IN VITRO AND IN SILICO ANTIBACTERIAL ACTIVITIES OF Syzgium aromaticum ESSENTIAL OIL AGAINST BACTERIA ASSOCIATED WITH OTITIS MEDIA IN CHILDREN
Keywords:Otitis media, Clove, Antibacterial activity, Essential oils, MIC, In Silico
Background: This study investigated the efficacy of the essential oil (EO) of Syzgium aromaticum L. (clove) on the bacteria associated with otitis media.
Materials and Methods: Ear swab samples were collected and bacteria isolated were identified using morphological and biochemical procedures. Essential oil was extracted from the dried flower buds using the hydro-distillation method while physicochemical and phytochemical analysis was done on the oil. Antibiotic susceptibility test and agar well diffusion was used to determine the susceptibility of the isolates to the EO. In – silico analysis was conducted to determine the drugable compound in the EO.
Results: Phytochemical analysis of the oil indicated the presence of flavonoids, sterols, phenols, carbohydrates and alkaloids. Physicochemical test of the EO exhibited the presence of eugenol (80.98%) as the most abundant phytocompound. Percentage occurrence of the bacterial isolates are as follows; Proteus mirabilis (31.25%), Staphylococcus aureus (25%), Pseudomonas aeruginosa (18.25%), Proteus vulgaris (3.12%), Moraxella catarrhalis (12.5%), Klebsiella pneumoniae (3.12%) and Staphylococcus epidermidis (6.25%). Zones of inhibition were within the range of 11.5±0.71mm-23.0±2.83mm. In silico studies demonstrated that 16 out of 38 compounds identified passed the ADMET analysis. Various compounds had different binding energies, such as linalool, carvacrol for S. aureus (2NOJ), gamma-eudesmol, eudesmol for Proteus mirabilis (6H2L), eucalyptol, gamma-eudesmol and eudesmol for Proteus vulgaris (4MCX) and Staphylococcus epidermidis (4HBL).
Conclusion: This study shows the potency of clove EO as an antibacterial agent and its component as potential lead molecules in drug development and design to combat multi – drug resistance.
Abdullah, B. H., Hatem, S. F. and Jumaa, W. (2015). A Comparative Study of the Antibacterial Activity of Clove and Rosemary Essential Oils on Multidrug Resistant Bacteria. UK Journal of Pharmaceutical and Biosciences, 3(1): 18-22.
Aduda, D. S., Macharia, I. M., Mugwe, P., Oburra, H., Farragher, B., Brabin, B. and MacKenzie, I. (2013). Bacteriology of chronic suppurative otitis media (CSOM) in children in Garissa district, Kenya: a point prevalence study. International Journal of Pediatric otorhinolarynology, 77: 1107-1111.
Adukwu, E. C., Allen, S. C. and Phillips, C. A. (2012). The anti-biofilm activity of lemongrass (Cymbopogon flexuosus) and grapefruit (Citrus paradisi) essential oils against five strains of Staphylococcus aureus. Journal of Applied Microbiology, 113(5):1217–1227.
Ahmed, W. (2016). Monitoring antioxidant and antityrosinase activity of clove aromatic flower buds. Journal of Medicinal Plants Studies, 4(2): 163-169.
Ajayi, i. A., Jonathan, S. G., Adewuyi A. and Oderinde, R. A. (2008). Antimicrobial screening of essential oil of some herbal plants from Western Nigeria. World Appied Sciences Journal, 3(1): 79-81.
Akinpelu, D. A. and Onakoya T. M. (2006). Antimicrobial activities of medicinal plants used in folklore remedies in south-western Nigeria. African Journal of Biotechnology, 5(11): 1078-1081.
Akinpelu, D. A., Odewade, J. O., Aiyegoro O. A., Ashafa, O. T., Akinpelu, O. F. and Agunbiade, M. O. (2016). Biocidal effects of stem bark extract of Chrysophyllum albidium G. Don on vacomycin-resistant Staphylococcus aureus. BMC Complementary and Alternative Medicine 16: 105.
Ako-Nai, A. K., Oluga, F. A., Onipede, A. O., Adejuyigbe, E. A. and Amusa, Y. B. (2002). The characterization of bacterial isolates from acute otiti media in Ile-Ife, Southwestern Nigeria, Journal Tropical Pediatrics, 48: 15-23.
Amel Ali Sulieman, Fadwa Mutaseim Eltayeb, Smah Ahmed Sulieman, and Nazar Abdalazeem Osman. (2015). Antimicrobial Activity of Zingiber officinale (Ginger) Oil against Bacteria Isolated from Children Throat. International journal of microbiology, 1: 1-6.
Anwar, F., Ali, M., Hussain, A. I., and Shahid, M. (2009). Antioxidant and antimicrobial activities of essential oil and extracts of fennel (Foeniculum vulgare Mill.) seeds from Pakistan. Flavour and Fragrance Journal, 24(4):170-176.
Arroll, B. (2005). Antibiotics for upper respiratory tract infections: An Overview of Cochrane Reviews." Respiratory Medicine, 99 (3): 255-261.
Bakkali, F., Averbeck, S., Aberbeck, D. and Idaomar, L. (2008). Biological effects of essential oils - A review. Food and Chemical Toxicology, 46(2): 446-475.
Benzie, I. F. and Strain, J. J. (1999). Ferric reducing/antioxidant power assay: direct measure o total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods of Enzymology, 299: 15-27.
Blois, M. S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181: 1199-1200.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254.
Brand-Williams, W., Cuvelier, M. E. and Berset, C. (1995). Use of free radical method to evaluate antioxidant activity. Food and Science Technology, 28: 25-30.
Briozzo, J. (1989). Antimicrobial activity of clove oil dispersed in a concentrated sugar solution. Journal of Applied Bacteriology, 66: 6975.
Casey, J. R. and Pichichero, M. E. (2004). Changes in frequency and pathogens causing acute otitis media in 1995-2003. Infectious Diseases Journal, 23(9): 824-828.
Chaieb, K., H. Hajlaoui, T. Zmantar, K.A.B. Nakbi, M. Rouabhia, K. Mahdouani and A. Bakhrouf. (2007a). The chemical composition and biological activity of essential oil, Eugenia cryophyllata (Syzygium aromaticum L. Myrtaceae): a short review. Phytotherapy Research., 21(6): 501-506.
Clinical and Laboratory Standards Institute (CLSI), (2010). "Performance standard for antimicrobial susceptibility testing," presented at the Approved Standard M100-S20. 30(1). National Committee for Clinical Laboratory Standards, Wayne, PA. USA.
Cripps, A. and Kyd, J. (2003). Bacterial otitis media: current vaccine development strategies. Immunology and Cell Biology, 81(1):46–51.
Dahiya, P. and Soni, A. (2014). Phytochemical analysis, antioxidant and antimicrobial activity of Syzygium caryophyllatum essential oil. Asian Journal of Pharmaceutical and Clinical Research, 7(2): 202-205.
Devi, K. P., Sakthviel, R., Nisha, S. A., Suganthy, N. and Pandian, S. K. (2013). Eugenol alters the integrity of cell membrane and acts against the nosocomial pathogen Proteus mirabilis. Archives in Pharm arceutical Research, 36:282-292.
Dickson, G. (2014). Acute Otitis media. Primary Care, 41: 11-18.
Dorman, H. J. and Deans, S. G. (2000). Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88(2): 308-316.
Egbe, C., Mordi, R., Omoregie, R. and Enabulele, O. (2010). Prevalence of Otitis Media in Okada Community, Edo State, Nigeria. Macedonian Journal of Medical Science, 3(3):299-302.
Fu, Y., Zu, Y., Chen, L., Shi, X., Wang, Z. and Sun, S. (2007). Antimicrobial activity of clove and rosemary essential oils alone and in combination. Phytotherapy Research 21(10): 989-994.
Gill, A. O. and Holley, R. A. (2006a). Disruption of E. coli, Listeria monocytogenes and Lactobacillus sakei cellular membranes by plant oil aromatics Applied Environmental Microbiology, 70: 5750-5755.
Gill, A. O. and Holley, R. A. (2006b). Inhibition of membrane bound ATPases of E. coli and Listeria monocytogenes by plant oil aromatics. International Journal of Food Microbiology, 111: 170-174.
Gupta, C., Garg, A. P., Uniyal, R. C. and Kumari, A. (2008). Antimicrobial acivity of some herba oila against common food-borne pathogens. African Journal of Microbiology, 2: 258-261.
Haripriyan, J., Omanakuttan, A., Menon, N. D., Vanuopadath, M., Nair, S. S., Corriden, R., Bipin, G. N., Nizet, V. and Geetha, B. K. (2018). Clove bud oil modulates pathogenicity phenotypes of the opportunistic human pathogen Pseudomonas aeruginosa. Scientific reports, 8: 3437.
Hemalatha, R., Nivetha, P., Mohanapriya, C., G., Sharmila, C., Muthukumaran, C. and Gopinath, M. (2016). Phytochemical composition, GC-MS analysis, In vitro antioxidant and antibacterial potential of clove flower bud (Eugenia Caryophyllus) methanolic extract. Journal of Food and Science Technology, 53(2): 1189-1198.
Ilechukwu, C. G. A., Ubesie, A. C., Ojinnaka, C. N., Emechebe, G. O. and Iloh2, K. K. (2014). Otitis Media in Children: Review Article. Open Journal of Pediatrics, 4: 47-53.
Kalaivani, G. and Vidhya, A. (2014). Watermelon seeds extracts and fresh juice for preliminary in vitro phytochemical screening and its antimicrobial activity analysed by TLC-bioautography technique. International Journal of Advancements in Life Sciences, 7: 74-80.
Kon K. V. and Rai M. K. (2012). Plant essential oils and their constituents in coping with multidrug-resistant bacteria. Expert Reviews of Anti-Infective Therapy 10(7):775-790.
Lee, Y. (2016). Cytotoxicity evaluation of essential oil and its components from Zingiber officinale Roscoe. Toxicological Research, 32(3): 225-230.
Leonardo G. F., Glaucius O. and Andricopulo A.D. (2018) From Medicinal Chemistry to Human Health: Current Approaches to Drug Discovery or Cancer and Neglected Tropical Diseases. An Acad Bras Cienc. https://dx.doi.org/10.1590/0001-3765201820170505.
Lopez – Vallejo F., Giulianotti M.A., Houghten R.A., Medina – Franco J. L. (2012). Expanding the medicinally relevant chemical space with compound libraries. Drug Discov Today. 17(13-14): 718 – 726.
Naveed, R., Hussain, I., Tawab, A., Tariq, M., Rahman, M., Hameed, S., Mahmood, M. S., Siddique, A. B. and Iqbal, M. (2013). Antimicrobial activity of the bioactive components of essential oils from Pakistani spices against Salmonella and other multi-drug resistant bacteria. Complementary and Altenative Medicine¸13: 265-275.
Obuotor, T. M., A.O. Kolawole, O. E. Apalowo and A.J. Akamo (2021). Metabolic profling, ADME pharmacokinetics, molecular docking studies and antibacterial potential of Phyllantus muellerianus leaves. Advances in Traditional Medicine https://doi.org/10.1007/s13596-021-00611-5.
Odenholt, I., Lowdin, E. and Cars, O. (2001). In-vitro pharmacodynamicsof telithromycin against respiratory tract pathogens. Antimicrobal Agents Chemotherapy, 45: 23-29.
Okore V. C. (2010). Principles of Pharmaceutical Microbiology. (2ndedn), Ephrata Publishers, Nigeria.
Omidbaigi, R., Yahyazadeh, M., Zare, R.and Taheri, H. (2007). The in vitro action of essential oils on Aspergillus flavus,” Journal of Essential Oil-Bearing Plants,10(1): 46–52.
Oni, A. A., Bakare, R. A., Nwaorgu, O. G. B., Ogunkunle, M. O. and Toki, R. A. (2001). Bacterial agents of discharging ears and antimicrobial sensitivity patterns in children in Ibadan, Nigeria. West African Journal Medicine, 20: 131.
Oviya, V. J., Vishnupriya, V. and Gayathri, R. (2016). Comparative analysis of antioxidant and free radicals scavenging activity of lemongrass oil and lavender oil. Asian Journal of Pharmaceutical and Clinical Research, 9, 136-139.
Oyedemi, S. O., Okoh, A. I., Mabinya, L. V., Pirochenva, G. and Afolayan, A. J. (2009). The proposed mechanism of bactericidal action of eugenol. Α-terpineol and γ-terpinene against Listeria monocytogenes, Sreptococcus pyogenes, Proteus vulgaris and Escherichia coli. African Journal of BiotechnologyI, 8(7): 1280-1286.
Pankey, A. and L.D. Sabath. (2004). Clinical relevance of bacteriostatic versus bactericidal mechanisms of action on the treatment of Gram-positive bacterial infections. Clinical and Infectious Diseases, 38(6): 864-870.
Peter. Y. Y. and Wong, D. D. (2006). Studies on the dual antioxidant and antibacterial properties of parsley (Petroselinum crispum) and cilantro (Coriandrum sativum) extracts. Food Chemistry 97:505–515.
Phyllis, B. and James, B. (2000). Prescription for Nutritional Healing, 3rd ed., Avery Publishing, p94.
Prakash, R., Juyul, D., Negi, V., Pal, S., Adekhandi, S., Sharma, M. and Sharma, N. (2013). Microbiology of chronic suppurative otitis media in tertiary care setup of Uttarakhand state. Indian Science Abstracts- International Journal of Medical Science, 5: 282-287.
Prashar, A., Locke, I. C and Evans, C. S. (2006). Cytotoxicity of Clove (Syzgium aromaticum) oil and its major components to human skin cells. Cell Proliferation, 39: 241-248.
Prieto, P., Pineda, M. and Aguilar, M. (1999). Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: Specific application to the determination of vitamin E. Analytical Biochemistry, 269: 337-341.
Qureishi, A., Lee, Y., Belfield, K., Birchall, J. P. and Daniel M. (2014). Update on otitis media- prevention and treatment. Infection and Drug Resistance, 7:15-24.
Rios J.L. and Recio M.C. (2005). Medicinal plants and antimicrobial activity. Journal of Ethnopharmacology 100:80-84.
Saeed, S. and Tariq, P. (2008). In Vitro Antibacterial activity of Clove against Gram negative bacteria. Pakistan Journal of Botany, 40(5): 2157-2160.
Sattar, A. A., Alamgir, A., Hussain, Z., Sarfraz, S. Nasir, J. and Badar, A. (2012). Bacterial Spectrum and their sensitivity pattern in patients of chronic suppurative otitis media. Journal of the college of physicians and surgeons Pakistan, 22: 128-129.
Sharma, K., Aggarwal, A. and Khurana, P. M. (2010). Comparison of bacteriology in bilaterally discharging eras in chronic suppurative otitis media. Indian Journal of Otolaryngol Head and Neck Surgery, 62: 153-157.
Sierra, A., Lopes, P., Zapata, M. A., Vanegas, B., Castrejon, M. M and DeAntonio, R. (2011). Non-typeable Haemophilus influenza and Streptococcus pneumonia as primary causes of acute otitis media in Colombian children: a prospective study. BioMed Central infectious diseases, 11(4):1–11.
Singh, G., Kapoor, I. S., Singh, P., de Heluani, C. S., de Lampasona, M. P., and Catalan, C. A. (2008). Chemistry, antioxidant and antimicrobial investigations on essential oil and oleoresins of Zingiber officinale. Food Chemistry Toxicology, 46(10):3295-302.
Singh, P. S. R., Prakash, B., Kumar, A., Singh, S., Mishra, P. K. and Dubey, N. K. (2010). Chemical profile, antifungal, antiaflatoxigenic and antioxidant activity of Citrus maxima and Citrus sinensis (L.) Osbeck essential oils and their cyclic monoterpene, DL-limonene. Food Chemistry and Toxicology, 48: 1734-1740.
Stojkovic, D. S., Zlukovic, J., Sokovic, M., Glamocija, J., Ferreira, C. F. R. Jankovic, T. and Maskimovic, Z. (2013). Antibacterial activity of Veronica montana L extract and of protocatechuic acd incorporated in s food system. Food and Chemical Toxicology, 55: 209-213.
Thosar, N., Basak, S., Bahadure, R. N. and Rajurkar, M. (2013). Antimicrobial efficacy of five essential oils against oral pathogens: An in vitro study. European Journal of Dentistry, 7: 71-77.
Tiwari, B. K., Valdramidis, V. P., O’ Donnell, C. P., Muthukumarappan, K., Bourke, P and Cullen, P. J. (2009). Applicaton of natural antimicrobials for food preservation. Journal of Agriculture and Food Chemistry, 67: 5987-6000.
Trongtokit, Y., Rongsriyam, Y., Komalamisra, N. and Apiwathnasorn, C. (2005). Comparative repellency of 38 essential oils against mosquito bites. Phytotherapy Research, 19(14): 303-309.
Walsh, S. E., Maillard, J. Y., Russell, A. D., Catrenich, C. E., Charbonneau, D. L. and Bartolo, R. G. (2003). Activity and mechanisms of action of selected biocidal agents on Gram-positive and Gram-negative bacteria. Journal of Applied Microbilogy, 94: 240-247.
Westh, H., Zinn, C. S., Rosdahl, V. T. and Sarisa Study Group. (2004). An international multicenter study of antimicrobial consumption and resistance in Staphylococcus aureus isolates from 15 hospitals in 14 countries. Microbial Drug Resistance, 10: 169-176.
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