Equisetum diffusum its Phytochemistry, Antimicrobial and Antifungal activity

Equisetum diffusum was analysed for the presence of various phytochemicals and it had been found that the concerned plant is enriched with many phytochemicals with wide range of applications in daily life. In addition the plant was found to good antifungal activity against Aspergillus flavus, Aspergills Niger, Fusarium, Nigrospora oryzae. The concerned plant being anti fungal in nature is one of its best natures as least number of phytochemicals is antifungal in nature. In addition the plant extracts show antimicrobial activity against the concerned bacteria viz, E.coli, Bacillus cereus, Salmonella Typhi, Bacillus pumilus, Bacillus licheniformis, Micrococcous luteus, Streptococcus mutans. The plant extracts show a good range of Rf values where a good number of phytochemicals are isolable with potential medicinal properties.


Introduction
Medicinal plants have been considered and are mentioned from times immoral for treatment of various diseases. It had been known that about 250,000 to 500,000 plant species are under use (Borris, R. P. 1996). But a very little concentration (1 to 10%) of medicinal plants are being consumed as a feeding materials by humans and other animals, but a large portion of these plants are used medicinally ( The genus Equisetum there are 30 species which are rush like, jointed conspicuously, and are perennial. Equisetum is the only living genus of the order Equisetales belonging to class Sphenopsida. Equisetum arvense, possesses tooth like sheath which cover its joints; and from the sheaths of central stem whorls arise, attaining a 60cm in height, but usually less (Clute W.N. 1928, Great Plains Flora Association. 1986. Amny studies showed that the plant is hypoglycemic (Iyer H. 2006, Soleimani S., et.al 2007 and diuretic (Mamedova K.T. et.al., 1996, Jung H.C., et.al., 1999. The plant possesses an anti-inflammatory property and is being used for bathing purpose in Europe, Asia and America and is found to be antiseptic used in Turkey and America (Hoffman D. 1990, Ody P., and Kindersley D. 1993, Mineo S., et.al., 1993. The Equisetum arvense water and ethanolic extracts have the potential to neutralize the free radicles so is being considered to have an antioxidant property (Nagai T., et. Al., 2005). The hydro alcoholic extract have sedative effect (Santos et. al., 2005). As already pointed that, all living horsetails are considered and have been kept in genus Equisetum. But few fossil species are not assignable to the modern genus.

Materials and methods
The plant Equisetum diffusum in fresh form have been collected from a local area bank of the river from the sandy soil at Dehradun. The authentication of the plant was done at BSI, Northern Regional Centre and Dehradun. The plant samples were submitted at B.S.I herbarium with specimen no. 113520. The plant parts wer shade dried at room temperature and then were crushed into powder.
Extraction 50 Gms of plant powder was segregated shade dried, weighed and finally extraction was done by Soxhlet Apparatus. Various solvents with different polarity index (Petroleum ether, Chloroform, Ethanol and Water) were used as per their polarity index. Equisetum diffusum was first extracted by using petroleum ether as a solvent. The respective extract was then filtratered and the obtained filtrate was evaporated to 1/4 th its volume using water bath. The extract mixture after the solvent removal, was made a powder be keeping in an oven for a required time. The remained petroleum ether residue was then subjected to further extraction process using chloroform as solvent in a same way as used for extraction using petroleum ether. Similarly the extraction of the residue after extraction with chloroform was then extracted in a same manner with ethanol. ISSN: 00333077 5600 www.psychologyandeducation.net The residue after extraction with chloroform was then extracted using water as a solvent by decoction technique, in which 500ml of water was added to the residue. The complete mixture was heated using water bath and whole water was made to evaporate, and then more 500 ml of water was further added to the extract, the mixture was then further evaporated in which only 250ml of water was kept in the mixture. The extracted mixture was then filtered, and the obtained filtrate was evaporated completely to 1/4 th of its volume. Finally the extracted filtrate was collected and made to dry by keeping in an oven at 30-50 0 C temperature range. An antimicrobial medicine is a drug that does posses the potential to neutralize and kill the microbes which include fungi, bacteria and viruses. The Antimicrobial medicines either directly kill the microbes so are called microbicidal or may pause the microbial growth so are called microbistatic. The term chemotherapy is used for the treatment of microbial infection

The Microorganisms Source
The antimicrobial assay was performed under the headings of antibacterial and antifungal assays.

Evaluation of the antimicrobial potential of plant extracts:
Five gram positive and three gram negative bacteria viz. E.coli, Bacillus pumilus, Bacillus cereus, Bacillus licheniformis, Pseudomonas aeruginosa, Salmonella typhi, and Streptococcus mutans were taken to evaluate antibacterial potential of the different extracts. All the isolated extracts were analysed for the antimicrobial activity to know whether they are active against the particular bacteria or not. The extracts were dissolved in 30% DMSO, a highly polar, aprotic organic solvents. The food material which is being used as a substrate by microorganisms called culture medium and the growth is called culture.

Evaluation of the antifungal potential of plant extracts:
The extracts were also analysed as antifungal agents against, Aspergillus flavus, Aspergillus Niger, Fusarium and Nigrospora oryza.

Observations and Results
The extraction results of Equisetum diffusum shows the highest percentage yield for aqueous medium providing 13.39% and the Ethanol extract was found 4.99%. The percentage yield of petroleum ether extract was found 2.67% and the lowest yield was found in chloroform extract i.e. 1.28%.         (Vesna M, et. al., 2007). Also the polar solvent extracts like ethanol and water posses more antimicrobial activity as compared to less polar solvent extracts gets verified by the results of (Radulovi, N. et.al, 2006) Analysis of antifungal activity mentioned that the chloroform extracts showed inhibition against all the fungi species in with maximum zone of in 15mm against the fusarium. The Petroleum ether extract was found more effective against all fungi showing max zone of inhibition of 14mm against Nigrospora oryza. The Ethanolic extract also records highest zone of inhibition of 15mm against Aspergillus Niger and Nigrospora oryza and not effective against Fusarium. The Aqueous extract shows minimum zone of inhibition in comparison to all extracts and unveils the growth inhibition of Fusarium. Although the antifungal activity of Equisetum diffusum has been explored yet, but few species of equisetum does posses the antifungal property as per the studies carried out by (Sandhu, NS. et. al., 2010). The antifungal activities of Equisetum diffusum run parallel to the antifungal activity of Equisetum arvense as per the resulted mentioned by (Asgarpanah. J, 2012). As per the thin layer chromatography a good no of Rf values were obtained which corresponds to a particular component. So the study under analysis showed that the extracts are enriched profoundly with wide no of phytochemicals.

Conclusion
The plant in reference shows good antimicrobial and antifungal activity as per the results is taken into consideration. Plants are the ultimate source of all types of chemicals which do have preferential activity against all types of diseases which visit human life from time to time. The concerned plant as per the observation may provide a hope for its use as a wide spread use in treating various diseases. Also further work is required to isolate the bio active compound(s) which may have good antimicrobial properties and have lesser side effect as compared to the market available synthetic drug.