Evaluation of Combined Antibacterial Effects of Eugenol, Cinnamaldehyde, Thymol, and Carvacrol Against E. Coli with an Improved Method

See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/38072806

Evaluation of Combined Antibacterial Effects of Eugenol, Cinnamaldehyde, Thymol, and Carvacrol against E. coli with an Improved Method

ARTICLE in JOURNAL OF FOOD SCIENCE · SEPTEMBER 2009 Impact Factor: 1.7 · DOI: 10.1111/j.1750-3841.2009.01287.x · Source: PubMed

CITATIONS READS 82 443

4 AUTHORS, INCLUDING:

Ruisong Pei Bao-Ping Ji University of Wisconsin–Madison China Agricultural University

12 PUBLICATIONS 232 CITATIONS 18 PUBLICATIONS 244 CITATIONS

SEE PROFILE SEE PROFILE

All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Ruisong Pei letting you access and read them immediately. Retrieved on: 20 February 2016 T R against Carvacrol and Thymol, Cinnamaldehyde, Eugenol, of Effects Antibacterial Combined of Evaluation JFS ute erdcinwtotpriso sprohibited is permission without reproduction Further in- Direct China. Beijing, [email protected]). District, (E-mail: Zhou Agricultural Haidian author China to Donglu, quiries Engineering, Qinghua Nutritional and 17 Univ., Science Food 6/17/2009 of Accepted College 2/19/2009, Submitted 20090150 MS ors19) o ntne twudne oeta wfl more twofold than more need would and it Smid instance, 1983; For (Shelef 1999). would situations Gorris effects particular antimicrobial some the in because weakened achieve be foods, to in needed effect is same addition the these greater of even taste the besides, by affected components; be could foods the However, of foods quality activities. selected organoleptic antimicrobial of and life to shelf safety possible the their is extend considering to it Therefore, preservatives as 2004). risk them (Burt no use customers present of to health considered the are to by and registered Commission been European have the more eugenol, carvacrol, and including and safe components, thymol, their cinnamaldehyde, as and regarded EOs are Many which nutritional. products, food processed of growth on and thymol cinnamaldehyde, carvacrol, of re- effects researchers antimicrobial the Other ported 2004). others and (Bagamboula investigated against effective were thyme and of oregano strains the of oils essential the o:10.1111/j.1750-3841.2009.01287.x doi: n tes20) h niioyefcso hm n ai O,car- EOs, toward thymol basil and and vacrol, thyme of effects inhibitory The 2003). others and against of effective oils are thymol essential clove the isomer and that thyme its shown have and Studies 2005). carvacrol others are and (Burt oils these of components rial nIpoe Method Improved an C UI 09Isiueo odTechnologists Food of Institute 2009 nicesn ubro epepee aua n minimal and natural prefer people of number increasing An BTAT nti td,teatbceilatvte fegnl inmleye hml avco,adtheir and carvacrol, thymol, cinnamaldehyde, eugenol, of activities antibacterial the study, against this combinations In ABSTRACT: hs opnnscudb iiie,mkn tpsil oadte ofosa rsraie.I diin this addition, effects. In combined preservatives. evaluate as to of foods way comprehensive smell to and them unpleasant accurate add of more to a impacts possible provided negative it method the making evaluation Consequently, improved minimized, respectively. be mg/L, could carvacrol and 100 components thymol, 4 treatments and cinnamaldehyde, these respectively; eugenol, 100, of mg/L, 100, MICs 400 400, combination, jointly. of to and means (EC) decreased ef- By 400, indices. Combination synergistic 400, 2 revealed the the 1600, to thymol/carvacrol of were according and fects components Effect carvacrol/eugenol, 4 and thymol/eugenol, the (FIC) cinnamaldehyde/eugenol, of concentration with MICs inhibitory that fractional indicated per- of results was which indices method, testing The improved the combination an with the on evaluated Second, were based effects components. was combined 4 the Finally, the method. of chequerboard using (MIC) formed concentration inhibitory minimal the mine - et aebe xmndrcnl.I a enrpre that reported been has It recently. examined been compo- their have and nents (EOs) oils essential of effects antibacterial he SONG ewrs niatra activity, antibacterial Keywords: :Fo irbooyadSafety and Microbiology Food M: .coli E. P EI ,F Dra n en 00.Temjrantibacte- major The 2000). Deans and (Dorman ENG ailscereus Bacillus Z HOU Introduction hglasonnei Shigella .coli E. ,B AO - Vlr n ie 2005). Giner and (Valero eeivsiae eaaey is,bohmcodlto sa a dpe odeter- to adopted was assay macro-dilution broth First, separately. investigated were PING R itramonocytogenes Listeria and J I , .coli E. AND hglaflexneri Shigella J . seta iscmoet vlainmto,synergism method, evaluation component, oils essential , ING uhr r with are Authors X U (Singh were ie onl odvlpamr opeesv a oqatf the combination. quantify EOs to of way effects comprehensive synergistic more in- a 2 develop these to use to jointly tried dices we reports, previous the on based Therefore, against acids components our organic typhimurium of In and chelator effects 2008). with combined others or the and themselves tested Si we and 2005; (Shin studies, others EOs previous and of Filoche abilities 2003; antimicrobial the Kang study used to also method have researchers this Many medicine. antimicro- in drugs the of test abilities to bial used commonly is index (FIC) concentration combination. EOs of effects evalu- synergistic to the way standardized quantify and and effective ate an is develop there to Thus, need 2002). clear others a and Delaquis 1998; 2000; others others and and (Fyfe re- Ettayebi outcomes the nonstandard evaluate to to leads which re- indices sults, various these adopt of and results testing the the perform combination to compare methods different to choose researchers hard because ports is it However, oth- and 2008). (Solomakos ester ers Holley acid fatty and diglycerol and (Blaszyk nisin monolaurin and 1998), and citrate sodium 2007), and others (Chouliara atmosphere stud- would modified previous with tymol in combined and reported including ies, also nisin were of methods Other concentration decreased. required be the bacte- that pathogenic so foodborne ria, the control and effectively nisin of could combination thymol the that found (2000) others and of Ettayebi viability the with reduce thymol could treatment and heat cinnamaldehyde, mild carvacrol, of the effects that reported combined (2000) others and Karatzas treatments. and of agents variety antibacterial a other with combination in used been have 1994). Shelef tenfold and (Pandit and sausage 2001) liver others pork in and more (Karatzas milk semiskimmed in EOs h hqebadmto obndwt rcinlinhibitory fractional with combined method chequerboard The components EOs the activities, antimicrobial the enhance To o.7,N.7 2009 7, Nr. 74, Vol. sn h Cidx(huadohr 07,2007b). 2007a, others and (Zhou index EC the using .coli E. — ORA FFO SCIENCE FOOD OF JOURNAL with .monocytogenes L. Salmonella M 379 . M: Food Microbiology & Safety 1) − < E. coli 0.1 (log DP < II 0.01) confidence limit. Dif- 90%): linked with the defini- ≤ Log DP > P − I are shown in Figure 1. The log DP of Log DP represented the Log DP of the combina- coli II . = E 0.05) or 99% ( 3.85 were found in groups treated with 0, 400, EC − ≤ Results and Discussion P and Log DP I 0.05) compared with control tube. Therefore, according to ≤ Two kinds of indices were adopted toThe evaluate FIC, the which interactions. is the sum of the FICs for each component; and The EC, which was described in our previous studies (Zhou and The ANOVA was performed using a mixed-model procedure with The antibacterial activities of eugenol, cinnamaldehyde, thymol, The synergistic effects of the combinations were determined From the results of the antibacterial activity of each compo- Only when the combinations showed all these 3 principles that (1) The decrease in populations ( (2) When there was significant difference (analyzed by the one- (3) Synergy was defined as a 2-log decrease of population in the P suspensions and aliquots of 1 mLtration of of each 4 component times at the ain concen- targeted the final tubes. One concentration tube were with dispensed ponents 4 was mL set of as MHB control. without The addingthrice. experiment any The was com- MICs carried were out determined at as least described previously. the FIC is calculated as follows: forMIC component of A component the A FIC in equals combination the dividednent by the A MIC alone. of The compo- interactions areindex considered is synergistic less if than the 1, additive FIC ifFIC the index FIC is is equal between to 1, 1greater indifferent if than and the 2 2, (Franzot and and Casadevall antagonistic 1997). if the FIC indexothers is 2007a) and the EC is calculated using the following formula: Statistical analysis of data Antibacterial activity of each component a randomized complete block design with repeatedThe measurements. experiment was repeated at leastconverted into 3 logarithm times. values Colony to counts determine the were ences significant at the differ- 95% ( and carvacrol against ferences among the treatments were examinedcant by differences the test (SAS least version signifi- 9.0, SAS Inst., Cary, N.C., U.S.A.). tion system and the single agent, respectively. based on 3 principles. tion of DP,it was concluded that only when DP The Log DP 2.92, 2.03, 0.38, and the definition of MIC described previously, the1600 MIC of mg/L; eugenol also, the was MBC ofMICs eugenol of was cinnamaldehyde, 1600 thymol, and mg/L. carvacrol Similarly, the were400 400, mg/L, 400, respectively; and the MBCs of eugenol, cinnamaldehyde,mol, thy- and carvacrol were 1600, 1600, 800, and(data 800 are mg/L, not respectively totally shown in Figure 1). nent, cinnamaldehyde, thymol, and carvacrolantibacterial revealed effects. the This similar was in line with the results of Helander that the combinations of variousterial reagents activity. had significant antibac- way analysis of variancetivity [ANOVA]) of between the the combination antibacterialmeant and the ac- the combination was individuals, more respectively, effective. which drug combination group compared withagent at the the most end effective of 24 single they h. were regarded to have a synergistic effect. 800, and 1600 mg/Lor of eugenol. more Samples of treated with eugenol( 1600 significantly mg/L reduced the number of C ◦ 0). The ≤ is the initial C. ) ◦ m-pore filter; 0 0 N μ N 98%) and thymol ≥ (Log 98%) were obtained ; furthermore, an im- − ≥ ) N E. coli Vol. 74, Nr. 7, 2009 CFU/mL. All tubes were in- — 5 (Log 10 = ) × 0 N / N 98%) and eugenol ( Log ( ≥ = C. Cells were prepared by 16-h culture in Mueller Materials and Methods ◦ C for 24 h, 1 mL portion was removed from each ◦ was tested for susceptibility to the 4 components by 3). Log DP CGMCC 1.487 (obtained from China General Microbio- ≤− JOURNAL OF FOOD SCIENCE CFU/mL approximately. The number of cells in the suspen- E. coli 6 10 MIC is defined based on the logarithmic difference in population Combination testing was performed by the chequerboard The Cinnamaldehyde ( E. coli Eugenol, cinnamaldehyde, thymol, and carvacrol were 4 com- A 16-h culture was diluted with MHB to achieve an inoculum of is the population after incubation for 24 h and 99%) were obtained from Sigma (St. Louis, Mo., U.S.A.). Certain 380 × ≥ amount of each component filter-sterilized (0.22- ( Millipore Corp, Bedford, Mass., U.S.A.)tain was a stock added solution to of 3200 MHB mg/L to with ob- Tween-80 as emulsifier. from Fluka (Buchs, Switzerland). Carvacrol ( tube for colony counting by decimal dilution inchloride 0.85% (w/v) solution, sodium and platedcarried out out onto at MHA least thrice. The experiment was (Log DP). The Log DP isand expressed Lin by 1995): the following equation (Fang M Combination testing Antimicrobial susceptibility testing EO components Bacterial strains and growing conditions An improved evaluation method of combination effects . . . cubated at 37 sions was estimated by duplicatetion plating on from MHA and tenfold counting serial the dilu- colonies after incubation at 37 method as previously described (Franzot and Casadevallrial 1997). twofold Se- dilutions of eachpared component at stock solution 4 were times pre- from the half to strength the of one-eighth the of the final MIC. concentration A ranging total of 2 mL of the cell broth macrodilution assay (Wilson and othersdilutions 2005). Serial of twofold the 4centrations components ranging as were 3200, prepared 1600, ineach 800, component. MHB 400, One 200, with tube and con- with 100 theas mg/L same control. for volume Inocula of were MHB added was intotial set all inoculum the of tubes approximate to 5 achieve an ini- logical Culture Collection Center, Beijing,on China) Mueller–Hinton was agar maintained (MHA)China) (Beijing slants at Chemical 4 Corp., Beijing, monly usedcinnamon, antibacterial oregano, and thyme components (Demo andothers others 2001; of 2003; Kwon and Valero the andstudy Giner was EOs 2005). to The investigate of major theactivities of separate objective these 4 clove, and of components combined against this antibacterial proved method was developed to evaluate andcombinations. quantify synergistic –Hinton broth (MHB) (Beijing Chemical Corp.) at 37 1 population. MIC is defined as themaintenance lowest or concentration reduction resulting of in inoculums viability (Log DP minimum bactericidal concentration (MBC) is definedcentration as where the 99.9% con- or more(Log of DP the initial inoculums are killed N for 24 h. M: Food Microbiology & Safety al,tecmiain f() 3,() 6,() 8,ad()la to lead (9) and (8), (7), (6), (4), (3), (2), of Particu- combinations effects. synergistic the larly, had underlines) by (marked binations of growth the Table2.Accordingtothe3principlesestablishedearlier,9com- on systems combination 24 ueo/hmlwr togr(FIC stronger eugenol/cinnamaldehyde, were of effects eugenol/thymol synergistic the carvacrol which and among cinnamaldehyde ad- between only found (FIC The was 1. Table effect in ditive listed were tested substances timicrobial aintm,adtmeauewudas fetteotoe the outcomes the affect incu- also pH, experiments. would phase, temperature as growth and such time, inoculums, factors bation of some volume Besides, medium, activity. culture antimicrobial perform of to chosen tests were the that diffusion, methods like bioautographic methods or different dilution, the these as from explained derived be order might reports antimicrobial of discrepancy The 2007). vacrol or- carvacrol antimicrobial was the der that 1998). reported others others and and Friedman (Helander However, carvacrol and thymol that namaldehyde, found who others and . . . effects combination of method evaluation improved An niatra fet fcmiaingroups combination of effects Antibacterial ilsbtne ntegot of antimicro- growth combined the with on treatments substances bial of Effect --- 1 Table .coli against order E. antimicrobial the found others and Michiels while say 37 avco ueo 0 6010800.75 0.50 0.75 800 400 100 100 100 200 1600 1600 400 400 0.50 400 1.00 0.75 400 400 Eugenol Eugenol 200 100 100 Carvacrol 200 200 1600 400 400 400 Carvacrol Thymol 400 400 Eugenol Thymol Carvacrol Cinnamaldehyde Thymol Cinnamaldehyde Cinnamaldehyde ABABABFIC h I nie agn rm05 o1frtecmie an- combined the for 1 to 0.50 from ranging indices FIC The ◦ codn oteFCindex. FIC the to according C = > nsalitsia iuain a cinnamaldehyde was simulations intestinal small in ) h etcmiain eeldsnritc(FIC synergistic revealed combinations rest The 1). Components hml(remnadohr 02 ihesadothers and Michiels 2002; others and (Friedman thymol > cinnamaldehyde E . coli ln MIC Alone (mg/L) a niie qal ycin- by equally inhibited was > ≤ .coli E. hmli irpaeas- microplate a in thymol .) h fet fthe of effects The 0.5). E . nMBsoe at stored MHB in coli I (mg/L) MIC Combined eelse in listed were ≤ ≥ 0.75), car- et demonstrated nents plant of against enteriditis acid effects Salmonella benzoic combined of regarded derivatives and others con- oils and showed Fyfe isobologram co- the shape. cilantro, when cave dill, (2002) EOs of others eucalyptus fractions and and mixed riander, Delaquis of Besides, effects 2007a, synergistic others 2008). and defined others Zhou and 2005; Si others been (Shin 2007b; and have effects Filoche EC combined 2003; evaluate and Kang to and FIC researchers of many indices by Two adopted EOs. of effects synergistic but index. index FIC EC the the by to additive according synergistic was mg/L) mg/L) (200 (200 on cinnamaldehyde carvacrol of based group the Similarly, effect index. synergistic EC the showed thymol and cinnamaldehyde in- FIC the cin- to (FIC according dex between 2 synergistic interaction was the the thymol on instance, and eval- based For namaldehyde 2 cases. conclusions the some different in of got indices methods also we mathematical indices, different uation 2 addition, In the obvious. of not were because components other with effects tion hwdsrn niatra fetagainst effect cinnamaldehyde hand, antibacterial other strong the showed On components. weak- 4 the the was among alone est eugenol thy- of effect and antibacterial the cinnamaldehyde though with mol, combined when effect synergistic yegsi fet on effects synergistic the cin- showed for cinnamaldehyde/carvacrol except and combinations paired namaldehyde/thymol the all First, conclusions. tent the all kill DP could and effect bactericidal a fcmiaintsigadtemteaia rcse,i could it processes, mathematical the and results of testing comparison combination the of From syn- components. the EO determine of to effects method ergistic precise and comprehensive more a reports. the of results combination, the EOs compare to of impossible to effects it way making synergistic standardized the and quantify effective others and an evaluate and not (Fyfe is alone there used However, components 1998). the of effects inhibitory urnl,teeaesvrldfeetmtost eemn the determine to methods different several are there Currently, hog oprn h eut,w ol bansvrlconsis- several obtain could we results, the comparing Through een eitgae ao omnyue nie odevelop to indices used commonly major 2 integrated we Herein, =−∞ o.7,N.7 2009 7, Nr. 74, Vol. = ). .5,btnn fte4cmiaingop between groups combination 4 the of none but 0.75), 37 ( different signiﬁcantly different are with letters group same the within “ liter. per milligrams are concentration components’ the hne of cumulative changes on (car.) carvacrol and (thy.), thymol (eug.), (cin.), eugenol cinnamaldehyde of --- Effect 1 Figure P ≥ ≤ E ssnritcwe h obndcompo- combined the when synergistic as log ◦ o 4h “ h. 24 for C . 0.05). coli 10 — rae niiinta h u fthe of sum the than inhibition greater ORA FFO SCIENCE FOOD OF JOURNAL eod ueo eeldstronger revealed eugenol Second, . E . coli ‡ ” ouainsoe at stored population = E .coli E. . † .monocytogenes L. l h nt of units the All ” coli = ouain (Log populations hl t interac- its while Means M and 381 +

M: Food Microbiology & Safety +∞ +∞ +∞ +∞ +∞ +∞ +∞ 5.40 4.37 EC C according +∞ +∞ +∞ +∞ +∞ +∞ +∞ 5.03 1.470.82 2.61 4.28 1.73 2.45 2.48 1.84 2.95 ◦ was found in this . ∗∗ ∗∗ ∗ ∗ ∗∗ ∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗∗ ∗ ∗∗ E. coli +∞ 0.0001 0.0001 0.0128 00.05020.09320.9308 0.990.0115 1.89 0.79 2.25 0.9253 0.96 0.0001 0.01 0.0109 0.0001 0.800.0024 0.0001 0.05 0.0022 0.21060.0001 0.0286 1.480.0001 0.59 F > P ∗∗ ∗∗ ∗ ∗ ∗ ∗ ∗∗ ∗ ∗∗ ∗ ∗∗ ∗ ∗∗ ∗∗ ∗ ∗∗ ∗ ∗∗ in MHB stored at 37 Conclusions 0.0001 0.0003 0.0001 0.0001 0.0005 0.0001 0.0001 0.0001 0.0001 coli . E , comparatively, and the EC was b −∞ 0.10 0.0491 4.14 2.27 0.44 0.0205 0.95 0.0295 0.58 0.0118 −∞ −∞ −∞ −∞ −∞ −∞ −∞ − − − − − − ynergistic effectseugenol, were found thymol/eugenol, between carvacrol/eugenol, cinnamaldehyde/ and thy- Many researchers studied the combined effects of these 4 com- study. We supposed that themight mechanisms be of different, antimicrobial or action thedifferent combined bacterium. action might vary along with ponents. (Ettayebi andOlasupo others and others 2000; 2003). Karatzas However,combined systematic effects and studies about between others the theported. 2000; 4 In components this have4 not study, components been paired were re- combinationnamaldehyde/eugenol, determined in effects thymol/eugenol, detail. carvacrol/eugenol, betweenthymol/carvacrol Treatments the and showed with synergistic cin- effects. Italthough is noteworthy the that antibacterialpared ability with the of other eugenol 3tic components, effect. was it The revealed weaker MIC the400 of com- best synergis- eugenol mg/L by was combining decreased itvacrol. with from In cinnamaldehyde, summary, 1600 thymol, the and mg/L antibacterial car- abilities to were of enhanced the through 4 combination; components the lowest additiondecreased could significantly. be Consequently, the negativepleasant smell effects were minimized. of un- that eugenol and cinnamaldehyde acted onzymes. diverse Different proteins or researchers en- reported various combinationbetween effects carvacrol and thymoland others (Didry 2001). Synergistic and effect others against 1993; Lambert S mol/carvacrol. Through combination, the effectiveof the concentration 4 components could beAs significantly a lower result, their than impacts using on alone. thebe organoleptic quality of minimized. foods would ItshouldbeevaluatedbytheindicesofFICandECjointly.However, was recommended thatit the was necessary combined to point effects out24 that this h study period, was carried atmedium, out during 1 whereas pH the food (physiological system pH) is and more only complicated. in Further the culture 0 0 0 800 800 00 800 800 200 100 , making it easier E. coli Vol. 74, Nr. 7, 2009 — 00 0 0 800 800 0 200 Components (mg/L) populations were killed by one combination, the Log DP was E. coli a 0.01. 0 100 100 0 2.40 0.6349 0.5616 0.25 0.31 000 10000 200 200 100 0 0 0 0 0 200 400 100 0.67 400 400 0.0253 400 0.91 1.40 0.0596 1.34 0.9500 0.0477 0.2459 0.5914 1.24 0.3310 0.13 1.12 0.76 0.64 0.70 200 100 0 0 100100 200 100 0 0 0 0 1.05 2.15 0.0306 0.0305 100 0 0 400 1.21 0.0394 200200100 200100200 0 0 0 0 0 100 0 200 100 0 0 0 0 400 1.02 1.72 2.14 0.7440 0.1230 0.0348 0.1216 0.6081 0.13 1.22 1.08 0.45 ≤ When all the P ∗∗ Cinnamaldehyde Thymol Carvacrol Eugenol Log DP A B A B JOURNAL OF FOOD SCIENCE +∞ = 0.05; , ≤ The mechanism of antimicrobial action of EOs has been of inter- 382 −∞ Combination marked by underlines showed synergistic effects according to the EC index. P (3)(4) 100 0(7) 0 200 100 (1)(2) 200 200 (6) 0 200 to the EC index. (8)(9) 0 0 0 0 200 100 a b ∗ M (5) 0 200 Table 2 --- Effect of treatments with combined antimicrobial substances on An improved evaluation method of combination effects . . . be seen that, first,fects using the (additive, FIC synergistic, index, indifferent,judged all or possible between antagonistic) combined the ef- couldcould components. be be While decided only1st according synergistic method, to effects FIC the wasgistic figured EC effect. out index. to While evaluate Second, theLog the EC with DP degree index reflected the of how was syner- strongwas, based the the results on of combined ANOVA 3 incarnated antimicrobial the principles.gle activity relationship between The component sin- and thethe combination degree group, of and synergisticnation the effect. EC testing Therefore, showed should theto be results acquire determined of the combi- by all-sidedaction the results. could Consequently, 2 be all indicescombined classified. types group together and Besides, of the degree the inter- of synergisticponents antimicrobial effect could between activity com- be of reflected;sion furthermore, could more be veracious gained conclu- bymethods. comparing In the summary, by results comparing derived theit from results could the of be 2 the seen 2 that methods, thethymol/eugenol, interactions of carvacrol/eugenol, cinnamaldehyde/eugenol, andsynergistic. thymol/carvacrol were est to food microbiologists. Wendakoonpothesized and that the Sakaguchi hydroxyl (1993) group on hy- eugenol mightproteins, combine with preventing enzyme actioncinnamaldehyde while might the adhere to carbonyl proteins group toamino prevent on acid the decarboxylases. action Carvacrol of and thymolbic were hydropho- and prone tobacteria, disturb releasing lipopolysaccharides, the and outer increasing theability membrane perme- of of the Gram-negative cytoplasmic membrane1998; to Lambert ATP and (Helander others and 2001). others Basedhypothesized on that the the previous studies, synergistic we effectseugenol/thymol of might eugenol/carvacrol and be engenderedmol by disintegrated that the carvacrol outer and membrane of thy- for eugenol to enter the cytoplasmsynergistic and effects combine of with eugenol/cinnamaldehyde proteins; might the be caused by M: Food Microbiology & Safety huir ,KrtpnsA avii N otmnsM.20.Cmie fetof effect Combined 2007. MG. Kontominas IN, Savvaidis A, Karatapanis E, es- Chouliara of activity in Increase 2005. EJA. Veldhuizen HP, Haagsman R, Vlielander S, Burt in applications potential and properties antibacterial their oils: Essential 2004. S. Burt on citrate sodium and eugenol monolaurin, of Interaction 1998. RA. Holley M, Blaszyk basil and thyme of effect Inhibitory 2004. J. Debevere M, Uyttendaele CF, Bagamboula Engineer Senior and study. this Bi instructing Wangfu for Professor Liu Zonglin thank to like would We points. mentioned previously the of each consider will studies . . . effects combination of method evaluation improved An rno P aaealA 97 nuoadnL73 7 nacsteatvte of activities the enhances 872 L-743, Pneumocandin com- 1997. in A. oils Casadevall essential SP, of Franzot effects Antimicrobial 2005. CH. Sissons K, Soma SK, Filoche agT,LnL.19.Cmaaiegot of growth Comparative 1995. LW. Lin TJ, Fang thymol and nisin of effects Synergistic 2000. B. Rossi-Hassani JE, Yamani K, Ettayebi activity antibacterial plants: from agents and Antimicrobial 2000. carvacrol SG. Deans thymol, HJD, of Dorman activity Antimicrobial 1993. M. Pinkas L, Dubreuil activ- antibacterial N, the Didry of Determination 2001. J. Zigadlo B, Ramos M, Oliva M, Demo individual of activity Antimicrobial 2002. G. Mazza B, Girard K, Stanich PJ, Delaquis yeL rsrn ,SeatJ 98 niiinof Inhibition 1998. J. Stewart F, Armstrong L, Fyfe essen- plant of activities Bactericidal 2002. RE. Mandrell PR, Henika M, Friedman sonnei rs hce ratma,soe t4 at stored of meat, extension breast shelf-life chicken on fresh packaging atmosphere modified and oil essential oregano 68:919–26. Prot Food J stabilizers. against food thymol of addition and carvacrol components oil sential 94:223–53. Microbiol Food J Int review. a foods: Microbiol Food J Int organisms. pathogenic and 39:175–83. spoilage meat common of growth and linalool estragol, thymol, carvacrol, oils, essential mhtrcnB lcnzl against fluconazole B, amphotericin 20:221–5. Immunol Microbiol Oral digluconate. chlorhexidine with bination 33:391–403. Soc fragi domonas irbo et183:191–5. Lett Microbiol in activities antimicrobial on 88:308–16. Microbiol Appl J oils. volatile plant of 48:301–4. Pharmazie combination. in or alone cinnamaldehyde 85:87–90. Aliment Hig oils. essential of components pure the of J ity Int oils. essential eucalyptus and coriander 74:101–9. cilantro, Microbiol dill, Food of fractions mixed and ilol n oeo hi sltdcnttet against constituents isolated their 65:1545–60. of coli some Escherichia and oils tial 41:331–6. Chemother Agents cd h eeomn fsnritcatmcoilcmiain.ItJAntimicrob J Int combinations. 9:195–9. antimicrobial Agents synergistic of development the acid: enteriditis Salmonella and .flexneri S. namdfe topeensnmdlsse.JCi gi Chem Agric Chin J system. model atmosphere/nisin modified a in , itramonocytogenes Listeria odMcoil21:33–42. Microbiol Food . ycmiain fpatol n eiaie fbenzoic of derivatives and oils plant of combinations by Acknowledgments itramonocytogenes Listeria References rpoocsnoomn nvitro in neoformans Cryptococcus ◦ .Fo irbo 24:607–17. Microbiol Food C. ,and itramonocytogenes Listeria amnlaenterica Salmonella itramonocytogenes Listeria shrci coli Escherichia and p cmn towards -cymene ailssubtilis Bacillus aplbce jejuni Campylobacter odProt Food J . Antimicrob . 17 7by H7 O157: and Shigella .FEMS Pseu- and , aeoM ie J 05 fet fatmcoilcmoet fesnilol on oils essential of components antimicrobial of Effects 2005. MJ. Giner M, Valero edko N aauh .19.Cmie feto oimclrd n lv on clove and chloride sodium of effect Combined 1993. M. Sakaguchi CN, Wendakoon ooao ,GvrsA odsP osgo .20.Teatmcoilefc of effect antimicrobial The 2008. N. Botsoglou P, Koidis A, Govaris N, Solomakos Rahman In: preservation. food for antimicrobials Natural 1999. LGM. Gorris EJ, Smid and S-carvone of action Combined 2000. EP. Kets EJ, Smid MH, Bennik AK, Gorris Karatzas EJ, Smid I, Pol T, Mattila-Sandholm K, Latva-Kala HL, Alakomi IM, Helander lsp A izeadD,Gso J abdA 03 ciiyo aua antimi- natural of Activity 2003. A. Narbad MJ, Gasson DJ, Fitzgerald NA, Olasupo 2007. N. Dierick S, Smet De D, Fremaut J, Missotten minimum J, the Michiels of study A 2001. G-JE. Nychas P, Coote separation PN, cell Skandamis of RJW, inhibition Lambert and effects Bactericidal 2003. HD. Park CB, Yu JA, Kwon carvacrol of action combined The 2001. MHJ. Bennik EJ, Smid EPW, Kets AK, Karatzas WilsonB,AbrahamG,ManjuVS,MathewM,VimalaB,SundaresanS,NambisanB. huF iB hn ,JagH agZ iJ iJ a .20b h niatra effect antibacterial The 2007b. W. Yan J, Li J, Li Z, Yang H, Jiang H, Zhang B, thy- Ji of F, effect Zhou Synergistic 2007a. W. Yan J, Li J, Li Z, Yang H, Jiang H, Zhang B, Ji F, Zhou ig ,SnhR,Bui K ig .20.Efcc fpatesnilol santimi- as oils essential plant of Efficacy 2003. N. Singh AK, Bhunia RK, Singh A, Singh in and alone oil essential oregano of of effect Antibacterial oil 2008. Z. essential Zeng Z, the Liu J, of Hu H, activity Si Antifungal 2003. CA. 6:29–44. Safety Kang Food J S, spices. Shin of effects Antimicrobial 1983. LA. Shelef of Sensitivity 1994. LA. Shelef VA, Pandit rwhof growth 25:120–7. Microbiol Food storage. refrigerated during beef minced in rwhadboei mn omto of formation amine biogenic and growth 106:90–4. Microbiol Food J Int hm seta i,nsn n hi obnto against combination their and nisin, oil, essential thyme Dekker Marcel York: New preservation. food 285–308. of p Inc. Handbook editors. M, Dekker MS, 36:787–94. on treatment heat oil mild essential 46:3590–5. selected Chem Food of Agric action J the bacteria. Gram-negative of on Characterization components 1998. A. Von-Wright LGM, rba opud against compounds crobial 109:157– Sci Livest flora. gut pig com- the 60. against of activity interaction antimicrobial and the trans-cinnamaldehyde for binations and eugenol thymol, carvacrol, of and thymol oil, essential 91:453–62. oregano Microbiol of Appl action J carvacrol. of mode and concentration inhibitory on aldehyde cinnamic of 90:463–9. on pressure hydrostatic high and 05 niirba ciiyof activity Antimicrobial 2005. 56:410–3. Prot Food J extract. cle fcnaadhd,tyo,crarladtercmiain gis h food- the against combinations their and pathogen borne carvacrol thymol, cinnamaldehyde, of against acids organic and chelators typhimurium with combined carvacrol and mol 99:147–51. Ethnophar J rba gnsagainst agents crobial coli extended-spectrum Escherichia against antibiotics with combination 36:111–5. Microbiol Appl Lett ketoconazole. with synergism its and Kuntze officinalis inus phimurium o.7,N.7 2009 7, Nr. 74, Vol. ailscereus Bacillus etAp irbo 36:448–51. Microbiol Appl Lett . odPo 70:1704–9. Prot Food J . ..Fo irbo 11:57–63. Microbiol Food L.). ESImnlMdMcoil53:190–4. Microbiol Med Immunol FEMS . amnlatyphimurium Salmonella itramonocytogenes Listeria itramnctgnsscott monocytogenes Listeria ailscereus Bacillus NAL14i n h esr ulte fcro broth. carrot of qualities sensory the and in L2104 INRA shrci coli Escherichia ucm zedoaria Curcuma — itramonocytogenes Listeria ORA FFO SCIENCE FOOD OF JOURNAL itramonocytogenes Listeria etAp irbo 37:61–5. Microbiol Appl Lett . neoatraerogenes Enterobacter odSft 27:124–33. Safety Food J . and nhtos ees isTechnol Wiss Lebensm hotdogs. in and amnlaetrc eoa Ty- serovar enterica Salmonella .JAp irbo 89:296–301. Microbiol Appl J A. ucm malabarica Curcuma ct .JAp Microbiol Appl J A. Scott β itramonocytogenes Listeria nvitro In -lactamase-producing orsmr ( rosemary to nmcee mus- mackerel in gsah rugosa Agastache dose–response Salmonella Rosmar- tubers. M 383

M: Food Microbiology & Safety