Pharmacologyonline 1: 524-527 (2011) ewsletter Shah et al.

I VITRO A THELMI TIC ACTIVITY OF LEAF EXTRACT OF AVERRHOA

CARAMBOLA AGAI ST PHERETIMA POSTHUMA

Anisha . Shah*, Anuja B. Raut*, Prof. Akshay Baheti, Dr. B.S. Kuchekar

MAEER’s Maharashtra Institute of Pharmacy, Pune-38, Maharshtra, India.

Address correspondence to:

Miss. Anuja B. Raut
MAEER’s Maharashtra Institute of Pharmacy,
Pune, Maharashtra, India.
Pin- 411038.
Phone: +91 9561709340
Email address: [email protected]

Summary
The aim of the present study was to evaluate the anthelmintic potential of Averrhoa
carambola using Pheretima posthuma as the test worm. Various concentrations (10-
100mg/ml) of the drug were tested in the bioassay, which involved determination of
time of paralysis (P) and time of death (D) of the worms. Albendazole (10-100mg/ml)
was included as standard reference and distilled water as control. The result of the
present study indicated that Averrhoa carambola significantly demonstrated paralysis,
and also causes death of worms especially at higher concentration of 100mg/ml, as
compared to standard reference Albendazole. In conclusion the anthelmintic activity of
Averrhoa carambola has been confirmed.
Key Words: Averrhoa carambola, Pheretima posthuma, Albendazole.

Introduction
Soil-transmitted helminth (STH) infections are among the most prevalent of chronic
human infections worldwide1. Helminthes are generally restricted to tropical regions and
cause serious problems to health and contribute to the prevalence of undernourishment,
anemia, eosinophilia and pneumonia2. After decades of serious obsession with the
modern medicinal system, people have started looking at the ancient healing systems
like Ayurveda, Siddha and Unani3. The WHO has estimated that 80% of the world’s
population already relies on traditional medicine for their primary healthcare needs,
therefore traditional medical plants being an important component of complementary
and alternative medicine may be useful for drug discovery and development4.
Averrhoa carambola, traditionally known as ‘karmakh’, and commonly known as star
fruit because of its peculiar shape, belongs to the plant family Oxalidaceae, has been
widely used in Ayurveda, preparations of its fruit and leaves are used to pacify impaired
kapha, pitta, skin diseases, pruritis, worm infestations, diarrhea, vomiting, hemorrhoids,
intermittent fever, over-perspiration and general debility. It is also used in traditional
medicines in countries like China, Phillipines, Brazil for various ailments3.

524
Pharmacologyonline 1: 524-527 (2011) ewsletter Shah et al.
Recent studies show that Averrhoa carambola is a very rich source of natural
antioxitants5, studies have also proven hypoglycaemic effects6, antimicrobial propery7of
this plant.
The plant is believed to have originated in Ceylon and is commonly grown in the
provinces of Fukien, Kuangtung and Kuangsi in southern China, in Taiwan and India. It
is rather popular in the Phillipines and Queensland, Australia and is widely used as an
ornamental plant and in pickles and jams8.
Averrhoa carambola is a small, evergreen, multistemmed tree, 3-5m high or, rarely,
10m high. At the base, the trunk reaches a diameter of 15cm, the bark is light brown,
smooth or finely fissured. Leaves are 15-25cm long alternate, pinnate, disposed more or
less in a horizontal plane, shortly petiolate with 7-9 pendant leaflets; leaves have the
peculiarity of being sensitive to touch in the same way as certain Mimosa species [8, 9].
Preliminary phytochemical analysis indicated presence of saponins, tannins, alkaloids
and flavonoids3 hinting the presence of anthelmintic activity, but as there are no
scientific evidences to support the same, the aim of this study was to test the
anthelmintic activity of the leaf extract of Averrhoa carambola on adult Indian earth
worms (Pheretima posthuma).
Materials and Methods
Plant material
Averrhoa carambola leaves were collected by A.N. Shah from Pune region in December
2010 and identified by Pharmacognosy department of MAEER’s Maharashtra Institute
of Pharmacy, Pune.
Worm collection
Healthy adult Indian earthworm Pheretima posthuma about 5-7 cm were purchased from
Enora Pvt. Ltd, Pune, washed and kept in purified water.
Preparation extract
The leaves of Averrhoa carambola were dried and ground to coarse powder and
macerated for 72 hours. The aqueous extract obtained was filtered and concentrated by
evaporating to get extract.
Preparation of test sample
Samples for in-vitro study were prepared by dissolving 2.5gms of extract in 25ml of
distilled water to obtain stock solution of 100mg/ml .From this stock solution different
working dilutions were prepared to get a concentration range of 10, 50 and 100mg/ml.
Anthelmintic Assay
The anthelmintic assay was carried out as per Ajaiyeobaet al9, with minor modifications.
The assay was performed on Indian earthworm Pheretima posthuma due to its
anatomical and physiological resemblance with intestinal roundworm parasite of human
beings[10-12]. Because of easy availability, earthworms have been used widely for the
initial evaluation of anthelmintic compounds in-vitro[13-15]. 50 ml formulations
containing three different concentrations each of extract (10, 50, 100 mg/ml in distilled
water) were prepared and six worms (same type) were placed in it. Time for paralysis
was noted when no movement of any sort could be observed except when worms were
shaken vigorously. Time of death of worms was recorded after ascertaining that the
worms neither moved when shaken vigorously or when dipped in water (500C).
Albendazole was used as reference standard (10, 50, 100 mg/ml) while distilled water as
the control.

525
Pharmacologyonline 1: 524-527 (2011) ewsletter Shah et al.

Results and Discussion

Preliminary phytochemical analysis indicated presence of saponins, tannins, alkaloids

and flavonoids in aqueous extract of Averrhoa carambola. The leaves of Averrhoa
carambola displayed a significant anthelmintic activity in dose depended manner giving
shortest time of paralysis (P) and death (D) with100mg/ml concentration as shown in
Table 1. The anthelmintic activity of aqueous extract was comparable with that of
standard drug at 100mg/ml concentration, plant extract showed paralysis in 10 minutes
and death in 16 minutes while Albendazole exhibited similar effects at 10 and 21
minutes respectively. As the plant extracts are found to contain tannins, which are
polyphenolic compounds, there is a possibility that their effects are like synthetic
phenolic anthelmintics e.g.- niclosamide, oxyclozanide, bithionol etc. which are reported
to interfere with energy generation in helminth parasites by uncoupling oxidative
phosphorylation16. Another possible anthelmintic activity of tannins is that they can bind
to free proteins in gastrointestinal tract of host animal17 or glycoprotein on the cuticle of
parasite18 and may cause death.

Table o. 1: Anthelmintic activity of Averrhoa carambola

Time taken for Paralysis (P) and

Sample Concentration Death (D) of Pheretima posthuma
(mg/ml) in minutes
P D
Albendazole 10 31.16±2.48 72.16±8.47
Albendazole 50 14.16±1.16 47.5±1.22
Albendazole 100 10.5±1.04 21.33±1.63
Plant extract 10 87±13.57 179.5±24.22
Plant extract 50 26.83±3.71 35±2.82
Plant extract 100 10.83±1.72 16.16±1.85
All values represent Mean ± SEM; n=6 in each group. Control worms were alive up to
24 hours.

Conclusion

Averrhoa carambola showed anthelmintic activity. However dose and form in which
they can be used requires standardization. Future scope involves isolation of
phytoconstituents responsible for activity.

Acknowledgement

The authors are thankful to Dr. B.S Kuchekar, Principal MAEER’s Maharashtra Institute
of Pharmacy, Pune for providing necessary facilities and co-operation during this
research work.

526
Pharmacologyonline 1: 524-527 (2011) ewsletter Shah et al.
References

1] Brooker S, Clements ACA & Bund DAP, Global Epidemology Ecology and Control Of Soil
Transmitted Helminth Infections, Advances in Parasitology 2006; 62: 221-261.
2] Bundy DA, Immunoepidemiology of intestinal helminthic infection: The global burden of
intestinal nematode disease, Trans Royal Soc Trop Med Hyg 1994; 8: 259-261.
3] Thomas S., Patil D.A., Pharmacognostic Evaluation and Physicochemical Analysis of
Averrhoa Carambola L., Fruit Journal of Herbal Medicine and Toxicology 2008; 2 : 51-54.
4] Deore SL, Khadbadi SS., In vitro anthelmintic studies of Chlorophytum borivilianum sant. &
Fernandez tubers, Indian Journal of Natural Products and Resources 2010;1:53-5.
5] L.P. Leong, G. Shui., Residue from star fruit as valuable source for functional food
ingredients and antioxidant nutraceuticals, Food Chemistry 2006; 97: 277-284.
6] Chi-Fai Chau, Chien-Hung Chen, Ching-Yi Lin, Insoluble fiber-rich fractions derived
from Averrhoa carambola: hypoglycemic effects determined by in vitro methods .Lebensmittel-
Wissenschaft und-Technologie 2004; 37 (3,suppl) : 331-335.
7]Mohamed,Suhaila and Hassan, Zahariah and Abd Hamid, Norhashimah Antimicrobial Activity
of some Tropical Fruit Wastes (Guava, Starfruit, Banana, Papaya, Passionfruit, Langsat, Duku,
Rambutan and Rambai. Pertanika Journal of Tropical Agricultural Science 1997, 17
(3,suppl):219-227.
8] http://www.hort.purdue.edu/newcrop/morton/carambola.html
9]Ajaiyeoba EO, Onocha PA, Olarenwaju OT. In vitro anthelmintic properties of Buchholzia
coriaceae and Gynandropsis gynandra extract. Pharm Biol. 2001; 39: 217.
10]Chatterjee K.D. Parasitology, Protozoology and Helminthology, Guha Ray Sree Saraswaty
Press Ltd., Calcutta, 1967:168–169.
11]Vidyarthi R.D., A Text Book of Zoology, S.Chand and Co., New Delhi, 1967.
12]Thorn G.W., Adams R.D., Braunwald E., Isselbacher K.J. and Petersdorf R.G., Harrison’s
Principles of Internal Medicine, McGraw Hill Co., New York, 1977: 1088.
13]Baheti A M, Khandelwal K. R., Baheti Shikha A. In vitro evaluation of anthelmintic activity
of anterdhumpadhatimashi and bahirdhumpadhatimashi of green Cocosnucifera Linn. (Palmae)
husk. Phcog.Net2009; 1(4,suppl):279-281.
14]Sollmann T. Anthelmintics: Their efficiency as tested on earthworms. J.Pharmacol.Exp.Ther
1918; 12: 129–170.
15]Ganapaty Dash G.K., Suresh P., Sahu S.K., Kar D. M. S. and Panda S.B. Evaluation of
Evolvulusalsinoids Linn. for anthelmintic and antimicrobial activities. J.Nat.Rem2002; 2:182–
185 .
16]Chung Ki Sung, Takeatsu Kimura, Paul P. H. But, Ji-Xian Guo. International Collation of
Traditional and Folk Medicine: Northeast Asia. World Scientific Publishing.
16]Martin R.J. Mode of action of anthelmintic drugs. Vet J 1997; 154:11-34.
17]Anthnasiadou S., Kyriazakis F.,Jackson R.L., Direct anthelmintic effect of condensed tannins
towards different gastrointestinal nematodes of sheep: In vitro and in vivo studies. Vet. Parasitol
2001; 99:205-219.
18]Thompson D.P.,Geary T.G.The structure and function of helminth surfaces. In: Marr, J. J.
eds. Biochemistry and Molecular Biology of Parasites. Academic Press, New York, 1995: 203-
232.

527