Acacia nilotica fruits with high tannin content are used in the northern parts of Cameroon as anti-filarial remedies by traditional healers. In this study, the hydro-alcoholic fruit extract (crude extract (CE)) and, one of the main constituents in its most active fractions, (+)-catechin-3-O-gallate (CG), as well as four related proanthocyanidins, (−)-epicatechin-3-O-gallate (ECG), (+)-gallocatechin (GC), (−)-epigallocatechin (EGC) and (−)-epigallocatechin-3-O-gallate (EGCG), were assessed for their potential in vitro anthelmintic properties against the free-living model organism Caenorhabditis elegans and against the cattle filarial parasite Onchocerca ochengi. Worms were incubated in the presence of different concentrations of fruit extract, fractions and pure compounds. The effects on mortality were monitored after 48 h. The plant extract and all of the pure tested compounds were active against O. ochengi (LC50 ranging from 1.2 to 11.5 µg/mL on males) and C. elegans (LC50 ranging from 33.8 to 350 µg/mL on wild type). While high LC50 were required for the effects of the compounds on C. elegans, very low LC50 were required against O. ochengi. Importantly, tests for acute oral toxicity (lowest dose: 10 mg/kg) in Wistar rats demonstrated that crude extract and pure compounds were non-toxic and safe to use. Additionally, the results of cytotoxicity tests with the Caco-2 cell line (CC50 ranging from 47.1 to 93.2 µg/mL) confirmed the absence of significant toxicity of the crude extract and pure compounds. These results are in good accordance with the use of A. nilotica against nematode infections by traditional healers, herdsmen and pastoralists in Cameroon.
14 August 2017 in BMC Complementary and Alternative Medicine
Onchocerciasis 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. Plant 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. Ethanolic 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. Ethanolic 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.
The front line molecules from filarial worms and other nematodes or helminthes are their Excretory-Secretory (ES) products. Their interaction with the host cells, proteins and immune system accounts for the skin and eye pathology or hyposensitivity observed in human onchocerciasis. ES products and adult worms’ crude extracts from Onchocerca ochengi, a filarial nematode that infects the African zebu cattle, were utilized in the present study as a model for studying Onchocerca volvulus that causes river blindness in man. The ES products were generated from adult male and female worms in vitro and analyzed with poly acrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA) using sera from Onchocerca-infected cattle and humans. The cattle sera were collected from a herd that had been exposed for six years to natural transmission of Onchocerca spp. The expressed reactivity was evaluated and differences analyzed statistically using Kruskal-Wallis rank and Chi-square tests. The gel electrophoretic analyses of 156 ES products from O. ochengi female and male worms and of two somatic extracts from three females and 25 males revealed differences in the protein pattern showing pronounced bands at 15, 30–50 and 75 kDa for male ES proteins and 15, 25 and 40–75 kDa for somatic extracts, respectively and less than 100 kDa for female worms. Proteins in the ES products and somatic extracts from female and male Onchocerca ochengi worms were recognized by IgG in sera from both Onchocerca-exposed cattle and humans. Bovine serum antibodies reacted more strongly with proteins in the somatic extracts than with those in the ES products. Interestingly, the reaction was higher with male ES products than with ES products from female worms, suggesting that the males which migrate from one nodule to another are more exposed to the host immune system than the females which remain encapsulated in intradermal nodules. This study demonstrates that O. ochengi ES products and, in particular, extracts from male filariae may represent a good source of immunogenic proteins and potential vaccine candidates.
10 January 2017 in British Journal of Pharmaceutical Research
One of the strategies for developing novel pharmaceutical drugs is to use natural sources such as plants for therapeutic treatment. Plant extracts are a cocktail of compounds which act synergically and can improve treatment effectiveness, reduce therapeutic duration and resistance. The ethanolic extracts of leaves and seeds of Cucurbita pepo oviferavar ovifera from Sudano-Guinean and Sudano-Sahelian zones of Cameroon were evaluated on the cattle parasite nematode Onchocerca ochengi. Worms were incubated with different concentrations of the plant extracts in RPMI-1640 supplemented with streptomycin and gentamicin. Mortality at 37°C was monitored after 24, 48 and 72 h. Ivermectin was used as positive control and DMSO as negative. Plant extracts from the two ecological zones showed anthelminthic activities on O. ochengi after 72 h with LC50 varying from 20 to 1090 µg /mL. The highest antifilarial activity in Sudano-Guinean zone was obtained with leave extract of C. pepo ovifera (LC50 of 20 µg/mL), while highest antifilarial activity in Sudano-Sahelian zone was obtained with seed extracts of the plant with LC50 value of 17 µg/mL after 72 h. These results show that anthelmintic activity depends on the part of the plant and the ecological zones. Additionally, the plant is not toxic. These results on the ethanolic extracts of leaves and seeds of C. pepo ovifera var ovifera confirmed the use of this plant in traditional medicine in Cameroon to treat disease due to nematodes. The plants could be used as alternative anthelmintic to fight against Human and Bovine onchocerciasis.
10 January 2017 in Asian Journal of Medicine and Health
Aim: The objective was to evaluate the anthelmintic activity of the ethanolic stem bark of A. occidentale. Study Design: Ethanolic extract of stem barks of Anacardium occidentale was tested in vitro against the cattle filarial parasite Onchocerca ochengi and the wild type, the drug resistant strains of the free living nematode Caenorhabditis elegans. Place and Duration of Study: The work took place at the Laboratory of Applied Zoology of the Faculty of the University of Ngaoundere between May 2016 and March 2017. Methodology: Adults of O. ochengi were incubated in RPMI-1640 medium supplemented with penicillin/streptomycin and plant extract or drugs. C. elegans was incubated at 20°C with plant extract or drugs in M9-medium. Worm mortality was determined biochemically by MTT/formazan colorimetry after 24, 48 and 72 h. Ivermectin and levamisole were used as positive control and DMSO as negative control. Acute toxicity on mice (Mus musculus) and phytochemical compounds were also determined. Results: Ethonolic stem bark extract (SBE) of Anacardium occidentale was active against worms. O. ochengi was the most affected by the extract with the LC50 values of 88 ± 0.16 µg/mL, 3.18 ±0.17 µg/mL and 2.76 ± 0.10 µg/mL respectively after 24, 48 and 72h of incubation. The quantitative phyto-constituents screening has revealed the presence of tannins, phenolic acids, saponins and flavonoids. Tannins (107.16 mg GAE/ 100 g) were the most quantified compound followed by phenolic acid (26.23 mg/g GAE), saponins (9.33 mg/g) and flavonoids (0.16 mg of rutin/100 g). The acute toxicity study has demonstrated that at short term (14 days), oral intake of ethanolic stem bark extract was not toxic up to a dose of 1600 mg/kg by weight. Conclusion: This study justifies the use of this plant by traditional health practitioners in managing the disease, and also suggests a source of compounds against Onchocerca volvulus.