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Prevalence of extended-spectrum beta-lactamases (ESBLs) produced in blood isolates of gram-negative bacteria in a teaching hospital in southern Thailand.

One hundred and eighty-three strains of gram-negative bacteria were isolated from 177 bacteremic patients during a 6-month period in 2004 at a teaching hospital in southern Thailand. Extended-spectrum beta-lactamases (ESBL) production was detected in K. pneumoniae, E. coli, and E. cloacae, at rates of 16/36 (44.4%), 3/59 (5.1%), and 2/13 (15.4%), respectively. All but one of the screened positive strains were also positive by both the combination disk method and the E-test. All of the K. pneumoniae strains that were resistant to ceftazidime by disk diffusion were demonstrated to be positive for ESBL production by both the combination disk and E-test methods. Most of the ESBL positive strains had a high MIC (more than 32 microg/ml) to ceftazidime. However, all the ESBL positive strains were sensitive to imipenem.

Anti-Bacterial Agents↗

[Occurrence of extended spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) in clinical strains of gram-negative rods].

This study was undertaken to check the situation concerning the occurrence of Gram-negative rods producing extended-spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) in clinical specimens from patients hospitalized in National Clinical Hospital No. 1 in Warsaw. Such determinations were not performed in this hospital so far. During three months (April-June, 1997) 200 strains of Gram-negative rods were cultured. The strains were identified in automatic ATB system using strips with biochemical tests: ID 32 E for enteric rods and ID 32 GN for non-fermenting rods. ESBL-producing strains were detected with double disc diffusion test according to Jarlier et al. (1988). Clavulanate was applied as the inhibitor of beta-lactamases (AMO/CLAV disc). Inducible beta-lactamases were determined using double disc method according to Sanders and Sanders (1979). Cefoxitin was the inductor of these beta-lactamases. 82 strains (41% of all strains) belonging to Enterobacteriaceae family, 92 strains (46%) of Pseudomonadaceae rods and 26 strains (13%) of other Gram-negative rods were isolated. 30 ESBL-producing strains (15% of all strains) and 45 strains (22.5%) with IBL activity were detected. The obtained results confirm the necessity of continuous and reliable monitoring of ESBL--and IBL--producing strains among Gram-negative rods isolated from clinical materials. The aims of such procedure are the control and prevention of their dissemination within a hospital as well as the avoidance of therapeutic failures.

Clavulanic Acid↗

Investigating the zoonotic origins of ESBL-producing E. coli in community-acquired urinary tract infections in Ecuador.

Extended-spectrum β-lactamase-producing Escherichia coli (ESBL-producing E. coli) pose a growing global health threat. Although Latin America has been identified as a global hotspot of antimicrobial resistance, the zoonotic contribution to drug-resistant infections in the region remains poorly defined. We analyzed 137 clinical ESBL-producing E. coli isolates from urinary tract infections (UTIs) in Quito, Ecuador, applying a Bayesian latent class model informed by host-associated mobile genetic elements to estimate the fraction of infections attributable to food-animal sources. We estimated that 25.5% (35/137) of UTI isolates were putative zoonotic cases. This proportion rose to 42.5% after excluding ST131-H30, a human-associated pandemic lineage. Putative zoonotic isolates were enriched for animal-associated β-lactamase genes (e.g., blaTEM-1B, blaCTX-M-65), lacked human-associated markers such as blaOXA-1, and exhibited diverse antimicrobial resistance gene profiles resembling those observed among food-animal isolates. These isolates were also enriched for ColV-associated virulence genes typically linked to avian pathogenic E. coli. Putative zoonotic strains contributed substantially to third-generation cephalosporin-resistant UTIs in Quito, Ecuador, challenging assumptions derived from high-income settings that such infections are driven predominantly by human-to-human transmission. These findings highlight the importance of integrated One Health surveillance and mitigation, particularly in low- and middle-income countries where gaps in water, sanitation, and hygiene (WASH) may interact with antimicrobial use in food production to amplify antimicrobial resistance transmission.IMPORTANCEESBL-producing E. coli have rapidly emerged as a major global antimicrobial resistance threat. In Latin America, cephalosporins are commonly used in food-animal production, fueling the emergence of ESBL-producing E. coli. In low- and middle-income countries, excessive antimicrobial use driven by poorly regulated over-the-counter sales, combined with inadequate water, sanitation, and hygiene (WASH) infrastructure, can facilitate antimicrobial-resistant pathogen transmission from food animals to humans. Using a novel statistical-genomic approach, we found that over one in four cephalosporin-resistant UTIs in Quito, Ecuador, may be caused by E. coli strains originating from food animals. Our findings highlight the public health risks associated with antimicrobial use in food-animal production and the role of environmental and infrastructure-related vulnerabilities. As global demand for animal protein continues rising in middle-income countries, controlling zoonotic antimicrobial resistance transmission becomes increasingly urgent for protecting human health through integrated One Health strategies.

ESBL-producing E. coli↗

ESBL- and plasmidic class C beta-lactamase-producing E. coli strains isolated from poultry, pig and rabbit farms.

This study aims to determine the presence of extended-spectrum (ESBL) and plasmidic class C beta-lactamase-producing Enterobacteriaceae in poultry, pig and rabbit farms of Catalonia (Spain). PFGE typing showed a low clonal relationship among strains carrying these mechanisms of resistance. Ninety-three percent of them were resistant to two or more of the non-beta-lactam antimicrobials tested and harboured ESBL and plasmidic class C beta-lactamases. Greater diversity of these enzymes was found in strains from poultry farms, the CTX-M-9 family, especially CTX-M-14, with CMY-2 being the most frequent. The isolation of TEM-52 and SHV-2-producing Escherichia coli strains from these animal farms is noteworthy. In contrast, 73% of the strains from pig farms had CTX-M-1, and neither the CMY-type nor CTX-M-9 family enzyme was found. Likewise, it is the first time that CTX-M-1 and SHV-5 encoding strains have been isolated in pigs. On the other hand, in rabbit farms CTX-M-9 family was also the most frequent, being detected in three of a total of four strains. The last one showed a CMY-2, for the first time detected in these animals, too. In conclusion, commensal E. coli strains of food-producing animal farms are a reservoir of ESBL and plasmidic class C beta-lactamases.

Animals↗

The synthesis of 7-substituted-3-dinitrostyryl cephalosporins and their ability for detecting extended spectrum beta-lactamases (ESBLs).

We synthesized 7-substituted-3-(2,4-dinitrostyryl)cephalosporin derivatives which were Nitrocefin analogs, for detecting extended spectrum beta-lactamase (ESBL) specifically. HMRZ-86 which has carboxypropyloxyimino group on 7-aminothiazolacetamide substituent were not hydrolyzed by class A, C and D beta-lactamases, but it was hydrolyzed by ESBL and metallo beta-lactamase (class B), then its color changed from yellow to red. The hydrolysis of metallo beta-lactamase was inhibited by adding sodium mercapto acetic acid (SMA). Therefore HMRZ-86 is a useful chromogenic agent to detect ESBL specifically.

Cephalosporins↗

[Extended spectrum beta-lactamse (ESBL)-induced antibiotics resistance in gram-negative agents: what should be watched in intensive care medicine?].

Third-generation cephalosporins have an important part in the therapy of infections due to gram-negative bacteria. Extended spectrum beta-lactamases (ESBL) are able to hydrolize these antibiotics. As the number of ESBL-producing bacteria is increasing worldwide, detection of ESBL-producers is important for the both of prevention and therapy.

Critical Care↗

Ceftazidime-hydrolysing CTX-M-15 extended-spectrum beta-lactamase (ESBL) in Poland.

The CTX-M-15 extended-spectrum beta-lactamase (ESBL) was recently identified in Enterobacteriaceae isolates in India, and demonstrated significant hydrolytic activity against ceftazidime, in contrast to the majority of CTX-M enzymes. CTX-M-15 differs from CTX-M-3, which is one of the most prevalent ESBLs in Poland, by only a single amino acid change (Asp-240-->Gly). Three cefotaxime- and ceftazidime-resistant Enterobacteriaceae isolates, recovered during 1998-2000 in two Polish hospitals, were found to produce CTX-M-15. Similar to those from India, the isolates contained the ISEcp1 insertion sequence located upstream of the bla(CTX-M-15) gene, which has been recently demonstrated to mobilize 3'-adjacent genes to transfer between DNA replicons. However, its different position with respect to the beta-lactamase gene indicated the independent selection of the ESBL gene in the two countries.

Anti-Bacterial Agents↗

CTX-M: changing the face of ESBLs in Europe.

Since around 2000 - earlier in Poland and Spain and later in France and the UK - dramatic shifts have occurred in the prevalence and types of extended-spectrum beta-lactamases (ESBLs) in Europe. Before this watershed, most producers were nosocomial isolates, often Klebsiella spp. or Enterobacter spp. from specialist care units, and had mutant TEM or SHV ESBLs. Subsequently, CTX-M ESBLs have become dominant, with much greater penetration into Escherichia coli, and with many infections in 'complicated community' patients, usually with underlying disease, recent antibiotic usage, or healthcare contact. The degree of clonality among producers varies with the country, as does the enzyme type produced, with group 9 (CTX-M-9 and -14) enzymes dominant in Spain and group 1 enzymes (particularly CTX-M-3 and -15) dominant elsewhere. Irrespective of the particular enzyme, most producers are multiresistant. These changing patterns present major therapeutic and infection control challenges, with the public health intervention points unclear.

Community-Acquired Infections↗

High-dose cefepime as an alternative treatment for infections caused by TEM-24 ESBL-producing Enterobacter aerogenes in severely-ill patients.

This study evaluated retrospectively the efficacy of treatment with cefepime vs. a carbapenem, in combination with amikacin or ciprofloxacin, for seriously-ill patients infected with ESBL-producing Enterobacter aerogenes who were admitted to an intensive care unit. Forty-four episodes of infection were investigated in 43 patients: 21 treated with cefepime; 23 with a carbapenem. The two treatment groups did not differ statistically in terms of age, APACHE II scores, and infection sites, but the average duration of antibiotic exposure was significantly shorter in the cefepime group (8.5 days vs. 11.4 days; p 0.04). Clinical improvement was seen in 62% of patients receiving cefepime vs. 70% of patients receiving a carbapenem (p 0.59). Bacteriological eradication was achieved in 14% of patients receiving cefepime vs. 22% of patients receiving a carbapenem (p 0.76). The 30-day mortality rates related to infection were 33% in the cefepime group and 26% in the carbapenem group (p 0.44). Thus, outcome parameters did not differ significantly between the two groups. Nevertheless, a statistically significant increase in failure to eradicate ESBL-producing E. aerogenes was observed as the MICs of cefepime rose (p 0.017). Pulsed-field gel electrophoresis revealed three distinct clones, but one predominant clone harbouring the bla(TEM-24) gene was associated with most (42/44) of the episodes of infection. It was concluded that cefepime may be an alternative agent for therapy of severe infections caused by TEM-24 ESBL-producing E. aerogenes, although further studies are required to confirm these observations.

Adult↗

ESBL - from petri dish to the patient.

Treatment of extended spectrum beta-lactamase (ESBL) producing strains of Enterobacteriaceae has emerged as a major challenge in hospitalised as well as community based patients. Infections due to ESBL producers range from uncomplicated urinary tract infection to life threatening sepsis. Although several reviews have been published in the literature about the detection and identification of these pathogens in the laboratory, there is not much guidance in Indian literature about how these organisms should be treated in clinical settings. The present article tries to address the clinical questions in the management of ESBL producing organisms. The article emphasises on antibiotic choice, pharmaco-therapeutic considerations, non-antibiotic aspects of management, and testing of clinical samples in the initial screening of patients for resistant gram negative organisms.

Anti-Bacterial Agents↗

[Detection methods for drug-resistant bacteria in routine examination--ESBL].

Extended-spectrum beta-lactamases(ESBLs) confer resistance to cefotaxime, ceftazidime, aztreonam, extended-spectrum penicillins, and structurally related beta-lactams in clinical isolates of K. pneumoniae and E. coli. Some ESBLs, however, show a lower level of resistance and these isolates may not reach NCCLS breakpoints for resistance. Optimal methods for confirmation of ESBL-producing isolates are described.

Escherichia coli↗

[Occurrence of beta-lactamase types ESBL and AmpC among gram-negative rods isolated from food].

The aim of the study was to isolate extended spectrum beta-laktamases (ESBL) and chromosomal beta-laktamases AmpC producing strains from food products. A total of 739 Gram-negative bacteria were tested with double disc diffusion method using cefotaxim, ceftasidim and amoxycillin with clavulanic acid. One strain producing ESBL and belonging to Stenotrophomonas maltophilia was detected from the ice cream (0.14% of all strains). From different food products a total of 14 microorganisms (1.9%) having AmpC enzymes have been isolated. They belonged to Enterobacter spp, Hafnia spp, Morganella spp, Citrobacter, Acinetobacter, and Cedecea. Appearance of strains producing beta-laktamases ESBL and AmpC among microorganism isolated from food make it necessary to monitor enzymes activity during routine microbiological control of foods.

Bacterial Proteins↗

[Genotyping of EsBLs-producing Escherichia coli by pulsed-field gel electrophoresis in nosocomial infection].

OBJECTIVE: Using a genotying method which is accurate and rapid to study the molecular epidemiology of EsBLs-producing Escherichia coli in nosocomial infection. METHODS: After an analysis of clinical information to ascertain nosocomial infection, EsBLs were detected by phenotype confirmation test and pulsed-field gel electrophoresis (PFGE). RESULTS: Three EsBLs-producing strains in ICU were verified as epidemiological-related because their dice coefficient index was higher than 95%. The others were verified as non-epidemiological-related since their different lanes were more than 6. CONCLUSION: PFGE is a perfect molecular typing method to study epidemiolgy and analyze homology.

China↗

[Detection of the expression of extended-spectrum beta-lactamases (ESBL) in clinical strains of Klebsiella pneumoniae and Escherichia coli: comparison of E-test and two-disc methods].

The aim of this study was to compare the effectiveness of E-test and two disc methods applied for the detection of extended spectrum beta-lactamases. All strains were tested by E-test, by double-disc synergy test (DDST) according to Jarlier (cefotaxim, ceftazidim, aztreonam and clavulanic acid) and also by disc test according to Appleton (cefpodoxime and cefpodoxime with clavulanic acid, CPD and CD01 disc). We tested 148 clinical strains of E. coli and 78 strains of K. Pneumoniae. In case of K. pneumoniae, the activity of the ESBLs was detected among 30 strains--both in E-test, Jarlier test and Appleton test. Among E. coli, four strains were found ESBL-positive in the test according to Jarlier but only three strain of these when E-test and Appleton test was used. The results of investigations performed suggest, that E-test and disc methods according both Jarlier and Appleton have the same effectiveness in detection ESBLs among K. pneumoniae strains. However, in case of E. coli, interpretation of results may present a problem.

Escherichia coli↗

[Antibiotic susceptibility and occurrence of ESBL in Pseudomonas aeruginosa strains isolated from different clinical specimens].

The aim of this study was to evaluate the drug susceptibility of 132 P. aeruginosa strains isolated from patients hospitalized in SPSK University Hospital in Bialystok. The isolates were obtained from clinical specimens over an 11-month period in 2001 and 2002. All the strains were identified in automatic ATB system using API 20 NE strips, and their susceptibility to antibiotics was tested by standard disc-diffusion method and agar dilution method. The minimal inhibitory concentration (MIC) was determined for five antibiotics: piperacillin, amikacin, ceftazidime, imipenem and ciprofloxacin. The majority of strains were susceptible to ceftazidime (91.7%), piperacillin combined with tazobactam (85.6%), amikacin (80.3%), meropenem and imipenem (81.8%). Many of our strains were resistant to cefotaxime (73.5%), ticarcillin (53%) and ciprofloxacin (48.5%). Also, the trial was undertaken to detect strains producing extended-spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) among P. aeruginosa rods isolated from different specimens. ESBL-producing strains were detected with double disc test (DDST) and combination double disc (CD) test. Clavulanate was applied as the inhibitor of these beta-lactamases. Strains producing ESBL were not found. On the other hand, as many as 127 P. aeruginosa strains (96.2%) produced inducible beta-lactamases (IBL).

Amikacin↗

[Conjugative transfer frequency of resistance genes from ESBL-producing Enterobacteriaceae strains isolated from patients hospitalized in pediatric wards].

The aim of this study was to evaluate the transfer frequency of plasmids encoding extended-spectrum beta-lactamases (ESBLs) from clinical isolates of Enterobacteriaceae to E. coli K12 C600 recipient strain. Additionally, resistance patterns to antimicrobial drugs of the isolates as well as transconjugants were analyzed. Fifty-four clinical strains belonging to the Enterobacteriaceae family were isolated from children hospitalized in Medical University Hospital in Wrocław. All the strains studied were identified in automatic ATB system using ID32E tests. Besides, they were ESBL-positive as was confirmed by the double-disc synergy test (DDST). The minimal inhibitory concentration (MIC) was determined for twelve selected antibiotics and chemotherapeutics. The majority of the strains (87%) were able to transfer plasmid-mediated ESBL to E. coli K12 C600 recipient strain with a frequencies ranged from 10(-5) to 10(-1) per donor cell. All the isolates studied as well as their transconjugants were susceptible to imipenem, meropenem and norfloxacin (MIC <1mg/L). On the other hand, these strains displayed high level of resistance (MIC 512 - >1024 mg/L) to cefotaxime, ceftriaxone, gentamycin, amikacin and cotrimoxazole. Genetic markers conferring resistance to aminoglycosides and cotrimoxazole were often co-transferred to recipient strain in conjugation process.

Anti-Bacterial Agents↗

Faecal prevalence of extended-spectrum Beta-lactamase (ESBL)-producing coliforms in a geriatric population and among haematology patients.

Antimicrobial resistance to the extended-spectrum cephalosporins is increasingly reported worldwide. In the local setting, nosocomial infections with multi-resistant Gram-negative bacilli are not uncommon and are a growing concern. However, there is limited data on the carriage rates of such organisms in the local setting. In May 2001, a prospective study was carried out to determine the enteric carriage rates of ceftazidime-resistant Gram negative bacilli (CAZ-R GNB) among residents of nursing homes and from in-patients of the geriatric and adult haematology wards of University Malaya Medical Centre. Ceftazidime-resistant Gram-negative bacilli (CAZ-R GNB) were detected in 25 samples (30%), out of which 6 were from nursing home residents, 5 from geriatric in-patients and 14 from the haematology unit. A total of 28 CAZ-R GNB were isolated and Escherichia coli (10) and Klebsiella pneumoniae (7) were the predominant organisms. Resistance to ceftazidime in E. coli and Klebsiella was mediated by extended-spectrum beta-lactamases (ESBLs). Although the majority of the CAZ-R GNB were from patients in the haematology ward, the six nursing home residents with CAZ-R GNB were enteric carriers of ESBL-producing coliforms. Prior exposure to antibiotics was associated with carriage of ESBL organisms and to a lesser extent, the presence of urinary catheters.

Aged↗

Identification of vaccine candidate antigens of an ESBL producing Klebsiella pneumoniae clinical strain by immunoproteome analysis.

Klebsiella pneumoniae is an opportunistic pathogen which causes pneumoniae, urinary tract infections and septicemia in immunocompromised patients. Hospital outbreaks of multidrug-resistant K. pneumoniae, especially those in neonatal wards, are often caused by strains producing the extended-spectrum-beta-lactamases (ESBLs). An immunoproteome based approach was developed to identify candidate antigens of K. pneumoniae for vaccine development. Sera from patients with acute K. pneumoniae infections (n = 55) and a control group of sera from healthy individuals (n = 15) were analyzed for reactivity by Western blot against ESBL K. pneumoniae outer membrane proteins separated by 2-DE. Twenty highly immunogenic protein spots were identified by immunoproteomic analysis. The immunogenic proteins that are most frequently recognized by positive K. pneumoniae sera were OmpA, OmpK36, FepA, OmpK17, OmpW, Colicin I receptor protein and three novel proteins. Two of the vaccine candidate genes, OmpA (Struve et al. Microbiology 2003, 149, 167-176) and FepA (Lai, Y. C. et al.. Infect Immun 2001, 69, 7140-7145), have recently been shown to be essential in colonization and infection in an in vivo mouse model. Hence, these two immunogenic proteins could serve as potential vaccine candidates.

Acute Disease↗