Effects of Oregano Extract on the Inhibition of Selected Pathogens in Raw Beef Meat

Transcription

1 JOURNAL OF NUTRITION FASTING AND HEALTH Effets of Oregno Extrt on the Inhiition of Selete Pthogens in Rw Beef Met Ysser Shhzi, Nssim Shvisi * Deprtment of Foo Hygiene n Qulity Control, Fulty of Veterinry Meiine, Rzi University, Kermnshh, Irn A R T I C L E I N F O Artile type: Originl rtile Artile History: Reeive: 12 Aug 2018 Aepte: 20 Sep 2018 Pulishe: 22 Sep 2018 Keywors: Beef Met Listeri monoytogenes Oregno Extrt Slmonell typhimurium Stphyloous ureus A B S T R A C T Introution: Listeri monoytogenes, Slmonell spp. n Stphyloous ureus hve een foun in rw met n met prouts. The present stuy ime to evlute the effets of oregno extrt (OE) t the onentrtions of 0.1%, 0.2%, n 0.5% ginst Listeri monoytogenes, Slmonell typhimurium, n Stphyloous ureus in rw eef met uring refrigerte storge (4±1ºC) for 10 ys. Methos: In this stuy, 10 grms of rw eef met were e to 90 milliliters of sterile uffere peptone wter n homogenize in stomher for two minutes. Proper eiml ilutions were prepre in uffere peptone wter for miroil enumertion. S. typhimurium ws ulture on Slmonell Shigell gr n inute t the temperture of 37ºC for 24 hours. In ition, L. monoytogenes ws ulture on PALCAM listeri-seletive gr n inute t the temperture of 30ºC for 48 hours, n S. ureus ws ulture on Bir-Prker gr n inute t the temperture of 37ºC for 48 hours. Results: In ll the experiments, no growth ws oserve in L. monoytogenes, S. typhimurium, n S. ureus in the non-inoulte ontrol smples. Antimiroil effets were more signifint t the higher onentrtions of OE (P<0.05). Moreover, the ition of OEs to rw eef met erese the miroil ounts of the teste pthogens uring storge (P<0.05). Conlusion: Aoring to the results, utiliztion of OE in rw eef met my e n effetive strtegy to ssure sfety ginst L. monoytogenes, S. typhimurium, n S. ureus. However, the ssoite sensory limittions o not llow the use of OE t higher onentrtions, whih re more effetive in pthogen inhiition. Plese ite this pper s: Shhzi Y, Shvisi N. Effets of Oregno Extrt on the Inhiition of Selete Pthogens in Rw Beef Met. J Nutrition Fsting Helth. 2017; 6(1): Doi: /jnfh Introution Proution n hnling of rw met n met prouts epens on nturl ontmintion y environmentl miroorgnisms. Contmintion my our uring slughtering n inreses uring the mnufturing, hnling, pkging, n storge of the prouts (1, 2). Rw met n met prouts re reognize s miroiologilly sfe prouts, n the sfety ssurne epens on the pproprite nti-pthogen effetiveness of multiple ntiteril prmeters se on hurle tehnology (3, 4). However, onsiering the initil ontmintion of rw prouts with high levels of pthogeni teri n inpproprite ontrol of the ntimiroil prmeters, the sfety of these mterils my e ompromise (5, 6). Mny reserhers hve reporte the isoltion of pthogeni miroorgnisms, suh s Listeri monoytogenes, Slmonell spp., n Stphyloous ureus, in rw met n met prouts (3, 7-9). The glol uren of the ssoite foo-orne iseses is signifint, n it is estimte tht only in the Unite Sttes of Ameri (USA), pproximtely 48 million people quire these iseses, 128,000 re hospitlize, n 3,000 ie ue to foo-orne iseses every yer (10). Plnt essentil oils n extrts hve * Corresponing uthor: Nssim Shvisi, Deprtment of Foo Hygiene n Qulity Control, Fulty of Veterinry Meiine, Rzi University, Kermnshh, Irn. Tel: ; Emil: nssim.shvisi@yhoo.om 2017 mums..ir All rights reserve. This is n Open Aess rtile istriute uner the terms of the Cretive Commons Attriution Liense ( whih permits unrestrite use, istriution, n reproution in ny meium, provie the originl work is properly ite.

2 exhiite remrkle ntiteril potentil ginst spoilge n pthogeni teri in fresh met n met prouts (11, 12). Essentil oils n extrts of romti plnts re mong the most importnt tive onstituents of hers n spies (13). Furthermore, their pplition hs inrese in the foo inustry s flvoring ompouns in orer to voi the use of tritionl hemil preservtives (6). The ifferenes in the ntimiroil tivities of vrious herl extrts re usully ssoite with their iverse hemil omposition, whih hnges epening on the seson, geogrphil lotion of the plnts, n the methoology use for plnt extrtion (13). In the pst ees, the onsumer emn for fresh foo prouts hs risen. Negtive pereptions of onsumers towr hemil foo preservtives hve use nturl preservtion tehniques n nturl itives to reeive gret ttention from the foo inustry (8). Non-phytotoxi extrts re onsiere to e sfe foo itives n hve een ertifie s Generlly Reognize s Sfe (GRAS), whih is hs resulte in higher onsumer eptility (11). Oregno is reognize s remrkly effetive herl extrt/essentil oil owing to its ntiteril pility (14). Severl stuies hve use oregno or its onstituents (e.g., rvrol n thymol) s ntiteril gents ginst spoilge n pthogeni teri in fresh met n met prouts (14, 15), vegetles (16), n fresh fish n ephlopos (17, 18). The present stuy ime to evlute the effets of oregno extrt (OE) t the onentrtions of 0.1%, 0.2%, n 0.5% ginst L. monoytogenes, Slmonell typhimurium, n S. ureus in rw eef met uring hille storge (4±1ºC) for 10 ys. Mteril n methos Experimentl Mterils In the present stuy, oregno ws otine from Kermnshh, Irn. The speies of the plnt ws uthentite y Dr. Seyye Ali Msoumi (plnt txonomist) t Rzi University in Kermnshh. The hemil solutions n ulture mei were of nlytil gre n purhse from Sigm (Unite Kingom) n Merk (Germny), respetively. Extrtion Proeure The oregno plnt ws rinse with tp wter, ir-rie for two weeks t mient temperture, n powere using Moulinex foo proessor. Initilly, five grms of the groun plnt ws extrte on mgneti stirrer (IKA, Germny) using pure methnol (10 ml) s the solvent for 24 hours in the rk t mient onitions (25±1ºC). The extrt ws psse through Whtmn gre 4 filter pper, n the resiue ws extrte gin n evporte uner pressure elow the temperture of 40ºC in orer to remove the solvent. Afterwrs, it ws preserve uner hille storge (4±1ºC) efore further use (19). Miroorgnisms n Growth Conitions The exmine strins in this stuy were L. monoytogenes (ATCC 19118), S. typhimurium (ATCC 14028), n S. ureus (ATCC 6538). All the miroorgnisms were preserve t the temperture of -20ºC in Trypti soy roth ontining 30% (v/v) glyerol n suulture twie (7 log CFU/g) efore use in the ssys (20). Preprtion of Rw Beef Met The semimemrnosus musle ws ut from the eef rss within 24 hours fter slughter t lol slughterhouse in Kermnshh (lote in the west of Irn), n the smple ws trnsferre to the foo hygiene lortory uner hille storge (4±1ºC) within 30 minutes. Before the experiments, eh of the met smples ws sterilize for one hour using ultrviolet light in lminr flow hoo, whih ws ivie into four thes of ontrol (without OE) n three thes tht were trete with the OE onentrtions of 0.1% v/w, 0.2% v/w, n 0.5% v/w. Seleting 0.1% s the lowest onentrtion of OE in the urrent reserh ws se on our preliminry stuy. Aoringly, lower OE onentrtions thn 0.1% h no signifint effet on the shelf life extension of the smples. At the next stge, the smples were ippe in the OE solutions (0.1%, 0.2%, n 0.5%) for five minutes t mient onitions through gittion with shker in orer to ensure the omplete istriution of the herl extrt. Following tht, the smples were ir-rie for 15 minutes to llow extrt tthment onto the smples (21). 16 J Nutrition Fsting Helth. 2017; 6(1):

3 The smples were suivie into four thes, inluing th one (without inoulte pthogeni teri), th two (inoulte with S. ureus), th three (inoulte with S. typhimurium), n th four (inoulte with L. monoytogenes). Eh th ompose of six thes (100 g), eh of whih h three portions. For the inoultion of the pthogeni teri, 1000 µl of eh teril suspension (ontining 7 log CFU/ml) ws inorporte into two smples of rw eef met (100 g) seprtely n ompletely mixe y stomher to ensure the uniform istriution of the pthogeni miroorgnisms. Non-inoulte n inoulte smples were pke seprtely in stomher gs, followe y storge t the temperture of 4ºC for 10 ys (8). Miroiologil Anlysis At this stge, 10 grms of rw eef met were inorporte into 90 milliliters of sterile uffere peptone wter n homogenize in stomher (Intersiene BgMixer 400, Nom l Bretèhe, Frne) for two minutes. In ition, suffiient eiml ilutions were prepre in uffere peptone wter for miroil enumertion, inluing S. typhimurium on Slmonell Shigell (SS) gr inute t 37ºC for 24 hours, L. monoytogenes on PALCAM listeri seletive gr inute t 30ºC for 48 hours, n S. ureus on Bir-Prker gr inute t 37ºC for 48 hours (3). All the nlyses were performe on ys zero, two, four, six, eight, n ten in triplite. Sensory Anlysis A pnel inluing 12 juges within the ge rnge of yers ol ws involve in the sensory nlysis. Prior to the nlysis, the pnelists were trine on the sensory properties of rw eef met. Sensory prmeters (oor, olor, n overll eptility) of the smples were evlute using se on 10-point esriptive sle within sore rnge of 1-10 (1=Extremely Dislike, 10=Extremely Like). All the nlyses were performe on ys zero, two, four, six, eight, n ten in triplite (9). Sttistil Anlysis Dt nlysis ws performe in SPSS version 16 using the nlysis of vrine to ssess the effets of vrious OE onentrtions n time of storge on pthogens. Moreover, Dunn s multiple rnge test ws use to ompre the men vlues. In ll the sttistil nlyses, P-vlue of less thn 0.05 ws onsiere signifint. Results&Disussion Figures 1-3 respetively epit the ounts of S. ureus, S. typhimurium, n L. monoytogenes in the rw eef met smples uring 10 ys of 6 S. ureus (log CFU/g) Control OE 0.1% OE 0.2% OE 0.5% Dy Figure 1. Effet of Oregno Extrt (OE) on Survivl of Stphyloous ureus in Rw Beef Met uring Refrigerte Storge J Nutrition Fsting Helth. 2017; 6(1):

4 Control 4 OE 0.1% 3.5 OE 0.2% 3 OE 0.5% Dy Figure 2. Effet of OE on Survivl of Slmonell typhimurium in Rw Beef Met uring Refrigerte Storge S. Typhimurium (log CFU/g) 8 L. monoytogenes (log CFU/g) Control OE 0.1% OE 0.2% OE 0.3% Dy Figure 3. Effet of OE on Survivl of Listeri monoytogenes in Rw Beef Met uring Refrigerte Storge (Eh smple ws nlyze in triplite. On eh smpling y, lowerse letters - inite signifint ifferenes.) (P<0.05). refrigerte storge with n without OE (0.1%, 0.2%, n 0.5% onentrtions). In ll the experiments, no L. monoytogenes, S. typhimurium, n S. ureus growth ws oserve in the non-inoulte ontrol smples (t not shown). Furthermore, higher onentrtions of OE were ssoite with inrese ntiteril tivity (P<0.05). In generl, the ition of OE to rw eef met erese the miroil ounts of the teste pthogens uring refrigerte storge (P<0.05). The tivity of vrious onentrtions of OE ginst S. ureus is shown in Figure 1. The inhiitory effet of OE t the onentrtion of 0.5% ws oserve t the eginning of storge, with the reution of 1 log CFU/g. However, t lower onentrtions, the reution ws only signifintly visulize t the en of storge, with smller logrithmi reution ompre to the ontrol group (1-2 log CFU/g) (P<0.05). In stuy in this regr, Pesvento et l. (22) 18 J Nutrition Fsting Helth. 2017; 6(1):

5 reporte the reution of 1.5 n 2.5 log CFU/g for S. ureus in mine met t the OE onentrtions of 0.5% n 2%, respetively fter 14 ys of storge. On the other hn, only the higher onentrtions of the extrt were effetive for S. ureus. Regring the low onentrtions of OE, our finings were onsistent with the stuy y Grí-Diez et l. (23). In the mentione reserh, no S. ureus inhiition ws enote in houriço ( tritionl pork susge) with 0.05% n 0.005% of OE n sil n trrgon extrts. Moreover, the previous stuies in this regr hve inite tht the pplition of vrious OEs oul ensure the reution of 2-3 log CFU/g in the ount of S. ureus in vrious foo moels, whih is in line with the results of the present stuy (7, 8, 24, 25). Our finings regring S. typhimurium re illustrte in Figure 2-A. Aoringly, S. typhimurium grully erese over time t ll the OE onentrtions. At the onentrtion of 0.5%, n immeite reution ws oserve fter 48 hours, whih ws estimte t 0.8, 1, n 2 log CUF/g on ys four n eight, respetively. After 10 ys, S. typhimurium rehe 2.2 log CFU/g. As for the other onentrtions, the reution of S. typhimurium ws verifie to e more ontinuous over time, n reution of ~1-2 log CFU/g ws lso otine. After 10 ys, reution of 2 n 2.5 log CFU/g ws oserve t the OE onentrtions of 0.1% n 0.2%, respetively (P<0.05). The survivl of S. typhimurium ws lerly ffete y the ition of OE; t ll the investigte onentrtions of the extrt, the survivors erese through the storge perio (P<0.05). Similrly, the finings of Bros et l. (26) emonstrte tht the 0.9% onentrtion of OE resulte in the reution of 1 log CFU/g for S. enteritiis. In ontrst, Grí-Diez et l. (23) reporte no effets ginst S. enteritiis t the 0.005% onentrtion of OE in fermente met susge. For L. monoytogenes (Figure 3), our finings inite the inhiitory tivity of the higher onentrtion of OE (0.5%) t the outset of storge, with logrithmi reution of ~1 log CFU/g, wheres t lower onentrtions, the effet took longer to e visulize on y four of storge. Severl stuies hve expose L. monoytogenes to vrious onentrtions of OE, reporting reutions in miroorgnism ounts. For instne, Dussult et l. (27) pplie the 0.05% onentrtion of OE in hm, reporting reution of 1.5 log CFU/g fter 20 ys. In ition, Tsigri et l. (28) use the onentrtion of 0.8% in met, oserving reution of 2-3 log CFU/g fter 14 ys of refrigerte storge t the tempertures of 2 n 10ºC. In nother stuy, Pesvento et l. (22) use the onentrtions of 0.5% n 2% in mine met, n hieve 1.5 n 2.5 log CFU/g reutions fter 14 ys, whih is in ongruene with the finings of the urrent reserh. Previous stuies hve investigte the ntiteril tivity of OE ginst some fooorne pthogeni teri in-vitro (23, 27, 29), n growing numer of stuies re eing performe on its inhiitory effets in-vivo on vrious foostuffs, inluing rw met n met prouts (23, 30). Aoring to the literture, the min onstituents of OE re phenoli ompouns, thymol, rvrol, p-ymene, linlool, γ-terpinene, β-pinene, terpinen, limonene, n orneol (11, 23, 31). The mehnisms of the omine tivities of ntiteril ompouns in ifferent essentil oils hve not een thoroughly stuie. In reent yers, numer of reserhers hve exmine the mehnism of the ntiteril effets of herl essentil oils, onluing tht the lipophili nture of these ompositions results in importnt morphologil mges, isturing the phospholipi ilyer memrne struture n inresing the permeility of the ell memrne, whih in turn le to the lekge of intrellulr proteins n eletrolytes (11). Sensory Evlution Figure 4 shows the sensory ttriutes of the trete n untrete rw eef met smples, inluing oor, olor, n overll eptility, fter 10 ys of storge. Aoringly, the ontrol smples h signifintly lower sensory ttriutes ompre to the trete smples (P<0.05), whih oul e ue to high miroil growth n hemil hnges (e.g., lipi oxition) (12). Therefore, it oul e onlue tht the inorportion of OE (0.1% n 0.2%) signifintly inrese the sensory properties of the rw eef met smples ompre to the ontrols (P<0.05). The high oor ttriute in the J Nutrition Fsting Helth. 2017; 6(1):

6 Figure 4. Effet of OE on Sensory Properties (oor, olor, n overll eptility) of Rw Beef Met uring Refrigerte Storge (Eh smple ws nlyze in triplite. For eh ttriute, lowerse letters - inite signifint ifferenes.) (P<0.05) trete smples ws not only ue to lower miroil growth ut lso ssoite with the flvor-ontriuting ompouns (e.g., phenols n flvone) (17). Conlusion Aoring to the results, utiliztion of OE in rw eef met oul result in n interesting metho to ssure sfety ginst L. monoytogenes, S. typhimurium, n S. ureus. However, sensory limittions my not llow its use t high onentrtions, whih re more effetive in pthogen inhiition. In the present stuy, it ws onlue tht the high onentrtion of OE (0.5%) erese the ounts of L. monoytogenes, S. typhimurium, n S. ureus in rw eef met. At lower onentrtions (0.1% n 0.2%), lthough the reution ws lower, it ws onsiere signifint for L. monoytogenes, S. typhimurium, n S. ureus. The present stuy my e onsiere strting point for further investigtions fousing on methos to msk the unplesnt sensoril onsequenes use y OE in rw met n met prouts. Moreover, suh investigtions oul ssess the omintion of the lower onentrtions of essentil oils with other tehnologies so s to hieve lne etween the miroil sfety 20 n sensory eptility of rw eef met. Aknowlegments Herey, we exten our grtitue to Rzi University for the provision of filities n instrumenttions in this reserh projet. Conflit of interest None elre. Referenes 1. Lekjing S. A hitosn-se oting with or without love oil extens the shelf life of ooke pork susges in refrigerte storge. Met Si. 2016; 111: Dehn D, Mirzei H, Emm-Djomeh Z, Jfri SM, Dshi S. Therml n ntimiroil properties of hitosn nnoellulose films for extening shelf life of groun met. Crohyr Polym. 2014; 109: Shhzi Y, Krmi N, Shvisi N. Effet of Menth spit essentil oil on hemil, miroil, n sensory properties of mine mel met uring refrigerte storge. J Foo Sfety. 2018; 38(1):e Turgut SS, Soyer A, Işıkçı F. Effet of pomegrnte peel extrt on lipi n protein oxition in eef metlls uring refrigerte storge. Met Si. 2016; 116: Siriptrwn U, Noiph S. Ative film from hitosn inorporting green te extrt for shelf life extension of pork susges. Foo Hyrooll. 2012; 27(1): Khezrin A, Shhzi Y. Applition of J Nutrition Fsting Helth. 2017; 6(1):

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