Efficacy of transport media use versus direct inoculation of blood agar plates in the microbiologic evaluation of experimental Streptococcus pneumoniae keratitis.

Purpose: To compare the microbiologic yield of cultures obtained by direct inoculation of blood agar plates (BAP) from corneal ulcer swabbings versus indirect inoculation via transport media in a rabbit model of Streptococcus pneumoniae bacterial keratitis.
Methods: The corneas of 12 rabbits were inoculated with S. pneumoniae. Keratitis was confirmed 18 hours later. Sampling was performed at four 2.5-hour intervals. At each interval, corneal swabs were directly applied to BAP and placed into transport medium: thioglycollate and Amies medium without charcoal. Swabbings were then subcultured onto BAP at two time points: 2 and 24 hours after collection in transport medium. Plates were evaluated 48 hours later. Organism recovery rates were measured in terms of the number of positive culture plates observed and the bacterial colony counts on each plate.
Results: The rate of positive cultures overall was 69%. The recovery rates were similar for direct inoculation, inoculation via Amies held for 2 hours, and inoculation via Amies held for 24 hours. Direct inoculation yielded fewer colonies than indirect inoculation via Amies held for 24 hours (p = 0.008). Direct inoculation yielded a higher rate of positive cultures than did thioglycollate held for 2 hours (p = 0.004) or 24 hours (p < 0.001). The rate of nonpneumococcal contaminants ranged from 6% of BAP subcultured from thioglycollate held for 24 hours to 28% of directly inoculated BAP.
Conclusions: Amies medium without charcoal may be used as a transport medium for up to 24 hours in the recovery of S. pneumoniae from corneal ulcers in this rabbit model. Thioglycollate appears to be less effective as a transport medium. Results of this study may justify studies of other transport media and/or other corneal pathogens. Altogether, such studies may provide justification for human clinical trials.