Genotypic Diversity and Antifungal Susceptibility of Clinical Isolates of Cryptococcus Gattii Species Complex from Argentina.

The aim of this study was to determine the genotypic diversity of 22 Cryptococcus gattii species complex clinical isolates from Argentina and to place these genotypes within the diversity of clinical, veterinary and environmental isolates from Latin America. Mating type and antifungal susceptibility of the isolates were also determined. By URA5-RFLP, nine isolates were identified as molecular type VGI, 10 as VGII, one as VGIII and two as VGIV. Multilocus sequence typing (MSLT), following the International Society for Human and Animal Mycology (ISHAM) consensus MLST scheme, was used to determine the genotypic diversity. Our results suggest that, in Argentina, VGI isolates have low genetic diversity, while VGII isolates have high genetic diversity. Both isolates identified as VGIV by URA5-RFLP were genotyped by MLST as belonging to the currently named VGVI clade. From all isolates, eight sequence types (STs) were unique for Argentina, while five STs have been reported already in other countries, being of high interest the genotypes ST20 and ST7 since they belong to the subtypes VGIIa and VGIIb, respectively, which are associated with hypervirulent strains responsible for outbreaks in North America. To note, geographical analysis showed that some genotypes may be associated with some regions in Argentina. Most isolates were MATα, but we are reporting one isolate MATa for the first time in the country. Antifungal susceptibility tests showed that itraconazole, voriconazole and posaconazole had high activity against all isolates, while amphotericin B, fluconazole and 5-fluorocytosine were the least active drugs against all studied isolates.

Scedosporium aurantiacum: First isolation in Argentina from a previously healthy patient after traumatic inoculation / Scedosporium aurantiacum: primer aislamiento en Argentina de un paciente previamente sano después de una inoculación por un traumatismo

Abstract Scedosporium/Lomentospora species are widely distributed in nature. They are gen-erally saprophytes, but can cause opportunistic infections in immunocompromised patients and occasionally in immunocompetent patients that are difficult to treat due to high lev-els of antifungal resistance. The distribution of Scedosporium/Lomentospora species shows regional differences. Scedosporium boydii and Scedosporium apiospermum are the most fre-quently isolated species in our region, whereas Scedosporium aurantiacum is more common in other regions. We describe the first isolation in Argentina of S. aurantiacum in a vitreous humor infection from a previously healthy patient after traumatic injury in her left eye. Due to the suspicion of fungal endophthalmitis, a mycological study of the vitreous humor was performed. The culture allowed the isolation of S. aurantiacum. The patient was treated with voriconazole with favorable clinic evolution.

Resumen Las especies de Scedosporium/Lamentospora se encuentran ampliamente distribuidas en la naturaleza. En general son saprofitas, pero pueden causar infecciones oportunistas de difícil tratamiento debido a sus altos niveles de resistencia a los antifúngicos en individuos inmunocomprometidos y, ocasionalmente, en personas inmunocompetentes. La distribución de las especies de Scedosporium/Lamentospora muestra diferencias regionales. Scedosporium boydii y S. apiospermum son las especies más frecuentemente aisladas en nuestra región, mientras que en otras S. aurantiacum es más común. Presentamos el primer aislamiento en Argentina de S. aurantiacum de una infección de humor vítreo de un paciente previamente sano que sufrió una lesión traumática. El paciente fue tratado con voriconazol y tuvo una evolución clínica favorable.

Scedosporium aurantiacum: First isolation in Argentina from a previously healthy patient after traumatic inoculation.

Scedosporium/Lomentospora species are widely distributed in nature. They are generally saprophytes, but can cause opportunistic infections in immunocompromised patients and occasionally in immunocompetent patients that are difficult to treat due to high levels of antifungal resistance. The distribution of Scedosporium/Lomentospora species shows regional differences. Scedosporium boydii and Scedosporium apiospermum are the most frequently isolated species in our region, whereas Scedosporium aurantiacum is more common in other regions. We describe the first isolation in Argentina of S. aurantiacum in a vitreous humor infection from a previously healthy patient after traumatic injury in her left eye. Due to the suspicion of fungal endophthalmitis, a mycological study of the vitreous humor was performed. The culture allowed the isolation of S. aurantiacum. The patient was treated with voriconazole with favorable clinic evolution.

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.

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

Resumen Evaluamos las concordancias interlaboratorio, esencial y categórica entre el panel Sensititre YeastOne™ y los métodos de referencia correspondientes al M27 4.a ed. (Clinical and Laboratory Standards Institute [CLSI]) y al EDef 7.3.1 (European Committee on Antifungal Susceptibility Testing [EUCAST]). Estudiamos 67 cepas de Candida de distintas muestras clí nicas y un panel de 9 cepas resistentes a fluconazol y equinocandinas. El mayor porcentaje de concordancia interlaboratorio se observó con anfotericina B (96,8%) y el menor porcen taje con voriconazol (77,2%). La caspofungina mostró un 5,8% de discrepancias muy mayores con el método de referencia del CLSI. Con el del EUCAST, el itraconazol, el posaconazol y la anidulafungina mostraron porcentajes de discrepancias mayores: el 17,6, el 18,1 y el 19,6%, respectivamente. El panel Sensititre YeastOne™ es una alternativa confiable y fácil de usar, que permite detectar especies de Candida resistentes a los antifúngicos, con algunas limitaciones para las equinocandinas. Los resultados son equiparables a los de los métodos de referencia.

Abstract We evaluated the interlaboratory agreement, the essential agreement, and the cate- gorical agreement between the Sensititre YeastOne™ panel and the reference methods M27 4th Edition of the Clinical and Laboratory Standards Institute (CLSI), and the EDef 7.3.1 of the Euro- pean 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 amp- hotericin 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 YeastOne™ 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.

Equine rhinosporidiosis in Buenos Aires, Argentina

Abstract Rhinosporidiosis is caused by Rhinosporidium seeberi, a parasitic organism of the family Rhinosporideacea family, class Micomycetozoa. The disease is endemic in India; however, some cases were reported in Europe, Africa, North America, and South America. The aim of the present study is to report three cases of rhinosporidiosis in wild horses in different cities of Buenos Aires province, Argentina. We confirm the presence of R. seeberi in the analyzed samples using histopathological and PCR sequencing techniques.

Resumen La rinosporidiosis es una enfermedad causada por Rhinosporidium seeberi, un organismo parasitario clasificado en la familia Rhinosporideacea, clase Micomycetozoa. Es una enfermedad endémica de la India, pero se notificaron algunos casos en Europa, África, América del Norte y América del Sur. El objetivo del presente estudio fue describir tres casos de rinosporidiosis en caballos de vida libre en diferentes ciudades de la provincia de Buenos Ares, Argentina. Confirmamos la presencia de R. seeberi en las muestras analizadas utilizando técnicas histopatológicas, PCR y secuenciación.

[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.

Equine rhinosporidiosis in Buenos Aires, Argentina.

Rhinosporidiosis is caused by Rhinosporidium seeberi, a parasitic organism of the family Rhinosporideacea family, class Micomycetozoa. The disease is endemic in India; however, some cases were reported in Europe, Africa, North America, and South America. The aim of the present study is to report three cases of rhinosporidiosis in wild horses in different cities of Buenos Aires province, Argentina. We confirm the presence of R. seeberi in the analyzed samples using histopathological and PCR sequencing techniques.

Effectiveness of the National Quality Control Program in Mycology of Argentina and performance evaluation of participating laboratorios / Efectividad del Programa Nacional de Control de Calidad en Micología de Argentina y evaluación del desempeño de los laboratorios participantes

Abstract The National Quality Control Program in Mycology (PNCCM) of Argentina was establishedin 1996 to improve the quality of the mycological diagnosis, to help establish and to setup standardized procedures and continuous training of laboratory staff. The aim of this studywas to assess the effectiveness of the PNCCM in the 1996---2018 period. Data from the NationalMycology Laboratory Network (NMLN) and PNCCM database was used to estimate the increasein the number of controlled laboratories and jurisdictions, the percentage of participation, theimprovement in the quality of results and the adherence to the program. Satisfaction surveyswere performed to assess user satisfaction. The number of controlled laboratories increasedfrom 29 to 146; participation increased from 49% to 93% and general adherence was 72% inthe evaluated period (1996---2018). Improvement in the quality of the results was 15% for lowcomplexity samples; 7% for intermediate complexity samples and 14% for the identification ofhigh complexity strains. Up to 84% of the users consider the PNCCM to be ''very good'' and 16%''satisfactory''. These results show the importance of the PNCCM, which is widely accepted bymycological diagnostic laboratories from Argentina.

Resumen En 1996 se creó el Programa Nacional de Control de Calidad en Micología (PNCCM)de Argentina con el objetivo de mejorar la calidad del diagnóstico micológico, colaborar enel establecimiento de procedimientos estandarizados en aquellos laboratorios que carecen deellos y contribuir a la capacitación continua del personal.El objetivo de este estudio fue evaluar la efectividad del PNCCM en el período 1996-2018.Se utilizaron los datos de la base de la Red Nacional de Laboratorios de Micología (RNLM) ydel PNCCM para estimar el aumento en el número de laboratorios controlados y el porcentajede participación, la mejora de la calidad de los resultados y la adhesión al programa. Paraevaluar el grado de satisfacción de los usuarios, se analizaron las encuestas de satisfacción delos participantes. En el período evaluado, el número de laboratorios controlados aumentó de 29a 146, la participación aumentó de 49% a 93% y la adherencia general de los participantes fue del72%. La mejora de la calidad de los resultados de los laboratorios fue del 15% para muestras debaja complejidad, 7% para muestras de complejidad intermedia y 14% para la identificación decepas de alta complejidad. El 84% de los usuarios considera que el PNCCM es muy bueno y el 16%que es satisfactorio. Estos resultados evidencian la importancia del PNCCM, que es ampliamenteaceptado por los laboratorios que realizan diagnóstico micológico en nuestro país.

Scedosporium spp. from Clinical Setting in Argentina, with the Proposal of the New Pathogenic Species Scedosporium americanum.

Species of the genus Scedosporium (family Microascaceae, phylum Ascomycota) are responsible for a wide range of opportunistic human infections, and have a low susceptibility to most antifungal drugs. It is well known that the pattern of Scedosporium species distribution varies according to geographic region. To assess the diversity of Scedosporium species in Argentina involved in human infections, we carried out a retrospective study reviewing 49 strains from clinical samples sent for diagnosis to the National Clinical Mycology Reference Laboratory between 1985 and 2019. Then, a phenotypic characterization, a phylogenetic study and and in vitro susceptibility test to antifungals were carried out. An analysis of combined nucleotide sequences dataset of the internal transcribed spacer of the ribosomal DNA (ITS) and of a fragment of the ß-tubulin gene (BT2) demonstrated that 92 % of the strains belonged to the species S. boydii, S. apiospermum and S. angustum, all them pertaining to S. apiospermum species complex. However, two strains (4%) were identified as S. aurantiacum, a species never reported in clinical settings in the Americas'. Surprisingly, one of them displayed a polycytella-like conidiogenesis, up to date only reported for S. apiospermum. In addition, the strain DMic 165285 was phylogenetically located far away from the rest of the species, so is proposed as the novel species Scedosporium americanum. On the other hand, from all seven antifungals tested, voriconazole and posaconazole were the most active drugs against Scedosporium spp.

Frequency of sensitization to the individual fragrances of fragrance mix I and II according to the factors included in the MOAHLFA index.

BACKGROUND: Fragrances constitute the second most frequent cause of allergic contact dermatitis in Spain. OBJECTIVES: To determine the rate of sensitization to the individual fragrances of fragrance mix (FM) I and FM II for each of the demographic and clinical factors included in the MOAHLFA (male, occupational dermatitis, atopic dermatitis, hand dermatitis, leg dermatitis, facial dermatitis, age) index. METHODS: We conducted a 5-year retrospective study in 23 Spanish centres. We identified the patients who had undergone patch testing with a specific fragrance series after reacting positively to fragrance markers in a baseline series. We obtained the MOAHLFA index items in this population, then calculated for each demographic and clinical factor the frequencies of sensitization to the individual fragrances of FM I and FM II. RESULTS: A specific fragrance series was patch tested in 1013 patients. The most frequent allergens in men, women, children, and retired people were Evernia prunastri (16%), geraniol (16.6%), isoeugenol (17.9%), and geraniol (22.4%), respectively. Citral (20.5%) and hydroxyisohexyl 3-cyclohexene carboxaldehyde (HICC) (14.5%) were the most common allergens in occupational eczemas and were also associated with a large proportion of hand and facial dermatitis. CONCLUSIONS: Frequency of sensitization to the individual fragrances of FM I and FM II varies with age, sex, affected body region, and history of occupational or atopic dermatitis.

Effectiveness of the National Quality Control Program in Mycology of Argentina and performance evaluation of participating laboratories.

The National Quality Control Program in Mycology (PNCCM) of Argentina was established in 1996 to improve the quality of the mycological diagnosis, to help establish and to set up standardized procedures and continuous training of laboratory staff. The aim of this study was to assess the effectiveness of the PNCCM in the 1996-2018 period. Data from the National Mycology Laboratory Network (NMLN) and PNCCM database was used to estimate the increase in the number of controlled laboratories and jurisdictions, the percentage of participation, the improvement in the quality of results and the adherence to the program. Satisfaction surveys were performed to assess user satisfaction. The number of controlled laboratories increased from 29 to 146; participation increased from 49% to 93% and general adherence was 72% in the evaluated period (1996-2018). Improvement in the quality of the results was 15% for low complexity samples; 7% for intermediate complexity samples and 14% for the identification of high complexity strains. Up to 84% of the users consider the PNCCM to be "very good" and 16% "satisfactory". These results show the importance of the PNCCM, which is widely accepted by mycological diagnostic laboratories from Argentina.

Feohifomicosis por Exserohilum rostratum en un paciente pediátrico con leucemia linfoblástica aguda postransplante de médula ósea / Phaeohyphomycosis by Exserohilum rostratum in a pediatric patient with acute lymphoblastic leukemia after bone marrow transplantation

Resumen El género Exserohilum corresponde a hongos dematiáceos, la mayoría fitopatógenos, saprobios, de los cuales solo tres especies serían patógenas para el hombre: Exserohilum rostratum, Exserohilum longirostratum y Exserohilum mcginnisii. Se han reportado infecciones localizadas y sistémicas causadas por estos agentes, tanto en pacientes inmunocompetentes como inmunosuprimidos. Se presenta un caso de infección cutánea por E. rostratum en un paciente pediátrico con inmunocompromiso.

Abstract The genus Exserohilum consists of dematiaceous or darkly pigmented fungi. Most of the species included in this genus are phytopathogens, saprobes and only three of these species would be pathogenic to humans: Exserohilum rostratum, Exserohilum longirostratum and Exserohilum mcginnisii. Localized and systemic infections have been reported both in immunocompetent and immunosuppressed patients. A clinical case of cutaneous infection by E. rostratum in an immunocompromised pediatric patient is presented in this study.

[Phaeohyphomycosis by Exserohilum rostratum in a pediatric patient with acute lymphoblastic leukemia after bone marrow transplantation]. / Feohifomicosis por Exserohilum rostratum en un paciente pediátrico con leucemia linfoblástica aguda postransplante de médula ósea.

The genus Exserohilum consists of dematiaceous or darkly pigmented fungi. Most of the species included in this genus are phytopathogens, saprobes and only three of these species would be pathogenic to humans: Exserohilumrostratum, Exserohilumlongirostratum and Exserohilummcginnisii. Localized and systemic infections have been reported both in immunocompetent and immunosuppressed patients. A clinical case of cutaneous infection by E. rostratum in an immunocompromised pediatric patient is presented in this study.'

First case report of cryptococcosis due to Cryptococcus decagattii in a pediatric patient in Argentina.

We report the first case of cryptococcosis due to Cryptococcus decagattii in an immunocompetent pediatric patient from an indigenous community in Argentina with a successful outcome. Two isolates (blood, cerebrospinal fluid) were genotyped by restriction fragment length polymorphism of the orotidine monophosphate pyrophosphorylase (URA5) gene as VGIV and identified by multi-locus sequence typing as C. decagattii. Matrix-assisted laser desorption/ionization time of flight mass spectrometry identification indicated genotype VGIII. The minimum inhibitory concentration of amphotericin B, fluconazole, itraconazole, and voriconazole was determined (cerebrospinal fluid: 0.25, 16, 0.12, and 0.12, blood: 0.25, 4, 0.12, and 0.06, respectively, all in mg/L).

First bloodstream infection due to Prototheca zopfii var. hydrocarbonea in an immunocompromised patient.

Here we describe a bloodstream infection due to P. zopfii var. hydrocarbonea in a patient with acute lymphoblastic leukemia. Identification was performed by DNA sequencing of the D1/D2 domain of 26s ribosomal DNA and by MALDI-TOF MS technique. Antifungal susceptibility tests against amphotericin B, fluconazole, itraconazole, and voriconazole showed the following MIC values, respectively: 0.25 mg/L, 128 mg/L, 0.064 mg/L, and 0.125 mg/L. The patient received amphotericin B treatment with a successful outcome.

Reidentification and antifungal susceptibility profile of Candida guilliermondii and Candida famata clinical isolates from a culture collection in Argentina.

The aim of this work was to reidentify strains previously identified as Candida guilliermondii and Candida famata by conventional phenotypic methods conserved in a culture collection from Argentina using ribosomal DNA sequencing, ACT1 gene sequencing, and matrix-assisted laser desorption ionization - time of flight mass spectrometry (MALDI-TOF MS). In addition, we performed antifungal susceptibility tests of eight antifungal drugs commonly used in clinical treatment. We identified 68 isolates belonging to the Candida guilliermondii species complex (59 C. guilliermondii, 8 C. fermentati, and 1 Candida carpophila), 16 isolates belonging to the Candida famata species complex (8 C. famata, 6 Debaryomyces nepalensis, 1 Debaryomyces fabryi, and 1 Debaryomyces tyrocola). Although sequencing of ITS region was able to identify C. guilliermondii and D. nepalensis isolates, sequencing of ACT1 gene seems to be the most appropriate technique for differentiation between C. fermentati and C. carpophila and between members of the C. famata species complex others than D. nepalensis. MALDI-TOF MS has a good potential for the identification of these yeasts, particularly in clinical laboratories since is a rapid and easy to perform technique. Here, we report the first isolation of D. tyrocola from a human patient and the first isolation of D. nepalensis from lungs and blood of human patients. Finally, correct identification and determination of antifungal susceptibility of those closely related species could be a useful tool for clinicians to choose the most effective antifungal treatment.