In- vitro evaluation of antithelminthic activity of albendazole and three midicinal plant extracts on Capillaria sp and its characterization with RAPD-PCR

Capillaria species are nematodes of serious fish diseases which may be transferred to human. The present study aimed to evaluat the anthelminthic activity of different concentrations of albendazole and three medicinal plants (Allium sativum, Lupinus termis, and Cucrbita pepo)on Capillaria sp. Infested fresh water catfishes, Bagrus docmac and B.bayad.collected from Menoufia and Kalyobia Governorates, Egypt. Five different media; RPMI 1640(Formula 1), RPMI 1640 (Formula 11), RPMI 1640 (Formula 111), Hank`s saline and Natural calf serum in optimum conditions were used for in vitro maintaining of Capillaria sp. Natural calf serum was the best medium for maintenance of Capillaria sp. as observed by high activity and low mortality rate in optimum conditions for 24 hours (pH; 7 & 20°C). Albendazole at a dose of 5.5x106 ppm showed high mortality rate (86.6%) after 24 hours exposure time. Water extracts of the medicinal plants (A.sativum, l.termis, and C.pepo) have higher effect on the mortality rate of adult Capillaria sp. in vitro than albendazole. A.sativum water extract was high effective than L.termis and C. pepo extracts where lower concentration (18x103 ppm) gave maximum mortality rate (100%) 24 hours post exposure. DNA of Capillaria sp. was isolated and the RAPD-PCR was applied. using 6 arbitrary primers: [G-11b (TGCCCGTGG), G-15b (ACTGGGACTC), G-18b (GGCTCATCTC), M-1(AGGTCACTGA), M-are(ATCTGGCAAC),G-7b (GAACCTGCGG) ].The obtained result indicated that RAPD-PCR assay can be used to differentiate Capillaria sp. from its geneticallay close species of the superfamily Trichuroidea in which genera Trichinella, Trichuris and Capillaria are included. RAPD-PCR has proved useful to detect genomic instability and DNA variation induced by treatments of sublethal concentrations (LC50) of albendazole and Allium sativum water extract.