In Vitro Evaluation of Anthelmintic activity of Rhus mysorensis leaves

 

Swathi. H^1*, T. Shekshavali²

¹Student, 2^nd year M. Pharm., Department of Pharmacology, National College of Pharmacy, Balraj Urs Road, Shivamogga 577201, Karnataka, India.

²Lecturer, Department of Pharmacology, National College of Pharmacy, Balraj Urs Road, Shivamogga 577201, Karnataka, India.

 

ABSTRACT:

The present study has been designed to elucidate the Anthelmintic activity of leaves of Rhus mysorensis using experimental models. Helminthic infections are among the most common infections in the human beings. The anthelmintic activity was carried out on adult Indian earthworm (Pheretima posthuma). All the extracts of leaves of Rhus mysorensis demonstrated a significant anthelmintic activity in causing paralysis and death of earth worms when compared to reference standard and normal saline as control.

 

 

 

1. INTRODUCTION:

Helminthic infections are among the most common infections in the human beings, affecting a large population of the world’s population. In the developing countries they pose a large threat to human and contribute to the prevalence of anemia, malnutrition, eosinophilia and pneumonia¹. The development of resistance in helminthes against conventional anthelmintics is a foremost problem in treatment of helminthes diseases^2,3. Therefore it is important to look for alternative strategies against gastrointestinal nematodes, which have led to the proposal of screening medicinal plants for their anthelmintic activity.

 

Rhus mysorensis is one among such traditionally used plants which has reported to have medicinal properties. Rhus mysorensis is Shrub with thorny branches found in hot dry places in Punjab, Haryana, Rajasthan, Gujarat, Karnataka, Andhra Pradesh and the Western. Also grow on stony regions. Reports suggest that the plant contains Alkaloids, Glycosides, Flavonoids, Saponins, Tannins, Sterols, Phenols, Amino acids, and Protein. The previous studies of the plant claims that it have potential hepatoprotective, Antimicrobial, antiurolithiatic, anti-diabetic, hypolipidemic and anti-oxidant activities. Folk usage of the plant suggested that its use in diabetes, antifertility, psoriasis and diarrhea problems⁴.

 

The aim of the present study is to evaluate the In vitro anthelmintic activity of Rhus mysorensis leaves.

 

2. MATERIALS AND METHODS:

2.1. Collection and authentication of Rhus mysorensis

The leaves of Rhus mysorensis was collected from in and around the Chitradurga district of Karnataka and was authenticated by the Botanist, Dr. Rudrappa, Department of Botany, SRNMC, Shivamogga. The leaves of Rhus mysorensis were shade dried, and reduced to a coarse powder and used for extraction.

2.2. Extraction of the plant using different solvents                                                                                            

The ethanol and petroleum ether extract of the plant material were prepared by continuous soxhlet extraction method. The powdered material of Rhus mysorensis was extracted with petroleum ether and ethyl alcohol in a soxhlet extractor for 48hrs in 3 batches. The extract was concentrated in vacuum using rotary flash evaporator (Buchi, Flawil, Switzerland). The Aqueous extract of plant material was prepared by Cold Maceration method⁵, the coarsely powdered plant material is kept in contact with the solvent in a stoppered container for a defined period with frequent agitation until soluble matter is dissolved. The solvent was removed completely over the water bath and finally desiccator dried. The extract, so obtained was used to evaluate Anthelmintic activity.

 

2.3. Phytochemical investigations of extracts 

All the extracts were screened for the presence or absence of secondary metabolites such as reducing sugars, alkaloids, steroidal compounds, phenolic compounds, tannins, saponins, flavonoids, glycosides, and anthraquinones using standard procedures^6,7.

 

2.4. Drug

Albendazole was used as standard anthelmintic agent, purchased from local medical shop of    Shivamogga, Karnataka.

 

2.5. Evaluation of Anthelmintic activity

The anthelmintic activity was carried out on adult Indian earthworm (Pheretima posthuma) according to the method of Ghosh et al. with necessary modifications. Due to the anatomical and physiological resemblance with the intestinal roundworm parasites. Samples were prepared by triturating the crude extract with distilled water using Tween 80 (1%) as suspending agent and Albendazole (standard) was prepared by diluting the drug with distilled water to a concentration of 20 mg/ml. The formulation (25ml) containing different concentration of crude extracts (50,100mg/ml) and standard (20mg/ml) were prepared and approximately equal size six earthworms were released in each group. Observations were made for the time taken to paralysis or death of individual worms. Paralysis was said to occur when the worms do not revive even in normal saline. Time for death of worms was recorded after ascertaining that when the worms stopped the movement in the suspension even after touching with a point edge of the pin without injuring the worms, and no movement observed by transferring the worms into a beaker containing hot water (50⁰C), which stimulates and induces movement if the worms are alive and followed with their body colour fading away^8,9. 

 

 

3. RESULTS:

The preliminary phytochemical analysis showed the presence of Alkaloids, Glycosides, Flavonoids, Carbohydrate, Saponins, Steroids, Tannins and Triterpenoids.

 

All the extracts of leaves of Rhus mysorensis demonstrated a significant anthelmintic activity in causing paralysis and death of earth worms when compared to reference standard and normal saline as control among which ethanol extract was found more potent. The results are shown in the Table No.1.

 

Table No.1: Effect of leaves of Rhus mysorensis extracts on the worms at different concentrations.

Groups	Concentration (mg/ml)	Time Taken for Paralysis of Worms (min)	Time Taken for Death of Worms (min)
Control	-	-	-
Standard	20	24.64±0.38	56.18±0.89
PEERM	50	38.22±0.39	144.78±0.49
	100	32.04±0.36	87.11±0.56
EERM	50	27.35±0.24	62.92±0.52
	100	25.62±0.43	59.74±0.34
AERM	50	31.57±0.34	121.26±0.56
	100	28.84±0.58	80.96±0.32

Note: Values are expressed as mean ± S.E.M. n=6 in each group. PEERM- Petroleum ether extract of Rhus mysorensis, EERM- Ethanolic extract of Rhus mysorensis, AERM- Aqueous extract of Rhus mysorensis.

 

4. DISCUSSION:

The present study showed a dose dependent efficacy of the plant extracts, as an increase in the concentration of the test extracts caused rapid onset of paralysis followed by death.  It was reported that several bioactive compounds such as flavonoids, alkaloids, tannins, terpenoids etc. are responsible for the anthelminthic activity of a plant extract^10-12. Tannins may interfere with energy generation of worms by uncoupling oxidative phosphorylation or they binds to the free protein of the gastrointestinal tract of the worms¹³, or glycoprotein on the cuticle of the parasite¹⁴, and lead to death. Thus the presence of tannins, triterpenoid and flavonoids might have responsible either alone or together for anthelminthic activity.

 

5. CONCLUSION:

From the above discussion it can be said that all the three extracts of leaves of Rhus mysorensis showed potent anthelminthic activity. It can be concluded that active constituents responsible for anthelmintic activity might be present in the leaves extracts. Further investigations are needed to establish the mechanism of action and isolation of active principles responsible for the anthelminthic activity.

 

6. REFERENCES:

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9.             H. K. Sudeep Kumar et al. Evaluation of anthelmintic activity of Pistia stratiotes Linn. Journal of Basic and Clinical Pharmacy. 2010 May; 1(2): 103-105.  

10.          Pessoa LM, Morais SM, Bevilaqua CM, Luciano JH. Anthelmintic activity of Essential oil of Ocimum gratissimum Linn. And eugenol against Haemonchus contortus. Vet. Parasitol.2002; 109:59-63.

11.          Lahlou M. Potential of Origanum compactum as a cercaricide in Morocco. Ann. Trop. Med. Parasitol. 2002; 96:587-593.

12.          Bhadauria P et al. A review on Saraca indica plant. Int Res J Pharm 2012; 3(4):80-84.

13.          Mute V.M. Anthelmintic effect of Tamarind indica linn leaves juice extract on Pheretima posthuma. Int J Pharma Res and Dev 2009; 7:1-6.

14.          Thompson DP, Geary TG. The structure and function of helminth surfaces. In: J.J. Marr, Biochemistry and Molecular Biology of Parasites, 1st Ed. New York: Academic Press; 1995:203-232.

 

 

 

 

 

Received on 20.05.2016       Modified on 18.06.2016

Accepted on 27.06.2016      ©A&V Publications All right reserved