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With the increase in antimicrobial resistance, fast reporting of antimicrobial susceptibility testing (AST) results is becoming increasingly important. EUCAST developed a method for rapid AST (RAST) directly from the broth of positive blood cultures (BC). Inhibition zones are read after 4, 6, and 8 h, with specific breakpoints per time point. We evaluated the RAST method based on EUCAST disk diffusion methodology with inoculation of BC broth using WASPLab® (inclusive Colibrí™ and Radian®). Forty-nine non-duplicate strains were tested: Escherichia coli n = 17, Klebsiella pneumoniae n = 7, Pseudomonas aeruginosa n = 4, Acinetobacter baumannii n = 2, Staphylococcus aureus n = 10, Enterococcus faecalis n = 6, and Enterococcus faecium n = 3. Results were compared to direct AST and standardized AST. Good categorical agreement was obtained at all time points for all groups, except P. aeruginosa. RAST cut-offs for extended-spectrum β-lactamase (ESBL)-producing Enterobacterales enabled the detection of all included ESBL isolates (n = 5) at all time points, except for 1 E. coli ESBL after 4 h. RAST cut-offs for carbapenemase-producing Enterobacterales enabled the detection of only one carbapenemase after 6 h. However, all carbapenemases (n = 3) were correctly detected after 8 h. Two methicillin-resistant S. aureus were included; both were correctly categorized as cefoxitin-resistant at 6 and 8 h. At 4 h, there was insufficient growth for inhibition zone interpretation. EUCAST RAST is a fast supplementary tool for direct AST of positive BC. WASPLab® provides a significant advantage as pictures are made automatically implicating that we are not strictly bound to the time points for inhibition zone interpretation.

Recently, Copan (Italy) introduced the Colibrí instrument for automated colony picking and preparation of matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) target plates. Our study aimed to validate this system for yeasts as such testing has not been performed yet and is a missing link needed to implement the system for routine use. Fifty-five Candida strains were selected to evaluate the accuracy of Colibrí. For each strain, a sheep blood agar plate supplemented with X and V factors (HEM) and a Sabouraud agar plate (SAB) were inoculated and incubated using the WASPlab specimen processing system (Copan). After 18 h and 36 h of incubation, the isolates were spotted in parallel using Colibrí and manually onto MALDI-TOF target plates with the addition of formic acid and identified using MALDI-TOF mass spectrometry. The reproducibility was evaluated using ATCC reference and clinical isolate-derived strains. The cumulative percentage of acceptable identification scores (IDs) after 36 h was 91%for strains cultured on HEM plates using both Colibrí and the manual method. The SAB plates showed inferior results for both Colibrí (76%) and the manual method (78%). We observed an overall agreement of 92% at 18 h for identification of the strains on the HEM plates between Colibrí and the manual method and 94% after 36 h. For the SAB plates, the agreement was 78% after 18 h and 84% after 36 h. Apart from Candida dubliniensis and Candida tropicalis, all Candida species were identified with 100% accuracy using Colibrí on HEM plates. We observed good agreement between Colibrí and the manual reference method. These results demonstrate that Colibrí is a reliable system for MALDI-TOF target preparation for yeast identification, allowing increased standardization and less hands-on time.

BackgroundWorldwide, healthcare-associated SARS-CoV-2 infections are a major problem: they are associated with increased morbidity, mortality, and hospitalization costs. In-depth studies across the pandemic are crucial to understand and prevent transmission in hospital settings. The principal aims of this study were to characterise patients and validate ECDC definitions of healthcare-associated COVID-19 infections.MethodsWe set up a retrospective observational study spanning the first three waves of the COVID-19 pandemic in a Belgian university hospital: it describes the characteristics of COVID-19 patients admitted, with either healthcare- or community-associated infections. We performed a cluster analysis through epidemiological and viral genome analyses of the healthcare-associated infections, in order to validate the ECDC definitions of healthcare-associated COVID-19 infections.ResultsBetween week 10 of 2020 and week 22 of 2021, 168 patients were hospitalized with healthcare-associated COVID-19. The following factors were found more often in symptomatic healthcare- than in community-associated hospitalized patients: older age, increased frailty, smoking habits, and comorbidities. The genome-based cluster analyses showed that different viral lineages predominated in different timeframes. We observed a good correlation of epidemiological data with genome sequencing results in at least 12 different outbreaks in our hospital, thus validating the ECDC definitions. ConclusionsThis in-depth characterization sheds new light on the problem of healthcare-associated COVID-19 infections, in particular on patients’ characteristics, epidemiology, and cluster dynamics. Even though epidemiological evaluation of nosocomial infections is vital, management of nosocomial outbreaks can undoubtedly benefit from genome sequencing analyses to reinforce their strategy.

Background Hydroxychloroquine and chloroquine have been used for hospitalized coronavirus disease 2019 patients because of their antiviral and anti-inflammatory function. However, little research has been published on the impact of the immunomodulatory effect of (hydroxy)chloroquine on humoral immunity. Case presentation We report a case of symptomatic severe acute respiratory syndrome coronavirus 2 reinfection, diagnosed 141 days after the first episode, in a 56-year-old man of Black African origin treated with hydroxychloroquine for lupus erythematosus. No anti-severe acute respiratory syndrome coronavirus 2 IgG antibodies could be detected 127 days after the initial episode of coronavirus disease 2019. Conclusions The treatment with hydroxychloroquine probably explains the decreased immune response with negative serology and subsequent reinfection in our patient. As humoral immunity is crucial to fight a severe acute respiratory syndrome coronavirus 2 infection, the use of (hydroxy)chloroquine is likely to have a detrimental effect on the spread of the virus. This case emphasizes that more needs to be learned about the role of antibodies in protecting against severe acute respiratory syndrome coronavirus 2 (re)infection and the role of (hydroxy)chloroquine on humoral immunity.

In Western Europe, the incidence of both respiratory and cutaneous diphtheria, caused by toxin-producing Corynebacterium diphtheriae, Corynebacterium ulcerans or Corynebacterium pseudotuberculosis, has been low over the past few decades thanks to the use of an effective vaccine and a high level of vaccination coverage. However, the disease has still not been eradicated and continues to occur in all of Europe. In order to prevent sequelae or a fatal outcome, diphtheria antitoxin (DAT) should be administered to suspected diphtheria patients as soon as possible, but economic factors and issues concerning regulations have led to poor availability of DAT in many countries. The European Centre for Disease Prevention and Control and World Health Organization have called for European Union-wide solutions to this DAT-shortage. In order to illustrate the importance of these efforts and underline the need for continued diphtheria surveillance, we present data on all registered cases of toxigenic and non-toxigenic C. diphtheriae, C. ulcerans and C. pseudotuberculosis in Belgium during the past decade, up to and including 2017.

Background/aimsSARS-CoV-2 is highly contagious. More evidence concerning extrapulmonary transmission routes such as the eyes is urgently needed. Although the humoral immune response is important in the viral containment, the local response in tears has not yet been studied. The aim of our study was twofold: to assess the prevalence of both SARS-CoV-2 RNA and antibodies in tear fluid.MethodsIn a first series, nasopharyngeal sampling and tear sampling by Schirmer test strips were performed in 26 acutely ill patients with COVID-19 to assess the presence of SARS-CoV-2 RNA by reverse transcription PCR. In a second series, IgG and IgA responses to SARS-CoV-2 spike protein in serum and tear fluid of convalescent individuals (n=22) were compared with control individuals (n=15) by ELISA.ResultsSARS-CoV-2 RNA was detected in tears of 7/26 (26.9%) patients with COVID-19. None of them had ocular symptoms. Convalescent individuals displayed a significant higher ratio of IgG (pConclusionsOur results demonstrate the presence of SARS-CoV-2 RNA and a local IgG and IgA immune response in tear fluid. These data confirm the possibility of SARS-CoV-2 transmission through tear fluid and the importance of the eye as a first defence against SARS-CoV-2, indicating the potential of tears as a non-invasive surrogate for serum in monitoring the host immune response.

Penicillins, can be used in treatment of infections due to Prevotella species if they are susceptible to penicillin. Early and accurate preliminary detection of β-lactamase-producing isolates is crucial for treatment of infection. The aim of this study was to determine β-lactamase-producing Prevotella species by MALDI-TOF MS and screen them for the presence of cfxA gene, responsible for β-lactamase production. A total of 500 clinically relevant Prevotella isolates, collected from 13 countries for the previous European antibiotic resistance surveillance study, were tested. Susceptibility testing was performed against ampicillin and ampicillin/sulbactam by Etest methodology. EUCAST guidelines were used for susceptibility interpretations; the isolates with MIC value ≤ 0.5 for ampicillin were considered susceptible and >2 resistant. All Prevotella isolates, were tested for detection of β-lactamase activity by MALDI-TOF MS (Vitek® MS Research Use Only) system and the presence of the cfxA gene by PCR method. The susceptibility levels of the isolates to ampicillin/sulbactam and ampicillin were 99.6% and 43.4%, respectively. A total 59% of isolates presented β-lactamase activity and 60.8% were cfxA gene positive. Both these tests were positive for isolates in the resistant category. Additionally, >95% of the isolates (n = 65) which ampicillin MIC values ranged from >0.5 μg/mL to 2 μg/ml displayed β-lactamase activity. We also found that the MALDI-TOF MS-based β-lactamase assay delivers results in 2 h. We found a high concordance between the MALDI-TOF MS β-lactamase results in terms of cfxA β-lactamase gene presence. MALDI-TOF MS may serve as a simple and efficient alternative method of the existing phenotypic and PCR-based methods.

Summary Background Serratia marcescens is notorious for its increasing antimicrobial resistance and potential to cause outbreaks in neonatal intensive care units (NICUs). A promising tool in outbreak investigations is whole-genome sequencing (WGS). Objectives To describe a S. marcescens outbreak (2018–2019) in an NICU and discuss which infection control measures contributed to containment, addressing the potential of WGS. Methods S. marcescens isolates from patients and the environment isolated during the 2018–2019 NICU outbreak were analysed. In comparison, isolates from previous presumed NICU outbreaks and adult blood cultures were included. WGS and whole-genome multi-locus sequence typing analysis were performed. Results Sixty-three S. marcescens isolates were analysed. The 2018–2019 outbreak was divided into three clusters, including four environmental strains (drains, N=3; baby scale, N=1). The strains differed significantly from those of an NICU outbreak in 2014 and adult blood cultures. Besides standard infection control measures, the siphons were replaced and weekly decontamination was performed with acetic acid 10%. Seven acquired-resistance genes and 29 virulence-associated genes were detected. Conclusions It was assumed that both neonates and drains were reservoirs of S. marcescens cross-contamination via the hands of healthcare workers and parents. Initially, standard measures, including hand hygiene, were reinforced. However, definitive containment was achieved only after replacement of the siphons and weekly decontamination with acetic acid. WGS enables faster recognition of an outbreak with accurate mapping of the spread, facilitating the implementation of infection control measures. WGS also provides interesting information about the spread of antibiotic resistance and virulence genes.

A cluster of Legionnaires' disease (LD) with 10 confirmed, three probable and four possible cases occurred in August and September 2016 in Dendermonde, Belgium. The incidence in the district was 7 cases/100 000 population, exceeding the maximum annual incidence in the previous 5 years of 1.5/100 000. Epidemiological, environmental and geographical investigations identified a cooling tower (CT) as the most likely source. The case risk around the tower decreased with increasing distance and was highest within 5 km.Legionella pneumophilaserogroup 1, ST48, was identified in a human respiratory sample but could not be matched with the environmental results. Public health authorities imposed measures to control the contamination of the CT and organised follow-up sampling. We identified obstacles encountered during the cluster investigation and formulated recommendations for improved LD cluster management, including faster coordination of teams through the outbreak control team, improved communication about clinical and environmental sample analysis, more detailed documentation of potential exposures obtained through the case questionnaire and earlier use of a geographical information tool to compare potential sources and for hypothesis generation.

Here, we sought to assess the levels of antibiotic resistance among intestinal Bacteroides and Parabacteroides strains collected between 2014 and 2016 in Europe and also attempted to compare resistance levels between clinical and commensal isolates. Bacteroides and Parabacteroides isolates were recovered from faecal samples via the novel Bacteroides Chromogenic Agar (BCA) method. Antibiotic susceptibilities were determined by agar dilution for ten antibiotics. The values obtained were then statistically evaluated. Altogether 202 Bacteroides/Parabacteroides isolates (of which 24, 11.9%, were B. fragilis) were isolated from the faecal specimens of individuals taken from five European countries. The percentage values of isolates resistant to ampicillin, amoxicillin/clavulanate, cefoxitin, imipenem, clindamycin, moxifloxacin, metronidazole, tetracycline, tigecycline and chloramphenicol were 96.6, 4.5, 14.9, 2.0, 47.3, 11.4, 0, 66.2, 1.5 and 0%, respectively. These values are close to those reported in the previous European clinical Bacteroides antibiotic susceptibility survey except for amoxicillin/clavulanate and clindamycin, where the former was lower and the latter was higher in normal microbiota isolates. To account for these latter findings and to assess temporal effects we compared the data specific for Hungary for the same period (2014-2016), and we found differences in the resistance rates for cefoxitin, moxifloxacin and tetracycline. (C) 2020 Elsevier Ltd. All rights reserved.

We tested the in vitro susceptibility of ceftazidime-avibactam and ceftolozane-tazobactam and 13 other antibiotics against 91 Burkholderia cepacia complex (BCC) strains isolated from cystic fibrosis patients since 2012. The highest susceptibility (82%) was found for trimethoprim-sulfamethoxazole. Eighty-one and 63% of all BCC strains were susceptible to ceftazidime-avibactam and ceftolozane-tazobactam, respectively. For temocillin, ceftazidime, piperacillin-tazobactam, and meropenem, at least 50% of the strains were susceptible. B. stabilis seems to be more resistant than other BCC species.

Anaerobic infections are difficult to diagnose and treat, because of the often slow in vitro growth, the polymicrobial nature and the increasing antimicrobial resistance. Furthermore because of their fastidiousness, anaerobic bacteria often stay unrecognized in clinical practice. Clinical specimens potentially harboring these species require special handling to permit satisfactory recovery of these potential important pathogens. In a clinical setting, temporary storage and transportation to the laboratory are unavoidable before these specimens can be cultured. In the current study we expand the knowledge about the recovery of a wide range of clinically relevant anaerobic bacteria from an eSwab® container after different storage durations and temperatures. Our findings support the use of the eSwab® container as a relative short-term storage unit for anaerobic species. When stored at 2-4°C immediately after inoculation, all anaerobic species (except for Clostridium clostridioforme) can be recovered from the liquid Amies medium until 1day post-specimen collection. Because most samples in the clinical setting are processed in this time span, the eSwab® container is sufficiently capable of retaining viability in daily routine. However; because of inevitable centralization of clinical laboratories, adequate storage of these specimens for an extended period of time will be essential in the future. Therefore in certain cases, when viability is desired for longer periods (>1day), storage of the containers at 2-4°C is certainly advisable. KEYWORDS: Fastidious anaerobes; Viability recovery; eSwab PMID: 29146304 DOI: 10.1016/j.mimet.2017.11.009, Anaerobic infections are difficult to diagnose and treat, because of the often slow in vitro growth, the polymicrobial nature and the increasing antimicrobial resistance. Furthermore because of their fastidiousness, anaerobic bacteria often stay unrecognized in clinical practice. Clinical specimens potentially harboring these species require special handling to permit satisfactory recovery of these potential important pathogens. In a clinical setting, temporary storage and transportation to the laboratory are unavoidable before these specimens can be cultured. In the current study we expand the knowledge about the recovery of a wide range of clinically relevant anaerobic bacteria from an eSwab® container after different storage durations and temperatures. Our findings support the use of the eSwab® container as a relative short-term storage unit for anaerobic species. When stored at 2-4°C immediately after inoculation, all anaerobic species (except for Clostridium clostridioforme) can be recovered from the liquid Amies medium until 1day post-specimen collection. Because most samples in the clinical setting are processed in this time span, the eSwab® container is sufficiently capable of retaining viability in daily routine. However; because of inevitable centralization of clinical laboratories, adequate storage of these specimens for an extended period of time will be essential in the future. Therefore in certain cases, when viability is desired for longer periods (>1day), storage of the containers at 2-4°C is certainly advisable.

Fusobacterium nucleatum is anaerobic oral microbiota that might be associated with cancer. We reported 22 consecutive cases of patients (mean age of 63.8 years (range 34–89), 59.1% male) with F . nucleatum bacteremia that were admitted to a university hospital over a 10-year period. In 17 (77.2%) of these patients, F . nucleatum was the sole possible pathogen. Seven of the 22 patients (31.8%) had active cancer: esophagus carcinoma (n = 3), hematologic malignancies (n = 1), gastrointestinal stromal tumor (n = 1), melanoma (n = 1), and breast cancer (n = 1). In six out of seven patients (85.7%), the F . nucleatum was found within six months of the diagnosis of cancer. Four of seven (57.1%), patients with cancer were on chemotherapy. Three of 22 patients (13.4%) died within 1 month of F . nucleatum bacteremia due to cancer. In conclusion, F . nucleatum bacteremia occurs rarely and when it is found, it is often in patients with cancer, especially those with a recent diagnosis.

Background Carbapenemase-producing Enterobacteriaceae (CPE) are emerging pathogens that represent a major public health threat. In the University Hospital of Brussels, the incidence of new patients with CPE rose from 1 case in 2010 to 35 cases in 2015. Between January and August 2015, five patients became infected/colonized with CPE during their stay in the same room in the intensive care unit (ICU). Since the time period between those patients was relatively short and the strains belonged to different species with different antibiograms and mechanisms of resistance, the hypothesis was that the environment could be a possible source of transmission. Methods and results Environmental samples suggested that a contaminated sink was the source of the outbreak. Besides other strains, Citrobacter freundii type OXA-48 was frequently isolated from patients and sinks. To investigate the phylogenetic relationschip between those strains, pulsed-field gel electrophoresis was performed. The strains isolated from patients and the sink in the implicated room were highly related and pointed to sink-to-patient transmission. In total, 7 of 8 sinks in the isolation rooms of the ICU were found to be CPE contaminated. To control the outbreak, the sinks and their plumbings were replaced by new ones with another structure, they were flushed every morning with a glucoprotamin solution and routines regarding sink practices were improved leading to discontinuation of the outbreak. Conclusions This outbreak highlights that hospital sink drains can accumulate strains with resistance genes and become a potential source of CPE.

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.

The performance of matrix-assisted laser desorption–ionization time of flight mass spectrometry (MALDI-TOF MS) for species identification of Prevotella was evaluated and compared with 16S rRNA gene sequencing. Using a Bruker database, 62.7% of the 102 clinical isolates were identified to the species level and 73.5% to the genus level. Extension of the commercial database improved these figures to, respectively, 83.3% and 89.2%. MALDI-TOF MS identification of Prevotella is reliable but needs a more extensive database.

In order to identify current trends in anaerobic bacteraemia, a 10-year retrospective study was performed in the University Hospital Brussel, Belgium. All clinically relevant bacteraemia detected from 2004 until 2013 were included. Medical records were reviewed in an attempt to define clinical parameters that might be associated with the occurrence of anaerobic bacteraemia. 437 of the isolated organisms causing anaerobic bacteraemia were thawed, subcultured and reanalyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). There were an average of 33 cases of anaerobic bacteraemia per year during 2004–2008 compared to an average of 27 cases per year during 2009–2013 (P = 0.017), corresponding to a decrease by 19% between the first and the latter period. Also, the total number of cases of anaerobic bacteraemia per 100,000 patient days decreased from 17.3 in the period from 2004 to 2008 to 13.7 in the period 2009 to 2013 (P = 0.023). Additionally, the mean incidence of anaerobic bacteraemia decreased during the study period (1.27/1000 patients in 2004 vs. 0.94/1000 patients in 2013; P = 0.008). In contrast, the proportion of isolated anaerobic bacteraemia compared to the number of all bacteraemia remained stable at 5%. Bacteroides spp. and Parabacteroides spp. accounted for 47.1% of the anaerobes, followed by 14.4% Clostridium spp., 12.6% non-spore-forming Gram-positive rods, 10.5% anaerobic cocci, 8.2% Prevotella spp. and other Gram-negative rods and 7.1% Fusobacterium spp. The lower gastrointestinal tract (47%) and wound infections (10%) were the two most frequent sources for bacteraemia, with the origin remaining unknown in 62 cases (21%). The overall mortality rate was 14%. Further studies focusing on the antimicrobial susceptibility and demographic background of patients are needed to further objectify the currently observed trends.

Carbapenem resistance in Bacteroides fragilis is associated with cfiA -encoded class B metallo-beta-lactamase. cfiA -negative and cfiA -positive isolates belong to genotypically distinct groups. Of a total of 248 B. fragilis isolates included in this study, 214 were susceptible, 10 were intermediate, and 24 were resistant to meropenem. We show that matrix-assisted laser desorption ionization–time of flight mass spectrometry is able to differentiate between cfiA -negative and cfiA -positive isolates and predict carbapenem resistance in a routine laboratory setting.

Recently new resin-based BacT/Alert FAN Plus bottles containing antibiotic-binding polymeric beads, were introduced as an improvement of the charcoal-based FAN bottles for the recovery of bacteria and fungi. To assess the performance of the novel anaerobic FN plus bottles in the detection of anaerobic organisms, we compared the detection rate and the time to detection (TTD) in spiked resin-based FN Plus bottles and charcoal-based FN bottles. The bottles were experimentally seeded with reference strains or clinical strains collected from positive blood cultures. Five reference strains and fifty-five clinically significant anaerobic isolates were investigated, of which 91% (61/67) showed growth in both types of bottles within an incubation time of 5 days. A significant prolonged median TTD of 45 h for anaerobic microorganisms was observed in the resin-based bottles versus 29 h in the charcoal-based bottles (P

Background Fusobacterium spp. from clinical specimens are increasingly reported. We sought to describe the epidemiology, the microbiological, and the clinical characteristics of head and neck infections caused by Fusobacterium necrophorum and other Fusobacterium spp. Materials and methods Retrospective cohort study between October 1st, 2004 and September 30th, 2014 performed in an academic hospital. Electronic patient charts and the laboratory information system were reviewed for demographic and microbiological data. The number and percentages of specific diagnosis and treatment among patients with positive Fusobacterium spp. culture were calculated. The incidence was calculated based on the number of specimens investigated each year. Results Included were 230 cultures of 230 patients (median age of 28 years, 61.7% men). F. necrophorum was often found in young patients with high C-reactive protein (CRP) and high number of leukocytes in blood. Other Fusobacterium spp. were often found in middle aged patients with relatively high CRP and slightly increased leukocytes. Three major causes of the isolation of F. necrophorum and other Fusobacterium spp. were acute tonsillitis (n = 18, incidence of 0.2%), peritonsillar abscess (n = 39, 0.5%) and acute otitis (n = 45, 0.1%). While F. necrophorum was found in majority (37/57) of patients with acute tonsillitis or peritonsillar abscess, Fusobacterium spp. other than F. necrophorum were found in the majority (35/45) of patients with acute otitis. Isolated fusobacteria were susceptible to beta-lactam antibiotics, clindamycin and metronidazole. The outcomes of patients with Fusobacterium spp. were good. Conclusion F. necrophorum and other Fusobacterium spp. are rare cause of head and neck infections. The infections are well treated by combination of antibiotics and surgery.

Reptile-associated salmonellosis in humans is an increasing public health issue. This study aimed at characterizing Salmonella isolates from captive lizards and to compare them to human isolates. Salmonella was isolated from 25 of 33 cloacal and 47 of 79 faecal samples from captive lizards (75.8 and 59.5%, respectively). The strains belonged to 44 serotypes of subspecies I (27 serotypes), II (9), IIIb (3) and IV (5). Two strains, one of serotype Enteritidis and one of serotype Amsterdam, were resistant to nitrofurantoin. Invasion assays in Caco-2 cells were performed with 40 saurian isolates of subspecies I, 15 isolates of subspecies II, 4 strains of subspecies IIIb, 6 subspecies IV isolates and 17 human isolates of corresponding serotypes of subspecies I. Saurian isolates belonging to subspecies I invaded the Caco-2 cells to a higher extent than those from the other subspecies. The human isolates invaded the Caco-2 cells to a lesser degree compared to their saurian counterparts. In the same strains, the presence of virulence genes agfA, shdA, spvR, pefA and sopE was determined using PCR. Whereas agfA was detected in all strains, pefA was only detected in one saurian and in the human serotype Enteritidis strains. The spvR gene was detected in the same serotype Enteritidis strains and in 33% of the subspecies IV strains. The shdA gene was present in all the human isolates and in 86% of subspecies I saurian isolates. SopE was found in 17% of the human isolates, in 24% of the saurian subspecies I strains and in all of the subspecies IV strains.

No abstract available

L'ertapeneme est un nouveau carbapeneme a usage parenteral avec une demi-vie serique prolongee. Son spectre d'activite est similaire a celui de l'imipeneme et du meropeneme a l'egard des bacteries a Gram positif, des enterobacteries, et des bacilles a Gram negatif fastidieux mais son activite est moindre sur Pseudomonas aeruginosa et Acinetobacter spp. Plusieurs etudes americaines mais seulement une etude europeenne ont montre que l'ertapeneme a une excellente activite sur les anaerobies. Les objectifs de cette etude etaient de tester l'activite de l'ertapeneme vis-a-vis d'anaerobies isoles de facon prospective des tractus respiratoires bas et haut, et de comparer leur susceptibilite a celle d'isolats anaerobies provenant d'autres sites. Cinquante-trois (53) isolats du tractus respiratoire ainsi que 50 isolats d'autres sites corporels ont ete testes a l'aide d'E-tests pour 6 antibiotiques. Pour les isolats respiratoires et ceux d'autres sites les CMI 90 (mg/L) etaient respectivement > 32 et > 32 pour la penicilline, 0,38 et 0,75 pour l'association amoxicilline-clavulanate, 48 et > 256 pour la ceftriaxone, 0,12 et 0,75 pour l'ertapeneme, 12 et > 256 pour la clindamycine et 2 et 12 pour la moxifloxacine. La sensibilite superieure des isolats respiratoires etait principalement due a la distribution differente des especes isolees: seulement 3 isolats respiratoires mais 22 autres isolats appartenaient au groupe de Bacteroides fragilis. Cette etude confirme l'excellente activite de l'ertapeneme vis-a-vis des anaerobies du tractus respiratoire.

Corynebacterium diphtheriae and Corynebacterium ulcerans are rarely isolated from clinical samples in Belgium. A case of toxigenic C. ulcerans in a woman is described, which confirms that this pathogen is still present. During investigation of the patient's cats, only a non-toxigenic toxin-bearing C. diphtheriae strain was detected.

Eight coagulase-negative, novobiocin-susceptible staphylococcal strains were isolated from human clinical specimens at two different Belgian medical facilities. All strains were non-motile, Gram-stain-positive, catalase-positive cocci. DNA G+C content, peptidoglycan type, menaquinone pattern, the presence of teichoic acid and cellular fatty acid composition were in agreement with the characteristics of species of the genusStaphylococcus. Sequencing of the 16S rRNA gene and four housekeeping genes (dnaJ,tuf,gapandrpoB) demonstrated that these strains constitute a separate taxon within the genusStaphylococcus. Less than 41 % DNA–DNA hybridization with the most closely related species of the genusStaphylococcus(Staphylococcus haemolyticus,Staphylococcus hominisandStaphlococcus lugdunensis) was observed. Key biochemical characteristics that allowed these bacteria to be distinguished from their nearest phylogenetic neighbours are arginine dihydrolase positivity, ornithine decarboxylase negativity and inability to produce acid aerobically fromd-mannose, α-lactose and turanose. Acid is produced aerobically from trehalose. Based on these results, a novel species of the genusStaphylococcusis described and namedStaphylococcusjettensissp. nov. The type strain is SEQ110T( = LMG 26879T = CCUG 62657T = DSM 26618T).

Sequence-based typing (SBT) is a discriminatory method widely used to genotype Legionella pneumophila strains. A total of 86 clinical L. pneumophila serogroup 1 (sg1) isolates, collected between January 2000 and December 2010 in the two Belgian National Reference Centres for Legionella pneumophila, were genotyped using the internationally standardised SBT protocol of the European Working Group for Legionella Infections (EWGLI). The isolates could be classified into 31 different sequence types (ST, index of diversity: 0.879). The obtained STs were submitted to the EWGLI SBT-database for L. pneumophila. In our study, ST47 (27.9%) and ST1 (19.8%) were the most frequently detected STs. The detected profiles were a combination of both frequently isolated and unique STs, and of both worldwide distributed and more local strains. Two STs, ST880 and ST881, were new to the EWGLI database. In conclusion, we characterised L. pneumophila sg1 isolates with the SBT method, and created a Belgian profile database that will be useful for future epidemiological studies.

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During the 2009 influenza A/H1N1v pandemic, children were identified as a specific "at risk" group. We conducted a multicentric study to describe pattern of influenza A/H1N1v infection among hospitalized children in Brussels, Belgium., Journal Article, SCOPUS: ar.j, info:eu-repo/semantics/published

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As we have been users of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) since January 2009, we read with much interest the paper of van Veen et al. ([9][1]) in a recent issue of the Journal of Clinical Microbiology. We would like to respond to two issues

BACKGROUND: Extraintestinal Enterobius vermicularis infections are rare but may occasionally affect the female genital tract. Although mostly asymptomatic or causing minor clinical problems, they may lead to severe infectious complications. METHODS: Case report and review of the pertinent English language literature. RESULTS: A 31-year-old, 30-week-pregnant female was admitted with a clinical suspicion of appendicitis. At surgery, the appendix appeared normal, but generalized peritonitis of unclear origin was present. Eggs of Enterobius vermicularis were found upon microbiological and pathological examination. Because of persisting infectious disease, the patient underwent an elective caesarean section, and at that time the diagnosis of a right tuboovarian abscess was made, and salpingo-oophorectomy was performed. The pathology report confirmed the diagnosis of an E. vermicularis salpingo-oophoritis. CONCLUSION: This case was extraordinary because of a combination of tuboovarian abscess and generalized peritonitis with E. vermicularis infection occurring during late pregnancy. Ectopic enterobiasis should be considered in the differential diagnosis of pelvic infections of gynecological origin.

Objectives: To collect recent data on the susceptibility of anaerobes and to compare them with results from previous studies. Methods: Four hundred and forty-three anaerobic clinical isolates from various body sites were prospectively collected from October 2003 to February 2005 in nine Belgian hospitals. MICs were determined for nine anti-anaerobic and three recently developed antibiotics. Results: Most Gram-negative bacilli except Fusobacterium spp. were resistant to penicillin. Piperacillin/tazobactam, metronidazole, chloramphenicol, meropenem and amoxicillin/clavulanic acid were very active against all groups, but only 86% of Bacteroides fragilis group strains were susceptible to the latter. Cefoxitin, cefotetan and clindamycin were less active. In particular, only 62%, 52% and 48% of B. fragilis group strains were susceptible, respectively. Clindamycin shows a continuing decrease in activity, as 83% were still susceptible in 1987 and 66% in 1993-94. Anti-anaerobic activity of the new antibiotics is interesting, with MIC50 and MIC90 of 1 and > 32 mg/L for moxifloxacin, 2 and 4 mg/L for linezolid and 0.5 and 8 mg/L for tigecycline. Conclusions: The susceptibility of anaerobic bacteria remains stable in Belgium, except for clindamycin, which shows a continuous decrease in activity. However, for each of the tested antibiotics, at least a few resistant organisms were detected. Consequently, for severe infections involving anaerobic bacteria, it could be advisable to perform microbiological testing instead of relying on known susceptibility profiles. Periodically monitoring background susceptibility remains necessary to guide empirical therapy.

Carbapenem-resistant Acinetobacter baumannii isolates were obtained from 17 patients between September 2004 and August 2005 at the Academisch Ziekenhuis Vrije Universiteit Brussel, Brussels, Belgium. These multidrug-resistant isolates, which belonged to a single clone, remained susceptible to colistin and tigecycline only and produced the carbapenem-hydrolyzing oxacillinase OXA-58. This study highlights the importance of the intercountry spread of this β-lactamase-mediated resistance mechanism and its epidemic evolution.

Reptile-associated salmonellosis in humans is an increasing public health issue. This study aimed at characterizing Salmonella isolates from captive lizards and to compare them to human isolates. Salmonella was isolated from 25 of 33 cloacal and 47 of 79 faecal samples from captive lizards (75.8 and 59.5%, respectively). The strains belonged to 44 serotypes of subspecies I (27 serotypes), II (9), IIIb (3) and IV (5). Two strains, one of serotype Enteritidis and one of serotype Amsterdam, were resistant to nitrofurantoin. Invasion assays in Caco-2 cells were performed with 40 saurian isolates of subspecies I, 15 isolates of subspecies II, 4 strains of subspecies IIIb, 6 subspecies IV isolates and 17 human isolates of corresponding serotypes of subspecies I. Saurian isolates belonging to subspecies I invaded the Caco-2 cells to a higher extent than those from the other subspecies. The human isolates invaded the Caco-2 cells to a lesser degree compared to their saurian counterparts. In the same strains, the presence of virulence genes agfA, shdA, spvR, pefA and sopE was determined using PCR. Whereas agfA was detected in all strains, pefA was only detected in one saurian and in the human serotype Enteritidis strains. The spvR gene was detected in the same serotype Enteritidis strains and in 33% of the subspecies IV strains. The shdA gene was present in all the human isolates and in 86% of subspecies I saurian isolates. SopE was found in 17% of the human isolates, in 24% of the saurian subspecies I strains and in all of the subspecies IV strains.

The genus Arcobacter currently includes four species ([7][1]). Two species, Arcobacter cryaerophilus and Arcobacter butzleri , have been associated with human disease. They were mainly isolated from patients with diarrhea and bacteremia ([2][2], [3][3], [6][4]). Arcobacter nitrofigilis has been