EFFICACY OF AQUEOUS AND ETHANOL EXTRACTS CASUARINA EQUISETIFOLIA FOR POTENTIAL ANTIMICROBIAL ACTIVITY

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Kumar et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 5.210

Volume 4, Issue 07, 1877-1882.

Research Article

ISSN 2278 – 4357

EFFICACY OF AQUEOUS AND ETHANOL EXTRACTS CASUARINA EQUISETIFOLIA FOR POTENTIAL ANTIMICROBIAL ACTIVITY Uma Maheswari Narendra Kumar* and Thenmozhi Panneerselvam PG and Research Department of Microbiology, Sengamala Thayaar Educational Trust Women’s College, Mannargudi, Tamilnadu, India.

Article Received on 15 May 2015, Revised on 08 June 2015, Accepted on 01 July 2015

ABSTRACT Casuarina equisetifolia the most cultivated species of casuarinales In India. The plant is used in folk medicine for the treatment of astringent in diarrhoea, cough, ulcers, toothache, lotion for swelling and diabetes. The antimicrobial activities of leaves and bark extract were

*Correspondence for Author

investigated against medically important bacterial and fungal species,namely S.aureus ,E.coli ,Pseudomonas aeroginosa, Proteus

Dr. Uma Maheswari Narendra Kumar

vulgaris K. pneumoniae and Aspergillus niger, Aspergillus flavus,

PG and Research

Rhizopus and Fusarium. The antimicrobial activity of aqueous and

Department of

ethanol extract was determined by agar disc diffusion method. Ethanol

Microbiology, Sengamala

extracts were more active than the aqueous extract. The plant extract

Thayaar Educational Trust Women,s College, Mannargudi, Tamilnadu,

were more active against gram positive and gram negative bacteria. The most susceptible bacteria were Pseudomonas aeroginosa > K. pneumonia > Proteus vulgaris > E.coli > S.aureus and A.niger. From

India.

the screening experiment, leaves extract of Casuarina equisetifolia showed the best antimicrobial activity, hence this plant can be further subjected to isolation of the therapeutic antimicrobials and further pharmacological evaluation. KEYWORDS: Aqueous extract, Ethanol extract, Antimicrobial activity. INTRODUCTION India is the largest planter of Casuarina in the world and it is estimated that about 500,000 ha are planted with Casuarina in the states of Andhra Pradesh, Orissa, Tamil Nadu and of casuarinaceae in India, belongs to the order Casuarinales, a distinctive group of angiosperms.[1] Due something of an enigma diarrhoae, dysentery, headache and fever, cough, ulcers, toothache, lotion for swelling and diabetes. Now a day’s multiple drug

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resistance has developed due to the indiscriminate use of commercial antimicrobial drugs commonly used in the treatment of infectious disease. In addition to this problem, antibiotics are sometimes associated with adverse effects on the host including hypersensitivity immunesuppression and allergic reactions.[6] This situation forced scientists to search for new antimicrobial substances. Given the alarming incidence of antibiotic resistance in bacteria of medical importance, there is a constant need for new and effective therapeutic agents. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases from medicinal plants.[5] Therefore, the present study has been carried out to explore the antimicrobial activity of aqueous, ethanolic extract of leaves and barks of C.equisetifolia against some pathogens. MATERIALS AND METHODS Collection of plant materials The samples of C. equisetifolia leaves and barks were collected from S.T.E.T women’s college campus Sundarakkottai, Thiruvarur District in Tamil Nadu. The plant leaves and bark were washed in running tap water to remove debris and dust particles and then rinsed in distilled water for 5 minutes. It was allowed to air dried for 24 hrs. Preperation of extracts from C.equisetifolia Aqueous extract Ten gram of air-dried powder form of

leaves and bark were added to distilled water and

boiled on slow heat for 2 hrs. It was then filtered through 8 layers of muslin cloth and centrifuged at 500g for 10 minutes. The supernatant was collected, this procedure was repeated twice .After 6 hours , the supernatant collected at an interval of every 2 hrs was pooled together and concentrated to make the final volume one-fourth of the original volume It was autoclaved at 121⁰C and at 15lbs pressure and stored at 4⁰C . Then aqueous extraction was prepared 0%, 25%,50%,75%,100% concentration. Solvent extract Ten gram of dried powder

form of leaves and bark was extracted with

various

concentration of ethanol 25%, 50% ,70%,100% .It was kept on a rotary shaker for 24 hrs. Thereafter, it was filtered and centrifuged at 500g for 15 minutes. The supernatant was collected and the solvent was evaporated to make the final volume one fifth of the original volume .It was stored at 4⁰C.

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Antimicrobial assay Antimicrobial assay was done by the standard method of disc diffusion.[5] Statistical analysis For each experiment, three replicates and three determinations were conducted. Means of variable standard error and least significant differences were carried out. RESULTS The antimicrobial activity was performed by the C.equisetifolia leaves and bark .The maximum activity was shown at 100% concentration. Hence aqueous leaves extract of C.equisetifolia against S.aureus (12±0.4), E.coli(14±0.08), B.subtilis (15±0.28), P.aeroginosa (17±0.41), P.vulgaris (13±0.2) K.pneumoniae (16±0.25). A.niger(12±0.29) A.flavus (11±0.1), Fusarium (10±0.04), Rhizopus (8±0.37). Ethanol extract of C.equisetifolia S.aureus (13±0.93), E.coli (15±0.6) ,B.subtilis (180.1), P.aeroginosa (24±0.9), P.vulgaris (20±0.23), K.pneumoniae (22±0.56). Apergillus niger (14±0.29), Apergillus flavus (13±0.12), Fusarium( 12±0.04) and Rhizopus (10±0.37). In aqueous bark extracts against S.aureus (6±0.41), E.coli (8±0.08), B.subtilis (9±0.08), P.aeroginosa (10±0.25), P.vulgaris, K.pnemoniaem (11±0.41). Apergillus niger (8±0.37), Apergillus flavus (7±0.12), Fusarium (6±0.08), Rhizopus (5±0.25). Ethanolic extract of bark against S.aureus (10±0.24), E.coli (14±0.91), B.subtilis (15± 1.8), P.aeroginosa

(19±

1.75),

P.vulgaris

(18±

1.5),

K.pneumoniae

(17±

1.4).

Apergillusniger(11±0.45), Apergillusflavus (10±0.18), Fusarium(9±0.06), Rhizopus(7±0.56). Compared to aqueous extract, ethanolic extract of leaves shown better antimicrobial activity against tested pathogens (table.1). Besides the pathogen, Pseudomonas aeroginosa, K.pneumoniae, Bacillus subtilis and Aspergillus niger are effectively controlled by agar disc diffusion method.The plants extract were more active against the gram negative bacteria than gram positive bacteria. The fungal strain Aspergillus niger was effectively controlled by ethanolic extract of C.equisetifolia .Susceptibility differences between the gram positive and gram negative bacteria may be due to cell wall structural differences between these classes of bacteria.

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Table.1Antibacterial activity of aqueous extract of leaves, bark of C.equisetifolia Pathogenic bacteria Aqueous extract Zone of inhibition S.aureus 12±0.4 E.coli 14±0.08 B.subtilis 15±0.28 Leaves P.aeroginosa 17±0.41 P.vulgaris 13±0.25 K.pneumoniae 16±0.25 S.aureus 6±0.41 E.coli 8±0.08 Bark B.subtilis 9±0.08 P.aeroginosa 10±0.25 P.vulgaris 7±0.25 K.pneumoniae 11±0.41 Values are triplicates, and represented as mean ± SD Table.2 Antibacterial activity of ethanolic extract of leaves, bark of C.equisetifolia Pathogenic bacteria Ethanol extract Zone of inhibition S.aureus 13±0.93 E.coli 15±0.6 B.subtilis 18±0.1 Leaves P.aeroginosa 24±0.9 P.vulgaris 20±0.23 K.pneumoniae 22±0.56 S.aureus 10±0.24 E.coli 14±0.91 B.subtilis 15±1.8 Bark P.aeroginosa 19±1.75 P.vulgaris 18±1.5 K.pneumoniae 17±1.4 Values are triplicates, and represented as mean ± SD Table. 3 Antifungal activity of aqueous extract of C.equisetifolia leaves and bark extract Pathogenic fungi Aqueous extract Zone of inhibition A.niger 12±0.29 A.flavus 11±0.12 Leaves Fusarium 10±0.04 Rhizopus 8±0.37 A.niger 8±0.37 Barks A.flavus 7±0.12 Fusarium 6±0.08 Rhizopus 5±0.25 Values are triplicates, and represented as mean ± SD

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Table. 4 Antifungal activity of aqueous extract of C.equisetifolia leaves and bark extract Pathogenic fungi Ethanol extract Zone of inhibition A.niger 14±0.29 A.flavus 13±0.12 Leaves Fusarium 12±0.04 Rhizopus 10±0.37 A.niger 11±0.45 Barks A.flavus 10±0.18 Fusarium 9±0.06 Rhizopus 7±0.56 Values are triplicates, and represented as mean ± SD DISCUSSION Our study showed that the extract of aqueous and ethanol extracts of C.equisetifolia for potential antimicrobial activity. Aqueous and ethanol extract of C.equisetifolia leaves active against some pathogenic bacteria and fungi P.aeroginosa, B.subtilis, K.pneumoniae, E.coli ,Proteus vulgaris, S .aureus and fungi A.niger, Fusarium, A.flavus and Rhizopus. The leaves extract of C.equisetifolia was exhibited better results on P.aeroginosa, B. subtilis, K.pneumoniae, E.coli, P.vulgaris and S. aureus. Our studies are in agreement with our results are coincide with the findings of . antimicrobial efficacy of Casuarina equisetifolia extract against pathogenic organisms. The results were recorded that the tested ethanol extracts of leaf of C.equisetifolia showed good activity against Pseudomonas aeroginosa, > Bacillus subtilis > K.pneumoniae > E.coli > Proteus vulgaris >Staphylococcus aureus.[4] Our findings were similar to efficacy of aqueous and ethanol extracts of some medicinal plants for potential antimicrobial.[5] The plant extracts were more active against Gramnegative bacteria than against Gram- positive bacteria. The most susceptible bacteria were B.subtilis, followed by S.aureus, while the most resistant bacteria were P.aeroginosa, followed by K.pneumoniae. From the screening experiment, C.equisetifolia showed the best antimicrobial activity. CONCLUSION In conclusion Casuarina equisetifolia extracts possess a broad spectrum of activity against a panel of microbes responsible for the most common pathogens diseases. These promissory extracts open the possibility of finding new clinically effective antimicrobial compounds.

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Further research is necessary to determine the identity of the antimicrobial compounds from with in this plants and also to determine their full spectrum of efficacy. REFERENCES 1. Subramaniyan KN, Casuarinas Trees of Multiple Utility, Institute of Forest Genetics and Tree Breeding, Coimbatore,1992. 2. Varghese TM. An Introduction to the Anatomy of Angiosperms,Allied Publishers Limited,New Delhi, 1987. 3. Pinyopusarerk K. Range- wide provenance variation in growth and morphological characteristics of C.equisetifolia grown in Northen Australia. For. Ecol. Manage. 2000; 134:219-232. 4. Nehad M, Gumgumjee. Antimicrobial efficacy of

Casuarina equisetifolia extracts

against some pathogenic microorganisms J.of.Medicinal Plants Research, 2012; 6(47): 5819-5825. 5. Nicodemus A. Genetic improvement of C.equisetifolia through selection and breeding.Envis For. Bull. 2007; 7(1): 91-99. 6. Jigna PA, Darshana JA, Sumitra CH. Efficacy of Aqueous and Methanol Extracts of Some Medicinal Plants for Potential Antibacterial Activity. 2005; Turk J. Biol. 29: 203210. 7. Ataia A, Pal SC, Patil UK, Yadav SK. Evaluation of preliminary anticancer activity of Casuarina equisetifolia Frost (Casuarinaceae). Planta Indica. 1983; 4: 45-48. 8. Kishore DV, Rahman R. Spasmolytic activity of Casuarina equisetifolia bark extract. Int. J. Pharm. Sci. Res. 2012; 3(5): 1452-1456. 9. Rajib D, Monirul MD, Haque H, Mossaddik A. In vitro Antibacterial screening and toxicity study of some different medicinal plants. World J. Agric. Sci. 2009; 5(5): 617621. 10. Parekh J, Jadeja D, Chandra S. Efficacy of aqueous and methanolic extracts of some medicinal plants for potential antibacterial activity, Turkish J. Biol. 2005; 29: 203-210.

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