic nature of flavonoids, the combination of antibiotics and flavonoids is a potential new strategy for developing therapies for infections caused by ESBL-producing bacteria in the future. PMID:16161024

  7. Characterization of extended-spectrum beta-lactamase (ESBL)-carrying plasmids and clones of Enterobacteriaceae causing cattle mastitis in France.

    Science.gov (United States)

    Dahmen, Safia; Métayer, Véronique; Gay, Emilie; Madec, Jean-Yves; Haenni, Marisa

    2013-03-23

    Extended-spectrum beta-lactamases (ESBLs) have become widespread enzymes in food-producing and companion animals worldwide. However, in cattle mastitis, a major cause of economic loss in the dairy industry, ESBL-producers were rarely described. In this study, from a collection of 1427 Escherichia coli and Klebsiella pneumoniae isolates causing clinical mastitis in France, we report 0.4% (6/1427) of the isolates carrying an ESBL gene. These six isolates were genetically unrelated and recovered over a 3-year period of time. The bla(CTX-M-14) gene was found in 4/6 isolates, and was predominantly located on F2:A-:B- IncFII plasmids. The bla(CTX-M-1) IncI1/ST3, which is widespread in various animal species in France, was found as well. Interestingly, among the five E. coli isolates, the ST23 and ST58 clones were found twice, together with the ST10 clone, all of which were previously found as ESBL-carriers in humans. Despite the very limited number of ESBL-producers recovered, this study shows a surprisingly low molecular diversity of the strains causing mastitis in France with respect to ESBL genes, plasmids and clones. Further work is needed to understand the major driving forces of the ESBL epidemiology in animals, including for different infections within the same animal species. PMID:23127568

  8. Antimicrobial resistance in invasive non-typhoid Salmonella from the Democratic Republic of the Congo: emergence of decreased fluoroquinolone susceptibility and extended-spectrum beta lactamases.

    Directory of Open Access Journals (Sweden)

    Octavie Lunguya

    Full Text Available BACKGROUND: Co-resistance against the first-line antibiotics ampicillin, chloramphenicol and trimethoprim/sulphamethoxazole or multidrug resistance (MDR is common in non typhoid Salmonella (NTS. Use of alternative antibiotics, such as fluoroquinolones or third generation cephalosporins is threatened by increasing resistance, but remains poorly documented in Central-Africa. METHODOLOGY/PRINCIPAL FINDINGS: As part of a microbiological surveillance study in DR Congo, blood cultures were collected between 2007 and 2011. Isolated NTS were assessed for serotype and antimicrobial resistance including decreased ciprofloxacin susceptibility and extended-spectrum beta-lactamase (ESBL production. In total, 233 NTS isolates (representing 23.6% of clinically significant organisms were collected, mainly consisting of Salmonella Typhimurium (79% and Salmonella Enteritidis (18%. The majority of NTS were isolated in the rainy season, and recovered from children ≤2 years old. MDR, decreased ciprofloxacin susceptibility, azithromycin and cefotaxime resistance were 80.7%, 4.3%, 3.0% and 2.1% respectively. ESBL production was noted in three (1.3% isolates. Decreased ciprofloxacin susceptibility was associated with mutations in codon 87 of the gyrA gene, while ESBLs all belonged to the SHV-2a type. CONCLUSIONS/SIGNIFICANCE: Presence of almost full MDR among NTS isolates from blood cultures in Central Africa was confirmed. Resistance to fluoroquinolones, azithromycin and third generation cephalosporins is still low, but emerging. Increased microbiological surveillance in DR Congo is crucial for adapted antibiotic therapy and the development of treatment guidelines.

  9. Spread of extended-spectrum beta-lactamase producing Escherichia coli isolates in Swedish broilers mediated by an incI plasmid carrying blaCTX-M-1

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    Börjesson Stefan

    2013-01-01

    Full Text Available Abstract Background The already high and increasing occurrence of extended-spectrum beta-lactamases (ESBL producing Escherichia coli in European broiler populations is of concern due to the fact that third and fourth generation cephalosporins are deemed critically important in human medicine. In Sweden 34% of the broilers carry ESBL/pAmpC producing E. coli in their gut, despite the absence of a known selection pressure such as antimicrobial usages. The aim of the current study was to characterise a selection of E. coli strains carrying the blaCTX-M-1, to determine if the spread was due to a specific clone. Findings Ten isolates carrying blaCTX-M-1 from Swedish broilers belonged to eight different multi-locus sequence types with three isolates belonging to ST155. The ST155 isolates were identical as assessed by PFGE. The blaCTX-M-1 was in all isolates carried on a plasmid of replicon type incI, which also transferred resistance to tetracycline and sulfamethoxazole. Conclusion The occurrence of ESBL-producing E. coli in the Swedish broilers is not due to the emergence of a single clone, but rather the spread of a specific incI plasmid carrying blaCTX-M-1.

  10. The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing

    Energy Technology Data Exchange (ETDEWEB)

    Chen Hao [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China); Shu Weiqun, E-mail: west2003@sohu.co [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China); Chang Xiaosong; Chen Jian; Guo Yebin; Tan Yao [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China)

    2010-07-15

    The spreading of extended-spectrum {beta}-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and bla{sub TEM+CTx-M} was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes. - Yangtze River is heavily polluted by ESBL-producing TC bacteria and Class 1 integrons play an important role in multi-drug resistance.

  11. Silver nanoparticle production by Rhizopus stolonifer and its antibacterial activity against extended spectrum {beta}-lactamase producing (ESBL) strains of Enterobacteriaceae

    Energy Technology Data Exchange (ETDEWEB)

    Banu, Afreen [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India); Rathod, Vandana, E-mail: drvandanarathod@rediffmail.com [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India); Ranganath, E. [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India)

    2011-09-15

    Highlights: {yields} Silver nanoparticle production by using Rhizopus stolonifer. {yields} Antibacterial activity of silver nanoparticles against extended spectrum {beta}-lactamase producing (ESBL) strains of Enterobacteriaceae. {yields} Synergistic effect of antibiotics with silver nanoparticles towards ESBL-strains. {yields} Characterization of silver nanoparticles made by UV-vis spectra, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transformed infrared (FTIR) spectroscopy, atomic force microscopy (AFM). -- Abstract: This report focuses on the synthesis of silver nanoparticles using the fungus, Rhizopus stolonifer and its antimicrobial activity. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Characterization of newly synthesized silver nanoparticles was made by UV-visible absorption spectroscopy, scanning electron microscope (SEM), transmission electron microscope (TEM), Fourier transform infrared (FTIR) spectroscopy and atomic force microscope (AFM). TEM micrograph revealed the formation of spherical nanoparticles with size ranging between 3 and 20 nm. The biosynthesized silver nanoparticles (AgNPs) showed excellent antibacterial activity against ESBL-strains which includes E. coli, Proteus. sp. and Klebsiella sp.

  12. Presence of Clostridium difficile and antibiotic and beta-lactamase activities in feces of volunteers treated with oral cefixime, oral cefpodoxime proxetil, or placebo.

    Science.gov (United States)

    Chachaty, E; Depitre, C; Mario, N; Bourneix, C; Saulnier, P; Corthier, G; Andremont, A

    1992-09-01

    Three groups of six healthy adult volunteers were randomly assigned to a treatment with 400 mg of oral cefpodoxime proxetil, oral cefixime, or placebo per day for 10 days. Informed consent was obtained from all volunteers. Clostridium difficile was not detected in the feces of any subject before treatment or at any time in the subjects in the placebo group. C. difficile was, however, detected in all subjects treated with cefpodoxime proxetil and in five of six treated with cefixime. Genomic DNA restriction patterns showed that the strains of C. difficile differed from one volunteer to another. Two subjects both shed different strains at different times during the 25-day surveillance period. All isolates were resistant to cefixime and cefpodoxime (MIC for 90% of strains, 256 and 512 mg/liter, respectively). Antibiotic activity was found in the feces of one volunteer treated with cefpodoxime proxetil and of four volunteers treated with cefixime. It was inversely correlated with the presence of fecal beta-lactamase activity. Intestinal side effects were limited to modifications of stool consistency, which occurred in only 3 of the 12 treated volunteers and did not lead to cessation of treatment. These modifications were significantly associated with the presence of fecal antibiotic activity (P less than 0.05) but not with the shedding of toxigenic or nontoxigenic strains of C. difficile or with the presence of toxin A in feces, which was detected only in one perfectly healthy treated volunteer. PMID:1416894

  13. Epidemiology and molecular characterization of extended-spectrum beta-lactamase-producing Enterobacter spp., Pantoea agglomerans, and Serratia marcescens isolates from a Bulgarian hospital.

    Science.gov (United States)

    Markovska, Rumyana Donkova; Stoeva, Temenuga Jekova; Bojkova, Kalina Dineva; Mitov, Ivan Gergov

    2014-04-01

    Forty-two extended-spectrum beta-lactamase (ESBL)-producing isolates of Enterobacter aerogenes, Enterobacter cloacae, Pantoea agglomerans, and Serratia marcescens, collected consecutively during the period January-November 2011 from the University Hospital in Varna, Bulgaria, were studied to characterize their ESBLs by isoelectric focusing, group-specific PCR, and sequencing. The epidemiological relationship was evaluated by random amplified polymorphic DNA analysis (RAPD). Transferability of ESBL genes was determined by conjugation experiments. Plasmid analysis was done by replicon typing and PstI fingerprinting. The overall rate of ESBL production was 20%. The most widespread enzyme was CTX-M-3, found in 64%. It was dominant in E. aerogenes (100%) and S. marcescens (83%). SHV-12, CTX-M-3, and CTX-M-15 were found among E. cloacae isolates in 50%, 35%, and 45%, respectively. Three main CTX-M-3-producing epidemic clones of E. aerogenes and S. marcescens have been detected. Among E. cloacae isolates, six different RAPD profiles were discerned. The plasmids harboring blaCTX-M-3 belonged to IncL/M type and demonstrated similar PstI fingerprinting profiles. IncFII plasmids were detected in two CTX-M-15-producing E. cloacae isolates. Our results demonstrate wide intrahospital dissemination of clonal E. aerogenes and S. marcescens isolates, carrying IncL/M conjugative plasmids. PMID:24171449

  14. Suspected nosocomial infections with multi-drug resistant E. coli, including extended-spectrum beta-lactamase (ESBL)-producing strains, in an equine clinic.

    Science.gov (United States)

    Walther, Birgit; Lübke-Becker, Antina; Stamm, Ivonne; Gehlen, Heidrun; Barton, Ann Kristin; Janssen, Traute; Wieler, Lothar H; Guenther, Sebastian

    2014-01-01

    Enterobacteriaceae such as Escherichia coli are common commensals as well as opportunistic and obligate pathogens. They cause a broad spectrum of infectious diseases in various hosts, including hospital-associated infections. In recent years, the rise of extended spectrum beta-lactamase (ESBL)-producing E. coli in companion animals (dogs, cats and horses) has been striking. However, reports on nosocomial infections are mostly anecdotic. Here we report on the suspected nosocomial spread of both ESBL-producing and non-ESBL-producing multi-drug resistant E. coli isolates in three equine patients within an equine clinic. Unlike easy-to-clean hospitalization opportunities available for small animal settings like boxes and cages made of ceramic floor tiles or stainless steel, clinical settings for horses are challenging environments for infection control programs due to unavoidable extraneous material including at least hay and materials used for horse bedding. The development of practice-orientated recommendations is needed to improve the possibilities for infection control to prevent nosocomial infections with multi-drug resistant and other transmissible pathogens in equine clinical settings. PMID:25872251

  15. Synthesis of silver nanoparticles using the Streptomyces coelicolor klmp33 pigment: An antimicrobial agent against extended-spectrum beta-lactamase (ESBL) producing Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Manikprabhu, Deene; Lingappa, K., E-mail: lingappak123@gmail.com

    2014-12-01

    The increasing emergence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli) occurred mainly due to continuous persistent exposure to antibiotics causing high morbidity and mortality so studies in controlling this infection are required. In the present investigation, we developed a synthesis for silver nanoparticles employing a pigment produced by Streptomyces coelicolor klmp33, and assessed the antimicrobial activity of these nanoparticles against ESBL producing E. coli. The ESBL producing E. coli were isolated from urine samples collected from the Gulbarga region in India. As can been seen from our studies, the silver nanoparticles having irregular shapes and size of 28–50 nm showed remarkable antimicrobial activity and moreover the synthesis time is just 20 min and thus the same can be used for formulating pharmaceutical remedies. - Highlights: • Silver nanoparticle synthesis by photo-irradiation method in just 20 min • Isolation of ESBL producing E. coli from urine samples from the Gulbarga region. • Antimicrobial activity of silver nanoparticles against ESBL producing E. coli • The minimum inhibitory concentration of silver nanoparticles against ESBL producing E. coli was 40 μL.

  16. Metallo-beta-lactamases in clinical Pseudomonas isolates in Taiwan and identification of VIM-3, a novel variant of the VIM-2 enzyme.

    Science.gov (United States)

    Yan, J J; Hsueh, P R; Ko, W C; Luh, K T; Tsai, S H; Wu, H M; Wu, J J

    2001-08-01

    A total of 209 clinical isolates of Pseudomonas (193 Pseudomonas aeruginosa, 10 P. putida, 4 P. stutzeri, and 2 P. fluorescens isolates) with reduced susceptibilities to imipenem and/or ceftazidime were subjected to PCR assays with primers specific for bla(IMP-1), bla(IMP-2), bla(VIM-1), and bla(VIM-2) and sequence analysis to identify the metallo-beta-lactamases (MBLs) prevalent among these organisms in Taiwan; and 21 isolates gave positive results. Five isolates including two P. putida and three P. stutzeri isolates were found to carry bla(IMP-1), and six isolates including five P. putida and one P. stutzeri isolates harbored bla(VIM-2). The remaining 10 isolates were P. aeruginosa, and all were found to carry a novel variant of bla(VIM-2), designated bla(VIM-3). There are only two nucleotide differences between bla(VIM-2) and bla(VIM-3), leading to two amino acid alterations. Our findings indicate that VIM-2 and its variant have become the most prevalent metalloenzymes in Pseudomonas in Taiwan. Southern hybridization with the bla(VIM-2)-, bla(VIM-3)-, and bla(IMP-1 )-specific probes revealed that only two VIM-2-producing P. putida isolates appeared to carry the MBL gene on plasmids. Pulsed-field gel electrophoresis showed that six VIM-3-producing P. aeruginosa isolates and two IMP-1-producing P. stutzeri isolates were genetically related, suggesting that the spread of these MBL genes in Taiwan could be due to clonal dissemination as well as genetic exchange between different clones. PMID:11451678

  17. PCR typing of genetic determinants for metallo-beta-lactamases and integrases carried by gram-negative bacteria isolated in Japan, with focus on the class 3 integron.

    Science.gov (United States)

    Shibata, Naohiro; Doi, Yohei; Yamane, Kunikazu; Yagi, Tetsuya; Kurokawa, Hiroshi; Shibayama, Keigo; Kato, Haru; Kai, Kumiko; Arakawa, Yoshichika

    2003-12-01

    From January 2001 to December 2002, 587 strains of gram-negative bacterial isolates demonstrating resistance to ceftazidime and a combination of sulbactam and cefoperazone were subjected to a disk diffusion screening test using sodium mercaptoacetic acid; 431 strains (73.4%) appeared to produce metallo-beta-lactamase (MBL). Of these 431 strains, 357 were found by PCR to carry genes for IMP-1 type MBL (bla(IMP-1)), while only 7 and 67 strains carried the IMP-2 gene (bla(IMP-2)) and the VIM-2 gene (bla(VIM-2)), respectively. Neither VIM-1 nor SPM-1 type MBL genes were found among the strains tested. Of 431 strains, 427 carried the intI1 gene, and 4 strains carrying both the intI1 and intI3 genes were reidentified as Pseudomonas putida harboring bla(IMP-1). Of these four P. putida strains, three strains and one strain, respectively, were separately isolated from two hospitals located in the same prefecture, and the three strains showed very similar pulsed-field gel electrophoresis patterns. Of 357 bla(IMP-1) carriers, 116, 53, 51, 47, and 30 strains were identified as Pseudomonas aeruginosa, Alcaligenes xylosoxidans, P. putida/fluorescens, Serratia marcescens, and Acinetobacter baumannii, respectively. Four strains carrying bla(IMP-2) were reidentified as P. putida. Sixty-three P. aeruginosa strains and four P. putida strains carried bla(VIM-2). Of 427 intI1-positive strains, 180, 53, 51, 47, and 35 were identified as P. aeruginosa, A. xylosoxidans, P. putida/fluorescens, S. marcescens, and A. baumannii, respectively. In the present study, it was confirmed that strains carrying bla(IMP-1) with a class 1 integron are the most prevalent type in Japan, although several intI3 carriers have also been identified sporadically in this country. PMID:14662918

  18. Characterization of the new metallo-beta-lactamase VIM-13 and its integron-borne gene from a Pseudomonas aeruginosa clinical isolate in Spain.

    Science.gov (United States)

    Juan, Carlos; Beceiro, Alejandro; Gutiérrez, Olivia; Albertí, Sebastián; Garau, Margalida; Pérez, José L; Bou, Germán; Oliver, Antonio

    2008-10-01

    During a survey conducted to evaluate the incidence of class B carbapenemase (metallo-beta-lactamase [MBL])-producing Pseudomonas aeruginosa strains from hospitals in Majorca, Spain, five clinical isolates showed a positive Etest MBL screening test result. In one of them, strain PA-SL2, the presence of a new bla(VIM) derivative (bla(VIM-13)) was detected by PCR amplification with bla(VIM-1)-specific primers followed by sequencing. The bla(VIM-13)-producing isolate showed resistance to all beta-lactams (except aztreonam), gentamicin, tobramycin, and ciprofloxacin. VIM-13 exhibited 93% and 88% amino acid sequence identities with VIM-1 and VIM-2, respectively. bla(VIM-13) was cloned in parallel with bla(VIM-1), and the resistance profile conferred was analyzed both in Escherichia coli and in P. aeruginosa backgrounds. Compared to VIM-1, VIM-13 conferred slightly higher levels of resistance to piperacillin and lower levels of resistance to ceftazidime and cefepime. VIM-13 and VIM-1 were purified in parallel as well, and their kinetic parameters were compared. The k(cat)/K(m) ratios for the antibiotics mentioned above were in good agreement with the MIC data. Furthermore, EDTA inhibited the activity of VIM-13 approximately 25 times less than it inhibited the activity of VIM-1. VIM-13 was harbored in a class 1 integron, along with a new variant (Ala108Thr) of the aminoglycoside-modifying enzyme encoding gene aacA4, which confers resistance to gentamicin and tobramycin. Finally, the VIM-13 integron was apparently located in the chromosome, since transformation and conjugation experiments consistently yielded negative results and the bla(VIM-13) probe hybridized only with the genomic DNA. PMID:18644957

  19. Extensive Within-Host Diversity in Fecally Carried Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli Isolates: Implications for Transmission Analyses.

    Science.gov (United States)

    Stoesser, N; Sheppard, A E; Moore, C E; Golubchik, T; Parry, C M; Nget, P; Saroeun, M; Day, N P J; Giess, A; Johnson, J R; Peto, T E A; Crook, D W; Walker, A S

    2015-07-01

    Studies of the transmission epidemiology of antimicrobial-resistant Escherichia coli, such as strains harboring extended-spectrum beta-lactamase (ESBL) genes, frequently use selective culture of rectal surveillance swabs to identify isolates for molecular epidemiological investigation. Typically, only single colonies are evaluated, which risks underestimating species diversity and transmission events. We sequenced the genomes of 16 E. coli colonies from each of eight fecal samples (n = 127 genomes; one failure), taken from different individuals in Cambodia, a region of high ESBL-producing E. coli prevalence. Sequence data were used to characterize both the core chromosomal diversity of E. coli isolates and their resistance/virulence gene content as a proxy measure of accessory genome diversity. The 127 E. coli genomes represented 31 distinct sequence types (STs). Seven (88%) of eight subjects carried ESBL-positive isolates, all containing blaCTX-M variants. Diversity was substantial, with a median of four STs/individual (range, 1 to 10) and wide genetic divergence at the nucleotide level within some STs. In 2/8 (25%) individuals, the same blaCTX-M variant occurred in different clones, and/or different blaCTX-M variants occurred in the same clone. Patterns of other resistance genes and common virulence factors, representing differences in the accessory genome, were also diverse within and between clones. The substantial diversity among intestinally carried ESBL-positive E. coli bacteria suggests that fecal surveillance, particularly if based on single-colony subcultures, will likely underestimate transmission events, especially in high-prevalence settings. PMID:25903575

  20. Empiric Piperacillin-Tazobactam versus Carbapenems in the Treatment of Bacteraemia Due to Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae

    Science.gov (United States)

    Harris, Patrick N. A.; De, Partha P.; Chow, Angela; Tambyah, Paul A.; Lye, David C.

    2016-01-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae are a common cause of bacteraemia in endemic countries and may be associated with high mortality; carbapenems are considered the drug of choice. Limited data suggest piperacillin-tazobactam could be equally effective. We aimed to compare 30-day mortality of patients treated empirically with piperacillin-tazobactam versus a carbapenem in a multi-centre retrospective cohort study in Singapore. Only patients with active empiric monotherapy with piperacillin-tazobactam or a carbapenem were included. A propensity score for empiric carbapenem therapy was derived and an adjusted multivariate analysis of mortality was conducted. A total of 394 patients had ESBL-Escherichia.coli and ESBL-Klebsiella pneumoniae bacteraemia of which 23.1% were community acquired cases. One hundred and fifty-one received initial active monotherapy comprising piperacillin-tazobactam (n = 94) or a carbapenem (n = 57). Patients who received carbapenems were less likely to have health-care associated risk factors and have an unknown source of bacteraemia, but were more likely to have a urinary source. Thirty-day mortality was comparable between those who received empiric piperacillin-tazobactam and a carbapenem (29 [30.9%] vs. 17 [29.8%]), P = 0.89). Those who received empiric piperacillin-tazobactam had a lower 30-day acquisition of multi-drug resistant and fungal infections (7 [7.4%] vs. 14 [24.6%]), P<0.01). After adjusting for confounders, use of empiric piperacillin-tazobactam was not associated with increased 30-day mortality (OR 1.00, 95% CI; 0.45–2.17). Empiric piperacillin-tazobactam was not associated with increased 30-day mortality and may result in fewer multi-drug resistant and fungal infections when compared with a carbapenem. PMID:27104951

  1. Detection of extended spectrum beta-lactamases and resistance in members of the Enterobacteriaceae family isolated from healthy sheep and dogs in Umuarama, Paraná, Brazil

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    Patricia Alves de Oliveira

    2016-04-01

    Full Text Available Bacterial resistance is a primary public health concern worldwide. Within this context, pets and breeding animals act as reservoirs for multidrug-resistant bacteria (MR, such as those producing extended spectrum beta-lactamases (ESBL and those presenting plasmid-mediated quinolone resistance (PMQR. The aim of this study was to detect the presence of ESBL and PMQR in members of the Enterobacteriaceae family, isolated from healthy sheep and dogs from non-intense farming rural properties in the Umuarama region of Paraná, Brazil. A total of 81 oral and rectal swabs from dogs and sheep from 11 small rural properties were analyzed. These swabs were inoculated in tubes containing brain heart infusion broth (BHI, and the resulting cultures were inoculated on MacConkey agar (MAC supplemented with 10 ?g/mL cefotaxime for the selection of ESBL producers. The cells were also plated on MAC supplemented with 50 ?g/mL nalidixic acid for selecting quinolone-resistant enterobacteria. The bacterial isolates were subjected to biochemical identification tests, antibiograms, double-disk synergic tests, and polymerase chain reaction analysis for resistance-inducing genes (blaESBL, qnr, and genes encoding efflux pump and acetylases. Four (5.00% bacterial isolates (3 Escherichia coli and 1 Morganella morganii resistant to cephalosporins and/or quinolones were identified; of these, three (75% isolates were from sheep and one (25% from a dog. These findings indicate the presence of MR bacteria in the normal microbiota of the animals studied. Animals colonized with such bacteria can contribute to the dissemination of antimicrobial resistance to other animals, environment, and/or human beings and can harbor endogenous infections in unfavorable conditions, which have poor prognosis due to the limited therapeutic options.

  2. Extended-spectrum beta-lactamase-producing bacteria are not detected in supragingival plaque samples from human fecal carriers of ESBL-producing Enterobacteriaceae

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    Arne Søraas

    2014-08-01

    Full Text Available Background: The prevalence of infections caused by Cefotaximase-Munich (CTX-M-type extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E has rapidly increased during the past 15 years. Enterobacteriaceae are commonly found in the gastrointestinal tract and long-term intestinal carriage is considered important for the spread of ESBL and as a source of clinical infections. Oral biofilm such as supragingival plaque is known to contain numerous antibiotic resistance determinants and may also represent a poorly investigated site for ESBL carriage and further spread. Objective: To investigate possible carriage of ESBL-producing bacteria in supragingival plaque of known fecal carriers of these bacteria. Design: We screened for the presence of aerobic and anaerobic ESBL-producing bacteria and blaCTX-M in supragingival plaque samples from healthy human adults with culture-verified fecal carriage of CTX-M-producing Escherichia coli. The presence or absence of Enterobacteriaceae and ESBL-producing bacteria in plaque samples was evaluated using culture-based methods and consensus CTX-M PCR. Results: Oral samples were obtained from 17 participants with known previous carriage of ESBL-producing E. coli. No ESBL-producing bacteria or ESBL genes were detected using culture-based and molecular methods. One colony of Rahnella aquatilis harboring the class A ESBL gene bla RAHN-1/2 was identified in an oral sample from one of the participants. Conclusion: This pilot study supports the notion that the presence of CTX-M-producing bacteria is uncommon in oral plaque of healthy human adult fecal carriers. Due to the limited number of persons tested, a low prevalence of oral ESBL-carriage in healthy adults or carriage in selected groups of patients cannot be excluded. To our knowledge, this is the first description of an R. aquatilis with the RAHN-1/2 gene in the oral cavity.

  3. Prevalence of CTX-M extended-spectrum beta-lactamases and sequence type 131 in Korean blood, urine, and rectal Escherichia coli isolates.

    Science.gov (United States)

    Graham, Sarah E; Zhang, Lixin; Ali, Ihsan; Cho, Yong Kyun; Ismail, Miriam D; Carlson, Heather A; Foxman, Betsy

    2016-07-01

    A high proportion of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli are of the ST131 lineage, but there are few estimates of ST131 prevalence among ESBL-negative E. coli. Without this information, it is difficult to evaluate the contribution of the ST131 lineage to the emergence and spread of ESBL E. coli. A total of 1658 E. coli isolates were collected at Gachon University Gil Medical Center in Korea from 2006 to 2008. The antibiotic resistance profile was determined for all isolates, and ESBL-positive isolates were screened for the presence of CTX-M-type ESBLs. All ESBL-positive (n=84) and a representative sample of ESBL-negative (n=100) isolates were screened for O25b-ST131 using a PCR-based assay. The isolates were further classified on the basis of fumC and fimH types, which allowed for a comparison of the two typing methods. 5.7% of isolates were ESBL-positive, 87% of which contained CTX-M-type ESBLs. There was no significant difference in the prevalence of ST131 between ESBL-positive and -negative groups; 14% of ESBL-positive isolates and 9% of tested ESBL-negative isolates were ST131 by CH-typing. ST131-positive isolates harbored CTX-M-1-group ESBLs (including CTX-M-15) more frequently than other CTX-M types, and exhibited greater levels of antibiotic resistance than non-ST131 isolates. Furthermore, a number of isolates identified as O25b-ST131 by PCR corresponded to non-ST131 sequence types by CH-typing, emphasizing the need to consider the testing method when comparing reported prevalences of ST131. PMID:27101781

  4. Evaluation of Zataria MultiFlora Boiss and Carum copticum antibacterial activity on IMP-type metallo-beta-lactamase-producing Pseudomonas aeruginosa.

    Science.gov (United States)

    Fallah, F; Taherpour, A; Borhan, R S; Hashemi, A; Habibi, M; Sajadi Nia, R

    2013-12-31

    Carbapenem resistance due to acquired metallo-beta-lactamases (MBLs) is considered to be more serious than other resistance mechanisms. The aim of this study was to evaluate the antibacterial activity of Zataria multiflora Boiss and Carum copticum plants on IMP-producing P.aeruginosa strains. This experimental study was carried out on hospitalized burn patients during 2011 and 2012. Antibiotics and extracts susceptibility tests were performed by disc diffusion and broth microdilution methods. MBL detection was performed by Combination Disk Diffusion Test (CDDT). The bla(VIM) and bla(IMP) genes were detected by PCR and sequencing methods. Using Combination Disk Diffusion test method, it was found that among 83 imipenem resistant P.aeruginosa strains, 48 (57.9%) were MBL producers. PCR and sequencing methods proved that these isolates were positive for blaIMP-1 genes, whereas none were positive for bla(VIM) genes. The mortality rate of hospitalized patients with MBL-producing Pseudomonas infection was 4/48 (8.3%). It was shown that Zataria multiflora and Carum copticum extracts had a high antibacterial effect on regular and IMP-producing P. aeruginosa strains in 6.25 mg/ml concentration. The incidence of MBL-producing P. aeruginosa in burn patients is very high. In our study, all MBL-producing isolates carry the blaIMP-1 gene. Therefore, detection of MBL-producing isolates is of great importance in identifying drug resistance patterns in P. aeruginosa, and in prevention and control of infections. In this study, it was shown that extracts of Z. multiflora and C. copticum have high antibacterial effects on ß-lactamase producing P. aeruginosa strains. PMID:24799849

  5. First description of Escherichia coli producing CTX-M-15- extended spectrum beta lactamase (ESBL in out-patients from south eastern Nigeria

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    Iroha Ifeanyichukwu R

    2012-07-01

    Full Text Available Abstract We studied the presence of extended spectrum beta lactamases (ESBLs in 44 clinical isolates of Escherichia coli collected from out-patients in two university teaching hospitals in South-Eastern Nigeria. Species identification was performed by standard microbiology methods and re-confirmed by MALDI-TOF technology. Phenotypic characterization of ESBL enzymes was done by double disc synergy test and presence of ESBL genes was determined by specific PCR followed by sequencing. Transfer of plasmid DNA was carried out by transformation using E. coli DH5 as recipient strain. Phenotypic characterization identified all isolates to be ESBL positive. 77% of strains were from urine, 13.6% from vaginal swabs and 9.0% from wound swabs. 63.6% were from female patients, 68% were from outpatients and 95.5% from patients younger than 30 years. All ESBL producers were positive in a PCR for blaCTX-M-1 cluster, in exemplary strains blaCTX-M-15 was found by sequencing. In all strains ISEcp1 was found upstream and ORF477 downstream of blaCTX-M. PCR for blaTEM and blaOXA-1 was positive in 93.1% of strains, whereas blaSHV was not detected, aac(6′-Ib-cr was found in 97.7% of strains. RAPD analysis revealed seven different clonal groups named A through G with the majority of the strains (65.9% belonging to clone A. Transfer of an ESBL plasmid with co-resistance to gentamicin, kanamycin, tobramycin, doxycycline and trimethropim-sulfamethoxazole was successful in 19 (43.2% strains. This study showed a high rate of CTX-M-1 cluster - ESBLs in South-Eastern Nigeria and further confirms the worldwide spread of CTX-M ESBL in clinical isolates.

  6. Antimicrobial resistance among producers and non-producers of extended spectrum beta-lactamases in urinary isolates at a tertiary Hospital in Tanzania

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    Lyamuya Eligius F

    2010-12-01

    Full Text Available Abstract Background Published data on the existence and magnitude of extended spectrum beta-lactamase (ESBL production in urinary pathogens in local setting is limited. The aim of the present study was to determine the prevalence of antimicrobial resistance and ESBL production among Escherichia coli and Klebsiella spp from urine samples in a tertiary hospital. This was a cross sectional study conducted at Muhimbili National Hospital in Dar es Salaam, Tanzania. Findings A total of 270 E.coli and Klebsiella spp urinary pathogens from children and adults isolated from January to March 2010 were included in the study. E. coli and Klebsiella spp isolates were tested for antimicrobial susceptibility by the Clinical and Laboratory Standard Institute's disc diffusion method. These isolates were further screened for ESBL phenotype using cefotaxime and ceftazidime discs. Isolates with reduced sensitivity were confirmed using ESBL E-test strips. Of 270 isolates, 138 (51.1% were E. coli and 132 (48.9% were Klebsiella spp. ESBL was detected in 122 (45.2% of all the isolates. ESBL- producing E. coli strains were significantly more resistance to cotrimoxazole (90.7%, ciprofloxacin (46.3% and nalidixic acid (61.6% than strains that did not produce ESBL (p Klebsiella spp strains were significantly more resistance to cotrimoxazole (92.6%, ciprofloxacin (25.0%, nalidixic acid (66.2%, and gentamicin (38.2% than strains that did not produce ESBL (P P Conclusions High prevalence of ESBL-producing E. coli and Klebsiella spp strains was found among inpatients and children. Most of the ESBL- producing isolates were multi-drug resistant making available therapeutic choices limited. We recommend continued antibiotic surveillance as well comprehensive multi-center studies to address the emerging problem of ESBL-associated infections in order to preserve the continued usefulness of most antimicrobial drugs. Further more conducting molecular studies will help to evaluate the

  7. A trial with IgY chicken antibodies to eradicate faecal carriage of Klebsiella pneumoniae and Escherichia coli producing extended-spectrum beta-lactamases

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    Anna-Karin Jonsson

    2015-11-01

    Full Text Available Background: Extended-spectrum beta-lactamase (ESBL-producing Enterobacteriaceae is an emerging therapeutic challenge, especially in the treatment of urinary tract infections. Following an outbreak of CTX-M-15 Klebsiella pneumoniae in Uppsala, Sweden, an orphan drug trial on IgY chicken antibodies was undertaken in an attempt to eradicate faecal carriage of ESBL-producing K. pneumoniae and Escherichia coli. Methods: Hens were immunised with epitopes from freeze-dried, whole-cell bacteria (ESBL-producing K. pneumoniae and E. coli and recombinant proteins of two K. pneumoniae fimbriae subunits (fimH and mrkD. The egg yolks were processed according to good manufacturing practice and the product was stored at−20°C until used. Using an internal database from the outbreak and the regular laboratory database, faecal carriers were identified and recruited from May 2005 to December 2013. The participants were randomised in a placebo-controlled 1:1 manner. Results: From 749 eligible patients, 327 (44% had deceased, and only 91 (12% were recruited and signed the informed consent. In the initial screening performed using the polymerase chain reaction, 24 participants were ESBL positive and subsequently randomised and treated with either the study drug or a placebo. The study was powered for 124 participants. Because of a very high dropout rate, the study was prematurely terminated. From the outbreak cohort (n=247, only eight patients were screened, and only one was positive with the outbreak strain in faeces. Conclusions: The present study design, using IgY chicken antibodies for the eradication of ESBL-producing K. pneumonia and E. coli, was ineffective in reaching its goal due to high mortality and other factors resulting in a low inclusion rate. Spontaneous eradication of ESBL-producing bacteria was frequently observed in recruited participants, which is consistent with previous reports.

  8. The prevalence of extended-spectrum beta-lactamases and CTX-M-1 producing Escherichia coli in urine samples collected at Tabriz city Hospitals

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    Soltan Dallal MM

    2011-08-01

    Full Text Available "nBackground: Numerous use of Beta Lactame in treatment of bacterial infections resulted in increments of drug resistance of such bacteria. One of difficulties in treatment of hospital infections is Extended Spectrum Beta Lactamase (ESBL among isolated clinical strains of E.coli. Since some of ESBL strains shows double reaction in drug sensitivity test at in vitro and in vivo condition, therefore it makes difficulties in selection of right treatment. In the last years, CTX-M enzymes have become the most prevalent ESBLs in worldwide. The prevalence of ESBL types largely remains unknown in many parts of the Iran. This study was made to find the prevalence of ESBL-producing E.coli and molecular detection of CTX-M-1 in Tabriz."n "nMethods: In the present study, 400 urine samples collected between November 2009 and April 2010, from Tabriz Hospitals were studied. Out of the 400 samples, 188 E.coli isolates were detected by standard biochemical tests. Susceptibility to antimicrobial agents was tested to 10 antibiotics by the disk agar diffusion (DAD method. ESBL production was screened by phenotypic test that included both separate and combined disk agar diffusion techniques. The screened isolates were investigated by PCR assay to detect CTX-M-1 gene."n "nResults: From the total 188 E.coli isolates, 82 (43.6% were shown to produce ESBLs by phenotypic test. During the PCR method on the 82 isolates, 69 (84.1% were confirmed as CTX-M-1 producing group."n "nConclusion: The present study showed that CTX-M-producing isolates were increasing among E.coli strains and indicated the need for adequate susceptibility tests in laboratories for choosing the appropriate antibiotics for treatment.

  9. CTX-M-2 and a New CTX-M-39 Enzyme Are the Major Extended-Spectrum Beta-Lactamases in Multiple Escherichia coli Clones Isolated in Tel Aviv, Israel

    OpenAIRE

    Chmelnitsky, Inna; Carmeli, Yehuda; Leavitt, Azita; Schwaber, Mitchell J.; Navon-Venezia, Shiri

    2005-01-01

    The rate of occurrence of the extended-spectrum beta-lactamase (ESBL)-producing phenotype among Escherichia coli isolates in Tel Aviv is 12% (22). The aim of this study was to understand the molecular epidemiology of E. coli ESBL producers and to identify the ESBL genes carried by them. We studied 20 single-patient ESBL-producing E. coli clinical isolates. They comprised 11 distinct nonrelated pulsed-field gel electrophoresis (PFGE) genotypes: six isolates belonged to the same PFGE clone, fou...

  10. Characterization of extended-spectrum beta-lactamases and antimicrobial resistance of Klebsiella pneumoniae in intra-abdominal infection isolates in Latin America, 2008-2012. Results of the Study for Monitoring Antimicrobial Resistance Trends.

    Science.gov (United States)

    Kazmierczak, Krystyna M; Lob, Sibylle H; Hoban, Daryl J; Hackel, Meredith A; Badal, Robert E; Bouchillon, Samuel K

    2015-07-01

    The Study for Monitoring Antimicrobial Resistance Trends has monitored the in vitro activity of several recommended antimicrobials used in the management of intra-abdominal infections (IAIs) globally since 2002. In this report, we document the changing susceptibility patterns to recommended antimicrobials in Klebsiella pneumoniae isolates from patients with IAIs in 11 Latin American countries between 2008 and 2012 and describe the beta-lactamases encoded by phenotypically extended-spectrum beta-lactamase (ESBL)-positive and ertapenem-nonsusceptible isolates. Overall, the incidence of phenotypically ESBL-positive K. pneumoniae did not change significantly from 2008 (40.4%) to 2012 (41.2%) (P > 0.05). However, trend analysis documented an increase in isolates encoding K. pneumoniae carbapenemase (KPC) or both KPC and an ESBL. Decreasing susceptibility (P ceftriaxone, ertapenem, and imipenem among all K. pneumoniae, as well as for cefepime, cefotaxime, cefoxitin, ceftriaxone, ertapenem, and imipenem among ESBL-positive isolates, while susceptibility of ESBL-negative isolates to ampicillin-sulbactam actually increased (P < 0.05). PMID:25956930

  11. Development of a multiplex real-time PCR for the rapid detection of the predominant beta-lactamase genes CTX-M, SHV, TEM and CIT-type AmpCs in Enterobacteriaceae.

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    Nicole Roschanski

    Full Text Available Beta-lactamase resistant bacteria and especially ESBL producing Enterobacteriaceae are an increasing problem worldwide. For this reason a major interest in efficient and reliable methods for rapid screening of high sample numbers is recognizable. Therefore, a multiplex real-time PCR was developed to detect the predominant class A beta-lactamase genes blaCTX-M, blaSHV, blaTEM and CIT-type AmpCs in a one-step reaction. A set of 114 Enterobacteriaceae containing previously identified resistance gene subtypes and in addition 20 undefined animal and environmental isolates were used for the validation of this assay. To confirm the accessibility in variable settings, the real-time runs were performed analogous in two different laboratories using different real-time cyclers. The obtained results showed complete accordance between the real-time data and the predetermined genotypes. Even if sequence analyses are further necessary for a comprehensive characterization, this method was proofed to be reliable for rapid screening of high sample numbers and therefore could be an important tool for e. g. epidemiological purposes or support infection control measures.

  12. Epidemiology and Burden of Bloodstream Infections Caused by Extended-Spectrum Beta-Lactamase Producing Enterobacteriaceae in a Pediatric Hospital in Senegal.

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    Awa Ndir

    Full Text Available Severe bacterial infections are not considered as a leading cause of death in young children in sub-Saharan Africa. The worldwide emergence of extended-spectrum beta-lactamase producing Enterobacteriaceae (ESBL-E could change the paradigm, especially in neonates who are at high risk of developing healthcare-associated infections.To evaluate the epidemiology and the burden of ESBL-E bloodstream infections (BSI.A case-case-control study was conducted in patients admitted in a pediatric hospital during two consecutive years. Cases were patients with Enterobacteriaceae BSI and included ESBL-positive (cases 1 and ESBL-negative BSI (cases 2. Controls were patients with no BSI. Multivariate analysis using a stepwise logistic regression was performed to identify risk factors for ESBL acquisition and for fatal outcomes. A multistate model was used to estimate the excess length of hospital stay (LOS attributable to ESBL production while accounting for time of infection. Cox proportional hazards models were performed to assess the independent effect of ESBL-positive and negative BSI on LOS.The incidence rate of ESBL-E BSI was of 1.52 cases/1000 patient-days (95% CI: 1.2-5.6 cases per 1000 patient-days. Multivariate analysis showed that independent risk factors for ESBL-BSI acquisition were related to underlying comorbidities (sickle cell disease OR = 3.1 (95%CI: 2.3-4.9, malnutrition OR = 2.0 (95%CI: 1.7-2.6 and invasive procedures (mechanical ventilation OR = 3.5 (95%CI: 2.7-5.3. Neonates were also identified to be at risk for ESBL-E BSI. Inadequate initial antibiotic therapy was more frequent in ESBL-positive BSI than ESBL-negative BSI (94.2% versus 5.7%, p<0.0001. ESBL-positive BSI was associated with higher case-fatality rate than ESBL-negative BSI (54.8% versus 15.4%, p<0.001. Multistate modelling indicated an excess LOS attributable to ESBL production of 4.3 days. The adjusted end-of-LOS hazard ratio for ESBL-positive BSI was 0.07 (95%CI, 0

  13. High Prevalence of Escherichia coli-Producing CTX-M-15 Extended-Spectrum Beta-Lactamases in Poultry and Human Clinical Isolates in Romania.

    Science.gov (United States)

    Maciuca, Iuliana E; Williams, Nicola J; Tuchilus, Cristina; Dorneanu, Olivia; Guguianu, Eleonora; Carp-Carare, Catalin; Rimbu, Cristina; Timofte, Dorina

    2015-12-01

    Use of antibiotics in food animals may contribute to development and spread of resistant organisms, particularly so in some countries. The aim of this study was two-fold; first, to establish the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in chicken production in a region within Romania. Second, to study the relatedness of ESBL-producing E. coli isolates recovered from broilers, abattoir workers where the chickens were slaughtered and from the human clinical specimens from two regional hospitals. The results indicated a very high (69%) rate of carriage of ESBL and AmpC-producing E. coli in chickens with 36% CTX-M producers. Sequencing showed that chickens in Romania have the highest worldwide prevalence (53%) of blaCTX-M-15 reported in poultry E. coli isolates. The majority (53%) of the extended-spectrum cephalosporin-resistant E. coli carried plasmid-mediated blaampC genes, mostly blaCMY-2 type, one of the highest prevalences reported in Europe. The predominant CTX-M type found in the human clinical E. coli isolates was blaCTX-M-15 and most isolates coharbored blaOXA-1, blaTEM, and aac(6')-ib-cr. The majority (60%) of the human clinical isolates belonged to the pandemic virulent clone B2-ST131. The clonal relationship between broiler and the human CTX-M-producing E. coli isolates was assessed by macrorestriction pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST), which indicated strain diversity with no common STs found between human and poultry isolates. Moreover, IncI1 was the most prevalent replicon found in broiler ESBL-producing E. coli isolates and also in transconjugants, indicating that plasmids and not clonal spread may play a role in the transfer of blaCTX-M genes. This study identifies a high prevalence of ESBL-producing E. coli from broiler chickens in Romania with a high occurrence incidence of blaCTX-M-15, which reflects the main ESBL type found in human E. coli infections in this

  14. Identification of Clonal Clusters of Klebsiella pneumoniae Isolates from Chennai by Extended Spectrum Beta Lactamase Genotyping and Antibiotic Resistance Phenotyping Analysis

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    C. Kamatchi

    2009-01-01

    Full Text Available Problem statement: An increased resistance to antibiotics has been reported in Klebsiella pneumoniae, an opportunistic gram negative pathogen belonging to Entrobacteraceae due to the evolution and spread of plasmid encoded extended spectrum beta lactamases and other genes conferring cross-resistance to other antibiotics. This is of concern due to the increasing cost of antibiotic treatment and the spread of multidrug resistance to more pathogenic microorganisms. This study was undertaken to analyze the extent of the problem, to identify the most prevalent MDR isolates of Klebsiella pneumoniae in Chennai and to identify new plant based drugs. Approach: About 188 clinical isolates of Klebsiella pneumoniae were collected from Chennai during the period Nov. 2007-Oct. 2008 and their resistance to different groups of antibiotics were analyzed. These isolates were further characterized by molecular studies to identify the ESBL genes conferring the resistance phenotype. Plant extracts were tested against the MDR isolates to identify new treatment methods. Results: Our results showed that the combination therapy of clavulanic acid with cephalosporins and fluroroquinolones-norfloxacin and ciprofloxacin were the most effective antibiotics for treatment of Klebsiella pneumoniae infections. However resistance to clavulanic acid was increasing among the isolates (17%. All the isolates harbored a plasmid containing one or more of the genes for ESBL resistance. Plasmid isolation from the Isolates followed by ESBL PCR genotyping showed that CTX-M was present in 75 % isolates and TEM in 73% isolates either alone or in combination with the other ESBL genes. SHV ESBL type was present only in 42% of isolates. We identified 4 presumptive clonal spreads which were likely to be prevalent in India by clustering based on ESBL genotypic and antibiotic resistance. The isolates with both SHV and CTX were correlated with the

  15. Antibacterial activities of multi drug resistant Myroides odoratimimus bacteria isolated from adult flesh flies (Diptera: Sarcophagidae are independent of metallo beta-lactamase gene Atividades antibacterianas de Myroides odoratimimus isolada de moscas varejeiras adultas (Diptera: Sarcophagidae são independentes do gene metalo beta lactamase

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    M.S. Dharne

    2008-06-01

    avaliação de parâmetros nutricionais pelo sistema BIOLOG GN, ao sequenciamento genético 16S rRNA, à sensibilidade a vários antimicrobianos pelo método de difusão de discos e à detecção dos genes de metalo beta lactamases (TUS/MUS. Os efeitos antagonistas foram testados contra bactérias Gram negativas e Gram positivas isoladas de material clínico humano, amostras ambientais e intestino do inseto. As espécies bacterianas incluíram Aeromonas hydrophila, A. culicicola, Morganella morganii subsp sibonii, Ochrobactrum anthropi, Weissella confusa, Escherichia coli, Ochrobactrum sp, Serratia sp, Kestersia sp, Ignatzschineria sp e Bacillus sp. A cepa Myroides sp foi resistente à penicilina G, eritromicina, estreptomicina, amicacina, canamicina, gentamicina, ampicilina, trimetoprim e tobramicina. Esta cepa apresentou atividade antimicrobiana contra todas as cepas exceto W.confusa, Ignatzschineria sp, A. hydrophila e M. morgani subsp sibonii. A resistência múltipla da cepa foi semelhante à de isolados clínicos, inibindo bactérias das amostras clínicas, ambientais e do intestino do inseto. Os genes de metalo beta lactamases (TUS/MUS estavam ausentes, excluindo-se a resistência mediada por esses genes, o que indica o envolvimento de um mecanismo alternativo de secreção.

  16. Extended-spectrum beta-lactamases in Klebsiella spp and Escherichia coli obtained in a Brazilian teaching hospital: detection, prevalence and molecular typing beta-lactamases de espectro ampliado em Klebsiella spp e em Escherichia coli obtidas em um hospital escola brasileiro: detecção, prevalência e tipagem molecular

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    Ana Lúcia Peixoto de Freitas

    2003-12-01

    Full Text Available His study was performed to compare the methods of detection and to estimate the prevalence of extended-spectrum beta-lactamases (ESBL among Klebsiella spp and E.coli in a university hospital in southern Brazil. We also used a molecular typing method to evaluate the genetic correlation between isolates of ESBL K.pneumoniae. Production of ESBL was investigated in 95 clinical isolates of Klebsiella spp and Escherichia coli from Hospital de Clínicas de Porto Alegre, using Kirby-Bauer zone diameter (KB, double-disk diffusion (DD, breakpoint for ceftazidime (MIC CAZ, increased zone diameter with clavulanate (CAZ/CAC and ratio of ceftazidime MIC/ceftazidime-clavulanate MIC (MIC CAZ/CAC. Molecular typing was performed by DNA macrorestriction analysis followed by pulsed-field gel electrophoresis. The KB method displayed the highest rates of ESBL (up to 70% of Klebsiella and 59% of E.coli, contrasting with all the other methods (p Este estudo foi desenvolvido para comparar métodos de detecção e para estimar a prevalência de Klebsiella spp e E.coli produtoras de beta-lactamases de espetro ampliado (ESBL em um Hospital Universitário no sul do Brasil. A correlação genética, determinada através de método molecular de tipagem, entre as amostras de K. pneumoniae também foi determinada. A produção de ESBL foi investigada em 95 amostras de Klebsiella spp e E.coli obtidas de pacientes no Hospital de Clínicas de Porto Alegre usando-se: medida do diâmetro a zona de inibição (KB, dupla-difusão de disco (DD, valores de concentração inibitória mínima da ceftazidima (MIC CAZ, aumento do diâmetro da zona de inibição com adição de clavulanato (CAZ/CAC e a relação entre o MIC da ceftazidima/MIC ceftazidima com clavulanato (MIC CAZ/CAC. A tipagem molecular foi realizada utilizando-se o método de macrorestrição de DNA e eletroforese em campo pulsado (PFGE. O método KB apresentou as maiores taxas de produção de ESBL (> 70% para Klebsiella e

  17. Improved detection of extended spectrum beta-lactamase (ESBL-producing Escherichia coli in input and output samples of German biogas plants by a selective pre-enrichment procedure.

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    Thorsten Schauss

    Full Text Available The presence of extended-spectrum beta-lactamase (ESBL-producing Escherichia coli was investigated in input (manure from livestock husbandry and output samples of six German biogas plants in 2012 (one sampling per biogas plant and two German biogas plants investigated in an annual cycle four times in 2013/2014. ESBL-producing Escherichia coli were cultured by direct plating on CHROMagar ESBL from input samples in the range of 100 to 104 colony forming units (CFU per g dry weight but not from output sample. This initially indicated a complete elimination of ESBL-producing E. coli by the biogas plant process. Detected non target bacteria were assigned to the genera Acinetobacter, Pseudomonas, Bordetella, Achromobacter, Castellaniella, and Ochrobactrum. A selective pre-enrichment procedure increased the detection efficiency of ESBL-producing E. coli in input samples and enabled the detection in five of eight analyzed output samples. In total 119 ESBL-producing E. coli were isolated from input and 46 from output samples. Most of the E. coli isolates carried CTX-M-type and/or TEM-type beta lactamases (94%, few SHV-type beta lactamase (6%. Sixty-four blaCTX-M genes were characterized more detailed and assigned mainly to CTX-M-groups 1 (85% and 9 (13%, and one to group 2. Phylogenetic grouping of 80 E. coli isolates showed that most were assigned to group A (71% and B1 (27%, only one to group D (2%. Genomic fingerprinting and multilocus sequence typing (MLST showed a high clonal diversity with 41 BOX-types and 19 ST-types. The two most common ST-types were ST410 and ST1210. Antimicrobial susceptibility testing of 46 selected ESBL-producing E. coli revealed that several isolates were additionally resistant to other veterinary relevant antibiotics and some grew on CHROMagar STEC but shiga-like toxine (SLT genes were not detected. Resistance to carbapenems was not detected. In summary the study showed for the first time the presence of ESBL-producing E

  18. Laboratory tests in the detection of extended spectrum beta-lactamase production: National Committee for Clinical Laboratory Standards (NCCLS screening test, the E-test, the double disk confirmatory test, and cefoxitin susceptibility testing

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    Pedro A. d'Azevedo

    2004-10-01

    Full Text Available Extended spectrum beta-lactamase (ESBL production by Klebsiella sp. and E. coli is an emerging problem. In this study, 107 clinical isolates (53 E. coli, 47 K. pneumoniae and 7 K. oxytoca screened as ESBL producers by the NCCLS disk diffusion procedure were submitted to a double disk confirmatory test (DDT and to the E-test double strip for confirmation of ESBL production by demonstration of clavulanic acid inhibition effect (CAIE. Only 72/107 (67% of the isolates were confirmed as ESBL producers by DDT, with diverse results among species. By the E-test, 58/107 (54% isolates were confirmed as ESBL producers, and 18/107 (17% were not determinable. Susceptibility to cefoxitin was found in 57/68 (83% of strains that did not show CAIE. ESBL detection remains a controversial issue and clinical laboratories are in need of a simple and effective way to recognize strains with this kind of resistance.

  19. Analysis of the beta-lactamase plasmid of borderline methicillin-susceptible Staphylococcus aureus: focus on bla complex genes and cadmium resistance determinants cadD and cadX.

    Science.gov (United States)

    Massidda, Orietta; Mingoia, Marina; Fadda, Daniela; Whalen, Michael B; Montanari, Maria Pia; Varaldo, Pietro E

    2006-03-01

    Borderline methicillin-susceptible Staphylococcus aureus strains are a rather homogeneous group, characterized by MICs of penicillinase-resistant penicillins (PRPs) at or just below the susceptibility breakpoint. Other features unique to this group include the presence of a pBW15-like beta-lactamase plasmid, the association with phage complex 94/96, and the production of a PRP-hydrolyzing beta-lactamase activity in addition to the classical penicillinase activity. The four HindIII fragments of pBORa53, a pBW15-like plasmid from the well-studied borderline S. aureus strain a53, were cloned in Escherichia coli, sequenced and analyzed. The plasmid (17,334 bp in size) contains 14 open reading frames (ORFs) and a complete copy of transposon Tn552, which harbors the three genes of the bla complex (blaZ, blaR1, and blaI) necessary for penicillinase production. Among the other 11 ORFs identified, two were homologous to cadmium resistance determinants of Staphylococcus lugdunensis and to the cadD and cadX genes recently detected in S. aureus. Consistent with this, strain a53 was found to be cadmium resistant. From a collection of 30 S. aureus isolates with borderline PRP MIC levels, 27 matched strain a53 in the positive amplification reactions with all of the four primer pairs targeting the cadD-cadX region, the presence of the 17.3-kb plasmid, and the level of cadmium resistance. The well-established S. aureus laboratory strain ATCC 29213 was also found to express cadD-cadX-mediated cadmium resistance. pBORa53 could be re-isolated from transformants obtained by transferring it into a PRP-susceptible recipient. However, while the transformants demonstrated levels of cadmium and penicillin resistance similar to those of strain a53, they remained fully susceptible to PRPs. PMID:16229889

  20. SHV-type extended-spectrum beta-lactamase (ESBL) are encoded in related plasmids from enterobacteria clinical isolates from Mexico beta-Lactamasas de espectro extendido (BLEE) tipo SHV están codificadas en plásmidos relacionados en aislamientos clínicos de enterobacterias en México

    OpenAIRE

    Ulises Garza-Ramos; Esperanza Martínez-Romero; Jesús Silva-Sánchez

    2007-01-01

    OBJECTIVE: In this work we report the molecular characterization of beta-lactam antibiotics resistance conferred by genes contained in plasmids from enterobacteria producing extended-spectrum beta-lactamases (ESBL). MATERIAL AND METHODS: Fourteen enterobacterial clinical isolates selected from a group of strains obtained from seven different hospitals in Mexico during 1990-1992 and 1996-1998 were analyzed at the Bacterial Resistance Laboratory (National Institute Public Health, Cuernavaca). M...

  1. Abattoirs as non-hospital source of extended spectrum beta lactamase producers: confirmed by the double disc synergy test and characterized by matrix-assisted laser desorption/ionization time of flight mass spectrometry.

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    Moses Nkechukwu Ikegbunam

    Full Text Available In this study, the presence of extended spectrum beta lactamase (ESBL producing organisms in abattoirs, a non-hospital community was investigated. The presence of ESBL-producing phenotypes was confirmed by the Double Disc Synergy Test (DDST. Out of the 99 isolates screened for ESBL, 28 (28.3% were confirmed positive. The positive isolates were characterised by using Matrix-Assisted Laser Desorption/Ionization Time of flight Mass Spectrometry. 50% of the isolates were Pseudomonas spp., the rest were different species of Acinetobacter, Stenotrophomonas and Achromobacter. Pseudomonas monteilli and Pseudomonas putida were the most occurring in the intestine. The entire positive ESBL producers were subjected to plasmid curing to ascertain the location of the resistant marker. The result of the plasmid curing indicated that the resistant genes were chromosomally borne. The findings have therefore established the presence of ESBL producing organisms in the gut of animals from abattoirs and the table were the meat are sold, and its rate of occurrence is comparable to hospital ICUs. Abattoir communities could probably be a source of human infection with ESBL expressing pathogens and possible transfer to non-ESBL producers.

  2. Variations in the Produce-Associated Microbiota and the Occurrence Frequency of Extended-Spectrum Beta-Lactamase Gram-Negative Bacteria Result in Different Level of Ingestion Risks

    KAUST Repository

    Bokhari, Osama

    2016-04-01

    A monitoring effort that spanned across one and a half years was conducted to examine three types of produce-associated microbiota. Produce type was determined to be the predominant factor affecting the microbial communities. Other significant factors that resulted in differences in the microbial populations were the origin and sampling date. Specifically, produce-associated microbiota among lettuce and tomatoes clustered based on the sampling period. Through molecular and cultivation-based approaches, sporadic presence of extended spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae and Acinetobacter baumannii was detected on lettuce and cucumbers during certain periods of sampling. Quantitative microbial risk assessment denoted varying levels of ingestion risks associated with different types of produce. In particular, the risks arising from ESBL-positive K. pneumoniae in the lettuce were higher than the acceptable annual risk of 10-4. Commonly used approaches to clean and wash the produce were insufficient in removing majority of the produce-associated microbiota. More invasive cleaning approaches or thorough cooking of the produce would be required to mitigate the associated risks. Most of the current reports of ESBL-positive bacterial isolates were identified in nosocomial environment. However, the carriage of such drug-resistant bacteria in food that is consumed daily

  3. Fenótipos de betalactamases em Klebsiella pneumoniae de hospital de emergência de Porto Alegre Phenotypes of beta-lactamases in Klebsiella pneumoniae from emergency hospital of Porto Alegre

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    Gabriela Meyer

    2011-02-01

    nosocomial infections raises the studies of bacterial resistance. Klebsiella pneumoniae is a Gram-negative bacteria most prevalent in the nosocomial environment, capable of expressing a variety of resistance enzymes, justifying their continued monitoring. OBJECTIVE: We sought to determine the frequency of beta-lactamases ESBL, AmpC plasmid, MBL and KPC in K. pneumoniae strains from emergency hospital in Porto Alegre (RS. MATERIAL AND METHOD: Fifty-eight bacterial isolates K. pneumoniae with reduced susceptibility to third generation cephalosporins and/or cefoxitin were included in this study. Phenotypic detection of beta-lactamases was carried out by using specific tests for ESBL, AmpC plasmid, KPC and MBL. RESULTS: Were detected a high frequency of ESBL (48.3%, followed by the AmpC plasmid/ESBL (15.5%, and two carbapenemase strains. DISCUSSION: The high rates of ESBL and ESBL/AmpC leads to the therapeutic use of carbapenens. Consequently, the massive use of carbapenens creates selective pressure favoring the more resistant strains, such as carbapenemase producing (KPC and/or MBL. The carbapenemase is very relevant due to the inherent therapeutic limitation, as well as their potential to spread. The resistance to all beta-lactam drugs used in the screening by disk diffusion can be a "flag" laboratory for presence of relevant mechanisms of resistance. CONCLUSION: Research of resistance phenotypes in Klebsiella is easy and feasible tool in microbiology laboratories, and is fundamental to understanding the local epidemiology of beta-lactamases, allowing the appropriateness of antimicrobial therapy more convenient.

  4. Caracterización de Klebsiella pneumoniae productora de la beta-lactamasa SHV-5, en una unidad de cuidados intensivos Characterization of SHV-5 beta-lactamase-producing Klebsiella pneumoniae in an intensive care unit

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    Verónica Andrade

    2004-12-01

    chain reaction (RAPD-PCR and serotyping, beta-Lactamase isoelectric focusing (IEF, and nucleotide sequencing of PCR products. RESULTS: Serotype 61 was predominant and correlated with genomic fingerprints of RAPD and PFGE in 11 of 15 isolates. One SHV-5-producer predominant clone with a high case-fatality rate was identified. CONCLUSIONS: Molecular biology techniques are useful tools to characterize the K. pneumoniae clone isolated from patients and health care workers, suggesting potential cross-transmission. These data call for strengthening control programs to prevent dissemination of nosocomial infections in the studied hospital.

  5. Métodos alternativos para detecção de betalactamase de espectro estendido em Escherichia coli e Klebsiella pneumoniae Alternative methods for the detection of extended-spectrum-beta-lactamase in Escherichia coli and Klebsiella pneumoniae

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    Alexsander Costa Martins

    2011-08-01

    Full Text Available INTRODUÇÃO: A resistência a antimicrobianos tem aumentado rapidamente nos últimos anos no Brasil e no mundo e, embora exista uma variedade de mecanismos de resistência, destaca-se a produção de betalactamase de espectro estendido (ESBL como um dos principais. Essas enzimas são mediadas por plasmídios, conferem resistência a vários antimicrobianos betalactâmicos e são inibidas por compostos, como ácido clavulânico, sulbactam e tazobactam. OBJETIVO: O objetivo deste estudo foi comparar metodologias alternativas à técnica padrão preconizada pelo Clinical and Laboratory Standards Institute (CLSI para detecção de ESBL. MATERIAL E MÉTODO: Foram realizados testes com 36 isolados (26 de E. coli e 10 de K. pneumoniae mediante disco combinado (CLSI e técnicas alternativas designadas meio disco (MD e substituição de discos (SD. CONCLUSÃO: As duas metodologias propostas apresentaram resultados satisfatórios com sensibilidade superior a 90% e custo inferior à técnica de referência (disco combinado, podendo ser utilizadas na pesquisa de ESBL.INTRODUCTION: Antimicrobial resistance has increased apace in Brazil and worldwide in the last years, even though there is a great variety of resistance mechanisms and extended spectrum beta-lactamases (ESBL is among the main ones. These enzymes are plasmid mediated, which causes resistance to some beta-lactam antimicrobials and are inhibited by compounds such as clavulanic acid, sulbactam and tazobactam. OBJECTIVE: The objective of this study was to compare alternative methods to the standard ESBL detection technique recommended by the Clinical and Laboratory Standards Institute (CLSI. MATERIAL AND METHOD: Tests with 36 isolates (26 E. coli and 10 K. pneumoniae were performed by means of CLSI disk diffusion method and alternative techniques designated as half disk (HD and disk substitution (SD. CONCLUSION: Both methods yielded satisfactory results with higher sensitivity (90% and lower costs

  6. Evaluation of the contemporary occurrence rates of metallo-beta-lactamases in multidrug-resistant Gram-negative bacilli in Japan: report from the SENTRY Antimicrobial Surveillance Program (1998-2002).

    Science.gov (United States)

    Jones, Ronald N; Deshpande, Lalitagauri M; Bell, Jan M; Turnidge, John D; Kohno, Shigeru; Hirakata, Yoichi; Ono, Yasuo; Miyazawa, Yukihisa; Kawakama, Sayoko; Inoue, Matsuhisa; Hirata, Yasuyoshi; Toleman, Mark A

    2004-08-01

    Metallo-beta-lactamases (M beta L) were initially characterized in Japan, usually of the IMP-type, and found in Pseudomonas aeruginosa (PSA), Acinetobacter spp. (ACB), or Serratia marcescens (SM). The number of M beta L types has increased worldwide, but geographic dissemination within Japan has appeared limited. This study compares baseline levels of M beta L resistance from two 22-center studies (1996-1997) to the longitudinal sample (3 sites) of Japanese isolates from the SENTRY Antimicrobial Surveillance Program (1998-2002). All minimal inhibitory concentration results were determined by reference methods. A total of 26.8% PSA, 3.4% ACB, and 3.1% Enterobacteriaceae (enterobacters and SM) with resistance to monitored carbapenems (CARB) (minimal inhibitory concentration, > or =8 microg/mL) were screened for M beta L production by disk approximation tests (EDTA and 2-MPA inhibitors), CARB hydrolysis by enzyme extracts, and selected PCR primers for known M beta L types. All M beta L-positive strains (10) were sequenced to determine enzyme identification. Clonality in each center was determined by automated ribotyping and PFGE. The CARB susceptibility rates in PSA decreased (80.7% to 62.0%) over the monitored interval (1998-2002), but varied by medical center location. Among CARB-resistant isolates, 10.8% were attributed to M beta L strains (1.1% of all PSA tested). M beta L identification showed the following: five PSA (three IMP-1, two IMP-2), four SM (one IMP-1, two IMP-1 + OXA-1, and one IMP-11). Also a single ACB had an IMP-1. Eight of 10 M beta L isolations occurred between 2000 and 2002; four occurred in 2002. BRL42715, an AMP-C inhibitor, confirmed AMP-C-mediated resistance in 87.3% of PSA, and outer membrane protein changes were also discovered by membrane studies. Prior results (22 sites, 1997-1998) showed CARB resistance at 22.4-25.6% and 0.5-0.9% M beta Ls (IMP-1) overall; it was slightly elevated in this SENTRY Program sample. In conclusion, M beta L

  7. Epidemiologia das betalactamases de espectro estendido no Brasil: impacto clínico e implicações para o agronegócio Epidemiology of extended-spectrum beta-lactamases in Brazil: clinical impact and implications for agribusiness

    Directory of Open Access Journals (Sweden)

    Ketrin Cristina da Silva

    2012-04-01

    Full Text Available A emergência e a disseminação de bactérias produtoras de betalactamases de espectro estendido (ESBL têm sido retratadas como grande problema de saúde pública, especialmente no que diz respeito a patógenos associados às infecções relacionadas com a assistência à saúde (IRAS. No Brasil, a maior preocupação inclui as altas taxas de resistência a Klebsiella pneumoniae e Escherichia coli, embora as ESBL estejam amplamente disseminadas entre os membros da família Enterobacteriaceae e sejam descritas como enzimas do tipo TEM, SHV, CTX-M, VEB, BES e GES em diferentes estados. Contudo, as enzimas dos grupos CTX-M-2, CTX-M-8 e CTX-M-9 são as mais prevalentes em território brasileiro. Além do ambiente hospitalar, as ESBL de origem comunitária e ambiental têm sido retratadas. A CTX-M-2 também tem sido identificada em Salmonella, oriunda do ciclo de produção animal, o que é alarmante para o Brasil diante da importância que a exportação de carnes assume para o agronegócio. Dessa forma, faz-se necessária a regulamentação do uso de antimicrobianos em todos os setores, a fim de evitar a disseminação de bactérias resistentes e resguardar a qualidade e a inocuidade dos alimentos. Portanto, o presente estudo visa retratar o panorama geral da epidemiologia das ESBL no Brasil, enfatizando o impacto clínico, ambiental e econômico.The emergence and dissemination of extended-spectrum beta-lactamase bacteria (ESBL have been reported as a major public health issue, mainly with regard to nosocomial infections. In Brazil, ESBL are widely disseminated among the Enterobacteriaceae family and enzymes TEM, SHV, CTX-M, VEB, BES and GES have been reported in several states. However, the major concern is the high rates of resistant Klebsiella pneumoniae and Escherichia coli strains. Currently, ESBL belonging to CTX-M-2, CTX-M-8 and CTX-M-9 subtypes are the most prevalent in Brazil. Apart from nosocomial infections, ESBL bacteria from

  8. Avaliação da atividade in vitro do meropenem contra cepas de Klebsiella pneumoniae produtoras de betalactamases de espectro expandido isoladas na cidade de Fortaleza, Ceará Evaluation of the in vitro activity of meropenem against strains of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae isolated in the city of Fortaleza, Ceará

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    Everardo Albuquerque Menezes

    2007-06-01

    Full Text Available Betalactamases de espectro estendido são mediadas por plasmídios. Essas enzimas possuem a habilidade de hidrolizar antibióticos beta-lactâmicos. Nesse estudo, avaliamos a atividade in vitro do meropenem contra cepas de Klebsiella pneumoniae produtoras de ESBLs. Foram estudadas 14 cepas. A susceptibilidade dessas cepas para o meropenem foi de 100%.Extended-spectrum beta-lactamases are plasmid-mediated. These enzymes have the ability to hydrolyze beta-lactam antibiotics. In this study, we evaluated the in vitro activity of meropenem against ESBL-producing Klebsiella pneumoniae strains. Fourteen strains were studied. The susceptibility of these strains to meropenem was 100%.

  9. SHV-type extended-spectrum beta-lactamase (ESBL are encoded in related plasmids from enterobacteria clinical isolates from Mexico beta-Lactamasas de espectro extendido (BLEE tipo SHV están codificadas en plásmidos relacionados en aislamientos clínicos de enterobacterias en México

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    Ulises Garza-Ramos

    2007-12-01

    Full Text Available OBJECTIVE: In this work we report the molecular characterization of beta-lactam antibiotics resistance conferred by genes contained in plasmids from enterobacteria producing extended-spectrum beta-lactamases (ESBL. MATERIAL AND METHODS: Fourteen enterobacterial clinical isolates selected from a group of strains obtained from seven different hospitals in Mexico during 1990-1992 and 1996-1998 were analyzed at the Bacterial Resistance Laboratory (National Institute Public Health, Cuernavaca. Molecular characterization included PFGE, IEF of beta-lactamases, bacterial conjugation, PCR amplification and DNA sequencing, plasmid extraction and restriction. RESULTS: Isolates were genetically unrelated. ESBL identified were SHV-2 (5/14 and SHV-5 (9/14 type. Cephalosporin-resistance was transferable in 9 of 14 (64% clinical isolates with only one conjugative plasmid, DNA finger printing showed a similar band pattern in plasmids. CONCLUSIONS: The dissemination of cephalosporin resistance was due to related plasmids carrying the ESBL genes.OBJETIVO: En este trabajo se reporta la caracterización molecular de la resistencia a antibiótico beta-lactámicos conferida por genes contenidos en plásmidos de enterobacterias productoras de beta-lactamasas de espectro extendido (BLEEs. MATERIAL Y MÉTODOS: Catorce aislamientos clínicos de enterobacterias fueron seleccionados por conveniencia de un banco de cepas obtenidas de siete diferentes hospitales de México durante los periodos 1990-1992 y 1996-1998 y fueron procesados en el Laboratorio de Resistencia Bacteriana (Instituto Nacional de Salud Pública, Cuernavaca. En la caracterización se empleó PFGE, IEF para beta-lactamasas, conjugación bacteriana, amplificación por PCR y secuenciación de DNA, extracción y restricción de plásmidos. RESULTADOS: Las 14 cepas fueron no relacionadas genéticamente. Se identificaron BLEEs tipo SHV-2 (5/14 y SHV-5 (9/14. La resistencia a cefalosporinas fue transferida por

  10. Study on Acinetobacter baumannii plasmid with 3 types of beta-lactamase genes in a burn ward%关于携带3种β内酰胺酶基因的鲍氏不动杆菌质粒的研究

    Institute of Scientific and Technical Information of China (English)

    李蓉; 李文林; 石小玉; 曾元临; 徐小文; 赵林

    2008-01-01

    Objective To study the transferrable character of Aciuetobacter baumanni i(AB)plasraids with 3 types of beta-lactamase gene. Methods The plasmid of multi-drug resistant AB(donor)isolated from burn wound were transferred to E.coil ATCC25922 (receptor) through conjugation,and drug sensitivity was also observed.Drug-resistant gene and Stability of filial generation and zygote were analyzed by PCR. Results The dug-resistance of donor plasmids to Sulfamethoxazole, Ampicillin, Cefalotin, Cetpodoxime,Cefuroxime,Imipenem/Cilastatin and Ampicillin/SuIbactam,and three types of beta-lactamase gene were transferred to the receptor,and were also stably transmitted for passages. The minimum inhibitor concentration ot receptor to Sulfamethoxazole was>2 mg/L after conjugation with donor,and inhibitory character could be transferred to next generation. Conclusion blaTEM-1,blaPER-1 and blaOXA-23 genes carried in the ptasmid of AB can be transferred through conjugation and stably transmitted for passages,and it is one of the molecular mechanisms for AB with multi-drug resistance after burn infections.%目的 了解携带3种β内酰胺酶基因的鲍氏不动杆菌质粒的町传递性.方法 选取从烧伤创面分离出的多重耐药鲍氏不动杆菌(供体菌),将之与大肠埃希菌ATCC 25922(受体菌)进行耐药质粒接合、药物敏感试验,并采用PCR分析接合子及其子代的耐药基因型、传代稳定性. 结果鲍氏不动杆菌通过接合将其携带对磺胺甲恶唑、氨苄西林、头孢噻吩、头孢博肟、头孢呋辛、亚胺培南/两司他丁和氨苄西林/舒巴坦的耐药性质粒及3种耐药基因传递给受体菌(例如经接合,使受体菌对磺胺甲恶唑的最低抑菌浓度>2 mg/L),且可稳定传代. 结论鲍氏不动杆菌质粒上携带可接合传递并稳定传代的β内酰胺酶基冈(blaTEM-1、blaPER-1、blaOXAS-23),是烧伤感染后其具有多重耐药性的分子生物学机制之一.

  11. Antibacterial activity of ceftizoxime, a beta-lactamase-stable cephalosporin.

    OpenAIRE

    Fu, K P; Neu, H C

    1980-01-01

    The in vitro activity of ceftizoxime was compared with that of other beta-lactam antibiotics against 538 isolates. Ceftizoxime was the most active agent tested against Escherichia coli and Klebsiella, inhibiting 80% at 0.025 microgram/ml. It was more active than cefotaxime against Enterobacter cloacae and E. aerogenes. Ceftizoxime was more active than cefoxitin, cefotaxime, cefoperazone, and carbenicillin against Proteus mirabilis and indole-positive Proteus. It inhibited 97% of multiresistan...

  12. [Extended spectrum beta-lactamases in Danish Klebsiella isolates].

    Science.gov (United States)

    Hansen, D S; Sirot, D; Kolmos, H J

    1998-04-01

    This study presents the first two cases of infections with Klebsiella pneumoniae producing extended spectrum betalactamases (ESBL) that have been recorded in Denmark. They presented as a urinary tract infection and a generalized infection in a patient admitted to an intensive care unit. Both patients had been treated with broad spectrum antibiotics prior to infection. Presumably, one of the strains had been imported from Turkey. The ESBL of the two strains were characterized as SHV-2 and SHV-5, respectively. Patients transferred from hospitals abroad should be screened for Klebsiella producing ESBL, in addition to MRSA and other multiresistant organisms. A restrictive antibiotic policy and strict hygienic precautions are essential measures to control the selection and spread of such organisms in the hospital environment. PMID:9599523

  13. Mortalidad por bacteriemia causada por Escherichia coli y Klebsiella spp. productoras de beta lactamasas de espectro extendido: cohorte retrospectiva en un hospital de Lima, Perú Mortality caused by bacteremia Escherichia coli and Klebsiella spp. extended-spectrum beta-lactamase- producers: a retrospective cohort from a hospital in Lima, Peru

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    Diego Adrianzén

    2013-03-01

    Full Text Available Objetivos. Evaluar los factores asociados a la mortalidad causada por bacteriemias por Escherichia coli y Klebsiella spp. productoras de beta lactamasas de espectro extendido (BLEE. Materiales y Métodos. Se realizó un estudio de cohortes retrospectivo, que incluyó 85 pacientes mayores de 16 años con diagnóstico de bacteriemia por Escherichia coli o Klebsiella spp. hospitalizados entre 2006 y 2008 en el Hospital Nacional Cayetano Heredia. Las cohortes se clasificaron de acuerdo a la producción de BLEE según los resultados de los hemocultivos. Se evaluaron los factores asociados a la mortalidad cruda y atribuible empleando regresión de Poisson en un modelo multivariado, con lo que se obtuvo riesgos relativos ajustados (RRa. Además, se construyeron curvas de mortalidad. Resultados. Se encontró que el 35,3% de las bacteriemias fueron debidas a cepas productoras de BLEE. El análisis de la mortalidad cruda mostró una mayor mortalidad en el grupo de cepas productoras de BLEE (63,3%. El RRa fue de 1,5 (IC95%: 1,02-2,3. En el caso de mortalidad atribuible, la proporción también fue mayor (63,3%, el RRa fue de 1,9 (IC95%: 1,2-2,9. El uso de catéter venoso central fue otro factor asociado tanto a la mortalidad cruda (RRa= 2,4; IC95%: 1,2- 4,8 como a la mortalidad atribuible (RRa= 3,8; IC95%: 1,6-8,8. Conclusiones. La producción de BLEE es un factor de riesgo independiente para mortalidad por bacteriemia causada por E. coli y Klebsiella spp. Su presencia debe evaluarse tras la sospecha diagnóstica y la elaboración de la terapéutica inicial, lo que podría disminuir la mortalidad por esta causaObjectives. To evaluate the factors associated to mortality caused by bacteremia due to Escherichia coli and Klebsiella spp. producers of extended-spectrum beta-lactamase (ESBL. Materials and methods. We performed a retrospective cohort study, including 85 patients older than 16 and diagnosed with bacteremia by Escherichia coli or Klebsiella spp

  14. Avaliação da acurácia de testes laboratoriais para detecção de amostras de Klebsiella pneumoniae produtora de betalactamase de espectro estendido Comparison of different methods for detection of Klebsiella pneumoniae isolates producers of extended spectrum beta-lactamase

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    Andrea dos Santos Pereira

    2003-01-01

    spectrum beta-lactamases (ESBL represent a common resistance problem among Brazilian hospitals. Due to the difficulty of ESBL detection in the clinical laboratory, these bacterial isolates require a reproducible, efficient, and low cost detection method. The aim of the present study was to evaluate the efficacy of detection of K. pneumoniae ESBL isolates by two methods: the Etest ESBL strip and the inhibitor potentiated disk diffusion test with clavulanic acid (clavulanate-potentiation test. The sensitivity and the specificity of beta-lactam agents against these isolates were also evaluated. The experiments were performed on a total of 134 K. pneumoniae isolates recovered from blood specimens in our institution from July 1996 to July 2001. The samples were tested for ESBL production by the NCCLS screen test, clavulanate-potentiation test and Etest ESBL strip. Isolates presenting positive results for the screen test and for at least one of the evaluated tests were considered ESBL producers (gold standard. The results of this study yielded a 100% specificity and sensitivity for the clavulanate-potentiation test, and the best indicators of ESBL production were cefotaxime and cefpodoxime. The Etest ESBL strip also turned out to be a very sensitive (96% and specific (100% method, being cefotaxime the most efficient substrate. According to the results of this investigation, the clavulanate potentiation disk diffusion test displayed an excellent performance and can be easily implemented in routine clinical laboratories as a practical, reliable, and accurate method.

  15. Beta-lactamases among Extended spectrum Beta-lactamase resistant (ESBL) Salmonella from poultry, poultry products and human patients in The Netherlands

    DEFF Research Database (Denmark)

    Hasman, Henrik; Mevius, D.; Veldman, K.; Olesen, I.; Aarestrup, Frank Møller

    2006-01-01

    enterica serovars Blockley, Thomson, London, Enteritidis phage type 14b, Paratyphi B, Virchow and Typhimurium phage types 11 and 507. We also found the bla(TEM-20) gene in S. Paratyphi B var. Java and the bla(TEM-63) gene in S. Isangi. Furthermore, we detected the bla(CTX-M-28) gene in S. Isangi and the...... bla(CTX-M-3) gene in S. Typhimurium phage type 507. The bla(CTX-M-2) gene was identified in S. Virchow, which also contained a copy of the bla(SHV-2) gene and a copy of the bla(TEM-1) gene. The bla(SHV-12) gene was found alone in S. Concord and together with the bla(TEM-52) gene in S. Typhimurium...... purification, restriction fragment length polymorphism (RFLP) and pulsed-field gel electrophoresis (PFGE) were employed to further characterize a subset of the isolates. Results: A great genetic diversity was seen among the isolates. The bla(TEM-52) gene was most predominant and was found among Salmonella...

  16. Metallo-beta-lactamases among imipenem-resistant Pseudomonas aeruginosa in a Brazilian university hospital

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    Maria Renata Gomes Franco

    2010-01-01

    Full Text Available INTRODUCTION: Imipenem-resistant Pseudomonas aeruginosa resulting from metallo-β-lactamases has been reported to be an important cause of nosocomial infection and is a critical therapeutic problem worldwide, especially in the case of bacteremia. OBJECTIVES: To determine the frequency of metallo-β-lactamases among imipenem-resistant Pseudomonas aeruginosa isolates and to compare methods of phenotypic and molecular detection. METHODS: During 2006, 69 imipenem-resistant Pseudomonas aeruginosa samples were isolated from blood and tested for metallo-β-lactamase production using both phenotypic methods. Minimal Inhibitory Concentratrions (MIC (μg/mL was determined with commercial microdilution panels. Pulsed Field Gel Electrophoresis (PFGE was performed among metallo-β-lactamase producers. RESULTS: Of all the blood isolates, 34.5% were found to be imipenem-resistant Pseudomonas aeruginosa. Positive phenotypic tests for metallo-β-lactamases ranged from 28%-77%, and Polymerase Chain Reaction (PCR were positive in 30% (of note, 81% of those samples were blaSPM-1 and 19% were blaVIM-2. Ethylenediamine tetracetic acid (EDTA combinations for the detected enzymes had low kappa values; thus, care should be taken when use it as a phenotypic indicator of MBL. Despite a very resistant antibiogram, four isolates demonstrated the worrisome finding of a colistin MIC in the resistant range. PFGE showed a clonal pattern. CONCLUSION: Metallo-β-lactamases among imipenem-resistant Pseudomonas aeruginosa were detected in 30.4% of imipenem-resistant Pseudomonas aeruginosa isolates. This number might have been higher if other genes were included. SPM-1 was the predominant enzyme found. Phenotypic tests with low kappa values could be misleading when testing for metallo-β-lactamases. Polymerase Chain Reaction detection remains the gold standard.

  17. Prevalence of extended spectrum beta lactamases among strains of Pseudomonas aeruginosa isolated from burn patients

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    Mirsalehian

    2008-08-01

    Full Text Available Background: The resistance of Pseudomonas aeruginosa strains to broad spectrum cephalosporins may be mediated by extended spectrum b-lactamases (ESBLs. These enzymes are encoded by different genes located either on chromosome or plasmids. In this study, we determined the antimicrobial resistance patterns of P. aeruginosa isolates and screened for ESBL production. Methods: After isolation from burn patients in Tehran Hospital, identification of P. aeruginosa isolates were assessed using biochemical tests. We then performed disk agar diffusion (DAD according to CLSI guidelines to determine the pattern of antimicrobial resistance. The frequency of ESBLs and prevalence of the OXA-10 and PER-1 genes were determined with combined disk and polymerase chain reaction (PCR methods, respectively. Results: One hundred strains of P. aeruginosa were isolated. The resistance of these strains to cephpodoxime, aztreonam, ciprofloxacin, ofloxacin, ceftazidime, cefepime, imipenem, meropenem, cefotaxime, levofloxacin, piperacilin- tazobactam and ceftriaxon was 100%, 90%, 83%, 92%, 85%, 88%, 63%, 66%, 98%, 89%, 70% and 91%, respectively. Of these, 40 strains (40% were ESBL positive, 29 strains (29% were OXA-10 positive and 18 strains (18% were PER-1 positive. Conclusion: Our results confirm the need for proper antimicrobial therapy in burn hospitals, considering the resistance pattern and frequency of strains producing ESBLs and the presence of the OXA-10 and PER-1 genes. Since an increase in the prevalence of ESBL in P. aeruginosa strains might lead to the transfer of these ESBL genes to other gram-negative bacteria, we recommend the use of appropriate drugs, especially cephalosporins, in burn hospitals.

  18. Prevalence of Extended-spectrum beta-lactamase Producing Enterobacteriaceae Strains in Latvia

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    Sandra Leja

    2007-10-01

    Full Text Available A total of 507 strains of the Enterobacteriaceae family were tested for the production of ESBL using mini API, ATB Expert system as a screening method, as well as the double disk method and E-test for confirmation. The prevalence of ESBL producing E. coli is 5.95%, Klebsiella spp. strains 37.7%. All ESBL- producing isolates are susceptible to imipenem and clavulanate. The susceptibility to other antimicrobials varies from 36 to 92%.

  19. Extended-spectrum beta-lactamase-producing enterobacteriaceae : epidemiology and dynamics of fecal carriage

    OpenAIRE

    Titelman, Emilia

    2013-01-01

    ESBL-producing Enterobacteriaceae (EPE) has become a major cause of community acquired urinary tract infection (UTI), and fecal carriage of EPE is emerging worldwide. The aims of this thesis were to study the molecular epidemiology of ESBL-enzymes in Stockholm (I) to evaluate treatment alternatives to the carbapenems for EPE (I-III), and to study the duration of fecal carriage and identify factors associated with prolonged carriage (IV). Paper I describes a consecutive collection of EPE...

  20. Beta-Lactamase Producing Escherichia coli Isolates in Imported and Locally Produced Chicken Meat from Ghana

    OpenAIRE

    Rasmussen, Mette Marie; Japheth A Opintan; Frimodt-Møller, Niels; Styrishave, Bjarne

    2015-01-01

    The use of antibiotics in food animals is of public health concern, because resistant zoonotic pathogens can be transmitted to humans. Furthermore, global trade with food may rapidly spread multi-resistant pathogens between countries and even continents. The purpose of the study was to investigate whether imported chicken meat and meat from locally reared chicken are potential sources for human exposure to multi resistant Escherichia coli isolates. 188 samples from imported and locally produc...

  1. Prevalence of extended-spectrum beta-lactamase-producing bacteria in food

    OpenAIRE

    Tham, Johan

    2012-01-01

    Johan Tham,1 Mats Walder,2 Eva Melander,2,3 Inga Odenholt11Infectious Diseases Unit, Department of Clinical Sciences, 2Medical Microbiology, Department of Laboratory Medicine, Lund University, Malmö, Sweden; 3Department of Infection Control, Laboratory Medicine, Skåne County, SwedenAbstract: Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae with Cefotaximase–München (CTX-M) enzymes are rapidly increasing worldwide and pose a threa...

  2. Metallo beta lactamase mediated resistance in Carbapenem resistant gram-negative bacilli: A cause for concern

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    Malini Jagannatha Rao, Shruti A Harle, Padmavathy M, Umapathy BL, Navaneeth BV

    2014-04-01

    Full Text Available Introduction: The emergence of acquired metallo-β-lactamases (MBL in Gram-negative bacilli is becoming a therapeutic challenge, as these enzymes usually possess a broad hydrolysis profile that includes carbapenems, extended-spectrum β-lactams. Aim: To detect Extended spectrum β-lactamases and metallo-β-lactamase in carbapenem resistant Gram negative clinical isolates from various clinical specimens and to evaluate their antibiotic susceptibility patterns. Material and Methods: A total of 100 non duplicates imipenem resistant isolates were tested for the presence of extended spectrum β-lactamases by phenotypic confirmatory test, metallo-β-lactamases by Double disk synergy test with various distances from edge to edge (10mm,15mm,20mm, between the IPM and EDTA and combined disc test. Result: Of the 100 IMP resistant isolates screened 30 (30% were MBL positive by phenotypic methods, i.e., double disk synergy test and combined disc test. Co-existence of Extended spectrum β-lactamases and MBL were detected in 3 (30%. All the 30 MBL positive isolates had shown synergy at (100% at 10 mm distance, 27 (90% isolates had shown synergy at 15 mm distance and 13 (43.4% isolates were shown synergy at 20 mm distance. All the 30 MBLs producers were multidrug resistant and 27 (90% were sensitive to colistin (CL. All MBL positive Pseudomonas aeruginosa were sensitive to polymyxin B (100µg. Conclusion: Microbiologists are now facing a challenge of drug resistance due to MBL production. Although CLSI guidelines do not quote about the ESBL detection in Pseudomonas aeruginosa MBLs and ESBL have to be detected in them. The use of combination tests would increase the sensitivity to detect the presence of MBL among the clinical isolates of Gram-negative bacilli. The spread of MBL producing Gram negative organism can be prevented if they are detected in all isolates and routinely adopted in all laboratories.

  3. Extended Spectrum Beta Lactamase producing Cephalosporin resistant Salmonella Typhi, reported from Rawalpindi, Pakistan.

    Science.gov (United States)

    Munir, Tehmina; Lodhi, Munir; Ansari, Jawad Khaliq; Andleeb, Saadia; Ahmed, Mushtaq

    2016-08-01

    Typhoid is endemic in many parts of southeast Asia. Due to the resistance of the organism to first line of antibiotics (ampicillin, chloramphenicol, cotrimoxazole) as well as to fluoroquinolones, third generation cephalosporins have been in use for the empiric treatment of typhoid for years. However an increasing incidence of Salmonella Typhi is being reported sporadically from various regions. We report a case of typhoid due to Salmonella Typhi which was non-responsive to treatment with a cephalosporin, was found to be multidrug resistant and resistant to ciprofloxacin and third generation cephalosporin as well. The patient was finally treated successfully with intravenous administration of a carbapenem. PMID:27524545

  4. Carriage of beta-lactamase-producing Enterobacteriaceae among nursing home residents in north Lebanon

    Directory of Open Access Journals (Sweden)

    Iman Dandachi

    2016-04-01

    Conclusions: The high prevalence of MDR Enterobacteriaceae detected in this study and the emergence of carbapenem resistance is alarming. Efficient infection control measures and antibiotic stewardship programs are urgently needed in these settings in order to limit the spread of resistant strains.

  5. Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae: Epidemiology, Risk Factors, and Duration of Carriage

    OpenAIRE

    Tham, Johan

    2012-01-01

    In human medicine, the most important family of bacteria is Enterobacteriaceae, which includes genera and species that cause well-defined diseases and also nosocomial infections. These bacteria can give rise to numerous infections such as septicaemia and urinary tract infections. It is important to combat these infections with effective antibiotics, because many of them are associated with high mortality rates if left untreated. In infections with ESBL-producing bacteria in low-endemic countr...

  6. Fecal Colonization with Extended-Spectrum Beta-Lactamase and AmpC-Producing Escherichia coli

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    Mohamed H. Al-Agamy

    2016-01-01

    Full Text Available Background. Extended-spectrum β-lactamases (ESβLs and AmpC β-lactamases cause β-lactam resistance in Escherichia coli. Fecal colonization by ESβL- and/or AmpC-positive E. coli is a source of nosocomial infections. Methods. In order to investigate inpatient fecal colonization by ESβLs and AmpC, antibiotic sensitivity tests were conducted and minimum inhibitory concentrations (MICs were determined using the disk diffusion method and E-test, respectively. Characterization of ESβL and AmpC was performed using E-test strips, and a set of PCRs and DNA sequence analyses were used to characterize the ESβL and AmpC genes. Results. The whole collection of E. coli isolates (n=50 was sensitive to imipenem, tigecycline, colistin, and fosfomycin, while 26% of the isolates showed reduced susceptibility to ceftazidime (MIC ≥ 4 μg/mL. ESβL was phenotypically identified in 26% (13/50 of cases, while AmpC activity was detected in two ESβL-producing E. coli isolates. All ESβL-producing E. coli were positive for the CTX-M gene, eleven isolates carried blaCTX-M-15, and two isolates carried blaCTX-M-14 gene. Two CTX-M-positive E. coli isolates carried blaCMY-2. Conclusions. The alimentary tract is a significant reservoir for ESβL- and/or AmpC-producing E. coli, which may lead to nosocomial infection.

  7. Fecal Colonization with Extended-Spectrum Beta-Lactamase and AmpC-Producing Escherichia coli.

    Science.gov (United States)

    Al-Agamy, Mohamed H; El Mahdy, Taghrid S; Shibl, Atef M

    2016-01-01

    Background. Extended-spectrum β-lactamases (ESβLs) and AmpC β-lactamases cause β-lactam resistance in Escherichia coli. Fecal colonization by ESβL- and/or AmpC-positive E. coli is a source of nosocomial infections. Methods. In order to investigate inpatient fecal colonization by ESβLs and AmpC, antibiotic sensitivity tests were conducted and minimum inhibitory concentrations (MICs) were determined using the disk diffusion method and E-test, respectively. Characterization of ESβL and AmpC was performed using E-test strips, and a set of PCRs and DNA sequence analyses were used to characterize the ESβL and AmpC genes. Results. The whole collection of E. coli isolates (n = 50) was sensitive to imipenem, tigecycline, colistin, and fosfomycin, while 26% of the isolates showed reduced susceptibility to ceftazidime (MIC ≥ 4 μg/mL). ESβL was phenotypically identified in 26% (13/50) of cases, while AmpC activity was detected in two ESβL-producing E. coli isolates. All ESβL-producing E. coli were positive for the CTX-M gene, eleven isolates carried bla CTX-M-15, and two isolates carried bla CTX-M-14 gene. Two CTX-M-positive E. coli isolates carried bla CMY-2. Conclusions. The alimentary tract is a significant reservoir for ESβL- and/or AmpC-producing E. coli, which may lead to nosocomial infection. PMID:27340657

  8. Carriage of ESBL (extended spectrum beta-lactamases) - producing bacteria - knowledge, emotional impact and risk assessment

    OpenAIRE

    Wiklund, Susanne

    2015-01-01

    Antibiotic-resistant bacteria, such as ESBL-producing Enterobacteriaceae, have become a growing public health threat. The overall aim of this thesis was to explore how the increasing prevalence of ESBL-producing bacteria affects different groups among the Swedish population. Paper I described the knowledge and understanding of antibiotic-resistant bacteria, and of the risk of becoming a carrier of such bacteria, among ninety-five Swedish travelers before travel to high-risk areas. A questionn...

  9. Emergence of VIM-2 metallo-beta-lactamase producing Ralstonia pickettii clinical isolate in India

    Directory of Open Access Journals (Sweden)

    A Khajuria

    2014-01-01

    Full Text Available A multidrug-resistant clinical isolate of Ralstonia pickettii from a woman was analysed. Modified Hodge test was positive for carbapenemase production. Conjugation experiment revealed the presence of conjugative plasmid of >140 Kb size typed as IncN type. This is the first report of emergence blaVIM-2 in R. pickettii in India.

  10. Audouin's gull, a potential vehicle of an extended spectrum beta-lactamase producing Salmonella Agona

    DEFF Research Database (Denmark)

    Antilles, Noelia; Garcia-Migura, Lourdes; Joensen, Katrine Grimstrup;

    2015-01-01

    The genome of a multidrug-resistant Salmonella Agona isolated from Larus audouinii (Audouin's gull) in Spain was examined. The isolate showed high levels of resistance to different antimicrobials, including third generation cephalosporins and fluoroquinolones, which is a public health concern as...... the presence of two plasmids. Migratory Audouin's gulls have the ability to cover long distances during annual movements. Therefore, they have the potential to disseminate multidrug-resistant Salmonella and resistance genes in the environment and over great geographic distances, contributing to the...

  11. Prevalence of bla CTX M extended spectrum beta lactamase gene in enterobacteriaceae from critical care patients

    Directory of Open Access Journals (Sweden)

    R Indra Priyadharsini

    2011-01-01

    Conclusions: Early detection of CTX M producing Enterobacteriaceae by continuous surveillance and thereby reducing their spread and restricted use of third generation Cephalosporins (3GC antibiotics could be the possible routes to prevent the emergence and spread of CTX M ESBL producing organisms.

  12. A practical approach for reliable detection of AmpC Beta-Lactamase producing Enterobacteriaceae

    OpenAIRE

    Polsfuss, S; Bloemberg, G V; Giger, J; Meyer, V.; Böttger, E.C.; Hombach, M

    2011-01-01

    In this prospective study all Enterobacteriaceae isolates (n=2129) recovered in the clinical microbiology laboratory during October 2009 to April 2010 were analyzed for AmpC production. CLSI cefoxitin and cefotetan susceptibility breakpoints and CLSI critical ESBL diameters were used to screen for potential AmpC producers. In total, 305 isolates (211 potential AmpC producers, 94 AmpC screen negative isolates as control group) were further analyzed by multiplex PCR for the detection of plasmid...

  13. Practical Approach for Reliable Detection of AmpC Beta-Lactamase-Producing Enterobacteriaceae ▿

    OpenAIRE

    Polsfuss, Silke; Bloemberg, Guido V.; Giger, Jacqueline; Meyer, Vera; Böttger, Erik C.; Hombach, Michael

    2011-01-01

    In this prospective study all Enterobacteriaceae isolates (n = 2,129) recovered in the clinical microbiology laboratory during October 2009 to April 2010 were analyzed for AmpC production. Clinical and Laboratory Standards Institute (CLSI) cefoxitin and cefotetan susceptibility breakpoints and CLSI critical ESBL diameters were used to screen for potential AmpC producers. In total, 305 isolates (211 potential AmpC producers and 94 AmpC screen-negative isolates as a control group) were further ...

  14. Ampc Beta Lactamases Among Gram Negative Clinical Isolates From A Tertiary Hospital, South India

    OpenAIRE

    Mohamudha, Parveen R.; Harish, B. N.; S C Parija

    2010-01-01

    AmpC β-lactamases are cephalosporinases that hydrolyze cephamycins as well as other extended-spectrum cephalosporins and are poorly inhibited by clavulanic acid. Although reported with increasing frequency, the true rate of occurrence of AmpC β-lactamases in different organisms, including members of Enterobacteriaceae, remains unknown. The present study was designed to determine the occurrence of AmpC enzyme-harbouring Gram-negative clinical isolates in a tertiary care hospital in P...

  15. Phenotypic detection of broad-spectrum beta-lactamases in microbiological practice

    OpenAIRE

    Sedlakova, Miroslava Htoutou; Hanulik, Vojtech; Chroma, Magdalena; Hricova, Kristyna; Kolar, Milan; Latal, Tomas; Schaumann, Reiner; Rodloff, Arne C.

    2011-01-01

    Summary Background Enterobacteriaceae producing ESBL and AmpC enzymes can be associated with failure of antibiotic therapy and related morbidity and mortality. Their routine detection in microbiology laboratories is still a problem. The aim of this study was to compare the sensitivity of selected phenotypic methods. Material/Methods A total of 106 strains of the Enterobacteriaceae family were tested, in which molecular biology methods confirmed the presence of genes encoding ESBL or AmpC. In ...

  16. Antibacterial activity of ceftriaxone (Ro 13-9904), a beta-lactamase-stable cephalosporin.

    OpenAIRE

    Neu, H C; Meropol, N J; Fu, K P

    1981-01-01

    The in vitro activity of ceftriaxone (Ro 13-9904), a parenteral cephalosporin, was compared with that of other beta-lactam antibiotics. the compound was less active against Staphylococcus aureus and Staphylococcus epidermidis than was cephalothin or cefamandole, but it was comparable to cefoxitin, cefotaxime, and moxalactam in inhibiting most isolates of S. aureus at 3.1 microgram/ml. Ro 13-9904 inhibited Streptococcus pyogenes and Streptococcus pneumoniae at concentrations below 0.25 microgr...

  17. Prevalence of extended-spectrum beta-lactamase-producing bacteria in food

    Directory of Open Access Journals (Sweden)

    Tham J

    2012-10-01

    Full Text Available Johan Tham,1 Mats Walder,2 Eva Melander,2,3 Inga Odenholt11Infectious Diseases Unit, Department of Clinical Sciences, 2Medical Microbiology, Department of Laboratory Medicine, Lund University, Malmö, Sweden; 3Department of Infection Control, Laboratory Medicine, Skåne County, SwedenAbstract: Extended-spectrum β-lactamase (ESBL-producing Enterobacteriaceae with Cefotaximase–München (CTX-M enzymes are rapidly increasing worldwide and pose a threat to health care. ESBLs with CTX-M enzymes have been isolated from animals and different food products, but it is unknown if food imported from the Mediterranean area may be a possible reservoir of these bacteria. During 2007–2008, swab samples from food across different retail outlets (mostly food from the Mediterranean countries and Swedish chicken were collected. Escherichia coli strains from Swedish meat and E. coli isolates from unspecified food from a Swedish food testing laboratory were also examined. In 349 of the 419 swab samples, growth of Enterobacteriaceae was found. In most of the samples, there was also growth of Gram-negative environmental bacteria. Air dry-cured products contained significantly less Enterobacteriaceae isolates compared to lettuces; however, none of the examined Enterobacteriaceae harbored ESBLs. This study did not support the theory that imported food from the Mediterranean area or Swedish domestic food might constitute an important vehicle for the dissemination of ESBL-producing Enterobacteriaceae; however, a spread from food to humans may have occurred after 2008.Keywords: ESBL, antibiotic resistance, zoonosis, food, Enterobacteriaceae

  18. Novel bacteriophage therapy for controlling metallo-beta-lactamase producing Pseudomonas aeruginosa infection in Catfish

    OpenAIRE

    Khairnar, Krishna; Raut, Mahendra P; Rajshree H. Chandekar; Sanmukh, Swapnil G; Waman N. PAUNIKAR

    2013-01-01

    Background The bacteriophage therapy is an effective antimicrobial approach with potentially important applications in medicine and biotechnology which can be seen as an additional string in the bow. Emerging drug resistant bacteria in aquaculture industry due to unrestricted use of antibiotics warrants more sustainable and environmental friendly strategies for controlling fish infections. The isolated bacteria from fish lesions was characterised based on isolation on selective and differenti...

  19. Occurrence of extended-spectrum beta-lactamases (ESBL) in Dutch hospitals

    NARCIS (Netherlands)

    Stobberingh, EE; Arends, J; Hoogkamp-Korstanje, JAA; Goessens, WHF; Visser, MR; Buiting, AGM; Debets-Ossenkopp, YJ; van Ketel, RJ; van Ogtrop, ML; Sabbe, LJM; Voorn, GP; Winter, HLJ; van Zeijl, JH

    1999-01-01

    The prevalence of ESBL was determined among isolates of Escherichia coli (n = 571) and Klebsiella spp. (n = 196) collected during a 1-week study period in 8 university and 3 large regional laboratories all over the Netherlands. 18 isolates were positive for at least one of the screening tests used,

  20. Xenology of beta-lactamases: association of its genetic sources and putative pleiotropism

    OpenAIRE

    Pinheiro, Hugo Manuel Horta, 1979

    2013-01-01

    Em bactérias Gram-negativas, a fase final da síntese do peptidoglicano ocorre no lado periplasmático da membrana celular, envolvendo reacções de carboxipeptidação e transpeptidação mediadas por DD-peptidases membranares. Estas enzimas pertencem a uma família de proteínas conhecidas colectivamente como Penicillin-Binding Proteins (PBPs). As PBPs constituem o alvo preferencial dos antibióticos β-lactâmicos. A molécula do β-lactâmico actua como análogo do substrato enzimático, formando um compl...

  1. SHV-5 extended-spectrum beta-lactamases in clinical isolates of Escherichia coli in Malaysia

    Directory of Open Access Journals (Sweden)

    Subramaniam G

    2006-01-01

    Full Text Available Escherichia coli isolates resistant to ceftazidime isolated in the University Malaya Medical Center (UMMC Kuala Lumpur, Malaysia, between the years 1998 and 2000 were studied for extended-spectrum β -lactamase (ESBL production. All strains were analysed phenotypically and genotypically and found to be ESBL-producing organisms harbouring SHV-5 β-lactamase. This was confirmed by PCR-SSCP and nucleotide sequencing of the blaSHV amplified gene. As there was no evidence of ESBL activity in E. coli prior to this, coupled with the fact that there was a predominance of SHV-5 β-lactamases in Klebsiella pneumoniae isolates in UMMC, we postulate that the E. coli obtained the SHV-5 β-lactamase genes by plasmid transfer from the ESBL-producing K. pneumoniae.

  2. [Extended-spectrum beta-lactamases in enterobacteria other than Escherichia coli and Klebsiella].

    Science.gov (United States)

    Seral García, Cristina; Pardos de la Gándara, María; Castillo García, Francisco Javier

    2010-01-01

    Methods for detecting ESBL-producing Enterobacteriaceae begin by a correct interpretation of the susceptibility profiles, applying the usual criteria for interpretative reading of the antibiogram. Appropriate confirmatory methods will be consequently chosen, based on the inhibition of the enzyme by betalactamases inhibitors, generally clavulanic acid. In case of non-AmpC-producing Enterobacteriaceae, at least two substrates should be used -cefotaxime or ceftriaxone and ceftazidime- to detect enzymes with a low hydrolytic activity against both substrates. Cefepime or AmpC-inhibitors should be recommended for AmpC-producing microorganisms. The identification of the enzymes responsible for the confirmed ESBL phenotype can be performed, either in the clinical laboratory or in reference centres, following a protocol of biochemical and molecular reactions able to detect and characterize, at least, those genes more frequently related to the predominant phenotypic profiles in our region. It is important to know which are the most prevalent combinations enzyme-microorganism, the vehicles for the genetic transmission involved in their dissemination, and the main epidemiological characteristics of the infections that they produce, in order to establish the dimensions of the problem and conduct surveillance studies, with the aim of achieving measures to control the wide spread. PMID:20172418

  3. Occurrence of extended spectrum beta-lactamases among Enterobacteriaceae spp. isolated at a tertiary care institute

    Directory of Open Access Journals (Sweden)

    Kumar M

    2006-01-01

    Full Text Available Increasing resistance to third generation cephalosporins has become a cause for concern especially among Enterobacteriaceae that cause nosocomial infections. The prevalence of extended spectrum β -lactamases (ESBLs among members of Enterobacteriaceae constitutes a serious threat to current β -lactam therapy leading to treatment failure and consequent escalation of costs. A detailed study was initiated to identify the occurrence of ESBLs among the Enterobacteriaceae isolates at a tertiary care hospital using the double disk potentiation technique. Antibiogram profiles were determined to commonly used antibiotics and confirmation of ESBLs production was carried out by the disk diffusion assay using ceftazidime and cefotaxime in the presence and absence of clavulanic acid. Our results indicate that the majority of ESBLs were expressed in Escherichia coli.

  4. Occurrence of Extended-Spectrum and AmpC Beta-Lactamases in Bloodstream Isolates of Klebsiella pneumoniae: Isolates Harbor Plasmid-Mediated FOX-5 and ACT-1 AmpC Beta-Lactamases

    OpenAIRE

    Coudron, Philip E.; Hanson, Nancy D.; Climo, Michael W.

    2003-01-01

    We tested 190 Klebsiella pneumoniae bloodstream isolates recovered from 189 patients in 30 U.S. hospitals in 23 states to determine the occurrence of extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase producers. Based on growth inhibition by clavulanic acid by disk and MIC test methods, 18 (9.5%) of the isolates produced ESBLs. Although the disk diffusion method with standard breakpoints identified 28 cefoxitin-nonsusceptible isolates, only 5 (18%) of these were confirmed as AmpC produ...

  5. Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia

    Science.gov (United States)

    Chishimba, K.; Hang'ombe, B. M.; Muzandu, K.; Mshana, S. E.; Matee, M. I.; Nakajima, C.; Suzuki, Y.

    2016-01-01

    The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum β-lactamase- (ESBL-) producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reaction for detection of blaCTX-M, blaSHV, and blaTEM genes. Overall 20.1%, 77/384, (95% CI; 43.2–65.5%) of total samples analyzed contained ESBL-producing Escherichia coli. The antimicrobial sensitivity test revealed that 85.7% (66/77; CI: 75.7–92) of ESBL-producing E. coli isolates conferred resistance to beta-lactam and other antimicrobial agents. These results indicate that poultry is a potential reservoir for ESBL-producing Escherichia coli. The presence of ESBL-producing Escherichia coli in poultry destined for human consumption requires strengthening of the antibiotic administering policy. This is important as antibiotic administration in food animals is gaining momentum for improved animal productivity in developing countries such as Zambia.

  6. Extended spectrum beta-lactamase-producing Escherichia coli forms filaments as an initial response to cefotaxime treatment

    DEFF Research Database (Denmark)

    Kjeldsen, Thea S. B.; Sommer, Morten Otto Alexander; Olsen, John E.

    2015-01-01

    Background: beta-lactams target the peptidoglycan layer in the bacterial cell wall and most beta-lactam antibiotics cause filamentation in susceptible Gram-negative bacteria at low concentrations. The objective was to determine the initial morphological response of cephalosporin resistant CTX-M-1...

  7. beta-lactamase-producing nontypeable Haemophilus influenzae fails to protect Streptococcus pneumoniae from amoxicillin during experimental acute otitis media

    OpenAIRE

    Westman, E.; Lundin, S.; Hermansson, Ann; Melhus, Åsa

    2004-01-01

    Acute otitis media (AOM) is the most common reason for outpatient antimicrobial therapy. Mixed infections pose a potential problem, since the first-line drug used for the treatment of AOM, amoxicillin, can be neutralized by ß-lactamase-producing pathogens of the upper respiratory tract. To study the effects of a 5-day course of amoxicillin on a mixed middle ear infection, rats were challenged with Streptococcus pneumoniae alone or in combination with ß-lactamase-producing nontypeable Haemophi...

  8. High prevalence of fecal carriage of extended spectrum beta-lactamase/AmpC-producing Enterobacteriaceae in cats and dogs

    OpenAIRE

    Hordijk, J.; Schoormans, A.; Kwakernaak, M.; Duim, B.; Broens, E.; Dierikx, C.M.; Mevius, D.J.; Wagenaar, J. A.

    2013-01-01

    Extended-spectrum-ß-lactamase (ESBL)/AmpC producing Enterobacteriaceae have been reported worldwide amongst isolates obtained from humans, food-producing animals, companion animals, and environmental sources. However, data on prevalence of fecal carriage of ESBL/AmpC producing Enterobacteriaceae in healthy companion animals is limited. This pilot study describes the prevalence of ESBL/AmpC encoding genes in healthy cats and dogs, and cats and dogs with diarrhea. Twenty fecal samples of each g...

  9. Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia

    OpenAIRE

    Chishimba, K.; B.M. Hang’ombe; Muzandu, K.; Mshana, S. E.; Matee, M. I.; Nakajima, C.; Suzuki, Y.

    2016-01-01

    The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum β-lactamase- (ESBL-) producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reac...

  10. EXTENDED SPECTRUM BETA-LACTAMASE PRODUCING E. COLI CONTAMINATION OF CHICKEN MEAT IN THE IRISH RETAIL MARKET

    Directory of Open Access Journals (Sweden)

    Dearbháile Morris

    2014-04-01

    Full Text Available Animals represent potential reservoirs for the dissemination of antimicrobial resistance. Twenty domestically produced chicken meat samples were collected from 19 retail outlets in Ireland, inoculated into Bolton broth and cultured on modified charcoal cefoperazone deoxycholate (mCCDA and Preston agars. Selected representative coliforms included 16 E.coli and 4 Pseudomonas aeruginosa. All E.coli isolates were confirmed as ESBL producers, 15 isolates harbored a blaCTX-M group-1 gene, and none belonged to the E.coli 025b:H4-ST131 clonal group. Pulsed field gel electrophoresis (PFGE analysis identified 13 distinct pulsed field profiles and comparison with more than 300 human clinical isolates of ESBL producing E. coli did not reveal any similarities. ESBL producing E. coli were detected on retail meats in the Irish market place. Although no similarity was apparent between poultry and human isolates this does not preclude a role for ESBL-producing E.coli in meat in dissemination of antimicrobial resistance.

  11. Emergence of Escherichia coli Producing Extended-Spectrum AmpC beta-lactamases (ESAC) in animals

    OpenAIRE

    Jean-YvesMADEC

    2014-01-01

    In both humans and animals, the spread of Extended-Spectrum β-Lactamases (ESBL)/AmpC producers has become a major issue, particularly due to the plasmidic dissemination of most of these genes. Besides, over-expression of the chromosomal ampC gene was largely reported in human and animal Enterobacteriaceae and, more recently, modifications within the coding region of the ampC gene [encoding Extended-spectrum AmpC β-lactamases (ESACs)] were shown to be responsible for an hydrolysis spectrum exp...

  12. Indication of viruses and virus-specific antibodies by ELISA using conjugates based on. beta. -lactamase obtained by genetic engineering

    Energy Technology Data Exchange (ETDEWEB)

    Kharitonenkov, I.G.; Kordym, V.A.; Khristova, M.L.; Leonov, S.V.; Kirillova, V.S.; Chernykh, S.I.

    1987-10-01

    The method of enzyme-linked immunosorbent assay (ELISA), by means of which antigens and antibodies of different origin can be detected with high sensitivity and specificity, is an immunoenzymatic technique based on the use of conjugates, or macromolecular complexes formed by covalent attachment of enzyme molecules to antigen or antibody molecules. Conjugates based on peroxidase, alkaline phosphatase, and beta-galactosidase are most frequently used to construct immunoenzymatic test systems. The use of these enzymes in ELISA, however, is complicated by the fact that they are often present in free or bound form in the biological material under study, and that their substrates either possess low stability, are difficult to synthesize, or are toxic. In this paper, in order to avoid these shortcomings, the authors develop a method for the biosynthesis of lactamase conjugates which is based on genetic engineering, and demonstrate the viability and stability of these conjugates in radioimmunoenzymatic assay of viruses.

  13. Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia.

    Science.gov (United States)

    Chishimba, K; Hang'ombe, B M; Muzandu, K; Mshana, S E; Matee, M I; Nakajima, C; Suzuki, Y

    2016-01-01

    The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum β-lactamase- (ESBL-) producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reaction for detection of bla CTX-M, bla SHV, and bla TEM genes. Overall 20.1%, 77/384, (95% CI; 43.2-65.5%) of total samples analyzed contained ESBL-producing Escherichia coli. The antimicrobial sensitivity test revealed that 85.7% (66/77; CI: 75.7-92) of ESBL-producing E. coli isolates conferred resistance to beta-lactam and other antimicrobial agents. These results indicate that poultry is a potential reservoir for ESBL-producing Escherichia coli. The presence of ESBL-producing Escherichia coli in poultry destined for human consumption requires strengthening of the antibiotic administering policy. This is important as antibiotic administration in food animals is gaining momentum for improved animal productivity in developing countries such as Zambia. PMID:27190518

  14. Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia

    Directory of Open Access Journals (Sweden)

    K. Chishimba

    2016-01-01

    Full Text Available The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum β-lactamase- (ESBL- producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reaction for detection of blaCTX-M, blaSHV, and blaTEM genes. Overall 20.1%, 77/384, (95% CI; 43.2–65.5% of total samples analyzed contained ESBL-producing Escherichia coli. The antimicrobial sensitivity test revealed that 85.7% (66/77; CI: 75.7–92 of ESBL-producing E. coli isolates conferred resistance to beta-lactam and other antimicrobial agents. These results indicate that poultry is a potential reservoir for ESBL-producing Escherichia coli. The presence of ESBL-producing Escherichia coli in poultry destined for human consumption requires strengthening of the antibiotic administering policy. This is important as antibiotic administration in food animals is gaining momentum for improved animal productivity in developing countries such as Zambia.

  15. Amoxicillin concentrations in relation to beta-lactamase activity in sputum during exacerbations of chronic obstructive pulmonary disease

    OpenAIRE

    Brusse-Keizer M; VanderValk P; van der Zanden RW; Nijdam L; van der Palen J; Hendrix R; Movig K

    2015-01-01

    Marjolein Brusse-Keizer,1 Paul VanderValk,2 Rogier W van der Zanden,3 Lars Nijdam,4 Job van der Palen,1,5 Ron Hendrix,6,7 Kris Movig4 1Medical School Twente, 2Department of Pulmonary Medicine, Medisch Spectrum Twente, Enschede, 3Department of Clinical Pharmacy and Toxicology, Maastricht University Medical Centre, Maastricht, 4Department of Clinical Pharmacy, Medisch Spectrum Twente, 5Department of Research Methodology, Measurement and Data Analysis, University of Twente, 6Regional Laboratory ...

  16. Occurrence and sensitivity profile of extended spectrum beta-lactamase-producing Enterobacteriaceae at a tertiary hospital in Southern Brazil

    Directory of Open Access Journals (Sweden)

    Cristina Letícia Rugini

    2015-12-01

    Full Text Available Abstract: INTRODUCTION: Nosocomial infections are closely associated with antimicrobial drug resistance. One of the most important mechanisms of resistance to β-lactam antibiotics is the production of extended spectrum β-lactamases (ESBLs. The objective of the present study was to evaluate the prevalence and antimicrobial susceptibility profile of ESBL-producing strains and to assess the evolution of antimicrobial drug resistance between 2007 and 2013 at the Hospital São Vicente de Paulo, Passo Fundo, State of Rio Grande do Sul, Brazil. METHODS: We conducted a descriptive, observational, cross-sectional study. Bacterial culture was performed from January to December 2013. The antimicrobial susceptibility profile of these cultures was determined using the disk diffusion method. Phenotypic screening for ESBL production was performed using the disk approximation method. RESULTS : We analyzed a total of 19,112 cultures, 11.5% of which were positive for Enterobacteriaceae. Of these, 30.3% of the isolates were positive for ESBL production, and the most prevalent species was Klebsiella sp. (37.5%. Over 95% of these isolates showed reduced susceptibility to all cephalosporins, aztreonam, and amoxicillin/clavulanic acid. The isolates also showed high sensitivity to the following antimicrobials: amikacin, meropenem, and piperacillin/tazobactam. Overall, the resistance rates among ESBL-producing Enterobacteriaceae decreased from 2007 to 2013. CONCLUSIONS : In our hospital, the increased sensitivity to certain antimicrobial agents seems to be directly related to the implementation of improvements in the methods to prevent and control nosocomial infections in addition to the natural development of other resistance mechanisms.

  17. Multidrug-Resistant and Extended Spectrum Beta-Lactamase-Producing Escherichia coli in Dutch Surface Water and Wastewater.

    Directory of Open Access Journals (Sweden)

    Hetty Blaak

    Full Text Available The goal of the current study was to gain insight into the prevalence and concentrations of antimicrobial resistant (AMR Escherichia coli in Dutch surface water, and to explore the role of wastewater as AMR contamination source.The prevalence of AMR E. coli was determined in 113 surface water samples obtained from 30 different water bodies, and in 33 wastewater samples obtained at five health care institutions (HCIs, seven municipal wastewater treatment plants (mWWTPs, and an airport WWTP. Overall, 846 surface water and 313 wastewater E. coli isolates were analysed with respect to susceptibility to eight antimicrobials (representing seven different classes: ampicillin, cefotaxime, tetracycline, ciprofloxacin, streptomycin, sulfamethoxazole, trimethoprim, and chloramphenicol.Among surface water isolates, 26% were resistant to at least one class of antimicrobials, and 11% were multidrug-resistant (MDR. In wastewater, the proportions of AMR/MDR E. coli were 76%/62% at HCIs, 69%/19% at the airport WWTP, and 37%/27% and 31%/20% in mWWTP influents and effluents, respectively. Median concentrations of MDR E. coli were 2.2×10(2, 4.0×10(4, 1.8×10(7, and 4.1×10(7 cfu/l in surface water, WWTP effluents, WWTP influents and HCI wastewater, respectively. The different resistance types occurred with similar frequencies among E. coli from surface water and E. coli from municipal wastewater. By contrast, among E. coli from HCI wastewater, resistance to cefotaxime and resistance to ciprofloxacin were significantly overrepresented compared to E. coli from municipal wastewater and surface water. Most cefotaxime-resistant E. coliisolates produced ESBL. In two of the mWWTP, ESBL-producing variants were detected that were identical with respect to phylogenetic group, sequence type, AMR-profile, and ESBL-genotype to variants from HCI wastewater discharged onto the same sewer and sampled on the same day (A1/ST23/CTX-M-1, B23/ST131/CTX-M-15, D2/ST405/CTX-M-15.In conclusion, our data show that MDR E. coli are omnipresent in Dutch surface water, and indicate that municipal wastewater significantly contributes to this occurrence.

  18. Identifying patients harboring extended-spectrum-beta-lactamase-producing Enterobacteriaceae on hospital admission: derivation and validation of a scoring system.

    Science.gov (United States)

    Tumbarello, Mario; Trecarichi, Enrico Maria; Bassetti, Matteo; De Rosa, Francesco Giuseppe; Spanu, Teresa; Di Meco, Eugenia; Losito, Angela Raffaella; Parisini, Andrea; Pagani, Nicole; Cauda, Roberto

    2011-07-01

    Increases in community-acquired infections caused by extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae have important implications for hospital infection control and empirical antibiotic therapy protocols. We developed and validated a tool for identifying patients harboring these organisms at hospital admission. We retrospectively analyzed chart data for 849 adult inpatients. The derivation cohort included 339 patients admitted to a large hospital in Rome during 2008, with (n = 113) or without (n = 226) culture positivity for ESBL-producing Escherichia coli, Klebsiella spp., or Proteus mirabilis within 48 h after admission. Logistic-regression-based prediction scores were calculated based on variables independently associated with the outcome. The model was validated in a second cohort (n = 510) selected with identical criteria in hospitals in Genoa and Turin during 2009. Prediction scores were based on the following six variables (reported with odds ratio for study outcome and the 95% confidence intervals in brackets): recent (≤ 12 months before admission) hospitalization (5.69 [2.94 to 10.99]), transfer from another health care facility (5.61 [1.65 to 19.08]), Charlson comorbidity score ≥ 4 (3.80 [1.90 to 7.59]), recent (≤ 3 months before admission) β-lactam and/or fluoroquinolone treatment (3.68 [1.96 to 6.91]), recent urinary catheterization (3.52 [1.96 to 6.91]), and age ≥ 70 years (3.20 [1.79 to 5.70]). The model displayed good calibration and good-to-excellent discrimination in the derivation and validation sets (Hosmer-Lemshow χ(2) = 15.28 and 14.07; P = 0.17 and 0.23; areas under the receiver-operating characteristic curve, 0.83 and 0.92). It reliably identified patients likely to be harboring ESBL-producing Enterobacteriaceae at hospital admission who may need special infection control measures. Further study is needed to confirm this model's potential as a guide for prescribing empirical antibiotic therapy. PMID:21537020

  19. Pseudomonas aeruginosa biofilms exposed to imipenem exhibit changes in global gene expression and beta-lactamase and alginate production

    DEFF Research Database (Denmark)

    Bagge, Niels; Schuster, Martin; Hentzer, Morten;

    2004-01-01

    , on gene expression in biofilm populations. Many genes showed small but statistically significant differential expression in response to imipenem. We identified 34 genes that were induced or repressed in biofilms exposed to imipenem more than fivefold compared to the levels of induction or repression...

  20. Epidemiology and virulence of VIM-4 metallo-beta-lactamase-producing Pseudomonas aeruginosa isolated from burn patients in eastern Algeria.

    Science.gov (United States)

    Meradji, Samah; Barguigua, Abouddihaj; Bentakouk, Mohamed Cherif; Nayme, Kaotar; Zerouali, Khalid; Mazouz, Dekhil; Chettibi, Houria; Timinouni, Mohammed

    2016-06-01

    In this study, we investigated the prevalence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in burn patients from eastern Algeria, CRPA virulence factors and the molecular epidemiology of CRPA. The overall prevalence of CRPA was 48.38%. Seven (46.66%) isolates were metallo-β-lactamases (MBL) producers and contained the MBL genes blaVIM-4 (n=6) and blaVIM-2 (n=1). Risk factors for CRPA infection were urinary catheter use and intubation (p=0.008). A high percentage of virulence factors (86.6% of these isolates were able to produce protease; 73.3% of isolates has DNase; and 66.6% were haemolysin positive) was observed in CRPA isolates. Among the seven MBL-producing isolates, four had the same clonal profile. The class 1 integrons, which contained the aadA7 gene cassette, were detected in six isolates. The 16SrRNA methylase gene, rmtB, was detected in one strain. All CRPA isolates were biofilm formers. A study on the kinetics of biofilm production revealed that biofilm production increased when the concentration of imipenem or ciprofloxacin and the incubation time increased. This is the first study to report the presence of VIM-4-producing P. aeruginosa from North Africa and also of the high prevalence of CRPA isolates. Based on our study of burn unit patients, the high percentage of P. aeruginosa with virulence factors and multi-drug resistance is alarming. PMID:27156788

  1. Antimicrobial Resistance Pattern and Their Beta-Lactamase Encoding Genes among Pseudomonas aeruginosa Strains Isolated from Cancer Patients

    Directory of Open Access Journals (Sweden)

    Mai M. Zafer

    2014-01-01

    Full Text Available This study was designed to investigate the prevalence of metallo-β-lactamases (MBL and extended-spectrum β-lactamases (ESBL in P. aeruginosa isolates collected from two different hospitals in Cairo, Egypt. Antibiotic susceptibility testing and phenotypic screening for ESBLs and MBLs were performed on 122 P. aeruginosa isolates collected in the period from January 2011 to March 2012. MICs were determined. ESBLs and MBLs genes were sought by PCR. The resistant rate to imipenem was 39.34%. The resistance rates for P. aeruginosa to cefuroxime, cefoperazone, ceftazidime, aztreonam, and piperacillin/tazobactam were 87.7%, 80.3%, 60.6%, 45.1%, and 25.4%, respectively. Out of 122 P. aeruginosa, 27% and 7.4% were MBL and ESBL, respectively. The prevalence of blaVIM-2, blaOXA-10-, blaVEB-1, blaNDM-, and blaIMP-1-like genes were found in 58.3%, 41.7%, 10.4%, 4.2%, and 2.1%, respectively. GIM-, SPM-, SIM-, and OXA-2-like genes were not detected in this study. OXA-10-like gene was concomitant with VIM-2 and/or VEB. Twelve isolates harbored both OXA-10 and VIM-2; two isolates carried both OXA-10 and VEB. Only one strain contained OXA-10, VIM-2, and VEB. In conclusion, blaVIM-2- and blaOXA-10-like genes were the most prevalent genes in P. aeruginosa in Egypt. To our knowledge, this is the first report of blaVIM-2, blaIMP-1, blaNDM, and blaOXA-10 in P. aeruginosa in Egypt.

  2. High prevalence of fecal carriage of extended spectrum beta-lactamase/AmpC-producing Enterobacteriaceae in cats and dogs

    NARCIS (Netherlands)

    Hordijk, J.; Schoormans, A.; Kwakernaak, M.; Duim, B.; Broens, E.; Dierikx, C.M.; Mevius, D.J.; Wagenaar, J.A.

    2013-01-01

    Extended-spectrum-ß-lactamase (ESBL)/AmpC producing Enterobacteriaceae have been reported worldwide amongst isolates obtained from humans, food-producing animals, companion animals, and environmental sources. However, data on prevalence of fecal carriage of ESBL/AmpC producing Enterobacteriaceae in

  3. Frequency and Antimicrobial Resistance Pattern of Extended Spectrum Beta Lactamases Producing Escherichia coli Strains Isolated from Urinary Infections

    Directory of Open Access Journals (Sweden)

    Mohammad Rasoul Tohidnia

    2013-07-01

    Full Text Available AbstractBackground and objectives: Escherichia coli is the most common cause of urinary tract infections. The resistance of Escherichia coli strains to broad spectrum cephalosporins may be mediated by extended spectrum β-lactamases (ESBLs has now raised in worldwide. Frequency and spectrum awareness of drug resistance in these isolates will be useful in choosing more effective methods of control and treatment. Materials and methods: This cross - sectional study was done on 100 Uropathogenic E. coli strains collected from selected center and then were identified by biochemical tests. All of the samples were screened by DAD method. The phenotypes of ESBL were determined using the disk diffusion synergy test(DDST as recommended by the interpretative guidelines of the NCCLS. SPSS16 was used for statistical analysis. Results: A total of one hundred isolates,89(89%were found to be resistant at least on of the indicators cephalosporin tested according to NCCLS guideline.55(55%of the isolates were resistant to all the indicators tested. Sixty tree isolates (63% were ESBL producers. The E. coli strains showed high susceptibility to imipenem (66/7%and amikacin (65/1%.Conclusion: Our results show that the most appropriate antibiotic to be used for empirical therapy are imipenem and amikacin.High prevalence of ESBL in our hospital cannot be ignored. ESBL producers can be detected by DDST and phenotypic confirmatory test with equal efficacy.we recommend DDSTusing multiple antibiotics in all microbiology units as a routine screening test.

  4. Rectal carriage of extended-spectrum beta-lactamase-producing gram-negative bacilli in community settings in Madagascar.

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    Perlinot Herindrainy

    Full Text Available BACKGROUND: Extended-spectrum ß-lactamase-producing Enterobacteria (ESBL-PE emerged at the end of the 1980s, causing nosocomial outbreaks and/or hyperendemic situations in hospitals and long-term care facilities. In recent years, community-acquired infections due to ESBL-PE have spread worldwide, especially across developing countries including Madagascar. OBJECTIVES: This study aimed to determine the prevalence and risk factors of intestinal carriage of ESBL-PE in the community of Antananarivo. METHODS: Non-hospitalized patients were recruited in three health centers in different socio economic settings. Fresh stool collected were immediately plated on Drigalski agar containing 3 mg/liter of ceftriaxone. Gram-negative bacilli species were identified and ESBL production was tested by a double disk diffusion (cefotaxime and ceftazidime +/- clavulanate assay. Characterization of ESBLs were perfomed by PCR and direct sequencing. Molecular epidemiology was analysed by Rep-PCR and ERIC-PCR. RESULTS: 484 patients were screened (sex ratio  =  1.03, median age 28 years. 53 ESBL-PE were isolated from 49 patients (carrier rate 10.1%. The isolates included Escherichia coli (31, Klebsiella pneumoniae (14, Enterobacter cloacae (3, Citrobacter freundii (3, Kluyvera spp. (1 and Pantoae sp. (1. In multivariate analysis, only the socioeconomic status of the head of household was independently associated with ESBL-PE carriage, poverty being the predominant risk factor. CONCLUSIONS: The prevalence of carriage of ESBL in the community of Antananarivo is one of the highest reported worldwide. This alarming spread of resistance genes should be stopped urgently by improving hygiene and streamlining the distribution and consumption of antibiotics.

  5. Dynamics of spread of intestinal colonization with extended-spectrum beta-lactamases in E.coli: a mathematical model

    DEFF Research Database (Denmark)

    Philipsen, Kirsten Riber; Bootsma, M. C. J.; Leverstein-van Hall, M.A.; Cohen Stuart, J.; Bonten, M.J.M.

    parameters. The patient flow data was studied by survival analysis. This enabled us to get an estimate for the time to readmission when discharged from either the low-risk or high-risk hospital wards. It is hypothesized that readmission plays a role for the spread of resistant bacteria. The high prevalence...... importance of readmission and the effect of different interventions on the prevalence. There are several theories with regards to the spread of ESBL resistant bacteria, but the actual prevalence data is very sparse. Based on the available data we have developed an adequate model that can explain the increase...... in prevalence from year 2000. It has not been possible to separate the e®ect from conjugation and cross-transfer on the ESBL prevalence of type CTX-M, as several combination of the cross-transfer rate and conjugation rate give a good ¯t to data. However, the model predicts that a minimum of 57% of...

  6. “UROPATHOGENS”: PREVALENCE AND ANTIBIOGRAM OF GRAM NEGATIVE BACILLI WITH SPECIAL REFERENCE TO EXTEN DED SPECTRUM BETA LACTAMASE PRODUCTION

    Directory of Open Access Journals (Sweden)

    Vandana

    2012-12-01

    Full Text Available ABSTRACT: This study was performed on culture and sensitivit y of 6,951 urine samples, received in the Department of Microbiology, Christia n Medical College & Hospital, Ludhiana from out patients and in patients having urinary tra ct infection (UTI .A total of 2,276 samples were found out to be culture positive, out of which 1,727 samples yielded gram negative organisms. Various isolates included 1,237 Escheric hia coli (E. coli, 262 Klebsiella pneumoniae,47 Acinetobacter lwoffi, 39 Proteus mirab ilis,39 Enterobacter aerogenes and 03 Pseudomonas aeruginosa. Extended spectrum beta lacta mase (ESBL production was studied in multidrug resistant E. coli. And Klebsiella pneumoni ae, out of which 28.29% E.coli and 30.53% Klebsiella pneumoniae yielded positive results. Our r esults suggest that the physician should be aware of high prevalence of ESBL producing E.coli and Klebsiella pneumonia which are the two common uropathogens, and should plan their therapy re gime accordingly. However, Acinetobacter species were mainly associate d with nosocomial UTI whereas Enterobacter species were isolated mostly from out pa tients. Various uropathogens causing community acquired as well as nosocomial UTI showed poor response to cephalexin whereas resistant strains from both types of UTI exhibited g ood susceptibility to piperacillin/tazobactum combination.

  7. Detection of AmpC Beta-lactamases among Escherichia coli isolates at a tertiary care hospital in Karnataka

    OpenAIRE

    Smitha O. Bagali; B.V. Peerapur

    2013-01-01

    Background & objective: AmpC β-lactamases are clinically significant because they may confer resistance to a wide variety of β-lactam drugs, including α-methoxy-β-lactams, such as cefoxitin, narrow-, expanded- and broad-spectrum cephalosporins, β-lactam-β-lactamase inhibitor combinations and aztreonam. Although reported with increasing frequency the true occurrence in different organisms remains unknown. The present study was conducted to determine the occurrence of AmpC β-lactamases among th...

  8. Prevalence of AmpC and other beta-lactamases in enterobacteria at a large urban university hospital in Brazil

    OpenAIRE

    Dias, Rubens Clayton da Silva; Borges-Neto, Armando Alves; Ferraiuoli, Giovanna Ianini D’Almeida; de-Oliveira, Márcia P.; Riley, Lee W.; Moreira, Beatriz Meurer

    2007-01-01

    Production of extended-spectrum β-lactamases (ESBL) has been reported in virtually all species of Enterobacteriaceae, which greatly complicates the therapy of infections caused by these organisms. However, the frequency of isolates producing AmpC β-lactamases, especially plasmid mediated AmpC (pAmpC), is largely unknown. These β-lactamases confer resistance to extended spectrum cephalosporins and aztreonam, a multidrug-resistant (MDR) profile. The aim of the present study was to determine the...

  9. Detection of AmpC beta-lactamase in Escherichia coli: comparison of three phenotypic confirmation assays and genetic analysis

    OpenAIRE

    Peter-Getzlaff, S.; Polsfuss, S; Poledica, M.; Hombach, M; Giger, J; Böttger, E.C.; R. Zbinden; Bloemberg, G V

    2011-01-01

    Two mechanisms account for AmpC activity in Escherichia coli, namely, mutations in the ampC promoter and attenuator regions resulting in ampC overexpression and acquisition of plasmid-carried ampC genes. In this study, we analyzed 51 clinical E. coli isolates with reduced susceptibility to amoxicillin-clavulanic acid, piperacillin-tazobactam, or extended-spectrum cephalosporins for the presence of AmpC production. Three phenotypic AmpC confirmation assays (cefoxitin-cloxacillin disk diffusion...

  10. Evaluation of Methods for AmpC Beta-Lactamase in Gram Negative Clinical Isolates from Tertiary Care Hospitals

    OpenAIRE

    Singhal S; Mathur T; Khan S; Upadhyay D; Chugh S; Gaind R; Rattan A

    2005-01-01

    The purpose of this study was to simultaneously screen for Extended-spectrum b-lactamases (ESBL) and AmpC b-lactamases in gram negative clinical isolates from four tertiary care hospitals and further to compare two detection methods three-dimensional extraction method and AmpC disk test for AmpC b-lactamases. A total of 272 isolates were screened for ESBL and AmpC β-lactamase by modified double disk approximation method (MDDM). Synergy observed between disks of ceftazidime/cefotaxime a...

  11. [Extended spectrum beta lactamases (ESBL) production in Acinetobacter baumannii strains isolated from Chilean hospitals belonging to VIII Region].

    Science.gov (United States)

    Pino I, Carolina; Domínguez Y, Mariana; González R, Gerardo; Bello T, Helia; Sepúlveda A, Marcela; Mella M, Sergio; Zemelman M, Claudia; Zemelman Z, Raúl

    2007-04-01

    The resistance of Acinetobacter baumannii to ss-lactam antibiotics is mainly due to the synthesis of ss-lactamases. From a clinical point of view, this bacteria and others, grouped under the acronym SPACE (S: Serratia, P: Pseudomonas, A: Acinetobacter, C: Citrobacter, E: Enterobacter) are essentially Amp-C ss-lactamases producers. There is no local information about ESBL presence in Acinetobacter. We studied ESBL production using the Ho and col. technique modified by adding cloxacillin as chromosomal ss-lactamases inhibitor. From 69 isolates, with resistance to at least one third generation cephalosporin, only 7 showed positive synergy test. Four of these amplified for TEM family gene, and one of these amplified also for the OXA family. Our study found a low ESBL production percentage, which agrees with the premise of Amp-C as the main mechanism of resistance to ss-lactam antibiotics in A. baumannii. However, the ESBL description in these bacteria emphasizes the capacity of expressing multiple resistance mechanisms. PMID:17453072

  12. Detection of Extended-Spectrum Beta-Lactamases among Enterobacteriaceae by Use of Semiautomated Microbiology Systems and Manual Detection Procedures▿

    OpenAIRE

    Wiegand, Irith; Geiss, Heinrich K.; Mack, Dietrich; Stürenburg, Enno; Seifert, Harald

    2007-01-01

    Three commercially available microbiology identification and susceptibility testing systems were compared with regard to their ability to detect extended-spectrum β-lactamase (ESBL) production in Enterobacteriaceae, i.e., the Phoenix Automated Microbiology System (BD Diagnostic Systems, Sparks, MD), the VITEK 2 System (bioMérieux, Marcy l'Etoile, France), and the MicroScan WalkAway-96 System (Dade Behring, Inc., West Sacramento, CA), using routine testing panels. One hundred fifty putative ES...

  13. Comparable high rates of extended-spectrum-beta-lactamase-producing Escherichia coli in birds of prey from Germany and Mongolia.

    Directory of Open Access Journals (Sweden)

    Sebastian Guenther

    Full Text Available Frequent contact with human waste and liquid manure from intensive livestock breeding, and the increased loads of antibiotic-resistant bacteria that result, are believed to be responsible for the high carriage rates of ESBL-producing E. coli found in birds of prey (raptors in Central Europe. To test this hypothesis against the influence of avian migration, we initiated a comparative analysis of faecal samples from wild birds found in Saxony-Anhalt in Germany and the Gobi-Desert in Mongolia, regions of dissimilar human and livestock population characteristics and agricultural practices. We sampled a total of 281 wild birds, mostly raptors with primarily north-to-south migration routes. We determined antimicrobial resistance, focusing on ESBL production, and unravelled the phylogenetic and clonal relatedness of identified ESBL-producing E. coli isolates using multi-locus sequence typing (MLST and macrorestriction analyses. Surprisingly, the overall carriage rates (approximately 5% and the proportion of ESBL-producers among E. coli (Germany: 13.8%, Mongolia: 10.8% were similar in both regions. Whereas bla(CTX-M-1 predominated among German isolates (100%, bla(CTX-M-9 was the most prevalent in Mongolian isolates (75%. We identified sequence types (STs that are well known in human and veterinary clinical ESBL-producing E. coli (ST12, ST117, ST167, ST648 and observed clonal relatedness between a Mongolian avian ESBL-E. coli (ST167 and a clinical isolate of the same ST that originated in a hospitalised patient in Europe. Our data suggest the influence of avian migratory species in the transmission of ESBL-producing E. coli and challenge the prevailing assumption that reducing human influence alone invariably leads to lower rates of antimicrobial resistance.

  14. Pseudomonas aeruginosa biofilms exposed to imipenem exhibit changes in global gene expression and beta-lactamase and alginate production

    DEFF Research Database (Denmark)

    Bagge, N.; Schuster, M.; Hentzer, Morten;

    2004-01-01

    The lungs of cystic fibrosis (CF) patients are commonly colonized with Pseudomonas aeruginosa biofilms. Chronic endobronchial P. aeruginosa infections are impossible to eradicate with antibiotics, but intensive suppressive antibiotic therapy is essential to maintain the lung function of CF patients....... The treatment often includes beta-lactam antibiotics. How these antibiotics influence gene expression in the surviving biofilm population of P. aeruginosa is not clear. Thus, we used the microarray technology to study the effects of subinhibitory concentrations of a beta-lactam antibiotic, imipenem......, on gene expression in biofilm populations. Many genes showed small but statistically significant differential expression in response to imipenem. We identified 34 genes that were induced or repressed in biofilms exposed to imipenem more than fivefold compared to the levels of induction or repression...

  15. [Epidemiologic study on the prevalence of Enterobacter, Serratia and Pseudomonas strains, producers of cefoxitin-inducible beta-lactamases].

    Science.gov (United States)

    Marone, P; Concia, E; Perversi, L; Cruciani, M

    1986-01-01

    Resistance of Enterobacter, Serratia and pseudomonas strains to newer cephalosporins is often associated with stable derepression of synthesis of the chromosomal betalactamases. Similar resistance is developed by enzyme inducible strains in response to betalactamases inducers. This finding poses many clinical problems including emergence of resistance during therapy with the drugs. In this study we evaluated the MICs of several new betalactam compounds against 76 Enterobacter, Serratia and Pseudomonas strains before and after cefoxitin-induction of betalactamases. The MICs against several Enterobacter strains (45%) after cefoxitin induction were elevated four fold or more. Serratia strains showed no significant variations of the MICs after cefoxitin induction. The MICs of piperacillin against many Pseudomonas strains (78%) after cefoxitin induction were elevated four fold or more. These data were confirmed using cefoxitin disk approximation test. Outbreaks of nosocominal infection with these multiresistant bacteria and spread of the strains throughout the hospital are already being seen. Control of these problems can only be achieved through the judicious and restricted use of these new antibiotics. PMID:3103651

  16. Antibiotic Resistance Pattern and the Prevalence of Extended Spectrum Beta-Lactamases (ESBLs in Urinary Isolates of Klebsiella Pneumoniae

    Directory of Open Access Journals (Sweden)

    Shahraki, SH. (PhD

    2014-11-01

    Full Text Available Background and Objective: Klebsiella pneumoniae is an opportunistic nosocomial pathogen causing a variety of infections including urinary tract infections, pneumonia, septicemia, wound infections and infections in the intensive care units. Since the ESBL producing Klebsiella pneumoniae strains are increasingly causing urinary tract infections, we aim to assess antibiotic resistance pattern and evaluate the prevalence of ESBL in Klebsiella pneumoniae isolated from urinary tract infections. Material and Methods: this cross-sectional study was conducted on 122 Klebsiella pneumoniae strains collected from Zahedan hospitals. After final identification of isolates, antibiotic susceptibility tests were carried out by using disk diffusion in agar method for 16 antibiotics and ESBL production was determined by the combined disk method. Results: The Klebsiella pneumoniae strains showed susceptibility to imipenem and amikacin ( 94.3% ,chloramphenicol (88.5% , gentamicin (81.1% , ciprofloxacin (80.3% , cefepime (73% ,streptomycin (72.1%, nalidixic acid (68% , tetracycline (65.6%, and cefotaxime, ceftazidime, cefpodoxime (62.3% . The resistance of strains was seen to nitrofurantoin (53.3%, cotrimoxazole (39.3%, Cefpodoxime (37.7%, cefotaxime (36.9%, ceftriaxone (36.1%, aztreonam (34.4%, ceftazidime (32.8%. Thirty-eight isolates (31.1% were shown to produce ESBLs. Conclusion: A high rate of resistance was observed to most of the antibiotics among ESBL producing strains; therefore, it is important to be careful about the use of antibiotics and identification of ESBL using phenotypic methods.

  17. Extended-spectrum beta-lactamases in ceftazidime-resistant Escherichia coli and Klebsiella pneumoniae isolates in Turkish hospitals

    Directory of Open Access Journals (Sweden)

    Hosoglu S

    2007-01-01

    Full Text Available Purpose: To study the prevalence of TEM-, SHV- and GES-type β -lactamases among Escherichia coli and Klebsiella pneumoniae strains having ceftazidime MICs higher than 2 mg/L. Methods: A total of 63 E. coli and 41 K. pneumoniae isolated from five different university hospitals were studied for the existence of TEM-, SHV- and GES-type β -lactamases. Susceptibility tests were carried out according to the criteria of National Committee for Clinical Laboratory Standards. MICs were obtained by agar dilution method. Existence of extended-spectrum β -lactamases (ESBLs were assessed by double-disc synergy test (DDST. Existence of the above-mentioned β -lactamase genes were studied both by PCR with specific oligonucleotide primers and isoelectric focusing methods. Results: None of the isolates were carbapenem-resistant. DDSTs were positive in 50 (79.3% and 33 (80.5% of E. coli and K. pneumoniae , respectively. TEM gene was detected in 41 (65.1% and 19 (46.3%, whereas SHV gene in 18 (28.6% and 20 (48.8% of E. coli and K. pneumoniae strains, respectively. GES genes were not detected. Conclusions: TEM and SHV genes are highly prevalent among ESBL-producing E. coli and K. pneumoniae , whereas GES-type ESBLs are absent and found not to be responsible of ceftazidime resistance in Turkish hospitals.

  18. Detection of AmpC Beta-lactamases among Escherichia coli isolates at a tertiary care hospital in Karnataka

    Directory of Open Access Journals (Sweden)

    Smitha O. Bagali

    2013-01-01

    Full Text Available Background & objective: AmpC β-lactamases are clinically significant because they may confer resistance to a wide variety of β-lactam drugs, including α-methoxy-β-lactams, such as cefoxitin, narrow-, expanded- and broad-spectrum cephalosporins, β-lactam-β-lactamase inhibitor combinations and aztreonam. Although reported with increasing frequency the true occurrence in different organisms remains unknown. The present study was conducted to determine the occurrence of AmpC β-lactamases among the clinical isolates of Escherichia coli. Methods: A total of 100 non-repeat clinical isolates obtained from urine, pus, sputum, blood and body fluids were taken. All the isolates were screened for AmpC β-lactamases by standard disc diffusion breakpoint for cefoxitin (30µg. Isolates with zone diameter less than 18 mm were tested for AmpC activity by AmpC disc test. Results: Of the 100 isolates that were tested, 30 yielded cefoxitin zone diameters less than 18 mm (screen positive. Production of AmpC β-lactamase was detected in 24 isolates by AmpC disc test. Conclusion: AmpC disc test can be used as a simple, convenient and rapid screening test for detection of AmpC β lactamase in clinical laboratories.

  19. The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing

    International Nuclear Information System (INIS)

    The spreading of extended-spectrum β-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and blaTEM+CTx-M was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes. - Yangtze River is heavily polluted by ESBL-producing TC bacteria and Class 1 integrons play an important role in multi-drug resistance.

  20. The influence of allotypes on the IgG subclass response to chromosomal beta-lactamase of Pseudomonas aeruginosa in cystic fibrosis patients

    DEFF Research Database (Denmark)

    Ciofu, O; Pressler, T; Pandey, J P; Høiby, N

    1997-01-01

    longitudinal study. Increased levels of IgG2 were associated with the presence of GM 23 allotype. IgG3 a beta ab levels were the lowest for subjects with the GM 1,2,3,17 23 5,21 and GM 1,3,17 21 phenotypes and the highest in subjects with GM 3,23,5 and GM 3,5. No significant differences in IgG1 and IgG4 a beta......, lower IgG4 a beta ab levels and poorer lung function than patients with G3M*21 G1M*1/ G3M*21 G1M*1 genotype. An influence of the allotypes on the clinical course of chronic lung infection with Ps. aeruginosa in patients with CF is suggested....

  1. Molecular epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli in Tunisia and characterization of their virulence factors and plasmid addiction systems

    OpenAIRE

    Mnif, Basma; Harhour, Hela; Jdidi, Jihène; Mahjoubi, Faouzia; Genel, Nathalie; Arlet, Guillaume; Hammami, Adnene

    2013-01-01

    Background Extended-spectrum β-lactamases (ESBLs), particularly CTX-M- type ESBLs, are among the most important resistance determinants spreading worldwide in Enterobacteriaceae. The aim of this study was to characterize a collection of 163 ESBL-producing Escherichia coli collected in Tunisia, their ESBL-encoding plasmids and plasmid associated addiction systems. Results The collection comprised 163 ESBL producers collected from two university hospitals of Sfax between 1989 and 2009. 118 isol...

  2. Carriage of Extended-Spectrum Beta-Lactamase-Plasmids Does Not Reduce Fitness but Enhances Virulence in Some Strains of Pandemic E. coli Lineages

    OpenAIRE

    Schaufler, Katharina; Semmler, Torsten; Pickard, Derek J.; de Toro, María; de la Cruz, Fernando; Lothar H Wieler; Ewers, Christa; Guenther, Sebastian

    2016-01-01

    Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL...

  3. Comparative possession of Shiga toxin, intimin, enterohaemolysin and major extended spectrum beta lactamase (ESBL) genes in Escherichia coli isolated from backyard and farmed poultry

    OpenAIRE

    Samanta, I.; Joardar, S. N.; Das, P. K.; Sar, T. K.

    2015-01-01

    The present work was conducted to compare the occurrence of Escherichia coli possessing virulence and ESBL genes in backyard and farmed poultry. Three hundred and sixty samples from the poultry kept in backyard system and 120 samples from the farmed birds were collected from West Bengal, India. Among the E. coli isolates of backyard poultry (O2, O10, O25, O55, O60, O106, UT), none of them possessed any of the Shiga toxin genes and eight E. coli isolates (8/272; 2.9%) harboured eaeA gene alone...

  4. Frequency and Sensitivity of Extended Spectrum Beta-Lactamase Positive Organisms in a Secondary and Tertiary Level Hospital Network in Dhaka

    Directory of Open Access Journals (Sweden)

    Shah Md Zahurul Haque Asna

    2015-05-01

    Full Text Available Background: Extended spectrum β-lactamase (ESBL positive organisms are now a global health concern including in Bangladesh. These are associated with treatment failure, increased morbidity and mortality and increased health care costs. In this study, frequency of ESBL positive organisms in some health care centres in Dhaka city has been observed and their current status of antibiogram has also been observed. Objective: To observe the current status of antibiogram of ESBL positive organisms. Materials and Methods: This cross-sectional study was done in the Department of Microbiology, Bangladesh Institute of Health Sciences (BIHS General Hospital, Dhaka, Bangladesh from March, 2012 to February, 2013. Only E. coli and Klebsiella spp. from pus and urine specimens were included in this study. Isolation, identification and antibiotic sensitivity of the organisms were done by standard procedures. Results: Organisms (Escherichia coli and Klebsiella spp. isolated from urine and pus collected from different sites of 472 subjects were studied. Predominant organisms were Escherichia coli (82.8% and remaining 17.2% were Klebsiella spp. ESBL positive organisms were higher in Escherichia coli (54.5% than in Klebsiella spp. (44.4% and higher in pus (77.0% than in urine (49.1% isolates. Imipenem is the most effective drug for treating ESBL positive organisms followed by colistin, tigecycline and piperacillin/tazobactam. Conclusion: Imipenem, colistin, tigecycline and piperacillin/tazobactam drugs should be kept reserved and used only when other effective drugs are not available so that emergence of resistance against these drugs is deferred. While reporting the culture and sensitivity tests, the ESBL positive organisms should be pointed out with comment like this – “The organisms are ESBL positive and resistant to penicillins, cephalosporins and monobactams”.

  5. Crystallographic Studies of Two Bacterial AntibioticResistance Enzymes: Aminoglycoside Phosphotransferase (2')-Ic and GES-1\\beta-lactamase

    Energy Technology Data Exchange (ETDEWEB)

    Brynes, Laura; /Rensselaer Poly.

    2007-10-31

    Guiana Extended-Spectrum-1 (GES-1) and Aminoglycoside phosphotransferase (2')-Ic (APH(2')-Ic) are two bacteria-produced enzymes that essentially perform the same task: they provide resistance to an array of antibiotics. Both enzymes are part of a growing resistance problem in the medical world. In order to overcome the ever-growing arsenal of antibiotic-resistance enzymes, it is necessary to understand the molecular basis of their action. Accurate structures of these proteins have become an invaluable tool to do this. Using protein crystallography techniques and X-ray diffraction, the protein structure of GES-1 bound to imipenem (an inhibitor) has been solved. Also, APH(2')-Ic has been successfully crystallized, but its structure was unable to be solved using molecular replacement using APH(2')-Ib as a search model. The structure of GES-1, with bound imipenem was solved to a resolution of 1.89A, and though the inhibitor is bound with only moderate occupancy, the structure shows crucial interactions inside the active site that render the enzyme unable to complete the hydrolysis of the {beta}-lactam ring. The APH(2')-Ic dataset could not be matched to the model, APH(2')-Ib, with which it shares 25% sequence identity. The structural information gained from GES-1, and future studies using isomorphous replacement to solve the APH(2')-Ic structure can aid directly to the creation of novel drugs to combat both of these classes of resistance enzymes.

  6. Pseudomonas aeruginosa chromosomal beta-lactamase in patients with cystic fibrosis and chronic lung infection. Mechanism of antibiotic resistance and target of the humoral immune response

    DEFF Research Database (Denmark)

    Ciofu, Oana

    2003-01-01

    The intensive antibiotic treatment of cystic fibrosis (CF) patients with chronic lung infection with Pseudomonas aeruginosa has improved the survival rate and the clinical condition of Danish patients. Acquirement of resistance to anti-pseudomonal antibiotics is one of the main drawbacks of this...... therapeutic strategy and our results showed the development of resistance of P. aeruginosa to several antibiotics during 25 years of intensive antibiotic treatment. Our studies have been concentrating on the development of resistance to beta-lactam antibiotics. We have shown an association between the...... induction to even higher levels during treatment with beta-lactam antibiotics, were the most frequent phenotype found among resistant Danish P. aeruginosa CF isolates. We have also shown that the high alginate producing P. aeruginosa isolates, that characterize the chronic lung infection in CF patients, are...

  7. Investigation of the presence of extended spectrum beta-lactamases (ESBL) in multiresistant strains of E. Coli and salmonella species originated from domestic animals

    OpenAIRE

    Filipović Irina; Mišić D.; Ašanin Ružica

    2007-01-01

    Bacterial strains which possess genes to produce ESBL most often are multiresistant and also carry genes responsible for the resistance to most other antibiotics, including aminoglycosides, sulfamethoxazole+trimethoprim and fluoroquinolones. Therefore, practically the biggest contemporary clinical problem are infections of humans and animals caused by ESBL-producing strains of E. coli, Kleibsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella and Shigella species. The investigatio...

  8. Beta-lactam induction of ISEcp1B-mediated mobilization of the naturally occurring bla(CTX-M) beta-lactamase gene of Kluyvera ascorbata

    OpenAIRE

    Nordmann, Patrice; Lartigue, Marie-Frédérique; Poirel, Laurent

    2008-01-01

    ISEcp1B is an insertion element associated with the emerging expanded-spectrum β-lactamase blaCTX-M genes in Enterobacteriaceae. Because ISEcp1B-blaCTX-M positive strains may be identified from humans and animals, the ability of this insertion sequence to mobilize the blaCTX-M-2 gene was tested from its progenitor Kluyvera ascorbata to study the effects of amoxicillin/clavulanic and cefquinome as enhancers of transposition. These β-lactam molecules are administered parenterally to treat infec...

  9. Carriage of Extended-Spectrum Beta-Lactamase-Plasmids Does Not Reduce Fitness but Enhances Virulence in Some Strains of Pandemic E. coli Lineages.

    Science.gov (United States)

    Schaufler, Katharina; Semmler, Torsten; Pickard, Derek J; de Toro, María; de la Cruz, Fernando; Wieler, Lothar H; Ewers, Christa; Guenther, Sebastian

    2016-01-01

    Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-"cured" variants (PCV), and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM) assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed toward similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages. PMID:27014251

  10. Prevalence of Extended-Spectrum Beta-Lactamase-producing Enterobacteriaceae in humans living in municipalities with high and low broiler density

    OpenAIRE

    Huijbers, P.M.C.; Kraker, de, A Bram; Graat, E.A.M.; Hoek, van, E.; Santen, M.G.M.; Jong, de, D.; Duijkeren, van, E.; Greeff, de, S.C.

    2013-01-01

    Prevalence of, and risk factors for, carriage of extended-spectrum ß-lactamase (ESBL) -producing Enterobacteriaceae were determined for 1025 Dutch adults in municipalities with either high or low broiler densities. Overall prevalence of ESBL carriage was 5.1%. The hypothesis that individuals in areas with high broiler densities are at greater risk for ESBL carriage was rejected, as the risk was lower (OR = 0.45; p 0.009) for these individuals. Owning a horse increased the risk (OR = 4.69; p =...

  11. Phenotypic and Molecular Characterisation of Extended-Spectrum Beta-Lactamase Producing Escherichia coli Obtained from Animal Fecal Samples in Ado Ekiti, Nigeria

    OpenAIRE

    Olugbenga Adekunle Olowe; Olufunmilayo Adewumi; Gbolabo Odewale; Olusola Ojurongbe; Olusolabomi Jose Adefioye

    2015-01-01

    Production of extended-spectrum β-lactamases (ESBLs) producing E. coli in animals and different methods of identifications from Ado Ekiti, Ekiti State, Nigeria, were investigated. Three hundred and fifty fecal samples, collected from apparently healthy cattle and pigs, were cultured and identified following standard procedures. ESBL phenotypic detection was carried out using combination disc test, double disc synergism test, and ESBL brilliance agar screening. Molecular detection of TEM, SHV,...

  12. Phenotypic and Molecular Characterisation of Extended-Spectrum Beta-Lactamase Producing Escherichia coli Obtained from Animal Fecal Samples in Ado Ekiti, Nigeria

    Directory of Open Access Journals (Sweden)

    Olugbenga Adekunle Olowe

    2015-01-01

    Full Text Available Production of extended-spectrum β-lactamases (ESBLs producing E. coli in animals and different methods of identifications from Ado Ekiti, Ekiti State, Nigeria, were investigated. Three hundred and fifty fecal samples, collected from apparently healthy cattle and pigs, were cultured and identified following standard procedures. ESBL phenotypic detection was carried out using combination disc test, double disc synergism test, and ESBL brilliance agar screening. Molecular detection of TEM, SHV, and CTX-M genes was carried out using standard molecular method. One hundred and fourteen E. coli isolates were recovered from the 350 samples processed, out of which 72 (63.2% isolates were positive for ESBLs with multiple resistance to the antibiotics used. Eighty-one (71% isolates were positive for ESBL by combination disc test, 90 (78.9% were positive for double disc synergism test, and 93 (81.6% were positive for ESBL brilliance agar. TEM and CTX-M genes were detected in 48 (42.1% and 51 (44.7% isolates, respectively. SHV gene was not detected in any of the isolates while TEM and CTX-M were detected in 33 (28.9% isolates. This study showed high resistance of E. coli to antibiotics, particularly to the third generation cephalosporins. Regular monitoring and regulated use of antibiotics in livestock should be encouraged.

  13. Biosynthesis of ketomycin. (II) biomimetic model for beta-lactamase catalysis: host-guest interactions in cyclodextrin-penicillin inclusion complex

    International Nuclear Information System (INIS)

    The antibiotic ketomycin is formed from shikimic acid via chorismic acid and prephenic acid. Phenylalanine and 2',5'-dihydrophenylalanine derived from shikimic acid are not intermediates in the biosynthesis. Degradation of ketomycin derived from [1,6-14C]shikimic acid showed that prephenic acid is converted into ketomycin with stereospecific discrimination between the two enantiotopic edges of the ring, the pro-S-R edge giving rise to the C-2', C-3' side of the cyclohexane ring of ketomycin. The resistance of pathogenic bacteria to the action of β-lactam antibiotics is mainly ascribed to their ability to produce β-lactamase to cleave the β-lactam ring. It is essential to understand the molecular nature of β-lactamase-penicillin recognition for designing and formulating more effective β-lactam antibiotics. A biomimetic study of β-lactamase is therefore initiated. To meet the requirements of hydrophobic and serine protease characteristics of β-lactamase, α-cyclodextrin is chosen as a biomimetic model for β-lactamase. The structural specificity and the chemical dynamics of α-cyclodextrin-phenoxymethyl penicillin inclusion complex in solid state and in solution have been determined by IR and NMR spectroscopy. The spectral results strongly indicate that the phenyl portion of the phenoxymethyl penicillin forms a stable inclusion complex with the hydrophobic cavity of α-cyclodextrin in solution as well as in the solid state. Kinetic studies followed by 1HNMR and HPLC analyses under alkaline condition have shown that the α-cyclodextrin mimics the catalytic function of serine of β-lactamase in the stereospecific hydrolysis of the β-lactam ring of phenoxymethyl penicillin

  14. Frequency and Sensitivity of Extended Spectrum Beta-Lactamase Positive Organisms in a Secondary and Tertiary Level Hospital Network in Dhaka

    OpenAIRE

    Shah Md Zahurul Haque Asna; Shameem Akhter; M Mushfequr Rahman; Najib Mohammad; MA Hafez

    2015-01-01

    Background: Extended spectrum β-lactamase (ESBL) positive organisms are now a global health concern including in Bangladesh. These are associated with treatment failure, increased morbidity and mortality and increased health care costs. In this study, frequency of ESBL positive organisms in some health care centres in Dhaka city has been observed and their current status of antibiogram has also been observed. Objective: To observe the current status of antibiogram of ESBL positive organis...

  15. Antibiotics as selectors and accelerators of diversity in the mechanisms of resistance: From the resistome to genetic plasticity in the beta-lactamases world

    Directory of Open Access Journals (Sweden)

    Juan- Carlos eGalán

    2013-02-01

    Full Text Available Antibiotics and antibiotic resistance determinants, natural molecules closely related to bacterial physiology and consistent with an ancient origin, are not only present in antibiotic-producing bacteria. Throughput sequencing technologies have revealed an unexpected reservoir of antibiotic resistance in the environment. These data suggest that co-evolution between antibiotic and antibiotic resistance genes has occurred since the beginning of time. This evolutionary race has probably been slow because of highly regulated processes and low antibiotic concentrations. Therefore to understand this global problem, a new variable must be introduced, that the antibiotic resistance is a natural event, inherent to life. However, the industrial production of natural and synthetic antibiotics has dramatically accelerated this race, selecting some of the many resistance genes present in nature and contributing to their diversification. One of the best models available to understand the biological impact of selection and diversification are -lactamases. They constitute the most widespread mechanism of resistance, at least among pathogenic bacteria, with more than 1000 enzymes identified in the literature. In the last years, there has been growing concern about the description, spread and diversification of -lactamases with carbapenemase activity and AmpC-type in plasmids. Phylogenies of these enzymes help the understanding of the evolutionary forces driving their selection. Moreover, understanding the adaptive potential of -lactamases contribute to exploration the evolutionary antagonists trajectories through the design of more efficient synthetic molecules. In this review, we attempt to analyse the antibiotic resistance problem from new perspectives. From intrinsic and environmental resistomes to the adaptive potential of resistance genes and the driving forces involved in their diversification, in order to provide a global perspective of the resistance problem.

  16. Extended spectrum beta-lactamase (ESBL) genotypes in Escherichia coli from patients at the Landspítali University Hospital in Iceland from 2006-2012

    OpenAIRE

    Hildur Byström Guðjónsdóttir 1983

    2014-01-01

    Introduction. The prevalence of antibiotic resistance is a growing problem and is a major threat to public health. The first ‘class A’ extended-spectrum β-lactamases (ESBLA), that confer resistance to third-generation cephalosporins, were found in the 1980s as point mutations in the parent enzyme. CTX-M is the newest plasmid-mediated ESBLA, first recognized in 1989 and mostly found in Escherichia coli (E. coli) and Salmonella enterica. The most common ESBL in the world is the CTX-M-15 enzyme,...

  17. Novel variant (bla(VIM-4)) of the metallo-beta-lactamase gene bla(VIM-1) in a clinical strain of Pseudomonas aeruginosa.

    Science.gov (United States)

    Pournaras, Spyros; Tsakris, Athanassios; Maniati, Maria; Tzouvelekis, Leonidas S; Maniatis, Antonios N

    2002-12-01

    A Pseudomonas aeruginosa isolate highly resistant to carbapenems was collected from a patient with postsurgical cerebrospinal infection in Greece. The isolate carried a class 1 integron that contained as a sole cassette the gene bla(VIM-4), a novel variant of bla(VIM-1), with one nucleotide difference resulting in a Ser-to-Arg change at amino acid position 175 of the VIM-1 enzyme. This is the first detection of a VIM-1 variant after its appearance in Italy. PMID:12435718

  18. Eradication of methicillin-resistant Staphylococcus aureus and of Enterobacteriaceae expressing extended-spectrum beta-lactamases on a model pig farm.

    Science.gov (United States)

    Schmithausen, Ricarda Maria; Kellner, Sophia Ricarda; Schulze-Geisthoevel, Sophia Veronika; Hack, Sylvia; Engelhart, Steffen; Bodenstein, Isabel; Al-Sabti, Nahed; Reif, Marion; Fimmers, Rolf; Körber-Irrgang, Barbara; Harlizius, Jürgen; Hoerauf, Achim; Exner, Martin; Bierbaum, Gabriele; Petersen, Brigitte; Bekeredjian-Ding, Isabelle

    2015-11-01

    Colonization of livestock with bacteria resistant to antibiotics is considered a risk for the entry of drug-resistant pathogens into the food chain. For this reason, there is a need for novel concepts to address the eradication of drug-resistant commensals on farms. In the present report, we evaluated the decontamination measures taken on a farm contaminated with methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae expressing extended-spectrum β-lactamases (ESBL-E). The decontamination process preceded the conversion from piglet breeding to gilt production. Microbiological surveillance showed that the decontamination measures eliminated the MRSA and ESBL-E strains that were detected on the farm before the complete removal of pigs, cleaning and disinfection of the stable, and construction of an additional stable meeting high-quality standards. After pig production was restarted, ESBL-E remained undetectable over 12 months, but MRSA was recovered from pigs and the environment within the first 2 days. However, spa (Staphylococcus aureus protein A gene) typing revealed acquisition of an MRSA strain (type t034) that had not been detected before decontamination. Interestingly, we observed that a farmworker who had been colonized with the prior MRSA strain (t2011) acquired the new strain (t034) after 2 months. In summary, this report demonstrates that decontamination protocols similar to those used here can lead to successful elimination of contaminating MRSA and ESBL-E in pigs and the stable environment. Nevertheless, decontamination protocols do not prevent the acquisition of new MRSA strains. PMID:26341200

  19. Insight into the effect of inhibitor resistant S130G mutant on physico-chemical properties of SHV type beta-lactamase: a molecular dynamics study.

    Directory of Open Access Journals (Sweden)

    Mohd Hassan Baig

    Full Text Available Bacterial resistance is a serious threat to human health. The production of β-lactamase, which inactivates β-lactams is most common cause of resistance to the β-lactam antibiotics. The Class A enzymes are most frequently encountered among the four β-lactamases in the clinic isolates. Mutations in class A β-lactamases play a crucial role in substrate and inhibitor specificity. SHV and TEM type are known to be most common class A β-lactamases. In the present study, we have analyzed the effect of inhibitor resistant S130G point mutation of SHV type Class-A β-lactamase using molecular dynamics and other in silico approaches. Our study involved the use of different in silico methods to investigate the affect of S130G point mutation on the major physico-chemical properties of SHV type class A β-lactamase. We have used molecular dynamics approach to compare the dynamic behaviour of native and S130G mutant form of SHV β-lactamase by analyzing different properties like root mean square deviation (RMSD, H-bond, Radius of gyration (Rg and RMS fluctuation of mutation. The results clearly suggest notable loss in the stability of S130G mutant that may further lead to decrease in substrate specificity of SHV. Molecular docking further indicates that S130G mutation decreases the binding affinity of all the three inhibitors in clinical practice.

  20. Antimicrobial resistance in invasive non-typhoid Salmonella from the Democratic Republic of the Congo : emergence of decreased fluoroquinolone susceptibility and extended-spectrum beta lactamases

    OpenAIRE

    Octavie Lunguya; Veerle Lejon; Marie-France Phoba; Sophie Bertrand; Raymond Vanhoof; Youri Glupczynski; Jan Verhaegen; Jean-Jacques Muyembe-Tamfum; Jan Jacobs

    2013-01-01

    Background: Co-resistance against the first-line antibiotics ampicillin, chloramphenicol and trimethoprim/sulphamethoxazole or multidrug resistance (MDR) is common in non typhoid Salmonella (NTS). Use of alternative antibiotics, such as fluoroquinolones or third generation cephalosporins is threatened by increasing resistance, but remains poorly documented in Central-Africa. Methodology/Principal findings: As part of a microbiological surveillance study in DR Congo, blood cultures were collec...

  1. Exploring the potential reservoirs of non specific TEM beta lactamase (blaTEM) gene in the Indo-Gangetic region: A risk assessment approach to predict health hazards.

    Science.gov (United States)

    Singh, Gulshan; Vajpayee, Poornima; Rani, Neetika; Amoah, Isaac Dennis; Stenström, Thor Axel; Shanker, Rishi

    2016-08-15

    The emergence of antimicrobial resistant bacteria is an important public health and environmental contamination issue. Antimicrobials of β-lactam group accounts for approximately two thirds, by weight, of all antimicrobials administered to humans due to high clinical efficacy and low toxicity. This study explores β-lactam resistance determinant gene (blaTEM) as emerging contaminant in Indo-Gangetic region using qPCR in molecular beacon format. Quantitative Microbial Risk Assessment (QMRA) approach was adopted to predict risk to human health associated with consumption/exposure of surface water, potable water and street foods contaminated with bacteria having blaTEM gene. It was observed that surface water and sediments of the river Ganga and Gomti showed high numbers of blaTEM gene copies and varied significantly (pantimicrobial resistant bacteria. PMID:27111425

  2. PREVALENCE AND SUSCEPTIBILITY OF EXTENDED SPECTRUM BETA-LACTAMASES IN URINARY ISOLATES OF ESCHERICHIA COLI IN A TERTIARY CARE HOSPITAL, CHENNAI-SOUTH INDIA

    Directory of Open Access Journals (Sweden)

    Dr. Anbumani Narayanaswamy MD PhD

    2011-01-01

    Full Text Available Extended spectrum beta – lactamases (ESBLs are on the rise in hospital settings across the globe. The presence of ESBLs significantly affects the outcome of an infection and poses a challenge to the management of infection worldwide. Therefore, the aim of the present study is to determine the prevalence and susceptibility of extended spectrum beta – lactamase in urinary isolates of Escherichia coli (E.coli in a tertiary care hospital, Chennai-South India. A total of 450 urinary isolates of E.coli were collected over a period of six months from April 2008 to September 2008. Antimicrobial susceptibility testing was determined to commonly used antibiotics using the modified Kirby-Bauer’s disc diffusion method. ESBL detection was done by the screening method of double disc synergy test and then confirmed by the phenotypic confirmatory test with combination disc as recommended by the Clinical Laboratory Standards Institute (CLSI and the minimum inhibitory concentration (MIC method using the E test strips (AB Biodisk,Sweden - as per manufacturer’s instructions. The prevalence of E.coli ESBL was 60%. The ESBL producing isolates were significantly resistant (p < 0.01 to ampicillin, trimethoprim / sulfamethoxazole, norfloxacin and nalidixic acid as compared to non-ESBL producers. Multidrug resistance was significantly (p < 0.01 higher (69% in ESBL positive isolates than non-ESBL isolates (21%. Knowledge of the prevalence of ESBL and resistance pattern of bacterial isolates in a geographical area will help the clinicians to formulate the guidelines for antibiotic therapy to avoid inappropriate use of extended spectrum cephalosporins.

  3. Carriage of extended-spectrum beta-lactamase-plasmids does not reduce fitness but enhances virulence in some strains of pandemic E. coli lineages

    Directory of Open Access Journals (Sweden)

    Katharina eSchaufler

    2016-03-01

    Full Text Available Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-cured variants (PCV, and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed towards similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages.

  4. Identification of DHA-23, a Novel Plasmid-mediated and Inducible AmpC beta-Lactamase from Enterobacteriaceae in Northern Taiwan

    Directory of Open Access Journals (Sweden)

    Wen-Shyang eHsieh

    2015-05-01

    Full Text Available Objectives: AmpC β-lactamases are classified as Amber Class C and Bush Group 1. AmpC β-lactamases can hydrolyze broad and extended-spectrum cephalosporins, and are not inhibited by β-lactamase inhibitors such as clavulanic acid. This study was conducted to identify DHA-23, a novel plasmid-mediated and inducible AmpC β-lactamase obtained from Enterobacteriaceae. Methods: A total of 210 carbapenem-resistant Enterobacteriaceae isolates were collected from a medical center (comprising 2 branches in Northern Taiwan during 2009–2012. AmpC β-lactamase genes were analyzed through a polymerase chain reaction using plasmid DNA templates and gene sequencing. The genetic relationships of the isolates were typed using pulsed-field gel electrophoresis following the digestion of intact genomic DNA by using XbaI. Results: Three enterobacterial isolates (one Escherichia coli and 2 Klebsiella pneumoniae were obtained from 3 hospitalized patients. All 3 isolates were resistant or intermediately susceptible to all β-lactams, and exhibited reduced susceptibility to carbapenems. These 3 isolates expressed a novel AmpC β-lactamase, designated DHA-23, approved by the curators of the Lahey website. DHA-23 differs from DHA-1 and DHA-6 by one amino acid substitution (Ser245Ala, exhibiting 2 amino acid changes compared with DHA-7 and DHA-Morganella morganii; 3 amino acid changes compared with DHA-3; 4 amino acid changes compared with DHA-5; and 8 amino acid changes compared with DHA-2 (> 97% identity. This AmpC β-lactamase is inducible using a system involving ampR. Conclusion: This is the first report to address DHA-23, a novel AmpC β-lactamase. DHA-type β-lactamases are continuous threat in Taiwan.

  5. Mossambicus tilapia (Oreochromis mossambicus) collected from water bodies impacted by urban waste carries extended-spectrum beta-lactamases and integron-bearing gut bacteria

    Indian Academy of Sciences (India)

    NACHIKET P MARATHE; SWAPNIL S GAIKWAD; ANKITA A VAISHAMPAYAN; MANDAR H RASANE; YOGESH S SHOUCHE; WASUDEV N GADE

    2016-09-01

    Oreochromis mossambicus (Peters 1852) (Tilapia) is one of the most consumed fish globally. Tilapia thrives well inenvironments polluted by urban waste, which invariably contain antibiotic-resistant bacteria and antibiotic resistance genes(ARGs). Thus, Tilapia surviving in such polluted environments may serve as a potential source for dissemination of ARGs.To investigate this, we isolated bacterial strains from gut of Tilapia found in polluted rivers and lakes near Pune, India, andstudied the prevalence of resistance genes bymolecularmethods. A total of 91 bacterial strains were obtained, which includefish pathogens and human pathogens such as Aeromonas hydrophila, Klebsiella pneumoniae, E. coli, Serratia marcescens,Enterobacter spp. and Shigella spp. Overall the prevalence of class 1 integrons, class 2 integrons, extended-spectrum betalactamases(ESBLs) bla_{CTX-M}, bla_{SHV} and aac(6')-Ib-cr gene was 38%, 24%, 38%, 31% and 31% respectively. Forty-twopercent of the Enterobacteriaceae strains carried bla_{CTX-M} gene, which is a common ESBL gene in clinics. The studydemonstrates that tilapia found in the polluted waters can serve as reservoirs and an alternative route for human exposure toclinically important ARG-carrying bacteria. The consumption and handling of these fish may pose a potential health risk.

  6. DETECTION AND ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF PSEUDOMONAS AERUGINOSA ISOLATES IN VARIOUS CLINICAL SAMPLES WITH SPECIAL REFERENCE TO METALLO BETA LACTAMASE FROM A TERTIARY CARE HOSPITAL IN JAIPUR, INDIA

    Directory of Open Access Journals (Sweden)

    Rakesh Kumar

    2014-04-01

    Conclusions- Colistin and Polymyxin-B are more effective to treat multidrug resistant P.aeruginosa. The early detection of MBL producing P. aeruginosa may help in appropriate anti-Pseudomonal therapy to stop the development and dissemination of multidrug resistance strains. [Natl J Med Res 2014; 4(2.000: 128-131

  7. Reservoirs of antibiotic-resistant enterobacteriaceae among animals sympatric to humans in Senegal: extended-spectrum beta-lactamases in bacteria in a black rat (Rattus rattus)

    Czech Academy of Sciences Publication Activity Database

    Literák, I.; Dolejská, M.; Čížek, A.; Djigo, CH. A. T.; Konečný, Adam; Koubek, Petr

    2009-01-01

    Roč. 3, č. 11 (2009), s. 751-754. ISSN 1996-0808 R&D Projects: GA AV ČR IAA6093404 Institutional research plan: CEZ:AV0Z60930519 Keywords : antibiotic s * resistance * Escherichia * Enterobacter * rat * Senegal Subject RIV: FN - Epidemiology, Contagious Diseases ; Clinical Immunology Impact factor: 0.407, year: 2009 http://www.academicjournals.org/ajmr/PDF/Pdf2009/Nov/Literak%20et%20al.pdf

  8. Impact of empirical treatment in extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella spp. bacteremia. A multicentric cohort study

    Directory of Open Access Journals (Sweden)

    Peralta Galo

    2012-10-01

    Full Text Available Abstract Background The objective of this study is to analyze the factors that are associated with the adequacy of empirical antibiotic therapy and its impact in mortality in a large cohort of patients with extended-spectrum β-lactamase (ESBL - producing Escherichia coli and Klebsiella spp. bacteremia. Methods Cases of ESBL producing Enterobacteriaceae (ESBL-E bacteremia collected from 2003 through 2008 in 19 hospitals in Spain. Statistical analysis was performed using multivariate logistic regression. Results We analyzed 387 cases ESBL-E bloodstream infections. The main sources of bacteremia were urinary tract (55.3%, biliary tract (12.7%, intra-abdominal (8.8% and unknown origin (9.6%. Among all the 387 episodes, E. coli was isolated from blood cultures in 343 and in 45.71% the ESBL-E was multidrug resistant. Empirical antibiotic treatment was adequate in 48.8% of the cases and the in hospital mortality was 20.9%. In a multivariate analysis adequacy was a risk factor for death [adjusted OR (95% CI: 0.39 (0.31-0.97; P = 0.04], but not in patients without severe sepsis or shock. The class of antibiotic used empirically was not associated with prognosis in adequately treated patients. Conclusion ESBL-E bacteremia has a relatively high mortality that is partly related with a low adequacy of empirical antibiotic treatment. In selected subgroups the relevance of the adequacy of empirical therapy is limited.

  9. Antimicrobial resistance in faecal Escherichia coli isolates from farmed red deer and wild small mammals. Detection of a multiresistant E. coli producing extended-spectrum beta-lactamase.

    Science.gov (United States)

    Alonso, C A; González-Barrio, D; Tenorio, Carmen; Ruiz-Fons, F; Torres, C

    2016-04-01

    Eighty-nine Escherichia coli isolates recovered from faeces of red deer and small mammals, cohabiting the same area, were analyzed to determine the prevalence and mechanisms of antimicrobial resistance and molecular typing. Antimicrobial resistance was detected in 6.7% of isolates, with resistances to tetracycline and quinolones being the most common. An E. coli strain carrying blaCTX-M-1 as well as other antibiotic resistant genes included in an unusual class 1 integron (Intl1-dfrA16-blaPSE-1-aadA2-cmlA1-aadA1-qacH-IS440-sul3-orf1-mef(B)Δ-IS26) was isolated from a deer. The blaCTX-M-1 gene was transferred by conjugation and transconjugants also acquired an IncN plasmid. This strain was typed as ST224, which seems to be well adapted to both clinical and environmental settings. The phylogenetic distribution of the 89 strains varied depending on the animal host. This work reveals low antimicrobial resistance levels among faecal E. coli from wild mammals, which reflects a lower selective pressure affecting these bacteria, compared to livestock. However, it is remarkable the detection of a multi-resistant ESBL-E. coli with an integron carrying clinically relevant antibiotic-resistance genes, which can contribute to the dissemination of resistance determinants among different ecosystems. PMID:27012919

  10. Evaluation of phenotypic tests for detection of Klebsiella pneumoniae carbapenemase and metallo-beta-lactamase in clinical isolates of Escherichia coli and Klebsiella species

    OpenAIRE

    Kalpana Chauhan; Anita Pandey; Asthana, Ashish K.; Molly Madan

    2015-01-01

    Context: Carbapenemase production is an important mechanism responsible for carbapenem resistance. Aims: Phenotypic detection and differentiation of types of carbapenemase in carbapenem resistant Enterobacteriaceae is important for proper infection control and appropriate patient management. Settings and Design: We planned a study to determine the occurrence of Class A Klebsiella pneumoniae carbapenemase (KPC type) and Class B Metallo-β-lactamase (MBL type) carbapenemase in hospital and commu...

  11. Detection of KPC-2 in a Clinical Isolate of Proteus mirabilis and First Reported Description of Carbapenemase Resistance Caused by a KPC Beta-Lactamase in P. mirabilis

    Science.gov (United States)

    An isolate of Proteus mirabilis recovered from bacterial cultures was shown to be resistant to imipenem, meropenem, and ertapenem by disk diffusion susceptibility testing. Amplification of whole cell and/or plasmid DNA recovered from the isolate using primers specific for the blaKPC carbapenemase g...

  12. Outbreak of Serratia marcescens Coproducing ArmA and CTX-M-15 Mediated High Levels of Resistance to Aminoglycoside and Extended-Spectrum Beta-Lactamases, Algeria.

    Science.gov (United States)

    Batah, Rima; Loucif, Lotfi; Olaitan, Abiola Olumuyiwa; Boutefnouchet, Nafissa; Allag, Hamoudi; Rolain, Jean-Marc

    2015-08-01

    Serratia marcescens is one of the most important pathogens responsible for nosocomial infections worldwide. Here, we have investigated the molecular support of antibiotic resistance and genetic relationships in a series of 54 S. marcescens clinical isolates collected from Eastern Algeria between December 2011 and July 2013. The 54 isolates were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Antibiotic susceptibility testing was performed by disc diffusion and E-test methods. Antibiotic resistance genes were detected by polymerase chain reaction (PCR). The genetic transfer of antibiotic resistance was performed by conjugation using azide-resistant Escherichia coli J53 as the recipient strain, and plasmid analysis was done by PCR-based replicon typing. The relatedness of our isolates was determined by phylogenetic analysis based on partial sequences of four protein-encoding genes (gyrB, rpoB, infB, and atpD) and then compared to MALDI-TOF MS clustering. Thirty-five out of 54 isolates yielded an extended-spectrum β-lactamase (ESBL) phenotype and carried bla(CTX-M-15) (n=32), bla(TEM-1) (n=26), bla(TEM-71) (n=1), bla(SHV-1a) (n=1), and bla(PER-2) (n=12). Among these isolates, we identified a cluster of 15 isolates from a urology unit that coharbored ESBL and the 16S rRNA methyltransferase armA. Conjugation was successful for five selected strains, demonstrating the transferability of a conjugative plasmid of incompatibility group incL/M type. Phylogenetic analysis along with MALDI-TOF clustering likely suggested an outbreak of such isolates in the urology unit. In this study, we report for the first time the co-occurrence of armA methyltransferase with ESBL in S. marcescens clinical isolates in Eastern Algeria. PMID:25884511

  13. Characterization of Two New CTX-M-25-Group Extended-Spectrum beta-Lactamase Variants Identified in Escherichia coli Isolates from Israel

    OpenAIRE

    Vervoort, Jascha; Baraniak, Anna; Gazin, Muriel; Sabirova, Julia; Lammens, Christine; Kazma, Meital; Grabowska, Anna; Izdebski, Radosław; Carmeli, Yehuda; Kumar-Singh, Samir; Gniadkowski, Marek; Goossens, Herman; Malhotra-Kumar, Surbhi

    2012-01-01

    Abstract: Objectives We characterized two new CTX-M-type extended-spectrum β-lactamase (ESBL) variants in Escherichia coli isolates from stool samples of two elderly patients admitted at the Tel Aviv Sourasky Medical Center, Israel. Both patients underwent treatment with cephalosporins prior to isolation of the E. coli strains. Methods ESBLs were detected by the double-disk synergy test and PCR-sequencing of β-lactamase genes. The blaCTX-M genes were cloned into the pCR-BluntII-TOPO vector in...

  14. Molecular characterization and occurrence of extended-spectrum beta-lactamase resistance genes among Salmonella enterica serovar Corvallis from Thailand, Bulgaria and Denmark

    DEFF Research Database (Denmark)

    Archambault, Marie; Petrov, P.; Hendriksen, Rene S.; Asseva, G.; Bangtrakulnonth, A.; Hasman, Henrik; Aarestrup, Frank Møller

    2006-01-01

    confirmed the PFGE-type and was able to further subdivide the strains. Seventeen of these 20 isolates contained also bla(TEM), of which nine representatives were sequenced to bla(TEM-1B), or bla(TEM-1H). One isolate contained bla(CTX-M-15), bla(SHV-2), and bla(TEM-1H), with the bla(CTX-M-15), and bla(TEM-1H...... antimicrobial agents tested, whereas 41 were resistant to three or more antimicrobials. Most resistance was observed among the isolates from Bulgaria. Of the 25 isolates from Bulgaria, 20 displayed resistance to ampicillin and the cephalosporins ceftiofur and cephalothin. All 20 isolates tested negative for bla......(CMY-1), bla(CMY-2), and bla(ACC), but positive for bla(SHV), of which five were sequenced to bla(SHV-2). Plasmid profiling and hybridization revealed that the bla(SHV) gene was located on plasmids of approximately 70 kb. Five plasmid profiles were found among these 20 isolates. The plasmid profiling...

  15. Community-acquired urinary tract infections by extended-spectrum beta-lactamase-producing Enterobacteriaceae in Zenica-Doboj Canton, Bosnia and Herzegovina

    Directory of Open Access Journals (Sweden)

    D. Saric

    2006-08-01

    Full Text Available The aim of this study was to determine the incidence and antimicrobial resistance of ESBL-producing strains in the community-acquired urinary tract infections (CAUTIs, which is necessary for antimicrobial therapy selection. From January 2003 to September 2004, 4,112 consecutive, non-duplicate coliform isolates from CAUTIs were analyzed. Antimicrobial susceptibility testing to fifteen antimicrobials was performed by disc-diffusion method. Double-disk synergy test (DDST with amoxicillin-clavulanat, cefotaxime, ceftazidime, ceftriaxone and aztreonam, and Etest strips with PM/PML (AB Biodisk was performed according to CLSI recommendation in order to detect the ESBL producers. The overall incidence of ESBL producing strains was 2.6% (108/4112, it was significantly higher in males, 8.4% (79/936 than in females, 0.9% (29/3176. The highest prevalence of ESBL producers was noted in the oldest and youngest age group: 4.8% (52/106 and 2.6% (27/1045, respectively. An increase from 2.2% (52/2402 to 3.3% (56/1710, and a shift of ESBL producers toward the age group 0-6 years (1.6% and 3.8%, respectively in this period was observed. The incidence of ESBL producing strains among isolated Klebsiella spp. were 7.8% (83/1060, E. coli 0.7% (18/2561, Citrobacter spp. 0.6% (1/156, Enterobacter spp. 7.7% (3/39 and Proteus spp. 1.0% ( 3/297. Among ESBL producing isolates Klebsiella spp. predominated, 76.9% (83/108, followed by E. coli 16.7% (18/108. ESBL producing strains showed significantly higher resistance rates to all tested antibiotics as compared to to non-ESBL-producers. The increase and shift toward the youngest age group of the ESBL producer incidences is of our concern. Further studies are required to detect ESBL types in terms of highly different geographical dissemination of these isolates.

  16. Sequence analysis of genes mediating extended-spectrum beta-lactamase (ESBL) production in isolates of Enterobacteriaceae in a Lagos Teaching Hospital, Nigeria

    OpenAIRE

    Raji, Muhabat Adeola; Jamal, Wafaa; Ojemeh, Omoh; Rotimi, Vincent Olubunmi

    2015-01-01

    Background Extended-spectrum β-lactamases (ESBLs) in Gram-negative organisms is now a major concern in Enterobacteriaceae worldwide. This study determined a point-prevalence and genetic profiles of ESBL-producing isolates among members of the family Enterobacteriaceae in Lagos State University Teaching Hospital Ikeja, Nigeria. Methods Consecutive non-repetitive invasive multidrug-resistant isolates of the family Enterobacteriaceae obtained over a period of 1 month (October 2011) were studied....

  17. Detection and Molecular Characterization of Escherichia coli Strains Producers of Extended-Spectrum and CMY-2 Type Beta-Lactamases, Isolated from Turtles in Mexico.

    Science.gov (United States)

    Cortés-Cortés, Gerardo; Lozano-Zarain, Patricia; Torres, Carmen; Castañeda, Miguel; Sánchez, Gabriela Moreno; Alonso, Carla A; López-Pliego, Liliana; Mayen, María G Gutiérrez; Martínez-Laguna, Ygnacio; Rocha-Gracia, Rosa Del Carmen

    2016-09-01

    Multidrug-resistant bacteria are a growing problem in different environments and hosts, but scarce information exists about their prevalence in reptiles. The aim of this study was to analyze the resistance mechanisms, molecular typing, and plasmid content of cefotaxime-resistant (CTX(R)) Escherichia coli isolates recovered from cloacal samples of 71 turtles sheltered in a herpetarium in Mexico. CTX(R)-E. coli were recovered in 11 of 71 samples (15.5%), and one isolate/sample was characterized. Extended-spectrum β-lactamase (ESBL)-producing E. coli isolates were detected in four samples (5.6%): two strains carried the blaCTX-M-2 gene (phylogroup D and ST2732) and two contained the blaCTX-M-15 gene (phylogroup B1 and lineages ST58 and ST156). The blaCMY-2 gene was detected by PCR in E. coli isolates of eight samples (9.8%) (one of them also carried blaCTX-M-2); these isolates were distributed into phylogroups A (n = 1), B1 (n = 6), and D (n = 1) and typed as ST155, ST156, ST2329, and ST2732. Plasmid-mediated quinolone resistance (PMQR) genes were detected in five isolates [aac(6')Ib-cr, qnrA, qnrB19, and oqxB]. From three to five replicon plasmids were detected among the strains, being IncFIB, IncI1, IncFrep, and IncK the most prevalent. ESBL or pAmpC genes were transferred by conjugation in four strains, and the blaCTX-M-15 and blaCMY-2 genes were localized in IncFIB or IncI1 plasmids by Southern blot hybridization assays. Class 1 and/or class 2 integrons were detected in eight strains with six different structures of gene cassette arrays. Nine pulsed-field gel electrophoresis patterns were found among the 11 studied strains. To our knowledge, this is the first detection of ESBL, CMY-2, PMQR, and mobile determinants of antimicrobial resistance in E. coli of turtle origin, highlighting the potential dissemination of multidrug-resistant bacteria from these animals to other environments and hosts, including humans. PMID:27482752

  18. First Description of the Extended Spectrum-Beta-Lactamase Gene blaCTX-M-109 in Salmonella Grumpensis Strains Isolated from Neonatal Nosocomial Infections in Dakar, Senegal.

    Directory of Open Access Journals (Sweden)

    Amadou Diop

    Full Text Available Nosocomial infections are very common in African hospitals, particularly in neonatal units. These infections are most often caused by bacteria such as Escherichia coli, Klebsiella spp and Staphylococcus spp. Salmonella strains are rarely involved in nosocomial infections. Here, we report the first description of S. Grumpensis in neonatal infections in Senegal. Seventeen Salmonella strains were isolated from hospitalized infants' stool samples. The following resistance phenotype was described in strains: AMXRTICRCFR FOXRCFXRCTXRCAZRIMPSATMRNARNORRCIPRTMRGMRTERSXTR. All isolates were susceptible to imipenem, 15 out of 17 produced an extended spectrum ß-lactamase (ESBL. blaOXA-1, blaSHV-1, blaTEM-1, blaCTX-M1 genes were detected in strains 8, 13, 5 and 8, respectively. blaCTX-M1 sequencing revealed the presence of blaCTX-M-109. Thirteen of the 17 Salmonella Grumpensis strains were analyzed by PFGE. These 13 isolates belonged to a single pulsotype and were genotypically identical. This is the first report of neonatal S. Grumpensis infections in Senegal, and the first report of blaCTX-M-109 in the genus Salmonella.

  19. Presence of Clostridium difficile and antibiotic and beta-lactamase activities in feces of volunteers treated with oral cefixime, oral cefpodoxime proxetil, or placebo.

    OpenAIRE

    Chachaty, E; Depitre, C; Mario, N; Bourneix, C; Saulnier, P; Corthier, G.; Andremont, A

    1992-01-01

    Three groups of six healthy adult volunteers were randomly assigned to a treatment with 400 mg of oral cefpodoxime proxetil, oral cefixime, or placebo per day for 10 days. Informed consent was obtained from all volunteers. Clostridium difficile was not detected in the feces of any subject before treatment or at any time in the subjects in the placebo group. C. difficile was, however, detected in all subjects treated with cefpodoxime proxetil and in five of six treated with cefixime. Genomic D...

  20. Evaluation of phenotypic tests for detection of Klebsiella pneumoniae carbapenemase and metallo-beta-lactamase in clinical isolates of Escherichia coli and Klebsiella species

    Directory of Open Access Journals (Sweden)

    Kalpana Chauhan

    2015-01-01

    Full Text Available Context: Carbapenemase production is an important mechanism responsible for carbapenem resistance. Aims: Phenotypic detection and differentiation of types of carbapenemase in carbapenem resistant Enterobacteriaceae is important for proper infection control and appropriate patient management. Settings and Design: We planned a study to determine the occurrence of Class A Klebsiella pneumoniae carbapenemase (KPC type and Class B Metallo-β-lactamase (MBL type carbapenemase in hospital and community. Materials and Methods: Clinical isolates of Escherichia coli and Klebsiella species and simultaneously evaluate different phenotypic methods for detection of carbapenemases. Results: It was observed that 20.72% clinical isolates of E. coli and Klebsiella spp. were resistant to carbapenem on screening of which, 14.64% were E. coli and 29.69% were Klebsiella spp. Using phenotypic confirmatory tests the occurrence of carbapenemase production was found to be 87.01% in E. coli and 91.51% in Klebsiella spp. using both modified Hodge test (MHT and combined disk test (CDT using imipenem-ethylenediaminetetraacetic acid. Conclusions: Both MBL and KPC type carbapenemases were seen among clinical isolates of E. coli and Klebsiella spp. CDT is simple, rapid and technically less demanding procedure, which can be used in all clinical laboratories. Supplementing MHT with CDT is reliable phenotypic tests to identify the class A and class B carbapenemase producers.

  1. Carriage of Extended-Spectrum Beta-Lactamase-Plasmids Does Not Reduce Fitness but Enhances Virulence in Some Strains of Pandemic E. coli Lineages

    Science.gov (United States)

    Schaufler, Katharina; Semmler, Torsten; Pickard, Derek J.; de Toro, María; de la Cruz, Fernando; Wieler, Lothar H.; Ewers, Christa; Guenther, Sebastian

    2016-01-01

    Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-“cured” variants (PCV), and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM) assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed toward similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages. PMID:27014251

  2. Seawater is a reservoir of multi-resistant Escherichia coli, including strains hosting plasmid-mediated quinolones resistance and extended-spectrum beta-lactamases genes

    Directory of Open Access Journals (Sweden)

    IsabelHenriques

    2014-08-01

    Full Text Available The aim of this study was to examine antibiotic resistance (AR dissemination in coastal water, considering the contribution of different sources of faecal contamination. Samples were collected in Berlenga, an uninhabited island classified as Natural Reserve and visited by tourists for aquatic recreational activities. To achieve our aim, AR in Escherichia coli isolates from coastal water was compared to AR in isolates from two sources of faecal contamination: human-derived sewage and seagull faeces. Isolation of E. coli was done on Chromocult agar. Based on genetic typing 414 strains were established. Distribution of E. coli phylogenetic groups was similar among isolates of all sources. Resistances to streptomycin, tetracycline, cephalothin and amoxicillin were the most frequent. Higher rates of AR were found among seawater and faeces isolates, except for last-line antibiotics used in human medicine. Multi-resistance rates in isolates from sewage and seagull faeces (29% and 32% were lower than in isolates from seawater (39%. Seawater AR profiles were similar to those from seagull faeces and differed significantly from sewage AR profiles. Nucleotide sequences matching resistance genes blaTEM, sul1, sul2, tet(A and tet(B, were present in isolates of all sources. Genes conferring resistance to 3rd generation cephalosporins were detected in seawater (blaCTX-M-1 and blaSHV-12 and seagull faeces (blaCMY-2. Plasmid-mediated determinants of resistance to quinolones were found: qnrS1 in all sources and qnrB19 in seawater and seagull faeces. Our results show that seawater is a relevant reservoir of AR and that seagulls are an efficient vehicle to spread human-associated bacteria and resistance genes. The E. coli resistome recaptured from Berlenga coastal water was mainly modulated by seagulls-derived faecal pollution. The repertoire of resistance genes covers antibiotics critically important for humans, a potential risk for human health.

  3. Phenotypic detection and occurrence of extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli at a tertiary Hospital in Trinidad & Tobago

    Directory of Open Access Journals (Sweden)

    Patrick E. Akpaka

    2008-12-01

    Full Text Available The incidence and distribution of ESBL producing microorganisms such as E. coli and K. pneumoniae have been demonstrated and varies in different health care facilities and as well as other countries This study was carried out to determine the frequency of occurrence and the antimicrobial susceptibility pattern of ESBL producing E. coli and K. pneumoniae species from clinical isolates at a tertiary hospital in Trinidad & Tobago. Standard microbiological procedures and automated MicroScan System was used to identify, screen for putative ESBL production and determine antimicrobial susceptibility of 1,118 clinical isolates of Enterobacteriaceae species at the microbiology laboratory of the Eric Williams Medical Science Complex, Trinidad & Tobago over a 36 months period. All ESBL producing isolates flagged by the automated system were further confirmed by E-test method. The E-test confirmed a 15.2% ESBL rate among the K. pneumoniae isolates and 9.3% among the E. coli isolates. There was also a 1.8% rate of ESBL production in K. pneumoniae and 0.2% in E. coli isolates from specimens received from community health facilities into the laboratory. Isolates recovered from the intensive care unit of the hospital had 2.1% E. coli and 8.2% K. pneumoniae ESBL producers. Although all ESBL positive isolates were completely susceptible to imipenem and meropenem; and all positive K. pneumoniae isolates were susceptible to amikacin, there was a low susceptibility of ESBL positive E. coli to the aminoglycosides. However, susceptibility of these ESBL producing isolates to the fluoroquinolones varied. There is a high rate of ESBL production among isolates of E. coli and K. pneumoniae at this hospital that is linked to the extensive inappropriate use of third generation cephalosporins in the country. Further molecular studies are needed to characterize the types of these ESBL prevailing in the country.

  4. [In vitro activity of piperacillin-tazobactam against Klebsiella pneumoniae clinical isolates, producers or not of extended spectrum beta-lactamases].

    Science.gov (United States)

    Alarcón, T; de la Obra, P; López-Hernandez, S; de las Cuevas, C; López-Brea, M

    1999-09-01

    The aim of this study was to determine the in vitro activity of piperacillin-tazobactam against 81 clinical isolates of Klebsiella pneumoniae. The clinical specimens were processed according to standard microbiological procedures and 81 K. pneumoniae isolates were identified using MicroScan Panels following the manufacturer's recommendations. A double disk diffusion method was applied to detect extended spectrum betalactamases (ESBL) (43 isolates were positive and 38 were negative). Minimum inhibitory concentrations (MIC) were determined by an agar dilution technique using Mueller-Hinton. The following antibiotics were studied: piperacillin with 4 mg/l of tazobactam, amoxicillin-clavulanic acid in a 2:1 proportion, cefotaxime, ceftriaxone, cefepime, imipenem and meropenem. The MIC(90) were 16/4 mg/l for piperacillin-tazobactam, 16/8 for amoxicillin-clavulanic acid, 16 for ceftriaxone, 16 for cefotaxime, 4 for cefepime, 0.25 for imipenem and 0.032 for meropenem in ESBL-positive strains. In ESBL-negative strains the MIC90 were as follows: 4/4 mg/l for piperacillin-tazobactam, 8/4 for amoxicillin-clavulanic acid, 0.064 for ceftriaxone, 0.125 for cefotaxime, 0.125 for cefepime, 0.125 for imipenem and 0.016 for meropenem. All betalactams showed excellent in vitro activity against ESBL non-producer K. pneumoniae. Moreover, piperacillin-tazobactam and both carbapenems showed good in vitro activity against EBSL-producer K. pneumoniae. PMID:10878513

  5. Investigation of the presence of extended spectrum beta-lactamases (ESBL in multiresistant strains of E. Coli and salmonella species originated from domestic animals

    Directory of Open Access Journals (Sweden)

    Filipović Irina

    2007-01-01

    Full Text Available Bacterial strains which possess genes to produce ESBL most often are multiresistant and also carry genes responsible for the resistance to most other antibiotics, including aminoglycosides, sulfamethoxazole+trimethoprim and fluoroquinolones. Therefore, practically the biggest contemporary clinical problem are infections of humans and animals caused by ESBL-producing strains of E. coli, Kleibsiella, Enterobacter, Proteus, Serratia, Citrobacter, Salmonella and Shigella species. The investigation of the ESBL presence was completed on multiresistant E. coli and Salmonella strains originating from dogs, cats, cattle, sheep, goats, pigs and poultry. The investigated strains were isolated from ear, skin, vaginal, faecal, urine, egg and eggshell swabs, from healthy and diseased individual animals of various ages and breed categories. The sum of 112 E. coli and 45 Salmonella strains was investigated. All strains resistant to 3 or more antibiotics were categorized as multiresistant, which led to a conclusion that 35 E. coli and 6 Salmonella strains out of all investigated were multiresistant to antibiotics. The largest number of multiresistant E. coli strains was discovered in cattle - 12 in total, and the minimal number in goats and sheep, with two strains each. All multiresistant Salmonella strains belonged to the Salmonella Enteritidis species (S. enterica subsp. enterica serovar Enteritidis. The sum of multiresistant Salmonella strains compared to all investigated strains was relatively low (13.3%, but the resistance prevalence for some antibiotics in these strains was extremely high, for ampicillin and amoxicillin with clavulanic acid as high as 100%, and for tetracycline 83.3%. For the control in this investigation were used ESBL positive E. coli strains originated in human urine specimens. No presence of positive ESBL strains was established. However, when the screening investigation was performed, almost all the strains were suspect, thus a confirmatory test had to be performed for all strains.

  6. [In vitro activity of meropenem and seven other beta-lactam antibiotics against K.pneumoniae and enterobacteriaceae producing beta-lactamases with extended spectrum].

    Science.gov (United States)

    Cavallo, J D; Fabre, R; Crenn, Y; Meyran, M

    1994-05-01

    Meropenem is a broad antibacterial spectrum carbapenem with a good activity on betalactam resistant Gram-negative bacilli. 120 non repetitive strains isolated from clinical samples from 1989 to 1992 were selected: 60 K. pneumoniae, 7 E. coli, 2 E. aerogenes and 1 S. marcescens with extended spectrum betalactamases (23 CTX-1, 18 SHV-2, 5 SHV-3, 16 SHV-4, 4 SHV-5, 3 CTX-1 + SHV-4, 1 CAZ-1), 10 K. pneumoniae with broad spectrum TEM-1 enzyme, and 40 K. pneumoniae with only SHV-1 chromosomal betalactamase. Determination of Minimal inhibitory concentrations (MIC) was done by agar dilution method for meropenem and 7 other betalactams (amoxicillin + clavulanic acid 2 mg/l, piperacillin + tazobactam 4 mg/l, cefoxitin, cefotetan, cefotaxime, ceftazidime, imipenem). All antibiotics except amoxicillin + clavulanic acid are active against strains with constitutive penicillinase. For strains with TEM-1 penicillinase, cephamycins, third generation cephalosporins and carbapenems are active. For strains with different extended spectrum betalactamases only cephamycins and carbapenems are efficious. There is no difference according to the period of isolation: 1989-90 or 1991-92. Meropenem has the best in vitro activity (MIC50 = 0.03 mg/l) for all strains independently of the nature of betalactamase. PMID:7824297

  7. Antibacterial activities of multi drug resistant Myroides odoratimimus bacteria isolated from adult flesh flies (Diptera: Sarcophagidae) are independent of metallo beta-lactamase gene Atividades antibacterianas de Myroides odoratimimus isolada de moscas varejeiras adultas (Diptera: Sarcophagidae) são independentes do gene metalo beta lactamase

    OpenAIRE

    M.S. Dharne; A. K. Gupta; Rangrez, A.Y.; H.V. Ghate; Patole, M.S.; Shouche, Y S

    2008-01-01

    Flesh flies (Diptera: Sarcophagidae) are well known cause of myiasis and their gut bacteria have never been studied for antimicrobial activity against bacteria. Antimicrobial studies of Myroides spp. are restricted to nosocomial strains. A Gram-negative bacterium, Myroides sp., was isolated from the gut of adult flesh flies (Sarcophaga sp.) and submitted to evaluation of nutritional parameters using Biolog GN, 16S rRNA gene sequencing, susceptibility to various antimicrobials by disc diffusio...

  8. Sequence of pNL194, a 79.3-kilobase IncN plasmid carrying the blaVIM-1 metallo-beta-lactamase gene in Klebsiella pneumoniae.

    Science.gov (United States)

    Miriagou, V; Papagiannitsis, C C; Kotsakis, S D; Loli, A; Tzelepi, E; Legakis, N J; Tzouvelekis, L S

    2010-10-01

    The nucleotide sequence of pNL194, a VIM-1-encoding plasmid, is described in this study. pNL194 (79,307 bp) comprised an IncN-characteristic segment (38,940 bp) and a mosaic structure (40,367 bp) including bla(VIM-1), aacA7, aadA1, aadA2, dfrA1, dfrA12, aphA1, strA, strB, and sul1. Tn1000 or Tn5501 insertion within fipA probably facilitated recruitment of additional mobile elements carrying resistance genes. PMID:20660690

  9. Quantitative analysis of the IgG and IgG subclass immune responses to chromosomal Pseudomonas aeruginosa beta-lactamase in serum from patients with cystic fibrosis by western blotting and laser scanning densitometry

    DEFF Research Database (Denmark)

    Petersen, T D; Ciofu, O; Pressler, T; Giwercman, B; Pedersen, S S; Høiby, N

    1996-01-01

    IgG1, 79% had IgG4, 56% IgG2, and only 16% of the patients had IgG3 a beta ab. The IgG1 and IgG4 a beta ab appeared first, and more than 50% of the patients were IgG1 and IgG4 a beta ab positive within 2-3 years of the onset of infection, but IgG2 positivity only appeared after seven years and IgG3...... remained absent from most of the patients. The median a beta ab levels increased during chronic infection: 100-fold for IgG1, 22-fold for IgG2, and 45-fold for IgG4. A 16-fold increase in the IgG3 a beta ab levels was detected in the six patients who developed IgG3 a beta ab. In the first four years of the...... chronic infection the a beta ab titres were higher in patients with good lung function than in those with poor lung function. CONCLUSIONS: The association of a weak IgG3 and a strong IgG4 a beta ab response suggests that the contribution of a beta ab antibodies to lung diseases mediated by immune...

  10. 48株流感嗜血杆菌耐药性分析及β-内酰胺酶基因检测%Analysis on antimicrobial resistance and beta-lactamases gene detection of 48 haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    桂和翠; 王中新; 沈继录

    2012-01-01

    目的 了解本地区流感嗜血杆菌的分布及耐药性,为指导临床合理用药提供依据.方法 k-B法进行药敏试验,玻片法测定β-内酰胺酶.PCR扩增TEM及ROB型β-内酰胺酶基因.结果 48株流感嗜血杆菌主要分布于呼吸内科和门诊.对复方新诺明、四环素和氨苄西林耐药率分别为62.50%、35.42% 和22 92%;阿莫西林/克拉维酸钾、氨曲南耐药率为12.50%;氨苄西林/舒巴坦、头孢噻肟、头孢拉定、头孢曲松、阿奇霉素、氯霉素耐药率为8.33%;头孢吡肟、头孢呋辛、环丙沙星、左氧氟沙星耐药率为6.25%.哌拉西林/三唑巴坦、亚胺培南敏感率高为100%.10株氨苄西林耐药菌株均产β-内酰胺酶,产酶率为20.83%,且均检测到TEM基因.结论 复方新诺明和四环素已不再适于临床治疗流感嗜血杆菌引起的感染.氨苄西林仍可作为临床经验用药.哌拉西林/三唑巴坦和亚胺培南抗菌活性高,可望作为治疗耐氨苄西林流感嗜血杆菌感染的理想用药.喹诺酮类药物耐药率高,应引起重视.流感嗜血杆菌对氨苄西林耐药的主要机制为产TEM型β-内酰胺酶.%To investigate antimicrobial resistance and genotypes of β-lactamase of in this erea , and guide clinical rational drug use effectively. Methods Kirby-Bauer method was applied for the drug susceptibility test and nitrocefin slide test was used to detect β-lactamase. The genotypes of β-lactamase were detected by PCR. Results A total of 48 strains of haemophilus influenzae were mainly distributed in department of respiratory and outpatient service. The resistant straint to compound sulf-amethoxazole, ampicillin and tetrocycline were 62. 50% , 35. 42% ,22. 92% respectively, the resistant rate to amoxicillin-clavulanic acid, ceftriaxome, aztreonam, ciprofloxa-cin and levofloxacin was 12.50% ,the resistant rate to ampicillin-sulbactam, cefotaxime, ceftazidime, cefuroxime, azithromycin, chlorampheniol was 8. 33% ;the resistant rate to cefepime, ceftriaxome, ciprofloxacin and levofloxacin was 6. 25%. The hemophilus influenzae were not resistant to piperacillin tazobatam and imipenem. The 10 resistant strains to ampicillin all produced β-lactamases. The prevalence of β-lactamase was 20. 83% . TEM gene was detected in all genotypes of β-lactamases-producing 10 strains. Conclusion The compound sulfamethoxazole and tetrocycline should not be used as empirical treatment of haemophilus influenzae infection. The ampicillin could be used as empirical treatment. The piperacillin-tazobatam and imipenem could be chosen ideally in ampicillin resistant straints. The more attentions should be paid to the resistance of haemophilus influenzae to Quinolone drugs. TEM type of β-lactamase production is the main ampicillin-resistant mechanism of the tested strain.

  11. Caracterización de Klebsiella pneumoniae productora de la beta-lactamasa SHV-5, en una unidad de cuidados intensivos Characterization of SHV-5 beta-lactamase-producing Klebsiella pneumoniae in an intensive care unit

    OpenAIRE

    Verónica Andrade; Jesús Silva

    2004-01-01

    OBJETIVO: Caracterizar molecularmente los aislamientos de Klebsiella pneumoniae obtenidos de pacientes pediátricos y del personal de salud en la unidad de cuidados intensivos de un hospital de tercer nivel de atención en la Ciudad de México, Distrito Federal. MATERIAL Y MÉTODOS: Se analizaron 15 aislamientos de Klebsiella pneumoniae colectadas de un brote durante el mes de junio de 1996, ocho de pacientes y siete de personal del Hospital Infantil de México. Los aislamientos fueron caracteriza...

  12. Characterization of Extended spectrum beta-lactamases (ESBL)-producing Escherichia coli obtained from Danish pigs, pig farmers and their families from farms with high or no consumption of 3rd or 4th generation cephalosporins

    DEFF Research Database (Denmark)

    Hammerum, Anette M.; Larsen, Jesper; Dalhoff Andersen, Vibe; Lester, Camilla H; Skytte, Timmy S. Skovgaard; Hansen, Frank; Olsen, Stefan S.; Mordhorst, Hanne; Skov, Robert L.; Aarestrup, Frank Møller; Agersø, Yvonne

    2014-01-01

    ). Furthermore, transferability of blaCTX-M-1 from both human and pig isolates was studied and plasmid incompatibility groups were defined. The volunteers answered a questionnaire including epidemiological risk factors for carriage of ESBL-producing E. coli. Results: ESBL-producing E. coli was detected in pigs...... on 79% of the farms with high consumption of cephalosporins compared with 20% of the pigs on farms with no consumption. ESBL-producing E. coli was detected in 19 of the 195 human participants and all but one had contact with pigs. The genes found in both humans and pigs at the same farms were blaCTX-M......-1 (eight farms), blaCTX-M-14 (one farm) and blaSHV-12 (one farm). At four farms ESBL-producing E. coli isolates with the same CTX-M enzyme, phylotype, PFGE type and MLST type were detected in both pigs and farmers. The majority of the plasmids with blaCTX-M-1 were transferable by conjugation and...

  13. Usage of antimicrobials on 60 dairy farms in Northern Germany and characterization of Methicillin-resistant staphylococcus aureus (MRSA) and extended spectrum beta-lactamases producing Escherichia coli (ESBLs-producing E. coli) isolated from bulk tank milk samples

    OpenAIRE

    Kreausukon, Khwanchai

    2011-01-01

    Das Ziel der Studie war, Informationen über den Einsatz von Antibiotika in deutschen Milchkuhherden zu sammeln. Zudem sollte auf das Vorkommen von MRSA und ESBL-produzierenden E. coli in Tankmilchproben untersucht werden. Fragebögen wurden unter den Herdenmanagern von 60 norddeutschen Betrieben (Herdengröße von 25 bis 3000 Tiere) verteilt, die auf freiwilliger Basis an den Untersuchungen teilnahmen. Tankmilchproben wurden in den Betrieben einmalig entnommen und auf das Vorkommen von MRSA u...

  14. Fenótipos de betalactamases em Klebsiella pneumoniae de hospital de emergência de Porto Alegre Phenotypes of beta-lactamases in Klebsiella pneumoniae from emergency hospital of Porto Alegre

    OpenAIRE

    Gabriela Meyer; Simone Ulrich Picoli

    2011-01-01

    INTRODUÇÃO: Mundialmente, o crescente aumento de infecções hospitalares gera a necessidade de estudos na área de resistência bacteriana. Uma das bactérias Gram-negativas de maior prevalência no ambiente nosocomial é a Klebsiella pneumoniae, capaz de expressar uma diversidade de enzimas de resistência, justificando seu monitoramento constante. OBJETIVO: Analisar a frequência fenotípica das enzimas betalactamases de espectro estendido (ESBL), cromossômicas (AmpC) plasmidial, metalobetalactamase...

  15. Epidemiologia das betalactamases de espectro estendido no Brasil: impacto clínico e implicações para o agronegócio Epidemiology of extended-spectrum beta-lactamases in Brazil: clinical impact and implications for agribusiness

    OpenAIRE

    Ketrin Cristina da Silva; Nilton Lincopan

    2012-01-01

    A emergência e a disseminação de bactérias produtoras de betalactamases de espectro estendido (ESBL) têm sido retratadas como grande problema de saúde pública, especialmente no que diz respeito a patógenos associados às infecções relacionadas com a assistência à saúde (IRAS). No Brasil, a maior preocupação inclui as altas taxas de resistência a Klebsiella pneumoniae e Escherichia coli, embora as ESBL estejam amplamente disseminadas entre os membros da família Enterobacteriaceae e sejam descri...

  16. Host-Specific Patterns of Genetic Diversity among IncI1-I gamma and IncK Plasmids Encoding CMY-2 beta-Lactamase in Escherichia coli Isolates from Humans, Poultry Meat, Poultry, and Dogs in Denmark

    DEFF Research Database (Denmark)

    Hansen, Katrine Hartung; Bortolaia, Valeria; Nielsen, Christine Ahl;

    2016-01-01

    MLST), restriction fragment length polymorphism (RFLP), and sequencing of selected bla(CMY-2)-harboring plasmids. MLST revealed high strain diversity, with few E. coli lineages occurring in multiple host species and sample types. bla(CMY-2) was detected on plasmids in 83 (89%) isolates. Most (75%) of the plasmids...

  17. THE CONTINUING CHALLENGE OF ESBLS

    OpenAIRE

    Perez, Federico; Endimiani, Andrea; Hujer, Kristine M.; Robert A Bonomo

    2007-01-01

    Since their first description more than twenty years ago, Escherichia coli and Klebsiella pneumoniae possessing extended-spectrum class A beta-lactamases (ESBLs) continue to thwart our best clinical efforts. In the “early years” the most common beta-lactamases were of the TEM and SHV varieties. Now, CTX-M enzymes are being discovered though out the world and are becoming the most prevalent beta-lactamases found in clinical isolates. The Klebsiella pneumoniae carbapenemases (KPC) (ESBL type en...

  18. Synergistic effects of dicloxacillin or clavulanic acid in combination with penicillin G or cephalothin against Yersinia enterocolitica.

    OpenAIRE

    Jimenez-Valera, M; Ruiz-Bravo, A; Ramos-Cormenzana, A.

    1986-01-01

    Cultures of Yersinia enterocolitica grown at 22 degrees C produced beta-lactamases, whereas cultures grown at 37 degrees C produced these enzymes much less effectively. Both dicloxacillin and clavulanic acid inhibited the beta-lactamase activity of bacterial crude extracts and potentiated the activity of penicillin G or cephalothin against 14 Y. enterocolitica strains. It appeared that the beta-lactamase activity present in Y. enterocolitica cells grown at 37 degrees C was great enough to pla...

  19. RELATIVE SPECIFICITIES OF A SERIES OF BETA-LACTAM-RECOGNIZING ENZYMES TOWARDS THE SIDE-CHAINS OF PENICILLINS AND OF ACYCLIC THIOLDEPSIPEPTIDES

    NARCIS (Netherlands)

    SOTO, G; ADACHI, H; VANDERLINDEN, MPG; KECK, W; PRATT, RF

    1994-01-01

    In an attempt to understand more: of the subtle differences between bacterial beta-lactamases and DD-peptidases. comparisons have been made between the specificities of these enzymes towards the phenylacetyl side chain, generally thought to be favoured by beta-lactamases, and the NN'-diacetyl-L-lysy

  20. Binding of benzylpenicillin to metallo-ß-lactamase

    DEFF Research Database (Denmark)

    Olsen, Lars; Rasmussen, T; Hemmingsen, L;

    2004-01-01

    Metallo-beta-lactamases are bacterial enzymes that may function with either one or two zinc ions bound in the active site. In this work, the binding of benzylpenicillin to mono-zinc metallo-beta-lactamase from Bacillus cereus has been investigated in a docking procedure applying a combined quantu...

  1. Lactam hydrolysis catalyzed by mononuclear metallo-ß-bactamases

    DEFF Research Database (Denmark)

    Olsen, Lars; Antony, J; Ryde, U;

    2003-01-01

    Two central steps in the hydrolysis of lactam antibiotics catalyzed by mononuclear metallo-beta-lactamases, formation of the tetrahedral intermediate and its breakdown by proton transfer, are studied for model systems using the density functional B3LYP method. Metallo-beta-lactamases have two met...

  2. Surveillance on Drug Resistance Genes Associated with Extended-spectrum Beta-lactamases in Escherichia coli and Klebsiella neumoniae%产超广谱β-内酰胺酶的大肠埃希菌和肺炎克雷伯菌相关耐药基因监测

    Institute of Scientific and Technical Information of China (English)

    何晓锋; 刘芳; 曹晋桂; 张虎; 吴镝; 焦力群; 马文杰

    2010-01-01

    目的 了解产超广谱β-内酰胺酶(ESBLs)菌中常见的β-内酰胺酶基因的分布情况.方法 采用聚合酶链反应(PCR)检测了77株产ESBLs的大肠埃希菌和肺炎克雷伯菌中TEM、SHV和CTX-M等三种β-内酰胺酶基因,并进行了DNA测序.结果 50株产ESBLs的大肠埃希菌中,41株TEM基因阳性(82.0%),7株SHV基因阳性(14.0%),16株CTX-M基因阳性(32.0%),21株菌两种以上基因型同时阳性.27株产ESBLs的肺炎克雷伯菌中,21株TEM基因阳性(77.8%),24株SHV基因阳性(88.9%),3株CTX-M基因阳性(11.1%),16株菌两种以上基因型同时阳性.结论 产ESBLs的大肠埃希菌和肺炎克雷伯菌中携带多种β-内酰胺酶基因,且以TEM和SHV型较为流行.

  3. Extended-spectrum beta-lactamase-producing Shiga toxin gene (stx1)-positive Escherichia coli O91:H14 carrying blaCTX-M-15 on an IncI1-ST31 plasmid isolated from a human patient in Germany.

    Science.gov (United States)

    Arvand, Mardjan; Bettge-Weller, Gudrun; Fruth, Angelika; Uphoff, Helmut; Pfeifer, Yvonne

    2015-05-01

    In 2011, the Shiga toxin- and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli O104:H4 caused a serious outbreak of gastroenteritis in Germany. This strain carried bla(CTX-M-15) and bla(TEM-1) on an IncI1-ST31 plasmid. During screening of individuals at risk for acquisition of the epidemic E. coli O104:H4, we isolated another ESBL-producing and Shiga toxin-positive E. coli belonging to serotype O91:H14 from feces of a human patient. Interestingly, the patient also carried a further ESBL-producing but Shiga toxin-negative E. coli. Both strains harbored bla(CTX-M-15) and bla(TEM-1) on an IncI1-ST31 plasmid, which was indistinguishable regarding size and plasmid restriction pattern from the plasmid of the epidemic E. coli O104:H4 strain. The patient had traveled to India 6 months prior to the isolation of the E. coli strains. This is the first report of an ESBL-producing, Shiga toxin-positive E. coli of serogroup O91. Our data suggest a high propensity of the IncI1-ST31 plasmid to spread in the human and/or animal population. PMID:25801683

  4. Multiple chromosomes of Azotobacter vinelandii.

    OpenAIRE

    1989-01-01

    The number of copies of the genes leuB, nifH, nifD, and nifK per cell of Azotobacter vinelandii has been determined to be about 80. A beta-lactamase gene was integrated into the A. vinelandii chromosome by single-point crossover. Subsequently, we have been able to detect nearly 80 copies of this beta-lactamase gene per cell of A. vinelandii when cultured for a large number of generations in the presence of ampicillin. The multiple copies of the beta-lactamase gene do not seem to be present on...

  5. Killing curve activity of ciprofloxacin is comparable to synergistic effect of beta-lactam-tobramycin combinations against Haemophilus species endocarditis strains

    DEFF Research Database (Denmark)

    Westh, H; Frimodt-Møller, N; Gutschik, E;

    1992-01-01

    Nine Haemophilus species strains, all beta-lactamase negative, isolated from patients with endocarditis were tested in killing curve experiments. Antibiotics used were penicillin, amoxicillin, aztreonam alone and in combination with tobramycin, as well as ciprofloxacin alone. Synergism between beta...

  6. Disease: H00301 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available ons caused by extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae: risk factors, molecular epidemiology, and clinical...bsiella pneumoniae hospital acquired bacteremia. Risk factors and clinical outcome. Saudi Med J 25:1871-6 (2004) ...

  7. Evaluation of restoration of sensitivities of resistant staphylococcus aureus isolates by using cefuroxime and clavulanic acid in combination

    International Nuclear Information System (INIS)

    The present study was planned to observe the activity of cefuroxime, a second generation cephalosporin after combining it with a beta-lactamase inhibitor calvulanic acid. The study was conducted to evaluate the restoration or increase in sensitivity of beta-lactamase producing isolates of Staphylococcus aureus. Staphylococcus aureus were identified by standard procedures. For beta-lactamase detection chromogenic Nitrocefin impregnated sticks were used. The sensitivity of the bacteria to the antibiotic disks was measured by disk diffusion method using standard zone diameter criteria given by National Committee of Clinical Laboratory Standards. The disks of cefuroxime with clavulanic acid had developed larger zones of inhibition. The activity of cefuroxime against Staphylococcus areus was significantly increased by clavulanic acid. Clavulanic acid if used in combination with cefuroxime, can improve the antimicrobial activity of cefuroxime against beta - lactamase producing Staphylococcus aureus. (author)

  8. Some Virulence Factors of Staphylococci Isolated From Wound and Skin Infections in Shahrekord, IR Iran

    OpenAIRE

    Azizollah Ebrahimi; Maryam Ghasemi; Bahram Ghasemi

    2014-01-01

    Background:: Staphylococci release a large number of enzymes. Some of these, such as coagulase, beta- lactamase, hemolysins and biofilms are considered indices of pathogenicity. Objectives:: The aim of the current study was based on the isolation and identification of Staphylococcus aureus and coagulase negative Staphylococci (CNS) strains from various skin lesions and examining their biofilms, beta- lactamase, hemolysins production and antibiotic resistance pattern. Materials and Met...

  9. Diversity of β-lactamase-encoding genes in Escherichia coli strains isolated from food-producing, companion and zoo animals in Portugal

    OpenAIRE

    Clemente, Lurdes; Correia, Ivone; Albuquerque, Teresa; Geraldes, Margarida; Matos, Filipa; Themudo, Patrícia; Manageiro, Vera; Jones-Dias, Daniela; Ferreira, Eugénia; Caniça, Manuela

    2012-01-01

    A rapid development of plasmid-mediated resistance to extended-spectrum cephalosporins has been observed in Enterobacteriaceae worldwide, predominantly due to the dissemination of extended-spectrum beta-lactamases (ESBL) and plasmid-mediated AmpC beta-lactamases (PMAB). The aim of the present study was to evaluate the extension of ESBL- and PMAB-producing E. coli strains isolated from different animal origins in Portugal. For surveillance purposes, 376 E. coli isolates identified at Natio...

  10. Susceptibilities of 540 anaerobic gram-negative bacilli to amoxicillin, amoxicillin-BRL 42715, amoxicillin-clavulanate, temafloxacin, and clindamycin.

    OpenAIRE

    Appelbaum, P. C.; Spangler, S K; Shiman, R; Jacobs, M R

    1992-01-01

    Agar dilution MIC testing of amoxicillin, amoxicillin-BRL 42715, amoxicillin-clavulanate, temafloxacin, and clindamycin against 496 beta-lactamase-producing anaerobic gram-negative rods revealed MICs for 90% of the strains tested of 256.0 (amoxicillin), 2.0 (amoxicillin-BRL 42715 and amoxicillin-clavulanate), and 4.0 (temafloxacin and clindamycin) microgram/ml. Amoxicillin, temafloxacin, and clindamycin inhibited all 44 beta-lactamase-negative strains (MICs for 90% of the strains tested, less...

  11. Imported PER-1 producing Pseudomonas aeruginosa, PER-1 producing Acinetobacter baumanii and VIM-2-producing Pseudomonas aeruginosa strains in Hungary

    OpenAIRE

    Nagy Károly; Katona Katalin; Juhász Zsuzsa; Szentandrássy Julia; Szabó Dora; Rókusz László

    2008-01-01

    Abstract Introduction Pseudomonas aeruginosa and Acinetobacter baumanii are important nosocomial pathogens with wide intrinsic resistance. However, due to the dissemination of the acquired resistance mechanisms, such as extended-spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) production, multidrug resistant strains have been isolated more often. Case presentation We report a case of a Hungarian tourist, who was initially hospitalized in Egypt and later transferred to Hungary. ...

  12. Cloning and expression in Escherichia coli of the Azospirillum brasilense Sp7 gene encoding ampicillin resistance.

    OpenAIRE

    Verreth, C; Cammue, B; Swinnen, P; Crombez, D; Michielsen, A; Michiels, K.; Gool, A. van; Vanderleyden, J.

    1989-01-01

    The Azospirillum brasilense ATCC 29145 gene coding for beta-lactamase was cloned in Escherichia coli. The gene was expressed in E. coli from its own promoter as a 30-kilodalton protein, conferring resistance to high levels of beta-lactam antibiotics. The DNA sequence containing the beta-lactamase gene was found to be highly amplified in the Azospirillum genome, scattered in the chromosomal as well as in the plasmidic DNA.

  13. Incidence of temonera, sulphuhydryl variables and cefotaximase genes associated with β-lactamase producing escherichia coli in clinical isolates

    Directory of Open Access Journals (Sweden)

    Ibeh Nnana Isaiah

    2011-01-01

    Full Text Available Background: The occurrence of the different types of Extended spectrum beta Lactamase producing Escherichia coli with the, Sulphurhydryl variable, Temonera and the Cefotaximase have been on the rise Aim: The study was to determine the prevalence of extended spectrum beta lactamase gene resistance across the clinical isolates of hospitalized patients. Materials and Method: Three hundred and fifty isolates of Escherichia coli were received from different clinical specimens. The susceptibility profile of the isolates against 10 different antibiotics was examined, the MICs (Minimum Inhibitory Concentration for ceftazidime were also determined using micro-broth dilution assay. Isolates showing MIC ≥ 6 μg/ml for ceftazidime were screened for ESBL (PCTphenotypic confirmatory test and subjected to PCR (polymerase chain reaction to further. Results: By disk diffusion test, there was resistance to ceftazidime and cefotaxime were 180(51.4% and 120 (34.2% respectively. However, all strains were susceptible to imipenem. 250 isolates showed MICs≥ 6 μg/ml for ceftazidime of which 180 (72% were positive for extended spectrum beta lactamase. The prevalence of Sulphurhydryl variable, Temonera and the Cefotaximase among these isolates were 17.1%, 6.6% and 17%, respectively. Conclusion: For the identification of extended spectrum beta lactamase producing isolates it is recommended that clinical laboratories adopt simple test based on Cinical laboratory standard institute recommendation for confirming extended spectrum beta lactamase production in enterobacteriacea species.

  14. An international multicenter study of antimicrobial consumption and resistance in Staphylococcus aureus isolates from 15 hospitals in 14 countries

    DEFF Research Database (Denmark)

    Westh, Henrik Torkil; Zinn, Christina Scheel; Rosdahl, Vibeke Thamdrup

    2004-01-01

    usage of therapeutical subgroups of antimicrobials varied significantly between hospitals. A positive correlation was found between S. aureus resistance to methicillin (MRSA) and consumption of beta-lactam combinations, between resistance to quinolones and consumption of beta-lactam combinations and...... carbapenems and resistance to aminoglycosides and consumption of beta-lactam combinations. The consumption of beta-lactamase-sensitive antibiotics was negatively correlated to resistance to methicillin, quinolones, and aminoglycosides. Usage of the different antimicrobial therapeutical subgroups was also...... correlated. Consumption of beta-lactamase-sensitive antibiotics (penicillin) was positively correlated to consumption of beta-lactamase-resistant penicillins and negatively correlated to consumption of carbapenems, quinolones, and glycopeptides, whereas consumption of cephalosporins was positively correlated...

  15. Treatment for infections complications of experimental acute radiation sickness with sulacillin, a combined antibiotic

    International Nuclear Information System (INIS)

    The therapeutic efficiency of sulacillin (combination of ampicillin antibiotic with beta-lactamase sulbactam inhibitor) used for prevention and treatment of infections complications of the acute radiation disease (ARD) is considered. It is shown that sulacillin antiinfections effect essentially exceeds the activity of ampicillin by treatment of irradiated mice infected with a beta-lactamase-producing strain of Kl pneumoniae. Inclusion of the sulacillin as a principal antibiotic into the ARD therapeutic scheme provides for the 66.6 % survival of dogs at LD90/45

  16. Pharmacodynamics of Imipenem in Combination with β-Lactamase Inhibitor MK7655 in a Murine Thigh Model

    OpenAIRE

    Mavridou, Eleftheria; Melchers, Ria J. B.; van Mil, Anita C. H. A. M.; Mangin, E.; Motyl, Mary R.; Mouton, Johan W.

    2014-01-01

    MK7655 is a newly developed beta-lactamase inhibitor of class A and class C carbapenemases. Pharmacokinetics (PK) of imipenem-cilastatin (IMP/C) and MK7655 were determined for intraperitoneal doses of 4 mg/kg to 128 mg/kg of body weight. MIC and pharmacodynamics (PD) studies of MK7655 were performed against several beta-lactamase producing Pseudomonas aeruginosa and Klebsiella pneumoniae strains to determine its effect in vitro and in vivo. Neutropenic mice were infected in each thigh 2 h bef...

  17. Sobreexpressão e purificação da carbapenemase SFC-1 produzida por um isolado deserratia fonticola

    OpenAIRE

    Fonseca, Maria Fátima da

    2005-01-01

    A espécie Serratia fonticola inclui organismos que ocorrem naturalmente em águas. A maioria das estirpes expressa uma beta-lactamase cromossomal de espectro alargado de classe A (SFO-1), e uma beta-lactamase do tipo AmpC, específica da espécie. A estirpe S. fonticola UTAD54, isolada de águas de consumo não tratadas no Nordeste de Portugal, mostrou ser resistente a vários antibióticos beta-lactâmicos incluindo os carbapenemos. O genoma da estirpe UTAD54 codifica para uma meta...

  18. Nucleotide sequence and characterization of a carbenicillin-hydrolyzing penicillinase gene from Proteus mirabilis.

    OpenAIRE

    Sakurai, Y.; Tsukamoto, K; Sawai, T

    1991-01-01

    The structural gene of a carbenicillinase was cloned from the chromosomal DNA of Proteus mirabilis GN79. This gene codes for a protein of 270 amino acids. Alignment of the amino acid sequence with those of known beta-lactamases revealed that the enzyme is a novel class A beta-lactamase with a unique conserved triad, RTG. By using a DNA fragment of the structural gene, a lack of cross hybridization was confirmed between the DNA probe and total DNAs from natural isolates of P. mirabilis, sugges...

  19. Design, Synthesis, and Crystal Structures of 6-Alkylidene-2 -Substituted Penicillanic Acid Sulfones as Potent Inhibitors of Acinetobacter baumannii OXA-24 Carbapenemase

    Energy Technology Data Exchange (ETDEWEB)

    Bou, G.; Santillana, E; Sheri, A; Beceiro, A; Sampson, J; Kalp, M; Bethel, C; Distler, A; Drawz, S; et. al.

    2010-01-01

    Class D {beta}-lactamases represent a growing and diverse class of penicillin-inactivating enzymes that are usually resistant to commercial {beta}-lactamase inhibitors. As many such enzymes are found in multi-drug resistant (MDR) Acinetobacter baumannii and Pseudomonas aeruginosa, novel {beta}-lactamase inhibitors are urgently needed. Five unique 6-alkylidene-2{prime}-substituted penicillanic acid sulfones (1-5) were synthesized and tested against OXA-24, a clinically important {beta}-lactamase that inactivates carbapenems and is found in A. baumannii. Based upon the roles Tyr112 and Met223 play in the OXA-24 {beta}-lactamase, we also engineered two variants (Tyr112Ala and Tyr112Ala,Met223Ala) to test the hypothesis that the hydrophobic tunnel formed by these residues influences inhibitor recognition. IC{sub 50} values against OXA-24 and two OXA-24 {beta}-lactamase variants ranged from 10 {+-} 1 (4 vs WT) to 338 {+-} 20 nM (5 vs Tyr112Ala, Met223Ala). Compound 4 possessed the lowest K{sub i} (500 {+-} 80 nM vs WT), and 1 possessed the highest inactivation efficiency (k{sub inact}/K{sub i} = 0.21 {+-} 0.02 {micro}M{sup -1}s{sup -1}). Electrospray ionization mass spectrometry revealed a single covalent adduct, suggesting the formation of an acyl-enzyme intermediate. X-ray structures of OXA-24 complexed to four inhibitors (2.0-2.6 {angstrom}) reveal the formation of stable bicyclic aromatic intermediates with their carbonyl oxygen in the oxyanion hole. These data provide the first structural evidence that 6-alkylidene-2{prime}-substituted penicillin sulfones are effective mechanism-based inactivators of class D {beta}-lactamases. Their unique chemistry makes them developmental candidates. Mechanisms for class D hydrolysis and inhibition are discussed, and a pathway for the evolution of the BlaR1 sensor of Staphylococcus aureus to the class D {beta}-lactamases is proposed.

  20. Presence of ESBL/AmpC-producing Escherichia coli in the broiler production pyramid: a descriptive study.

    OpenAIRE

    Dierikx, C.M.; van der Goot, J. A.; Smith, H E; Kant, A.; Mevius, D.J.

    2013-01-01

    Broilers and broiler meat products are highly contaminated with extended spectrum beta-lactamase (ESBL) or plasmid-mediated AmpC beta-lactamase producing Escherichia coli and are considered to be a source for human infections. Both horizontal and vertical transmission might play a role in the presence of these strains in broilers. As not much is known about the presence of these strains in the whole production pyramid, the epidemiology of ESBL/AmpC-producing E. coli in the Dutch broiler produ...