TY - JOUR
T1 - Microscopic characterization of biofilm in mixed keratitis in a novel murine model
AU - Ponce-Angulo, Diana Gabriela
AU - Bautista-Hernández, Luis Antonio
AU - Calvillo-Medina, Rosa Paulina
AU - Castro-Tecorral, Franco Ivan
AU - Aparicio-Ozores, Gerardo
AU - López-Villegas, Edgar Oliver
AU - Ribas-Aparicio, Rosa María
AU - Bautista-de Lucio, Victor Manuel
N1 - Publisher Copyright:
© 2019
PY - 2020/3
Y1 - 2020/3
N2 - Purpose: To report the characterization and analysis of the biofilm formation in mixed keratitis induced by the coinfection of Staphylococcus aureus and Fusarium falciforme in a novel murine model. Methods: Clinical ocular microbial isolates and female BALB/c mice were used to develop the murine model. Immunosuppression was achieved with cyclophosphamide and methylprednisolone. A corneoscleral lesion was performed with a micro-pocket technique. Mice received an inoculum with a concentration of 1 × 105 conidia of F. falciforme and S. aureus with 1 × 105 UFC/ml. Mice were sacrificed at 72 h after induction of infection, the right eye was enucleated and preserved in 10% formaldehyde to perform the PAS staining. In addition, cuts were obtained for the labeling with the fluorophores propidium iodide and Calcofluor White, and other eye cuts were processed to transmission microscopy. Results: F. falciforme and S. aureus were able to developed mono and mixed biofilm in vitro. Keratitis of F. falciforme, S. aureus and mixed, were established at immunosuppressed mice. Clinical symptoms were observed at murine cornea. Histological analysis by special stains identified bacterial, fungal and mixed biofilm structures at epithelial and stromal level. Extracellular matrix was observed surrounded clusters of bacterial, fungi and mixed by fluorescence and transmission electronic microscopy. Conclusion: This study provides direct evidence of the establishment and formation of mixed biofilm in vitro, as well as in vivo on the corneal surface of mice in an experimentally induced S. aureus and F. falciforme mixed keratitis infection.
AB - Purpose: To report the characterization and analysis of the biofilm formation in mixed keratitis induced by the coinfection of Staphylococcus aureus and Fusarium falciforme in a novel murine model. Methods: Clinical ocular microbial isolates and female BALB/c mice were used to develop the murine model. Immunosuppression was achieved with cyclophosphamide and methylprednisolone. A corneoscleral lesion was performed with a micro-pocket technique. Mice received an inoculum with a concentration of 1 × 105 conidia of F. falciforme and S. aureus with 1 × 105 UFC/ml. Mice were sacrificed at 72 h after induction of infection, the right eye was enucleated and preserved in 10% formaldehyde to perform the PAS staining. In addition, cuts were obtained for the labeling with the fluorophores propidium iodide and Calcofluor White, and other eye cuts were processed to transmission microscopy. Results: F. falciforme and S. aureus were able to developed mono and mixed biofilm in vitro. Keratitis of F. falciforme, S. aureus and mixed, were established at immunosuppressed mice. Clinical symptoms were observed at murine cornea. Histological analysis by special stains identified bacterial, fungal and mixed biofilm structures at epithelial and stromal level. Extracellular matrix was observed surrounded clusters of bacterial, fungi and mixed by fluorescence and transmission electronic microscopy. Conclusion: This study provides direct evidence of the establishment and formation of mixed biofilm in vitro, as well as in vivo on the corneal surface of mice in an experimentally induced S. aureus and F. falciforme mixed keratitis infection.
KW - Biofilm
KW - Fungus-bacteria interactions
KW - Mixed keratitis
UR - http://www.scopus.com/inward/record.url?scp=85078077920&partnerID=8YFLogxK
U2 - 10.1016/j.micpath.2019.103953
DO - 10.1016/j.micpath.2019.103953
M3 - Artículo
C2 - 31904447
AN - SCOPUS:85078077920
SN - 0882-4010
VL - 140
JO - Microbial Pathogenesis
JF - Microbial Pathogenesis
M1 - 103953
ER -