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The aims of this study were to characterize the antifungal activity of amphotericin B against Candida auris in a static in vitro system and to evaluate different dosing schedules and MIC scenarios by means of semi-mechanistic pharmacokinetic/pharmacodynamic (PK/PD) modelling and simulation. A two-compartment model consisting of a drug-susceptible and a drug-resistant subpopulation successfully characterized the time-kill data and a modified E(max) sigmoidal model best described the effect of the drug. The model incorporated growth rate constants for both subpopulations, a death rate constant and a transfer constant between both compartments. Additionally, the model included a parameter to account for the delay in growth in the absence or presence of the drug. Amphotericin B displayed a concentration-dependent fungicidal activity. The developed PK/PD model was able to characterize properly the antifungal activity of amphotericin B against C. auris. Finally, simulation analysis revealed that none of the simulated standard dosing scenarios of 0.6, 1 and 1.5 mg/kg/day over a week treatment showed successful activity against C. auris infection. Simulations also pointed out that an MIC of 1 mg/L would be linked to treatment failure for C. auris invasive infections and therefore, the resistance rate to amphotericin B may be higher than previously reported.

Critically ill COVID-19 patients have higher pro-inflammatory (IL-1, IL-2, IL-6, tumor necrosis alpha) and anti-inflammatory (IL-4, IL-10) cytokine levels, less CD 4 interferon-gamma expression, and fewer CD 4 and CD 8 cells. This severe clinical situation increases the risk of serious fungal infections, such as invasive pulmonary aspergillosis, invasive candidiasis or Pneumocystis jirovecii pneumonia. However, few studies have investigated fungal coinfections in this population. We describe an update on published reports on fungal coinfections and our personal experience in three Spanish hospitals. We can conclude that despite the serious disease caused by SARS-CoV-2 in many patients, the scarcity of invasive mycoses is probably due to the few bronchoscopies and necropsies performed in these patients because of the high risk in aerosol generation. However, the presence of fungal markers in clinically relevant specimens, with the exception of bronchopulmonary colonization by Candida, should make it advisable to early implement antifungal therapy.

Although the Sensititre Yeast-One (SYO) and Etest methods are widely utilized, interpretive criteria are not available for triazole susceptibility testing of Candida or Aspergillus species. We collected fluconazole, itraconazole, posaconazole, and voriconazole SYO and Etest MICs from 39 laboratories representing all continents for (method/agent-dependent) 11,171 Candida albicans, 215 C. dubliniensis, 4,418 C. glabrata species complex, 157 C. guilliermondii (Meyerozyma guilliermondii), 676 C. krusei (Pichia kudriavzevii), 298 C. lusitaniae (Clavispora lusitaniae), 911 C. parapsilosis sensu stricto, 3,691 C. parapsilosis species complex, 36 C. metapsilosis, 110 C. orthopsilosis, 1,854 C. tropicalis, 244 Saccharomyces cerevisiae, 1,409 Aspergillus fumigatus, 389 A. flavus, 130 A. nidulans, 233 A. Niger, and 302 A. terreus complex isolates. SYO/Etest MICs for 282 confirmed non-wild-type (non-WT) isolates were included: ERG11 (C. albicans), ERG11 and MRR1 (C. parapsilosis), cyp51A (A. fumigatus), and CDR2 and CDR1 overexpression (C. albicans and C. glabrata, respectively). Interlaboratory modal agreement was superior by SYO for yeast species and by the Etest for Aspergillus spp. Distributions fulfilling CLSI criteria for epidemiological cutoff value (ECV) definition were pooled, and we proposed SYO ECVs for S. cerevisiae and 9 yeast and 3 Aspergillus species and Etest ECVs for 5 yeast and 4 Aspergillus species. The posaconazole SYO ECV of 0.06 g/ml for C. albicans and the Etest itraconazole ECV of 2 g/ml for A. fumigatus were the best predictors of non-WT isolates. These findings support the need for method-dependent ECVs, as, overall, the SYO appears to perform better for susceptibility testing of yeast species and the Etest appears to perform better for susceptibility testing of Aspergillus spp. Further evaluations should be conducted with more Candida mutants. Copyright © 2018 American Society for Microbiology. All Rights Reserved.

Background: A high quality integrated process in the clinical setting of non-neutropenic critically ill patients at risk for invasive candidiasis is a necessary tool to improve the management of these patients. Aims: To identify the key points on invasive candidiasis in order to develop a set of recommendations with a high level of consensus required for the creation of a total quality integrated process for the management of non-neutropenic critically ill patients at risk of invasive candidiasis. Methods: After a thorough review of the literature of the previous five years, a Spanish prospective questionnaire, which measured consensus by the Delphi technique, was anonymously conducted by e-mail, including 31 national multidisciplinary experts with extensive experience in invasive fungal infections, from six national scientific societies. The experts included a specialist in intensive care medicine, anesthetists, microbiologists, pharmacologists, and specialists in infectious diseases that responded 27 questions prepared by the coordination group. The educational objectives considered six processes that included knowledge of the local epidemiology, the creation and development of multidisciplinary teams, the definitions of the process, protocols, and indicators (KPI), an educational phase, hospital implementation, and the measurement of outcomes. The level of agreement among experts in each category to be selected should exceed 70%. In a second phase, after drawing up the recommendations of the selected processes, a face to face meeting with more than 60 specialists was held. The specialists were asked to validate the pre-selected recommendations. Measures and main outcomes: Firstly, 20 recommendations from all the sections were pre-selected: Knowledge of local epidemiology (3 recommendations), creation and development of multidisciplinary teams (3), definition of the process, protocols and indicators (1), educational phase (3), hospital implementation (3), and measurement of outcomes (7). After the second phase, 18 recommendations were validated, and it was concluded that the minimum team or core necessary for the development of an efficient program in the use of antifungal drugs in non-neutropenic critically ill patients must consist of a specialist in infectious diseases, a clinical pharmacist, a microbiologist, a specialist in intensive care medicine, a specialist in anesthesia and recovery, and an administrator or member of the medical management team, and, in order to be cost-effective, it should be implemented in hospitals with over 200 beds. In addition, it is recommended to apply a consensual check list for the evaluation of the diagnostic process and treatment of invasive candidiasis in patients that have started an antifungal treatment. The management of external knowledge and individual learning stand out as active educational strategies. The main strategies for measuring patient safety outcomes are the analysis of the results achieved, and learning activities; assess, review and refine the deployment of the processes; quality control; epidemiological surveillance and applied research; benchmarking; and basic research. The results of the integrated process should be annually disseminated outside the hospital. Conclusions: Optimizing the management of invasive candidiasis requires the application of the knowledge and skills detailed in our recommendations. These recommendations, based on the Delphi methodology, facilitate the creation of a total quality integrated process in critically-ill patients at risk for invasive candidiasis. (C) 2017 Asociacion Espanola de Micologia. Published by Elsevier Espana, S.L.U. All rights reserved.

Objectives: Infections caused by Candida species are common in critically ill patients and contribute to significant morbidity and mortality. The EPICO Project (Epico 1 and Epico 2.0 studies) recently used a Delphi approach to elaborate guidelines for the diagnosis and treatment of this condition in critically ill adult patients. We aimed to evaluate the impact of a multifaceted educational intervention based on the Epico 1 and Epico 2.0 recommendations. Design: Specialists anonymously responded to two online surveys before and after a multifaceted educational intervention consisting of 60 -min educational sessions, the distribution of slide kits and pocket guides with the recommendations, and an interactive virtual case presented at a teleconference and available for online consultation. Setting: A total of 74 Spanish hospitals. Participants: Specialists of the Intensive Care Units in the participating hospitals. Variables of interest: Specialist knowledge and reported practices evaluated using a survey. The McNemar test was used to compare the responses in the pre- and post -intervention surveys. Results: A total of 255 and 248 specialists completed both surveys, in both periods, respectively. The pre -intervention surveys showed many specialists to be unaware of the best approach for managing invasive candidiasis. After both educational interventions, specialist knowledge and reported practices were found to be more in line with nearly all the recommendations of the Epico 1 and Epico 2.0 guidelines, except as regards de-escalation from echinocandins to fiuconazole in Candida glabrata infections (p= 0.055), and the duration of antifungal treatment in both candidemia and peritoneal candidiasis. Conclusions: This multifaceted educational intervention based on the Epico Project recommendations improved specialist knowledge of the management of invasive candidiasis in critically ill patients. (C) 2016 Elsevier Espana, S.L.U. y SEMICYUC. All rights reserved.

Anidulafungin, caspofungin, and micafungin killing activities against Candida glabrata, Candida bracarensis, and Candida nivariensis were evaluated by the time-kill methodology. The concentrations assayed were 0.06, 0.125, and 0.5 mu g/ml, which are achieved in serum. Anidulafungin and micafungin required between 13 and 26 h to reach the fungicidal endpoint (99.9% killing) against C. glabrata and C. bracarensis. All echinocandins were less active against C. nivariensis.

Candida albicans remains the most common fungal pathogen. This species is closely related to 2 phenotypically similar cryptic species. Candida dubliniensis and Candida africana. This study aims to compare the antifungal activities of echinocandins against 7 Candida albicans, 5 Candida dubliniensis, and 2 Candida africana strains by time-kill methodology. MIC values were similar for the 3 species; however, differences in killing activity were observed among species, isolates, and echinocandins. Echinocandins produced weak killing activity against the 3 species. In all drugs, the fungicidal endpoint (99.9% mortality) was reached at = 0.5 mu g/mL for anidulafungin in 4 Candida albicans and 1 Candida dubliniensis, for caspofungin in 1 Candida albicans and 2 Candida dubliniensis, and for micafungin in 4 Candida albicans and 1 Candida dubliniensis. None of echinocandins showed lethality against Candida africana. Identification of these new cryptic species and time-kill studies would be recommendable when echinocandin treatment fails. (C) 2015 Elsevier Inc. All rights reserved.

Neither breakpoints (BPs) nor epidemiological cutoff values (ECVs) have been established for Candida spp. with anidulafungin, caspofungin, and micafungin when using the Sensititre YeastOne (SYO) broth dilution colorimetric method. In addition, reference caspofungin MICs have so far proven to be unreliable. Candida species wild-type (WT) MIC distributions (for microorganisms in a species/drug combination with no detectable phenotypic resistance) were established for 6,007 Candida albicans, 186 C. dubliniensis, 3,188 C. glabrata complex, 119 C. guilliermondii, 493 C. krusei, 205 C. lusitaniae, 3,136 C. parapsilosis complex, and 1,016 C. tropicalis isolates. SYO MIC data gathered from 38 laboratories in Australia, Canada, Europe, Mexico, New Zealand, South Africa, and the United States were pooled to statistically define SYO ECVs. ECVs for anidulafungin, caspofungin, and micafungin encompassing >97.5% of the statistically modeled population were, respectively, 0.12,0.25, and 0.06 μg/ml for C. albicans, 0.12, 0.25, and 0.03 μg/ml for C. glabrata complex, 4, 2, and 4 μg/ml for C. parapsilosis complex, 0.5,0.25, and 0.06 μg/ml for C. tropicalis, 0.25,1, and 0.25 μg/ml for C. krusei, 0.25,1, and 0.12 μg/ml for C. lusitaniae, 4,2, and 2 μg/ml for C. guilliermondii, and 0.25,0.25, and 0.12 μg/ml for C. dubliniensis. Species-specific SYO ECVs for anidulafungin, caspofungin, and micafungin correctly classified 72 (88.9%), 74 (91.4%), 76 (93.8%), respectively, of 81 Candida isolates with identified fks mutations. SYO ECVs may aid in detecting non-WT isolates with reduced susceptibility to anidulafungin, micafungin, and especially caspofungin, since testing the susceptibilities of Candida spp. to caspofungin by reference methodologies is not recommended. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

The lack of an ideal antifungal drug or completely successful agent, suggests that combination therapy may be an appropriate option for the management of fungal infections. In this review, different antifungal combination therapy approaches will be discussed. In brief, the rationale for combination therapy is to maximize antifungal effects by attacking different fungal targets at the same time. However, the scientific basis for this approach requires evidence from prospective clinical trials of antifungal combinations. It is possible that real advantages will be seen for particular combinations only in particular mycoses and/or particular types of patient. Combination tends to reduce clinical failure when resistant strains could be recovered from patients, although drug interactions and cross-resistance may result. Synergy has been established between conventional antifungal agents and also between investigational molecules. Alternatively, combination of fluconazole and cyclosporine results fungicidal for fluconazole-susceptible strains of Candida albicans. An overall enhanced susceptibility is reached in intrinsically resistant Candida when combination of azole antifungal drugs or terbinafine with calcineurin inhibitors is used. Azoles, such as voriconazole, itraconazole, and fluconazole are being tested in combination with other antifungal agents such as amphotericin B, caspofungin, or terbinafine and. In addition, amphotericin B is actually administered in combined therapy with caspofungin. The traditional combinations of amphotericin B plus 5-fluorocytosine or amphotericin B plus rifampicin have been replaced by these newer combinations. Other combinations such as caspofungin plus amphotericin B or voriconazole have also been synergic against clinical isolates of Aspergillus and Fusarium. This has been confirmed in animal models in the case of caspofungin plus itraconazole in the treatment of invasive pulmonary aspergillosis and caspofungin plus liposomal amphotericin B in the management of invasive aspergillosis. Fil: Carrillo Muñoz, A. J.. Departamento de Micología; España Fil: Finquelievich, Jorge. Universidad de Buenos Aires. Facultad de Medicina; Argentina Fil: Tur Tur, Cristina. Departamento de Micología; España Fil: Eraso, Elena. Universidad del Pais Vasco; España Fil: Jaureguizar, Nerea. Universidad del Pais Vasco; España Fil: Quindós, Guillermo. Universidad del Pais Vasco; España Fil: Giusiano, Gustavo Emilio. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

To update the knowledge of the epidemiology of fungaemia episodes in Spain, the species implicated and their in vitro antifungal susceptibilities.

Poor outcomes of invasive candidiasis (IC) are associated with the difficulty in establishing the microbiological diagnosis at an early stage. New scores and laboratory tests have been developed in order to make an early therapeutic intervention in an attempt to reduce the high mortality associated with invasive fungal infections. Candida albicans IFA IgG has been recently commercialized for germ tube antibody detection (CAGTA). This test provides a rapid and simple diagnosis of IC (84.4% sensitivity and 94.7% specificity). The aim of this study is to identify the patients who could be benefited by the use of CAGTA test in critical care setting.

The in vitro antifungal activity of a new liposomal nystatin formulation (NISTL, Nyotran, Aronex Ltd., EE.UU.) was evaluated by a microdilution method with RPMI based on the M27A document of the National Committee for Clinical Laboratory Standards (NCCLS) against 22 isolates of Cryptococcus neoformans. This antifungal activity was compared with those of other seven antifungal agents, such as nystatin (NIST), amphotericin B deoxycholate, liposomal amphotericin B, amphotericin B lipid complex, amphotericin B colloidal dispersion, fluconazole, and itraconazole. NISTL was more active in vitrothan NIST, showing MIC values 2-3 fold smaller in 90% of the isolates. The results obtained suggest that this new formulation would be very helpful for the treatment of cryptococcosis.

The utility of Chromalbicans Agar (Biolife Italiana, Milano, Italy) was evaluated with 723 clinical isolates and type culture collection strains from different genera including Candida, Cryptococcus, Pichia, Rhodotorula, Saccharomyces, Trichosporon y Zygosaccharomyces. Presumptive identification was confirmed by germ tube test and carbohydrate assimilation on API-ATB ID 32C (bioMerieux, France). Growth on Chromalbicans Agar was very useful for the presumptive identification of C. albicans isolates, and sensitivity and specificity values were significantly high (97%), since a very low number of isolates were found to be false negative or false positive.

The study constitutes an approach to the knowledge of the epidemiology of cryptococosis in Spain. For detection of cases 167 Spanish hospitals were contacted. All cases included were accompanied by the correspondent isolate of Cryptococcus neoformans, together with clinical, demographic and mycological data. Results obtained from January 1998 to end of December 1999 are analysed and presented here. Fifty-six Spanish hospitals reported 58 cases of cryptococcosis; only 43 of them were adequately documented and accompanied by the clinical isolate. The results showed a higher incidence in males (88.4%) than in females (11.6%); being most frequently affected those between 30 and 40 years old (48.8%). The 84.6% (33) corresponded to new cases and 15.4% (6) to relapses of the disease. The HIV infection was the most frequent risk factor reported (86%) and, for 29.7% (11) of them, cryptococcosis was the AIDS defining disease. For the diagnosis, CSF analysis showed the best results (India ink; culture and antigen detection). All strains collected (100%) corresponded to C. neoformans variety neoformans. Serotypes distribution was 45.5% for serotype A and 22.7% for each of serotypes D and AD.

We studied the possible differences in the pattern of susceptibility to fluconazole and itraconazole in 393 isolates of Candida spp. from the oral cavity of HIV-positive patients and 102 isolates from HIV-negative patients with candidemia or candiduria. We used the broth microdilution method according to the NCCLS guidelines. We observed a decrease in the susceptibility to fluconazole in the group of HIV-positive patients in comparison to those who were HIV negative, especially in Candida albicans (MIC(90) 32 mg/l vs. 1 mg/l and Candida glabrata (MIC(90) 64 mg/l vs. 16 mg/l). Furthermore, we did not find any resistant strains in the HIV-negative group. For itraconazole, the MIC(90) was two dilutions greater in the HIV-positive patients, except for C. albicans, which had a much higher MIC(90) (4 mg/l vs. 0.12 mg/.). Therefore, the decrease in the susceptibility of Candida spp. in the HIV-positive patients must be taken into account when establishing a specific antifungal therapy.

Using Sensititre (AccuMed, USA) we studied the in vitro antifungal activity of amphotericin B, fluconazole, itraconazole, ketoconazole and 5-fluorocytosine against 250 clinical yeast isolates taken from different hospitals, including Candida (151 C. albicans, 15 C. krusei, 14 C. parapsilosis, 11 C. tropicalis, 10 C. glabrata, 4 C. guilliermondii, 3 C. rugosa, 2 C. viswanathii, 2 C. famata and 2 C. kefyr), Cryptococcus (32 C. neoformans and 1 C. laurentii), Trichosporon (2 isolates) and Rhodotorula rubra (1 isolate). All the strains were susceptible to amphotericin B and showed an MIC1 mg/l. The susceptibility of C. albicans (MIC(90)256 mg/l), C. krusei (MIC(90)64 mg/l), C. glabrata (MIC(90)64 mg/l) and C. neoformans (MIC(90) 32 mg/l) to fluconazole was lower (14% isolates being resistant and 16.8% susceptible depending on the dose). The largest number of strains resistant to itraconazole was observed in C. albicans and C. glabrata (17.2% resistant and 24% susceptible and susceptible depending on the dose, respectively). Ketoconazole and 5-fluorocytosine were not effective in vitro against 12.8% and 2%, respectively, of all the isolates studied. Nine C. krusei and seven C. neoformans (12.9%) showed dose-dependent susceptibility to 5-fluorocytosine.

Candida dubliniensis is a recently described Candida species closely related to Candida albicans, which has been associated with oral candidiasis in HIV-infected patients. Fluconazole-resistant strains of C. dubliniensis are easily obtained in vitro and this fact could be a complication if this resistance develops during treatment with this drug.In the present study, the in vitro antifungal susceptibilities of 36 C. dubliniensis clinical isolates and culture strains to current and new antifungal agents, such as amphotericin B (AMB), amphotericin B lipid complex (ABLC), amphotericin B colloidal dispersion (ABCD), 5-fluorocytosine (5FC), fluconazole (FLC), itraconazole (ITC), ketoconazole (KTC), liposomal amphoteri- cin B (LAMB), liposomal nystatin (LNYT), LY303366 (LY), SCH56592 (SCH), and voriconazole (VRC), were determined according to the National Committee for Clinical Laboratory Standards M27-A broth microdilution method for yeasts.Most isolates of C. dubliniensis were susceptible to both new and current antifungal drugs, with 75.9% isolates susceptible to KTC, 86.2% to FLC and to ITC, and approximately 100% to the other antifungal agents tested. The cross-resistance phenotypes are detailed. Four isolates were resistant (MICor =64 microg/ml) to FLC. These 4 isolates were also resistant to KTC, and 3 of them were also resistant to ITC (MICor =1 microg/ml for both agents). However, these isolates were highly susceptible to 5FC and all polyene formulations (AMB, ABLC, ABCD, LAMB, and LNYT), triazole (SCH and VRC) and echinocandin (LY) antifungal agents.The new liposomal and lipidic formulations of AMB, LNYT, and the new triazoles and echinocandins may provide new alternatives to FLC for the treatment of infections by C. dubliniensis.

We report a case of fungemia caused by Candida dubliniensis in a non-HIV infected patient. Multiple cultures of blood performed over a period of 13 days were positive for this recently described yeast species. The C. dubliniensis isolates recovered were susceptible to fluconazole in vitro and the patient responded to intravenous therapy with this antifungal agent. It was possible to differentiate the fungemia caused by C. dubliniensis in this patient from that caused by C. albicans in other patients on the basis of the analysis of the antibody response since the C. dubliniensis-infected patient exhibited a characteristic and specific antibody response against a cell wall component of 160-170 kDa.

The antifungal activity of itraconazole was studied in 101 clinical isolates of Aspergillus fumigatus, A. flavus, A. niger, A. terreus, A. nidulans, A. candidus, A. glaucus, A. clavatus, Fusarium solani, F. oxysporum and F. semitectum. The minimum inhibitory concentrations (MIC) were determined according to the protocol of the M38-P National Committee for Laboratory Standards (NCCLS) document using a microdilution method in 1640 RPMI liquid medium (visual reading at 48 and 72 h incubation). In general, the MIC did not vary with time of incubation, except in a Z. fumigatus strain in which the MIC went from 2 to 16 mg/l. The geometric mean of the MIC and MIC(90) of itraconazole for Aspergillus spp. was 0.44 mg/l and 0.5 mg/l, respectively; and for Fusarium spp. it was 14.1 mg/l and 16 mg/l, respectively. With 0.5 mg/l 75% of the Aspergillus spp. strains were inhibited, and 100% of these strains were inhibited with 2 mg/l. A. niger and A. fumigatus were the most resistant species (MIC(90) 2 mg/l). The MIC of all the Fusarium strains essayed was between 4 and 16 mg/l.