Comparison of axenic and monoxenic media for isolation of Acanthamoeba

Acanthamoeba is a genus of ubiquitous, free-living amebae that can be difficult to isolate by standard microbiologic techniques. We retrospectively reviewed the laboratory records of patients with ocular acanthamoebic infection for the period from January 1973 to June 1996 and found that Acanthamoeba isolates were recovered from 73, 71, and 70% of clinical specimens inoculated onto buffered charcoal-yeast extract agar (BCYE), nonnutrient agar with live or dead Escherichia coli, and tryptic soy agar (TSA) with horse or sheep blood, respectively. We then prospectively compared the recovery of a corneal isolate of Acanthamoeba on commercial media from Remel and BBL (TSA with 5% sheep blood, TSA with 5% horse blood, TSA with 5% rabbit blood, V agar, chocolate agar, BCYE, and selective BCYE with polymyxin B, anisomycin, and vancomycin) and on axenic and monoxenic media prepared with live or dead bacteria (Enterobacter aerogenes, E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, and Stenotrophomonas maltophilia). Good recovery of trophozoites was obtained on BCYE, TSA with rabbit blood, TSA with horse blood, and Remel TSA with sheep blood. BBL TSA with horse blood or rabbit blood provided good recovery of cysts. All species of live or dead bacteria yielded good recovery of trophozoites; however, only nonnutrient agar with live P. aeruginosa, live E. aerogenes, or live S. maltophilia gave good recovery of cysts. TSA with either rabbit blood or horse blood, BCYE, and nonnutrient agar prepared with live P. aeruginosa, E. aerogenes, or S. maltophilia offer optimal recovery of Acanthamoeba.