Trials were carried out to study the humoral immune response of blackflies to filariae following infection using the intrathoracic injection technique. An induced 66 kDa protein was abundant in the haemolymph of the European species Simulium ornatum following infection with bovine Onchocerca lienalis. This protein was apparently at higher concentrations in the haemolymph of sham-inoculated flies, i.e. flies that received sterile medium without the parasites. A molecule of the same size was also observed in the haemolymph of infected S. damnosum s.l. following infection with human O. volvulus or bovine O. ochengi. However, the level of this protein was lower in blackflies injected with microfilariae of bovine O. dukei. Unlike O. volvulus and O. ochengi this species is not transmitted by S. damnosum s.l. under natural conditions. No such reaction was observed if the African blackflies had received a sham inoculation. Feeding experiments with wild-caught nulliparous S. damnosum sl. on Onchocerca-infected cattle supported the results of the injection trials. The 66 kDa protein could only be found in the haemolymph of specimens infected via a blood meal. This 66 kDa molecule was identified as phenoloxidase. It appeared in the haemolymph due to the activation of the prophenoloxidase system following the filarial infection and we hypothesize that it may be sequestered by the parasites, as part of a non-self recognition system.

Onchocerca volvulus exists in at least two strains in West Africa, while its black-fly vectors consist of sibling species, dwelling in the savanna and forest/transition zones. In transition and degraded forest zones both parasite strains and different sibling species of the vector can be sympatric. The strain of parasite in infected humans and in vector black-flies was determined in two bioclimes along the Bandama river of Côte d'Ivoire. The upper Bandama is located in the savanna bioclime while the Middle Bandama is located in a degraded forest zone. At both sites, savanna-dwelling sibling species of the Simulium damnosum sensu lato species complex predominated. The severe-strain of O. volvulus was the predominant strain at both sites. However, severe-strain parasites represented a significantly larger proportion of those found in the vector population than in the human population in the degraded forest of the Middle Bandama. These data suggest that in degraded forest areas recently invaded by savanna-dwelling species of S. damnosum s.l. transmission of the severe-strain of the parasite might be more efficient than transmission of the mild-strain.

Injection trials with compatible and non-compatible Onchocerca species into S. damnosum s.l., the vector of human and bovine onchocerciasis, demonstrated that the rapid killing of microfilariae within the blackfly's haemocoel is species specific. In the presence of the peptide RGDS as a blocking agent for integrin-like receptors of haemocytes, the survival of O. ochengi microfilariae in its natural intermediate host was significantly increased. This increased survival 24 h p.i. correlated with a significant decrease of apoptosis levels in the microfilariae following a 2 h exposure to the haemolymph in vivo. These findings suggest that haemocytes are directly involved in the killing of Onchocerca microfilariae in the blackfly.