Recent evidence suggests that eye-seeking flies are important trachoma vectors. We conducted a series of investigations to identify which species of synanthropic flies are potential vector(s) of this blinding disease in The Gambia. Several species of fly were caught in fish-baited attractant traps placed in villages throughout the year (1997/98) but only 2 species, Musca sorbens and M. domestica, were caught from the eyes of children. M. sorbens comprised < 10% of the total number of flies caught with attractant traps but was responsible for > 90% of fly-eye contacts, the remainder were made by M. domestica. All fly species were more numerous in the wet season than the dry season. Eyes of young children are considered to be the main reservoir of Chlamydia trachomatis, the causative agent of trachoma. Collections of eye-seeking flies from children showed frequent fly-eye contacts (median [interquartile range], 3 [1.5-7] every 15 min). Children with potentially infective ocular or nasal discharge had twice as many fly-eye contacts than children with no discharge (P < 0.001). There was no difference in exposure to fly-eye contacts if a child sat inside or outside a house (P = 0.273). Female flies were more commonly caught from eyes than male (P < 0.001). The presence of Chlamydia DNA was demonstrated by PCR on 2 of 395 flies caught from the eyes of children with a current active trachoma infection. Both positive flies were M. sorbens, one male and the other female. Further elucidation of M. sorbens behavioural ecology and the development of sustainable strategies to control these flies should be a priority. It is likely that M. sorbens is the principal insect vector of trachoma in The Gambia.