Published By Science Journal Publication Science Journal of Microbiology ISSN:2276-8572 International Open Access Publisher http://www.sjpub.org/sjarm.html © Author(s) 2014. CC Attribution 3.0 License. Research Article Volume 2014, Article ID sjmb-249, 3 Pages, 2014. doi: 10.7237/sjmb/249 Antimicrobial activity of the seed crude extracts of Triumfettarhomboidea (Tiliaceae) evis K. Mokaya¹ Josiah O. Omolo¹, Regina K. Mayaka¹, Alice W. Njue¹, John O. Adongo¹ and Joseph Matofari² ¹Chemistry Department, Egerton University, P. O. Box 536, 20115-Egerton, Kenya. ²DAFTEC Department, Egerton University, P. O. Box 536, 20115-Egerton, Kenya. Accepted 28�� May, 2014 ABSTRACT Hexane, dichloromethane and methanol extracts of the seeds of Triumfettarhomboideawere assayed for their invitro antimicrobial properties against four bacterial species:S. typhimurium, E. coli, B. subtilis, B. cereusand one yeast species- C. albicans, using agar disk-diffusion method. All the three extracts exhibited varying degrees of antimicrobial activities, with the dichloromethane extract showing activity against all the microbes tested. This study shows that T. rhomboidea, which is currently used as traditionalmedicine in certain parts of the world,is a potential source of antimicrobial compounds. development potent anti-microbial agents. Phytochemicals derived from plants have shown great promise in the treatment of many infectious diseases (Nascimento et al., 2000; Rios and Recio, 2005). These reasons justify the search for more potentially effective and new antimicrobial compounds among the many plant species (Pretorius et al., 2003, Moreillion et al., 2005). KEYWORDS: Triumfettarhomboidea,antimicrobial activity, medicinal plant. There are about seventyknown species of Triumfetta which are distributed. Triumfettarhomboidea, is still largely used ethnomedically in various parts of the globe as traditional medicine. The purpose of this study was to investigate the antimicrobial activities of seed extracts of T. rhomboideaagainst selected microbes. INTRODUCTION MATERIALS AND METHODS Triumfettarhomboidea(also known as the Chinese burr) is a perennial herb widely distributed widely across the tropical regions of the world (Royal Botanic Gardens & Domain Trust, 2010)and been used in folk medicine and therapy for the treatment of various diseases, such as gonorrhoea, diarrohea, and leprosy. Various Parts of the plant used therapeutically are fruit, flower, leaves, bark and root. Roots are used as diuretics and in the treatment of intestinal ulcer and the stem barks are used in diarrheoa and dysentry treatment and also as an aphrodisiac. It leaves, flowers and fruit concoctionsact as astringents (Chopra et. al, 1986, Charttegee, 1992, Mukharjee, 2002, Baernes, 2002, Khare, 2007). Plant material extraction procedure Infections caused by bacteria and fungi account for approximately one-half of all deaths in the developing countries. The emergence of multi-resistant microbial pathogens towards the current antimicrobial drugs and the rise in new infectious diseases still remain a serious global threat to human health (WHO, 2008). Developing countries have a rich biodiversity of plant resources that exhibit a wide range of biological activities which may help in the The seeds of T. rhomboideaplantwere collected from Rongo District in Western Kenya in August 2008, were dried to constant weight.The taxonomic identification of plant materials was confirmed by Department of Botany, Egerton University, Kenya and where a voucher specimen was deposited at the herbarium section. The collected plant seeds were air dried in the shade for 24 h and pulverized using a milling machine (MILWAUKE WISCONSIN 53207 Model No. 311). The powdered material (400 g) was first soaked 1 L of hexane for 24 hrs, and then filtered using (Whatman filter paper No. 1) to obtain the hexane extract. The above procedure was repeated in successively with the plant residue to obtain the dichloromethane and methanol extracts. The three crude filtrates were concentrated invacuousing a Rotary evaporator (BUCHI Rotavapor Model No. R-205) while maintaining the bath temperature at 40°C in order to avoid thermal decomposition of the labile compounds. The crude extracts obtained were then stored Corresponding Author: John O. Adongo Chemistry Department, Egerton University, P. O. Box 536, 20115-Egerton, Kenya. Email address: dadyjohn2004@gmail.com Science Journal of Microbiology( ISSN:2276-8572) page 2 in pre-sterilized sample vials at –4°C awaiting antimicrobial tests. Microorganisms A total of five microorganisms, one yeast species, and five bacterial species, were used as the test organisms in this study. The list of microorganisms used is given in Table 1. Pure cultures of thetest microorganisms were obtained from the Dairy and Food science Technology (DAFTEC) Department of Egerton University, Kenya. was used as a positive control and clotrimazole (30 µg/mL) was used for fungi. The inoculated plates were incubated at 37°C for 24 h for bacterial strains, and 48 h for yeast isolates after which the test plates were examined. Following incubation, the zones of inhibition formed were measured (in mm) and the mean diameter obtained to assess their respective antimicrobial activities. Each assay in this experiment was done in triplicate. Overall, cultured bacteria with halos equal to or greater than 7 mm and fungi with 10 , considered susceptible to the tested extract mm halos were (Okerulu, 2001). RESULTS AND DISCUSSION Antimicrobial activity The stored crude extracts were dissolved in their respective solvents (hexane, dichloromethane and methanol) to a final concentration of 30 mg/ml using 0.45 µm milipore filters. Antimicrobial tests were then carried out by disk diffusion method (Murray et al., 1995) using 100 µl of suspension containing 108 CFU/ml of bacteria, 106 CFU/ml of yeast spread on nutrient agar (NA) and sabouraud dextrose agar (SDA) respectively. The nutrient agar was prepared by weighing 28 g of agar in 1000 ml distilled water in a Erlenmeyer flask and then sterilized for 15 minutes at 121�C in an autoclave before being left to cool. After cooling the media was poured into the Petri dishes ready for the introduction of the microorganism. The disks (6 mm in diameter) were impregnated with 10 µl of the extracts (400 µg/disk) at the concentration of 30 mg/ml and placed on the inoculated agar. Negative controls were prepared using the sterilized paper disks without extracts or antibiotics. Ciprofloxacin was used as a standard. 20 % v/v WFI in DMSO The antimicrobial activities of Triumfettarhomboidea, (hexane, dichloromethane and methanol) extracts against the microorganisms (Candida albicans, Bacillus cereus, Salmonella typhimurium, Escherichia coli, Bacillussubtilis) were examined in the study. Themicrobe’s sensitivity, were qualitatively assessed by the presence or absence of inhibition zones, and the inhibition zone (IZ) diameters. The antimicrobial activity results containing the determined IZ values for the each test organism are reported in table 1. The results indicate that the dichloromethane extract of T. rhomboidea showed activity against all the microbes tested. The hexane extract did not show activity against C. albicans, this indicates that the anti-fungal compounds from the seeds are relatively non-polar. Escherichia coli showed no sensitivity towards both the hexane extract and the methanolic extracts. S. typhimuriumwas most sensitive to the dichloromethane extract (20.3 mm) while C. albicans was least sensitive towards the methanolic extract (8.1 mm). Table 1. Antimicrobial activity of crude extracts of Triumfettarhomboidea Inhibition zone diameter (mm) around test disk Triumfettarhomboidea extracts (400 μg/disk) Test Organisms C. albicans B. cereus S. typhimurium E. coli B. subtilis Hexane 10.2±0.8 14.3±0.7 12.4±0.6 Dichloromethane 10.1±0.3 12.4±0.4 20.3±0.5 8.9±0.3 11.4±1.2 Methanol Positive Control (STD*) 11.1±1.1 9.2±0.2 15.1±0.7 10.2±0.5 37.8±0.6 36.6±0.9 35.6±0.5 35.6±0.5 37.1±0.8 *STD - Positive control for bacterial pathogens was ciprofloxacin and for fungi, clotrimazole. N/B: All the negative controls showed no activity against the pathogens. Two gram positive bacterial strains (B. cereus, and B. subtilis) were sensitive to hexane, dichloromethane and methanol crude extracts. The gram negative (E. coli) was only sensitive to the dichloromethane extract while the gram negative (S. typhimurium) was sensitive to the hexane, dichloromethane and methanol extracts. This result suggest that the seed T. rhomboideacontains compounds sensitive to both gram positive and gram negative bacteria. The compounds responsible for the anti-microbial activity in the crude extracts from the seeds of T. rhomboideahave varied polarities between polar, mid-polar and very polar. This is because hexane is the least polar of the three solvents and methanol, the most polar. Dichloromethane is considered mid-polar. CONCLUSIONS The results of this study suggest that Triumfettarhomboidea seeds possess compounds with antimicrobial activity against fungal and bacterial pathogens. Its observed antimicrobial activity provides an explanation for its use in traditional folk medicine. Considering that the plant has beneficial How to Cite this Article: evis K. Mokaya,Josiah O. Omolo,Regina K. Mayaka,Alice W. Njue,John O. Adongo and Joseph Matofari, "Antimicrobial activity of the seed crude extracts of Triumfettarhomboidea (Tiliaceae)",Science Journal of Microbiology, Volume 2014, Article ID sjmb-249, 3 Pages, 2014. doi: 10.7237/sjmb/249 page 3 antimicrobial activities, it can be cultivated for use as a potential source of antifungal and anti-antibacterial compounds. Science Journal of Microbiology( ISSN:2276-8572) 7. Mukharjee, P.K. (2002). In, Quality Control of Herbal Drugs, 1st ed., Business Horizons, Pharmaceutical Publishers, pp. 40. 8. Murray PR, Baron EJ and Pfaller MA (1995). Manual of Chinical Microbiology6th Edition. ASM: Washington DC. pp 213-215. 9. Nascimento, G.G.F., Locatelli, J., Freitas, P.C., and Silva, G.L. (2000). Antibacterial activity of plant extracts and phytochemcals on antibiotic resistant bacteria. Brazillian Journal of Microbiology.31: 247-256. ACKNOWLEDGEMENTS We wish to thank technical staff of Chemistry and DAFTEC Departments of Egerton University, Kenya for their technical assistance. REFERENCES 1. Barnes, A. (2002). An Introduction to Herbal Medicinal Products, The Pharmaceutical Journal: 268, pp. 804. 2. Chattergee A, Chandra Prakash S. (1992). National Institute of Science and Communication, CSIR, New Delhi, The Treatise of Indian Medicinal Plant:2. pp. 170-171. 3. Chopra R., Nayar S., and Chopra, I. (1986). In; Glossary of Indian Medicinal Plant, 3rd Ed., Council of Scientific and Industrial Research, New Delhi, pp. 249-250. 4. Khare, C. P. (2007). Indian Medicinal Plants, Springer Publications, USA, 2007, pp.-677. 5. Mevy J.P., Bessiere J.M., Rabier J, Dherbomez M, Ruzzier M, Millogo, J, and Viano J. (2005), Composition and antimicrobial activities of the essential oil of Triumfettarhomboidea. Flavour and Fragrance Journal.21: 80 - 83. 6. Moreillion, P, Que, Y. A and Glauser, M. P. (2005). Staphylococcus aureus (Including Staphyloccal Toxic shock). In 'Principles and Practice of Infectious diseases. In Mandell G.L, Bennett J.E, Dolin R. Published by Churchill livingstonePennyslyvania 6th ed.: 2: 2333- 2339. 10. Okerulu, I. O. (2001). The phytochemical analysis and antibacterial screening of extracts of Tetracarpidiumconophorum. Journal of Chemical Society. (Nigeria).26:223-228. 11. Pretorius, J. C, Magama, S and Zietsman, P. C. (2003). Growth inhibition of plant pathogenic bacteria and fungi by extracts from selected South African plant species. South African Journal of Botany.20: 188-192. 12. R´?os, J.L., and Recio, M.C. (2005). Medicinal plants and antimicrobial activity Journal ofEthnopharmacology. 100: 80-84. 13. Royal Botanic Gardens & Domain Trust. (2010). "Genus Triumfetta". PlantNET - New South Wales Flora Online., Sydney Australia. Available at http://eol.org/data_objects/26754058#cite_note-NSW-2 14. World Health Organization (WHO). (2001). Traditional medicine. Fact sheet number 134. Revised May, 2003. Availableat: http/www.who.int/media centre fact sheet/fs/134. Accessed on 2nd March, 2014. How to Cite this Article: evis K. Mokaya,Josiah O. Omolo,Regina K. Mayaka,Alice W. Njue,John O. Adongo and Joseph Matofari, "Antimicrobial activity of the seed crude extracts of Triumfettarhomboidea (Tiliaceae)",Science Journal of Microbiology, Volume 2014, Article ID sjmb-249, 3 Pages, 2014. doi: 10.7237/sjmb/249