July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Microscopic characterization of mouse model mixed keratitis
Author Affiliations & Notes
  • Diana Gabriela Ponce-Angulo
    Microbiology and Ocular Proteomics , Intituto de Oftamología Conde de Valenciana, Mexico City, Mexico
    Producción y Control de Biológicos, Escuela Nacional de Ciencias Biológicas-IPN, Mexico City, Mexico
  • Luis Antonio Bautista-Hernández
    Microbiology and Ocular Proteomics , Intituto de Oftamología Conde de Valenciana, Mexico City, Mexico
  • Rosa Paulina Calvillo-Medina
    Microbiology and Ocular Proteomics , Intituto de Oftamología Conde de Valenciana, Mexico City, Mexico
  • Gerardo Aparicio-Ozores
    Bacteriología Médica, Escuela Nacional de Ciencias Biológicas-IPN, Mexico City, Mexico
  • Rosa María Ribas-Jaimes
    Producción y Control de Biológicos, Escuela Nacional de Ciencias Biológicas-IPN, Mexico City, Mexico
  • Victor Manuel Bautista
    Microbiology and Ocular Proteomics , Intituto de Oftamología Conde de Valenciana, Mexico City, Mexico
  • Footnotes
    Commercial Relationships   Diana Gabriela Ponce-Angulo, None; Luis Antonio Bautista-Hernández, None; Rosa Paulina Calvillo-Medina, None; Gerardo Aparicio-Ozores, None; Rosa María Ribas-Jaimes, None; Victor Bautista, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 506. doi:
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      Diana Gabriela Ponce-Angulo, Luis Antonio Bautista-Hernández, Rosa Paulina Calvillo-Medina, Gerardo Aparicio-Ozores, Rosa María Ribas-Jaimes, Victor Manuel Bautista; Microscopic characterization of mouse model mixed keratitis. Invest. Ophthalmol. Vis. Sci. 2018;59(9):506.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Corneal infections have been described originated by bacteria, fungi, virus, and co-infection, which can involve more than two infectious agents. In the world, more than two millions two cases have been reported. There are few reports of polimicrobial infections in ophthalmology, however, cases are increasing every year. Several inveestigations have been developed an effective animal model in order to study and understand co-infectious keratitis physiology. Based on these information the aim of this study was performed a mixed keratitis murine model of Staphylococcus aureus and Fusarium falciforme and microscopic characterization.

Methods : We performed an immunosuppressants ascending concentration curve (methylprednisolone-cyclophosphamide), using ocular clinical strains and BALB / c female mice at 6-8 weeks old. We performed an ocular injury by the pocket technique. The rigth eyes were inoculated with Fusarium falciforme (1x105 conidia) and S. aureus (1X105 UFC). The mice were sacrificed at 72 h after induction of infection, the right eyes were enucleated and preserved with formaldehyde. Histological preparations were developed and stained with H-E and PAS stains. In another hand, stained preparations with calcofluor white and propidium iodide were made and analyzed by fluorescence microccopy and finally transmission microscopy was performed to observed the microorganisms prescence and morphology.

Results : It is important to mention that this is the first experimental work where used a combination of two immunosuppressants cyclophosphamide-methylprednisolone obtaining saticfactory results with cyclophosphamide 150 mg/kg and methylprednisolones 80 mg/kg of mouse weight. We found that the microbial distribution was in the cornea at stroma level and epithelium. In some cases the F. falciforme hyphae were located at lens level, the previous observations were revealed by histopathological and fluorescence analysis and finally the transmission electron microscopy suggested biofilm formation by S. aureus and F. falciforme in mono and mixed infection.

Conclusions : The microbial distribution was determinated by fluorescence and histopalogic analysis observing that the S. aureus had a dsitribution at epithelial level while F. falciforme was found at stromal, retina and lens level. We found that both microorganisms infected the mice cornea and induced a mixed keratitis and biofilm formation.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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