Three sympatrically occurring members of a cichlid fish species complex from Lake Malawi have previously been reported to show assortative mating in the field. Offspring from three experimental tanks each housing both sexes of all three members of this complex were screened at one to four microsatellite loci to determine paternity. Reproductive isolation was found to persist without exception under laboratory conditions, providing strong evidence for behavioural barriers to mating. Direct mate choice is suggested as the process preventing interbreeding. The results are interpreted with reference to the rapid and explosive evolution of the cichlid group.