IN VITRO EVALUATION OF ANTHELMINTIC PROPERTIES OF THE METHANOLIC AND HYDRO ALCOHOLIC EXTRACTS OF ANNONA MURICATA ON HAEMONCHUS CONTORTUS.
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Date
2023-06
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SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA
Abstract
Haemonchus contortus is the most pathogenic nematode in sheep and has developed
resistance to all commercial anthelmintics throughout the world. In this context, the use
of medicinal plants represented as an alternative to chemical anthelmintics. The present
study was conducted to evaluate the in vitro anthelmintic effect of crude hydroalcoholic
(HE) and methanol extracts (ME) of Annona muricata fruit peel, seed and bark against
eggs, infective larvae, and adults of H. contortus by egg hatch assay (EHA), the larval
paralysis assay (LPA) and worm motility inhibition assay (WMIA) at different
concentrations (0.3 to 81 mg/mL) using albendazole as a reference drug. All extracts
exhibited significant (P<0.001) dose and or time dependant anthelmintic effects by
inhibiting egg hatching and causing larval paralysis, and mortality of worms compared
to negative control, and the effect was comparable with albendazole. At higher (81
mg/mL) concentration, A. muricata extracts showed 96 to 100% efficacy in EHA and
100% efficacy in LPA. In WMIA, worms were completely immobilized in extracts
within 1 h post-exposure. Overall performance of the hydroalcoholic extracts of the
plant on egg, larva, and adult H. contortus was better than the methanol extract of the
same plant except for the seed. At lower (9 and 3 mg/mL) concentrations extracts were
effective on egg and larval stage of H. contortus, but did not inhibit the motility of
worms. The respective effective dose 50% (ED50) with 95% confidence intervals were
estimated. The hydroalcoholic extracts of A. muricata fruit peel (ED50=1.28 mg/mL)
and seed (ED50=1.29 mg/mL) significantly inhibited egg hatching, whereas fruit peel
extracts significantly inhibited larval motility (ED50=2.71 mg/mL) and adult worm
motility (ED50=12.63 mg/mL), indicating their anthelmintic activity. Quantitative and
qualitative phytochemical screening of extracts revealed the presence of alkaloids,
flavonoids, tannins, amino acids, terpenoids and saponins, which could be accountable
for the anthelmintic activity observed. Alkaloids were exclusively present in fruit peel
HE. The results confirm the traditional use of A. muricata as a natural anthelmintic,
however, further in vivo studies are warranted to validate these effects