INTERNATIONAL JOURNAL OF AGRICULTURE & BIOLOGY
In Vitro Anthelmintic Activity of Allium sativum, Zingiber
officinale, Curcurbita mexicana and Ficus religiosa
ZAFAR IQBAL, QAZI KHALID NADEEM, M.N. KHAN, M.S. AKHTAR† AND FAISAL NOUMAN WARAICH‡
Departments of Veterinary Parasitology, and †Physiology and Pharmacology, University of Agriculture, Faisalabad–38040,
Pakistan; ‡Section of Clinical Medicine and Surgery, College of Veterinary Sciences, Lahore–Pakistan
Methanol extracts of some commonly used plant materials of ethnoveterinary importance in Pakistan were screened for their in vitro
anthelmintic activity. Results revealed that Zingiber officinale killed all the test worms (Haemonchus contortus) within two hours post
exposure being 100% effective. Allium sativum and Cucurbita mexicana extracts were equally effective at 2 and 4 h post exposure; by 6 h
post exposure, however, the earlier was 100% effective; whereas, C. mexicana could not kill all the worms and was 83.4% effective. Ficus
religiosa was 100% effective by 4 h post exposure, and was as good as A. sativum and Z. officinale by 6 h post exposure. Majority of the
worms exposed to control (normal saline) remained alive till 4 h post exposure, and thereafter, 50% of them died by 6 h post exposure. It was
concluded that all the studied plants had some anthelmintic activity, therefore, in vivo trials may be conducted for further evidence for their
use in animals on scientific basis.
Key Words: Anthelmintic activity; Haemonchus contortus; Allium sativum; Zingiber officinale; Ficus religiosa; Cucurbita mexicana
Table I. Plant materials evaluated for their anthelmintic
Development of anthelmintic resistance in helminths
reported in a number of countries (Jackson, 1993; Sanyal,
1996; Rolfe, 1997; van Wyk et al., 1997; Waller, 1997)
gives a clear indication that control programs based
Liliaceae Bulb Garlick,
(a bulbous herb)
exclusively on their use are not sustainable. The
development of integrated programs to control helminths is
Zingiberaceae Rhizomes Ginger,
vital, but such control programs require viable alternatives
to the use of anthelmintics (Waller, 1999). Medicinal plants
have served through ages, as a constant source of
medicaments for the exposure of a variety of diseases. The
( a trailing herb)
history of herbal medicine is almost as old as human
Urticaceae Bark Pippal,
civilization. The plants are known to provide a rich source
(a large tree)
of botanical anthelmintics, antibacterials and insecticides
(Satyavati et al., 1976; Lewis & Elvin-Lewis, 1977). A
number of medicinal plants have been used to treat parasitic
infections in man and animals (Nadkarni, 1954; Chopra et
apparatus. The extracts thus obtained were placed in small
al., 1956, 1958; Said, 1969; Akhtar, 2000). The
test tubes separately and stored in a refrigerator for further
study was conducted to evaluate the in vitro anthelmintic
activity of some palnt materials commonly used in
Evaluation of in vitro anthelmintic activity. In vitro
ethnoveterinary medicine in Pakistan.
anthelmintic activity of the plant materials was evaluated by
exposing the adult Haemonchus contortus to undiluted
MATERIALS AND METHODS
extracts of plants. For each extract, five petridishes were
used i.e. four for extract to be tested and one for normal
Preparation of extracts. Each material (Table I) was
saline as control. Adult motile H. contortus were collected
thoroughly cleaned with water and dried under shade for
from the gastrointestinal tract of sheep slaughtered at
five to seven days until it became grindable. The dried
Faisalabad slaughterhouse and immediately transferred to
materials were ground with the help of electric grinder and
the petridishes containing plant extracts and normal saline.
separately preserved for further use. About 200 g of each
Observations were made on the motility/survival of worms
plant material was used for extraction. Each plant material
at 0, 2, 4 and 6 h post-exposure (PE).
was boiled for 24 h in 500 mL methanol in Soxhlet’s
The number of worms dead at 6 h PE to various plant
extracts was compared statistically with those exposed to
ANTHELMINTIC ACTIVITY OF SOME PLANT MATERIALS / Int. J. Agri. Biol., Vol. 3, No. 4, 2001
control group by z-test using Microsoft Excel 2000
been reported to be effective in the exposure of dysentery
and also act as vermifuge (Nadkarni, 1976; Schavenberg
& Paris, 1977). Oil of A. sativum has also been reported to
RESULTS AND DISCUSSION
possess anthelmintic activity (Steenis-Kruseman, 1953;
Hoppe, 1975; Nadkarni, 1976; Perry, 1980; Kirtikar &
It is evident from Table II that Zingiber officinale
Basu, 1981) and discards all injurious parasites in the
killed all the worms within two hours PE being 100%
intestine (Nadkarni, 1976). Garlic is the best known source
effective. Allium sativum and Cucurbita mexicana extracts
of selenium. The sulfur compound allicin, produced by
were equally effective at 2 and 4 h PE; however, by 6 h PE,
crushing or chewing fresh garlic, in turn produces other
the earlier was 100% effective; whereas, C. mexicana could
sulfur compounds: ajoene, allyl sulfides, and vinyldithiins
not kill all the worms and was found 83.4% effective, which
(Koch & Lawson, 1996).
was not different (P < 0.05) from control. Ficus religiosa
Zingiber officinale. Ginger is a perennial plant and is
was 100% effective by 4 h PE, and was as good as A.
considered to be the universal medicine in Ayurveda.
sativum and Z. officinale by 6 h PE. Majority of the worms
Traditional Chinese medicine has recommended ginger
exposed to control (normal saline) remained alive till 4 h
for over 2,500 years. It is used for abdominal bloating,
PE; therefater, three out of six worms were found dead. The
coughing, vomiting, diarrhea and rheumatism. Classified
justification as to why the aforesaid plant materials exerted
as an aromatic bitter, it stimulates digestion. It also keeps
lethal effect on H. contortus is not known by now.
the intestinal muscles toned (Bradley, 1992).
However, following information on the use, chemical
The anthelmintic activity of alcoholic extracts of
composition and available evidence suggest detailed studies
rhizomes of Z. officinale against human Ascaris
to generate more precise knowledge about these materials.
lumbricoldes is appreciable (Kalesaraj, 1974; 1975). Goto et
Allium sativum. Garlic is mentioned in the Bible and the
al. (1990) reported lethal effect of Z. officinale on Anisakis
Talmud. Its historical or traditional use (may or may not
larvae in vitro. The antifilarial effect of Z. officinale against
be supported by scientific studies) has been mentioned by
Dirofilaria immitis has been reported by Datta and Sukul
Hippocrates, Galen, Pliny the Elder, and Dioscorides for
(1987). Adeewunmi et al. (1990) have reported
many conditions, including parasites, respiratory molluscicidal and antischistosomal activities of Z. officinale.
problems, poor digestion, and low energy. Its use in China
The dried rhizome of ginger contains approximately 1–4%
was first mentioned in A.D. 510. Louis Pasteur confirmed
volatile oils. These are the medically active constituents of
the antibacterial action of garlic in 1858. It is mildly
ginger, and they are also responsible for ginger’s
antihypertensive (Silagy & Neil, 1994) and has
characteristic odor and taste. The aromatic principles
antioxidant activity (Kleijnen, 1989). Garlic has include zingiberene and bisabolene, while the pungent
antibacterial, antiviral, and anti-fungal activity (Hughes &
principles are known as gingerols and shogaols (Tyler,
Lawson, 1991). The fresh bulbs are also cooked and eaten
1994). There is speculation that the mechanism of action of
to treat asthma, colds, coughs, and to control worms. The
ginger may be both central and peripheral i.e.
alcoholic extract of bulb of A. sativum has also shown
anticholinergic and antihistaminic (Quian & Liu, 1992).
moderate in vitro anthelmintic activity against human
Cucurbita mexicana. The aqueous, etheral and alcoholic
Ascaris lumbricoldes (Kalesaraj, 1975). A. sativum has
extracts of C. mexicana seeds have exhibited good
Table II. Effect of methanol extracts of different plants on the motility/survival of Haemonchus contortus
Botanical name of plant
Time post exposure (hours)
Efficacy (at 6 hours
0 2 4 6
Allium sativum (Garlick)
Zingiber officinale (Ginger)
Cucurbita mexicana (Kaddu)
Ficus religiosa (Pipal)
Normal saline (control)
* indicates significant (P < 0.05) difference compared with control independently; NS = Non-significant
IQBAL et al. / Int. J. Agri. Biol., Vol. 3, No. 4, 2001
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