Available online at www.jpsscientificpublications.com Volume – 2; Issue - 1; Year – 2016; Page: 443 – 452 Indo – Asian Journal of Multidisciplinary Research (IAJMR) ISSN: 2454-1370 PHYTOCHEMICAL PROFILES, ANTIBACTERIAL AND ANTIFUNGAL ACTIVITY OF LEAVES FROM THE Psydrax dicoccos (Gaertn) D. Umaiyambigai*, K. Saravanakumar and G. Adaikala Raj, Department of Botany, Annamalai University, Annamalai Nagar – 608 002, Tamil Nadu, India. Abstract The phytochemical analyses and antimicrobial activity of Psydrax dicoccos leaves were extracted successively with different solvents viz., petroleum ether, chloroform, ethyl acetate and methanol and screened for their antimicrobial activity against Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Vibrio cholerae, Candida albicans, C. parapsilosis and C. tropicalis by using Disc diffusion method. Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were also determined in the present research. The methanol extract of Psydrax dicoccos showed the highest antimicrobial activity against all the bacterial and fungal strains tested than the other extracts. The mean zones of inhibition produced by the extracts in Agar diffusion assays against the tested bacterial strains was ranged from 7.0 to 22.5 mm. The MIC values were between 125 and 250 μg/ml, MBC values 250 and 1000 μg/ml and MFC 500 and 1000 μg/ml values were recorded. Phytochemical analyses of different extracts of Psydrax dicoccos leaves was analyzed. The methanol extract of Psydrax dicoccos leaves showed the presence of strong phytochemicals viz., steroids, tannins, phenolic compounds and terpenoids compared to other extracts. The highest mean of zone inhibition (22.5 mm) was observed in the methanol extract of Psydrax dicoccos against the bacteria Staphylococcus aureus. From the above findings, we concluded that the methanol extract of Psydrax dicoccos will be needed to isolate and characterize the compounds. Key words: Antimicrobial activity, Psydrax dicoccos, MIC, MBC and MFC. 1. Introduction Psydrax dicoccos belongs to the family was used externally for rheumatic pains (Neelima Rubiaceae and commonly called Ceylon boxwood et al., 2011). and Nazhuvai, Irambarathaan in Tamil and Microbial infections are major public distributed in all parts of tropical and sub-tropical health problems in the developed countries. region of India. All parts of the plant have been Antibiotics are used to treat these infections. Due recognized to have medicinal properties. Plant to indiscriminate use of commercial antibiotics, possesses antipyretic activity. In India, bark was the incidence of multiple antibiotic resistances in used as febrifuge and also applied as plasters. The human pathogens is increasing. This has forced decoction of roots was used internally for treating the scientists to search for new antimicrobial diarrhoea. Bark powder boiled with sesame oil substances from various sources like medicinal plants. Medicinal plants constitute the main source of new pharmaceuticals and health care products *Corresponding author: D. Umaiyambigai (Ivanona et al., 2005). The use of traditional E. mail: [email protected] medicines was widespread in India (Jeyachandran, Received: 10.12.2015; Revised: 17.12.2015; Accepted: 25.12.2015. et al., 2007). © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Umaiyambigai/ Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(1): 443 – 452 444 Infectious disease caused by bacteria, have become one of the great challenges for viruses, fungi and parasites are still a major threat modern healthcare. Majority of scientists define to public health, despite the tremendous progress MDR as ˝resistance to at least 3 classes of in human medicine (Cosa, 2006). The past three antimicrobial agents˝ (Falagas et al., 2006). decades have seen a dramatic increase in Antimicrobial agents have substantially reduced microbial resistance to antimicrobial agents the threat posed by infectious diseases over a (Chopra, 1996). Such situation stimulates the period of time since their discovery in the 1940’s development of new anti-microbial agents in order (Lewis and Ausubel, 2006). However, the to treat the infectious disease in an effective escalation of multidrug resistance in bacteria in manner. So, this matter continued to an era to recent years has seriously jeopardized these gains. identify the potential antimicrobial agent from the This has gained worldwide attention due to the natural resources. The edible plants that used for high impact on public health. Increased usage of traditional medicine contain a wide range of antimicrobial agents to treat bacterial infections substance that can be used to treat abundant of has lead to the emergence of MDR strains infectious disease with reduced side effects (Bonnet, 2004). Invasive fungal infections are of (Duraipandiyan, 2006). In Asia, the use of great concern for human beings because they are medicinal plants to cure specific illness has been associated with unacceptable high mortality rates. use for many years (Bhattacharjee, 1998). More than 90 % of all reported fungal - related Furthermore, Malaysia is rich in various edible deaths result from species that belong to one of plants with diverse biological & pharmacological three genera: Cryptococcus, Candida and properties (Yoga, 2005). Aspergillus. In turn, superficial infections of the skin and nails are the most common fungal The numbers of resistant strains of microbial pathogens are growing since penicillin diseases in humans, affecting C. albicans 25 % of resistant and multiresistant Pneumococci caused a the population worldwide (Karan, 2009). Hence, major problem in South African hospitals in 1977. present study was carried out to evaluate the (Berkowitz, 1995) calls the emerging of drug phytochemical properties and antimicrobial resistant bacteria a medical catastrophe. Leggiadro activity of petroleum ether, chloroform, ethyl (1995) stated that effective regimens may not be acetate and methanol extracts of leaves of Psydrax available to treat some Enterococcal isolates and dicoccos against bacterial and fungal strains. that it is critically important to develop new 2. Materials and methods antimicrobial compounds for these and other Collection of Plant material and preparation of organisms before we enter the post-antibiotic era. extracts New compounds inhibiting microorganisms such as benzoin and emetine have been isolated from The fresh leaves of Psydrax dicoccos plants (Cox, 1994). The antimicrobial compounds were collected from Authukurichi region from plants may inhibit bacteria by a different (11°24′37″N 79°21′4″E), Ariyalur District, Tamil mechanism than the presently used antibiotics and Nadu, India. During the months from March to April may have clinical value in treatment of resistant 2014, the specimens were deposited in the Herbarium microbial strains. A number of traditional natural of Department of Botany, Annamalai University, products have been increased and much work has Annamalai Nagar. Collected leaves were initially been done on selected ethno medicinal plants for washed with water, then surface sterilized with antibacterial activity against pathogenic strains of disinfectant solution of 10 % sodium hypochlorite Gram negative and Gram positive bacteria (Singh, solution and finally rinsed with sterile distilled 2002). water and shade dried under room temperature and grounded in to a coarse powder. One hundred Bacterial infections especially those grams of coarse powder was extracted with caused by multi-drug resistant (MDR) bacteria different organic solvents like non-polar to polar © 2015 Published by JPS Scientific Publications Ltd. All rights reserved Umaiyambigai/ Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(1): 443 – 452 445 viz., Petroleum ether, chloroform, ethyl acetate containing 106 CFU/ml were swabbed on the top and methanol for 8 hours using Soxhlet apparatus of the solidified media and the plates were dried and the solvents were evaporated under vacuum in and uniformly spread. After drying, the extracts a rotary evaporator (Heidolph, Germany) and the were placed on the disc with sterile forceps and dried powder was stored at 4˚C for further use. gently pressed to ensure the contact with the incubated agar surface. Ciprofloxacin (5 µg/disc) Phytochemical analysis The solvents such as, petroleum ether, for bacteria and Amphotercin-B (100 units/disc) for chloroform, ethyl acetate and methanol extracts of yeast were used as positive control. Ten per cent leaves of Psydrax dicoccos were used for DMSO was used as blind control. Finally, the qualitative phytochemical analyses. Various inoculated plates were incubated at 37 °C for 24 bioactive phytochemicals namely, flavonoids, hrs. The zone of inhibition was observed and tannins, steroids, glycosides, saponins, phenolic measured in millimeters. Each experiment was compounds, terpenoids and alkaloids were carried out in triplicates. analyzed according to described by Harborne Minimum Inhibitory Concentration (MIC) (1983) and Trease (1983). Minimum inhibitory concentrations of plant different extracts were tested in MHB for bacteria Microorganisms The seven clinical bacterial isolates and and SDB for fungi described by Ericsson and three fungal species were obtained from Raja Sherris (2002). The extracts were dissolved in 10 % Muthaiah Medical College