In Vitro Anti-helminthic Activity of hydroalcoholic extract of Martynia annua L. and Pentanema indicum

 

Gayathiri. N. M*, Sudhakar P, Manimekalai P, Sabarinath C

Assist Professor, Department of Pharmacology, Swamy Vivekanandha College of Pharmacy,

Tiruchengode 637205.

 

ABSTRACT:

Objective: Modern anti- helmintics has limited the effective control of the parasites due to number of side effects and development of resistsnt in helminthes.This renewed the interest in screening of medicinal plants for their anti-helmintic activity and the present study was design to invastigate the  In vitro  anthelmintic activity of hydroalcoholic extract of Martynia annua L. and Pentanema indicum. Methods: Nine groups each containing three adult Indian earthworms(Phentanema posthuma) of approximately same size were received into 20ml of desired formulation. I group was treated with normal saline and II group was treated with albendazole (20mg/ml) in 1% w/v of CMC as standard and III, Iv and V group was concentrations of hydroalcoholic extracts of Martynia annua L. 10, 20, 50mg/ml and VI,VII and VIII group was  concentrations of hydroalcoholic extracts of Pentanema indicum 10,20,50mg/ml in 1% w/v of CMC. Last ninth group was treated with combination of  Martynia annua L. and Pentanema indicum (50mg/ml and 50mg/ml). Result: Both the extracts show a dose dependent reduction in paralysis and death time of the worm, combination of   Martynia annua L. and Pentanema indicum 1:1 ratio extrcat was found to be more effcetive especially at 50mg/ml and 50mg/ml. Conclusion: The  anti-helmintic activity of extracts was comparable to standard Albentazole , and could be effective against parasitic helminths of humans and animals.The presence of alkoloid, flavonoids, tannins and phenol in the extracts may be contributory to its anti-helmintic activity.

 

 

INTRODUCTION:

Helminthiasis or worm infestation,  is one of the most prevalent disease.^[1] According to the  World health organization, the infectious diseases are the main cause of death and the key agents of the afflicting worldwide^[2]. Helminths infections are among the most widespread infections in humans especially in poverty stricken and developing countries with warm moist environments and poor sanitary conditions^[3] Inspite of the development of anthelmintic resistance in the parasites of higher economical significance, chemotherapy is still used widely for the purpose of controlling the helminthes.^[4]

 

Most wide spread chronic infections are  caused by various species of parasitic worms^[5]. The majority of infections due to helminths are generally restricted to tropical regions and under nourishment, anemia, eosinophilia and pneumonia. Its worldwide prevalance lies between 500 millions to one billion annually approximately^[6].

 

The gastro-intestinal helminthes becomes resistant to currently available anthelmintic drugs so the treatment of this disease is a challenge⁷. Even the most commoly used drugs  piperazine salt have been  shown  to have side effects like nausea, intestinal distrubance and giddiness and the high cost of modern anthelmintics has limited the effective control of these parasites.^[8] This leads to renewed interest in screening of medicinal plants for their gastrointestinal nematodes.^[9]

 

 

The aim of the present study was design to investigate the anti-helmintic activity of plant  Martynia annua L. and Pentanema indicum belongs to family Martyniaceae and Asteraceae exhibit the various therapeutic activities are reported. Martynia annua L. reported activities are analgesic and antipyretic activity, wound healing activity, antibacterial activity, anthelmintic activity of roots, anticonvulsant activity in albino rats, antioxidant and antifertility activity of leaves, antidiabetic activity of flower and CNS depressant activity performed of root.^[10,11]

 

MATERIALS AND METHODS:

Plant collection and authentication:

Martynia annua and Pentanema indicum were collected from the local areas of around the region of Chitharampoondi, erode. The collected plant were shade dried in the laboratory for 7 days. After complete drying the plant were coarsely powdered in a grinding mill and stored at room temperature in a closed air tight container for further use.

 

Plant authentification was obtained from the Botany department, Vivekanandha institute of college of Arts and Science, Tiruchengode.

 

Worm collection:

Adult Indian earthworm Phentanema posthuma were used to evaluate anthelmintic activity in vitro.Earth worms were collected from moist soil and washed with normal saline to remove all fecal matters.

 

Preparation of the extract (maceration method):

The powdered plant was repeatedly extracted using continuous percolation in a maceration method with 250ml of ethanol solvent based on the polarity. The extracts were evaporated at room temperature and preserved properly before subjecting to anthelmintic activity.

 

Preparation of the test sample:

The test sample was prepared by dissolving 10mg, 20mg, 50mg of hydroalcoholic plant extract residue of the  of the Martynia annua L. and Pentanema indicum in 1%w/v of CMC and the standard extract (Albentazole) was prepared by dissolving 20mg of drug in CMC.^[12,13]

 

Evaluation of anthelmintic activity:

The hydroalcoholic extract Martynia annua L. and Pentanema indicum was screened for anthelminitc activity by using adult Indian earthworms (Pheretima posthuma). Pheritima posthuma was selected in view of its anatomical and physiological resemblance with the intestinal round worm parasite of human beings. Different concentration extracts and the standard drug solution were poures in different petridishes. Nine groups each containing  2-3 earth worms of approximately equal size (5-6 cm) was received into 20ml of desired formulation.^[14] The first group serves as control receives Normal saline (0.9% NaCl),  the second group serves as standard Albendazole 20mg/ml in 1% w/v of CMC and the Hydroalcoholic extract of Martynia annua L. and Pentanema indicum 10mg/ml, 20mg/ml, 50mg/ml and combination of MA & PI (50mg/ml and 50mg/ml) of  Martynia annua L. and Pentanema indicum plant extract in 1% w/v of CMC. Observations were made for the time taken for paralysis and death of individual earthworm. Paralysis was said to occur when the worms do not revive even in normal saline. The time death of worm was recorded after ascertaining that worms neither moved when shaken not when given external stimuli.^[15]

 

RESULT AND DISCUSSION:

The predominant effect of albendazole on the worm is to cause a flaccid paralysis that result in expulsion of the worm by paralysis. Albentazole by increasing chloride ion conductance of worm muscle membrane produces hyperpolarisation and reduced excitability that leads to muscle relaxation and paralysis.^[16]

 

From the result, it is observed that hydroalcoholic extract of Martynia annua L. and Pentanema indicum excellent anthelminitic activity at all the concentartion. The hydroalcoholic extract showed more significant effect on paralyzing the worms, in terms of paralysis time and death time, at compared with standard. The earthworms selected for the anthelminitic activity were most sensitive to the combination of  hydroalcoholic extracts of the Martynia annua L. and Pentanema indicum (50mg/ml and 50mg/ml). The table revealed dose dependent paralysis ranging from loss of motility to loss of response to external stimuli, which eventually progressed to death. In case of the hydroalcoholic extracts of the Martynia annua L.10, 20 and 50mg/ml concentrations paralysis was observed respectively at 53.33±4.41, 28.00±6.25 and 25.00±2.89 min and death at 107.33±3.71, 70.00±2.89 and 41.00±2.08 min. The hydroalcoholic extracts of  Pentanema indicum 10, 20 and 50mg/ml concentrations paralysis was observed respectively at 15.00±1.15, 11.33±1.45 and 7.33±1.45min and death at 34.00±2.89, 20.00±3.46 and 14.00±2.31min. The combination  hydroalcoholic extracts of the Martynia annua L. and Pentanema indicum (50mg/ml and 50mg/ml) paralysis was observed respectively at 6.00±1.15 min and death at 12.00±1.15min. The standrad drug Albentazole (20mg/ml) showed paralysis at 20.67±1.76min and death occured after 32±3.21min. The earthworms were more sensitive to the extracts of combination of   Martynia annua L. and Pentanema indicum (50mg/ml and 50mg/ml) concentration as compared to the reference standard drug, Albentazole (20mg/ml). The results were compared with the standard drug, Albentazole and it was found that both extracts were more effcetive than the selected standard drug.

 

The curative properties of medicinal plants are perhaps due to the presence of various secondary metabolites such as alkaloids, terpenoids,phenols and tannins etc.^[17] From above the results, it is concluded that the extracts of the plant have potent anthelmintic activity when compared with the conventionally used drugs and is equipotent to standard anthelmintic drug. Further results, using in vivo models are required to carry out and establish the effectiveness and pharmacological rationale for the use of the plant as an anthalmintic drug.

 

Table 1: Effect of  hydroalcoholic extract of Martynia annua L. and Pentanema indicum on Indian earthworms(Pheretima posthuma)

Treatment Group	Concentration (mg/ml)	Time taken for paralysis (in min)	Time taken for death (in min)
Control (normal saline)	-	-	-
Albendazole	20	20.67 ± 1.76	32.00 ± 3.21
HAEMA	10	53.33 ± 4.41	107.33 ± 3.71
	20	28.00 ± 6.25	70.00 ± 2.89
	50	25.00 ± 2.89	41.00 ± 2.08
HAEPI	10	15.00 ± 1.15	34.00 ± 2.89
	20	11.33  ± 1.45*	20.00 ± 3.46**
	50	7.33 ± 1.45**	14.00 ± 2.31***
HAEMA + HAEPI	50 + 50 (1:1)	6.00 ± 1.15**	12.00 ± 1.15***

Values are expressed as Mean ± SEM, n=3, Values were found out One way ANOVA followed   by paried t-test. Symbol represents statistical significance: * P<0.05 ** P<0.01 *** P<0.001

 

Figure 1: Anthelmintic activity of hydroalcoholic extracts of Martynia annua L. and  Pentanema indicum

 

Figure 2: Anthelmintic activity of different concentration of hydroalcoholic extracts of the Martynia annua L. and Pentanema indicum

 

 

 

CONCLUSION:

The present study justifies the folkloric claims of the potential anthelmintic activity of hydroalcoholic extracts of the Martynia annua L. and Pentanema indicum plant parts. The plant may have different mode of actions against the selected helminthes. The possible mechanism of the anthelmintic activity of hydroalcoholic extracts of the Martynia annua L. and Pentanema indicum cannot be explained on the basis of our present results.The plant may be further explored for its phytochemical profile to recognize the active constituent and standardization of dose and toxicity studies for drug development accountable for anthelmintic activity. Further study is required with the selected plant for the development of novel standardized anthelmintic herbal formulations.

 

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Accepted on 24.04.2019           © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(8):3847-3850.