A preliminary assessment of the feasibility of evaluating promising antifilarials in vitro against adult Onchocerca volvulus

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Abstract
The suitability of motility indices and tetrazolium-based colorimetric assays for the determination of the viability of adult Onchocerca volvulus after in vitro exposure to potential macrofilaricides has been examined. Experimentation showed that both techniques could be applied to adult O. volvulus, although the variability between individual worms necessitated the use of large experimental groups. The potential of using cut anterior tips of female O. volvulus for screening was also investigated. These were shown to give reasonably consistent motility indices, and drug effects were discernible even after 72 h in vitro culture. Application of these viability criteria to studies on the short-term in vitro survival of intact male and female O. volvulus incubated in Eagles MEM plus serum, under 5% CO2 in air, showed this medium to be suboptimal with a greater than 50 % loss of worm viability within 144 h of nodulectomy. Males isolated by the collagenase technique were shown to be significantly less viable than dissected males, by both motility indices and tetrazolium reduction. The results highlight the need to use either dissected males, or in the case of females, the need to minimize exposure to collagenase solution. A possible mechanism for selecting a more uniformly viable female worm population is discussed. Examination of the in vitro effects of CGP 20376 using these viability criteria/assay systems showed some delayed suppression of worm motility, but after 120 h in vitro CGP 20376 was not macrofilaricidal against male or female O. volvulus. Male worms were also implanted subcutaneously into gerbils. Treatment of these animals with CGP 20376, with autopsy 2 weeks later, resulted in a small macrofilaricidal effect (28% worm death). Some reduction in the motility of living worms recovered from CGP 20376-treated gerbils was also evident. Good survival of control worms recovered 2 weeks post-implantation suggests that male O. volvulus might survive longer subcutaneously. The prospects for undertaking meaningful drug evaluations against O. volvulus in vitro (and in vivo in rodents) is therefore promising, but because of parasite availability will probably be limited to those compounds which have shown potential in other filarial test systems.