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Ethnobotanical
Leaflets 12: 570-76. 2008. Antimicrobial and Cytotoxic Activities
of Hopea utilis Fruits M. Maridass Environmental Carcinogenesis Research Unit St. Xavier’s College (Autonomous), Palayamkottai - 627 002 Tamil Nadu, Corresponding Email address: orchideyadass@yahoo.com Issued Abstract Aqueous and ethanolic crude extracts of Hopea utilis screened for antibacterial and cytotoxic activities were studied. Antibacterial activity of ethanolic extracts of H. utilis were more successful with the pathogens Salmonella typhi and Streptococcus aureus. The MICs values of ethanolic extract of were Hopea utilis active against Salmonella typhi (25mg/ml), Salmonella typhi (25mg/ml), and Staphylococcus aureus (36mg/ml) respectively. The results of both extracts of aqueous and ethanolic of Hopea utilis showed the brine shrimp lethality assay LD50 values were 1.64μg/ml and 1.34μg/ml. Key Words: Hopea utilis;
fruits; antibacterial activity; ethanolic extract. Introduction Plants
are recognized for their ability to produce a wealth of phytochemicals.
Humankind has for centuries used many species to treat several diseases
(Cragg et al., 1999). Recently, a number of tribal studies concerning the
search for new antimicrobial agents from plants and antimicrobial screening
of the extracts have been published.
A rapid and
inexpensive test, brine shrimp (Artemia salina) (BST), has been used
for screening of biological and cytotoxity activities (De Rosa et al., 1994).
The fractions or active compounds in this assay, are further tested in
cultured tumoral cells, antimicrobial and antiparasitic assays, generally
with good correlation (Sahpaz et al., 1994; Colman-Saizarbitoria et
al., 1995; Siqueira et al., 1998). The Dipterocarpaceae is a plant family of 14
genera 750 species found throughout the tropical and temperate regions of the
world. Bandaranayake et al., (1977) reported that 44 of the 45 species
are endemic. They belong to the genera Cotylelobium, Hopea, Dipterocarous,
Shorea, Stemonoporus, Vateria and Vatica. Hopea utilis is a large
sized tree distributed in evergreen
forest, Southern Western Ghats region, up to
sea level 5000 meters. Hopea utilis is locally known as ‘‘Karapongu’’
in Karaiyar region. Ethnobotanical information gathered from tribe of Kanis
used as fruits was boiled with water and
treatment of stomach pain and hypertension. Bioactive constituents of
hopeafuran and C-glycosyl resveratrol were isolated from the stem wood
(Tanaka et al., 2001). In
the present study, the in - vitro antibacterial and cytotoxic activities of
aqueous and ethanol extract of Hopea
utilis fruits were investigated. Materials and Methods Collection of plant materials The fruits materials of H. utilis were collected in the Karaiyar, Tirunelveli District, South India and was identified by Dr. U. Manikandan, SPKCES, Alwarkurichi, South India. Extract preparation 250gm of powdered materials was individually extracted with 95% ethanol and water at room temperature. The solvent extract was removed by distillation method. The resulting crude extracts were stored at -20°C until assayed. Antibacterial screening Test Organisms Bacillus
subtilis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus
pyogenes, Vibrio paarahaemolyticus and Salmonella typhi were studied. Inocula Inocula
for the bioassays were prepared by diluting scraped cell mass in 0.85% NaCl
solution, adjusted to McFarland scale 0.5 and confirmed by spectrophotometrical
reading at 580nm. Cell suspensions were finally diluted to 104 Disk-
diffusion method In - vitro antibacterial
activity of both extract, were studied against four gram - positive and two
gram - negative bacterial strains by the standard disc-diffusion method
(Barry , 1980; Buer et al., 1966; Berghe and Vlientinck, 1991).
Nutrient agar was the bacteriological medium. Both extracts were screened at
a concentration of 100µg /ml. Diameters of zones of inhibition produced by the
isolated agent were compared with those produced by the standard antibiotic
(Kanamycin, 30µg /disc ). Determination of MIC and MBC Minimal inhibitory concentration (MIC) and
Minimal bactericidal concentration (MBC) were determined for the extracts detailed
methods (Suffredini et al., 2004) Cytotoxic activity The
cytotoxic effect of both, aqueous and ethanolic extract was evaluated by LC50
of brine shrimp lethality test were followed by Mayer et al.,
1982 and Persoone, 1980. Both aqueous
and ethanolic extract were dissolved in dimethylsulphoxide (DMSO)
separately and five graded doses 5, 10, 20, 40 and 80 mg ml/L were used for 5ml
sea water containing 10 brine shrimp nauplii in each group. Mortality was recorded
12h and dead animals were removed immediately. LC50 values, upper
and lower confidence limits and slope values were calculated using the POLO
computer software (Russell et al.,
1979). The regression coefficient between exposure time and different values
of LD50 was determined by probit analysis (Finney, 1947). All
computations were performed with a computer with a capacity of 28GB in the
hard drive, 256 MB in RESULTS Many microorganisms which cause damage to human health exhibit drug resistance due to inadequate use of antibiotics. Thus, there is a need for the discovery of new substances from natural resources, including plants (Sartoratto et al., 2004). In the present study, the antibacterial activity of aqueous and ethanolic extract of H. utilis is shown in Table 1. Antibacterial activity of ethanolic extract of H. utilis was more activity observed both pathogen of Salmonella typhi (30mm) and Streptococcus aureus (29mm). The MICs values of ethanol extract active against Salmonella typhi 25mg/ml, and Staphylococcus aureus 36mg/ml, respectively. The cytotoxicity of the compound was bioassayed against brine shrimp nauplii and the results were shown in Table 2. The 50% mortality of log-dose concentrations (LD50) of the aqueous extract was 1.62 µg/ ml. Figure- 1 shown the 95% confidence regression value Y=3.67+1.44X and significant level of (P< 0.05) were observed in aqueous extract of H. utilis. While the more cytotoxic activity was observed in 95% ethanol extract observed seen in Table-2 and Fig.2. The present study was agree to previous workers studied in bioactive compounds of Kolavenic acid and Clerodane diterpine (Islam et al., 2001); Isoflavone (Shah Alam Bhuyan et al., 2003); Triterpenoid (Rahman et al., 2002) and galic acid (Saker et al., 1998). The antibacterial and cytotoxic activity of extract of H. utilis bioassay-guided fractionation procedure to characterize and isolate the bioactive principle is under way in our laboratory. Acknowledgements The author is grateful to Principal, St.Xavier’s College (Autonomous), Palyamkottai-627002, providing for laboratory facilities. REFERENCES Ahmad, I.; Beg, A.Z. Antimicrobial and phytochemical studies on 45 Indian plants against multi-drug resistant human pathogens. J. Etnopharmacol., 74:113-123, 2001. Ali-Shtayeh, M.S.A.; Yaghmour, R.M-R.A.; Faidi, Y.R.B.; Salem K.; Al-Nuri, M.A.D. Antimicrobial activity of 20 plants used in folkloric medicine in the Palestinian area. J. Ethnopharmacol., 60:3: 265- 271, 1998. Baba-Moussa, F.; Akpagana, K.; Bouchet, P. Antifungal activities of seven West African Combretaceae used in traditional medicine. J. Etnopharmacol., 66:3:335-338, 1999. Lentz, D.L.; Clark, A.M.; Hufford, C.D.; Meurer-Grimes, B.; Passreiter, C.M.D.; Cordero, J.; Ibrahimi, O.; Okunade, A.L. Antimicrobial properties of Honduran medicinal plants. Short communication. J. Ethnopharmacol., 63:253-263, 1998. Cragg GM,
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1999. International collaboration in drug discovery and development: the Mahasneh, A.M.A.; Adel, M.A.; El-Oqlah, A.A.B. Antimicrobial activity of extracts of herbal plants used in the traditional medicine of Jordan. J. Ethnopharmacol., 64:271-276, 1999. Martínez, M.J.; Betancourt, J.; Alonso-González, N.; Jauregui, A. Screening of some Cuban medicinal plants for antimicrobial activity. J. Ethnopharmacol., 52:171-174, 1996. Panizzi, L.; Flamini, G.; Cioni, P.L.; Morelli, I. Composition and antimicrobial properties of essential oils of 4 mediterranean Lamiaceae. J. Ethnopharmacol., 39:167-170, 1993. Russell, R.M., Robertson, J.L., and Savin, N.E. (1977). POLO: A new computer programme for probit analysis. Bulletin of the Entomological Society of America, 23: 209-213. Sartoratto,A., Machado,A.L.M., Delarmelina,C., Figueira,G.M., Cristina,M., Duarte,T., Rehder,V.L.G.2004. Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil. Brazilian Journal of Microbiology, 35 : 275-280 Singh, D.K., and Agarwal, R.A. (1984). Correlation of the anticholinesterase and molluscicidal activity of the latex of Euphorbia royleana Bioss. on Lymnaea acuminata. Journal of Natural Products, 47: 702-705. Sokal, R.R.,and Rohlf, F.J. (1973). Introduction to Biostatistics. W.H. Freeman and Co., San Francisco, 271-273. Table 1. Antibacterial activity of ethanolic and aqueous extract of H. utilis fruits.
Table 2. Antibacterial activity of ethanolic and aqueous extract of H. utilis fruits.
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