Cyclospora spp. in Drills, Bioko Island, Equatorial Guinea

To the Editor: More than a decade has passed since major outbreaks of Cyclospora cayetanensis infection in the United States and Canada drew attention to this newly emerging infection (1,2). Awareness of these infections was highlighted again by large outbreaks in the summer of 2013 (3). However, many questions remain unanswered regarding this organism, including aspects of its life cycle, geographic distribution, and range of related species. In 1999, three new distinct Cy-clospora species noted for their close similarity with C. cayetanensis from humans were isolated from monkeys in Ethiopia (4). A survey of primates in Kenya increased awareness of the extended distribution of these 3 species in eastern Africa and provided confirmation of their marked host specificity, even where the ranges of host species overlapped (5). Most recently , C. colobi–like organisms were identified in snub-nosed golden colo-bus monkeys in northwestern China (6). We report the characterization of Cyclospora spp. recovered from drills (Mandrillus leucophaeus poensis) on Bioko Island, Equatorial Guinea. During January–February 2011 and 2012, fecal samples from free-ranging animals were collected and placed in 10% formalin (2011) or potassium dichromate (2012). Because samples were collected opportunisti-cally from unidentified animals of un-determined age and sex, whether any samples were collected from the same animals in either year was not known. Fecal samples were concentrated by using the formyl ethyl acetate method , and sediment was examined by using fluorescent microscopy to detect oocysts (4). Three (9%) of 26 samples from 2011 and 8 (31%) of 25 samples from 2012 were positive for Cyclospora oocysts that were spherical, measured 8–10 µm in diameter, and showed au-tofluorescence. The oocysts collected in potassium dichromate had sporulat-ed by the time of examination, which facilitated and confirmed identification as Cyclospora spp. Representative samples from 4 animals in the second collection were submitted for molecular analysis. The entire 18S rRNA gene (1,796 bp) was obtained from 2 DNA fragments amplified by PCR from DNA extracted from 3 fecal specimens by using procedures and primers for genetic analysis of coccidian parasites (4,7,8). Six distinct full-length 18S rRNA sequences were obtained and compared with sequences in GenBank. Although our sequences showed high similarity with 18S rRNA genes for all Cyclospora species, the sequences were most similar to the C. papionis 18S rRNA gene (GenBank accession no. AF111187), even though 3 T → C transitions at nucleotides positions 680, 1054, and 1694 were observed. Analyses of these …