Azole resistance screening in Aspergillus fumigatus sensu stricto using the azole-containing agar method (EUCAST E.Def 10.2): conidial suspension filtration and inoculum adjustment before inoculum preparation may not be needed.

Azole resistance screening in Aspergillus fumigatus sensu stricto can be routinely carried out by using azole-containing agar plates (E.Def 10.2 procedure); however, conidial suspension filtering and inoculum adjustment before inoculum preparation are time-consuming. We evaluated whether skipping the filtration and inoculum adjustment steps negatively influenced the performance of the E.Def 10.2 procedure. A. fumigatus sensu stricto isolates (n = 98), previously classified as azole susceptible or azole resistant (E.Def 9.4 method), were studied. Azole-resistant isolates had either the wild-type cyp51A gene sequence (n = 1) or the following cyp51A gene substitutions: TR34-L98H (n = 41), G54R (n = 5), TR46-Y121F-T289A (n = 1), or G448S (n = 1). In-house azole-containing agar plates were prepared according to the EUCAST E.Def 10.2 procedure. Conidial suspensions obtained by adding distilled water (Tween 20 0.1%) were either filtered and the inocula adjusted to 0.5 McFarland or left unfiltered and unadjusted. Agreements between the agar screening methods using inocula prepared by each procedure were high for itraconazole (99%), voriconazole (100%), and posaconazole (94.9%). Sensitivity and specificity (considering the susceptibility category as per the microdilution E.Def 9.4 method as the gold standard) of E.Def 10.2 were 100% to rule in or rule out resistance when unfiltered and unadjusted suspensions were used; the resistance phenotype of isolates harboring the TR34-L98H, G54R, or TR46-Y121F-T289A substitutions was correctly detected. Unfiltered and unadjusted conidial suspensions do not negatively influence the performance of the E.Def 10.2 method when screening for azole resistance in A. fumigatus sensu stricto. IMPORTANCE: Azole resistance screening in Aspergillus fumigatus sensu stricto can be routinely carried out by using azole-containing plates (E.Def 10.2 procedure); however, conidial suspension filtering and inoculum adjustment before inoculation of plates are time-consuming. We, here, showed that unfiltered and unadjusted conidial suspensions do not negatively influence the performance of the E.Def 10.2 method when screening for azole resistance in A. fumigatus sensu stricto.

Clonal spread of fluconazole-resistant C. parapsilosis in patients admitted to a referral hospital located in Burgos, Spain, during the COVID-19 pandemic.

BACKGROUND: Fluconazole-resistant Candida parapsilosis (FRCP) is a matter of concern in Spain. OBJECTIVES: We here report a FRCP spread across a 777-bed referral hospital located in Burgos, Spain, during the COVID-19 pandemic. PATIENTS/METHODS: In April 2021, an FRCP isolate (MIC = 64 mg/L, E-test®) from a hospitalised patient was detected. Up to June 2022, all C. parapsilosis isolates (n = 35) from hospitalised patients (n = 32) were stored and genotyped using microsatellite markers, and their antifungal susceptibilities were studied (EUCAST); FRCP isolates were molecularly characterised. RESULTS: We detected 26 FRCP isolates collected between 2021 (n = 8) and 2022 (n = 18); isolates were susceptible to amphotericin B, echinocandins and ibrexafungerp. FRCP isolates were grouped into three genotypes: CP-707 and CP-708 involved isolates harbouring the Y132F + R398I ERG11p substitutions (n = 24) and were clonally related; the remaining CP-675 genotype involved isolates harbouring the G458S ERG11p substitution (n = 2). FRCP genotypes were genetically related to the FRCP genotypes found in Madrid and were unrelated to fluconazole-susceptible ones. Patients harbouring FRCP were mainly (n = 22/23) admitted to intensive care units. Most patients had received broad-spectrum antibiotics (n = 22/23), and/or antifungal therapy with azoles (n = 14/23) within the 30 days prior to FRCP isolation. Thirteen patients were colonised, 10 of whom were infected and presented candidaemia (n = 8/10), endovascular infection (n = 1/10) or complicated urinary infection (n = 1/10). Overall nonattributable 30-day mortality was 17% (n = 4/23). CONCLUSIONS: We report an outbreak caused by FRCP affecting patients admitted to the ICU of a referral hospital located in Burgos. Patients harbouring FRCP had a higher fluconazole use than those carrying susceptible isolates.

Fluconazole-resistant Candida parapsilosis genotypes from hospitals located in five Spanish cities and one in Italy: Description of azole-resistance profiles associated with the Y132F ERG11p substitution.

BACKGROUND: Fluconazole-resistant Candida parapsilosis is a matter of concern. OBJECTIVES: To describe fluconazole-resistant C. parapsilosis genotypes circulating across hospitals in Spain and Rome and to study their azole-resistance profile associated with ERG11p substitutions. PATIENTS/METHODS: We selected fluconazole-resistant C. parapsilosis isolates (n = 528 from 2019 to 2023; MIC ≥8 mg/L according to EUCAST) from patients admitted to 13 hospitals located in five Spanish cities and Rome. Additionally, we tested voriconazole, posaconazole, isavuconazole, amphotericin B, micafungin, anidulafungin and ibrexafungerp susceptibility. RESULTS: Of the 53 genotypes found, 49 harboured the Y132F substitution, five of which were dominating city-specific genotypes involving almost half the isolates. Another genotype involved isolates harbouring the G458S substitution. Finally, we found two genotypes with the wild-type ERG11 gene sequence and one with the R398I substitution. All isolates were fully susceptible/wild-type to amphotericin B, anidulafungin, micafungin and ibrexafungerp. The azole-resistance patterns found were: voriconazole-resistant (74.1%) or voriconazole-intermediate (25.2%), posaconazole-resistant (10%) and isavuconazole non-wild-type (47.5%). Fluconazole-resistant and voriconazole non-wild-type isolates were likely to harbour substitution Y132F if posaconazole was wild type; however, if posaconazole was non-wild type, substitution G458S was indicated if isavuconazole MIC was >0.125 mg/L or substitution Y132F if isavuconazole MIC was ≤0.125 mg/L. CONCLUSIONS: We detected a recent clonal spread of fluconazole-resistant C. parapsilosis across some cities in Spain, mostly driven by dominating city-specific genotypes, which involved a large number of isolates harbouring the Y132F ERG11p substitution. Isolates harbouring substitution Y132F can be suspected because they are non-susceptible to voriconazole and rarely posaconazole-resistant.

Twenty Years in EUCAST Anti-Fungal Susceptibility Testing: Progress & Remaining Challenges.

Since its inception in 2002, the EUCAST Antifungal Susceptibility Testing Subcommittee (AFST) has developed and refined susceptibility testing methods for yeast, moulds and dermatophytes, and established epidemiological cut-off values and breakpoints for antifungals. For yeast, three challenges have been addressed. Interpretation of trailing growth in fluconazole susceptibility testing, which has been proven without impact on efficacy if below the 50% endpoint. Variability in rezafungin MIC testing due to laboratory conditions, which has been solved by the addition of Tween 20 to the growth medium in E.Def 7.4. And third, interpretation of MICs for rare yeast with no breakpoints, where recommendations have been established for MIC-based clinical advice. For moulds, refinements include the validation of spectrophotometer reading for A. fumigatus to facilitate objective MIC determination, and for dermatophytes the establishment of a microdilution method with automated reading and a selective medium to minimise the risk of contaminations. Recent initiatives involve development and validation of agar-based screening assays for detection of potential azole and echinocandin resistance in A. fumigatus and Aspergillus species, respectively, and of terbinafine resistance in Trichophyton species. Moreover, the development of a EUCAST guidance document for molecular resistance testing represents an advancement, particularly for identifying target gene alterations associated with resistance. In summary, EUCAST AFST continues to play a pivotal role in standardizing AFST and facilitating accurate interpretation of susceptibility data for clinical decision-making. Adoption of EUCAST breakpoints for commercial test methods, however, requires thorough validation to ensure concordance with EUCAST reference testing species-specific MIC distributions.

Aspergillus granulosus femoral osteomyelitis in a cardiac transplant patient: first reported case and literature review.

Aspergillus osteomyelitis is a rare complication of extrapulmonary invasive aspergillosis, which usually presents as spondylodiscitis. The clinical picture is usually paucisymptomatic and of long evolution, which leads to diagnostic difficulties, especially in immunosuppressed patients presenting a delayed systemic host response. We report a case of femoral osteomyelitis caused by Aspergillus granulosus in a heart transplant recipient successfully treated with a combined surgical and antifungal approach. A 65-year-old heart transplant male presented with left knee pain lasting 3 months. X-ray and magnetic resonance imaging identified a lesion with aggressive characteristics at the distal third of the left femur, due to which the patient underwent excisional surgery. Aspergillus granulosus was cultured from the removed material and antifungal treatment with oral isavuconazole was started. Chest imaging excluded pulmonary aspergillosis, while the positron emission tomography/computed tomography (PET/CT) identified a remnant of a prosthetic vascular graft sewn to the proximal third of the right axillary artery, through which a catheter-based micro-axial left ventricular assist device was implanted previously as bridge to transplant therapy. The patient presented a rapid clinical improvement with complete functional recovery following the surgical treatment and the antifungal therapy and finally underwent surgical removal of the residual vascular graft. This is the first reported episode of long bone osteomyelitis due to A. granulosus that occurred in a heart transplant recipient without pulmonary infection and was successfully treated with isavuconazole. The PET/CT was useful in supporting the diagnostic process and follow-up. Cryptic fungal species can cause invasive infections, particularly in immunocompromised patients. Molecular methods are crucial in fungal identification.

Fluconazole-resistant Candida parapsilosis: fast detection of the Y132F ERG11p substitution, and a proposed microsatellite genotyping scheme.

OBJECTIVES: We propose fast and accurate molecular detection of the Y132F ERG11p substitution directly on pure-cultured Candida parapsilosis isolates. We also assessed a discriminative genotyping scheme to track circulating genotypes. METHODS: A total of 223 C. parapsilosis isolates (one patient each) from 20 hospitals, located in Spain and Italy were selected. Isolates were fluconazole-resistant (n = 94; harbouring the Y132F ERG11p substitution [n = 85], the G458S substitution [n = 6], the R398I substitution [n = 2], or the wild-type ERG11 gene sequence) or fluconazole-susceptible (n = 129). Two targeted-A395T-mutation PCR formats (conventional and real-time) were engineered and optimized on fluconazole-susceptible and fluconazole-resistant pure-cultured isolates, thus skipping DNA extraction. Two genotyping schemes were compared: Scheme 1 (CP1, CP4a, CP6, and B markers), and Scheme 2 (6A, 6B, 6C, CP1, CP4a, and CP6 markers). RESULTS: The screening performed using both PCR formats showed 100% specificity (fluconazole-susceptible isolates; n = 129/129) and sensitivity (Y132F isolates; n = 85/85) values; however, results were available in 3 and 1.5 hours with the conventional and real-time PCR formats, respectively. Overall, Scheme 1 showed higher genetic diversity than Scheme 2, as shown by the number of alleles detected (n = 98; mean 23, range 13-38), the significantly higher observed and expected heterozygosity, and the probability of identity index (2.5 × 10-6). Scheme 2 markers did not provide further genotypic discrimination of Y132F fluconazole-resistant genotypes. CONCLUSION: Both proposed PCR formats allow us to speed up the accurate detection of substitution Y132F ERG11p in C. parapsilosis isolates with 100% specificity and sensitivity. In addition, we recommend CP1, CP4a, CP6, and B microsatellite markers for genotyping fluconazole-resistant isolates.

Air pollution and health prevention: A document of reflection / Calidad del aire ambiente y prevención de la salud: Un documento de reflexión

Ambient air quality, pollution and its implication onhealth is a topic of enormous importance that is normallydealt with by major specialists in their particular areas of interest. In general, it is not discussed from multidisciplinary approaches or with a language that can reach everyone. For thisreason, the Health Sciences Foundation, from its preventionarea, has formulated a series of questions to people with veryvaried competences in the area of ambient air quality in orderto obtain a global panorama of the problem and its elementsof measurement and control. The answers have been produced by specialists in each subject and have been subjected to ageneral discussion that has allowed conclusions to be reached on each point. The subject was divided into three main blocks:external ambient air, internal ambient air, mainly in the workplace, and hospital ambient air and the consequences of itspoor control. Along with the definitions of each area and theindicators of good and bad quality, some necessary solutionshave been pointed out. We have tried to know the current legislation on this problem and the competences of the differentadministrations on it. Despite its enormous importance, ambient air quality and health is not usually a topic of frequentpresence in the general media and we have asked about thecauses of this. Finally, the paper addresses a series of reflections from the perspective of ethics and very particularly in thelight of the events that the present pandemic raises. This workaims to provide objective data and opinions that will enablenon-specialists in the field to gain a better understanding ofthis worrying reality. (AU)

La calidad del aire ambiente y su implicación en la salud esun tema de enorme importancia que normalmente es tratadopor grandes especialistas en sus particulares áreas de interés.En general, no es discutido desde enfoques multidisciplinaresni con un lenguaje que pueda llegar a todos. Por ese motivo, laFundación de Ciencias de la Salud desde su área de prevención,ha formulado una serie de preguntas a personas con competencias muy variadas en el área de la calidad del aire ambientepara obtener un panorama global del problema y de sus elementos de medida y control. Las respuestas han sido producidas por especialistas en cada tema y han sido sometidas a unadiscusión general que ha permitido alcanzar conclusiones encada punto. El tema ha sido dividido en tres grandes bloques:el aire ambiente externo, el aire ambiente interno, principalmente en el medio laboral, y el aire ambiente hospitalario ylas consecuencias de su mal control. Junto con las definicionesde cada área y los indicadores de buena y mala calidad, se haapuntado a algunas necesarias soluciones. Hemos tratado deconocer la legislación vigente sobre este problema y las competencias de las distintas administraciones sobre el mismo.Pese a su enorme importancia, la calidad del aire ambiente yla salud no suele ser un tema de frecuente presencia en losmedios de comunicación generales y hemos preguntado sobrelas causas de ello. Finalmente, el documento aborda una seriede reflexiones desde la perspectiva de la ética y muy particularmente a la luz de los acontecimientos que la presente pandemia plantea. Este trabajo pretende aportar datos objetivos yopinión que permitan a los no especialistas en el tema conocermejor esta preocupante realidad. (AU)

Del CLSI al EUCAST, una transición necesaria en los laboratorios españoles / From CLSI to EUCAST, a necessary step in Spanish laboratories

El Comité Español del Antibiograma (COESANT) presenta en este documento una sencilla «hoja de ruta» en forma de decálogo de recomendaciones cuya finalidad es facilitar la transición de la normativa del Clinical and Laboratory Standards Institute (CLSI) a la del European Committee on Antimirobial Susceptibility Testing (EUCAST) en los Servicios y Unidades de Microbiología Clínica que aún emplean los criterios del CLSI. Su objetivo es adaptar las directrices europeas, más próximas a la realidad clínico-epidemiológica española, y conseguir una implantación de los criterios del EUCAST en la totalidad de los laboratorios de Microbiología en España

The Spanish Antibiogram Committee (Comité Español del Antibiograma, COESANT) presents in this document a simple "roadmap" or decalogue of recommendations, with a view to facilitating the transition from the Clinical and Laboratory Standards Institute (CLSI) to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) antimicrobial susceptibility testing regulations to the Clinical Microbiology Spanish laboratories that still use the CLSI guidelines. The objectives are to adapt the closer European regulations to the Spanish clinical and epidemiological reality and to fully implement the EUCAST recommendations in all microbiology laboratories in Spain

Recommendations of the Spanish Antibiogram Committee (COESANT) for selecting antimicrobial agents and concentrations for in vitro susceptibility studies using automated systems / Recomendaciones del Comité Español del Antibiograma (COESANT) para la selección de antimicrobianos y sus concentraciones en el estudio in vitro de la sensibilidad con métodos automáticos

Automated antimicrobial susceptibility testing devices are widely implemented in clinical microbiology laboratories in Spain, mainly using EUCAST (European Committee on Antimicrobial Susceptibility Testing) breakpoints. In 2007, a group of experts published recommendations for including antimicrobial agents and selecting concentrations in these systems. Under the patronage of the Spanish Antibiogram Committee (Comité Español del Antibiograma, COESANT) and the Study Group on Mechanisms of Action and Resistance to Antimicrobial Agents (GEMARA) from the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC), and aligned with the Spanish National Plan against Antimicrobial Resistance (PRAN), a group of experts have updated this document. The main modifications from the previous version comprise the inclusion of new antimicrobial agents, adaptation of the ranges of concentrations to cover the EUCAST breakpoints and epidemiological cut-off values (ECOFFs), and the inference of new resistance mechanisms. This proposal should be considered by different manufacturers and users when designing new panels or cards. In addition, recommendations for selective reporting are also included. With this approach, the implementation of EUCAST breakpoints will be easier, increasing the quality of antimicrobial susceptibility testing data and their microbiological interpretation. It will also benefit epidemiological surveillance studies as well as the clinical use of antimicrobials aligned with antimicrobial stewardship programs

Los sistemas automáticos utilizados en el estudio de la sensibilidad a los antimicrobianos están introducidos en la mayoría de los laboratorios de Microbiología Clínica en España, utilizando principalmente los puntos de corte del European Committee on Antimicrobial Susceptibility Testing (EUCAST). En 2007, un grupo de expertos publicó unas recomendaciones para incluir antimicrobianos y seleccionar concentraciones en estos sistemas. Bajo el auspicio del Comité Español del Antibiograma (COESANT) y del Grupo de Estudio de los Mecanismos de Acción y Resistencia a los Antimicrobianos (GEMARA) de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC) y alineado con el Plan Nacional frente a la Resistencia a los Antibióticos (PRAN), un grupo de expertos ha actualizado dicho documento. Las principales modificaciones realizadas sobre la versión anterior comprenden la inclusión de nuevos agentes antimicrobianos, la adaptación de los rangos de concentraciones para cubrir los puntos de corte clínicos y los puntos de corte epidemiológicos (ECOFF) definidos por el EUCAST, y para la inferencia de nuevos mecanismos de resistencia. Esta propuesta debería ser considerada por los diferentes fabricantes y los usuarios cuando se diseñen nuevos paneles o tarjetas. Además, se incluyen recomendaciones para realizar informes selectivos. Con este enfoque, la implementación de los puntos de corte del EUCAST será más fácil, aumentando la calidad de los datos del antibiograma y su interpretación microbiológica. También será de utilidad para los estudios de vigilancia epidemiológica, así como para el uso clínico de los antimicrobianos, de acuerdo con los programas de optimización de uso de antimicrobianos (PROA)

Infectious endocarditis caused by Candida glabrata: evidence of in vivo development of echinocandin resistance / Endocarditis infecciosa por Candida glabrata: evidencia del desarrollo in vivo de resistencia a equinocandinas

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Recomendaciones GEMICOMED/GEIRAS-SEIMC para el manejo de las infecciones y colonizaciones por Candida auris / GEMICOMED/GEIRAS-SEIMC recommendations for the management of Candida auris infection and colonization

Candida auris es una nueva especie de Candida responsable de diversos brotes nosocomiales en varios países del mundo, incluida España; es resistente al fluconazol y se han descrito cepas multi y panresistentes. Presenta una elevada transmisibilidad y extensa supervivencia en el entorno hospitalario, lo que es causa de brotes de larga duración difíciles de detectar en fases tempranas y dificulta su control e intento de erradicación. C.auris constituye actualmente una amenaza emergente para la salud global. Para limitar el impacto y facilitar el control de la infección por C.auris, el presente documento ofrece un conjunto de recomendaciones basadas en las experiencias obtenidas en los brotes de España y del Reino Unido, elaboradas por un equipo multidisciplinar. La puesta en marcha de medidas de vigilancia y control es esencial para evitar la propagación del brote, que puede prolongarse en el tiempo y representar así un riesgo importante para los pacientes quirúrgicos complejos, críticos e inmunocomprometidos. La notificación inmediata del aislamiento de C.auris a los equipos clínicos y de control de infecciones, así como a las autoridades e instituciones sanitarias, es esencial para implementar las medidas de control de infecciones a todos los niveles y escalas de manera oportuna, para evitar la transmisión interna e intercentros, y para garantizar la vigilancia y la prevención del desarrollo de infecciones en pacientes que ya se encuentran colonizados

Candida auris is a new species of Candida that causes nosocomial outbreaks in several countries around the world, including Spain. C.auris is resistant to fluconazole and multi- and pan-resistant strains have been described. It is highly transmissible and can survive long term in the hospital environment, causing long-lasting outbreaks that are difficult to detect in early stages, and making it difficult to control and eradicate. It is currently an emerging threat to global health. This document provides a set of guidelines, developed by a multidisciplinary team, to limit the impact and facilitate the control of C.auris infection based on the experiences gathered in the Spanish and English outbreaks. The implementation of early and strict surveillance and control measures is essential to prevent the spread of the outbreak, which can spread over time, posing a significant risk to complex, critical and immunocompromised surgical patients. Immediate notification of C.auris isolation to clinical and infection control teams, as well as to health authorities and institutions, is essential to implement infection control measures at all levels in a timely manner, to prevent internal and inter-centre transmission, and to ensure a proper surveillance and prevention to patients who are already colonized and can develop an infection

Executive summary of clinical practice guideline for the management of invasive diseases caused by Aspergillus: 2018 Update by the GEMICOMED-SEIMC/REIPI / Resumen ejecutivo del documento de consenso del GEMICOMED perteneciente a la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC) sobre el tratamiento de las infecciones invasoras producidas por Aspergillus

Aspergillus infection is a significant cause of morbi-mortality in an at-risk population. The Study Group of Fungal Infections (GEMICOMED) from the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC) has reviewed announcements made in invasive aspergillosis management. We have organized our recommendations in such a way as to provide a guide in resolving different clinical situations concerning the entire spectrum of invasive diseases caused by Aspergillus in various populations. Diagnostic approach, treatment and preventions strategies are outlined. It is not our aim that these guidelines supplant clinical judgment with respect to specific patients; however, it is our objective to perform a comprehensive summary of quality of care evidence for invasive aspergillosis management in different settings

Las infecciones causadas por Aspergillus causan una elevada morbimortalidad en la población susceptible. EL Grupo de Estudio de Micología Médica de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (GEMICOMED/SEIMC) ha revisado las novedades más importantes sobre el manejo de las infecciones invasoras causadas por Aspergillus. Hemos organizado nuestras recomendaciones en 3 apartados: diagnóstico, tratamiento y profilaxis en diferentes grupos de pacientes susceptibles de padecer estas infecciones. Se revisan distintas situaciones clínicas que pueden estar causadas por este hongo. Nuestro objetivo no es que estas guías de tratamiento suplanten el juicio clínico de los médicos ante un determinado paciente; sin embargo, sí deseamos poder ofrecer un resumen comprensible sobre las evidencias que existen para realizar un óptimo manejo de la infección invasora causada por Aspergillus en diferentes situaciones clínicas

Diagnóstico microbiológico de las infecciones relacionadas con la formación de biopelículas / Microbiological diagnosis of biofilm-related infections

Las infecciones asociadas a biopelículas suponen un grave problema sanitario ya que representan entre el 65 y el 80% de todas las infecciones. Estas son generalmente crónicas y están caracterizadas por la persistencia del microorganismo debido a su resistencia al sistema inmunitario y a los antimicrobianos. Las biopelículas se pueden localizar tanto en tejidos humanos como sobre dispositivos exógenos tales como catéteres, marcapasos, prótesis, implantes, sondas urinarias, etc. Tradicionalmente, los laboratorios de microbiología clínica realizan los estudios de sensibilidad sobre microorganismos en crecimiento planctónico. Sin embargo, de esta manera se pierden las características propias de la biopelícula con lo que la antibioterapia basada en estos estudios podría asociarse con fracaso terapéutico o recurrencias. El diagnóstico microbiológico y los estudios de sensibilidad en las infecciones relacionadas con biopelículas son complejos y, hoy por hoy, representan un reto que clínicos y microbiólogos han de abordar en equipo ya que no existe todavía un consenso global ni protocolos estandarizados

Biofilm-related infections represent a serious health problem, accounting for 65- 80% of all infections. The infections are generally chronic and characterized by the persistence of the microorganism, due to the increased resistance of biofilms to both the immune system and antimicrobials. Biofilms can be located to almost every human body tissue and on exogenous devices such as catheters, pacemakers, prosthetic material, implants, urinary catheters, etc. Traditional antimicrobial susceptibility studies in clinical microbiology laboratories have lied on the study of planktonic form of microorganisms. However, this approach might lead to miss the biofilm characteristics and to a treatment failure. Microbiological diagnosis and antimicrobial susceptibility studies of biofilm-related infections are complex and, nowadays, represent a challenge that clinicians and microbiologists have to address as a team in the absence of consensus or standardized protocols

Recommendations of the Spanish Antibiogram Committee (COESANT) for in vitro susceptibility testing of antimicrobial agents by disk diffusion / Recomendaciones del Comité Español del Antibiograma (COESANT) para el estudio de sensibilidad in vitro a los antimicrobianos por difusión con discos

Disk diffusion is a well standardized method that provides reliable categorical results to guide antimicrobial therapy in numerous types of infections. Based on the guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST), which are widely implemented in Spain, the Spanish Antibiogram Committee (COESANT) has drawn up recommendations for antimicrobial selection by the disk diffusion technique, including selective reporting and its use for the detection of resistance mechanisms. Factors affecting disk diffusion results, along with advantages and shortcomings of the method, are also discussed.(AU)

La difusión con discos es un método estandarizado que proporciona resultados fiables para guiar la terapia antimicrobiana en numerosos tipos de infecciones. En base a las directrices del European Committee on Antimicrobial Susceptibility Testing (EUCAST), ampliamente implantadas en España, el Comité Español del Antibiograma (COESANT) ha elaborado recomendaciones para la selección de antimicrobianos para ser estudiados mediante la técnica de difusión con discos, su notificación selectiva en el informe de sensibilidad y su uso para la detección de mecanismos de resistencia. También se discuten los factores que afectan los resultados obtenidos mediante la técnica de difusión con discos junto con las ventajas y desventajas del método.(AU)

Recomendaciones del Comité Español del Antibiograma (COESANT) para la realización de los Informes de Sensibilidad Antibiótica Acumulada / Spanish Antibiogram Committee (COESANT) recommendations for cumulative antibiogram reports

El Comité Español del Antibiograma (COESANT) presenta en este documento una serie de recomendaciones cuya finalidad es unificar la forma en la que los Servicios y Unidades de Microbiología Clínica españoles realizan los informes de sensibilidad acumulada de las bacterias, aisladas en muestras clínicas, frente a los antimicrobianos. Las recomendaciones se fundamentan en las recogidas en el Procedimiento de Microbiología Clínica n° 51, «Preparación de informes acumulados de sensibilidad a los antimicrobianos» de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC), publicado en 2014, y recoge las modificaciones en las definiciones de las interpretaciones de las categorías clínicas publicadas en el año 2019 por el European Committee on Antimicrobial Susceptibility Testing (EUCAST). Su objetivo final es establecer una forma homogénea de elaborar estos resúmenes para poder comparar resultados de diferentes centros o sumar su información y así realizar una adecuada vigilancia local o incluso nacional de la evolución de la sensibilidad a los antimicrobianos.(AU)

The Spanish Antibiogram Committee (Comité Español del Antibiograma, COESANT) presents in this document a series of recommendations intending to unify how cumulative antibiogram reports must be made in Clinical Microbiology Spanish laboratories. This article is based on the information included in the Clinical Microbiology Procedure No. 51, «Preparation of cumulative reports on antimicrobial susceptibility» of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC), published in 2014. The recommendations also include the modifications in the definition of clinical interpretive categories recently published by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) in 2019. Its final objective is to establish a homogeneous way of preparing these summaries to compare results from different centers or aggregate the information from these in order to carry out an adequate local or even national surveillance regarding the evolution of antimicrobial susceptibility.(AU)

Fluconazole-containing agar Sabouraud dextrose plates are not useful when screening for susceptibility in Candida albicans / Las placas de agar dextrosado de Sabouraud con fluconazol no son útiles para la detección de resistencia en Candida albicans

Introduction. Fluconazole is an alternative for candidemic patients who are not critically ill. Fluconazole is mainly fungistatic and does not completely inhibit visual Candida albicans growth. We studied the usefulness of fluconazolecontaining Sabouraud dextrose agar plates for detecting susceptibility to fluconazole in C. albicans. Material and methods. Adjusted inocula of 19 isolates were transferred directly onto fluconazole-containing Sabouraud dextrose plates (concentrations ranging from 0.125 mg/L to 128 mg/L). The fluconazole MIC in fresh isolates and after growth on the fluconazole-containing plate at 128 mg/L was recorded following the EUCAST EDef 7.2 guidelines. Then isolates were classified according to their degree of trailing production, based on microdilution procedure. Results. All isolates were able to grow on all fluconazolecontaining plates, even those isolates susceptible to fluconazole. In fact, we selected isolates with different degrees of trailing based on microdilution procedures. 50% of isolates classified as heavy trailers, 35.71% as moderate trailers, and 14.28% as slight trailers. Conclusions. The use of fluconazole-containing Sabouraud dextrose agar plates was not a reliable method to detect fluconazole susceptibility in C. albicans isolates; growth of the isolates was a trailing effect rather than true resistance (AU)

Introducción. Fluconazol es el antifúngico de elección en pacientes con candidemia que no están en estado crítico. Fluconazol es un fármaco fungistático y no inhibe por completo el crecimiento de Candida albicans. En este estudio se evalúa la posibilidad de emplear placas de agar dextrosado de Sabouraud con diferentes concentraciones de fluconazol como método para evaluar la sensibilidad a este fármaco de aislados de C. albicans. Material y métodos. El inóculo ajustado procedente de 19 aislados de C. albicans se transfirió directamente a placas que contenían concentraciones de fluconazol comprendidas entre 0,125 mg/L y 128 mg/L. La CMI de fluconazol se calculó en los aislados originales y en los aislados crecidos en la placa de fluconazol de 128 mg/L, según el protocolo de EUCAST EDef 7.2. Los aislados se clasificaron según su grado de producción de efecto de arrastre, siguiendo el procedimiento de microdilución. Resultados. Todos los aislados fueron capaces de crecer en todas las placas con diferentes concentraciones de fluconazol, incluso las cepas sensibles. Los aislados mostraron diferentes grados de efecto de arrastre. El 50% de los aislados se clasificaron como altamente productores, el 35,71% como moderadamente productores y el 14,28% como poco productores de efecto de arrastre. Conclusión. El uso de placas de agar dextrosado de Sabouraud con fluconazol no demostró ser un método útil para el estudio de la sensibilidad de C. albicans a este fármaco ya que el crecimiento de los aislados fue interpretado como efecto de arrastre y no como una verdadera resistencia (AU)

Abordaje de la candidemia y la candidiasis invasiva en el paciente crítico: papel de las equinocandinas / Candidemia and invasive candidiasis approach in critically ill patients: role of the echinocandins

Introducción. Las infecciones invasoras producidas por Candida spp. en pacientes críticos pueden empeorar considerablemente su pronóstico, por lo que es de gran importancia establecer una detección precoz y una estrategia terapéutica adecuada. El objetivo de este estudio ha sido definir el papel diferencial de las equinocandinas para tratar determinados perfiles de pacientes críticos. Metodología. Un comité científico formado por 9 expertos en enfermedades infecciosas, cuidados críticos, microbiología y farmacia hospitalaria revisó la evidencia existente sobre el tratamiento de la candidemia y la candidiasis invasiva en pacientes críticos. Tras ello, se elaboró un cuestionario con 35 aseveraciones para ser consensuadas por 26 especialistas de las disciplinas mencionadas mediante un método Delphi modificado. Resultados. Después de dos rondas de evaluación, se alcanzó un consenso de acuerdo en el 66% de las aseveraciones. Entre los acuerdos alcanzados son: no es necesario ajustar la dosis de equinocandinas durante una terapia de reemplazo renal; las equinocandinas son el tratamiento empírico y/o dirigido de elección para la candidemia y la candidiasis invasiva asociada a biopelículas; estos fármacos pueden utilizarse en la profilaxis antifúngica del trasplante hepático de alto riesgo. En ausencia de datos clínicos adicionales, se ha de señalar que micafungina es la equinocandina con mayor evidencia científica. Conclusiones. Los expertos consultados mostraron un alto grado de acuerdo sobre algunos de los aspectos más controvertidos relativos al manejo de la candidemia y la candidiasis invasiva en pacientes críticos, lo permitiría aportar recomendaciones prácticas para su tratamiento (AU)

Introduction. Invasive infections caused by Candida spp. in critically ill patients may significantly worsen their prognosis, so it is of great importance to establish an early detection and a suitable therapeutic strategy. The objective of this study was to define the differential role of echinocandins in treating certain critical patient profiles. Methodology. A scientific committee of 9 experts in infectious diseases, critical care, microbiology, and hospital pharmacy reviewed the existing evidence on the treatment of candidemia and invasive candidiasis in critically ill patients. After that, a questionnaire with 35 items was elaborated to be agreed by 26 specialists in the aforementioned disciplines using a modified Delphi method. Results. After two rounds of evaluation, a consensus was reached in terms of agreement in 66% of the items. Some of the consensuses achieved included: it is not necessary to adjust the dose of echinocandins during renal replacement therapy; the echinocandins are the empirical and/or directed treatment of choice for candidemia and invasive candidiasis associated with biofilms; these drugs may be used in the antifungal prophylaxis of high-risk liver transplantation. In the absence of additional clinical data, it should be noted that micafungin is the echinocandin with the most available scientific evidence. Conclusions. The experts consulted showed a high degree of agreement on some of the most controversial aspects regarding the management of candidemia and invasive candidiasis in critical patients, which could inform of practical recommendations for their treatment (AU)

Lack of relationship between genotype and virulence in Candida species / Ausencia de relación entre el genotipo y la virulencia en especies de Candida

BACKGROUND: The virulence of isolates among different Candida species causing candidemia may play a role in the prognosis of the patients. Furthermore, the potential relationship between genotype and virulence is still unclear and need to be further studied. AIMS: We aim to assess the relationship between genotype and virulence in Candida species using a Galleria mellonella larvae infection model. METHODS: One hundred and ninety-four isolates from 68 clusters (Candida albicans, 114/41; Candida parapsilosis, 74/24; Candida tropicalis, 6/3) were compared against the same number of each species singleton genotypes in terms of survival of G. mellonella larvae. RESULTS: The median of survival and the IQR ranges of clusters and singleton were as follows: C. albicans (2 days, IQR 1.5-2 vs. 2 days, IQR 1-2.25), C. parapsilosis (2 days, IQR 1.5-2.6 vs. 2 days, IQR 2-3.3), and C. tropicalis (1 day, IQR 1-3.5 vs. 2 days, IQR 2-3.5; p < 0.05). High intra-cluster variability in terms of median of survival was found regardless the species. CONCLUSIONS: No relationship between genotype and virulence in Candida was observed with the G. mellonella model

ANTECEDENTES: La virulencia de cepas de diferentes especies de Candida causantes de candidemia puede jugar un papel en el pronóstico de los pacientes, y su estudio en el modelo de infección en Galleria mellonella puede ser útil para entender su contribución general a la infección. Además, la potencial relación entre genotipo y virulencia requiere de más estudios. OBJETIVOS: Se evaluó la relación entre genotipo y virulencia en especies de Candida mediante el modelo de infección de larvas de G. mellonella. MÉTODOS: Se estudió la supervivencia de las larvas infectadas con 194 aislados incluidos en 68 clusters (Candida albicans, 114/41; Candida parapsilosis, 74/24; Candida tropicalis, 6/3) y con el mismo número de aislados con genotipos únicos por especie. RESULTADOS: La mediana de supervivencia y los rangos intercuartílicos (IQR) de clusters y genotipos únicos se muestra a continuación: C. albicans (2 días, IQR: 1,5-2 vs. 2 días, IQR: 1-2,25), C. parapsilosis (2 días, IQR: 1,5-2,6 vs. 2 días, IQR: 2-3,3), y C. tropicalis (un día, IQR: 1-3,5 vs. 2 días, IQR: 2-3,5; p < 0,05). Encontramos una importante variabilidad en la mediana de supervivencia entre cepas del mismo cluster, independientemente de la especie analizada. CONCLUSIONES: No se encontró relación entre el genotipo y la virulencia entre los aislados de Candida evaluados mediante el modelo de infección de G. mellonella

Training should be the first step toward an antifungal stewardship program

The frequency of use of systemic antifungal agents has increased significantly in most tertiary centers. However, antifungal stewardship has received very little attention. The objective of this article was to assess the knowledge of prescribing physicians in our institution as a first step in the development of an antifungal stewardship program. Attending physicians from the departments that prescribe most antifungals were invited to complete a questionnaire based on current guidelines on diagnosis and therapy of invasive candidiasis and invasive aspergillosis (IA).The survey was completed by 60.8% (200/329) of the physicians who were invited to participate. The physicians belonged to the following departments: medical (60%), pediatric (19%), intensive care (15.5%), and surgical (5.5%). The mean (±SD) score of correct responses was 5.16 ± 1.73. In the case of candidiasis, only 55% of the physicians clearly distinguished between colonization and infection, and 17.5% knew the local rate of fluconazole resistance. Thirty-three percent knew the accepted indications for antifungal prophylaxis, and 23% the indications for empirical therapy. However, most physicians knew which antifungals to choose when starting empirical therapy (73.5%). As for aspergillosis, most physicians (67%) could differentiate between colonization and infection, and 34.5% knew the diagnostic value of galactomannan. The radiological features of IA were well recognized by 64%, but only 31.5% were aware of the first line of treatment for IA, and 36% of the recommended duration of therapy. The usefulness of antifungal levels was known by 67%. This simple, easily completed questionnaire enabled us to identify which areas of our training strategy could be improved

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Aplicación de las técnicas de tipificación molecular al estudio de brotes hospitalarios de candidemia / Use of molecular typing tools for the study of hospital outbreaks of candidemia

La candidemia es una complicación infecciosa que afecta fundamentalmente a pacientes ingresados en unidades de cuidados intensivos así como en otros servicios hospitalarios, y cuya mortalidad puede alcanzar el 40%. La candidemia es una infección de adquisición típicamente hospitalaria, por lo que la transmisión horizontal de Candida spp. puede traducirse en la aparición de brotes nosocomiales. La caracterización genotípica de los aislamientos de Candida causantes de candidemia puede ayudar a esclarecer el origen de la infección, detectar los servicios hospitalarios con transmisión activa y, consecuentemente, mejorar su prevención. Diversas técnicas de tipificación molecular han sido empleadas para el estudio genotípico de aislamientos de Candida. Las técnicas basadas en microsatélites son reproducibles y presentan un alto poder de discriminación, lo que las convierte en opciones atractivas para el estudio de brotes de candidemia. La mayor parte de los brotes han sido descritos en pacientes ingresados en unidades de cuidados intensivos, fundamentalmente en neonatos. La presente revisión pretende discutir el papel de la caracterización genotípica de aislamientos de Candida causantes de candidemia para el estudio de brotes hospitalarios, así como describir las poblaciones más frecuentemente afectadas por las cepas epidémicas (AU)

Candidemia is an infectious complication mainly affecting hospitalized patients, particularly those admitted to intensive care units. Patient mortality can reach up to 40%. Candidemia is typically nosocomially-acquired, and horizontal transmission of Candida spp. can lead to the presence of outbreaks of candidemia. Genotyping of isolates of Candida causing candidemia can help us to understand the source of the infection, detect the hospital wards with active Candida spp. transmission and, consequently, improve the prevention of the infection. Several genotyping tools have been used for the molecular characterization of Candida isolates involved in outbreaks of candidemia. Genotyping procedures based on microsatellites are reproducible and show a high discriminatory power. Microsatellites are recommended for the study of outbreaks of candidemia. In most hospital outbreaks of candidemia, patients admitted to intensive care units are involved, mostly neonatal patients. The role of genotyping Candida isolates causing candidemia for the study of nosocomial outbreaks of candidemia is reviewed, as well as the patients more commonly affected by epidemic strains (AU)