In Vitro Anthelmintic Activity of Various Herbal Plants Extracts against Pheritima posthuma

 

Priyanka Patil1*, J.K. Patel2, P.S. Kulkarni1, M.U. Patel1,  C.J. Bhavsar3 and Patel J.A.1

 

1P.S.G.V.P Mandal’s College of Pharmacy, Shahada (M.S), India

2Nootan Pharmacy College, Visnagar, Gujarat, India

3H.R.Patel college of Pharmacy, Shirpur

 

 

ABSTRACT:

Aqueous extracts of leaves of Cissus quadrangularis, Eclipta alba, Chrozophora rottleri and flowers of Luffa acutangula were investigated for anthelmintic property against Pheritima posthuma (Indian Earth worm). Different concentrations (25, 50 and 100 mg / ml) of each plants aqueous extracts were studied in a bioassay, which involved the determination of time of paralysis and time of death of the worms. Piperazine citrate (10 mg/ml) was used as a standard reference drug. All the aqueous extracts of the three plants exhibited significantly anthelmintic activity against Pheritima Posthuma, but flowers of Luffa Acutangula demonstrated the best anthelmintic activity in both the parameters.

 

KEYWORDS: Aqueous extracts,  Cissus Quadrangularis, Eclipta alba, Chrozophora rottleri, Luffa acutangula, anthelmintic and Pheritima posthuma

 

1. INTRODUCTION

Diseases caused by helminth parasites in continue to be a major productivity constraint, especially in small ruminants in the tropics and subtropics1. In the developing world, the greatest impact of parasitic diseases is indirect and potential productivity losses1. Infections by gastrointestinal helminth parasite of livestock are among the most common and economically important diseases of grazing livestock2.  Adulteration of anthelmintic has been found to be common practice3. Illiteracy and unfamiliarity with synthetic anthelmintic, resulting in incorrect usage, are also a problem leading to the same consequence.  Moreover, these drugs are relatively expensive.  As a consequence of these problems and difficulties, pastoralists and small holder farmer have continued to use indigenous plants as livestock dewormers4. Considerable research has shown that some plants not only affect the nutrition of animals, but also have antiparasitic effects5. For Example, plants that contain condensed tannins, a class of phenolic secondary metabolite, have these effects. Cissus quadrangularis Linn. also known as Hadjod, Asthisamhari, Asthi-shrankhala, Asthi-samjaka in Indian vernacular, belongs to family Vitaceae.The allied names refer to its effects on skeletal system, most significant being in healing of fractures. Animal studies and human trial showed promising result of Cissus quadrangularis in fracture healing and osteoporosis6.The plant Eclipta alba is an indigenous medicinal plant has folk reputation in rural area of southern India, literature survey reveals presence of steroidal alkaloid, ecliptalbine 4 beta hydroxyverazine. The leaves of Eclipta alba Linn. have been reported for antibacterial, analgesic and anticonvulsant activities7. The Chrozophora rottleri (Suryavarti )is an erect herb with silvery hairs. The leaves are densely hairy on both sides8.


Literature describing the chemical nature is not available.  Luffa acutangula is one of the important plant mentioned in Ayurveda and Unani 9. The flowers of Luffa acutangula have been folk used as an anthelmintic10. The juice of fruit of Luffa acutangula use as emetic, expectorant, purgative, demulcent and diuretic11. The present study is based to identify the potential anthelmintic property along with comparative details of various herbal plants using Pheritima posthuma (Indian Earth worm).

 

2. MATERIAL AND METHODS:

2.1 Plants Collection and Authentication

The leaves of Cissus quadrangularis, Eclipta alba, Chrozophora rottleri and flowers of Luffa acutangula have been collected from the local area of Nandurbar (North Maharashtra, India). All the plants were authenticating by Regional Research Laboratory, Jammu, India.

 

2.2 Plant Extraction:

Collected leaves and flowers were dried and crushed to a coarse powder macerated with water. Extract was dried over anhydrous sodium sulphate and solvent was removed in vacuum at 40°C by using rotary evaporator (Rotavapour Buchii, Switzerland). The aqueous extract was subjected to preliminary phytochemical testing for the presence of different chemical classes of compounds12.

 

2.3 Worms Collection and Authentication:

Indian earthworm Pheritima posthuma (Annelida) were collected from the water logged areas of soils Indian earthworms, identified authenticated in the Department of Zoology, P.S.G.V.P. Mandal’s, Shahada, Maharashtra, India.

 

2.4 Anthelmintic Activity:

The Anthelmintic assay was carried as per the method of Ajaiyeoba et al. with necessary modifications13. The assay was performed on adult Indian earthworm Pheritima posthuma, due to its anatomical and physiological resemblance with the intestinal round worm parasite of human being14-15. Because of easy availability, earth worms have been used widely for initial evaluation of anthelmentic compounds in vitro16. Fifty ml of formulation containing different concentration of crude aqueous extract (25, 50 and 100 mg/ml in distilled water) were prepared and 6 worms of same type were placed in it. 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 were recorded after ascertaining that worms neither moved when shaken vigorously nor when dipped in warm water (500C). Piperazine citrate (10 mg/ml) was used as reference standard while distilled water as control.

 

 

2.5 Statistical Analysis:

The data presented as Mean ± SEM. The activities of all extracts were compared with the control. All the extracts showed significantly activity with higher duration of paralysis and death. Values of P<0.001 were considered statistically significant17.

 

3. RESULTS AND DISCUSSION:

Phytochemical screening of crude aqueous extracts of leaves of Cissus quadrangularis, Eclipta alba, Chrozophora rottleri and flowers of Luffa acutangula revealed the presence of alkaloids, saponins, flavonoids and tannins. As shown in Table 1 and Figures 1-4, aqueous extracts of all the plants exhibited anthelmintic activity in dose dependent manner giving shortest time of paralysis (P) and death (D) with 100 mg/ml concentration, for Pheritima posthuma worms. The aqueous extract of flowers of Luffa acutangula (Figure 3) caused paralysis is 10 min and time of death is 13 min while aqueous extracts of leaves of Cissus quadrangularis (Figure 1), Eclipta alba (Figure 2), Chrozophora rottleri (Figure 4). Revealed paralysis of 16, 17 and 12 min. and time of death 28, 37 and 18 min. respectively against Pheritima posthuma. The reference drug Piperazine citrate showed the paralysis at 22 min. and time of death at 100 mg conc. 49 min. respectively.

 

 

 

Table 1:- Anthelmintic activity of aqueous extracts of various herbal pants on Pheritima posthuma (Indian Earth worm)

Groups

Concentration

(mg/ml)

Time taken for Paralysis (P) and Death (D) of Pheritima posthuma

worms in min (Mean ± SEM)

P

D

Piperazine citrate

10

21.56± 0.34

48.70 ± 0.78

Aqueous extract of Cissus quadrangularis

25

54.6 ± 0.50

64.04 ± 0.57

50

32.6 ± 0.43**

50.72 ± 0.44**

100

16.2 ± 0.37**

27.92 ± 0.19**

Aqueous extract of Eclipta alba

25

62.50 ± 0.67

98.38 ± 0.0.43

50

48.33 ± 0.42*

61.05 ± 0.0.56*

100

17.50 ± 0.43**

37.17 ± 0.0.45**

Aqueous extract of Luffa acutangula

25

61.66 ± 0.50

134.9 ± 0.78

50

28.48 ± 0.19**

50.36 ± 0.52**

100

10.04 ± 0.10***

13.28 ± 0.31***

Aqueous extract of Chrozophoora rotleri

25

82.70± 0.19

150.1± 0.11

50

43.32± 0.33*

56.94± 0.12*

100

11.92± 0.31 ***

18.42± 0.15***

All values are Mean ± SEM; n=6 in each group. Values are significantly different from reference standard (Piperazine citrate) p<0.05; **p<0.01; ***p<0.001

 

 

Piperazine citrate by increasing chloride ion conductance in worm muscle membrane produces hyperpolarisation and reduced excitability that leads to muscle relaxation and flaccid paralysis18. The aqueous extract of flowers of Luffa acutangula not only demonstrated paralysis, but also caused death of worms especially at higher concentration of 100 mg/ml, in shorter time as compared to reference drug Piperazine citrate. Phytochemical analysis of the crude extracts revealed the presence of tannins among the other chemical constituent within them. Tannins were shown to produce anthelmintic activities19. Chemically tannins are polyphenolic compounds20. Some synthetic phenolic anthelmintics e.g. niclosamide, oxyclozanide, bithionol etc., are reported to interfere with energy generation in helminth parasites by uncoupling oxidative phosphorylation 21. It is possible that tannins contained in the aqueous extract of flowers of Luffa acutangula produced similar effects. Another possible anthelmintic effect of tannins is that they can bind to free proteins in the gastrointestinal tracts of host animal22 or glycoprotein on the cuticle of the parasite23 and may cause death.

 

The traditional medicines hold a great promise as source of easily available effective anthelmintic agents to the people, particularly in developing countries, including in India. It is in this context that the people consumed several plants or plant derived preparation to cure helminthic infections24. The origin of many effective drugs has been found in the traditional medicines practices and in view of this it is important to undertake studies pertaining to screening of the folklore medicinal plants for their proclaimed anthelmintic efficacy.

 

4. CONCLUSION:

From the results, it is concluding that the plants Cissus quadrangularis, Eclipta alba, Chrozophora rottleri and Luffa acutangula showed significant anthelmintic activity when compared with the standard anthelmintic drug. Among three plants, aqueous extract of flowers of Luffa acutangula showed best anthelmintic activity as compared with other herbal extracts and standard drug in the study. Further, it would be interesting to isolate the possible phytoconstituents, which may be responsible for the anthelmintic activity and to reveal the mechanism (s) of actions.

 

5. ACKNOWLEDGEMENT:

Authors express their sincere thanks to P. S .G. V. P. M’s College of Pharmacy, Shahada, Dist-Nandurbar, for providing necessary facilities to carry out this research work. Also thankful to Shri Suresh G.  Patil and Dr. Santosh Tayade for their kind support.

 

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Received on 24.02.2010

Accepted on 07.04.2010        

© A&V Publication all right reserved

Research Journal of Pharmacognosy  and Phytochemistry. 2(3): May-June 2010, 234-237