info:eu-repo/semantics/article
Separation of Paracoccidioides brasiliensis conidia through Percoll gradients
Registro en:
1369-3786
10.1080/13693780410001657126
1460-2709
Autor
Restrepo Moreno, Ángela
Cano Restrepo, Luz Elena
Jiménez Alzate, María del Pilar
García Moreno, Luis Fernando
Institución
Resumen
ABSTRACT: The conidia of Paracoccidioides brasiliensis are the structures most likely to serve
as the infectious propagules of this fungus. This study describes our attempts to
purify conidia by eliminating mycelial fragments. Purification was attempted using
discontinuous 95% and 60% Percoll gradients with densities of 1.167 and 1.107,
respectively, prepared either in 0.15 mol/L PBS or 0.25 mol/L sucrose. The best
results were observed with the 95% and 90% gradients in sucrose; with the former,
conidial purity ranged from 70.6 to 100%, with a mean of 82.3% and a coefficient
of variation (VC) of 11.7. With 90% gradients, purity was achieved between 70.4
and 92.5%. The mean in this case was 80.6% and the VC was 9.2%. The use of two
consecutive 95% Percoll gradients in sucrose was tested. The recovery efficiency per
plate, which averaged 2.5 /106 conidia per plate with one gradient, increased to
5.19/1.3 /106 conidia with two gradients. The use of Percoll did not affect the
viability of the conidia, which was always]/90%. This method allows the
preparation of a conidial sample almost free from contamination with mycelial
fragments, thus facilitating quantitative determination of cause and effect in in-vivo
interactions between P. brasiliensis and its hosts. COL0013709 COL0008639 COL0013746