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Originally published In Press as doi:10.1167/iovs.08-2358 on October 3, 2008
 (Investigative Ophthalmology and Visual Science. 2009;50:729-736.)
© 2009 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.08-2358
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Pathogenic Phenotype and Genotype of Pseudomonas aeruginosa Isolates from Spontaneous Canine Ocular Infections

Eric C. Ledbetter,1 James J. Mun,2 David Kowbel,2 and Suzanne M. J. Fleiszig2

1From the College of Veterinary Medicine, Department of Clinical Sciences, Cornell University, Ithaca, New York; and the 2School of Optometry, Division of Infectious Diseases, University of California at Berkeley, Berkeley, California.

PURPOSE. This study was designed to determine whether the ability to adversely affect corneal epithelial cell health is a factor common to Pseudomonas aeruginosa keratitis strains and to assess the prevalence of each pathogenic phenotype and genotype in a canine model of naturally-acquired P. aeruginosa ocular infection.

METHODS. P. aeruginosa ocular isolates were collected by sampling 100 dogs without disease (six isolates collected) and by sampling dogs with conjunctivitis (two isolates), endophthalmitis (one isolate), active keratitis (12 isolates), and resolved P. aeruginosa keratitis (four isolates). Phenotype was determined in vitro by quantifying corneal epithelial cell invasion by gentamicin survival assays, and cytotoxic activity by Trypan blue exclusion assays. Genotyping was performed for genes encoding the type III secreted effectors.

RESULTS. The ratio of invasive to cytotoxic strains with 95% confidence intervals (CI) was 0.83 (CI, 0.42–0.99) for conjunctival microflora isolates, 0.80 (CI, 0.54–0.94) for ocular infection isolates, and 1.0 (CI, 0.45–1.0) for strains isolated post-resolution of keratitis. Among ocular infection isolates, invasive and cytotoxic strains were significantly (P ≤ 0.02) associated with older and younger dogs, respectively. Visible adverse effects on epithelial cells were significantly (P ≤ 0.03) more frequent for keratitis strains (6/12) than other strains (1/13), but only three of these keratitis strains and the single non-keratitis strain possessed ExoU.

CONCLUSIONS. Invasive strains predominated in the dogs of this study. Only keratitis strains had visible adverse effects on epithelial cells without overt cytotoxicity, suggesting virulence strategies affecting live corneal epithelial cell health are selected for among keratitis strains.






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