Penicillium digitatum, First Clinical Report in Chile: Fungal Co-Infection in COVID-19 Patient.

Penicillium digitatum is one of the most important phytopathogens. It causes deterioration and rotting of citrus fruits, generating significant economic losses worldwide. As a human pathogen, it is extremely rare. We present a case of pulmonary co-infection in a patient diagnosed with pneumonia due to SARS-CoV-2. A 20-year-old female patient, primigravid, 36 weeks of gestation, without comorbidities, and diagnosed with severe pneumonia due to the SARS-CoV-2, showed rapid lung deterioration for which their pregnancy was interrupted by surgery. The patient was hospitalized in the Intensive Care Unit (ICU), connected to mechanical ventilation and receiving corticosteroids and antibiotics. The diagnosis of pulmonary fungal infection was made through bronchoalveolar lavage (BAL) culture, and the species identification was performed by sequencing of ß-tubulin. Phylogenetic analysis with related species was performed for the confirmation of species identification. Antifungal susceptibility tests were performed for itraconazole (4 µg/mL), voriconazole (2 µg/mL), and amphotericin B (2 µg/mL). The patient was successfully treated with itraconazole. This is the second worldwide report of pulmonary infection by P. digitatum and the first in Chile. Although it is a fungus that rarely infects humans, it could represent an emerging opportunistic fungal pathogen, with associated risk factors that should be considered in the differential diagnosis of Penicillium species isolated from infections in humans.

[Invasive ocular fungal infection by Scedosporium apiospermum in an immunocompromised patient]. / Infección ocular invasora por Scedosporium apioespermum en un paciente inmunocomprometido.

We report a case of invasive fungal infection with necrotizing conjunctivitis, scleritis and unilateral panuveitis caused by Scedosporium apiospermum in a 78-year-old woman that developed neutropenia by drugs indicated for rheumatoid arthritis. The etiological diagnosis was confirmed by mycological culture of an ocular secretion with the support of MALDI-TOF-TOF analysis and histopathological findings. The treatment involved surgical debridements together with topical solution and systemic therapy with voriconazole and steroids with a favorable evolution after 2 months of treatment. A relapse required a second therapeutic course for an additional 12 months with improvement and eradication of the agent. Fungal conjunctivitis due to S. apiospermum is a rare event associated with immunosuppressed patients. Its treatment involves surgical debridements and prolonged antifungal therapy.

Infección ocular invasora por Scedosporium apioespermum en un paciente inmunocomprometido / Invasive ocular fungal infection by Scedosporium apiospermum in an immunocompromised patient

Resumen Presentamos el caso clínico de una infección fúngica invasora con una conjuntivitis necrosante, escleritis y panuveitis unilateral por Scedosporium apiospermum en una mujer de 78 años con artritis reumatoidea con neutropenia secundaria a fármacos. El diagnóstico etiológico fue confirmado por cultivo micológico de secreción ocular con apoyo de MALDI-TOF-TOF e histopatología. El tratamiento incluyó aseos quirúrgicos asociado a terapia tópica y sistémica con voriconazol y corticoesteroides, con una evolución favorable a los dos meses de tratamiento. Una recaída obligó a un segundo curso terapéutico por 12 meses adicionales con mejoría y erradicación del agente. La conjuntivitis fúngica por S. apiospermum es un evento infrecuente asociado a pacientes inmunocomprometidos. Su tratamiento involucra desbridamientos quirúrgicos y terapia antifúngica prolongada.

Abstract We report a case of invasive fungal infection with necrotizing conjunctivitis, scleritis and unilateral panuveitis caused by Scedosporium apiospermum in a 78-year-old woman that developed neutropenia by drugs indicated for rheumatoid arthritis. The etiological diagnosis was confirmed by mycological culture of an ocular secretion with the support of MALDI-TOF-TOF analysis and histopathological findings. The treatment involved surgical debridements together with topical solution and systemic therapy with voriconazole and steroids with a favorable evolution after 2 months of treatment. A relapse required a second therapeutic course for an additional 12 months with improvement and eradication of the agent. Fungal conjunctivitis due to S. apiospermum is a rare event associated with immunosuppressed patients. Its treatment involves surgical debridements and prolonged antifungal therapy.

[Antifungal activity of mouthwashes against Candida albicans and Rhodotorula mucilaginosa: An in vitro study]. / Actividad antifúngica de los enjuagues bucales frente a Candida albicans y Rhodotorula mucilaginosa: un estudio in vitro.

BACKGROUND: Candida albicans and Rhodotorula mucilaginosa are yeasts of clinical importance in the oral cavity. In immunocompromised patients they can cause some pathologies that must be controlled with antimicrobials. AIMS: To evaluate and compare the antimicrobial efficacy of commercially available mouthrinses against strains of C. albicans and R. mucilaginosa. METHODS: The six mouthwashes studied in vitro were formulated (alone or in combination) with chlorhexidine (CHX) 0.12%, CHX 0.1%, CHX 0.05%, cetylpyridinium chloride (CPC) 0.075%, CPC 0.05%, and essential oils. Ten C. albicans and R. mucilaginosa isolates each were studied. The agar diffusion method (Mueller Hinton II), with incubation at 32°C was used to evaluate the antifungal activity. RESULTS: The results of this study indicate that mouthwashes with CHX 0.1%, CHX 0.12%, CHX 0.05% + CPC 0.05%, CHX 0.12% + CPC 0.05% and CPC 0.075% have an antifungal effect against C. albicans and R. mucilaginosa. CHX 0.1% led to the broadest inhibition zone for C. albicans and R. mucilaginosa (25.65±2.39mm and 40.05±3.31mm). Essential oils did not show any antifungal activity. Statistical analysis showed no statistical difference between mouth rinses CHX 0.1%, CHX 0.12% and CHX 0.12% + CPC 0.05% (p=0.0001) against C. albicans and R. mucilaginosa. CONCLUSIONS: Mouthwashes with CHX showed higher antifungal activity against C. albicans and R. mucilaginosa than other mouthwashes studied.

Actividad antifúngica de los enjuagues bucales frente a Candida albicans y Rhodotorula mucilaginosa: un estudio in vitro / Antifungal activity of mouthwashes against Candida albicans and Rhodotorula mucilaginosa: an in vitro study

ANTECEDENTES: Candida albicans y Rhodotorula mucilaginosa son levaduras de importancia clínica en la cavidad oral. En pacientes inmunocomprometidos estas levaduras pueden producir enfermedades que deben ser controladas con antimicrobianos. OBJETIVOS: Evaluar y comparar la eficacia antifúngica de seis enjuagues bucales comerciales frente a aislamientos de C. albicans y R. mucilaginosa. MÉTODOS: Fue evaluada in vitro la eficacia de seis enjuagues bucales formulados (solos o en combinación) con clorhexidina (CHX) 0,12%; CHX 0,1%; CHX 0,05%; cloruro de cetilpiridinio (CPC) 0,075%; CPC 0,05% y aceites esenciales. Fueron utilizados diez aislamientos de C. albicans y otros diez de R. mucilaginosa. Mediante el método de difusión en placa con agar Mueller Hinton (modificado) se midieron los halos de inhibición previa incubación a 32°C. RESULTADOS: Los resultados de este estudio indican que enjuagues bucales con CHX 0,1%; CHX 0,12%; CHX 0,05% + CPC 0,05%; CHX 0,12% + CPC 0,05% y CPC 0,075% ejercen un efecto antifúngico frente a C. albicans y R. mucilaginosa. CHX 0,1% dio lugar a la mayor zona de inhibición para C. albicans y R. mucilaginosa (25,65±2,39mm y 40,05±3,31mm). El enjuague con aceites esenciales no tuvo actividad antifúngica alguna. El análisis estadístico no mostró diferencia entre los enjuagues bucales CHX 0,1%; CHX 0,12% y CHX 0,12% + CPC 0,05% (p = 0,0001) frente a C. albicans y R. mucilaginosa. CONCLUSIONES: Los enjuagues bucales con CHX mostraron una mejor actividad antifúngica contra C. albicans y R. mucilaginosa que los restantes enjuagues estudiados

BACKGROUND: Candida albicans and Rhodotorula mucilaginosa are yeasts of clinical importance in the oral cavity. In immunocompromised patients they can cause some pathologies that must be controlled with antimicrobials. AIMS: To evaluate and compare the antimicrobial efficacy of commercially available mouthrinses against strains of C. albicans and R. mucilaginosa. METHODS: The six mouthwashes studied in vitro were formulated (alone or in combination) with chlorhexidine (CHX) 0.12%, CHX 0.1%, CHX 0.05%, cetylpyridinium chloride (CPC) 0.075%, CPC 0.05%, and essential oils. Ten C. albicans and R. mucilaginosa isolates each were studied. The agar diffusion method (Mueller Hinton II), with incubation at 32°C was used to evaluate the antifungal activity. RESULTS: The results of this study indicate that mouthwashes with CHX 0.1%, CHX 0.12%, CHX 0.05% + CPC 0.05%, CHX 0.12% + CPC 0.05% and CPC 0.075% have an antifungal effect against C. albicans and R. mucilaginosa. CHX 0.1% led to the broadest inhibition zone for C. albicans and R. mucilaginosa (25.65±2.39mm and 40.05±3.31mm). Essential oils did not show any antifungal activity. Statistical analysis showed no statistical difference between mouth rinses CHX 0.1%, CHX 0.12% and CHX 0.12% + CPC 0.05% (p = 0.0001) against C. albicans and R. mucilaginosa. CONCLUSIONS: Mouthwashes with CHX showed higher antifungal activity against C. albicans and R. mucilaginosa than other mouthwashes studied

Primer reporte de Cryptococcus gattii en Ecuador / First report of Cryptococcus gattii in Ecuador

El presente es el reporte de un caso de un paciente masculino de 37 años, con manifestaciones clínicas de aproximadamente 1 año de evolución caracterizadas por cefalea holocraneana de gran intensidad, fotofobia, diplopía, que se exacerban en los últimos tres meses con alucinaciones, pérdida de audición de oído izquierdo, refirió exposición de 4 años a heces de aves (palomas) por su trabajo en el puerto marítimo.Acudió por consulta privada a valoración y manejo, se le efectúa el diagnóstico de diabetes mellitus tipo 2, por lo cual se prescribió metformina oral, una vez iniciado el tratamiento presentó exacerbación de los síntomas neurológicos, por lo que se indicó internación hospitalaria y se sometió a investigación para diagnóstico de su enfermedad, con presunción de meningitis bacteriana, se le realizó punción lumbar para estudios microbiológicos y se inició tratamiento empírico con glucopeptido y carbapenemico. Con el esquema antimicrobiano presentó leve mejoría en la intensidad de sus síntomas, en el estudio microbiológico del líquido cefalorraquídeo se observaron estructuras micóticas compatibles con Cryptococcus, por lo cual se realizó una segunda punción lumbar para conseguir una muestra adecuada para cultivo y estudio con biología molecular, a fin de establecer el diagnóstico definitivo del microorganismo.

The present is a report of a 37-year-old male patient, who present clinical manifestations of approximately 1 year, by high-intensity holocranial headache, photophobia, diplopia, which were exacerbated in the last three months with hallucinations, loss of Hearing from the left ear, hereported exposure of 4 years to feces of birds (pigeons) for his work in the seaport.The patient was refer to a private consultation to assessment and management, he was diagnosed with type 2 diabetes mellitus, for which oral metformin was prescribed, once the treatment had begun, exacerbation of the neurological symptoms, so hospitalization was indicated, and He underwent research to diagnose his disease, with presumed bacterial meningitis, he underwent lumbar puncture for microbiological studies and began empirical treatment with glucopeptide and carbapenemic; With the antimicrobial scheme, there was a slight improvement in the intensity of its symptoms. In the microbiological study of the preliminary cerebrospinal fluid, fungal structures compatible with Cryptococcus were observe; a second lumbar puncture was performed to obtain an adequate sample for culture and study with molecular biology, in order to establish the definitive diagnosis of the microorganism.

Epidemiología y susceptibilidad antifúngica de especies causantes de candidemia en la ciudad de Valdivia, Chile / Epidemiology and antifungal susceptibility of species producing candidemia in Valdivia, Chile

Resumen Introducción: La epidemiología de la candidemia en Chile es poco conocida, a pesar de ser una enfermedad frecuente y grave. Objetivos: Analizar la incidencia y perfil de susceptibilidad in vitro de especies del género Candida causantes de candidemia en el Hospital Base Valdivia. Material y Métodos: Se identificó todos los casos de candidemia ocurridos en la institución entre marzo de 2009 y agosto de 2011. Se estudió la distribución de especies y susceptibilidad in vitro por el método de disco difusión. Resultados: Se estudiaron 27 episodios de candidemia. La incidencia varió de 0,3 a 0,7 por 1.000 egresos (según servicio clínico). Fueron factores de riesgo: la hospitalización, uso previo de antimicrobianos, edad avanzada y enfermedad de base (insuficiencia renal, enfermedad cardiaca y pulmonar). La especie más frecuente fue C. albicans seguida de C. tropicalis, C. glabrata y C. krusei. Voriconazol fue el antifúngico con mejor actividad in vitro y hubo resultados variables para anfotericina B, fluconazol e itraconazol. Discusión: La incidencia encontrada fue superior a la de países desarrollados. Candida albicans, constituye la especie más frecuente, con alta sensibilidad in vitro a fluconazol, debiendo ser vigiladas las especies de Candida no albicans. Es fundamental desarrollar nuevas investigaciones para detectar potenciales variaciones epidemiológicas.

Background: Candidemia disease in Chile is not very known, despite being a very common and serious disease. Aim: To analyze incidence and susceptibility profile of species from the genus Candida in Hospital Base Valdivia. Material and Methods: All candidemia cases presented between March 2009 and August 2011 in our institution were identified. Species distribution and susceptibility were studied through the disk diffusion method. Results: Twenty-seven candidemia episodes were studied. Incidence varied from 0.3 to 0.7 by a thousand discharges (according to clinical service). Risk factors were hospitalization, preliminary use of antibiotics, aged people and underlying disease (renal failure, heart and lung disease). The most frequent species was C. albicans, followed by C. tropicalis, C. glabrata and C. krusei. Voriconazole was the antifungal showing the best in vitro performance. Amphotericin B, fluconazole and itraconazole presented variable results. Discussion: Incidence found was superior to that described in developed countries. Candida albicans constitute the most frequent species, with high sensitivity levels to fluconazole; Candida no albicans species must be monitored. Developing new research on the topic appears as fundamental to detect potential epidemiological variations.

[Epidemiology and antifungal susceptibility of species producing candidemia in Valdivia, Chile]. / Epidemiología y susceptibilidad antifúngica de especies causantes de candidemia en la ciudad de Valdivia, Chile.

BACKGROUND: Candidemia disease in Chile is not very known, despite being a very common and serious disease. AIM: To analyze incidence and susceptibility profile of species from the genus Candida in Hospital Base Valdivia. MATERIAL AND METHODS: All candidemia cases presented between March 2009 and August 2011 in our institution were identified. Species distribution and susceptibility were studied through the disk diffusion method. RESULTS: Twenty-seven candidemia episodes were studied. Incidence varied from 0.3 to 0.7 by a thousand discharges (according to clinical service). Risk factors were hospitalization, preliminary use of antibiotics, aged people and underlying disease (renal failure, heart and lung disease). The most frequent species was C. albicans, followed by C. tropicalis, C. glabrata and C. krusei. Voriconazole was the antifungal showing the best in vitro performance. Amphotericin B, fluconazole and itraconazole presented variable results. DISCUSSION: Incidence found was superior to that described in developed countries. Candida albicans constitute the most frequent species, with high sensitivity levels to fluconazole; Candida no albicans species must be monitored. Developing new research on the topic appears as fundamental to detect potential epidemiological variations.

Hongos productores de micotoxinas asociados a hierbas medicinales en la Ciudad de São Paulo, Brasil / Mould and mycotoxins associated in medicinal plants in São Paulo, Brazil

Antecedentes: la fitoterapia es una de las más antiguas prácticas utilizadas por la humanidad. Hasta mediados del siglo XIX, cuando se introdujeron los medicamentos, la formulación de estos generalmente era basada en plantas medicinales. Objetivos: Determinar la micobiota y los niveles de aflatoxinas originadas de Aspergillus sección Flavi aislados de las 50 muestras de medicamentos fitoterápicos comercializados actualmente en la ciudad de São Paulo, Brasil. Métodos: Cincuenta (50) muestras de medicamentos fitoterápicos en la forma de hojas (té-25) y cápsulas (25) fueron colectadas de agosto de 2000 a julio de 2001. Los hongos filamentosos aislados fueron identificados al nivel de género de acuerdo con las características morfológicas y criterios taxonómicos. El análisis de aflatoxinas fue realizada por cromatografía de capa fina (TLC). Resultados: El análisis microbiológico mostró que 41 (82 por ciento) de los medicamentos fitoterápicos presentaron un crecimiento fúngico sobre las 100 UFC/g. Un total de 106 especies de seis diferentes géneros fueron aislados (Aspergillus, Penicillium, Mucor, Rhizopus y Alternaria). El género Aspergillus fue el predominante (60.5 por ciento) seguido por Penicillium (20,0 por ciento). Aspergillus niger (30 por ciento) A. flavus (22 por ciento), A. fumigatus (6,5 por ciento) y A. parasiticus fueron las especies de Aspergillus identificadas. Se observó que 13 (56,5 por ciento), de los 23 A. flavus aislados y dos aislados de A. parasiticus produjeron aflatoxinas. Conclusiones: La contaminación observada en la mayoría de los productos y el alto nivel de cepas productoras de aflatoxinas justifica un análisis más cuidadoso de los medicamentos fitoterápicos comercializados y la aplicación de leyes más rigurosas son necesarias para garantizar la calidad de los productos.

Background: phytotherapy is one of the most ancient practices used by humanity. In Antiquity until the middle of the XIX century, when chemotherapeutic drugs were introduced, formulation of medicines was usually based on medicinal plants. Objective: To determine mycobiota and levels of Aspergillus section Flavi aflatoxins isolated from 50 samples of phytotherapeutic remedies currently commercialized in São Paulo, Brazil. Methods: Fifty (50) samples of phytotherapeutic remedies in the form of leaves (teas-25) and powders (capsules-25) were collected from August 2000 to July 2001. Filamentous fungi isolates were identified at the genera level in accordance with morphological characteristics and taxonomic criteria. Aflatoxins were performed by Thin-layer chromatography (TLC). Results: The microbiological analysis showed that 41 (82 percent) of phytotherapeutic remedies presented a fungal growth over 100 CFU/g. A total of 106 species of six different genera were isolated (Aspergillus, Penicillium, Mucor, Rhizopus and Alternaria). The genus Aspergillus was the predominant (60.5 percent) followed by Penicillium genus (20.0 percent). Aspergillus niger (30 percent) A. flavus (22 percent), A. fumigatus (6.5 percent) and A. parasiticus were the species of Aspergillus identified. It was observed that 13 (56.5 percent) of 23 A. flavus isolates and two A. parasiticus isolates produced aflatoxins. Conclusions: The contamination observed in most products and the high level of aflatoxigenic strains justify the concern regarding the execution of more careful analyzes of the commercialized phytotherapeutic remedies and the application of more rigorous laws that may warrant the quality of these products.

Detección de virus parainfluenza en pacientes inmunocomprometidos e inmunocompetentes atendidos en un hospital terciario de la Ciudad de São Paulo, Brasil / Detection of parainfluenza virus in immunocompromised and immunocompetent patients in a tertiary hospital in Sao Paulo, Brazil

Parainfluenza virus infections (PIV) were evaluated in patients with mild and severe infections through real time PCR. One thousand and sixty-seven samples were collected from subjects as follows: 233 adult renal transplanted outpatients, 129 children with congenital heart disease, 381 with adult hematopoietic stem cell patients and 324 hospitalized patients suspected of influenza A (H1N1) pdm09 infection. PIV was detected in 74 (6.9%) samples. VPI-3 was the most frequent (60.8%) and a higher risk was observed for older adults (p = 0.018) and for those who were hematopoietic stem cell transplanted. Further studies are needed to understand the VPI role in patients' at risk for developing serious illness.

Se evaluó la infección por virus parainfluenza (VPI) en pacientes con infecciones leves y graves mediante RPC en tiempo real. Se analizó un total de 1.067 muestras: 233 provenían de pacientes ambulatorios adultos receptores de trasplantes renales, 129 de niños con cardiopatía congénita, 381 de pacientes receptores de trasplantes de precursores hematopoyéticos adultos y 324 de pacientes hospitalizados con sospecha de influenza A (H1N1) pdm09. Se detectó VPI en 74 muestras (6,9%). Siendo VPI-3 el virus más frecuente (60,8%), se observó un mayor riesgo para los adultos mayores (p = 0,018) y para aquellos que fueron receptores de precursores hematopoyéticos. Son necesarios estudios adicionales para entender el papel del VPI en pacientes de riesgo para desarrollar enfermedad grave.

[Detection of parainfluenza virus in immunocompromised and immunocompetent patients in a tertiary hospital in Sao Paulo, Brazil]. / Detección de virus parainfluenza en pacientes inmunocomprometidos e inmunocompetentes atendidos en un hospital terciario de la Ciudad de São Paulo, Brasil.

Parainfluenza virus infections (PIV) were evaluated in patients with mild and severe infections through real time PCR. One thousand and sixty-seven samples were collected from subjects as follows: 233 adult renal transplanted outpatients, 129 children with congenital heart disease, 381 with adult hematopoietic stem cell patients and 324 hospitalized patients suspected of influenza A (H1N1) pdm09 infection. PIV was detected in 74 (6.9%) samples. VPI-3 was the most frequent (60.8%) and a higher risk was observed for older adults (p = 0.018) and for those who were hematopoietic stem cell transplanted. Further studies are needed to understand the VPI role in patients' at risk for developing serious illness.

Un caso de queratitis micótica por Fusarium solani en Valdivia, Chile / A case of mycotic keratitis due to Fusarium solani in Valdivia, Chile

Antecedentes: La queratitis micótica es una de las principales infecciones oculares. Se necesita un tratamiento específico que depende de la naturaleza del microorganismo infectante. Su pronóstico suele ser grave y depende de un diagnóstico precoz y una terapia adecuada. Caso clínico: Se describe un caso clínico de queratitis por Fusarium solani en un paciente procedente de un área rural, afectado entre mayo y octubre de 2011 por un traumatismo corneal causado por partículas de polvo en Valdivia, Chile. El examen directo del raspado corneal reveló, en dos ocasiones, la presencia de abundantes hifas septadas. Todos los cultivos fueron positivos para el mismo hongo, que fue identificado como Fusarium solani mediante técnicas fenotípicas y secuenciación de los genes nucleares ribosomales. El paciente fue tratado inicialmente con anfotericina B, y después, con voriconazol oral e intravenoso, tratamiento al que respondió exitosamente, aunque la opacidad corneal persistió. Conclusiones: Aun cuando las queratomicosis en Chile son poco frecuentes, deben tomarse en consideración sus particularidades diagnósticas para poder instaurar el tratamiento más eficaz. Así, se hace indispensable la rápida visualización del hongo en la lesión, un eficiente aislamiento del agente etiológico en cultivo y una rápida identificación, lo que para el caso particular de las especies del género Fusarium requiere el empleo de técnicas moleculares de secuenciación (AU)

Background: Keratomycosis is one of the most prevalent ophthalmic infections, which needs a specific treatment depending on the nature of the infecting fungus. The prognosis is usually severe and depends on an early diagnosis and suitable therapy. Case report: We describe a case of keratitis due to Fusarium solani in a patient from a rural area, who, between May and October 2011, suffered a corneal trauma caused by dust particles in Valdivia, Chile. On two occasions, direct examination of eye scrapes revealed abundant septate hyphae. All cultures were positive for the same fungus, which was identified as Fusarium solani by phenotypic characterization and sequencing of ribosomal nuclear genes. The patient was initially treated with amphotericin B and afterwards successfully responded to a treatment with oral and intravenous voriconazole, although corneal opacity persisted. Conclusions: Although keratomycosis in Chile is rare, its diagnostic particularities must be taken into consideration to establish the most effective treatment. Thus, a rapid visualization of the fungus in the lesion, an efficient isolation of the etiologic agent in pure culture is essential, as well as its rapid identification, which requires the use of molecular sequencing techniques in the case of Fusarium species (AU)

[A case of mycotic keratitis due to Fusarium solani in Valdivia, Chile]. / Un caso de queratitis micótica por Fusarium solani en Valdivia, Chile.

BACKGROUND: Keratomycosis is one of the most prevalent ophthalmic infections, which needs a specific treatment depending on the nature of the infecting fungus. The prognosis is usually severe and depends on an early diagnosis and suitable therapy. CASE REPORT: We describe a case of keratitis due to Fusarium solani in a patient from a rural area, who, between May and October 2011, suffered a corneal trauma caused by dust particles in Valdivia, Chile. On two occasions, direct examination of eye scrapes revealed abundant septate hyphae. All cultures were positive for the same fungus, which was identified as Fusarium solani by phenotypic characterization and sequencing of ribosomal nuclear genes. The patient was initially treated with amphotericin B and afterwards successfully responded to a treatment with oral and intravenous voriconazole, although corneal opacity persisted. CONCLUSIONS: Although keratomycosis in Chile is rare, its diagnostic particularities must be taken into consideration to establish the most effective treatment. Thus, a rapid visualization of the fungus in the lesion, an efficient isolation of the etiologic agent in pure culture is essential, as well as its rapid identification, which requires the use of molecular sequencing techniques in the case of Fusarium species.

Yeasts isolated from nosocomial urinary infections: antifungal susceptibility and biofilm production / Levaduras aisladas de pacientes con infecciones urinarias nosocomiales: sensibilidad antifúngica y producción de biofilm

Background. Urinary Candida infections in the hospital environment are frequent and need to be better understood. Aims. To compare the results of antifungal susceptibility profiles of yeasts isolated from patients with urinary infections obtained by broth microdilution method (BM) and by disk diffusion (DD), and also evaluate the capacity of these yeasts to form biofilms. Methods. Only yeasts obtained from pure urine cultures with counts higher than 105 colony-forming units per milliliter, without bacteria development, of symptomatic patients were included. The isolates were identified by classical methods and the antifungal susceptibility tests were performed with the following drugs: amphotericin B, ketoconazole, fluconazole, itraconazole, voriconazole and caspofungin. The biofilm studies were carried out in polystyrene microtitration plates. Results. Ninety-five yeasts isolates were analyzed, including 40 Candida albicans, 31 Candida glabrata, 24 Candida tropicalis. In general, the majority of the isolates were susceptible to the tested drugs but some resistance was observed, especially against fluconazole. Great variability in the antifungal susceptibility results was observed with the different tested drugs and a few discrepancies were observed between both methods. We suggest that in case of DD resistance this result should be confirmed by BM, the standard method. C. tropicalis isolates showed high biofilm production (91.7%) compared to C. albicans (82.5%) and C. glabrata (61.3%), with statistical significance (p = 0.0129). Conclusions. Candiduria in critical patients requires major attention and a better control. The different susceptibility results obtained in this study showed the need to identify yeasts up to the species level, especially in patients with urinary tract infection. The development of techniques of antifungal susceptibility tests can help the clinicians in the empiric treatment of candiduria (AU)

Antecedentes. Las infecciones urinarias producidas por especies del género Candida en el ámbito hospitalario son frecuentes, por lo que se requieren mayores conocimientos. Objetivos. Examinar los resultados de los perfiles de sensibilidad de las levaduras aisladas de pacientes con infección urinaria a los fármacos antimicóticos, comparar los resultados obtenidos con las técnicas de microdilución en caldo y difusión en agar con disco, y valorar la capacidad de estas levaduras para producir biofilm. Métodos. Solo se incluyeron en el estudio las levaduras obtenidas a partir de urocultivos puros de pacientes sintomáticos con recuentos superiores a 105 unidades formadoras de colonias, sin el desarrollo de bacterias. Las levaduras se identificaron con técnicas clásicas y se realizaron pruebas de sensibilidad frente a los antimicóticos siguientes: anfotericina B, ketoconazol, fluconazol, itraconazol, voriconazol y caspofungina. Los exámenes de producción de biofilm se efectuaron en placas de microtitulación de poliestireno. Resultados. Se analizaron 95 aislamientos de levaduras que incluían 40 Candida albicans, 31 Candida glabrata y 24 Candida tropicalis. En general, la mayoría de los aislamientos eran sensibles a los fármacos examinados, aunque se observaron algunas resistencias, en especial al fluconazol. Se observó una variabilidad considerable en los resultados de la sensibilidad a los diferentes antimicóticos examinados, detectándose algunas discrepancias entre ambos métodos de examen. Sugerimos que los casos valorados como resistentes por difusión con disco se confirmen mediante microdilución en caldo, que es el método de referencia. Los aislamientos de C. tropicalis mostraron una elevada producción de biofilm (91,7%) en comparación con C. albicans (82,5%) y C. glabrata (61,3%), siendo la diferencia estadísticamente significativa (p = 0,0129). Conclusiones. Es preciso prestar mayor atención a la candiduria detectada en pacientes en estado crítico, al igual que un mejor control. Los diferentes resultados de sensibilidad a los antimicóticos obtenidos en el presente estudio demuestran la necesidad de identificar las especies de las levaduras aisladas de pacientes con infecciones del tracto urinario. El progreso de las técnicas de sensibilidad a los antimicóticos puede ayudar a los médicos en el tratamiento empírico de la candiduria (AU)

Yeasts isolated from nosocomial urinary infections: antifungal susceptibility and biofilm production.

BACKGROUND: Urinary Candida infections in the hospital environment are frequent and need to be better understood. AIMS: To compare the results of antifungal susceptibility profiles of yeasts isolated from patients with urinary infections obtained by broth microdilution method (BM) and by disk diffusion (DD), and also evaluate the capacity of these yeasts to form biofilms. METHODS: Only yeasts obtained from pure urine cultures with counts higher than 10(5) colony-forming units per milliliter, without bacteria development, of symptomatic patients were included. The isolates were identified by classical methods and the antifungal susceptibility tests were performed with the following drugs: amphotericin B, ketoconazole, fluconazole, itraconazole, voriconazole and caspofungin. The biofilm studies were carried out in polystyrene microtitration plates. RESULTS: Ninety-five yeasts isolates were analyzed, including 40 Candida albicans, 31 Candida glabrata, 24 Candida tropicalis. In general, the majority of the isolates were susceptible to the tested drugs but some resistance was observed, especially against fluconazole. Great variability in the antifungal susceptibility results was observed with the different tested drugs and a few discrepancies were observed between both methods. We suggest that in case of DD resistance this result should be confirmed by BM, the standard method. C. tropicalis isolates showed high biofilm production (91.7%) compared to C. albicans (82.5%) and C. glabrata (61.3%), with statistical significance (p=0.0129). CONCLUSIONS: Candiduria in critical patients requires major attention and a better control. The different susceptibility results obtained in this study showed the need to identify yeasts up to the species level, especially in patients with urinary tract infection. The development of techniques of antifungal susceptibility tests can help the clinicians in the empiric treatment of candiduria.

The water supply system as a potential source of fungal infection in paediatric haematopoietic stem cell units.

BACKGROUND: We conducted a prospective study to investigate the presence of microfungal contamination in the water supply system of the Oncology Paediatric Institute, São Paulo-Brazil after the occurrence of one invasive Fusarium solani infection in a patient after Haematopoietic Stem Cell Transplantation (HSCT). During a twelve-month period, we investigated the water supply system of the HSCT unit by monitoring a total of fourteen different collection sites. METHODS: One litre of water was collected in each location, filtered through a 0.45 µm membrane and cultured on SDA to detect the presence of filamentous fungi. Physicochemical analyses of samples were performed to evaluate the temperature, turbidity, pH, and the concentration of free residual chlorine. RESULTS: Over the 12 months of the study, 164 samples were collected from the water supply system of the HSCT unit, and 139 of the samples tested positive for filamentous fungi (84.8%), generating a total of 2,362 colonies. Cladosporium spp., Penicillium spp., Purpureocillium spp. and Aspergillus spp. were ranked as the most commonly found genera of mould in the collected samples. Of note, Fusarium solani complex isolates were obtained from 14 out of the 106 samples that were collected from tap water (mean of 20 CFU/L). There was a positive correlation between the total number of fungal CFU obtained in all cultures and both water turbidity and temperature parameters. Our findings emphasise the need for the establishment of strict measures to limit the exposure of high-risk patients to waterborne fungal propagules. CONCLUSIONS: We were able to isolate a wide variety of filamentous fungi from the water of the HSCT unit where several immunocompromised patients are assisted.

Molecular phylogeny and phenotypic variability of clinical and environmental strains of Aspergillus flavus.

Aspergillus flavus is the second most common cause of aspergillosis infection in immunocompromised patients and is responsible for the production of aflatoxins. Little is known about the population structure of A. flavus, although recent molecular and phenotypic data seem to demonstrate that different genetic lineages exist within this species. The aim of this study was to carry out a morphological, physiological, and molecular analysis of a set of clinical and environmental isolates to determine whether this variability is due to species divergence or intraspecific diversity, and to assess whether the clinical isolates form a separate group. The amdS and omtA genes were more phylogenetically informative than the other tested genes and their combined analysis inferred three main clades, with no clear distinction between clinical and environmental isolates. No important morphological and physiological differences were found between the members of the different clades, with the exception of the assimilation of d-glucosamine, which differentiates the members of the clade II from the others.

Aspergillus novoparasiticus: a new clinical species of the section Flavi.

During a survey on the incidence of Aspergillus in clinical environments, we found some interesting isolates that were morphologically similar to Aspergillus parasiticus, but differed in the color of the colonies and in the pattern of their conidial ornamentation. In the present study, those isolates were characterized using a polyphasic approach. A phylogenetic analysis was carried out, based on partial fragments of the acetamidase (amdS) and O-methyltransferase (omtS) genes and the internal transcribed spacer (ITS) region of rDNA. This information was combined with a detailed morphological and physiological study that included aflatoxin production and assimilation profiles of different carbon and nitrogen sources. The phenotypic and genotypic results support the proposal of a new species, Aspergillus novoparasiticus, phylogenetically placed in a distinct sister clade to that of A. parasiticus. The former has lobate-reticulate conidia and does not produce aspergillic acid on AFPA or organic acids on CREA, while A. parasiticus has echinulate conidia and produces aspergillic and organic acids. In addition, this new species, as well as A. parasiticus, produces aflatoxins B1, B2, G1 and G2.

Onychomycosis in São Paulo, Brazil.

Onychomycosis is a fungal infection of the nails with broad aetiological scope, and it represents 18-40% of all onychopathies and 39% of all superficial mycotic infections. From July 1996 to December 1999, samples of nails were collected from 588 patients with presumptive diagnosis of onychomycosis at the Dermatology and Mycology Divisions EPM\UNIFESP, Brazil, and the diagnosis was confirmed in 247 of these cases. The most common pathogens isolated in this study were yeasts in 52% of positive cultures (Candida albicans 18.3%, Candida parapsilosis 13.8%, other species of Candida 15.4% and other yeasts 4.6%), followed by dermatophytes in 40.6% of positive cultures (the most commonly isolated organisms were Trichophyton rubrum in 33.2%, followed by Trichophyton mentagrophytes in 6.3% and others 1.2%). Non-dermatophyte moulds were isolated in 7.4% of positive cultures (Fusarium spp. 4.5%, Nattrassia mangiferae 2.3% and Aspergillus spp. 0.6%). Distal and lateral subungual onychomycosis (DLSO) was the commonest clinical pattern 44.6% followed by free edge onycholysis (FEO) 38.8% and others. In conclusion, this study demonstrated that T. rubrum is the main agent causing onychomycosis in toenails, and species of genus Candida were the main agents isolated in fingernail onychomycosis in our region.

Performance of the Albicans ID2omicron chromogenic medium for rapid identification of Candida albicans

The aim of our study was to evaluate the accuracy of the chromogenic media Albicans ID2omicron (bioMérieux, France) for the identification of Candida albicans among 330 yeast strains. All C. albicans (100) and C. dubliniensis (20) strains exhibited blue color when cultured on Albicans ID2omicron. However, the blue color was also exhibited by cultures of C. rugosa (30/30) and C. tropicalis (3/50) isolates.

O objetivo do nosso estudo foi avaliar a eficácia do meio cromogênico Albicans ID2ômicron (bioMérieux, France) na identificação de Candida albicans entre 330 amostras de leveduras. As cepas de C. albicans (100) e C. dubliniensis (20) exibiram coloração azul quando semeadas em Albicans ID2ômicron. Contudo, a coloração azul também foi verificada em culturas de C. rugosa (30/30) e C. tropicalis (3/50).