Assessment of anthelmintic potential of Averrhoa bilimbi, Clerodendrum viscosum and Drynaria quercifolia: as an alternative source for anthelmintics

Md. Ramjan Ali¹*, Marjan Hossain¹, Jannatul Ferdous Runa¹, Md. Hasanuzzaman^{1, 2}

¹Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali- 3814, Bangladesh

²College of Medicine, INJE University, Busan, Republic of Korea.

ABSTRACT:

Three medicinal plants were presented in this study to investigate the anthelmintic activity using earthworms (Pheretima posthuma).Various concentrations (10 mg/ml, 20 mg/ml, 30 mg/ml, 40mg/ml and 50 mg/ml) of three methanolic extracts were tested and results were expressed in terms of time of paralysis and time of death of worms. Albendazole (15 mg/ml) was taken as reference standard and water as control. Paralysis time can be determined by vigorous shaking when no movement was observed. When whitish substances were secreted from the body that one was termed as death time. Dose dependent activity was observed in all extracts but Averrhoa bilimbi showed more activity to Clerodendrum viscosum and Drynaria quercifolia. This is also observed that three plant extracts exhibited more potency than standard.

KEYWORDS: Pheretima posthuma, Albendazole, Dose dependent activity, anthelmintic activity.

INTRODUCTION:

Anthelmintic are drugs that expel parasitic worms (helminths) from the body, by either stunning or killing them. The WHO estimates that an over whelming two billion people having parasitic worm infections. Parasitic worm also infect livestock and crops affecting food production with a subsequent economic impact. In spite of this prevalence of parasitic infections, the research on anthelmintic drug is poor. As per WHO, only few drugs are commonly used in the management of these parasite infections ¹. Parasitic helminthes affects animals and men, causing significant hardship and stunned growth. Most diseases initiated by helminthes are of a chronic; debilitating nature they probably cause more morbidity and greater economic and social deprivation among humans and animals than any single group of parasites ². The major control strategy implemented against helminthes parasite is the use of anthelminths.

Averrhoa bilimbi Linn. belongs to the family Oxalidiaceae having some local name include belembu, belemburi; In English, this is known as- bilimbi, cucumber tree, tree sorrel etc. This possesses short trunk soon dividing into a number of upright branches; attractive, long-lived tropical tree, reaches 16 to 33 ft. (5-10 m) in height. Probably, A. bilimbi is innate of Moluccas in Indonesia. This plant is also obtainable in Brazil, Cuba, Philip-pines, Sri Lanka, Bangladesh, Myanmar (Burma) and Malaysia.³A. bilimbi is medicinally used as a folk therapy for many diseases.

It is generally used as antibacterial, antiscorbutic, astringent; post–partum protective medicine. It is also used for the controlling of fever, mumps, pimples, inflammation of the rectum and diabetes, itches, boils, rheumatism, syphilis, bilious colic, whooping cough, hypertension, stomach ache, aphthous ulcer and as a cooling drink⁴

Clerodendrum viscosum Linn. belongs to family Verbanaceae, frequently known as Bhat in Hindi, Ghentu in Bengali is a terrestrial shrub having square, blackish stem and simple, opposite, decussate, petiolate, exstipulate, coriacious, hairy leaves with a disagreeable odour⁵. The shrub is near 2-4 feet in height. Various parts of the plant are used by tribes for the management of colic, scorpion sting and snake bite, tumors and certain skin diseases. The leaves are somewhat bitter, cure inflammation, skin diseases and good in small pox ⁶.

Drynaria quercifolia J. Smith belongs to family Polypodiaceae, nearby known as Gurar, is a parasitic fern^{7, 8}that is generally distributed in Bangladesh, India and Thailand. The rhizomes of the plant have antibacterial properties and are used traditionally for controlling of cough, tuberculosis and typhoid fever. Asian Centre for Biodiversity identified in their Checklist of Medicinal Plant in Southeast Asia that rhizome decoction or drink of D. quercifolia rhizome uses as antipyretic pre-paration⁹.

MATERIALS AND METHODS:

Collection and identification

Different parts of Averrhoa bilimbi (fruits), Clerodendrum viscosum (leaves), Drynaria quercifolia (leaves) were collected from Comilla, Dhaka and Feni respectively in the month of July 2012 and voucher specimens for each of the collections (DACB 37752, 35979 and 37654 respectively) have been deposited in Bangladesh National Herbarium (BNH) for future references.

Preparation and extraction plant materials

Cold maceration technique was used for extraction. Different parts of plants (500 g) were soaked in 2500 ml of methanol for about 10 days at room temperature with occasional stirring. After 10 days the solutions were filtered using filter cloth and Whatman’s filter paper. The filtrates (methanol extracts) obtained were evaporated under ceiling fan and in a water- bath below 40⁰C until dried. These rendered concentrated portions. The concentrated portions were designated as crude extracts of methanol.

Anthelmintic activity screening

The anthelmintic assay was carried as per the method of Ajaiyeoba et al.,⁹with minor modifications. Adult earthworms (Pheretima posthuma) were used to study the anthelmintic activity due to its anatomical and physiological resemblance with the intestinal roundworm parasite of human beings. Because of availability of earthworms, they are widely used as effective tools for anthelmintic study^10,11. Earthworms were collected from moist soil in the campus of Noakhali Science and Technology University. Collected earthworms were 3 –5 cm in length and 0.1–0.2 cm in width weighing 0.8–3.04 g. They were thoroughly washed with saline water. Pheretima posthuma was placed in petridish containing different concentrations (10-50 mg/ml) of methanolic extract of Averrhoa bilimbi, (fruits) Clerodendrum viscosum (leaves) and Drynaria quercifolia (leaves). Each petridish was placed with 5 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 (15 mg/ml) treated samples.

Table-1: Anthelmintic activity of A. bilimbi, C. viscosum and D. quercifolia

Groups	Concentration (mg/ml)	Time taken for paralysis (P) in min. (Mean and SEM	Time taken for death (D) in min. (Mean and SEM)
Control (Water Only)	-	-	-
Averrhoa bilimbi	10	22.67 ± 0.667	25.33 ± 0.33
	20	16.33 ± 0.33	19.00 ± 0.00
	30	10.33 ± 0.88	13.33 ± 0.33
	40	5.33 ± 0.33	7.33 ± 0.33
	50	2.00 ± 0.00	3.33 ± 3.33
Clerodendrum viscosum	10	82.00 ± 2.00	129.33 ± 1.15
	20	48.667 ± 0.57	115 ± 1.00
	30	45.33 ± 0.57	95.66 ± 1.15
	40	27.667 ± 0.57	81 ± 1.00
	50	18.667 ± 1.15	54.33 ± 2.08
Drynaria quercifolia	10	23.67 ± 1.53	127.67 ± 2.52
	20	14.33 ± 2.64	124.67 ± 6.43
	30	11.00 ± 2.00	96.00 ± 1.00
	40	9.33 ± 3.60	66.33 ± 1.53
	50	6.00 ± 1.73	53.67 ± 4.16
Albendazole	15	35.33 ± 0.57	71.33 ± 1.15

RESULT:

The anthelmintic activity of entire plant extracts of Averrhoa bilimbi (fruits), Clerodendrum viscosum (leaves) and Drynaria quercifolia (leaves) were carried out on earth warm. Different concentrations of three methanolic extracts of different plants were used for the studies. The time taken for paralysis and death of earthworms were recorded in Table–1. The earthworm selected for the anthelmintic activity was most sensitive to the methanol extract of A. bilimbi as can be seen in Fig. 1. The table revealed dose-dependent paralysis ranging from loss of mortality to loss of response to external stimuli, which eventually progressed to death. A. bilimbi was taken 2-22.67 min. to bring paralysis and 3.33-25.33 to bring death of worms. C. viscosum and D. quercifolia also exhibited dose-dependent anthelmintic activity that causes paralysis at 18.66- 82 min. and 6-23.67 min respectively while death at 81-129.33 min. and 53.67- 127.67 min respectively. Albendazole (15 mg/ml) was used as reference standard (paralysis time at 35.33 min. and death time at 71.33 min.). Fig. 1, 2, and 3 represents the anthelmintic activity of A. bilimbi, C. viscosum and D. quercifolia.

DISCUSSION:

The World Health Organization estimates that a amazing two billon people having parasitic worm infections¹². Helminthiasis is still one among the most major human and animal diseases¹³. During the past few decades, despite many advances made in understanding the mode of transmission and treatment of these parasites, there are still no efficient products to control certain helminthes and several the indiscriminate use of some drugs are generated several cases of resistance. As an important component of complementary and alternative medicine, traditional Ayurvedic medicinal plants may be useful to finding and development of new chemical substance for helminthes control which are generally considered to be very important sources of bioactive substances¹⁴. Anthelmintic that are obtained from the natural resources may play an important role in the management of worm infection with less side effects ¹⁵.

From the observations made, different concentration of thee plant extracts produced paralytic and death effect for all worms. Most worm expellers like Albendazole cause paralysis of the worms so that they are expelled in the feces. The methanolic extracts of the three plants not only demonstrated this property but also killed the worms. Methanolic extracts of three plants showed higher anthelmintic activity than standard in a dose dependent manner with highest efficacy at maximum concentration. Therefore, the wormicidal activity of the methanol extracts as described herein against earthworms suggests that these could be effective against parasitic infections of humans.

CONCLUSION:

The wormicidal activity of the methanol extracts suggests that these are effective against parasitic infections of humans. Further, in future it is necessary to identify and isolate the possible active compounds responsible for the anthelmintic activity and study their pharmacological actions.

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Received on 31.03.2013

Modified on 14.04.2013

Accepted on 30.04.2013

Research Journal of Pharmacognosy and Phytochemistry. 5(4): July- August 2013, 178-181