H. N. K. S. Pathirana, S. H. M. P. Wimalasena, B. C. J. De Silva, S. Hossain, Heo Gang-Joon


The antibacterial activity of clove (Syzygium aromaticum) essential oil (CEO) and eugenol was tested against seven Gram-negative and nine Gram-positive fish pathogenic bacteria isolated from cultured olive flounder (Paralichthys olivaceus) in Korea. Eugenol was >99% concentrated and CEO consisted of seven chemical compounds including 83.63% of eugenol. Disk diffusion assay, MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration) tests showed CEO and eugenol inhibited the growth of both Gram-negative and Gram-positive bacteria. CEO (MBC/MIC= 1-2) and eugenol (MBC/MIC= 1-4) were bactericidal for all tested strains. The percentage of relative inhibition zone diameter exhibited high values at higher concentration of both CEO and eugenol. Positive correlation was observed between MAR index values and MIC values of CEO (r =0.61) and eugenol (r =0.15). Since antibacterial activity of CEO and eugenol were effective against fish pathogenic bacteria, the use of CEO and eugenol could be advantageous to treat bacterial infections in aquaculture.

Key words: antibacterial activity; clove essential oil (CEO); eugenol; fish pathogenic bacteria; olive flounder


Protibakterijsko delovanje eteriÄnega olja nageljnovih žbic (Syzygium aromaticum) (CEO) in evgenola je bilo testirano pri sedmih vrstah po Gramu negativnih in devetih vrstah po Gramu pozitivnih patogenih bakterij, ki so bile izolirane iz gojenega morskega lista (Paralichthys olivaceus) v Južni Koreji. Evgenol je bil 99-odstotno koncentriran, CEO pa je sestavljalo sedem kemiÄnih spojin, med njimi je bilo najveÄ evgenola, ki je predstavljal 83,odstotka eteriÄnega olja. Metoda difuzije v trdem gojiÅ¡Äu, minimalna zaviralna koncentracija (MIC) in minimalna baktericidna koncentracija (MBC) so pokazale, da sta CEO in evgenol zavirala rast tako po Gramu negativnih kot po Gramu pozitivnih bakterij. CEO (MBC/MIC = 1-2) in evgenol (MBC/MIC = 1-4) sta bila baktericidna za vse testirane seve. Odstotek velikosti relativne zaviralne cone je bil visok pri viÅ¡ji koncentraciji CEO in evgenola. Pozitivna soodvisnost je bila opažena med vrednostmi indeksa MAR in vrednostmi MIC pri CEO (r = 0,61) in evgenolu (r = 0,15). Ker sta CEO in evgenol pokazala uÄinkovito delovanje proti patogenim bakterijam rib, bi bila uporaba CEO in evgenola lahko uporabna za zdravljenje bakterijskih okužb v ribogojniÅ¡tvu.

KljuÄne besede: antibakterijsko delovanje; eteriÄno olje klinÄkov (CEO); evgenol; patogene bakterije rib; morski list

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(1) Jee BY, Shin KW, Lee DW, Kim YJ, Lee MK. Monitoring of the mortalities and medications in the inland farms of olive flounder. Paralichthys olivaceus, in South Korea. J Fish Pathol 2014; 27: 77–83.

(2) Kim SM, Jun LJ, Park MA, Jung SH, Jeong JB. Monitoring of emaciation disease in cultured olive flounder in Jeju (2010–2013), Korea. Korean J Fish Aquat Sci 2015; 48: 719–24.

(3) Nho SW, Shin GW, Park SB, et al. Phenotypic characteristics of Streptococcus iniae and Streptococcus parauberis isolated from olive flounder (Paralichthys olivaceus). FEMS Microbiol Lett 2009; 293: 20–7.

(4) Wei LS, Wee W. Chemical composition and antimicrobial activity of Cymbopogon nardus citronella essential oil against systemic bacteria of aquatic animals. Iranian j Microbiol 2013; 5: 147–52.

(5) Akbary P. In vitro inhibitory activity of the leaf methanol extract of green tea (Camellia sinensis) against Lactococcus garvieae and Aeromonas hydrophila isolated of rainbow trout (Oncorhynchus mykiss). Adv Microbiol 2014; 4: 829–34.

(6) Pongsak R, Parichat P. Protective effect of clove oil-supplemented fish diets on experimental Lactococcus garvieae infection in tilapia. Biosci Biotechnol Biochem 2009; 73: 2085–9.

(7) Burt S. Essential oils: their antibacterial properties and potential applications in foods: a review. Int J Food Microbiol 2004; 94: 223–53.

(8) Ayoola1 GA, Lawore1 FM, Adelowotan T, et al. Chemical analysis and antimicrobial activity of the essential oil of Syzigium aromaticum (clove). Afr J Microbiol Res 2008; 2: 162–6.

(9) Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils: a review. Food Chem Toxicol 2008; 46: 446–75.

(10) Huang X, Feng Y, Huang Y, Li H. Chemical composition, antioxidant and the possible use as skin-care ingredient of clove oil (Syzygium aromaticum (L.) Merr. & Perry) and citronellaoil (Cymbopogon goeringii) from China. J Essent Oil Res 2013; 25: 315–23.

(11) Sabahat S, Tariq P. In vitro antibacterial activity of clove against Gram negative bacteria. Pak J Bot 2008; 40: 2157–60.

(12) Oyedemi SO, Okoh AI, Mabinya LV, Pirochenva G, Afolayan AJ. The proposed mechanism of bactericidal action of eugenol, terpineol and γ-terpinene against Listeria monocytogenes, Streptococcus pyogenes, Proteus vulgaris and Escherichia coli. Afr J Biotechnol 2009; 8: 1280–6.

(13) Gill AO, Holley RA. Inhibition of membrane bound ATPases of Escherichia coli and Listeria monocytogenes by plant oil aromatics. Int J Food Microbiol 2006; 111: 170–4.

(14) Nazzaro F, Fratianni F, Martino LD, Coppola R, FeoVD. Effect of essential oils on pathogenic bacteria. Pharmaceuticals 2013; 6: 1451–74.

(15) Bennis S, Chami F, Chami N, Rhayour K, Tantaoui-Elaraki A, Remmal A. Eugenol induces damage of bacterial and fungal envelope. Moroccan J Biol 2004; 1: 33–9.

(16) Njau EA, Alcorn J, Ndakidemi P, Chirino-Trejo M, Buza J. Antimicrobial and antioxidant activity of crude extracts of Rauvolfia caffra var. caffra (Apocynaceae) from Tanzania. Int J Biol 2014; 6: 156–67.

(17) Hammer KA, Carson CF, Riley TV. Antimicrobial activity of essential oils and other plant extracts. J Appl Microbiol 1999; 86: 985–90.

(18) CLSI. Performance standards for antimicrobial susceptibility testing of bacteria isolated from aquatic animals: second informational supplement. CLSI document VET03/VET04-S2. Wayne, PA : Clinical and Laboratory Standards Institute, 2014.

(19) Seongwei L, Musa N, Wee W, Musa N. Chemical composition and antimicrobial activity of the essential oil of Syzygium aromaticum flower bud (clove) against fish systemic bacteria isolated from aquaculture sites. Front Agric China 2009; 3: 332–6.

(20) Cortés-Rojas DF, de Souza CRF, Oliveira WP. Clove (Syzygium aromaticum): a precious spice. Asian Pac J Trop Biomed 2014; 4: 90–6.

(21) Sohilait HJ. Chemical composition of the essential oils in Eugenia caryophylata, thunb from Amboina Island. Sci J Chem 2015; 3: 95–9.

(22) Chaieb K, Hajlaoui H, Zamantar T, et al. The chemical composition and biological activity of clove essential oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): a short review. Phytother Res 2007; 21: 501–6

(23) Rehab MA, Zeinab SH. Eugenol and linalool: comparison of their antibacterial and antifungal activities. Afr J Microbiol Res 2016; 10: 1860–72.

(24) Bharath MR, Azeem MA, Keerthan HV. Antimicrobial activity of clove extracts against food borne pathogens Escherichia.coli, Salmonella.tyhimurium, Staphylococcus aureus and Listeria monocytogenes and GC-MS analysis of extracts. Int J Pharm Bio Sci 2017; 8: 321–9.

(25) Fagere ZO, Zoheir A, Magboul A. Antibacterial activity of clove oil against some microorganisms at Khartoum State. Adv Med Plant Res 2016; 4: 122–8.

(26) Shahavi MH, Hosseini M, Jahanshahi M, Meyer RL, Darzi GN. Clove oil nanoemulsion as an effective antibacterial agent: Taguchi optimization method. Desalin Water Treat 2015; 57: 18379–90.

(27) Devi KP, Nisha SA, Sakthivel R, Pandian SK. Eugenol (an essential oil of clove) acts as an antibacterial agent against Salmonella typhi by disrupting the cellular membrane. J Ethnopharmacol 2010; 130: 107–15.

(28) Chitanand MP, Kadam TA, Gyananath G, Totewad ND, Balhal DK. Multiple antibiotic resistance indexing of coliforms to identify high risk contamination sites in aquatic environment. Indian J Microbiol 2010; 50: 216–20.



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