Investigations of Anthelmintic Activity of Ethanolic Leaf Extract of Premna esculenta

Jannatul Ferdous Shaily¹, Syeda Zakia Sultana¹, Pritesh Ranjan Dash^2* and Taslima Akter^3
*Correspondence: Pritesh Ranjan Dash, Department of Pharmacy, University of Science and Technology Chittagong, Chittagong, Bangladesh, Email:

Keywords

P. posthuma • Anthelmintic • Albendazole

Introduction

Antihelminthics, also known as anthelmintics, are medicines that kill or stupefy parasitic worms (helminths) so they may be eliminated from the body [1]. According to the WHO, an astounding two billion individuals are infected with parasitic worms. Also affecting cattle and crops are parasitic worms that have an economic impact by altering food production. Despite the high frequency of parasitic illnesses, anthelmintic medication research is lacking. According to WHO, only a select few medications are used routinely utilized to treat certain parasite disorders. Traditional medicine claims that several natural herbs are effective at getting rid of worms [2].

Tribal people have historically treated a variety of inflammatory disorders with P. esculenta (Family: Verbenaceae). A shrub with short limbs and branching is known as P. esculenta Roxb. Family Verbenaceae) and is found in the forest areas of Bangladesh and the Chittagong and Chittagong Hill Tracts of India [3]. The herb often referred to as "Lelompata," has long been used by tribal people in Bangladesh to cure Jock worm and appetite infection, frenzy, hepatic, yellow fever, fluor albus, tumor, swelling, serpent bite, abdomen ailments, and kidney stones. Consequently, for the treatment of bacterial and fungal infections, arthritis, and other conditions, the plant’s leaves are applied directly to the area. That is afflicted in traditional medicine. Roots are frequently combined with other plants to treat gout, edema, and jaundice. The leaves are one of the ingredients of a treatment used to cure jaundice in Khagrachari, Bangladesh. For jaundice patients in the Chittagong Hill Tracts, leaves boiled with a Nappi fermented paste made from different marine fish species are a crucial part of their diet [4].

Materials and Methods

The experiment used two substances: pyrantel pamoate and ordinary saline water. In each group, different dosages of ethanol extract from P. esculenta leaves were investigated. The control was regular saline water. The reference medication for the comparison investigation in the in vitro anthelmintic activity was pyrantel pamoate obtained from Beximco Pharmaceuticals Ltd.

Earthworms

In vitro, testing of the anthelmintic action was done using adult earthworms (P. posthuman). A collection of earthworms was made at the IUT in the Gazipur area and were identified at Primeasia University in Bangladesh. The length and breadth of the earthworms were 2-4 cm and 0.2-0.3 cm, respectively.

Collection and identification of plant

The district of Gazipur is where P. esculenta was collected. The accession number for this plant was DACB78766, and it was collected and identified at the Bangladesh National Herbarium in Mirpur, Dhaka. The plant component was properly cleaned with distilled water, and then it was divided into little pieces and dried in the shade.

Preparation of fresh leaves extract

Weighting 75 g of the collected leaves, 150 ml of distilled water was added to make a liquid. The mixture was then centrifuged at a speed of 150 rpm. The supernatant was filtered via sterile filter paper into a conical flask since the research extract 1 ml of filtrate is expected to contain 0.5g or 500 mg/ml.

Phytochemical screening

Using established phytochemical techniques, the freshly obtained crude extract was qualitatively examined for the presence of several phytochemical elements such as alkaloids, tannins, reducing sugar, flavonoids, steroids, terpenoids, and saponins [5,6].

Anthelmintic activity

According to the procedure outlined by Ajaiyeoba EO, et al. the anthelmintic test was conducted [7]. Based on its morphological and physiological similarity to intestinal roundworm a parasite in people, the adult Bangladeshi earthworm (P. posthuman) was used in the experiment in vitro to test for anthelmintic activity. Six earthworms (P. posthuman) of roughly comparable size were introduced in every beaker with 50ml of the aforementioned tests for the extract solutions. The test samples of the extract were made at concentrations of 25, 50, and 100 mg/ ml in normal saline water. Pyrantel pamoate (10mg/ml) served as the control and ordinary saline water as the reference standard. Before beginning the tests, all of the test solutions and the reference medication solution were newly made. When there was no movement visible other than when the worms were violently agitated, the duration of paralysis was recorded [8]. After determining that worms when violently shaken or submerged in warm water, remained still, the time of worm death was recorded (50℃)

Results

P. esculenta's ethanolic leaves extract underwent phytochemical screening, which identified alkaloids, tannins, reducing sugar, flavonoids, steroids, terpenoids, and saponins.

Anthelmintic activity

The ethanolic leaf extract of P. esculenta showed potent anthelmintic action that was equivalent to the prescribed medication Pyrantel pamoate (10 mg/ ml). Results are displayed in Table 2. According to the extract, the most potent anthelmintic activity at the dose of 100 mg/ml at which paralysis and death of earthworms occurred within about 40 and 55 minutes respectively whereas the duration before earthworms get paralyzed and die by pamoate was about 30 and 60 minutes respectively. The results of phytochemical screening and in vitro anthelmintic activity of P. esculent of the fresh leaves extract are shown in (Table 1) and (Table 2) respectively.

Discussion

It is well known that parasitic worm infections of the digestive system in both people and animals have negative health outcomes and decreased resistance to other illnesses as a result. Various chemicals were examined using worm species, including earthworms, Ascaris, nippostrongylus, and tetrakis, to evaluate compounds with anthelmintic action. Earthworms have been utilized often from among all of these species for the first in vitro testing of anthelmintic compounds because of their accessibility and similarity to intestinal "worms" in terms of how they respond to anthelmintics. It has been shown that any anthelmintics that are harmful to earthworms can be considered anthelmintics [9]. Earthworms can move through ciliary movement. A mucilaginous layer made of intricate polysaccharides makes up the earthworm's outer layer. The earthworm may travel freely since this layer is slimy. Any injury to the mucopolysaccharide membrane will expose the outer layer, which inhibits its motion and may result in paralysis. By harming the mucopolysaccharide layer, this approach may result in the death of the worm. In that one literature review phytochemical analysis, it was found that the plant, P. esculenta contains phenolic chemicals which are caused by tannins attaching to free proteins on the cuticle of the parasite in the gastrointestinal system of the host animal and may result in the parasite's death [10,11].

Conclusion

P. esculenta extract shows in vitro anthelmintic activity, according to the current investigation results. After compound isolation, this bark's medicinal efficacy and mechanism of action may be further assessed.

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