Distribution and growth pattern of Pseudomonas aeruginosa bacteria in the corneal stroma during corneal infection

Session Details

Session Title: Presented Poster Session: Infection & Imaging

Venue: Poster Village: Pod 2

First Author: : A.Elsahn UK

Co Author(s): :    M. Romero   M. Camara   H. Dua                 

Abstract Details

Purpose:

To study the behaviour of Pseudomonas aeruginosa bacteria inside the corneal stroma after corneal infection in human microbial keratitis

Setting:

Basic science laboratory study

Methods:

Human whole donor corneas obtained from the eye bank were used in this project. Bacterial suspensions of mCherry expressing P. aeruginosa PAO-1L with a fluorescence-based biosensors of c-di-GMP were prepared to a concentration of 107 cfu/ml. Fifty microlitres of bacterial suspension was injected into the middle of the corneal stroma from the endothelial using a 25G hypodermic needle. The corneas were incubated in medium for 72h and examined live under a Laser Confocal Fluorescence Microscopy (CFLM)

Results:

CFLM live imaging, mCherry-expressing PAO-1L bacteria were observed to grow locally at the site of injection. The levels of c-di-GMP initially increased steadily, indicating the increase of bacterial association. Towards the end of the 72h period, bacterial growth rate and the levels of c-di-GMP increased rapidly, and bacterial spread was observed in the corneal stroma

Conclusions:

We have established an ex-vivo models of bacterial keratitis and demonstrated that P. aeruginosa can grow rapidly within the corneal stroma after an initial phase of slow growth. This can implications on clinical decision making with regards to initiation and timing of antibiotic therapy

Financial Disclosure:

None