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.

First report of phaeohyphomycosis caused by Phialophora americana in a domestic cat from Argentina.

Case summary: A 10-year-old male neutered domestic shorthair cat from Quilmes (Province of Buenos Aires, Argentina) presented at the Infectious Diseases and Parasitology Unit with a hyperpigmented nodule of 5 cm diameter on the nasal plane with a small ulceration of more than 1 year's evolution. A scaly and hyperpigmented alopecic lesion of 3 cm in diameter was found on the lower edge of the tail. The patient was under immunosuppressive therapy with corticosteroids for lymphoplasmacytic duodenitis. Samples of the lesion present on the nasal plane were taken under a surgical procedure. In the wet mount preparations, pigmented irregular hyphae were observed. They developed dark colonies when cultured on Sabouraud medium. On micromorphology, structures compatible with Phialophora species were identified. PCR and sequencing of ITS (ITS1-5.8S-ITS2) confirmed Phialophora americana as the etiologic agent. A therapeutic scheme that included a combination of itraconazole oral solution (1.5 mg/kg PO q12h) with terbinafine (30 mg/kg PO q24h) was indicated for a period of 10 months. The patient died of complications resulting from its underlying disease. Relevance and novel information: As far as the authors are aware, this is the first study to report P americana as an etiologic agent of phaeohyphomycosis in cats. In this case study, the species was identified using molecular tests.

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.

Dermatophytosis caused by Trichophyton benhamiae in a dog.

Trichophyton benhamiae was diagnosed in a 9-year-old female dog by histopathological evaluation, fungal culture and confirmation by sequencing of the internal transcribed spacer region of ribosomal DNA. Successful therapy was achieved with itraconazole, bathing with miconazole and chlorhexidine shampoo, and topical application of sodium hypochlorite as a rinse.

Trichophyton benhamiae a été diagnostiqué chez une chienne de 9 ans par examen histopathologique, culture fongique et confirmation par séquençage de la région ITS (internal trasbcriber spacer) de l'ADN ribosomique. Une guérison thérapeutique a été obtenue par de l'itraconazole, des shampooings de miconazole et chlorhexidine et application topique d'hypochlorite de sodium en rinçage.

Trichophyton benhamiae foi diagnosticado em uma cadela de 9 anos através de avaliação histopatológica, cultura de fungos e confirmação por sequenciamento da região espaçadora transcrita interna do DNA ribossomal. Sucesso terapêutico foi obtido com o uso de itraconazol, banho com shampoo à base de miconazol e clorexidina e aplicação tópica de hipoclorito de sódio.

Se diagnosticó infección por Trichophyton benhamiae en una perra de 9 años mediante evaluación histopatológica, cultivo de hongos y confirmación mediante secuenciación de la región espaciadora transcrita interna del DNA ribosómico. Se logró un tratamiento exitoso con itraconazol, baños con champú de miconazol y clorhexidina y aplicación tópica de hipoclorito de sodio como enjuague.

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.

Rhizopus microsporus Infections Associated with Surgical Procedures, Argentina, 2006-2014.

Rhizopus spp. fungi are ubiquitous in the environment and a rare but substantial cause of infection in immunosuppressed persons and surgery patients. During 2005-2017, an abnormally high number of Rhizopus infections in surgery patients, with no apparent epidemiologic links, were reported in Argentina. To determine the likelihood of a common source of the cluster, we performed whole-genome sequencing on samples collected during 2006-2014. Most isolates were separated by >60 single-nucleotide polymorphisms, and we found no evidence for recombination or nonneutral mutation accumulation; these findings do not support common source or patient-to-patient transmission. Assembled genomes of most isolates were ≈25 Mbp, and multiple isolates had substantially larger assembled genomes (43-51 Mbp), indicative of infections with strain types that underwent genome expansion. Whole-genome sequencing has become an essential tool for studying epidemiology of fungal infections. Less discriminatory techniques may miss true relationships, possibly resulting in inappropriate attribution of point source.

Mycosis Due to Tropicoporus tropicalis (= Inonotus tropicalis) in a Domestic Dog.

Mycelial basidiomycetes rarely produce mycoses in animals including humans. We report a case of a 9-year-old female mongrel dog with lesions in the prescapular lymph nodes. The histopathology of a lymph node sample showed flexuous septate hyphae, and a sterile mold grew in culture from that specimen. DNA sequencing of the ITS region allowed us to identify the fungus as Tropicoporus tropicalis. The dog was treated with itraconazole, but it was euthanized six months later due to an unfavorable clinical outcome. Tropicoporus tropicalis is an infrequent pathogen of pets, and the use of molecular tools is needed for its identification. Animal infections due to T. tropicalis were not previously been reported in Argentina.

Osteomielitis por mucorales (OMM) asociada a artroscopia de reparación de ligamento cruzado anterior: investigación epidemiológica y recomendaciones para su prevención / Mucor osteomyelitis after arthroscopic repair of ACL. Epidemiological investigation

OBJETIVO: Describir la investigación epidemiológica de osteomielitis por Mucorales (OMM) post reparación artroscópica de LCA (RA-LCA) en Argentina. MATERIAL Y MÉTODO: 1) Revisión de los casos; 2) Relevamiento de 3 instituciones; 3) Cultivo micológico de materiales quirúrgicos; 4) Encuesta a instrumentadoras; 5) Secuenciación de las cepas de Rhizopus y 6) Redacción de recomendaciones. RESULTADOS: Del 2005 al 2017 se identificaron 40 casos de OMM (Rhizopus sp.) post reparación artroscópica de LCA en pacientes inmunocompetentes de 12 jurisdicciones de Argentina. El diagnóstico fue por cultivo (22/31), y por anatomía patológica (9). La edad promedio fue 29 años. El 84% de 38 casos eran varones. Intervinieron 13 ortopedias. El implante fue importado en 8/20 casos y nacional en 12. En las 3 instituciones se observó: manejo inadecuado del aire de quirófano, variabilidad en la limpieza del artroscopio, en el taladro utilizado, y en el manejo de materiales que llegan de las ortopedias y falta de trazabilidad de los implantes. Los cultivos micológicos de los materiales fueron negativos. La encuesta a instrumentadores confirmó los hallazgos de los relevamientos. La secuenciación de las cepas de Rhizopus demostró predominio de policlonalidad. CONCLUSIÓN: La OMM es una complicación posible luego de la RA-LCA en instituciones privadas de Argentina. No se identificó un origen único. Se detectaron múltiples prácticas que favorecen la contaminación de la cirugía con hongos filamentosos (manejo del aire de quirófano, del artroscopio, de los materiales provenientes de ortopedia, etc.). En base a estos hallazgos la Asociación Argentina de Artroscopía sugiere medidas de prevención. Implicancia clínica: Prevención de osteomielitis por Mucorales post- cirugía artroscópica para ligamento cruzado anterior. Tipo de estudio: Serie de casos. Nivel de Evidencia: IV.

OBJECTIVE: To describe the epidemiological investigation of Mucor osteomyelitis (MO) after arthroscopic repair of ACL (ARACL) in Argentina. MATERIAL Y METHODS: 1) Review of cases; 2) Survey of 3 institutions; 3) Mycological culture of surgical materials; 4) Survey of instrumentists; 5) Sequencing of Rhizopus strains and 6) Writing of recommendations. RESULTS: From 2005 to 2017, 40 cases of MO (Rhizopus sp.) Post AR-ACL were identified in immunocompetent patients from 12 jurisdictions of Argentina. The diagnosis was made by culture (22/31), and by pathology (9). The average age was 29 years. 84% of 38 cases were male. Thirteen orthopedics intervened. The implant was imported in 8/20 cases and national in 12. In the 3 institutions it was observed: inadequate handling of the operating room air, variability in the cleaning of the arthroscope, in the drill used, and in the handling of materials that come from the orthopedics and lack of traceability of the implants. The mycological cultures of the materials were negative. The survey of instrumentists confirmed the findings of the surveys. The sequencing of Rhizopus strains showed a predominance of polyclonality. CONCLUSION: MO is a possible complication after AR-ACL in private institutions in Argentina. A unique origin was not identified. Multiple practices that favor the contamination of surgery with filamentous fungi (handling of operating room air, arthroscope, materials from orthopedics, etc.) were detected. Based on these findings, the Argentine Association of Arthroscopy suggests prevention measures. Clinical relevance: Prevention of Mucor osteomyelitis after arthroscopic surgery for anterior cruciate ligament. Type study: Cases series. Level of evidence: IV.

kMucormicosys due to Saksenaea vasiformis in a dog.

A 2-year-old female Border collie was examined for dermatitis with a partial alopecic zone around her left front member. Six months later the lesion became swollen, alopecic with ulcerated areas. Microscopy analysis of samples showed numerous non-septate, branching, thin-walled and irregular shaped hyphal elements. Fungal cultures and molecular studies identified Saksenaea vasiformis. Treatments with griseofulvin, itraconazole and surgical debridement were used, however, fourteen months later the dog was euthanatized because of the unfavorable clinical outcome.

Keratitis caused by moulds in Santa Lucía Ophthalmology Hospital in Buenos Aires, Argentina / Queratitis causada por hongos miceliares en el Hospital Oftalmológico Santa Lucía de Buenos Aires, Argentina(AU)

Background. Mycotic keratitis by moulds (MKM) is an important cause for corneal blindness and usually carries an unfavorable prognosis. Aims. This study describes the risk factors and demographic and microbiological features of all MKM cases in Santa Lucía Ophthalmology Hospital during a period of 6 years. Methods. A prospective study was performed for all MKM cases diagnosed between October 2007 and September 2013. Results. Among 157 diagnosed cases, direct microscopic examination and culture were positive in 97 and 96% of the cases respectively. MKM represents 17% of all microbiologically confirmed corneal abscesses. No significant differences were detected in annual MKM frequencies across the study period, suggesting that MKM incidence remains constant over time. A male-to-female ratio was observed (2.8:1); the most affected age groups ranged from 31 to 40 years old (males) and 61-70 years old (females). The most frequent predisposing factor was trauma (40%) followed by the use of contact lenses (9%), herpetic abscesses (5%) and diabetes (4%). The predominant genera were Fusarium (66%), Aspergillus (10%), Curvularia (6%) and Alternaria (4%). The most frequent agent was Fusarium solani species complex (52%). More than two-thirds of the cases were produced by only 3 species or complexes. However, at least 29 different species were detected in the remaining cases. This is the first report of Pholiota sp. as causative agent of human MKM. Conclusions. Argentina lacks extensive epidemiological and clinical data on MKM. This six-year study performed in Argentina is a first step leading to a better understanding of MKM epidemiology in our country (AU)

Antecedentes. La queratitis micótica causada por hongos miceliares (MKM) es una causa importante de ceguera corneal y presenta un pronóstico desfavorable. Objetivos. Describir los factores de riesgo y las características demográficas y microbiológicas de las MKM diagnosticadas en el Hospital Oftalmológico Santa Lucía durante 6 años. Métodos. Se llevó a cabo un estudio prospectivo de todos los casos de MKM diagnosticados entre octubre de 2007 y septiembre de 2013. Resultados. Se diagnosticaron 157 casos de MKM: un 97% con examen directo positivo y un 96% con cultivo positivo. Las MKM representaron el 17% de todos los abscesos corneales microbiológicamente confirmados. No se encontraron diferencias significativas en esta proporción durante los seis años de estudio, lo que demuestra que la incidencia permaneció constante. La proporción varones:mujeres fue 2,8:1; las edades más afectadas fueron 31-40 años en los varones y 61-70 años en las mujeres. El factor predisponente más común fue el traumatismo (40%), seguido del uso de lentes de contacto (9%), abscesos herpéticos (5%) y diabetes (4%). Los géneros predominantes fueron Fusarium (66%), Aspergillus (10%), Curvularia (6%) y Alternaria (4%). Los agentes más frecuentes pertenecían al complejo de especies Fusarium solani (52%). Más de dos tercios de los casos fueron causados únicamente por 3 especies, o complejos de especies, pero en los restantes se detectaron al menos otras 28 especies. Este es el primer informe referente a Pholiota sp. como agente causal de MKM humana. Conclusiones. Argentina carece de datos epidemiológicos y clínicos consolidados sobre MKM. Este estudio contribuye a una mejor comprensión de la epidemiología de esta enfermedad en este país (AU)

Keratitis caused by moulds in Santa Lucía Ophthalmology Hospital in Buenos Aires, Argentina.

BACKGROUND: Mycotic keratitis by moulds (MKM) is an important cause for corneal blindness and usually carries an unfavorable prognosis. AIMS: This study describes the risk factors and demographic and microbiological features of all MKM cases in Santa Lucía Ophthalmology Hospital during a period of 6 years. METHODS: A prospective study was performed for all MKM cases diagnosed between October 2007 and September 2013. RESULTS: Among 157 diagnosed cases, direct microscopic examination and culture were positive in 97 and 96% of the cases respectively. MKM represents 17% of all microbiologically confirmed corneal abscesses. No significant differences were detected in annual MKM frequencies across the study period, suggesting that MKM incidence remains constant over time. A male-to-female ratio was observed (2.8:1); the most affected age groups ranged from 31 to 40 years old (males) and 61-70 years old (females). The most frequent predisposing factor was trauma (40%) followed by the use of contact lenses (9%), herpetic abscesses (5%) and diabetes (4%). The predominant genera were Fusarium (66%), Aspergillus (10%), Curvularia (6%) and Alternaria (4%). The most frequent agent was Fusarium solani species complex (52%). More than two-thirds of the cases were produced by only 3 species or complexes. However, at least 29 different species were detected in the remaining cases. This is the first report of Pholiota sp. as causative agent of human MKM. CONCLUSIONS: Argentina lacks extensive epidemiological and clinical data on MKM. This six-year study performed in Argentina is a first step leading to a better understanding of MKM epidemiology in our country.

Genotipificación de aislamientos clínicos de Aspergillus flavus y su relación con aislamientos ambientales de un centro oncohematológico / Genotyping of clinical isolates of Aspergillus flavus and its relationship with environmental isolates of an oncohematological center

Antecedentes. Durante un período de 4 meses, y mientras se llevaba a cabo un muestreo ambiental de aire, se diagnosticaron 2 casos de aspergilosis por Aspergillus flavus en un centro oncohematológico de Buenos Aires, Argentina. Objetivos. Conocer la variabilidad y la relación genética entre los aislamientos clínicos y los ambientales obtenidos en el centro oncohematológico. Métodos. Se utilizaron 2 técnicas de genotipificación con diferente poder discriminatorio (RAPD y AFLP). Una matriz de similitud genética fue calculada usando el método de Jaccard y fue la base para la construcción de un dendrograma por el método de UPGMA. Se estimó el nivel de variabilidad genética por medio del porcentaje de loci polimórficos, número de alelos efectivos y heterocigosidad esperada, y el índice de asociación (IA). Resultados. El dendrograma mostró que los aislamientos de A. flavus recuperados de los pacientes no se relacionaron genéticamente con los del ambiente nosocomial. Los valores más altos de diversidad genética correspondieron a los aislamientos ambientales. El IA estimado para todos los aislamientos sugiere eventos de recombinación. Conclusiones. Los pacientes 1 y 2 no fueron infectados con los aislamientos obtenidos del ambiente hospitalario. Los aislamientos clínicos y ambientales de A. flavus mostraron alta variabilidad genética entre ellos(AU)

Background. During 4 months, and while conducting an environmental sampling of air, 2 cases of aspergillosis by Aspergillus flavus (A. flavus) were diagnosed at an oncohematological center in Buenos Aires, Argentina. Aims. The aim of this study was to know the variability and the genetic relationship between the clinical and environmental isolates, obtained in the oncohematological center. Methods. Two genotyping techniques of different discriminatory power (RAPD and AFLP) were used. A genetic similarity matrix was calculated using Jaccard method and was the basis for the construction of a dendrogram by UPGMA. The level of genetic variability was assessed by measuring the percentage of polymorphic loci, number of effective allele, expected heterocygozity and association index test (IA). Results. The dendrogram reveals that the A. flavus isolates recovered from the patients were not genetically related to those gotten from the rooms occupied by the patients. The environmental isolates had higher values of genetic diversity than the clinical isolates. The IA estimated for all the isolates suggest that recombination events occurred. Conclusions. Patients 1 and 2 were not infected with isolates from the nosocomial environment. Clinical and environmental isolates of A. flavus showed high genetic variability among them(AU)

[Genotyping of clinical isolates of Aspergillus flavus and its relationship with environmental isolates of an oncohematological center]. / Genotipificación de aislamientos clínicos de Aspergillus flavus y su relación con aislamientos ambientales de un centro oncohematológico.

BACKGROUND: During 4 months, and while conducting an environmental sampling of air, 2 cases of aspergillosis by Aspergillus flavus (A. flavus) were diagnosed at an oncohematological center in Buenos Aires, Argentina. AIMS: The aim of this study was to know the variability and the genetic relationship between the clinical and environmental isolates, obtained in the oncohematological center. METHODS: Two genotyping techniques of different discriminatory power (RAPD and AFLP) were used. A genetic similarity matrix was calculated using Jaccard method and was the basis for the construction of a dendrogram by UPGMA. The level of genetic variability was assessed by measuring the percentage of polymorphic loci, number of effective allele, expected heterocygozity and association index test (I(A)). RESULTS: The dendrogram reveals that the A. flavus isolates recovered from the patients were not genetically related to those gotten from the rooms occupied by the patients. The environmental isolates had higher values of genetic diversity than the clinical isolates. The I(A) estimated for all the isolates suggest that recombination events occurred. CONCLUSIONS: Patients 1 and 2 were not infected with isolates from the nosocomial environment. Clinical and environmental isolates of A. flavus showed high genetic variability among them.

In vitro interactions of antifungal agents against clinical isolates of Fusarium spp.

The in vitro activities of amphotericin B (AMB), itraconazole (ITC), voriconazole (VCZ) and terbinafine (TBF) alone and in the combinations AMB+VCZ, TBF+ITC and TBF+VCZ were evaluated against 29 clinical isolates of Fusarium spp. (15 Fusarium solani, 7 Fusarium oxysporum, 2 Fusarium decemcellulare, 2 Fusarium dimerum and 3 other Fusarium spp.). Minimum inhibitory concentrations were determined using the method of the Clinical and Laboratory Standards Institute and the interaction activity was calculated using the fractional inhibitory concentration index. The four antifungal drugs tested alone showed very limited activity against most of the isolates. In contrast, the combination TBF+VCZ showed synergy for 21 isolates. The combination AMB+VCZ showed synergism for only five strains. No interaction or antagonism was observed among the remaining strains. TBF+ITC showed no interaction for 18 strains. The in vitro antifungal activity of the drugs alone and in combination varied for different species. These results corroborate previous in vitro studies in which the combination TBF+VCZ showed synergy against Fusarium spp., although further studies are needed to elucidate its potential usefulness for therapy.