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Sample records for beta-lactamases

  1. Beta-lactamases in Enterobacteriaceae in broilers

    NARCIS (Netherlands)

    Dierikx, C.M.

    2013-01-01

    Resistance to cephalosprins due to the production of extended spectrum beta-lactamases (ESBLs) or plasmid mediated AmpC beta-lactamases is increasingly found in infections in humans outside the hospital. The genes encoding for these beta-lactamases are located on mobile DNA (plasmids), which can be

  2. Antibodies against chromosomal beta-lactamase

    DEFF Research Database (Denmark)

    Giwercman, B; Rasmussen, J W; Ciofu, Oana

    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...... 1 cephalosporinase. We found a wide range of chromosomal beta-lactamase activity in the sputum samples, with no correlation with basal or induced activity of beta-lactamase expression. The presence of anti-beta-lactamase antibodies in endobronchial sputum could be an important factor in the defense...

  3. 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

  4. Encoded Extended Spectrum Beta-Lactamases Produced

    African Journals Online (AJOL)

    26.1 % Klebsiella spp were positive for extended spectrum beta-lactamases ... issue, and TEM, OXA and SHV type ESBL were the most common genotypes. ... mechanism of action. ..... and Multiplex PCR Screening of AmpC Genes from.

  5. 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 c...... could be a major reason for the persistence of this sessile bacterium in chronic infections....

  6. Beta-lactamases of Mycobacterium tuberculosis and Mycobacterium kansasii.

    Science.gov (United States)

    Segura, C; Salvadó, M

    1997-09-01

    Re-emergence of infectious diseases caused by mycobacteria as well as the emergence of multiresistant strains of Mycobacterium has promoted the research on the use of beta-lactames in the treatment of such diseases. Mycobacteria produce beta-lactamases: M. tuberculosis produces a wide-spectrum beta-lactamase whose behaviour mimicks those of Gram-negative bacteria. M. kansasii produces also beta-lactamase which can be inhibited by clavulanic acid. An overview on beta-lactamases from both species is reported.

  7. Antibiotic resistance patterns and beta-lactamase identification in ...

    African Journals Online (AJOL)

    Children acquire bacteria from their mother during birth,[3,4] and ... Our results revealed high resistance rates to co-trimoxazole (54.0%), penicillin .... the inclusion of a beta-lactamase inhibitor, clavulanic acid. .... Folate pathway inhibitor/.

  8. Strategic Design of an Effective beta-Lactamase Inhibitor

    Energy Technology Data Exchange (ETDEWEB)

    Pattanaik, P.; Bethel, C; Hujer, A; Hujer, K; Distler, A; Taracila, M; Anderson, V; Fritsche, T; Jones, R; et. al.

    2009-01-01

    In an effort to devise strategies for overcoming bacterial beta-lactamases, we studied LN-1-255, a 6-alkylidene-2'-substituted penicillin sulfone inhibitor. By possessing a catecholic functionality that resembles a natural bacterial siderophore, LN-1-255 is unique among beta-lactamase inhibitors. LN-1-255 combined with piperacillin was more potent against Escherichia coli DH10B strains bearing bla(SHV) extended-spectrum and inhibitor-resistant beta-lactamases than an equivalent amount of tazobactam and piperacillin. In addition, LN-1-255 significantly enhanced the activity of ceftazidime and cefpirome against extended-spectrum cephalosporin and Sme-1 containing carbapenem-resistant clinical strains. LN-1-255 inhibited SHV-1 and SHV-2 beta-lactamases with nm affinity (K(I) = 110 +/- 10 and 100 +/- 10 nm, respectively). When LN-1-255 inactivated SHV beta-lactamases, a single intermediate was detected by mass spectrometry. The crystal structure of LN-1-255 in complex with SHV-1 was determined at 1.55A resolution. Interestingly, this novel inhibitor forms a bicyclic aromatic intermediate with its carbonyl oxygen pointing out of the oxyanion hole and forming hydrogen bonds with Lys-234 and Ser-130 in the active site. Electron density for the 'tail' of LN-1-255 is less ordered and modeled in two conformations. Both conformations have the LN-1-255 carboxyl group interacting with Arg-244, yet the remaining tails of the two conformations diverge. The observed presence of the bicyclic aromatic intermediate with its carbonyl oxygen positioned outside of the oxyanion hole provides a rationale for the stability of this inhibitory intermediate. The 2'-substituted penicillin sulfone, LN-1-255, is proving to be an important lead compound for novel beta-lactamase inhibitor design.

  9. Phenotypic Characterization of Multidrug-resistant Escherichia Coli with Special Reference to Extended-spectrum-beta-lactamases and Metallo-beta-lactamases in a Tertiary Care Center.

    Science.gov (United States)

    Shrestha, B; Shrestha, S; Mishra, S K; Kattel, H P; Tada, T; Ohara, H; Kirikae, T; Rijal, B P; Sherchand, J B; Pokhrel, B M

    2015-01-01

    The increasing reports on extended-spectrum-beta-lactamase and metallo-beta-lactamase producing Escherichia coli have addressed a potential threat to global health since it is found to be highly resistance to most of the currently available antibiotics including carbapenems. The present study was aimed to determine the antibiogram of extended-spectrum-beta-lactamase and metallo-beta-lactamase producing MDR E. coli isolates from various clinical samples. This was a cross-sectional study conducted over a period of seven months from December 2013 to July 2014 at bacteriology laboratory of Tribhuvan University Teaching Hospital. A total of 250 clinical specimens (urine, pus, sputum, blood, body fluid, bile, tissue and central venous pressure line tip) were processed from inpatients, with multidrug-resistant Escherichia coli infections. Standard microbiological techniques were used for isolation and identification of the isolates. The presence of extended-spectrum-beta-lactamase was detected by phenotypic confirmatory test recommended by Clinical and Laboratory Standards Institute and imipenem (IMP) /EDTA combined disc method was performed to detect metallo-beta-lactamase mediated resistance mechanism. We found high level of beta lactamase mediated resistance mechanism as part of multidrug resistance. Among 250 MDR isolates, 60% isolates were extended-spectrum-beta-lactamase producers and 17.2% isolates were metallo-beta-lactamase producers. Co-existence of extended-spectrum-beta-lactamase and metallo-beta-lactamase identified in 6.8% isolates. Beta-lactamase mediated resistance mechanisms are accounting very high in the multidrug resistant isolates of E. coli. Therefore, early detection of beta lactamase mediated resistant strains and their current antibiotic susceptibility pattern is necessary to avoid treatment failure and prevent the spread of MDR.

  10. Beta-lactamases in Enterobacteriaceae infections in children.

    Science.gov (United States)

    Moxon, Christopher Alan; Paulus, Stéphane

    2016-07-05

    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. Copyright © 2016 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

  11. Design, synthesis and bioactivity evaluation of tribactam beta lactamase inhibitors.

    Science.gov (United States)

    Copar, Anton; Prevec, Tadeja; Anzic, Borut; Mesar, Tomaz; Selic, Lovro; Vilar, Mateja; Solmajer, Tom

    2002-03-25

    Known carbapenem compounds with inhibitory effect towards beta-lactamase enzymes are formed from bicyclical beta lactam structural scaffolds. On the basis of results from theoretical computational methods and molecular modelling we have designed and developed a synthetic route towards novel, biologically active tricyclic derivatives of carbapenems.

  12. OXA beta-lactamase-mediated carbapenem resistance in Acinetobacter baumannii

    Directory of Open Access Journals (Sweden)

    S M Amudhan

    2011-01-01

    Full Text Available Objectives: Acinetobacter baumannii is a significant pathogen in health care settings. In recent years, an increase in carbapenem resistance among A. baumannii due to Ambler class B metallo-beta-lactamases or class D OXA carbapenamases has been reported. In this study we detected the presence of OXA carbapenamases and coproduction of metallo-beta-lactamases (blaVIM and blaIMP by phenotypic and genotypic methods in carbapenem resistant clinical isolates of Acinetobacter baumannii. Materials and Methods: A total of 116 consecutive, non-duplicate carbapenem resistant A. baumannii isolated from various clinical specimens were included in the study. The modified Hodge test and inhibitor potentiated disk diffusion tests were done for the screening of carbapenamase and metallo-beta-lactamase production, respectively. Polymerase chain reaction (PCR was performed for the detection of OXA (blaOXA 23 like, blaOXA 24 like, blaOXA-51 like and blaOXA-58 like genes and metallo-beta-lactamases (blaVIM and blaIMP genes. Gene sequencing was performed for representative isolates. Results: Among 116 A. baumannii, OXA genes were detected in 106 isolates. BlaOXA 51 like (n = 99 and blaOXA -23 like (n = 95 were the most common and they coexisted in 89 isolates. blaOXA-24 like gene was detected in two isolates of which one also carried blaOXA-51 like and blaOXA-58 like genes. The modified Hodge test was positive in 113 isolates. The metallo-beta-lactamase screening test was positive in 92 isolates. blavim was detected in 54 isolates of which 1 also carried the blaIMP gene. Conclusions: blaOXA-23 like and bla OXA 51 like genes are the most common types of OXA carbapenamases while the blaVIM type is the most common type of metallo-beta-lactamase contributing to carbapenem resistance in clinical isolates of A. baumannii. The coproduction of OXA and metallo-beta-lactamases is not an uncommon phenomenon in A. baumannii.

  13. Phenotypic Characterization of Multidrug-resistant Escherichia Coli with Special Reference to Extended-spectrum-beta-lactamases and Metallo-beta-lactamases in a Tertiary Care Center

    Directory of Open Access Journals (Sweden)

    Basudha Shrestha

    2015-06-01

    Conclusions: Beta-lactamase mediated resistance mechanisms are accounting very high in the multidrug resistant isolates of E. coli. Therefore, early detection of beta lactamase mediated resistant strains and their current antibiotic susceptibility pattern is necessary to avoid treatment failure and prevent the spread of MDR. Keywords: e. coli; extended-spectrum-β-lactamase; metallo-β-lactamase; multidrug-resistance.

  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

    OpenAIRE

    Gupta, V.; Garg, R.; Garg, S.; Chander, J.; 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...

  15. 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

  16. Fecal Carriage of Extended-spectrum Beta-lactamase and AmpC ...

    African Journals Online (AJOL)

    2018-02-07

    Feb 7, 2018 ... beta-lactamase-producing Enterobacteriaceae in community and to investigate cefotaxime-M (CTX-M) genes ... trimethoprim–sulfamethoxazole, and carbapenems were 31.2%, 33.3%, and. 0%, respectively. Conclusion: .... with phenotypic AmpC beta-lactamase were resistant to ceftazidime and CTX and ...

  17. Molecular identification of TEM-116 beta-lactamase gene in isolates ...

    African Journals Online (AJOL)

    Purpose: Purpose: To determine TEM-116 beta-lactamase gene prevalence in drug-resistant Pseudomonas aeruginosa isolates from Pakistan. Methods: Sequence analysis of TEM beta-lactamase isolates and their antibiotic susceptibility patterns were carried out. Quantitative bacteriostatic concentrations for commonly ...

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

    DEFF Research Database (Denmark)

    Hemmingsen, Lars Bo Stegeager; Olsen, L.; Antony, J.

    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...

  19. Extended spectrum beta-lactamases in urinary gram-negative bacilli and their susceptibility pattern

    International Nuclear Information System (INIS)

    Mumtaz, S.

    2008-01-01

    Beta-lactamases of gram-negative bacteria are the most important mechanism of resistance against beta lactams. Two types of beta-lactamases can confer resistance against third generation cephalosporins inducible Chromosomal beta -lactamases and extended-spectrum beta-lactamases. The extended-spectrum beta lactamases producing Strains of Enterobacteriaceae have emerged as a major problem in hospitalized as well as community based infections resulting in range of infections from uncomplicated urinary tract infection to life threatening sepsis. The study was conducted at the Microbiology Department of Fauji Foundation Hospital, Rawalpindi over a period of two years (April 2004-March 2006). Multidrug resistance and extended spectrum beta-lactamases production was studied in 111 enteric Gram-negative bacilli isolated from urine of symptomatic patients (1- 70 years) including males and females from indoor and outdoor patients by using double disc diffusion technique. Prevalence of extended-spectrum beta-lactamases production was seen in 71 (61.2%) enteric gram-negative organisms, the most prevalent gram-negative organism was Klebsiella pneumoniae 40 (71.4%) followed by Escherichia coli 27 (62.8%) and Pseudomonas aeruginosa 3 (25%). The extended-spectrum beta-lactamases producers were more prevalent in indoor patients 63 (88.7%) compared to outdoor patients 8 (11.3%), more in females 43 (60.6%) than males, 28 (39.4%). The extended-spectrum beta-lactamases producing gram-negative rods had more antibiotic-resistant profile than non-producers. All enteric gram negative rods should be tested for the production of extended-spectrum beta-lactamases in routine microbiology laboratory. (author)

  20. Extended Spectrum Beta-lactamases: Definition, Classification and Epidemiology.

    Science.gov (United States)

    Ghafourian, Sobhan; Sadeghifard, Nourkhoda; Soheili, Sara; Sekawi, Zamberi

    2015-01-01

    Extended spectrum beta-lactamases (ESBLs) are defined as enzymes produced by certain bacteria that are able to hydrolyze extended spectrum cephalosporin. They are therefore effective against beta-lactam antibiotics such as ceftazidime, ceftriaxone, cefotaxime and oxyimino-monobactam. The objective of the current review is to provide a better understanding of ESBL and the epidemiology of ESBL producing organisms which are among those responsible for antibiotic resistant strains. Globally, ESBLs are considered to be problematic, particularly in hospitalized patients. There is an increasing frequency of ESBL in different parts of the world. The high risk patients are those contaminated with ESBL producer strains as it renders treatment to be ineffective in these patients. Thus, there an immediate needs to identify EBSL and formulate strategic policy initiatives to reduce their prevalence.

  1. Kinetic studies on the inhibition of Proteus vulgaris beta-lactamase by imipenem.

    OpenAIRE

    Hashizume, T; Yamaguchi, A; Hirata, T; Sawai, T

    1984-01-01

    Imipenem was found to inhibit Proteus vulgaris beta-lactamase in a progressive manner. Kinetic experiments confirmed that the inactivated enzyme was not completely recovered after intact imipenem had been exhausted.

  2. Family disintegration: one fusarium verticillioides beta-lactamase at a time

    Science.gov (United States)

    Fusarium verticillioides is a mycotoxigenic fungus found commonly on maize, where it primarily exhibits asymptomatic endophytic growth. The F. verticillioides genome possesses approximately 30 regions that potentially encode beta-lactamase enzymatic domains. These enzymes are classically involved ...

  3. 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.

  4. Molecular identification of TEM-116 beta-lactamase gene in isolates ...

    African Journals Online (AJOL)

    Keywords: P. aeruginosa, Clinical isolates, Sequencing, TEM-116, Antibiotic susceptibility. Tropical Journal of ... organisms produce a wide range of beta ... TEM-1 is the most commonly encountered beta-lactamase in Gram-negative bacteria.

  5. Ertapenem susceptibility of extended spectrum beta-lactamase-producing organisms

    Directory of Open Access Journals (Sweden)

    Selby Edward B

    2007-06-01

    Full Text Available Abstract Background Infections caused by multiply drug resistant organisms such as extended spectrum beta-lactamase (ESBL-producing Escherichia coli and Klebsiella pneumoniae are increasing. Carbapenems (imipenem and meropenem are the antibiotics commonly used to treat these agents. There is limited clinical data regarding the efficacy of the newest carbapenem, ertapenem, against these organisms. Ertapenem susceptibility of ESBL-producing E. coli and K. pneumoniae clinical isolates were evaluated and compared to imipenem to determine if imipenem susceptibility could be used as a surrogate for ertapenem susceptibility. Methods 100 ESBL isolates (n = 34 E. coli and n = 66 K. pneumoniae collected from 2005–2006 clinical specimens at WRAMC were identified and tested for susceptibility by Vitek Legacy [bioMerieux, Durham, NC]. Ertapenem susceptibility was performed via epsilometer test (E-test [AB Biodisk, Solna, Sweden]. Results 100% of ESBL isolates tested were susceptible to ertapenem. 100% of the same isolates were also susceptible to imipenem. Conclusion These results, based on 100% susceptibility, suggest that ertapenem may be an alternative to other carbapenems for the treatment of infections caused by ESBL-producing E. coli and K. pneumoniae. Clinical outcomes studies are needed to determine if ertapenem is effective for the treatment of infection caused by these organisms. However, due to lack of resistant isolates, we are unable to conclude whether imipenem susceptibility accurately predicts ertapenem susceptibility.

  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. 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

  8. 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. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  9. The effect of a beta-lactamase inhibitor peptide on bacterial membrane structure and integrity: a comparative study.

    Science.gov (United States)

    Alaybeyoglu, Begum; Uluocak, Bilge Gedik; Akbulut, Berna Sariyar; Ozkirimli, Elif

    2017-05-01

    Co-administration of beta-lactam antibiotics and beta-lactamase inhibitors has been a favored treatment strategy against beta-lactamase-mediated bacterial antibiotic resistance, but the emergence of beta-lactamases resistant to current inhibitors necessitates the discovery of novel non-beta-lactam inhibitors. Peptides derived from the Ala46-Tyr51 region of the beta-lactamase inhibitor protein are considered as potent inhibitors of beta-lactamase; unfortunately, peptide delivery into the cell limits their potential. The properties of cell-penetrating peptides could guide the design of beta-lactamase inhibitory peptides. Here, our goal is to modify the peptide with the sequence RRGHYY that possesses beta-lactamase inhibitory activity under in vitro conditions. Inspired by the work on the cell-penetrating peptide pVEC, our approach involved the addition of the N-terminal hydrophobic residues, LLIIL, from pVEC to the inhibitor peptide to build a chimera. These residues have been reported to be critical in the uptake of pVEC. We tested the potential of RRGHYY and its chimeric derivative as a beta-lactamase inhibitory peptide on Escherichia coli cells and compared the results with the action of the antimicrobial peptide melittin, the beta-lactam antibiotic ampicillin, and the beta-lactamase inhibitor potassium clavulanate to get mechanistic details on their action. Our results show that the addition of LLIIL to the N-terminus of the beta-lactamase inhibitory peptide RRGHYY increases its membrane permeabilizing potential. Interestingly, the addition of this short stretch of hydrophobic residues also modified the inhibitory peptide such that it acquired antimicrobial property. We propose that addition of the hydrophobic LLIIL residues to the peptide N-terminus offers a promising strategy to design novel antimicrobial peptides in the battle against antibiotic resistance. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European

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

    International Nuclear Information System (INIS)

    Iqbal, F.I.; Farooqi, B.J.

    2012-01-01

    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)

  11. Extended-spectrum beta-lactamase-producing Enterobacteriaceae in hospital food: a risk assessment

    NARCIS (Netherlands)

    Stewardson, A.J.; Renzi, G.; Maury, N.; Vaudaux, C.; Brossier, C.; Fritsch, E.; Pittet, D.; Heck, M.; Zwaluw, K. van der; Reuland, E.A.; Laar, T. van; Snelders, E.; Vandenbroucke-Grauls, C.; Kluytmans, J.; Edder, P.; Schrenzel, J.; Harbarth, S.

    2014-01-01

    OBJECTIVE: Determine the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-PE) contamination of food and colonization of food handlers in a hospital kitchen and compare retrieved ESBL-PE strains with patient isolates. DESIGN: Cross-sectional study. SETTING: A

  12. Phenotypic and Genotypic Detection of Metallo-beta-lactamases among Imipenem-Resistant Gram Negative Isolates

    Directory of Open Access Journals (Sweden)

    Mohammad Mohammadzadeh

    2016-08-01

    Full Text Available Background:   Imipenem-resistant gram negative bacteria, resulting from metallo-beta-lactamase (MBLs-producing strains have been reported to be among the important causes of nosocomial infections and of serious therapeutic problem worldwide. Because of their broad range, potent carbapenemase activity and resistance to inhibitors, these enzymes can confer resistance to almost all beta-lactams. The prevalence of metallo-beta-lactamase among imipenem-resistant Acinetobacter spp., Pseudomonas spp. and Enerobacteriaceae isolates is determined.Methods:   In this descriptive study 864 clinical isolates of Acinetobacter spp., Pseudomonas spp. and Enterobacteriaceae, were initially tested for imipenem susceptibility. The metallo-beta-lactamase production was detected using combined disk diffusion, double disk synergy test, and Hodge test. Then all imipenem resistant isolates were tested by PCR for imp, vim and ndm genes. Results:   Among 864 isolates, 62 (7.17 % were imipenem-resistant. Positive phonetypic test for metallo-beta-lactamase was 40 (64.5%, of which 24 (17.1% and 16 (9.2% isolates were Acinetobacter spp. and Pseudomonas spp., respectively. By PCR method 30 (48.4% of imipenem resistant Acinetobacter, and Pseudomonas isolates were positive for MBL-producing genes. None of the Enterobacteriaceae isolates were positive for metallo-beta-lactamase activity. Conclusion:   The results of this study are indicative of the growing number of nosocomial infections associated with multidrug-resistant gram negative bacteria in this region leading to difficulties in antibiotic therapy. Thereby, using of phenotypic methods can be helpful for management of this problem.

  13. 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...... 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...

  14. 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.

  15. SME-3, a novel member of the Serratia marcescens SME family of carbapenem-hydrolyzing beta-lactamases.

    Science.gov (United States)

    Queenan, Anne Marie; Shang, Wenchi; Schreckenberger, Paul; Lolans, Karen; Bush, Karen; Quinn, John

    2006-10-01

    Imipenem-resistant Serratia marcescens isolates were cultured from a lung transplant patient given multiple antibiotics over several months. The strains expressed SME-3, a beta-lactamase of the rare SME carbapenem-hydrolyzing family. SME-3 differed from SME-1 by a single amino acid substitution of tyrosine for histidine at position 105, but the two beta-lactamases displayed similar hydrolytic profiles.

  16. [Extended-spectrum beta-lactamase detection in Enterobacteriaceae and antibiotic susceptibility analysis].

    Science.gov (United States)

    Cao, Wei; Tong, Ming-hua; Wang, Ji-gui

    2002-02-28

    To detect the extended-spectrum beta-lactamases (ESBLs) in family Enterobacteriaceae and analyze the antibiotic susceptibility of those ESBLs-producing strains. ESBLs were determined by the double-disk confirmatory test and 8 antibiotic susceptibilities were tested with the disk disffusion method in those strains producing ESBLs. Forty-seven ESBLs-producing strains comprised of 25 of E. coli, 14 of K. pneumoniae, 5 of E. cloacae, 1 of K. oxytoca, 1 of K. rhinoscleromatis, and 1 of S. liquefaciens. The susceptibility rates of those strains were: 100% for imipenem and meropenem, 89.4% for piperacillin/tazobactam, 72.4% for cefoxitin and 65.9% for cefotetan. E. coli and K. pneumoniae are the prime strains producing ESBLs in Enterobacteriaceae. Imipenem and meropenem are the best drugs to deal with those ESBLs-producing strains. Piperacillin/tazobactam is better than cephamycins and other beta-lactama/beta-lactamase inhibitor combination.

  17. Amino acid residues that contribute to substrate specificity of class A beta-lactamase SME-1.

    Science.gov (United States)

    Majiduddin, Fahd K; Palzkill, Timothy

    2005-08-01

    Carbapenem antibiotics are used as antibiotics of last resort because they possess a broad spectrum of antimicrobial activity and are not easily hydrolyzed by beta-lactamases. Recently, class A enzymes, such as the SME-1, NMC-A, and IMI-1 beta-lactamases, have been identified with the capacity to hydrolyze carbapenem antibiotics. Traditional class A beta-lactamases, such as TEM-1 and SHV-1, are unable to hydrolyze carbapenem antibiotics and exhibit some differences in sequence from those that are able to hydrolyze carbapenem antibiotics. The positions that differ may contribute to the unique substrate specificity of the class A carbapenemase SME-1. Codons in the SME-1 gene representing residues 104, 105, 132, 167, 237, and 241 were randomized by site-directed mutagenesis, and functional mutants were selected for the ability to hydrolyze imipenem, ampicillin, or cefotaxime. Although several positions are important for hydrolysis of beta-lactam antibiotics, no single position was found to uniquely contribute to carbapenem hydrolysis. The results of this study support a model whereby the carbapenemase activity of SME-1 is due to a highly distributed set of interactions that subtly alter the structure of the active-site pocket.

  18. A fitness cost associated with the antibiotic resistance enzyme SME-1 beta-lactamase.

    Science.gov (United States)

    Marciano, David C; Karkouti, Omid Y; Palzkill, Timothy

    2007-08-01

    The bla(TEM-1) beta-lactamase gene has become widespread due to the selective pressure of beta-lactam use and its stable maintenance on transferable DNA elements. In contrast, bla(SME-1) is rarely isolated and is confined to the chromosome of carbapenem-resistant Serratia marcescens strains. Dissemination of bla(SME-1) via transfer to a mobile DNA element could hinder the use of carbapenems. In this study, bla(SME-1) was determined to impart a fitness cost upon Escherichia coli in multiple genetic contexts and assays. Genetic screens and designed SME-1 mutants were utilized to identify the source of this fitness cost. These experiments established that the SME-1 protein was required for the fitness cost but also that the enzyme activity of SME-1 was not associated with the fitness cost. The genetic screens suggested that the SME-1 signal sequence was involved in the fitness cost. Consistent with these findings, exchange of the SME-1 signal sequence for the TEM-1 signal sequence alleviated the fitness cost while replacing the TEM-1 signal sequence with the SME-1 signal sequence imparted a fitness cost to TEM-1 beta-lactamase. Taken together, these results suggest that fitness costs associated with some beta-lactamases may limit their dissemination.

  19. Cloning and sequence of the gene encoding a cefotaxime-hydrolyzing class A beta-lactamase isolated from Escherichia coli.

    Science.gov (United States)

    Ishii, Y; Ohno, A; Taguchi, H; Imajo, S; Ishiguro, M; Matsuzawa, H

    1995-01-01

    Escherichia coli TUH12191, which is resistant to piperacillin, cefazolin, cefotiam, ceftizoxime, cefuzonam, and aztreonam but is susceptible to cefoxitin, latamoxef, flomoxef, and imipenem, was isolated from the urine of a patient treated with beta-lactam antibiotics. The beta-lactamase (Toho-1) purified from the bacteria had a pI of 7.8, had a molecular weight of about 29,000, and hydrolyzed beta-lactam antibiotics such as penicillin G, ampicillin, oxacillin, carbenicillin, piperacillin, cephalothin, cefoxitin, cefotaxime, ceftazidime, and aztreonam. Toho-1 was markedly inhibited by beta-lactamase inhibitors such as clavulanic acid and tazobactam. Resistance to beta-lactams, streptomycin, spectinomycin, sulfamethoxazole, and trimethoprim was transferred by conjugational transfer from E. coli TUH12191 to E. coli ML4903, and the transferred plasmid was about 58 kbp, belonging to incompatibility group M. The cefotaxime resistance gene for Toho-1 was subcloned from the 58-kbp plasmid by transformation of E. coli MV1184. The sequence of the gene for Toho-1 was determined, and the open reading frame of the gene consisted of 873 or 876 bases (initial sequence, ATGATG). The nucleotide sequence of the gene (DDBJ accession number D37830) was found to be about 73% homologous to the sequence of the gene encoding a class A beta-lactamase produced by Klebsiella oxytoca E23004. According to the amino acid sequence deduced from the DNA sequence, the precursor consisted of 290 or 291 amino acid residues, which contained amino acid motifs common to class A beta-lactamases (70SXXK, 130SDN, and 234KTG). Toho-1 was about 83% homologous to the beta-lactamase mediated by the chromosome of K. oxytoca D488 and the beta-lactamase mediated by the plasmid of E. coli MEN-1. Therefore, the newly isolated beta-lactamase Toho-1 produced by E. coli TUH12191 is similar to beta-lactamases produced by K. oxytoca D488, K. oxytoca E23004, and E. coli MEN-1 rather than to mutants of TEM or SHV enzymes

  20. 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...... were tested towards 21 antimicrobial agents. PCR and sequencing were used to determine the underlying genetic determinants responsible for the ESBL phenotypes. The transferability of the ESBL phenotypes was tested by conjugation to a susceptible Salmonella enterica serovar Dublin and plasmid....... Finally, the bla(ACC-1) gene was cloned from a S. Bareilly isolate and was found to be present on indistinguishable plasmids in all S. Bareilly isolates examined as well as in a S. Braenderup isolate and a S. Infantis isolate. Conclusions: Our data underscore the diversity of ESBL genes in Salmonella...

  1. 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.

  2. Varying occurrence of extended-spectrum beta-lactamase bacteria among three produce types

    KAUST Repository

    Toh, Benjamin E. W.; Bokhari, Osama Mohammed; Kutbi, Abdullah; Haroon, Mohamed; Mantilla-Calderon, David; Zowawi, Hosam; Hong, Pei-Ying

    2017-01-01

    Three types of vegetables were sampled and evaluated over 1.5 years to determine differences in their associated bacterial isolates. Particular emphasis was placed on identifying pathogenic strains that were positive for extended spectrum beta-lactamase (ESBL). Quantitative estimates of the microbial risk associated with the ESBL-positive pathogens showed that different produce types may incur varying levels of ingestion risk. Most of the currently reported ESBL-positive bacterial isolates have been identified in nosocomial environments. However, the carriage of such drug-resistant bacteria in vegetables suggests a possible connection between our daily diet and human health.

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

    Science.gov (United States)

    Rasmussen, Mette Marie; Opintan, Japheth A; 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 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 are potential

  4. 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.

  5. 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

  6. Cross-class resistance to non-beta-lactam antimicrobials in extended-spectrum beta-lactamase-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Procop, Gary W; Tuohy, Marion J; Wilson, Deborah A; Williams, Delisa; Hadziyannis, Emilia; Hall, Gerri S

    2003-08-01

    Extended spectrum beta-lactamases are modified beta-lactamase enzymes that impart resistance to third-generation cephalosporins and make all beta-lactam antibiotics and cephalosporins useless for therapy. We compared the antimicrobial susceptibility profiles of extended-spectrum beta-lactamase (ESBL)-producing and non-ESBL-producing isolates of Klebsiella pneumoniae. The ESBL producers had significantly diminished susceptibility compared with the non-ESBL producers for gentamicin (P < .001), tobramycin (P < .001), amikacin (P < .005), trimethoprim-sulfamethoxazole (P < .01), ciprofloxacin (P < .001), and nitrofurantoin (P < .001). All isolates were susceptible to imipenem. ESBL-producing K pneumoniae may also be resistant to non-beta-lactam antibiotics. Therefore, susceptibility testing of these isolates is critical for guiding therapy.

  7. 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

    the role of beta-lactamase in the pharmacokinetics (PK) and pharmacodynamics (PD) of ceftazidime and imipenem on P. aeruginosa biofilms. P. aeruginosa PAO1 and its corresponding beta-lactamase-overproducing mutant, PA Delta DDh2Dh3, were used in this study. Biofilms of these two strains in flow chambers......, 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...... 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...

  8. Raw pet food as a risk factor for shedding of extended-spectrum beta-lactamase-producing Enterobacteriaceae in household cats

    NARCIS (Netherlands)

    Baede, V.O. (Valérie O.); E.M. Broens; Spaninks, M.P. (Mirlin P.); Timmerman, A.J. (Arjen J.); Graveland, H. (Haitske); J.A. Wagenaar (Jaap); B. Duim; Hordijk, J. (Joost)

    2017-01-01

    textabstractBackground: Close contact between pets and owners provides the opportunity for transmission of antimicrobial resistant organisms like extended-spectrum beta-lactamase (ESBL)/AmpC beta-lactamase (AmpC)-producing Enterobacteriaceae, posing a risk to public health.

  9. Raw pet food as a risk factor for shedding of extended-spectrum beta-lactamase-producing Enterobacteriaceae in household cats

    NARCIS (Netherlands)

    Baede, Valérie O; Broens, Els M|info:eu-repo/dai/nl/314627723; Spaninks, Mirlin P; Timmerman, Arjen J; Graveland, Haitske|info:eu-repo/dai/nl/304841838; Wagenaar, Jaap A|info:eu-repo/dai/nl/126613354; Duim, Birgitta|info:eu-repo/dai/nl/143855352; Hordijk, Joost|info:eu-repo/dai/nl/314839542

    2017-01-01

    BACKGROUND: Close contact between pets and owners provides the opportunity for transmission of antimicrobial resistant organisms like extended-spectrum beta-lactamase (ESBL)/AmpC beta-lactamase (AmpC)-producing Enterobacteriaceae, posing a risk to public health. OBJECTIVES: To investigate whether

  10. Raw pet food as a risk factor for shedding of extended-spectrum beta-lactamase-producing Enterobacteriaceae in household cats

    NARCIS (Netherlands)

    Baede, Valérie O.; Broens, Els M.; Spaninks, Mirlin P.; Timmerman, Arjen J.; Graveland, Haitske; Wagenaar, Jaap A.; Duim, Birgitta; Hordijk, Joost

    2017-01-01

    Background: Close contact between pets and owners provides the opportunity for transmission of antimicrobial resistant organisms like extended-spectrum beta-lactamase (ESBL)/AmpC beta-lactamase (AmpC)-producing Enterobacteriaceae, posing a risk to public health. Objectives: To investigate whether

  11. Chromosome-encoded narrow-spectrum Ambler class A beta-lactamase GIL-1 from Citrobacter gillenii.

    Science.gov (United States)

    Naas, Thierry; Aubert, Daniel; Ozcan, Ayla; Nordmann, Patrice

    2007-04-01

    A novel beta-lactamase gene was cloned from the whole-cell DNA of an enterobacterial Citrobacter gillenii reference strain that displayed a weak narrow-spectrum beta-lactam-resistant phenotype and was expressed in Escherichia coli. It encoded a clavulanic acid-inhibited Ambler class A beta-lactamase, GIL-1, with a pI value of 7.5 and a molecular mass of ca. 29 kDa. GIL-1 had the highest percent amino acid sequence identity with TEM-1 and SHV-1, 77%, and 67%, respectively, and only 46%, 31%, and 32% amino acid sequence identity with CKO-1 (C. koseri), CdiA1 (C. diversus), and SED-1 (C. sedlaki), respectively. The substrate profile of the purified GIL-1 was similar to that of beta-lactamases TEM-1 and SHV-1. The blaGIL-1 gene was chromosomally located, as revealed by I-CeuI experiments, and was constitutively expressed at a low level in C. gillenii. No gene homologous to the regulatory ampR genes of chromosomal class C beta-lactamases was found upstream of the blaGIL-1 gene, which fits the noninducibility of beta-lactamase expression in C. gillenii. Rapid amplification of DNA 5' ends analysis of the promoter region revealed putative promoter sequences that diverge from what has been identified as the consensus sequence in E. coli. The blaGIL-1 gene was part of a 5.5-kb DNA fragment bracketed by a 9-bp duplication and inserted between the d-lactate dehydrogenase gene and the ydbH genes; this DNA fragment was absent in other Citrobacter species. This work further illustrates the heterogeneity of beta-lactamases in Citrobacter spp., which may indicate that the variability of Citrobacter species is greater than expected.

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

    International Nuclear Information System (INIS)

    Roshan, M.

    2011-01-01

    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)

  13. Method for phenotypic detection of extended-spectrum beta-lactamases in enterobacter species in the routine clinical setting

    NARCIS (Netherlands)

    Stuart, J.C.; Diederen, B.; Al Naiemi, N.; Fluit, A.; Arents, N.; Thijsen, S.; Vlaminckx, B.; Mouton, J.W.; Leverstein-van Hall, M.

    2011-01-01

    In 271 Enterobacter blood culture isolates from 12 hospitals, extended-spectrum beta-lactamase (ESBL) prevalence varied between 0% and 30% per hospital. High prevalence was associated with dissemination, indicating the potential relevance of infection control measures. Screening with cefepime or

  14. 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

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

    International Nuclear Information System (INIS)

    Hassan, A.; Usman, J.; Kalim, F.; Gill, M.M.; Khalid, A.; Iqbal, M.; Ingram, P.

    2013-01-01

    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)

  16. 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

    -level of 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 stable...

  17. 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.

    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

  18. In vitro effects of beta-lactams combined with beta-lactamase inhibitors against methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Kobayashi, S; Arai, S; Hayashi, S; Sakaguchi, T

    1989-01-01

    The effects of combinations of beta-lactams with two beta-lactamase inhibitors, sulbactam and clavulanic acid, were determined in vitro against 22 clinical isolates of methicillin-resistant Staphylococcus aureus. Combinations of cefpirome, cefotaxime, and cefazolin with sulbactam (10 micrograms/ml) showed synergistic effects against more than 70% of the strains. Combinations of methicillin and penicillin G with sulbactam also showed synergistic effects against 50 and 68% of the strains, respectively, while cefotiam, moxalactam, flomoxef, and cefmetazole in combination with sulbactam showed such effects against only 40% or fewer. Clavulanic acid was synergistic only when combined with penicillin G, the effect probably being due to the beta-lactamase inhibition by the inhibitor. Sulbactam did not improve the antimicrobial activities of the beta-lactams against methicillin-susceptible S. aureus strains. At 42 degrees C the MICs of cefotaxime, methicillin, and flomoxef alone were markedly decreased from the values at 35 degrees C, and no synergy between these beta-lactams and sulbactam appeared. The resistance to penicillin G was not inhibited by incubation at 42 degrees C, and combinations of penicillin G with sulbactam and clavulanic acid showed synergy. The amounts of beta-lactamase produced were not related to the decreases in the MICs of the beta-lactams, except for penicillin G combined with sulbactam. Clavulanic acid showed slightly stronger beta-lactamase-inhibiting activity than sulbactam did. These results suggest that the synergy between sulbactam and the beta-lactams, except for penicillin G, may not be due to beta-lactamase inhibition but to suppression of the methicillin-resistant S. aureus-specific resistance based on other factors. PMID:2786369

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

    Science.gov (United States)

    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 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

  20. Substrate-induced inactivation of the OXA2 beta-lactamase.

    Science.gov (United States)

    Ledent, P; Frère, J M

    1993-01-01

    The hydrolysis time courses of 22 beta-lactam antibiotics by the class D OXA2 beta-lactamase were studied. Among these, only three appeared to correspond to the integrated Henri-Michaelis equation. 'Burst' kinetics, implying branched pathways, were observed with most penicillins, cephalosporins and with flomoxef and imipenem. Kinetic parameters characteristic of the different phases of the hydrolysis were determined for some substrates. Mechanisms generally accepted to explain such reversible partial inactivations involving branches at either the free enzyme or the acyl-enzyme were inadequate to explain the enzyme behaviour. The hydrolysis of imipenem was characterized by the occurrence of two 'bursts', and that of nitrocefin by a partial substrate-induced inactivation complicated by a competitive inhibition by the hydrolysis product. PMID:8240304

  1. 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

    2015-01-01

    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...... 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......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...

  2. Varying occurrence of extended-spectrum beta-lactamase bacteria among three produce types

    KAUST Repository

    Toh, Benjamin E. W.

    2017-07-07

    A monitoring effort that spanned across 1.5 years was conducted to examine three types of produce-associated microbiota. The average amount of antibiotic-resistant bacteria recovered from lettuce, tomato, and cucumber was 1.02 × 1010, 2.05 × 107, and 4.78 × 109 cells per 50 g of each produce, respectively. A total of 480 bacterial isolates were obtained and identified from their 16S rRNA genes, revealing isolates that were ubiquitously recovered from all three types of produce. However, sporadic presence of Klebsiella pneumoniae and Acinetobacter baumannii was detected on lettuce and cucumbers but not tomatoes. End-point PCR revealed that the K. pneumoniae and A. baumannii isolates were positive for genes encoding extended spectrum beta-lactamase. Whole genome sequencing of two of the K. pneumoniae isolates further suggested the presence of the blaCTX-M-15 gene in a conjugative plasmid, as well as other antibiotic resistance genes and virulence-associated traits in either conjugative plasmids or the chromosomal genome. Quantitative microbial risk assessment indicated varying levels of ingestion risk associated with different types of produce. In particular, the risk arising from ESBL-positive K. pneumoniae in lettuce, but not in cucumbers or tomatoes, was higher than the acceptable annual risk of 10−4. Practical applications Three types of vegetables were sampled and evaluated over 1.5 years to determine differences in their associated bacterial isolates. Particular emphasis was placed on identifying pathogenic strains that were positive for extended spectrum beta-lactamase (ESBL). Quantitative estimates of the microbial risk associated with the ESBL-positive pathogens showed that different produce types may incur varying levels of ingestion risk. Most of the currently reported ESBL-positive bacterial isolates have been identified in nosocomial environments. However, the carriage of such drug-resistant bacteria in vegetables suggests a possible connection

  3. 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 development of resistance to beta-lactam antibiotics and the occurrence of high beta-lactamase producing strains and between the MIC of the beta-lactams and the levels of beta-lactamase expression. Partially derepressed mutants, characterized by high basal levels of beta-lactamase with the possibility...... of 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 more susceptible to antibiotics and produce less beta-lactamase than the non-mucoid paired isolates. We propose that the non-mucoid isolates are exposed to a relatively higher antibiotic pressure than the mucoid isolates and therefore, they become easily antibiotic resistant and in consequence produce...

  4. ICU Acquisition Rate, Risk Factors, and Clinical Significance of Digestive Tract Colonization With Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Detsis, Marios; Karanika, Styliani; Mylonakis, Eleftherios

    2017-04-01

    To evaluate the acquisition rate, identify risk factors, and estimate the risk for subsequent infection, associated with the colonization of the digestive tract with extended-spectrum beta-lactamase-producing Enterobacteriaceae during ICU-hospitalization. PubMed, EMBASE, and reference lists of all eligible articles. Included studies provided data on ICU-acquired colonization with extended-spectrum beta-lactamase-producing Enterobacteriaceae in previously noncolonized and noninfected patients and used the double disk synergy test for extended-spectrum beta-lactamase-producing Enterobacteriaceae phenotypic confirmation. Studies reporting extended-spectrum beta-lactamase-producing Enterobacteriaceae outbreaks or data on pediatric population were excluded. Two authors independently assessed study eligibility and performed data extraction. Thirteen studies (with 15,045 ICUs-patients) were evaluated using a random-effect model and a meta-regression analysis. The acquisition rate of digestive tract colonization during ICU stay was 7% (95% CI, 5-10) and it varies from 3% (95% CI, 2-4) and 4% (95% CI, 2-6) in the Americas and Europe to 21% (95% CI, 9-35) in the Western Pacific region. Previous hospitalization (risk ratio, 1.57 [95% CI, 1.07-2.31]) or antibiotic use (risk ratio, 1.65 [95% CI, 1.15-2.37]) and exposure to beta-lactams/beta-lactamase inhibitors (risk ratio, 1.78 [95% CI, 1.24-2.56]) and carbapenems (risk ratio, 2.13 [95% CI, 1.49-3.06]) during the ICU stay were independent risk factors for ICU-acquired colonization. Importantly, colonized patients were more likely to develop an extended-spectrum beta-lactamase-producing Enterobacteriaceae infection (risk ratio, 49.62 [95% CI, 20.42-120.58]). The sensitivity and specificity of prior colonization to predict subsequent extended-spectrum beta-lactamase-producing Enterobacteriaceae infection were 95.1% (95% CI, 54.7-99.7) and 89.2% (95% CI, 77.2-95.3), respectively. The ICU acquisition rate of extended-spectrum beta-lactamase

  5. Seagulls of the Berlengas natural reserve of Portugal as carriers of fecal Escherichia coli harboring CTX-M and TEM extended-spectrum beta-lactamases.

    Science.gov (United States)

    Poeta, Patricia; Radhouani, Hajer; Igrejas, Gilberto; Gonçalves, Alexandre; Carvalho, Carlos; Rodrigues, Jorge; Vinué, Laura; Somalo, Sergio; Torres, Carmen

    2008-12-01

    Escherichia coli isolates containing the following extended-spectrum beta-lactamases have been detected in 11 of 57 fecal samples (19.3%) in Berlengas Island seagulls: TEM-52 (eight isolates), CTX-M-1 (one isolate), CTX-M-14a (one isolate), and CTX-M-32 (one isolate). Most of the extended-spectrum beta-lactamase-positive isolates harbored class 1 or class 2 integrons, which included different antibiotic resistance gene cassettes.

  6. Burden of different beta-lactamase classes among clinical isolates of AmpC-producing Pseudomonas aeruginosa in burn patients: A prospective study

    OpenAIRE

    Kumar, V.; Sen, M. R.; Nigam, C.; Gahlot, R.; Kumari, S.

    2012-01-01

    Background: Pseudomonas aeruginosa is one of the most common pathogens causing infections in burns, and shows increasing resistance to β-lactam antibiotics by producing different classes of beta-lactamases. It is also not unusual to find a single isolate that expresses multiple β-lactamase enzymes, further complicating the treatment options. Thus, in this study, we aimed to determine the coexistence of different beta-lactamase enzymes in clinical isolates of P. aeruginosa in the burn ward. Ma...

  7. Method for Phenotypic Detection of Extended-Spectrum Beta-Lactamases in Enterobacter Species in the Routine Clinical Setting ▿

    Science.gov (United States)

    Stuart, James Cohen; Diederen, Bram; al Naiemi, Nashwan; Fluit, Ad; Arents, Niek; Thijsen, Steven; Vlaminckx, Bart; Mouton, Johan W.; Leverstein-van Hall, Maurine

    2011-01-01

    In 271 Enterobacter blood culture isolates from 12 hospitals, extended-spectrum beta-lactamase (ESBL) prevalence varied between 0% and 30% per hospital. High prevalence was associated with dissemination, indicating the potential relevance of infection control measures. Screening with cefepime or Vitek 2, followed by a cefepime/cefepime-clavulanate Etest, was an accurate strategy for ESBL detection in Enterobacter isolates (positive predictive value, 100%; negative predictive value, 99%). PMID:21562100

  8. The 1.4 Å Crystal Structure of the Class D [beta]-Lactamase OXA-1 Complexed with Doripenem

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, Kyle D.; Karpen, Mary E.; Bonomo, Robert A.; Leonard, David A.; Powers, Rachel A.; (Grand Valley); (Case Western U.-Med)

    2010-01-12

    The clinical efficacy of carbapenem antibiotics depends on their resistance to the hydrolytic action of {beta}-lactamase enzymes. The structure of the class D {beta}-lactamase OXA-1 as an acyl complex with the carbapenem doripenem was determined to 1.4 {angstrom} resolution. Unlike most class A and class C carbapenem complexes, the acyl carbonyl oxygen in the OXA-1-doripenem complex is bound in the oxyanion hole. Interestingly, no water molecules were observed in the vicinity of the acyl linkage, providing an explanation for why carbapenems inhibit OXA-1. The side chain amine of K70 remains fully carboxylated in the acyl structure, and the resulting carbamate group forms a hydrogen bond to the alcohol of the 6{alpha}-hydroxyethyl moiety of doripenem. The carboxylate attached to the {beta}-lactam ring of doripenem is stabilized by a salt bridge to K212 and a hydrogen bond with T213, in lieu of the interaction with an arginine side chain found in most other {beta}-lactamase-{beta}-lactam complexes (e.g., R244 in the class A member TEM-1). This novel set of interactions with the carboxylate results in a major shift of the carbapenem's pyrroline ring compared to the structure of the same ring in meropenem bound to OXA-13. Additionally, bond angles of the pyrroline ring suggest that after acylation, doripenem adopts the {Delta}{sup 1} tautomer. These findings provide important insights into the role that carbapenems may have in the inactivation process of class D {beta}-lactamases.

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

    International Nuclear Information System (INIS)

    El-Khizzi, Noura A.; Bakheshwain, S. M.

    2006-01-01

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

    Science.gov (United States)

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

    1983-08-01

    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 observed for HMS-1 and PSE-1 beta-lactamases. The HMS-1 sample produced two highly resolvable enzyme bands in agarose gels rather than the single faint enzyme band observed on polyacrylamide gels. The PSE-1 sample showed an isoelectric point shift of 0.2 pH unit between polyacrylamide and agarose gel (pI 5.7 and 5.5, respectively). The short focusing time, lack of toxic hazard, and ease of formulation make agarose a practical medium for the characterization of beta-lactamases.

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

    International Nuclear Information System (INIS)

    Gill, M. M.; Usman, J.; Hassan, A.; Kaleem, F.; Anjum, R.

    2015-01-01

    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)

  12. 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.

  13. Predicament in detection and reporting of extended spectrum beta lactamase production in routine antibiotic susceptibility testing

    International Nuclear Information System (INIS)

    Butt, T.; Butt, E.; Raza, S.

    2017-01-01

    This descriptive and cross-sectional study was planned to determine the dilemma of inadvertent detection of extended spectrum beta lactamase (ESBL) production in Enterobacteriaceaewhen using inhibition zone size of antibiotic disks of Cefotaxime or Aztreonam in routine antibiotic susceptibility testing as recommended by Clinical Laboratory Standards Institute (CLSI). Screening and double disk tests were adopted as per CLSI. Escherichia coli ATCC 25922 was used as control strain. Among total specimens of 5346, there were 348 isolates of Escherichia coli(n=235), Klebsiella pneumonia (n=92), Klebsiella oxytoca(n=3) or Proteus mirabilus(n=18). The screening method recommended by CLSI significantly falsely detected ESBL production in 79 (32.3%) isolates (p<0.0001). ESBL detection is important as its frequency is high and treatment of the infection varies with the presence and absence of ESBL. To avoid false reporting, proper phenotypic detection of ESBL confirmatory method-like double-disk synergy test, should be used routinely. (author)

  14. 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

  15. Nitrofurantoin and Fosfomycin for Extended Spectrum Beta-lactamases Producing Escherichia coli and Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    Neeraj Kumar Tulara

    2018-01-01

    Full Text Available Urinary tract infection (UTI is a common and painful human illness that, unfortunately not responsive to commonly used antibiotics in current practice. The role of fosfomycin and nitrofurantoin in the era of growing bacteria resistance has been widely discussed. In this study, we aimed to know the local antimicrobial susceptibilities, fosfomycin and nitrofurantoin susceptibility in particular, for urinary extended-spectrum-beta-lactamase-producing Escherichia coli and Escherichia pneumoniae (ESBL-EC and ESBL-KP isolates in our hospital. We collected 464 urine isolates, including 384 ESBL-EC and 80 ESBL-KP isolates. Of 464 urine isolates culture positive ESBL-UTIs, EC caused 384 (82.75%, followed by Klebsiella in 80 (17.24%. Carbapenems and Colistin seems to remain as the first line therapy for the majority of ESBL-UTIs in the local setting. Colistin and fosfomycin remains the most sensitive antibiotic while nitrofurantoin still preserves the good sensitivity against ESBL and found to be an only oral sensitive antibiotic.

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

    International Nuclear Information System (INIS)

    Khan, I.U.; Mirza, I.A.; Ali, S.; Hussain, A.

    2014-01-01

    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)

  17. Covalent docking of selected boron-based serine beta-lactamase inhibitors

    Science.gov (United States)

    Sgrignani, Jacopo; Novati, Beatrice; Colombo, Giorgio; Grazioso, Giovanni

    2015-05-01

    AmpC β-lactamase is a hydrolytic enzyme conferring resistance to β-lactam antibiotics in multiple Gram-negative bacteria. Therefore, identification of non-β-lactam compounds able to inhibit the enzyme is crucial for the development of novel antibacterial therapies. In general, AmpC inhibitors have to engage the highly solvent-exposed catalytic site of the enzyme. Therefore, understanding the implications of ligand-protein induced-fit and water-mediated interactions behind the inhibitor-enzyme recognition process is fundamental for undertaking structure-based drug design process. Here, we focus on boronic acids, a promising class of beta-lactamase covalent inhibitors. First, we optimized a docking protocol able to reproduce the experimentally determined binding mode of AmpC inhibitors bearing a boronic group. This goal was pursued (1) performing rigid and flexible docking calculations aiming to establish the role of the side chain conformations; and (2) investigating the role of specific water molecules in shaping the enzyme active site and mediating ligand protein interactions. Our calculations showed that some water molecules, conserved in the majority of the considered X-ray structures, are needed to correctly predict the binding pose of known covalent AmpC inhibitors. On this basis, we formalized our findings in a docking and scoring protocol that could be useful for the structure-based design of new boronic acid AmpC inhibitors.

  18. Extended spectrum beta lactamase (ESBL) producing bacteria urinary tract infections and complex pediatric urology.

    Science.gov (United States)

    Wragg, Ruth; Harris, Anna; Patel, Mitul; Robb, Andrew; Chandran, Harish; McCarthy, Liam

    2017-02-01

    Extended spectrum beta lactamase (ESBL) producing bacteria are resistant to most beta-lactam antibiotics including third-generation cephalosporins, quinolones and aminoglycosides. This resistance is plasmid-borne and can spread between species. Management of ESBL is challenging in children with recurrent urinary tract infections (UTIs) and complex urological abnormalities. We aim to quantify the risk in children and specifically in urological patients. Retrospective review of a microbiology database (April 2014 to November 2015). This identified urine isolates, pyuria, ESBL growth and patient demographics. Data analysis was by Chi square, Mann-Whitney U-test and ANOVA. A P value of 10×10 6 WC/L). 136 urine cultures (n=79 patients) grew purely ESBL. Overall, 5.2% of urine isolates were ESBL and 9.5% isolates with pyuria (>100×10 6 WC/L) had ESBL, whereas only 22/1032 (2.1%) with no pyuria, (Pantibiotics). Over the study period, there was no significant rise of the monthly incidence between 2014 and 2015 (ANOVA P=0.1). This study is the first to document the incidence of ESBL in children (5%), and estimate the frequency of possible plasmid transmission between bacterial species in children. This quantifies the risk of ESBL, especially to urology patients, and mandates better antibiotic stewardship. Level IIc. Copyright © 2017. Published by Elsevier Inc.

  19. PEDIATRIC URINARY INFECTIONS, CAUSED BY EXTENDED-SPECTRUM BETA-LACTAMASE - PRODUCING MICROORGANISMS IN VARNA, BULGARIA

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    Neli M. Ermenlieva

    2016-05-01

    Full Text Available Background: Extended-spectrum beta-lactamase (ESBLs producing bacteria are microorganisms which have the ability to hydrolyze β-lactame ring of a large part of the antibiotics, commonly used to treat bacterial infections including urinary tract infections. Purpose: The aim of this study is present the epidemiology of childhood urinary tract infections caused by ESBL-producing strains in Varna, Bulgaria. Material/methods: A total of 3895 urine samples of children patients (aged 0 to 18 years were examined during the period 2010-2012 for presence of ESBL-producing bacteria. Results: Six percent of the tested urinary samples were positive for ESBL production. All of the isolates were resistant to ampicillin, piperacillin, cephalothin, cefprozil, cefuroxime, ceftriaxone, ceftazidime, levofloxacin, cefaclor, but were were sensitive to meropenem and imipenem. Conclusions: Cephalosporins and penicillins are the most used antibiotics in Bulgaria, but they should be very precisely prescribed in medical practice, because otherwise preconditions for maintaining high share of ESBLs are created.

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

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    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.

  1. Assessment of AmpC Beta-Lactamase Genes among Clinical Escherichia coli Isolates

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    HedrooshaMolla Agha-Mirzaeie

    2015-11-01

    Full Text Available Background: AmpC bta lactamases play a significant role in creating resistance to third generation cephalosporins worldwide. They mostly express on chromosome of Enterobacteriaceae especially Escherichia coli and cause consequential problem inclinical treatment and lead to failure in diagnosis and phenotypic test recommended byClinical and Laboratory Standards Institute.Methods:Totally 200 E. coli isolates from different hospitals of Tehran were collected. The isolates were screened by disk diffusion method according to the CLSI guidelines. The profiles and prevalence surveys of AmpC (Dha, CITM, Mox and FOX-type β-lactamase genes in clinical isolates of E. coli by phenotypic and molecular methods.  Results:Out of 200 Ecoli isolated, 115 (89.8% and 13 (10.2% isolates were identified as ESBL- and AmpC- beta-lactamase producers, respectively. Among mpC producers, 13 (100% and 5 (38.5% isolates was reported by PCR assay as bla-CITM and Dha respectively. Mox and FOX genes were not detected in any sample.Conclusions:Our results highlight the importance of using molecular detection methods to identify β-lactamase-producer that have resistance to antibiotics. 

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

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    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.

  3. Characteristics of bacteremia caused by extended-spectrum beta-lactamase-producing Proteus mirabilis.

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    Kurihara, Yoko; Hitomi, Shigemi; Oishi, Tsuyoshi; Kondo, Tsukasa; Ebihara, Tsugio; Funayama, Yasunori; Kawakami, Yasushi

    2013-10-01

    Although Proteus mirabilis is a common human pathogen, bacteremia caused by the organism, especially strains producing extended-spectrum beta-lactamase (ESBL), has rarely been investigated. We examined 64 cases of P. mirabilis bacteremia identified in the Minami Ibaraki Area, Japan, between 2001 and 2010 and compared the characteristics of cases with ESBL-producing and ESBL-non-producing strains (13 and 51 cases, respectively). All ESBL-producing strains with the gene encoding the CTX-M-2-group were genetically nonidentical. Isolation of ESBL-producing strains was significantly associated with onset in a hospital (p = 0.030), receiving hemodialysis (p = 0.0050), and previous antibiotic use within 1 month (p = 0.036; especially penicillin and/or cephalosporin (p = 0.010) and fluoroquinolone (p = 0.0069)). Isolation was also associated with inappropriate antibiotic therapy on the 1st and 4th days (p = 0.011 and 0.032, respectively) but not with mortality on the 30th day. These findings indicate that, for P. mirabilis bacteremia, isolation of ESBL-producing strains causes delay of initiating appropriate antimicrobial therapy but may not be associated with mortality.

  4. Overcoming resistance to beta-lactamase inhibitors: comparing sulbactam to novel inhibitors against clavulanate resistant SHV enzymes with substitutions at Ambler position 244.

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    Thomson, Jodi M; Distler, Anne M; Bonomo, Robert A

    2007-10-09

    Amino acid changes at Ambler position R244 in class A TEM and SHV beta-lactamases confer resistance to ampicillin/clavulanate, a beta-lactam/beta-lactamase inhibitor combination used to treat serious infections. To gain a deeper understanding of this resistance phenotype, we investigated the activities of sulbactam and two novel penem beta-lactamase inhibitors with sp2 hybridized C3 carboxylates and bicyclic R1 side chains against a library of SHV beta-lactamase variants at the 244 position. Compared to SHV-1 expressed in Escherichia coli, all 19 R244 variants exhibited increased susceptibility to ampicillin/sulbactam, an important difference compared to ampicillin/clavulanate. Kinetic analyses of SHV-1 and three SHV R244 (-S, -Q, and -L) variants revealed the Ki for sulbactam was significantly elevated for the R244 variants, but the partition ratios, kcat/kinact, were markedly reduced (13 000 --> beta-lactamase was unmodified at 15 min. A parallel experiment with R244S demonstrated 70 and 88 +/- 3 Da fragments of sulbactam covalently attached to the beta-lactamase. We also observed that the Ki values of penems 1 and 2 were increased for R244 variants compared to those for SHV; however, these inhibitors effectively restored ampicillin susceptibility in vitro. Compared to that of sulbactam, the kcat/kinact values of penems for SHV-1 and R244S were low (beta-lactamase inactivators differently, but resistance can be overcome by designing penem inhibitors with strategic chemical properties that improve affinity and impair turnover.

  5. Comparison of detection methods for extended-spectrum beta-lactamases in Escherichia coli strains

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    Ewelina Kałużna

    2014-06-01

    Full Text Available Introduction: Detection of extended-spectrum beta-lactamases (ESBLs could be a major challenge for microbiologists – the difficulties arise mainly from the phenotypic differences among strains.Materials and Methods: Evaluation of ESBLs was performed on 42 strains of E. coli by: 1 DDST on MHA, 2 DDST on MHA with cloxacillin, 3 CT on MHA, according to CLSI, 4 CT on MHA with cloxacillin, 5 Etest ESBL (AB Biodisk, 6 CHROMagarTM ESBL (GRASO, 7 ChromID® ESBL (bioMérieux, and 8 automatic system VITEK2 ESBL test (bioMérieux.Result: Positive results were obtained for 20 strains using method 1, for 18 strains using method 2, 17 by method 3, 14 by method 4, 11 by method 5, 39 by method 6, 40 by method 7, and 15 by method 8. Using Etest ESBL 6.0 non-determinable results were obtained. The most consistent results were obtained when comparing the results of method 3 with results of method 2 (97.6%, and comparing the results obtained using methods 3 and 8 (95.2%.Conclusions: Based on our study we conclude that the chromogenic media can only be used as a screening method for the detection of ESBLs in E. coli rods. Etest is less useful compared to other phenotype methods, due to the impossibility of obtaining results for all the tested strains. Adding cloxacillin to MHA does not increase the frequency of detection of ESBLs in E. coli strains. DDST seems to be the most reliable among phenotypic methods for the detection of ESBLs in E. coli rods.

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

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    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.

  7. Prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated from blood cultures in Africa.

    Science.gov (United States)

    Sangare, S A; Maiga, A I; Guindo, I; Maiga, A; Camara, N; Savadogo, S; Diallo, S; Bougoudogo, F; Armand-Lefevre, L; Andremont, A; Maiga, I I

    2015-09-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae have been isolated from many regions of the world. Epidemiological studies are being conducted in Europe, North America, and Asia. No study has however been conducted in Africa to determine the prevalence and distribution of ESBLs on the continent. This literature review aimed at describing the prevalence of ESBL-producing Enterobacteriaceae isolated from blood cultures, as well as the ESBL genes involved at the international level. Our focus was mainly on Africa. We conducted a literature review on PubMed. Articles related to our study field and published between 1996 and 2014 were reviewed and entirely read for most of them, while we only focused on the abstracts of some other articles. Relevant articles to our study were then carefully reviewed and included in the review. The prevalence of ESBL-producing Enterobacteriaceae differs from one country to another. The results of our literature review however indicate that class A ESBLs prevail over the other types. We took into consideration articles focusing on various types of samples to assess the prevalence of ESBL-producing Enterobacteriaceae, but information on isolates from blood cultures is limited. The worldwide prevalence of ESBL-producing Enterobacteriaceae has increased over time. Evidence of ESBL-producing Enterobacteriaceae can be found in all regions of the world. Studies conducted in Africa mainly focused on the Northern and Eastern parts of the continent, while only rare studies were carried out in the rest of the continent. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  8. Extended-spectrum beta-lactamases in urinary tract infections caused by Enterobacteria: understanding and guidelines for action.

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    García-Tello, A; Gimbernat, H; Redondo, C; Arana, D M; Cacho, J; Angulo, J C

    2014-12-01

    Beta-lactamases are bacterial enzymes that protect microorganisms from the lethal effects of β-lactam antibiotics. The production of beta-lactamases is the most important mechanism of resistance to these antibiotics, especially in Gram-negative bacteria. Review the magnitude of the problem of extended-spectrum beta-lactamases (ESBL) in the urological setting and present the fundamental action guidelines on the issue, the main risk factors and the prevention strategies. A structured search strategy for patient, problem, intervention, comparison and result was conducted in the PubMed-Medline database to identify the most relevant studies related to the management of patients with urinary tract infection by ESBL-producing microorganisms. We also present a caseload analysis of our center on this issue. ESBL are found in Enterobacteria, mainly Klebsiella sp. and Escherichia coli and are characterized by their hydrolytic ability compared with beta-lactam antibiotics, which entails resistance to penicillin, cephalosporin and aztreonam. They are also associated with resistance to other antibiotics. There is a high risk of infection and colonization by ESBL producers in patients with prolonged hospital stays or who required invasive devices. The prior use of antibiotics and stays in residential care are also risk factors. Prevention programs should focus on preventing nosocomial infection. It is essential that a restrictive policy on the use of antibiotics be implemented. The therapy of choice for severe infections is focused on carbapenems, although their indiscriminate use should be avoided. In uncomplicated lower urinary tract infections, fosfomycin and nitrofurantoin are the best treatment alternatives. ESBL-producing strains constitute a true global health problem. Prevention strategies should focus on nosocomial infection. We should not forget, however, that the appearance of these pathogens in community-acquired infections is increasingly frequent. Therapeutic

  9. SME-type carbapenem-hydrolyzing class A beta-lactamases from geographically diverse Serratia marcescens strains.

    Science.gov (United States)

    Queenan, A M; Torres-Viera, C; Gold, H S; Carmeli, Y; Eliopoulos, G M; Moellering, R C; Quinn, J P; Hindler, J; Medeiros, A A; Bush, K

    2000-11-01

    Three sets of carbapenem-resistant Serratia marcescens isolates have been identified in the United States: 1 isolate in Minnesota in 1985 (before approval of carbapenems for clinical use), 5 isolates in Los Angeles (University of California at Los Angeles [UCLA]) in 1992, and 19 isolates in Boston from 1994 to 1999. All isolates tested produced two beta-lactamases, an AmpC-type enzyme with pI values of 8.6 to 9.0 and one with a pI value of approximately 9.5. The enzyme with the higher pI in each strain hydrolyzed carbapenems and was not inhibited by EDTA, similar to the chromosomal class A SME-1 beta-lactamase isolated from the 1982 London strain S. marcescens S6. The genes encoding the carbapenem-hydrolyzing enzymes were cloned in Escherichia coli and sequenced. The enzyme from the Minnesota isolate had an amino acid sequence identical to that of SME-1. The isolates from Boston and UCLA produced SME-2, an enzyme with a single amino acid change relative to SME-1, a substitution from valine to glutamine at position 207. Purified SME enzymes from the U. S. isolates had beta-lactam hydrolysis profiles similar to that of the London SME-1 enzyme. Pulsed-field gel electrophoresis analysis revealed that the isolates showed some similarity but differed by at least three genetic events. In conclusion, a family of rare class A carbapenem-hydrolyzing beta-lactamases first described in London has now been identified in S. marcescens isolates across the United States.

  10. 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

    , and 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......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...

  11. Involvement of AmpG in mediating a dynamic relationship between serine beta-lactamase induction and biofilm-forming ability of Escherichia coli.

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    Mallik, Dhriti; Pal, Shilpa; Ghosh, Anindya S

    2018-04-01

    AmpG permease is implicated both in beta-lactamase induction and peptidoglycan recycling in enterobacterial isolates. Here, physiological studies using molecular genetics show that deletion of AmpG permease dramatically increases beta-lactam susceptibility even in the presence of AmpC, TEM-1 and OXA beta-lactamases. Also, there is an appreciable decrease in the biofilm-forming ability of strains lacking this protein. Expression of this permease in excess probably compromises the integrity of the bacterial cells, leading to cell lysis. Based on these results, we propose that AmpG permease may be used as a potential antibiotic target and its suppression could efficiently inhibit both beta-lactamase induction and biofilm formation.

  12. Novel ambler class A carbapenem-hydrolyzing beta-lactamase from a Pseudomonas fluorescens isolate from the Seine River, Paris, France.

    Science.gov (United States)

    Girlich, Delphine; Poirel, Laurent; Nordmann, Patrice

    2010-01-01

    A Pseudomonas fluorescens isolate (PF-1) resistant to carbapenems was recovered during an environmental survey performed with water from the Seine River (Paris). It expressed a novel Ambler class A carbapenemase, BIC-1, sharing 68 and 59% amino acid identities with beta-lactamases SFC-1 from Serratia fonticola and the plasmid-encoded KPC-2, respectively. beta-Lactamase BIC-1 hydrolyzed penicillins, carbapenems, and cephalosporins except ceftazidime and monobactams. The bla(BIC-1) gene was chromosomally located and was also identified in two other P. fluorescens strains isolated from the Seine River 3 months later.

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

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    Vlademir Vicente Cantarelli

    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.

  14. Biochemical and Structural Characterization of Mycobacterium tuberculosis beta-Lactamase with the Carbapenems Ertapenem and Doripenem

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    L Tremblay; F Fan; J Blanchard

    2011-12-31

    Despite the enormous success of {beta}-lactams as broad-spectrum antibacterials, they have never been widely used for the treatment of tuberculosis (TB) due to intrinsic resistance that is caused by the presence of a chromosomally encoded gene (blaC) in Mycobacterium tuberculosis. Our previous studies of TB BlaC revealed that this enzyme is an extremely broad-spectrum {beta}-lactamase hydrolyzing all {beta}-lactam classes. Carbapenems are slow substrates that acylate the enzyme but are only slowly deacylated and can therefore act also as potent inhibitors of BlaC. We conducted the in vitro characterization of doripenem and ertapenem with BlaC. A steady-state kinetic burst was observed with both compounds with magnitudes proportional to the concentration of BlaC used. The results provide apparent K{sub m} and k{sub cat} values of 0.18 {micro}M and 0.016 min{sup -1} for doripenem and 0.18 {micro}M and 0.017 min{sup -1} for ertapenem, respectively. FTICR mass spectrometry demonstrated that the doripenem and ertapenem acyl-enzyme complexes remain stable over a time period of 90 min. The BlaC-doripenem covalent complex obtained after a 90 min soak was determined to 2.2 {angstrom}, while the BlaC-ertapenem complex obtained after a 90 min soak was determined to 2.0 {angstrom}. The 1.3 {angstrom} diffraction data from a 10 min ertapenem-soaked crystal revealed an isomerization occurring in the BlaC-ertapenem adduct in which the original {Delta}2-pyrroline ring was tautomerized to generate the {Delta}1-pyrroline ring. The isomerization leads to the flipping of the carbapenem hydroxyethyl group to hydrogen bond to carboxyl O2 of Glu166. The hydroxyethyl flip results in both the decreased basicity of Glu166 and a significant increase in the distance between carboxyl O2 of Glu166 and the catalytic water molecule, slowing hydrolysis.

  15. Extended-spectrum beta-lactamase- and carbapenemase-producing Enterobacteriaceae among Ethiopian children

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    Legese, Melese Hailu; Weldearegay, Gebru Mulugeta; Asrat, Daniel

    2017-01-01

    Background Infections by extended-spectrum beta-lactamase- (ESBL) and carbapenem-resistant Enterobacteriaceae (CRE) are an emerging problem in children nowadays. Hence, the aim of this study was to determine the prevalence of ESBL- and carbapenemase-producing Enterobacteriaceae among children suspected of septicemia and urinary tract infections (UTIs). Methods A cross-sectional study was conducted from January to March 2014. A total of 322 study participants suspected of septicemia and UTIs were recruited. All blood and urine samples were cultured on blood and MacConkey agar. All positive cultures were characterized by colony morphology, Gram stain, and standard biochemical tests. Antimicrobial susceptibility test was performed on Muller-Hinton agar using disk diffusion. ESBL was detected using combination disk and double-disk synergy methods, and the results were compared. Carbapenemase was detected by modified Hodge method using meropenem. Data were analyzed using SPSS version 20. Results The overall prevalence of ESBL- and carbapenemase-producing Enterobacteriaceae was 78.57% (n=22/28) and 12.12%, respectively. Among the Enterobacteriaceae tested, Klebsiella pneumoniae (84.2%, n=16/19), Escherichia coli (100%, n=5/5), and Klebsiella oxytoca (100%, n=1/1) were positive for ESBL. Double-disk synergy method showed 90.9% sensitivity, 66.7% specificity, 95.2% positive predictive value, and 50% negative predictive value. Carbapenemase-producing Enterobacteriaceae were K. pneumoniae (9.09%, n=3/33) and Morganella morganii (3.03%, n=1/33). Conclusion Screening Enterobacteriaceae for ESBL production is essential for better antibiotics selection and preventing its further emergence and spread. In resource-limited settings, double-disk synergy method can be implemented for screening and confirming ESBL production. Moreover, occurrence of CRE in countries where no carbapenems are sold is worrying microbiologists as well as clinicians. Hence, identifying factors that induce

  16. Effects of beta-lactamases and omp mutation on susceptibility to beta-lactam antibiotics in Escherichia coli.

    Science.gov (United States)

    Hiraoka, M; Okamoto, R; Inoue, M; Mitsuhashi, S

    1989-01-01

    Four types of beta-lactamases consisting of a penicillinase type I (TEM-1), a penicillinase type II (OXA-1), a cephalosporinase of Citrobacter freundii, and a cephalosporinase of Proteus vulgaris were introduced into Escherichia coli MC4100 and its omp mutants, MH1160 (MC4100 ompR1) and MH760 (MC4100 ompR2), by transformation. Effects of the combination of the omp mutations and these beta-lactamases on the susceptibility of E. coli strains were studied with 15 beta-lactam antibiotics including cephalosporins, cephamycins, penicillins, imipenem, and aztreonam. The ompR1 mutant, MH1160, lacks OmpF and OmpC, and it showed reduced susceptibility to 11 of the 15 beta-lactam agents. The reduction in susceptibility to cefoxitin, moxalactam, and flomoxef was much greater than reduction in susceptibility to the other agents. When the ompR1 mutant produced the cephalosporinase of C. freundii, the susceptibility of the mutant to 12 of the 15 beta-lactam antibiotics decreased. The reduction in susceptibility of MH1160 to 10 of the 12 agents affected by the enzyme was two- to fourfold greater than that observed in MC4100. Such a synergistic effect was also observed with the cephalosporinase of P. vulgaris and ompR1 mutation against six cephalosporins, moxalactam, and aztreonam. Images PMID:2658786

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

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    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.

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

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    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.

  19. 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 prob......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 problem really is. To gain insight into the magnitude and molecular epidemiology of ESBL-producing Enterobacteriaceae in East Africa a literature search was performed in PubMed on 31 July 2015 to retrieve articles with relevant information on ESBL. Methods and results: Meta-analysis was performed...... to determine overall proportion estimate of ESBL-producing Enterobacteriaceae. A total of 4076 bacterial isolates were included in the analysis. The overall pooled proportion of ESBL-producing Enterobacteriaceae among included surveys done in East African hospitals was found to be 0. 42 (95 % CI: 0...

  20. Evolving beta-lactamase epidemiology in Enterobacteriaceae from Italian nationwide surveillance, October 2013: KPC-carbapenemase spreading among outpatients.

    Science.gov (United States)

    Giani, Tommaso; Antonelli, Alberto; Caltagirone, Mariasofia; Mauri, Carola; Nicchi, Jessica; Arena, Fabio; Nucleo, Elisabetta; Bracco, Silvia; Pantosti, Annalisa; Luzzaro, Francesco; Pagani, Laura; Rossolini, Gian Maria

    2017-08-03

    Extended-spectrum beta-lactamases (ESBLs), AmpC-type beta-lactamases (ACBLs) and carbapenemases are among the most important resistance mechanisms in Enterobacteriaceae. This study investigated the presence of these resistance mechanisms in consecutive non-replicate isolates of Escherichia coli (n = 2,352), Klebsiella pneumoniae (n = 697), and Proteus mirabilis (n = 275) from an Italian nationwide cross-sectional survey carried out in October 2013. Overall, 15.3% of isolates were non-susceptible to extended-spectrum cephalosporins but susceptible to carbapenems (ESCR-carbaS), while 4.3% were also non-susceptible to carbapenems (ESCR-carbaR). ESCR-carbaS isolates were contributed by all three species, with higher proportions among isolates from inpatients (20.3%) but remarkable proportions also among those from outpatients (11.1%). Most ESCR-carbaS isolates were ESBL-positive (90.5%), and most of them were contributed by E. coli carrying bla CTX-M group 1 genes. Acquired ACBLs were less common and mostly detected in P. mirabilis. ESCR-carbaR isolates were mostly contributed by K. pneumoniae (25.1% and 7.7% among K. pneumoniae isolates from inpatients and outpatients, respectively), with bla KPC as the most common carbapenemase gene. Results showed an increasing trend for both ESBL and carbapenemase producers in comparison with previous Italian surveys, also among outpatients. This article is copyright of The Authors, 2017.

  1. 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

    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

  2. Antibiogram Studies and Extended Spectrum Beta-lactamase Activity Profile of Salmonella-like Species Isolated from Poultry Soil of the University of Uyo, Nigeria

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    Ikpeme, E. M.

    2012-01-01

    Full Text Available Aims: The contribution of beta-lactamase activity of various bacterial species to the increased antimicrobial resistance being experienced worldwide is very scanty in the literature. This study was undertaken to investigate the antibiotic resistance pattern (antibiogram of Salmonella-like bacterial species against some antibiotics, and the role beta-lactamase assumably produced by the Salmonella-like species, played in producing resistance.Methodology and Results: The antimicrobial sensitivity test and the beta-lactamase test of the Salmonella-like species were carried out using the methods of Kirby Bauer sensitivity test and the Double Disk Synergy test respectively, following isolation and identification of the organisms from poultry soil. Results revealed that Salmonella-like species were most highly resistant to Nalidixic acid (20, 66.66%, followed by Tetracycline (19, 63.33%, Cotrimoxazole, Amoxicillin and Augmentin (18, 60%, while the least was Ofloxacin (8, 26.66%. Multiple resistance of 4 or more antibiotics among the isolates from the soil outside the broilers enclosure was observed, while there was a significant difference (P <0.05 between poultry soil and control soil. This implied that the antibiotics with the highest resistance were most often applied to the birds, the droppings of which contaminated the soil. The resistant pattern of the isolates from the control soil is lower than that from the poultry soil. Extended Spectrum Beta-lactamase activity was expressed by all the isolates against Cefotazime, while the least resistance was against mostly Cefotazime.Conclusion, significance and impact of study: It is concluded that there is a widespread Beta-lactamase activity causing antibiotic resistance by many species of bacteria as well as poultry Salmonella, thus exacerbating the global problem of antibiotic resistance and a serious health related implication for antibiotic use in poultry.

  3. [Investigation of OXA type beta-lactamases and PFGE patterns in Acinetobacter baumannii strains resistant to carbapenems].

    Science.gov (United States)

    Keyik, Serafettin; Arslan, Uğur; Türk Dağı, Hatice; Seyhan, Tuba; Fındık, Duygu

    2014-10-01

    Acinetobacter baumannii is an important opportunistic and multidrug-resistant pathogen leading to nosocomial infections. Over the last 10 years, a significant and threatening increase in resistance to carbapenems, mainly due to the dissemination of class D beta-lactamases, has been reported in A.baumannii worldwide. The most common types of beta-lactamases causing carbapenem resistance in A.baumannii are the OXA-23, OXA-24, OXA-40, OXA-58 and OXA-143 type serine beta-lactamases. The aim of this study was to investigate the presence of OXA type beta-lactamases in carbapenem-resistant A.baumannii strains and the clonal relationship between the strains. A total of 105 non-duplicate carbapenem-resistant A.baumannii strains isolated from various clinical samples (68 blood, 18 bronchoalveolar lavage, 13 drainage, 3 urine, 2 cerebrospinal fluid and 1 catheter samples) in the Microbiology Laboratories of Selcuk University, Meram (2009-2012) and Selcuklu (2007-2008) Medical School Hospitals, were included in the study. The isolates were identified by conventional methods and Phoenix 100 BD (BD Diagnostic, USA) and Vitek II (bioMerieux, France) automated systems. Carbapenem susceptibility test was performed by Kirby-Bauer disk diffusion method according to the CLSI standards. bla(OXA 23-like), bla(OXA 24-like), bla(OXA 58-like) and bla(OXA 51-like) genes were amplified by multiplex PCR assay and clonal relatedness was investigated by pulsed-field gel electrophoresis (PFGE) using ApaI enzyme. The bla(OXA 51-like) gene was determined in all carbapenem-resistant A.baumannii isolates, while the bla(OXA 23-like) and bla(OXA 58-like) genes were detected in 46.6% and 53.3% of isolates, respectively. However bla(OXA 24-like) gene was not demonstrated in any isolates. bla(OXA 23-like) gene was determined in both Meram and Selcuklu Medical School hospitals, but bla(OXA 58-like) gene was detected only in Meram Medical School hospital. PFGE analysis of the isolates revealed 32 different

  4. Role of ser-237 in the substrate specificity of the carbapenem-hydrolyzing class A beta-lactamase Sme-1.

    Science.gov (United States)

    Sougakoff, W; Naas, T; Nordmann, P; Collatz, E; Jarlier, V

    1999-08-17

    The role of the serine residue found at position 237 in the carbapenemase Sme-1 has been investigated by constructing a mutant in which Ser-237 was replaced by an alanine. The S237A mutant showed a catalytic behavior against penicillins and aztreonam very similar to that of Sme-1. By contrast, S237A was characterized by a reduced catalytic efficiency against cephems, such as cephalothin and cephaloridine. In addition, the weak activity of Sme-1 against the cephamycin cefoxitin was hardly detectable with the mutant enzyme. Finally, the Ser-237-->Ala mutation resulted in a marked decrease in catalytic activity against imipenem, showing that Ser-237 contributes to the carbapenemase activity of the class A beta-lactamase Sme-1.

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

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    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.

  6. Correlation of phenotypic tests with the presence of the blaZ gene for detection of beta-lactamase.

    Science.gov (United States)

    Ferreira, Adriano Martison; Martins, Katheryne Benini; Silva, Vanessa Rocha da; Mondelli, Alessandro Lia; Cunha, Maria de Lourdes Ribeiro de Souza da

    Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤28mm (penicillin resistant) and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥29mm), 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%), followed by MIC determination (85.5%), but the specificity of the former was low (40.0%). The nitrocefin test was the least sensitive (28.9%). However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%). The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  7. Correlation of phenotypic tests with the presence of the blaZ gene for detection of beta-lactamase

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    Adriano Martison Ferreira

    Full Text Available Abstract Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤28 mm (penicillin resistant and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥29 mm, 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%, followed by MIC determination (85.5%, but the specificity of the former was low (40.0%. The nitrocefin test was the least sensitive (28.9%. However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%. The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method.

  8. Novel Ambler class A beta-lactamase LAP-1 and its association with the plasmid-mediated quinolone resistance determinant QnrS1.

    Science.gov (United States)

    Poirel, Laurent; Cattoir, Vincent; Soares, Ana; Soussy, Claude-James; Nordmann, Patrice

    2007-02-01

    The plasmid-mediated quinolone resistance determinant QnrS1 was identified in non-clonally related Enterobacter cloacae isolates in association with a transferable narrow-spectrum beta-lactam resistance marker. Cloning experiments allowed the identification of a novel Ambler class A beta-lactamase, named LAP-1. It shares 62 and 61% amino acid identity with the most closely related beta-lactamases, TEM-1 and SHV-1, respectively. It has a narrow-spectrum hydrolysis of beta-lactams and is strongly inhibited by clavulanic acid and sulbactam and, to a lesser extent, by tazobactam. Association of the blaLAP-1 gene with the qnrS1 gene was identified in E. cloacae isolates from France and Vietnam. These genes were plasmid located and associated with similar insertion sequences but were not associated with sul1-type class 1 integrons, as opposed to the qnrA genes.

  9. Restriction of cephalosporins and control of extended spectrum beta-lactamase producing gram negative bacteria in a neonatal intensive care unit.

    Science.gov (United States)

    Murki, Srinivas; Jonnala, Sravanthi; Mohammed, Faheemuddin; Reddy, Anupama

    2010-09-01

    This interventional study with historical controls was conducted to study the effect of cephalosporin restriction on the incidence of extended spectrum beta-lactamase (ESBL) gram negative infections in neonates admitted to intensive care unit. All gram negative isolates from the blood were evaluated for beta lactamase production. The incidence of ESBL production was compared before (year 2007) and after cephalosporin restriction (year 2008). Thirty two neonates (3% of NICU admissions) in the year 2007 and fifty six (5.2%) in the year 2008, had gram negative septicemia. The incidence of ESBL gram negatives decreased by 22% (47% to 25%, P=0.03). Restriction of all class of cephalosporins significantly decreased the incidence of ESBL gram negative infections.

  10. Emergence of colistin resistance in extended-spectrum beta lactamase producing Enterobacteriaceae isolated from food animals and its public health implication: A review

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    Asinamai Athliamai Bitrus

    2018-03-01

    Full Text Available Antimicrobial resistance as a result of emergence of extended-spectrum beta lactamase producing Enterobacteriaceae is a major health problem of human and animal that requires an intensive global attention. The production of beta lactamase enzymes remains as one of the major factors contributing to the development of resistance to beta lactams. These enzymes hydrolyze the beta lactam ring of the antibiotic and render it ineffective. Extended-spectrum beta lactamase producing bacteria have the ability to develop resistance to a number of antibiotics including the carbapenem and other third generation cephalosporins. In addition, the recent emergence and dissemination of the colistin resistance determinants mcr-1, mcr-2 and mcr-3 poses a serious threat to colistin as a drug of last resort in human medicine. In this review, we utilized words such as “colistin resistance and Escherichia coli”, “Klebsiella and colistin resistance”, “colistin resistance and Salmonella” as well as “detection of mcr-1 genes in Salmonella and E. coli”. The extended-spectrum beta lactamase producing bacteria under Enterobacteriaceae that are resistant to colistin possess the ability to be transferred resistant determinants to other susceptible cells at a higher frequency. In this paper, the role of manure from food animals and how air travel contributes to the dissemination of mcr-1 haboring bacteria, resistance determinants and other metabolites that constitute a public health problem was also reviewed. It is concluded that these pathogens have significant consequences to the control of infection and plays key roles in treatment failure with colistin. [J Adv Vet Anim Res 2018; 5(1.000: 1-11

  11. A trial with IgY chicken antibodies to eradicate faecal carriage of Klebsiella pneumoniae and Escherichia coli producing extended-spectrum beta-lactamases

    OpenAIRE

    Jonsson, Anna-Karin; Larsson, Anders; Tängdén, Thomas; Melhus, Åsa; Lannergård, Anders

    2015-01-01

    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...

  12. The distribution of beta lactamase genes in Escherichia coli phylotypes isolated from diarrhea and UTI cases in northwest Iran.

    Science.gov (United States)

    Hemati, Zahra; Ghanbarpour, Reza; Alizade, Hesam

    2014-01-01

    Pathogenic Escherichia coli strains are a common cause of intestinal and extra-intestinal infections, especially in developing countries. Extended spectrum beta-lactamases (ESBLS), a heterogeneous group of plasmid-encoded beta-lactamases, are common throughout the world. The aim of the present study was to determine the phenotypic and genotypic characteristics of ESBLS produced by E. coli isolates taken from patients with diarrhea and urinary tract infections (UTI) in northwest Iran. A total of 132 E. coli isolates (92 isolates from UTI and 40 isolates from diarrheic cases) were recovered and confirmed by biochemical tests. The isolates were examined for blaTEM and blaSHV genes and phylogenetic background by two multiplex PCR assays. The isolates were tested for antibiotic susceptibility against nine antibiotic agents by the disk diffusion method. The phylogenetic analysis showed that the UTI isolates mostly fell into phylo-group B2, followed by D, while the diarrheic isolates belonged to phylo-groups D and A. Out of 92 UTI isolates, 29.3% and 17.4% possessed blaTEM and blaSHV genes, respectively. Ten diarrheic isolates were positive for blaTEM, two isolates possessed the blaSHV gene, and one isolate was positive for both genes. The UTI isolates that were positive for blaTEM and blaSHV genes mostly belonged to phylo-groups D and B2, whereas the diarrhea isolates were in phylo-groups D and A. Phylogenetic group D isolates have an accumulation of ESBLS genes in the diarrheic and UTI isolates. In both the UTI and diarrhea isolates, the maximum rate of resistance was against cefazolin, and the minimum rate of resistance was against nitrofurantoin. Twenty-four antibiotic resistance patterns were observed among the isolates. The amikacin, ciprofloxacin, cefotaxime, cefuroxime, cefazolin, gentamicin, nalidixic acid and trimethoprim/sulfamethoxazole resistance pattern was the most prevalent in the isolates that belonged to phylo-group D. The correct choice of effective

  13. Development of SYN-004, an oral beta-lactamase treatment to protect the gut microbiome from antibiotic-mediated damage and prevent Clostridium difficile infection.

    Science.gov (United States)

    Kaleko, Michael; Bristol, J Andrew; Hubert, Steven; Parsley, Todd; Widmer, Giovanni; Tzipori, Saul; Subramanian, Poorani; Hasan, Nur; Koski, Perrti; Kokai-Kun, John; Sliman, Joseph; Jones, Annie; Connelly, Sheila

    2016-10-01

    The gut microbiome, composed of the microflora that inhabit the gastrointestinal tract and their genomes, make up a complex ecosystem that can be disrupted by antibiotic use. The ensuing dysbiosis is conducive to the emergence of opportunistic pathogens such as Clostridium difficile. A novel approach to protect the microbiome from antibiotic-mediated dysbiosis is the use of beta-lactamase enzymes to degrade residual antibiotics in the gastrointestinal tract before the microflora are harmed. Here we present the preclinical development and early clinical studies of the beta-lactamase enzymes, P3A, currently referred to as SYN-004, and its precursor, P1A. Both P1A and SYN-004 were designed as orally-delivered, non-systemically available therapeutics for use with intravenous beta-lactam antibiotics. SYN-004 was engineered from P1A, a beta-lactamase isolated from Bacillus licheniformis, to broaden its antibiotic degradation profile. SYN-004 efficiently hydrolyses penicillins and cephalosporins, the most widely used IV beta-lactam antibiotics. In animal studies, SYN-004 degraded ceftriaxone in the GI tract of dogs and protected the microbiome of pigs from ceftriaxone-induced changes. Phase I clinical studies demonstrated SYN-004 safety and tolerability. Phase 2 studies are in progress to assess the utility of SYN-004 for the prevention of antibiotic-associated diarrhea and Clostridium difficile disease. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  14. Differential Binding of Co(II) and Zn(II) to Metallo-beta-Lactamase Bla2 from Bacillus anthracis

    Energy Technology Data Exchange (ETDEWEB)

    Hawk, M.; Breece, R; Hajdin, C; Bender, K; Hu, Z; Costello, A; Bennett, B; Tierney, D; Crowder, M

    2009-01-01

    In an effort to probe the structure, mechanism, and biochemical properties of metallo-{beta}-lactamase Bla2 from Bacillus anthracis, the enzyme was overexpressed, purified, and characterized. Metal analyses demonstrated that recombinant Bla2 tightly binds 1 equiv of Zn(II). Steady-state kinetic studies showed that mono-Zn(II) Bla2 (1Zn-Bla2) is active, while di-Zn(II) Bla2 (ZnZn-Bla2) was unstable. Catalytically, 1Zn-Bla2 behaves like the related enzymes CcrA and L1. In contrast, di-Co(II) Bla2 (CoCo-Bla2) is substantially more active than the mono-Co(II) analogue. Rapid kinetics and UV-vis, 1H NMR, EPR, and EXAFS spectroscopic studies show that Co(II) binding to Bla2 is distributed, while EXAFS shows that Zn(II) binding is sequential. To our knowledge, this is the first documented example of a Zn enzyme that binds Co(II) and Zn(II) via distinct mechanisms, underscoring the need to demonstrate transferability when extrapolating results on Co(II)-substituted proteins to the native Zn(II)-containing forms.

  15. Extended-spectrum beta-lactamase-producing Escherichia coli in common vampire bats Desmodus rotundus and livestock in Peru.

    Science.gov (United States)

    Benavides, J A; Shiva, C; Virhuez, M; Tello, C; Appelgren, A; Vendrell, J; Solassol, J; Godreuil, S; Streicker, D G

    2018-06-01

    Antibiotic resistance mediated by bacterial production of extended-spectrum beta-lactamase (ESBL) is a global threat to public health. ESBL resistance is most commonly hospital-acquired; however, infections acquired outside of hospital settings have raised concerns over the role of livestock and wildlife in the zoonotic spread of ESBL-producing bacteria. Only limited data are available on the circulation of ESBL-producing bacteria in animals. Here, we report ESBL-producing Escherichia coli in wild common vampire bats Desmodus rotundus and livestock near Lima, Peru. Molecular analyses revealed that most of this resistance resulted from the expression of bla CTX-M-15 genes carried by plasmids, which are disseminating worldwide in hospital settings and have also been observed in healthy children of Peru. Multilocus sequence typing showed a diverse pool of E. coli strains carrying this resistance that were not always host species-specific, suggesting sharing of strains between species or infection from a common source. This study shows widespread ESBL resistance in wild and domestic animals, supporting animal communities as a potential source of resistance. Future work is needed to elucidate the role of bats in the dissemination of antibiotic-resistant strains of public health importance and to understand the origin of the observed resistance. © 2018 Blackwell Verlag GmbH.

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

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    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.

  17. Occurrence of Multidrug Resistant Extended Spectrum Beta-Lactamase-Producing Bacteria on Iceberg Lettuce Retailed for Human Consumption

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    Natasha Bhutani

    2015-01-01

    Full Text Available Antibiotic resistance in bacteria is a global problem exacerbated by the dissemination of resistant bacteria via uncooked food, such as green leafy vegetables. New strains of bacteria are emerging on a daily basis with novel expanded antibiotic resistance profiles. In this pilot study, we examined the occurrence of antibiotic resistant bacteria against five classes of antibiotics on iceberg lettuce retailed in local convenience stores in Rochester, Michigan. In this study, 138 morphologically distinct bacterial colonies from 9 iceberg lettuce samples were randomly picked and tested for antibiotic resistance. Among these isolates, the vast majority (86% demonstrated resistance to cefotaxime, and among the resistant bacteria, the majority showed multiple drug resistance, particularly against cefotaxime, chloramphenicol, and tetracycline. Three bacterial isolates (2.17% out of 138 were extended spectrum beta-lactamase (ESBL producers. Two ESBL producers (T1 and T5 were identified as Klebsiella pneumoniae, an opportunistic pathogen with transferable sulfhydryl variable- (SHV- and TEM-type ESBLs, respectively. The DNA sequence analysis of the blaSHV detected in K. pneumoniae isolate T1 revealed 99% relatedness to blaSHV genes found in clinical isolates. This implies that iceberg lettuce is a potential reservoir of newly emerging and evolving antibiotic resistant bacteria and its consumption poses serious threat to human health.

  18. Detection of Extended Spectrum Beta-Lactamases Among Gram Negative Bacilli Recovered from Cattle Feces In Benin City, Nigeria

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    Helen Oroboghae OGEFERE

    2017-06-01

    Full Text Available This study was carried out to determine the prevalence of extended spectrum beta-lactamase (ESBL among Gram negative bacteria isolated from cattle feces in Benin City, Nigeria. A total of 250 Gram negative bacteria isolates were recovered from cattle feces and were processed microbiologically using standard techniques. Emergent colonies were identified and antibacterial susceptibility tests were determined using Kirby-Bauer disk diffusion method. All bacterial isolates were screened for the presence of ESBL using the double-disc synergy method. A total of 37 (14.8% isolates were positive for ESBL, with 33 (13.2% indicated by ceftazidime, while only 4 (1.6% were indicated by both ceftazidime and cefotaxime (P < 0.0001. Of the Gram negative bacterial isolates recovered, Salmonella species was the most prevalent ESBL-producer with 55.0% prevalence (P = 0.0092, while no isolate of Pseudomonas aeruginosa produced ESBL. ESBL-positive isolates showed poor susceptibility to the tested antibacterial agents in comparison with non-ESBL-producers and imipenem was the most active antibiotic. The prevalence of ESBL among Gram negative bacilli recovered from cattle feces was 14.8%. The study advises prudent use of antibiotics in the treatment of cattle and harps on improved hygiene in managing cattle, as they are potential reservoirs of ESBL-producing organisms.

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

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    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.

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

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    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.

  1. Nosocomial extended-spectrum beta-lactamase-producing Klebsiella pneumoniae bacteremia in hemodialysis patients and the implications for antibiotic therapy.

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    Yang, Chih-Chao; Wu, Chien-Hsing; Lee, Chien-Te; Liu, Han-Tsung; Chen, Jin-Bor; Chiu, Chien-Hua; Chen, Chih-Hung; Chuang, Feng-Rong

    2014-11-01

    In the face of increasing treatment options for extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-Kp) hemodialysis (HD) access-related bacteremia, the difference in clinical effectiveness between ertapenem and flomoxef remains unclear. We conducted this retrospective study to determine their efficacies and treatment outcomes. Patients on maintenance HD with fistula-, graft-, or catheter-related ESBL-Kp bacteremia were enrolled. Data related to clinical features and antibiotic treatments were collected. Outcome was determined by mortality resulting from bacteremia during the 14-day period after the collection of the first positive blood culture for flomoxef-susceptible ESBL-Kp. The 64 patients studied had severe septicemia as determined by the Pitt bacteremia score; 50% (32/64) were in the intensive care unit (ICU) at the time of bacteremia. Old age (>65 years; 57.8%), malnutrition (albumin30 days; 75%) were also highly prevalent. The study population comprised nine fistula-, 10 graft-, and 45 HD catheter-related bacteremia cases, and the mortality rate was high (38/64, 59.4%). The mortality rate was significantly higher in the flomoxef treatment group than in the ertapenem treatment group (22/30, 73% vs. 16/34, 47%, pflomoxef use (odds ratio (OR) 2.52, 95% confidence interval (CI) 1.34-35.17) and Pitt bacteremia score (OR 4.37, 95% CI 1.28-5.26) were independently associated with mortality. In accordance with our previous study, our results have demonstrated the inferiority of flomoxef to carbapenems in the treatment of HD access-related ESBL-Kp bacteremia and provide an insight into the possibility of using ertapenem rather than flomoxef as an initial or de-escalating therapy for infections caused by ESBL-producing bacteria.

  2. Isolation and identification of extended-spectrum beta-lactamase (ESBL-producing Escherichia coli from brolier in Erbil, Iraq

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    M.N. Al-Sharook

    2017-06-01

    Full Text Available Extended-spectrum beta-lactamase-producing Escherichia coli isolated from slaughtered broilers in retail market that sell live chickens in Erbil city, Iraq. Forty-one cloacal fecal samples from broiler caecum were investigated from January to April 2016. ESBLs strains were isolated using MacConkey agar supplemented with cefotaxime 1 mg/l and the isolates were identified phynotypically by biochemical tests, TBX agar and VITEK-2 compact system. A total of 34 Escherichia coli and 4 Proteus mirabilis were analysed for determination of ESBL/AmpC by disc diffusion test using antimicrobial 68DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + ESBL inhibitor, cefpodoxime + AmpC inhibitor and cefpodoxime + ESBL inhibitor + AmpC inhibitor and 67DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + clavulanate, ceftazidime, ceftazidime + clavulanate, cefotaxime and cefotaxime + clavulanate. The phenotypic results showed that in group 68DC discs 23.7% E. coli were resistant to cefpodoxime and in group 67DC discs 73.7% of E. coli and 7.9% of P. mirabilis were resistance to one or more of the cefpodoxime, ceftazidime and ceftazidime. Final results revealed that 78.0% of samples were ESBLs/ AmpC positive. This study is the first examination to determine phenorypically E. coli producing ESBLs/AmpC in broiler chickens in Iraq. Conclusion, the healthy broiler can be a major source of ESBLs/AmpC and the possibility that transmitted to humans through the food chain, direct contact and the surrounding environment raises the concerns about public health and safety of poultry meat and the negative consequences of drug therapy that causes the spread of antibiotic resistance.

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

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    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Extended-spectrum beta-lactamase (ESBL)-positive Enterobacteriaceae in municipal sewage and their emission to the environment.

    Science.gov (United States)

    Korzeniewska, Ewa; Harnisz, Monika

    2013-10-15

    The spread of Gram-negative bacteria with plasmid-borne extended-spectrum beta-lactamases (ESBLs) has become a worldwide problem. Their prevalence is increasing, both in hospitals and in the environment. The aim of this study was to investigate the presence of ESBL-positive Enterobacteriaceae in municipal sewage and their emission to the ambient air and the river receiving effluent from wastewater treatment plant (WWTP). In the group of 455 isolated strains, up to 19.8% (90 isolates) were phenotypic ESBL-producers. They were detected in the 63 (100%) of sewage samples analyzed, 7 (33.3%) of river water and in 10 (23.8%) of air samples collected at the WWTP area. The plasmid-mediated genes encoding beta-lactams resistance were detected in almost 10% out of bacteria of the WWTP's final effluents and in above 32% out of bacteria of air at the WWTP area. It confirms that those genes are released into the environment, which might facilitate further dissemination among environmental bacteria. Moreover, genes encoding antibiotic resistance were shown to be transferrable to an Escherichia coli recipient strain, which indicates a high possibility of horizontal gene transfer among strains of different genera within the sewage and environmental samples. This study demonstrated that despite the treatment, the municipal sewage may be a reservoir of antibiotic-resistant microorganisms and plasmid-mediated antibiotic resistance genes. This may pose a public health risk, which requires future evaluation and control. Copyright © 2013 Elsevier Ltd. All rights reserved.

  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. Case of Meningitis in a Neonate Caused by an Extended-Spectrum-Beta-Lactamase-Producing Strain of Hypervirulent Klebsiella pneumoniae

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    Khalit S. Khaertynov

    2017-08-01

    Full Text Available Klebsiella pneumoniae is one of the most important infectious agents among neonates. This pathogen has a potential to develop an increased antimicrobial resistance and virulence. The classic non-virulent strain of K. pneumoniae, producing an extended-spectrum beta-lactamases (ESBL, is associated with nosocomial infection mainly in preterm neonates. Hypervirulent K. pneumoniae strains are associated with invasive infection among previously healthy ambulatory patients, and most of them exhibit antimicrobial susceptibility. During the last few years, several cases of diseases caused by hypervirulent K. pneumoniae producing ESBL have been registered in different geographical regions of the world. However, reports of such cases in neonates are rare. Here, we reported that this pathogen can cause pyogenic meningitis in full-term neonate with poor prognosis. A previously healthy, full-term, 12-day-old neonate was admitted to the infectious diseases hospital with suspected meningitis. The clinical symptoms included loss of appetite, irritability, fever, seizures, and a bulging anterior fontanelle. The analysis of the cerebrospinal fluid confirmed the diagnosis of meningitis. Blood and cerebrospinal fluid cultures were positive for K. pneumoniae, producing ESBL. K. pneumoniae isolates were resistant to aminopenicillins, 3rd generation cephalosporins but were sensitive to imipenem and meropenem. The “string test” was positive. The study of the virulence factors of K. pneumoniae by PCR revealed the presence of the rmpA gene. A combination of K. pneumoniae virulence and drug resistance complicated by cerebral oedema led to the death of the neonate. We concluded that both the risk of developing severe forms of infection and the outcome of the disease due to K. pneumonia are associated with the phenotypic features of the pathogen such as its antibiotic susceptibility and virulence factors. Emergence of the ESBL-producing strain of hypervirulent K

  7. Ventilator-associated pneumonia caused by carbapenem-resistant Enterobacteriaceae carrying multiple metallo-beta-lactamase genes

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    Dwivedi Mayank

    2009-07-01

    Full Text Available Context: Ventilator-associated pneumonia (VAP is a leading nosocomial infection in the intensive care unit (ICU. Members of Enterobacteriaceae are the most common causative agents and carbapenems are the most commonly used antibiotics. Metallo-beta-lactamase (MBL production leading to treatment failure may go unnoticed by routine disc diffusion susceptibility testing. Moreover, there is not much information on association of MBL-producing Enterobacteriaceae with ICU-acquired VAP. Therefore, a study was undertaken to find out the association of MBL-producing Enterobacteriaceae with VAP. Settings: This study was conducted in a large tertiary care hospital of North India with an eight-bed critical care unit. Materials and Methods: The respiratory samples (bronchoalveolar lavage, protected brush catheter specimens and endotracheal or transtracheal aspirates obtained from VAP patients (during January 2005-December 2006 were processed, isolated bacteria identified and their antibiotic susceptibilities tested as per standard protocols. The isolates of Enterobacteriaceae resistant to carbapenem were subjected to phenotypic and genotypic tests for the detection of MBLs. Results: Twelve of 64 isolates of Enterobacteriaceae were detected as MBL producers, bla IMP being the most prevalent gene. Additionally, in three strains, simultaneous coexistence of multiple MBL genes was detected. Conclusion: The coexistence of multiple MBL genes in Enterobacteriaceae is an alarming situation. As MBL genes are associated with integrons that can be embedded in transposons, which in turn can be accommodated on plasmids thereby resulting in a highly mobile genetic apparatus, the further spread of these genes in different pathogens is likely to occur.

  8. Molecular characterization of extended-spectrum beta-lactamases (ESBLs) produced by clinical isolates of Acinetobacter baumannii in Saudi Arabia.

    Science.gov (United States)

    Alyamani, Essam J; Khiyami, Mohamed A; Booq, Rayan Y; Alnafjan, Basel M; Altammami, Musaad A; Bahwerth, Fayez S

    2015-08-20

    Acinetobacter baumannii is a common opportunistic pathogen that causes major nosocomial infections in hospitals. In this study, we hypothesized a high prevalence of A. baumanni ESBL (extended-spectrum beta-lactamase) among all collected isolates. A. baumannii isolates (n = 107) from ICU (Intensive care unit) of local hospitals in Makkah were phenotypically and genotypically characterized. The identity and antibiotic susceptibility of A. baumannii strains were determined using the Vitek-2 system. The identified ESBL producers were further analyzed by PCR and sequencing followed by MLST typing. bla TEM , bla SHV , and the bla CTX-M-group genes 1, 2, 8, 9, and 25 were investigated. Furthermore, bla OXA51-like and bla OXA23-like genes were also examined in the carbapenem-resistant A. baumannii isolates. Our data indicated a high prevalence of A. baumannii ESBL producers among the collected strains. Of the 107 A. baumannii isolates, 94 % were found to be resistant to cefepime and ceftazidime, and aztreonam using the Vitek 2 system. The genes detected encoded TEM, OXA-51-like and OXA-23-like enzymes, and CTX-M-group proteins 1, 2, 8, 9, and 25. MLST typing identified eight sequence type (ST) groups. The most dominant STs were ST195 and ST557 and all of them belong to worldwide clonal complex (CC) 2. This study has shown that there is a high prevalence of antimicrobial resistance in A. baumannii. The diversity of STs may suggest that new ESBL strains are constantly emerging. The molecular diversity of the ESBL genes in A. baumannii may have contributed to the increased antimicrobial resistance among all isolates.

  9. Successful elimination of extended-spectrum beta-lactamase (ESBL)-producing nosocomial bacteria at a neonatal intensive care unit.

    Science.gov (United States)

    Szél, Borbála; Reiger, Zsolt; Urbán, Edit; Lázár, Andrea; Mader, Krisztina; Damjanova, Ivelina; Nagy, Kamilla; Tálosi, Gyula

    2017-06-01

    Extended-spectrum beta-lactamase (ESBL)-producing Gram-negative bacteria are highly dangerous to neonates. At our Neonatal Intensive Care Unit (NICU), the presence of these bacteria became so threatening in 2011 that immediate intervention was required. This study was conducted during a nearly two-year period consisting of three phases: retrospective (9 months), educational (3 months) and prospective (9 months). Based on retrospective data analysis, a complex management plan was devised involving the introduction of the INSURE protocol, changes to the antibiotic regimen, microbiological screening at short intervals, progressive feeding, a safer bathing protocol, staff hand hygiene training and continuous monitoring of the number of newly infected and newly colonized patients. During these intervals, a total of 355 patients were monitored. Both ESBL-producing Enterobacter cloaceae and Klebsiella pneumoniae were found (in both patients and environmental samples). In the prospective period a significant reduction could be seen in the average number of both colonized (26/167 patients; P=0.029) and infected (3/167 patients; P=0.033) patients compared to data from the retrospective period regarding colonized (72/188 patients) and infected (9/188 patients) patients. There was a decrease in the average number of patient-days (from 343.72 to 292.44 days per months), though this difference is not significant (P=0.058). During the prospective period, indirect hand hygiene compliance showed a significant increase (from the previous 26.02 to 33.6 hand hygiene procedures per patient per hospital day, Pinfections were rolled back successfully in a multi-step effort that required an interdisciplinary approach.

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

    Science.gov (United States)

    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.

  11. Prevalence and characterization of extended-spectrum beta-lactamases in Klebsiella pneumoniae in Algiers hospitals (Algeria).

    Science.gov (United States)

    Messai, Y; Iabadene, H; Benhassine, T; Alouache, S; Tazir, M; Gautier, V; Arlet, G; Bakour, R

    2008-07-01

    To determine the prevalence and the diversity of extended-spectrum beta-lactamases (ESBLs) in 196 Klebsiella pneumoniae clinical isolates collected from three hospitals in Algiers. Antibiograms were done on Mueller-Hinton agar plates with the disc-diffusion method and MICs were determined by agar-dilution method. Mating experiments were performed in agar medium. Plasmid DNA was extracted by the alcalin-lysis method. Total DNA was extracted with a Qiagen mini kit and screened for bla(TEM) and bla(CTX-M) genes by PCR. Linkage of bla(CTX-M) genes with insertion sequence ISEcp1B and class 1 integrons was investigated by PCR. PCR products were sequenced by the Sanger method. The epidemiological relationships between ESBL-producing K. pneumoniae isolates were analyzed by ERIC-PCR. Thirty-nine (19.9%) isolates were found to produce ESBLs belonging to CTX-M-1 group and TEM penicillinases (CTX-M-3, CTX-M-15 and TEM-1). ERIC-PCR analysis showed that the isolates are genetically unrelated. The bla(TEM) and bla(CTX-M) genes as well as aminoglycosides and sulfonamides resistance determinants were found located in self-transferable plasmids of approximately 85 kb. The class 1 integrons and the insertion sequence ISEcp1B were present in the isolates and in their transconjugants. ISEcp1B was found genetically linked to the bla(CTX-M) genes and located 127bp upstream, with the presence of the V and W sequences. The study revealed a high rate of ESBL-producing K. pneumoniae in Algerian hospitals, resulting from horizontal dissemination of mobile bla(CTX-M) genes.

  12. 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.

  13. In vitro Effectiveness of Commercial Bacteriophage Cocktails on Diverse Extended-Spectrum Beta-Lactamase Producing Escherichia coli Strains.

    Science.gov (United States)

    Gundogdu, Aycan; Bolkvadze, Darajen; Kilic, Huseyin

    2016-01-01

    The objective of this study is to determine the in vitro susceptibility of Georgian bacteriophage cocktails on multidrug resistant (MDR) extended-spectrum beta-lactamase producing Escherichia coli (ESBL-EC) isolated from patients' blood and urine cultures. A total of 615 E. coli isolates were included in this study. Phene Plate (PhP)-typing and phylogenetic grouping were used for the typing. Antimicrobial resistance profiles and ESBL production of all isolates were confirmed according to Clinical and Laboratory Standards Institute (CLSI) criteria. The activities of four bacteriophage cocktails (Enko-phage, SES-bacteriophage, Pyo-bacteriophage, and Intesti-bacteriophage) were determined against 142 ESBL-EC using in vitro spot tests. According to this, Enko-phage were active against 87.3% of the tested strains while that ratio was 81.7% for Intesti-bacteriophage, 81.7% for Pyo-bacteriophage, and 59.2% for SES-bacteriophage cocktails. Based on the contingency tests, the phage cocktails were observed to be statistically significantly ( p < 0.001) more effective on ESBL-EC strains belonging to phylogenetic groups D and B2. The employed phage cocktails were found to be affective against all tested resistant types. These results are promising especially for the infections that are caused by MDR pathogens that are difficult to treat. As this is a preliminary step to the potential clinical trials to be designed for the country, in vitro confirmation of their success on a MDR ESBL-EC collection should be accepted as an initial action, which is encouraging to consider clinical trials of phage therapy especially in countries which are not introduce phage therapy.

  14. Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in local and imported poultry meat in Ghana.

    Science.gov (United States)

    Eibach, Daniel; Dekker, Denise; Gyau Boahen, Kennedy; Wiafe Akenten, Charity; Sarpong, Nimako; Belmar Campos, Cristina; Berneking, Laura; Aepfelbacher, Martin; Krumkamp, Ralf; Owusu-Dabo, Ellis; May, Jürgen

    2018-04-01

    Antibiotic use in animal husbandry has raised concerns on the spread of resistant bacteria. Currently animal products are traded globally with unprecedented ease, which has been challenging the control of antimicrobial resistance. This study aims to detect and characterize extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae from imported and locally produced poultry products sold in Ghana. Local and imported chicken meat was collected from 94 stores and markets throughout Kumasi (Ghana) and cultured on selective ESBL screening agar. Phenotypic ESBL-producing E. coli and K. pneumoniae isolates were confirmed by combined disc test and further characterized by antibiotic susceptibility testing, amplification of the bla CTX-M , bla TEM and bla SHV genes as well as multilocus sequence typing (MLST) and linked to the country of origin. Out of 200 meat samples, 71 (36%) samples revealed 81 ESBL-producing isolates (46 E. coli and 35 K. pneumoniae), with 44% (30/68) of local poultry and 31% (41/132) of imported products being contaminated. Most ESBL-producing isolates harboured the bla CTX-M-15 gene (61/81, 75%) and the dominant Sequence Types (ST) were ST2570 (7/35, 20%) among K. pneumoniae and ST10 (5/46, 11%) among E. coli. High numbers of ESBL-producing bacteria, particularly on local but also imported poultry meat, represent a potential source for human colonization and infection as well as spread within the community. Surveillance along the poultry production-food-consumer chain would be a valuable tool to identify sources of emerging multidrug resistant pathogens in Ghana. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Plasmid-mediated AmpC-type beta-lactamase isolated from Klebsiella pneumoniae confers resistance to broad-spectrum beta-lactams, including moxalactam.

    Science.gov (United States)

    Horii, T; Arakawa, Y; Ohta, M; Ichiyama, S; Wacharotayankun, R; Kato, N

    1993-01-01

    Klebsiella pneumoniae NU2936 was isolated from a patient and was found to produce a plasmid-encoded beta-lactamase (MOX-1) which conferred resistance to broad spectrum beta-lactams, including moxalactam, flomoxef, ceftizoxime, cefotaxime, and ceftazidime. Resistance could be transferred from K. pneumoniae NU2936 to Escherichia coli CSH2 by conjugation with a transfer frequency of 5 x 10(-7). The structural gene of MOX-1 (blaMOX-1) was cloned and expressed in E. coli HB101. The MIC of moxalactam for E. coli HB101 producing MOX-1 was > 512 micrograms/ml. The apparent molecular mass and pI of this enzyme were calculated to be 38 kDa and 8.9, respectively. Hg2+ and Cu2+ failed to block enzyme activity, and the presence of EDTA in the reaction buffer did not reduce the enzyme activity. However, clavulanate and cloxacillin, serine beta-lactamase inhibitors, inhibited the enzyme activity competitively (Kis = 5.60 and 0.35 microM, respectively). The kinetic study of MOX-1 suggested that it effectively hydrolyzed broad-spectrum beta-lactams. A hybridization study confirmed that blaMOX-1 is encoded on a large resident plasmid (pRMOX1; 180 kb) of strain NU2936. By deletion analysis, the functional region was localized within a 1.2-kb region of the plasmid. By amino acid sequencing, 18 of 33 amino acid residues at the N terminus of MOX-1 were found to be identical to those of Pseudomonas aeruginosa AmpC. These findings suggest that MOX-1 is a plasmid-mediated AmpC-type beta-lactamase that provides enteric bacteria resistance to broad-spectrum beta-lactams, including moxalactam. Images PMID:8517725

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

    International Nuclear Information System (INIS)

    Blakeley, Matthew P.; Chen, Yu; Afonine, Pavel

    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.

  17. A putative low-molecular-mass penicillin-binding protein (PBP) of Mycobacterium smegmatis exhibits prominent physiological characteristics of DD-carboxypeptidase and beta-lactamase.

    Science.gov (United States)

    Bansal, Ankita; Kar, Debasish; Murugan, Rajagopal A; Mallick, Sathi; Dutta, Mouparna; Pandey, Satya Deo; Chowdhury, Chiranjit; Ghosh, Anindya S

    2015-05-01

    DD-carboxypeptidases (DD-CPases) are low-molecular-mass (LMM) penicillin-binding proteins (PBPs) that are mainly involved in peptidoglycan remodelling, but little is known about the dd-CPases of mycobacteria. In this study, a putative DD-CPase of Mycobacterium smegmatis, MSMEG_2433 is characterized. The gene for the membrane-bound form of MSMEG_2433 was cloned and expressed in Escherichia coli in its active form, as revealed by its ability to bind to the Bocillin-FL (fluorescent penicillin). Interestingly, in vivo expression of MSMEG_2433 could restore the cell shape oddities of the septuple PBP mutant of E. coli, which was a prominent physiological characteristic of DD-CPases. Moreover, expression of MSMEG_2433 in trans elevated beta-lactam resistance in PBP deletion mutants (ΔdacAdacC) of E. coli, strengthening its physiology as a dd-CPase. To confirm the biochemical reason behind such physiological behaviours, a soluble form of MSMEG_2433 (sMSMEG_2433) was created, expressed and purified. In agreement with the observed physiological phenomena, sMSMEG_2433 exhibited DD-CPase activity against artificial and peptidoglycan-mimetic DD-CPase substrates. To our surprise, enzymic analyses of MSMEG_2433 revealed efficient deacylation for beta-lactam substrates at physiological pH, which is a unique characteristic of beta-lactamases. In addition to the MSMEG_2433 active site that favours dd-CPase activity, in silico analyses also predicted the presence of an omega-loop-like region in MSMEG_2433, which is an important determinant of its beta-lactamase activity. Based on the in vitro, in vivo and in silico studies, we conclude that MSMEG_2433 is a dual enzyme, possessing both DD-CPase and beta-lactamase activities. © 2015 The Authors.

  18. Biofilm and metallo beta-lactamase production among the strains of Pseudomonas aeruginosa and Acinetobacter spp. at a Tertiary Care Hospital in Kathmandu, Nepal

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    Bandana Baniya

    2017-11-01

    Full Text Available Abstract Introduction Pseudomonas aeruginosa and Acinetobacter spp. are found to be associated with biofilm and metallo-β-lactamase production and are the common causes of serious infections mainly in hospitalized patients. So, the main aims of this study were to determine the rates of biofilm production and metallo beta-lactamase production (MBL among the strains of Pseudomonas aeruginosa and Acinetobacter spp. isolated from hospitalized patients. Methods A total of 85 P. aeruginosa isolates and 50 Acinetobacter spp. isolates isolated from different clinical specimens from patients admitted to Shree Birendra Hospital, Kathmandu, Nepal from July 2013 to May 2014 were included in this study. The bacterial isolates were identified with the help of biochemical tests. Modified Kirby-Bauer disc diffusion technique was used for antimicrobial susceptibility testing. Combined disc diffusion technique was used for the detection of MBL production, while Congo red agar method and tube adherence method were used for detection of biofilm production. Results Around 16.4% of P. aeruginosa isolates and 22% of the strains of Acinetobacter spp. were metallo β-lactamase producers. Out of 85 P. aeruginosa isolates, 23 (27.05% were biofilm producers according to tube adherence test while, only 13 (15.29% were biofilm producers as per Congo red agar method. Similarly, out of 50 Acinetobacter spp. 7 (14% isolates were biofilm producers on the basis of tube adherence test, while only 5 (10% were positive for biofilm production by Congo red agar method. Highest rates of susceptibility of P. aeruginosa as well as Acinetobacter spp. were seen toward colistin. Conclusion In our study, biofilm production and metallo beta-lactamase production were observed among Pseudomonas aeruginosa and Acinetobacter spp. However, no statistically significant association could be established between biofilm production and metallo beta-lactamase production.

  19. Sensitivity and specificity of various beta-lactam antibiotics and phenotypical methods for detection of TEM, SHV and CTX-M extended-spectrum beta-lactamases.

    Science.gov (United States)

    Bedenic, B; Vranes, J; Mihaljevic, Lj; Tonkic, M; Sviben, M; Plecko, V; Kalenic, S

    2007-04-01

    The aim of this study was to compare the sensitivity and specificity of six different beta-lactam antibiotics using five phenotypical tests for detection of extended spectrum beta-lactamases (ESBLs) based on synergism of beta-lactam antibiotics and clavulanate. Experiments were performed on a set of 80 Klebsiella pneumoniae strains and 105 Escherichia coli strains with previously characterized ESBLs (SHV, TEM and CTX-M). ESBLs were detected by five different phenotypical methods: MIC (minimum inhibitory concentration) determination of beta-lactam antibiotics with and without clavulanate, double-disk synergy test (DDST), inhibitor-potentiated disk-diffusion test (IPDDT), CLSI-Clinical and Laboratory Standard Institution (former NCCLS) combined-disk-test, and modified MAST-disk-diffusion test (MAST-DD-test). Seven antibiotics were tested as indicators of ESBL production: ceftazidime, cefotaxime, ceftriaxone, aztreonam, ceftibuten, cefpodoxime and cefepime. Ceftazidime and aztreonam were the best indicators for SHV-5, SHV-12 and TEM beta-lactamases whereas cefotaxime and ceftriaxone were the most sensitive in detection of SHV-2 and CTX-M beta-lactamases in DDST, IPDDT and CLSI test. MIC determination of beta-lactam antibiotics with and without clavulanate was the most sensitive method. DDST was the least sensitive test. Double-disk synergy test, which is the most frequently used test for detection of ESBLs in routine laboratories, was the least sensitive independently of the indicator antibiotic. Since MIC determination is a very laborious and time consuming method, we would recommend the NCCLS combined disk test or IPDD test for detection of ESBLs in routine laboratories with 5 mm zone augmentation breakpoint.

  20. Detection of Beta-lactamase gene in the culturable bacteria isolated from agricultural, pasture and mining soils around mines in Hamedan, Iran

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    Nayereh Younessi

    2017-09-01

    Full Text Available Introduction: Growing evidence exists that agriculture affects antibiotic resistance in human pathogens. Beta-lactam antibiotics are the most commonly used antimicrobial agents in many countries. The abundance of beta-lactamase encoding genes can be used as an indicator of antibiotic resistance in the environment. So, to determine the beta-lactamase resistance genes, the abundance of culturable bacteria having bla-TEM genesin the soils under different land uses wasexamined. Materials and methods: 44 Gram-positive and 34 Gram-negative bacteria plated on nutrient agar were isolated from agricultural, pasture and mining soils and selected to study the presence of TEM-class gene using PCR amplification. Antibiotic sensitivity test of bla-TEM+isolateswas done adopting the Kirby-Bauer disk diffusion method and antibiotic discs used were: ampicillin, amoxicillin, vancomicin, streptomycin, tetracycline and gentamicin. Finally, five multi-drug resistant and bla-TEM+ isolates were identified using universal primers. Results: The highest level of beta-lactamase genes was observed in the Gram-positive and Gram-negative isolates from the pasture soils. In the agricultural and mining soils, a high abundance of bla-TEM+ isolateswasfound which also showed resistance to beta-lactam antibiotics. The identified multi-drug resistant and bla-TEM+ isolates were from these genera: Achromobacter, Bacillus, Brevibacillus, Aminobacter and Brevundimonas. Discussion and conclusion: The high number of bla-TEM+ bacteria in all the soils may be attributed to the other important feature of bla genes which is their capability to extrude toxic compounds like heavy metals in contaminated environments. Sensitivity of some bla-TEM+ bacteria to beta-lactam antibiotics was interesting. This result shows that bla-TEM genes confer resistance to beta-lactamase inhibitors in a different degree. Some of the identified isolates were pathogen. These pathogens in soils can transfer to plants

  1. TEM and CTX-M extended-spectrum beta-lactamase in Klebsiella spp and Escherichia coli isolates from inanimate surfaces of hospital environments

    OpenAIRE

    Rivera-Jacinto, Marco; Facultad de Ciencias de la Salud, Universidad Nacional de Cajamarca. Cajamarca, Perú.; Rodríguez-Ulloa, Claudia; Facultad de Ciencias de la Salud, Universidad Nacional de Cajamarca. Cajamarca, Perú.; Flores Clavo, René; Hospital Regional de Lambayeque. Lambayeque, Perú.; Serquén López, Luis; Hospital Regional de Lambayeque. Lambayeque, Perú.; Arce Gil, Zhandra; Universidad Católica Santo Toribio de Mogrovejo. Lambayeque, Perú.

    2015-01-01

    The aim of the study was to determine the genotype of 15 ESBL strains of Enterobacteriaceae resistant to beta-lactams, isolated from inanimate surfaces and phenotypically characterized as producing extended-spectrum beta-lactamase. After evaluation and screening of the bacterial strains, a PCR was conducted to amplify fragments of 1078 bp and 544 bp corresponding to type TEM and CTX-M ESBL. Eleven strains presented both fragments at the time and only three had blaCTX-M. In conclusion, the pre...

  2. 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

  3. Molecular characteristics of travel-related extended-spectrum-beta-lactamase-producing Escherichia coli isolates from the Calgary Health Region.

    Science.gov (United States)

    Pitout, Johann D D; Campbell, Lorraine; Church, Deirdre L; Gregson, Daniel B; Laupland, Kevin B

    2009-06-01

    Extended-spectrum-beta-lactamase (ESBL)-producing Escherichia coli has recently emerged as a major risk factor for community-acquired, travel-related infections in the Calgary Health Region. Molecular characterization was done on isolates associated with infections in returning travelers using isoelectric focusing, PCR, and sequencing for bla(CTX-M)s, bla(TEM)s, bla(SHV)s, bla(OXA)s, and plasmid-mediated quinolone resistance determinants. Genetic relatedness was determined with pulsed-field gel electrophoresis using XbaI and multilocus sequence typing (MLST). A total of 105 residents were identified; 6/105 (6%) presented with hospital-acquired infections, 9/105 (9%) with health care-associated community-onset infections, and 90/105 (86%) with community-acquired infections. Seventy-seven of 105 (73%) of the ESBL-producing E. coli isolates were positive for bla(CTX-M) genes; 55 (58%) produced CTX-M-15, 13 (14%) CTX-M-14, six (6%) CTX-M-24, one (1%) CTX-M-2, one (1%) CTX-M-3, and one (1%) CTX-M-27, while 10 (10%) produced TEM-52, three (3%) TEM-26, 11 (11%) SHV-2, and four (4%) produced SHV-12. Thirty-one (30%) of the ESBL-producing E. coli isolates were positive for aac(6')-Ib-cr, and one (1%) was positive for qnrS. The majority of the ESBL-producing isolates (n = 95 [90%]) were recovered from urine samples, and 83 (87%) were resistant to ciprofloxacin. The isolation of CTX-M-15 producers belonging to clone ST131 was associated with travel to the Indian subcontinent (India, Pakistan), Africa, the Middle East, and Europe, while clonally unrelated strains of CTX-M-14 and -24 were associated with travel to Asia. Our study suggested that clone ST131 coproducing CTX-M-15, OXA-1, TEM-1, and AAC(6')-Ib-cr and clonally unrelated CTX-M-14 producers have emerged as important causes of community-acquired, travel-related infections.

  4. Carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae in HIV-infected children in Zimbabwe.

    Science.gov (United States)

    Wilmore, S M S; Kranzer, K; Williams, A; Makamure, B; Nhidza, A F; Mayini, J; Bandason, T; Metcalfe, J; Nicol, M P; Balakrishnan, I; Ellington, M J; Woodford, N; Hopkins, S; McHugh, T D; Ferrand, R A

    2017-05-01

    Antimicrobial resistance is an emerging global health issue. Data on the epidemiology of multidrug-resistant organisms are scarce for Africa, especially in HIV-infected individuals who often have frequent contact with healthcare. We investigated the prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) carriage in stool among HIV-infected children attending an HIV outpatient department in Harare, Zimbabwe. We recruited children who were stable on antiretroviral therapy (ART) attending a HIV clinic from August 2014 to June 2015. Information was collected on antibiotic use and hospitalization. Stool was tested for ESBL-E through combination disc diffusion. API20E identification and antimicrobial susceptibility was performed on the positive samples followed by whole genome sequencing. Stool was collected from 175/202 (86.6 %) children. Median age was 11 [inter-quartile range (IQR) 9-12] years. Median time on ART was 4.6 years (IQR 2.4-6.4). ESBL-Es were found in 24/175 samples (13.7 %); 50 % of all ESBL-Es were resistant to amoxicillin-clavulanate, 100 % to co-trimoxazole, 45.8 % to chloramphenicol, 91.6 % to ceftriaxone, 20.8 % to gentamicin and 62.5 % to ciprofloxacin. ESBL-Es variously encoded CTX-M, OXA, TEM and SHV enzymes. The odds of ESBL-E carriage were 8.5 times (95 % CI 2.2-32.3) higher in those on ART for less than one year (versus longer) and 8.5 times (95 % CI 1.1-32.3) higher in those recently hospitalized for a chest infection. We found a 13.7 % prevalence of ESBL-E carriage in a population where ESBL-E carriage has not been described previously. Antimicrobial resistance (AMR) in Africa merits further study, particularly given the high HIV prevalence and limited diagnostic and therapeutic options available.

  5. Genetic characterization of antibiotic resistance in enteropathogenic Escherichia coli carrying extended-spectrum beta-lactamases recovered from diarrhoeic rabbits.

    Science.gov (United States)

    Poeta, P; Radhouani, H; Gonçalves, A; Figueiredo, N; Carvalho, C; Rodrigues, J; Igrejas, G

    2010-05-01

    A total of 52 Escherichia coli strains isolated from diarrhoeic rabbits were investigated for their enteropathogenic E. coli (EPEC) pathotype by PCR amplification of eae and bfp virulence genes. A total of 22 EPEC isolates were identified, serotyped and studied for antibiotic resistance and screened for the detection of extended-spectrum beta-lactamases (ESBLs). The EPEC isolates belonged to three serogroups (O26, O92 and O103). The most common serogroup (O103:K-:H2) was observed among 17 EPEC strains, the O92:K-serogroup in three isolates (the antibiotic sensitive ones) and the remaining O26:K-serogroup in two isolates (the ESBLs isolates). Resistances to ampicillin and tetracycline were the most frequent and detected followed by resistance to nalidixic acid, streptomycin, trimethoprim-sulphamethoxazole, cefoxitin, gentamicin and ciprofloxacin. All the isolates were sensitive for amikacin, ceftazidime, aztreonam, imipenem, chloramphenicol, tobramycin and amoxicillin + clavulanic acid. Two isolates recovered from two adult animals showed an intermediate susceptibility to cefotaxime, and a positive screening test for ESBL was demonstrated in both. The bla(TEM) gene was demonstrated in the majority of ampicillin-resistant isolates. The aac(3)-II or aac(3)-IV genes were detected in the four gentamicin-resistant isolates. In addition, the aadA gene was detected in 60% of streptomycin-resistant isolates. The tet(A) or tet(B) genes were identified in all tetracycline-resistant isolates. A total of nine EPEC isolates showed the phenotype SXT-resistant, and the sul1 and/or sul2 and/or sul3 genes were detected in all of them. Our findings showed that the molecular detection by the eae and bfp genes by PCR followed by serotyping is useful for monitoring trends in EPEC infections of rabbits allowing the identification of their possible reservoirs. The detection of genes involved in the resistance to antibiotics of different families in a relatively high proportion of faecal E

  6. 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

  7. Risk Factor Analysis of Ciprofloxacin-Resistant and Extended Spectrum Beta-Lactamases Pathogen-Induced Acute Bacterial Prostatitis in Korea.

    Science.gov (United States)

    Lee, Young; Lee, Dong Gi; Lee, Sang Hyub; Yoo, Koo Han

    2016-11-01

    The objectives of this study were to investigate risk factors and the incidence of ciprofloxacin resistance and extended-spectrum beta-lactamases (ESBL) in patients with acute bacterial prostatitis (ABP). We reviewed the medical records of 307 patients who were diagnosed with ABP between January 2006 and December 2015. The etiologic pathogens and risk factors for ciprofloxacin-resistant E. coli and ESBL-producing microbes, susceptibility to ciprofloxacin, and the incidence of ESBL in patients with ABP were described. History of prior urologic manipulation was an independent risk factor for ciprofloxacin-resistant (P = 0.005) and ESBL-producing microbes (P = 0.005). Advanced age (over 60 years) was an independent risk factor for ciprofloxacin-resistant microbes (P = 0.022). The ciprofloxacin susceptibility for Escherichia coli in groups without prior manipulation was documented 85.7%. For groups with prior manipulation, the susceptibility was 10.0%. Incidence of ESBL-producing microbes by pathogen was 3.8% for E. coli and 1.0% for Klebsiella pneumonia in the absence of manipulation group, and 20% and 33.3% in the presence of manipulation group, respectively. Initial treatment of ABP must consider patient's age and the possibility of prior manipulation to optimize patient treatment. With the high rate of resistance to fluoroquinolone, cephalosporins with amikacin, or carbapenems, or extended-spectrum penicillin with beta lactamase inhibitor should be considered as the preferred empirical ABP treatment in the patients with history of prior urologic manipulation.

  8. 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.

  9. Prevalence of metallo-beta-lactamase among Pseudomonas aeruginosa and Acinetobacter baumannii isolated from burn wounds and in vitro activities of antibiotic combinations against these isolates.

    Science.gov (United States)

    Altoparlak, Ulku; Aktas, Ferda; Celebi, Demet; Ozkurt, Zulal; Akcay, Mufide N

    2005-09-01

    The prevalence of metallo-beta-lactamases (MBLs) produced by isolates of Pseudomonas aeruginosa and Acinetobacter baumannii and the activities of various antmicrobial combinations against MBL producer strains were investigated. During the period from June 2003 till July 2004, 120 P. aeruginosa and 9 A. baumannii nonduplicate isolates were obtained from burn wounds. Forty strains (37 P. aeruginosa, 3 A. baumannii) were selected because of resistance to carbapenems. Screening for MBL production was performed in the latter isolates by the combined disk method which depends on comparing the zones given by disks containing imipenem with and without ethylenediaminetetraacetic acid (EDTA). Of imipenem resistant P. aeruginosa strains, 21 and 1 of A. baumannii were found metallo-beta-lactamase producers. Disk approximation studies were then performed to test for in vitro activities of various antimicrobial combinations. For a total of 21 P. aeruginosa strains, synergy was demonstrated predominantly by ciprofloxacin in combination with ceftazidime and imipenem, by ofloxacin in combination with astreonam. Against MBL producer A. baumannii strain, synergy was detected only with imipenem-ofloxacin combination. None of the combinations were antagonistic. These results suggest that MBL producing P. aeruginosa and A. baumanni strains have been introduced into burn centers, and to prevent the further spread of MBL producers, it is essential for carbapenem resistant isolates to be screened for MBLs.

  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. 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...

  12. 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, Sierd; Venema, Gerard

    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

  13. 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

    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

  14. The role of a second-shell residue in modifying substrate and inhibitor interactions in the SHV beta-lactamase: a study of ambler position Asn276.

    Science.gov (United States)

    Drawz, Sarah M; Bethel, Christopher R; Hujer, Kristine M; Hurless, Kelly N; Distler, Anne M; Caselli, Emilia; Prati, Fabio; Bonomo, Robert A

    2009-06-02

    Inhibitor-resistant class A beta-lactamases of the TEM and SHV families that arise by single amino acid substitutions are a significant threat to the efficacy of beta-lactam/beta-lactamase inhibitor combinations. To better understand the basis of the inhibitor-resistant phenotype in SHV, we performed mutagenesis to examine the role of a second-shell residue, Asn276. Of the 19 variants expressed in Escherichia coli, only the Asn276Asp enzyme demonstrated reduced susceptibility to ampicillin/clavulanate (MIC increased from 50/2 --> 50/8 microg/mL) while maintaining high-level resistance to ampicillin (MIC = 8192 microg/mL). Steady-state kinetic analyses of Asn276Asp revealed slightly diminished k(cat)/K(m) for all substrates tested. In contrast, we observed a 5-fold increase in K(i) for clavulanate (7.4 +/- 0.9 microM for Asn276Asp vs 1.4 +/- 0.2 microM for SHV-1) and a 40% reduction in k(inact)/K(I) (0.013 +/- 0.002 microM(-1 )s(-1) for Asn276Asp vs 0.021 +/- 0.004 microM(-1) s(-1) for SHV-1). Timed electrospray ionization mass spectrometry of clavulanate-inhibited SHV-1 and SHV Asn276Asp showed nearly identical mass adducts, arguing for a similar pathway of inactivation. Molecular modeling shows that novel electrostatic interactions are formed between Arg244Neta2 and both 276AspOdelta1 and Odelta2; these new forces restrict the spatial position of Arg244, a residue important in the recognition of the C(3)/C(4) carboxylate of beta-lactam substrates and inhibitors. Testing the functional consequences of this interaction, we noted considerable free energy costs (+DeltaDeltaG) for substrates and inhibitors. A rigid carbapenem (meropenem) was most affected by the Asn276Asp substitution (46-fold increase in K(i) vs SHV-1). We conclude that residue 276 is an important second-shell residue in class A beta-lactamase-mediated resistance to substrates and inhibitors, and only Asn is able to precisely modulate the conformational flexibility of Arg244 required for successful

  15. 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.

  16. Population Genomic Analysis of 1,777 Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Isolates, Houston, Texas: Unexpected Abundance of Clonal Group 307.

    Science.gov (United States)

    Long, S Wesley; Olsen, Randall J; Eagar, Todd N; Beres, Stephen B; Zhao, Picheng; Davis, James J; Brettin, Thomas; Xia, Fangfang; Musser, James M

    2017-05-16

    Klebsiella pneumoniae is a major human pathogen responsible for high morbidity and mortality rates. The emergence and spread of strains resistant to multiple antimicrobial agents and documented large nosocomial outbreaks are especially concerning. To develop new therapeutic strategies for K. pneumoniae , it is imperative to understand the population genomic structure of strains causing human infections. To address this knowledge gap, we sequenced the genomes of 1,777 extended-spectrum beta-lactamase-producing K. pneumoniae strains cultured from patients in the 2,000-bed Houston Methodist Hospital system between September 2011 and May 2015, representing a comprehensive, population-based strain sample. Strains of largely uncharacterized clonal group 307 (CG307) caused more infections than those of well-studied epidemic CG258. Strains varied markedly in gene content and had an extensive array of small and very large plasmids, often containing antimicrobial resistance genes. Some patients with multiple strains cultured over time were infected with genetically distinct clones. We identified 15 strains expressing the New Delhi metallo-beta-lactamase 1 (NDM-1) enzyme that confers broad resistance to nearly all beta-lactam antibiotics. Transcriptome sequencing analysis of 10 phylogenetically diverse strains showed that the global transcriptome of each strain was unique and highly variable. Experimental mouse infection provided new information about immunological parameters of host-pathogen interaction. We exploited the large data set to develop whole-genome sequence-based classifiers that accurately predict clinical antimicrobial resistance for 12 of the 16 antibiotics tested. We conclude that analysis of large, comprehensive, population-based strain samples can assist understanding of the molecular diversity of these organisms and contribute to enhanced translational research. IMPORTANCE Klebsiella pneumoniae causes human infections that are increasingly difficult to

  17. Characterization of the Extended-Spectrum beta-Lactamase Producers among Non-Fermenting Gram-Negative Bacteria Isolated from Burnt Patients

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    Mojdeh Hakemi Vala

    2013-09-01

    Full Text Available Please cite this article as: Hakemi Vala M, Hallajzadeh M, Fallah F, Hashemi A, Goudarzi H. Characterization of the extended-spectrum beta-lactamase producers among non-fermenting gram-negative bacteria isolated from burnt patients. Arch Hyg Sci 2013;2(1:1-6. Background & Aims of the Study: Extended-spectrum beta-Lactamases (ESBLs represent a major group of beta-lactamases which are responsible for resistance to oxyimino-cephalosporins and aztreonam and currently being identified in large numbers throughout the world. The objective of this study was to characterize ESBL producers among non-fermenter gram-negative bacteria isolated from burnt patients. Materials & Methods: During April to July 2012, 75 non-fermenter gram-negative bacilli were isolated from 240 bacterial cultures collected from wounds of burnt patients admitted to the Burn Unit at Shahid Motahari Hospital (Tehran, Iran. Bacterial isolation and identification was done using standard methods. Antimicrobial susceptibility testing was performed by disk diffusion method for all strains against selected antibiotics and minimum inhibitory concentration was determined by microdilution test. The ability to produce ESBL was detected through double disk synergy test among candidate strains. Results: Of 75 non-fermenter isolates, 47 Pseudomonas aeruginosa and 28 Acinetobacter baumannii were identified. The resistance of P. aeruginosa isolates to tested antibiotics in antibiogram test were 100% to cefpodoxime, 82.98% to ceftriaxone, 78.73% to imipenem, 75% to meropenem, 72.72% to gentamicin, 69.23% to ciprofloxacin and aztreonam, 67.57% to cefepime, 65.95% to ceftazidime, and 61.53% to piperacillin. The results for Acinetobacter baumannii were 100% to ceftazidime, cefepime, ciprofloxacin, imipenem, meropenem, cefpodoxime, and cefotaxim, 96.85% to gentamicin, 89.65% to ceftriaxone, 65.51% to aztreonam, and 40% to piperacillin. Double disk synergy test showed that 21 (28% of non

  18. Prevalence of AmpC type extended spectrum beta lactamases genes in clinical Samples of E.coli Isolated from Poultry and Humans

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    Elham Farrokhnazar

    2016-07-01

    Full Text Available Emergence of antibiotic resistance among pathogens, particularly in health centers and hospitals, has become a major public health problem. This study identified AmpC-type beta-lactamase against the antibiotic ceftazidime, cefotaxime and cefpodoxime in E.coli isolated from human and poultry and types of producing genes were studied by PCR. In this study, 500 clinical human samples of urine from hospitals of Tehran during 5 months as well as 300 poultry samples were collected and transferred to the microbiology laboratory. Biochemical tests such as TSI, Urea and IMViC were performed on suspected colonies with E.coli. To identify ESBL producing strains, beta-lactamase samples were cultured on Mueller-Hinton agar through antimicrobial susceptibility test by disk agar diffusion based on the standard CLSI for initial screening. PCR reactions were done using specific primers CITM, EBCM, DHAM and MOXM to identify the beta-lactamase AmpC. A number of 200 human and 55 poultry E.coli samples were screened. In human samples, 105 (52.5% were resistant and potential producers of ESBL and AmpC; out of those, 102 (51% produced ESBL and 3 (1.5% potentially produced AmpC. In the study on 55 E.coli isolates from poultry samples based on the results of disk agar diffusion test, 4 (7.2% samples were resistant and potential producers of ESBL. None of the samples were AmpC producers. Through PCR, 2 human samples (1% were CITM positive and one sample (0.5% was DHAM positive. Through the PCR carried out on poultry samples, there were no bands with 4 primers. There was AmpC in human samples; while further studies are required for poultry samples, because poultry significantly contribute in production of food for humans and can be an important source for dissemination of antibiotic resistance. Given the significance of Ampc in providing high levels of beta-lactam antibiotic resistance, particularly third generation cephalosporins which are very common treatments, more

  19. Evaluation of the MicroScan ESBL plus confirmation panel for detection of extended-spectrum beta-lactamases in clinical isolates of oxyimino-cephalosporin-resistant Gram-negative bacteria.

    Science.gov (United States)

    Stürenburg, Enno; Lang, Melanie; Horstkotte, Matthias A; Laufs, Rainer; Mack, Dietrich

    2004-11-01

    We aimed to assess the performance of the MicroScan ESBL plus confirmation panel using a series of 87 oxyimino-cephalosporin-resistant Gram-negative bacilli of various species. Organisms tested included 57 extended-spectrum beta-lactamase (ESBL) strains comprising Enterobacter aerogenes (3), Enterobacter cloacae (10), Escherichia coli (11), Klebsiella pneumoniae (26), Klebsiella oxytoca (3) and Proteus mirabilis (4). Also included were 30 strains resistant to oxyimino cephalosporins but lacking ESBLs, which were characterized with other resistance mechanisms, such as inherent clavulanate susceptibility in Acinetobacter spp. (4), hyperproduction of AmpC enzyme in Citrobacter freundii (2), E. aerogenes (3), E. cloacae (3), E. coli (4), Hafnia alvei (1) and Morganella morganii (1), production of plasmid-mediated AmpC beta-lactamase in K. pneumoniae (3) and E. coli (3) or hyperproduction of K1 enzyme in K. oxytoca (6). The MicroScan MIC-based clavulanate synergy correctly classified 50 of 57 ESBL strains as ESBL-positive and 23 of 30 non-ESBL strains as ESBL-negative (yielding a sensitivity of 88% and a specificity of 76.7%, respectively). False negatives among ESBL producers were highest with Enterobacter spp. due to masking interactions between ESBL and AmpC beta-lactamases. False-positive classifications occurred in two Acinetobacter spp., one E. coli producing plasmid-mediated AmpC beta-lactamase and two K. oxytoca hyperproducing their chromosomal K1 beta-lactamase. The MicroScan clavulanate synergy test proved to be a valuable tool for ESBL confirmation. However, this test has limitations in detecting ESBLs in Enterobacter spp. and in discriminating ESBL-related resistance from the K1 enzyme and from inherent clavulanate susceptibility in Acinetobacter spp.

  20. 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

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    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

  1. 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%.

  2. 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.

  3. Plasmid-mediated AmpC beta-lactamase-producing Escherichia coli causing urinary tract infection in the Auckland community likely to be resistant to commonly prescribed antimicrobials.

    Science.gov (United States)

    Drinkovic, Dragana; Morris, Arthur J; Dyet, Kristin; Bakker, Sarah; Heffernan, Helen

    2015-03-13

    To estimate the prevalence and characterise plasmid-mediated AmpC beta-lactamase (PMACBL)- producing Escherichia coli in the Auckland community. All cefoxitin non-susceptible (NS) E. coli identified at the two Auckland community laboratories between 1 January and 31 August 2011 were referred to ESR for boronic acid double-disc synergy testing, to detect the production of AmpC beta-lactamase, and polymerase chain reaction (PCR) to identify the presence of PMACBL genes. PMACBL-producing isolates were typed using pulsed-field gel electrophoresis (PFGE), and PCR was used to determine their phylogenetic group and to identify multilocus sequence type (ST)131. Antimicrobial susceptibility testing and detection of extended-spectrum beta-lactamases (ESBLs) were performed according to the Clinical and Laboratory Standards Institute recommendations. 101 (51%) and 74 (37%) of 200 non-duplicate cefoxitin-NS E. coli were PMACBL producers or assumed hyper-producers of chromosomal AmpC beta-lactamase, respectively. The prevalence of PMACBL-producing E. coli was 0.4%. PMACBL-producing E. coli were significantly less susceptible to norfloxacin, trimethoprim and nitrofurantoin than E. coli that produced neither a PMACBL nor an ESBL. Very few (4%) PMACBL-producing E. coli co-produced an ESBL. Most (88%) of the PMACBL-producing isolates had a CMY-2-like PMACBL. The PMACBL-producing E. coli isolates were diverse based on their PFGE profiles, 44% belonged to phylogenetic group D, and only four were ST131. 100 of the 101 PMACBL-producing E. coli were cultured from urine, and were causing urinary tract infection (UTI) in the majority of patients. The median patient age was 56 years and most (94%) of the patients were women. A greater proportion of patients with community-acquired UTI caused by PMACBL-producing E. coli received a beta-lactam antimicrobial than patients with community-acquired UTI caused by other non-AmpC, non-ESBL-producing E. coli. Thirty-six (43%) patients with community

  4. Prevalence and antimicrobial susceptibility patterns of extended spectrum beta-lactamase producing Entrobacteriaceae in the University of Gondar Referral Hospital environments, northwest Ethiopia.

    Science.gov (United States)

    Engda, Tigist; Moges, Feleke; Gelaw, Aschalew; Eshete, Setegn; Mekonnen, Feleke

    2018-05-22

    This study aimed at assessing the magnitude, distribution, and the antimicrobial susceptibility of the extended spectrum beta-lactamase producing Entrobacteriaceae in the University of Gondar Referral Hospital environments. Out of a total of 384 samples, 14.8% were ESBL producing Entrobacteriaceae, where 42.10% Klebsiella pneumoniae, 35.09% Escherchia coli and 7.01% Proteus mirabilis were the predominant isolates. Most ESBL producing isolates, that is, 24.56, 22.8, and 22.8% were found from waste water, sinks and bedside tables respectively. All ESBL producing Entrobacteriaceae were found to be resistant to ceftriaxone, ceftazidime, cefpirome, cefpodoxime, and amoxicillin with Clavulanic acid. Resistance rate was also high for non-beta-lactam antimicrobials, like chloramphenicol (70.18%), cotrimoxazole (64.91%), norfloxacin (42.10%), ciprofloxacin (43.86%), and gentamicin (19.30%).

  5. Emergence of extended spectrum beta-lactamases-producing strains belonging to cefotaxime-M-1 class from intensive care units patients and environmental surfaces in Pakistan

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    Aqsa Ashraf Bukhari

    2016-11-01

    Full Text Available Aim: The emergence of multidrug-resistant (MDR bacteria is the most dangerous threat for the treatment of infectious diseases. The aim of this study was to detect and characterize extended spectrum beta-lactamases (ESBLs and carbapenemase-producing Klebsiella pneumoniae and Escherichia coli among patients and environment of intensive care units (ICUs of three tertiary care hospitals in Pakistan. Materials and Methods: A total of 82 samples from ICU’s patients and inanimate environment (injection trays, wash basins, door handles, hand swabs of professionals, and ICU fridges were screened for ESBL by culturing on CHROMagar-ESBL. ESBL and carbapenemases production were confirmed by double disc synergy test and modified Hodge’s test, respectively. Polymerase chain reaction was used to detect ESBL encoding genes bla cefotaxime (CTX-M, blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaTEM, blaSHV and carbapenemase genes blaKPC, bla New Delhi metallo-beta-lactamase-1, blaOXA-48 and blaVIM. Results: Overall, ESBL production was found high 30/82 (36.5% among isolates of which 15.8% K. pneumoniae and 20.7% E. coli were identified. All the K. pneumoniae and majority of E. coli isolates were MDR, i.e., resistance to three or more antimicrobial categories. Molecular characterization showed the blaCTX-M-1 as the predominant genotype found in 17/21 (80% of the isolates. None of the strains was found positive for carbapenemase-encoding genes. Conclusion: In conclusion, this study demonstrates the emergence of MDR ESBL producing strains among ICU patients and hospital environment, posing a serious threat for the control of nosocomial infections.

  6. Population Genomic Analysis of 1,777 Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Isolates, Houston, Texas: Unexpected Abundance of Clonal Group 307

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    S. Wesley Long

    2017-05-01

    Full Text Available Klebsiella pneumoniae is a major human pathogen responsible for high morbidity and mortality rates. The emergence and spread of strains resistant to multiple antimicrobial agents and documented large nosocomial outbreaks are especially concerning. To develop new therapeutic strategies for K. pneumoniae, it is imperative to understand the population genomic structure of strains causing human infections. To address this knowledge gap, we sequenced the genomes of 1,777 extended-spectrum beta-lactamase-producing K. pneumoniae strains cultured from patients in the 2,000-bed Houston Methodist Hospital system between September 2011 and May 2015, representing a comprehensive, population-based strain sample. Strains of largely uncharacterized clonal group 307 (CG307 caused more infections than those of well-studied epidemic CG258. Strains varied markedly in gene content and had an extensive array of small and very large plasmids, often containing antimicrobial resistance genes. Some patients with multiple strains cultured over time were infected with genetically distinct clones. We identified 15 strains expressing the New Delhi metallo-beta-lactamase 1 (NDM-1 enzyme that confers broad resistance to nearly all beta-lactam antibiotics. Transcriptome sequencing analysis of 10 phylogenetically diverse strains showed that the global transcriptome of each strain was unique and highly variable. Experimental mouse infection provided new information about immunological parameters of host-pathogen interaction. We exploited the large data set to develop whole-genome sequence-based classifiers that accurately predict clinical antimicrobial resistance for 12 of the 16 antibiotics tested. We conclude that analysis of large, comprehensive, population-based strain samples can assist understanding of the molecular diversity of these organisms and contribute to enhanced translational research.

  7. Occurrence of qnr-positive clinical isolates in Klebsiella pneumoniae producing ESBL or AmpC-type beta-lactamase from five pediatric hospitals in China.

    Science.gov (United States)

    Wang, Aihua; Yang, Yonghong; Lu, Quan; Wang, Yi; Chen, Yuan; Deng, Li; Ding, Hui; Deng, Qiulian; Wang, Li; Shen, Xuzhuang

    2008-06-01

    The plasmid-mediated quinolone resistance qnr genes in clinical isolates in adults have been described in different countries; however, the frequency of their occurrence has not been detected in pediatric patients. A total of 410 clinical isolates of Klebsiella pneumoniae, identified as producers of an extended-spectrum beta-lactamase (ESBL), or AmpC beta-lactamase, were collected from five children's hospitals in China during 2005-2006. The isolates were screened for the presence of the qnrA, qnrB, and qnrS genes, and then the harboring qnr gene isolates were detected for a bla gene coding for the TEM, SHV, CTX-M, and plasmid-mediated ampC gene by a PCR experiment. Ninety-two isolates (22.7%) were positive for the qnr gene, including 10 of qnrA (2.4%), 25 of qnrB (6.1%), and 62 of qnrS (15.1%). Eighty-one of the 92 (88.0%) qnr-positive isolates carried at least one bla gene for TEM, SHV, CTX-M, or DHA-1. The ciprofloxacin resistance increased 16-256-fold and oflaxacin resistance increased 2-32-fold in transconjugants, respectively. These results indicated that the plasmid-mediated qnr quinolone resistance gene was qnrS, followed by qnrB and qnrA. Most of the isolates also carried a bla gene coding ESBL or ampC gene coding DHA-1 among Klebsiella pneumoniae isolated from Chinese pediatric patients.

  8. Spatial molecular epidemiology of carbapenem-resistant and New Delhi metallo beta-lactamase (blaNDM)-producing Escherichia coli in the piglets of organized farms in India.

    Science.gov (United States)

    Pruthvishree, B S; Vinodh Kumar, O R; Sinha, D K; Malik, Y P S; Dubal, Z B; Desingu, P A; Shivakumar, M; Krishnaswamy, N; Singh, B R

    2017-06-01

    A cross-sectional study was conducted in 10 government-organized pig farms between 2014 and 2016 representing seven states of India to understand the epidemiology of carbapenem resistance in the Escherichia coli. In this study, fecal sample (n = 673) from non-diarrheic (n = 501) and diarrheic (n = 172) piglets were processed for isolation of carbapenem resistant E. coli. Of 673, E. coli isolate (n = 112) was genotyped for confirming the carbapenem resistance and associated virulence factors. Of the 112 isolates, 23 were phenotypically resistant to carbapenem and 8 were carrying the New Delhi metallo beta-lactamase (blaNDM) gene. The carbapenem-resistant isolates also produced extended spectrum beta-lactamases and were multidrug resistant. The PCR-based pathotyping revealed the presence of stx1, stx2, eae and hlyA genes. The enterobacterial repetitive intergenic consensus PCR dendrogram analysis of the isolates yielded three distinct clusters. The statistical analysis revealed no association between carriages of carbapenem-resistant E. coli in different breed of piglets however, location, sex, health status of piglets and age showed significant difference. The spatial analysis with SaTScan helped in identification of carbapenem-resistant clusters. The presence of carbapenem resistant E. coli isolates with virulence genes in the piglet poses a potential public health risk through possible access and spread via the food chain and environment. Efflux pump may also play an important role in carbapenem resistance in piglet E. coli isolates. Furthermore, identification of risk factors in relation to spatial clusters will help in designing preventive strategies for reducing the risk of spread of carbapenem resistant bacteria. 1. Piglets harbor carbapenem resistant E. coli and have great public health significance. 2. Apart from carbapenemase, efflux pump is also important for carbapenem resistance. 3. This is the first report of blaNDM in the piglets from India. © 2017

  9. 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.

  10. Caracterização dos Genes de metalo-beta-lactamase blaNDM, blaSPM e blaIMP em Pseudomonas aeruginosa Resistentes a Carbapenens

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    Carolina Jahn

    2017-01-01

    Full Text Available Justificativa e objetivo: P. aeruginosa é responsável por causar grande variedade de infecções agudas e crônicas. A resistência aos carbapenêmicos está se tornando um problema terapêutico mundial e a produção de metalo-beta-lactamases (MBL tem surgido como um dos mecanismos responsáveis por esta resistência. Objetivou-se identificar as cepas produtoras de MBL e verificar a presença dos genes codificador blaNDM, blaSPM e blaIMP em isolados de P. aeruginosa resistentes aos carbapenens. Métodos: Foram analisadas 16 cepas de P. aeruginosa, isoladas de diferentes sítios de indivíduos atendidos pelo Hospital do Vale do Rio Pardo-RS, no período de março 2014 a fevereiro de 2015. A identificação bioquímica foi realizada segundo protocolo da ANVISA e o antibiograma pelo método de ágar difusão em disco. A triagem fenotípica foi feita pelo teste modificado de aproximação de discos utilizando o EDTA como inibidor e a presença dos genes através da técnica de reação em cadeia da polimerase. Resultados: O setor clínico com maior prevalência de isolamento de P. aeruginosa resistente aos carbapenens é a unidade de terapia intensiva (50%. Em relação aos espécimes clínicos, 62,5% dos isolados foram identificados no trato respiratório inferior. Através do teste fenotípico, 50% das amostras analisadas mostraram ser produtora da enzima MBL. Através da técnica de reação em cadeia da polimerase foi possível identificar a presença de um dos três genes propostos, sendo eles blaSPM. Conclusão: Os isolados de P. aeruginosa mostraram serem produtores de MBL e apresentaram o gene de resistência blaSPM nesta espécie no interior do Rio Grande do Sul. Palavras Chaves: Pseudomonas aeruginosa; metalo-beta-lactamases; blaNDM, blaIMP, e blaSPM .

  11. 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.

  12. 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)

    Manikprabhu, Deene; Lingappa, K.

    2014-01-01

    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

  13. Looking for the new preparations for antibacterial therapy. II. Clinical trials; new beta-lactam antibiotics and beta-lactamase inhibitors.

    Science.gov (United States)

    Karpiuk, Izabela; Tyski, Stefan

    2013-01-01

    To obtain a status of a medicinal product, a compound possessing potential antimicrobial activity and displaying no cytotoxicity, must undergo three phases of clinical trials to prove its therapeutic efficacy, safety and quality. Properties of the compound should be based on the results of studies meeting specific criteria. Studies should be: randomized, double-blind, involving sufficient number of volunteers, concerning the infections localized in strictly defined area and caused by identified microorganisms. After the medicinal product is authorized to be on the market, clinical trials of the fourth phase are carried out to detect adverse effects, overdose symptoms, interactions of the new drug with other medicinal products and to establish characteristic of activity among groups such as children, elderly, women in pregnancy and patients suffering from other diseases, but only if the benefits of receiving treatment outweigh the risks. This article is a second part of the series associated with searching for new antibacterial agents and it relates to performance of clinical trials and the new compounds belonging to the class of beta-lactams. Among the 9 presented compounds, candidates to become medicinal products, two belong to the cephalosporins (CXA-101, S-649266), one to carbapenems (razupenem), three to monobactams (BAL30072, BAL30376, MC-1) and three to beta-lactamase inhibitors (NXL-104, MK-7655, ME1071).

  14. 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

    2000-01-01

    The aim of this study was to examine the development of resistance of biofilm-growing P. aeruginosa during treatment with ceftazidime. Biofilms were established in vitro using a modified Robbins device (MRD) and in vivo in the rat model of chronic lung infection. Three P. aeruginosa strains...... of ceftazidime to biofilms established in MDR, a statistically significant development of resistance to ceftazidime in PAO 579 or 19676A bacterial populations occurred. When ceftazidime was administered 4 h/day (200 mg/l) for 2 weeks, the frequency of resistant 19676A having MIC>25 mg/l was 4.4 10(-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 579...

  15. Empiric Piperacillin-Tazobactam versus Carbapenems in the Treatment of Bacteraemia Due to Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae.

    Science.gov (United States)

    Ng, Tat Ming; Khong, Wendy X; 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%]), Pcarbapenem.

  16. Detection of Extended Spectrum Beta-Lactamases Resistance Genes among Bacteria Isolated from Selected Drinking Water Distribution Channels in Southwestern Nigeria.

    Science.gov (United States)

    Adesoji, Ayodele T; Ogunjobi, Adeniyi A

    2016-01-01

    Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include bla TEM, bla SHV, and bla CTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with bla TEM being the most abundant (50/61) and bla CTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, bla SHV + bla TEM or bla CTX + bla TEM, were observed in 11 and 1 isolate, respectively. In conclusion, classic bla TEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria.

  17. OXA-48 and CTX-M-15 extended-spectrum beta-lactamases in raw milk in Lebanon: epidemic spread of dominant Klebsiella pneumoniae clones.

    Science.gov (United States)

    Diab, Mohamad; Hamze, Monzer; Bonnet, Richard; Saras, Estelle; Madec, Jean-Yves; Haenni, Marisa

    2017-11-01

    Raw milk has recently been reported as a source of extended-spectrum beta-lactamase (ESBL) and carbapenemase genes. We thus investigated the prevalence of ESBL- and carbapenemase-producing Enterobacteriaceae in raw milk in Lebanon in order to assess the risk of transfer of these bacteria to humans. A high prevalence (30.2 %) of CTX-M-15-producing K. pneumoniae was detected in raw bovine milk. Three main K. pneumoniae clones were identified by PFGE and MLST typing. Southern blot experiments revealed that one of these clones carried the blaCTX-M-15 gene chromosomally. Moreover, one OXA-48-producing K. pneumoniae ST530 and seven CTX-M-15-producing Escherichia coli sharing the same ST were also detected. These findings highlight the spread of dominant CTX-M-15-producing K. pneumoniae clones and OXA-48-producing isolates in the food chain. Milk, which is mostly consumed raw in Lebanon, may be a source of human exposure to ESBLs and carbapenemases.

  18. Prostatic Abscess after Stapled Hemorrhoidopexy Caused by ESBL Extended Spectrum Beta Lactamase Producing Klebsiella pneumoniae: An Additional Challenge to Postoperative Sepsis

    Directory of Open Access Journals (Sweden)

    Asem Saleh

    2017-01-01

    Full Text Available Postoperative septic complications of hemorrhoids surgical interventions are rare, but very serious with high mortality rate. Early diagnosis and prompt therapy are essential to save patient’s life. There are a good number of articles and case reports about these septic complications. We are presenting a case report of a prostatic abscess caused by extended spectrum beta lactamase (ESBL producing Klebsiella pneumoniae after hemorrhoidopexy. Our patient was a healthy middle aged Saudi male who has no significant medical history apart from morbid obesity and recurrent urinary tract infections. ESBL producing K. pneumoniae could be detected only after aspiration of the prostatic abscess, but proper antibiotic was introduced intravenously on admission before culture of aspirate of the abscess was available. Antibiotic was continued for 30 days and abscess resolved completely. In our electronic search, we could not find any case report of prostatic abscess after stapled hemorrhoidopexy caused by ESBL producing organism. This is an additional challenge for treating physicians as these organisms are sensitive only to one group of antibiotics (carbapenem group.

  19. Antibiotic Susceptibilities and Genetic Characteristics of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Isolates from Stools of Pediatric Diarrhea Patients in Surabaya, Indonesia.

    Science.gov (United States)

    Bagus Wasito, Eddy; Shigemura, Katsumi; Osawa, Kayo; Fardah, Alpha; Kanaida, Akiho; Raharjo, Dadik; Kuntaman, K; Hadi, Usman; Harijono, Sugeng; Marto Sudarmo, Subijanto; Nakamura, Tatsuya; Shibayama, Keigo; Fujisawa, Masato; Shirakawa, Toshiro

    2017-07-24

    The purpose of this study was to investigate extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates from pediatric (aged 0 to 3 years) diarrhea patients in Surabaya, Indonesia, where this kind of survey is rare; our study included assessment of their antibiotic susceptibilities, as well as ESBL typing, multilocus sequence typing (MLST), and diarrheagenic E. coli (DEC)-typing. ESBL-producing E. coli were detected in 18.8% of all the samples. Many ESBL-producing E. coli had significantly lower susceptibility to gentamicin (p < 0.0001) and the quinolones nalidixic acid (p=0.004) and ciprofloxacin (p < 0.0001) than non-producers. In ESBL-producing E. coli, 84.0% of strains expressed CTX-M-15 alone or in combination with other ESBL types. MLST revealed that 24.0% of ESBL-producers had sequence type 617, all of which expressed the CTX-M-15 gene; we also detected expression of 3 DEC-related genes: 2 enteroaggregative E. coli genes and 1 enteropathogenic E. coli gene. In conclusion, CTX-M-15-type ESBL-producing E. coli ST617 appear to have spread to Indonesia.

  20. Validation of Minim typing for fast and accurate discrimination of extended-spectrum, beta-lactamase-producing Klebsiella pneumoniae isolates in tertiary care hospital.

    Science.gov (United States)

    Brhelova, Eva; Kocmanova, Iva; Racil, Zdenek; Hanslianova, Marketa; Antonova, Mariya; Mayer, Jiri; Lengerova, Martina

    2016-09-01

    Minim typing is derived from the multi-locus sequence typing (MLST). It targets the same genes, but sequencing is replaced by high resolution melt analysis. Typing can be performed by analysing six loci (6MelT), four loci (4MelT) or using data from four loci plus sequencing the tonB gene (HybridMelT). The aim of this study was to evaluate Minim typing to discriminate extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KLPN) isolates at our hospital. In total, 380 isolates were analyzed. The obtained alleles were assigned according to both the 6MelT and 4MelT typing scheme. In 97 isolates, the tonB gene was sequenced to enable HybridMelT typing. We found that the presented method is suitable to quickly monitor isolates of ESBL-KLPN; results are obtained in less than 2 hours and at a lower cost than MLST. We identified a local ESBL-KLPN outbreak and a comparison of colonizing and invasive isolates revealed a long term colonization of patients with the same strain. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. 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.

  2. 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.

  3. Escherichia coli-producing extended-spectrum beta-lactamase CTX-M-15 in a captive South American tapir (Tapirus terrestris).

    Science.gov (United States)

    Klimes, Jiri; Machalkova, Marketa; Dolejska, Monika; Cizek, Alois; Janoszowska, Dagmar; Alexa, Pavel; Albrechtova, Katerina; Vojtech, Jiri; Literak, Ivan

    2013-03-01

    Only a few reports exist on the occurrence of resistant bacteria in zoo animals. Therefore, an isolation of multiresistant Escherichia coli from the lungs of a captive South American tapir (Tapirus terrestris) lead to its characterization and further investigation of samples from animals inhabiting the same paddock and from the shared environment. The tapir suffered from an intermandibular abscess and pneumonia and was euthanatized after unsuccessful therapy, including administration of antibiotics. The authors performed selective isolation of extended-spectrum beta-lactamase (ESBL)-positive E. coli strains and identification of resistance genes using polymerase chain reaction. Seven multiresistant, ESBL-producing E. coli isolates were obtained, all belonging to the B2 phylogenetic group and showing identical profile on pulsed-field gel electrophoresis. These isolates carried several resistance genes, including the gene bla(CTX-M-15). This case demonstrates the transmission of related epidemiologically important E. coli isolates whose potential transmission to other animals and zoo staff can be assumed.

  4. Efficacy of a once-a-week screening programme to control extended-spectrum beta-lactamase-producing bacteria in a neonatal intensive care unit.

    Science.gov (United States)

    Rybczynska, Helena; Melander, Eva; Johansson, Hugo; Lundberg, Fredrik

    2014-06-01

    Extended-spectrum beta-lactamase (ESBL)-producing bacteria are an escalating problem threatening health. Devastating consequences can result in neonatal intensive care units (NICU) due to these bacteria. The aim of this study was to investigate the efficacy of once-a-week screening (July 2010 to September 2012) versus screening on demand (April 2008 to June 2010). The investigation was an open retrospective descriptive study comparing 2 unpaired groups, the first exposed to screening on demand and the second to screening once a week. All other infection control measures were unchanged. Both groups were cared for in the NICU of Skåne University Hospital. Parameters compared were the proportion of cultured neonates, prevalence, time before detection, number of secondary cases, and clinical infections due to ESBL-producing bacteria. The proportion of cultured neonates increased from 28% to 49% (p control the epidemiology of unwanted pathogens among newborn infants. It provides the opportunity for early intervention, thereby avoiding secondary cases and infections. Premature neonates in particular benefit from this approach. The prevalence of ESBL of 1.77% is low from an international perspective. ESBL appear to be introduced onto the ward by mothers colonized with ESBL.

  5. 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.

  6. 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.

  7. A passage from India: Association between air traffic and reported cases of New Delhi Metallo-beta-lactamase 1 from 2007 to 2012.

    Science.gov (United States)

    MacFadden, Derek R; Bogoch, Isaac I; Brownstein, John S; Daneman, Nick; Fisman, David; German, Matthew; Khan, Kamran

    2015-01-01

    Highly transmissible genes encoding resistance to carbapenems have demonstrated global spread. The New Delhi Metallo-beta-lactamase 1 gene is hypothesized to have originated in India, with subsequent dissemination by colonized or infected travelers. We conducted an ecological study evaluating the association between the cumulative air traffic departing India between 2007 and 2012 and published cases of NDM-1. Receiver operator characteristic curves were generated as well as multivariate logistic regression models. 193 countries with complete flight and World Bank data were included in the analysis. Receiver operator characteristic curves (ROC) of the dichotomous outcome of a published case of NDM-1 were generated, yielding an unadjusted area under the curve (AUC) of 0.88 and adjusted AUC of 0.85. The unadjusted odds ratio of having a reported case of NDM-1, for every percentage increase in cumulative air traffic departing India, was 2.3 (95% CI 1.4 to 3.7) and adjusted was 2.0 (95% CI 1.2 to 3.4). We demonstrate that flows of international travelers departing India by air is associated with published NDM-1 cases, globally. Countries with high passenger flight traffic from India with no reported cases of NDM-1 may be at increased risk of having unreported transmission of NDM-1. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Antimicrobial susceptibility and molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacteriaceae from intensive care units at Hamad Medical Corporation, Qatar.

    Science.gov (United States)

    Sid Ahmed, Mazen A; Bansal, Devendra; Acharya, Anushree; Elmi, Asha A; Hamid, Jemal M; Sid Ahmed, Abuelhassan M; Chandra, Prem; Ibrahim, Emad; Sultan, Ali A; Doiphode, Sanjay; Bilal, Naser Eldin; Deshmukh, Anand

    2016-01-01

    The emergence of extended-spectrum beta-lactamase (ESBL)-producing isolates has important clinical and therapeutic implications. High prevalence of ESBL-producing Enterobacteriaceae has been reported in the literature for clinical samples from a variety of infection sites. The present study was undertaken to evaluate the prevalence of ESBL-producing Enterobacteriaceae, and to perform molecular characterization and antimicrobial susceptibility testing of clinical isolates from patients admitted to the intensive care units at Hamad Medical Corporation, Doha, Qatar, from November 2012 to October 2013. A total of 629 Enterobacteriaceae isolates were included in the study. Identification and susceptibility testing was performed using Phoenix (Becton Dickinson) and the ESBL producers were confirmed by double-disk potentiation as recommended by the Clinical and Laboratory Standards Institute. Molecular analysis of the ESBL producers was performed by polymerase chain reaction. In total, 109 isolates (17.3 %) were confirmed as ESBL producers and all were sensitive to meropenem in routine susceptibility assays. Most of the ESBL producers (99.1 %) were resistant to amoxicillin/clavulanic acid and ceftriaxone and 93.6 % were resistant to cefepime. Among the ESBL-producing genes, bla CTX-M (66.1 %) was the most prevalent, followed by bla SHV (53.2 %) and bla TEM (40.4 %). These findings show the high prevalence of ESBL-producing Enterobacteriaceae within the intensive care units at Hamad Medical Corporation, Qatar, and emphasize the need for judicious use of antibiotics and the implementation of strict infection control measures.

  9. Transcriptional Alterations of Virulence-Associated Genes in Extended Spectrum Beta-Lactamase (ESBL-Producing Uropathogenic Escherichia coli during Morphologic Transitions Induced by Ineffective Antibiotics

    Directory of Open Access Journals (Sweden)

    Isak Demirel

    2017-06-01

    Full Text Available It is known that an ineffective antibiotic treatment can induce morphological shifts in uropathogenic Escherichia coli (UPEC but the virulence properties during these shifts remain to be studied. The present study examines changes in global gene expression patterns and in virulence factor-associated genes in an extended spectrum beta-lactamase (ESBL-producing UPEC (ESBL019 during the morphologic transitions induced by an ineffective antibiotic and in the presence of human primary bladder epithelial cells. Microarray results showed that the different morphological states of ESBL019 had significant transcriptional alterations of a large number of genes (Transition; 7%, Filamentation; 32%, and Reverted 19% of the entities on the array. All three morphological states of ESBL019 were associated with a decreased energy metabolism, altered iron acquisition systems and altered adhesion expression. In addition, genes associated with LPS synthesis and bacterial motility was also altered in all the morphological states. Furthermore, the transition state induced a significantly higher release of TNF-α from bladder epithelial cells compared to all other morphologies, while the reverted state was unable to induce TNF-α release. Our findings show that the morphological shifts induced by ineffective antibiotics are associated with significant transcriptional virulence alterations in ESBL-producing UPEC, which may affect survival and persistence in the urinary tract.

  10. A trial with IgY chicken antibodies to eradicate faecal carriage of Klebsiella pneumoniae and Escherichia coli producing extended-spectrum beta-lactamases

    Directory of Open Access Journals (Sweden)

    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.

  11. Frequency of Extended-Spectrum Beta-lactamases (ESBLs) in strains of Klebsiella and E. coli isolated from patients hospitalized in Yazd.

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    Zandi, Hengameh; Tabatabaei, Seyed Mostafa; Ehsani, Fatemeh; Zarch, Mojtaba Babaei; Doosthosseini, Samira

    2017-02-01

    Frequency of extended-spectrum beta-lactamases (ESBLs) and its variants may vary in different geographical areas, as reports indicate their spread in some certain communities. The aim of this study was to determine the frequency of ESBLs in strains of Klebsiella and E. coli , isolated from patients hospitalized in teaching hospitals of Yazd. This cross-sectional study was carried out on samples including E. coli and Klebsiella strains collected from laboratories of Shahid Sadoughi and Shahid Rahnemoun hospitals in Yazd, Iran in the period of 2011-2012. The colonies which were positive in lactose Eosin methylene-blue (EMB) medium were identified by biochemical methods, and 270 strains of Klebsiella and E. coli were isolated. Collected data and information were analyzed using Fisher's exact test and descriptive statistics such as mean in SPSS software, version 15, at a significant level of 0.05. In this study, 270 samples were examined, including 152 samples of E. coli (56.3%) and 118 samples of Klebsiella pneumonia (43.7%). Among the 152 samples of E. coli , 45 strains (30%) were producers of ESBLs. In addition, among the 118 samples of Klebsiella pneumonia , 44 strains (37.3%) were producers of ESBLs. E. coli strains showed the most resistance to Cefotaxime (100%), Ceftazidime (97.7%), and Cefepime (75.5%) respectively and Klebsiella strains showed the most resistance to Cefotaxime (100%), Ceftazidime (100%) and Cefepime (79.5%), respectively. Frequency of ESBLs in Klebsiella strains was higher than E. coli strains. No significant relationship was found between frequency of ESBLs and age or gender. In addition, E. coli strains showed the highest sensitivity to Imipenem, Amoxicillin/clavulanate, and Ciprofloxacin, while the highest antibiotic sensitivity of Klebsiella strains was shown to be to Piperacillin, Imipenem, and Amoxicillin/clavulanate.

  12. Maternal colonization or infection with extended-spectrum beta-lactamase-producing Enterobacteriaceae in Africa: A systematic review and meta-analysis.

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    Bulabula, Andre N H; Dramowski, Angela; Mehtar, Shaheen

    2017-11-01

    To summarize published studies on the prevalence of and risk factors for maternal bacterial colonization and/or infection with extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) in pregnant and/or post-partum women in Africa. A systematic review was conducted using the PubMed, Scopus, and Google Scholar databases. Bibliographies of included eligible studies were manually searched to identify additional relevant articles. No language restriction was applied. The timeframe of the search included all records from electronic database inception to July 15, 2017. A random-effects meta-analysis was performed to summarize the prevalence and the 95% confidence intervals (CI) of ESBL-E colonization or infection in pregnant or post-partum women in Africa. The meta-analysis was conducted using STATA IC 13.1 software and the metaprop function/plugin. Ten studies (seven on pregnant women and three on post-partum women) were included, documenting a 17% prevalence of maternal colonization with ESBL-E in Africa (95% CI 10-23%). The prevalence of ESBL-E in community isolates exceeded that in isolates from the hospital setting (22% vs. 14%). The most frequently reported ESBL-encoding gene was CTX-M (cefotaxime hydrolyzing capabilities). Data on risk factors for maternal ESBL-E colonization and infection are very limited. The prevalence of colonization and/or infection with ESBL-E in pregnant and post-partum women in Africa exceeds that reported from high- and middle-income settings, representing a risk for subsequent neonatal colonization and/or infection with ESBL-E. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  13. Clinical and economic outcomes associated with community-acquired intra-abdominal infections caused by extended spectrum beta-lactamase (ESBL) producing bacteria in China.

    Science.gov (United States)

    Hu, Bijie; Ye, Huifeng; Xu, Yingchun; Ni, Yuxing; Hu, Yunjian; Yu, Yunsong; Huang, Zhenfei; Ma, Larry

    2010-06-01

    To compare clinical and economic outcomes in patients with community-acquired intra-abdominal infection (IAI) due to extended spectrum beta-lactamase (ESBL) producing (ESBL-positive) bacteria versus non-ESBL-producing (ESBL-negative) bacteria in China. This was a retrospective chart review study of patients hospitalized with community-acquired IAI due to ESBL-positive or ESBL-negative infections caused by Escherichia coli or Klebsiella spp. Data were collected from six hospitals in China that participated in the Study for Monitoring Antibiotic Resistance Trends (SMART) during 2006-2007. Outcomes included clinical response at discharge and following first-line antibiotic, number of antibiotic agents and classes, duration of hospitalization, and overall hospitalization and intravenous antibiotic costs. Of the 85 patients included in the study, 32 (37.6%) had ESBL-positive and 53 (62.4%) had ESBL-negative infections; E. coli was responsible for 77.6% of infections. Infection resolved at discharge in 30 (93.8%) ESBL-positive and 48 (90.6%) ESBL-negative patients (P = NS). Fewer ESBL-positive patients achieved complete response following first-line antibiotics (56.3% versus 83.0%; P = 0.01). ESBL-positive patients required longer antibiotic treatment, more antibiotics, longer hospitalization (24.3 versus 14.6 days; 1.67-fold ratio; P = 0.001), and incurred higher hospitalization costs ( yen24,604 vs. yen13,788; $3604 vs. $2020; 1.78-fold ratio; P < 0.001). Patients with ESBL-positive infection had similar resolution rates at discharge compared to those with ESBL-negative infection, despite poorer first-line antibiotic response. However, ESBL-positive infection led to significantly greater hospitalization cost and intravenous antibiotic cost, and longer hospital stay.

  14. Novel sequence types of extended-spectrum and acquired AmpC beta-lactamase producing Escherichia coli and Escherichia clade V isolated from wild mammals.

    Science.gov (United States)

    Alonso, Carla Andrea; Alcalá, Leticia; Simón, Carmen; Torres, Carmen

    2017-08-01

    The closer contact with wildlife due to the growing human population and the destruction of natural habitats emphasizes the need of gaining insight into the role of animals as source of antimicrobial resistance. Here, we aim at characterizing the antimicrobial resistance genes and phylogenetic distribution of commensal Escherichia coli from 62 wild mammals. Isolates exhibiting resistance to ≥1 antibiotic were detected in 25.8% of the animals and 6.4% carried an extended-spectrum beta-lactamase (ESBL)/AmpC-producing E. coli. Genetic mechanisms involved in third-generation cephalosporin resistance were as follows: (i) hyperproduction of chromosomal AmpC (hedgehog), (ii) production of acquired CMY-2 β-lactamase (hedgehog), (iii) production of SHV-12 and CTX-M-14 ESBLs (n = 2, mink and roe-deer). ESBL genes were transferable by conjugation, and blaCMY-2 was mobilized by a 95kb IncI1 plasmid. The distribution of the phylogenetic groups in the E. coli collection studied was B1 (44.6%), B2 (24.6%), E (15.4%), A (4.6%) and F (3.1%). Five isolates (7.7%) were cryptic Escherichia clades (clade IV, 4 mice; clade V, 1 mink). ESBL/AmpC-E. coli isolates showed different sequence types (STs): ST1128/B1, ST4564/B1 (new), ST4996/B1 (new) and a non-registered ST. This study contributes to better understand the E. coli population and antimicrobial resistance flow in wildlife and reports new AmpC-E. coli STs and a first described ESBL-producing Escherichia clade V isolate. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Frequent use of colistin-based drug treatment to eliminate extended-spectrum beta-lactamase-producing Escherichia coli in backyard chicken farms in Thai Binh Province, Vietnam.

    Science.gov (United States)

    Nakayama, Tatsuya; Jinnai, Michio; Kawahara, Ryuji; Diep, Khong Thi; Thang, Nguyen Nam; Hoa, Tran Thi; Hanh, Le Kieu; Khai, Pham Ngoc; Sumimura, Yoshinori; Yamamoto, Yoshimasa

    2017-01-01

    Reports of livestock infections with extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-E) are increasing. Based on interviews conducted over a 6-month period, we found that veterinarians in the Vietnamese province of Thai Binh prefer to prescribe colistin-based drugs (CBD) in chicken farms. We aimed to clarify whether CBD use selects for strains of colistin-resistant ESBL-E. With the cooperation of seven local households, we detected ESBL-E in chickens' feces after treating chickens with CBD. Phylogenetic groupings and the presence of CTX-M/AmpC genes were determined, and the multi-antibiotic susceptibility of isolates was analyzed. Our results showed that ESBL-E presented in seven chickens' feces from two households. Seventy-two percent of ESBL-E isolates harbored CTX-M9 and the phylogenetic group A; the colistin minimum inhibitory concentration (MIC) of all isolated ESBL-E ranged from 0.064 to 1 μg mL -1 . Moreover, ESBL-E isolates were used to experimentally select for colistin resistance, and the effect of commercial CBD on ESBL-E was investigated. The results showed that an ESBL-E strain with a colistin MIC of 4 μg mL -1 was able to grow in media with CBD. Although CBD treatment was effective, in vitro experiments demonstrated that ESBL-E can easily acquire colistin resistance. Therefore, restrictions on colistin use are necessary to prevent the emergence of colistin-resistant bacteria.

  16. Determination of antimicrobial resistance pattern and Extended-Spectrum Beta Lactamases producing Pseudomonas aeruginosa strains isolated from clinical specimens of Hajar and Kashani Hospitals,Shahrekord 1387

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    Mana Shojapour

    2011-09-01

    Full Text Available Background: Pseudomonas aeruginosa is one of the leading causes of hospital infections in patients hospitalized for a 10 day period or over. It is also considered to be the most important cause of the burn wound infection. Approximately 75% of deaths in burned patients are due to wound infection and the subsequent septicemia. Clinical use of antibiotics has increasingly led to the global distribution of P. aeruginosa isolates with multi-drug resistance. The study was launched to determine the antimicrobial susceptibility pattern and the presence of the extended-spectrum-beta lactamase (ESBL in P.aeruginosa strains isolated from clinical specimens. Methods: Totally, 175 P. aeruginosa strains were isolated from clinical samples and identified by standard methods. The pattern of antimicrobial resistance was then performed on the isolates using Disk Agar Diffusion (DAD according to CLSI Guideline. Primary screening test for ESBL producing strains was performed by ceftazidim antibiotic disk using disk diffusion method. Combined disk method was used to confirm ESBL producing bacteria. Results: The rate of antimicrobial resistance of P.aeruginosa isolates were 64% to ticarcillin, 52.2% to cefepime, 68.6% to ticarcillin/clavolanic acid, 68.6% to ceftazidime, 67.4% to amikacin, 68.6% to gentamicin, 48% to imipenem, 77.7% to ciprofloxacin and 5.1% to polymixcine B. In the primary screening test, 120 isolates of P.aeruginosa strains were resistant to ceftazidime. In the combined disk method, 66 isolates (55% were positive for ESBLs. Conclusion: Polymixcine B was found to be the most effective antimicrobial agent in this study. Bacteria carrying ESBL genes may increase mortality and morbidity. Thus, their accurate diagnosis is of extreme importance to prevent from the treatment failure resulted from improper antibiotic administration.

  17. Anti-bacterial Efficacy of Bacteriocin Produced by Marine Bacillus subtilis Against Clinically Important Extended Spectrum Beta-Lactamase Strains and Methicillin-Resistant Staphylococcus aureus

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    Suresh Mickymaray

    2018-02-01

    Full Text Available Objective: To investigate the anti-bacterial efficacy of bacteriocin produced by Bacillus subtilis SM01 (GenBank accession no: KY612347, a Gram-positive marine bacterium, against Extended Spectrum Beta-Lactamase (ESBL producing Gram-negative pathogens Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli, and Gram-positive pathogen Methicillin-Resistant Staphylococcus aureus (MRSA. Methods: A marine bacterium was isolated from mangrove sediment from the Red Sea coast of Jeddah, Kingdom of Saudi Arabia, and identified based on its morphological, biochemical, and molecular characteristics. The bacteriocin production using this isolate was carried out in brain heart infusion broth (BHIB medium. The Anti-bacterial activity of bacteriocin was evaluated against selected ESBL strains and MRSA by the well agar method. The effects of incubation time, pH, and temperature on the Anti-bacterial activity were studied. Results: The bacteriocin Bac-SM01 produced by B. subtilis SM01 demonstrated broad-spectrum Anti-bacterial activity against both Gram-negative and -positive bacteria. The present study is the first report that the bacteriocin Bac-SM01 inhibits the growth of ESBL producing Gram-negative strains A. baumannii, P. aeruginosa, and E. coli, and a Gram-positive MRSA strain. The optimum incubation time, pH, and temperature for the Anti-bacterial activity of Bac-SM01 was 24 h, 7, and 37°C respectively. Conclusion: The overall investigation can conclude that the bacteriocin Bac-SM01 from the marine isolate Bacillus subtilis SM01 could be used as an alternative Anti-bacterial agent in pharmaceutical products.

  18. Detection of Healthcare-Related Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Transmission Events Using Combined Genetic and Phenotypic Epidemiology.

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    Anne F Voor In 't Holt

    Full Text Available Since the year 2000 there has been a sharp increase in the prevalence of healthcare-related infections caused by extended-spectrum beta-lactamase (ESBL-producing Escherichia coli. However, the high community prevalence of ESBL-producing E. coli isolates means that many E. coli typing techniques may not be suitable for detecting E. coli transmission events. Therefore, we investigated if High-throughput MultiLocus Sequence Typing (HiMLST and/or Raman spectroscopy were suitable techniques for detecting recent E. coli transmission events.This study was conducted from January until December 2010 at Erasmus University Medical Center, Rotterdam, the Netherlands. Isolates were typed using HiMLST and Raman spectroscopy. A genetic cluster was defined as two or more patients carrying identical isolates. We used predefined definitions for epidemiological relatedness to assess healthcare-related transmission.We included 194 patients; strains of 112 patients were typed using HiMLST and strains of 194 patients were typed using Raman spectroscopy. Raman spectroscopy identified 16 clusters while HiMLST identified 10 clusters. However, no healthcare-related transmission events were detected. When combining data from both typing techniques, we identified eight clusters (n = 34 patients, as well as 78 patients with a non-cluster isolate. However, we could not detect any healthcare-related transmission in these 8 clusters.Although clusters were genetically detected using HiMLST and Raman spectroscopy, no definite epidemiological relationships could be demonstrated which makes the possibility of healthcare-related transmission events highly unlikely. Our results suggest that typing of ESBL-producing E. coli using HiMLST and/or Raman spectroscopy is not helpful in detecting E. coli healthcare-related transmission events.

  19. Randomized controlled trial of piperacillin-tazobactam, cefepime and ertapenem for the treatment of urinary tract infection caused by extended-spectrum beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Seo, Yu Bin; Lee, Jacob; Kim, Young Keun; Lee, Seung Soon; Lee, Jeong-A; Kim, Hyo Youl; Uh, Young; Kim, Han-Sung; Song, Wonkeun

    2017-06-07

    Due to limited therapeutic options, the spread of extended-spectrum beta-lactamases (ESBLs) have become a major public health concern. We conducted a prospective, randomized, open-label comparison of the therapeutic efficacy of piperacillin-tazobactam (PTZ), cefepime, and ertapenem in febrile nosocomial urinary tract infection with ESBL-producing Escherichia coli (ESBL-EC). This study was conducted at three university hospitals between January 2013 and August 2015. Hospitalized adult patients presenting with fever were screened for healthcare-associated urinary tract infection (HA-UTI). When ESBL-EC was solely detected and susceptible to a randomized antibiotic in vitro, the case was included in the final analysis. Participants were treated for 10-14 days with PTZ, cefepime, or ertapenem. A total of 66 participants were evenly assigned to the PTZ and ertapenem treatment groups. After the recruitment of six participants, assignment to the cefepime treatment group was stopped because of an unexpectedly high treatment failure rate. The baseline characteristics of these participants did not differ from participants in other treatment groups. The clinical and microbiological response to PTZ treatment was estimated to be 94% and was similar to the response to ertapenem treatment. The efficacy of cefepime was 33.3%. In the cefepime group, age, Charlson comorbidity index, genotype, and minimal inhibitory concentration (MIC) did not significantly affect the success of treatment. Similarly, genotype seemed to be irrelevant with respect to clinical outcome in the PTZ group. Expired cases tended to involve septic shock with a high Charlson comorbidity index and high MIC. Results from this study suggest that PTZ is effective in the treatment of urinary tract infection caused by ESBL-EC when the in vitro test indicates susceptibility. In addition, cefepime should not be used as an alternative treatment for urinary tract infection caused by ESBL-EC. The trial was registered with

  20. 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.

  1. A trial with IgY chicken antibodies to eradicate faecal carriage of Klebsiella pneumoniae and Escherichia coli producing extended-spectrum beta-lactamases

    Science.gov (United States)

    Jonsson, Anna-Karin; Larsson, Anders; Tängdén, Thomas; Melhus, Åsa; Lannergård, Anders

    2015-01-01

    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. PMID:26560861

  2. Efficacy of non-carbapenem antibiotics for pediatric patients with first febrile urinary tract infection due to extended-spectrum beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Abe, Yoshifusa; Inan-Erdogan, Işil; Fukuchi, Kunihiko; Wakabayashi, Hitomi; Ogawa, Yasuha; Hibino, Satoshi; Sakurai, Shunsuke; Matsuhashi, Kazuhiko; Watanabe, Yoshitaka; Hashimoto, Kaori; Ugajin, Kazuhisa; Itabashi, Kazuo

    2017-08-01

    Although carbapenem is the recommended for urinary tract infection (UTI) caused by extended-spectrum beta-lactamase (ESBL)-producing organisms, non-carbapenems have been reported to be effective for adult patients with UTI caused by ESBL-producing organisms. The purpose of this study was to evaluate the efficacy of non-carbapenems for pediatric patients with UTI due to ESBL-producing Escherichia coli (E. coli) based on the microbiologic and clinical outcomes. Fifteen children, who were treated for first febrile UTI caused by ESBL-producing E. coli were enrolled in this study. Antimicrobial susceptibilities and ESBL production were determined according to the Clinical and Laboratory Standards Institute guidelines. To detect CTX-M genes, polymerase chain reaction was performed with specific primers for bla CTX-M detection. Of the 15 enrolled patients, 10 (66.7%) were boys and 5 (33.3%) were girls, with a median age of four months. VUR was detected in six patients (40%). For detection of bla CTX-M by PCR, CTX-M-3, CTX-M-8, CTX-M-14, and CTX-M-15 were detected in five, one, eight, and one patient, respectively. Overall, 14 of the 15 isolates (93.3%) were susceptible for fosfomycin (FOM), and all isolates were susceptible for cefmetazole (CMZ), flomoxef (FMOX), and imipenem/cilastatin (IPM/CS). Of the 15 patients, 12 (80%) clinically improved without the use of carbapenems. In conclusion, even if isolates of ESBL-producing E. coli are multidrug resistant based on MIC assessment, clinical susceptibility to non-carbapenems, such as CMZ, FMOX, and FOM, is possible. Accordingly, carbapenems may not be required all the time for treatment of pediatric UTI in clinical practice. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  3. 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.

  4. Risk factors associated with the community-acquired colonization of extended-spectrum beta-lactamase (ESBL) positive Escherichia Coli. an exploratory case-control study.

    Science.gov (United States)

    Leistner, Rasmus; Meyer, Elisabeth; Gastmeier, Petra; Pfeifer, Yvonne; Eller, Christoph; Dem, Petra; Schwab, Frank

    2013-01-01

    The number of extended-spectrum beta-lactamase (ESBL) positive (+) Escherichia coli is increasing worldwide. In contrast with many other multidrug-resistant bacteria, it is suspected that they predominantly spread within the community. The objective of this study was to assess factors associated with community-acquired colonization of ESBL (+) E. coli. We performed a matched case-control study at the Charité University Hospital Berlin between May 2011 and January 2012. Cases were defined as patients colonized with community-acquired ESBL (+) E. coli identified language most commonly spoken at home (mother tongue). An additional rectal swab was obtained together with the questionnaire to verify colonization status. Genotypes of ESBL (+) E. coli strains were determined by PCR and sequencing. Risk factors associated with ESBL (+) E. coli colonization were analyzed by a multivariable conditional logistic regression analysis. We analyzed 85 cases and 170 controls, respectively. In the multivariable analysis, speaking an Asian language most commonly at home (OR = 13.4, CI 95% 3.3-53.8; p<0.001) and frequently eating pork (≥ 3 meals per week) showed to be independently associated with ESBL colonization (OR = 3.5, CI 95% 1.8-6.6; p<0.001). The most common ESBL genotypes were CTX-M-1 with 44% (n = 37), CTX-M-15 with 28% (n = 24) and CTX-M-14 with 13% (n = 11). An Asian mother tongue and frequently consuming certain types of meat like pork can be independently associated with the colonization of ESBL-positive bacteria. We found neither frequent consumption of poultry nor previous use of antibiotics to be associated with ESBL colonization.

  5. Collateral damage of flomoxef therapy: in vivo development of porin deficiency and acquisition of blaDHA-1 leading to ertapenem resistance in a clinical isolate of Klebsiella pneumoniae producing CTX-M-3 and SHV-5 beta-lactamases.

    Science.gov (United States)

    Lee, Chen-Hsiang; Chu, Chishih; Liu, Jien-Wei; Chen, Yi-Shung; Chiu, Chiung-Jung; Su, Lin-Hui

    2007-08-01

    The study aimed to characterize the genetic basis of flomoxef and collateral ertapenem resistance in a clinical isolate of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KP) after flomoxef exposure. Four ESBL-KP isolates (Lkp11-14) were recovered sequentially from four episodes of bacteraemia in an elderly patient. Laboratory investigations included genotyping by PFGE, resistance gene analysis by PCR and sequencing, and outer membrane protein analysis by SDS-PAGE. Plasmid analysis by DNA-DNA hybridization, electroporation and conjugation was also performed. Lkp14 was recovered after 21 days of flomoxef therapy. It demonstrated an indistinguishable PFGE pattern when compared with those produced by Lkp11-13. However, resistance to both flomoxef and ertapenem emerged in Lkp14. Molecular characterization revealed that, in addition to the pre-existing ESBL production (CTX-M-3 and SHV-5) and OmpK35 deficiency found in Lkp11-13, Lkp14 had acquired an extra plasmid-mediated AmpC beta-lactamase gene (blaDHA-1) and failed to express OmpK36, because of insertional inactivation by an insertion sequence IS5. Other resistance mechanisms, such as production of carbapenem-hydrolysing enzymes or expression of chromosomal efflux, were apparently not involved. Conjugational transfer of the plasmid-mediated blaDHA-1 gene into Lkp11 resulted in a significant increase in the MICs of cephamycins and beta-lactamase inhibitors, but not in those of carbapenems. Lkp14 was apparently derived from the previously flomoxef-susceptible isolates, Lkp11-13. After flomoxef exposure, the in vivo acquisition of the plasmid-mediated blaDHA-1 gene has led to flomoxef resistance in Lkp14, and the concomitant depletion of OmpK36 expression has resulted in a collateral effect of ertapenem resistance and diminished susceptibilities to imipenem and meropenem.

  6. Determinação da produção de metalo-beta-lactamases em amostras de Pseudomonas aeruginosa isoladas em João Pessoa, Paraíba

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    Santos Filho,Lauro; Santos,Isabele Beserra; Assis,Alexandro Mangueira Lima de; Xavier,Danilo Elias

    2002-01-01

    Bactérias produtoras de metalo-beta-lactamases (MBLs) são em grande parte resistentes aos betalactâmicos de largo espectro, incluindo oximino-aminotiazol cefalosporinas e também aos carbapenens. O objetivo deste trabalho foi detectar cepas de Pseudomonas aeruginosa resistentes ao imipenem e à ceftazidima, assim como identificar aquelas produtoras de MBLs. Foram estudadas 198 linhagens não-repetitivas isoladas de diversas amostras clínicas, hospitalares e comunitárias, identificadas bioquimica...

  7. The potential role of microbiota for controlling the spread of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE in neonatal population [version 1; referees: 2 approved

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    Thibaud Delerue

    2017-07-01

    Full Text Available The spread of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE in the hospital and also the community is worrisome. Neonates particularly are exposed to the risk of ESBL-PE acquisition and, owing to the immaturity of their immune system, to a higher secondary risk of ESBL-PE-related infection. Reducing the risk of acquisition in the hospital is usually based on a bundle of measures, including screening policies at admission, improving hand hygiene compliance, and decreasing antibiotic consumption. However, recent scientific data suggest new prevention opportunities based on microbiota modifications.

  8. 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

  9. Molecular epidemiology of outbreak-related pseudomonas aeruginosa strains carrying the novel variant blaVIM-17 metallo-beta-lactamase gene.

    Science.gov (United States)

    Siarkou, Victoria I; Vitti, Danai; Protonotariou, Efthimia; Ikonomidis, Alexandros; Sofianou, Danai

    2009-04-01

    A study was designed to investigate the molecular epidemiological characteristics of multidrug-resistant outbreak-related Pseudomonas aeruginosa isolates collected in a university hospital in northern Greece. Of 29 nonreplicate P. aeruginosa isolates resistant to carbapenems and ceftazidime, 14 were positive for metallo-beta-lactamase production. PCR analyses with primers specific for bla(VIM) and bla(IMP) revealed that 13 isolates carried a novel bla(VIM-2) gene variant, designated bla(VIM-17), and only 1 isolate carried bla(VIM-2), a gene predominant among P. aeruginosa strains in Greek hospitals. Pulsed-field gel electrophoresis of XbaI-digested genomic DNAs showed a close genetic relationship for 12 of 13 bla(VIM-17)-carrying outbreak-related isolates, which were of the O11 serotype; the clonally unrelated isolate carrying bla(VIM-17) was of the O12 serotype. PCR mapping strategies for the detection of class 1 integrons and sequencing approaches revealed the presence of integrons containing one bla(VIM) cassette flanked by two aacA29 cassettes. These integrons were similar but not identical to In59 (GenBank accession number AF263519) initially described in France. All isolates carrying bla(VIM-17), regardless of their genetic profile, had an identical integron, named In59.3, indicating that although the hospital outbreak was mainly due to clonal dissemination, the horizontal transmission of the bla(VIM-17)-containing integron among P. aeruginosa isolates should also have occurred. An outbreak-related isolate and a control strain, both of which carried the bla(VIM-2) gene but which were clonally distinct, had an identical integron, named In59.2, which differed only at the level of the bla(VIM) gene from In59.3 integrons, suggesting a common ancestry. The spread of the bla(VIM-17)-containing integron in clonally unrelated P. aeruginosa isolates without any evidence of plasmid carriage is probably associated with a transposon.

  10. Free water surface constructed wetlands limit the dissemination of extended-spectrum beta-lactamase producing Escherichia coli in the natural environment.

    Science.gov (United States)

    Vivant, Anne-Laure; Boutin, Catherine; Prost-Boucle, Stéphanie; Papias, Sandrine; Hartmann, Alain; Depret, Géraldine; Ziebal, Christine; Le Roux, Sophie; Pourcher, Anne-Marie

    2016-11-01

    The fates of Escherichia coli and extended-spectrum beta-lactamase-producing E. coli (ESBL E. coli) were studied over a period of one year in a free water surface constructed wetland (FWS CW) with a succession of open water zones and vegetation ponds (Typha or Phragmites), that received the effluent from a wastewater treatment plant. ESBL E. coli were detected and isolated from all sampling areas of the FWS CW throughout the study period. They represented 1‰ of the total E. coli population regardless of the origin of samples. Two main factors affected the log removal of E. coli and of ESBL E. coli: the season and the presence of vegetation. Between the inlet and the outlet of the FWS CW, the log removal of E. coli ranged from 1.5 in the warmer season (summer and fall) to 3.0 in the colder season (winter and spring). The concentrations of E. coli decreased significantly in the vegetated areas during the colder season, but increased in the warmer season, suggesting an effect of the plant growth stage on the survival of E. coli. Among the 369 ESBL E. coli isolates collected during our study, 84% harbored the CTX-M-ESBL type and 55.3% carried bla genes on plasmid DNA. Furthermore, 93% of the ESBL E. coli isolates were multidrug resistant but the proportion of resistant strains did not change significantly along the FWS CW. ESBL E. coli were characterized by MLST analysis using the 7 genes based Achtman Scheme. ESBL E. coli isolated from water, sediments, roots and feces of myocastors collected in the FWS CW and in the recipient river were genotypically related, suggesting persistence and circulation of the ESBL producing E. coli throughout the FWS CW and in the receiving river. Overall, these observations show that FWS CW could be an efficient treatment for ESBL E. coli disinfection of wastewater and could limit their dissemination in the aquatic environment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. 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

  12. Imported chicken meat as a potential source of quinolone-resistant Escherichia coli producing extended-spectrum beta-lactamases in the UK.

    Science.gov (United States)

    Warren, R E; Ensor, V M; O'Neill, P; Butler, V; Taylor, J; Nye, K; Harvey, M; Livermore, D M; Woodford, N; Hawkey, P M

    2008-03-01

    Escherichia coli producing CTX-M-15 enzyme began to rapidly spread in the UK from around 2003 but other types also occur, notably CTX-M-14. We examined breasts from UK-reared (n = 62) and imported (n = 27) chickens as potential sources of quinolone-resistant E. coli with bla(CTX-M) genes. A further 40 samples for which the country of rearing could not be identified were examined. During 2006, 129 fresh and frozen chicken breast fillets were purchased from retail outlets in the West Midlands. These were cultured for E. coli on CLED agar containing 8 mg/L ciprofloxacin and carrying a 10 microg cefpodoxime disc. Resistant isolates were identified and typed by RAPD fingerprinting; bla(CTX-M) was identified by PCR and genotyped by reverse-line hybridization. The country of rearing was identified from the packaging for 89 of 129 purchased samples. Only one of the 62 UK-reared chicken samples carried E. coli producing a CTX-M-1 enzyme, whereas 10 of 27 samples reared overseas had E. coli with CTX-M enzymes. Specifically, 4/10 Brazilian, 3/4 Brazilian/Polish/French, and 2/2 Dutch samples had E. coli with CTX-M-2 enzymes. Six of 40 samples for which the country of rearing was not known had producers of CTX-M enzymes, 5 of them with CTX-M-14. Quinolone-resistant E. coli with various CTX-M beta-lactamase genes that are common in human infections worldwide were found in imported chicken breasts, indicating a possible source for gut colonization. Samples from Brazil were commonly positive for E. coli with CTX-M-2, the dominant bla(CTX-M) genotype from human infections in South America, which is currently rare in clinical infections in the UK. CTX-M-15, the dominant CTX-M type in human infections in the UK, was not found in chicken isolates, suggesting that the UK-reared chickens are not a reservoir of CTX-M-15.

  13. Risk factors associated with the community-acquired colonization of extended-spectrum beta-lactamase (ESBL positive Escherichia Coli. an exploratory case-control study.

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    Rasmus Leistner

    Full Text Available BACKGROUND: The number of extended-spectrum beta-lactamase (ESBL positive (+ Escherichia coli is increasing worldwide. In contrast with many other multidrug-resistant bacteria, it is suspected that they predominantly spread within the community. The objective of this study was to assess factors associated with community-acquired colonization of ESBL (+ E. coli. METHODS: We performed a matched case-control study at the Charité University Hospital Berlin between May 2011 and January 2012. Cases were defined as patients colonized with community-acquired ESBL (+ E. coli identified <72 h after hospital admission. Controls were patients that carried no ESBL-positive bacteria but an ESBL-negative E.coli identified <72 h after hospital admission. Two controls per case were chosen from potential controls according to admission date. Case and control patients completed a questionnaire assessing nutritional habits, travel habits, household situation and language most commonly spoken at home (mother tongue. An additional rectal swab was obtained together with the questionnaire to verify colonization status. Genotypes of ESBL (+ E. coli strains were determined by PCR and sequencing. Risk factors associated with ESBL (+ E. coli colonization were analyzed by a multivariable conditional logistic regression analysis. RESULTS: We analyzed 85 cases and 170 controls, respectively. In the multivariable analysis, speaking an Asian language most commonly at home (OR = 13.4, CI 95% 3.3-53.8; p<0.001 and frequently eating pork (≥ 3 meals per week showed to be independently associated with ESBL colonization (OR = 3.5, CI 95% 1.8-6.6; p<0.001. The most common ESBL genotypes were CTX-M-1 with 44% (n = 37, CTX-M-15 with 28% (n = 24 and CTX-M-14 with 13% (n = 11. CONCLUSION: An Asian mother tongue and frequently consuming certain types of meat like pork can be independently associated with the colonization of ESBL-positive bacteria. We found neither frequent consumption

  14. Extended-Spectrum-Beta-Lactamase- and Plasmid-Encoded Cephamycinase-Producing Enterobacteria in the Broiler Hatchery as a Potential Mode of Pseudo-Vertical Transmission.

    Science.gov (United States)

    Projahn, Michaela; Daehre, Katrin; Roesler, Uwe; Friese, Anika

    2017-01-01

    Antimicrobial resistance through extended-spectrum beta-lactamases (ESBLs) and transferable (plasmid-encoded) cephamycinases (pAmpCs) represents an increasing problem in human and veterinary medicine. The presence of ESBL-/pAmpC-producing commensal enterobacteria in farm animals, such as broiler chickens, is considered one possible source of food contamination and could therefore also be relevant for human colonization. Studies on transmission routes along the broiler production chain showed that 1-day-old hatchlings are already affected. In this study, ESBL-/pAmpC-positive broiler parent flocks and their corresponding eggs, as well as various environmental and air samples from the hatchery, were analyzed. The eggs were investigated concerning ESBL-/pAmpC-producing enterobacteria on the outer eggshell surface (before/after disinfection), the inner eggshell surface, and the egg content. Isolates were analyzed concerning their species, their phylogroup in the case of Escherichia coli strains, the respective resistance genes, and the phenotypical antibiotic resistance. Of the tested eggs, 0.9% (n = 560) were contaminated on their outer shell surface. Further analyses using pulsed-field gel electrophoresis showed a relationship of these strains to those isolated from the corresponding parent flocks, which demonstrates a pseudo-vertical transfer of ESBL-/pAmpC-producing enterobacteria into the hatchery. Resistant enterobacteria were also found in environmental samples from the hatchery, such as dust or surfaces which could pose as a possible contamination source for the hatchlings. All 1-day-old chicks tested negative directly after hatching. The results show a possible entry of ESBL-/pAmpC-producing enterobacteria from the parent flocks into the hatchery; however, the impact of the hatchery on colonization of the hatchlings seems to be low. ESBL-/pAmpC-producing enterobacteria occur frequently in broiler-fattening farms. Recent studies investigated the prevalence and

  15. First report on extended-spectrum beta-lactamase (ESBL) producing Escherichia coli from European free-tailed bats (Tadarida teniotis) in Portugal: A one-health approach of a hidden contamination problem.

    Science.gov (United States)

    Garcês, Andreia; Correia, Susana; Amorim, Francisco; Pereira, José Eduardo; Igrejas, Gilberto; Poeta, Patrícia

    2017-12-23

    The main aim of this study was to characterize the diversity of extended-spectrum beta-lactamases (ESBLs) in Escherichia coli isolates from European free tailed-bats (Tadarida teniotis) in Portugal. ESBL-producing E. coli isolates were recovered from 14 of 146 faecal samples (9.6%) and a total of 19 isolates were completely characterized. The more prevalent beta-lactamase genes detected were bla CTX-M-1 (57.9%) and bla CTX-M-3 (36.8%), followed by bla SHV (31.6%), bla TEM (21.1%), bla OXA (10.5%) and bla CTX-M-9 (10.5%). Among other associated resistance genes studied, tet(A) and tet(B) were predominant and fimA was the main virulence factor detected. Phylogroups D (47.4%) and A (31.6%) were the more prevalent, followed by group B2 (21.1%). Bats are reservoirs of antimicrobial-resistant bacteria and resistance determinants and is important in further studies to identify the main sources of pollution in the environment, such as water or insects that may contain these multiresistant organisms. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. 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.

    Directory of Open Access Journals (Sweden)

    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.

  17. 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

    Directory of Open Access Journals (Sweden)

    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.

  18. 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

    Directory of Open Access Journals (Sweden)

    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

  19. 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

    Science.gov (United States)

    Schauss, Thorsten; Glaeser, Stefanie P.; Gütschow, Alexandra; Dott, Wolfgang; Kämpfer, Peter

    2015-01-01

    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 bla CTX-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. coli in

  20. 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

  1. 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

    1996-01-01

    BACKGROUND: Antibodies against chromosomal beta-lactamase of Pseudomonas aeruginosa (a beta ab) are markers of the development of resistance of P aeruginosa to beta-lactam antibiotics in patients with cystic fibrosis and chronic lung infection. The role of these antibodies in patients with chronic...... 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...... complexes might be less important than other antipseudomonal antibodies. A beneficial neutralising effect of the a beta ab antibodies on the antibiotic destroying enzymes may be an additional factor....

  2. Distribution and Antimicrobial Susceptibility Pattern of Gram Negative Bacteria Causing Urinary Tract Infection (UTI and Detection New Delhi Metallo-beta-lactamase-1 (NDM-1 Producing Isolates in Ahwaz

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    Parviz Afrugh

    2016-04-01

    Full Text Available Background: Urinary tract infection (UTI is the commonest bacterial infectious disease in worldwide (especially in developing countries with a high rate of morbidity and financial cost. The management of UTI infections has been jeopardized by increase in immergence of antimicrobial drug resistance. Knowledge of the local bacterial etiology and susceptibility patterns is required to trace any change that might have occurred in time so that updated recommendation for optimal empirical therapy of UTI can be made. The aim of this investigation was distribution and antimicrobial susceptibility pattern of gram negative bacteria causing urinary tract infection (UTI and detection NDM-1 (new-delhi-metallo-beta-lactamase-1 producing isolates in Ahwaz. Materials and Methods: This cross-sectional study was done during a period of one year from April 2013 to March 2014. Clean catch midstream urine samples were collected from suspected patients to UTI. The isolates were identified based on morphological and biochemical testes. Culture was performed on routine microbiological media. Susceptibility testing was performed according CLSI (2013 guidelines. Detection of carbapenemase producing isolates was performed by modified hodge test (MHT. Metallo-beta-lactamase isolates were detected by imipenem-EDTA combined disc test (CDT. Results: In this study 708 gram negative organisms were isolated from urine samples. E.coli was the most common isolated bacteria (67% followed by Klebsiella spp. (26.5% and Enterobacter spp. (2.5%. In antibiotic susceptibility testing more than 90% of isolates were sensitive to tetracycline, ceftazidime, meropenem, amikacin, cefotaxime, imipenem, and cefepime. Isolates were more resistant to cephalothin (32%, co-trimoxazol (30.5%, and nalidixic acid (25%. Conclusion: In our results isolated organisms from outpatients showed very high sensitivity to common antibiotics. Continuous and regular monitoring of susceptibility pattern of

  3. Horizontol dissemination of TEM- and SHV-typr beta-lactamase genes-carrying resistance plasmids amongst clonical isolates of Enterobacteriaceae Disseminação horizontal de plasmídios de resistência contendo genes de beta-lactamase dos tipos TEM e SHV entre isolados clínicos de Enterobacteriaceae

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    Osman Birol Ozgumus

    2008-12-01

    Full Text Available The extended-spectrum β-lactamase (ESBL-producing bacteria have been isolated at increasing frequency worldwide. Expression of ESBL is often associated with multidrug resistance and dissemination by resistance plasmids. During a two-month period in 2000, 133 clinical isolates of enterobacterial strains were randomly collected from outpatients and inpatients at a university hospital in Turkey. The ESBL producing strains were determined by double-disk synergy (DDS testing. Twenty ESBL producing strains (15% including Escherichia coli (n = 9, Klebsiella pneumoniae (n = 7, Klebsiella oxytoca (n = 2 and Enterobacter aerogenes (n = 2 were detected and further analyzed for their resistance transfer features, plasmid profile and nature of the resistance genes. Plasmid transfer assays were performed using broth mating techniques. TEM- and SHV- genes were analyzed by polymerase chain reaction (PCR and hybridization using specific probes. EcoRI restriction enzyme analyses of R plasmids were used in the detection of epidemic plasmids. Fourteen plasmid profiles (A, B1, B2, C1, and C2 to L were obtained with EcoRI restriction enzyme analysis. Most of these plasmids were detected to carry both TEM- and SHV-derived genes by PCR, and confirmed by localizing each gene by hybridization assay. Epidemiological evidence indicated that there was an apparent horizontal dissemination of conjugative R plasmids among multidrug-resistant enterobacterial genera and species in this hospitalO isolamento de bactérias produtoras de beta-lactamases de espectro expandido (ESBL está aumentando no mundo todo. Freqüentemente, a expressão de ESBL está associada com resistência a múltiplas drogas e disseminação por plasmídios de resistência. Durante um período de dois meses em 2000, 133 isolados clínicos de cepas de enterobactérias foram obtidos aleatoriamente de pacientes internos e externos de um hospital universitário na Turquia. As cepas produtoras de ESBL foram

  4. 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

  5. Complete Sequence of p07-406, a 24,179-base-pair plasmid harboring the blaVIM-7 metallo-beta-lactamase gene in a Pseudomonas aeruginosa isolate from the United States.

    Science.gov (United States)

    Li, Hongyang; Toleman, Mark A; Bennett, Peter M; Jones, Ronald N; Walsh, Timothy R

    2008-09-01

    An outbreak involving a Pseudomonas aeruginosa strain that was resistant to all tested antimicrobials except polymyxin B occurred in a hospital in Houston, TX. Previous studies on this strain showed that it possesses a novel mobile metallo-beta-lactamase (MBL) gene, designated bla(VIM-7), located on a plasmid (p07-406). Here, we report the complete sequence, annotation, and functional characterization of this plasmid. p07-406 is 24,179 bp in length, and 29 open reading frames were identified related to known or putatively recognized proteins. Analysis of this plasmid showed it to be comprised of four distinct regions: (i) a region of 5,200 bp having a Tn501-like mercuric resistance (mer) transposon upstream of the replication region; (ii) a Tn3-like transposon carrying a truncated integron with a bla(VIM-7) gene and an insertion sequence inserted at the other end of this transposon; (iii) a region of four genes, upstream of the Tn3-like transposon, possessing very high similarity to plasmid pXcB from Xanthomonas campestris pv. citri commonly associated with plants; (iv) a backbone sequence similar to the backbone structure of the IncP group plasmid Rms149, pB10, and R751. This is the first plasmid to be sequenced carrying an MBL gene and highlights the amelioration of DNA segments from disparate origins, most noticeably from plant pathogens.

  6. Foreign travel is a major risk factor for colonization with Escherichia coli producing CTX-M-type extended-spectrum beta-lactamases: a prospective study with Swedish volunteers.

    Science.gov (United States)

    Tängdén, Thomas; Cars, Otto; Melhus, Asa; Löwdin, Elisabeth

    2010-09-01

    Foreign travel has been suggested to be a risk factor for the acquisition of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae. To our knowledge, this has not previously been demonstrated in a prospective study. Healthy volunteers traveling outside Northern Europe were enrolled. Rectal swabs and data on potential travel-associated risk factors were collected before and after traveling. A total of 105 volunteers were enrolled. Four of them did not complete the study, and one participant carried ESBL-producing Escherichia coli before travel. Twenty-four of 100 participants with negative pretravel samples were colonized with ESBL-producing Escherichia coli after the trip. All strains produced CTX-M enzymes, mostly CTX-M-15, and some coproduced TEM or SHV enzymes. Coresistance to several antibiotic subclasses was common. Travel to India was associated with the highest risk for the acquisition of ESBLs (88%; n = 7). Gastroenteritis during the trip was an additional risk factor (P = 0.003). Five of 21 volunteers who completed the follow-up after 6 months had persistent colonization with ESBLs. This is the first prospective study demonstrating that international travel is a major risk factor for colonization with ESBL-producing Enterobacteriaceae. Considering the high acquisition rate of 24%, it is obvious that global efforts are needed to meet the emergence and spread of CTX-M enzymes and other antimicrobial resistances.

  7. High-resolution melting-curve analysis of ligation-mediated real-time PCR for rapid evaluation of an epidemiological outbreak of extended-spectrum-beta-lactamase-producing Escherichia coli.

    Science.gov (United States)

    Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E; Schön, Thomas

    2011-12-01

    Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE.

  8. High-Resolution Melting-Curve Analysis of Ligation-Mediated Real-Time PCR for Rapid Evaluation of an Epidemiological Outbreak of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli ▿

    Science.gov (United States)

    Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E.; Schön, Thomas

    2011-01-01

    Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE. PMID:21956981

  9. Should the patients colonized with extended-spectrum beta-lactamase-producing Gram-negative bacilli (E-GNB) coming to hospital from the community with pneumonia get anti-E-GNB active empirical treatment?

    Science.gov (United States)

    Peterlin, Lara; Žagar, Mateja; Lejko Zupanc, Tatjana; Paladin, Marija; Beović, Bojana

    2017-10-01

    Extended-spectrum beta-lactamases are responsible for resistance of Gram-negative bacilli to several beta-lactam antibiotics, including those prescribed for treatment pneumonia. To evaluate the importance of colonization with E-GNB for the choice of empirical treatment we performed a retrospective case-control study including 156 patients, hospitalized for treatment of pneumonia from 2009 through 2013. Empirical treatment success and in-hospital survival were significantly lower in patients colonized with E-GNB compared to non-colonized (p = 0.002, p = 0.035). When comparing subgroups of colonized patients, treatment success was significantly lower in patients who were colonized with E-GNB resistant to empirical antibiotic (p = 0.010), but not in those colonized by E-GNB susceptible to empirically given antibiotic (p = 0.104). Difference in in-hospital mortality was insignificant in both subgroups (p = 0.056, p = 0.331). The results of study suggest that an anti-E-GNB active antibiotic should be used for empirical treatment of pneumonia in E-GNB colonized patients.

  10. Environmental pollution with antimicrobial agents from bulk drug manufacturing industries in Hyderabad, South India, is associated with dissemination of extended-spectrum beta-lactamase and carbapenemase-producing pathogens.

    Science.gov (United States)

    Lübbert, Christoph; Baars, Christian; Dayakar, Anil; Lippmann, Norman; Rodloff, Arne C; Kinzig, Martina; Sörgel, Fritz

    2017-08-01

    High antibiotic and antifungal concentrations in wastewater from anti-infective drug production may exert selection pressure for multidrug-resistant (MDR) pathogens. We investigated the environmental presence of active pharmaceutical ingredients and their association with MDR Gram-negative bacteria in Hyderabad, South India, a major production area for the global bulk drug market. From Nov 19 to 28, 2016, water samples were collected from the direct environment of bulk drug manufacturing facilities, the vicinity of two sewage treatment plants, the Musi River, and habitats in Hyderabad and nearby villages. Samples were analyzed for 25 anti-infective pharmaceuticals with liquid chromatography-tandem mass spectrometry and for MDR Gram-negative bacteria using chromogenic culture media. In addition, specimens were screened with PCR for bla VIM , bla KPC , bla NDM , bla IMP-1 , and bla OXA-48 resistance genes. All environmental specimens from 28 different sampling sites were contaminated with antimicrobials. High concentrations of moxifloxacin, voriconazole, and fluconazole (up to 694.1, 2500, and 236,950 µg/L, respectively) as well as increased concentrations of eight other antibiotics were found in sewers in the Patancheru-Bollaram industrial area. Corresponding microbiological analyses revealed an extensive presence of extended-spectrum beta-lactamase and carbapenemase-producing Enterobacteriaceae and non-fermenters (carrying mainly bla OXA-48 , bla NDM , and bla KPC ) in more than 95% of the samples. Insufficient wastewater management by bulk drug manufacturing facilities leads to unprecedented contamination of water resources with antimicrobial pharmaceuticals, which seems to be associated with the selection and dissemination of carbapenemase-producing pathogens. The development and global spread of antimicrobial resistance present a major challenge for pharmaceutical producers and regulatory agencies.

  11. Characterization of Silver Nanomaterials Derived from Marine Streptomyces sp. Al-Dhabi-87 and Its In Vitro Application against Multidrug Resistant and Extended-Spectrum Beta-Lactamase Clinical Pathogens

    Directory of Open Access Journals (Sweden)

    Naif Abdullah Al-Dhabi

    2018-04-01

    Full Text Available A novel antagonistic marine Streptomyces sp. Al-Dhabi-87 that was recovered from the Gulf region of Saudi Arabia was used to synthesize silver nanoparticles (NP from the culture free extract. The produced NP were confirmed by UV-visible spectroscopy (UV-Vis, high-resolution scanning electron microscope (HRSEM, transmission electron microscope (TEM, Fourier-transform infrared spectroscopy (FTIR, Energy Dispersive Spectroscopy (EDAX, and X-ray Powder Diffraction (XRD, and broth micro dilution techniques were employed for the determination of minimum inhibitory concentrations (MIC values. The synthesized NP was authenticated by alterations in color and wavelength scanning. HRSEM and TEM analysis confirmed that the size of the NP ranged from 10 to 17 nm and that it was spherical in shape. In addition, the FTIR spectrum revealed a variation in the band values from 500 to 3300 cm−1 respectively. Rietveld refinement analysis of the XRD data confirmed the size of the NP, which coincided with the results of the TEM analysis. In addition, the Riveted refinement analysis supported the TEM data. The NP documented significant activity against the wound infection microbial strains, such as Enterococcus faecalis, Staphylococcus epidermidis, and Staphylococcus aureus. Gram negative bacteria, such as Pseudomonas aeruginosa, Klebsiella pneumonia, and Escherichia coli revealed MIC values of 0.039, 0.078, and 0.152 mg/mL, respectively. The promising activity of NP towards extended-spectrum beta-lactamases E.coli, drug resistant Acinetobacter baumannii, and multidrug resistant S. aureus (at 0.018, 0.039, and 0.039 mg/mL, respectively was advantageous. Overall, NP that were obtained from the novel Streptomyces sp. Al-Dhabi-87, with its promising antimicrobial activity towards the drug resistant pathogens, would be useful for healing infectious diseases.

  12. Discrepancy between effects of carbapenems and flomoxef in treating nosocomial hemodialysis access-related bacteremia secondary to extended spectrum beta-lactamase producing Klebsiella pneumoniae in patients on maintenance hemodialysis.

    Science.gov (United States)

    Yang, Chih-Chao; Li, Shau-Hsuan; Chuang, Feng-Rong; Chen, Chih-Hung; Lee, Chih-Hsiung; Chen, Jin-Bor; Wu, Chien-Hsing; Lee, Chien-Te

    2012-09-05

    Hemodialysis (HD) patients are susceptible to extended spectrum beta-lactamase (ESBL)-producing bacterial infections. Because the optimal treatment and clinical significance of ESBL-producing Klebsiella pneumoniae (ESBL-Kp) HD access-related bacteremia remain unclear, we conducted this retrospective study to determine the clinical outcomes of patients treated with either flomoxef or a carbapenem. The eligibility criterion was fistula or graft- or catheter- related ESBL-Kp bacteremia in patients on maintenance HD. The clinical characteristics and antibiotic management were analyzed. Outcome was determined by mortality resulting from bacteremia during the 14-day period after the first positive blood culture for flomoxef-susceptible ESBL-Kp. The 57 patients studied were predominantly elderly, malnourished, with a history of severe illnesses and broad-spectrum antibiotic use before the onset of bacteremia, and with severe septicemia as determined by the Pitt bacteremia score (PBS). The study population comprised 7 fistula, 8 graft, and 42 HD catheter-related bacteremia (CRB) cases, and the mortality rate was high (36/57, 63.2%) in these 57 patients. Of 42 patients with CRB, those in the deceased group (27/42, 64.3%) had significantly lower levels of serum albumin, longer prior hospital stay and duration of catheter-dependent HD, and higher PBS than patients in the survived group. Failure to receive effective antibiotics (flomoxef or a carbapenem) within 5 days after onset of bacteremia and treatment with flomoxef both significantly contributed to higher mortality. Multivariate analyses revealed that flomoxef use, PBS, and catheter-dependent HD >30 days were independently associated with increased mortality (OR, 3.52; 95% CI, 1.19-58.17, OR, 2.92; 95% CI, 1.36-6.26 and OR, 5.73; 95% CI, 1.21-63.2, respectively). Considering the high mortality rate, ESBL-Kp should be recognized as a possible pathogen in patients on maintenance HD at high risk of acquiring HD access

  13. Molecular epidemiology of Escherichia coli producing extended-spectrum {beta}-lactamases in Lugo (Spain): dissemination of clone O25b:H4-ST131 producing CTX-M-15.

    Science.gov (United States)

    Blanco, Miguel; Alonso, Maria Pilar; Nicolas-Chanoine, Marie-Hélène; Dahbi, Ghizlane; Mora, Azucena; Blanco, Jesús E; López, Cecilia; Cortés, Pilar; Llagostera, Montserrat; Leflon-Guibout, Véronique; Puentes, Beatriz; Mamani, Rosalía; Herrera, Alexandra; Coira, María Amparo; García-Garrote, Fernando; Pita, Julia María; Blanco, Jorge

    2009-06-01

    Having shown that the Xeral-Calde Hospital in Lugo (Spain) has been concerned by Escherichia coli clone O25:H4-ST131 producing CTX-M-15 (Nicolas-Chanoine et al. J Antimicrob Chemother 2008; 61: 273-81), the present study was carried out to evaluate the prevalence of this clone among the extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates and also to molecularly characterize the E. coli isolates producing ESBL other than CTX-M-15. In the first part of this study, 105 ESBL-producing E. coli isolates (February 2006 to March 2007) were characterized with regard to ESBL enzymes, serotypes, virulence genes, phylogenetic groups, multilocus sequence typing (MLST) and PFGE. In the second part of this study, 249 ESBL-producing E. coli isolates (April 2007 to May 2008) were investigated only for the detection of clone O25b:H4-ST131 producing CTX-M-15 using a triplex PCR developed in this study and based on the detection of the new operon afa FM955459 and the targets rfbO25b and 3' end of the bla(CTX-M-15) gene. Of the 105 ESBL-producing E. coli isolates, 60 (57.1%) were positive for CTX-M-14, 23 (21.9%) for CTX-M-15, 10 (9.5%) for SHV-12 and 7 (6.7%) for CTX-M-32. Serotypes, virulence genes, phylogenetic groups and molecular typing by PFGE demonstrated high homogeneity within those producing CTX-M-15 and high diversity within E. coli producing CTX-M-14 and other ESBLs. By PFGE, CTX-M-15-producing E. coli isolates O25b:H4 belonging to the phylogenetic group B2 and MLST profile ST131 were grouped in the same cluster. The epidemic strain of clone O25b:H4-ST131 represented 23.1%, 22.5% and 20.0% of all ESBL-producing E. coli isolated in 2006, 2007 and 2008, respectively. CTX-M-type ESBLs, primarily CTX-M-14 and CTX-M-15, have emerged as the predominant types of ESBL produced by E. coli isolates in Lugo. In view of the reported findings, long-term care facilities for elderly people may represent a significant reservoir for E. coli clone O25b:H4-ST131 producing CTX

  14. 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

    Directory of Open Access Journals (Sweden)

    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.

  15. Discrepancy between effects of carbapenems and flomoxef in treating nosocomial hemodialysis access-related bacteremia secondary to extended spectrum beta-lactamase producing klebsiella pneumoniae in patients on maintenance hemodialysis

    Directory of Open Access Journals (Sweden)

    Yang Chih-Chao

    2012-09-01

    Full Text Available Abstract Background Hemodialysis (HD patients are susceptible to extended spectrum beta-lactamase (ESBL-producing bacterial infections. Because the optimal treatment and clinical significance of ESBL-producing Klebsiella pneumoniae (ESBL-Kp HD access-related bacteremia remain unclear, we conducted this retrospective study to determine the clinical outcomes of patients treated with either flomoxef or a carbapenem. Methods The eligibility criterion was fistula or graft- or catheter- related ESBL-Kp bacteremia in patients on maintenance HD. The clinical characteristics and antibiotic management were analyzed. Outcome was determined by mortality resulting from bacteremia during the 14‐day period after the first positive blood culture for flomoxef-susceptible ESBL-Kp. Results The 57 patients studied were predominantly elderly, malnourished, with a history of severe illnesses and broad-spectrum antibiotic use before the onset of bacteremia, and with severe septicemia as determined by the Pitt bacteremia score (PBS. The study population comprised 7 fistula, 8 graft, and 42 HD catheter-related bacteremia (CRB cases, and the mortality rate was high (36/57, 63.2% in these 57 patients. Of 42 patients with CRB, those in the deceased group (27/42, 64.3% had significantly lower levels of serum albumin, longer prior hospital stay and duration of catheter-dependent HD, and higher PBS than patients in the survived group. Failure to receive effective antibiotics (flomoxef or a carbapenem within 5 days after onset of bacteremia and treatment with flomoxef both significantly contributed to higher mortality. Multivariate analyses revealed that flomoxef use, PBS, and catheter-dependent HD >30 days were independently associated with increased mortality (OR, 3.52; 95% CI, 1.19–58.17, OR, 2.92; 95% CI, 1.36–6.26 and OR, 5.73; 95% CI, 1.21–63.2, respectively. Conclusions Considering the high mortality rate, ESBL-Kp should be recognized as a possible pathogen in

  16. 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

    Directory of Open Access Journals (Sweden)

    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

  17. Clinical patterns, epidemiology and risk factors of community-acquired urinary tract infection caused by extended-spectrum beta-lactamase producers: a prospective hospital case-control study.

    Science.gov (United States)

    Almomani, Basima A; Hayajneh, Wail A; Ayoub, Abeer M; Ababneh, Mera A; Al Momani, Miral A

    2018-05-10

    To assess incidence rate, risk factors and susceptibility patterns associated with extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli or Klebsiella pneumoniae in community-acquired urinary tract infections (CA-UTIs). A prospective, case-control study was conducted at a tertiary teaching hospital from Jan 2015 to Dec 2016. The results of microbiology cultures were initially screened to include only patients with positive E. coli or K. pneumoniae urine cultures. Afterwards, clinical symptoms were assessed to confirm the UTI. To investigate the risk factors, patients with a positive urine culture for ESBL-producing isolates were assigned as cases, while patients with non-ESBL were assigned as controls. Out of 591 patients included in this study, 57.5% (n = 340) were included in the control group and 42.5% (n = 251) were in the case group. The incidence rate of ESBL-producing isolates was 3.465 cases per 1000-patient hospital admissions. Male gender (OR = 1.856, 95% CI = 1.192-2.889, p = 0.006), pediatrics (OR = 1.676, 95% CI = 1.117-2.517, p = 0.013), patients with comorbidity (OR = 1.542, 95% CI = 1.029-2.312, p = 0.036) and UTI in the previous 12 months (OR = 1.705, 95% CI = 1.106-2.628, p = 0.016) were independently associated with a higher risk of infection. The resistance rate for most commonly prescribed antibiotics was high. Our results suggest that the incidence of ESBL producers among CA-UTIs is high. Male gender, pediatrics, comorbidity and UTI in the previous 12 months were associated with a higher risk for infection. Continuous surveillance and prudent antibiotic use by healthcare professionals are important factors for effective control of ESBL associated infections.

  18. Elaboration and evaluation of a new screening medium for detection and presumptive identification of extended-spectrum beta-lactamase-producing organisms (ESBL Elaboração e avaliação de um novo meio de triagem para a detecção e identificação presuntiva de enterobactérias produtoras de beta-lactamase de espectro estendido (ESBL

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    Carlos Henrique Pessôa de Menezes e Silva

    2000-10-01

    Full Text Available The new ß-lactamases have arisen largely as a consequence of heavy use of new expanded spectrum ß-lactam antibiotics. Health professionals need to be aware of the resistance problems caused by the new enzymes, and know the correct procedures to detect, prevent and control such problems. In this study, a new screening medium called Ceftazidime-Inositol-Vancomycin-Amphotericin B Agar (CIVA was elaborated and the microbiological performance was evaluated for the detection and presumptive identification of ESBL-producing members of Enterobacteriaceae. It was performed in 126 stool samples from hospitalized patients at Santa Monica Hospital (Vila Velha, ES, Brazil, who had been heavily exposed to broad-spectrum antibiotic combinations. The bacteria were detected by the medium based on their colony colours (due to inositol fermentation. Additional tests were required for correct identification of these strains. No false positive rates were detected.As novas beta-lactamases têm aparecido, na maioria das vezes, como consequência do amplo uso de antibióticos beta-lactâmicos de largo espectro. Os profissionais de saúde precisam estar atentos acerca do problema da resistência bacteriana causado por estas novas enzimas e conhecer os corretos procedimentos para detectar, prevenir e controlar tais situações. Neste estudo, um novo meio de triagem foi desenvolvido no setor de Microbiologia do Marcos Daniel Laboratório - Vitória, ES (denominado CIVA - Ceftazidima-Inositol-Vancomicina-Anfotericina B e a sua performance microbiológica foi avaliada quanto à detecção e identificação presuntiva de enterobactérias produtoras de ESBL. Foram realizadas 126 culturas de amostras fecais de pacientes hospitalizados no Hospital Santa Monica (Vila Velha, ES, Brasil, previamente expostos a combinações de antibióticos de largo espectro. As bactérias foram detectadas pelo meio baseado nas cores das colônias desenvolvidas (devido à fermentação do

  19. Split Beta-Lactamase Complementation Assay

    Indian Academy of Sciences (India)

    IAS Admin

    A Search for the Molecular Better Half! Vaishali Verma ... These assays comprise of a protein molecule, ... ciferase, beta-galactosidase, GFP, g3p of M13 filamentous ph- .... sensors of protein–protein interactions, Nature Biotechnology, Vol.20,.

  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

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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

  1. Detection of metallo-beta-lactamase producing Pseudomonas ...

    African Journals Online (AJOL)

    sunny t

    2016-02-24

    Feb 24, 2016 ... A big concern with using these methods is the direct bactericidal effect of EDTA on .... comparison; it was about 20 mm (data not shown). The diminution ... disadvantages of the DDST is the subjective interpretation of results ...

  2. Extended spectrum beta-lactamase producing Uropathogens in ...

    African Journals Online (AJOL)

    ESBLs contribute to multi drug resistance among the organisms and the detection of ESBLs is ... the antibiogram pattern of ESBLs producing isolates to enable better treatment. ... Secondary Health facility providing antenatal care for pregnant women. ... Key Words: ESBLs, asymptomatic bacteriuria and multidrug resistance.

  3. Distribution Profile of Extended Spectrum Beta Lactamase (ESBL ...

    African Journals Online (AJOL)

    ADOWIE PERE

    organism among Enterobacteriaceae in human diseases and causes serious and common infections such as .... outbreaks but is also mostly found in community- acquired .... of community- acquired pneumonia amongst adults in the Asia-.

  4. Resistance Pattern and Detection of Metallo-beta-lactamase Genes ...

    African Journals Online (AJOL)

    2018-02-23

    Feb 23, 2018 ... Background: Acquired metallo-β-lactamases (MBLs) pose serious problem both in terms of ... P. aeruginosa from clinical samples submitted to the Medical Microbiology ... pan-drug-resistant .... Phenotypically confirmed MBL producers were stored ... 103 (51.5%); ear swab 32 (16%); urine 27 (13.5%); and.

  5. Phenotypic Detection of extended spectrum beta lactamase and ...

    African Journals Online (AJOL)

    Conclusion: The finding of the study therefore indicates that carbapenem resistance is mediated by carbapenemase production and or overproduction of ESBL coupled with reduced porins. Co-production of carbapenemase, MBLs and ESBLs by some of the isolates is worrisome. Susceptibility to colistin and tigecycline was ...

  6. New Delhi Metallo – beta lactamase – 1 containing ...

    African Journals Online (AJOL)

    One of the biggest problems associated with the antibiotic therapy is resistance. Recently published studies have revealed that enterobacteriaceae, like E. coli and Klebsiella, isolated from several Indian centers are resistant to many antibiotics including some highly potent antibiotics like carbapenems. It has been proposed ...

  7. Detection of metallo-beta-lactamase producing Pseudomonas ...

    African Journals Online (AJOL)

    sunny t

    2016-02-24

    Feb 24, 2016 ... Since the increasing prevalence of carbapenem-resistant Pseudomonas aeruginosa spp., accurate detection of metallo-β-lactamase (MBL) such as blaVIM type enzyme producing isolates became very important. However, phenotypic MBL detection methods previously reported are not highly sensitive or.

  8. Detection of Amp C Beta Lactamases in Clinical Isolates of ...

    African Journals Online (AJOL)

    A total of 81 consecutive non repetitive clinical isolates of Escherichia coli (n=40) and Klebsiella spp. (n=41) were screened for AmpC production by disc diffusion method using cefoxitin (30 Zg) disc and confirmed by inhibitor based test using boronic acid as inhibitor. A total of 16 E.coli isolates (40%) and 16 Klebsiella ...

  9. Resistance Pattern and Detection of Metallo-beta-lactamase Genes ...

    African Journals Online (AJOL)

    2018-02-23

    Feb 23, 2018 ... P. aeruginosa from clinical samples submitted to the Medical Microbiology .... 28 species of Gram-negative bacteria from more than ..... medical tourism, and work. .... aeruginosa: Mechanisms and impact on treatment.

  10. Antibiotic resistance patterns and beta-lactamase identification in ...

    African Journals Online (AJOL)

    Background. Antibiotic resistance is a growing problem worldwide. Mechanisms of resistance vary, and some can confer resistance to multiple classes of antibiotics. Objective. To characterise the antibiotic resistance profiles of Escherichia coli isolates obtained from stool samples of young rural children exposed or ...

  11. Extended-spectrum beta-lactamase producing gram negative ...

    African Journals Online (AJOL)

    African Journal of Infectious Diseases ... The rate of nosocomial infections caused by ESBL-producing GNB in Asia Pacific ... Results: In almost all parts of the country, high resistance has been observed, especially in the central part of Iran.

  12. New Delhi Metallo – beta lactamase – 1 containing

    African Journals Online (AJOL)

    raoul

    2012-02-13

    Feb 13, 2012 ... Diagnosis, Treatment and Public health concern ... resistance is because of a carbapenemase enzyme called NDM- 1 (New Delhi Metallo-betalactamase - .... The activity of these enzymes can be inhibited by Zinc chelating.

  13. 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

    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 bidirectional transmission between pets and humans, especially at household level....

  14. Chromobacterium spp. harbour Ambler class A beta-lactamases showing high identity with KPC

    DEFF Research Database (Denmark)

    Gudeta, Dereje Dadi; Bortolaia, Valeria; Jayol, Aurelie

    2016-01-01

    Objectives: The origin of KPC is unknown. The aim of this study was to detect progenitors of KPC in silico and to functionally verify their beta-lactam hydrolysis activity. Methods: The sequence of KPC-2 was used to mine the NCBI protein sequence database. The best non-KPC hits were analysed by a......-lactamases with up to 76% aa identity to KPC from distinct Chromobacterium species is highly indicative of the role played by this genus in the evolution of KPC....

  15. 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

  16. A fungal metallo-beta-lactamase necessary for biotransformation of maize phytoprotectant compounds

    Science.gov (United States)

    Xenobiotic compounds such as phytochemicals, microbial metabolites, and agrochemicals can impact the diversity and frequency of fungal species occurring in agricultural environments. Resistance to xenobiotics may allow plant pathogenic fungi to dominate the overall fungal community, with potential ...

  17. Classification of Beta-lactamases and penicillin binding proteins using ligand-centric network models.

    Directory of Open Access Journals (Sweden)

    Hakime Öztürk

    Full Text Available β-lactamase mediated antibiotic resistance is an important health issue and the discovery of new β-lactam type antibiotics or β-lactamase inhibitors is an area of intense research. Today, there are about a thousand β-lactamases due to the evolutionary pressure exerted by these ligands. While β-lactamases hydrolyse the β-lactam ring of antibiotics, rendering them ineffective, Penicillin-Binding Proteins (PBPs, which share high structural similarity with β-lactamases, also confer antibiotic resistance to their host organism by acquiring mutations that allow them to continue their participation in cell wall biosynthesis. In this paper, we propose a novel approach to include ligand sharing information for classifying and clustering β-lactamases and PBPs in an effort to elucidate the ligand induced evolution of these β-lactam binding proteins. We first present a detailed summary of the β-lactamase and PBP families in the Protein Data Bank, as well as the compounds they bind to. Then, we build two different types of networks in which the proteins are represented as nodes, and two proteins are connected by an edge with a weight that depends on the number of shared identical or similar ligands. These models are analyzed under three different edge weight settings, namely unweighted, weighted, and normalized weighted. A detailed comparison of these six networks showed that the use of ligand sharing information to cluster proteins resulted in modules comprising proteins with not only sequence similarity but also functional similarity. Consideration of ligand similarity highlighted some interactions that were not detected in the identical ligand network. Analysing the β-lactamases and PBPs using ligand-centric network models enabled the identification of novel relationships, suggesting that these models can be used to examine other protein families to obtain information on their ligand induced evolutionary paths.

  18. Extended Spectrum Beta-Lactamase Carriage State among Elderly Nursing Home Residents in Beirut

    Directory of Open Access Journals (Sweden)

    M. A. Jallad

    2015-01-01

    Full Text Available Introduction. ESBL-producing Enterobacteriaceae can cause severe infections, but they are also isolated from the stool of asymptomatic subjects. Faecal carriage of such organism is poorly understood. Methods. First phase of the study was cross-sectional with prevalence and epidemiology of ESBL faecal carriage in two nursing homes in Beirut: 57 residents in the first (NH1 and 151 residents in the second (NH2. In second phase, faecal swabs from cohort of NH1 residents were examined for carriage at six-week intervals over three-month period. Residents’ charts were reviewed to assess carriage risk factors. Results. Over 3 consecutive samplings at NH1, 81% of residents were at least one-time carriers with 50% at the first round, 60.4% at the second, and 74.5% at the last one. At NH2, 68.2% of residents were carriers. Constipation (in NH1 and antibiotic intake (in NH2 were significantly associated with higher ESBL faecal carriage while the length of stay at the nursing home (in NH2 was associated with less carriage. Conclusion. Faecal carriage of ESBL-producing Enterobacteriaceae is high among nursing home patients in Beirut. The rate of carriage changes rapidly and significantly over time either with multiple factors playing a possible role like outbreak spreading, antibiotic, and health care system exposure.

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    Tatiana Čornejová

    2015-09-01

    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, qnrS genes and virulence factors present a potential environmental health risk.

  1. Extended-Spectrum Beta-Lactamase (ESBL)–Producing Gram ...

    African Journals Online (AJOL)

    Tropical Journal of Pharmaceutical Research is indexed by Science Citation Index (SciSearch), Scopus,. International Pharmaceutical Abstract, Chemical Abstracts, Embase, Index Copernicus, EBSCO, African. Index Medicus, JournalSeek, Journal Citation Reports/Science Edition, Directory of Open Access Journals.

  2. Bovine intestinal bacteria inactivate and degrade ceftiofur and ceftriaxone with multiple beta-lactamases.

    Science.gov (United States)

    Wagner, R Doug; Johnson, Shemedia J; Cerniglia, Carl E; Erickson, Bruce D

    2011-11-01

    The veterinary cephalosporin drug ceftiofur is rapidly degraded in the bovine intestinal tract. A cylinder-plate assay was used to detect microbiologically active ceftiofur, and high-performance liquid chromatography-mass spectrometry analysis was used to quantify the amount of ceftiofur remaining after incubation with bovine intestinal anaerobic bacteria, which were isolated from colon contents or feces from 8 cattle. Ninety-six percent of the isolates were able to inactivate ceftiofur to some degree, and 54% actually degraded the drug. None of 9 fungal isolates inactivated or degraded ceftiofur. Facultative and obligate anaerobic bacterial species that inactivated or degraded ceftiofur were identified with Vitek and Biolog systems, respectively. A subset of ceftiofur degraders also degraded the chemically similar drug ceftriaxone. Most of the species of bacteria that degraded ceftiofur belonged to the genera Bacillus and Bacteroides. PCR analysis of bacterial DNA detected specific β-lactamase genes. Bacillus cereus and B. mycoides isolates produced extended-spectrum β-lactamases and metallo-β-lactamases. Seven isolates of Bacteroides spp. produced multiple β-lactamases, including possibly CepA, and metallo-β-lactamases. Isolates of Eubacterium biforme, Bifidobacterium breve, and several Clostridium spp. also produced ceftiofur-degrading β-lactamases. An agar gel overlay technique on isoelectric focusing separations of bacterial lysates showed that β-lactamase enzymes were sufficient to degrade ceftiofur. These results suggest that ceftiofur is inactivated nonenzymatically and degraded enzymatically by multiple β-lactamases from bacteria in the large intestines of cattle.

  3. Fecal microbiota transplantation against intestinal colonization by extended spectrum beta-lactamase producing Enterobacteriaceae

    NARCIS (Netherlands)

    Singh, Ramandeep; Groot, de Pieter F.; Geerlings, Suzanne E.; Hodiamont, Caspar J.; Belzer, Clara; Berge, ten Ineke J.M.; Vos, de Willem M.; Bemelman, Frederike J.; Nieuwdorp, Max

    2018-01-01

    Objective: Infections with multidrug-resistant microorganisms are associated with increased hospitalization, medication costs and mortality. Based on our fecal microbiota transplantation (FMT) experience for Clostridium difficile infection, we treated 15 patients carrying ESBL-producing

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

    DEFF Research Database (Denmark)

    Bagge, N.; Hentzer, Morten; Andersen, Jens Bo

    2004-01-01

    of increased imipenem concentrations. Ceftazidime induced the monitor system of the biofilm bacteria as well, but only bacteria in the peripheries of the microcolonies were induced in the presence of even very high concentrations. The experiments illustrate for the first time the dynamic and spatial...

  5. Molecular characteristics of extended-spectrum beta-lactamases and qnr determinants in Enterobacter species from Japan.

    Science.gov (United States)

    Kanamori, Hajime; Yano, Hisakazu; Hirakata, Yoichi; Hirotani, Ayako; Arai, Kazuaki; Endo, Shiro; Ichimura, Sadahiro; Ogawa, Miho; Shimojima, Masahiro; Aoyagi, Tetsuji; Hatta, Masumitsu; Yamada, Mitsuhiro; Gu, Yoshiaki; Tokuda, Koichi; Kunishima, Hiroyuki; Kitagawa, Miho; Kaku, Mitsuo

    2012-01-01

    The incidence of extended-spectrum β-lactamases (ESBLs) has been increasing worldwide, but screening criteria for detection of ESBLs are not standardized for AmpC-producing Enterobacteriaceae such as Enterobacter species. In this study, we investigated the prevalence of ESBLs and/or AmpC β-lactamases in Japanese clinical isolates of Enterobacter spp. and the association of plasmid-mediated quinolone resistance (PMQR) determinants with ESBL producers. A total of 364 clinical isolates of Enterobacter spp. collected throughout Japan between November 2009 and January 2010 were studied. ESBL-producing strains were assessed by the CLSI confirmatory test and the boronic acid disk test. PCR and sequencing were performed to detect CTX-M, TEM, and SHV type ESBLs and PMQR determinants. For ESBL-producing Enterobacter spp., pulsed-field gel electrophoresis (PFGE) was performed using XbaI restriction enzyme. Of the 364 isolates, 22 (6.0%) were ESBL producers. Seven isolates of Enterobacter cloacae produced CTX-M-3, followed by two isolates producing SHV-12. Two isolates of Enterobacter aerogenes produced CTX-M-2. Of the 22 ESBL producers, 21 had the AmpC enzyme, and six met the criteria for ESBL production in the boronic acid test. We found a significant association of qnrS with CTX-M-3-producing E. cloacae. The 11 ESBL-producing Enterobacter spp. possessing bla(CTX-M), bla(SHV), or bla(TEM) were divided into six unique PFGE types. This is the first report about the prevalence of qnr determinants among ESBL-producing Enterobacter spp. from Japan. Our results suggest that ESBL-producing Enterobacter spp. with qnr determinants are spreading in Japan.

  6. Detection of New Delhi Metallo-Beta-Lactamase (Encoded by blaNDM-1 ) in Enterobacter aerogenes in China.

    Science.gov (United States)

    Shen, Yong; Xiao, Wei-Qiang; Gong, Jiao-Mei; Pan, Jun; Xu, Qing-Xia

    2017-03-01

    The increase in bla NDM -1 in Enterobacteriaceae has become a major concern worldwide. In previous study, we investigated clonal dissemination and mechanisms of resistance to carbapenem in China. We carried out retrospective surveillance for bla NDM -1 among carbapenem-resistant enterobacter strains, which were isolated from patients at our hospital by bacterial strains selection, antimicrobial susceptibility testing, species identification, and molecular detection of resistance gene. We found three bla NDM -1 -positive isolates which were identified as Enterobacter aerogenes in clinical patients in China. The bla NDM -1 -positive Enterobacter aerogenes isolates were first found. It is important to mandate prudent usage of antibiotics and implement infection control measures to control the spread of these resistant bla NDM -1 -positive strains. © 2016 Wiley Periodicals, Inc.

  7. Free Energy Contribution Analysis Using Response Kernel Approximation: Insights into the Acylation Reaction of a Beta-Lactamase.

    Science.gov (United States)

    Asada, Toshio; Ando, Kanta; Bandyopadhyay, Pradipta; Koseki, Shiro

    2016-09-08

    A widely applicable free energy contribution analysis (FECA) method based on the quantum mechanical/molecular mechanical (QM/MM) approximation using response kernel approaches has been proposed to investigate the influences of environmental residues and/or atoms in the QM region on the free energy profile. This method can evaluate atomic contributions to the free energy along the reaction path including polarization effects on the QM region within a dramatically reduced computational time. The rate-limiting step in the deactivation of the β-lactam antibiotic cefalotin (CLS) by β-lactamase was studied using this method. The experimentally observed activation barrier was successfully reproduced by free energy perturbation calculations along the optimized reaction path that involved activation by the carboxylate moiety in CLS. It was found that the free energy profile in the QM region was slightly higher than the isolated energy and that two residues, Lys67 and Lys315, as well as water molecules deeply influenced the QM atoms associated with the bond alternation reaction in the acyl-enzyme intermediate. These facts suggested that the surrounding residues are favorable for the reactant complex and prevent the intermediate from being too stabilized to proceed to the following deacylation reaction. We have demonstrated that the free energy contribution analysis should be a useful method to investigate enzyme catalysis and to facilitate intelligent molecular design.

  8. 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.

  9. 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.

  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. 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...

  12. Prevalence of AmpC and other beta-lactamases in enterobacteria at a large urban university hospital in Brazil

    Science.gov (United States)

    Dias, Rubens Clayton da Silva; Borges-Neto, Armando Alves; Ferraiuoli, Giovanna Ianini D’Almeida; de-Oliveira, Márcia P.; Riley, Lee W.; Moreira, Beatriz Meurer

    2010-01-01

    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 occurrence of ESBL and pAmpC β-lactamases in a hospital where MDR enterobacterial isolates recently emerged. A total of 123 consecutive enterobacterial isolates obtained from 112 patients at a university hospital in Rio de Janeiro, Brazil during March-June 2001 were included in the study. ESBL was detected by the addition of clavulanate to cephalosporin containing disks and by double diffusion. AmpC production was evaluated by a modified tridimensional test and a modified Hodge test. The presence of plasmid-mediated ampC β-lactamase genes was evaluated by multiplex-PCR. Sixty-five (53%) of 123 enterobacterial isolates were MDR, obtained from 56 patients. ESBL production was detected in 35 isolates; 5 clonal E. coli isolates exhibited high levels of chromosomal AmpC and ESBL production. However, no isolates contained pAmpC genes. Infection or colonization by MDR enterobacteria was not associated with any predominant resistant clones. A large proportion of hospital infections caused by ESBL-producing enterobacteria identified during the study period were due to sporadic infections rather than undetected outbreaks. This observation emphasizes the need to improve our detection methods for ESBL- and AmpC-producing organisms in hospitals where extended-spectrum cephalosporins are in wide use. PMID:17900845

  13. 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.

  14. Rectal carriage of extended-spectrum beta-lactamase-producing gram-negative bacilli in community settings in Madagascar.

    Directory of Open Access Journals (Sweden)

    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.

  15. Occurrence and sensitivity profile of extended spectrum beta-lactamase-producing Enterobacteriaceae at a tertiary hospital in Southern Brazil

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    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.

  16. Molecular characterization and antimicrobial susceptibility profile of New Delhi metallo-beta-lactamase-1-producing Escherichia coli among hospitalized patients

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    Anjali Agarwal

    2018-01-01

    CONCLUSION: There is an increased prevalence of NDM-1 gene-producing E. coli isolates. These carbapenemase-producing isolates are more resistant to other group of antibiotics (aminoglycosides, fluoroquinolones along with β-lactam group. Early detection of bla NDM-1 gene can help in choosing the effective treatment options for hospitalized patients in time, thereby reducing the risk of mortality.

  17. Nosocomial extended-spectrum beta-lactamase-producing Klebsiella pneumoniae bacteremia in hemodialysis patients and the implications for antibiotic therapy

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    Chih-Chao Yang

    2014-11-01

    Conclusions: In accordance with our previous study, our results have demonstrated the inferiority of flomoxef to carbapenems in the treatment of HD access-related ESBL-Kp bacteremia and provide an insight into the possibility of using ertapenem rather than flomoxef as an initial or de-escalating therapy for infections caused by ESBL-producing bacteria.

  18. Prevalence of Multiple Drug Resistant Clinical Isolates of Extended-Spectrum Beta-Lactamase Producing Enterobacteriaceae in Southeast Iran

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    Shahla Mansouri

    2010-06-01

    Full Text Available AbstractBackground: Multidrug resistance and production of extendedspectrum β-lactamases (ESBLs by enteric gramnegativerods in hospitals and community continue to beworsened. We aimed to characterize the multidrug resistanceand determine the prevalence of ESBL production by clinicalisolates of Enterobacteriaceae in southeast Iran.Methods: Gram-negative bacteria isolated from clinical samplesof hospital inpatients and outpatients from three hospitalsin southeast Iran were tested for susceptibility to 10commonly used antimicrobials. For 500 isolates whichshowed resistance to ≥3 antibiotics from different classes,minimum inhibitory concentration, and prevalence of ESBLproduction were determined by agar dilution and double discsynergy method respectively. The isolated bacterial specieswere compared in respect of antibacterial resistance, ESBLproduction, patients' gender, hospital ward, and type ofspecimen.Results: The most frequent resistance was to trimethoprim/sulfamethoxazole, amoxicillin, and tetracycline. Imipenemwith 99.8% and ceftizoxime with 83% susceptibility were themost active agents. A total of 53.8% of isolates expressedESBL production. Escherichia coli and Klebsiella pneumoniaewere most common in outpatients, and inpatients samplesrespectively. Higher rate of resistance to most antibacterialagents and ESBL production was found in samples ofinpatients.Conclusion: The present study showed high prevalence ofESBL-producing Enterobacteriaceae especially in the patientsadmitted to hospital. Infection control strategy with continuousresistance surveillance is essential to monitor in vitro susceptibilityto antibacterial agents currently used in clinicalpractice. Determination of the type of involved ESBL enzymesis important for a better antimicrobial control and empiricaltherapy of critically ill patients in hospitals.Iran J Med Sci 2010; 35(2: 101-108.

  19. Multiplex PCR Study of Plasmid-Mediated AmpC Beta-Lactamases Genes in Clinical Isolates of Escherichia coli

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    Maryam Dehghani

    2017-02-01

    Full Text Available Background:   AmpC β-lactamases are important cephalosporinases chromosomally encoded in many of Enterobacteriaceae and a few other organisms where they mediate resistance to cephalothin, cefazolin, cefoxitin and penicillins. The six different families of plasmid-mediated AmpC β-lactamases have been described, but no phenotypic test can discriminate among them. AmpC multiplex PCR has been successfully used to discriminate plasmid-mediated ampC specific families in organisms such as Klebsiella pneumonia and Escherichia coli. The aim of this study was to indicate the prevalence of AmpC β-lactamase genes by specifically designed primers through PCR test.Methods:   243 total clinical urine samples were collected, and 227 isolates were identified as Escherichia coli based on standard biochemical tests. Subsequently, the isolates were screened by disc diffusion and combined disc test for β-lactamase production. Resistant isolates were evaluated by PCR for ampC family determination. Results:  Antibiotic resistance pattern were observed as follows: cefepime (%25, ceftazidime (%31, ceftriaxone (%37, cefotaxime (%38. The ratio of isolates was detected as ESBLs and AmpC producers were 34% and 5.2%, respectively. PCR performed on 12 selected isolates via phenotypic tests and the results revealed that among 12 isolates, 11 contained blaCMY-42. Conclusion:  Unfortunately, antibiotic resistance has become an increasingly critical problem in many countries like Iran and occurrence of isolates co-expressing AmpC-β-lactamases and ESBLs can create serious problems in the future. As antibiotic options in the treatment of AmpC β-lactamases and ESBLs producing organisms are extremely limited, molecular screening by laboratories is suggested to reduce the risk of therapeutic defeat.

  20. High diversity of beta-lactamases in the General Hospital Vienna verified by whole genome sequencing and statistical analysis.

    Science.gov (United States)

    Barišić, Ivan; Mitteregger, Dieter; Hirschl, Alexander M; Noehammer, Christa; Wiesinger-Mayr, Herbert

    2014-10-01

    The detailed analysis of antibiotic resistance mechanisms is essential for understanding the underlying evolutionary processes, the implementation of appropriate intervention strategies and to guarantee efficient treatment options. In the present study, 110 β-lactam-resistant, clinical isolates of Enterobacteriaceae sampled in 2011 in one of Europe's largest hospitals, the General Hospital Vienna, were screened for the presence of 31 β-lactamase genes. Twenty of those isolates were selected for whole genome sequencing (WGS). In addition, the number of β-lactamase genes was estimated using biostatistical models. The carbapenemase genes blaKPC-2, blaKPC-3, and blaVIM-4 were identified in carbapenem-resistant and intermediate susceptible isolates, blaOXA-72 in an extended-spectrum β-lactamase (ESBL)-positive one. Furthermore, the observed high prevalence of the acquired blaDHA-1 and blaCMY AmpC β-lactamase genes (70%) in phenotypically AmpC-positive isolates is alarming due to their capability to become carbapenem-resistant upon changes in membrane permeability. The statistical analyses revealed that approximately 55% of all β-lactamase genes present in the General Hospital Vienna were detected by this study. In summary, this work gives a very detailed picture on the disseminated β-lactamases and other resistance genes in one of Europe's largest hospitals. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Cephem Potentiation by Inactivation of Nonessential Genes Involved in Cell Wall Biogenesis of beta-Lactamase-Producing Escherichia coli

    DEFF Research Database (Denmark)

    Baker, Kristin R.; Sigurdardottir, Helga Høeg; Jana, Bimal

    2017-01-01

    Reversal of antimicrobial resistance is an appealing and largely unexplored strategy in drug discovery. The objective of this study was to identify potential targets for “helper” drugs reversing cephem resistance in Escherichia coli strains producing β-lactamases. A CMY-2-encoding plasmid...... was transferred by conjugation to seven isogenic deletion mutants exhibiting cephem hypersusceptibility. The effect of each mutation was evaluated by comparing the MICs in the wild type and the mutant harboring the same plasmid. Mutation of two genes encoding proteins involved in cell wall biosynthesis, dap...... for all three drugs. Individual deletion of dapF and mrcB in a clinical isolate of CTX-M-15-producing E. coli sequence type 131 (ST131) resulted in partial reversal of ceftazidime and cefepime resistance but did not reduce MICs below susceptibility breakpoints. Growth curve analysis indicated no fitness...

  2. Multidrug-Resistant and Extended Spectrum Beta-Lactamase-Producing Escherichia coli in Dutch Surface Water and Wastewater.

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    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.

  3. Multi drug resistance and Extended Spectrum Beta Lactamases in clinical isolates of Shigella: A study from New Delhi, India.

    Science.gov (United States)

    Aggarwal, Prabhav; Uppal, Beena; Ghosh, Roumi; Krishna Prakash, S; Chakravarti, Anita; Jha, Arun Kumar; Rajeshwari, Krishnan

    2016-01-01

    Shigella is an important cause of gastroenteritis in local Indian population, as well as of traveler's diarrhea in the international visitors to India. These patients often require appropriate antimicrobial therapy; however, rapid development of antimicrobial resistance poses a major hurdle in achieving this goal. A prospective study was conducted during 2009-12 in New Delhi, India, including 6339 stool samples from gastroenteritis patients. 121 Shigella strains were identified on the basis of colony morphology, biochemical reactions, serotyping and ipaH gene based PCR. Antimicrobial susceptibility testing by disc diffusion, MIC determination by Vitek(®) 2 and phenotypic tests for ESBL/AmpC production were done. Nineteen percent strains (23/121) were found to be resistant to third generation cephalosporins and all were phenotypically confirmed to be ESBL producers; one strain was positive for AmpC. ESBL producing strains were also found to be significantly more resistant (p Shigella is a matter of concern for the local population as well as international travelers. Therefore, better national level antimicrobial management programs are the priority needs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. 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......-producing E. coli to cefotaxime and to determine whether the response depended on the growth phase of the bacterium and the concentration of antibiotic. Results: Two antibiotic resistant strains carrying bla(CTX-M-1) on the chromosome and on an IncI1 plasmid and three sensitive strains were used...... to cefotaxime. The filament formation was restricted to early growth phases and the time the cells grew as filaments was antibiotic concentration dependent. This indicates that antibiotic resistant E. coli undergo the same morphological changes as sensitive bacteria in the presence of beta-lactam antibiotic...

  5. Antimicrobial Resistance status and prevalence rates of Extended Spectrum Beta-Lactamase (ESBL producers isolated from a mixed human population.

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    Ruth A. Afunwa

    2011-05-01

    Full Text Available Owing to the increasing epidemiological and therapeutic challenges associated with infections due to ESBL producers, ESBL prevalence rate among some bacteria isolates from healthy and non-healthy human population in a metropolitan Nigerian setting was evaluated.A total of one hundred and forty-five (145 bacteria strains were isolated from a total of four hundred and sixty (460 samples collected from urine, wound, throat and anal swabs of 220 healthy volunteers in the community and from 240 patients in 2 secondary and 2 tertiary hospitals (altogether, 4 in Enugu metropolis. The presumptive confirmatory test used for ESBL detection was the Double Disc Synergy Test (DDST method. Conjugation and plasmid curing studies were also done for resistance factor determination.Of the 145 isolates, 20 were ESBL producers with 35% of these ESBL producers being of community origin and 65% from hospitals. This translates to 4.8% and 9% incidences (comparably higher than established prevalence of 4.4% and 7.5 respectively for community and hospital infections respectively. The ESBL isolates showed high resistance to tetracycline, gentamicin, pefloxacin, ceftriaxone, cefuroxime, ciprofloxacin and Augmentin® (Amoxicilin and clavulanic acid combination. Conjugation studies for Resistance plasmid transfer showed non-transference of resistance determinants between the ESBL transconjugants and recipient strains. Correspondingly, the plasmid curing studies revealed that the acridine orange could not effect a cure on the isolates as they still retained high resistance to the antibiotics after the treatment.This study confirms the growing incidences/pool of ESBL strains in Nigeria and call for widespread and continuous monitoring towards an effective management of the potential therapeutic hurdle posed by this trend.

  6. in vitro effectiveness of commercial bacteriophage cocktails on diverse extended spectrum beta-lactamase (ESBL producing Escherichia coli strains

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    Aycan Gundogdu

    2016-11-01

    Full Text Available The objective of this study is to determine the in vitro susceptibility of Georgian bacteriophage cocktails on multi-drug resistant extended-spectrum β-lactamase producing Escherichia coli (ESBL-EC isolated from patients' blood and urine cultures. 615 E. coli isolates were included in this study. PhP-typing and phylogenetic grouping were used for the typing. Antimicrobial resistance profiles and ESBL production of all isolates were confirmed according to CLSI criteria. The activities of four bacteriophage cocktails (Enko-phage, SES-bacteriophage, Pyo-bacteriophage and Intesti-bacteriophage were determined against 142 ESBL- EC using in vitro spot tests. According to this, Enko-phage were active against 87.3% of the tested strains while that ratio was 81.7% for intesti-bacteriophage, 81.7% for Pyo-bacteriophage and 59.2% for SES-bacteriophage cocktails. Based on the contingency tests, the phage cocktails were observed to be statistically significantly (p<0.001 more effective on ESBL-EC strains belonging to phylogenetic groups D and B2. The employed phage cocktails were found to be affective against all tested resistant types. These results are promising especially for the infections that are caused by multi-drug resistant pathogens that are difficult to treat. As this is a preliminary step to the potential clinical trials to be designed for the country, in vitro confirmation of their success on a multi-drug-resistant ESBL-EC collection should be accepted as an initial action, which is encouraging to consider clinical trials of phage therapy especially in countries which are not introduce phage therapy.

  7. RELATION BETWEEN BETA-LACTAMASE-PRODUCING BACTERIA AND PATIENT CHARACTERISTICS IN CHROMIC OBSTRUCTIVE PULMONARY-DISEASE (COPD)

    NARCIS (Netherlands)

    SPORTEL, JH; KOETER, GH; VANALTENA, R; LOWENBERG, A; BOERSMA, WG

    Background - In addition to bronchodilator and anti-inflammatory therapy, exacerbations in patients with chronic obstructive pulmonary disease (COPD) are often treated with antibiotics. Haemophilus influenzae and Moraxella (Branhamella) catarrhalis, two important respiratory pathogens, may produce

  8. Sequence-function-stability relationships in proteins from datasets of functionally annotated variants: The case of TEM beta-lactamases

    NARCIS (Netherlands)

    Abriata, L.A.; Salverda, M.L.M.; Tomatis, P.E.

    2012-01-01

    A dataset of TEM lactamase variants with different substrate and inhibition profiles was compiled and analyzed. Trends show that loops are the main evolvable regions in these enzymes, gradually accumulating mutations to generate increasingly complex functions. Notably, many mutations present in

  9. 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.

    In this study a mathematical model for the spread of ESBL resistant E.coli among patients in a hospital and the surrounding catchment population has been introduced and used to described prevalence data from the Netherlands. Several statistical methods have been applied to estimate the model...

  10. Acinetobacter baumannii: Epidemiological and Beta-Lactamase Data From Two Tertiary Academic Hospitals in Tshwane, South Africa

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    Michelle Lowe

    2018-06-01

    Full Text Available Acinetobacter baumannii is an opportunistic pathogen that is increasingly responsible for hospital-acquired infections. The increasing prevalence of carbapenem resistant A. baumannii has left clinicians with limited treatment options. Last line antimicrobials (i.e., polymyxins and glycylcyclines are often used as treatment options. The aim of this study was to determine the prevalence of selected β-lactamase genes from A. baumannii isolates obtained from patients with hospital-acquired infections and to determine the genetic relationship and epidemiological profiles among clinical A. baumannii isolates collected from two tertiary academic hospitals in the Tshwane region, South Africa (SA. Multiplex-PCR (M-PCR assays were performed to detect selected resistance genes. The collected isolates’ genetic relatedness was determined by using pulsed field gel electrophoresis (PFGE and multilocus sequence typing (MLST. The acquired oxacillinase (OXA genes, notably blaOXA-23-like were prevalent in the A. baumannii isolates. The M-PCR assays showed that the isolates collected from hospital A contained the OXA-23-like (96%; n = 69/72 genes and the isolates collected from hospital B contained the OXA-23-like (91%; n = 63/69 and OXA-58-like (4%; n = 3/69 genes. Colistin resistance was found in 1% of the isolates (n = 2/141 and tigecycline intermediate resistance was found in 6% of the isolates (n = 8/141. The A. baumannii isolates were genetically diverse. Molecular epidemiological data showed that specific sequence types (STs (ST106, ST229, ST258 and ST208 were established in both hospitals, while ST848 was established in hospital A and ST502, ST339 and the novel ST1552 were established in hospital B. ST848 (established in hospital A was predominately detected in ICU wards whereas ST208, ST339 and the novel ST1552 (established in hospital B were detected in ICUs and the general wards. The origin of the A. baumannii isolates in the hospitals may be due to the dissemination and adaptation of a diverse group of successful clones. Poor infection control and prevention strategies and possibly the overuse of antimicrobials contributed to the establishment of these A. baumannii clones in the studied hospitals.

  11. 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 : antibiotics * 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

  12. 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.

  13. Prevalence, risk factors, and impact on clinical outcome of extended-spectrum beta-lactamase-producing Escherichia coli bacteraemia: a five-year study

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    B. Denis

    2015-10-01

    Conclusion: The prevalence of ESBL-EC bacteraemia has been increasing dramatically. Previous colonization with ESBL-EC was a strong risk factor for ESBL-EC bacteraemia. More inadequate initial antimicrobial therapy was noted in the ESBL-EC group, but mortality and length of hospital stay were not significantly different from those of patients with non-ESBL-EC bacteraemia.

  14. Carriage of extended-spectrum beta-lactamase-plasmids does not reduce fitness but enhances virulence in some strains of pandemic E. coli lineages

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    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.

  15. 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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Emergence of New Delhi metallo-beta-lactamase 1 and other carbapenemase-producing Acinetobacter calcoaceticus-baumannii complex among patients in hospitals in Ha Noi, Viet Nam.

    Science.gov (United States)

    Tran, D N; Tran, H H; Matsui, M; Suzuki, M; Suzuki, S; Shibayama, K; Pham, T D; Van Phuong, T T; Dang, D A; Trinh, H S; Loan, C T; Nga, L T V; van Doorn, H R; Wertheim, H F L

    2017-02-01

    Acinetobacter baumannii is an important cause of multidrug-resistant hospital acquired infections in the world. Here, we investigate the presence of NDM-1 and other carbapenemases among carbapenem-resistant A. baumannii isolated between August 2010 and December 2014 from three large hospitals in Hanoi, Vietnam. We identified 23/582 isolates (4 %) (11 from hospital A, five from hospital B, and seven from hospital C) that were NDM-1 positive, and among them 18 carried additional carbapenemase genes, including seven isolates carrying NDM-1, IMP-1, and OXA-58 with high MICs for carbapenems. Genotyping indicated that NDM-1 carrying A. baumannii have expanded clonally in these hospitals. Five new STs (ST1135, ST1136, ST1137, ST1138, and ST1139) were identified. One isolate carried NDM-1 on a plasmid belonging to the N-repA replicon type; no NDM-1-positive plasmids were identified in the other isolates. We have shown the extent of the carbapenem resistance and the local clonal spread of A. baumannii carrying NDM-1 in these hospitals; coexistence of NDM-1 and IMP-1 is reported for the first time from Vietnam here, and this will further seriously limit future therapeutic options.

  17. Emergence of New Delhi metallo-beta-lactamase 1 and other carbapenemase-producing Acinetobacter calcoaceticus-baumannii complex among patients in hospitals in Ha Noi, Viet Nam

    NARCIS (Netherlands)

    Tran, D.N.; Tran, H.H.; Matsui, M.; Suzuki, M.; Suzuki, S.; Shibayama, K.; Pham, T.D.; Phuong, T.T. Van; Dang, D.A.; Trinh, H.S.; Loan, C.T.; Nga, L.T.; Doorn, H.R. van; Wertheim, H.F.L.

    2017-01-01

    Acinetobacter baumannii is an important cause of multidrug-resistant hospital acquired infections in the world. Here, we investigate the presence of NDM-1 and other carbapenemases among carbapenem-resistant A. baumannii isolated between August 2010 and December 2014 from three large hospitals in

  18. Clinical profiles of patients colonized or infected with extended-spectrum beta-lactamase producing Enterobacteriaceae isolates: a 20 month retrospective study at a Belgian University Hospital

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    Jamart Jacques

    2011-01-01

    Full Text Available Abstract Background Description of the clinical pictures of patients colonized or infected by ESBL-producing Enterobacteriaceae isolates and admitted to hospital are rather scarce in Europe. However, a better delineation of the clinical patterns associated with the carriage of ESBL-producing isolates may allow healthcare providers to identify more rapidly at risk patients. This matter is of particular concern because of the growing proportion of ESBL-producing Enterobacteriaceae species isolates worldwide. Methods We undertook a descriptive analysis of 114 consecutive patients in whom ESBL-producing Enterobacteriaceae isolates were collected from clinical specimens over a 20-month period. Clinical data were obtained through retrospective analysis of medical record charts. Microbiological cultures were carried out by standard laboratory methods. Results The proportion of ESBL-producing Enterobacteriaceae strains after exclusion of duplicate isolates was 4.5% and the incidence rate was 4.3 cases/1000 patients admitted. Healthcare-associated acquisition was important (n = 104 while community-acquisition was less frequently found (n = 10. Among the former group, two-thirds of the patients were aged over 65 years and 24% of these were living in nursing homes. Sixty-eight (65% of the patients with healthcare-associated ESBL, were considered clinically infected. In this group, the number and severity of co-morbidities was high, particularly including diabetes mellitus and chronic renal insufficiency. Other known risk factors for ESBL colonization or infection such as prior antibiotic exposure, urinary catheter or previous hospitalisation were also often found. The four main diagnostic categories were: urinary tract infections, lower respiratory tract infections, septicaemia and intra-abdominal infections. For hospitalized patients, the median hospital length of stay was 23 days and the average mortality rate during hospitalization was 13% (Confidence Interval 95%: 7-19. Escherichia coli, by far, accounted as the most common ESBL-producing Enterobacteriaceae species (77/114; [68%] while CTX-M-1 group was by far the most prevalent ESBL enzyme (n = 56. Conclusion In this retrospective study, the clinical profiles of patients carrying healthcare-associated ESBL-producing Enterobacteriacae is characterized by a high prevalence rate of several major co-morbidities and potential known risk factors. Both, the length of hospital stay and overall hospital mortality rates were particularly high. A prospective case-control matched study should be designed and performed in order to control for possible inclusion bias.

  19. Insight into the effect of inhibitor resistant S130G mutant on physico-chemical properties of SHV type beta-lactamase: a molecular dynamics study.

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    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. 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)

    Chen Hao; Shu Weiqun; Chang Xiaosong; Chen Jian; Guo Yebin; Tan Yao

    2010-01-01

    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 bla 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.

  1. Seawater is a reservoir of multi-resistant Escherichia coli, including strains hosting plasmid-mediated quinolones resistance and extended-spectrum beta-lactamases genes.

    Science.gov (United States)

    Alves, Marta S; Pereira, Anabela; Araújo, Susana M; Castro, Bruno B; Correia, António C M; Henriques, Isabel

    2014-01-01

    The aim of this study was to examine antibiotic resistance (AR) dissemination in coastal water, considering the contribution of different sources of fecal 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 fecal contamination: human-derived sewage and seagull feces. 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 feces isolates, except for last-line antibiotics used in human medicine. Multi-resistance rates in isolates from sewage and seagull feces (29 and 32%) were lower than in isolates from seawater (39%). Seawater AR profiles were similar to those from seagull feces and differed significantly from sewage AR profiles. Nucleotide sequences matching resistance genes bla TEM, 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 (bla CTX-M-1 and bla SHV-12) and seagull feces (bla CMY-2). Plasmid-mediated determinants of resistance to quinolones were found: qnrS1 in all sources and qnrB19 in seawater and seagull feces. 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 fecal pollution. The repertoire of resistance genes covers antibiotics critically important for humans, a potential risk for human health.

  2. Identification of DHA-23, a Novel Plasmid-mediated and Inducible AmpC beta-Lactamase from Enterobacteriaceae in Northern Taiwan

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    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.

  3. [Genetic diversity of extraintestinal Escherichia coli strains producers of beta-lactamases TEM, SHV and CTX-M associated with healthcare].

    Science.gov (United States)

    Varela, Yasmin; Millán, Beatriz; Araque, María

    2017-06-01

    There are few reports from Venezuela describing the genetic basis that sustains the pathogenic potential and phylogenetics of Escherichia coli extraintestinal strains isolated in health care units. To establish the genetic diversity of extraintestinal E. coli strains producers of betalactamases TEM, SHV and CTX-M associated with healthcare. We studied a collection of 12 strains of extraintestinal E. coli with diminished sensitivity to broad-spectrum cephalosporins. Antimicrobial susceptibility was determined by minimum inhibitory concentration. We determined the phylogenetic groups, virulence factors and genes encoding antimicrobial resistance using PCR, and clonal characterization by repetitive element palindromic-PCR rep-PCR. All strains showed resistance to cephalosporins and joint resistance to quinolones and aminoglycosides. The phylogenetic distribution showed that the A and B1 groups were the most frequent, followed by D and B2. We found all the virulence factors analyzed in the B2 group, and fimH gene was the most frequent among them. We found blaCTX-M in all strains,with a higher prevalence of blaCTX-M-8; two of these strains showed coproduction of blaCTX-M-9 and were genetically identified as blaCTXM-65 and blaCTX-M-147 by sequencing. The strains under study showed genetic diversity, hosting a variety of virulence genes, as well as antimicrobial resistance with no particular phylogroup prevalence. This is the first report of blaCTX-M alleles in Venezuela and in the world associated to non-genetically related strains isolated in health care units, a situation that deserves attention, as well as the rationalization of antimicrobials use.

  4. Detection of cfxA2, cfxA3, and cfxA6 genes in beta-lactamase producing oral anaerobes

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    Buhle BINTA

    2016-04-01

    Full Text Available ABSTRACT Purpose The aim of this study was to identify β-lactamase-producing oral anaerobic bacteria and screen them for the presence of cfxA and BlaTEM genes that are responsible for β-lactamase production and resistance to β-lactam antibiotics. Material and Methods Periodontal pocket debris samples were collected from 48 patients with chronic periodontitis and anaerobically cultured on blood agar plates with and without β-lactam antibiotics. Presumptive β-lactamase-producing isolates were evaluated for definite β-lactamase production using the nitrocefin slide method and identified using the API Rapid 32A system. Antimicrobial susceptibility was performed using disc diffusion and microbroth dilution tests as described by CLSI Methods. Isolates were screened for the presence of the β-lactamase-TEM (BlaTEM and β-lactamase-cfxA genes using Polymerase Chain Reaction (PCR. Amplified PCR products were sequenced and the cfxA gene was characterized using Genbank databases. Results Seventy five percent of patients carried two species of β-lactamase-producing anaerobic bacteria that comprised 9.4% of the total number of cultivable bacteria. Fifty one percent of β-lactamase-producing strains mainly Prevotella, Porphyromonas, and Bacteroides carried the cfxA gene, whereas none of them carried blaTEM. Further characterization of the cfxA gene showed that 76.7% of these strains carried the cfxA2 gene, 14% carried cfxA3, and 9.3% carried cfxA6. The cfxA6 gene was present in three Prevotella spp. and in one Porphyromonas spp. Strains containing cfxA genes (56% were resistant to the β-lactam antibiotics. Conclusion This study indicates that there is a high prevalence of the cfxA gene in β-lactamase-producing anaerobic oral bacteria, which may lead to drug resistance and treatment failure.

  5. 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.

  6. Febrile urinary-tract infection due to extended-spectrum beta-lactamase-producing Enterobacteriaceae in children: A French prospective multicenter study.

    Science.gov (United States)

    Madhi, Fouad; Jung, Camille; Timsit, Sandra; Levy, Corinne; Biscardi, Sandra; Lorrot, Mathie; Grimprel, Emmanuel; Hees, Laure; Craiu, Irina; Galerne, Aurelien; Dubos, François; Cixous, Emmanuel; Hentgen, Véronique; Béchet, Stéphane; Bonacorsi, Stéphane; Cohen, Robert

    2018-01-01

    To assess the management of febrile urinary-tract infection (FUTIs) due to extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) in children, the Pediatric Infectious Diseases Group of the French Pediatric Society set up an active surveillance network in pediatric centers across France in 2014. We prospectively analysed data from 2014 to 2016 for all children < 18 years old who received antibiotic treatment for FUTI due to ESBL-E in 24 pediatric centers. Baseline demographic, clinical features, microbiological data and antimicrobials prescribed were collected. 301 children were enrolled in this study. The median age was 1 year (IQR 0.02-17.9) and 44.5% were male. These infections occurred in children with history of UTIs (27.3%) and urinary malformations (32.6%). Recent antibiotic use was the main associated factor for FUTIs due to ESBL-E, followed by a previous hospitalization and travel history. Before drug susceptibility testing (DST), third-generation cephalosporins (3GC) PO/IV were the most-prescribed antibiotics (75.5%). Only 13% and 24% of children received amikacine alone for empirical or definitive therapy, respectively, whereas 88.7% of children had isolates susceptible to amikacin. In all, 23.2% of children received carbapenems in empirical and/or definitive therapy. Cotrimoxazole (24.5%), ciprofloxacin (15.6%) and non-orthodox clavulanate-cefixime combination (31.3%) were the most frequently prescribed oral options after obtaining the DST. The time to apyrexia and length of hospital stay did not differ with or without effective empirical therapy. We believe that amikacin should increasingly take on a key role in the choice of definitive therapy of FUTI due to ESBL-E in children by avoiding the use of carbapenems.

  7. Impact of empirical treatment in extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella spp. bacteremia. A multicentric cohort study

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    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.

  8. Antibiotic Trapping by Plasmid-Encoded CMY-2 beta-Lactamase Combined with Reduced Outer Membrane Permeability as a Mechanism of Carbapenem Resistance in Escherichia coli

    NARCIS (Netherlands)

    Goessens, W.H.F.; van der Bij, A.K.; van Boxtel, R.; Pitout, J.D.D.; van Ulsen, J.P.; Melles, D.C.; Tommassen, J.

    2013-01-01

    A liver transplant patient was admitted with cholangitis, for which meropenem therapy was started. Initial cultures showed a carbapenem-susceptible (CS) Escherichia coli strain, but during admission, a carbapenem-resistant (CR) E. coli strain was isolated. Analysis of the outer membrane protein

  9. PREVALENCE AND SUSCEPTIBILITY OF EXTENDED SPECTRUM BETA-LACTAMASES IN URINARY ISOLATES OF ESCHERICHIA COLI IN A TERTIARY CARE HOSPITAL, CHENNAI-SOUTH INDIA

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    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.

  10. Prevalence and risk factors for extended-spectrum beta-lactamase or AmpC-producing Escherichia coli in organic dairy herds in the Netherlands

    NARCIS (Netherlands)

    Santman - Berends, Inge; Gonggrijp, M A; Heuvelink, A E; Velthuis, A; Lam, T J G M; van Schaik, Gerdien; Hage, J. J.

    2017-01-01

    Extended-spectrum β-lactamase and AmpC-producing Escherichia coli (ESBL/AmpC) are an emerging problem and are hypothesized to be associated with antimicrobial use (AMU), and more specifically with the use of third- and fourth-generation cephalosporins. Whether ESBL/AmpC also occur in organic dairy

  11. Detection of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in vegetables, soil and water of the farm environment in Tunisia.

    Science.gov (United States)

    Ben Said, Leila; Jouini, Ahlem; Klibi, Naouel; Dziri, Raoudha; Alonso, Carla Andrea; Boudabous, Abdellatif; Ben Slama, Karim; Torres, Carmen

    2015-06-16

    One-hundred-nine samples of 18 different farms (49 of food-vegetables, 41 of soil and 19 of irrigation water) and 45 vegetable food samples of 13 markets were collected in Tunisia. These samples were inoculated in MacConkey agar plates supplemented with cefotaxime (2 μg/ml). ESBL-producing Enterobacteriaceae (ESBL-Eb) were detected in 10 of the 109 farm samples (vegetables, 8.2%; soil, 7.3%; water, 15.8%), and in 4 of 45 vegetables of markets (8.9%), recovering 15 ESBL-Eb. Isolates and ESBL genes detected were: Escherichia coli (n=8: 5 blaCTX-M-1, 2 blaCTX-M-15 and one blaCTX-M-14), Citrobacter freundii (n=4: 3 blaCTX-M-15 and one blaSHV-12), Enterobacter hormaechei (n=2: 2 blaCTX-M-15) and Klebsiella pneumoniae (n=1, blaCTX-M-15). The ISEcp1 sequence was found upstream of blaCTX-M genes in 13 of 14 strains (in three cases truncated by IS5), and orf477 or IS903 downstream. Class 1 integrons were detected in five strains and contained two gene cassette arrangements (dfrA17-aadA5 and aadA1). Most isolates tested showed a multiresistant phenotype. All blaCTX-M-15-positive strains carried the aac(6')-1b-cr gene, that affects to amikacin-tobramycin-kanamycin-ciprofloxacin. Five ESBL-Eb strains carried genes of the qnr family. The 8 ESBL-positive E. coli isolates were typed as: ST58/B1 (n=3) and ST117/D, ST131/B2, ST10/A, ST23/A, and the new ST3496/D (one strain, each). From 1-2 plasmids were detected in all ESBL-positive E. coli isolates (63-179 kb). The ESBL genes were transferred by conjugation in 4 blaCTX-M-1-positive E. coli strains, and transconjugants acquired a 97 kb IncI1 plasmid. ESBL-Eb isolates are frequently disseminated in vegetable farms and potentially could be transmitted to humans through the food chain. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. High abundance and diversity of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in faeces and tonsils of pigs at slaughter.

    Science.gov (United States)

    Van Damme, I; Garcia-Graells, C; Biasino, W; Gowda, T; Botteldoorn, N; De Zutter, L

    2017-09-01

    This cross-sectional study investigates the abundance of cefotaxime-resistant Escherichia coli (CREC) in the faeces and tonsils of 96 pigs during slaughter. Moreover, different isolates from a selected number of pigs were tested to study the diversity of bla ESBL genes within E. coli isolates from one pig. Cefotaxime-resistant bacteria (based on enumeration results on MacConkey agar supplemented with 1mg/L cefotaxime) were found in the faeces of 77 pigs (80%; 95% CI: 70-87%) and the tonsils of 91 pigs (95%; 95% CI: 88%-98%). Cefotaxime-resistant E. coli (based on enumeration results on Tryptone Bile X-glucuronide agar supplemented with 1mg/L cefotaxime) were detected in 72 faecal samples (75%; 95% CI: 64-83%) and 45 tonsil samples (47%; 95% CI: 35-59%), in numbers up to 5.5 and 5.6log 10 CFU/g, respectively. On average, around 1/10,000 E. coli in both faeces and tonsils were cefotaxime-resistant, though large variations were observed between pigs. Within one sample, CREC isolates with up to five different combinations of ESBL genes were observed. In three out of 16 faecal samples and six out of 14 tonsil samples, only one ESBL gene profile was found. The high numbers of CREC that are occasionally found in the faeces and tonsils of pigs during slaughter may represent an important source of contamination of carcasses and subsequently pork. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Phenotypic and molecular detection of BLACTX-M gene extended-spectrum beta-lactamases in escherichia coli and klebsiella pneumoniae of north sumatera isolates

    Science.gov (United States)

    Hasibuan, Mirzan; Suryanto, Dwi; Lia Kusumawati, R.

    2018-03-01

    The application of antibiotics expanded-spectrum third-generation cephalosporin for the treatment of infectious diseases in hospitals is known contribute to increasing resistance due to the presence of the blaCTX-M gene in the bacteria producing ESBLs. This study was aimed to detect ESBLs, isolate phenotype and blaCTX-M genes on Escherichia coli and Klebsiella pneumoniae collected from H. Adam Malik Central Hospital. Phenotypes of the bacterial were detection using Vitek two compact, while the blaCTX-M genes were detection using polymerase chain reaction technique. The results showed that 85 (100%) isolates were ESBLs consisted of 41(48%) of Escherichia coli, and 44 (52%) of Klebsiella pneumoniae, respectively. blaCTX-M genes were detection in 62 (72.94%) of the isolates which 31 (36.47%) were Escherichia coli, and 31 (36.47%) of the isolates were Klebsiella pneumoniae, respectively. This study indicates the high prevalence of blaCTX-M genes in Escherichia coli and Klebsiella pneumoniea causing bacterial antibiotic resistance.

  14. Biosynthesis of ketomycin. (II) biomimetic model for beta-lactamase catalysis: host-guest interactions in cyclodextrin-penicillin inclusion complex

    International Nuclear Information System (INIS)

    Mak, H.W.

    1986-01-01

    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- 14 C]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 1 HNMR 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

  15. 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)

    Marta S. Alves

    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.

  16. 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.

  17. Phenotypic and Molecular Characterisation of Extended-Spectrum Beta-Lactamase Producing Escherichia coli Obtained from Animal Fecal Samples in Ado Ekiti, Nigeria

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    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.

  18. Evaluation of the HB&L carbapenemase and extended spectrum beta lactamase-AmpC automated screening kits for the rapid detection of resistant Enterobacteriaceae in rectal swabs

    Directory of Open Access Journals (Sweden)

    Sara Marani

    2017-03-01

    Full Text Available Background. In the past two decades, a rapid increase of infections due to multidrug-resistant Enterobacteriaceae was reported worldwide, including in Italy. These bacteria express genes encoding for extended-spectrum β-lactamases (ESBL or bear a plasmid-mediated AmpC that induce phenotypically a resistance to the last-generation cephalosporins; even more worrying is the rapid increase of Enterobacteriaceae carrying genes conferring resistance to carbapenems (CPE. Materials and methods. The gut may serve as reservoir for these antibiotic drug-resistant bacteria: as a consequence, the rapid detection of drug resistant Enterobacteriaceae from rectal swabs is an important tool to identify rectal carriage of resistant bacteria. This procedure is the basic tool to successfully implement the infection control measures in the hospital wards. The study evaluated the capability of the HB&L ESBL/AmpC and CARBAPENEMASE screening kit (Alifax, Padua, Italy to rapidly identify the drug resistant enterobacteriaceae from rectal swabs: the performance was compared with the conventional method. Results and conclusions. The overall agreement was very good (91% for the detection of ESBL-AmpC, and 96.2% for the identification of CPE; this method is thus an efficient tool to quickly report positive multidrug resistant bacteria in rectal swabs.

  19. Mossambicus tilapia (Oreochromis mossambicus) collected from water bodies impacted by urban waste carries extended-spectrum beta-lactamases and integron-bearing gut bacteria.

    Science.gov (United States)

    Marathe, Nachiket P; Gaikwad, Swapnil S; Vaishampayan, Ankita A; Rasane, Mandar H; Shouche, Yogesh S; Gade, Wasudev N

    2016-09-01

    Oreochromis mossambicus (Peters 1852) (Tilapia) is one of the most consumed fish globally. Tilapia thrives well in environments 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, and studied the prevalence of resistance genes by molecular methods. A total of 91 bacterial strains were obtained, which include fish 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) blaCTX-M, blaSHV and aac(6')-Ib-cr gene was 38 percent, 24 percent, 38 percent, 31 percent and 31 percent respectively. Forty-two percent of the Enterobacteriaceae strains carried blaCTX-M gene, which is a common ESBL gene in clinics. The study demonstrates that tilapia found in the polluted waters can serve as reservoirs and an alternative route for human exposure to clinically important ARG-carrying bacteria. The consumption and handling of these fish may pose a potential health risk.

  20. Trends in Extended Spectrum Beta-Lactamase (ESBL) Producing Enterobacteriaceae and ESBL Genes in a Dutch Teaching Hospital, Measured in 5 Yearly Point Prevalence Surveys (2010-2014)

    NARCIS (Netherlands)

    Willemsen, Ina; Oome, Stijn; Verhulst, Carlo; Pettersson, Annika; Verduin, Kees; Kluytmans, Jan

    2015-01-01

    This paper describes the trends in prevalence of ESBL producing Enterobacteriaceae (ESBL-E) and ESBL genes, measured in five consecutive yearly Point Prevalence Surveys (PPS). All patients present in the hospital and in a day-care clinic (including patients on dialysis) on the day of the survey,

  1. 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...

  2. The Prevalence of Extended-Spectrum Beta-Lactamase-Producing Multidrug-Resistant Escherichia Coli in Poultry Chickens and Variation According to Farming Practices in Punjab, India

    Science.gov (United States)

    Mandal, Siddhartha; Hayer, Shivdeep; Sran, Mandeep; Zehra, Asima; Patel, Sunny J.; Kaur, Ravneet; Chatterjee, Leena; Mishra, Savita; Das, B.R.; Singh, Parminder; Singh, Randhir; Gill, J.P.S.

    2017-01-01

    Background: Agricultural use of antimicrobials in subtherapeutic concentrations is increasing in response to the rising demand for food animal products worldwide. In India, the use of antimicrobials in food animal production is unregulated. Research suggests that many clinically important antimicrobials are used indiscriminately. This is the largest study to date in India that surveys poultry production to test for antimicrobial resistance and the occurrence of extended-spectrum β-lactamases (ESBLs) modulated by farming and managerial practices. Objectives: Our goal was to survey poultry production for resistance to eleven clinically relevant antimicrobials and phenotypic occurrence of ESBLs as modulated by farming and managerial practices. Methods: Eighteen poultry farms from Punjab were surveyed, and 1,556 Escherichia coli isolates from 530 birds were tested for susceptibility to 11 antimicrobials using the disk diffusion method and validated using VITEK 2 (bioMérieux, Marcy-L’Étoile, France). Samples from 510 of these birds were phenotypically tested for ESBL production using the combination disk method and confirmed using VITEK 2. Generalized linear mixed models were used to infer differences in resistance profiles associated with different farming practices and facility types. Results: Resistance profiles were significantly different between broiler and layer farms. Broiler farms were 2.2 [ampicillin (AMP), p=0.017] to 23 [nalidixic acid (NX), pproducing strains (87% compared to 42% in layers), was observed in broiler farms. Conclusions: Our findings suggest that unregulated use of clinically relevant antimicrobials in Indian broiler and layer farms may contribute to the emergence of resistance and support the need to curb the nontherapeutic use of medically important antimicrobials in food animal production. https://doi.org/10.1289/EHP292 PMID:28749780

  3. Trends in Extended Spectrum Beta-Lactamase (ESBL Producing Enterobacteriaceae and ESBL Genes in a Dutch Teaching Hospital, Measured in 5 Yearly Point Prevalence Surveys (2010-2014.

    Directory of Open Access Journals (Sweden)

    Ina Willemsen

    Full Text Available This paper describes the trends in prevalence of ESBL producing Enterobacteriaceae (ESBL-E and ESBL genes, measured in five consecutive yearly Point Prevalence Surveys (PPS. All patients present in the hospital and in a day-care clinic (including patients on dialysis on the day of the survey, were screened for perianal ESBL-E carriage. Perianal swabs were taken and cultured using an enrichment broth and a selective agar plate. Both phenotypic and genotypic methods were used to detect the production of ESBL, presence of ESBL-genes and clonal relatedness. Out of 2,695 patients, 135 (5.0% were tested ESBL-E positive. The overall ESBL-E prevalence was stable over the years. Overall 5.2% of all ESBL-E were acquired by nosocomial transmission. A relative decrease of CTX-M-1-1-like ESBL genes (from 44 to 25%, p = 0.026 was observed, possibly related to the strong (>60% decrease in antibiotic use in livestock in our country during the same period.

  4. A Rapid Phenotypic Whole Cell Screening Approach for the Identification of Small Molecule Inhibitors that Counter Beta-lactamase Resistance in Pseudomonas aeruginosa

    Science.gov (United States)

    Collia, Deanna; Bannister, Thomas D.; Tan, Hao; Jin, Shouguang; Langaee, Taimour; Shumate, Justin; Scampavia, Louis; Spicer, Timothy P.

    2017-01-01

    Pseudomonas aeruginosa is an opportunistic human pathogen which is prevalent in hospitals and continues to develop resistance to multiple classes of antibiotics. Historically, β-lactam antibiotics have been the first line of therapeutic defense. However, the emergence of multidrug-resistant (MDR) strains of P. aeruginosa, such as AmpC β-lactamase overproducing mutants, limits the effectiveness of current antibiotics. Among AmpC hyper producing clinical isolates, inactivation of AmpG, which is essential for the expression of AmpC, increases bacterial sensitivity to β-lactam antibiotics. We hypothesize that inhibition of AmpG activity will enhance the efficacy of β-lactams against P. aeruginosa. Here, using a highly drug resistant AmpC inducible laboratory strain PAO1, we describe an ultra-high throughput whole cell turbidity assay designed to identify small molecule inhibitors of the AmpG. We screened 645K compounds to identify compounds with the ability to inhibit bacterial growth in the presence of Cefoxitin; an AmpC inducer, and identified 2,663 inhibitors which were also tested in the absence of Cefoxitin to determine AmpG specificity. The Z′ and S:B were robust at 0.87 ± 0.05 and 2.2 ± 0.2, respectively. Through a series of secondary and tertiary studies, including a novel luciferase based counterscreen, we ultimately identified 8 potential AmpG specific inhibitors. PMID:28850797

  5. Evaluation of a commercial microarray as a confirmation test for the presence of extended-spectrum beta-lactamases in isolates from the routine clinical setting.

    NARCIS (Netherlands)

    Platteel, T.N.; Stuart, J.W.; Voets, G.M.; Scharringa, J.; Sande, N. van de; Fluit, A.C.; Leverstein-van Hall, M.A.; Sturm, P.D.J.; et al.,

    2011-01-01

    Since the diagnostic characteristics of the Check-KPC ESBL microarray as a confirmation test on isolates obtained in a routine clinical setting have not been determined, we evaluated the microarray in a random selection of 346 clinical isolates with a positive ESBL screen test (MIC >1 mg/L for

  6. Epidemiology of Extended-Spectrum beta-Lactamase-Producing E-coli and Vancomycin-Resistant Enterococci in the Northern Dutch-German Cross-Border Region

    NARCIS (Netherlands)

    Zhou, Xuewei; Garcia-Cobos, Silvia; Ruijs, Gijs J. H. M.; Kampinga, Greetje A.; Arends, Jan P.; Borst, Dirk M.; Moller, Lieke V.; Holman, Nicole D.; Schuurs, Theo A.; van Coppenraet, Lesla E. Bruijnesteijn; Weel, Jan F.; van Zeijl, Jan H.; Koeck, Robin; Rossen, John W. A.; Friedrich, Alexander W.

    2017-01-01

    Objectives: To reveal the prevalence and epidemiology of extended-spectrum β-lactamase (ESBL)- and/or plasmid AmpC (pAmpC)- and carbapenemase (CP) producing Enterobacteriaceae and vancomycin-resistant enterococci (VRE) across the Northern Dutch-German border region. Methods: A point-prevalence study

  7. Characteristics of gram-negative urinary tract infections caused by extended spectrum beta lactamases: pivmecillinam as a treatment option within South Dublin, Ireland

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    Fardod O’Kelly

    2016-11-01

    Full Text Available Abstract Background The prevalence of urinary tract infections (UTIs caused by extended-spectrum β-lactamase (ESBL-producing Enterobacteriaceae is increasing and the therapeutic options are limited, especially in primary care. Recent indications have suggested pivmecillinam to be a suitable option. This pilot study aimed to assess the viability of pivmecillinam as a therapeutic option in a Dublin cohort of mixed community and healthcare origin. Methods A prospective measurement of mean and fractional inhibitory concentrations of antibiotic use in 95 patients diagnosed with UTI caused by ESBL-producing Enterobacteriaceae was carried out. 36 % patients were from general practice, 40 % were admitted to hospital within south Dublin, and 25 % samples arose from nursing homes. EUCAST breakpoints were used to determine if an isolate was sensitive or resistant to antibiotic agents. Results Sixty-nine percent of patients (N = 66 with urinary ESBL isolates were female. The mean age of females was 66 years compared with a mean age of 74 years for males. Thirty-six percent of isolates originated from primary care, hospital inpatients (26 %, and nursing homes (24 %. The vast majority of ESBL isolates were E. coli (80 %. The E tests for mecillinam and co-amoxiclav had concentration ranges from 0.16 mg/L up to 256 mg/L. The mean inhibitory concentration (MIC of mecillinam ranged from 0.25 to 256 mg/L, while co-amoxiclav MICs ranged from 6 to 256 mg/L. The percentage of isolates resistant to mecillinam and co-amoxiclav was found to be 5.26 and 94.74 % respectively. Conclusions This is the first study exploring the use of pivmecillinam in an Irish cohort and has demonstrated that its use in conjunction with or without co-amoxiclav is an appropriate and useful treatment for urinary tract infections caused by ESBL-producing organisms.

  8. In vitro activity and stability against novel beta-lactamases of investigational beta-lactams (cefepime, cefpirome, flomoxef, SCE2787 and piperacillin plus tazobactam) in comparison with established compounds (cefotaxime, latamoxef and piperacillin).

    Science.gov (United States)

    Bauernfeind, A; Schweighart, S; Eberlein, E; Jungwirth, R

    1991-01-01

    The therapeutic perspectives of flomoxef, SCE 2787, cefpirome, cefepime, latamoxef, cefotaxime and of piperacillin plus tazobactam were comparatively evaluated by their in vitro activity against 1119 clinical isolates of 83 bacterial species. Escherichia coli, Klebsiella spp. Enterobacter sakazakii, Proteus spp. and Shigella spp. were about equally susceptible to the cephalosporins (MIC90: 0.06 to 0.5 mg/l), while the MIC90 for piperacillin plus tazobactam was between 2 and 16 mg/l. Enterobacter cloacae, Enterobacter aerogenes and Serratia spp. were most susceptible to SCE 2787, cefpirome and cefepime (MIC90: 0.06 to 2 mg/l) followed by latamoxef, cefotaxime, flomoxef and piperacillin plus tazobactam. For Citrobacter spp., Providencia spp. and Yersinia enterocolitica MIC90 were between 0.06 and 0.5 mg/l. Flomoxef was between 2 to 4 log2 less active against these species but more active than piperacillin plus tazobactam (MIC90: 2 and 8 mg/l). Morganella morganii and Hafnia alvei were most susceptible to cefepime, cefpirome and latamoxef (MIC90: 0.13 to 0.5 mg/l) while cefotaxime (MIC90: 8 mg/l) and piperacillin plus tazobactam (MIC90: 8 and greater than 64 mg/l) were the least active compounds. SCE 2787, cefepime and cefpirome were the most potent beta-lactams against the majority of the 13 species of non-fermentative bacilli (NFB) investigated (MIC90: 0.5 to 16 mg/l). The oxacephems were the least active compounds against NFB. Cefepime was the most active of the compounds included against Pseudomonas aeruginosa (MIC90: 16 mg/l). Haemophilus spp., Neisseria gonorrhoeae and Bordetella pertussis were most susceptible to cefotaxime (MIC90: 0.03 to 0.06 mg/l). Latamoxef had the lowest activity of all compounds against gram-positive cocci. Flomoxef was the most active compound against penicillinase producing Staphylococcus aureus and about equally active as the other betalactams against methicillin susceptible staphylococci of other staphylococcal species.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Spread of ISCR1 elements containing blaDHA-₁ and multiple antimicrobial resistance genes leading to increase of flomoxef resistance in extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Lee, Chen-Hsiang; Liu, Jien-Wei; Li, Chia-Chin; Chien, Chun-Chih; Tang, Ya-Fen; Su, Lin-Hui

    2011-09-01

    Increasing resistance to quinolones, aminoglycosides, and/or cephamycins in extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae exacerbates the already limited antibiotic treatment options for infections due to these microbes. In this study, the presence of resistance determinants for these antimicrobial agents was examined by PCR among ESBL-producing Klebsiella pneumoniae (ESBL-KP) isolates that caused bacteremia. Pulsed-field gel electrophoresis was used to differentiate the clonal relationship among the isolates studied. Transferability and the location of the resistance genes were analyzed by conjugation experiments, followed by DNA-DNA hybridization. Among the 94 ESBL-KP isolates studied, 20 isolates of flomoxef-resistant ESBL-KP were identified. They all carried a DHA-1 gene and were genetically diverse. CTX-M genes were found in 18 of the isolates. Among these DHA-1/CTX-M-producing K. pneumoniae isolates, ISCR1 was detected in 13 (72%) isolates, qnr genes (1 qnrA and 17 qnrB genes) were detected in 18 (100%), aac(6')-Ib-cr was detected in 11 (61%), and 16S rRNA methylase (all armA genes) was detected in 14 (78%). Four transconjugants were available for further analysis, and qnrB4, aac(6')-Ib-cr, armA, and bla(DHA-1) were all identified on these self-transferable bla(CTX-M)-carrying plasmids. The genetic environments of ISCR1 associated with armA, bla(DHA-1), and qnrB4 genes in the four transconjugants were identical. Replicon-type analysis revealed a FIIA plasmid among the four self-transferable plasmids, although the other three were nontypeable. The cotransfer of multiple resistance genes with the ISCR1 element-carrying plasmids has a clinical impact and warrants close monitoring and further study.

  10. 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

    2016-01-01

    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

  11. Detection of CTX-M-15 beta-lactamases in Enterobacteriaceae causing hospital- and community-acquired urinary tract infections as early as 2004, in Dar es Salaam, Tanzania.

    Science.gov (United States)

    Manyahi, Joel; Moyo, Sabrina J; Tellevik, Marit Gjerde; Ndugulile, Faustine; Urassa, Willy; Blomberg, Bjørn; Langeland, Nina

    2017-04-17

    The spread of Extended Spectrum β-lactamases (ESBLs) among Enterobacteriaceae and other Gram-Negative pathogens in the community and hospitals represents a major challenge to combat infections. We conducted a study to assess the prevalence and genetic makeup of ESBL-type resistance in bacterial isolates causing community- and hospital-acquired urinary tract infections. A total of 172 isolates of Enterobacteriaceae were collected in Dar es Salaam, Tanzania, from patients who met criteria of community and hospital-acquired urinary tract infections. We used E-test ESBL strips to test for ESBL-phenotype and PCR and sequencing for detection of ESBL genes. Overall 23.8% (41/172) of all isolates were ESBL-producers. ESBL-producers were more frequently isolated from hospital-acquired infections (32%, 27/84 than from community-acquired infections (16%, 14/88, p Enterobacteriaceae causing both hospital- and community-acquired infections in Tanzania.

  12. Evaluation of meat, fruit and vegetables from retail stores in five United Kingdom regions as sources of extended-spectrum beta-lactamase (ESBL)-producing and carbapenem-resistant Escherichia coli.

    Science.gov (United States)

    Randall, L P; Lodge, M P; Elviss, N C; Lemma, F L; Hopkins, K L; Teale, C J; Woodford, N

    2017-01-16

    We determined the prevalence and types of extended-spectrum β-lactamase (ESBL)-producing and carbapenem-resistant Escherichia coli in raw retail beef, chicken, pork, fruit and vegetables in five UK regions in 2013-14. Raw meat (n=397), and fruit and vegetable samples (n=400) were purchased from retail stores in London, East Anglia, North West England, Scotland and Wales. Samples were tested for the presence of ESBL-producing E. coli by plating enriched samples on CHROMagar CTX and CHROMagar ESBL, for AmpC-type E. coli by plating on "CHROMagar FOX" (CHROMagar ECC+16mg/L cefoxitin), and for carbapenem-resistant E. coli by plating on CHROMagar KPC. Additionally, pre-enrichment counts were performed on the above agars, and on CHROMagar ECC. Isolates of interest were characterised by MALDI-ToF to confirm identification, by PCR for bla CIT, bla CTX-M, bla OXA , bla SHV and bla TEM genes; ESBL or bla CIT genes were sequenced. Only 1.9% and 2.5% of beef and pork samples, respectively were positive for ESBL-producing E. coli after enrichment compared with 65.4% of chicken samples. 85.6% positive samples from chicken meat carried bla CTX-M-1 ; bla CTX-M-15 was not detected. None of the fruits or vegetables yielded ESBL-producing E. coli and none of the meat, fruit or vegetable samples yielded carbapenem-resistant E. coli. Retail chicken was more frequently a source of ESBL-producing E. coli than were beef, pork, fruit or vegetables. None of the foodstuffs yielded E. coli with CTX-M-15 ESBL, which dominates in human clinical isolates in the UK, and none yielded carbapenem-resistant E. coli. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  13. Multiple-locus variable number of tandem repeats (VNTR) fingerprinting (MLVF) and antibacterial resistance profiles of extended spectrum beta lactamase (ESBL) producing Pseudomonas aeruginosa among burnt patients in Tehran.

    Science.gov (United States)

    Jabalameli, Fereshteh; Mirsalehian, Akbar; Sotoudeh, Nazli; Jabalameli, Leila; Aligholi, Marzieh; Khoramian, Babak; Taherikalani, Morovat; Emaneini, Mohammad

    2011-11-01

    Extended spectrum β-lactamase (ESBL)-producing trait was present in 48 out of the 112 (42.8%) Pseudomonas aeruginosa isolates collected from burn wound infections during a 12-month period. The presence of oxa-10, per-1, veb-1 and ges genes and the multiple-locus variable number of tandem repeats (VNTR) fingerprinting (MLVF) of 112 P. aeruginosa strains were determined by PCR and multiplex PCR. Disk diffusion methods were used to determine the susceptibility of the isolates to antimicrobial agents as instructed by CLSI. All ESBL isolates were resistant to aztreonam, cefepime, cefotaxime, cefpodoxime, ceftazidime, ceftriaxone and ofloxacin. Fewer than 60% of ESBL isolates were resistant to imipenem, meropenem, and piperacillin-tazobactam but more than 90% were resistant to amikacin, ciprofloxacin, levofloxacin, ticarcillin and tobramycin. The most prevalent ESBL genes included oxa-10 (70%) and per-1 (50%) followed by veb-1 (31.3%). The gene encodes GES enzyme did not detect in any isolates. A total of 100 P. aeruginosa strains were typed by MLVF typing method. MLVF produced 42 different DNA banding patterns. These data indicate that different MLVF types infect burn wounds in patients at a hospital in Tehran and also suggest an alarming rate of ESBL-producing isolates in this test location. Copyright © 2011 Elsevier Ltd and ISBI. All rights reserved.

  14. 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

    and commensal E. coli isolates collected from 2006 to 2012 from humans, retail poultry meat, broilers, and dogs. Multilocus sequence typing (MLST), antimicrobial susceptibility testing, and conjugation were performed in conjunction with plasmid replicon typing, plasmid multilocus sequence typing (p......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...... were conjugative and did not (96%) cotransfer resistance to antimicrobials other than cephalosporins. The main replicon types identified were IncI1-I gamma (55%) and IncK (39%). Isolates from different host species mainly carried distinct plasmid subtypes. Seven of the 18 human isolates harbored IncI1...

  15. 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

    2014-01-01

    -generation cephalosporin use and 19 herds with previous frequent use were included. The ESBL-producing isolates detected in humans and pigs were characterized by ESBL genotype, PFGE, susceptibility to non-b-lactam antibiotics and phylotype, and selected isolates were characterized by multilocus sequence typing (MLST......Objectives: To compare and characterize extended-spectrum b-lactamase (ESBL)-producing Escherichia coli from pigsties, pig farmers and their families on farms with previous high or no use of third- or fourth-generation cephalosporins. Methods: Twenty farms with no third- or fourth...

  16. Antimicrobial multiple resistance index, minimum inhibitory concentrations, and extended-spectrum beta-lactamase producers of Proteus mirabilis and Proteus vulgaris strains isolated from domestic animals with various clinical manifestations of infection

    Directory of Open Access Journals (Sweden)

    Vanessa Zappa

    2017-05-01

    Full Text Available Proteus spp. are opportunistic multidrug resistant enterobacteria associated with diverse clinical diseases in domestic animals. However, Proteus infections in domestic animals are often misdiagnosed or considered contaminants in microbiological cultures rather than a primary agent of disease. Descriptions of Proteus infections in domestic animals are typically restricted to case reports, retrospective studies, or surveillance of other microorganisms. The present study investigated multiple antibiotic resistance indices, minimum inhibitory concentrations (MICs, and ESBL production in 73 strains of Proteus mirabilis (n = 69 and Proteus vulgaris (n = 4 isolated from domestic animals with various clinical manifestations. In dogs, the pathogen was most commonly associated with cystitis (48.21, enteritis (21.42%, otitis (14.29%, and conjunctivitis (3.57%. In bovines, the microorganism was predominant in cases of enteritis (22.22%, abscess (11.11%, otitis (11.11%, omphalitis (11.11%, and peritonitis (11.11%, and in organ fragments (11.11%. In equines (50.0% and cats (100.0%, diarrhea was the main clinical sign. In vitro standard disk diffusion assay showed that the most effective antimicrobials against the isolates were imipenem (98.63, norfloxacin (95.89, amikacin (95.89, levofloxacin (90.41, ceftriaxone (87.64, and florfenicol (87.67. In contrast, the isolates commonly showed resistance to novobiocin (95.89, azithromycin (57.53, and trimethropim/sulfamethoxazole (39.73. Among the 73 isolates, the efficacy of amoxicillin/clavulanic acid, gentamicin, ceftriaxone, and ciprofloxacin according to MICs was 87.67%, 86.30%, 84.93%, and 82.19%, respectively. The MIC50 values of amoxicillin/clavulanic acid, ceftriaxone, ciprofloxacin, and gentamicin were, respectively, 1.0, 0.004, 0.03, and 1.0 µg/mL. Thirty-three strains (45.21% showed a antimicrobial multiple resistance index of ? 0.3. Multidrug resistance profiles of isolates were observed most frequently in dogs (n = 25; 75.76%, particularly in those with cystitis (n = 13; 52.0%, followed by bovines (n = 4; 12.12%, equines (n = 2; 6.06%, and cats (n = 2; 6.06%. Two (2.7% strains, obtained from canine skin and feces, were diagnosed phenotypically as ESBL-producers. Here, we observed the diversity of Proteus infections in domestic animals. The presence of multidrug-resistant isolates and ESBL-producers reinforces the need for appropriate antimicrobial use and in vitro antimicrobial tests to support therapy.

  17. Antibiotic resistance profile and phenotypic detection of beta ...

    African Journals Online (AJOL)

    Abstract. Background: Cockroaches are carriers of numerous microorganisms. However, there is paucity of information on their role as potential reservoir for beta-lactamase producers. Objectives: This research determined the antibiotics susceptibility profile of Beta-lactamase producing Gram-negative bacteria isolated from ...

  18. 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 metal...

  19. 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...

  20. Presence of ESBL/AmpC-producing Escherichia coli in the broiler production pyramid: a descriptive study.

    NARCIS (Netherlands)

    Dierikx, C.M.; Goot, van der 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

  1. A Novel IncA/C1 Group Conjugative Plasmid, Encoding VIM-1 Metallo-Beta-Lactamase, Mediates the Acquisition of Carbapenem Resistance in ST104 Klebsiella pneumoniae Isolates from Neonates in the Intensive Care Unit of V. Monaldi Hospital in Naples.

    Science.gov (United States)

    Esposito, Eliana P; Gaiarsa, Stefano; Del Franco, Mariateresa; Crivaro, Valeria; Bernardo, Mariano; Cuccurullo, Susanna; Pennino, Francesca; Triassi, Maria; Marone, Piero; Sassera, Davide; Zarrilli, Raffaele

    2017-01-01

    The emergence of carbapenemase producing Enterobacteriaceae has raised major public health concern. The aim of this study was to investigate the molecular epidemiology and the mechanism of carbapenem resistance acquisition of multidrug-resistant Klebsiella pneumoniae isolates from 20 neonates in the neonatal intensive care unit (NICU) of the V. Monaldi Hospital in Naples, Italy, from April 2015 to March 2016. Genotype analysis by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) identified PFGE type A and subtypes A1 and A2 in 17, 2, and 1 isolates, respectively, and assigned all isolates to sequence type (ST) 104. K. pneumoniae isolates were resistant to all classes of β-lactams including carbapenems, fosfomycin, gentamicin, and trimethoprim-sulfamethoxazole, but susceptible to quinolones, amikacin, and colistin. Conjugation experiments demonstrated that resistance to third-generation cephems and imipenem could be transferred along with an IncA/C plasmid containing the extended spectrum β-lactamase bla SHV -12 and carbapenem-hydrolyzing metallo-β-lactamase bla V IM-1 genes. The plasmid that we called pIncAC_KP4898 was 156,252 bp in size and included a typical IncA/C backbone, which was assigned to ST12 and core genome (cg) ST12.1 using the IncA/C plasmid MLST (PMLST) scheme. pIncAC_KP4898 showed a mosaic structure with bla V IM-1 into a class I integron, bla SHV -12 flanked by IS6 elements, a mercury resistance and a macrolide 2'-phosphotransferase clusters, ant(3″), aph(3″), aacA4, qnrA1, sul1 , and dfrA14 conferring resistance to aminoglycosides, quinolones, sulfonamides, and trimethoprim, respectively, several genes predicted to encode transfer functions and proteins involved in DNA transposition. The acquisition of pIncAC_KP4898 carrying bla V IM-1 and bla SHV -12 contributed to the spread of ST104 K. pneumoniae in the NICU of V. Monaldi Hospital in Naples.

  2. A Novel IncA/C1 Group Conjugative Plasmid, Encoding VIM-1 Metallo-Beta-Lactamase, Mediates the Acquisition of Carbapenem Resistance in ST104 Klebsiella pneumoniae Isolates from Neonates in the Intensive Care Unit of V. Monaldi Hospital in Naples

    Directory of Open Access Journals (Sweden)

    Eliana P. Esposito

    2017-11-01

    Full Text Available The emergence of carbapenemase producing Enterobacteriaceae has raised major public health concern. The aim of this study was to investigate the molecular epidemiology and the mechanism of carbapenem resistance acquisition of multidrug-resistant Klebsiella pneumoniae isolates from 20 neonates in the neonatal intensive care unit (NICU of the V. Monaldi Hospital in Naples, Italy, from April 2015 to March 2016. Genotype analysis by pulsed-field gel electrophoresis (PFGE and multi-locus sequence typing (MLST identified PFGE type A and subtypes A1 and A2 in 17, 2, and 1 isolates, respectively, and assigned all isolates to sequence type (ST 104. K. pneumoniae isolates were resistant to all classes of β-lactams including carbapenems, fosfomycin, gentamicin, and trimethoprim–sulfamethoxazole, but susceptible to quinolones, amikacin, and colistin. Conjugation experiments demonstrated that resistance to third-generation cephems and imipenem could be transferred along with an IncA/C plasmid containing the extended spectrum β-lactamase blaSHV -12 and carbapenem-hydrolyzing metallo-β-lactamase blaV IM-1 genes. The plasmid that we called pIncAC_KP4898 was 156,252 bp in size and included a typical IncA/C backbone, which was assigned to ST12 and core genome (cg ST12.1 using the IncA/C plasmid MLST (PMLST scheme. pIncAC_KP4898 showed a mosaic structure with blaV IM-1 into a class I integron, blaSHV -12 flanked by IS6 elements, a mercury resistance and a macrolide 2′-phosphotransferase clusters, ant(3″, aph(3″, aacA4, qnrA1, sul1, and dfrA14 conferring resistance to aminoglycosides, quinolones, sulfonamides, and trimethoprim, respectively, several genes predicted to encode transfer functions and proteins involved in DNA transposition. The acquisition of pIncAC_KP4898 carrying blaV IM-1 and blaSHV -12 contributed to the spread of ST104 K. pneumoniae in the NICU of V. Monaldi Hospital in Naples.

  3. Índice de resistência múltipla aos antimicrobianos, concentração inibitória e beta-lactamases de espectro estendido em linhagens de Proteus mirabilis e Proteus vulgaris isoladas de diferentes afecções em animais domésticos

    OpenAIRE

    Zappa, Vanessa [UNESP

    2015-01-01

    In the last decades have been highlighted the increase number of infections in domestic animals and humans caused by opportunistic multidrug resistant enterobacteria, commonly associated to improper use of antimicrobials, including by Proteus species. However, Proteus infections in domestic animals have been misdiagnosed or the microorganism is considered a contaminant of microbiological cultures, besides to be a primary agent of diseases. Descriptions of Proteus infections in domestic animal...

  4. A novel chimeric peptide with antimicrobial activity.

    Science.gov (United States)

    Alaybeyoglu, Begum; Akbulut, Berna Sariyar; Ozkirimli, Elif

    2015-04-01

    Beta-lactamase-mediated bacterial drug resistance exacerbates the prognosis of infectious diseases, which are sometimes treated with co-administration of beta-lactam type antibiotics and beta-lactamase inhibitors. Antimicrobial peptides are promising broad-spectrum alternatives to conventional antibiotics in this era of evolving bacterial resistance. Peptides based on the Ala46-Tyr51 beta-hairpin loop of beta-lactamase inhibitory protein (BLIP) have been previously shown to inhibit beta-lactamase. Here, our goal was to modify this peptide for improved beta-lactamase inhibition and cellular uptake. Motivated by the cell-penetrating pVEC sequence, which includes a hydrophobic stretch at its N-terminus, our approach involved the addition of LLIIL residues to the inhibitory peptide N-terminus to facilitate uptake. Activity measurements of the peptide based on the 45-53 loop of BLIP for enhanced inhibition verified that the peptide was a competitive beta-lactamase inhibitor with a K(i) value of 58 μM. Incubation of beta-lactam-resistant cells with peptide decreased the number of viable cells, while it had no effect on beta-lactamase-free cells, indicating that this peptide had antimicrobial activity via beta-lactamase inhibition. To elucidate the molecular mechanism by which this peptide moves across the membrane, steered molecular dynamics simulations were carried out. We propose that addition of hydrophobic residues to the N-terminus of the peptide affords a promising strategy in the design of novel antimicrobial peptides not only against beta-lactamase but also for other intracellular targets. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd.

  5. Plasmid-Mediated Resistance in Enterobacteriaceae Changing Landscape and Implications for Therapy

    NARCIS (Netherlands)

    Schultsz, Constance; Geerlings, Suzanne

    2012-01-01

    Antimicrobial resistance is increasing worldwide, and pathogenic microorganism's that are resistant to all available antimicrobial agents are increasingly reported. Emerging plasmid-encoded extended-spectrum beta-lactamases (ESBLs) and carbapenemases are increasingly reported worldwide.

  6. 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...

  7. The evolutionary landscape of antifolate resistance in Plasmodium ...

    Indian Academy of Sciences (India)

    2011-08-19

    Aug 19, 2011 ... Studies of beta-lactamases in bacteria have shown that only a limited number of pathways to antibiotic resistance are accessible to evolution .... a glimpse into processes governing the much more complex phenomenon that is ...

  8. Evaluation of restoration of sensitivities of resistant staphylococcus aureus isolates by using cefuroxime and clavulanic acid in combination

    International Nuclear Information System (INIS)

    Jalil, A.; Din, I.; Khan, S.U.

    2008-01-01

    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)

  9. 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

    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....... 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...

  10. Seagulls of the Berlengas Natural Reserve of Portugal as Carriers of Fecal Escherichia coli Harboring CTX-M and TEM Extended-Spectrum Beta-Lactamases▿

    Science.gov (United States)

    Poeta, Patricia; Radhouani, Hajer; Igrejas, Gilberto; Gonçalves, Alexandre; Carvalho, Carlos; Rodrigues, Jorge; Vinué, Laura; Somalo, Sergio; Torres, Carmen

    2008-01-01

    Escherichia coli isolates containing the following extended-spectrum beta-lactamases have been detected in 11 of 57 fecal samples (19.3%) in Berlengas Island seagulls: TEM-52 (eight isolates), CTX-M-1 (one isolate), CTX-M-14a (one isolate), and CTX-M-32 (one isolate). Most of the extended-spectrum beta-lactamase-positive isolates harbored class 1 or class 2 integrons, which included different antibiotic resistance gene cassettes. PMID:18835997

  11. Incidence of temonera, sulphuhydryl variables and cefotaximase genes associated with ?-lactamase producing escherichia coli in clinical isolates

    OpenAIRE

    Isaiah, Ibeh Nnana; Nche, Bikwe Thomas; Nwagu, Ibeh Georgina; Nwagu, Ibeh Isaiah

    2011-01-01

    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 p...

  12. Antibacterial activity of the Antarctic bacterium Janthinobacterium sp. SMN 33.6 against multi-resistant Gram-negative bacteria

    Directory of Open Access Journals (Sweden)

    Geraldine Asencio

    2014-01-01

    Conclusions: The ethanolic extract of Janthinobacterium sp. SMN 33.6 possesses antibacterial activity against a chromosomal AmpC beta-lactamase-producing strain of Serratia marcescens, an extended-spectrum beta-lactamase-producing Escherichia coli and also against carbapenemase-producing strains of Acinetobacter baumannii and Pseudomonas aeruginosa. This becomes a potential and interesting biotechnological tool for the control of bacteria with multi-resistance to commonly used antibiotics.

  13. ORF Alignment: NC_003384 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available [Zymomonas mobilis] gb|AAQ67239.1| ... Bla [Transposon delivery vector pUT-miniTn5-gfp-tet] ... ... ... 3.5.2.6) precursor - Escherichia coli plasmids ... gb|AAC79082.1| beta lactamase [Transposon delivery...AG47772.1| beta-lactamase ... TEM-1 [Klebsiella pneumoniae] gb|AAG37887.1| ampicillin ... resistance [Tn10 delivery... vector pHV1249] gb|AAG37882.1| ... ampicillin resistance [Tn10 delivery

  14. Incidence of temonera, sulphuhydryl variables and cefotaximase genes associated with β-lactamase producing escherichia coli in clinical isolates

    Science.gov (United States)

    Isaiah, Ibeh Nnana; Nche, Bikwe Thomas; Nwagu, Ibeh Georgina; Nwagu, Ibeh Isaiah

    2011-01-01

    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 (PCT)phenotypic 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. PMID:22363078

  15. The role of outer membrane in Serratia marcescens intrinsic resistance to antibiotics.

    Science.gov (United States)

    Sánchez, L; Ruiz, N; Leranoz, S; Viñas, M; Puig, M

    1997-09-01

    Three different porins from Serratia marcescens were described. They were named Omp1, Omp2 and Omp3 and their molecular weights were 42, 40 and 39 kDa respectively. Omp2 and Omp3 showed osmoregulation and thermoregulation in a similar way to OmpC and OmpF of Escherichia coli. Permeability coefficients of the outer membrane of this species were calculated following the Zimmermann and Rosselet method. P values were similar to those obtained in Escherichia coli, which suggests that the chromosomal beta-lactamase would play a major role in the resistance of Serratia marcescens to beta-lactam antibiotics. Both MIC values and permeabilities were modified by salycilates and acetylsalycilate. Synergism between the outer membrane and the beta-lactamase was also evaluated. When bacteria grew in the presence of a beta-lactam in the medium, the beta-lactamase accounted for most of the resistance.

  16. Epidemiology of infections caused by multiresistant gram-negatives: ESBLs, MBLs, panresistant strains.

    Science.gov (United States)

    Rossolini, Gian Maria; Mantengoli, Elisabetta; Docquier, Jean-Denis; Musmanno, Rosa Anna; Coratza, Grazietta

    2007-07-01

    Microbial drug resistance is a growing problem of global magnitude. In gram-negative pathogens, the most important resistance problems are encountered in Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter, with increasing trends observed for all major anti-gram-negative agents (beta-lactams, fluoroquinolones and aminoglycosides). A matter of major concern is the emergence of new beta-lactamases capable of degrading the expanded-spectrum cephalosporins and/or carbapenems, such as the extended-spectrum beta-lactamases (ESBLs) and the carbapenemases. These beta-lactamase genes are often associated with resistance determinants to non-beta-lactam agents (e.g. aminoglycosides and fluoroquinolones), and strains producing ESBLs or carbapenemases often exhibit complex multidrug resistant phenotypes and sometimes are panresistant. The problem is worsened by the dearth of new agents active on multidrug-resistant Gram-negatives in the pipeline. The importance to develop better strategies to control resistance is underscored.

  17. Treatment for infections complications of experimental acute radiation sickness with sulacillin, a combined antibiotic

    International Nuclear Information System (INIS)

    Chertkov, K.S.; Revskaya, E.A.

    1999-01-01

    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 LD 90/45 [ru

  18. 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.

  19. Development of antibiotic resistance in Pseudomonas aeruginosa during two decades of antipseudomonal treatment at the Danish CF Center

    DEFF Research Database (Denmark)

    Ciofu, O; Giwercman, B; Pedersen, S S

    1994-01-01

    resistance in P. aeruginosa strains isolated from Danish CF patients over a period of 18 years by testing the in vitro efficacy of carbenicillin, piperacillin, ceftazidime, tobramycin and ciprofloxacin against P. aeruginosa strains collected in 1973 (51 strains), 1980 (80 strains), 1985 (58 strains...... was found between the MIC and the number of antipseudomonal courses of antibiotics. The proportion of resistant in vivo selected P. aeruginosa strains, presumed to be stably derepressed producers of chromosomal beta-lactamase, also increased significantly during the period studied. Our results confirm...... that the beta-lactamase production is an important mechanism of antibiotic resistance in P. aeruginosa....

  20. Udviling i anvendelse af antibiotika i dansk fødevareproduktion

    DEFF Research Database (Denmark)

    Jensen, Vibeke Frøkjær

    2011-01-01

    of 2010 was likely due to the announcement of the "yellow card" regulation. From July 2010, a voluntary two years stop of cephalosporins use in pigs was realized, due to increasing occurrence of extended spectrum beta lactamase (ESBL) resistance in animal and meat isolates; highest levels of ESBL...

  1. Enzyme activities and antibiotic susceptibility of colonial variants of Bacillus subtilis and Bacillus licheniformis.

    OpenAIRE

    Carlisle, G E; Falkinham, J O

    1989-01-01

    A nonmucoid colonial variant of a mucoid Bacillus subtilis strain produced less amylase activity and a transparent colonial variant of a B. licheniformis strain produced less protease activity compared with their parents. Antibiotic susceptibility patterns of the colonial variants differed, and increased resistance to beta-lactam antibiotics was correlated with increased production of extracellular beta-lactamase.

  2. Epidemiology of antimicrobial resistance and the effect of interventions in food-producing animals

    NARCIS (Netherlands)

    Dorado Garcia, A.|info:eu-repo/dai/nl/372621023

    2016-01-01

    This thesis explores the animal-human interface of the emerging antimicrobial resistance (AMR) problem. It focuses on two relevant bacterial species imposing a burden for human health: methicillin-resistant Staphylococcus aureus (MRSA) and (extended-spectrum beta-lactamase (ESBL)/AmpC-producing)

  3. Pharmacodynamics of Cefepime Combined with Tazobactam against Clinically Relevant Enterobacteriaceae in a Neutropenic Mouse Thigh Model

    NARCIS (Netherlands)

    Melchers, M.J.; Mil, A.C van; Lagarde, C.M.; Hartigh, J. den; Mouton, J.W.

    2017-01-01

    The lack of new antibiotics has prompted investigation of the combination of two existing agents-cefepime, a broad-spectrum cephalosporin, and tazobactam-to broaden their efficacy against extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae We determined the pharmacokinetic (PK) and

  4. Antimicrobial susceptibility, risk factors and prevalence of bla cefotaximase, temoneira, and sulfhydryl variable genes among Escherichia coli in community-acquired pediatric urinary tract infection

    Directory of Open Access Journals (Sweden)

    Kallyadan V Nisha

    2017-01-01

    Conclusion: ESBL-producing E. coli from community-acquired pediatric UTI carries more than one type of beta-lactamase coding genes correlating their increased antibiotic resistance. Aggressive infection control policy, routine screening for detecting ESBL isolates in clinical samples, and antimicrobial stewardship are the keys to prevent their dissemination in community settings.

  5. Molecular Typing of Enterobacteriaceae from Pig Holdings in North-Western Germany Reveals Extended- Spectrum and AmpC β-Lactamases Producing but no Carbapenem Resistant Ones

    NARCIS (Netherlands)

    Garcia-Cobos, Silvia; Koeck, Robin; Mellmann, Alexander; Frenzel, Julia; Friedrich, Alexander W.; Rossen, John W. A.

    2015-01-01

    The increase of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) in humans and in food-producing animals is of public health concern. The latter could contribute to spreading of these bacteria or their resistance genes to humans. Several studies have reported the isolation of

  6. Seagulls and beaches as reservoirs for multidrug-resistant Escherichia coli.

    Science.gov (United States)

    Simões, Roméo Rocha; Poirel, Laurent; Da Costa, Paulo Martins; Nordmann, Patrice

    2010-01-01

    A variety of extended-spectrum Beta-lactamase-producing Escherichia coli isolates, with a high rate of cefotaximase-15 resistance, were identified in seagull feces from Porto, Portugal, beaches. Beaches may therefore present a risk to public health because of the potential pathogen-spreading capacity of migratory birds.

  7. Drug: D08350 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available D08350 Drug Pheneticillin (INN); Phenethicillin; PEPC ... C17H20N2O5S D08350.gif ... A...8 ... beta-Lactamase sensitive penicillin ATC code: J01CE05 Chemical group: DG00538 ... Phenethicillin is called

  8. Longitudinal study of extended-spectrum-β-lactamase- and AmpC-Producing Enterobacteriaceae in household dogs

    NARCIS (Netherlands)

    Baede, V.O.; Wagenaar, J.A.; Broens, E.M.; Duim, Birgitta; Dohmen, Wietske; Nijsse, Rolf; Timmerman, Arjen J.; Hordijk, Joost

    2015-01-01

    A longitudinal study was performed to (i) investigate the continuity of shedding of extended-spectrum-beta-lactamase (ESBL)-producing Enterobacteriaceae in dogs without clinical signs, (ii) identify dominant plasmid-mediated ESBL genes, and (iii) quantify ESBL-producing Enterobacteriaceae in

  9. Liamocins, sophorolipids, and frost grape polysaccharides. New carbohydrate research from the USDA’s NCAUR national laboratory

    Science.gov (United States)

    The USDA’s National Center for Agricultural Utilization Research has a prominent history in carbohydrate research, including the development of xanthan gum, ‘super slurper’ polysaccharides, beta-dextrans, alternan, and beta lactamase antibiotics (penicillins), as well as analytical tools such as ald...

  10. Emergence of New Delhi metalo-β- lactamase (NDM-1) – producing ...

    African Journals Online (AJOL)

    The New Delhi metallo-beta- lactamase (NDM-1) gene is an emerging well acknowledged public health threats among human and animal pathogens worldwide. Since its first ... The Gram negative bacteria were. identified by conventional bacteriological procedures and with16s RNA PCR method. In all, 7.3% (4/55) of the ...

  11. Selection and persistence of CTX-M-producing Escherichia coli in the intestinal flora of pigs treated with amoxicillin, ceftiofur, or cefquinome

    DEFF Research Database (Denmark)

    Cavaco, Lina; Abatih, E.; Aarestrup, Frank Møller

    2008-01-01

    Extended-spectrum beta-lactamases (ESBLs), mainly of the CTX-M family, have been associated with Escherichia coli strains of animal origin in Europe. An in vivo experiment was performed to study the effects of veterinary beta-lactam drugs on the selection and persistence of ESBL-producing E. coli...

  12. Multidrug resistant bacteria in companion animals: impact on animal health and zoonotic aspects

    DEFF Research Database (Denmark)

    Damborg, Peter Panduro

    -resistant bacteria include methicillin-resistant Staphylococcus pseudintermedius (MRSP), methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae. These bacteria will be described with focus on their prevalence across Europe, their impact on animal...

  13. Fulltext PDF

    Indian Academy of Sciences (India)

    IAS Admin

    and Diverse Implications in the Natural World. Science Academies' Summer Research Fellowship. Programme for Students and Teachers – 2016. Classics. On the Spontaneous. Fission of Uranium. Shyamadas Chatterjee. 856. 860. 861. 816 Split Beta-Lactamase Complementation Assay. A Search for the Molecular Better ...

  14. Outbreak of NDM-1-Producing Klebsiella pneumoniae in a Dutch Hospital, with Interspecies Transfer of the Resistance Plasmid and Unexpected Occurrence in Unrelated Health Care Centers

    NARCIS (Netherlands)

    Bosch, Thijs; Lutgens, Suzanne P M; Hermans, Mirjam H A; Wever, Peter C; Schneeberger, Peter M.; Renders, Nicole H M; Leenders, Alexander C. A. P.; Kluytmans, Jan A. J. W.; Schoffelen, Annelot F; Notermans, Daan; Witteveen, Sandra; Bathoorn, Erik; Schouls, Leo M.

    In the Netherlands, the number of cases of infection with New Delhi metallo-beta-lactamase (NDM)-positive Enterobacteriaceae is low. Here, we report an outbreak of NDM-1-producing Klebsiella pneumoniae infection in a Dutch hospital with interspecies transfer of the resistance plasmid and unexpected

  15. Outbreak of NDM-1-Producing Klebsiella pneumoniae in a Dutch Hospital, with Interspecies Transfer of the Resistance Plasmid and Unexpected Occurrence in Unrelated Health Care Centers.

    NARCIS (Netherlands)

    Bosch, Thijs; Lutgens, Suzanne P M; Hermans, Mirjam H A; Wever, Peter C; Schneeberger, Peter M; Renders, Nicole H M; Leenders, Alexander C A P; Kluytmans, Jan A J W; Schoffelen, Annelot; Notermans, Daan; Witteveen, Sandra; Bathoorn, Erik; Schouls, Leo M

    In the Netherlands, the number of cases of infection with New Delhi metallo-beta-lactamase (NDM)-positive Enterobacteriaceae is low. Here, we report an outbreak of NDM-1-producing Klebsiella pneumoniae infection in a Dutch hospital with interspecies transfer of the resistance plasmid and unexpected

  16. Khan et al., Afr J Tradit Complement Altern Med. (2015) 12(S):70-77 ...

    African Journals Online (AJOL)

    has become a significant problem and can be controlled with continued surveillance or monitoring for rationally improved antibiotics usage, in order to prevent the drug resistance and re-emergence of infection. Key words: Klebsiella pneumoniae, ESBL (Extended-spectrum beta-lactamase), API (Analytical Profile Index).

  17. Bacterial contamination of water samples in Gabon, 2013

    Directory of Open Access Journals (Sweden)

    Jonas Ehrhardt

    2017-10-01

    Full Text Available Contamination of water is a major burden in the public health setting of developing countries. We therefore assessed the quality of water samples in Gabon in 2013. The main findings were a contamination rate with coliforms of 13.5% and the detection of a possible environmental reservoir for extended spectrum beta-lactamase-producing bacteria.

  18. [Antibiotic resistance: measures urgently needed].

    NARCIS (Netherlands)

    Kluytmans, J.; Vandenbroucke-Grauls, C.M.; Meer, J.W.M. van der

    2010-01-01

    Antimicrobial resistance is increasing rapidly and there are hardly any new antimicrobial agents to be expected in the coming years. The number of patients affected by extended spectrum beta-lactamase producing organisms (ESBLs) is rising and there are strong indications that this is caused in part

  19. The prevalence of nosocomial infections caused by Enterobacter cloacae and antibiotic resistant patterns in samples isolated from patients in two hospitals in Tehran

    Directory of Open Access Journals (Sweden)

    Rahbar M

    2012-06-01

    Conclusion: This study clearly showed the high prevalence of resistance to broad-spectrum beta-lactam antibiotics in the isolated E. cloacae among which 5% were multi drug resistant. All the isolated E. cloacae were susceptible to Colistin. These results can be alarming for physicians treating resistant E. cloacae infections, especially extended-spectrum beta-lactamase producing species.

  20. Colonization of Enteroaggregative Escherichia coli and Shiga toxin-producing Escherichia coli in chickens and humans in southern Vietnam

    NARCIS (Netherlands)

    Trung, Nguyen Vinh; Nhung, Hoang Ngoc; Carrique-Mas, Juan J.; Mai, Ho Huynh; Tuyen, Ha Thanh; Campbell, James; Nhung, Nguyen Thi; van Minh, Pham; Wagenaar, Jaap A.; Mai, Nguyen Thi Nhu; Hieu, Thai Quoc; Schultsz, Constance; Hoa, Ngo Thi

    2016-01-01

    Enteroaggregative (EAEC) and Shiga-toxin producing Escherichia coli (STEC) are a major cause of diarrhea worldwide. E. coli carrying both virulence factors characteristic for EAEC and STEC and producing extended-spectrum beta-lactamase caused severe and protracted disease during an outbreak of E.

  1. Research Article

    African Journals Online (AJOL)

    2017-01-01

    Jan 1, 2017 ... Infection du tractus urinaire chez l'enfant: aspect épidémiologiqueset bactériologiques au centre hospitalier universitaire pédiatrique Charles de Gaule de Ouagadougou (Burkina Faso). 2009. [18] Ghuysen J.M. Molecular structures of penicillin-bending proteins and beta-lactamases.l. Trends Microbio.

  2. Distribution of mecA gene amongst Staphylococcus aureus isolates

    African Journals Online (AJOL)

    Dr Olaleye

    Antibiotics susceptibility testing including methicillin sensitivity testing, beta lactamase testing, PCR for detection of mecA gene, and minimum inhibitory concentrations to methicillin were carried out on all the 194 isolates of S. aureus. Among the 194 strains, 40 (20.6%) were MRSA using 10 µg methicillin disc. PCR analysis.

  3. lactamase in clinical isolates of Escherichia coli

    African Journals Online (AJOL)

    Jane

    2011-08-22

    Aug 22, 2011 ... The beta lactamase enzyme producing E. coli, resistant to β-lactam antibiotics, created many problems ... Key words: Escherichia coli, β-lactamase enzymes, TEM-type extended spectrum ... difficulties in treatment using antibiotics that are currently ... and chloramphenicol (30 µg) (Mast Diagnostics Ltd., UK).

  4. Khan et al., Afr J Tradit Complement Altern Med. (2015) 12(2):60-64 ...

    African Journals Online (AJOL)

    cadewumi

    3Faculty of Medicine and Allied Medical sciences, .... an average weight of 1.5 kg was purchased fromthe Animal market Hussain Agahi Multan, After ... Animals were divided into 4 groups, each group containing 3 rabbits as follows: ..... Inhibition of beta-lactamase enzyme of Pseudomonas aeruginosa by clavulanic acid.

  5. A Comparative Study on the Antimicrobial Susceptibility Patterns of ...

    African Journals Online (AJOL)

    ... Laboratory of Lagos University Teaching Hospital (LUTH). Isolates were obtained from urine and swabs of wounds, ear, throat and eye. Identification was carried out by conventional methods and antimicrobial susceptibility was investigated by disk diffusion method. All isolates were tested for beta-lactamase production.

  6. Prevalence of methicillin-resistant Staphylococcus aureus and ...

    African Journals Online (AJOL)

    Wound colonization by microorganisms is most frequently polymicrobial and incidences of high level resistance among bacterial isolates from wounds have been reported. Methicillin-resistant Staphylococcus aureus (MRSA) and extendedspectrum beta-lactamase (ESBL) producing Gram-negative bacteria both constitute ...

  7. Outbreak of Serratia marcescens colonization and infection traced to a Healthcare worker with long-term carriage on the hands

    NARCIS (Netherlands)

    de Vries, Jutte J. C.; Baas, Willy H.; van der Ploeg, Kees; Heesink, Albert; Degener, John E.; Arends, Jan P.

    2006-01-01

    objective. To reveal the source of a nosocomial outbreak of colonization and infection with a strain of Serratia marcescens positive for Guiana extended-spectrum beta-lactamase 1 (GES-1) that occurred among patients in a neurosurgical intensive care unit (ICU) in a Dutch university medical center

  8. Mechanisms of proton relay and product release by Class A β-lactamase at ultrahigh resolution

    Energy Technology Data Exchange (ETDEWEB)

    Lewandowski, Eric M. [Department of Molecular Medicine, University of South Florida College of Medicine, Tampa FL USA; Lethbridge, Kathryn G. [Department of Molecular Medicine, University of South Florida College of Medicine, Tampa FL USA; Sanishvili, Ruslan [GMCA@APS, X-ray Science Division, Advanced Photon Source, Argonne National Laboratory, IL USA; Skiba, Joanna [Department of Organic Chemistry, Faculty of Chemistry, University of Lodz, Poland; Kowalski, Konrad [Department of Organic Chemistry, Faculty of Chemistry, University of Lodz, Poland; Chen, Yu [Department of Molecular Medicine, University of South Florida College of Medicine, Tampa FL USA

    2017-11-20

    The beta-lactam antibiotics inhibit penicillin-binding proteins (PBPs) by forming a stable, covalent, acyl-enzyme complex. During the evolution from PBPs to Class A beta-lactamases, the beta-lactamases acquired Glu166 to activate a catalytic water and cleave the acyl-enzyme bond. Here we present three product complex crystal structures of CTX-M-14 Class A beta-lactamase with a ruthenocene-conjugated penicillin-a 0.85 angstrom resolution structure of E166A mutant complexed with the penilloate product, a 1.30 angstrom resolution complex structure of the same mutant with the penicilloate product, and a 1.18 angstrom resolution complex structure of S70G mutant with a penicilloate product epimer-shedding light on the catalytic mechanisms and product inhibition of PBPs and Class A beta-lactamases. The E166A-penilloate complex captured the hydrogen bonding network following the protonation of the leaving group and, for the first time, unambiguously show that the ring nitrogen donates a proton to Ser130, which in turn donates a proton to Lys73. These observations indicate that in the absence of Glu166, the equivalent lysine would be neutral in PBPs and therefore capable of serving as the general base to activate the catalytic serine. Together with previous results, this structure suggests a common proton relay network shared by Class A beta-lactamases and PBPs, from the catalytic serine to the lysine, and ultimately to the ring nitrogen. Additionally, the E166A-penicilloate complex reveals previously unseen conformational changes of key catalytic residues during the release of the product, and is the first structure to capture the hydrolyzed product in the presence of an unmutated catalytic serine.

  9. [Antimicrobial activity of fosfomycin under various conditions against standard strains, beta-lactam resistant strains, and multidrug efflux system mutants].

    Science.gov (United States)

    Mikuniya, Takeshi; Hiraishi, Toru; Maebashi, Kazunori; Ida, Takashi; Takata, Toshihiko; Hikida, Muneo; Yamada, Sakuo; Gotoh, Naomasa; Nishino, Takeshi

    2005-04-01

    The purpose of this study was to evaluate the possible benefit of fosfomycin (FOM) as prophylactic antibiotic in terms of antimicrobial activity and the potential of inducibility of beta-lactamase, compared with cefazolin, cefotiam, cefmetazole, and piperacillin that are commonly used as perioperative agents. The in vitro activity of FOM against aerobic Gram-negative bacteria using Mueller-Hinton agar or nutrient agar supplemented with glucose-6-phosphate (G6P) as tested medium increased within a range from 2 to 256 times the activity in the medium without G6P. However, the susceptibility of Gram-positive bacteria to FOM remained largely unchanged with or without G6P. There was no aerobic- or anaerobic-bacteria which changed susceptibility against beta-lactam antibiotics under various tested medium conditions. FOM demonstrated strong bactericidal activity against Escherichia coli and Pseudomonas aeruginosa in a dose dependent manner, and decreased viable cell counts of Staphylococcus aureus. In the case of P. aeruginosa, transmission electron micrographs study revealed that numerous lysed cells were present 2 hours after treatment with FOM at four times the MIC. First and second generation cephalosporins induced AmpC-type beta-lactamase in a dose dependent manner among beta-lactamase inducible strains of P. aeruginosa and Enterobacter cloacae. On the other hand, inducible activity of FOM on beta-lactamase production was less than 1/25 to 1/65 compared with those of cephalosporins. In addition, FOM maintained strong antimicrobial activity for over then 20 years after marketing, because of the excellent stability against various types of beta-lactamase produced by plasmid-carrying bacteria and clinical isolates. FOM was not extruded by four types of efflux systems, such as MexAB-OprM, MexCD-OprJ, MexXY/ OprM and MexEF-OprN, however beta-lactam antibiotics were substrates of MexAB-OprM and MexCD-OprJ. In conclusion, FOM provides adequate coverage for both aerobic Gram

  10. [Antibacterial activity of panipenem against clinical isolates in 2000 and 2001].

    Science.gov (United States)

    Abe, Tomomi; Fukuoka, Takashi; Sato, Yuki; Ito, Kazuyoshi; Sei, Masami

    2002-12-01

    As the post-marketing surveillance of panipenem/betamipron (Carbenin), MICs of panipenem (PAPM) against 1355 clinical isolates of 28 species from 15 medical institutions all over Japan from June 2000 to March 2001 were measured using the broth microdilution method approved by the Japanese Society of Chemotherapy and compared with those of parenteral carbapenem antibacterials, imipenem (IPM) and meropenem (MEPM), and parenteral cephem antibacterials, cefozopran, cefepime, and sulbactam/cefoperazone. The activity of PAPM was comparable to that of IPM against almost all species tested. Compared with MEPM, PAPM was more active against Gram-positive bacteria and Bacteroides spp., and less active against Gram-negative bacteria. Compared with the parenteral cephems, PAPM was more active against most of species tested and its MIC ranges were narrower than those of the cephems as were those of other carbapenems. In this surveillance study, the incidence of resistance in various species were as follows: 39.3% for methicillin-resistant Staphylococcus aureus, 47.3% for penicillin-intermediate Streptococcus pneumoniae (PISP), 15.1% for penicillin-resistant S. pneumoniae (PRSP), 0.9% for extended-spectrum beta-lactamase (ESBL) producing Escherichia coli, 3.4% for ESBL producing Klebsiella pneumoniae, 19.2% for beta-lactamase producing Haemophilus influenzae, 24.0% for beta-lactamase-negative ampicillin-resistant (BLNAR) H. influenzae, and 1.0% for metallo-beta-lactamase producing Pseudomonas aeruginosa. Against these resistant strains, carbapenems including PAPM showed generally more potent activity than cephems. It was noted that PAPM showed the most potent activity against PISP and PRSP, which showed high incidence of 62.4% totally, among tested drugs. Metallo-beta-lactamase producing P. aeruginosa exhibited high resistance and BLNAR H. influenzae also exhibited low susceptibility against all tested drugs. But no remarkable change in the activity of PAPM against other species

  11. Evaluation of Eight Different Cephalosporins for Detection of Cephalosporin Resistance in Salmonella enterica and Escherichia coli

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Hasman, Henrik; Veldman, K

    2010-01-01

    This study evaluates the efficacy of eight different cephalosporins for detection of cephalosporin resistance mediated by extended spectrum beta-lactamases (ESBL) and plasmidic AmpC beta-lactamases in Salmonella and Escherichia coli. A total of 138 E. coli and 86 Salmonella isolates with known beta......-resistant but cephalosporin-susceptible, 56 ESBL isolates and 19 isolates with plasmidic AmpC, as well as 10 ampC hyper-producing E. coli. The minimum inhibitory concentration distributions and zone inhibitions varied with the tested compound. Ampicillin-resistant isolates showed reduced susceptibility to the cephalosporins...... compared to ampicillin-susceptible isolates. Cefoperazone, cefquinome, and cefuroxime were not useful in detecting isolates with ESBL or plasmidic AmpC. The best substances for detection were cefotaxime, cefpodoxime, and ceftriaxone, whereas ceftazidime and ceftiofur were not as efficient. Ceftriaxone may...

  12. Analysis of OXA-204 carbapenemase-producing Enterobacteriaceae reveals possible endoscopy-associated transmission, France, 2012 to 2014.

    Science.gov (United States)

    Potron, Anaïs; Bernabeu, Sandrine; Cuzon, Gaëlle; Pontiès, Valérie; Blanchard, Hervé; Seringe, Elise; Naas, Thierry; Nordmann, Patrice; Dortet, Laurent

    2017-12-01

    OXA-48-like beta-lactamase producing bacteria are now endemic in several European and Mediterranean countries. Among this carbapenemase family, the OXA-48 and OXA-181 variants predominate, whereas other variants such as OXA-204 are rarely reported. Here, we report the molecular epidemiology of a collection of OXA-204-positive enterobacterial isolates (n = 29) recovered in France between October 2012 and May 2014. This study describes the first outbreak of OXA-204-producing Enterobacteriaceae in Europe, involving 12 isolates of an ST90 Escherichia coli clone and nine isolates of an ST147 Klebsiella pneumoniae clone. All isolates co-produced the cephalosporinase CMY-4, and 60% of them co-produced the extended-spectrum beta-lactamase CTX-M-15. The bla OXA-204 gene was located on a 150-kb IncA/C plasmid, isolated from various enterobacterial species in the same patient, indicating a high conjugative ability of this genetic vehicle.

  13. Differences in antimicrobial susceptibility of pigmented and unpigmented colonial variants of Mycobacterium avium.

    Science.gov (United States)

    Stormer, R S; Falkinham, J O

    1989-01-01

    Unpigmented colonial variants were isolated from pigmented Mycobacterium avium isolates recovered from patients with acquired immunodeficiency syndrome and the environment. The variants were interconvertible: the rate of transition from unpigmented to pigmented type was 4.0 x 10(-5) variants per cell per generation. The unpigmented variants were more tolerant to antibiotics, especially beta-lactams, and Cd2+ and Cu2+ salts than were their pigmented parents. Both pigmented and unpigmented variants of the strains produced beta-lactamase, although beta-lactamase did not appear to be a determinant of beta-lactam susceptibility. Pigmented variants grew more rapidly in a number of commonly used mycobacterial media, were more hydrophobic, and had higher carotenoid contents than their unpigmented segregants. PMID:2808669

  14. Klebsiella pneumoniae nueva Delhi metalo-betalactamasa en el hospital nacional Dos de Mayo. Lima, Perú

    OpenAIRE

    Resurrección-Delgado, Cristhian; Servicio de Enfermedades Infecciosas y Tropicales, Hospital Nacional Dos de Mayo. Lima, Perú. Médico infectólogo; Montenegro-Idrogo, Juan José; Servicio de Enfermedades Infecciosas y Tropicales, Hospital Nacional Dos de Mayo. Lima, Perú. Facultad de Medicina. Universidad Nacional Mayor de San Marcos. Lima, Perú. médico residente de Infectología; Chiappe-Gonzalez, Alfredo; Servicio de Enfermedades Infecciosas y Tropicales, Hospital Nacional Dos de Mayo. Lima, Perú. Médico infectólogo; Vargas-Gonzales, Renzo; Servicio de Enfermedades Infecciosas y Tropicales, Hospital Nacional Dos de Mayo. Lima, Perú. Facultad de Medicina. Universidad Nacional Mayor de San Marcos. Lima, Perú. médico residente de Infectología; Cucho-Espinoza, Carolina; Servicio de Microbiología, Hospital Nacional Dos de Mayo. Lima, Perú. médico patólogo clínico; Mamani-Condori, Dick Henry; Servicio de Microbiología, Hospital Nacional Dos de Mayo. Lima, Perú. tecnólogo médico; Huaroto-Valdivia, Luz María; Servicio de Microbiología, Hospital Nacional Dos de Mayo. Lima, Perú. médico patólogo clínico

    2017-01-01

    The emergence of Enterobacteria producing carbapenemases of type New Delhi Metalo beta-lactamases (NDM), represent, today, a real problem of world public health. The presence of this resistance mechanism limits or nullifies the therapeutic options to combat these bacteria. In Latin America, the figures are getting higher, as they are reported in Guatemala, Colombia, Chile, Argentina, among others. Peru has not, to date, described the presence of this resistance pattern; however for several ye...

  15. Decreased outer membrane permeability in imipenem-resistant mutants of Pseudomonas aeruginosa.

    OpenAIRE

    Trias, J; Dufresne, J; Levesque, R C; Nikaido, H

    1989-01-01

    The outer membrane of imipenem-resistant mutants of Pseudomonas aeruginosa was shown to have decreased permeability to imipenem but not to cephaloridine. These experiments were performed with intact cells and liposomes containing imipenem-hydrolyzing beta-lactamase derived from Pseudomonas maltophilia, in both cases utilizing an imipenem concentration of 50 microM. In contrast, liposome swelling assays using imipenem at 8 mM detected no significant difference between the imipenem-resistant mu...

  16. Priorities in the prevention and control of multidrug-resistant Enterobacteriaceae in hospitals.

    LENUS (Irish Health Repository)

    Khan, A S

    2012-10-01

    Multidrug-resistant Enterobacteriaceae (MDE) are a major public health threat due to international spread and few options for treatment. Furthermore, unlike meticillin-resistant Staphylococcus aureus (MRSA), MDE encompass several genera and multiple resistance mechanisms, including extended-spectrum beta-lactamases and carbapenemases, which complicate detection in the routine diagnostic laboratory. Current measures to contain spread in many hospitals are somewhat ad hoc as there are no formal national or international guidelines.

  17. Examination pf Potential Anti-Tumor Activity of N-Thiolated B-Lactam Antibiotics in Nude Mice Bearing Human Breast Tumors

    Science.gov (United States)

    2005-08-01

    their mechanism of action, production of reactive oxygen species (ROS), which leads to toxicity of the cardiomyocytes and subsequent chronic and acute...gemcitabine, and the purine analogs, such as 6- mercaptopurine and 6-thioguanine. These analogs substitute for nucleic acid bases in both DNA and RNA...Bioorg Med Chem Lett 12, 2229-2231 (2002) 14. Holt, R.J. and G.T. Stewart: Production of Amidase and beta-Lactamase by Bacteria. J Gen Microbiol 36

  18. Next-Generation Sequencing for Typing and Detection of ESBL and MBL E. coli causing UTI

    OpenAIRE

    Nabakishore Nayak; Mahesh Chanda Sahu

    2017-01-01

    Next-generation sequencing (NGS) has the potential to provide typing results and detect resistance genes in a single assay, thus guiding timely treatment decisions and allowing rapid tracking of transmission of resistant clones. We can be evaluated the performance of a new NGS assay during an outbreak of sequence type 131 (ST131) Escherichia coli infections in a teaching hospital. The assay will be performed on 100 extended-spectrum- beta-lactamase (ESBL) E. coli isolates collected from UTI d...

  19. MICROBIOLOGICAL CHARACTERISATION OF Haemophilus influenzae STRAINS ISOLATED FROM PATIENTS WITH INVASIVE AND RESPIRATORY DISEASES

    Directory of Open Access Journals (Sweden)

    Tomislav Kostyanev

    2010-01-01

    Full Text Available A total of 175 H. influenzae strains were collected between 1994 and 2009 from all aged patient groups. The strains were isolated from patients with invasive and community-acquired respiratory tract infections. All strains were identified according to standard microbiological methods. Serotyping was done by a coagglutination test and by molecular PCR capsular genotyping. Beta-lactamase production was determined by the chromogenic cephalosporin test with nitrocephin as substrate. Most of the isolated H. influenzae strains were from children under 5 years of age (57.7%. Overall, 61 strains belonged to serotype b (34.9% by the means of PCR capsular typing, 1 strain was type f, and 113 isolates (64.6% were non-typeable (non-encapsulated H. influenzae. Among the infants and children with meningitis or other invasive infections, aged 2 month to 5 years, all strains, except one, were serotype b. In respiratory tract infections (pneumonia, otitis media, sinusitis and people with chronic pulmonary diseases - exacerbations of COPD, bronchiectasis, cystic fibrosis the most common - 96.5% were non-typeable strains in both groups children and adults. Overall, the prevalence of beta-lactamase production was 19.4%. But, it was much higher for invasive strains from CSF isolates - 37.7%, 25% in blood samples, and 37.5% in otitis media causative strains. Beta-lactamase production was less frequent in respiratory tract isolates - in sputum 13.3% and in URT samples - 2.3%. The rate of beta-lactamase production in CSF isolates has not changed for the last 10 years.PCR capsular genotyping method has to be performed for all non-b-type strains. The implementation of Hib vaccine in our country will be accompanied by a reduction in invasive diseases caused by H. influenzae type b in children, but it is not useful in preventing infections caused by non-typeable H. influenzae strains.

  20. Recruitment of a penicillin-binding protein gene from Neisseria flavescens during the emergence of penicillin resistance in Neisseria meningitidis

    OpenAIRE

    SPRATT, BG; ZHANG, QY; JONES, DM; HUTCHISON, A; BRANNIGAN, JA; DOWSON, CG

    1989-01-01

    Non-beta-lactamase-producing, penicillin-resistant strains of Neisseria meningitidis produce altered forms of penicillin-binding protein 2 that have decreased affinity for penicillin. The sequence of the penicillin-binding protein 2 gene (penA) from a penicillin-resistant strain of N. meningitidis was compared to the sequence of the same gene from penicillin-sensitive strains and from penicillin-sensitive and penicillin-resistant strains of Neisseria gonorrhoeae. The penA genes from penicilli...

  1. Mass Spectrometry to Identify New Biomarkers of Nerve Agent Exposure

    Science.gov (United States)

    2009-04-01

    calculations for class C beta- lactamases: the role of Tyr-150. Proteins 40:23-28. Lewis SD, Johnson FA and Shafer JA (1976) Potentiometric determination of...example, in the presence of a 2-fold excess of soman, 100% inactivation of the AAA activity of albumin was never reached. The titration plot (not shown... Titration of trans- ferrin with ferric ion was performed as described (Welch and Skinner, 1989). A 1 mg/ml solution of apo-human transferrin was

  2. Technical note: Antimicrobial susceptibility of Portuguese isolates of Staphylococcus aureus and Staphylococcus epidermidis in subclinical bovine mastitis.

    Science.gov (United States)

    Nunes, S F; Bexiga, R; Cavaco, L M; Vilela, C L

    2007-07-01

    To evaluate the antimicrobial resistance traits of staphylococci responsible for subclinical bovine mastitis in Portugal, the minimum inhibitory concentrations (MIC) of 7 antimicrobial agents, frequently administered for mastitis treatment, were determined for 30 Staphylococcus aureus and 31 Staphylococcus epidermidis field isolates. Beta-lactamase production was detected through the use of nitrocefin-impregnated discs. The MIC that inhibited 90% of the isolates tested (MIC90) of penicillin, oxacillin, cefazolin, gentamicin, sulfamethoxazole/trimethoprim, oxytetracycline, and enrofloxacin were, respectively, 4, 0.5, 1, 1, 0.25, 0.25, and 0.06 microg/mL for Staph. aureus and > or = 64, 8, 1, 32, > or = 64, > or = 64, and 0.06 microg/mL for Staph. epidermidis. All Staph. aureus isolates showed susceptibility to oxacillin, cefazolin, gentamicin, sulphamethoxazole/trimethoprim, and enrofloxacin. Beta-lactamase production was detected in 20 of these isolates (66.7%), all of which were resistant to penicillin. Of the 31 Staph. epidermidis tested, 24 (77.4%) were beta-lactamase positive. All isolates were susceptible to both cefazolin and enrofloxacin. Nine Staph. epidermidis isolates were resistant to oxacillin, with MIC values ranging from 4 to 8 microg/mL. The MIC values of 5 antimicrobial agents tested were higher than those reported in other countries. Enrofloxacin was the only exception, showing lower MIC values compared with other reports. Overall, the antimicrobial agents tested in our study, with the exception of penicillin, were active against the 61 isolates studied.

  3. [Epidemiologic diagnostic of nosocomial suppurative-septic infections of Pseudomonas etiology based on intraspecies typing of causative agent].

    Science.gov (United States)

    Fel'dblium, I V; Zakharova, Iu A; Nikolaeva, A M; Fedotova, O S

    2013-01-01

    Scientific justification of optimization of epidemiologic diagnostic of suppurative-septic infection (SSI) caused by Pseudomonas aeruginosa based on comparability of antibiotic sensitivity and beta-lactamase production. Intraspecies typing of 37 P. aeruginosa strains isolated during microbiological monitoring of 106 patients and 131 objects of clinical environment of surgical and obstetrician hospitals by using a complex ofphenotypic and molecular-biological methods including determination of sensitivity to antibiotics by serial dilutions method and PCR-diagnostics with determination of TEM, SHV, CTX, OXA, MBL, VIM genes was performed. P. aeruginosa strains combined into groups by isolation location during studies turned out to be heterogeneous by sensitivity to antibiotics and beta-lactamase production that allowed to form subgroups of strains by focality attribute. Isolates recovered from different SSI foci had significant differences in minimal inhibitory concentration (MIC) reaching 1024 times. MIC parameter within subgroups did not exceed 8 - 16 consequent dilutions. Use of a complex of phenotypic and molecular-biologic methods of causative agent typing including determination of sensitivity to antibiotics by serial dilutions method and evaluation of beta-lactamase production allowed to establish a mechanism of development of SSI epidemic process caused by P. aeruginosa, detect origins and reservoirs of infection in hospital, modes and factors of transmission and reach maximum justification of epidemiologic control and prophylaxis measures of localization of foci of nosocomial infections of pseudomonas etiology.

  4. Dissemination of a Multidrug-Resistant VIM-1- and CMY-99-Producing Proteus mirabilis Clone in Bulgaria.

    Science.gov (United States)

    Markovska, Rumyana; Schneider, Ines; Keuleyan, Emma; Ivanova, Dobrinka; Lesseva, Magdalena; Stoeva, Temenuga; Sredkova, Mariya; Bauernfeind, Adolf; Mitov, Ivan

    2017-04-01

    The aim of this study was to analyze the beta-lactamases and the molecular epidemiology of 19 clinically significant isolates of Proteus mirabilis with decreased susceptibility to imipenem, which have been collected from seven hospitals, located in different Bulgarian towns (Sofia, Varna, and Pleven). The isolates were obtained from blood, urine, tracheal and wound specimens. One additional isolate from hospital environment was included. Susceptibility testing, conjugation experiments, and plasmid replicon typing were carried out. Beta-lactamases were characterized by isoelectric focusing, PCR, and sequencing. Clonal relatedness was investigated by RAPD and PFGE. Integron mapping was performed by PCR and sequencing. All isolates showed a multidrug-resistance profile, but remained susceptible to piperacillin/tazobactam, cefepime, meropenem, and fosfomycin. They produced identical beta-lactamases, namely: TEM-1, VIM-1, and CMY-99. PCR mapping revealed that the bla VIM-1 gene was part of a class 1 integron that additionally included the aac(6')-I, dhfrA1, and ant(3″)-Ia genes. In addition, 17 of the isolates carried the armA gene. Conjugation experiments and plasmid replicon typing were unsuccessful. The isolates were clonally related according to RAPD and PFGE typing. This study reveals the nationwide distribution of a multidrug-resistant P. mirabilis clone producing VIM-1 and CMY-99 along with the presence of different aminoglycoside resistance mechanisms.

  5. Development of a multiplex PCR assay for the detection and differentiation of Burkholderia pseudomallei, Burkholderia mallei, Burkholderia thailandensis, and Burkholderia cepacia complex.

    Science.gov (United States)

    Zakharova, Irina; Teteryatnikova, Natalya; Toporkov, Andrey; Viktorov, Dmitry

    2017-10-01

    Two species of Burkholderia pseudomallei complex (Bpc), B. pseudomallei and B. mallei, can cause severe life-threatening infections. Rapidly discerning individual species within the group and separating them from other opportunistic pathogens of the Burkholderia cepacia complex (Bcc) is essential to establish a correct diagnosis and for epidemiological surveillance. In this study, a multiplex PCR assay based on the detection of an individual set of chromosomal beta-lactamase genes for single-step identification and differentiation of B. pseudomallei, B. mallei, B. thailandensis, and Bcc was developed. Two pairs of primers specific to a distinct class of B metallo-beta-lactamase genes and a pair of primers specific to the oxacillin-hydrolyzing class D beta-lactamase gene were demonstrated to successfully discriminate species within Bpc and from Bcc. The assay sensitivity was 9561 genomic equivalents (GE) for B. pseudomallei, 7827 GE for B. mallei, 8749 GE for B. thailandensis and 6023 GE for B. cepacia. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Prevalence of beta-lactams resistance among Escherichia coli clinical isolates from a hospital in Algiers.

    Science.gov (United States)

    Messai, Y; Benhassine, T; Naim, M; Paul, G; Bakour, R

    2006-06-01

    A high prevalence of beta-lactams resistance among Enterobacteriaceae have been reported worldwide; however, there are not sufficient data on this issue in Algeria. beta-Lactams susceptibility of 203 Escherichia coli clinical isolates was determined by agar diffusion method, and production of extended-spectrum beta-lactamases (ESBL) was screened by double-disk synergy test. This analysis showed five well-defined phenotypes: 1) 62 isolates (30.5%) were susceptible to all beta-lactams; 2) 135 isolates (66.5%) presented a broad-spectrum beta-lactamases phenotype (BSBL); 3) three isolates (1.5%) were defined as producing ESBLs; 4) two isolates (1%) were AmpC cephalosporinase producers; and 5) one isolate (0.5%) presented a phenotype of cell-decreased permeability to beta-lactams. Isoelectric focusing revealed beta-lactamases with isolectric points of 5.4 or 7.6 for isolates with BSBL phenotype; approximately 9.0 for two ESBL isolates; 5.4, 7.6 and approximately 9.0 for the remaining ESBL isolate; and 5.4 and approximately 9.0 for the AmpC isolates. The cefotaxime hydrolysis corresponds to the basic bands with an isoelectric point of approximately 9.0. Conjugation assay showed transfer of penicillinase and AmpC resistance phenotypes and their corresponding beta-lactamases to recipient E. coli BM21 in association with plasmids of 71.4 kb for the AmpC isolates and from 40-56 kb for penicillinase isolates. This result showed that the AmpC phenotype is plasmid mediated. ESBL isolates were found not to transfer their resistance through conjugation experiment. Polymerase chain reaction (PCR) experiments using primers specific to blaTEM, blaAmpC and blaCTX-M genes showed specific amplification with blaCTX-M primer for two ESBL isolates; blaTEM and blaCTX-M for the remaining ESBL isolate; and blaTEM and blaAmpC for the AmpC isolates and their corresponding transconjugants. The study showed a high rate of isolates producing penicillinase, and low frequencies of AmpC and ESBL

  7. Detection of ctx-M gene in ESBL-producing E. coli strains isolated from urinary tract infection in Semnan, Iran.

    Science.gov (United States)

    Tabar, Mahbobeh Mohammad; Mirkalantari, Shiva; Amoli, Rabeeh Izadi

    2016-07-01

    The incidence of urinary tract infections caused by Extended-Spectrum Beta Lactamase (ESBL) producing Escherichia coli (E. coli) strains due to long term and overuse of broad-spectrum cephalosporine is on the rise. CTX beta-lactamase type, a broad-spectrum beta-lactamase, has been expanding in many countries. The ctx gene is harbored on a plasmid that is spread between Enterobacteriaceae family, especially in E. coli. The aim of this study was to determine the pattern of antimicrobial resistance and investigate the prevalent ESBL phenotype and the ctx-M gene in E. coli isolated from patients with urinary tract infections (UTI) in Semnan. A cross sectional study was performed on 109 strains of E. coli isolated from the urine culture of patient suffering from a UTI referred to Shafa hospital (Semnan, Iran) during March-July 2015. Antimicrobial susceptibility testing was applied and the prevalence of the ESBL phenotype was confirmed using combination disk. PCR methods were completed for amplification of the bla ctx gene. Data were analyzed using SPSS version 18 software. One hundred ninety samples (4.16%) were identified as E. coli. Twenty one (26.6%) of E. coli were ESBL positive and 73.4% were ESBL negative. There was 100% susceptibility to imipeneme. Twenty (68.97%) out of 29 isolates were positive for the ctx-M gene, as detected by PCR. In urinary tract infections, antibiotic treatment was experimental and detailed information regarding the sensitivity of bacteria in the area can be useful to achieve the best treatment.

  8. Yersinia enterocolitica targets cells of the innate and adaptive immune system by injection of Yops in a mouse infection model.

    Directory of Open Access Journals (Sweden)

    Martin Köberle

    2009-08-01

    Full Text Available Yersinia enterocolitica (Ye evades the immune system of the host by injection of Yersinia outer proteins (Yops via a type three secretion system into host cells. In this study, a reporter system comprising a YopE-beta-lactamase hybrid protein and a fluorescent staining sensitive to beta-lactamase cleavage was used to track Yop injection in cell culture and in an experimental Ye mouse infection model. Experiments with GD25, GD25-beta1A, and HeLa cells demonstrated that beta1-integrins and RhoGTPases play a role for Yop injection. As demonstrated by infection of splenocyte suspensions in vitro, injection of Yops appears to occur randomly into all types of leukocytes. In contrast, upon infection of mice, Yop injection was detected in 13% of F4/80(+, 11% of CD11c(+, 7% of CD49b(+, 5% of Gr1(+ cells, 2.3% of CD19(+, and 2.6% of CD3(+ cells. Taking the different abundance of these cell types in the spleen into account, the highest total number of Yop-injected cells represents B cells, particularly CD19(+CD21(+CD23(+ follicular B cells, followed by neutrophils, dendritic cells, and macrophages, suggesting a distinct cellular tropism of Ye. Yop-injected B cells displayed a significantly increased expression of CD69 compared to non-Yop-injected B cells, indicating activation of these cells by Ye. Infection of IFN-gammaR (receptor- and TNFRp55-deficient mice resulted in increased numbers of Yop-injected spleen cells for yet unknown reasons. The YopE-beta-lactamase hybrid protein reporter system provides new insights into the modulation of host cell and immune responses by Ye Yops.

  9. Structures of the Class D Carbapenemase OXA-24 from Acinetobacter baumannii in Complex with Doripenem

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, Kyle D.; Ortega, Caleb J.; Renck, Nicholas A.; Bonomo, Robert A.; Powers, Rachel A.; Leonard, David A. (Case Western); (Grand Valley)

    2012-02-08

    The emergence of class D {beta}-lactamases with carbapenemase activity presents an enormous challenge to health practitioners, particularly with regard to the treatment of infections caused by Gram-negative pathogens such as Acinetobacter baumannii. Unfortunately, class D {beta}-lactamases with carbapenemase activity are resistant to {beta}-lactamase inhibitors. To better understand the details of the how these enzymes bind and hydrolyze carbapenems, we have determined the structures of two deacylation-deficient variants (K84D and V130D) of the class D carbapenemase OXA-24 with doripenem bound as a covalent acyl-enzyme intermediate. Doripenem adopts essentially the same configuration in both OXA-24 variant structures, but varies significantly when compared to the non-carbapenemase class D member OXA-1/doripenem complex. The alcohol of the 6a hydroxyethyl moiety is directed away from the general base carboxy-K84, with implications for activation of the deacylating water. The tunnel formed by the Y112/M223 bridge in the apo form of OXA-24 is largely unchanged by the binding of doripenem. The presence of this bridge, however, causes the distal pyrrolidine/sulfonamide group to bind in a drastically different conformation compared to doripenem bound to OXA-1. The resulting difference in the position of the side-chain bridge sulfur of doripenem is consistent with the hypothesis that the tautomeric state of the pyrroline ring contributes to the different carbapenem hydrolysis rates of OXA-1 and OXA-24. These findings represent a snapshot of a key step in the catalytic mechanism of an important class D enzyme, and might be useful for the design of novel inhibitors.

  10. Investigation of the antibiotic susceptibility patterns of pathogens causing nosocomial infections

    International Nuclear Information System (INIS)

    Yaman, Akgun; Kibar, Filiz; Buyukcelik, Ozlem; Tasova, Yesim; Inal, A.S.; Saltoglu, Nese; Kurtaran, Behice; Dundal, Ismail H.

    2004-01-01

    The aim of this study is to determine the resistance patterns of bacteria causing nosocomial infections. The outcome of this resistance was followed for 3 years. This study was carried out during 2000 to 2002 at a university hospital in Turkey. The resistance patterns of 570 bacteria (390 Gram-negative, 180 Gram-positive) against meropenem, imipenem, ceftazidime, cefotaxime, cefepime, piperacillin/tazobactam, ciprofloxacin and tobramycin were investigated using the E-test. Extended-spectrum beta-lactamase (ESBL) production was determined using ceftazidime and ceftazidime/clavulanic acid E-test strips. Meropenem was the most effective antibiotic against Gram-negative organisms (89.0%); this was followed by imipenem (87.2%) and piperacillin/tazobactam (66.4%). The most active antibiotic against Gram-positive bacteria was imipenem (87.2%) and this was followed by piperacillin/tazobactam (81.7%) and meropenem (77.8%). The rates of production of ESBL by Escherichia coli were 20.9%, Klebsiella pneumoniae 50% and Serratia marcescens were 46.7%. Extended-spectrum beta-lactamase production increased each year (21.7%, 22.1% and 45.5%). All of the ESBL producing isolates were sensitive to meropenem and 98.5% sensitive to imipenem. AmpC beta-lactamase was produced by 20.9% of the Enterobacter species spp, Citrobacter spp. and Serratia marcescens. All of these were sensitive to meropenem and 77.8% to imipenem and ciprofloxacin. Multi-drug resistance rates in Acinetobacter spp were 45.4% and 37.7% in Pseudomonas aeruginosa isolates. As in the entire world, resistance to antibiotics is a serious problem in our country. Solving of this problem depends primarily on prevention of the development of resistance. (author)

  11. Characterization of ESBL-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urine of nonhospitalized patients in the Zagreb region

    Directory of Open Access Journals (Sweden)

    Branka Bedenić,

    2010-02-01

    Full Text Available Aim To determine the prevalence of ESBL-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urine of nonhospitalized patients during a three-year period, to determine their antibiotic susceptibility, investigate the transfer of ESBL genes with cotransfer of resistance and to characterize isolated beta-lactamases. Methods Antimicrobial susceptibility was determined by disk diffusion and broth microdilution methods. The double-disk test was used for ESBL detection. Transfer of resistance was performed by broth mating method and characterization of isolated beta-lactamases by polymerase chain reaction. Results The prevalence of ESBL-producing E. coli was 1.5% and of K. pneumoniae 4.1% with its different distribution according to patients`age and gender. ESBL-producing K. pneumoniae showed high resistance rates to aminoglycosides, cotrimoxazole, nitrofurantoin and quinolones while ESBL-producing E. coli isolates, with exception of high aminoglycoside resistance, showed low resistance rates to other antibiotics. Successful conjugation of ESBL genes was obtained with 25% E. coli and 76.2% K. pneumoniae strains. Comparing to E. coli, K. pneumoniae strains showed higher rates of aminoglycosideand cotrimoxazole resistance cotransfer. Beta-lactamases of investigated strains belonged to TEM, SHV and CTX-M families.Conclusion The existence of multiple-resistant ESBL-producing E. coli and K. pneumoniae strains was confirmed in observed outpatient population. ESBL-producing K. pneumoniae isolates, in contrast toESBL-producing E. coli, showed higher resistance rates to non-beta-lactam antibiotics, probably caused by cotransfer of resistance genes located on the same plasmid as ESBL genes. It is important to monitor the prevalence of such strains and their possible spreading in the outpatient population of the Zagreb region

  12. Temporal Trends in Antimicrobial Resistance and Virulence-Associated Traits within the Escherichia coli Sequence Type 131 Clonal Group and Its H30 and H30-Rx Subclones, 1968 to 2012

    DEFF Research Database (Denmark)

    Olesen, Bente; Frimodt-Møller, Jakob; Leihof, Rikke Fleron

    2014-01-01

    occurred only after 2000 and 2002, respectively, in association with the H30 and H30-Rx subclones, which were characterized by multidrug resistance (including extended-spectrum-beta-lactamase [ESBL] production: H30-Rx) and absence of biofilm production. Capsular antigen K100 occurred exclusively among H30......-Rx ST131 subclones, rather than representing acquisition of resistance by diverse ST131 strains. Distinctive characteristics of the H30-Rx subclone-including specific virulence genes (iutA, afa and dra, kpsII), the K100 capsule, multidrug resistance, and ESBL production-possibly contributed...

  13. Prevalence and molecular characterization of clinical isolates of Escherichia coli expressing an AmpC phenotype

    DEFF Research Database (Denmark)

    Jørgensen, Rikke Lind; Nielsen, Jesper Boye; Friis-Møller, Alice

    2010-01-01

    . Hyperproduction of AmpC beta-lactamase was confirmed by isoelectric focusing (IEF). The presence of a plasmid-mediated ampC gene (pAmpC) was detected by multiplex PCR. The promoter and the entire reading frame of the chromosomal ampC gene were sequenced to identify promoter mutations associated...... by multilocus sequence typing (MLST). The remaining isolates all had mutations or insertions in the promoter region, which could explain increased expression of the chromosomal AmpC enzyme. Mutations in the ampC gene associated with extended activity were rare and did not cause resistance to cefepime...

  14. Antimicrobial susceptibility profile of Neisseria gonorrhoeae at STI clinic

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    Shilpee C

    2008-01-01

    Full Text Available A total of 100 consecutive patients who attended a sexually transmitted infections clinic were studied. Thirteen had gonococcal urethritis, of which 10 showed growth of Neisseria gonorrhoeae on culture. All the isolates were tested for antimicrobial susceptibility by Australian Gonococcal Surveillance Programme (AGSP method and beta lactamase production by chromogenic cephalosporin test. Four patients were co-infected with each of the following: HIV, HBV and Chlamydia trachomatis . Gonococcal urethritis (13% was found more in male patients. Ten percent gonococcal isolates were penicillinase-producing N. gonorrhoeae , and another 10% were tetracycline-resistant N. gonorrhoeae .

  15. Antimicrobial susceptibility of coagulase-negative Staphylococcus species isolated from bovine milk.

    Science.gov (United States)

    Sawant, A A; Gillespie, B E; Oliver, S P

    2009-02-16

    Coagulase-negative Staphylococcus (CNS) isolates (n=168) obtained from milk from heifers and dairy cows were screened for minimum inhibitory concentration (MIC) to antimicrobials used commonly for mastitis therapy. Of the 10 CNS species included in the study, the predominant species were Staphylococcus chromogenes (n=61), Staphylococcus epidermidis (n=37), Staphylococcus hyicus (n=37), and Staphylococcus simulans (n=16). The majority of CNS was susceptible to ampicillin, oxacillin, cephalothin, and ceftiofur. Erythromycin and pirlimycin were also very effective in vitro inhibitors of CNS. The only exception was observed with S. epidermidis. Of 37 S. epidermidis evaluated, 13 (35%) exhibited efflux-based resistance to erythromycin (> or =16 microg/ml) encoded by msrA and one isolate carried ermC encoding ribosomal methylase-based resistance to both erythromycin (> or =64 microg/ml) and pirlimycin (> or =64 microg/ml). A total of 17 S. epidermidis, 11 S. chromogenes, and one S. hyicus exhibited phenotypic resistance to ampicillin (> or =0.5 microg/ml). Constitutive beta-lactamase production was observed in all ampicillin resistant isolates except 4 S. epidermidis that exhibited inducible beta-lactamase production. Induced beta-lactamase production was also observed in 13 S. epidermidis that were phenotypically susceptible to the entire MIC panel. All isolates that produced beta-lactamase either constitutively or by induction carried blaZ. S. epidermidis (n=12, 32%) that were resistant to methicillin (oxacillin > or =0.5 microg/ml) carried low affinity penicillin-binding protein encoded by mecA. Most multi-drug resistant (MDR) S. epidermidis (> or =2 resistance genes) were resistant to ampicillin, erythromycin and methicillin. All except one MDR S. epidermidis had icaAB, which encodes for polysaccharide intercellular adhesion. Based on pulsed field gel electrophoresis, MDR S. epidermidis were closely related genotypically, and were isolated from different cows on the

  16. THE NEW METALL-BETA-LACTAMASE’S INHIBITOR EFFICACY IN A MODEL SYSTEM IN VITRO

    Directory of Open Access Journals (Sweden)

    A. G. Afinogenova

    2016-01-01

    Full Text Available The Enterobacteriaceae antibiotics resistance depends on a combination of several mechanisms, such as the beta-lactamases overproduction, the microbial cell reduction outer membrane permeability (usually associated with loss of protein porin, the presence of efflux systems. Particularly noteworthy are the metallo-beta-lactamases (MBL whose presence causes resistance of gram-negative microorganisms to all beta-lactam antibiotics (in some cases except aztreonam. Currently there are no MBL inhibitors permitted for use in the clinic. The effective inhibitors search for carbapenem-resistant bacteria’ MBL authorized for use in the clinic and reinforcing effects of carbapenems, served as the basis for the present study. The work was carried out in three stages: 1 creating a model system using a standard enzyme reagent metallo-beta-lactamase P. aeruginosa recombinant expressed in E. coli, to evaluate the increasing of minimal inhibitory concentrations (MIC of carbapenems against previously sensitive Gram-negative microorganisms strains in vitro; 2 evaluation of MBL promising inhibitors in the presence of the same standard enzyme reagent; 3 evaluation of the ability of the identified inhibitors increase the carbapenems effects against clinical isolates of Gram-negative microorganisms producing MBL, in terms of the their MIC and fractional inhibitory concentration index (FIC index. The checkerboard array was modified to evaluate the combined use of carbapenems and potential MBL inhibitor — a drug from the group of bisphosphonates — etidronic acid. Using a standard enzyme reagent metallo-beta-lactamase P. aeruginosa recombinant expressed in E. coli, we created a model system that allows to assess the prospects of new inhibitors MBL gram-negative microorganisms. A dose-dependent effect of increasing the meropenem level MIC from reagent MBL quantity in a model system against previously antibiotic sensitive reference strains of microorganisms was

  17. E. coli bacteremia in comparison to K. pneumoniae bacteremia: influence of pathogen species and ESBL production on 7-day mortality

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    R. Leistner

    2016-10-01

    Full Text Available Abstract In a previous study, we demonstrated prolonged length of hospital stay in cases of extended-spectrum beta-lactamase (ESBL-positive K. pneumoniae bacteremia compared to bacteremia cases due to E. coli (ESBL-positive and –negative and ESBL-negative K. pneumoniae. The overall mortality was significantly higher in bacteremia cases resulting from ESBL-positive pathogens but also in K. pneumoniae cases disregarding ESBL-production. In order to examine whether pathogen species rather than multidrug resistance might affect mortality risk, we reanalyzed our dataset that includes 1.851 cases of bacteremia.

  18. Enterobacter Asburiae Pneumonia with Cavitation

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Seung Woo; Heo, Jeong Nam; Park, Choong Ki [Dept. of Radiology, Hanyang University College of Medicine, Guri Hospital, Guri (Korea, Republic of); Choi, Yo Won; Jeon, Seok Chol [Dept. of Radiology, Hanyang University College of Medicine, Seoul Hospital, Seoul (Korea, Republic of)

    2013-03-15

    Enterobacter species have increasingly been identified as pathogens over the past several decades. These bacterial species have become more important because most are resistant to cephalothin and cefoxitin, and can produce extended-spectrum {beta}-lactamase. Enterobacter asburiae (E. asburiae) is a gram-negative rod of the family Enterobacteriaceae, named in 1986. Since then, there has been only one clinical report of E. asburiae pneumonia. We report a case of E. asburiae pneumonia with cavitation and compare it with the previous case.

  19. Genetic markers associated with resistance to beta-lactam and quinolone antimicrobials in non-typhoidal Salmonella isolates from humans and animals in central Ethiopia

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    Tadesse Eguale

    2017-01-01

    Full Text Available Abstract Background Beta-lactam and quinolone antimicrobials are commonly used for treatment of infections caused by non-typhoidal Salmonella (NTS and other pathogens. Resistance to these classes of antimicrobials has increased significantly in the recent years. However, little is known on the genetic basis of resistance to these drugs in Salmonella isolates from Ethiopia. Methods Salmonella isolates with reduced susceptibility to beta-lactams (n = 43 were tested for genes encoding for beta-lactamase enzymes, and those resistant to quinolones (n = 29 for mutations in the quinolone resistance determining region (QRDR as well as plasmid mediated quinolone resistance (PMQR genes using PCR and sequencing. Results Beta-lactamase genes (bla were detected in 34 (79.1% of the isolates. The dominant bla gene was blaTEM, recovered from 33 (76.7% of the isolates, majority being TEM-1 (24, 72.7% followed by TEM-57, (10, 30.3%. The blaOXA-10 and blaCTX-M-15 were detected only in a single S. Concord human isolate. Double substitutions in gyrA (Ser83-Phe + Asp87-Gly as well as parC (Thr57-Ser + Ser80-Ile subunits of the quinolone resistance determining region (QRDR were detected in all S. Kentucky isolates with high level resistance to both nalidixic acid and ciprofloxacin. Single amino acid substitutions, Ser83-Phe (n = 4 and Ser83-Tyr (n = 1 were also detected in the gyrA gene. An isolate of S. Miami susceptible to nalidixic acid but intermediately resistant to ciprofloxacin had Thr57-Ser and an additional novel mutation (Tyr83-Phe in the parC gene. Plasmid mediated quinolone resistance (PMQR genes investigated were not detected in any of the isolates. In some isolates with decreased susceptibility to ciprofloxacin and/or nalidixic acid, no mutations in QRDR or PMQR genes were detected. Over half of the quinolone resistant isolates in the current study 17 (58.6% were also resistant to at least one of the beta-lactam antimicrobials

  20. Co-occurrence of colistin-resistance genes mcr-1 and mcr-3 among multidrug-resistant Escherichia coli isolated from cattle, Spain, September 2015.

    Science.gov (United States)

    Hernández, Marta; Iglesias, M Rocío; Rodríguez-Lázaro, David; Gallardo, Alejandro; Quijada, Narciso; Miguela-Villoldo, Pedro; Campos, Maria Jorge; Píriz, Segundo; López-Orozco, Gema; de Frutos, Cristina; Sáez, José Luis; Ugarte-Ruiz, María; Domínguez, Lucas; Quesada, Alberto

    2017-08-03

    Colistin resistance genes mcr-3 and mcr-1 have been detected in an Escherichia coli isolate from cattle faeces in a Spanish slaughterhouse in 2015. The sequences of both genes hybridised to same plasmid band of ca 250 kb, although colistin resistance was non-mobilisable. The isolate was producing extended-spectrum beta-lactamases and belonged to serotype O9:H10 and sequence type ST533. Here we report an mcr-3 gene detected in Europe following earlier reports from Asia and the United States. This article is copyright of The Authors, 2017.

  1. Treatment Modalities and Antimicrobial Stewardship Initiatives in the Management of Intra-Abdominal Infections

    Directory of Open Access Journals (Sweden)

    Charles Hoffmann

    2016-02-01

    Full Text Available Antimicrobial stewardship programs (ASPs focus on improving the utilization of broad spectrum antibiotics to decrease the incidence of multidrug-resistant Gram positive and Gram negative pathogens. Hospital admission for both medical and surgical intra-abdominal infections (IAIs commonly results in the empiric use of broad spectrum antibiotics such as fluoroquinolones, beta-lactam beta-lactamase inhibitors, and carbapenems that can select for resistant organisms. This review will discuss the management of uncomplicated and complicated IAIs as well as highlight stewardship initiatives focusing on the proper use of broad spectrum antibiotics.

  2. Determinação da produção de metalo-b-lactamases em amostras de Pseudomonas aeruginosa isoladas em João Pessoa, Paraíba

    OpenAIRE

    Santos Filho Lauro; Santos Isabele Beserra; Assis Alexandro Mangueira Lima de; Xavier Danilo Elias

    2002-01-01

    Bactérias produtoras de metalo-beta-lactamases (MBLs) são em grande parte resistentes aos betalactâmicos de largo espectro, incluindo oximino-aminotiazol cefalosporinas e também aos carbapenens. O objetivo deste trabalho foi detectar cepas de Pseudomonas aeruginosa resistentes ao imipenem e à ceftazidima, assim como identificar aquelas produtoras de MBLs. Foram estudadas 198 linhagens não-repetitivas isoladas de diversas amostras clínicas, hospitalares e comunitárias, identificadas bioquimica...

  3. Mutant prevention concentrations of four carbapenems against gram-negative rods.

    Science.gov (United States)

    Credito, Kim; Kosowska-Shick, Klaudia; Appelbaum, Peter C

    2010-06-01

    We tested the propensities of four carbapenems to select for resistant Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii mutants by determining the mutant prevention concentrations (MPCs) for 100 clinical strains with various ss-lactam phenotypes. Among the members of the Enterobacteriaceae family and A. baumannii strains, the MPC/MIC ratios were mostly 2 to 4. In contrast, for P. aeruginosa the MPC/MIC ratios were 4 to > or =16. The MPC/MIC ratios for beta-lactamase-positive K. pneumoniae and E. coli isolates were much higher (range, 4 to >16 microg/ml) than those for ss-lactamase-negative strains.

  4. [The development of antibiotics use in Danish food production].

    Science.gov (United States)

    Jensen, Vibeke Frøkjær

    2011-11-07

    In the Danish husbandry antimicrobial growth promoters were phased out 1994-2000 and the therapeutic use has been increasingly regulated. Hitherto, a minimum in therapeutic use was reached in 1997. The antimicrobial use in pigs increased by 44% during 2002-2009; a 12% decrease in second half of 2010 was likely due to the announcement of the "yellow card" regulation. From July 2010, a voluntary two years stop of cephalosporins use in pigs was realized, due to increasing occurrence of extended spectrum beta lactamase (ESBL) resistance in animal and meat isolates; highest levels of ESBL are observed in imported poultry.

  5. Effect of synthetic and natural phospholipids on N-acylphosphatidylethanolamine-hydrolyzing phospholipase D activity

    DEFF Research Database (Denmark)

    Petersen, Gitte; Pedersen, Anders H; Pickering, Darryl S

    2009-01-01

    N-Acylethanolamines (NAEs) constitute a family of endogenous bioactive lipids that includes arachidonoylethanolamide (anandamide), palmitoylethanolamide (PEA) and oleoylethanolamide (OEA). These lipids are formed from their respective N-acylated ethanolamine phospholipid (NAPE) precursor by the a...... analogues as well as selected phospholipids and beta-lactamase substrates were tested as potential modifiers of cloned human NAPE-PLD in an enzyme assay involving a (14)C-labeled diether-NAPE substrate. One hit was identified, namely 1,2-dihexanoyl-glycero-N-(3-(tetradecanoylamino...

  6. Aerobic and anaerobic bacteria in tonsils of children with recurrent tonsillitis.

    Science.gov (United States)

    Brook, I; Yocum, P; Friedman, E M

    1981-01-01

    Tonsils were obtained from 50 children suffering from recurrent tonsillitis. Patients' ages ranged from 2.5 to 17 years (mean 6 years); 29 were males and 21 females. The tonsils were sectioned in half after heat searing of the surface and the core material was cultured for aerobic and anaerobic microorganisms. Mixed aerobic and anaerobic flora was obtained in all patients, yielding an average of 7.8 isolates (4.1 anaerobes and 3.7 aerobes) per specimen. There were 207 anaerobes isolated. The predominant isolates were 101 Bacteroides sp (including 10 B fragilis group, and 47 B melaninogenicus group), 29 Fusobacterium sp, 34 Gram-positive anaerobic cocci (25 Peptococcus sp and 9 Peptostreptococcus sp) and 16 Veillonella sp. There were 185 aerobic isolates. The predominant isolates were 41 alpha-hemolytic streptococci, 24 Staphylococcus aureus, 19 beta-hemolytic streptococci (11 group A, 4 group B, and 2 each group C and F), 14 Haemophilus sp (including 12 H influenzae type B) and 5 H parainfluenzae. Beta-lactamase production was noted in 56 isolates recovered from 37 tonsils. These were all isolates of S aureus (24) and B fragilis (10), 15 of 47 B melaninogenicus (32%), 5 of the 12 B oralis (42%), and 2 of 12 H influenzae type B (17%). Our findings indicate the polymicrobial aerobic and anaerobic nature of deep tonsillar flora in children with recurrent tonsillitis, and demonstrate the presence of many beta-lactamase-producing organisms in 74% of the patients.

  7. The presence of plasmid-mediated resistance genes among uropathogenes isolated from diabetic and non-diabetic patients with chronic pyelonephritis

    Directory of Open Access Journals (Sweden)

    O.I. Chub

    2014-10-01

    Full Text Available Increased multidrug resistance of extended-spectrum beta-lactamases (ESBLs compromises the efficacy of treatment of urinary tract infections. The objective of this study is to determine the prevalence of ESBL-producing uropathogens from patients with chronic pyelonephritis (CP and to evaluate the risk factors of these types of infections. Screening for the presence of plasmid-mediated ESBL was performed by polymerase chain reaction. Out of 105 patients, 22 (20.9% revealed strains with resistance genes: 11 (36.7%, 11 (36.7% and 8 (26.7% were identified to carry bla(TEM, bla(SHV and bla(CTX-M beta-lactamase genes, respectively. We have demonstrated that prevalence of the resistance among patients with CP combined with type 2 DM was 31.3%, while among patients with CP without type 2 DM was 27.4%; however the difference between these groups was not significant. The main factors related with appearance of plasmid-mediated resistance genes were age range above 55 years, Chronic Kidney Disease stage ІІІ and ІV, in-patient treatment history, history of using antibiotics last year. Isolation and detection of ESBL-producing strains are essential fоr the sеlection оf the mоst effеctive antibiоtic for the empiric trеatment.

  8. Susceptibilities to carbapenems and presence of cphA gene on food-borne Aeromonas

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    Bibiana María Martín Talavera

    2006-07-01

    Full Text Available The purpose of this study was to determine the susceptibilities of food-borne Aeromonas to carbapenems, as well as to investigate the presence of a metallo carbapenemase-encoding gene, named cphA. Minimum Inhibitory Concentration (MIC was determined following NCCLS standards. All the tested microorganisms were susceptible to imipenem, meropenem and biapenem. However, a strong inoculum size effect on carbapenem MICs was observed for most of the strains. Six strains, out of seven, showed the presence of metallo--beta-lactamases but cphA gene was detected in only two strains of A. veronii bv. sobria.O objetivo deste estudo foi determinar a suscetibilidade de aeromonas de origem alimentar a carbapenems bem como investigar a presença de um gene codificante de metalocarbapenemase, denominado "cph A". A suscetibilidade in vitro foi determinada pelo metodo de diluição em agar. Todas as cepas foram suscetíveis a Imipenem, Meropenem e Biapenem. Porém foi observado um forte efeito de tamanho do inóculo sobre as CIM das carbapenems na maioria das cepas. A detecção de metalo-beta-lactamase foi realizada pelo metodo lodometrico. Seis cepas das sete testadas demostraron a presença da enzima. A presença do gene cphA foi determinada por PCR e foi detectada em duas cepas de A veronii bv. sobria.

  9. Ceftiofur sodium, a broad-spectrum cephalosporin: evaluation in vitro and in vivo in mice.

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    Yancey, R J; Kinney, M L; Roberts, B J; Goodenough, K R; Hamel, J C; Ford, C W

    1987-07-01

    Ceftiofur sodium, a broad-spectrum beta-lactamase-resistant cephalosporin, was evaluated in vitro and in vivo in mice. Ceftiofur is the sodium salt of (6R, 7R)-7[( 2-amino-4-thiazolyl)-Z- (methoxyimino)acetyl]amino)-3-[( (2-furanylcarbonyl)thio]methyl)-8-oxo-5- thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylate. Minimal inhibitory concentration values were obtained with 264 strains representing 9 genera and 17 species of bacterial pathogens from cattle, swine, sheep, horses, poultry, dogs, cats, and human beings. Ceftiofur was more active than was ampicillin against all strains tested including beta-lactamase-producing organisms. In mice with systemic infections, ceftiofur was more active than or equivalent to ampicillin, cephalothin, cefamandole, cloxacillin, cefoperazone, or pirlimycin. These protection tests included infections with Escherichia coli, Haemophilus pleuropneumoniae, H somnus, Pasteurella haemolytica, P multocida, Salmonella typhimurium, or Staphylococcus aureus. In infant mice with E coli-induced lethal diarrhea and in mice with S aureus and E coli-induced mastitis, ceftiofur was comparable or more active than was ampicillin.

  10. Antibiotic resistance in animals.

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    Barton, Mary D; Pratt, Rachael; Hart, Wendy S

    2003-01-01

    There is currently no systematic surveillance or monitoring of antibiotic resistance in Australian animals. Registration of antibiotics for use in animals is tightly controlled and has been very conservative. Fluoroquinolones have not been registered for use in food producing animals and other products have been removed from the market because of human health concerns. In the late 1970s, the Animal Health Committee coordinated a survey of resistance in Salmonella and Escherichia coli isolates from cattle, pigs and poultry and in bovine Staphylococcus aureus. Some additional information is available from published case reports. In samples collected prior to the withdrawal of avoparcin from the market, no vancomycin resistant Enterococcus faecium or Enterococcus faecalis were detected in samples collected from pigs, whereas some vanA enterococci, including E. faecium and E. faecalis, were found in chickens. No vanB enterococci were detected in either species. Virginiamycin resistance was common in both pig and poultry isolates. Multiple resistance was common in E. coli and salmonellae isolates. No fluoroquinolone resistance was found in salmonellae, E. coli or Campylobacter. Beta-lactamase production is common in isolates from bovine mastitis, but no methicillin resistance has been detected. However, methicillin resistance has been reported in canine isolates of Staphylococcus intermedius and extended spectrum beta-lactamase producing E. coli has been found in dogs.

  11. Resistencia de la bacteria Escherichiacoli por la beta-lactamasas

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    Armijos- Nieves, Bryan,

    2017-09-01

    Full Text Available E. coli bacterial infections are frequently present with multiple factors that make their treatment more expensive, in the health field at the national level. The objective of this work is to determine the influence of the beta-lactamase enzyme on E. coliresistance against antibiotics, recognizing the antibacterialsfrom the families of penicillins and cephalosporins inefficient in their treatment. Based on a documentary study of scientific articles on the subject under study, it is conclude that the presence of the enzyme beta-lactamase greatly influences the resistance developed by the bacterium E. coli towards antibiotics such as penicillins and cephalosporins first, second, third and fourth generation. In addition, it was determined that the antibiotics derived from penicillin, such as ampicillin and antibiotics of the cephalosporin family cefazolin, cephalothin, cefotaxime, cefepime and cefoptaxime by hydrolysis of these drugs, do not exert any antibacterial effect of resistant E. coli, representing a big problem that may increase the risk of patient mortality. It is recommended that, in case of suspected E. coli infection, an antibiogram be performed to detect the indicated treatment.

  12. High rates of multidrug resistance among uropathogenic Escherichia coli in children and analyses of ESBL producers from Nepal

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    Narayan Prasad Parajuli

    2017-01-01

    Full Text Available Abstract Background Emergence of Extended-spectrum beta-lactamase producing Escherichia coli causing urinary tract infections (UTI among pediatric patients is an increasing problem worldwide. However, very little is known about pediatric urinary tract infections and antimicrobial resistance trend from Nepal. This study was conducted to assess the current antibiotic resistance rate and ESBL production among uropathogenic Escherichia coli in pediatric patients of a tertiary care teaching hospital of Nepal. Methods A total of 5,484 urinary tract specimens from children suspected with UTI attending a teaching hospital of Nepal over a period of one year were processed for the isolation of bacterial pathogens and their antimicrobial susceptibility testing. Escherichia coli (n = 739, the predominant isolate in pediatric UTI, was further selected for the detection of ESBL-production by phenotypic combination disk diffusion test. Results Incidence of urinary tract infection among pediatric patients was found to be 19.68% and E coli (68.4% was leading pathogen involved. Out of 739 E coli isolates, 64.9% were multidrug resistant (MDR and 5% were extensively drug resistant (XDR. Extended spectrum beta lactamase (ESBL was detected in 288 (38.9% of the E coli isolates. Conclusion Alarming rate of drug resistance among pediatric uropathogens and high rate of ESBL-producing E. coli was observed. It is extremely necessary to routinely investigate the drug resistance among all isolates and formulate strict antibiotics prescription policy in our country.

  13. [Consensus for antimicrobial susceptibility testing for Enterobacteriaceae. Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology].

    Science.gov (United States)

    Famiglietti, A; Quinteros, M; Vázquez, M; Marín, M; Nicola, F; Radice, M; Galas, M; Pasterán, F; Bantar, C; Casellas, J M; Kovensky Pupko, J; Couto, E; Goldberg, M; Lopardo, H; Gutkind, G; Soloaga, R

    2005-01-01

    Taking into account previous recommendations from the National Committee for Clinical Laboratory Standards (NCCLS), the Antimicrobial Committee, Sociedad Argentina de Bacteriología Clínica (SADEBAC), Asociación Argentina de Microbiología (AAM), and the experience from its members and some invited microbiologists, a consensus was obtained for antimicrobial susceptibility testing and interpretation in most frequent enterobacterial species isolated from clinical samples in our region. This document describes the natural antimicrobial resistance of some Enterobacteriaceae family members, including the resistance profiles due to their own chromosomal encoded beta-lactamases. A list of the antimicrobial agents that should be tested, their position on the agar plates, in order to detect the most frequent antimicrobial resistance mechanisms, and considerations on which antimicrobial agents should be reported regarding to the infection site and patient characteristics are included. Also, a description on appropriate phenotypic screening and confirmatory test for detection of prevalent extended spectrum beta-lactamases in our region are presented. Finally, a summary on frequent antimicrobial susceptibility profiles and their probably associated resistance mechanisms, and some infrequent antimicrobial resistance profiles that deserve confirmation are outlined.

  14. Treatment Options for Infections Caused by Carbapenem-resistant Enterobacteriaceae: Can We Apply “Precision Medicine” to Antimicrobial Chemotherapy?

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    Perez, Federico; El Chakhtoura, Nadim G.; Papp-Wallace, Krisztina; Wilson, Brigid M; Bonomo, Robert A.

    2016-01-01

    Introduction For the past three decades, carbapenems played a central role in our antibiotic armamentarium, trusted to effectively treat infections caused by drug-resistant bacteria. The utility of this class of antibiotics has been compromised by the emergence of resistance especially among Enterobacteriaceae. Areas covered We review the current mainstays of pharmacotherapy against infections caused by carbapenem-resistant Enterobacteriaceae (CRE) including tigecycline, aminoglycosides, and rediscovered 'old' antibiotics such as fosfomycin and polymyxins, and discuss their efficacy and potential toxicity. We also summarize the clinical experience treating CRE infections with antibiotic combination therapy. Finally, we review ceftazidime/avibactam and imipenem/relebactam, a new generation of beta-lactamase inhibitors, which may offer alternatives to treat CRE infections. We critically evaluate the published literature, identify relevant clinical trials and review documents submitted to the United States Food and Drug Administration. Expert Opinion It is essential to define the molecular mechanisms of resistance and to apply insights about pharmacodynamic and pharmacokinetic properties of antibiotics, in order to maximize the impact of old and new therapeutic approaches against infections caused by CRE. A concerted effort is needed to carry out high-quality clinical trials that: i) establish the superiority of combination therapy vs. monotherapy; ii) confirm the role of novel beta-lactam/beta-lactamase inhibitor combinations as therapy against KPC- and OXA-48 producing Enterobacteriaceae; and, iii) evaluate new antibiotics active against CRE as they are introduced into the clinic. PMID:26799840

  15. Multidrug-resistant Enterobacteriaceae from indoor air of an urban wastewater treatment plant.

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    Teixeira, Juliana V; Cecílio, Pedro; Gonçalves, Daniela; Vilar, Vítor J P; Pinto, Eugénia; Ferreira, Helena N

    2016-07-01

    Wastewater treatment plants (WWTPs) have been recognized as sources of bioaerosols that may act as vehicles for dissemination of pathogens and multidrug-resistant (MDR) bacteria. The occurrence of MDR Enterobacteriaceae in indoor air of an urban WWTP was investigated. A possible airborne contamination with extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing Enterobacteriaceae was also explored. Fourteen of 39 Enterobacteriaceae isolates were MDR. These isolates were found at all sampling sites, mainly at the secondary sedimentation settings. The highest levels of resistance were detected in three different species: Enterobacter cloacae, Escherichia coli, and Citrobacter freundii. Furthermore, one of the airborne E. coli isolates was phenotypically characterized as an ESBL producer. Additionally, five isolates showed non-susceptibility to at least one carbapenem tested. The presence of genes encoding relevant beta-lactamase types in these ESBL-producing and carbapenem-resistant Enterobacteriaceae isolates was investigated by PCR. Results showed amplification for bla CTX-M and bla OXA. These findings are relevant both in terms of occupational/public health and of environmental dissemination of MDR bacteria.

  16. Evaluation of antibacterial activities of flomoxef against ESBL producing Enterobacteriaceae analyzed by Monte Carlo simulation.

    Science.gov (United States)

    Ito, Akinobu; Tatsumi, Yumiko Matsuo; Wajima, Toshihiro; Nakamura, Rio; Tsuji, Masakatsu

    2013-04-01

    The growing number of infection caused by extended-spectrum beta-lactamase (ESBL) producing pathogens has prompted a more rational use of available antibiotics because of the paucity of new, effective agents. Flomoxef (FMOX) is one of the beta-lactam antibiotic which is stable against beta-lactamase. In this study, the antibacterial activity of FMOX was investigated, and Monte Carlo Simulation was conducted to determine the appropriate dosing regimens of FMOX based on the probability of target attainment (TA%) at the critical drug exposure metric of time that drug concentrations remain above 40% (showing bacteriostatic effect) or 70% (showing bactericidal effect) of time during which plasma concentration above minimum inhibitory concentration (MIC) of the drug (T(>MIC)) against the ESBL producing Enterobacteriaceae. The effective regimens to achieve 80% of TA% at 70% of T(>MIC) were 1 g every 8 hours with 2-4 hours infusion, and 1 g every 6 hours with 1-4 hours infusion. Moreover, all the tested regimens were effective to achieve 80% of TA% at 40% of T(>MIC). These results of pharmacokinetics/ pharmacodynamics (PK/PD) modeling showed the potential efficacy of FMOX against bacterial infections caused by ESBL producing Enterobacteriaceae.

  17. Risk factors for extended-spectrum b-lactamases-producing Escherichia coli urinary tract infections in a tertiary hospital

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    María Dolores Alcántar-Curiel

    2015-09-01

    Full Text Available Objective. To assess the risks factors for urinary tract infections (UTIs caused by Extended-Spectrum Beta-Lactamases (ESBLs-producing E. coli and the molecular characterization of ESBLs. Materials and methods. A case-control study was performed to identify risk factors in consecutively recruited patients with UTIs caused by ESBLs or non-ESBLs-producing E. coli in a tertiary hospital in Mexico. Results. ESBLs-producing E. coli were isolated from 22/70 (31% patients with E. coli UTIs over a three month period. All isolates were resistant to cephalosporins and quinolones but susceptible to carbapenems, amikacin and nitrofurantoin. Prior antibiotic treatment with more than two antibiotic families (OR=6.86; 95%CI 1.06-157.70; p=0.028, recurrent symptomatic UTIs (OR=5.60; 95%CI 1.88-17.87; p=0.001 and previous hospitalization (OR=5.06; 95%CI 1.64-17.69;p=0.002 were significant risk factors. Sixteen isolates harbored the beta-lactamase (blaCTX-M-15 gene and five the blaTEM-1 gene. Conclusions. One of every three patients presented UTIs with ESBLs-producing beta-lactams and fluoroquinolone resistant E. coli. Risk factors and resistance patterns must be taken into account for developing antibiotic use policies in these settings

  18. Sedation, sucralfate, and antibiotic use are potential means for protection against early-onset ventilator-associated pneumonia.

    Science.gov (United States)

    Bornstain, C; Azoulay, E; De Lassence, A; Cohen, Y; Costa, M A; Mourvillier, B; Descorps-Declere, A; Garrouste-Orgeas, M; Thuong, M; Schlemmer, B; Timsit, J-F

    2004-05-15

    To examine risk factors for early-onset ventilator-associated pneumonia (EOP) in patients requiring mechanical ventilation (MV), we performed a prospective cohort study that included 747 patients. Pneumonia was defined as a positive result for a protected quantitative distal sample. EOP was defined as pneumonia that occurred from day 3 to day 7 of MV. Eighty patients (10.7%) experienced EOP. Independent predictors of EOP were male sex (odds ratio [OR], 2.06; 95% confidence interval [CI], 1.18-3.63), actual Glasgow Coma Scale value of 6-13 (OR, 1.95; 95% CI, 1.2-3.18), high Logistic Organ Dysfunction score at day 2 (OR, 1.12 per point; 95% CI, 1.02-1.23), unplanned extubation (OR, 3.19; 95% CI, 1.28-7.92), and sucralfate use (OR, 1.81; 95% CI, 1.01-3.26). Protection occurred with use of aminoglycosides (OR, 0.36; 95% CI, 0.17-0.76), beta -lactams and/or beta -lactamase inhibitors (OR, 0.47; 95% CI, 0.28-0.83), or third-generation cephalosporins (OR, 0.33; 95% CI, 0.16-0.74). Sucralfate use and unplanned extubation are independent risk factors for EOP. Use of aminoglycosides, beta-lactams/ beta-lactamase inhibitors, or third-generation cephalosporins protects against EOP.

  19. Emergence of KPC-producing Klebsiella pneumoniae in Italy

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    Bossa Maria C

    2010-02-01

    Full Text Available Abstract Background The emergence of KPC-producing K. pneumoniae has now become a global concern. KPC beta-lactamases are plasmid-borne and, like extended spectrum beta lactamases (ESBLs, can accumulate and transfer resistance determinants to other classes of antibiotics. Therefore, infection control guidelines on early identification and control of the spread of organisms carrying these resistant determinants are needed. Findings Klebsiella pneumoniae carbapenemase (KPC was detected in two isolates of carbapenem-resistant K. pneumoniae obtained from patients at an Italian teaching hospital. The first strain was isolated from a culture drawn from a central venous device (CVC in a patient with Crohn's disease who was admitted to a gastroenterology ward. The second was isolated from a urine sample collected from an indwelling urinary catheter in an intensive care unit (ICU patient with a subdural haematoma. The patients had not travelled abroad. Both isolates were resistant to all β-lactams and were susceptible to imipenem and meropenem but resistant to ertapenem. Isolates also showed resistance to other classes of non-β-lactam antibiotics, such as quinolones, aminoglycosides (with the exception for amikacin, trimethoprim-sulfamethoxazole (TMP-SMX and nitrofurantoin. They were determined to contain the plasmid encoding the carbapenemase gene bla-KPC and were also positive in the Hodge test. Conclusions This is the second report of KPC-producing isolates in Italy, but the first concerning KPC type 2 gene, and it may have important implications for controlling the transmission of microorganisms resistant to antibiotics.

  20. In vitro selection of resistance in haemophilus influenzae by 4 quinolones and 5 beta-lactams.

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    Clark, Catherine; Kosowska, Klaudia; Bozdogan, Bülent; Credito, Kim; Dewasse, Bonifacio; McGhee, Pamela; Jacobs, Michael R; Appelbaum, Peter C

    2004-05-01

    We tested abilities of ciprofloxacin, levofloxacin, gatifloxacin, moxifloxacin, amoxicillin, amoxicillin/clavulanate, cefixime, cefpodoxime, and cefdinir to select resistant mutants in 5 beta-lactamase positive and 5 beta-lactamase negative Haemophilus influenzae strains by single and multistep methodology. In multistep tests, amoxicillin, amoxicillin/clavulanate and cefpodoxime exposure did not cause >4-fold minimum inhibitory concentration (MIC) increase after 50 days. One mutant selected by cefdinir had one amino acid substitution (Gly490Glu) in PBP3 and became resistant to cefdinir. Cefixime exposure caused 8-fold MIC-increase in 1 strain with TEM but the mutant remained cefixime susceptible and had no alteration in PBP3 or TEM. Among 10 strains tested, ciprofloxacin, moxifloxacin, gatifloxacin, levofloxacin caused >4-fold MIC increase in 6, 6, 5, and 2 strain, respectively. Despite the increases in quinolone MICs, none of the mutants became resistant to quinolones by established criteria. Quinolone selected mutants had quindone resistance-determining region (QRDR) alterations in GyrA, GyrB, ParC, ParE. Four quinolone mutants had no QRDR alterations. Among beta-lactams cefdinir and cefixime selected one mutant each with higher MICs however amoxicillin, amoxicillin/clavulanate, and cefpodoxime exposure did not select resistant mutants.

  1. Doripenem: an expected arrival in the treatment of infections caused by multidrug-resistant Gram-negative pathogens.

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    Poulakou, Garyphallia; Giamarellou, Helen

    2008-05-01

    Potent new drugs against multidrug-resistant Gram-negative bacteria, namely Pseudomonas aeruginosa and Acinetobacter spp. and pan-drug-resistant Klebsiella pneumoniae, which constitute an increasing medical threat, are almost absent from the future pharmaceutical pipeline. This drug evaluation focuses on the position of doripenem, a novel forthcoming carbapenem. Mechanisms of resistance and new drugs with anti-Gram-negative activity are also briefly reviewed. Literature search was performed for new carbapenems, new antibiotics, doripenem, metallo-beta-lactamase inhibitors, multidrug-resistant pathogens, antipseudomonal antibiotics and multidrug-resistant epidemiology. Doripenem possesses a broad spectrum of activity against Gram-negative bacteria, similar to that of meropenem, while retaining the spectrum of imipenem against Gram-positive pathogens. Against P. aeruginosa, doripenem exhibits rapid bactericidal activity with 2 - 4-fold lower MIC values, compared to meropenem. Exploitation of pharmacokinetic/pharmacodynamic applications could offer a treatment opportunity against strains exhibiting borderline resistance to doripenem. Stability against numerous beta-lactamases, low adverse event potential and more potent in vitro antibacterial activity against P. aeruginosa and A. baumanni compared to the existing carbapenems, are its principal features.

  2. In vitro susceptibility pattern of extended spectrum ?-lactamase producing gram negative bacilli against tetracyclines

    International Nuclear Information System (INIS)

    Gill, M.M.

    2015-01-01

    Extended Spectrum beta-lactamases (ESBLs) are emerging as common nosocomial pathogens and important cause of mortality and morbidity, if not treated properly. The need of the hour is to find effective treatment options for dealing with ESBL producing organisms. This study was aimed to evaluate in vitro susceptibility pattern of extended spectrum beta-lactamase producers against tetracyclines. Methods: This descriptive cross-sectional study was carried out in the department of Microbiology, Army Medical College, Rawalpindi, National University of Sciences and Technology over a period of 6 months. Seventy eight non-duplicate isolates were included in the study. ESBL detection was done using Jarlier et al method. In vitro susceptibility of tetracyclines like tetracycline, doxycycline, minocycline and tigecycline was then tested using Modified Kirby Bauer disc diffusion method. The zones of inhibition were measured after completion of incubation period and interpreted as per CLSI and FDA guidelines. Results: Approximately 56.4% of the isolates were Escherichia coli, 28.2% were Klebsiella pneumoniae, 10.26% were Enterobacter species, and 2.6% were each Klebsiella oxytoca and Acinetobacter species. ESBLs were found to be most sensitive to tigecycline, intermediate in susceptibility to minocycline while least sensitive to doxycycline and tetracycline. Conclusion: Among tetracyclines, tigecycline has best in vitro susceptibility against ESBL producing Gram negative rods. (author)

  3. Dual beta-lactam therapy for serious Gram-negative infections: is it time to revisit?

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    Rahme, Christine; Butterfield, Jill M; Nicasio, Anthony M; Lodise, Thomas P

    2014-12-01

    We are rapidly approaching a crisis in antibiotic resistance, particularly among Gram-negative pathogens. This, coupled with the slow development of novel antimicrobial agents, underscores the exigency of redeploying existing antimicrobial agents in innovative ways. One therapeutic approach that was heavily studied in the 1980s but abandoned over time is dual beta-lactam therapy. This article reviews the evidence for combination beta-lactam therapy. Overall, in vitro, animal and clinical data are positive and suggest that beta-lactam combinations produce a synergistic effect against Gram-negative pathogens that rivals that of beta-lactam-aminoglycoside or beta-lactam-fluoroquinolone combination therapy. Although the precise mechanism of improved activity is not completely understood, it is likely attributable to an enhanced affinity to the diverse penicillin-binding proteins found among Gram negatives. The collective data indicate that dual beta-lactam therapy should be revisited for serious Gram-negative infections, especially in light of the near availability of potent beta-lactamase inhibitors, which neutralize the effect of problematic beta-lactamases. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. In vitro activity of tigecycline and comparator agents against a global collection of Gram-negative and Gram-positive organisms: tigecycline Evaluation and Surveillance Trial 2004 to 2007.

    Science.gov (United States)

    Garrison, Mark W; Mutters, Reinier; Dowzicky, Michael J

    2009-11-01

    The Tigecycline Evaluation and Surveillance Trial began in 2004 to monitor the in vitro activity of tigecycline and comparator agents against a global collection of Gram-negative and Gram-positive pathogens. Against Gram negatives (n = 63 699), tigecycline MIC(90)'s ranged from 0.25 to 2 mg/L for Escherichia coli, Haemophilus influenzae, Acinetobacter baumannii, Klebsiella oxytoca, Enterobacter cloacae, Klebsiella pneumoniae, and Serratia marcescens (but was > or =32 for Pseudomonas aeruginosa). Against Gram-positive organisms (n = 32 218), tigecycline MIC(90)'s were between 0.06 and 0.25 mg/L for Streptococcus pneumoniae, Enterococcus faecium, Streptococcus agalactiae, Staphylococcus aureus, and Enterococcus faecalis. The in vitro activity of tigecycline was maintained against resistant phenotypes, including multidrug-resistant A. baumannii (9.2% of isolates), extended-spectrum beta-lactamase-producing E. coli (7.0%) and K. pneumoniae (14.0%), beta-lactamase-producing H. influenzae (22.2%), methicillin-resistant S. aureus (44.5%), vancomycin-resistant E. faecium (45.9%) and E. faecalis (2.8%), and penicillin-resistant S. pneumoniae (13.8%). Tigecycline represents a welcome addition to the armamentarium against difficult to treat organisms.

  5. In vitro susceptibility testing to antimicrobial agents of urinary tract infection bacteria in women: a 5-year retrospective study

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    Laura Melgarejo

    2017-06-01

    Full Text Available Introduction: The growing resistance rates of urinary pathogens represent a serious problem. The aim of this study was to analyze the etiology of community-acquired urinary tract infections, their first-line antimicrobial resistance and the presence of extended-spectrum beta-lactamases (ESBL in gram negative bacilli. Methods: The study was conducted between January 2011 and December 2015 using data from the Microbiology Laboratory at the teaching hospital Hospital de Clínicas, which belongs to the National University of Asunción. Results: A total of 1957 urinary pathogens were found in women. Escherichia coli was the most commonly isolated bacterium (57%, followed by Klebsiella pneumoniae (11% and Streptococcus agalactiae (2%, Staphylococcus saprophyticus (2% and Proteus mirabilis (2%. The resistance rates of Escherichia coli were the following: to trimetoprim-sulfametoxazol, 43%; to ciprofloxacin, 32%; to ampicilin/sulbactam, 32%; to cefotaxime, 13 %; to piperacillin/tazobactam, 8%; nitrofurantoin, 2%, whereas it did not show resistance to meropenem during this period. Extended-spectrum beta-lactamases were produced by 11% of the E. coli isolates and 30% of the Klebsiella pneumoniae isolates. Conclusions: The resistance and cross-resistance rates found in this study pose a serious problem which compels the continuous assessment of the empirical therapy for urinary tract infections at this hospital.

  6. [Mechanisms of endogenous drug resistance acquisition by spontaneous chromosomal gene mutation].

    Science.gov (United States)

    Fukuda, H; Hiramatsu, K

    1997-05-01

    Endogenous resistance in bacteria is caused by a change or loss of function and generally genetically recessive. However, this type of resistance acquisition are now prevalent in clinical setting. Chromosomal genes that afford endogenous resistance are the genes correlated with the target of the drug, the drug inactivating enzymes, and permeability of the molecules including the antibacterial agents. Endogenous alteration of the drug target are mediated by the spontaneous mutation of their structural gene. This mutation provides much lower affinity of the drugs for the target. Gene expression of the inactivating enzymes, such as class C beta-lactamase, is generally regulated by regulatory genes. Spontaneous mutations in the regulatory genes cause constitutive enzyme production and provides the resistant to the agent which is usually stable for such enzymes. Spontaneous mutation in the structural gene gives the enzyme extra-spectrum substrate specificity, like ESBL (Extra-Spectrum-beta-Lactamase). Expression of structural genes encoding the permeability systems are also regulated by some regulatory genes. The spontaneous mutation of the regulatory genes reduce an amount of porin protein. This mutation causes much lower influx of the drug in the cell. Spontaneous mutation in promoter region of the structural gene of efflux protein was observed. This mutation raised the gene transcription and overproduced efflux protein. This protein progresses the drug efflux from the cell.

  7. Detection of Methicillin Resistant Staphylococcus aureus and Determination of Minimum Inhibitory Concentration of Vancomycin for Staphylococcus aureus Isolated from Pus/Wound Swab Samples of the Patients Attending a Tertiary Care Hospital in Kathmandu, Nepal

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    Raghabendra Adhikari

    2017-01-01

    Full Text Available The present study was conducted to evaluate the performance of cefoxitin disc diffusion method and oxacillin broth microdilution method for detection of methicillin resistant S. aureus (MRSA, taking presence of mecA gene as reference. In addition, inducible clindamycin resistance and beta-lactamase production were studied and minimum inhibitory concentration (MIC of vancomycin for S. aureus isolates was determined. A total of 711 nonrepeated pus/wound swab samples from different anatomic locations were included in the study. The Staphylococcus aureus was identified on the basis of colony morphology, Gram’s stain, and biochemical tests. A total of 110 (15.47% S. aureus isolates were recovered, of which 39 (35.50% isolates were identified as MRSA by cefoxitin disc diffusion method. By oxacillin broth microdilution method, 31.82% of the Staphylococcus aureus isolates were found to be MRSA. However, mecA gene was present in only 29.1% of the isolates. Further, beta-lactamase production was observed in 71.82% of the isolates, while inducible clindamycin resistance was found in 10% of S. aureus isolates. The MIC value of vancomycin for S. aureus ranged from 0.016 μg/mL to 1 μg/mL. On the basis of the absolute sensitivity (100%, both phenotypic methods could be employed for routine diagnosis of MRSA in clinical microbiology laboratory; however cefoxitin disc diffusion could be preferred over MIC method considering time and labour factor.

  8. Machine Learning Leveraging Genomes from Metagenomes Identifies Influential Antibiotic Resistance Genes in the Infant Gut Microbiome

    Science.gov (United States)

    Olm, Matthew R.; Morowitz, Michael J.

    2018-01-01

    ABSTRACT Antibiotic resistance in pathogens is extensively studied, and yet little is known about how antibiotic resistance genes of typical gut bacteria influence microbiome dynamics. Here, we leveraged genomes from metagenomes to investigate how genes of the premature infant gut resistome correspond to the ability of bacteria to survive under certain environmental and clinical conditions. We found that formula feeding impacts the resistome. Random forest models corroborated by statistical tests revealed that the gut resistome of formula-fed infants is enriched in class D beta-lactamase genes. Interestingly, Clostridium difficile strains harboring this gene are at higher abundance in formula-fed infants than C. difficile strains lacking this gene. Organisms with genes for major facilitator superfamily drug efflux pumps have higher replication rates under all conditions, even in the absence of antibiotic therapy. Using a machine learning approach, we identified genes that are predictive of an organism’s direction of change in relative abundance after administration of vancomycin and cephalosporin antibiotics. The most accurate results were obtained by reducing annotated genomic data to five principal components classified by boosted decision trees. Among the genes involved in predicting whether an organism increased in relative abundance after treatment are those that encode subclass B2 beta-lactamases and transcriptional regulators of vancomycin resistance. This demonstrates that machine learning applied to genome-resolved metagenomics data can identify key genes for survival after antibiotics treatment and predict how organisms in the gut microbiome will respond to antibiotic administration. IMPORTANCE The process of reconstructing genomes from environmental sequence data (genome-resolved metagenomics) allows unique insight into microbial systems. We apply this technique to investigate how the antibiotic resistance genes of bacteria affect their ability to

  9. Occurrence and characteristics of extended-spectrum β-lactamase (ESBL producing Enterobacteriaceae in food producing animals, minced meat and raw milk

    Directory of Open Access Journals (Sweden)

    Geser Nadine

    2012-03-01

    Full Text Available Abstract Background The impact of food animals as a possible reservoir for extended-spectrum beta-lactamase (ESBL producing Enterobacteriaceae, and the dissemination of such strains into the food production chain need to be assessed. In this study 334 fecal samples from pigs, cattle, chicken and sheep were investigated at slaughter. Additionally, 100 raw milk samples, representing bulk tank milk of 100 different dairy farms, 104 minced meat (pork and beef samples and 67 E. coli isolates from cattle E. coli mastitis were analyzed. Results As many as 15.3% of the porcine, 13.7% of the bovine, 8.6% of the sheep and 63.4% of the chicken fecal samples yielded ESBL producers after an enrichment step. In contrast, none of the minced meat, none of the bulk tank milk samples and only one of the mastitis milk samples contained ESBL producing strains. Of the total of 91 isolates, 89 were E. coli, one was Citrobacter youngae and one was Enterobacter cloacae. PCR analysis revealed that 78 isolates (85.7% produced CTX-M group 1 ESBLs while six isolates (6.6% produced CTX-M group 9 enzymes. Five detected ESBLs (5.5% belonged to the SHV group and 2 isolates (2.2% contained a TEM-type enzyme. A total of 27 CTX-M producers were additionally PCR-positive for TEM-beta-lactamase. The ESBL-encoding genes of 53 isolates were sequenced of which 34 produced CTX-M-1, 6 produced CTX-M-14, 5 produced CTX-M-15 and also 5 produced SHV-12. Two isolates produced TEM-52 and one isolate expressed a novel CTX-M group 1 ESBL, CTX-M-117. One isolate--aside from a CTX-M ESBL-- contained an additional novel TEM-type broad-spectrum beta-lactamase, TEM-186. Conclusions The relatively high rates of ESBL producers in food animals and the high genetic diversity among these isolates are worrisome and indicate an established reservoir in farm animals.

  10. Detection of P. aeruginosa harboring bla CTX-M-2, bla GES-1 and bla GES-5, bla IMP-1 and bla SPM-1 causing infections in Brazilian tertiary-care hospital

    Science.gov (United States)

    2012-01-01

    Background Nosocomial infections caused by Pseudomonas aeruginosa presenting resistance to beta-lactam drugs are one of the most challenging targets for antimicrobial therapy, leading to substantial increase in mortality rates in hospitals worldwide. In this context, P. aeruginosa harboring acquired mechanisms of resistance, such as production of metallo-beta-lactamase (MBLs) and extended-spectrum beta-lactamases (ESBLs) have the highest clinical impact. Hence, this study was designed to investigate the presence of genes codifying for MBLs and ESBLs among carbapenem resistant P. aeruginosa isolated in a Brazilian 720-bed teaching tertiary care hospital. Methods Fifty-six carbapenem-resistant P. aeruginosa strains were evaluated for the presence of MBL and ESBL genes. Strains presenting MBL and/or ESBL genes were submitted to pulsed-field gel electrophoresis for genetic similarity evaluation. Results Despite the carbapenem resistance, genes for MBLs (blaSPM-1 or blaIMP-1) were detected in only 26.7% of isolates. Genes encoding ESBLs were detected in 23.2% of isolates. The blaCTX-M-2 was the most prevalent ESBL gene (19.6%), followed by blaGES-1 and blaGES-5 detected in one isolate each. In all isolates presenting MBL phenotype by double-disc synergy test (DDST), the blaSPM-1 or blaIMP-1 genes were detected. In addition, blaIMP-1 was also detected in three isolates which did not display any MBL phenotype. These isolates also presented the blaCTX-M-2 gene. The co-existence of blaCTX-M-2 with blaIMP-1 is presently reported for the first time, as like as co-existence of blaGES-1 with blaIMP-1. Conclusions In this study MBLs production was not the major mechanism of resistance to carbapenems, suggesting the occurrence of multidrug efflux pumps, reduction in porin channels and production of other beta-lactamases. The detection of blaCTX-M-2,blaGES-1 and blaGES-5 reflects the recent emergence of ESBLs among antimicrobial resistant P. aeruginosa and the extraordinary

  11. Detection of P. aeruginosa harboring bla CTX-M-2, bla GES-1 and bla GES-5, bla IMP-1 and bla SPM-1 causing infections in Brazilian tertiary-care hospital

    Directory of Open Access Journals (Sweden)

    Polotto Milena

    2012-08-01

    Full Text Available Abstract Background Nosocomial infections caused by Pseudomonas aeruginosa presenting resistance to beta-lactam drugs are one of the most challenging targets for antimicrobial therapy, leading to substantial increase in mortality rates in hospitals worldwide. In this context, P. aeruginosa harboring acquired mechanisms of resistance, such as production of metallo-beta-lactamase (MBLs and extended-spectrum beta-lactamases (ESBLs have the highest clinical impact. Hence, this study was designed to investigate the presence of genes codifying for MBLs and ESBLs among carbapenem resistant P. aeruginosa isolated in a Brazilian 720-bed teaching tertiary care hospital. Methods Fifty-six carbapenem-resistant P. aeruginosa strains were evaluated for the presence of MBL and ESBL genes. Strains presenting MBL and/or ESBL genes were submitted to pulsed-field gel electrophoresis for genetic similarity evaluation. Results Despite the carbapenem resistance, genes for MBLs (blaSPM-1 or blaIMP-1 were detected in only 26.7% of isolates. Genes encoding ESBLs were detected in 23.2% of isolates. The blaCTX-M-2 was the most prevalent ESBL gene (19.6%, followed by blaGES-1 and blaGES-5 detected in one isolate each. In all isolates presenting MBL phenotype by double-disc synergy test (DDST, the blaSPM-1 or blaIMP-1 genes were detected. In addition, blaIMP-1 was also detected in three isolates which did not display any MBL phenotype. These isolates also presented the blaCTX-M-2 gene. The co-existence of blaCTX-M-2 with blaIMP-1 is presently reported for the first time, as like as co-existence of blaGES-1 with blaIMP-1. Conclusions In this study MBLs production was not the major mechanism of resistance to carbapenems, suggesting the occurrence of multidrug efflux pumps, reduction in porin channels and production of other beta-lactamases. The detection of blaCTX-M-2,blaGES-1 and blaGES-5 reflects the recent emergence of ESBLs among antimicrobial resistant P. aeruginosa and

  12. Subacute Bacterial Endocarditis Caused by Cardiobacterium hominis: A Case Report

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    Davie Wong

    2015-01-01

    Full Text Available Cardiobacterium hominis, a member of the HACEK group of organisms, is an uncommon but important cause of subacute bacterial endocarditis. First-line therapy is a third-generation cephalosporin due to rare beta-lactamase production. The authors report a case involving endovascular infection due to C hominis that initially tested resistant to third-generation cephalosporins using an antibiotic gradient strip susceptibility method (nitrocephin negative, but later proved to be susceptible using broth microdilution reference methods (a ‘major’ error. There are limited studies to guide susceptibility testing and interpretive breakpoints for C hominis in the medical literature, and the present case illustrates some of the issues that may arise when performing susceptibility testing for fastidious organisms in the clinical microbiology laboratory.

  13. Eficácia e segurança de Sultamicilina (Ampicilina/Sulbactam e Amoxacilina/Clavulanato no tratamento das infecções de via aéreas superiores em adultos: um estudo multicêntrico, aberto e randomizado Efficacy and safety of Sultamicillin (Ampicillin/Sulbactan and Amoxicillin/Clavulanic Acid in the treatment of upper respiratory tract infections in adults: an open-label, multicentric, randomized trial

    Directory of Open Access Journals (Sweden)

    João Batista Ferreira

    2006-02-01

    Full Text Available As IVAS em crianças e adultos são os motivos mais freqüentes de consulta médica e os que mais demandam o uso de antibióticos. A crescente resistência bacteriana causada pela produção das beta-lactamases constitui um dos mais sérios problemas atuais. A Sultamicilina é uma pró-droga dupla da ampicilina e do sulbactam, um potente inibidor de beta-lactamases que pode fazer frente a estas dificuldades. OBJETIVO: avaliar a eficácia, segurança e tolerabilidade da Ampicilina/Sulbactan comparada à Amoxacilina/Ácido Clavulânico no tratamento de IVAS, em adultos. METODOLOGIA: 102 pacientes com diagnóstico de IVAS foram randomizados em dois grupos recebendo Ampicilina/Sulbactan ou Amoxacilina/Clavulanato por 10 dias. Foram avaliados 10 e 30 dias após para análise da resposta terapêutica. RESULTADOS: Não houve diferença entre os grupos com relação à proporção de pacientes curados ao final do tratamento (visita 2 ou do estudo (visita 3. No grupo que recebeu Amoxacilina/Clavulanato, as proporções de cura foram de 61.7% e 93.2% nas visitas 2 e 3, comparadas a 64.4% e 97.4%, respectivamente, no grupo que recebeu Ampicilina/Sulbactan. A proporção de pacientes que experimentou pelo menos um evento adverso foi semelhante nos dois grupos (p = 0.940. A diarréia foi significativamente mais freqüente no grupo Amoxacilina-Clavulanato (70.6% do que no grupo Ampicilina/Sulbactan (29.4%, (p=0.0164. CONCLUSÕES: A Ampicilina/Sulbactan é tão segura e eficaz quanto a Amoxacilina/Clavulanato no tratamento empírico de IVAS em adultos. A ocorrência significativamente menor de quadros de diarréia no grupo recebendo Ampicilina/Sulbactan necessita confirmação em estudos posteriores.Upper respiratory tract infections are the most common causes of medical visits in children and adults, demanding massive use of antibiotics. Bacterial resistance caused by beta-lactamase is one of the most serious problems in this matter. Sultamicillin, a double

  14. Clinicomicrobiological analysis of patients with cholangitis

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    S M Shenoy

    2014-01-01

    Full Text Available Acute cholangitis is inflammation of biliary ductal system from infection with an associated biliary obstruction. This retrospective study was done to determine the factors responsible for cholangitis and the microbiological profile of the bile in patients with cholangitis. In the study involving 348 patients, 36.4% had associated malignancy. A total of 54% of the bile samples were positive for aerobic culture. Nearly 66-73% of the Escherichia coli and Klebsiella isolates were Extended spectrum beta lactamases (ESBL producers. Two isolates of Candida spps were also obtained. Polymicrobial infection was seen in 31.5% of the culture positive cases. Ideal antibiotics in case of cholangitis would be those which are excreted in the bile such as third-generation cephalosporins, ureidopenicillins, carbapenems and fluoroquinolones to combat resistance and polymicrobial aetiology. Anti-fungal drugs may also be necessary if the patient is not responding to biliary decompression and antibacterial agents to prevent fungaemia.

  15. Comparative Efficacy of Penicillin and Doxycycline in Gonococcal Urethritis

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    Vinod K Sharma

    1987-01-01

    Full Text Available Ninety two episodes of gonococcal urethritis treated with one of the following regimens viz: (A 3 m. u. of fortified procaine penicillin made by adding 1 m.u. of crystalline penicillin to 2 m.u. of fortified procaine penicillin intramuscularly with one gram of probenecid orally, (B 3 m.u. of above fortified procaine penicillin intramuscularly alone, and (C 400 mg single oral dose of doxycycline produced success rates of 95,76.2 and 66.7% respectively. Post gonococcal urethritis was detected in 37% patients. Thirty four (45.94% of the 74 isolates of N. gonorrhoeae were relatively resistant (MIC 0. 12 units ml to penicillin. None of the 74 Neisseria gonorrhoeac strains was beta lactamase producing.

  16. [Lower urinary tract infections in men. Urethritis, cystitis and prostatitis].

    Science.gov (United States)

    Zanella, Marie-Céline; Schoofs, Fabian; Huttner, Benedikt; Huttner, Angela

    2017-04-12

    Recommendations for the treatment of lower non-catheter-related urinary tract infection (UTI) in men are rarely evidence-based. Their management requires the localization of the site of infection, whether it be the urethra, bladder or prostate, and includes antibiotic therapy and in most cases urological assessment. They are often associated with urinary tract procedures or anatomical or functional abnormalities. Nearly 80 % of male UTIs are caused by Enterobacteriaceae. The prevalence of broad-spectrum beta-lactamase-producing strains (ESBL) and quinolone-resistant strains is increasing. The aim of this article is to define three types of lower, non-catheter-related UTI in men - urethritis, cystitis and prostatitis - their microbiology and management in Switzerland.

  17. Expanding the Repertoire of Carbapenem-Hydrolyzing Metallo-ß-Lactamases by Functional Metagenomic Analysis of Soil Microbiota

    DEFF Research Database (Denmark)

    Gudeta, Dereje D.; Bortolaia, Valeria; Pollini, Simona

    2016-01-01

    as metallo-beta-lactamases (MBLs), were identified in six soil samples, including two subclass B1 (GRD23-1 and SPN79-1) and seven subclass B3 (CRD3-1, PEDO-1, GRD33-1, ESP-2, ALG6-1, ALG11-1, and DHT2-1). Except PEDO-1 and ESP-2, these enzymes were distantly related to any previously described MBLs (33 to 59......% identity). RAIphy analysis indicated that six enzymes (CRD3-1, GRD23-1, DHT2-1, SPN79-1, ALG6-1, and ALG11-1) originated from Proteobacteria, two (PEDO-1 and ESP-2) from Bacteroidetes and one (GRD33-1) from Gemmatimonadetes. All MBLs detected in soil microbiota were functional when expressed in E. coli...

  18. Negative Pressure Wound Therapy in Infected Wound following Posterior Spinal Instrumentation using Simple Self-assembled System: A Case Report

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    CW Chang

    2014-07-01

    Full Text Available Postoperative wound infection in an instrumented spine patient is often disastrous. Management includes implant removal leading to spine instability. Negative pressure wound therapy (NPWT applied to the spine surgical wound is one of the wound care technique with successful results. We report a case of a man who sustained Chance fracture of Lumbar 1 (L1 vertebra treated with long segment posterior instrumentation, who unfortunately developed Extended-spectrum beta-lactamase (ESBL positive E. coli infection one month after the operation. After careful debridement of the wound, the implant became exposed. Three cycles of NPWT were applied and the wound healed with granulation tissue completely covering the implant, and thus negating the need to remove the implant. In conclusion, the NPWT is a good alternative in postoperative wound management especially in an instrumented spine patient.

  19. [New antibacterial agents on the market and in the pipeline].

    Science.gov (United States)

    Kern, W V

    2015-11-01

    After some years of stagnation there have been several new successful developments in the field of antibacterial agents. Most of these new developments have been in conventional antibacterial classes. New drugs among the beta-lactam agents are methicillin-resistant Staphylococcus aureus (MRSA) active cephalosporins (ceftaroline and ceftobiprole) and new combinations of beta-lactam with beta-lactamase inhibitors (ceftolozane/tazobactam, ceftazidime/avibactam, imipenem/relebactam and meropenem/RPX7009). New developments can also be observed among oxazolidinones (tedizolid, radezolid, cadazolid and MRX-I), macrolides/ketolides (modithromycin and solithromycin), aminoglycosides (plazomicin), quinolones (nemonoxacin, delafloxacin and avarofloxacin), tetracyclines (omadacycline and eravacycline) as well as among glycopeptides and lipopeptides (oritavancin, telavancin, dalbavancin and surotomycin). New agents in a very early developmental phase are FabI inhibitors, endolysines, peptidomimetics, lipid A inhibitors, methionyl-tRNA synthetase inhibitors and teixobactin.

  20. Multidrug-resistant opportunistic pathogens challenging veterinary infection control.

    Science.gov (United States)

    Walther, Birgit; Tedin, Karsten; Lübke-Becker, Antina

    2017-02-01

    Although the problems associated with healthcare-associated infections (HAI) and the emergence of zoonotic and multidrug-resistant pathogens in companion animal (dogs, cats and horses) medicine have been well-known for decades, current progress with respect to practical implementation of infection control programs in veterinary clinics has been limited. Clinical outbreak events reported for methicillin-resistant Staphylooccus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP), extended spectrum beta-lactamase (ESBL)-producing Escherichia coli and multidrug-resistant (MDR) Salmonella Serovars indicate the necessity of infection control strategies for protecting animal patients at risk as well as veterinary personnel. The close bond between humans and their companion animals provides opportunities for exchange of microorganisms, including MDR pathogens. This particular aspect of the "One Health" idea requires more representative surveillance efforts and infection control strategies with respect to animal-species specific characters. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. In vitro cost-effective methods to detect carbapenemases in Enterobacteriaceae

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    Varsha Gupta

    2018-01-01

    Full Text Available The rise in carbapenemases-producing organisms has challenged the scientific community. Infections caused by these bacteria have limited treatment options. There are various types such as Klebsiella pneumoniae carbapenemase (Ambler class A, metallo-beta-lactamases of VIM-type, IMP-type, NDM-type (Ambler class B, and OXA-48-types (Ambler class D. An efficient strategy for detection of carbapenemase producers is important to determine the appropriate therapeutic modalities. In this study, four methods - Carba NP test, modified Carba NP (MCNP test, carbapenem inactivation method (CIM test, and Rapidec Carba NP kit test were evaluated. We evaluated an in-house MCNP test to detect carbapenemase production using a single protocol which gave reliable results. Furthermore, CIM using routine antibiotic discs gives good results. Both these tests were found to be cost-effective.

  2. [Antimicrobial therapy in severe infections with multidrug-resistant Gram-negative bacterias].

    Science.gov (United States)

    Duszyńska, Wiesława

    2010-01-01

    Multidrug-resistant Gram-negative bacteria pose a serious and rapidly emerging threat to patients in healthcare settings, and are especially prevalent and problematic in intensive therapy units. Recently, the emergence of pandrug-resistance in Gram-negative bacteria poses additional concerns. This review examines the clinical impact and epidemiology of multidrug-resistant Gram-negative bacteria as a cause of increased morbidity and mortality among ITU patients. Beta-lactamases, cephalosporinases and carbapenemases play the most important role in resistance to antibiotics. Despite the tendency to increased resistance, carbapenems administered by continuous infusion remain the most effective drugs in severe sepsis. Drug concentration monitoring, albeit rarely used in practice, is necessary to ensure an effective therapeutic effect.

  3. Antibacterial Activity of the Extracts Obtained from Rosmarinus officinalis, Origanum majorana, and Trigonella foenum-graecum on Highly Drug-Resistant Gram Negative Bacilli

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    Roula Abdel-Massih

    2010-01-01

    Full Text Available Our aim was to determine the antimicrobial activity of three selected plants (Rosmarinus officinalis, Origanum majorana, and Trigonella foenum-graecum against Extended Spectrum Beta Lactamase (ESBL—producing Escherichia coli and Klebsiella pneumoniae— and to identify the specific plant fraction responsible for the antimicrobial activity. The plants were extracted with ethanol to yield the crude extract which was further subfractionated by different solvents to obtain the petroleum ether, the dichloromethane, the ethyl acetate, and the aqueous fractions. The Minimum Inhibitory Concentrations (MIC and Minimum Bactericidal Concentrations (MBC were determined using broth microdilution. The MICs ranged between 1.25 and 80 g/l. The majority of these microorganisms were inhibited by 80 and 40 g/l of the crude extracts. The petroleum ether fraction of Origanum majorana significantly inhibited 94% of the tested strains. Ethyl acetate extracts of all selected plants exhibited relatively low MICs and could be therefore described as strong antibacterial.

  4. Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104

    DEFF Research Database (Denmark)

    Sandvang, Dorthe; Aarestrup, Frank Møller; Jensen, Lars Bogø

    1997-01-01

    The presence and genetic content of integrons was investigated in eight Salmonella enterica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition...... to the sul1 and qacE Delta 1 genes characteristic of integrons. The first integron encoded the ant (3 ")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-l beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two...... integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements....

  5. Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104

    DEFF Research Database (Denmark)

    Sandvang, Dorthe; Aarestrup, Frank Møller; Jensen, Lars Bogø

    1998-01-01

    The presence and genetic content of integrons was investigated in eight Salmonella enteritica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition...... to the sul1 and qacE Delta 1 genes characteristic of integrons. The first integron encoded the ant (3")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-1 beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two...... integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements....

  6. Development of in vitro transposon assisted signal sequence trapping and its use in screening Bacillus halodurans C125 and Sulfolobus solfataricus P2 gene libraries

    DEFF Research Database (Denmark)

    Becker, F.; Schnorr, K.; Wilting, R.

    2004-01-01

    To identify genes encoding extracytosolic proteins, a minitransposon, TnSig, containing a signal-less beta-lactamase ('bla) as reporter gene, was constructed and used for in vitro transposition of genomic libraries made in Escherichia coli. The 'bla gene was cloned into a bacteriophage MU...... minitransposon enabling translational fusions between 'bla and target genes. Fusion of TnSig in the correct reading frame to a protein carrying transmembrane domains or signal peptides resulted in ampicillin resistance of the corresponding clone. Prokaryotic gene libraries from the alkaliphilic bacterium...... Bacillus halodurans C125 and the hyperthermophilic archaeon Sulfolobus solfataricus P2 were tagged with TnSig. The genomic sequences, which are publicly available (EMBL BA000004 and EMBL AE006641), were used for rapid open reading frame (ORF) identification and prediction of protein localisation...

  7. Comparison of Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Drinking Well Water and Pit Latrine Wastewater in a Rural Area of China

    Directory of Open Access Journals (Sweden)

    Hongna Zhang

    2016-01-01

    Full Text Available The present study was conducted to gain insights into the occurrence and characteristics of extended-spectrum beta-lactamase- (ESBL- producing Escherichia coli (E. coli from drinking well water in the rural area of Laiwu, China, and to explore the role of the nearby pit latrine as a contamination source. ESBL-producing E. coli from wells were compared with isolates from pit latrines in the vicinity. The results showed that ESBL-producing E. coli isolates, with the same antibiotic resistance profiles, ESBL genes, phylogenetic group, plasmid replicon types, and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR fingerprints, were isolated from well water and the nearby pit latrine in the same courtyard. Therefore, ESBL-producing E. coli in the pit latrine may be a likely contributor to the presence of ESBL-producing E. coli in rural well water.

  8. A Case of Healthcare Associated Pneumonia Caused by Chryseobacterium indologenes in an Immunocompetent Patient

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    Salih Atakan Nemli

    2015-01-01

    Full Text Available Chryseobacterium indologenes is nonmotile, oxidase, and indole positive Gram-negative aerobic bacilli which is widely found in plants, soil, foodstuffs, and water. It can colonize hospital environment due to ability to survive in chlorine-treated water supplies. Chryseobacteria can also colonize patients via contaminated medical devices such as respirators, intubation tubes, humidifiers, intravascular catheters, and prosthetic valves. Immune suppression, comorbidities, use of broad-spectrum antibiotics, and extreme age are other important risk factors for Chryseobacterium infections. We report a case of an 82-year-old male admitted to our hospital with the complaint of altered mental status with history of trauma, and recent orthopedic and neurosurgery operations. He was transferred to neurosurgery intensive care unit due to respiratory failure. Urine culture yielded extended spectrum beta lactamase (ESBL (+. E. coli and C. indologenes were isolated from transtracheal aspirate. He was treated with ertapenem, and levofloxacin and discharged with full recovery.

  9. Tawny owl (Strix aluco) as a potential transmitter of Enterobacteriaceae epidemiologically relevant for forest service workers, nature protection service and ornithologists.

    Science.gov (United States)

    Grzywaczewski, Grzegorz; Kowalczyk-Pecka, Danuta; Cios, Szymon; Bojar, Wiktor; Jankuszew, Andrzej; Bojar, Hubert; Kolejko, Marcin

    2017-03-31

    Established taxa within the Enterobacteriaceae wereisolated from cloacal swabs of Strix aluco chicks in nest boxes located at five research sites. ChromID ESBL medium (bioMerieux) was used to select a pool of Enterobacteriaceae strains producing extended-spectrum beta-lactamases. Drug sensitivity of the chosen strains was determined from the full pool of Enterobacteriaceae to 6 chemotherapeutics of different mechanisms of action. The study evaluated the sensitivity of ESBL-synthesizing isolates to substances belonging to penicillins, cephalosporins, cephamycins, clavams, carbapenems and monobactams. Analysis of the results indicated a potential role of Strix aluco in the dissemination of epidemiologically-relevant Enterobacteriaceae, and, importantly, pose health risks to forest service workers, nature protection service and ornithologists. The results can also serve as the basis for further environmental studies.

  10. Bacterial Cheating Limits the Evolution of Antibiotic Resistance

    Science.gov (United States)

    Yurtsev, Eugene; Xiao Chao, Hui; Datta, Manoshi; Artemova, Tatiana; Gore, Jeff

    2012-02-01

    The emergence of antibiotic resistance in bacteria is a significant health concern. Bacteria can gain resistance to the antibiotic ampicillin by acquiring a plasmid carrying the gene beta-lactamase, which inactivates the antibiotic. This inactivation may represent a cooperative behavior, as the entire bacterial population benefits from removal of the antibiotic. The presence of a cooperative mechanism of resistance suggests that a cheater strain - which does not contribute to breaking down the antibiotic - may be able to take advantage of resistant cells. We find experimentally that a ``sensitive'' bacterial strain lacking the plasmid conferring resistance can invade a population of resistant bacteria, even in antibiotic concentrations that should kill the sensitive strain. We use a simple model in conjunction with difference equations to explain the observed population dynamics as a function of cell density and antibiotic concentration. Our experimental difference equations resemble the logistic map, raising the possibility of oscillations or even chaotic dynamics.

  11. Bacterial cheating limits antibiotic resistance

    Science.gov (United States)

    Xiao Chao, Hui; Yurtsev, Eugene; Datta, Manoshi; Artemova, Tanya; Gore, Jeff

    2012-02-01

    The widespread use of antibiotics has led to the evolution of resistance in bacteria. Bacteria can gain resistance to the antibiotic ampicillin by acquiring a plasmid carrying the gene beta-lactamase, which inactivates the antibiotic. This inactivation may represent a cooperative behavior, as the entire bacterial population benefits from removing the antibiotic. The cooperative nature of this growth suggests that a cheater strain---which does not contribute to breaking down the antibiotic---may be able to take advantage of cells cooperatively inactivating the antibiotic. Here we find experimentally that a ``sensitive'' bacterial strain lacking the plasmid conferring resistance can invade a population of resistant bacteria, even in antibiotic concentrations that should kill the sensitive strain. We observe stable coexistence between the two strains and find that a simple model successfully explains the behavior as a function of antibiotic concentration and cell density. We anticipate that our results will provide insight into the evolutionary origin of phenotypic diversity and cooperative behaviors.

  12. Quantum-Sequencing: Fast electronic single DNA molecule sequencing

    Science.gov (United States)

    Casamada Ribot, Josep; Chatterjee, Anushree; Nagpal, Prashant

    2014-03-01

    A major goal of third-generation sequencing technologies is to develop a fast, reliable, enzyme-free, high-throughput and cost-effective, single-molecule sequencing method. Here, we present the first demonstration of unique ``electronic fingerprint'' of all nucleotides (A, G, T, C), with single-molecule DNA sequencing, using Quantum-tunneling Sequencing (Q-Seq) at room temperature. We show that the electronic state of the nucleobases shift depending on the pH, with most distinct states identified at acidic pH. We also demonstrate identification of single nucleotide modifications (methylation here). Using these unique electronic fingerprints (or tunneling data), we report a partial sequence of beta lactamase (bla) gene, which encodes resistance to beta-lactam antibiotics, with over 95% success rate. These results highlight the potential of Q-Seq as a robust technique for next-generation sequencing.

  13. Staphylococcus aureus ampicillin-resistant from the odontological clinic environment Staphylococcus aureus resistente à ampicilina em ambiente de Clínica Odontológica

    Directory of Open Access Journals (Sweden)

    Wagner Luis de Carvalho Bernardo

    2005-02-01

    Full Text Available The aim of this research was to evaluate the prevalence of Sthaphylococcus spp. and S. aureus in the odontological clinic environment (air, their production of beta-lactamase and antibacterial susceptibility to the major antibiotics utilized in medical particle. During 12 months of samples collect were isolated 9775 CFU by MSA medium suggesting a high amount of Staphylococcus spp. in the clinic environment which can appear through aerosols. A total of 3149 colonies (32.2% were suggestive of pathogenic staphylococci. Gram coloration, catalase test, colony-mallow growing on chromogenic medium, and coagulase test confirmed the identity of 44 (0.45% S. aureus isolates. Of these, 35 isolates (79.5% showed production of beta-lactamase by CefinaseTM discs and resistance to ampicillin, erythromycin (7 isolates and tetracycline (1 isolate suggesting the existence of multiresistant isolates. The evaluation of the oxacillin MIC by Etest® assays showed susceptibility patterns suggesting the inexistence of the mecA gene in chromosomal DNA. These results point out to the need of a larger knowledge on the contamination means and propagation of this microorganism into the odontological clinic.Foi avaliada a prevalência de Staphylococcus spp. e S. aureus no ambiente clínico odontológico, a produção de beta-lactamase e a susceptibilidade antibacteriana aos principais antibióticos utilizados na prática clínica. Durante 12 meses de coleta de amostras foram isolados 9775 UFC no meio de cultura AMS, demonstrando uma elevada quantidade de Staphylococcus spp. no ambiente clínico, provavelmente em decorrência da propagação de aerossóis. Um total de 3149 colônias (32,2% foi sugestivo de estafilococos patogênicos. Coloração de Gram, teste de catalase, crescimento de colônias-malva sobre meio cromogênico e teste de coagulase confirmaram a identidade de 44 (0,45% isolados de S. aureus. Destes, 35 isolados (79,5% mostraram produção de beta-lactamase

  14. [Arms racing between human beings and pathogens: NDM-1 and superbugs].

    Science.gov (United States)

    Sun, Mingwei; Zheng, Beiwen; Gao, George F; Zhu, Baoli

    2010-11-01

    Throughout human history, pandemic bacterial diseases such as the plague and tuberculosis have posed an enormous threat to human beings. The discovery of antibiotics has provided us with powerful arsenal for the defense against bacterial infections. However, bacteria are acquiring more and more resistance genes to shield off antibiotics through mutation and horizontal gene transfer. Therefore, novel antibiotics must be produced and the arms race between bacterial pathogens and antibiotics is becoming increasingly intense. Recently, researchers have found that plasmids carrying a new metallo-beta-lactamase gene, blaNDM-1, and many other antibiotics resistance genes can easily spread through bacterial populations and confer recipient stains resistance to nearly all of the current antibiotics. It is a threat to the human health and a great challenge for our medical science, which we are facing. We need to find new ways to fight and win this arms racing.

  15. Whole genome sequence to decipher the resistome of Shewanella algae, a multidrug-resistant bacterium responsible for pneumonia, Marseille, France.

    Science.gov (United States)

    Cimmino, Teresa; Olaitan, Abiola Olumuyiwa; Rolain, Jean-Marc

    2016-01-01

    We characterize and decipher the resistome and the virulence factors of Shewanella algae MARS 14, a multidrug-resistant clinical strain using the whole genome sequencing (WGS) strategy. The bacteria were isolated from the bronchoalveolar lavage of a hospitalized patient in the Timone Hospital in Marseille, France who developed pneumonia after plunging into the Mediterranean Sea. The genome size of S. algae MARS 14 was 5,005,710 bp with 52.8% guanine cytosine content. The resistome includes members of class C and D beta-lactamases and numerous multidrug-efflux pumps. We also found the presence of several hemolysins genes, a complete flagellum system gene cluster and genes responsible for biofilm formation. Moreover, we reported for the first time in a clinical strain of Shewanella spp. the presence of a bacteriocin (marinocin). The WGS analysis of this pathogen provides insight into its virulence factors and resistance to antibiotics.

  16. Radioimmunoassay and enzyme-linked immunoassay of antibodies directed against lymphadenopathy-associated virus (LAV) proteins larger than the core protein (P24)

    International Nuclear Information System (INIS)

    Neurath, A.R.; Strick, N.; Lee, Y.S.; Nilsen, T.; Baker, L.; Sproul, P.; Rubinstein, P.; Taylor, P.; Stevens, C.E.; Gold, J.W.M.

    1985-01-01

    Molecular exclusion chromatography of crude LAV antigen preparations allows separation of most of P24 from larger proteins of LAV (PL). PL and 125 I- or beta-lactamase-labeled anti-LAV were used as reagents for radioimmunoassay (RIA) - or enzyme-linked immunoassay (ELISA) - inhibition tests to detect antibodies directed predominantly against PL (anti-PL). Among 257 individuals belonging to groups at high risk of developing AIDS, 117 (45.5%) were positive for anti-PL and 108 (42%) for anti-P24, respectively. The 2 individuals among 600 random blood donors found to be anti-P24-positive in the preceding study also had anti-PL in their serum. Sera from 500 additional blood donors were screened for anti-PL and 1 of these was positive. The implication of these findings for screening of blood donors is discussed. (Auth.)

  17. Radioimmunoassay and enzyme-linked immunoassay of antibodies directed against lymphadenopathy-associated virus (LAV) proteins larger than the core protein (P24)

    Energy Technology Data Exchange (ETDEWEB)

    Neurath, A R; Strick, N; Lee, Y S; Nilsen, T; Baker, L; Sproul, P; Rubinstein, P; Taylor, P; Stevens, C E; Gold, J W.M.

    1985-10-01

    Molecular exclusion chromatography of crude LAV antigen preparations allows separation of most of P24 from larger proteins of LAV (PL). PL and /sup 125/I- or beta-lactamase-labeled anti-LAV were used as reagents for radioimmunoassay (RIA) - or enzyme-linked immunoassay (ELISA) - inhibition tests to detect antibodies directed predominantly against PL (anti-PL). Among 257 individuals belonging to groups at high risk of developing AIDS, 117 (45.5%) were positive for anti-PL and 108 (42%) for anti-P24, respectively. The 2 individuals among 600 random blood donors found to be anti-P24-positive in the preceding study also had anti-PL in their serum. Sera from 500 additional blood donors were screened for anti-PL and 1 of these was positive. The implication of these findings for screening of blood donors is discussed. 17 refs.; 2 figs.; 1 table.

  18. Bacterial resistance to antimicrobial agents in Latin America. The giant is awakening.

    Science.gov (United States)

    Guzmán-Blanco, M; Casellas, J M; Sader, H S

    2000-03-01

    Resistant bacteria are emerging in Latin America as a real threat to the favorable outcome of infections in community- and hospital-acquired infections. Despite present extensive surveillance, healthcare workers who most need the information may be unaware of this growing problem. Outbreaks of meningococci with diminished susceptibility to penicillin have been reported in the region; a constant increase of resistance to penicillin in pneumococci and poor activity of commonly used oral antibiotics for the treatment of community-acquired urinary tract infections have made the treatment of these infections more difficult. Reports from tertiary hospitals are similar to many other areas of the world, with increasing frequency of Klebsiella pneumoniae-carrying extended-spectrum beta-lactamase, multiresistant strains of Pseudomonas aeruginosa and Acinetobacter baumanni in ICU settings, and reports of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. A surveillance network readily accessible to those who prescribe antibiotics in Latin America is highly desirable.

  19. PER-1 production in a urinary isolate of Providencia rettgeri.

    Science.gov (United States)

    Bahar, G; Eraç, B; Mert, A; Gülay, Z

    2004-08-01

    A Providencia rettgeri strain resistant to extended-spectrum cephalosporins and intermediate to aztreonam was isolated from the urine of a patient hospitalized in the urology clinic of SSK Educational Hospital in Ankara. Clavulanic acid restored the activity of extended-spectrum cephalosporins, suggesting that the strain was harboring an extended-spectrum beta-lactamase. Since the PER-1 enzyme is widespread in Turkey, and had been already detected in a related species such as Proteus mirabilis, the Providencia strain was suspected of harboring a PER-1 enzyme, which was indeed detected by PCR. This is the first description in a P. rettgeri isolate of a PER-1 enzyme which is widespread among Acinetobacter baumanni and Pseudomonas aeruginosa strains in Turkey.

  20. Doripenem: position in clinical practice.

    Science.gov (United States)

    Dedhia, Harakh V; McKnight, Richard

    2009-06-01

    Doripenem is a novel carbapenem with a broad spectrum of activity against Gram-positive pathogens, anerobes and Gram-negative bacteria, including Pseudomonas aeruginosa. Doripenem exhibits rapid bactericidal activity with two- to fourfold lower MIC values for Gram-negative bacteria, compared with other carbapenems such as imipenem. Doripenem is approved for the treatment of complicated intra-abdominal infection and urinary tract infections. It has been successfully used in the treatment of nosocomial and ventilator-associated pneumonia. It has a potential to be the drug of choice for these conditions. This evaluation focuses on the general review of the drug, including mechanisms of resistance, clinical efficacy and the position of doripenem in clinical practice. Stability against numerous beta-lactamases, low adverse-event potential and more potent in vitro and possibly in vivo activity against P. aeruginosa and Acinetobacter baumanni compared with existing carbapenems are attractive features.

  1. Antibiotic Resistance in Pediatric Urinary Tract Infections.

    Science.gov (United States)

    Stultz, Jeremy S; Doern, Christopher D; Godbout, Emily

    2016-12-01

    Urinary tract infections (UTIs) are a common problem in pediatric patients. Resistance to common antibiotic agents appears to be increasing over time, although resistance rates may vary based on geographic region or country. Prior antibiotic exposure is a pertinent risk factor for acquiring resistant organisms during a first UTI and recurrent UTI. Judicious prescribing of antibiotics for common pediatric conditions is needed to prevent additional resistance from occurring. Complex pediatric patients with histories of hospitalizations, prior antibiotic exposure, and recurrent UTIs are also at high risk for acquiring UTIs due to extended spectrum beta-lactamase-producing organisms. Data regarding the impact of in vitro antibiotic susceptibility testing interpretation on UTI treatment outcomes is lacking.

  2. CLINICAL CHARACTERISTICS AND ANTIBIOTIC RESISTANCE PATTERN OF PATHOGENS IN PEDIATRIC URINARY TRACT INFECTION.

    Science.gov (United States)

    Amornchaicharoensuk, Yupaporn

    2016-09-01

    Medical records of children less than 15-years of age admitted to hospital for urinary tract infection (UTI) from January 2010 to December 2014 were reviewed. Among 100 children (59% males and 41% females) with upper UTI, the most common pathogen (88%) was Escherichia coli, of which 69% were nonextended spectrum beta-lactamase (ESBL) and 19 % ESBL producers. Resistance to ampicillin and trimethoprim/sulfamethoxazole was 90% and 60%, respectively. All ESBL-producing E. coli were resistant to ampicillin and third generation cephalosporins (cefotaxime and ceftriaxone), while 87% and 1.5% of non ESBL-producing E. coli were resistant to ampicillin and the two third generation cephalosporins, respectively. These data highlight the high prevalence of ESBL-producing E. coli in pediatric UTI and the potential problem in treating such infections.

  3. Resistência bacteriana e ação das bacteriocinas de Lactobacillus spp em Staphylococcus aureus isolados de mastite bovina

    Directory of Open Access Journals (Sweden)

    B.R. Pribul

    2011-06-01

    Full Text Available Staphylococcus aureus is the major pathogen causing intramammary infections in dairy cattle worldwide. Among the factors that contribute to its spread and infectious potential is the ability to overcome the mechanisms of antimicrobials activity. The present work investigated the antimicrobial resistance pattern and sensibility to bacteriocins produced by strains of Lactobacillus spp of 30 isolates of S. aureus from mastitis. From this, 29 are beta-lactamase producers. Eight isolates (26.6% showed resistance to at least four antibiotics being considered multiresistent. All of them were mecA-positive. Otherwise, all isolates tested showed sensibility to at least one of the four bacteriocin producer strains. Due to the significant depletion of the efficacy of antimicrobials, pathogen growth inhibition by bacteriocins seems an alternative of biological control in infectious processes.

  4. Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP

    DEFF Research Database (Denmark)

    Taponen, S.; Simojoki, H.; Haveri, M.

    2006-01-01

    The coagulase-negative staphylococcal species causing mastitis in lactating cattle were identified and possible differences in the clinical characteristics or persistence of mastitis caused by different CNS were evaluated. The effect of antimicrobial treatment was also assessed. In addition, AFLP...... of these species. Approximately half of the mastitis cases were clinical, and in the majority clinical signs were mild. The severity and persistence of intramammary infection were unaffected by CNS species. Fifty-nine percent of the quarter cases were treated with antimicrobials, and the rest were left without...... treatment. Mastitis due to P-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were P-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45...

  5. Case report: liver abscess pyogenic after peritonitis appendix perforation

    Science.gov (United States)

    Damanik, E. H.; Ginting, F.

    2018-03-01

    Two of the most common liver abscess is anamoebic liver abscess and pyogenic liver abscess (PLA). PLA could be as singular or multiple abscesses. It is usually caused by Klebsiella pneumonia and Escherichia coli. Historically, PLA is usually caused by acute appendicitis, but with developed of surgical practice and microbiology, the number of events has decreased. Here we present a case of a39-year-old woman that developed a PLA after she had an appendectomy about six months ago. An ultrasonogram and abdominal scan showed an abscess in the right lobe. We performed paracentesis, and the result from the pus culturewas positive for Escherichia coli with Extended-Spectrum Beta-Lactamase (ESBL) (+) that showed the same as the culture from lesion taken from her appendix. This report emphasizes the fact that, nowadays we still found Pyogenic liver abscess after peritonitis appendix perforation.

  6. Nosocomial bloodstream infections in a Turkish university hospital: study of Gram-negative bacilli and their sensitivity patterns.

    Science.gov (United States)

    Köseoğlu , O; Kocagöz, S; Gür, D; Akova, M

    2001-06-01

    Treatment of nosocomial bacteraemia is usually governed by the surveillance results of the particular unit. Such results are especially important when antimicrobial resistance rates are high. Multiresistant isolates including Gram-negatives producing extended-spectrum beta-lactamases have been frequently reported in tertiary care units in Turkey. In this study, antimicrobial susceptibilities of Gram-negative blood isolates (n=348) were determined by microbroth dilution tests. The results showed carbapenems (meropenem and imipenem) to be uniformly more potent in vitro than any other drug against the Enterobacteriaceae. Quinolone antibiotics were more active in vitro than aminoglycosides against a range of bacteria. Gram-negative bloodstream isolates were highly resistant to many antimicrobial agents in the hospital. In order to prevent hospital infection and antimicrobial resistance, surveillance of aetiological agents must be performed regularly.

  7. Reversal of methicillin resistance in Staphylococcus aureus by thioridazine

    DEFF Research Database (Denmark)

    Klitgaard, Janne K; Skov, Marianne N; Kallipolitis, Birgitte H

    2008-01-01

    of thioridazine in the presence of a fixed amount of oxacillin. Furthermore, the protein level of PBP2a was reduced when bacteria were treated with the combination of oxacillin and thioridazine. The two drugs also affected the mRNA level of the beta-lactamase gene, blaZ. Conclusions The present study indicates......Objectives Thioridazine has been shown to reverse oxacillin resistance in methicillin-resistant Staphylococcus aureus (MRSA) in vitro. The aim of this study was to investigate whether thioridazine alone or in combination with oxacillin affects the transcription of the methicillin resistance gene...... blotting in the presence of thioridazine and oxacillin. Results We observed an increased susceptibility of MRSA towards oxacillin in the presence of thioridazine compared with bacteria grown with oxacillin or thioridazine alone. Transcription of mecA was reduced with increasing concentrations...

  8. Antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and group A beta-haemolytic streptococci in 2002-2003. Results of the multinational GRASP Surveillance Program

    DEFF Research Database (Denmark)

    Beekmann, Susan E; Heilmann, Kris P; Richter, Sandra S

    2005-01-01

    A multinational surveillance study, GRASP, was conducted between November 2002 and April 2003 with the aim of assessing rates of antimicrobial resistance among 2656 isolates of Streptococcus pneumoniae, 2486 isolates of group A beta-haemolytic streptococci, 1358 isolates of Haemophilus influenzae...... and 1047 of Moraxella catarrhalis from 20 countries in Europe, eastern Asia and southern Africa. Conspicuous differences between various countries were noted in the S. pneumoniae resistance rates observed for penicillin (0-79.2%) and erythromycin (4-66%), along with other antimicrobials. The percentage...... of MDR strains was above 25% in 8 of the 20 countries studied. Group A streptococcal macrolide resistance rates ranged from 0% to 35% by country, while rates of beta-lactamase production ranged from 0% to 39% for H. influenzae and 80-100% for M. catarrhalis. Antibiotic resistance in S. pneumoniae remains...

  9. Human Intestinal Cells Modulate Conjugational Transfer of Multidrug Resistance Plasmids between Clinical Escherichia coli Isolates

    DEFF Research Database (Denmark)

    Machado, Ana Manuel; Sommer, Morten

    2014-01-01

    Bacterial conjugation in the human gut microbiota is believed to play a major role in the dissemination of antibiotic resistance genes and virulence plasmids. However, the modulation of bacterial conjugation by the human host remains poorly understood and there is a need for controlled systems...... to study this process. We established an in vitro co-culture system to study the interaction between human intestinal cells and bacteria. We show that the conjugation efficiency of a plasmid encoding an extended spectrum beta-lactamase is reduced when clinical isolates of Escherichia coli are co...... of the intestinal cells exposed to bacteria leading to a two-fold reduction in conjugation efficiency. These results show that human gut epithelial cells can modulate bacterial conjugation and may have relevance to gene exchange in the gut....

  10. STUDY ON SURGICAL SITE INFECTIONS CAUSED BY ESBL PRODUCING GRAM NEGATIVE BACTERIA

    Directory of Open Access Journals (Sweden)

    Rambabu

    2015-09-01

    Full Text Available Surgical site infections have been a major problem, because of the emergence of drug resistant bacteria, in particular B - lactamase producing bacteria. Extended spectrum beta lactamase producing gram negative organisms pose a great challenge in treatment o f SSI present study is aimed at determining multiple drug resistance in gram negative bacteria & to find out ESBL producers, in correlation with treatment outcome. A total of 120 wound infected cases were studied. Staphylococcus aureus was predominant bact erium - 20.Among gram negative bacteria, Pseudomonas species is predominant (14 followed by Escherichia coli (13 , Klebsiella species (12 , Proteus (9 Citrobacter (4 Providencia (2 & Acinetobacter species (2 . Out of 56 gramnegative bacteria isolated, 20 were i dentified as ESBL producers, which was statistically significant. Delay in wound healing correlated with infection by ESBL producers, which alarms the need of abstinence from antibiotic abuse

  11. Aspects of constitutive and acquired antibioresistance in Aeromonas hydrophila strains isolated from water sources.

    Science.gov (United States)

    Balotescu, Carmen; Israil, Anca; Radu, Roxana; Alexandru, Ionela; Dobre, Georgeta

    2003-01-01

    Over the last three decades, the literature pointed out the implications of Aeromonas species in human pathology. These species were described as being involved in intestinal (several outbreaks of acute gastroenteritis of choleric/dysenteric form or chronic diarrhoea, ulcerative colitis, etc.) in normal adults or children, as well as in extraintestinal infections in immunocompromised hosts. This last aspect included a large range of cutaneous injuries (micronecrosis, abscesses, bums, cellulites, furunculosis), joint, bones, respiratory, urinary tract, ocular infections up to meningitis, endocarditis, peritonitis, hepatobilliary disease, endotoxic shock and septicemia (as consequence of leech microvascular surgery). During the last decade, the literature reported a high mortality in Aeromonas infections determined by certain phenospecies (A. hydrophila and A. veronii) especially in extraintestinal infections in immunocompromised patients. In microbiologists' opinion this high rate of mortality was probably due to poor knowledge concerning the aspects of antibioresistance in Aeromonas strains, to empiric treatments with antibiotics to which these bacteria exhibiting constitutive resistance lead to insuccessful results, and at last to the increasing trend of aeromonads resistance to certain antibiotics after 1996. The literature mentioned also that for a great number of Beta-lactamase producing Aeromonas strains, the use of microdilution method (by comparison to disk diffusion in agar medium) giving false results made more difficult the true knowledge of Aeromonas antibioresistance patterns. At the same time, in 2002, the literature mentioned 4 ecological compartments considered as "reservoirs for dissemination and transfer of microbial antibioresistance i.e. humans, animals, plants and natural soil and water. In the last time, more and more data of the literature revealed that some bacteria with role of reservoir of antibioresistance in the natural environment, even

  12. Bile anaerobic bacteria detection and antibiotic susceptibility in patients with gallstone.

    Science.gov (United States)

    Lu, Yun; Xiang, Ting-Hai; Shi, Jing-Sen; Zhang, Bing-Yuan

    2003-08-01

    To detect bile anaerobic bacteria and antibiotic susceptibility in 59 patients with gallstones who had had cholecystectomy. BACT/ALERT 120 microbe detection system and SCEPTOR microbe detection system were used to detect bile anaerobic bacteria, antibiotic susceptibility. The ratio of anaerobic bacteria to the patients examined was 52.5% (31/59). Obligate anaerobe bile culture showed positive results in 4 patients. B. fragilis (37.8%) was the major type of anaerobic bacteria in bile. Most (81.8%) of anaerobic bacteria were sensitive to metronidazole, and imipenem was suitable for beta-lactamase bacteria. Culture of anaerobic bacteria in logarithmic phase can improve the positive rate of the culture. There are some relations between anaerobic infection and gallstone formation.

  13. Update on the taxonomy and the clinical and laboratory characteristics of pigmented anaerobic gram-negative rods.

    Science.gov (United States)

    Jousimies-Somer, H R

    1995-06-01

    Pigmented anaerobic gram-negative rods are currently categorized as 17 species distributed in three genera: Prevotella, Porphyromonas, and Bacteroides. These organisms are often encountered in clinical specimens but are also found as part of the indigenous flora on various mucosal surfaces. Several studies are presently assessing the association of individual species with health and disease. For example, Porphyromonas gingivalis and Porphyromonas endodontalis are key putative pathogens in adult periodontitis and root canal infections, respectively. Porphyromonas asaccharolytica is prevalent in extraoral infections. The Porphyromonas species of animal origin have been isolated from infected bite wounds in humans. Isolates closely resembling Bacteroides levii have been recovered from various types of human infections. According to preliminary reports, Prevotella intermedia tends to be associated more often with periodontal disease than with a healthy oral cavity. In the laboratory, enzyme profiling facilitates the identification of these pigmented rods. Beta-Lactamase production is more common among prevotella species (30%-50%) than among Porphyromonas species (< 10%).

  14. ESBL-Producing Escherichia coli

    DEFF Research Database (Denmark)

    Hertz, Frederik Boetius

    Urinary tract infection (UTI) is one the most common bacterial infections and is regularly treated in primary health care. The most common cause of UTI is extraintestinal pathogenic Escherichia coli (ExPEC) already present in the intestinal microflora, often as the dominating strain. Resistance...... in E.coli is increasing and especially isolates producing Extended-Spectrum Beta-Lactamases (ESBL) have been reported worldwide. Treatment of UTI is usually initiated by the general practitioners and a significant proportion of clinical isolates are now resistant to first line antibiotics. The global...... to investigate (i) antibiotics involved in selection of ESBL-producing E.coli, in an experimental mouse model in vivo, (ii) risk factors for UTI with ESBL-producing E.coli and (iii) to describe the phylogenetic composition of E.coli populations with different resistance patterns. We found that different...

  15. Efficacy and safety of Sultamicillin (Ampicillin/Sulbactan) and Amoxicillin/Clavulanic acid in the treatment of upper respiratory tract infections in adults--an open-label, multicentric, randomized trial.

    Science.gov (United States)

    Ferreira, João Batista; Rapoport, Priscila Bogar; Sakano, Eulália; Kós, Arthur Octávio De Avila; Piltcher, Otávio B; Pignatari, Shirley Shizue Nagata; Pinheiro, Sebastião Diógenes; Mocellin, Marcos

    2006-01-01

    Upper respiratory tract infections are the most common causes of medical visits in children and adults, demanding massive use of antibiotics. Bacterial resistance caused by beta-lactamase is one of the most serious problems in this matter. Sultamicillin, a double pro-drug of Ampicillin/Sulbactan, is a potent beta-lactamase inhibitor which can face this challenge. Evaluate efficacy, safety and tolerability of Ampicillin/Sulbactan compared to Amoxicillin/Clavulanate in upper respiratory tract infections in adults. 102 patients were enrolled and randomized to receive Ampicillin/Sulbactan or Amoxicillin/Clavulanate during 10 days. They were evaluated 10 and 30 days after treatment to learn about the therapeutic response. There were no differences between the two groups respecting cure at the end of treatment (visit 2) or at the end of the study (visit 3). Cure ratio was 61.7% and 93.2% (visits 2 and 3) in the Amoxicillin/Clavulanate group compared to 64.4% and 97.4%, respectively, in Ampicillin/Sulbactan group. The adverse events ratio for the two groups was the same (p=0.940). The number of patients with diarrhea was greater in the group of patients receiving Amoxicillin/Clavulanate (70.6%) than in the group receiving Ampicillin/Sulbactan (29.4%) (p=0.0164). Ampicillin/Sulbactan is as safe and efficient as Amoxicillin/Clavulanate in the empiric treatment of upper respiratory infections in adults. The low occurrence of diarrhea in the group receiving Ampicillin/Sulbactan needs confirmation in other studies.

  16. In vitro and in vivo activities of SCE-2787, a new parenteral cephalosporin with a broad antibacterial spectrum.

    Science.gov (United States)

    Iwahi, T; Okonogi, K; Yamazaki, T; Shiki, S; Kondo, M; Miyake, A; Imada, A

    1992-01-01

    SCE-2787, a new cephalosporin having a condensed azolium moiety in the 3 position and an aminothiadiazolyl group in the 7 beta side chain, was evaluated for its in vitro and in vivo activities in comparison with those of ceftazidime, flomoxef, cefpirome, and E1040. Against methicillin-susceptible strains of Staphylococcus aureus and Staphylococcus epidermidis, SCE-2787 was more active than ceftazidime and E1040 and was as active as flomoxef and cefpirome, with MICs for 90% of strains tested (MIC90s) being 1.56 micrograms/ml or less. SCE-2787 was also active against Pseudomonas aeruginosa, for which the MIC90 was 6.25 micrograms/ml, which was lower than that of cefpirome and comparable to that of ceftazidime. SCE-2787 was marginally active against methicillin-resistant strains of staphylococci and Enterococcus faecalis, although its MIC90s were the lowest among those of the antibiotics tested. The activities of SCE-2787 against Streptococcus species, most members of the family Enterobacteriaceae, and Haemophilus influenzae exceeded those of ceftazidime and flomoxef and were comparable to those of cefpirome. Furthermore, MIC90s of SCE-2787 were significantly lower than those of ceftazidime for ceftazidime-resistant isolates of Citrobacter freundii and Enterobacter cloacae. SCE-2787 was resistant to hydrolysis by various types of beta-lactamases, including the Bush group 1 beta-lactamases, and had low affinities for these enzymes, with Km or Ki values of greater than 100 microM. The in vitro activity of SCE-2787 was reflected in its efficacy in mouse protection tests. Thus, SCE-2787 appears to be a promising cephalosporin that should be further evaluated in clinical trials. PMID:1510428

  17. Detection of Ambler class A, B and D ß-lactamases among Pseudomonas aeruginosa and Acinetobacter baumannii clinical isolates from burn patients.

    Science.gov (United States)

    Hakemi Vala, M; Hallajzadeh, M; Hashemi, A; Goudarzi, H; Tarhani, M; Sattarzadeh Tabrizi, M; Bazmi, F

    2014-03-31

    In this study, we evaluated the existence of classes A, B and D ß-lactamases among Pseudomonas aeruginosa (P.aeruginosa) and Acinetobacter baumannii (A.baumannii) strains isolated from burn patients in Tehran during the years 2012 and 2013. From these strains, the frequency of MBL (metallo-beta-lactamase) and ESBL (extended-spectrum beta-lactamase) producers were evaluated using CDDT (Combined Disk Diffusion Tests). The prevalence of some related genes, including blaIMP, blaVIM, blaSPM, blaKPC, blaGIM, blaDIM, blaBIC, blaOXA-48, blaCTX-M-15 and blaNDM genes, was evaluated using PCR and sequencing methods. Of the 75 non-fermenter isolates, 47 P.aeruginosa and 28 A.baumannii were isolated and identified. A high rate of resistance to common antibiotics was detected among A.baumannii isolates in particular, showing 100% resistance to 9 tested antibiotics. CDDT showed that 21 (28%) and 25 (34.25%) of the non-fermenter isolates were ESBL and MBL producers respectively. The prevalence of blaCTX-M-15 and blaIMP genes among the 75 non-fermenter isolates was 7 (9.3%) and 1 (1.3%), respectively. Fortunately, no other genes were detected in either of the non-fermenters. The mortality rate due to MBL-producing isolates was 5 (20%). This study showed specific resistance genes exist among some MBL and ESBL gram-negative non-fermenters which were isolated from burn patients in Tehran.

  18. Genetic Characterization of Carbapenem-Resistant Enterobacteriaceae and the Spread of Carbapenem-Resistant Klebsiella pneumonia ST340 at a University Hospital in Thailand

    Science.gov (United States)

    Netikul, Thidarat; Kiratisin, Pattarachai

    2015-01-01

    Carbapenem-resistant Enterobacteriaceae (CRE) has increasingly spread worldwide in the past decade. The prevalence and characteristics of CRE in Thailand are unknown. In this study, we conducted a 2-year surveillance of CRE among 12,741 clinical isolates of Enterobacteriaceae at the largest university hospital in Thailand with molecular characterization of beta-lactamase (bla) genes, including carbapenemase genes. The CRE prevalence was 1.4%. blaKPC-13 and blaIMP-14a were the only carbapenemase genes detected among these CRE isolates. blaKPC-13 gene was found in a single isolate of Escherichia coli, Enterobacter cloacae and Citrobacter freundii, and blaIMP-14a was found in four isolates of Klebsiella pneumoniae. Carbapenem-resistant K. pneumoniae (CRKP) isolates were resistant to multiple carbapenems at a higher ratio than other CRE species, and thus were further characterized for resistance phenotypes, bla genotypes and molecular epidemiology. Most CRKP isolates harboured multiple bla genes, especially those related to extended-spectrum beta-lactamases. Seven CRKP isolates were resistant to all tested carbapenems, and showed decreased ompK35 and/or ompK36 porin gene expression. Molecular typing of CRKP based on pulsed-field gel electrophoresis (PFGE) demonstrated several unrelated clones. Multilocus sequence typing (MLST) was partially concordant with PFGE results and revealed that ST340, a member of drug-resistant K. pneumoniae clonal complex 258, was the most predominant clone, followed by ST48, ST11 and ST273. The novel ST1645 was identified from this study. ST340 has neither been shown to be predominated among CRKP from other studies, nor been reported in Thailand. Therefore, it emphases a critical concern to monitor and control the spread of CRKP. PMID:26407326

  19. Study of Prevalence and Characteristics of Staphylococcus aureus and CA-MRSA Nasal Colonization in 2-5 Years Old Children in Isfahan

    Directory of Open Access Journals (Sweden)

    Ramin Dibaj

    2014-10-01

    Full Text Available Background and Aim: We carried out a descriptive study to determine the extent of nasal colonization and characteristics of Staphylococcus aureus and CA-MRSA isolates in 2-5 year old children of day care centers in Isfahan. Materials and Methods: The characteristics of isolates were determined using standard phenotypic profiles including colony morphology, Gram staining, catalase, hyaluronidase, coagulase and Dnase tests as well as mannitol fermentation. The MRSA detection was carried out according to CLSI guidelines with oxacillin agar screen test. Methicillin resistance was further confirmed by detection of a 310 bp fragment of mecA gene of MRSA by PCR. Drug susceptibility testing to antibiotics other than methicillin was conducted by disk diffusion. The Beta-lactamase production and inducible clindamycin resistance were also determined by performing the double-disc diffusion(D-test. Results: Out of 323 children, 115 (35.6% carried S. aureus and 11 (9.5% carried MRSA. All MRSA strains were found to contain mecA gene. The susceptibility of strains to vancomycin, rifampicin and Linezolid were 100%. The susceptibility of strains to gentamicin, clindamycin, erythromycin, co-trimoxazole, amoxiclav, ciprofloxacin, tetracycline and penicillin were 99%, 97%, 94%, 94%, 93%, 88%, 44.4% and 1.8% respectively. Beta-lactamase production was seen in 19 (16.5% of staphylococcal strains. Inducible clindamycin resistance was seen in 4 (3.5% of the isolates. Conclusions: Our data indicates that the spread of CA-MRSA within Iranian population is worthy of consideration and merits further molecular investigation to determine the source and mode of transmission.

  20. Recent advances in the chemistry and biology of carbapenem antibiotics.

    Science.gov (United States)

    Coulton, S; Hunt, E

    1996-01-01

    The discovery of the olivanic acids and thienamycin aroused considerable interest amongst medicinal chemists and microbiologists around the world. The susceptibility of these agents to metabolic degradation has, however, been a major obstacle in their development. For many years the only notable success from such intensive research was the combination of imipenem with cilastatin, an inhibitor of the renal dipeptidase enzyme DHP-1. The enormous success of Primaxin for the treatment of a range of life-threatening bacterial infections provided the impetus for the discovery of totally synthetic, non-natural carbapenem derivatives that combine the broad spectrum of antimicrobial activity with stability to enzymatic degradation. This has indeed been realised in the development of meropenem; it possesses the broad spectrum of activity and resistance to beta-lactamases that are embodied in imipenem as well as displaying increased stability to human dehydropeptidases. Most recent research has focused upon the development of carbapenem antibiotics which combine broad spectrum antimicrobial activity and metabolic stability with oral absorption, for the treatment of community-acquired infections. Indeed, the pro-drug esters of the tricyclic carbapenems represent the first significant advance in this respect. However, the increased use of carbapenem antibiotics would undoubtedly accelerate the emergence of carbapenem-hydrolysing enzymes. The ultimate challenge could therefore be the design and synthesis of carbapenem derivatives that are resistant to these metallo-beta-lactamases. Due to the enormous problems encountered in the development of the carbapenem antibiotics, this area of research has, in the past, been described as a battlefield that did not bode well for the future [181]. Primaxin and meropenem proved however that these problems were not insurmountable, and are therefore a testimony to the persistence and dedication of those scientists in their war against