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Sandra Huérfano

Publications and source records attributed to Sandra Huérfano.

3 recordsLinked to original sources

[Isolation of Cryptococcus gattii serotype B from detritus of Eucalyptus trees in Colombia].

INTRODUCTION: Cryptococcus gattii serotype B has been associated with several species of trees in tropical and subtropical regions. Serotype B has been found to be the second most frequently isolated from patients in Colombia, but it has not been isolated from the environment. Therefore, a study was designed to sample plant material from a forest in the province of Cundinamarca, with the aim of establishing the presence and distribution of the C. neoformans complex, specially serotype B. In addition, isolates were evaluated for several phenotypic characteristics associated with virulence. METHODOLOGY: During February and March, 2003, 4 separate sample collections were made. One hundred sixty-seven samples were collected from detritus of eucalyptus and 28 samples from other species of trees. All were processed using the conventional procedure and plated on selective media. The following parameters were measured: cellular and capsular sizes in the filtrates, growth at 37 degrees C, virulence in a mouse animal model system, and mating type as detected by PCR. RESULTS: Cryptococcus gattii serotype B was isolated from 46 eucalyptus samples (27.5%). The cellular size in the filtrates ranged from 3.10 to 4.15 microm and the capsular size from 0.30 and 0.46 microm. The 46 isolates grew at 37 degrees C, but none of them caused the death in the animals during 70-day observation period. All isolates were of mating type a. CONCLUSIONS: This constitutes the first report of the isolation of serotype B, mating type a, from eucalyptus detritus in Colombia.

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[Phenotype characterization of environmental Cryptococcus neoformans isolates].

Cryptococcosis is caused by the three varieties of C. neoformans with physiological and virulence differences, some of which have been studied to determine biological aspects of this microorganism. The phenotypical aspects of environmental isolates from varieties grubii and gattii were evaluated to establish differences associated with their life cycle and virulence. To this end, 28 and 31 strains of C. neoformans serotypes A (var. grubii) and C (var. gattii) were studied. The microscopic and macroscopic morphology on Sabouraud agar and soils, growth rate at 37 degrees C, production of 22 extracellular enzymes, haploid fructification, mating type, killer toxin sensitivity patterns and virulence in BALB/c mice were evaluated. No differences were observed between the two varieties regarding microscopic and macroscopic morphology or growth at 37 degrees C (p > 0.05). However, a decrease in the cellular and capsular sizes of yeast in soil, as compared to Sabouraud, was observed (p < 0.05). Additionally, higher enzimatic activity of proteases, phospholipases, phenoloxidase and beta-glucosidase was observed in var. grubii isolates as compared to var. gattii (p < 0.05). In both varieties, structures related with haploid fruitification were observed and all isolates were mating type alpha. Killer toxin sensitivity patterns of the isolates of var. grubii were I and II; in contrast, in var. gattii, seven different patterns were found: I, V, IX-XIII. In the animal model we found that 12 of 22 (54.5%) isolates of var. grubii caused the death of the mice during the observation period, while none of the 14 var. gattii isolates caused it. The decrease in capsular and cellular sizes of the yeast in soil and the frequency of mating type alpha with structures related to haploid fructification suggest an important mechanism of production of infectious particles in nature. Additionally, greater enzimatic activity of var. grubii can be associated with the virulence in the animal model.

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[Experimental inoculation of Terminalia catappa seedlings with an environmental isolate of Cryptococcus neoformans var. gattii serotype C ].

In 1997, our laboratory reported for the first time the isolation of Cryptococcus neoformans var. gattii serotype C associated with almond tree (Terminalia catappa) detritus. This finding led to a more detailed follow up of the association between the plant and the yeast. Preliminary data have shown that survival of the yeast in almond trees seedlings goes beyond 100 days. The aim of the present study was to establish if under the conditions previously studied, C. neoformans var. gattii would remain viable for longer periods. A total of 83 almond tree seedings, 20-40 cm high, were inoculated with C. neoformans var. gattii serotype C (INS-755). Assays were carried out inoculating the stem or the soil where the seedlings were planted. Observations were undertaken for a period of up to 12 months. As processing techniques we employed the endophytic fungi procedure (stems), maceration (roots, leaves) and standard suspension method (soils). Additionally, microscopic visualization of the yeast in plant tissues was done with trypan blue plus lactophenol. C. neoformans var. gattii was recovered from the inoculated plants for a period of up to 12 months post-inoculation; additionally, the fungus had the capacity to migrate from the stem to the soil and viceversa, without causing macroscopic or microscopic alterations in the plant tissues. This finding suggests that there appears to be an association between the host plant and C. neoformans var. gattii in the environment.

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