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In-vitro anthelmintic activity of fenugreek leaves (aqueous extract) in Indian earthworms

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In-vitro anthelmintic activity of fenugreek leaves (aqueous extract) in Indian earthworms

Abstract

Background-Soil transmitted helminthiasis (STH) is one of the common health problems of under developed countries. Fenugreek is a popular food substitute in Indian kitchen and ayurveda for centuries. Even though a number of the anthelmintic drugs available in the market with considerable side effect, we studied in vitro anthelmintic activity of fenugreek leaves extract in comparison with Albendazole. Methods-This comparative experimental study was done over the period of 3 weeks in Indian adult earthworm due to its anatomical resemblance with the Ascaris lumbricoides. An aqueous extract is prepared from standard method. Albendazole was used as comparator drug; 20 mg/ml concentration was prepared as per the approved method. Each group with six earthworms of equal was released into in each 50 ml of desired concentration of comparator and fenugreek leaves extract in the petridish. 50 ml contained 200 mg/ml of aqueous extract of leaves of fenugreek. Earth worms in the petridish were studied for paralysis /death as end points of our study. The mean time to paralysis/death was recorded and expressed in minutes. The test results were compared with comparator. Results-Aqueous extract of fenugreek leaves was 144±15 minutes to paralyze and 224±10 minutes to death of the worm, whereas albendazole took 90±12 minutes to paralyze and 155±21 minutes to death of the worms is statistically significant (P<0.05). Fenugreek leaves aqueous extract is more potent than control (NS) and lesser anthelmintic activity than Albendazole. Conclusion-Aqueous extract fenugreek leaves showed noteworthy anthelmintic activity as compared to control (NS) but less potent than albendazole.
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The Pharma Innovation Journal 2016; 5(4): 70-72
ISSN: 2277- 7695
TPI 2016; 5(4): 70-72
© 2016 TPI
www.thepharmajournal.com
Received: 10-02-2016
Accepted: 13-03-2016
Dr Madhavulu Buchineni
Associate Professor,
Dept. of Pharmacology
Narayana Medical College
Nellore – 524 003
Andhra Pradesh.
Dr Souris Kondaveti
Assistant Professor
Dept. of Pharmacology
Osmania Medical College
Hyderabad
Corresponding Author
Dr Madhavulu Buchineni
Associate Professor,
Dept. of Pharmacology
Narayana Medical College
Nellore – 524 003
Andhra Pradesh
Email: madhavulu@gmail.com
Mob: 9440 713718
In-vitro anthelmintic activity of fenugreek leaves
(aqueous extract) in Indian earthworms
Madhavulu Buchineni, Souris Kondaveti
Abstract
Background- Soil transmitted helminthiasis (STH) is one of the common health problems of under
developed countries. Fenugreek is a popular food substitute in Indian kitchen and ayurveda for centuries.
Even though a number of the anthelmintic drugs available in the market with considerable side effect, we
studied in vitro anthelmintic activity of fenugreek leaves extract in comparison with Albendazole.
Methods- This comparative experimental study was done over the period of 3 weeks in Indian adult
earthworm due to its anatomical resemblance with the Ascaris lumbricoides. An aqueous extract is
prepared from standard method. Albendazole was used as comparator drug; 20 mg/ml concentration was
prepared as per the approved method. Each group with six earthworms of equal was released into in each
50 ml of desired concentration of comparator and fenugreek leaves extract in the petridish. 50 ml
contained 200 mg/ml of aqueous extract of leaves of fenugreek. Earth worms in the petridish were
studied for paralysis /death as end points of our study. The mean time to paralysis/death was recorded
and expressed in minutes. The test results were compared with comparator.
Results- Aqueous extract of fenugreek leaves was 144±15 minutes to paralyze and 224±10 minutes to
death of the worm, whereas albendazole took 90±12 minutes to paralyze and 155±21 minutes to death of
the worms is statistically significant (P<0.05). Fenugreek leaves aqueous extract is more potent than
control (NS) and lesser anthelmintic activity than Albendazole.
Conclusion- Aqueous extract fenugreek leaves showed noteworthy anthelmintic activity as compared to
control (NS) but less potent than albendazole.
Keywords: Soil Transmitted Helminthiasis (STH), Fenugreek leaves aqueous extract, Albendazole,
Normal Saline (NS)
Introduction
Soil transmitted helminthiasis (STH) are one of the common health problems affecting
developing and under developed countries
[1]
. Several studies have suggested that mild to
moderate intensity of infection may result in deferred physical growth, impaired cognitive
functions, absenteeism among school going children
[2-4]
. Antihelminthics are the agents that
expel helminthes from the body either stunning or killing them
[5]
. There are few plants known
to provide a rich source of phyto-anthelmintics in the nature
[6-7]
. Trigonella foenum-graecum
belongs to the family Leguminoceae and it is popularly known as Fenugreek (Methi) in India
[8]
. It is also stated in Ayurveda and Siddha (Indian traditional systems of medicine) that these
plants were used to treat different kinds of fever, dysentery and heart diseases, whereas in
Unani system, this plant is used as an aphrodisiac, diuretic
[9-10]
. To this purpose we studied in
vitro anthelmintic activity of fenugreek leaves extract in comparison with albendazole.
Methods
This comparative study was done at Pondicherry institute of medical sciences, Puducherry
over the period of 3 weeks during in the month of January 2011. The experiment was done in
Indian earthworms (Pheretima posthuma) adult type due to their anatomical resemblance with
the Ascaris lumbricoides. The Institute Ethical Committee has approved the study protocol.
The earthworms were obtained from moist soil and cleaned with water to remove all dirt.
Fresh fenugreek leaves were purchased from the local market then, it was endorsed by
certified Taxonomist from Pondicherry University, Puducherry.
The leaves dried under shade and crushed in an electric blender to form powder and subjected
Soxhlet
extraction
using
water
as
solvent.
Aqueous
extract
is
prepared
from
standard
methods
[11]
. Albendazole was used as comparator drug; 20 mg/ml concentration was prepared by as per
the approved method. The suspension of aqueous extract of leaves of fenugreek concentration
200 mg/ml was prepared and final volume was made up to 50 ml for relevant concentration.
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The Pharma Innovation Journal
Groups of equal size worms consisting of six earthworms
individually in each group were released into in each 50 ml of
desired concentration of drug and extracts in the petridish. The
anthelmintic activity was performed according to standard
screening methods [12-14]. Six Indian adult earth worms were
placed in petridish containing 50 ml contained 200 mg/ml of
aqueous extract of leaves of fenugreek. Earth worm in the
petridish was studied for paralysis/death as an end point of our
study. The mean time to paralysis/death was recorded and
expressed in minutes. The test results were compared with
albendazole (20 mg/ml). The procedure continued for 3 times
to verify the results.
Statistical Analysis: Mean, SD and percentages were used to
describe the results data. Chi-square and unpaired “t” test were
used appropriately as inferential tools. P value <0.0.5 was
considered statistically significant.
Results
Aqueous extract of fenugreek leaves was 144±15 minutes to
paralyze and 224±10 minutes to death of the worm, whereas
Albendazole took 90±12 minutes to paralyze and 155±21
minutes to death of the worm with significant (P<0.05) value.
Aqueous extract is more potent than control (NS) and lesser
anthelmintic activity than Albendazole. Time to paralysis and
consequent death were significantly higher in aqueous extract
of fenugreek that of Albendazole at same concentrations.
Table 1: Antihelmintic activity of fenugreek leaves
S.
No Antihelmenthic
Substance Time taken for
Paralysis (Minutes) Time taken for
Death (Minutes)
1 Normal Saline No action took No action
2 Aqueous extract
of fenugreek 144±15 224±10
3 Albendazole 90±12 155±21
P-Value P<0.05 P<0.05
Discussion
Soil transmitted helmintic infestation is wide spread in rural
India. Fenugreek leaves are cheap, easily available in the local
market, therefore this can be tried as an alternative remedy to
already existing standard anthelmintic drugs. A number of
medicinal plants have been used to treat parasitic infections in
humans and animals [15]. Anthelmintic derived from plant
source can be an answer to this STH problem as they form
secure and non-toxic with a modified site of action [16-17].
Chandrashekhar et al. [18] and Bhalke et al. [19] in their study
found that seeds and leaves of Trigonella Foenum-graecum
showed marked anthelmintic activity.
Limitations of the Study
The bioactivity guided demonstration of anthelmintic activity
helps in standardization of its anthelmintic activity.
Conclusion
Aqueous extract fenugreek leaves showed noteworthy
anthelmintic activity as compared to normal saline but less
potent than albendazole. Additional large studies are warranted
using in vivo models to prove fenugreek leaves as better
anthelmintic substance.
Funding: None
Conflict of Interest: None declared
Ethical Approval: None required
References
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19. Bhalke RD, Anarthe SJ, Sasane KD, Satpute SN, Shinde
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87
Chapter
Trigonella foenum-graecum L. (fenugreek) is cultivated in various countries and consumed for many goals including medicinal applications (anti-diabetic, lowering cholesterol level, antioxidant, anti-microbial, anti-ulcer, lactation aid, and anti-obesity), making food (like stew with rice), roasted grains coffee-substitute, and controlling insects in grain storages. Generally, the seeds and leaves of the crop are commonly applied in food and pharmaceutical industries and contain several active ingredients that is linked to the properties of this plant. The chemical composition of fenugreek crop includes moisture, ash, protein, mucilage, lipids, alkaloids (mainly trigonelline), amino acids (particularly 4-hydroxyisoleucine), saponins (mainly diosgenin), flavonoids (mainly quercetin, rutin, and vitexin), fibers (mainly soluble dietary fiber), polyphenols, coumarin, vitamins (mainly vitamin C), and minerals (mainly calcium, zinc, sulfur, and phosphorus). Fenugreek as a hydrocolloid (known as galactomannan) can be applied as a thickening, emulsifying, stabilizing, gelling, and encapsulating agents and also it has positive effects on the textural and appeal properties of food and pharmaceutical products. In this chapter, a review of the methods for extraction and identification of different substances present in fenugreek is evaluated.
Chapter
Fenugreek is an herb which has been used in traditional medicines for centuries in wound healing, as an aphrodisiac, for promotion of lactation, etc. The consumption of the seeds as a spice results in different medicinal effects such as hypocholesterolemic, antidiabetic, hepatoprotective, antibacterial, anthelmintic, anticancer, and galactogogue. Flavonoids, saponins, pyridine alkaloids, and steroidal sapogenins are some of the phytochemicals present in the plant. The plant is also embraced for its high content of important vitamins, minerals, protein and amino acids, and fibers making it a nutritious fodder for livestock. Extracts of the leaves and seeds of fenugreek are considered safe and are found to have potential therapeutic explicabilities in the treatment and/or management of diabetes, cancer, toxicities, cardiovascular diseases, physical injuries, and hormonal imbalances. The seeds and leaves of this plant are now being incorporated into animal, bird, and fish foods to increase feed intake, to promote weight gain, and to decrease the feed conversion ratio. The addition of fenugreek in the drinking water of poultry reduces stress, and this can be an important strategy to replace the use of antibiotics such as enrofloxacin as an anti-stress agent, and thus the issues of antibiotic residues in meat, as well as widely developing antibiotic resistance, would be less.
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