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Objectives To study the presence of viral RNA in follicular fluid, cumulus cells and endometrial tissue samples in SARS-CoV-2 positive women undergoing assisted reproductive technologies (ART). Design Prospective, single-center, observational study. Setting Tertiary hospital, University Hospital UZ Brussel, Belgium. Patients Sixteen patients undergoing transvaginal oocyte retrieval who had a positive SARS-CoV-2 RNA test less than 48 hours before the procedure. All cases were done between September 2020 and June 2021 and used in-vitro fertilization or intracytoplasmic sperm injection. All embryos were vitrified to avoid conception during SARS-CoV-2 infection. Intervention Follicular fluid aspirated during oocyte retrieval, cumulus cells and endometrial samples were analyzed for SARS-CoV-2 RNA using RealStar® SARS-CoV-2 RT-PCR-Kit1.0 (Altona-Diagnostics). Main outcome measures The primary outcome parameter was the detection of viral RNA in follicular fluid, cumulus cells and endometrial cells. Fertilization rate, embryo developmental potential and clinical outcome after frozen embryo transfer were secondary outcome parameters. Results Samples from sixteen patients were analyzed. Cycle threshold (Ct) values < 40 were considered positive. All samples were negative for SARS-CoV-2 viral RNA. No inflammatory lesions of the endometrium were identified histologically. Fertilization rate, embryo development and clinical outcomes after embryo transfer were reassuring. Conclusions In women infected with SARS-CoV-2 who underwent ART, viral RNA was undetectable in follicular fluid, cumulus cells and endometrium. Caution is warranted in view of the small sample size, and the risk of SARS-CoV-2 affecting the embryo via ART cannot be ruled out. Adequate counselling of women and couples undergoing ART is crucial in parallel with further research on the effect of exposure of the early human embryo to SARS-CoV-2., In patients with a SARS-CoV-2 infection, viral RNA was undetectable in follicular fluid, cumulus cells and endometrial biopsies. Additionally, embryological data and clinical outcomes are reported.

Among Shiga toxin (Stx)-producing Escherichia coli (STEC) strains of various serotypes, O157:H7 and five major non-O157 STEC (O26:H11, O111:H8, O103:H2, O121:H19 and O145:H28) can be selectively isolated by using tellurite-containing media. While human infections by O165:H25 STEC strains have been reported worldwide, their detection and isolation are not easy, as they are not resistant to tellurite. Systematic whole-genome sequencing (WGS) analyses have not yet been conducted. Here, we defined O165:H25 strains and their close relatives, including O172:H25 strains, as clonal complex 119 (CC119) and performed a global WGS analysis of the major lineage of CC119, called CC119 sensu stricto (CC119ss), by using 202 CC119ss strains, including 90 strains sequenced in this study. Detailed comparisons of 13 closed genomes, including 7 obtained in this study, and systematic analyses of Stx phage genomes in 50 strains covering the entire CC119ss lineage, were also conducted. These analyses revealed that the Stx2a phage, the locus of enterocyte effacement (LEE) encoding a type III secretion system (T3SS), many prophages encoding T3SS effectors, and the virulence plasmid were acquired by the common ancestor of CC119ss and have been stably maintained in this lineage, while unusual exchanges of Stx1a and Stx2c phages were found at a single integration site. Although the genome sequences of Stx2a phages were highly conserved, CC119ss strains exhibited notable variation in Stx2 production levels. Further analyses revealed the lack of SpLE1-like elements carrying the tellurite resistance genes in CC119ss and defects in rhamnose, sucrose, salicin and dulcitol fermentation. The genetic backgrounds underlying these defects were also clarified.

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.

The microbiome of the reproductive tract has been associated with (sub)fertility and it has been suggested that dysbiosis reduces success rates and pregnancy outcomes. The endometrial microbiome is of particular interest given the potential impact on the embryo implantation. To date, all endometrial microbiome studies have applied a metagenomics approach. A sequencing-based technique, however, has its limitations, more specifically in adequately exploring low-biomass settings, such as intra-uterine/endometrial samples. In this proof-of-concept study, we demonstrate the applicability of culturomics, a high-throughput culturing approach, to investigate the endometrial microbiome. Ten subfertile women undergoing diagnostic hysteroscopy and endometrial biopsy, as part of their routine work-up at Brussels IVF, were included after their informed consent. Biopsies were used to culture microbiota for up to 30 days in multiple aerobic and anaerobic conditions. Subsequent WASPLab®-assisted culturomics enabled a standardized methodology. Matrix-assisted laser desorption/ionization–time of flight mass spectrometry (MALDI-TOF MS) or 16S rRNA sequencing was applied to identify all of bacterial and fungal isolates. Eighty-three bacterial and two fungal species were identified. The detected species were in concordance with previously published metagenomics-based endometrial microbiota analyses as 77 (91%) of them belonged to previously described genera. Nevertheless, highlighting the added value of culturomics to identify most isolates at the species level, 53 (62.4%) of the identified species were described in the endometrial microbiota for the first time. This study shows the applicability and added value of WASPLab®-assisted culturomics to investigate the low biomass endometrial microbiome at a species level.

Background: Healthcare-associated SARS-CoV-2 infections need to be explored further. Our study is an analysis of hospital-acquired infections (HAIs) and ambulatory healthcare workers (aHCWs) with SARS-CoV-2 across the pandemic in a Belgian university hospital. Methods: We compared HAIs with community-associated infections (CAIs) to identify the factors associated with having an HAI. We then performed a genomic cluster analysis of HAIs and aHCWs. We used this alongside the European Centre for Disease Control (ECDC) case source classifications of an HAI. Results: Between March 2020 and March 2022, 269 patients had an HAI. A lower BMI, a worse frailty index, lower C-reactive protein (CRP), and a higher thrombocyte count as well as death and length of stay were significantly associated with having an HAI. Using those variables to predict HAIs versus CAIs, we obtained a positive predictive value (PPV) of 83.6% and a negative predictive value (NPV) of 82.2%; the area under the ROC was 0.89. Genomic cluster analyses and representations on epicurves and minimal spanning trees delivered further insights into HAI dynamics across different pandemic waves. The genomic data were also compared with the clinical ECDC definitions for HAIs; we found that 90.0% of the ‘definite’, 87.8% of the ‘probable’, and 70.3% of the ‘indeterminate’ HAIs belonged to one of the twenty-two COVID-19 genomic clusters we identified. Conclusions: We propose a novel prediction model for HAIs. In addition, we show that the management of nosocomial outbreaks will benefit from genome sequencing analyses.

Enteropathogenic

As with other pathogens, competitive interactions between Bordetella pertussis strains drive infection risk. Vaccines are thought to perturb strain diversity through shifts in immune pressures; however, this has rarely been measured because of inadequate data and analytical tools. We used 3344 sequences from 23 countries to show that, on average, there are 28.1 transmission chains circulating within a subnational region, with the number of chains strongly associated with host population size. It took 5 to 10 years for B. pertussis to be homogeneously distributed throughout Europe, with the same time frame required for the United States. Increased fitness of pertactin-deficient strains after implementation of acellular vaccines, but reduced fitness otherwise, can explain long-term genotype dynamics. These findings highlight the role of vaccine policy in shifting local diversity of a pathogen that is responsible for 160,000 deaths annually.

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.

Shiga toxin (Stx)-producing Escherichia coli (STEC) are foodborne pathogens causing serious diseases, such as haemorrhagic colitis and haemolytic uraemic syndrome. Although O157:H7 STEC strains have been the most prevalent, incidences of STEC infections by several other serotypes have recently increased. O121:H19 STEC is one of these major non-O157 STECs, but systematic whole genome sequence (WGS) analyses have not yet been conducted on this STEC. Here, we performed a global WGS analysis of 638 O121:H19 strains, including 143 sequenced in this study, and a detailed comparison of 11 complete genomes, including four obtained in this study. By serotype-wide WGS analysis, we found that O121:H19 strains were divided into four lineages, including major and second major lineages (named L1 and L3, respectively), and that the locus of enterocyte effacement (LEE) encoding a type III secretion system (T3SS) was acquired by the common ancestor of O121:H19. Analyses of 11 complete genomes belonging to L1 or L3 revealed remarkable interlineage differences in the prophage pool and prophage-encoded T3SS effector repertoire, independent acquisition of virulence plasmids by the two lineages, and high conservation in the prophage repertoire, including that for Stx2a phages in lineage L1. Further sequence determination of complete Stx2a phage genomes of 49 strains confirmed that Stx2a phages in lineage L1 are highly conserved short-tailed phages, while those in lineage L3 are long-tailed lambda-like phages with notable genomic diversity, suggesting that an Stx2a phage was acquired by the common ancestor of L1 and has been stably maintained. Consistent with these genomic features of Stx2a phages, most lineage L1 strains produced much higher levels of Stx2a than lineage L3 strains. Altogether, this study provides a global phylogenetic overview of O121:H19 STEC and shows the interlineage genomic differences and the highly conserved genomic features of the major lineage within this serotype of STEC.

Two colorimetric broth microdilution antifungal susceptibility tests were compared, Sensititre YeastOne and MICRONAUT-AM for nine antifungal agents. One hundred clinical Candida isolates were tested, representing a realistic population for susceptibility testing in daily practice. The reproducibility characteristics were comparable. Only for fluconazole, caspofungin, 5-flucytosine and amphotericin B, an essential agreement of ≥90% could be demonstrated. Sensititre minimal inhibitory concentrations (MICs) were systematically higher than MICRONAUT MICs for all antifungals, except for itraconazole. CLSI clinical breakpoints (CBPs) and epidemiological cut-off values (ECVs) were used for Sensititre MICs while for MICRONAUT the EUCAST CBPs and ECVs were used. Only fluconazole, micafungin, and amphotericin B had a categorical agreement of ≥90%. For fluconazole, micafungin, and amphotericin B the susceptibility proportions were comparable. Susceptibility proportion of posaconazole and voriconazole was higher using the MICRONAUT system. For itraconazole and anidulafungin, the susceptibility proportion was higher using Sensititre. It was not possible to determine the true MIC values or the correctness of a S/I/R result since both commercial systems were validated against a different reference method. These findings show that there is a significant variability in susceptibility pattern and consequently on use of antifungals in daily practice, depending on the choice of commercial system.

Phages and plasmids play important roles in bacterial evolution and diversification. Although many draft genomes have been generated, phage and plasmid genomes are usually fragmented, limiting our understanding of their dynamics. Here, we performed a systematic analysis of 239 draft genomes and 7 complete genomes of Shiga toxin (Stx)-producing Escherichia coli O145:H28, the major virulence factors of which are encoded by prophages (PPs) or plasmids. The results indicated that PPs are more stably maintained than plasmids. A set of ancestrally acquired PPs was well conserved, while various PPs, including Stx phages, were acquired by multiple sublineages. In contrast, gains and losses of a wide range of plasmids have frequently occurred across the O145:H28 lineage, and only the virulence plasmid was well conserved. The different dynamics of PPs and plasmids have differentially impacted the pangenome of O145:H28, with high proportions of PP- and plasmid-associated genes in the variably present and rare gene fractions, respectively. The dynamics of PPs and plasmids have also strongly impacted virulence gene repertoires, such as the highly variable distribution of stx genes and the high conservation of a set of type III secretion effectors, which probably represents the core effectors of O145:H28 and the genes on the virulence plasmid in the entire O145:H28 population. These results provide detailed insights into the dynamics of PPs and plasmids, and show the application of genomic analyses using a large set of draft genomes and appropriately selected complete genomes.

There are different patterns in the COVID-19 outbreak in the general population and amongst nursing home patients. We investigate the time from symptom onset to diagnosis and hospitalization or the length of stay (LoS) in the hospital, and whether there are differences in the population. Sciensano collected information on 14,618 hospitalized patients with COVID-19 admissions from 114 Belgian hospitals between 14 March and 12 June 2020. The distributions of different event times for different patient groups are estimated accounting for interval censoring and right truncation of the time intervals. The time between symptom onset and hospitalization or diagnosis are similar, with median length between symptom onset and hospitalization ranging between 3 and 10.4 days, depending on the age of the patient (longest delay in age group 20&ndash, 60 years) and whether or not the patient lives in a nursing home (additional 2 days for patients from nursing home). The median LoS in hospital varies between 3 and 10.4 days, with the LoS increasing with age. The hospital LoS for patients that recover is shorter for patients living in a nursing home, but the time to death is longer for these patients. Over the course of the first wave, the LoS has decreased.

Highlights • Hydroxychloroquine (HCQ) 2400 mg over 5 days was used in Belgium for COVID-19. • Impact of HCQ on mortality among 8075 patients with COVID-19 was assessed. • Lower mortality in HCQ-treated patients as compared to supportive care. • Lower mortality was irrespective of symptom duration., Hydroxychloroquine (HCQ) has been largely used and investigated as therapy for COVID-19 across various settings at a total dose usually ranging from 2400 mg to 9600 mg. In Belgium, off-label use of low-dose HCQ (total 2400 mg over 5 days) was recommended for hospitalised patients with COVID-19. We conducted a retrospective analysis of in-hospital mortality in the Belgian national COVID-19 hospital surveillance data. Patients treated either with HCQ monotherapy and supportive care (HCQ group) were compared with patients treated with supportive care only (no-HCQ group) using a competing risks proportional hazards regression with discharge alive as competing risk, adjusted for demographic and clinical features with robust standard errors. Of 8075 patients with complete discharge data on 24 May 2020 and diagnosed before 1 May 2020, 4542 received HCQ in monotherapy and 3533 were in the no-HCQ group. Death was reported in 804/4542 (17.7%) and 957/3533 (27.1%), respectively. In the multivariable analysis, mortality was lower in the HCQ group compared with the no-HCQ group [adjusted hazard ratio (aHR) = 0.684, 95% confidence interval (CI) 0.617–0.758]. Compared with the no-HCQ group, mortality in the HCQ group was reduced both in patients diagnosed ≤5 days (n = 3975) and >5 days (n = 3487) after symptom onset [aHR = 0.701 (95% CI 0.617–0.796) and aHR = 0.647 (95% CI 0.525–0.797), respectively]. Compared with supportive care only, low-dose HCQ monotherapy was independently associated with lower mortality in hospitalised patients with COVID-19 diagnosed and treated early or later after symptom onset.

Purpose: Different respiratory sampling methods exist to identify lower airway pathogens in patients with cystic fibrosis (CF), of which bronchoalveolar lavage (BAL), and expectorated sputum are considered the "gold standard." Because BAL cannot be repeated limitless, the diagnosis of lower respiratory tract infections in non-expectorating patients is challenging. Other sampling techniques are nasal swab, cough swab, and induced sputum. The purpose of this study (NCT02363764) was to compare concordance between the microbiological yield of nasal swab, cough swab, and expectorated sputum in expectorating patients; nasal swab, cough swab, and induced sputum in non-expectorating patients; nasal swab, cough swab, induced sputum, and BAL in patients requiring bronchoscopy ("BAL-group"); and to determine the clinical value of cough swab in non-expectorating patients with CF. Methods: Microbiological yield detected by these different sampling techniques was compared between and within 105 expectorating patients, 30 non-expectorating patients and BAL-group (n = 39) in a single CF clinic. Specificity, sensitivity, positive (PPV), and negative (NPV) predictive values were calculated. Results: Overall low sensitivity (6.3-58.0%) and wide-ranging predictive values (0.0-100.0%) indicated that nasal swab was not appropriate to detect lower airway pathogens [Pseudomonas aeruginosa (Pa), Staphylococcus aureus (Sa), and Haemophilus influenzae (Hi)] in all three patient groups. Microbiological yield, specificity, sensitivity, PPV, and NPV of cough swab and induced sputum were largely similar in non-expectorating patients and in BAL-group (except sensitivity (0.0%) of induced sputum for Hi in BAL-group). Calculations for Pa and Hi could not be performed for non-expectorating patients because of low prevalence (n = 2 and n = 3, respectively). In expectorating patients, concordance was found between cough swab and expectorated sputum, except for Hi (sensitivity of 40.0%). Conclusion: Our findings suggest that cough swab might be helpful in detecting the presence of some typical CF pathogens in the lower airways of clinically stable patients with CF. However, in symptomatic patients, who are unable to expectorate and who have a negative cough swab and induced sample, BAL should be performed as it currently remains the "gold standard."

Under the coordination of the European Centre for Disease Prevention and Control (ECDC), the European Legionnaires’ disease Surveillance Network (ELDSNet) conducts surveillance of Legionnaires’ disease (LD) in Europe. Between 2011 and 2015, 29 countries reported 30,532 LD cases to ECDC (28,188 (92.3%) confirmed and 2,344 (7.7%) probable). Four countries (France, Germany, Italy and Spain) accounted for 70.3% of all reported cases, although their combined populations represented only 49.9% of the study population. The age-standardised rate of all cases increased from 0.97 cases/100,000 population in 2011 to 1.30 cases/100,000 population in 2015, corresponding to an annual average increase of 0.09 cases/100,000 population (95%CI 0.02–0.14; p = 0.02). Demographics and infection setting remained unchanged with ca 70% of cases being community-acquired and 80% occurring in people aged 50 years and older. Clinical outcome was known for 23,164 cases, of whom 2,161 (9.3%) died. The overall case fatality ratio decreased steadily from 10.5% in 2011 to 8.1% in 2015, probably reflecting improved reporting completeness. Five countries (Austria, Czech Republic, Germany, Italy, and Norway) had increasing age-standardised LD notification rates over the 2011−15 period, but there was no increase in notification rates in countries where the 2011 rate was below 0.5/100,000 population.

Routine determination of antimicrobial susceptibility of anaerobic clinical isolates is difficult. The E-test is a practical alternative technique that we evaluated while testing clinical isolates in a multicenter study. The susceptibility to 9 antibiotics (penicillin, amoxycillin/clavulanate, ticarcillin/clavulanate, piperacillin/tazobactam, imipenem, cefoxitin, metronidazole, clindamycin, chloramphenicol) of 351 strains belonging to 63 different species was determined by the NCCLS reference agar dilution procedure using Wilkins-Chalgren agar medium with 5% sheep blood and was compared to the E-test performed on the same medium and using manufacturer's recommendations. The MIC values obtained with the E-test were generally one dilution lower than those obtained with the reference technique, 87.1% of the results being within two dilutions. In terms of susceptibility categories, 95.1% agreement was observed with 3.8% minor errors and only 0.5% major and 0.6% very major errors. With some Fusobacterium spp. and Clostridium spp. strains, the E-test was difficult to read or not interpretable because of the presence of growth within the inhibition zone of all beta-lactam antibiotics, representing a trailing phenomenon. We conclude that, if some interpretation difficulties are taken into account, the E-test is a convenient and reliable technique that can be applied in all clinical laboratories. It could be used for the individual testing of important anaerobes in certain clinical situations but cannot yet be considered as a reference technique. Its utility is emphasised by the increased resistance rate against clindamycin and the appearance of a few strains in the B. fragilis group with a reduced susceptibility against metronidazole.

Five direct radio-immunoassay kits for the dosage of 17 beta-estradiol were assessed for their usefulness to monitor the serum values of 17 beta-estradiol in stimulated and natural ovarian cycles. Prerequisites included a turn-around time of 4 hours for an assay of 100 tubes and a range extended to at least 2,000 ng/l. The following parameters were evaluated: characteristics of the standard curve, sensitivity, precision profile, within- and between-assay precision, analytical drift and linearity. We found that the five tested methods are technically suitable to monitor serum 17 beta-estradiol in stimulated cycles but they all lack precision for low 17 beta-estradiol levels as seen in the early follicular phase of natural cycles.

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MICs were performed on 62 streptococci isolated from the blood of neutropenic patients from 1986 to 1992 using the NCCLS broth microdilution procedure. Species isolated were 43 S. mitis, 6 S.pneumoniae, 5 S.sanguis, 1 S.pyogenes, 1 beta-hemolytic streptococcus of the group G, 1 S.adjacens, 1 S.intermedius, 4 not identified isolates (2 alpha-hemolytic, 1 non-hemolytic and 1 nutritionally variant strains). 26% of all strains were resistant or intermediate to penicillin (MIC 90: 2 mg/l) and 89% to norfloxacine (MIC 90: 32 mg/l). All were susceptible to vancomycin. Among cephalosporins, all strains were susceptible to cefotaxime (MIC 90: 1 mg/l), ceftriaxone (MIC 90: 1 mg/l) and cefepime (MIC 90: 2 mg/l) while 19% were R or I to ceftazidime (MIC 90: 16 mg/l). All were susceptible to imipenem (MIC 90: 0.5 mg/l) and meropenem (MIC 90: 1 mg/l). Among the third generation cephalosporins, ceftazidime--the agent of this class of antibiotics that is most often used for the empirical therapy of febrile episodes in neutropenic patients (in combination but also in monotherapy)--has a high resistance rate as compared to the other compounds.

Background Type 1 diabetes mellitus is a T-cell-mediated autoimmune disease that leads to a major loss of insulin-secreting beta cells. The further decline of beta-cell function after clinical onset might be prevented by treatment with CD3 monoclonal antibodies, as suggested by the results of a phase 1 study. To provide proof of this therapeutic principle at the metabolic level, we initiated a phase 2 placebo-controlled trial with a humanized antibody, an aglycosylated human IgG1 antibody directed against CD3 (ChAglyCD3). Methods In a multicenter study, 80 patients with new-onset type 1 diabetes were randomly assigned to receive placebo or ChAglyCD3 for six consecutive days. Patients were followed for 18 months, during which their daily insulin needs and residual beta-cell function were assessed according to glucose-clamp-induced C-peptide release before and after the administration of glucagon. Results At 6, 12, and 18 months, residual beta-cell function was better maintained with ChAglyCD3 than with placebo. The insulin dose increased in the placebo group but not in the ChAglyCD3 group. This effect of ChAglyCD3 was most pronounced among patients with initial residual beta-cell function at or above the 50th percentile of the 80 patients. In this subgroup, the mean insulin dose at 18 months was 0.22 IU per kilogram of body weight per day with ChAglyCD3, as compared with 0.61 IU per kilogram with placebo (P Conclusions Short-term treatment with CD3 antibody preserves residual beta-cell function for at least 18 months in patients with recent-onset type 1 diabetes.

A co-outbreak of verocytotoxin-producing Escherichia coli (VTEC) O145:H28 and O26:H11 infections among participants of two birthday parties in the province of Antwerp (Belgium) was detected in September 2007. Analysis of questionnaires revealed that all cases had consumed ice cream produced at a local dairy farm using pasteurized milk. Combining epidemiological and clinical data, five cases of hemolytic uremic syndrome (HUS) and seven cases of acute diarrhea were identified among the participants. Laboratory investigations were only performed for the HUS patients, and VTEC infection was confirmed in three cases. While both serotypes O145:H28 and O26:H11 were isolated from a two-year-old patient, only VTEC O145:H28 was isolated from an eight-year-old patient, and VTEC O145:H28 infection was confirmed by a positive serology test in a third case (11 years). In order to trace the source of infection, an environmental study was performed. VTEC serotypes O145:H28 and O26:H11 were isolated from leftover ice cream from one of the birthday parties. Quantitative analysis showed a concentration of 2.4 CFU g-1 and 0.03 CFU g-1 of VTEC O145:H28 and O26:H11, respectively. Both serotypes were also detected in fecal samples from calves at the farm. Moreover, VTEC O145:H28 was found in animal feed (hay) and dust present in the positive calves' stable. However, no VTEC was isolated from raw milk. VTEC O145:H28 and O26:H11 isolated from patients and environmental samples respectively, showed indistinguishable patterns after PFGE analysis. PCR revealed that the VTEC O145 and O26 possessed VT2 and VT1 genes, respectively, while both were positive for eaeA and ehxA. It was shown that the VTEC O26 only possessed an incomplete O Island 122 (OI122) gene cluster, while the VTEC O145 contained a complete OI122. To our knowledge, this is the first description of a VTEC O145:H28 carrying a complete OI122 gene cluster. We report a co-outbreak of two non-O157 VTEC serotypes, both belonging to seropathotype B. The lower number of HUS cases with VTEC O26:H11 infections may be due to the lower concentration of VTEC O26:H11 compared to that of VTEC O145:H28 in the ice cream, or to the differences in virulence profiles of these two strains.

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

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