IN VITRO EVALUATION OF ANTHELMINTIC PROPERTIES OF THE HYDRO-ALCOHOLIC EXTRACTS OF SELECTED MEDICINAL PLANTS ON Haemonchus contortus
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Date
2018-12
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SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA
Abstract
In recent years, there has been growing interest in alternative therapies
and the therapeutic use of natural products especially of medicinal plants for control of
parasites. In the present study in vitro assays such as egg hatch assay (EHA), larval
paralysis assay (LPA) and adult worm motility inhibition assay (WMIA) were
conducted to determine the anthelmintic efficiency of hydro-alcoholic extracts of
Anacardium occidentale shell, Illicium verum fruit and Artocarpus heterophyllus seed
on eggs, infective larvae (L3) and adult worms of Haemonchu contortus, in comparison
to albendazole. Among three extracts, A. occidentale shell showed significant (P<0.01)
inhibitory effect on egg hatching and larval motility followed by A. heterophyllus seed
and I. verum fruit extracts. Extracts of A. occidentale shell induced 50% inhibition at
lower concentration (0.0255 mg/mL) compared to I. verum fruit extract (0.0612
mg/mL) and A. heterophyllus seed (0.0372 mg/mL) extracts. The LD50 value of
reference drug albendazole (positive control) was 0.237 µg/mL. Extracts of A.
occidentale shell required maximum of 0.5 mg/mL, whereas extracts of I. verum fruit
and A. heterophyllus seed required maximum concentration of 4 and 2 mg/mL
respectively, to induce 100 per cent egg hatch inhibition. Similarly A. occidentale shell
showed maximum activity on motility of L3 larvae (LD50 = 0.196 mg/mL) with 100 per
cent paralysis while A. heterophyllus seed (LD50 = 0.666 mg/mL) and I. verum fruit
(LD50 = 1.418 mg/mL) exhibited 84.67±1.76 and 72.66±1.76 per cent paralysis
respectively, at higher tested concentration of 6 mg/mL. In WMIA, three extracts
induced significant (P<0.001) mortality of adult worms; however the activity of A.
occidentale shell was higher (100%) than I. verum fruit (36.6±3.3%) and A.
heterophyllus seed (70.00±5.7%) at a concentration of 6 mg/mL within 1 h post
exposure. Anacardium occidentale extract revealed better LD50 (1.0365 mg/mL) values
in comparison with I. verum fruit (LD50 = 3.848 mg/mL) and A. heterophyllus seed
(LD50 = 2.398 mg/mL) in the WMIA. Three plant extracts exhibited significant
(P<0.001) dose dependant anthelmintic responses by inhibiting egg hatching and
causing paralysis of larvae and mortality of worms. In vitro effect of these extracts on
lactate dehydrogenase activity of H. contortus was also studied. All extracts
significantly (P<0.01) inhibited the activity of LDH catalysing the oxidation of lactate.
Maximum level of inhibition of LDH activity was noticed in A. occidentale shell extract
treated worms. Thin Layer Chromatography analysis and qualitative phytochemical
screening of three extracts revealed presence of alkaloids, flavonoids, tannins, saponins,
carbohydrates and proteins which might be responsible for the anthelmintic effects
noticed. Overall, this in vitro study suggests that these three plants have promising
anthelmintic effects.
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