A Comparative Study of Antimicrobial Effects of Clitoris ternatea and Anacardium occidentale Leaf Extracts Against the Selected Microbes

: The aim of the present study was to investigate a comparative study of antimicrobial efficacy of Clitoris ternatea and Anacardium occidentale leaf extract against Staphylococcus aureus, Streptococcus pneumoniae, Mycobacterium tuberculosis and Bacillus subtilis (Gram-positive) and Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus vulgaris and Salmonella typhi (Gram-negative) microbial strains. Ethanol extracts of both the leaf were evaluated against the experimental microbial strains using the well diffusion and agar disc diffusion methods. The present investigation showed that some microbial strains were more sensitive to Clitoris ternatea as compared to Anacardium occidentale and vice versa. But on the whole the Clitoris ternatea was found to be more effective than Anacardium occidentale. It is concluded that both the extracts showed antimicrobial activity due to the presence of bioactive phytochemicals present in them.


INTRODUCTION
Several bioassays such as disc diffusion, well In the recent past decades, the antimicrobial commonly used.To promote the proper use and to activities of plants have been investigated by a determine the potential of plants as a source of new number of researchers worldwide and they have drugs, it is essential to understand their properties, safety enormous therapeutic potentials for incurable diseases and efficacy as the first and foremost priority.[1].The bioactive compounds of plants have a wide Clitoris ternatea Linn (Fabaceae) is a twining climber range of biological functions including antimicrobial, found abundantly in tropical and subtropical regions, antioxidant, anti-inflammatory activities, etc.The mostly as terrestrial mesophytes which are commonly development of pharmaceutical drugs derived from plants used in Indian folk medicine.It is popularly known as a " has allowed the treatment of diseases on a large effective Butterfly pea" in western countries and as Aparajita in the scale.The consumption of antimicrobial drugs may result traditional Ayurvedic system of medicine.It is employed in unfavorable and untold side effects.Plants accumulate in the traditional system of medicine against different various secondary metabolites including alkaloids, diseases such as cathartic, purgative, demulcent, emetic glycosides, polyphenol, etc [2].They are effective in the and anti-inflammatory [5].Various parts of it have been treatment of infectious and incurable diseases with reported to have nootropic, anxiolytic, tranquilizing, minimal side effects and are associated with synthetic analgesic, antipyretic, antimicrobial and antimicrobials.The usage of drugs has a negative effect immunomodulatory activities.It is reported to have brain on the human body and led to the concurrent resistant ionic activity and is popularly known as Shankhapushpi bacteria that has become a global issue of public health.
in India [10].Therefore, it is worthwhile to evaluate its Researchers are engrossed in developing new antimicrobial effects on the selected microorganisms.antimicrobial agents from various sources to combat Anacardium occidentale Linn (Anacardiaceae) a microbial resistance.Therefore, greater attention has been native of Brazil, having great economic and medicinal paid to antimicrobial activity, screening and evaluating value, which is composed of about 74 genera and methods.600 species.The leaves, stem and bark extract are utilized diffusion and broth or agar dilution are well known and widely for the treatment of diarrhoea, dysentery and Microbial Strains Used: Staphylococcus aureus ATCC colonic pain [3].It has also been reported to possess 25923, Streptococcus pneumonia ATCC 29212, anti-diabetic, anti-bacterial, anti-inflammatory and Mycobacterium tuberculosis MTCC 12556 and Bacillus anti-ulcerogenic [4].The leaves are also used in Brazil for subtilis UC 564 (Gram-positive) and Pseudomonas eczema, psoriasis, scrofula, dyspepsia, genital problems, aeruginosa ATCC 25619, Klebsiella pneumonia MTCC venereal diseases, as well as for impotence, bronchitis, 109, Proteus vulgari UC 726 and Salmonella typhi ATCC cough, intestinal colic, Leishmaniasis and Syphilis related 12600 (Gram-negative) microbial strains were procured skin disordes.The nut oil is used topically as an from the microbial type collection India.antifungal and for healing cracked heels.Fruit juices from the cashew apple and leaves are known to have Antibiotics Used: Gentamicin (HIMEDIA Gentamicin GEN antimicrobial, anti-inflammatory, astringent, diuretic, 10 susceptibility test disc 10µg/disc) and Chloramphenicol hypoglycemic and other medicinal properties.The leaves (HIMEDIA Chloramphenicol c30 susceptibility test disc of which contain pharmacological rich components 10µg/disc), were used as reference standards for the such as alkaloids, essential oils, tannins, saponin, antibacterial study.cardenolides and are known to have cosmetic, antimicrobial and antioxidant properties [11].The plant Antimicrobial Assay: The ethanolic extracts of Clitoris Anacardium occidentale is very useful, as found by the ternatea and Anacardium occidentale were screened for above-mentioned reports and there is a need to find out their in vitro antimicrobial activity in comparison with more about the potentiality of this plant as an experimental standard antibiotics by the well diffusion antimicrobial agent against the selected microbial strains.
method [12].For this activity, a nutrient agar obtained

MATERIALS AND METHODS
plates were prepared by pouring 15-20 ml of molten media Plant Material: The plants Clitoris ternatea and 24 hours of bacterial culture spread throughout the Anacardium occidentale collected from the outskirts of medium uniformly with a sterile cotton swab.With the Hyderabad district Telangana state.The plant specimens help of a sterile cork, borer wells were made with a were identified and the voucher specimens deposited in diameter of 6mm at equidistant in the medium.The extracts the department of Botany.
(10, 20, 30, 40 mg/ml) were loaded into the wells and the The leaves, washed thoroughly many times with compound was allowed to diffuse for 2 hours and the running water and then finally sterile water.They are dried plates were kept in incubation at 37°C for 24 hours.in the shade at room temperature for more than three The sensitivity of the organisms was determined by weeks.Dried leaves were made into coarse powder and measuring the average diameter of triplicates of the zone sieved for crude extraction with ethanol as a solvent.
of inhibition in mm with the help of a transparent ruler [7].

Extraction:
The fine powder was made by pulverizing with performed by the agar disc diffusion following Kirby clean pestle and mortar and packed in a sealed plastic Bauer method at the concentration 10,20,30,40 mg/ml of bottle until extraction.About 20 grams of dried powder the extracts in DMSO [6].Gentamicin and were mixed with 350 ml of ethanol (5.7%) and extracted by Chloramphenicol discs were used as reference control and the hot percolated method by the Soxhlet apparatus.
pure solvent control was also maintained throughout the The extracts were transferred into a 500 ml each conical experiment.flask, tightly closed and left for 24 hours at room temperature.

RESULTS
The collected extracts, concentrated at 40°C under reduced pressure rotary evaporator to remove as much of Table (1) reveals the antimicrobial activity of the ethanol as possible.The dried extract was then mixed ethanolic extracts of Clitoris ternatea and Anacardium with dimethylsulfoxide (DMSO).At the end of the occidentale.The mean values of the zone of inhibition of procedure, the extracts were placed in a sterile glass flask, different concentrations against the test organisms sealed with paraffin and then stored at 4°C until further were compared with the standard drugs.The results of the use.
zone of inhibition manifest that the extract possesses from Himedia Laboratories Ltd Mumbai.The nutrient agar into sterile Petri plates, they were allowed to solidify.
Extracts showed more pronounced activity to gram-positive than gram-negative.The reason is attributed to the sensitivity difference between grampositive and gram-negative bacteria.This is mainly due to the morphological constitution between these microorganisms.These observations are more likely to be the fact that an outer membrane in gram-negative bacteria, which acts as a barrier to many environmental substances including antibiotics.The significant differences in the antimicrobial effects of both the extracts may be due to the molecular size of the phytochemical compound present in the extracts.

DISCUSSION
The major cause of morbidity and mortality, are infectious diseases worldwide.The number of multidrug-resistant microbial strains and the appearance of strains that reduced susceptibility to antibiotics are continuously increasing.Such an increase has been attributed to the indiscriminate use of broad-spectrum antibiotics in various infectious diseases.This situation provided the impetus to the search for new antimicrobial substances from the extracts of various medicinal plants to combat against antimicrobial resistance and for therapeutic treatments.
India has a rich heritage of using medicinal plants in traditional medicines such as Ayurveda, Siddha, Unani besides folklore practices.Medicinal plants considered as clinically effective and safer alternatives to synthetic antibiotics.The microorganisms employing new mechanisms to survive [8].From the earlier studies, it is obvious that most of the phytochemicals differ significantly in their structures and properties.These differences may be attributed to the differences in the cell wall constituents of bacteria which vary among the gram-positive and gram-negative bacteria [9].

CONCLUSION
Based in the preliminary study, the authors concluded as follows: Anacardium occidentale has a significant antimicrobial activity against the selected microbial strains which may be due to the presence of active phytochemicals in the extracts.
From the present study, it is evident the susceptibility to the extracts were more pronounced in gram-positive as compared to gram-negative organisms.This is of the opinion of authors which may be due to the structural configuration of an outer membrane in gram-negative bacteria which acts as a barrier to many phytochemicals present in the extract and the different sizes of organic phytochemical molecules.
The antimicrobial activity observed more pronounced in Clitoris ternatea than Anacardium occidentale this may be due to an absence of more bioactive molecules in the extract of Anacardium occidentale.
Further, research has to be conducted at the cellular level of the organisms for a wider understanding of the inhibition mechanism.Identification and isolation of specific antimicrobial compounds in their purified forms can pave better ways to control drug-resistant strains of microbes.The results were in accordance with the earlier researchers.

Table 1 :
Results of antimicrobial activity