First report of Acanthamoebae spp. isolation from a volcanic mud spring in the Philippines.

Acanthamoebae spp. is considered highly adaptive. The present study aims to establish the occurrence of free-living amoebae, particularly Acanthamoebae, to exist in extreme environments such as volcanic mud springs. Fifty surface water samples were collected from mud springs (34 samples), and flat rocks (16 samples) were collected, processed, and cultured. After 14 days of incubation, 32 (64%) plates showed positive amoebic growth. Nineteen (55.8%) of these plates came from the mud spring collection site, while 13 (81.2%) samples are from flat rock sources. DNAs from positive samples were made to react to polymerase chain reaction (PCR) using primer sets JDP1 5'GGCCCAGATCGTTTACCGTGAA-3' and JDP2 5'TCTCACAAGCTGCTAGGGAGTCA-3' for cells that resemble Acanthamoebae. Sequencing and basic local alignment search tool (BLAST) revealed a 99% similarity of isolates to Acanthamoebae spp. Identification of Acanthamoebae spp that can survive in higher temperatures is important public health information. The existence of such isolates in the environment has dire health implications, which suggests revisitation of water treatment protocols. Detection of such organisms in environmental sources used for recreational purposes provides information to local and international tourists who frequent them. This will result in the mitigation of potential future infection.