Phenotypic Characterization of Extended Spectrum Beta-Lactamase, Class C Cephalosporinase and Carbapenemase-Producing Klebsiella Species Isolated from Patients Consulted at Four Yaounde-Based Hospitals

Background: Klebsiella spp. are bacteria of medical importance for their role in opportunistic infections which are often difficult to treat because of resistance to one or several antimicrobials. The aim of this study was to determine antimicrobial resistance due to Extended Spectrum Beta-lactamase (ESBL), Class C cephalosporinase (AmpC) and carbapenemase enzymes in Klebsiella spp. isolated from patients consulted at four hospitals. Methodology: The study was cross-sectional and descriptive. A total of 4190 non-repetitive patients specimens from 13 types of clinical specimens were analysed from February to November 2020. Two hundred and twenty-five (225) Klebsiella spp isolates were identified using API 20E and antimicrobial susceptibility testing done according to the Kirby Bauer disc diffusion method. ESBL and AmpC phenotypes were determined by the combination disc method and carbapenemases by double disc synergy method, referenced by EUCAST guidelines for the resistance testing. Results: The frequency of the species was Klebsiella pneumoniae (69%, 155/255), K. oxytoca (14%, 31/255), K. ozaenae (12%, 27/225) and K. rhinoscleromatis (5%, 11/225). Isolates were most resistant to sulphomethoxazole trimethoprim (84%, 189/225), cepaholosporins (80%, 180/225), and least resistant to carbapenems (10.7%, 24/225). Two K. oxytoca and one K. pneumoniae were resistant to all antibiotics tested. Klebsiella pneumoniae had the most multidrug resistant isolates (59.4%, 134/225). Most isolates (83.6%, 188/225) expressed at least one enzyme, while 63.6% (143/225) of the isolates expressed at least two enzymes. Some isolates were ESBL (71.6%, 161/225), carbapenemase (10.7%, 24/225) and AmpC (6.6%, 15/225) producers. Three carbapenemases (Klebsiella pneumoniae carbapenemase-KPC, Metallo-Beta Lactamase-MBL and OXA-48) were detected. Conclusion: These results revealed that resistance of Klebsiella spp. to cephalosporins is high and this may be exacerbated by co-expression of AmpC and carbapenemases aggravating associated patient morbidity and mortality. Monitoring of antimicrobial resistance of local strains is necessary for informed decisions on empirical treatment. .

Occurrence of K1 and K2 serotypes and genotypic characteristics of extended spectrumβ-lactamases-producing Klebsiella pneumoniae isolated from selected hospitals in Malaysia

Objective:To determine the distribution,phenotypic and genetic background of extended spectrumβ-lactamases(ESBL)-producing Klebsiella(K.)pneumoniae clinical isolates associated with K1 and K2 serotypes in two selected hospitals in Malaysia.Methods:A total of 192 K.pneumoniae isolates were collected and subjected to antibiotic susceptibility,hypermucoviscosity test and multiplex PCR to detect the presence of K1-and K2-serotype associated genes.Multilocus sequence typing(MLST)was performed on ESBL-producing K.pneumoniae isolates presented with K1 and K2 serotypes,followed by phylogenetic analysis.Results:A total of 87 out of 192(45.3%)of the K.pneumoniae isolates collected were ESBL producers.However,only 8.3%(16/192)and 10.9%(21/192)of the total isolates were detected to carry K1-and K2-serotype associated genes,respectively.Statistical analysis showed that K1 and K2 capsular serotypes were not significantly associated with ESBL phenotype(P=0.196).However,they were significantly associated with hypervirulent,as demonstrated by the positive string test(P<0.001).MLST analysis revealed that ST23 as the predominant sequence type(ST)in the K1 serotype,while the ST in the K2 serotype is more diverse.Conclusions:Although the occurrence of ESBL-producing isolates among the hypervirulent strains was low,their coexistence warrants the need for continuous surveillance.MLST showed that these isolates were genetically heterogeneous.

Characteristics of β-Lactamase Synthesis in E. coli and K. pneumanie Strains in Nosocomial Infections

Background: Recently micro-organisms that synthesize extended-spectrum β-lactamase (ESBLs) were increased. The peculiarities of ESBL synthesis of Escherichia coli and Klebsiella pneumoniae strains that cause nosocomial urinary tract infections, surgical site infections and pneumonia in surgical clinic were studied. ESBL synthesis were observed 38.9% of E. coli strains obtained from urine, 92.3% of strains obtained from surgical site infections, and 50% of strains obtained from sputum. ESBL synthesis were observed 37.5% of K. pneumoniae strains obtained from urine, 85.7% of strains obtained from surgical site infections, and 60% of strains obtained from sputum. Different levels of ESBL synthesize of E. coli and K. pneumoniae strains isolated from different pattern is discussed. Conclusion. ESBL synthesis is common in E. coli and K. pneumoniae strains, which cause nosocomial infections. The frequency of occurrence of ESBL s synthesis among of these strains depends on clinical forms of nosocomial infections.

Analysis of AmpC β-lactamase Gene in Pseudomonas aeruginosa

The gene and the amino acid sequence of the structural and regulatory region of the Pseudomonas aeruginosa with different resistance patterns were analyzed. Six strains with different resistance patterns were selected and the AmpC β-lactamase was identified. The objective gene fragment was amplified by colonies PCR. The sequences of the PCR-products were analyzed. The DNA sequence of the structural gene ampC and the regulatory genes ampR, ampD and ampE was detected. The 6 strains and the wild-type Pseudomonas aeruginosa are highly homogeneous in structural and regulatory region. Some new mutant points were found.

Prevalence and Risk Factors of Penicillinase-Type β-Lactamase Producing Neisseria gonorrhoeae Isolated from Patients Attending Health-Facilities in Yaounde, Cameroon

Background and Objectives: Mitigation of antibiotic resistant Neisseria gonorrhoeae has become a priority due to considerable health and economical disabilities it generates. In order to tackle the emergence of resistant Neisseria gonorrhoeae, this study aimed to determine the prevalence and risk factors of penicillinase type β-lactamase-producing Neisseria gonorrheae among patients consulting for genital infectious disorders in two health-facilities in Yaounde, Cameroon. Materials and Method: A cross-sectional descriptive and analytical study was conducted over a 3-month period, from July 2nd to October 2nd, 2022. Vaginal and urethral secretions were collected. Biochemical identification tests were performed on colonies grown on chocolate agar + polyvitex using the Api NH gallery. The detection of penicillinases was equally performed using the API NH gallery and confirmed using the antimicrobial susceptibility testing. The Minimum Inhibitory Concentrations of some antibiotics were determined using the E-Test. Results: The results showed that out of the 198 patients sampled, 16 (8.08%) were positive for Neisseria gonorrhoeae, among which 13/16 (81.25%) were penicillinase-type β-lactamase producers. Antimicrobial susceptibility testing results showed high co-resistances to antibiotics, mainly ciprofloxacin (100%), nalidixic acid (92.31%) and azithromycin (84.62%). Moreover, high Minimum Inhibitory Concentrations of ceftriaxone (ranging from 6 to 24 mg/L) was observed toward Neisseria gonorrhoeae isolates. The risk factors of the carriage of penicillinase-type β-lactamase producing Neisseria gonorrhoeae identified were: a history of Sexually Transmitted infections (p = 0.01) and unprotected sexual intercourse (p = 0.01). Conclusion: The emergence of penicillinase-type β-lactamase producing Neisseria gonorrhoeae is increasing and the situation is becoming worrisome. The identified risk factors can constitute a basic outlook to tackle resistant Neisseria gonorrhoeae, and therefore sustain antibiotic stewardship.

Evaluation of Disk Potentiation Test (DPT) and Double Disk Synergy Test (DDST) for The Detection of Metallo-β-Lactamases (MBLs) in Clinical Isolates of Bangladesh

Objective: Increasing the emergence of Metallo-β-lactamase (MBL) producing gram-negative bacteria and their dexterous horizontal transmission demands rapid and accurate detection. This study was conducted to determine a suitable method to promptly detect MBL-producing gram-negative bacteria. Methods: A total of 103 gram-negative bacteria were identified from various clinical samples at a tertiary care hospital in Dhaka city. MBL producers were detected by two phenotypic methods, the Disk Potentiation Test (DPT) and the Double Disk Synergy Test (DDST) based on β-lactam chelator combinations where EDTA/SMA has been used as an inhibitor and Imipenem, Ceftazidime as substrates. Results: 103 isolates which were identified as Escherichia coli spp, Klebsiella spp, Pseudomonas spp, Acinetobacter spp, Proteus spp, Providencia spp were found to be multidrug-resistant in antibiogram test. Isolates showed complete resistance (100%) to Imipenem, Meropenem, and Amoxiclav. The highest carbapenem-resistant etiological agents were Acinetobacter spp 40 (38.8%) followed by Pseudomonas spp 27 (26.2%), Klebsiella spp 26 (25.2%), Escherichia coli 8 (7.8%), Proteus spp 1 (1%) and Providencia spp 1 (1%). DPT method detected significantly (p = 0.000009) a higher number of MBL-producers (Imipenem with 0.5 M EDTA n = 61, 59.2% & Ceftazidime with 0.5 M EDTA n = 56, 54.4%) compared to the DDST method (Imipenem -0.5 M EDTA n = 43, 41.7%, Imipenem – SMA n = 38, 36.9% & Ceftazidime -0.5 M EDTA n = 15, 14.6%). Conclusion: Pieces of evidence suggest that DPT is a more sensitive method than DDST and could be recommended for identifying MBL-producing bacteria in Bangladeshi hospitals for the proper management of patients, to reduce time constraints and treatment costs.

Multidrug resistance extended spectrum β-lactamase and AmpC producing Escherichia coli isolated from the environment of Bogor Slaughterhouse,Indonesia

To determine the multidrug resistance extended spectrum β-lactamase and AmpC (ESBL/AmpC producing) Escherichia coli (E. coli) isolated from the environment of Bogor slaughterhouse, Indonesia.MethodsA total of 35 samples from 7 locations in slaughterhouse i.e., source of water, slaughtering floor, swab of carcass area floor, swab of evisceration area floor, untreated waste water, treated waste water, drinking water for cattle were collected from March to April 2016. Presence of ESBL/AmpC producing E. coli and susceptibility testing against 8 antimicrobial agents (penicillin G, streptomycin, gentamycin, ciprofloxacin, enrofloxacin, tetracycline, trimethoprim-sulfamethoxazole, and polymyxin B) were detected by disk diffusion test according to Clinical and Laboratory Standards Institute.ResultsESBL/AmpC producing E. coli were identified in 14.3% (5/35) of the collected samples from the environment of Bogor slaughterhouse. ESBL/AmpC-producing E. coli isolates were detected in untreated waste water (n = 3), slaughtering floor (n = 1), and carcass area floor (n = 1). Most of ESBL/AmpC-producing E. coli isolates (80%) showed multidrug resistance phenotypes against at least three classes of antibiotics. The highest incidence of antibiotics resistance was against penicillin G (100.0%) and streptomycin (100.0%), followed by gentamicin (60.0%), trimethoprim-sulfamethoxazole (60.0%), tetracycline (40.0%), ciprofloxacin (40.0%), enrofloxacin (20.0%), and polymyxin B (0.0%).ConclusionsThe transmission of antimicrobial resistant bacteria into the environment may be a potential risk for human health.

Antimicrobial Susceptibility and Genotyping of Plasmid Mediated- Extended Spectrum β-lactamase, AmpC β-lactamase and Aminoglycoside Modifying Enzymes in Klebsiella pneumoniae

The antimicrobial susceptibility and genotyping of plasmid mediated-extended spectrum β-lactamase (ESBLs), AmpC β-lactamase (pAmpC) and aminoglycoside modifying enzymes (AMES) in Klebsiella pneumoniae isolated in Tianjin, China were investigated in the present study. Sixty strains of Klebsiella pneumoniae isolated from Huanhu Hospital were tested against 21 antimicrobial agents by Microscan MIC method, and the PCR and DNA sequencing were used to determine the genotypes of ESBLs, pAmpC and AMES. The result in antimicrobial susceptibility testing showed that all the 60 strains of bacteria tested were proved to be multi-resistant, and that of genotyping demonstrated different genotypes among these bacterial strains, in which 60 strains belonged to the TEM type; 25 strains to the CTX-M type and 54 strains to the DNA pAmpC type. The AMES genes were found in 59 strains, while the CTX-M type of ESBLs, DHA pAmpC type AMES genes were detected in 22 strains simultaneously. It concludes that the problem of drug-resistance of Klebsiella pneumoniae on hospital in Tianjin is a serious issue, and at least 3 kinds of β-lactamase and 3 AMES genes exist.

L1 β-Lactamase催化反应机理研究

用混合量子力学和分子力学(QM/MM)方法和密度泛函理论讨论了L1β-Lactamase催化Nitrocefin水解的过程,研究结果表明,反应为多步反应:第一步亲核进攻反应为反应的决速步骤,并且伴随着酰胺键的断裂,第二步反应为质子迁移反应.同时讨论了金属锌在反应中的作用.

Prevalence and characteristics of extended spectrum β-lactamase-producing Escherichia coli from bovine mastitis cases in China

The aim of the study was to investigate the prevalence and characterization of extended-spectrum β-lactamase （ESBL）- producing Escherichia coli isolated from bovine mastitis cases in China. ChromID ESBL agar was used to confirm ESBL-producing E. coli. PCR and DNA sequencing were employed to characterize the genotype of ESBL-producers. Antimicrobial susceptibility was measured by disc diffusion. Overall, 73 of 318 E. coli isolates （22.96％） were identified as ESBL-producers. Of these ESBL-producing E. coli, the prevalence of blaCTX-M and blaTEM-1 was 97.26 and 71.23％, respectively. The predominant CTX-M-type ESBL was CTX-M-15 （65.75％）, followed by CTX-M-14 （10.96％）, CTX-M-55 （9.59％）, CTX-M-64 （5.48％）, CTX-M-65 （4.11％） and CTX-M-3 （1.37％）. This study is the first report of CTX-M-64 and CTX-M-65 in E. coli isolated from bovine mastitis. Furthermore, 72 ESBL-producing E. coli isolates （98.63％） were found to be multidrug-resistance. This study noted high prevalence and rates of antimicrobial resistance of ESBL-producing E. coli isolates from bovine mastitis cases in China.

Extended-spectrum β-lactamase controversies

G.A.Jacoby 教授毕业于美国哈佛医学院,为国际知名的传染病学、微生物学和免疫学专家,曾长期担任美国麻省总医院传染病科医师,现任麻省 Lahey 医学中心传染病实验室主任,中国感染与化疗杂志特邀编委。长期以来 Jacoby 教授对细菌耐药机制有深入研究,尤其对细菌(肺炎克雷伯菌、大肠埃希菌等)产生质粒介导的超广谱β内酰胺酶(ESBL)以及喹诺酮类的耐药机制研究。曾发表多篇有关上述专题的权威性论著。此次本刊特邀 Jacoby 教授撰写有关 ESBL 的新进展,文中阐述目前 ESBLs 除通常所指质粒介导β内酰胺酶中扩大了酶水解的底物谱导致细菌对头孢噻肟、头孢他啶、氨曲南等抗生素耐药外,还包括许多具有不同特点的β内酰胺酶,其中某些酶产自共生菌。除 ESBL 外,AmpC 酶和各种碳青霉烯β内酰胺酶也可导致细菌对上述抗生素耐药,因此临床微生物实验室准确检出和鉴别各种β内酰胺酶,对于临床选用适宜的抗茵药十分重要。美国 CLSI(过去称为 NCCLS)推荐采用两步法(筛选及确证)检测细菌产 ESBL;但有的学者认为测定抗茵药物对细菌的 MIC,如用药后 T>MIC 在50%以上即可达到满意疗效,无需检测细菌是否产 ESBL。目前认为碳青霉烯类抗生素治疗产 ESBL 菌感染的疗效最为满意,但由此可能引起细菌对该类药物耐药,值得关注。采用大剂量头孢吡肟或哌拉西林-三唑巴坦治疗产ESBL 菌感染是否有效尚有争论,临床上对β内酰胺类最为耐药的菌株往往对几乎所有现用抗菌药耐药。黏菌素(或多黏菌素 B)曾成功的用于治疗此种多重耐药菌感染。此外替莫西林(temocillin,一种对β内酰胺酶稳定的青霉素类)与替加环素(tigecvcline,为米诺环素衍生物)体外对产 ESBL 菌有抗菌作用,但尚无临床研究资料。

Antimicrobial Resistance and Genotype Analysis of Extended-Spectrum-β-Lactamase-Producing Proteus Mirabilis

To analyse the genotypes of clinical isolates of Extended-Spectrum-β-Lactamase-Producing (ESBL-producing) Proteus mirabilis (P. mirabilis) and the mechanisms of antimicrobial resistance, to guide reasonable use of antibiotics and to avoid nosocomial outbreak infections by ESBL-producing P. mirabilis. 125 clinical isolates of P. mirabilis were collected from the Drug-Resistant Bacteria Surveillance Center of Anhui Province (from Jan 2009 to May 2010). Searching for the genotypes of ESBLs was perfomed by PCR amplification and DNA sequencing, and performed conjugation test simultaneously. Among ESBL-producing strains, CTX-M was the major genotype (3 CTX-M-13 and 1 CTX-M-3). TEM-1b spectrum β-lactamase was also prevalence in P. mirabilis. The diversity of β-lactamases in P. mirabilis and the emergency of multi-drug-resistance clinical strains will present serious threat to clinical therapy and even will lead to outbreak of nosocomial infections. Our study emphasizes the need for enhanced supervision of ESBL-producing P. mirabilis. Timely and reasonable drug-resistance data are indispensable to clinical therapy.

产AmpC酶的猪源奇异变形杆菌分离鉴定及生物学特性分析

奇异变形杆菌(Proteus mirabilis)是一种可导致多种动物感染的条件致病菌,目前有关产AmpC酶的动物源奇异变形杆菌研究较少。为探究广西某猪场母猪流产、死胎的原因,从流产猪胎肝脏中分离到病原菌,对分离株进行纯化培养、染色镜检、生化试验和16S rRNA测序确定为奇异变形杆菌,命名为GX-Y9251株。对GX-Y9251株进行药敏试验、耐药基因检测、AmpC酶基因检测、毒力基因检测、小鼠致病性试验和生物膜半定量检测。药敏试验结果显示,该菌株对8种药物敏感,对17种抗生素耐药。GX-Y9251株存在多种耐药基因,且产AmpC酶,为blaDHA基因。GX-Y9251株存在6个毒力基因,其对小鼠的半数致死量(LD50)为5.4×107CFU;具有生物被膜形成能力,为中等黏附型(++)。研究结果为奇异变形杆菌感染的防治提供了理论依据。

Comparative study on occurrence of class A and class Cβ-lactamase genes and their co-occurrence in Indian Enterobacteriaceae during years 2009 and 2010

Objective:To determine the occurrence of class A and class Cβ-Iactamase genes and their cooccurrence in Indian Enterobacteriaceae.Methods:52 third generation cephalosporin resistant isolates were phenotypically detected by combination disk method and screened by PCR to identify class A and class C typeβ-lactamase genes.Results:Of the 52 isolates,94.2%（49） were found harboring any of the bla?,blaCTX-M,blaSHV and blaTEM were present in 82.6%（43/52）, 59.6%（31/52）,and 42.3%（22/52） isolates,respectively.Of the 49 ESBL positive isolates 57.1% （28/49） showed co-occurrence of blaampCwith bla?.On the contrary,the collection from 2009 showed their co-occurrence in 81.4%isolates.Conclusions:The comparative study shows a downward trend for co-existence of bla? with blaampC from 2009 to 2010.Further large scale studies are needed to address the co-occurrence of class A and class Cβ-lactamases in India and the resistance trend occurring over a period of time.

EXTRACELLULAR RELEASE OF β-LACTAMASE BY OSMOTIC SHOCK IN SYNECHOCOCCUS TRANSFORMANT

A unicellular cyanobacterium Synechococcus sp. strain PCC 7002 was transformedwith plasmid pQL1, on which β-lactamase gene (bla) and β-galactosidase gene (lacZ) were encoded.The transformant cells released β-lactamase into medium by an abrupt drop of osmotic pressure. This re-sult indicates that this cyanobacterium recognizes and processes the signal sequence of β-lactamase, andaccumulates the enzyme in periplasm. Repeated release of β-lactamase was possible by repeated osmoticshocks without impairing cell viability. On the other hand, most of the β-galactosidase remained in cyto-plasm under the osmotic shock.

Metallo-β-lactamase producing nonfermentative gram-negative bacteria:An increasing clinical threat among hospitalized patients

Objective:To detect and evaluate the various methods for metallo-β-lactamases(MBL) production in Pseudomonas aeruginosa(P.aeruginosa) and Acinetobacter species.Methods:A total of 109 P.aeruginosa and 85 Acinetobacter species were screened for imipenem resistance by Kirby- Bauer disc diffusion methods.Detection of MBL production was(lone by imipenem-EDTA combined disc test,double disc synerygy test(DDST) and imipenem-EDTA MBL E test.Results: A total of 63(57.8%) strains of P.aeruginosa and 46(54.1%) strains of Acinetobacter spp.were found to be resistant to imipenem.Of the 63 imipenem resistant P.aeruginosa tested for MBL production.44(69.89;) were found to be positive and among 46 imipenem resistant Acinetobacter. 19(41.3%) were shown to be the MBL producers.Conclusions:Imipenem-EDTA combined disc test and MBL E test are equally effective for MBL detection in both P.aeruginosa and Acinetobacter spp.,but given the cost-constraints,combined disc can be used as a convenient screening method in the clinical microbiology laboratory.

Studies of β-lactamase production in ampicillin resistant Haemophilus influenzae

Objective:To study the molecular mechanisms of β-lactamase production in ampicillin resistant(AmP ) Hae- mophilus influenzae(HI). Methods: Identified the β-lactamases production strain from AmP HI was isolated from clinical cases with K-B method. β-lactamase encoding gene in enzyme production strains were detected by PCR with lactamase gene specific primers, and both plasmid and chromosomal DNA samples. Results: Thirty-two out of 36 (88 .9% ) were found to be β-lactamase production. Twenty-nine out of 32 enzyme production stain were PCR positive (the ratio of PCR positives 90.6% ). There were 25 stains amplified with plasmid DNA positively, and 4 with chromosomal DNA. Conclusion: (l ) Most of the AmPr HI strain produce lactamase is mediated by plasmid. (2) Detection of lactamase encoding gene in HI is a simple and efficient approach to study the molecular basis of ampicillin resistance.

Intestinal carriage of methicillin resistant Staphylococcus aureus and extended-spectrum β-Lactamase-producing Enterobacteriacae in hospitalized and nonhospitalized patients and their clinical implications

Objective:To determine the clinical implication of and intestinal carriage with methicillin resistant Staphylococcus aureus(MRSA) and extended spectrumβ-lactamase(ESBL)-producing Enterobacteriacae.Methods: A total of 180 stool specimens were screened for MRSA and ESBL-producing enterobacteria.Identification of ESBL- producing Enterobacteriacae was done by MicroScan Walk Away 96 system(Dade Behring Inc.,West Sacramento,CA 95691,USA ) and confirmation by double-disc synergy test.MRSA was identified by disc diffusion using 30μg cefoxitin disc and the MicroScan.Results:The rate of fecal MRSA carriage was 7.8% (14/180),35.7%(5 /14) were recovered from surgical wards.Three patients(21,4%) had MRSA recovered from other body sites,and 2(14.2%) had in addition ESBL -producing Escherichia coli(E.coli) and Klebsiella pneumoniae(K.pneumoniae) respectively.Four(28.5%) patients with MRSA fical carriage died. MRSA fecal carriage was recovered from both inpatients and outpatients.Four(2.2%) cases carried ESBL-producing Enterobacteriacae in feces.Three(75%) were from intensive care unit(ICU).One patient had both ESBL-producing E.coli and K.pneumoniae from stool as well as E.coli from tracheal aspirate.Two ICU patients with fecal ESBL died.Conclusion:Fecal screening for MRSA and ESBL of all patients at high risk admitted to different hospital wards and ICUs and implementing infection control measures were recommended.

Metallo-β-Lactamases:A Major Threat to Human Health

Antibiotic resistance is one of the most significant challenges facing global healthcare. Since the 1940s, antibiotics have been used to fight infections, initially with penicillin and subsequently with various derivatives including cephalosporins, carbapenams and monobactams. A common characteristic of these antibiotics is the four-memberedβ-lactam ring. Alarmingly, in recent years an increasing number of bacteria have become resistant to these antibiotics. A major strategy employed by these pathogens is to use Zn(II)-dependent enzymes, the metallo-β-lactamases (MBLs), which hydrolyse theβ-lactam ring. Clinically useful MBL inhibitors are not yet available. Consequently, MBLs remain a major threat to human health. In this review biochemical properties of MBLs are discussed, focusing in particular on the interactions between the enzymes and the functionally essential metal ions. The precise role(s) of these metal ions is still debated and may differ between different MBLs. However, since they are required for catalysis, their binding site may present an alternative target for inhibitor design.

Microbiologic and Clinical Comparison of Patients Harboring <i>Escherichia coli</i>Blood Isolates with and without Extended-Spectrum <i>β</i>-Lactamases

The clinical and microbiologic characteristics of 34 patients with extended-spectrum β-lactamase (ESBL) positive E. coli isolated from blood were compared to 66 bacteremic patients with ESBL negative E. coli, from January 2007 through December 2009. Of the 21 ESBL positive isolates available for PCR analysis, 13 were positive for CTX-M, 8 for TEM, 4 for SHV β-lactamases, with 6 possessing multiple enzymes. Twenty of 34 (59%) ESBL-positive and 41 of 66 (62%) ESBL-negative blood isolates were considered community-associated. All but one isolate in both groups had MICs of ≤1.0 μg/ml to meropenem. However, when compared to ESBL-negative isolates, ESBL-positive isolates were more frequently resistant to levofloxacin, trimethoprim/sulfamethoxazole and had higher MICs to gentamicin, tobramycin and piperacillin/tazobactam. The use of intravenous and urinary catheters was strongly associated with the isolation of E. coli bloodstream isolates in both groups of patients. Although hospital stay was similar in both groups, appropriate therapy was given in 87% of patients with ESBL positive vs. 98% of patients with ESBL negative isolates and mortality was greater for patients with ESBL positive isolates (26% vs. 17%). Since a large proportion of E. coli blood isolates were ESBL-positive and community-associated, carbapenems should be considered as initial empiric therapy for such infections in our locale.