In vitro and in vivo anthelmintic activity of Euphorbia helioscopia L.

doi:10.1016/j.vetpar.2012.04.023

Veterinary Parasitology

Volume 189, Issues 2–4, 26 October 2012, Pages 317-321

https://doi.org/10.1016/j.vetpar.2012.04.023 Get rights and content

Abstract

The anthelmintic potentials of the aqueous and methanol extracts of Euphorbia helioscopia were investigated. In folklore medicine, E. helioscopia (Euphorbiaceae) is used in the treatment of various gastrointestinal ailments and diseases. A worm motility inhibition (WMI) assay and egg hatch assay (EHA) were used for in vitro assessment, and a faecal egg count reduction (FECR) assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude methanolic extracts of E. helioscopia on live Haemonchus contortus worms as evident from their paralysis and/or death at 8 h after exposure, different concentrations (12.5 mg ml⁻¹, 25 mg ml⁻¹ and 50 mg ml⁻¹) of aqueous and methanolic extracts were used against H. contortus which exhibited dose-dependent anthelmintic effects on H. contortus. Aqueous and methanolic extracts of aerial parts (stem, leaves and flowers) of E. helioscopia were found to have very low percent inhibitory egg hatching as compared to levamisole. It is concluded that the entire plant of E. helioscopia possesses significant anthelmintic activity and could be a potential alternative for treating cases of helminth infections in ruminants.

Section snippets

Collection of plant materials

The present plant material, E. helioscopia (Local name Guir sochel) (Euphorbiaceae), is frequently seen throughout the Kashmir Valley in areas of high altitude – wastelands, agricultural lands and on open mountainous slopes. It is usually erect, slender stemmed, spreading up to 80 cm tall, though sometimes it can be seen lying down. The plant is an annual broad-leaved herb that has a hairy stem with many branches from the base to the top. Leaves are opposite, elliptical, oblong or

Faecal egg count reduction test

The in vivo anthelmintic activity (in terms of egg count percent reduction) of methanolic extracts of E. helioscopia in sheep naturally infected with mixed species of GI nematodes demonstrated significant anthelmintic activity of extracts tested. Both crude aqueous and crude methanol extracts of E. helioscopia exhibited a dose-dependent anthelmintic activity (Table 1). The maximum reduction (86.07%) in faecal egg counts was recorded for methanolic extract followed by aqueous extract of E.

Discussion

For the effective chemotherapy and strategic chemprophylaxis of Haemonchus infection, safe drugs are required with high activity against all stages of H. contortus. Hence, the development of a newer, safe, curative and economical drug has remained an active area of research. The present data clearly show that E. helioscopia is effective against H. contortus as is evident from FECR rate and larval mortality of nematodes. On comparing the methanolic and aqueous extracts, highest FECR rate

Conclusion

Based on the results of the present study, it can be concluded that E. helioscopia aerial parts (stem, leaves and flowers) tested in the form of crude aqueous and methanolic extracts showed significant in vitro and in vivo anthelmintic activity at concentrations and doses tested against ovine nematodes as determined by motility inhibition, reduction in faecal egg counts and egg hatch of H. contortus of sheep. These findings suggest that E. helioscopia could form an alternative to commercially

Acknowledgements

The authors thankfully acknowledge the financial assistance of the Department of Science and Technology, Govt. of India, New Delhi in conducting this study under the research project. Authors are thankful to Prof. Azra N. Kamili, Director Centre of Research for Development, University of Kashmir, Srinagar for providing necessary laboratory facilities during extraction.

References (35)

M.S. Akhtar et al.
Anthelmintic activity of medicinal plants with particular reference to their use in animals in Indo-Pakistan subcontinent
Small Rumin. Res.
(2000)
L.M. Assis et al.
Ovicidal and larvicidal activity in vitro of Spigelia anthelmia Linn. extracts on Haemonchus contortus
Vet. Parasitol.
(2003)
E.S. Bizimenyera et al.
In vitro activity of Peltophourum africanum Sond, (Fabaceae) extracts on the egg hatching and larval development of the parasitic nematode Trichostrongylus colubrifrmis
Vet. Parasitol.
(2006)
G.C. Coles et al.
World Association for the Advancement of Veterinary Parasitology (W.A.A.P). Methods for detection of anthelmintic resistance in nematodes of veterinary importance
Vet. Parasitol.
(1992)
G.C. Coles et al.
The detection of anthelmintic resistance in nematodes of veterinary importance
Rev. Vet. Parasitol.
(2006)
T. Eguale et al.
Haemonchus contortus: in vitro and in vivo anthelmintic activity of aqueous and hydro-alcoholic extracts of Hedera helix
Exp. Parasitol.
(2007)
P. Hordegen et al.
In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay
J. Ethnopharmacol.
(2006)
P. Hordegen et al.
The anthelmintic efficacy of five plant products against gastrointestinal trichostrongyloids in artificially infected lambs
Vet. Parasitol.
(2003)
A. Hussain et al.
An account of the botanical anthelmintics used in traditional veterinary practices in Sahiwal district of Punjab
Pakistan J. Ethnopharmacol.
(2008)
A. Jabbar et al.
Anthelmintic activity of Chenopodium album (L.) and Caesalpinia crista (L.) against trichostrongylid nematodes of sheep
J. Ethnopharmacol.
(2007)

J.K. Ketzis et al.

Chenopodium ambrosioides and its essential oil as treatments for Haemonchus contortus and mixed adult-nematode infections in goats

Small Rumin. Res.

(2002)

M.V. Maciel et al.

Ovicidal and larvicidal of Melia azedarach extracts on Haemonchus contortus

Vet. Parasitol.

(2006)

S.M.K. Rates

Plants as source of drugs

Toxic

(2001)

K.A. Tariq et al.

Anthelmintic activity of extracts of Artemisia absinthium against ovine nematodes

Vet. Parasitol.

(2009)

P.J. Waller

The development of anthelmintic resistance in ruminant livestock

Acta Trop.

(1994)

P.J. Waller

International approaches to the concept of integrated control of nematode parasites of livestock

Int. J. Parasitol.

(1999)

C.L. Marley et al.

The effect of birds foot trefoil (Lotus corniculatus) and chicory (Cichorium intybus) on parasite intensities and performance of lambs naturally infected with Helminth parasites

Vet. Parasitol.

(2003)

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Fourteen previously undescribed diterpenoids, including an unusual diterpenoid (1) with a 9,10-seco-jatrophane skeleton, ten jatrophane-type diterpenoids (2–11), two lathyrane-type diterpenoids (12, 13), and an abietane-type diterpenoid (14), together with thirty-six known ones (15–50), were isolated from the whole plants of Euphorbia helioscopia L. The structures of the new isolates were characterized by spectroscopic methods, single-crystal X-ray diffraction analysis, and computational prediction of ECD and chemical shifts. Thirty-nine abundant diterpenoids were evaluated for their enhancement of NK cell-mediated killing of NSCLC cells. As a result, compounds 24, 33, and 41 were found to significantly enhance the killing activity of NK cells towards H1299-luci cells and A549-luci cells at the concentration of 2.5 μM.
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The LC50 of water and methanolic extracts on egg hatching of Haemonchus contortus were 2.13 and 4.48 mg/mL compared with that of levamisole (1.88 mg/mL). In an faecal egg count reduction in vivo, administration of 5 mg/kg of water and methanolic extracts at day 18 showed significant anthelmintic activity with maximum reduction 86.07% and 44.15% in faecal egg counts (Lone et al., 2012). Similarly, the methanolic extract of whole plants of E. helioscopia showed antileishmanial activity against Leishmania amazonensis with IC50 of <12.5 μg/mL (Al-Sokari et al., 2015).
Euphorbia helioscopia L. which is called “Zeqi” in China, is a medicinal plant extensively distributed worldwide, especially in China, and has been widely used for decades to treat various diseases including edema, phlegm and cough, malaria, dysentery, scab, tuberculous fistula, osteomyelitis, and cancer. The present review aims to provide up-to-date information on E. helioscopia, including its traditional uses, phytochemistry, quality control, pharmacological research, toxicology, and human studies in exploring future scientific and therapeutic potentials in disease treatment. So far, a total of 173 terpenoids, as well as polyphenols, steroids, lipids, and volatile oils have been isolated and identified from E. helioscopia. Among them, diterpenoids and ﬂavonoids are the most important and abundant bioactive constituents. Modern pharmacological studies have demonstrated that E. helioscopia has outstanding bioactivities especially on antiproliferative and multidrug resistance modulating. Nevertheless, most of these studies were carried out in vitro. The quality control, metabolites identification, in-depth in-vivo studies as well as toxicology and human studies for the crude extracts and active components are still very limited. Consequently, more well-designed pre-clinical and clinical studies are required to justify their reported therapeutic potential.
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Furthermore, the NANPDB hit Euphohelionon, which is a constituent of the Euphorbia helioscopia L. plant [96] was predicted to have an anthelminthic activity with Pa of 0.378 and Pi of 0.02. A study of the methanolic extracts of E. helioscopia, the parent plant of euphohelionon, revealed significant larvicidal and ovicidal effects against H. contortus worms, both in vitro and in vivo [97]. Table 4 summarizes the PASS anthelmintic predictions of the 19 hit compounds from AfroDB [28] and NANPDB [27].
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¹: Microbiology Lab., Centre of Research for Development, University of Kashmir, Srinagar 190006, India.

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In vitro and in vivo anthelmintic activity of Euphorbia helioscopia L.

Abstract

Section snippets

Collection of plant materials

Faecal egg count reduction test

Discussion

Conclusion

Acknowledgements

Small Rumin. Res.

Vet. Parasitol.

Vet. Parasitol.

Vet. Parasitol.

Rev. Vet. Parasitol.

Exp. Parasitol.

J. Ethnopharmacol.

Vet. Parasitol.

Pakistan J. Ethnopharmacol.

J. Ethnopharmacol.

Small Rumin. Res.

Vet. Parasitol.

Toxic

Vet. Parasitol.

Acta Trop.

Int. J. Parasitol.

Vet. Parasitol.