Anti Helminthis Activity of Alangium Salviifolium Against Pheretima Posthumous
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Int.J.Curr.Microbiol.App.Sci (2018) 7(11): 2720-2724 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 7 Number 11 (2018) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2018.711.311 Anti Helminthis Activity of Alangium salviifolium against Pheretima posthumous K.P. Kochuthressia* Department of Botany, Carmel College, Mala (P.O), Thrissur – 680732, Kerala, India *Corresponding author ABSTRACT The present study was intended to delineate the Anti Helminthis activity of Ankolam leaf K e yw or ds extract. The crude extract was obtained by the employment of Soxhlet extraction method. The extract was used to screen the antagonistic activity in vitro by taking Pheretima Alangium salviifolium, posthumous as the study subject. B y the employment of molecular sequencing the identity Anti helminthis activity, Pheretima posthumous of plant was confirmed as Alangium salviifolium. The results obtained proved the capability of extract to down regulate the biological activities of subject. Mortality was Article Info induced by varying concentrations (100-1000 ppm/ml). Interestingly the antagonistic Accepted: activity was in direct proportional with the extract administrated. The analysis of phyto- 22 October 2018 constituents revealed that the extract is composed of polyphenols, Saponins, Flavonoids Available Online: etc. Therefore, it is assumed that the biological activities are complimented by the phyto- 10 November 2018 components. The results obtained from the current studies can be further exploited in the field pharmaceuticals. Introduction (Kumarasingha et al., 2016). Similarly, Helminthis infection causes huge revenue in Despite of some biological advantages in the agro farming. There have been several field of agriculture Helminthis are known for synthetic chemical compounds and its expanded infection and disease spreading nanoparticles proven their anti helminthis abilities. Therefore, it is crucial to improve activity. However, it is also observed that efficient control agents against them (Cabardo these compounds are mostly toxic to the and Portugaliza, 2017). environment. In this juncture it is essential to develop suitable ethnobotanical agents against Trematodes, the causal agents of this threat (Khan et al., 2018). schistosomiasis an important parasites of economic and public health implications. Alangium salvifolium Wang. belongs to the Schistosomiasis affects over 240 million family Alangiaceae and is commonly known people worldwide, with up to 700 million as sage leaved Alangium. It is a well-known individuals living at risk of infection (Cala et traditionally used medicinal plant in India and al., 2012). The disease caused up to 250,000 it is also one of the most versatile medicinal deaths per year in the last decade plant having a wide spectrum of biological 2720 Int.J.Curr.Microbiol.App.Sci (2018) 7(11): 2720-2724 activities like antidiabetic, antiulcer, analgesic, Collection of study subjects anti-inflammatory, antimicrobial, antioxidant, anti-arthritic, diuretic, antifertility, Pheretima posthumous were collected from anthelmintic, antiepileptic and antifungal vermicomposting unit of St. Mary’s College, (Shravya et al., 2017). A. salvifolium is a tall Thrissur, Kerala and washed with normal thorny tree and the genus contains 17 species saline to remove all fecal matter which was of small trees, shrubs and lianas. Alangium brought to Laboratory. The identity of name has been derived from the Malayalam collected organisms was confirmed by the word Alangi. Almost all the parts (root, bark, help of Dr. Dalie Domnic, Head of the leaves, seeds and fruits) are known to have department, Department of Zoology, St. important therapeutic uses and are extensively Mary’s College, Thrissur, Kerala. Study used for different purposes in the indigenous subjects were acclimatized in aquaria for a herbal medicines (Saraswathy et al., 2010). minimum period of four days in holding tanks containing aerated, de-chlorinated tap water A. salvifolium is widely distributed in South and washed sand. East Asia, from India to China, Thailand, Philippines, Indonesia and Papua New Screening of anthelmintic activity Guinea. It is also found in Africa, Madagascar, Southern and Eastern Asia, tropical Australia, Test samples of plant extract was prepared at the western Pacific Ocean islands and New the concentrations 100 - 1000 ppm/ml in 10 Caledonia. In India, it is found throughout the ml of distilled water and five worms of Hyderabad forests and Sitamata wildlife approximately equal size (same type), about 3 sanctuary, Rajasthan (Singh Tanwer and g weight were placed in each petridish Vijayvergia, 2014) containing 10 ml of above test solution of extracts. All the test solutions were prepared Materials and Methods freshly before starting the experiments. Observations were made for the time taken for Plant collection, identification and extract paralysis was noted when no movement of any preparation sort could be observed except when the worms were shaken vigorously. Time for death of The leaf of Ankolam were collected from treating worms were recorded after Botanical garden of St. Mary’s College, ascertaining that worms neither moved when Thrissur and the identity of plant was shaken vigorously nor when dipped in warm confirmed by molecular sequencing (Zhang et water. Standard drug Albendazole is taken as al., 2016). The dried leaves were ground to a control for comparison (Jacob, 2018). course powder and 50g of the powdered plant materials is subjected to extraction by using Phytochemical analysis of plant extract Soxhlet apparatus using 95% ethanol as the solvent. Phytochemical screening was carried out to assess the qualitative chemical composition of The filtrate is collected after 72 hours. It is crude ethanolic extracts of Alangium twice filtered using Whatman no.1 filter paper salviifolium. Standard screening tests using and extracts were concentrated by evaporation conventional protocol, procedure, and reagents at room temperature. The dried residue is kept were conducted using standard procedures to in refrigerator for further studies (Vadakkan et identify the constituents as described al., 2018). previously (Belemlilga et al., 2016). 2721 Int.J.Curr.Microbiol.App.Sci (2018) 7(11): 2720-2724 Results and Discussion the identification of plants (Vadlapudi and Kaladhar, 2012). Identification of source plant Effect of plant extract in causing death and By the employment of rbcL gene sequencing paralysis in Pheretima posthumous and phylogenetic tree construction it was evident that the source plant is Alangium Ethanolic extract of Alangium salviifolium salviifolium (Figure 1). possess dose dependent anthelmintic action. At higher concentrations, it produces paralytic The BLAST results suggested that source effect much earlier and the time to death at plant is showing maximum similarity that with higher doses, the plant extract induces 100% Alangium salviifolium ribulose-1,5- mortality (Figure 2). It was observed that time bisphosphate carboxylase/oxygenase large required for inducing paralysis and causing subunit (rbcL) gene, partial cds; chloroplast death is in directly proportional that with the which is submitted under the accession concentration of extract administrated. number JF308648.1 and Alangium kurziiribulose 1,5-bisphosphate carboxylase Time taken for inducing paralysis was only 19 large subunit (rbcL) gene, partial cds; minutes when the concentration was 1000 chloroplast gene which is submitted under the ppm/ml whereas the time of survival was accession number AF384108.1. Molecular prolonged to 67 minutes in lower characterization is used as a successful tool for concentration of antagonist. Fig.1 Phylogenetic tree of Alangium salviifolium source plant Fig.2 Antihelmintic activity of extract in varying concentration 2722 Int.J.Curr.Microbiol.App.Sci (2018) 7(11): 2720-2724 A similar kind of activity was reported in I.P., 2016. Anthelmintic activity of aqueous leaf extract of Annonamuricata L. Saba senegalensis (A.DC.) Pichon (Annonaceae) against Haemonchus contortus (Apocynaceae) extract against adult from sheep (Ferreira et al., 2013). Similarly worms and eggs of Haemonchus plant extracts from Brazilian savanna also contortus. Asian Pac. J. Trop. Biomed. possessed Anthelmintic activity(Oliveira et 6, 945–949. https://doi.org/10.1016/ al., 2017). Generally, Albendazole eliminate j.apjtb.2016.07.015 worm chiefly by inducing flaccid paralysis Cabardo, D.E., Portugaliza, H.P., 2017. interestingly plant extract also showed same Anthelmintic activity of Moringa clinical symptoms therefore we assume that oleifera seed aqueous and ethanolic the mechanism of plant extract is similar to extracts against Haemonchus contortus that of Albendazole (Haque et al., 1993). eggs and third stage larvae. Int. J. Vet. Sci. Med. 5, 30–34. https://doi.org/ Phytochemical analysis of plant extract 10.1016/j.ijvsm.2017.02.001 Cala, A.C., Chagas, A.C.S., Oliveira, M.C.S., Phytochemical screening of ethanolic extract Matos, A.P., Borges, L.M.F., Sousa, of Alangium salviifolium showed the presence L.A.D., Souza, F.A., Oliveira, G.P., of alkaloids, coumarins, terpenoids, 2012. In vitro Anthelmintic effect of cardioglycosides, flavonoids, saponins, Melia azedarach L. and Trichilia quinine and phenols as major chemical claussenii C. against sheep constituents. Among these, poly phenols, gastrointestinal nematodes. Exp. flavonoids are said to have anthelmintic Parasitol.