Blood culture as a parameter of treatment effectiveness in experimental histoplasmosis of the hamster Hemocultivos como parametro de la eficacia del tratamiento de la histoplasmosis experimental en hamster

The aim of this study was to determine the value of blood culture as a parameter of treatment effectiveness in experimental histoplasmosis. A total of thirty five hamsters, weighing approximately 120g, were inoculated intracardiacly with 0.1 ml of a suspension containing 4 x 10(7) cells/ml of the yeast phase of H. capsulatum. Treatments were started one week after the infection and lasted for 3 weeks. The azoles, (itraconazole, saperconazole and fluconazole) were administered once a day by gavage, at a dose of 8 mg/kg; Amphotericin B was given intraperitonealy every other day at a dose of 6mg/kg. Blood samples (1 ml) were obtained by heart punction from the 4th day after infection and were seeded in Sabouraud honey-agar and BHI-agar. The hamsters that survived were killed one week after treatment completion and the following criteria were considered for treatment evaluation: 1) rate of spontaneous death, at the end of the experience; 2) microscopic examination of Giemsa smears from liver and spleen and 3) determination of CFU in spleen cultures. Amphotericin B was the most effective drug, with negative blood cultures at day 20, negative spleen cultures in all cases and all the animals survived until the end of the study. Fluconazole was the less effective drug, blood cultures were positive during the whole experience, spleen cultures showed a similar average of CFU when compared with the control animals and 42.8% of these animals died. Saperconazole and itraconazole showed a similar activity, with survival of all hamsters and negative blood cultures at 23 and 26 days respectively. Blood culture seems to be valuable parameter for treatments' evaluation in experimental histoplasmosis of the hamster.<br>El propósito de esta investigación fue determinar el valor de los hemocultivos para juzgar la eficacia de los tratamientos en la histoplasmosis experimental. Treinta y cinco hamsters fueron inoculados por via intracardíaca con 0.1 ml de una suspensión de la fase levaduriforme del H. capsulatum equivalente a 4 x 10(7) células/ml. Los tratamientos comenzaron 1 semana después de la infección y se mantuvieron durante 3 semanas. Los compuestos azólicos, itraconazol, saperconazol y fluconazol se administraron por gastroclisis, una vez al día, a razón de 8mg/kg de peso; la anfotericina B fue aplicada por via intraperitoneal día por medio, en dosis de 6mg/kg. Se obtuvieron muestras de sangre para hemocultivo (1 ml) a partir del 4º día post-infección y se sembraron placas de agar-miel de Sabouraud y agar infusión de cerebro y corazón. Los animales que sobrevivieron fueron autopsiados y se tomaron en consideración los siguientes parámetros: 1) tasa de muertes espontáneas al finalizar la experiencia; 2) resultados de los exámenes microscópicos de extendidos de hígado y bazo teñidos com Giemsa y; 3) UFC en cultivos de bazo. La anfotericina B fue la droga más eficaz, los hemocultivos de los animales tratados fueron negativos a partir del 20º dia de tratamiento, se negativizaron los cultivos de bazo y la totalidad de los hamsters sobrevivió. El menos eficaz fue el fluconazol que no modificó los resultados de los hemocultivos y los cultivos esplénicos y se registró el 42.8% de muertes espontáneas durante el estudio. Saperconazol e itraconazol exhibieron una eficacia similar, no se observaron muertes en los animales tratados y los hemocultivos se negativizaron a los 23 y 26 dias de tratamiento respectivamente. Los hemocultivos resultaron ser útiles como criterio para evaluar la eficacia terapéutica de las drogas antifúngicas en este modelo experimental de histoplasmosis

Hemocultivos como parametro de la eficacia del tratamiento de la histoplasmosis experimental en hamster

The aim of this study was to determine the value of blood culture as a parameter of treatment effectiveness in experimental histoplasmosis. A total of thirty five hamsters, weighing approximately 120g, were inoculated intracardiacly with 0.1 ml of a suspension containing 4 x 10(7) cells/ml of the yeast phase of H. capsulatum. Treatments were started one week after the infection and lasted for 3 weeks. The azoles, (itraconazole, saperconazole and fluconazole) were administered once a day by gavage, at a dose of 8 mg/kg; Amphotericin B was given intraperitonealy every other day at a dose of 6mg/kg. Blood samples (1 ml) were obtained by heart punction from the 4th day after infection and were seeded in Sabouraud honey-agar and BHI-agar. The hamsters that survived were killed one week after treatment completion and the following criteria were considered for treatment evaluation: 1) rate of spontaneous death, at the end of the experience; 2) microscopic examination of Giemsa smears from liver and spleen and 3) determination of CFU in spleen cultures. Amphotericin B was the most effective drug, with negative blood cultures at day 20, negative spleen cultures in all cases and all the animals survived until the end of the study. Fluconazole was the less effective drug, blood cultures were positive during the whole experience, spleen cultures showed a similar average of CFU when compared with the control animals and 42.8% of these animals died. Saperconazole and itraconazole showed a similar activity, with survival of all hamsters and negative blood cultures at 23 and 26 days respectively. Blood culture seems to be valuable parameter for treatments' evaluation in experimental histoplasmosis of the hamster.El propósito de esta investigación fue determinar el valor de los hemocultivos para juzgar la eficacia de los tratamientos en la histoplasmosis experimental. Treinta y cinco hamsters fueron inoculados por via intracardíaca con 0.1 ml de una suspensión de la fase levaduriforme del H. capsulatum equivalente a 4 x 10(7) células/ml. Los tratamientos comenzaron 1 semana después de la infección y se mantuvieron durante 3 semanas. Los compuestos azólicos, itraconazol, saperconazol y fluconazol se administraron por gastroclisis, una vez al día, a razón de 8mg/kg de peso; la anfotericina B fue aplicada por via intraperitoneal día por medio, en dosis de 6mg/kg. Se obtuvieron muestras de sangre para hemocultivo (1 ml) a partir del 4º día post-infección y se sembraron placas de agar-miel de Sabouraud y agar infusión de cerebro y corazón. Los animales que sobrevivieron fueron autopsiados y se tomaron en consideración los siguientes parámetros: 1) tasa de muertes espontáneas al finalizar la experiencia; 2) resultados de los exámenes microscópicos de extendidos de hígado y bazo teñidos com Giemsa y; 3) UFC en cultivos de bazo. La anfotericina B fue la droga más eficaz, los hemocultivos de los animales tratados fueron negativos a partir del 20º dia de tratamiento, se negativizaron los cultivos de bazo y la totalidad de los hamsters sobrevivió. El menos eficaz fue el fluconazol que no modificó los resultados de los hemocultivos y los cultivos esplénicos y se registró el 42.8% de muertes espontáneas durante el estudio. Saperconazol e itraconazol exhibieron una eficacia similar, no se observaron muertes en los animales tratados y los hemocultivos se negativizaron a los 23 y 26 dias de tratamiento respectivamente. Los hemocultivos resultaron ser útiles como criterio para evaluar la eficacia terapéutica de las drogas antifúngicas en este modelo experimental de histoplasmosis

Comparación de métodos de extracción de ADN de sangre para detectar ADN fúngico por PCR Comparison of different methods of DNA extraction from blood to detect fungal DNA by PCR

La infección fúngica invasora (IFI) está asociada a un alto índice de mortalidad, que alcanza el 50% debido a la frecuente falla en el tratamiento antifúngico. Existen dificultades para realizar un diagnóstico micológico rápido y certero dada la baja sensibilidad de los métodos convencionales, especialmente en pacientes neutropénicos y con SIDA. Numerosos métodos para diagnosticar infecciones micóticas basados en el estudio del ADN fúngico están actualmente en desarrollo. Nosotros evaluamos la utilidad de dos procedimientos de extracción y purificación del ADN fúngico presente en sangre para su posterior detección por PCR. Ambos métodos resultaron igualmente eficientes para obtener ADNs de óptima calidad y para realizar la técnica de PCR con los iniciadores universales para hongos ITS 1 e ITS 4.Invasive fungal infections (IFI) are associated with high mortality by reaching levels of 50%, and also with a significant failure in antifungical treatments. This fact mostly obeys to difficulties in obtaining a fast and accurate mycologic diagnosis due to the low sensitivity of conventional methods, mainly in neutropenic and AIDS patients. Various methods based on fungal DNA study are currently being used for the diagnosis of mycotic infections. We herein evaluated two procedures of extraction and purification of fungal DNA in blood for their use in PCR detection. Both of them showed equal efficiency in obtaining high performance DNA with universal primers ITS 1and ITS 4 as target

Combination antifungal therapy: A strategy for the management of invasive fungal infections

The lack of an ideal antifungal drug or completely successful agent, suggests that combination therapy may be an appropriate option for the management of fungal infections. In this review, different antifungal combination therapy approaches will be discussed. In brief, the rationale for combination therapy is to maximize antifungal effects by attacking different fungal targets at the same time. However, the scientific basis for this approach requires evidence from prospective clinical trials of antifungal combinations. It is possible that real advantages will be seen for particular combinations only in particular mycoses and/or particular types of patient. Combination tends to reduce clinical failure when resistant strains could be recovered from patients, although drug interactions and cross-resistance may result. Synergy has been established between conventional antifungal agents and also between investigational molecules. Alternatively, combination of fluconazole and cyclosporine results fungicidal for fluconazole-susceptible strains of Candida albicans. An overall enhanced susceptibility is reached in intrinsically resistant Candida when combination of azole antifungal drugs or terbinafine with calcineurin inhibitors is used. Azoles, such as voriconazole, itraconazole, and fluconazole are being tested in combination with other antifungal agents such as amphotericin B, caspofungin, or terbinafine and. In addition, amphotericin B is actually administered in combined therapy with caspofungin. The traditional combinations of amphotericin B plus 5-fluorocytosine or amphotericin B plus rifampicin have been replaced by these newer combinations. Other combinations such as caspofungin plus amphotericin B or voriconazole have also been synergic against clinical isolates of Aspergillus and Fusarium. This has been confirmed in animal models in the case of caspofungin plus itraconazole in the treatment of invasive pulmonary aspergillosis and caspofungin plus liposomal amphotericin B in the management of invasive aspergillosis.Fil: Carrillo Muñoz, A. J.. Departamento de Micología; EspañaFil: Finquelievich, Jorge. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Tur Tur, Cristina. Departamento de Micología; EspañaFil: Eraso, Elena. Universidad del Pais Vasco; EspañaFil: Jaureguizar, Nerea. Universidad del Pais Vasco; EspañaFil: Quindós, Guillermo. Universidad del Pais Vasco; EspañaFil: Giusiano, Gustavo Emilio. Universidad Nacional del Nordeste. Instituto de Medicina Regional; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Molecular epidemiology of Aspergillus species and other moulds in respiratory samples from Argentinean patients with cystic fibrosis

In cystic fibrosis (CF) patients, fungal colonization of the respiratory tract is frequently found. Aspergillus fumigatus is the most frequently recorded and is associated with loss of pulmonary function and allergic disease (ABPA). The knowledge on prevalence rates of filamentous fungi in CF patients in Latin America is scarce. One hundred and seventy-six fungal isolates recovered from the upper respiratory tract of CF patients from Argentina were identified to species by morphology and DNA sequencing. In total, 90% of CF patients were colonized by Aspergillus sp., followed by Exophiala sp. (14%) and Scedosporium sp. (10%). Among Aspergillus, six species complexes (Fumigati, Flavi, Terrei, Nigri, Usti, and Nidulante) and different cryptospecies were found. Among Scedosporium, three species were observed (Scedosporium apiospermum, Scedosporium aurantiacum and Scedosporium boydii). All Exophiala isolates were identified as Exophiala dermatitidis. Rare filamentous fungi were also found. All cases of ABPA were associated to the presence of A. fumigatus. Mixed colonization with other mould or rare fungi was observed in half of them. To our knowledge, this is the first prospective study of mould species in CF using molecular methods in Latin America. This study shows that Aspergillus sp., E. dermatitidis and Scedosporium sp. have a high frequency in CF patients from Argentina, and by far, A. fumigatus was the most commonly cultured species. Continuous clinical surveillance is required to detect the emergence of new fungal pathogens and to detect resistant or difficult-to-treat species capable of chronic colonizing the airways and of hematogenous dissemination in case of lung transplantation.Fil: Brito Devoto, Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Hermida Alava, Katherine Stefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; ArgentinaFil: Pola, Santiago J.. Universidad de Buenos Aires; ArgentinaFil: Pereda, Rosana. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños Pedro Elizalde (ex Casa Cuna); ArgentinaFil: Rubeglio, Etelvina. Universidad de Buenos Aires; ArgentinaFil: Finquelievich, Jorge L.. Universidad de Buenos Aires; ArgentinaFil: Cuestas, María Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentin

Genetic diversity and population structure of teosinte

We examined data from the ARTEMIS DISK Antifungal Surveillance Program to describe geographic and temporal trends in the isolation of Candida parapsilosis from clinical specimens and the in vitro susceptibilities of 9,371 isolates to fluconazole and voriconazole. We also report the in vitro susceptibility of bloodstream infection (BSI) isolates of C. parapsilosis to the echinocandins, anidulafungin, caspofungin, and micafungin. C. parapsilosis represented 6.6% of the 141,383 isolates of Candida collected from 2001 to 2005 and was most common among isolates from North America (14.3%) and Latin America (9.9%). High levels of susceptibility to both fluconazole (90.8 to 95.8%) and voriconazole (95.3 to 98.1%) were observed in all geographic regions with the exception of the Africa and Middle East region (79.3 and 85.8% susceptible to fluconazole and voriconazole, respectively). C. parapsilosis was most often isolated from blood and skin and/or soft tissue specimens and from patients hospitalized in the medical, surgical, intensive care unit (ICU) and dermatology services. Notably, isolates from the surgical ICU were the least susceptible to fluconazole (86.3%). There was no evidence of increasing azole resistance over time among C. parapsilosis isolates tested from 2001 to 2005. Of BSI isolates tested against the three echinocandins, 92, 99, and 100% were inhibited by concentrations of ?2 ?g/ml of anidulafungin (621 isolates tested), caspofungin (1,447 isolates tested), and micafungin (539 isolates tested), respectively. C. parapsilosis is a ubiquitous pathogen that remains susceptible to the azoles and echinocandins; however, both the frequency of isolation and the resistance of C. parapsilosis to fluconazole and voriconazole may vary by geographic region and clinical service. Copyright " 2008, American Society for Microbiology. All Rights Reserved.",,,,,,"10.1128/JCM.02122-07",,,"http://hdl.handle.net/20.500.12104/41744","http://www.scopus.com/inward/record.url?eid=2-s2.0-40749148116&partnerID=40&md5=9835815df8990f312e2e431e71b96e85",,,,,,"3",,"Journal of Clinical Microbiology",,"84