Fungizone amphotericin b

African green monkey kidney BSC-40 and VERO cells (American type cell culture—ATCC) were maintained in 5% CO₂ atmosphere at 37 °C in Eagle’s Minimum Essential Medium (MEM) (Gibco BRL, Invitrogen, Carlsbad, CA, USA) supplemented with 5% fetal bovine serum (FBS) (Cultilab, Brazil), 25 µg/mL fungizone (Amphotericin B) (Cristália, São Paulo, Brazil), 500 U/mL penicillin (Cristália, São Paulo, São Paulo, Brazil) and 50 µg/mL gentamicin (Schering-Plough, São Paulo, Brazil). VERO cells were used for viral isolation from clinical specimens and growth. The BSC-40 cells were used for viral clones purification, plaque phenotype, comet phenotype and growth curve assays. Only typical poxviruses cytopathic effects were visualized after inoculation of clinical samples in both cell systems. The VACV Western Reserve (VACV-WR) was gently provided by Dr C. Jungwirth (Universität Würzburg, Würzburg, Germany) and was used as a virulent control in mice assays.

African green monkey kidney BSC-40 (ATCC-CRL-2761) and VERO (ATCC-CCL-81) cells were maintained in a 5% CO₂ atmosphere at 37 °C, in Eagle’s Minimum Essential Medium (MEM) (Gibco BRL, Invitrogen, Carlsbad, CA, USA), supplemented with 5% fetal bovine serum (FBS) (Cultilab, Brazil), 25 µg/mL fungizone (Amphotericin B) (Cristália, São Paulo, Brazil), 500 U/mL penicillin and 50 µg/mL gentamicin (Schering-Plough, São Paulo, Brazil). VERO cells were used for viral replication and the BSC-40 cells were used for viral plaque phenotypes and titration. The VACV used in the study, PV1 and PV2, were isolated from clinical specimens of horses during an equine vaccinia outbreak [15 (link),19 (link)].