INTRODUCTION:

Plants have been used for medicinal purposes for as long as history has been recorded. Despite the progress in orthodox medicine, interest in alternative medicine, including herbalism, is on the increase in the West and for 80% of the world herbal medicine is still the only kind to which ordinary persons have ready access. A great variety of plants are used for medicinal treatments. Either the dried plant, or a specific part of it (root, leaves, fruit, flowers, seeds), is formulated into suitable preparations — compressed as tablets or made into pills, used to make infusions (teas), extracts, tinctures, etc., or mixed with excipients to make lotions, ointments, creams.⁽¹⁾

Similarly, many consider that since plants are natural materials they are safer and will produce fewer side-effects than synthetic drugs. There is little substance or reason in either of these claims. For example, comfrey (Symphytum officinale) is considered a safe herb and is used as a demulcent. However, it contains pyrrolizidine alkaloids, which are toxic to the liver and can cause liver cancer. Herbal medicine (or "herbalism") is the study and use of medicinal properties of plants. Studies show that in tropical climates where pathogens are the most abundant, recipes are the most highly spiced. Further, the spices with the most potent antimicrobial activity tend to be selected. In all cultures vegetables are spiced less than meat, presumably because they are more resistant to spoilage.⁽²⁾

Among the more popular remedies used are ginseng, to increase stamina and as a mild sedative; St.-John's-wort, for mild depression; echinacea, to aid the immune system and alleviate colds; kava, to calm anxiety and treat insomnia; saw palmetto, for enlarged prostate; and ginkgo biloba, to improve short-term memory.⁽³⁾

This widespread use has prompted demands that herbal remedies be regulated as drugs to insure quality standards. The U.S. Food and Drug Administration (FDA) can require a clinical trial on any herb that has a health claim on its label, but medical testing, which is geared toward observing a particular active component, is difficult to apply to herbs, which may have many interacting ingredients.⁽⁴⁾

Plants have the ability to synthesize a wide variety of chemical compounds that are used to perform important biological functions, and to defend against attack from predators such as insects, fungi and herbivorous mammals. Many of these phytochemicals have beneficial effects on long-term health when consumed by humans, and can be used to effectively treat human diseases. The World Health Organization (WHO) estimates that 80 percent of the population of some Asian and African countries presently uses herbal medicine for some aspect of primary health care.⁽⁵⁾

Couroupita guianensis is a deciduous tree belonging to the family Lecythidaceae. It is native to South India and Malaysia. Various part of the tree have been reported to contain oils, keto steroids, glycosides, couroupitine, indirubin, isatin and phenolic substances The pulp of the fruits oxidizes bluish-green when exposed to air and is extremely malodorous probably because of sulphur compounds in the fruits.⁽⁶⁾ The fruit contains small seeds in a white, unpleasant smelling edible jelly. The large fruit, which is woody and very spherical, measuring up to 25 centimeters wide, gives the species the common name "cannonball tree". A smaller fruit contains perhaps 65 seeds, while a large one can have 550. One tree can bear 150 fruits. The fruit takes up to a year to mature in most areas, sometimes as long as 18 months. This plant is used for treating mange and other skin conditions. The pulp of the fruit of the cannon ball tree is rubbed on the infected skin of mange dog. It is claimed that when the dog licks its skin, this medicine will also work internally. The flowers are used to cure cold, intestinal gas formation and stomachache.⁽⁷⁾

PLANT PROFILE

Scientific Name:

Couroupita guianensis Aubl.

Common Names:

Cannon ball tree, Sal tree, Ayauma tree

Ethnomedicinal uses:

Ø Leaves of C.guianensis are widely used as an analgesics by the Brazilian rural population

Ø Juice made from the leaves is used to treat skin disorders and the Shamans of South America have even utilized tree parts for curing malaria

Ø Couroupita guianensis infusions or teas obtained from different parts of the tree used traditionally to treat hypertension, tumours, pain and inflammatory processes

Ø Couroupita guianensis flowers are used to cure cold,intestinal gas formation and stomach ache

Ø In orissa,decoction of flowers has been used to boost the immune system to fight number of diseases

Ø The fruit pulp of Couroupita guianensis is used to disinfect wounds

The plant was collected from Bobbili region, Vizianagaram, in the month of December, The leaves were washed thoroughly with sterile distilled water in order to remove any dirt or filthy particles present on the surface and were shade dried at room temperature for about 10 days. The shade dried leaves were pulverized in mixer grinder to form fine powder and passed through mesh size 100.

Three slides are prepared accordingly with different reagents as following and seen under compound microscope

1.T.S.+Glycerin

2.T.S.+Iodine solution+Glycerin

3.T.S+Phloroglucinol+Con.Hcl+Glycerin

The transverse section of the leaf of couroupita guianensis shows the presence of:

· Starch grains

· Anomocytic stomata

· Prism type of calcium oxalate crystals

· Covering trichomes

· Xylem vessels

Preparation of Extracts by successive solvent extraction

The finely powdered leaf drug of couroupita guianensis about 80gm was extracted with chloroform(50-55 c) for 72 hours by continuous hot percolation method using soxhlet apparatus.Then it was evaporated to form a dry mass of chloroform extract.

Aqueous extract is prepared by dissolving 500ml distilled water in 500gm of finely powdered leaf drug of couroupita guianensis and kept for 36hrs.Then it is filtered and evaporated.

EVALUATION OF ANTHELMINTIC ACTIVITY OF LEAF EXTRACTS OF couroupita guianensis:

Chloroform and aqueous extracts from the leaves of Couroupita guianensis were investigated for anthelmintic activity against Pheretima posthuma. Various concentrations (100 and 200 mg/ml) of each extract were tested by bioassay, which involved determination of time of paralysis and time of death of the worms. Albendazole was used as standard reference and distilled water as control. The Anthelmintic assay was carried as per the method followed by Ajaiyeoba et al with minor modifications^[8]. The assay was performed on adult Indian earth worms, Pheretima posthuma due to its anatomical and physiological resemblance with the intestinal round worm parasite of human beings^{[9, 10].} Because of easy availability, earthworms have been used widely for the initial evaluation of anthelmintic compounds in vitro ^{[11, 12]}. The earthworms were collected from moist soil and washed with normal saline to remove all faecal matter and were used for the anthelmintic study. The earth worms of 6-8 cm in length and 0.2-0.3 cm in width were used for all experimental protocol. The earth worms were divided into ten groups containing six earth worms in each group. All the extracts and standard drug solution were freshly prepared in normal saline before starting the experiments. Different extracts and standard drug solutions were poured in different petri plates. All the earth worms were released into 10ml of formulation as follows: Ethanol extract, Aqueous extract, Albendazole in three different concentrations. Observations were made for the time taken to paralysis and death of worms.^(13,14) Time for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Death was concluded when the worms lost their motility when dipped in warm water (50ºC) followed with fading away of their body colors.

RESULTS AND DISCUSSION:

Qualitative analysis of bioactive compounds in different solvent extracts of Couroupita guianensis leaves

Test name	Chloroform extract	Aqueous extract
Mayer’s	++	+
Wagner’s	++	+
Dragendroff’s	++	+
Tannins	+	+
Phlobatannins	++	+
Glycosides	++	++
Sterols	++	+
Resins	++	+++
Phenols	+	+
Anthraquinones	++	+
Carbohydrates	++	++
Cardiac glycosides	-	_
Steroids	+	+
Terpenoids	++	++
Alkaline reagent Test	+	+

Anthelmintic potency of chloroform and aqueous extracts of Couroupita guianensis leaves.

Extract	Concentration (mg/ml)	*Pheretima posthuma*
Extract	Concentration (mg/ml)	Paralysis (P) (sec)	Death (D) (sec)
Control (distilled water)	-	-	-
Standard (Albendazole)	20	140	120
Chloroform extract	100	1800	1500
Chloroform extract	200	1200	1100
Aqueous extract	100	3300	975
Aqueous extract	200	2900	540

DISCUSSION:

Preliminary phytochemical analysis of chloroform extract and aqueous extract showed the presence of Flavonoids, Glycosides, Alkaloids, Steroids and TriTerpenoids whereas the aqueous extract revealed the Tannins, Glycosides and Alkaloids as active phytochemical constituents.⁽¹⁵⁾The data revealed that the chloroform extract showed better anthelmintic activity at a concentration of 200 mg/ml, whereas the aqueous extract also showed paralysis and death at similar concentrations against Indian earth worms. The other test concentrations of both the extracts showed marked degree of anthelmintic activity. The anthelmintic effect of extracts is comparable with that of the effect produced by the standard drug albendazole. Screening and proper evaluation of alternatives that may be both sustainable and environmentally acceptable. The results of this study have shown promising anthelmintic activity suggesting the possible use of extracts in intestinal nematode control.

CONCLUSION:

The results of the present study clearly indicated that the crude chloroform and aqueous extracts of Couroupita guianensis did produce anthelmintic activity against Indian earthworm Pheretima posthuma .The plant possesses significant anthelmintic activity at 200 mg/ml concentration measured by time taken for paralyse / death of the earth worms. The current investigation leads to conclusion that the leaves of Couroupita guianensis have potent anthelmintic activity when compared with the conventionally used drug. The results did not, however, exclude the possibility that doses of the extract with lower anthelmintic activity in this study might be efficacious against other species of helminths. Further studies using in vivo models and to isolate active constituents from extract are required to carry out and established the effectiveness and pharmacological rational for the use of Couroupita guianensis as an anthelmintic drug. There is growing interest in correlating the phytochemical constituents of a medicinal plant with its pharmacological activities.

The present study was carried out with a vision to setup standards that could be beneficial for detecting the authenticity of this vital medicinal plant. Numerical standards reported in this work could be useful for the compilation of a suitable monograph of Couroupita guianensis. As the plant produce secondary metabolites in order to protect themselves from microorganism, herbivores and insects, thus antimicrobial effect is somehow expected from plants namely flavonoids, alkaloids, tannins, saponins and glycosides are producing a better opportunity for testing wide range of microorganism. The results obtained from this work revealed that the plants contained bioactive agents which are connected with anthelmintic properties in plants.

Since Couroupita guianensis is easily available and well tolerated, it can be incorporated into medications for anthelmintic therapy. However, further studies for its incorporation into oral preparations, safety and cost- effectiveness has to be conducted.

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9. Mariana M.G. Pinheiro, Sidnei B.O. Fernandes, Catarina E. Fingolo , Fábio Boylan, Patrícia D. Fernandes .” Anti - inflammatory activity of ethanol extract and fractions from Couroupita guianensis Aublet leaves.”Journal of Ethnopharmacology. Vol. 146:Issue 1,7 march 2013.

10. Sanjay. Prahalad Umachigi, Jayaveera K.N., Ashok kumar C.K., G.S. Kumar.” Antimicrobial, Wound Healing and Antioxidant potential of Couroupita guianensis in rats “. (2007).page no: 269.

11. Patrícia Dias Fernandes, Mariana M.G. Pinheiro , Sidnei B.O. Fernandes , Catarina E. Fingolo , Fábio Boylan.” Antinociceptive activity of fractions from Couroupita guianensis Aubl. Leaves.”Journal of Ethnopharmacology.Vol.127, Issue 2,3 February 2010.

12. Mariappan Premanathan, Srinivasan Radhakrishnan, and Kandasamy Kathiresan. “Antioxidant and anticancer activities of isatin (1H-indole-2,3-dione), isolated from the flowers of Couroupita guianensis Aubl”. Indian J Med Res.2012 November;136(5):822-826.

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Received on 22.06.2016 Modified on 30.06.2016

Res. J. Pharmacology & Pharmacodynamics.2016; 8(3): 118-122.

DOI: 10.5958/2321-5836.2016.00022.7