Anthelmintic activity of Cassia auriculata L. extracts-In vitro study

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J. Nat. Prod. Plant Resour., 2011, 1 (2): 62-66
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ISSN : 2231 – 3184
CODEN (USA): JNPPB7
Anthelmintic activity of Cassia auriculata L. extracts-In vitro study
Sucheta A Gaikwad*, Asha A. Kale, Balaso. G. Jadhav, Nirmala. R. Deshpande and Jyoti.
P. Salvekar
Dr. T. R. Ingle Research Laboratory, Department of Chemistry, S. P. College, Pune
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ABSTRACT
Helminth infections are among the most common infections in humans, affecting a large
population of the World. Use of herbal products as antimicrobial agents may provide the best
alternative to the wide and injudicious use of synthetic antibiotics. Albendazole, the commercial
anthelmintic drug is used as a standard reference and saline as control. As per WHO, only few
drugs are frequently used in the treatment of these parasite infections In present study acetone,
ethanol, methanol and quath extracts of Cassia auriculata L leaves were investigated for their
anthelmintic activity against Ecinia foeitida. Three concentrations (5, 10, 20 mg/ml) of each
extract were studied. This experiment involved the determination of time of paralysis and time of
death of the worms. The gradual increased in a dose exhibited, a gradual increase in the activity.
Quath and ethanol extracts displayed significant anthelmintic activity at highest concentration of
20 mg/ml. It was concluded from the experimental detail study that the plant revealed significant
anthelmintic activity, therefore in vivo trials may be conducted for the use in livestock on
scientific basis. The data were verified as statistically significant by using two way ANOVA at
1% level of significance (P-value < 0.01)
Keywords: C. auriculata L, anthelmintic activity, Ecinia foeitida, leaves extracts.
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INTRODUCTION
Helminthiasis or worm infection is one of the most prevalent diseases in the World. The
conditions appear to be worsening with respect to the number of persons affected[1]. The WHO
estimates that a staggering two billion people harbor parasitic worm infections. Parasitic worm
also infect livestock and crops, affecting food production with a resultant economic impact.
Despite this prevalence of parasitic infections, the research on anthelmintic drug is poor[2].
Helminth infections are among the most common infections in humans, affecting a large
population of the World Although the majority of infections due to worms are generally limited
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to tropical regions and pose a great threat to health and contribute to the prevalence of
malnutrition, anaemia, eosinophilia and pneumonia[3].Parasitic diseases cause severe morbidity
affecting mainly population in endemic areas with major economic and sol consequences[4]. The
gastro-intestinal helminthes becomes resistant to currently available anthelmintic drugs therefore
there is a foremost problem in treatment of helminthes diseases[5]. There is an increasing
demand towards natural anthelmintics. The plant C.auriculata L. is an aromatic more or less
deciduous shrub found through out India up to an altitude of 1,500 m commonly in forests. C.
auriculata L. (Caselpinaceae) is a tall, much branched bushy shrub growing wild throughout
forests and along roadsides and in wastelands. Literature survey reveals that leaves of C.
auriculata L are useful in treatment of leprosy, ulcers and have anthelmintic activity[6]. The
anthelmintic activity of quath extract of the leaves is reported[7]. The plant is reported for the
presence of polyphenolic compounds.anthelmintic activity, chemically tannins are polyphenolic
compounds[8]. Some synthetic phenolic anthelmintics e.g. niclosamide, oxyclozanide and
bithionol are shown to interfere with energy generation in helminth parasites by uncoupling
oxidative phosphorylation[9]. Another possible anthelmintic effect of tannins is that they can
bind to free protein in the gastrointestinal tract of host animal or glycoprotein on the cuticle of
the parasite and cause death[10].The present study investigates the anthelmintic activity of the
leaves for the different extracts with a view to justify the use of the plant in the treatment of
helminths.
MATERIALS AND METHODS
1. Plant materials and preparation of extracts
Cassia auriculata L was collected from Western Pune, Maharashtra, India. The taxonomic
identification was carried out with the help of Flora of Botany Presidency and Flora of
Maharashtra (Singh et al.2000) and herbaria were prepared by following standard methods. The
specimens were also compared with the authentic herbaria of BSI, Western circle
Pune,Maharashtra,India for confirming the identification, its number is BSI/WC/Tech/2009/95.
Air shade dried and pulverized plant material was used. Extracts were prepared using exact
weighed sample powder in the measured volume of solvents like, acetone, ethanol, methanol
and quath.Vaccum dried extracs are used for the experiment.Solvents used after distillation.
2. Phytochemical screening
Qualitative assay, for the presence of plant phytoconstituents such as carbohydrates, alkaloids,
glycosides, flavonoids, tannins and saponins were carried out on following standard procedure
[7,11].
3.Collection of Animal
The anthelmintic assay was carried as per the method of Ajaiyeoba et. al. [9,14] with minor
modifications[12,15]. The assay was performed on adult earthworm (Eicinia fetida) owing to its
anatomical and physiological resemblance with the intestinal roundworm parasites of human
beings[13,14]. Easy availability of earthworms prompts their extensive use for preliminary in
vitro evaluation of anthelmintic compounds[10]. Earthworms were provided and identified by
Dept. of Entomology, College of Agriculture, Pune,Maharashtra,India. All earthworms were of
approximately equal size (15 cm). They were washed with saline before experiment.
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4. Drugs
Albendazole was purchased from G.S.K. Pharma Pvt ltd, Mumbai along with saline NA 0.91 %
NaCl . The solvents of analytical grade were used during experimental protocol.
Experimental
Quath, ethanol, methanol, ethylacetate, extracts of Plant material investigated for their
anthelmintic activity against Ecinia foeitida. The anthelmintic assay was carried as per the
method of Ajaiyeoba et. al.[9] with minor modifications. All the test solutions and standard drug
solutions were prepared freshly before starting the experiment. Six groups of earthworms of
approximately equal size were released in to 25 ml solutions of three different concentrations
(5,10,20 mg/ml) in petri dishes containing above solutions of extracts. Albendazole was used as
reference standard and saline as control. Determination of time of paralysis and time of death of
the worm were done. Time for paralysis was noted when no movement of any sort could be
observed except when the worms were shaken vigorously. Time for death of worms was
recorded after ascertaining that worms neither moved when shaken vigorously nor when dipped
in warm water (50 o C) followed with fading away of their body colours.
Ethanopharmocological Relevence
The plant species screened in this research was recorded by several indigenous medicinal
practitioners as antiparasitic, anticancer and/or related activities to the human major organ
system.
Table I Shows Anthelmintic Activity results of Cassia auriculata L. Extracts.
Eicinia foeitida (Earthworms)
Extracts Concentrations
mg/ml
Time for
paralysis(P) in min.(Mean &SEM)
Time for death (D)in min.
(Mean &SEM)
Control(Saline) ------ ----- ------
5 76±0.190 182±0.022
Quath
10 61±0.020 83±0.020
20 44±0.019 67±0.019
5 70±0.020 132±0.022
Ethanol
10 45±0.020 55±0.021
20 30±0.019 50±0.020
5 96±0.020 191±0.020
Methanol
10 85±0.020 120±0.032
20 26±0.021 45±0.019
5 85±0.019 210±0.013
Acetone
10 42±0.022 80±0.020
20 35±0.018 60±0.020
5 147±0.018 230±0.021
Std. Albendazole 10 136±0.020 195±0.020
20 1150.026 157±0.025
Values are mean ± S.E.M. from six observations .
P: Time for Paralysis (min), D: Time for Death of worms (min)
The data were verified as statistically significant by using two way ANOVA(TableII) at 1% level of
significance (P-value < 0.01)
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RESULTS AND DISCUSSION
In conclusion, this study suggests that the plants used by tribals traditionally to treat intestinal
worm infections, showed significant anthelmintic activity. The experimental evidence obtained
in the laboratory model could provide a rationale for the traditional use of these plants as
anthelmintic. The extracts of leaves displays a significant anthelmintic activity in dose
dependent manner ( TableI). The anthelmintic activity of ethanolic and quath extracts was
comparable with that of standard drug. Albendazole, this drug is effective in a broad range of
helminth infections, including round worms, hookworms, whipworms, pinworms and animal
side effects. The drug is poorly absorbed and efficacy depends on transit time in the
gastrointestinal tract. The toxicity is extremely low, but the drug has not been studied in children
under two years of age. Gastro intestinal disturbances may occur during expulsion of the worms,
the mechanism of action involves inhibition of the glucose uptake system leading to a lethal
depletion of energy reserves in the helminthes. Phytochemical screening of the crude extracts
reveales the presence of flavonoids and polyphenolic compound as one of the major chemical
constituents. It is possible that tannins contained in the extracts of Cassia auriculata L produced
similar effects.
Graph I Shows Comparative study on anthelmintic activity of Cassia auriculata L. Extracts on adult Indian
earthworm.
Time (Min)
Treatments
Table II Shows Statistical varience of analysis (ANOVA) on anthelmintic activity of Cassia auriculata L.
Extracts
Source of Variation SS df MS F P-value F crit
Between groups 14117.74 4 3529.4 5.731 0.01158 5.994
Within groups 6158 10 615.8
Total 20275.74 14
Each value represents mean ±SEM(N=6).Data are found to be significant by testing through two way ANOVA with
replication at 1% level of significance.
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CONCLUSION
It is concluded based on the findings of the present study that the leaves of Cassia auriculata L.
possess varying degree of anthelmintic activities. The dose of the extract is increased, a gradual
increase in the activity of extracts is dose dependent.(GraphI) Quath and ethanol extracts
exhibited significant anthelmintic activity at C.auriculata highest concentration of 20 mg/ml.
This study strongly supports the traditional use of leaves as anthelmintic.
Acknowledgement
The authors are thankful to the Principal, S.P.College and Head, Department of Chemistry,
S.P.College, Pune-30 for providing experimentation facilities.
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