INTRODUCTION:

Punica granatum Linn. (Pomegranate) is a plant of Punicaceae family locally known as Anar. P. granatum is a fruit of great antiquity and is known to have been cultivated in the Middle East more than 5,000 years ago. The plant is found all over India. The fruit of this plant is used as food and as a diet in convalescence, acidosis, dysentery, microbial infections, diarrhoea, helminthiasis, haemorrhage, and respiratory pathologies¹. P. granatum revealed various chemical constituents like ellagic acid, ellagitannins, punicic acid, flavonoids, anthocyanidins, anthocyanins, and estrogenic flavonoids and flavones ² Flavonoid-rich polyphenol fractions from the P. granatum fruit exert antiproliferative,.

The potential therapeutic properties of P. granatum are wide-ranging and include treatment and prevention for cancer³ Cardiovascular disease ⁴Diabetes⁵. Antioxidant⁶ hepatotoxicity⁷_. Pomegranate fruit rind possess excellent antibacterial and anti-diarrhoeal properties⁸Other potential applications include infant brain ischemia, Alzheimer’s disease ⁹ male infertility, arthritis ^10, dermal wounds¹¹ and obesity^12. All of the above medicinal uses either from fruit, seed, or leaves of P. granatum prompted us to investigate the anthelmintic activity of the P. granatum fruit peel extract.

MATERIAL AND METHOD:

Plant material:

The P. granatum Peel materials were collected from local area of Hyderabad and authenticated by Dr. K.Madhava Cheety, Associated Professor, Department of Botany, Sri Venkataeshwara University, (PCOG.H-221).

The plant material was washed under running tap water and dried in shade for 3 weeks. Dried leaves were coarsely powdered, and passed through sieve of mesh size no.22. P. granatum Peel 400 g of coarse powder was extracted with hydro:alcohol (30:70) in Soxhlet apparatus at a temperature not exceeding 60°C. The extract was concentrated under reduced pressure in rotary evaporator to yield a crude semi-solid mass.

Phytochemical evaluation:

Extracts of Puncia granatum were subjected to various chemical tests to detect the phytoconstituents present¹³

Experimental worms:

All the experiments were carried out in Indian adult earthworms (Pheretima posthuma) due to its anatomical resemblance with the intestinal roundworm parasites of human beings. They were collected from moist soil and washed with water to remove all fecal matters.

Preparation of Extracts:

The suspension of Hydro alcoholic Punica granatum Peel extract of different concentrations 100,150,200 mg/ml were prepared by using normal saline and 0.5% w/v of CMC as a suspending agent and final volume was made up to 10 ml for respective concentration. Albendazole was used as standard. Groups of approximately equal size worms consisting of three earthworms individually in each group were released into in each 10 ml of desired concentration of drug and extracts in the petridish.

Experimental Design:

The anthelmintic activity was performed according to the method. On Indian adult earth worm Pheretima posthuma. Each petridish was placed with 3 worms and observed for paralysis or death. Mean time for paralysis was noted when no movement of any sort could be observed, except when the worm was shaken vigorously; the time death of worm (min) was recorded after ascertaining that worms neither moved when shaken nor when given external stimuli. The test results were compared with Reference compound Albendazole (20 mg/ml) treated samples^14.

Group 1: Normal Control Group using normal saline 0.5% w/v of CMC as a suspending agent

Group 2: Standard Group Albendazole (20 mg/ml) using normal saline 0.5% w/v of CMC as a suspending agent

Group 3: 100 mg/kg, Extract using normal saline 0.5% w/v of CMC as a suspending agent

Group 4: 150 mg/kg, Extract using normal saline 0.5% w/v of CMC as a suspending agent

Group 5: 200 mg/kg Extract using normal saline 0.5% w/v of CMC as a suspending agent

RESULTS AND DISCUSSION:

Preliminary phytochemical screening of Hydro alcoholic Punica granatum Peel extract revealed the presence of steroids, flavonoids, saponin, terpenoids, vitamins, alkaloids and tannins. The Hydro alcoholic Punica granatum Peel extract produced a significant anthelmintic activity on Pheretima posthuma worms in a dose dependent manner as shown in Table 1 and Figures (1-5). The peak anthelmintic activity exhibited by Punica granatum Peel extract highest concentration (150,200 mg/ml) which takes 48.71 ± 1.22 for paralysis and 34.90 ± 5.35 minute for death of the worms, followed by extract which includes 74.90 ± 7.42 minute and 62.90 ± 6.85 minute for death of the worms. Potency of the extract was inversely proportional to the time for paralysis (vermifuge) and death (vermicidal) of the worms.

Table 1: Anthelmintic potency of Hydro alcoholic Punica granatum Peel extract

Extract	Concentration (mg/ml)	*Pheretima* *posthuma*
Extract	Concentration (mg/ml)	Paralysis (P)	Death (D)
		Paralysis (P)	Death (D)
Control (Normal saline 0.5% CMC)	-	-	-
Standard (Albendazole)	20 mg/ml	26.71 ± 1.86	38.90 ± 4.85
Punica granatum Peel extract	100 mg/ml	51.90 ± 6.85	90.10 ± 6.32
Punica granatum Peel extract	150 mg/ml	48.71 ±1.22**	74.90 ± 7.42*
Punica granatum Peel extract	200 mg/ml	34.90 ±5.35**	62.90 ±6.85***

Differences were considered significant whenever the P value are reported as mean ± SEM. ***p<0.001, **p<0.01 and *p<0.05.

Figure 1- showing control Group

Figure 2- showing 100mg P. granatum Extract

Figure 3- showing 150mg P. granatum Extract

Figure 4 - showing 200mg P. granatum Extract

Figure 5- showing Albendazole (20 mg/ml)

CONCLUSION:

The Punica granatum Peel extract has showed significant anthelmintic activity at all the tested doses when compared to control as vermifuge and vermicidal while highest activity exhibited by the higher conc. (200 mg/ml) which assures the ethno-medicinal claim Hence, we can think about this herb as alternate source of anthelmintic drugs and also can generate new active lead for suitable anthelmintic drug.

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Received on 30.01.2018 Modified on 05.03.2018

Res. J. Pharmacology and Pharmacodynamics.2018; 10(2): 66-68.

DOI: 10.5958/2321-5836.2018.00011.3