Abstract
BackgroundOnchocerciasis is one of the tropical neglected diseases (NTDs) caused by the nematode Onchocerca volvulus. Control strategies currently in use rely on mass administration of ivermectin, which has marked activity against microfilariae. Furthermore, the development of resistance to ivermectin was observed. Since vaccine and safe macrofilaricidal treatment against onchocerciasis are still lacking, there is an urgent need to discover novel drugs. This study was undertaken to investigate the anthelmintic activity of Lophira lanceolata on the cattle parasite Onchocerca ochengi and the anthelmintic drug resistant strains of the free living nematode Caenorhabditis elegans and to determine the phytochemical profiles of the extracts and fractions of the plants.MethodsPlant was extracted in ethanol or methanol-methylene chloride. O. ochengi, C. elegans wild-type and C. elegans drug resistant strains were cultured in RPMI-1640 and NGM-agar respectively. Drugs diluted in dimethylsulphoxide/RPMI or M9-Buffer were added in assays and monitored at 48 h and 72 h. Worm viability was determined by using the MTT/formazan colorimetric method. Polyphenol, tannin and flavonoid contents were determined by dosage of gallic acid and rutin. Acute oral toxicity was evaluated using Swiss albino mice.ResultsEthanolic and methanolic-methylene chloride extracts killed O. ochengi with LC50 values of 9.76, 8.05, 6.39 μg/mL and 9.45, 7.95, 6.39 μg/mL respectively for leaves, trunk bark and root bark after 72 h. The lowest concentrations required to kill 50% of the wild-type of C. elegans were 1200 and 1890 μg/mL with ethanolic crude extract, 1000 and 2030 μg/mL with MeOH-CH2Cl2 for root bark and trunk bark of L. lanceolata, respectively after 72 h. Leave extracts of L. lanceolata are lethal to albendazole and ivermectin resistant strains of C. elegans after 72 h. Methanol/methylene chloride extracted more metabolites. Additionally, extracts could be considered relatively safe.ConclusionEthanolic and methanolic-methylene chloride crude extracts and fractions of L. lanceolata showed in vitro anthelmintic activity. The extracts and fractions contained polyphenols, tannins, flavonoids and saponins. The mechanism of action of this plant could be different from that of albendazole and ivermectin. These results confirm the use of L. lanceolata by traditional healers for the treatment of worm infections.
Highlights
Onchocerciasis is one of the tropical neglected diseases (NTDs) caused by the nematode Onchocerca volvulus
Anthelmintic activity of ethanolic and methanolic/ methylene chloride extracts of L. lanceolata on O. ochengi The anthelmintic activities of leaves, trunk bark and root bark of L. lanceolata on O. ochengi adult and on C. elegans wild type (WT) were evaluated in terms of mortality after 48 h and 72 h of incubation
Positive controls were strongly active against O. ochengi with lowest concentrations required to inhibit 50% mortality (LC50) of 2.23 ± 1.96 μg/mL for ivermectin, 3.62 ± 1.88 μg/mL, for levamisole and 4.34 ± 0.71 μg/mL for albendazole after 72 h incubation (Table 1)
Summary
Onchocerciasis is one of the tropical neglected diseases (NTDs) caused by the nematode Onchocerca volvulus. This study was undertaken to investigate the anthelmintic activity of Lophira lanceolata on the cattle parasite Onchocerca ochengi and the anthelmintic drug resistant strains of the free living nematode Caenorhabditis elegans and to determine the phytochemical profiles of the extracts and fractions of the plants. The control started with vector control involving spraying of insecticides and larvicides [7] followed by mass treatment using various combinations of drugs including ivermectin which is the recommended molecule against onchocerciasis [8] This drug reduces significantly transmission of the disease, its filaricidal effect is limited only to the juvenile form of the parasite [9]. The present study investigates the in vitro antifilarial activity of both crude extracts and chromatographic fractions of extracts of L. lanceolata leaves, trunk bark and root bark against O. ochengi adult forms, C. elegans wild type as well as drug resistant strains. We investigated the acute toxicity and the phytochemical profiles of the extracts and fractions of the plants
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