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The objectives of this study were to assess Candida spp. distribution and antifungal resistance of candidaemia across Europe. Isolates were collected as part of the third ECMM Candida European multicentre observational study, conducted from 01 to 07-07-2018 to 31-03-2022. Each centre (maximum number/country determined by population size) included ∼10 consecutive cases. Isolates were referred to central laboratories and identified by morphology and MALDI-TOF, supplemented by ITS-sequencing when needed. EUCAST MICs were determined for five antifungals. fks sequencing was performed for echinocandin resistant isolates. The 399 isolates from 41 centres in 17 countries included C. albicans (47.1%), C. glabrata (22.3%), C. parapsilosis (15.0%), C. tropicalis (6.3%), C. dubliniensis and C. krusei (2.3% each) and other species (4.8%). Austria had the highest C. albicans proportion (77%), Czech Republic, France and UK the highest C. glabrata proportions (25–33%) while Italy and Turkey had the highest C. parapsilosis proportions (24–26%). All isolates were amphotericin B susceptible. Fluconazole resistance was found in 4% C. tropicalis , 12% C. glabrata (from six countries across Europe), 17% C. parapsilosis (from Greece, Italy, and Turkey) and 20% other Candida spp. Four isolates were anidulafungin and micafungin resistant/non-wild-type and five resistant to micafungin only. Three/3 and 2/5 of these were sequenced and harboured fks -alterations including a novel L657W in C. parapsilosis. The epidemiology varied among centres and countries. Acquired echinocandin resistance was rare but included differential susceptibility to anidulafungin and micafungin, and resistant C. parapsilosis. Fluconazole and voriconazole cross-resistance was common in C. glabrata and C. parapsilosis but with different geographical prevalence. • Candidaemia epidemiology differs notably across 41 centres in 17 countries. • C. albicans proportions varied from 77% in Austria to 36% in Turkey. • Acquired fluconazole resistance emerges in C. parapsilosis in southern Europe. • A novel echinocandin resistance mutation and MDR was demonstrated in C. parapsilosis. • Antifungal stewardship and infection control is of utmost importance. [ABSTRACT FROM AUTHOR]

Dendritic cells (DC) are antigen-presenting cells (APC) that most efficiently initiate and control immune responses. Migration processes of blood DC are crucial to exert their professional antigen-presenting functions. Matrix-degrading metalloproteinases (MMP) are proteolytic enzymes, which are considered to be key enzymes in extracellular matrix (ECM) turnover and mediators of cell migration. Tissue inhibitors of metalloproteinases (TIMP) are important regulators of MMP activity. Here we investigate whether blood monocyte-derived immature DC (iDC) and mature DC (mDC) express, produce and secrete functionally active MMP-1, -2, -3 and -9 and their inhibitors TIMP-1 and -2, and examine their involvement in multiple sclerosis (MS). On mRNA level, we observed high numbers of MMP-2 and TIMP-2 mRNA expressing iDC in MS. On protein level, high percentages of MMP-1, -2 and -9 expressing iDC by flow cytometry, and high MMP-1 secretion by Western blot together with high MMP-2 and -9 activities in iDC supernatants as studied with zymography were observed. Similarly, MS is associated with high percentages of MMP-2 and -3 and of TIMP-1 expressing mDC by flow cytometry together with high MMP-3 secretion and high MMP-9 activity in culture supernatants. Spontaneous migratory capacity of both iDC and mDC over ECM-coated filters was higher in MS compared to healthy controls (HC).In conclusion, blood monocyte-derived iDC and mDC express, produce and secrete several MMP and TIMP. Alterations in these molecules as observed in MS may be functionally important for DC functioning. [Copyright &y& Elsevier]

The European Confederation of Medical Mycology (ECMM) collected data on epidemiology, risk factors, treatment, and outcomes of patients with culture-proven candidaemia across Europe to assess how adherence to guideline recommendations is associated with outcomes. In this observational cohort study, 64 participating hospitals located in 20 European countries, with the number of eligible hospitals per country determined by population size, included the first ten consecutive adults with culture-proven candidaemia after July 1, 2018, and entered data into the ECMM Candida Registry (FungiScope Candi Reg). We assessed ECMM Quality of Clinical Candidaemia Management (EQUAL Candida) scores reflecting adherence to recommendations of the European Society of Clinical Microbiology and Infectious Diseases and the Infectious Diseases Society of America guidelines. 632 patients with candidaemia were included from 64 institutions. Overall 90-day mortality was 43% (265/617), and increasing age, intensive care unit admission, point increases in the Charlson comorbidity index score, and Candida tropicalis as causative pathogen were independent baseline predictors of mortality in Cox regression analysis. EQUAL Candida score remained an independent predictor of mortality in the multivariable Cox regression analyses after adjusting for the baseline predictors, even after restricting the analysis to patients who survived for more than 7 days after diagnosis (adjusted hazard ratio 1·08 [95% CI 1·04–1·11; p<0·0001] in patients with a central venous catheter and 1·09 [1·05–1·13; p<0·0001] in those without one, per one score point decrease). Median duration of hospital stay was 15 days (IQR 4–30) after diagnosis of candidaemia and was extended specifically for completion of parenteral therapy in 100 (16%) of 621 patients. Initial echinocandin treatment was associated with lower overall mortality and longer duration of hospital stay among survivors than treatment with other antifungals. Although overall mortality in patients with candidaemia was high, our study indicates that adherence to clinical guideline recommendations, reflected by higher EQUAL Candida scores, might increase survival. New antifungals, with similar activity as current echinocandins but with longer half-lives or oral bioavailability, are needed to reduce duration of hospital stay. Scynexis. [ABSTRACT FROM AUTHOR]

Short-term culture followed by MALDI-TOF MS is one of the most widely used methods for fast identification of microorganisms from blood cultures. The method identifies the vast majority of bloodstream infection pathogens in 2–6 h after positive blood culture. Transport time of blood culture bottles to laboratories is a major problem affecting total turnaround time. Therefore, many central laboratories establish satellite blood culture systems in other clinics and hospitals to allow blood culture bottles to be incubated immediately after sampling. However, positive blood culture bottles still need to be transported to the clinical microbiology laboratory for analysis. The aim of this study was to investigate how delayed analysis of positive blood culture bottles would affect the short-term culture followed by MALDI-TOF MS method. To simulate the effect of transportation and delayed analysis of blood culture bottles, 51 simulated blood culture bottles were incubated for 0, 2, 4 and 24 h at room temperature. After each time interval, a 2 to 4 h short-term culture followed by MALDI-TOF MS was performed. In addition, 257 prospective clinical positive blood culture bottles were analysed with the same method after a 24 h incubation at room temperature. In simulated samples, all (120/120) Gram-negative bacteria and 77/84 (91.6%) Gram-positive bacteria were accurately identified at species-level after a 2 h short-term culture, regardless of the duration of simulated transport time. In the clinical samples, 100/116 (86.2%) Gram-negative, and 44/141 (31.2%) Gram-positive bacteria were accurately identified at species-level after a 2 h short-term culture. After contaminants were excluded, 39/71 (54.9%) Gram-positive bacteria could be identified after 2 h. After a 4 h short-term culture, 112/116 (96.6%) Gram-negative, and 108/141 (76.6%) Gram-positive bacteria were accurately identified at species-level. Of the clinically relevant Gram-positive bacteria, 68/71 (95.8%) were identified at species-level after 4 h. Short-term culture followed by MALDI-TOF MS can provide fast and accurate results for identification of clinically relevant bacteria, despite long transportation times from satellite laboratories. The present data shows that the method can be used for identification of microorganisms from positive blood cultures transported from satellite blood culture systems. • Impact of transport time on rapid identification of positive blood cultures. • Most bacteria were reliably identified despite long transport times. • Gram-negative bacteria were identified more reliably than Gram-positive bacteria. [ABSTRACT FROM AUTHOR]

[Display omitted] • This study provided the first species-level characterization of OFIs in HNC patients receiving RT using MALDI-TOF MS technology. • Five species including C. albicans , C. glabrata , P. kudriavzevii , C. parapsilosis , and C. tropicalis , were commonly detected in both ongoing- and post-RT patients and appeared to be the core mycobiota of OFIs in HNC. • HNC patients under ongoing-RT had more complex co-occurrence of fungal species than post-RT patients. • Despite the newly emergence of F. solani and C. jadinii , C. albicans remained the most prevalent OFIs pathogen in HNC patients. Head and neck cancer (HNC) impairs patient immunity and increases susceptibility to oral fungal infections (OFIs). Effectively treating such infections requires accurate identification of the causative pathogens. This study aimed to characterize the mycobiota profile of OFIs in HNC patients undergoing radiation treatment (RT). A 6-year retrospective analysis of oral mucosal samples from HNC patients with a history of RT and OFIs between 2014 and 2019 was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) profiling. Samples from the Clinical Microbiology Laboratory at Karolinska University Hospital were evaluated for mycobiota diversity and species co-occurrence patterns in the ongoing-RT and post-RT groups. A total of 190 oral fungi (88% Candida , 5% Pichia) were isolated from 162 HNC patients receiving RT. In the ongoing-RT group, the emergent non-albicans Candida (NAC) species; F. solani and C. jadinii , were detected for the first time. The dominant pathogens in both ongoing and post-RT groups were C. albicans , C. glabrata , P. kudriavzevii , C. parapsilosis , and C. tropicalis , as shown by Venn analysis. Network analysis revealed greater fungi diversity and multi-species co-occurrence in the ongoing-RT group. C. albicans commonly co-occurred with C. glabrata in both ongoing-RT (21%) and post-RT groups (30%). MALDI-TOF MS identified a wide range of oral fungal species in HNC patients receiving RT. While C. albicans remains the most prevalent OFIs pathogen, multi-species co-occurrence and novel NACs were noted. Understanding the ecological interactions among these causative pathogens could significantly advance the development of effective therapeutics for treating OFIs in HNC patients. [ABSTRACT FROM AUTHOR]

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and chromogenic media are widely used in clinical microbiology laboratories to facilitate the rapid selection and identification of pathogens. The aim of this study was to evaluate whether usage of chromogenic media limits the diagnostic performance of MALDI-TOF MS for microbial identification. A total of 386 microorganisms collected and analyzed at five laboratories were included. Isolates were cultured on relevant chromogenic media and non-selective agar plates in parallel and identified using the Bruker MALDI-TOF MS. Among the tested isolates, no misidentification was recorded and there was no medium-related difference in the identification level. However, score values were overall slightly but significantly lower for isolates grown on chromogenic media. In conclusion, the use of chromogenic culture media tested here had no relevant impact on MALDI-TOF MS performance for diagnostic purposes. [ABSTRACT FROM AUTHOR]

Abstract: Granulicatella and Abiotrophia spp. were known as nutritionally variant streptococci (NVS). Such strains have caused major diagnostic difficulties due to fastidious culturing and unspecific colony morphology. The present study is aimed at comparing the performance of laboratory available diagnostic methods for NVS isolates and determining the antimicrobial susceptibility of these isolates. Fourteen clinical invasive isolates, consisting of 10 Granulicatella adiacens, 1 Granulicatella elegans, and 3 Abiotrophia defectiva were in parallel analyzed by 2 matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) systems, i.e., Bruker MS and Vitek MS, as well as Vitek 2 for the species determination. 16S rRNA gene sequencing was applied as a reference method. The Vitek MS gave correct identification for all 14 isolates. The Bruker MS could correctly identify 8/10 G. adiacens, 0/1 G. elegans, and 3/3 A. defectiva isolates at the first analysis occasion, and all 14 isolates became identifiable after repeated tests. The Vitek 2 system could identify 6/10 G. adiacens, 1/1 G. elegans, and 2/3 A. defectiva isolates at the species level. Antimicrobial susceptibilities of 11 antibiotics were determined by Etest. Resistance against ciprofloxacin, ceftriaxone, rifampicin, and tetracycline were observed in 4, 10, 4, and 1 isolates, respectively. In conclusion, MALDI-TOF MS is a useful tool for the rapid diagnosis of NVS. Phenotypic testing by Vitek 2 is only partially effective for the accurate identification of such strains. The emergence of resistant NVS isolates indicates the necessity of monitoring antimicrobial susceptibilities of such uncommon pathogens. [Copyright &y& Elsevier]

Abstract: The purpose of this investigation was to evaluate the performance of 4 supplements: horse blood, fastidious organisms supplement (FOS), haemin isovitalex albumine (HIA), and brain heart infusion–haemin isovitalex albumine (BHI-HIA) and 5 blood culture bottles: Bactec Mycosis IC/F, Plus Aerobic/F, Peds Plus/F from the Bactec 9240 system, and BacT/Alert FA and BacT/Alert PF from the BacT/Alert 3D system, in detection of bacteria and Candida spp. in simulated sterile body fluids other than blood models. In total, 8 reference strains (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, Listeria monocytogenes, Candida albicans, and Candida parapsilosis) and 11 clinical bacteria and yeast isolates (6 isolates from cerebrospinal fluid and 5 isolates from blood) were included in this study. Horse blood, FOS, and HIA were significantly better than no supplements (P < 0.0001, P < 0.0002, and P = 0.05, respectively) in detection of bacteria. Interestingly, there was no significant difference between BHI-HIA and bottles without any supplements. Sixty bottles analyzed of which 59 (98.33%) bottles with horse blood, 53 (88.33%) with FOS, 45 (75.00%) with HIA, and 43 (71.67%) with BHI-HIA signaled positive. The positivity rates with horse blood were significantly higher than with HIA and BHI-HIA (P < 0.0005 and P < 0.0001, respectively). Similarly, the blood culture bottles with horse blood had shorter time to detection (TTD) compared to bottles with FOS and HIA (P < 0.05 and P < 0.0001, respectively). When yeasts were analyzed, almost all (124/125) blood culture bottles with Candida spp. signaled positive even in the absence of supplements. Bactec Mycosis IC/F had significantly shorter TTD compared to Bactec Peds Plus/F, Bactec Plus Aerobic/F, BacT/Alert FA, and BacT/Alert PF bottles in detection of Candida spp. (P < 0.005, P < 0.05, P < 0.001, and P < 0.001, respectively). The present study showed that horse blood was the most effective supplement in growth of bacteria in the blood culture bottles that were analyzed in the study. [Copyright &y& Elsevier]

Abstract: In the present study we investigated transport times for blood cultures from three tertiary-care hospitals to Karolinska University Laboratory and identified predictors of long transport times. Concomitantly, consequences of delayed incubation on total detection time (TDT) were analyzed by in vitro sepsis models. A total of 909 blood cultures were studied. The median (interquartile range) transport time was 9 (3–15) h. The hospital accommodating the microbiology laboratory had the shortest transport time compared to the other two hospitals (P < 0.0001). Samples taken between 16:00–24:00 had longer transport times compared to samples taken between 8:00–16:00 and 24:00–08:00 (P < 0.0001). In vitro experiments showed that TDT was longer for samples pre-incubated at room temperature (RT) for 19 h compared to the ones pre-incubated for 2 h or 9.5 h (P < 0.0001). In conclusion, off-site location, time of sampling and number of transports per day were related to, and predictors of transport time. [Copyright &y& Elsevier]

Abstract: The present study analyzed the performance of Bactec Mycosis IC/F, BacT/Alert FA, and BacT/Alert FN vials in detection and time to detection (TTD) of Candida spp. in 179 simultaneous blood cultures. The Mycosis IC/F, BacT/Alert FA, and BacT/Alert FN vials could detect Candida spp. in 144 (80.45%) of 179, 149 (83.24%) of 179, and 8 (4.47%) of 179 samples, respectively. With the presence of antifungal therapy, the numbers of positive vials were higher in BacT/Alert FA compared to Mycosis IC/F, 87/99 versus 73/99, respectively (P < 0.05). TTD (SD) for C. albicans was shorter in Mycosis IC/F than in BacT/Alert FA vials without antifungal therapy, 20.89 (9.33) versus 28.26 (9.77), respectively (P < 0.01). The detection of Candida spp., with concomitant bacteremia, was higher in Mycosis IC/F than in BacT/Alert FA vials, 28/30 and 19/30, respectively (P = 0.01). The present data show that the use of Bactec Mycosis IC/F together with BacT/Alert FA vials might improve the detection of Candida spp. [Copyright &y& Elsevier]

Abstract: The aim of this study was to compare the performance of four methods that are widely used in the clinical microbiology laboratory for identification of Pasteurella species. The 4 methods evaluated were VITEK2, VITEK MS (BioMerieux), and Bruker Biotyper MS (Bruker) as well as traditional biochemical tests. Sequencing of the sodA gene was used as the reference method. Sixty-five isolates of Pasteurella spp. from 65 patients were analyzed. One Pasteurella multocida isolate from American Type Culture Collection (Manassas, VA, USA) was used as a reference. Traditional biochemical tests accurately identified 62/66 (94%) isolates. Both Bruker and Vitek matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) identified 59/66 (89%) strains, but VITEK2 could only identify 32/66 (48.5%) isolates correctly. The mean time to identification using biochemical tests was 20 hours; VITEK2 took 6 hours and MALDI-TOF approximately 10 minutes. In conclusion, MALDI-TOF is a quick method, which accurately identified most isolates of Pasteurella to the species level. Thus, MALDI-TOF constitutes a valuable diagnostic tool in the clinical laboratory. [Copyright &y& Elsevier]