Article
Evaluation of usage of essential oils instead of spices in meat ball formulation for controlling Salmonella spp. Seda Ozdikmenli1 and Nukhet N Demirel Zorba2
Abstract The purpose of this study was to show the efficacy of essential oils (EOs) in meat balls instead of spices because of their high antimicrobial effect and to evaluate the antimicrobial effect of Origanum onites and Ocimum basilicum EOs against Salmonella Typhimurium in minced beef (20% fat) stored at 4 C for seven days. This is the first study about use of O. basilicum EO in minced beef against bacterial pathogens. Both EOs inhibit microorganisms in in vitro antibacterial tests. Minimum inhibitory concentration (MIC) values of EOs were determined. The lowest MIC values were obtained with O. onites EO 0.6 ml/ml against S. Typhimurium strains. The MIC values of O. basilicum EO 0.25 ml/ml against microorganisms. Both EOs showed a significant decrease in microorganisms inoculated in minced beef at end of storage. The concentration of the both EOs at 20 mg/mg and 10 mg/mg showed stronger antimicrobial activity against bacterial cocktail of S. Typhimurium in beef; however, the higher concentrations caused alterations in the organoleptic properties of meatballs. The results of the present study indicate that O. onites and O. basilicum EOs may be used in combination with each other and different food preservation systems in meat ball formulation.
Keywords Salmonella Typhimurium, Origanum onites, Ocimum basilicum, antimicrobial activity Date received: 9 October 2014; revised: 23 December 2014; accepted: 7 January 2015
INTRODUCTION Meat balls (Kofte) are usually prepared from fresh minced meat and different spices. It is produced in different shapes and consumed widely in all parts of Turkey. Their ingredients change in different regions of Turkey. There are about 300 kind of meat balls such as; Akcaabat, Sultanahmet, Tekirdag, Cig kofte, Satır, and Tire Kofte. This type of food product is very prone to spoil during storage due to its marketing conditions which require meat product to stay fresh. In the meantime, meat products can be exposed to some hazardous conditions to consumer due to its nature which can be contaminated by pathogens like Staphylococcus aureus, Clostridium perfringens, Campylobacter jejuni, Escherichia coli O157:H7, Salmonella, Aeromonas, Listeria monocytogenes, Bacillus cereus, Brochothrix Food Science and Technology International 22(2) 93–101 ! The Author(s) 2015 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/1082013215571118 fst.sagepub.com
spp, and Pseudomonas from different sources such as ingredients and processing conditions (Cetin et al., 2010; Djenane et al., 2011, 2012; Dussault et al., 2014; Gonulalan and Kose, 2003; Klein et al., 2013; Kok et al., 2007; Michalczyk et al., 2012; Oliveira et al., 2013; Solomakos et al., 2008). Many studies were conducted in Turkey about microbiological quality of minced meat and ready to eat meatballs (Baskaya et al., 2004; Cetin et al., 2010; Farag et al., 1989). Cetin et al. (2010) studied 127 samples of ground beef obtained from different regions in Istanbul. They found Salmonella spp. in 4 (3.14%) of the ground beef samples and the mean of aerobic 1 Yenice Vocational School, Canakkale Onsekiz Mart University, Yenice Canakkale, Turkey 2 Department of Food Engineering, Faculty of Engineering, Canakkale Onsekiz Mart University, Canakkale, Turkey
Corresponding author: Nukhet N Demirel Zorba, Department of Food Engineering, Faculty of Engineering, Canakkale Onsekiz Mart University, Canakkale 17020, Turkey. Email:
[email protected]
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Food Science and Technology International 22(2) mesophilic bacteria counts of samples were 1.3 106 CFU/g. Gonulalan and Kose (2003) also found Salmonella spp. in 11% of ground beef samples. They reported that average aerobic mesophilic bacteria counts as 6.0 108 cfu/g. Baskaya et al. (2004) also detected Salmonella spp. in 3 (11%) of minced beef samples. Kok et al. (2007) determined the hygienic quality of cine meatballs and found Salmonella spp. in 18 samples. In addition, Yıldız et al. (2004) analyzed 75 meat ball samples obtained from different regions of Istanbul and found Salmonella spp. in 5.4% of the samples. According to the Turkish Food Codex (Anon., 2009), Salmonella should not be detected in 25 g of meat and meat products. As a results minced meat and meat balls offered to consumers from all part of Turkey have low microbiological quality. Salmonella spp. is leading causes of foodborne bacterial illnesses in humans worldwide. European Union (EU) reports that Salmonella poses a public health risk in minced beef and its preparations (EFSA, 2009). Diseases caused by meat and meat products have the largest share (70%) in the foodborne diseases. Meat industries use synthetic food additives in several meat processes to prevent growth of foodborne pathogens and extend the shelf life of refrigerated storage. However, consumers demand the usage of natural preservatives due to questions about safety of synthetic food additives. In recent years, researchers increasingly have focused on use of natural products like plant essential oils (EOs) as alternative preservatives in foods. EOs are volatile oils known as secondary metabolites of plant parts such as flowers, buds, seeds, leaves, fruits, and roots (Burt, 2004; Lv et al., 2011). Many researchers focused on the antimicrobial and antifungal properties of EOs (Bassole and Juliani, 2012; Baydar et al., 2004; Burt, 2004; Djenane et al., 2012; Farag et al., 1989; Gutierrez et al., 2008; Kalemba and Kunicka, 2003; Karabagias et al., 2011; Klein et al., 2013; Mbandi and Shelef, 2002; Oliveira et al., 2013; Sagdic et al., 2009; Teixeira et al., 2013; Xu et al., 2013). The antimicrobial activity of EOs depends on the chemical composition and how they are produced. Moreover, chemical composition of EOs is influenced by climatic factors, geographical conditions, harvesting, storage, and source of plants (Djenane et al., 2012; Teixeira et al., 2013; Trajano et al., 2010; Xu et al., 2013). Both Origanum onites (Oregano) and Ocimum basilicum (Basil) are some of the oldest members of the Lamiacea (Labiatate) family of spices. It has been known that the EOs of O. basilicum has been used extensively for many years to flavor foods such as confectionaries, baked goods, ketchups, tomato pastes, chili sauces, pickles, vinegar, sausages, and meat products (Bagamboula et al., 2004; Barros et al., 2009; Govaris et al., 2010;
Lachowicz et al., 1998). Many in vitro studies reported a high efficacy of EOs against foodborne pathogens and spoilage bacteria (Burt and Reinders, 2003; Elgayyar et al., 2001; Hammer et al., 1999; Oussalah et al., 2007; Teixeira et al., 2013; Xu et al., 2013). The aim of this study was to demonstrate the efficacy of the EOs instead of spices and, evaluate the antimicrobial effect of O. onites and O. basilicum EOs against Salmonella Typhimurium in minced beef (20% fat) stored at 4 C for seven days.
MATERIALS AND METHODS Preparation of EOs O. onites and O. basilicum EOs were obtained from Agricultural Climatisation and Technological Research Corporation (Ankara, Turkey). According to the manufacturer’s data, plants growing in the West Anatolia region of Turkey were used in the production of the EOs. The oils were isolated by hydrodistillation, and their compositions were determined by Anadolu University, Plant Drug and Scientific Research Centre (AUBIBAM, Eskisehir, Turkey). Identification of individual compartments was done using Wiley/NBS Registry of Mass Spectral Database, NIST MS search, and literature. The EO compounds were identified by matching the peaks of the mass spectra with those of MS libraries. Microbiological and chemical analysis of minced beef Fresh minced beef samples used in this study were provided from a local market (Tesco Kipa Market, Canakkale, Turkey) and were stored at 18 C in a deep freezer (Beko, Istanbul, Turkey). The protein, fat, and moisture contents of the minced beef samples were determined according to Association of Analytical Communities (AOAC) (2000). Enumeration of aerobic mesophilic bacteria, and Enterobacteriacae, was performed by the pour plate technique from appropriate dilutions of homogenized samples on Plate Count Agar (PCA, Merck, Darmstadt, Germany) incubated at 30 C for 48 h and on Violet Red Bile Glucose Agar (VRBG, Merck) incubated at 37 C for 24 h, respectively. Detection of Salmonella spp. was made according to the FDA method (Peter, 2014). Bacterial strain, culture conditions, and preparation of inoculums The EOs were tested against S. Typhimurium (ATCC 14028) and S. Typhimurium (ATCC 51812). To prepare the inoculum, 1 ml of the each culture was transferred separately to 10 ml of sterile tryptic soy broth
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Ozdikmenli and Demirel Zorba (TSB, Merck). Each inoculated broth was incubated at 37 C for 24 h for microbial activation. A standard curve was prepared to establish a correlation between the colony forming unit concentration (CFU/ml) and the optical density (OD) of the dilutions at 625 nm, The cultures were harvested at the mid log phase (37 C for approximately 2 h for Salmonella spp.), then pelleted twice by centrifugation at 20 C and 5000 r/min for 15 min and washed in 0.85% NaCl. The final cell pellet was resuspended in 5 ml of 0.85% NaCl, and the OD was measured at 625 nm to determine the log CFU/ml corresponding to the OD of the culture. Viable cell counts were performed in duplicate by plating serial dilutions in BSA (Merck) for Salmonella spp. incubated at 37 C for 24 h. For the preparation of the bacterial cocktails, 1 ml of culture from each Salmonella strain was transferred to 10 ml of sterile TSB and incubated at 37 C for 24 h for activation, then 2.5 ml of each activated culture was transferred to 50 ml of sterile TSB, and the mixture was incubated at 37 C for 1.5 h (the culture mixture reached to 8 log level within 1.5 h). The cultures were harvested, and the same OD method was used to determine the concentration. Prepared Salmonella cocktails were used in the minced beef inoculations immediately after incubation. Determination of the minimum inhibitory concentration (MIC) The MIC of the EOs against the tested microorganisms was determined using the agar dilution (sensible or non-sensible, þ/) method (National Committee for Clinical Laboratory 402 Standards, 1999) with minor modification; all tests were performed in Mueller Hinton agar (MHA, Merck) supplemented with dimethylsulfoxide (DMSO) (Sigma AldrichÕ -Quimica, Madrid, Spain). (The highest final conc. 3%.) The final concentration ranges of the EOs were from 0.3 ml/ml to 20 ml/ml. This range was chosen based on literatures (Busatta et al., 2008; Careagaa et al., 2003; Djenane et al., 2012; Govaris et al., 2010; Hernandez Ochoa et al., 2011; Ipek et al., 2005; Mytle et al., 2006; Shekarforoush et al., 2007; Singh et al., 2003; Solomakos et al., 2008). S. Typhimurium strains were incubated in TSB until reaching 8 log CFU/ml and inoculated to MHA at a level of 6 log CFU/ml medium. Geometric dilutions of the EOs were prepared in DMSO and added to MHA. Bacterial growths in DMSO alone and in MHA alone were evaluated separately as controls. The inoculated plates were incubated at 37 C for 24 h. The lowest concentration of EOs, at which no visible growth of the bacteria (No growth in the plates) was detected, was defined as the MIC (Farag et al., 1989; Oussalah et al., 2007). All experiments were conducted in duplicate.
Sample preparation and inoculation of S. Typhimurium to minced beef Minced beef was divided into 120 0.1 g portions for each concentration of EOs from either O. onites or O. basilicum and placed into a sterile stomacher bag. EOs were added to the minced beef at the following concentrations: 2.5, 5, 10, and 20 mg/mg and the minced beef were then inoculated with the S. Typhimurium cocktail at 6.88 log CFU per gram of minced beef. The high inoculation level was chosen to determine the direct effect of EOs on counts of Salmonella spp. without a drop below the detection level (Bagamboula et al., 2004; Djenane et al., 2012; Oliveira et al., 2013; Solomakos et al., 2008). EO concentrations were chosen according to MIC values obtained in our study (2.5 ml/ml for O. basilicum) and from MIC values of the EOs of other herbs and spices (Bagamboula et al., 2004; Chouliara et al., 2007; Hayouni et al., 2008; Oliveira et al., 2013; Solomakos et al., 2008). The samples were massaged by hand for 5 min to homogenize the cocktails and EOs. The samples were put in sterile high-density polyethylene bags and stored under aerobic conditions at 4 C without light (MIR-253, Sanyo, Osaka, Japan) for seven days. Samples were taken on days 0, 1, 3, 5, and 7. The control samples were prepared using the same method without the EOs. Three independent replicates of the analyses were performed for each treatment. Bacterial enumeration of surviving cells The samples were analyzed by adding 10 0.1 g of minced beef into a sterile stomacher bag containing 90 ml of 0.1% peptone water and homogenized in a stomacher for 90 s at medium speed (Bagmixer, Interscience, Saint Nom, France), and then serial dilutions were prepared using 0.1% peptone water. S. Typhimurium enumeration was performed by spread plate method on bismuth sulfite agar (BSA, Merck). Bacterial colonies were counted after incubation at 37 C for 48 h. Sensory analysis Sensory derived effects of adding each of the studied EOs to meat ball were evaluated by ordering (Ranking classification) of samples according to the appearance, texture, flavor, and bitterness/throat catching (Meilgaard et al., 1999). Both taste and odor of untreated and treated samples were checked. The concentrations of the EOs of O. onites and O. basilicum in the treated samples were 2.5, 5, 10, and 20 mg/mg, and the control sample was prepared with no EO. Samples were prepared as follows: fresh minced beef (300 g) 95
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Food Science and Technology International 22(2) meat was mixed by hand with wetted bread (20%), cumin (0.1%), dry mint (0.1%), salt (1%) (that components have been used to make traditional meatball) and an EO, and then the mixture was formed into a meatball and fried in sunflower oil at 190 C for 2 min and served. White bread and water were provided after each sample to clean the palate. Panelists were selected among the students and staff of the Department of Food Engineering. The samples were also evaluated by an acceptance test using a seven-point hedonic scale from 1—dislike extremely to 7—like extremely. Each of 25 panelist evaluated samples by ranking and rating the different qualities.
STATISTICAL ANALYSIS In total, two EOs (O. onites and O. basilicum) at five concentrations and four storage times were tested for antibacterial activity against S. Typhimurium in minced beef. For each condition, three independent replicates of the experiment were performed, and duplicate samples were analyzed at each sampling time (n: 3 2). The significant difference between the samples was determined by ANOVA. Duncan and Turkey’s multiple range tests were also applied for comparison of means. Data were analyzed using Minitab 15.0 (Minitab 15.0 Minitab Inc., PA). In sensory analysis, the variability of acceptance or liking of samples was analyzed by non parametric Kruskal–Wallis and Dunn test (NCSS, Kaysville, UT, 2007).
RESULTS AND DISCUSSION Composition of the EOs The gas chromatography–mass spectrometry (GC-MS) analysis identified the major constituents (>1%) of the EOs. The compositions of EOs were given in Table 1. The main components of EOs obtained were carvacrol (67.0%) and linalool (40.6%) for O. onites and O. basilicum, respectively (Ozdikmenli and Zorba, 2013). Ipek et al. (2005) and Baydar et al. (2004) found carvacrol as the main component of O. onites EO (74% and 86.9% respectively) while the main component of O. basilicum EO was found as linalool by Lachowicz et al. (1998) and Opalehenova et al. (2002) 44%, 54.95%, respectively. But Ozcan and Chalchat (2002) found methyl eugenol (78.02%) Bagamboula et al. (2004) found estragol (20.50%) as the main component of O. basilicum EO. Amount of EO’s chemical substance was affected by many different factors such as climate, part of plants, harvesting period, and isolation technique (Burt, 2004; Djenane et al., 2011; Lv et al., 2011; Oussalah et al., 2007; Ozcan and Erkman, 2001).
Many researchers reported that major and minor chemical components of EOs performed together against the bacterial cell (Bassole and Juliani, 2012; Burt, 2004; Lv et al., 2011). These components are phenols, terpenes, aldehydes, and ketones, and they disrupt the cell membrane, affect the permeability, and cause leakage of various substances such as ions, ATP, nucleic acids, and amino acids which result in death of cell (Burt, 2004; Lachowicz et al., 1998). Microbial quality and chemical composition of minced beef The average aerobic mesophilic bacteria and Enterobacteriacea counts were determined as 1.90 104 and 1.20 102 CFU/g, Salmonella spp. was not detected in any samples. It was found that protein, fat, and moisture content of minced beef were 18.09 0.75%, 18.89 0.51%, and 62.34 0.96%, respectively. Meat and meat products can have a high fat content and reduce the effect of EOs against bacteria. Furthermore, meat and meat products are rich source of protein, minerals, and vitamins. This structure supports the growth of microorganisms and reduces the action of EOs in meat (Bajpai et al., 2011; Burt, 2004; Hernandez Ochoa et al., 2011). Minimal inhibitory concentration The EOs of O. onites and O. basilicum had significant inhibitory effects on S. Typhimurium in MHA. The O. onites EO showed a higher effect compared to the EO of O. basilicum in the inhibition of microorganisms. The MIC value of O. basillicum was found as 2.50 ml/ml for both S. Typhimurium strains. The MIC values of O. onites for both strains of S. Typhimurium were 0.60 ml/ml. Many researchers reported different MIC values of different Origanum and Ocimum species for certain microorganisms. (Barros et al., 2009; Erturk et al., 2006; Hussain et al., 2008; Souza et al., 2006). Erturk et al. (2006) determined the MIC value of O. onites as 2.35 mg/ml against S. Typhimurium CCM 5445 which is higher than our findings. Thus, antimicrobial effect depends on the amount of the main constituents of the EO. Ozcan and Erkmen (2001) also determined the antimicrobial activity of the EOs of nine plant species at three concentrations, EO of O. basilicum L. was found to be ineffective against S. Typhimurium KUEN 1357 at all concentrations tested (1–15% vol/vol). Rattanachaıkunsopon and Phumkhachorn (2010) reported a MIC value of O. basilicum EO for the inhibition of Salmonella Enteritidis (ATCC 10708) as 40 mg/ ml and linalool and methyl chavicol are the main
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Ozdikmenli and Demirel Zorba Table 1. Composition of Origanum onites and Ocimum basilicium essential oilsa (Ozdikmenli and Zorba, 2013) Ocimum basillicum
Origanum onites
constituents
%
constituents
%
1. 2. 3. 4. 5. 6. 7. 8. 9.
1.0 8.8 40.6 2.3 28.5 1.6 1.1 1.9 2.0
1. Myrcene 2. alfa-Terpinene 3. gama-Terpinene 4. p-Cymene 5. Linalool 6. beta-Caryophyllene 7. Borneol 8. beta-Bisabolene 9. Thymol 10. Carvacrol Total
1.3 1.1 5.6 4.9 6.1 2.0 2.0 1.0 1.7 67.0 92.7
b-Pinene 1,8-Cineole Linalool Trans-b-Bergamotene Methylchavicol Germacrene D g-Muurolene Eugenol T-Cadinol
Total
87.8
a
More than 1%.
constituents of their basil EO like as used basil EO in the present study. Survival of S. Typhimurium strains in EO treated minced beef The results showed that O. onites EO is more effective than O. basilicum EO against S. Typhimurium in minced meat. The compositions of EOs obtained from Oregano and Basil and their antimicrobial effects had been determined in various studies but the results varied from country to country (Bagamboula et al., 2004). Previous researches showed that the addition of EOs to minced beef exerted antimicrobial effect with MIC values higher than those reported in the present study (Burt, 2004; Gutierrez et al., 2008; Juliano et al., 2000; Singh et al., 2003). The effectiveness of EOs is known to be reduced in a food matrix system as a result of the interaction with different components of food (Burt, 2004; Gutierrez et al., 2008; Juliano et al., 2000; Michalczyk et al., 2012; Oliveira et al., 2013; Singh et al., 2003). In our study, the MIC value of O. basilicum EO was chosen as the lowest concentration for two EOs that were added to minced beef and the highest concentration was chosen according to used amounts given in previous studies and according to the differences in the main components of used EOs in this study and previous studies (Juliano et al., 2000; Skandamis and Nychas, 2001). This study demonstrated that the concentrations of 20 mg/mg O. onites and O. basilicum EOs were the most effective concentration against S. Typhimurium on the minced beef meat during seven days. The population of S. Typhimurium decreased in all EOs treated samples.
The O. onites EO and O. basilicum EO exhibit antimicrobial activity against S. Typhimurium at the 10 mg/mg and 20 mg/mg concentrations in minced beef. There was a significant difference between control and O. onites (10 mg/mg and 20 mg/mg), O. basilicum (10 mg/ mg and 20 mg/mg) treated samples (p < 0.05). When the S. Typhimurium increased in control samples O. onites (10 mg/mg and 20 mg/mg) decreased S. Typhimurium counts by 1.18 and 2.87 log CFU/g, respectively. Both 5 mg/mg and 2.5 mg/mg concentrations of EOs also decreased S. Typhimurium counts in minced beef meat (Figure 1). However, present populations of pathogens in samples treated with these concentrations are not significantly different than the samples of control. The interaction effect of storage days and used concentrations of O. onites EOs on populations of S. Typhimurium in samples are statistically significant (p < 0.05). Samples treated with O. basilicum EOs at 20 mg/mg and 10 mg/mg concentrations showed populations of S. Typhimurium significantly lower than the control samples (2.20 and 0.67 log CFU/g) at the end of storage days. The interaction effect of storage days and used concentrations of O. basilicum EOs on the populations of S. Typhimurium in samples are also statistically significant (p < 0.05). Figure 1 shows the survival curves of S. Typhimurium on the minced beef meat during seven days. Govaris et al. (2010) found that oregano EO at 0.9% was more effective against Salmonella Enteritidis than oregano EO at 0.6% on minced sheep meat at 4 C and 10 C for 12 days. They inoculated the meat samples at a level of 4.1 log CFU/g and achieved a total inactivation at 4th storage day when they used oregano EO at 0.9%. Their inactivation rate was
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Origanum onites A
8
Ocimum basilicum B
7 0 µg / mg
6
2.5 µg / mg 5 µg / mg
5
10 µg / mg
4
20 µg / mg
3 0
1
2 3 4 5 Storage time (days)
6
7
S.Typhimurium log CFU/g of meat
S.Typhimurium log CFU/g of meat
Food Science and Technology International 22(2)
8 7
0 µg / mg 2.5 µg / mg
6
5 µg / mg
5
10 µg / mg
4
20 µg / mg
3 0
1
2 3 4 5 Storage time (days)
6
7
Figure 1. Effect of different concentrations (20, 10, 5, 2,5, 0 (control) mg/mg) of Origanum onites (a) and Ocimum basilicum (b) essential oils against Salmonella Typhimurium cocktail in minced meat during storage (4 C 7 days), error bar shows standards errors.
Table 2. Effect of Origanum onites essential oil on sensory characteristics of meat balls Origanum onites Scoresa Concentration(mg/mg)
Appearance
Texture
Odor
Rancidity
Control (0) 2.5 5 10 20 p Values
4.48 1.25A 4.20 1.16A 4.44 1.05A 3.92 1.30A 3.36 1.33A 0.17
4.52 1.25A 3.76 1.23AC 3.84 1.30AB 3.44 1.23BC 2.84 1.12C 0.02
4.60 1.11A 3.36 1.14B 2.68 1.12BC 2.04 0.93CD 2.04 1.04CD 0.01
3.52 1.77A 2.88 1.36A 3.00 1.53A 2.36 1.33A 2.20 1.40A 0.22
Note: Score means within each column with no letters in common are significantly different (p 0.05). a Mean SE.
higher than our rates. This result could be ascribed to usage of lower inoculation level, different kind of oregano, different amounts of main components and difference in resistance of Salmonella species. Tsigarida et al. (2000) reported a reduction in initial micro flora on beef fillets by 2–3 log CFU/g with the addition of 0.8% of oregano EO. Skandamis and Nychas (2001) reported an immediate suppression in the total viable count in minced beef by 1 log CFU/g when oregano oil was added at a concentration of 1% which agrees with the findings of this study. The evaluations given by each of the panelist in the sensory test are summarized in Tables 2 and 3. Each treatment of O. basilicum EO resulted in no significant difference in appearance, while each treatment of O. onites EO resulted in no significant difference in both appearance and bitterness (taste). The O. basilicum EO was found to have significantly different effect on odor, texture, and bitterness compared to the control sample. However, O. onites EO only caused a significant difference in the texture and odor compared to the control
sample. The panelists indicated that the supplementation levels of 5 mg/mg and 2.5 mg/mg were acceptable in O. onites EO while it is unacceptable for all levels of O. basilicum EO. The practical application of several EOs in foods is limited because EOs of spices have a strong flavor. Chouliara et al. (2007) and Tsigarida et al. (2000) found that the flavor of beef fillets treated with 0.8% vol/wt oregano oil was acceptable after storage at 5 C and cooking. It was reported that the flavor, odor, and color of minced beef containing 1% vol/wt oregano oil improved during storage under modified atmosphere packaging and vacuum at 5 C and were almost undetectable after cooking (Skandamis and Nychas, 2001). Although there were some researches which report acceptability of 10 mg/mg oregano EO in minced meat samples, our panelist found 20 mg/mg and 10 mg/mg were more bitter/throat catching than other samples. These differences between their results and our study may depend on EO composition.
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Ozdikmenli and Demirel Zorba Table 3. Effect of Ocimum basilicum essential oil on sensory characteristics of meat balls Origanum onites Scoresa Concentration(mg/mg)
Appearance
Texture
Odor
Rancidity
Control (0) 2.5 5 10 20 p Values
4.48 1.25A 4.20 1.16A 4.44 1.05A 3.92 1.30A 3.36 1.33A 0.17
4.52 1.25A 3.76 1.23AC 3.84 1.30AB 3.44 1.23BC 2.84 1.12C 0.02
4.60 1.11A 3.36 1.14B 2.68 1.12BC 2.04 0.93CD 2.04 1.04CD 0.01
3.52 1.77A 2.88 1.36A 3.00 1.53A 2.36 1.33A 2.20 1.40A 0.22
Note: Score means within each column with no letters in common are significantly different (p 0.05). a Mean SE.
CONCLUSION The results of this study show that O. onites EO was more effective than O. basilicum EO against S. Typhimurium in minced beef meat during storage at 4 C. Meat balls prepared with O. basilicium EOs and more than 5 mg/mg concentration of O. onites EOs were not organoleptically acceptable at the day of preparation when they were fried, however Skandamis and Nychas (2001) reported that the effect of oregano oil on organoleptic properties decreased during storage so additional studies should be done for determination of changes in organoleptic aspects by changing storage period. Bagamboula et al. (2004) suggested that strong aroma associated with EOs should be solved by the use of methods which consist in dearomatization, such as supercritical CO2 extraction. However, it is important to keep antimicrobial properties of EOs during dearomatization (Chaibi et al., 1997). In addition, synergistic combination of O. onites and O. basilicum EOs may be used to decrease their concentrations and to minimize their adverse sensorial effects in meat balls. In general, EOs and their combinations could be more effective by hurdle-effect (Klein et al., 2013). DECLARATION OF CONFLICTING INTERESTS The authors declare that there is no conflict of interest.
FUNDING The authors are grateful to the Canakkale Onsekiz Mart University Scientific Research Foundation for the financial support for this work (BAP 2009/124).
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