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The primary aim of this study was to collect national epidemiological data on candidaemia and to determine the reporting time of species identification and antifungal susceptibility in clinical practice. During a 1-year period (March 2013 until February 2014), every first Candida isolate from each episode of candidaemia was included prospectively from 30 Belgian hospitals. Identification and susceptibility testing were performed according to local procedures and isolates were sent to the National Reference Center for Mycosis. Species identification was checked by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and internal transcribed spacer (ITS) sequencing in case no reliable identification was obtained by MALDI-TOF MS. Antifungal susceptibility testing was performed according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology. A total of 355 isolates were retrieved from 338 patients. The mean incidence rate of candidaemia was 0.44 (range: 0.07 to 1.43) per 1000 admissions or 0.65 (range: 0.11 to 2.00) per 10,000 patient days. Candida albicans was most frequently found (50.4 %), followed by C. glabrata (27.3 %) and C. parapsilosis sensu lato (9.8 %). The overall resistance to fluconazole was 7.6 %, ranging from 3.9 % in C. albicans to 20.0 % in C. tropicalis. Only one C. glabrata isolate was resistant to the echinocandins. Four days after blood culture positivity, 99.7 % of the identifications and 90.3 % of the antifungal profiles were reported to the treating clinician. Candidaemia incidence rates differed up to 20-fold among Belgian hospitals; no clear factors explaining this difference were identified. The overall antifungal resistance rates were low but high azole resistance rates were recorded in C. tropicalis. [ABSTRACT FROM AUTHOR]

Objectives To assess the frequency and diversity of extended-spectrum β-lactamases (ESBLs) in Enterobacteriaceae isolates in Belgium. Methods During 2006 and 2008, non-duplicate clinical isolates of Enterobacteriaceae resistant to ceftazidime and/or cefotaxime were collected in 100 Belgian hospitals. ESBL production was confirmed by phenotypic and genotypic tests. MICs of 13 antimicrobial agents were determined by Etest. ESBL-encoding genes were identified by PCR sequencing and the blaCTX-M environment was characterized by PCR mapping. Selected isolates were genotyped by PFGE, multilocus sequence typing analysis and phylogenetic grouping by PCR. Results Overall, 733 isolates were confirmed as ESBL producers. Carbapenems and temocillin were active against ≥95% of all tested isolates. Co-resistance to co-trimoxazole and to ciprofloxacin was found in almost 70% and 80% of the strains, respectively. Overall, Escherichia coli (49%), Enterobacter aerogenes (32%) and Klebsiella pneumoniae (9%) represented the most prevalent species. Isolates harboured predominantly TEM-24 (30.7%), CTX-M-15 (24.2%) and TEM-52 (12.1%). Compared with 2006, the proportion of CTX-M-type enzymes increased significantly in 2008 (54% versus 23%; P < 10−6), mostly linked to a rising proportion of CTX-M-15-producing E. coli. TEM-24 decreased (19% in 2008 versus 43% in 2006; P < 10−6) during the same period, while the prevalence of TEM-52 remained unchanged (10% in 2008 versus 14% in 2006; not significant). Over 80% of the CTX-M-15-producing E. coli isolates clustered into a single PFGE type and phylogroup B2, corresponding to the sequence type (ST) 131 clone. Intra- and inter-species gene dissemination (CTX-M-15, CTX-M-2 and CTX-M-9) and wide epidemic spread of the CTX-M-15-producing E. coli ST131 clone in several Belgian hospitals were observed. Conclusions The rapid emergence of multiresistant CTX-M-15-producing E. coli isolates is of major concern and highlights the need for further surveillance in Belgium. [ABSTRACT FROM PUBLISHER]

Objectives: To investigate the presence of extended-spectrum β-lactamases (ESBLs) among Pseudomonas aeruginosa clinical isolates referred to two Belgian reference laboratories. Methods: Antibiograms were analysed for P. aeruginosa isolates referred between 2004 and 2008. Isolates resistant to ceftazidime (MIC.8 mg/L) and with a positive double-disc synergy test between ceftazidime and clavulanate were serotyped and screened for the presence of ESBL-encoding genes. Genes encoding metallo-b-lactamases (blaMBL) were sought by PCR in ESBL-producing isolates with positive imipenem/EDTA synergy tests. PFGE of SpeI-digested genomic DNA was used to compare isolates and selected strains were characterized by multilocus sequence typing. Results: Forty-eight (2.2%) of 2150 P. aeruginosa isolates were confirmed as class A ESBL-producing isolates by molecular testing. blaBEL and blaPER alleles were detected, respectively, in 39 and 10 P. aeruginosa isolates originating from 16 hospitals (two isolates were simultaneously positive for BEL and PER). Fifteen of the isolates were found to co-produce ESBLs and VIM carbapenemases. These strains were pan-resistant and remained susceptible only to colistin (MICs≤2 mg/L). The majority of the ESBL-producing isolates belonged to the same PFGE clone and were identified as ST235; serotype O11. Conclusions: BEL enzymes were produced by 80% of P. aeruginosa isolates with phenotypic evidence of ESBL production. BEL or PER ESBLs co-existed with VIM carbapenemases in 15 isolates and caused outbreaks in four hospitals. Our data further highlight the epidemic potential of the international clone ST235, which may have acquired blaBEL-1 gene cassettes from a yet unidentified local gene reservoir. [ABSTRACT FROM AUTHOR]

Moellerella wisconsensis, a member of the Enterobacteriaceae family, is rarely isolated in clinical specimens. We report here a case of M. wisconsensis infection in a 46-year-old cirrhotic patient with acute cholecystitis. This is the first reported case of a M. wisconsensis infection in Belgium and the first reported case of human bacteremia caused by this bacterium. Our case report is followed by a review of the literature. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented about three cases of Klebsiella pneumoniae carbapenemase (KPC)-2 producing sequence type (ST) 258 K. pneumoniae in Belgium.

Schools have been a point of attention during the pandemic, and their closure one of the mitigating measures taken. A better understanding of the dynamics of the transmission of SARS-CoV-2 in elementary education is essential to advise decisionmakers. We conducted an uncontrolled non-interventional prospective study in Belgian French-speaking schools to describe the role of attending asymptomatic children and school staff in the spread of COVID-19 and to estimate the transmission to others. Each participant from selected schools was tested for SARS-CoV-2 using a polymerase chain reaction (PCR) analysis on saliva sample, on a weekly basis, during six consecutive visits. In accordance with recommendations in force at the time, symptomatic individuals were excluded from school, but per the study protocol, being that participants were blinded to PCR results, asymptomatic participants were maintained at school. Among 11 selected schools, 932 pupils and 242 school staff were included between January and May 2021. Overall, 6449 saliva samples were collected, of which 44 came back positive. Most positive samples came from isolated cases. We observed that asymptomatic positive children remaining at school did not lead to increasing numbers of cases or clusters. However, we conducted our study during a period of low prevalence in Belgium. It would be interesting to conduct the same analysis during a high prevalence period.

In-center maintenance hemodialysis (HD) patients are at high risk of acquiring coronavirus disease 2019 (COVID-19) by cross-contamination inside the unit. The aim of this study was to assess retrospectively the dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission during the very first pandemic phase (March-July 2020) in a cohort of in-center maintenance HD patients and in nurses the same HD facility, using a phylogenetic approach. All SARS-CoV-2 quantitative reverse-transcription polymerase chain reaction positive patients and nurses from our HD unit-respectively 10 out of 98, and 8 out of 58- and two other positive patients dialyzed in our self-care unit were included. Whole-genome viral sequencing and phylogenetic analysis supported the cluster investigation. Five positive patients were usually dialyzed in the same room and same shift before their COVID-19 diagnosis was made. Viral sequencing performed on 4/5 patients' swabs showed no phylogenetic link between their viruses. The fifth patient (whose virus could not be sequenced) was dialyzed at the end of the dialysis room and was treated by a different nurse than the one in charge of the other patients. Three nurses shared the same virus detected in both self-care patients (one of them had been transferred to our in-center facility). The epidemiologically strongly suspected intra-unit cluster could be ruled out by viral genome sequencing. The infection control policy did not allow inter-patient contamination within the HD facility, in contrast to evidence of moderate dissemination within the nursing staff and in the satellite unit. Epidemiologic data without phylogenetic confirmation might mislead the interpretation of the dynamics of viral spreading within congregate settings., (© 2021 Wiley Periodicals LLC.)

This study aims to evaluate SARS-CoV-2 seroprevalence among health care workers (HCWs) and to assess self-reported risk factors for seropositivity. A total of 3255 HCWs were included and the overall seroprevalence was 7.8%. The likelihood of seropositivity was higher in participants reporting any COVID-19 symptoms within the last 4 months (OR 8.32, 95% CI 5.83-11.88, P < 0.001). Being a female HCW (OR 1.32, 95% CI 1.11-2.32, P < 0.01), having a cohabitant who was infected with SARS-CoV-2 (OR 2.55, 95% CI 1.78-3.66 P < 0.001) or a cohabitant who was a nursing home caregiver (OR 3.71, 95% CI 1.59-8.65, P = 0.002) were independently associated with an increased risk of seropositivity. Working in a COVID-19 unit (OR 1.64, 95% CI 1.21-2.23, P < 0.001) and being exposed to a SARS-CoV-2 infected co-worker (OR 1.30,95% CI 0.97-1.74, P = 0.016) resulted in higher seropositivity rate. Even if in-hospital exposure may play a significant role, increased infection risk is most likely attributable to household contact., Competing Interests: Declaration of competing interests None., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Purpose: Broth microdilution (BMD) stays as the reference testing method for determination of antimicrobial susceptibility testing (AST) to colistin and is considered essential for patient management and for monitoring of colistin resistance. This multicenter study aimed to evaluate the performance of automated systems for colistin AST among Enterobacterales as an alternative for BMD since the majority of laboratories use automated systems as first-line method., Methods: Twenty colistin resistant (COL-R) including 10 MCR producers and 10 colistin-susceptible (COL-S) Enterobacterales isolates were blindly tested for colistin susceptibility with the routine automated AST systems used by 8 laboratories (3 with BD Phoenix, 3 with Vitek2 and 2 with MicroScan). Additionally, 3 reference strains (E. coli ATCC 25922, E. coli NCTC 13846, and one COL-R mcr-negative K. pneumoniae M/14750) were tested in triplicate by each laboratory., Results and Conclusion: Results were compared with BMD performed at the reference laboratory. BD Phoenix and MicroScan automated AST systems provide accurate and reproducible categorical results for the testing of colistin in Enterobacterales. However, Vitek2 system showed poor performance for the detection of COL-R isolates especially those with MICs close to the susceptibility breakpoint (categorical agreement of 88% and precision categorical agreement of 81%).

Background: Healthcare workers (HCWs) are at high risk of acquiring COVID-19 and could play a role in nosocomial transmission. Since 4 th February 2020, Belgian Health authorities reported more than 90,568 cases, of which 8.3% were HCWs. Data on clinical characteristics, sources of infection and humoral immune response of HCWs with COVID-19 remain scarce., Aim: To analyse the clinical characteristics, humoral immune response, sources of contamination, and outcomes among HCWs with COVID-19., Methods: This retrospective study included 176 HCWs with laboratory-confirmed COVID-19 in a teaching hospital in Belgium. Between 1 st March and 31 st May 2020, all HCWs with symptoms suspected of COVID-19 were tested by reverse transcription polymerase chain reaction on a nasopharyngeal swab. Serological testing was performed between 55 and 137 days after the onset of symptoms., Findings: Median age was 40.8 years and 75% were female. Median delay between onset of symptoms and diagnosis was 4.39 days. Most frequent symptoms were cough and headache (both 75%). Fever accounted for 68.7%. Most represented professions were nurses (42%). HCWs were mainly infected by patient contact (32.9%); 7.6% required hospitalization and 1.7% were admitted to the intensive care unit. Unfortunately, one HCW died (0.5%). Total antibodies were positive in 109/126 (86.5%)., Conclusions: Clinical presentation of COVID-19 in HCWs does not differ from the general population. However, outcomes were more favourable with a mortality rate lower than that reported in Belgian COVID-19 patients in general (16%). The main source of infection was the hospital setting. Our positive antibodies rate was high but lower than previously reported., (Copyright © 2020 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

We evaluated the performance of the automated BD Phoenix CPO Detect test (BD-CPO test) for detection and Ambler classification of carbapenemases in Enterobacteriaceae, P.aeruginosa and A.baumannii complex. A collection of 287 Gram-negative clinical isolates, with a reduced susceptibility to at least one carbapenem including 184 carbapenemase-producing organisms (CPO) and 103 non-CPO, was tested. The BD-CPO test showed an overall sensitivity of 89.7% and specificity of 83.5% for carbapenemase detection. 1/7 of class A, 82.9% of class B, and 89.8% of class D carbapenemases were correctly classified. Poor detection sensitivity of 68.9% and specificity of 62.1% in P.aeruginosa was observed. However, combination with ceftazidime/avibactam susceptibility, provided by this panel, increased the performances for P.aeruginosa. The integration of an automated carbapenemase detection and classification in routine susceptibility panels would save time and help for therapeutic management. Further developments are needed to improve the accuracy of the BD-CPO test., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Mycobacterium abscessus is notorious for being intrinsically resistant to most antibiotics. Antibiotic efflux is one of the mechanisms used by M. abscessus to pump out antibiotics from their cells. Inhibiting efflux pumps (EPs) can be an attractive strategy to enhance the activity of drugs. The objective of this study is to determine the activity of EP inhibitors (EPIs) to enhance the efficacy of the new drug bedaquiline against M. abscessus clinical isolates., Methods: A total of 31 phenotypically and genotypically identified M. abscessus subsp. abscessus, M. abscesss subsp. massiliense, and M. abscessus subsp. bolletii clinical isolates were studied. The contribution of EPs was determined by investigating the minimum inhibitory concentration (MIC) levels of bedaquiline reduction in the absence and presence of EPIs verapamil and reserpine using the resazurin microtiter assay., Results: The observed bedaquiline MIC reduction by verapamil was observed in 100% isolates and by reserpine in 54.8% isolates. Bedaquiline MIC was 4-32-fold using verapamil with M. abscessus subsp. bolletii showing the highest fold change and between 2- and 4-fold using reserpine., Conclusions: The results obtained in this study confirm that bedaquiline MIC decreased in the presence of EPIs verapamil and reserpine in clinical isolates of M. abscessus. Verapamil was the most effective EPI. As shown in previous studies, verapamil may have clinical potential as adjunctive therapy to enhance the effect of bedaquiline., Competing Interests: None

Background: Nontuberculous mycobacteria (NTM) have been identified in human pulmonary and extrapulmonary infections and are increasing globally, which makes it challenging to identify them. This article reports our experience with the laboratory identification of NTM in clinical practice among pulmonary and extrapulmonary samples received in our routine work., Methods: The study was conducted at the Université Catholique de Louvain at the Cliniques Universitaires Saint-Luc, Brussels, Belgium, from 2015 to 2018. A total of 386 clinical samples were collected from patients suspected of having pulmonary or extrapulmonary mycobacterial infections. Routine laboratory methods phenotypic and molecular tests were performed., Results: The majority of NTM species were isolated from pulmonary samples (68%). The most prevalent species identified were Mycobacterium chimaera&#95;intracellulare group (32%), followed by Mycobacterium avium complex (21%), Mycobacterium abscessus complex (18%), Mycobacterium gordonae (9%), and Mycobacterium chelonae (4%). In extrapulmonary samples, M. avium and M. chimaera&#95;intracellulare were the most frequently isolated., Conclusion: The species diversity of NTM found in our setting suggests the importance of the use of new modern methods for accurate identification of NTM at species level and in some case at subspecies level for the proper treatment and management of patients., Competing Interests: None

Background: Rapid and effective screening of carbapenemase-producing Enterobacteriaceae (CPE) appears essential for adequate patient management and rapid implementation of infection control measures. Most of these screening techniques require a minimum of 24 h of culture. Molecular assays are an exception since these can be achieved within 1 h, but are expensive and usually require specialized facilities and trained personnel. In this context, lateral immunochromatography performed directly from rectal swab samples could represent a cost-effective alternative with a reduced turnaround time., Objectives: In this study, we assessed the performance of the OKN K-SeT test (Coris BioConcept, Gembloux, Belgium) for the rapid detection of OXA-48, KPC and NDM CPE directly from rectal swab samples., Methods: A total of 149 residual rectal swabs, routinely screened for CPE through selective culture and confirmed by PCR, were tested with a defined protocol consisting of a 2.5 h incubation of the swab in an enrichment medium containing meropenem followed by OKN K-SeT testing after centrifugation., Results: This method displayed an overall sensitivity of 96% and a specificity of 100% with a limit of detection ranging between 104 and 105 cfu/mL., Conclusions: Whereas this assay appears highly specific, it displays a reduced sensitivity compared with the standard procedure encompassing a culture step. Nonetheless, this rapid method allows an accelerated identification of most CPE carriers at a lower cost and, accordingly, the implementation of early appropriate management procedures.

Pseudomonas aeruginosa is a major cause of morbidity and mortality in cystic fibrosis patients. This study compared the antimicrobial susceptibilities of 153 P. aeruginosa isolates from the United Kingdom (UK) (n = 58), Belgium (n = 44), and Germany (n = 51) collected from 118 patients during routine visits over the period from 2006 to 2012. MICs were measured by broth microdilution. Genes encoding extended-spectrum β-lactamases (ESBL), metallo-β-lactamases, and carbapenemases were detected by PCR. Pulsed-field gel electrophoresis and multilocus sequence typing were performed on isolates resistant to ≥3 antibiotic classes among the penicillins/cephalosporins, carbapenems, fluoroquinolones, aminoglycosides, and polymyxins. Based on EUCAST/CLSI breakpoints, susceptibility rates were ≤30%/≤40% (penicillins, ceftazidime, amikacin, and ciprofloxacin), 44 to 48%/48 to 63% (carbapenems), 72%/72% (tobramycin), and 92%/78% (colistin) independent of patient age. Sixty percent of strains were multidrug resistant (MDR; European Centre for Disease Prevention and Control criteria). Genes encoding the most prevalent ESBL (BEL, PER, GES, VEB, CTX-M, TEM, SHV, and OXA), metallo-β-lactamases (VIM, IMP, and NDM), or carbapenemases (OXA-48 and KPC) were not detected. The Liverpool epidemic strain (LES) was prevalent in UK isolates only (75% of MDR isolates). Four MDR sequence type 958 (ST958) isolates were found to be spread over the three countries. The other MDR clones were evidenced in ≤3 isolates and localized in a single country. A new sequence type (ST2254) was discovered in one MDR isolate in Germany. Clonal and nonclonal isolates with different susceptibility profiles were found in 20 patients. Thus, resistance and MDR are highly prevalent in routine isolates from 3 countries, with meropenem, tobramycin, and colistin remaining the most active drugs., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Objectives: Typhoid fever (TF) occurs rarely in industrialized countries because of advances in health care and improvement of drinking water. Conversely, non-typhoid salmonellosis (NTS) remains widespread, because of food contamination or asymptomatic carriage. Non-typhoid salmonellosis can be severe when becoming invasive non-typhoid salmonellosis (iNTS). Although established prognostic indicators of the two pathologies are different, direct comparisons between iNTS and TF in the literature remain scarce. The purpose of this study was to analyse and compare demographic, clinical features and outcome of hospitalized patients with TF and iNTS., Methods: Demographic, clinical features and outcome were retrospectively analysed in a series of patients hospitalized between 2007 and 2012., Results: A total of 33 patients were enrolled, including 13 with established TF and 20 with iNTS. No differences between TF and iNTS patients were observed in incidence of fever, abdominal cramps, diarrhoea, headache, nausea and vomiting and duration of antibiotic therapy (≤ 7 days in both groups). Although the clinical outcome of TF patients was identical to that of iNTS patients, including incidence of complications, length of hospitalization and mortality (1/13 (7.7%) vs 2/20 (10%), P = 0.43), comorbidities were less frequent in the TF group than in the iNTS group (2/13 vs 15/20, P = 0.003)., Conclusion: The clinical profile and outcome of TF patients are similar to those with iNTS. Although comorbidities are more often associated with iNTS, the results of our study suggest that clinical management of these two diseases should remain similar.

Infections caused by multi drug resistant bacteria (MDRB) constitutes an international health care problem. Since the year 2000, a longitudinal surveillance programme (LSP) and two multicentric surveys (100 hospitals, 826 isolates) were performed to monitor the emergence of MDRB in Belgium. The implementation of a LSP detected the emergence and spread of new types of ESBLs (CTX-M), mostly among community associated E. coli in the setting of a university hospital several years before the large spread and recognition in Belgium of a pathogenic E. coli CTX-M-15 (B2-O25:H4-ST131) pandemic clone (found in extra-intestinal virulent strains). This finding supports the progressive increase in Belgium of systemic infections including UTI caused by MDRB with limited therapeutical options. The real burden of the problem remains however, difficult to estimate in the absence of any surveillance network in Belgium to monitor the epidemiology of antimicrobial resistance in the community. The current Belgian national recommendations for the detection, surveillance, prevention and control of epidemics by ESBL-producing organisms and possibly other MDRBs (eg: Carbapenemase producing Enterobacteriaceae [CPE]) must be updated taking into accounts these new elements. A global coordinated network for antimicrobial surveillance resistance gathering experts (e.g: public health epidemiologists, representative of the national reference centres of antimicrobial resistance, field experts in infection control, infectious disease specialists, other clinicians and general practitioners) must be urgently implemented, including the longitudinal analysis of resistance in different ecosystems (human, animal, water and food).

Background: Septic arthritis (SA) is a rheumatological emergency that can lead to rapid joint destruction and irreversible loss of function. The most common pathogen causing SA is Staphylococcus aureus which is responsible for 37-65% of cases. Streptococcus pneumoniae is traditionally described as an uncommon cause of SA of a native joint. The objective of our study was to analyse clinical characteristics, treatment, and outcome of all cases of pneumococcal septic arthritis treated in our institution, and to compare them with other series published in the literature., Materials and Methods: We conducted a retrospective study of pneumococcal SA identified among all cases of SA diagnosed in a teaching hospital of one thousand beds between 2004 and 2009. Diagnosis was based on culture of joint liquid or by the presence of pneumococcal bacteraemia and purulent (more than 50 000/mm(3) white blood cells with more than 90% neutrophils) joint fluid aspiration., Results: Among 266 cases of SA, nine patients (3·3%) were diagnosed as having pneumococcal SA. The median age was 75 years. The main affected joint was the knee (7/9). No patient had more than one joint involved. Four patients suffered from concomitant pneumonia. Joint culture and blood cultures were positive in 7/9 and 5/9, respectively. Median (range) length of stay was 18 days (3-47 days). One patient with associated pneumococcal bacteraemia died 19 days after admission. Seven patients recovered completely., Conclusions: Streptococcus pneumoniae is now being increasingly recognized as a common agent of SA. This organism is frequently associated with pneumococcal pneumonia or bacteraemia, particularly in patients with advanced age and comorbidities. Direct inoculation of joint fluid into blood culture medium BACTEC system increases the probability of microbiological diagnosis. The prognosis is usually favourable if the disease is promptly recognized and treated (antibiotic therapy combined with joint drainage).

Objectives: To collect recent data on the susceptibility of anaerobes to antimicrobial agents with known activity against anaerobes, and to compare them with results from previous Belgian multicentre studies., Methods: Four hundred and three strict anaerobic clinical isolates were prospectively collected from February 2011 to April 2012 in eight Belgian university hospitals. MICs were determined by one central laboratory for 11 antimicrobial agents using Etest methodology., Results: According to EUCAST breakpoints, >90% of isolates were susceptible to amoxicillin/clavulanate (94%), piperacillin/tazobactam (91%), meropenem (96%), metronidazole (92%) and chloramphenicol (98%), but only 70% and 40% to clindamycin and penicillin, respectively. At CLSI recommended breakpoints, only 71% were susceptible to moxifloxacin and 79% to cefoxitin. MIC50/MIC90 values for linezolid and for tigecycline were 1/4 and 0.5/4 mg/L, respectively. When compared with survey data from 2004, no major differences in susceptibility profiles were noticed. However, the susceptibility of Prevotella spp. and other Gram-negative bacilli to clindamycin decreased from 91% in 1993-94 and 82% in 2004 to 69% in this survey. Furthermore, the susceptibility of clostridia to moxifloxacin decreased from 88% in 2004 to 66% in 2011-12 and that of fusobacteria from 90% to 71%., Conclusions: Compared with previous surveys, little evolution was seen in susceptibility, except a decline in activity of clindamycin against Prevotella spp. and other Gram-negative bacteria, and of moxifloxacin against clostridia. Since resistance was detected to all antibiotics, susceptibility testing of anaerobic isolates is indicated in severe infections to confirm appropriateness of antimicrobial therapy.

Capnocytophaga species are part of the normal human oral bacterial flora.They are recognized as opportunistic pathogens leading to various extra-oral infections including septicemia, osteomyelitis, abscesses and keratitis and they have been rarely reported as a cause of chorioamionitis and neonatal infection. We here report the first two cases of chorioamionitis produced by Capnocytophaga sputigena and the recently described C. leadbetteri in Belgium. Both isolates were correctly identified at the genus level, in the first 24 hours of incubation by MALDI-TOF.

Objective: To characterize the genetic determinants associated with an AmpC phenotype in clinical Escherichia coli isolates., Methods: E. coli strains recovered at two Belgian hospitals between 2004 and 2006 were selected on the basis of an AmpC-producing phenotype. Plasmid-mediated cephalosporinases coding genes and the sequence of chromosomal ampC genes were identified by PCR/sequencing. The isolates were submitted to phylotyping and genotyping analysis using rep-PCR (Diversilab) and PFGE. A novel chromosomal ampC gene was cloned., Results: Eighty-three out of 6850 E. coli isolates were selected. Seventy-two isolates were found to overexpress their chromosomal cephalosporinases while 11 contained plasmid-mediated cephalosporinases. Among chromosomal AmpC overproducers, 12 were extended-spectrum AmpC (ESAC) expressing isolates which all displayed reduced susceptibility to cefepime. Cloning of a new ESAC allele suggested that L293P mutation was responsible of the extension of the hydrolysis spectrum to cefepime. AmpC overproducers, including ESAC producers, predominantly belonged to phylogenetic group A and B1, while plasmid-mediated AmpC-producing isolates preferentially belong to phylogroup B2 and D. According to rep-PCR, the majority of the E. coli isolates belonging to phylogroup A were clonally related which was further confirmed by PFGE for the 11 ESAC expressing isolates., Conclusions: Chromosomal AmpC overproduction was the most common resistance mechanism, and the occurrence of ESAC was found to be as frequent as plasmid-mediated cephalosporinases. The detection of a new ESAC allele, of an ESAC producing strain belonging to phylogroup D and the existence of a clonal relationship between ESAC producing strains underline the need for study of the clinical relevance of this mechanism of resistance., (Copyright (c) 2009 Elsevier Masson SAS. All rights reserved.)

Objective: To describe an outbreak of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae in the intensive care units (ICUs) of a hospital and the impact of routine and reinforced infection control measures on interrupting nosocomial transmission., Design: Outbreak report., Setting: A 31-bed intensive care department (composed of 4 ICUs) in a university hospital in Belgium., Intervention: After routine infection control measures (based on biweekly surveillance cultures and contact precautions) failed to interrupt a 2-month outbreak of ESBL-producing K. pneumoniae, reinforced infection control measures were implemented. The frequency of surveillance cultures was increased to daily sampling. Colonized patients were moved to a dedicated 6-bed ICU, where they received cohorted care with the support of additional nurses. Two beds were closed to new admissions in the intensive care department. Meetings between the ICU and infection control teams were held every day. Postdischarge disinfection of rooms was enforced. Broad-spectrum antibiotic use was discouraged., Results: Compared with a baseline rate of 0.44 cases per 1,000 patient-days for nosocomial transmission, the incidence peaked at 11.57 cases per 1,000 patient-days (October and November 2005; rate ratio for peak vs baseline, 25.46). The outbreak involved 30 patients, of whom 9 developed an infection. Bacterial genotyping disclosed that the outbreak was polyclonal, with 1 predominant genotype. Reinforced infection control measures lasted for 50 days. After the implementation of these measures, the incidence fell to 0.08 cases per 1,000 patient-days (rate ratio for after the outbreak vs during the outbreak, 0.11)., Conclusion: These data indicate that, in an intensive care department in which routine screening and contact precautions failed to prevent and interrupt an outbreak of ESBL-producing K. pneumoniae, reinforced infection control measures controlled the outbreak without major disruption of medical care.

Temocillin is a narrow spectrum penicillin with high stability to most beta-lactamases including AmpC types and extended-spectrum types (ESBLs). We have analysed its in vitro activity against 652 clinical isolates of Enterobacteriaceae prospectively collected from patients hospitalised in intensive care units at seven different university hospitals in Belgium in 2005. Strains were screened for ESBL production using cefotaxime and ceftazidime screen agar plates and by double ESBL E-tests. The MIC of temocillin and of five comparators was determined using the E-test method. ESBLs were characterized at one central laboratory by isoelectric focusing, PCR for bla genes of the SHV, TEM, and CTX-M families, and by DNA sequencing. The prevalence of ESBL-producing Enterobacteriaceae averaged 11.8% and ranged between 3.0 and 29% in the different hospitals. Meropenem exhibited the highest in vitro activity overall (mode MIC 0.064 microg; MIC(90); 0.19 microg/ml), whereas ceftazidime (MIC(90) > 256 microg/ml) and ciprofloxacin (MIC(90) > 32 microg/ml) scored the worst. Temocillin was active against more than 90% of the isolates including most AmpC- and ESBL-producing isolates. These data indicate the well preserved activity of temocillin over the years against Enterobacteriaceae and show the wide variation in prevalence of ESBL-producing Enterobacteriaceae isolates in Belgian intensive care units. Prospective clinical studies are, however, needed to validate the usefulness of temocillin in the treatment of microbiologically documented infections caused by ESBL- and/or AmpC- overproducing nosocomial Enterobacteriaceae pathogens.

Bacterial resistance to antibiotics is as old as the living planet, i.e. over 3 billion years ago when bacterial life emerged together with its biological warfare weapons, the antibiotics. Bacteria can readily develop a diverse array of antibiotic resistance mechanisms, including selection of mutants which produce modified low-affinity drug targets, exhibit reduced drug permeability or increased drug efflux. Another pathway is through acquisition of mobile genetic determinants of drug inactivating enzymes, target protection or bypass mechanisms. Antibiotic resistance has accelerated and spread across the community of human and animal bacteria following massive use of antibiotics for over half-a century in human and veterinary medicine and agriculture. Today, the increasing prevalence of resistance among many bacterial pathogens is no longer counter-balanced by the launching of innovative and active antibiotics. This imbalanced "arms race" undermines the escalating use of broad-spectrum antibiotics and increases the risk of therapeutic failure, drug side effets and excess costs of care. The accumulation of antibiotic resistance genes and their epidemic dissemination is fuelled by the selective pressure associated with excess and inappropriate use of antibiotics combined with insufficient infection prevention and control measures. Confronting this threat effectively requires new strategies to be implemented globally, nationally, and locally across all healthcare and agricultural sectors.

Objectives: Temocillin is a semi-synthetic 6-alpha-methoxy derivative of ticarcillin. It is highly stable to most bacterial beta-lactamases. However, data concerning its activity against extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae are limited. We have analysed the in vitro activity of temocillin against clinical isolates of ESBL-producing Escherichia coli over the past 4 years in a university hospital in Brussels, Belgium., Methods: Strains were screened for ESBL production using the double-disc synergy test. The MIC of 12 antimicrobial agents was determined for ESBL-producing E. coli isolates (n = 162) using the agar dilution method. ESBLs were characterized by isoelectric focusing; multiplex PCR for bla genes of the SHV, TEM and CTX-M families; and DNA sequencing., Results: ESBL-producing E. coli isolates harboured CTX-M+TEM (35%), TEM alone (44%), CTX-M alone (6%), CTX-M+SHV (2%) and other ESBL combinations (10%). The proportion of temocillin-susceptible isolates was 92%, with MIC(50) and MIC(90) values of 8 and 32 mg/L, respectively. Co-resistance to ciprofloxacin and co-trimoxazole in ESBL-producing E. coli was frequent (39%). The proportion of isolates not susceptible to aminoglycosides was 55, 37 and 4% for tobramycin, gentamicin and amikacin, respectively. The proportion of isolates not susceptible to ceftazidime, cefotaxime, cefepime and piperacillin/tazobactam was 70, 52, 37 and 11%, respectively. No resistance to meropenem was observed. The proportion of strains exhibiting resistance to temocillin by year was stable over the study period., Conclusions: These data indicate good in vitro activity of temocillin against multiresistant ESBL-producing E. coli. Prospective clinical studies are necessary to examine temocillin's potential role in the treatment of non-complicated infections caused by ESBL-producing E. coli.

From the outset, the Department of Microbiology of Erasme Hospital has striven to offer to clinical teams a full range of high performance tests for diagnosis and therapeutic monitoring of bacterial, viral, fungal and parasitic infections, including opportunistic infections. Furthermore, the laboratory has developed and made available cutting edge tools for the epidemiologic surveillance and outbreak investigation in support to the hospital infection control and antibiotic resistance control programmes. Research and development programmes have led to technological innovation in the fields of molecular virologic diagnosis, notably for HIV and herpes viruses, rapid bacterial identification, detection of resistance genes and epidemiological typing. Research in collaboration with clinical teams has focused on congenital infections, HIV infection, opportunistic infections and prevention of nosocomial infection. The Department acts as national reference laboratory for a number of infectious diseases and co-ordinates or actively participates to several European multicentre studies and epidemiologic surveillance networks.