Analyses of the genetic diversity and population structures of Histoplasma capsulatum clinical isolates from Mexico, Guatemala, Colombia and Argentina, using a randomly amplified polymorphic DNA-PCR assay.

We studied the genetic diversity and the population structure of human isolates of Histoplasma capsulatum, the causative agent of histoplasmosis, using a randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) assay to identify associations with the geographic distribution of isolates from Mexico, Guatemala, Colombia and Argentina. The RAPD-PCR pattern analyses revealed the genetic diversity by estimating the percentage of polymorphic loci, effective number of alleles, Shannon's index and heterozygosity. Population structure was identified by the index of association (IA) test. Thirty-seven isolates were studied and clustered into three groups by the unweighted pair-group method with arithmetic mean (UPGMA). Group I contained five subgroups based on geographic origin. The consistency of the UPGMA dendrogram was estimated by the cophenetic correlation coefficient (CCCr = 0.94, P = 0.001). Isolates from Mexico and Colombia presented higher genetic diversity than isolates from Argentina. Isolates from Guatemala grouped together with the reference strains from the United States of America and Panama. The IA values suggest the presence of a clonal population structure in the Argentinian H. capsulatum isolates and also validate the presence of recombining populations in the Colombian and Mexican isolates. These data contribute to the knowledge on the molecular epidemiology of histoplasmosis in Latin America.

Brote de histoplasmosis en la Escuela de Cadetes de la Base Aérea de Morón, Provincia de Buenos Aires, República Argentina / Histoplasmosis outbreak in Morón, Buenos Aires Province, Argentina

Se describe un brote de histoplasmosis que afectó a 6 cadetes de la Fuerza Aérea Argentina, sin antecedentes patológicos previos. Todos consultaron por problemas respiratorios después de haber limpiado un hangar. En ese recinto se encontraron abundantes deyecciones de animales, presuntamente de palomas y murciélagos. Los pacientes sufrieron fiebre, mialgias, taquipnea y tos no productiva. Las radiografías y tomografías de tórax mostraron imágenes pulmonares micronodulares, engrosamiento de los tabiques interalveolares y adenopatías hiliares. Todos tuvieron una evolución favorable y no requirieron tratamiento antifúngico. Las pruebas de inmunodifusión y contrainmunoelectroforesis con antígenos de Histoplasma capsulatum fueron positivas, al igual que las intradermorreacciones con histoplasmina. Se recogieron 5 muestras de tierra del lugar, las que fueron inoculadas por vía intraperitoneal a 20 hámsteres. De los cultivos de hígado y bazo de dichos animales se consiguió aislar la fase micelial de H. capsulatum. La cepa aislada se comparó con las obtenidas de 12 pacientes argentinos utilizando perfiles genéticos y se observó un clado único con más de 96% de similitud, lo que confirma la homogeneidad de las cepas argentinas. Si bien la histoplasmosis es endémica en la Pampa húmeda, este es el primer brote totalmente documentado al sur del paralelo 34°.

An histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.

La coccidioidomicosis en Argentina, 1892-2009 / Coccidioidomycosis in Argentina, 1892-2009

Los casos clínicos de coccidioidomicosis en Argentina son pocos y han tenido lugar fundamentalmente en la extensa región árida precordillerana. Este trabajo tiene como objetivos realizar una revisión retrospectiva del total de casos de coccidioidomicosis documentados en Argentina desde el año 1892 hasta 2009 y describir una serie de casos ocurridos en los últimos 4 años. En 117 años se documentaron 128 casos. Desde la primera descripción de la enfermedad en 1892 hasta 1939 se registraron 6 casos; desde 1940 hasta 1999, 59 casos (6-14 casos cada 10 años); y los 63 casos restantes (49% del total histórico) se produjeron en el último decenio. La mediana de edad de los 34 pacientes registrados en el período 2006-2009 fue de 31 años (rango: 7-89), la relación hombre:mujer fue 1,3:1; 12 de estos individuos eran inmunocomprometidos. Veintiséis casos se confirmaron por examen microscópico, por cultivo o por ambos procedimientos; los casos restantes se confirmaron por serología. Todos los aislamientos recuperados fueron identificados como Coccidioides posadasii. Treinta pacientes residían en una amplia área geográfica con epicentro en el valle de Catamarca. Entre 2006 y 2009, la tasa de incidencia en la provincia de Catamarca se incrementó desde valores históricos inferiores a 0,5 casos cada 100 000 habitantes hasta 2,0 casos cada 100 000 habitantes. Este aumento sugiere una emergencia de la coccidioidomicosis en el área.

Clinical cases of coccidioidomycosis are rare in Argentina and are generally found in the large arid precordilleran area of the country. This study aims to perform a retrospective review of all coccidioidomycosis cases documented in the country from 1892 to 2009, and to describe those occurring in the last 4 years. One hundred and twenty eight cases were documented in the 117 year-period. Since the original description of the disease in 1892 until 1939, only 6 cases were registered; between 1940 and 1999, 59 (6-14/10 yrs) and the remaining 63 (49% of total cases) occurred in the last decade. The median age of 34 patients registered in 2006-2009 was 31 years (range: 7-89), male/female ratio was 1.3:1 and 12 patients were immunocompromised. Twenty-six cases were confirmed by direct microscopy and/or culture whereas the remaining ones by serology. All isolates were identified as Coccidioides posadasii. Thirty patients lived in a vast geographic region with epicenter in Catamarca Valley. Between 2006 and 2009, annual disease incidence rates in Catamarca Province increased from historical values below 0.5/100,000 to 2/100,000 inhabitants. Such increase suggests an emergency of coccidioidomycosis in that region.

[Histoplasmosis outbreak in Morón, Buenos Aires Province, Argentina]. / Brote de histoplasmosis en la Escuela de Cadetes de la Base Aérea de Morón, Provincia de Buenos Aires, República Argentina.

A histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.

[Coccidioidomycosis in Argentina, 1892-2009]. / La coccidioidomicosis en Argentina, 1892-2009.

Clinical cases of coccidioidomycosis are rare in Argentina and are generally found in the large arid precordilleran area of the country. This study aims to perform a retrospective review of all coccidioidomycosis cases documented in the country from 1892 to 2009, and to describe those occurring in the last 4 years. One hundred and twenty eight cases were documented in the 117 year-period. Since the original description of the disease in 1892 until 1939, only 6 cases were registered; between 1940 and 1999, 59 (6-14/10 yrs) and the remaining 63 (49% of total cases) occurred in the last decade. The median age of 34 patients registered in 2006-2009 was 31 years (range: 7-89), male/female ratio was 1.3:1 and 12 patients were immunocompromised. Twenty-six cases were confirmed by direct microscopy and/or culture whereas the remaining ones by serology. All isolates were identified as Coccidioides posadasii. Thirty patients lived in a vast geographic region with epicenter in Catamarca Valley. Between 2006 and 2009, annual disease incidence rates in Catamarca Province increased from historical values below 0.5/100,000 to 2/100,000 inhabitants. Such increase suggests an emergency of coccidioidomycosis in that region.

[Molecular diagnosis of human histoplasmosis in whole blood samples]. / Diagnóstico molecular de histoplasmosis humana en muestras de sangre entera.

To assess the value of using whole blood samples for the molecular diagnosis of histoplasmosis, we applied an in-house DNA extraction method and a nested PCR targeting a 210 bp specific segment of the Histoplasma capsulatum HcP100 gene. A whole blood volume of 2.5-3 milliliters was centrifuged and the cellular pellet was treated with Trichoderma harzianum lyticase and proteinase K prior to applying a conventional phenol DNA extraction. This procedure allowed complete cell lysis, high DNA yield and specific amplification. The PCR detection limit was 0.25-1 yeast cells/ml of blood sample. The method was assessed on 31 blood samples from 19 patients with microbiological diagnosis of histoplasmosis, 30 healthy persons and 21 patients with other mycoses or mycobacterial diseases. Positive results were obtained in samples from 17/19 patients with histoplasmosis (14/15 immunocompromised and 3/4 without known immunological disorder). Blood samples from the 30 healthy controls and 20 patients with other conditions proved negative; the only false positive result was obtained from a patient with Mycobacterium avium-intracellulare infection. With 89% sensitivity and 98% specificity, this molecular method for detection of the agent in blood shows promising for the rapid diagnosis of human histoplasmosis.

Diagnóstico molecular de histoplasmosis humana en muestras de sangre entera / Molecular diagnosis of human histoplasmosis in whole blood samples

Se evaluó el uso de sangre entera para el diagnóstico molecular de histoplasmosis utilizando un método artesanal de extracción de ADN fúngico y una PCR anidada que amplifica una porción del gen HcP100 específica de Histoplasma capsulatum. La sangre entera se trató con liticasa, enzima lisante de Trichoderma harzianum y proteinasa K, seguido de una extracción fenólica. Este tratamiento permitió una lisis completa de las células, mostró buen rendimiento en la obtención de ADN y posibilitó la detección de la banda de 210 pb específica de H. capsulatum en la PCR anidada. El límite de detección fue de 0,25-1 levaduras/ml de sangre. El método se evaluó en 31 muestras de sangre de 19 pacientes con diagnóstico microbiológico de histoplasmosis, en 21 muestras de pacientes con otras micosis o infecciones por micobacterias y en 30 controles sanos. La PCR fue positiva en sangre para 17/19 pacientes con histoplasmosis (14/15 inmunocomprometidos y 3/4 sin inmunocompromiso aparente). Las muestras de sangre de los 30 controles sanos y de 20 pacientes con otras patologías fueron negativas, sólo hubo un falso positivo correspondiente a un paciente con infección por Mycobacterium avium-intracellulare. El método presentó 89% de sensibilidad y 96% de especificidad para el diagnóstico de histoplasmosis en sangre entera.

To assess the value of using whole blood samples for the molecular diagnosis of histoplasmosis, we applied an in-house DNA extraction method and a nested PCR targeting a 210 bp specific segment of the Histoplasma capsulatum HcP100 gene. A whole blood volume of 2.5-3 milliliters was centrifuged and the cellular pellet was treated with Trichoderma harzianum lyticase and proteinase K prior to applying a conventional phenol DNA extraction. This procedure allowed complete cell lysis, high DNA yield and specific amplification. The PCR detection limit was 0.25-1 yeast cells/ml of blood sample. The method was assessed on 31 blood samples from 19 patients with microbiological diagnosis of histoplasmosis, 30 healthy persons and 21 patients with other mycoses or mycobacterial diseases. Positive results were obtained in samples from 17/19 patients with histoplasmosis (14/15 immunocompromised and 3/4 without known immunological disorder). Blood samples from the 30 healthy controls and 20 patients with other conditions proved negative; the only false positive result was obtained from a patient with Mycobacterium avium-intracellulare infection. With 89% sensitivity and 98% specificity, this molecular method for detection of the agent in blood shows promising for the rapid diagnosis of human histoplasmosis.

[Epidemiological status of mycoses in the Argentine Republic]. / Situación de las micosis en la República Argentina.

We herein report the results of a retrospective nationwide survey on mycoses diagnosed between January and December, 2004. The study included data provided by 72 laboratories located in 19 provinces and in Buenos Aires City. Out of 801,805 microbiological specimens processed that year, only 62,681 (8%) were submitted to mycological studies. A total of 23,600 mycoses cases were diagnosed: 11,107 (47%) superficial mycoses, 10,830 (46%) mucosal candidiasis and 1,663 (7%) deep mycoses. Relative frequencies of superficial mycoses did not differ significantly (p > 0.05) from frequencies observed in a previous study covering Buenos Aires City and Province (1993), and from two countrywide surveys conducted by the National Network of Mycology Laboratories and National Quality Control Program (NNML and NQCP) in 1999 and 2002. The most frequent deep mycoses were yeast fungaemia (34%), cryptococcosis (20%), broncho-pulmonary aspergillosis (13%), histoplasmosis (11%), paracoccidioidomycosis (7%) and pneumocystosis (5%). In contrast with results of four previous nationwide studies on broncho-pulmonary mycoses including a survey performed by NNML and NQCP in 2002, our study revealed that histoplasmosis prevailed over paracoccidioidomycosis, thus ranking for the first time as the most frequent endemic mycosis in Argentina.

Situación de las micosis en la República Argentina / Epidemiological status of mycoses in the Argentine Republic

Se presentan los resultados de una encuesta nacional sobre micosis diagnosticadas entre enero y diciembre de 2004, con datos provistos por 72 laboratorios de 19 provincias y la Ciudad Autónoma de Buenos Aires. De las 801.805 muestras microbiológicas procesadas ese año, sólo 62.681 (8%) fueron sometidas a estudios micológicos. Se diagnosticaron 23.600 casos de micosis: 11.107 (47%) superficiales, 10.830 (46%) candidiasis de las mucosas y 1.663 (7%) profundas. La frecuencia de agentes de micosis superficiales no sufrió cambios significativos (p>0,05) cuando se comparó con un estudio realizado en población de Provincia y Ciudad de Buenos Aires y dos estudios realizados en 1999 y 2002 por la Red Nacional de Laboratorios y el Programa Nacional de Control de Calidad en Micología (RNLM y PNCCM). Del total de micosis profundas, las más frecuentes fueron fungemia por levaduras (34%), criptococosis (20%), aspergilosis broncopulmonar (13%), histoplasmosis (11%), paracoccidioidomicosis (7%) y neumocistosis (5%). En contraste con los resultados de cuatro estudios previos sobre micosis broncopulmonares, incluyendo el realizado por RNLM y PNCCM en 2002, la histoplasmosis aumentó ubicándose como la micosis endémica más frecuente en Argentina, superando a la paracoccidioidomicosis.

We herein report the results of a retrospective nationwide survey on mycoses diagnosed between January and December, 2004. The study included data provided by 72 laboratories located in 19 provinces and in Buenos Aires City. Out of 801,805 microbiological specimens processed that year, only 62,681 (8%) were submitted to mycological studies. A total of 23,600 mycoses cases were diagnosed: 11,107 (47%) superficial mycoses, 10,830 (46%) mucosal candidiasis and 1,663 (7%) deep mycoses. Relative frequencies of superficial mycoses did not differ significantly (p>0.05) from frequencies observed in a previous study covering Buenos Aires City and Province (1993), and from two countrywide surveys conducted by the National Network of Mycology Laboratories and National Quality Control Program (NNML and NQCP) in 1999 and 2002. The most frequent deep mycoses were yeast fungaemia (34 %), cryptococcosis (20%), broncho-pulmonary aspergillosis (13%), histoplasmosis (11%), paracoccidioidomycosis (7%) and pneumocystosis (5%). In contrast with results of four previous nationwide studies on broncho-pulmonary mycoses including a survey performed by NNML and NQCP in 2002, our study revealed that histoplasmosis prevailed over paracoccidioidomycosis, thus ranking for the first time as the most frequent endemic mycosis in Argentina.

[Disk diffusion method for fluconazole susceptibility testing of Candida spp. isolates]. / Método de difusión con discos para la determinación de sensibilidad a fluconazol en aislamientos de Candida spp.

In order to standardize and evaluate a disk diffusion method with visual reading to detect in vitro fluconazole susceptibility of yeast, 1193 clinical isolates were tested. These included 584 Candida albicans, 196 Candida parapsilosis, 200 Candida tropicalis, 113 Candida glabrata, 50 Candida krusei and 50 Candida spp. and other opportunistic yeasts. The disks were manufactured in the INEI-ANLIS "Dr. Carlos G. Malbrán. The disk diffusion method results were compared to MIC results obtained by the reference CLSI M27-A2 broth microdilution method modified by EUCAST. The interpretative breakpoints for in vitro susceptibility testing of fluconazole were established at: zone diameter > or =16 mm for MIC < or =8 microg/ml (susceptible isolates), between 9 and 15 mm for MIC = 16-32 microg/ml (susceptible dose-dependent isolates), and < or =8 mm for MIC > or =64 microg/ml (resistant isolates). Overall agreement between the two methods was 94.7%, with 0.2% very major errors, and 0.3% major errors. Inter - and intralaboratory agreement was good. The disk diffusion method for drug susceptibility testing of Candida spp. isolates is inexpensive, reliable and reproducible. However, when the inhibition zone diameter is < or =15 mm, it is advisable to test the isolate by the reference microdilution method.

[Histoplasma capsulatum with aberrant morphology isolated in Argentina]. / Aislamientos de Histoplasma capsulatum con morfología aberrante obtenidos en la República Argentina.

For the first time in Argentina, we describe a strain of Histoplasma capsulatum var. capsulatum with an aberrant morphology that was isolated from a single patient with AIDS. Mycelial phase cultures on agar Sabouraud at 25-28 degrees C showed white, glabrous, umbilicated and centrally radiated colonies. Unusual microscopic findings were the absence of typical conidia, the presence of terminal/intercalary chlamydoconidia with a diameter of 4 pm and of thickened hyphae. Fungal identification was confirmed by the detection of bands H and M species specific antigens in mycelial culture supernatants and reversion to the typical yeast phase on agar brain-heart-cysteine at 37 degrees C. The genomic DNA profile obtained by RAPD-PCR with primers 1281-1283 coincided with the predominant profile of H. capsulatum among isolates from Argentine patients.

Método de difusión con discos para la determinación de sensibilidad a fluconazol en aislamientos de Candida spp / Disk diffusion method for fluconazole susceptibility testing of Candida spp. isolates

Se estudiaron 1193 aislamientos clínicos para estandarizar y evaluar un método de difusión con discos de fluconazol de lectura visual, que permita detectar levaduras sensibles al antifúngico. Las especies analizadas fueron: Candida albicans (n=584), Candida parapsilosis (n=196), Candida tropicalis (n=200), Candida glabrata (n=113), Candida krusei (n=50), Candida spp. y otras levaduras oportunistas (n=50). Los discos fueron manufacturados en el INEI-ANLIS "Dr. Carlos G. Malbrán". Se midieron los halos de inhibición del crecimiento producidos por fluconazol y la concentración inhibitoria mínima (CIM) por el método de referencia M27-A2 modificado por EUCAST. Se establecieron los valores de corte del método de difusión en: ≥16 mm para levaduras sensibles a fluconazol (CIM ≤ 8 µg/ml), entre 9 y 15 mm para sensibles dependientes de la dosis (CIM = 16-32 mg/ml) y ≤ 8 mm para resistentes (CIM ≥ 64 µg/ml). El método de difusión tuvo 94,7% de concordancia con el de referencia, con 0,2% de errores very major y 0,3% de errores major. La reproducibilidad inter e intralaboratorio fue muy buena. Para detectar aislamientos sensibles a fluconazol, este método resulta confiable y de bajo costo; sin embargo, es conveniente que los aislamientos con halos ≤ 15 mm sean reevaluados por el método de referencia.

In order to standardize and evaluate a disk diffusion method with visual reading to detect in vitro fluconazole susceptibility of yeast, 1193 clinical isolates were tested. These included 584 Candida albicans, 196 Candida parapsilosis, 200 Candida tropicalis, 113 Candida glabrata, 50 Candida krusei and 50 Candida spp. and other opportunistic yeasts. The disks were manufactured in the INEI-ANLIS "Dr. Carlos G. Malbrán". The disk diffusion method results were compared to MIC results obtained by the reference CLSI M27-A2 broth microdilution method modified by EUCAST. The interpretative breakpoints for in vitro susceptibility testing of fluconazole were established at: zone diameter ≥ 16 mm for MIC ≤ 8 µg/ml (susceptible isolates), between 9 and 15 mm for MIC = 16-32 mg/ml (susceptible dose-dependent isolates), and ≤ 8 mm for MIC ≥ 64 µg/ml (resistant isolates). Overall agreement between the two methods was 94.7%, with 0.2% very major errors, and 0.3% major errors. Inter - and intralaboratory agreement was good. The disk diffusion method for drug susceptibility testing of Candida spp. isolates is inexpensive, reliable and reproducible. However, when the inhibition zone diameter is ≤ 15 mm, it is advisable to test the isolate by the reference microdilution method.

Estudio multicéntrico de fungemias por levaduras en la República Argentina / Multicenter study of bloodstream infections due to yeasts in Argentina

La incidencia de candidemias aumentó aproximadamente en un 500% en hospitales de alta complejidad y se observó un cambio en la distribución de especies del género Candida, con un incremento de las levaduras no Candida albicans. Con el objeto de conocer la distribución de especies asociadas a fungemias por levaduras en Argentina y determinar su sensibilidad a los antifúngicos de uso convencional, se realizó un estudio multicéntrico durante el período abril 1999 a abril 2000. Participaron 36 instituciones del país. Se colectaron 265 aislamientos de levaduras provenientes de hemocultivos, que se identificaron utilizando pruebas morfológicas, fisiológicas y bioquímicas y la determinación de la concentración inhibitoria mínima se realizó en base al estándar del NCCLS. La distribución de especies fue: Candida albicans (40,75%), Candida parapsilosis (28,67%), Candida tropicalis (15,84%), Candida famata (3,77%), Cryptococcus neoformans (3,77%), Candida glabrata (2,64%) y otras (4,53%). La mayoría de los aislamientos fueron sensibles a anfotericina B, fluconazol e itraconazol. La mortalidad asociada a las fungemias por levaduras estudiadas (n=265) fue del 30%, siendo más baja a lo descrito (33-54%) y fue menor en los pacientes que recibieron tratamiento antifúngico (26,3%), que en los no tratados (47%).

The incidence of candidemia has increased approximately 500% in high-complexity hospitals. A change in the spectrum of Candida infections due to species other than Candida albicans has also been detected. Between April 1999 and April 2000 a multicenter study was performed in order to determine the species distribution associated to candidemias in Argentina and the susceptibility profile of the isolates to the current antifungal drugs. Thirty six institutions have participated. All the 265 yeast strains isolated from blood cultures were identified by morphological, physiological, and biochemical tests. The antifungal susceptibility testing of isolates was performed based on the reference NCCLS procedure. The distribution of species was: Candida albicans (40.75%), Candida parapsilosis (28.67%), Candida tropicalis (15.84%), Candida famata (3.77%), Cryptococcus neoformans (3.77%), Candida glabrata (2.64%), and others (4.53%). Most of the isolates were susceptible to amphotericin B, fluconazole and itraconazole. Mortality associated to the fungemia by yeasts episodes (n=265) was 30%, lower than results previously determined (33-54%). The mortality percentage in patients who received antifungal therapy versus patients without treatment was 26.3% and 47%, respectively.

[First isolation of Histoplasma capsulatum from the urban bat Eumops bonariensis]. / Primer aislamiento de Histoplasma capsulatum de murciélago urbano Eumops bonariensis.

We report the first isolation of Histoplasma capsulatum var. capsulatum from a male bat Eumops bonariensis captured in Buenos Aires city in 2003. The pathogen was recovered from spleen and liver specimens, and was identified by its phenotypic characteristics. PCR with primers 1283, (GTG)5, (GACA)4 and M13 was used to compare both bat isolates with 17 human isolates, 12 from patients residing in Buenos Aires city, and 5 from other countries of the Americas. The profiles obtained with the four primers showed that both bat isolates were identical to each other and closer to Buenos Aires patients than to the other isolates (similarity percentage: 91-100% and 55-97%, respectively). The high genetic relationship between bat isolates and those from patients living in Buenos Aires suggests a common source of infection. This is the first record of E. bonariensis infected with H. capsulatum in the world, and the first isolation of the fungus in the Argentinean Chiroptera population. In the same way as these wild mammals act as reservoir and spread the fungus in the natural environment, infection in urban bats could well be associated with the increase in histoplasmosis clinical cases among immunosuppressed hosts in Buenos Aires city.

Primer aislamiento de Histoplasma capsulatum de murciélago urbano Eumops bonariensis / First isolation of Histoplasma capsulatum from the urban bat Eumops bonariensis

Se comunica el primer aislamiento de Histoplasma capsulatum var. capsulatum de un murciélago macho de la especie Eumops bonariensis, capturado en la ciudad de Buenos Aires en 2003. Los aislamientos fueron recuperados de bazo e hígado e identificados fenotípicamente. Se los comparó por PCR, con 17 aislamientos clínicos, 12 de pacientes residentes en la ciudad de Buenos Aires y cinco de otros países de América, usando los iniciadores 1283, (GTG)5, (GACA)4 y M13. Con los cuatro iniciadores, los perfiles de los aislamientos de murciélago resultaron idénticos entre sí y más relacionados a los de pacientes de Buenos Aires que a los de otros países (porcentaje de similitud: 91-100% y 55-87%, respectivamente). La alta relación genética entre los aislamientos obtenidos del murciélago y de los humanos residentes en Buenos Aires sugiere una fuente común de infección. Este es el primer registro de E. bonariensis infectado con H. capsulatum en el mundo, y el primer aislamiento del hongo en la población de quirópteros de la Argentina. Así como estos mamíferos actúan como reservorio y dispersan el hongo en la naturaleza, la infección en murciélagos urbanos podría asociarse al elevado número de casos de histoplasmosis entre pacientes inmunodeprimidos en la ciudad de Buenos Aires.

We report the first isolation of Histoplasma capsulatum var. capsulatum from a male bat Eumops bonariensis captured in Buenos Aires city in 2003. The pathogen was recovered from spleen and liver specimens, and was identified by its phenotypic characteristics. PCR with primers 1283, (GTG)5, (GACA)4 and M13 was used to compare both bat isolates with 17 human isolates, 12 from patients residing in Buenos Aires city, and 5 from other countries of the Americas. The profiles obtained with the four primers showed that both bat isolates were identical to each other and closer to Buenos Aires patients than to the other isolates (similarity percentage: 91-100% and 55-97%, respectively). The high genetic relationship between bat isolates and those from patients living in Buenos Aires suggests a common source of infection. This is the first record of E. bonariensis infected with H. capsulatum in the world, and the first isolation of the fungus in the Argentinean Chiroptera population. In the same way as these wild mammals act as reservoir and spread the fungus in the natural environment, infection in urban bats could well be associated with the increase in histoplasmosis clinical cases among immunosuppressed hosts in Buenos Aires city.

[Multicenter study of fungemia due to yeasts in Argentina]. / Estudio multicéntrico de fungemias por levaduras en la República Argentina.

The incidence of candidemia has increased approximately 500% in high-complexity hospitals. A change in the spectrum of Candida infections due to species other than Candida albicans has also been detected. Between April 1999 and April 2000 a multicenter study was performed in order to determine the species distribution associated to candidemias in Argentina and the susceptibility profile of the isolates to the current antifungal drugs. Thirty six institutions have participated. All the 265 yeast strains isolated from blood cultures were identified by morphological, physiological, and biochemical tests. The antifungal susceptibility testing of isolates was performed based on the reference NCCLS procedure. The distribution of species was: Candida albicans (40.75%), Candida parapsilosis (28.67%), Candida tropicalis (15.84%), Candida famata (3.77%), Cryptococcus neoformans (3.77%), Candida glabrata (2.64%), and others (4.53%). Most of the isolates were susceptible to amphotericin B, fluconazole and itraconazole. Mortality associated to the fungemia by yeasts episodes (n=265) was 30%, lower than results previously determined (33-54%). The mortality percentage in patients who received antifungal therapy versus patients without treatment was 26.3% and 47%, respectively.

Primer aislamiento de Histoplasma capsulatum de murciélago urbano Eumops bonariensis. / [First isolation of Histoplasma capsulatum from the urban bat Eumops bonariensis]

We report the first isolation of Histoplasma capsulatum var. capsulatum from a male bat Eumops bonariensis captured in Buenos Aires city in 2003. The pathogen was recovered from spleen and liver specimens, and was identified by its phenotypic characteristics. PCR with primers 1283, (GTG)5, (GACA)4 and M13 was used to compare both bat isolates with 17 human isolates, 12 from patients residing in Buenos Aires city, and 5 from other countries of the Americas. The profiles obtained with the four primers showed that both bat isolates were identical to each other and closer to Buenos Aires patients than to the other isolates (similarity percentage: 91-100

and 55-97

, respectively). The high genetic relationship between bat isolates and those from patients living in Buenos Aires suggests a common source of infection. This is the first record of E. bonariensis infected with H. capsulatum in the world, and the first isolation of the fungus in the Argentinean Chiroptera population. In the same way as these wild mammals act as reservoir and spread the fungus in the natural environment, infection in urban bats could well be associated with the increase in histoplasmosis clinical cases among immunosuppressed hosts in Buenos Aires city.

Estudio multicéntrico de fungemias por levaduras en la República Argentina. / [Multicenter study of fungemia due to yeasts in Argentina]

The incidence of candidemia has increased approximately 500

in high-complexity hospitals. A change in the spectrum of Candida infections due to species other than Candida albicans has also been detected. Between April 1999 and April 2000 a multicenter study was performed in order to determine the species distribution associated to candidemias in Argentina and the susceptibility profile of the isolates to the current antifungal drugs. Thirty six institutions have participated. All the 265 yeast strains isolated from blood cultures were identified by morphological, physiological, and biochemical tests. The antifungal susceptibility testing of isolates was performed based on the reference NCCLS procedure. The distribution of species was: Candida albicans (40.75

), Candida parapsilosis (28.67

), Candida tropicalis (15.84

), Candida famata (3.77

), Cryptococcus neoformans (3.77

), Candida glabrata (2.64

), and others (4.53

). Most of the isolates were susceptible to amphotericin B, fluconazole and itraconazole. Mortality associated to the fungemia by yeasts episodes (n=265) was 30

, lower than results previously determined (33-54

). The mortality percentage in patients who received antifungal therapy versus patients without treatment was 26.3

and 47

, respectively.