An elderly female presented to the ophthalmology Department, TUTH, Kathmandu, Nepal with red painful eyes. She was treated for all viral, fungal, bacterial keratitis without any improvement. As the patient had severe pain with ring ulcerative lesion of cornea, ophthalmologists suspected it to be Acanthamoeba keratitis. Although, Acanthamoeba had never been successfully cultured previously in Nepal, ophthalmologists requested me and my colleague to try it out. So, we collected the corneal scraping of the patient, taken by the ophthalmologist. We prepared a wet mount for microscopy from the sample. On microscopy, we could see few cystic and other irregular structures. However, we were not able to know what it was. Then, we prepared a media for Acanthamoeba culture called non-nutrient media (media with salts and agar) which was lawn cultured with Escherichia coli.
We cultured the corneal scraping in a C-pattern in the media and incubated it at 37 degree Celsius for 24 hours. Next day, when the plates were observed, we could see some growth around the inoculated area but colonies were not visible. So, I focused the plate directly on microscope in 4x and 10x where colonies became visible. The colonies were resembling that of Acanthamoeba. We prepared wet mount from the grown culture which revealed cysts of Acanthamoeba. For further confirmation Gram stain, Giemsa stain and Trichome stain was done. After the microscopic examination of the stained smears, we could confirm that the culture growth was that of Acanthamoeba species.
Since, Polyhexamethylen biguanide (PHMB) which is the drug of choice was not available in Nepal. Patient was not improving with other medical treatment and finally keratoplasty was done by which she improved
Thanks to Meenu mam for her guidance and Dr. Shusila for her support.