Antifungal susceptibility testing following the CLSI M27 document, along with the measurement of MFC/MIC ratio, could be the optimal approach to detect amphotericin B resistance in Clavispora (Candida) lusitaniae. Susceptibility patterns of contemporary isolates of this species.

Antifungal susceptibility testing (AST) is crucial in clinical settings to guide appropriate therapy. Nevertheless, discrepancies between treatment response and some results still persist, particularly in detecting resistance to amphotericin B (AMB) in Clavispora (Candida) lusitaniae. This study aimed to assess the susceptibility patterns of 48 recent isolates of C. lusitaniae to 9 antifungal agents and explore the feasibility of using a CLSI reference-based method to identify AMB resistance. Microdilution techniques revealed a wide range of minimal inhibitory concentration (MIC) values for azole antifungals, while echinocandins and AMB exhibited a narrow range of MIC values, with all strains considered wild-type for the tested polyene and echinocandins. However, when agar diffusion (ellipsometry) was employed for AST, certain strains displayed colonies within the inhibition ellipse, indicating potential resistance. Interestingly, these strains did not respond to AMB treatment and were isolated during AMB treatment (breakthrough). Moreover, the evaluation of AMB minimum fungicidal concentrations (MFCs) indicated that only the strains with colonies inside the ellipse had MFC/MIC ratios ≥ 4, suggesting reduced fungicidal activity. In conclusion, this study confirms the effectiveness of ellipsometry with RPMI-1640 2% glucose agar for detecting AMB resistance in C. lusitaniae. Additionally, the proposed approach of culturing "clear" wells in the microdilution method can aid in uncovering resistant strains. The findings highlight the importance of appropriate AST methods to guide effective treatment strategies for deep-seated candidiasis caused by C. lusitaniae. Further collaborative studies are warranted to validate these findings and improve the detection of AMB clinical resistance.

Resistencia a los azoles en cepas de Aspergilus fumigatus sensu stricto aislados de muestras clínicas respiratorias

Introducción. La aspergilosis invasiva presenta alta mortalidad en pacientes con enfermedades crónicas e inmunocomprometidos. Aspergillus fumigatus sensu stricto (AFSS) es el principal agente etiológico y su tipificación requiere de métodos moleculares. La incidencia incrementada de AI resistentes a los antifúngicos demanda un diagnóstico certero y oportuno. Métodos. Se estudiaron 20 cepas de la micoteca del Instituto de Medicina Tropical - Universidad Nacional Mayor de San Marcos, aisladas de muestras respiratorias e identificadas como Aspergillus fumigatus sensu lato mediante el estudio macroscópico y microscópico. Las cepas fueron referidas a la Universidad Nacional del Litoral para su tipificación mediante una PCR screening para AFSS basada en secuencias del gen CYP51A y el estudio de sensibilidad antifúngica para voriconazol (VOR), itraconazol (ITC), posaconazol (POS), isavuconazol (ISA), anidulafungina (ANF), caspofungina (CSF) y anfotericina B (AMB) obteniendo la concentración inhibitoria mínima (CIM) mediante el protocolo de CLSI M38M51S-Ed3. Resultados. Las 20 cepas fueron identificadas como AFSS. Ninguna de las cepas tuvo una CIM por encima del punto de corte clínico (VOR), ni epidemiológico (ITC, ISA, AMB y CSF). POS fue la droga más potente frente a la colección de cepas evaluadas (media geométrica (GM) de CIM de 0,042 µg/ml). Conclusiones. Todos los aislamientos fueron tipificados como AFSS sensibles a los azoles según los puntos de corte clínico, posaconazol tuvo la mayor actividad antifúngica. Nuestros hallazgos aportan a incrementar la escasa información sobre la etiología y sensibilidad a los antifúngicos de uso clínico de las aspergilosis invasiva en nuestro país.

Introduction. Invasive Aspergillosis (IA) poses a significant threat to patients with chronic diseases and compromised immune systems, with Aspergillus fumigatus sensu stricto (AFSS) being the primary etiological agent. Accurate and timely diagnosis is crucial, particularly given the rising incidence of IA strains resistant to antifungals, necessitating molecular methods for typing. Methods. Twenty strains from Instituto de Medicina Tropical - Universidad Nacional Mayor de San Marcos, mycological collection, previously identified as Aspergillus fumigatus sensu lato through macroscopic and microscopic analysis, were studied. These strains were forwarded to the Universidad Nacional del Litoral for AFSS typing using a PCR screening based on CYP51A gene sequences. Antifungal susceptibility testing was performed for Voriconazole (VOR), Itraconazole (ITC), Posaconazole (POS), Isavuconazole (ISA), Anidulafungin (ANF), Caspofungin (CSF), and Amphotericin B (AMB), obtaining Minimum Inhibitory Concentrations (MICs) according to CLSI M38M51S-Ed3. Results. All 20 strains were identified as AFSS. None of the strains exhibited MICs above the clinical breakpoint (VOR) or the epidemiological cutoffs (ITC, ISA, AMB, and CSF). POS demonstrated the highest potency against the strain collection, with a geometric mean MIC of 0,042 µg/ml. Conclusions. All isolates were classified as azole-sensitive Aspergillus fumigatus sensu stricto (AFSS) based on clinical cutoff points, particularly posaconazole, which exhibited superior antifungal activity. Our findings contribute to augmenting the limited information on the etiology and clinical antifungal sensitivity of IA in our country.

Antifungal susceptibility patterns for Aspergillus, Scedosporium, and Exophiala isolates recovered from cystic fibrosis patients against amphotericin B, and three triazoles and their impact after long-term therapies.

In cystic fibrosis (CF) patients, fungal colonization of the respiratory tract is frequently found. Aspergillus fumigatus, Scedosporium genus, and Exophiala dermatitidis are the most commonly isolated moulds from the respiratory tract secretions of CF patients. The aim of this 5-year surveillance study was to identify trends in species distribution and susceptibility patterns of 212 mould strains identified as Aspergillus spp., Scedosporium spp., and Exophiala spp., isolated from sputum of 63 CF patients who received long-term therapy with itraconazole (ITR) and/or voriconazole (VRC). The Aspergillus isolates were identified as members of the sections Fumigati (n = 130), Flavi (n = 22), Terrei (n = 20), Nigri (n = 8), Nidulantes (n = 1), and Usti (n = 1). Among the 16 species of the genus Scedosporium, 9 were S. apiospermum, 3 S. aurantiacum, and 4 S. boydii. Among the 14 Exophiala species, all were molecularly identified as E. dermatitidis. Overall, 94% (15/16) of Scedosporium spp., 50% (7/14) of E. dermatitidis, and 7.7% (14/182) of Aspergillus spp. strains showed high MIC values (≥8 µg/ml) for at least one antifungal. Particularly, 8.9% (19/212) of isolates showed high MIC values for amphotericin B, 11.7% (25/212) for ITR, 4.2% (9/212) for VRC, and 3.3% (7/212) for posaconazole. In some cases, such as some A. fumigatus and E. dermatitidis isolates recovered from the same patient, susceptibility to antifungal azoles decreased over time. We show that the use of azoles for a long time in CF patients causes the selection/isolation of mould strains with higher MIC values.

The use of azoles for a long time in cystic fibrosis patients causes the selection/isolation of Aspergillus, Scedosporium, and Exophiala species with higher MIC values.

Effect of glyphosate and ciprofloxacin exposure on enteric bacteria of tadpoles / Efecto de la exposición a glifosato y ciprofloxacina en bacterias entéricasde renacuajos

Abstract The high load of agrochemicals and antibiotics present in agricultural aquatic environments represents a risk for wildlife. Since enteric bacteria, which play a key role in the physiological functioning of their hosts, are sensitive to a wide variety of pollutants, their study allows to evaluate the health of organisms. This study aimed to evaluate the effects of commercial formulations of a glyphosate-based herbicide (GBH) and the antibiotic ciprofloxacin (CIP), individually and in mixture, on the bacterial diversity of the intestinal content of common toad (Rhinella arenarum) tadpoles. The diversity of cultivable fast-growing bacteria with low nutritional requirements was evaluated using classic microbiological tests and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification. Bacterial diversity varied among treatments. Taxa diversity increased in the GBH-treated group but decreased in the CIP-treated group. Remarkably, Yersinia spp. and Proteus spp. were only found in the GBH-treated group. The prevalence of Klebsiella spp. and Pseudomonas spp. decreased in the intestinal microbiota of the GBH-CIP-treated group. To our knowledge, this is the first report on the alteration of cultivable enteric bacteria of autochthonous tadpoles due to two pollutants of emerging concern. Our results demonstrate that R. arenarum tadpoles can be used as non-conventional model organisms for environmental pollution monitoring. Our preliminary findings would contribute to understanding how the presence of GBH and CIP in freshwaters may represent a threat to wildlife and human health by causing enteric dysbiosis of part of the bacterial community.

Resumen La alta carga de agroquímicos y antibióticos en los ambientes acuáticos y los agroe-cosistemas representa un riesgo para la vida silvestre. Dado que la microbiota intestinal juega un papel fundamental en el funcionamiento de su hospedador y es sensible a una amplia variedad de contaminantes, su estudio permite evaluar la salud de los organismos. En este trabajo estudiamos los efectos de formulaciones comerciales de un herbicida a base de glifosato (GBH) y del antibiótico ciprofloxacina (CIP), por separado y en mezcla, sobre la diversidad de bacterias intestinales de renacuajos del sapo común (Rhinella arenarum). El estudio de la diversidad de bacterias entéricas cultivables de rápido crecimiento y bajo requerimiento nutricional se llevó a cabo utilizando pruebas microbiológicas clásicas e identificación por espectrometría de masas de tiempo de vuelo por desorción/ionización láser asistida por matriz (MALDI-TOF). La microbiota entérica fue diferente según el tratamiento. El GBH indujo un aumento de la diversidad bacteriana, mientras que la CIP produjo una reducción. Entre estos cambios, destaca la presencia de Yersinia spp. y Proteus spp. solo en el tratamiento con GBH. Además, en el tratamiento GBH-CIP se encontró una disminución en la prevalencia de Klebsiella spp. y Pseudomonas spp. en la microbiota intestinal de los renacuajos. Este es el primer informe sobre la alteración del contenido bacteriano intestinal de renacuajos de R. arenarum producido por dos contaminantes emergentes de preocupación. Demostramos que el renacuajo del sapo común se puede utilizar como un organismo modelo no convencional para el monitoreo de la contaminación ambiental. Estos hallazgos constituyen el primer paso para comprender cómo la presencia de GBH y CIP en aguas dulces puede representar una amenaza para la vida silvestre y la salud humana a través de la disbiosis entérica asociada al efecto sobre la comunidad bacteriana.

Clinical and Demographic Features of Paracoccidioidomycosis in Argentina: A Multicenter Study Analysis of 466 Cases.

Information on paracoccidioidomycosis (PCM) in Argentina is fragmented and has historically been based on estimates, supported only by a series of a few reported cases. Considering the lack of global information, a national multicentric study in order to carry out a more comprehensive analysis was warranted. We present a data analysis including demographic and clinical aspects of a historical series of 466 cases recorded over 10 years (2012-2021). Patients were aged from 1 to 89 years. The general male: female (M:F) ratio was 9.5:1 with significant variation according to the age group. Interestingly, the age range 21-30 shows an M:F ratio of 2:1. Most of the cases (86%) were registered in northeast Argentina (NEA), showing hyperendemic areas in Chaco province with more than 2 cases per 10,000 inhabitants. The chronic clinical form occurred in 85.6% of cases and the acute/subacute form occurred in 14.4% of cases, but most of these juvenile type cases occurred in northwestern Argentina (NWA). In NEA, the incidence of the chronic form was 90.6%; in NWA, the acute/subacute form exceeded 37%. Diagnosis by microscopy showed 96% positivity but antibody detection displays 17% of false negatives. Tuberculosis was the most frequent comorbidity, but a diverse spectrum of bacterial, fungal, viral, parasitic, and other non-infectious comorbidities was recorded. This national multicenter registry was launched in order to better understand the current status of PCM in Argentina and shows the two endemic zones with a highly diverse epidemiology.

Utility evaluation of two molecular methods for Leptospira spp. typing in human serum samples.

Most of the available genotyping methods were applied and evaluated in Leptospira isolates and only few of them in a relevant sample size of blood specimens but not of sera. The objective of this study was to evaluate the utility of one partial 16S rRNA gene sequencing assay (16S rRNA) and an optimized. Multilocus sequence typing scheme (MLST) for Leptospira typing directly in serum samples. Confirmed leptospirosis patients (n = 228) from Argentina (2005-2016) were randomly included. Septicemic-phase serum samples (n = 228) were studied by two genotyping methods. Available immune-phase serum samples of the included patients (n = 159) were studied by MAT to compare serological and molecular results. In culture-proven cases (n = 8), genotyping results between clinical samples and isolates were compared. Typing success rate (TSR) was 21.9% for 16S rRNA and 11.4% for MLST (full allelic profile) and a positive trend in both TSR during the study period was observed. Two species (L. interrogans and L. borgpertesenii) were identified by both methods and MLST assigned 8 different STs. The probable serogroups identified by MLST were coincident with the presumptive infecting serogroups identified by MAT, but with different frequencies. The three serogroups (Canicola, Sejroe and Icterohaemorrhagiae) most frequently identified by MAT were also genotyped by MLST. Typing results via 16S rRNA and MLST in clinical samples and isolates of culture-proven cases, were consistent except for one case. Performance of partial 16S rRNA gene sequencing assay and the optimized MLST scheme directly in sera may increase and improve the knowledge about species and serogroups causing human leptospirosis, especially in countries with low rates of culture sample collection or Leptospira isolation.

Effect of glyphosate and ciprofloxacin exposure on enteric bacteria of tadpoles.

The high load of agrochemicals and antibiotics present in agricultural aquatic environments represents a risk for wildlife. Since enteric bacteria, which play a key role in the physiological functioning of their hosts, are sensitive to a wide variety of pollutants, their study allows to evaluate the health of organisms. This study aimed to evaluate the effects of commercial formulations of a glyphosate-based herbicide (GBH) and the antibiotic ciprofloxacin (CIP), individually and in mixture, on the bacterial diversity of the intestinal content of common toad (Rhinella arenarum) tadpoles. The diversity of cultivable fast-growing bacteria with low nutritional requirements was evaluated using classic microbiological tests and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification. Bacterial diversity varied among treatments. Taxa diversity increased in the GBH-treated group but decreased in the CIP-treated group. Remarkably, Yersinia spp. and Proteus spp. were only found in the GBH-treated group. The prevalence of Klebsiella spp. and Pseudomonas spp. decreased in the intestinal microbiota of the GBH-CIP-treated group. To our knowledge, this is the first report on the alteration of cultivable enteric bacteria of autochthonous tadpoles due to two pollutants of emerging concern. Our results demonstrate that R. arenarum tadpoles can be used as non-conventional model organisms for environmental pollution monitoring. Our preliminary findings would contribute to understanding how the presence of GBH and CIP in freshwaters may represent a threat to wildlife and human health by causing enteric dysbiosis of part of the bacterial community.

Black grain eumycetoma due to Diaporthe ueckerae. Taxonomical update of previous agents of infections due to Diaporthe spp.

A black-grain eumycetoma due to Diaporthe uekerae in a kidney transplant recipient is presented. The isolate was identified by using the newly available NCBI's curated database (rRNA_typestrains/ITS_RefSeq_Fungi) and the NCBI's GenBank + EMBL + DDBI + PDB + RefSeq database. The isolate's antifungal susceptibility was evaluated. The studied isolate showed low MIC values to the eight tested antifungals. Using this updated database, the identities of previous agents of Diaporthe spp. infections were revised.

Candida auris and some Candida parapsilosis strains exhibit similar characteristics on CHROMagarTMCandida Plus.

Candida auris is considered a public health problem due to its resistance and its tendency to cause nosocomial outbreaks. CHROMagarTMCandida Plus has recently been marketed as capable of presumptively identifying C. auris. The objective of this work was to analyze the ability of this new chromogenic medium to differentiate C. auris from other members of the C. haemulonii complex and from other yeasts commonly isolated in clinical practice. A collection of 220 strains including species of the C. haemulonii (n = 83) and C. parapsilosis (n = 80) complexes was studied. The strains were identified by molecular methods and cultured as individual or as mixed aqueous inoculum on CHROMagarTMCandida Plus plates. Colony morphotypes were evaluated at 5 time points. CHROMagarTMCandida Plus was a helpful tool for presumptive identification for C. auris. Better reading results were obtained after 48 hours of incubation at 35°C. It is able to easily differentiate C. auris from other closely related species of the C. haemulonii complex and other yeasts. This chromogenic medium would be also useful as screening and surveillance tool for C. auris colonization. However, we demonstrated that it would be a possible misidentification of C. parapsilosis as C. auris (44.3% showed similar morphotypes). To reduce false positives when it is used in a context of a C. auris outbreak, we propose to supplement the chromogenic medium with 8 µg/ml fluconazole. This modified medium was tested and it clearly differentiate C. parapsilosis from C. auris.

CHROMagarTMCandida Plus is able to differentiate C. auris from other Candida spp., including other species of the C. haemulonii complex. However, 44.3% of the tested C. parapsilosis strains would be misidentified as C. auris. We propose the addition of 8 µg/ml fluconazole to solve this issue.

Usefulness of Sona Aspergillus Galactomannan LFA with digital readout as diagnostic and as screening tool of COVID-19 associated pulmonary aspergillosis in critically ill patients. Data from a multicenter prospective study performed in Argentina.

COVID-19-associated pulmonary aspergillosis (CAPA) incidence varies depending on the country. Serum galactomannan quantification is a promising diagnostic tool since samples are easy to obtain with low biosafety issues. A multicenter prospective study was performed to evaluate the CAPA incidence in Argentina and to assess the performance of the lateral flow assay with digital readout (Sona Aspergillus LFA) as a CAPA diagnostic and screening tool. The correlation between the values obtained with Sona Aspergillus LFA and Platelia® EIA was evaluated. In total, 578 serum samples were obtained from 185 critically ill COVID patients. CAPA screening was done weekly starting from the first week of ICU stay. Probable CAPA incidence in critically ill patients was 10.27% (19/185 patients when LFA was used as mycological criteria) and 9% (9/100 patients when EIA was used as mycological criteria). We found a very good correlation between the two evaluated galactomannan quantification methods (overall agreement of 92.16% with a Kappa statistic value of 0.721). CAPA diagnosis (>0.5 readouts in LFA) were done during the first week of ICU stay in 94.7% of the probable CAPA patients. The overall mortality was 36.21%. CAPA patients' mortality and length of ICU stay were not statistically different from for COVID (non-CAPA) patients (42.11 vs 33.13% and 29 vs 24 days, respectively). These indicators were lower than in other reports. LFA-IMMY with digital readout is a reliable tool for early diagnosis of CAPA using serum samples in critically ill COVID patients. It has a good agreement with Platelia® EIA. LAY SUMMARY: The incidence of COVID-associated pulmonary aspergillosis (CAPA) in critically-ill Argentinian patients was established (10.27%). Serum galactomannan quantification was useful as a screening tool for this mycosis. A good agreement between Platelia® EIA and Sona Aspergillus LFA is reported.

Head-to-head comparison of CLSI, EUCAST, Etest and VITEK®2 results for Candida auris susceptibility testing.

The susceptibility of 31 Candida auris clinical isolates was evaluated by four methods, namely the microdilution reference method according to Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines as well as Etest and VITEK®2. Essential agreement between the two reference methods was 90%. Etest showed a better overall agreement with the reference methods (94% and 81% for CLSI and EUCAST, respectively) than VITEK®2 (70% and 72%, respectively). Discrepancies were found for fluconazole (FLC) and amphotericin B. Considering categorical agreement (CDC tentative breakpoints), the majority of isolates were considered FLC-resistant (93.6% and 80.6% by CLSI and EUCAST, respectively). Furthermore, all isolates were considered susceptible to echinocandins by all methods. Susceptibility results should be interpreted with care if the VITEK®2 system is used to guide therapeutic decisions for C. auris infections.

Functional characterization of monothiol and dithiol glutaredoxins from Leptospira interrogans.

Thiol redox proteins and low molecular mass thiols have essential functions in maintaining cellular redox balance in almost all living organisms. In the pathogenic bacterium Leptospira interrogans, several redox components have been described, namely, typical 2-Cys peroxiredoxin, a functional thioredoxin system, glutathione synthesis pathway, and methionine sulfoxide reductases. However, until now, information about proteins linked to GSH metabolism has not been reported in this pathogen. Glutaredoxins (Grxs) are GSH-dependent oxidoreductases that regulate and maintain the cellular redox state together with thioredoxins. This work deals with recombinant production at a high purity level, biochemical characterization, and detailed kinetic and structural study of the two Grxs (Lin1CGrx and Lin2CGrx) identified in L. interrogans serovar Copenhageni strain Fiocruz L1-130. Both recombinant LinGrxs exhibited the classical in vitro GSH-dependent 2-hydroxyethyl disulfide and dehydroascorbate reductase activity. Strikingly, we found that Lin2CGrx could serve as a substrate of methionine sulfoxide reductases A1 and B from L. interrogans. Distinctively, only recombinant Lin1CGrx contained a [2Fe2S] cluster confirming a homodimeric structure. The functionality of both LinGrxs was assessed by yeast complementation in null grx mutants, and both isoforms were able to rescue the mutant phenotype. Finally, our data suggest that protein glutathionylation as a post-translational modification process is present in L. interrogans. As a whole, our results support the occurrence of two new redox actors linked to GSH metabolism and iron homeostasis in L. interrogans.

[Usefulness of the Sensititre YeastOne® panel to detect Candida species resistant to antifungal drugs]. / Utilidad del panel Sensititre YeastOne® para detectar especies de Candida resistentes a los antifúngicos.

We evaluated the interlaboratory agreement, the essential agreement, and the categorical agreement between the Sensititre YeastOneTM panel and the reference methods M27 4th Edition of the Clinical and Laboratory Standards Institute (CLSI), and the EDef 7.3.1 of the European Committee on Antifungal Susceptibility Testing (EUCAST). We studied 67 Candida strains isolated from different clinical samples and 9 Candida strains with resistance to fluconazole and echinocandins. The highest percentage of interlaboratory agreement was observed with amphotericin B (96.8%), and the lowest percentage with voriconazole (77.2%). Caspofungin showed 5.8% of very major errors when compared with the CLSI reference method. For EUCAST, itraconazole, posaconazole, and anidulafungin showed high percentages of major errors: 17.6%, 18.1%, and 19.6%, respectively. Sensititre YeastOneTM is a reliable alternative, and easy to perform for detecting Candida species resistant to antifungal drugs, with some limitations for echinocandins. Results are comparable to those of the reference methods.

The natural occurring Y129F polymorphism in Rhizopus oryzae (R. arrhizus) Cyp51Ap accounts for its intrinsic voriconazole resistance.

Rhizopus oryzae (heterotypic synonym: R. arrhizus) intrinsic voriconazole and fluconazole resistance has been linked to its CYP51A gene. However, the amino acid residues involved in this phenotype have not yet been established. A comparison between R. oryzae and Aspergillus fumigatus Cyp51Ap sequences showed differences in several amino acid residues. Some of them were already linked with voriconazole resistance in A. fumigatus. The objective of this work was to analyze the role of two natural polymorphisms in the intrinsic voriconazole resistance phenotype of R. oryzae (Y129F and T290A, equivalent to Y121F and T289A seen in triazole-resistant A. fumigatus). We have generated A. fumigatus chimeric strains harboring different R. oryzae CYP51A genes (wild-type and mutants). These mutant R. oryzae CYP51A genes were designed to carry nucleotide changes that produce mutations at Cyp51Ap residues 129 and 290 (emulating the Cyp51Ap protein of azole susceptible A. fumigatus). Antifungal susceptibilities were evaluated for all the obtained mutants. The polymorphism T290A (alone or in combination with Y129F) had no impact on triazole MIC. On the other hand, a > 8-fold decrease in voriconazole MICs was observed in A. fumigatus chimeric strains harboring the RoCYP51Ap-F129Y. This phenotype supports the assumption that the naturally occurring polymorphism Y129F at R. oryzae Cyp51Ap is responsible for its voriconazole resistance phenotype. In addition, these chimeric mutants were posaconazole hypersusceptible. Thus, our experimental data demonstrate that the RoCYP51Ap-F129 residue strongly impacts VRC susceptibility and that it would be related with posaconazole-RoCYP51Ap interaction. LAY SUMMARY: Rhizopus oryzae is intrinsically resistant to voriconazole, a commonly used antifungal agent. In this work, we analyze the role of two natural polymorphisms present in the target of azole drugs. We established that F129 residue is responsible of the intrinsic voriconazole resistance in this species.

Global guideline for the diagnosis and management of rare yeast infections: an initiative of the ECMM in cooperation with ISHAM and ASM.

Uncommon, or rare, yeast infections are on the rise given increasing numbers of patients who are immunocompromised or seriously ill. The major pathogens include those of the genera Geotrichum, Saprochaete, Magnusiomyces, and Trichosporon (ie, basidiomycetes) and Kodamaea, Malassezia, Pseudozyma (ie, now Moesziomyces or Dirkmeia), Rhodotorula, Saccharomyces, and Sporobolomyces (ie, ascomycetes). A considered approach to the complex, multidisciplinary management of infections that are caused by these pathogens is essential to optimising patient outcomes; however, management guidelines are either region-specific or require updating. In alignment with the One World-One Guideline initiative to incorporate regional differences, experts from diverse geographical regions analysed publications describing the epidemiology and management of the previously mentioned rare yeasts. This guideline summarises the consensus recommendations with regards to the diagnostic and therapeutic options for patients with these rare yeast infections, with the intent of providing practical assistance in clinical decision making. Because there is less clinical experience of patients with rare yeast infections and studies on these patients were not randomised, nor were groups compared, most recommendations are not robust in their validation but represent insights by use of expert opinions and in-vitro susceptibility results. In this Review, we report the key features of the epidemiology, diagnosis, antifungal susceptibility, and treatment outcomes of patients with Geotrichum, Saprochaete, Magnusiomyces, and Trichosporon spp infections.

Emergence of Triazole Resistance in Aspergillus spp. in Latin America.

PURPOSE OF REVIEW: Azole resistance in Aspergillus spp. is becoming a public health problem worldwide. However, data about this subject is lacking in Latin American countries. This review focuses in the epidemiology and molecular mechanisms of azole resistance in Aspergillus spp. emphasizing in Latin America. Data on Aspergillus fumigatus stands out because it is the most prevalent Aspergillus spp. pathogen. RECENT FINDINGS: Azole resistance in Aspergillus spp. emergence was linked with intensive use of these antifungals both in the clinical setting and in the environment (as pesticides). Reports on azole-resistant A. fumigatus strains are being constantly published in different countries. Molecular mechanisms of resistance mainly involve substitution in the azole target (CYP51A) and/or overexpression of this gene. However, several other non-CYP51A-related mechanisms were described. Moreover, intrinsically resistant cryptic Aspergillus species are starting to be reported as human pathogens. SUMMARY: After a comprehensive literature review, it is clear that azole resistance in Aspergillus spp. is emerging in Latin America and perhaps it is underestimated. All the main molecular mechanisms of azole resistance were described in patients and/or environmental samples. Moreover, one of the molecular mechanisms was described only in South America. Cryptic intrinsic azole-resistant species are also described.

Infections due to Rare Cryptococcus Species. A Literature Review.

Infections due to rare Cryptococcus species (other than C. neoformans species complex, C. gattii species complex, C. albidus or C. laurentii) are barely reported. The aim of this work is to present a comprehensive literature review of all the papers describing infections due to these species referenced in the main databases (PubMed/MEDLINE, ScienceDirect, Scopus, and Google Scholar). Clinical and epidemiological data together with laboratory findings (identification and antifungal susceptibility) of each isolate were analyzed. Fifty-eight cryptococosis due to rare species were described in 46 papers between 1934-2018. These reports included 16 rare Cryptococcus spp. that were generally associated with nervous system infections and fungemias. Some species are non-capsulated and are not able to grow at 37 °C. Few species were identified by commercially available methods, making internal transcriber spacer (ITS) and D1/D2 regions sequencing mandatory. The most potent antifungal was amphotericin B (although some species showed high MIC values). The studied strains showed high MICs values to 5-fluorocytosine (all >64 µg/mL), echinocandins (all >8 µg/mL), and fluconazole (>80% of the MICs >4 µg/mL). Due to the scarcity of the data and the absence of guidelines for the treatment of these infections, this review could be informative and could help in the diagnosis and treatment of these infections.

Molecular Markers of Antifungal Resistance: Potential Uses in Routine Practice and Future Perspectives.

Antifungal susceptibility testing (AST) has come to establish itself as a mandatory routine in clinical practice. At the same time, the mycological diagnosis seems to have headed in the direction of non-culture-based methodologies. The downside of these developments is that the strains that cause these infections are not able to be studied for their sensitivity to antifungals. Therefore, at present, the mycological diagnosis is correctly based on laboratory evidence, but the antifungal treatment is undergoing a growing tendency to revert back to being empirical, as it was in the last century. One of the explored options to circumvent these problems is to couple non-cultured based diagnostics with molecular-based detection of intrinsically resistant organisms and the identification of molecular mechanisms of resistance (secondary resistance). The aim of this work is to review the available molecular tools for antifungal resistance detection, their limitations, and their advantages. A comprehensive description of commercially available and in-house methods is included. In addition, gaps in the development of these molecular technologies are discussed.

SARS-CoV-2 and Aspergillus section Fumigati coinfection in an immunocompetent patient treated with corticosteroids / Coinfección de SARS-CoV-2 y Aspergillus sección Fumigati en un paciente inmunocompetente tratado con corticosteroides

BACKGROUND: Patients with severe viral pneumonia are likely to receive high-dose immunomodulatory drugs to prevent clinical worsening. Aspergillus species have been described as frequent secondary pneumonia agents in severely ill influenza patients receiving steroids. COVID-19 patients admitted to Intensive Care Unit (ICU) are receiving steroids as part of their treatment and they share clinical characteristics with other patients with severe viral pneumonias. COVID-19 patients receiving steroids should be considered a putative risk group of invasive aspergillosis. CASE REPORT: We are reporting a SARS-CoV-2/Aspergillus section Fumigati coinfection in an elderly intubated patient with a history of pulmonary embolism treated with corticosteroids. The diagnosis was made following the ad hoc definitions described for patients admitted to ICU with severe influenza, including clinical criteria (fever for 3 days refractory to the appropriate antibiotic therapy, dyspnea, pleural friction rub, worsening of respiratory status despite antibiotic therapy and need of ventilator support), a radiological criterion (pulmonary infiltrate) and a mycological criterion (several positive galactomannan tests on serum with ratio ≥0.5). In addition, Aspergillus section Fumigati DNA was found in serum and blood samples. These tests were positive 4 weeks after the patient was admitted to the ICU. The patient received voriconazole and after two month in ICU his respiratory status improved; he was discharged after 6 weeks of antifungal treatment. CONCLUSIONS: Severely ill COVID-19 patients would be considered a new aspergillosis risk group. Galactomannan and Aspergillus DNA detection would be useful methods for Aspergillus infection diagnosis as they allow avoiding the biosafety issues related to these patients

ANTECEDENTES: Los pacientes con neumonía viral grave reciben altas dosis de fármacos inmunomoduladores para prevenir el empeoramiento clínico. Los pacientes con influenza grave que reciben esteroides tienen neumonías secundarias causadas por Aspergillus con una frecuencia relativamente alta. Los pacientes con COVID-19 ingresados en la unidad de cuidados intensivos (UCI) reciben dicha medicación como parte de su tratamiento, y comparten con otro tipo de pacientes muchas de las características clínicas de otras neumonías virales graves. Estos pacientes deberían considerarse como un grupo de riesgo de aspergilosis invasiva. CASO CLÍNICO: Se presenta un caso de coinfección por SARS-CoV-2 y Aspergillus de la sección Fumigati en un paciente intubado de edad avanzada con antecedentes de embolia pulmonar y tratado con corticosteroides. El diagnóstico siguió las definiciones ad hoc descritas para pacientes ingresados en la UCI con gripe grave. El paciente cumplía varios criterios clínicos (fiebre durante 3 días refractaria al tratamiento antibiótico apropiado, disnea, fricción pleural, empeoramiento del estado respiratorio a pesar del tratamiento antibiótico y la necesidad de soporte respiratorio), el criterio radiológico (infiltrado pulmonar) y un criterio micológico (test de galactomanano positivo en suero, (ratio ≥0,5). Además, se detectó ADN de Aspergillus de la sección Fumigati en muestras de suero y sangre del paciente. Estas pruebas fueron positivas 4 semanas después de que el paciente ingresara en la UCI. El paciente recibió tratamiento con voriconazol, y después de 2 meses en la UCI mejoró su estado pulmonar; fue dado de alta después de 6 semanas de tratamiento antifúngico. CONCLUSIONES: Los pacientes gravemente enfermos con COVID-19 deberían considerarse un nuevo grupo de riesgo para la aspergilosis. La detección de galactomanano y ADN de Aspergillus son métodos útiles para el diagnóstico de infección por este hongo al evitar los problemas de bioseguridad en estos pacientes