Original
Actividad proteinasa y fosfolipasa como factores de virulencia en especies de Candida aisladas de sangreProteinase and phospholipase activity as virulence factors in Candida species isolated from blood

https://doi.org/10.1016/S1130-1406(08)70050-0Get rights and content

Resumen

El número de fungemias nosocomiales debidas a especies de Candida ha aumentado en las últimas décadas. Para producir una infección, los patógenos oportunistas tienen que evadir el sistema inmunológico, sobrevivir y dividirse en los tejidos del huésped y diseminarse a otros órganos. La secreción de proteinasa y fosfolipasa ha sido descrita como factor potencial de virulencia de algunas especies de Candida responsables de candidemias en pacientes con catéteres ingresados en las unidades de cuidados intensivos. En este trabajo se ha tratado de demostrar que la secreción de los enzimas proteinasa y fosfolipasa son factores de virulencia de especies de Candida aisladas de hemocultivos de pacientes admitidos en las unidades de cuidados intensivos, unidades de diálisis y unidades de oncología. Ciento catorce aislamientos de Candida fueron obtenidos de muestras de sangre; se identificaron las siguientes especies: Candida albicans (37), Candida glabrata (7), Candida guilliermondii (5), Candida kefyr (3), Candida krusei (45), Candida parapsilosis (5) y Candida tropicales (12). La detección de proteinasa fue realizada utilizando el método de Staib, y la de fosfolipasa utilizando el método de Samaranayake et al. Se determinó la zona de precipitación (valor de Pz). El porcentaje de aislamientos que produjo cantidades perceptibles de proteinasa fue 74,56%; los aislamientos productores de fosfolipasa fueron el 44,73%. En el presente estudio se pudo observar que la producción de proteinasa y fosfolipasa puede ser un factor importante de virulencia en las fungemias causadas por Candida independientemente de la especie que haya sido aislada.

Summary

The number of nosocomial blood stream infections due to Candida species has increased over the past few decades. In order to establish an infection, opportunistic pathogens have to evade the immune system, survive, divide in the host environment, and spread to other tissues. Proteinase and phospholipase secretion has been implicated as potential virulence factors for some Candida species responsible for catheter related candidemia in intensive care unit (ICU) patients with indwelling devices. We therefore have aimed at demonstrating the secretion of proteinase and phospholipase enzymes as virulent factors by Candida species isolated from blood samples collected from ICUs, dialysis units and oncology units. One hundred and fourteen isolates of Candida species were obtained from the blood samples and the isolates include 37 Candida albicans, 7 Candida glabrata, 5 Candida guilliermondii, 3 Candida kefyr, 45 Candida krusei, 5 Candida parapsilosis, and 12 Candida tropicalis. Proteinase assay was performed by using the Staib method. Phospholipase assay was performed by using the method of Samaranayake et al. Precipitation zone (Pz value) was determined. The percentage of isolates which produced detectable amounts of proteinase is 74.56% and 44.73% of isolates produced detectable amounts of phospholipase. We believe that production of both phospholipase and proteinase enzimes could be an important virulence factor for several Candida species.

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    Citation Excerpt :

    Afterwards, all the plates were incubated at 37 °C for five days and fixed with 20% trichloroacetic acid and stained with 1.25% amidoblack (Merck, Germany). The Proteinase production (Prz) was determined by measuring the ratio of the diameter around the well to the diameter of the proteolytic non-stained zone [28,29]. The C. albicans ATCC 10231 and C. albicans PFCC 9362 were cultured on Sabouraud dextrose agar (SDA) (Merck, Germany) at 37 °C for 24 h, To have a high logarithmic growth number of organisms, cell cultures were incubated overnight at 25 °C (shaking at 110 oscillations per minute) in yeast extract peptone dextrose broth.

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