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Effect of antimicrobial packaging on physicochemical and microbial quality of chicken drumsticks
Food Control 54 (2015) 294e299
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Food Control journal homepage: www.elsevier.com/locate/foodcont
Effect of antimicrobial packaging on physicochemical and microbial quality of chicken drumsticks dem Soysal a, *, Hüseyin Bozkurt a, Engin Dirican a, Mehmet Güçlü b, Çig Ebru Deniz Bozhüyük b, Ali Erdal Uslu b, Sevim Kaya a a b
Department of Food Engineering, Faculty of Engineering, University of Gaziantep, 27310 Gaziantep, Turkey NAKSAN Plastic Company, 1.Organized Industrial Zone, 83118 Street, Bas¸pınar, 27120, Gaziantep, Turkey
a r t i c l e i n f o
a b s t r a c t
Article history: Received 10 November 2014 Received in revised form 3 February 2015 Accepted 10 February 2015 Available online 18 February 2015
The effects of antimicrobial agents (nisin, chitosan, potassium sorbate (PS) or silver substituted zeolite (AgZeo)) incorporated into low density polyethylene (LDPE) on the physicochemical and microbial quality of chicken drumsticks stored at 5 C for 6 days were investigated. Active multilayer bags (LDPE/polyamide/LDPE-containing 2% antimicrobial agent) were used to pack chicken drumsticks under vacuum. Their efficiency in inhibiting total aerobic mesophilic bacteria (APC), total coliforms and total molds and yeasts were evaluated by comparison with control bag (LDPE-polyamide-LDPE). APC counts of samples packed in bags containing chitosan, nisin, AgZeo and PS were 1.03, 0.98, 0.51, and 0.17 log respectively were lower than those of samples packed in control bags. Samples packaged in active bags had lower microbial counts and thiobarbituric acid reactive substance (TBARS) values than those of samples packed in control bags. The pH, color and hardness of the samples were not affected significantly by use of different packaging films (p > 0.05). Therefore, the bag including chitosan or nisin can be suggested as a suitable packaging material for chicken drumstick to increase safety and quality. © 2015 Elsevier Ltd. All rights reserved.
Keywords: Active packaging Antimicrobial agent Chicken drumstick
1. Intoduction Active package changes the condition of the packed food to extend shelf-life or to improve safety or sensory properties, while maintaining the quality of packaged food (Ahvenainen, 2003; Hutton, 2003). Antimicrobial packaging appears to be one of the most promising applications of active food packaging technology (Joerger, 2007; Quintavalla & Vicini, 2002). The purpose of incorporating antimicrobials into film is to control spoilage as well as contamination with pathogens. Antimicrobial compounds incorporated into the packaging materials may be configured to be slowly released during food distribution and storage (Ishitani, 1995). As a result, microbial quality of packaged food products can be drastically improved and shelf life significantly extended. Commonly studied antimicrobial agents for active packaging are: nisin (Economou, Pournis, Ntzimani, & Savvaidis, 2009; Ercolini et al., 2010; La Storia et al., 2012; Pranoto, Rakshit, & Salokhe, 2005), chitosan (Chen, Weng, & Chen, 1999; Chounou
* Corresponding author. Tel.: þ90 342 317 23 07; fax: þ90 342 3601105. E-mail address: [email protected] (Ç. Soysal). http://dx.doi.org/10.1016/j.foodcont.2015.02.009 0956-7135/© 2015 Elsevier Ltd. All rights reserved.
et al., 2013; Quattara, Simard, Piette, Begin, & Holley, 2000), potassium sorbate (Han & Floros, 1997; Pranoto, et al., 2005; Weng & Hotchkiss, 1993), silver substituted zeolite (Brody, Stupinsky, & Kline, 2001; Ishitani, 1995), triclosan (Cutter, 1999; Vermeiren, Devlieghere, & Debvere, 2002), and essential oils (Hong, Park, & Kim, 2000; Oussalah, Caillet, Salmieri, Saucier, & Lacroix, 2004). Although these antimicrobial materials are active against several individual microorganisms under laboratory conditions, it has proven difficult to produce films that are sufficiently active in food systems (Balasubramanian, Rosenberg, Yam, & Chikindas, 2009). Multilayer packaging systems have been used to increase shelf life and safety of food products. Polymers can provide both improved barrier properties to both gas and water transfer and improved mechanical strength. Different polymers have different properties. For example, polyethylene, which is the most widely used polymer in food packaging, is a good barrier for water but a poor barrier for gases. Polyamide, however, provides a poor barrier for water but a good barrier for gases. So multilayer bags combining both possess higher barriers for both water and gases as well as providing increased mechanical strength. Incorporation of antimicrobial compounds into multilayer packaging polymers will be an alternative for limiting microbial growth and increasing shelf life of
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food products such as poultry meat. Poultry is widely consumed an important source of protein in human diet. Safety and quality of poultry meat are important issues for the food industry and also for consumers. Developing a method to increase shelf-life and overall safety/quality represents a major challenge for the poultry processing industry (Economou et al., 2009). Microbial contamination, which reduces the shelf-life of poultry meat and increases the risk of food borne illness, occurs mainly on the surface of poultry meat during slaughtering, processing, transportation and storage (Massoni, Morando, Vignolo, & Eisenberg, 2012). The objective of the current study was to determine the effect of different antimicrobial agents (chitosan, nisin, PS, AgZeo) on the microbial, physical and chemical characteristics of chicken drumsticks during refrigerated storage, compared with control multilayer bag without antimicrobial agent and having the same thicknesses.
2.3.1. Microbiological analysis Drumsticks were deboned under the aseptic conditions. About 25 g of drumstick sample was weighed aseptically, transferred to a flask containing 225 mL of sterile water and homogenized in a Waring Blender for 15 min at room temperature. For microbial enumerations, 0.2 mL samples of serial dilutions were spread on the surface of petri plates. APC was determined using Plate Count Agar after incubation at 37 ± 1 C for 2 days. Total mold and yeast counts were determined using Potato Dextrose Agar after incubation at 25 ± 1 C for 5 days. For total coliform counts, 0.2 mL sample was spread onto Violet Red Bile Agar and incubated at 37 ± 1 C for 2 days. Large colonies with purple haloes were counted. All plates were examined visually for typical colony types and morphological characteristics associated with each growth medium. Microbiological data are expressed as a logarithm of colony forming units per gram (log cfu/g).
2. Materials and methods
2.3.2. Chemical analysis The pH value was recorded using a pH meter. About 10 g of deboned chicken drumstick sample was homogenized in a Waring Blender with 90 mL of distilled water and the homogenate was used for pH determination. TBARS values of the samples were determined by a spectrophotometric method (Bozkurt, 2006). A 2-g sample was taken and TBARS were extracted twice with 10 mL of 0.4 M perchloric acid. The extracts were diluted to 25 mL with 0.4 M perchloric acid. The extracted solution was centrifuged for 5 min at 1790G. After centrifugation, 1 mL of supernatant was poured into a glass-stoppered test tube, 5 mL of TBA reagent was added and the mixture was heated in a boiling water-bath for 35 min. After cooling the tube in tap water, the absorbance of the sample was read against the appropriate blank at 538 nm. A calibration curve was prepared using malondialdehyde (MDA). TBARS values were expressed as mg of malondialdehyde/kg chicken sample.
2.1. Materials The polymers used in the present study were provided by NAKSAN Plastic Company (Gaziantep, Turkey). Nisin and chitosan were obtained from Handary SA, potassium sorbate and silver substituted zeolites were purchased from Aldrich Chemical Company, Inc., USA. Chicken drumsticks were obtained from a local slaughterhouse. 2.2. Film preparation The active and control bags were produced in NAKSAN Plastic Company. The active LDPE layer was obtained by mixing antimicrobial agents (2% of each chitosan, nisin preparation, PS, or AgZeo) with polyethylene pellets. Firstly, master batches were prepared by addition of 2% antimicrobial agent, 2% orevac as tie layer, 2% ethylene vinil acetate copolymer (EVA), and 94% polyethylene pellets, using a twin screw extruder. These master batches were used in production of active layers of the multilayer films. The active-multilayer (LDPE-polyamide-active LDPE) and control (LDPE -polyamide- LDPE) films having the same thicknesses (70 mm) were produced using a blown film extrusion process. Then films were sealed to prepare bags with 15 cm width and 25 cm length in sterile conditions. Oxygen transmission rates of the films were measured by the following procedure of ASTM D3985-05 (ASTM 2010) using an Ox-Tran Model 2/21 (Mocon, Inc., Minneapolis, Minn., U.S.A). The measurement conditions for oxygen permeability were 23 C, 100/0% relative humidity and 1 atm pressure. Permetran Model C 4/41 (Mocon, Inc., Minneapolis, Minn., U.S.A) was used to measure water vapor transmission rate of the films according to ASTM F1249-13 (ASTM 2013) at 23 C. 2.3. Packaging and storage of samples All chicken drumsticks were collected from the same batch of the slaughtered chickens, with all of them coming from the same farm. Chicken drumsticks were transported under chilled conditions to the laboratory and immediately packaged. The drumsticks were singly vacuum-packaged in active bags and control bags using a manual vacuum sealing machine, and stored at 5 C. For each packaging system, four samples were taken after 0, 1, 2, 3, 4, and 6 days of storage. The pH, color, TBARS values and microbiological analysis (total coliform, APC and mold and yeast counts) of the drumstick samples were followed after removal of the bags.
2.3.3. Physical analysis The changes in skin color (Hunter L*, a* and b*) of the drumsticks packed with active and control bags were determined from the same areas of the samples after removal of the bags, in all measurements. Chicken drumstick skin color was measured using a Hunter lab ColorFlex (A60-1010-615 Model colorimeter, Hunter lab, Reston, VA). The instrument was standardized each time with a white and black ceramic plates (Lo* ¼ 93.01, ao* ¼ 1.11 and bo* ¼ 1.27), and L*, a* and b* values of samples were recorded as average of four readings taken at same locations on each chicken drumstick. Hardness values of the chicken drumstick samples were determined using TA-XT2 Texture analyzer (Texture Technologies Corp., Scarsdale, NY/Stable Microsystems, Godalming, UK). Hardness of the samples was measured using a needle probe (1.8 mm in diameter) and recorded after penetration of the needle to a depth of 5 mm into sample at a speed of 1 mm/s. Data collection and calculation were done using Texture Expert Exceed Version 2 V3 (Stable Micro Systems, 1998). The procedure was repeated for two different samples and three times on each sample. The hardness is given as the mean of six measurements and expressed as Newton (N). All hardness values were determined from the same region of the samples. 2.4. Statistical analysis Analysis of Variance (ANOVA) was performed for each response (mold and yeast count, APC, total coliform count, pH, TBARS, L*, a*, b*, and hardness) to determine significant differences using the SPSS version 16.0 (SPSS Inc., Chicago, IL, USA). Duncan's multiple range tests were also carried out to distinguish examined groups.
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3.2.1. Microbiological quality APC, total coliform and mold and yeast counts are used by the poultry industry as general indicators of processing hygiene, storage quality and potential shelf-life (Capita, Alonso-Calleja, Garcia Fernandez, & Moreno, 2001; Alvarez-Astorga, Alonso-Calleja, ndez, 2002). Changes in APC (log Capita, Moreno, & Garcia-Ferna cfu/g) in chicken drumstick packaged with the active and control bags during 6 days of storage at 5 C are shown in Fig. 1. APC of all samples increased (P < 0.05) during storage and at the end of the storage period, APC of samples ranged from 4.70 to 5.73 log cfu/g. Similar results were reported by Kim and Marshall (1999) who found that APC in chicken leg increased to about 6 log cfu/g after 6 days of storage at 4 C. In our samples, the increase in APC in chitosan-incorporated packaging was about 0.5 log cfu/g whereas it was 1.53 log cfu/g in samples stored in control bags. The increases in APC were very low when compared with the data reported by Del Rio, Panizo-Mor an, Prieto, Alonso-Calleja, and Capita (2007) who reported a 6-log cfu/g increase in APC of chicken leg samples (from about 3.20 to 9.20 log cfu/g) after 5 days of storage at 3 C. Cox, Bailey, and Berrang (1998) reported that spoilage of poultry generally occurs when total aerobic mesophilic counts exceed 7 log cfu/g. In the present study, all drumsticks samples had lower than 6 log cfu/g after 6 days of storage.
Chicken drumsticks packaged with active bags had lower (P < 0.05) APC than the sample packed with control bags. At the 6th day of storage, APCs of samples packaged with nisin or chitosan bags were not statistically different (P > 0.05), but were lower (P < 0.05) APC than those of other samples. Similar results were also found by Siragusa, Cutter, and Willett (1999) and Park, Marsh, and Dawson (2010). Siragusa et al. (1999) used nisin incorporated low density polyethylene (LDPE) film for controlling food spoilage and enhancing product safety since nisin, a bacteriocin, inhibits Grampositive food borne pathogens and spoilage microorganisms. Park et al. (2010) observed that chitosan could be released from LDPE films to inhibit bacterial growth and that higher chitosan concentrations (1.4%) inhibited both gram positive and gram negative bacteria. The antibacterial property of chitosan is well known and is likely due to the interaction of the positively charged chitosan with negatively charged residues on the cell surface of many fungi and bacteria, causing extensive cell surface alterations and altering cell permeability. Changes in total coliform count in chicken drumstick packaged with control and active bags are shown in Fig. 2. Total coliform count increased (P < 0.05) during the 6 days of storage at 5 C from 3.19 log cfu/g to the counts ranging from 3.82 to 4.57 log cfu/g. Kim and Marshall (1999) observed that total coliform counts increased in chicken legs during storage and Del Rio et al. (2007) found that total coliform count increased from about 2.10 log cfu/g to about 7 log cfu/g during storage for 5 days at 3 C. In our study, the increase in total coliform count was lower (as about 1.4 log cfu/g) than that reported by Del Rio et al. (2007). Alvarez-Astorga et al. (2002) found that the contamination level of legs with coliforms was around 3.56 log cfu/g and reported that levels of coliforms in chicken skin varied between 2.7 and 4.9 log cfu/g. No reduction in Escherichia coli cells was observed in LDPE film containing chitosan polymer or oligomer (Hong et al., 2000) in a cast corn zein film containing lysozyme (Padgett, Han, & Dawson, 1998), and in an LDPE film coated with nisin (An, Kim, Lee, Paik, & Lee, 2000). However, in our study, use of active bags containing chitosan and nisin decreased (P < 0.05) total coliform count of chicken drumstick when compared with the control sample. The lowest (P < 0.05) total coliform count was observed in the chicken drumstick packed with chitosancontaining film whereas the highest counts (P < 0.05) was obtained with chicken drumstick packed in control bags. One-way ANOVA showed that mold and yeast counts were affected by both storage time and incorporation of antimicrobial
Fig. 1. Effects of antimicrobial packages on APC (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
Fig. 2. Effect of antimicrobial packages on total coliform counts (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
3. Results and discussions 3.1. Barrier properties of films Water vapor and oxygen transmission rates are important for controlling the barrier properties of films. Water and oxygen can accelerate the growth of microorganism and thus reduce the shelf life of foods. Water vapor transmission rates (g/m2$day) and oxygen transmission rates (cc/m2$day) of control bags were determined as 10.10 and 20.9, respectively. Addition of antimicrobial agents had no significant effect on the water vapor transmission rates of active bags which were found to be 9.55, 7.95, 8.15 and 7.85 g/m2$day for chitosan, nisin, PS and AgZeo added bags, respectively. However, oxygen transmission rates of active bags had significantly affected (P < 0.05) by the addition of chitosan (9.3), nisin (7.5), PS (7.0) and AgZeo (7.8 cc/m2$day). 3.2. Effect of antimicrobial packaging on quality of chicken drumsticks
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agents into the plastic bags. Mold and yeast count increased (P < 0.05) from 5.49 to levels ranging from 5.84 to 6.86 log cfu/g after 6 days of storage. As shown in Fig. 3, the drumsticks packed with active bags had lower (P < 0.05) mold and yeast counts than those packed in control bag. The rate of increase in mold and yeast counts during the storage periods was the highest in drumstick packed in control bags whereas it was the lowest in the samples packed in chitosan-containing bag. As a result, samples packaged with antimicrobial agents, especially with chitosan, nisin and PS, had low mold and yeast count throughout the 6 days of storage period. The efficiency of using chitosan-incorporated polyethylene extruded films in inhibiting radial growth of Aspergillus niger has been reported by Martínez-Camacho et al. (2013). There are conflicting results in the literature regarding the activity of PS, but one report suggested that the incorporation of PS into polyethylene films (0.4-mm thick) lowered the growth rate of mold and yeast (Han & Floros, 1997). Some authors stated that using low density polyethylene films (0.05-mm thick) containing 1.0% w/w sorbic acid were unable to suppress mold growth (Weng & Hotchkiss, 1993). Moreover, ethylene vinyl alcohol/linear low-density polyethylene (EVA/LLDPE) film (70 mm thick) impregnated with 5.0% w/w PS was unable to inhibit the growth of microorganisms on cheese (Devlieghere, Vermeiren, Bockstal, & Debevere, 2000). Results of the present study indicated that active bags have an inhibitory effect on microbial growth in chicken drumsticks when compared with the control. Table 1 shows the relative decrease in microorganisms (in log cfu/g) of samples packaged in active bags with respect to samples packaged in control bags at the end of the storage period. Chitosan was the most effective active bag with respect to APC, total coliform and mold and yeast counts. Nisin was also effective on inhibiting the APCs, total coliform and mold and yeasts. PS was found as effective on mold and yeast counts of chicken drumsticks. Although the microbial counts of samples packed with AgZeo were lower than the control samples, it was the least effective one around the studied antimicrobials.
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Table 1 Reduction in microbial counts (log cfu/g) of chicken drumstick in antimicrobial bags with respect to control bags at the end of storage period. Chitosan ± SE Nisin ± SE APC 1.03 ± 0.009 Mold and 1.02 ± 0.009 yeast Total 0.75 ± 0.104 coliform
PS ± SE
AgZeo ± SE
P-Value
0.98 ± 0.019 0.17 ± 0.060 0.51 ± 0.024 <0.01 0.87 ± 0.020 0.94 ± 0.004 0.25 ± 0.030 <0.01 0.68 ± 0.047 0.25 ± 0.163 0.28 ± 0.118 <0.05
Note: At the end of the storage period in counts of APC is 5.73 log cfu/g; mold and yeast is 6.86 log cfu/g; total coliform is 4.57 log cfu/g in control.
3.2.2. Chemical quality Lipid oxidation is a major contributor to quality deterioration process in muscle foods, resulting in a variety of breakdown products which produce undesirable off-odors and flavors. The initial TBARS value of the drumstick was 1.32 mg/kg sample. The suggested limit for TBARS value in meat products is 2 mg/kg above which rancid off-flavors become sensorially detectible (Byun et al.,
2003). After 2 days of storage, the TBARS values of the samples except chitosan were exceeded TBARS value of 2 mg/kg (Fig. 4). Although an increasing trend of TBARS values of the all samples was detected, there was no significant difference between those of the control samples and the samples packed with active bags, up to 2 days (p < 0.05). After 2 days of storage, TBARS values of all samples packaged with active bags were significantly lower (p < 0.05) than those of the samples packaged with control bags. Lower rate of lipid oxidation in samples packed with active bags compared to samples packed with control bags can also be related with the lower oxygen transmission rates of active bags compared to control bags. The susceptibility of drumstick meat towards oxidation has been explained by the high content of polyunsaturated fatty acids in this muscle (Chouliara, Karatapanis, Savvaidis, & Kontominas, 2007). Lowest lipid oxidation was observed for samples packed with chitosan incorporated bags, which might be helpful to limit lipid oxidation of the drumsticks during the storage period. The above results are in agreement with those of Georgantelis, Ambrosiadis, Katikou, Blekas, and Georgakis (2007) who reported a reduction in TBARS values for pork sausages by 73% for samples containing 1% chitosan (0.25 mg MDA/kg) as compared to control samples (0.96 mg MDA/kg after 10 days of storage at 4 C). Darmadji and Izumimoto (1994) reported that TBARS values of minced beef containing 1% chitosan remained unchanged after 10 days of storage at 4 C (0.5 mg MDA/kg), whereas the values of control samples increased sharply. There was no significant difference between the pH values of the samples during the storage period (Table 2). pH of the control samples decreased from 6.74 to 6.41 whereas pH of the samples packed in antimicrobial bags ranged between 6.75 and 6.42 at the end of the storage period.
Fig. 3. Effect of antimicrobial packages on mold and yeasts (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
Fig. 4. Effects of different packages on TBARS value of chicken drumsticks.
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Table 2 Effect of different packages on the pH, color and hardness of chicken drumsticks. Property
Time (d)
Control ± SE
pH
0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00
6.740 6.720 6.560 6.600 6.670 6.410 76.603 73.683 74.817 75.523 78.877 75.787 3.637 4.653 4.923 3.270 3.147 4.203 11.220 8.483 12.093 4.593 9.363 7.870 2.708 6.494 4.197 3.804 2.690 2.461
L*-value
a*-value
b*-value
Hardness (N)
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.192aA 0.183aA 0.177aA 0.182aA 0.194aA 1.000abA 1.356bA 1.043abA 1.756abB 1.472bC 0.378abAB 0.267aA 0.271aA 0.321aA 0.762aA 0.537aB 0.967aA 0.270aA 0.498abAB 0.969aB 1.164bA 0.788aA 1.581abA 0.078aA 0.188bD 0.121cD 0.110dD 0.078aD 0.071bC
Chitosan ± SE 6.740 6.610 6.480 6.650 6.790 6.630 76.603 74.079 71.950 79.790 72.333 74.927 3.637 4.117 4.133 3.307 5.370 3.930 11.220 9.077 7.817 8.267 8.493 8.653 2.708 3.280 3.227 3.169 2.399 2.993
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
Nisin ± SE
0.1811aA 0.179aA 0.192aA 0.170aA 0.194aA 0.195aA 1.000abA 0.860acA 2.304cA 0.767bA 0.950acA 1.264acAB 0.267abA 0.460abA 0.909abA 0.150aA 0.251bA 0.869abA 0.270aA 0.704abA 0.690bA 0.421abAB 0.919abA 0.459abAB 0.078aA 0.095bA 0.093bcA 0.092bcA 0.069dA 0.086cA
6.740 6.730 6.740 6.520 6.650 6.420 76.603 75.297 72.420 76.210 77.420 74.293 3.637 4.013 4.673 5.103 3.440 4.983 11.220 8.813 7.870 13.183 7.927 13.673 2.708 4.583 3.745 2.201 2.037 3.322
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.183aA 0.191aA 0.178aA 0.190aA 0.181aA 1.000aA 1.430abcA 0.703bA 0.737aAB 0.318aBC 1.029abB 0.267abA 0.353abcA 0.260abcA 0.356bB 0.650aB 0.572bA 0.270abA 0.217bAB 0.745bA 0.265aC 0.164bA 0.226aC 0.078aA 0.132bB 0.108cB 0.064dB 0.059dB 0.096eB
PS ± SE 6.740 6.470 6.650 6.550 6.670 6.640 76.603 74.427 74.120 74.630 75.960 77.663 3.637 5.210 4.037 3.853 4.870 4.217 11.220 8.340 10.247 7.203 8.800 12.927 2.708 2.737 2.721 2.643 1.903 2.317
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
AgZeo ± SE 0.180aA 0.172aA 0.200aA 0.188aA 0.190aA 0.193aA 1.000abA 0.393abA 1.228aA 1.391abB 1.022abBC 0.584bB 0.267aA 0.209aA 0.358aA 0.094aAB 0.389aA 0.502aA 0.270abA 0.694cdAB 0.789acAB 0.337cAB 0.580acdA 0.841bC 0.078aA 0.079aC 0.079aC 0.134aC 0.055bB 0.067cC
6.740 6.670 6.800 6.730 6.750 6.750 76.603 75.910 73.177 76.307 74.810 73.863 3.637 3.597 4.177 4.023 4.957 5.130 11.220 6.993 10.200 9.647 8.390 11.573 2.708 3.221 3.937 3.349 2.340 2.778
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.182aA 0.191aA 0.180aA 0.190aA 0.192aA 1.000aA 0.684abA 0.895cA 0.452abAB 0.781abAB 0.833bA 0.267aA 0.215bA 0.362abcA 0.068abAB 0.303bA 0.471cA 0.270aA 0.579bB 0.583acAB 0.839abcBC 0.831bcA 0.784aBC 0.078aA 0.078bA 0.114cBD 0.097bA 0.068dA 0.080aA
Different capital letters indicate a statistical difference at a ¼ 0.05 level in each row. Different lower-case letters indicate statistical difference at a ¼ 0.05 level in each column.
3.2.3. Physical changes Color is an important quality attributes which affect the consumer acceptance and can give information about the food freshness. L-values of all samples are only slightly affected with the storage time. This finding is in agreement with results of Mexis, Chouliara, and Kontominas (2012) obtained for color parameters of antioxidant treated chicken meat. They reported that L*, a* and b* color parameter values of the samples remained unaffected (p > 0.05) as a function of storage time for samples containing citrus extract and/or an O2 absorber. There was no any significant change (p > 0.05) in the L-values of samples packed with different bags up to 2 days of storage period (Table 2). Sante and Lacourt (1994) suggested that application of vacuum protected the color of the samples due to a low rate of myoglobin oxidation. Similarly Chouliara et al. (2007) reported a decrease in L* values in chicken breast meat as a function of storage time in samples containing oregano oil. Ahn et al. (2000) observed large differences in all L*, a* and b* values between vacuum- and aerobic-packaging indicating that vacuum packaging was helpful in preserving meat color. A similar trend was observed for the change in Hunter a* and b* values of the all samples (Table 2) which is in agreement with results of Mexis et al. (2012). Use of different antimicrobial bags and storage time affected (p < 0.05) the hardness value (Table 2). The hardness values of the samples packed with chitosan and AgZeO were not different from each other (p > 0.05). The change in the hardness values was the highest (p < 0.05) for the samples packed with control bag and lowest in the samples packed with the PS bags. At the same time, the hardness value of samples in PS bags was not affected by storage time while that of the samples in the other bags was affected significantly. This could be due to low drip loss in drumstick packaged in PS compared to other samples. Use of antimicrobial packaging did not show any undesirable effect on the color
and texture of chicken drumsticks during the studied storage period. 4. Conclusions The results of this study indicated that the use of active bags and vacuum-sealing reduced the levels of APC, total coliform and mold and yeast count of chicken drumsticks when compared with the control. Of the antimicrobial agents examined, chitosan-containing film was the most effective film to increase shelf life and quality of the drumsticks. Efficiency of the active bags on the reduction of APC and total coliform was in the order of chitosan > nisin > AgZeo > PS, and on mold and yeasts reduction was in the order of chitosan > PS > nisin > AgZeo at 5 C for 6 days. Moreover, use of active bags protected the physical and chemical quality of chicken drumsticks compared to control samples. The results of the present study may be useful for the selection of the suitable antimicrobial packaging for chicken drumsticks. Chitosan or nisin can be suggested as potential use in antimicrobial packaging of foods. Addition of antimicrobials increase the cost of packaging materials but it is not more than 2%. These types of films have increased added value with benefits that would compensate for the increased cost. As a conclusion, shelf life and safety of chicken drumsticks could be increased by adjusting the level of active agents in films. Further studies are planned by use of these antimicrobial films to decrease addition of antimicrobial agents into foods and their possible migrations into foods. Acknowledgments We wish to thank to Industry Research and Development Supporting program of TUBITAK for financial support with the project number 3110257.
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Contents lists available at ScienceDirect
Food Control journal homepage: www.elsevier.com/locate/foodcont
Effect of antimicrobial packaging on physicochemical and microbial quality of chicken drumsticks dem Soysal a, *, Hüseyin Bozkurt a, Engin Dirican a, Mehmet Güçlü b, Çig Ebru Deniz Bozhüyük b, Ali Erdal Uslu b, Sevim Kaya a a b
Department of Food Engineering, Faculty of Engineering, University of Gaziantep, 27310 Gaziantep, Turkey NAKSAN Plastic Company, 1.Organized Industrial Zone, 83118 Street, Bas¸pınar, 27120, Gaziantep, Turkey
a r t i c l e i n f o
a b s t r a c t
Article history: Received 10 November 2014 Received in revised form 3 February 2015 Accepted 10 February 2015 Available online 18 February 2015
The effects of antimicrobial agents (nisin, chitosan, potassium sorbate (PS) or silver substituted zeolite (AgZeo)) incorporated into low density polyethylene (LDPE) on the physicochemical and microbial quality of chicken drumsticks stored at 5 C for 6 days were investigated. Active multilayer bags (LDPE/polyamide/LDPE-containing 2% antimicrobial agent) were used to pack chicken drumsticks under vacuum. Their efficiency in inhibiting total aerobic mesophilic bacteria (APC), total coliforms and total molds and yeasts were evaluated by comparison with control bag (LDPE-polyamide-LDPE). APC counts of samples packed in bags containing chitosan, nisin, AgZeo and PS were 1.03, 0.98, 0.51, and 0.17 log respectively were lower than those of samples packed in control bags. Samples packaged in active bags had lower microbial counts and thiobarbituric acid reactive substance (TBARS) values than those of samples packed in control bags. The pH, color and hardness of the samples were not affected significantly by use of different packaging films (p > 0.05). Therefore, the bag including chitosan or nisin can be suggested as a suitable packaging material for chicken drumstick to increase safety and quality. © 2015 Elsevier Ltd. All rights reserved.
Keywords: Active packaging Antimicrobial agent Chicken drumstick
1. Intoduction Active package changes the condition of the packed food to extend shelf-life or to improve safety or sensory properties, while maintaining the quality of packaged food (Ahvenainen, 2003; Hutton, 2003). Antimicrobial packaging appears to be one of the most promising applications of active food packaging technology (Joerger, 2007; Quintavalla & Vicini, 2002). The purpose of incorporating antimicrobials into film is to control spoilage as well as contamination with pathogens. Antimicrobial compounds incorporated into the packaging materials may be configured to be slowly released during food distribution and storage (Ishitani, 1995). As a result, microbial quality of packaged food products can be drastically improved and shelf life significantly extended. Commonly studied antimicrobial agents for active packaging are: nisin (Economou, Pournis, Ntzimani, & Savvaidis, 2009; Ercolini et al., 2010; La Storia et al., 2012; Pranoto, Rakshit, & Salokhe, 2005), chitosan (Chen, Weng, & Chen, 1999; Chounou
* Corresponding author. Tel.: þ90 342 317 23 07; fax: þ90 342 3601105. E-mail address: [email protected] (Ç. Soysal). http://dx.doi.org/10.1016/j.foodcont.2015.02.009 0956-7135/© 2015 Elsevier Ltd. All rights reserved.
et al., 2013; Quattara, Simard, Piette, Begin, & Holley, 2000), potassium sorbate (Han & Floros, 1997; Pranoto, et al., 2005; Weng & Hotchkiss, 1993), silver substituted zeolite (Brody, Stupinsky, & Kline, 2001; Ishitani, 1995), triclosan (Cutter, 1999; Vermeiren, Devlieghere, & Debvere, 2002), and essential oils (Hong, Park, & Kim, 2000; Oussalah, Caillet, Salmieri, Saucier, & Lacroix, 2004). Although these antimicrobial materials are active against several individual microorganisms under laboratory conditions, it has proven difficult to produce films that are sufficiently active in food systems (Balasubramanian, Rosenberg, Yam, & Chikindas, 2009). Multilayer packaging systems have been used to increase shelf life and safety of food products. Polymers can provide both improved barrier properties to both gas and water transfer and improved mechanical strength. Different polymers have different properties. For example, polyethylene, which is the most widely used polymer in food packaging, is a good barrier for water but a poor barrier for gases. Polyamide, however, provides a poor barrier for water but a good barrier for gases. So multilayer bags combining both possess higher barriers for both water and gases as well as providing increased mechanical strength. Incorporation of antimicrobial compounds into multilayer packaging polymers will be an alternative for limiting microbial growth and increasing shelf life of
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food products such as poultry meat. Poultry is widely consumed an important source of protein in human diet. Safety and quality of poultry meat are important issues for the food industry and also for consumers. Developing a method to increase shelf-life and overall safety/quality represents a major challenge for the poultry processing industry (Economou et al., 2009). Microbial contamination, which reduces the shelf-life of poultry meat and increases the risk of food borne illness, occurs mainly on the surface of poultry meat during slaughtering, processing, transportation and storage (Massoni, Morando, Vignolo, & Eisenberg, 2012). The objective of the current study was to determine the effect of different antimicrobial agents (chitosan, nisin, PS, AgZeo) on the microbial, physical and chemical characteristics of chicken drumsticks during refrigerated storage, compared with control multilayer bag without antimicrobial agent and having the same thicknesses.
2.3.1. Microbiological analysis Drumsticks were deboned under the aseptic conditions. About 25 g of drumstick sample was weighed aseptically, transferred to a flask containing 225 mL of sterile water and homogenized in a Waring Blender for 15 min at room temperature. For microbial enumerations, 0.2 mL samples of serial dilutions were spread on the surface of petri plates. APC was determined using Plate Count Agar after incubation at 37 ± 1 C for 2 days. Total mold and yeast counts were determined using Potato Dextrose Agar after incubation at 25 ± 1 C for 5 days. For total coliform counts, 0.2 mL sample was spread onto Violet Red Bile Agar and incubated at 37 ± 1 C for 2 days. Large colonies with purple haloes were counted. All plates were examined visually for typical colony types and morphological characteristics associated with each growth medium. Microbiological data are expressed as a logarithm of colony forming units per gram (log cfu/g).
2. Materials and methods
2.3.2. Chemical analysis The pH value was recorded using a pH meter. About 10 g of deboned chicken drumstick sample was homogenized in a Waring Blender with 90 mL of distilled water and the homogenate was used for pH determination. TBARS values of the samples were determined by a spectrophotometric method (Bozkurt, 2006). A 2-g sample was taken and TBARS were extracted twice with 10 mL of 0.4 M perchloric acid. The extracts were diluted to 25 mL with 0.4 M perchloric acid. The extracted solution was centrifuged for 5 min at 1790G. After centrifugation, 1 mL of supernatant was poured into a glass-stoppered test tube, 5 mL of TBA reagent was added and the mixture was heated in a boiling water-bath for 35 min. After cooling the tube in tap water, the absorbance of the sample was read against the appropriate blank at 538 nm. A calibration curve was prepared using malondialdehyde (MDA). TBARS values were expressed as mg of malondialdehyde/kg chicken sample.
2.1. Materials The polymers used in the present study were provided by NAKSAN Plastic Company (Gaziantep, Turkey). Nisin and chitosan were obtained from Handary SA, potassium sorbate and silver substituted zeolites were purchased from Aldrich Chemical Company, Inc., USA. Chicken drumsticks were obtained from a local slaughterhouse. 2.2. Film preparation The active and control bags were produced in NAKSAN Plastic Company. The active LDPE layer was obtained by mixing antimicrobial agents (2% of each chitosan, nisin preparation, PS, or AgZeo) with polyethylene pellets. Firstly, master batches were prepared by addition of 2% antimicrobial agent, 2% orevac as tie layer, 2% ethylene vinil acetate copolymer (EVA), and 94% polyethylene pellets, using a twin screw extruder. These master batches were used in production of active layers of the multilayer films. The active-multilayer (LDPE-polyamide-active LDPE) and control (LDPE -polyamide- LDPE) films having the same thicknesses (70 mm) were produced using a blown film extrusion process. Then films were sealed to prepare bags with 15 cm width and 25 cm length in sterile conditions. Oxygen transmission rates of the films were measured by the following procedure of ASTM D3985-05 (ASTM 2010) using an Ox-Tran Model 2/21 (Mocon, Inc., Minneapolis, Minn., U.S.A). The measurement conditions for oxygen permeability were 23 C, 100/0% relative humidity and 1 atm pressure. Permetran Model C 4/41 (Mocon, Inc., Minneapolis, Minn., U.S.A) was used to measure water vapor transmission rate of the films according to ASTM F1249-13 (ASTM 2013) at 23 C. 2.3. Packaging and storage of samples All chicken drumsticks were collected from the same batch of the slaughtered chickens, with all of them coming from the same farm. Chicken drumsticks were transported under chilled conditions to the laboratory and immediately packaged. The drumsticks were singly vacuum-packaged in active bags and control bags using a manual vacuum sealing machine, and stored at 5 C. For each packaging system, four samples were taken after 0, 1, 2, 3, 4, and 6 days of storage. The pH, color, TBARS values and microbiological analysis (total coliform, APC and mold and yeast counts) of the drumstick samples were followed after removal of the bags.
2.3.3. Physical analysis The changes in skin color (Hunter L*, a* and b*) of the drumsticks packed with active and control bags were determined from the same areas of the samples after removal of the bags, in all measurements. Chicken drumstick skin color was measured using a Hunter lab ColorFlex (A60-1010-615 Model colorimeter, Hunter lab, Reston, VA). The instrument was standardized each time with a white and black ceramic plates (Lo* ¼ 93.01, ao* ¼ 1.11 and bo* ¼ 1.27), and L*, a* and b* values of samples were recorded as average of four readings taken at same locations on each chicken drumstick. Hardness values of the chicken drumstick samples were determined using TA-XT2 Texture analyzer (Texture Technologies Corp., Scarsdale, NY/Stable Microsystems, Godalming, UK). Hardness of the samples was measured using a needle probe (1.8 mm in diameter) and recorded after penetration of the needle to a depth of 5 mm into sample at a speed of 1 mm/s. Data collection and calculation were done using Texture Expert Exceed Version 2 V3 (Stable Micro Systems, 1998). The procedure was repeated for two different samples and three times on each sample. The hardness is given as the mean of six measurements and expressed as Newton (N). All hardness values were determined from the same region of the samples. 2.4. Statistical analysis Analysis of Variance (ANOVA) was performed for each response (mold and yeast count, APC, total coliform count, pH, TBARS, L*, a*, b*, and hardness) to determine significant differences using the SPSS version 16.0 (SPSS Inc., Chicago, IL, USA). Duncan's multiple range tests were also carried out to distinguish examined groups.
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3.2.1. Microbiological quality APC, total coliform and mold and yeast counts are used by the poultry industry as general indicators of processing hygiene, storage quality and potential shelf-life (Capita, Alonso-Calleja, Garcia Fernandez, & Moreno, 2001; Alvarez-Astorga, Alonso-Calleja, ndez, 2002). Changes in APC (log Capita, Moreno, & Garcia-Ferna cfu/g) in chicken drumstick packaged with the active and control bags during 6 days of storage at 5 C are shown in Fig. 1. APC of all samples increased (P < 0.05) during storage and at the end of the storage period, APC of samples ranged from 4.70 to 5.73 log cfu/g. Similar results were reported by Kim and Marshall (1999) who found that APC in chicken leg increased to about 6 log cfu/g after 6 days of storage at 4 C. In our samples, the increase in APC in chitosan-incorporated packaging was about 0.5 log cfu/g whereas it was 1.53 log cfu/g in samples stored in control bags. The increases in APC were very low when compared with the data reported by Del Rio, Panizo-Mor an, Prieto, Alonso-Calleja, and Capita (2007) who reported a 6-log cfu/g increase in APC of chicken leg samples (from about 3.20 to 9.20 log cfu/g) after 5 days of storage at 3 C. Cox, Bailey, and Berrang (1998) reported that spoilage of poultry generally occurs when total aerobic mesophilic counts exceed 7 log cfu/g. In the present study, all drumsticks samples had lower than 6 log cfu/g after 6 days of storage.
Chicken drumsticks packaged with active bags had lower (P < 0.05) APC than the sample packed with control bags. At the 6th day of storage, APCs of samples packaged with nisin or chitosan bags were not statistically different (P > 0.05), but were lower (P < 0.05) APC than those of other samples. Similar results were also found by Siragusa, Cutter, and Willett (1999) and Park, Marsh, and Dawson (2010). Siragusa et al. (1999) used nisin incorporated low density polyethylene (LDPE) film for controlling food spoilage and enhancing product safety since nisin, a bacteriocin, inhibits Grampositive food borne pathogens and spoilage microorganisms. Park et al. (2010) observed that chitosan could be released from LDPE films to inhibit bacterial growth and that higher chitosan concentrations (1.4%) inhibited both gram positive and gram negative bacteria. The antibacterial property of chitosan is well known and is likely due to the interaction of the positively charged chitosan with negatively charged residues on the cell surface of many fungi and bacteria, causing extensive cell surface alterations and altering cell permeability. Changes in total coliform count in chicken drumstick packaged with control and active bags are shown in Fig. 2. Total coliform count increased (P < 0.05) during the 6 days of storage at 5 C from 3.19 log cfu/g to the counts ranging from 3.82 to 4.57 log cfu/g. Kim and Marshall (1999) observed that total coliform counts increased in chicken legs during storage and Del Rio et al. (2007) found that total coliform count increased from about 2.10 log cfu/g to about 7 log cfu/g during storage for 5 days at 3 C. In our study, the increase in total coliform count was lower (as about 1.4 log cfu/g) than that reported by Del Rio et al. (2007). Alvarez-Astorga et al. (2002) found that the contamination level of legs with coliforms was around 3.56 log cfu/g and reported that levels of coliforms in chicken skin varied between 2.7 and 4.9 log cfu/g. No reduction in Escherichia coli cells was observed in LDPE film containing chitosan polymer or oligomer (Hong et al., 2000) in a cast corn zein film containing lysozyme (Padgett, Han, & Dawson, 1998), and in an LDPE film coated with nisin (An, Kim, Lee, Paik, & Lee, 2000). However, in our study, use of active bags containing chitosan and nisin decreased (P < 0.05) total coliform count of chicken drumstick when compared with the control sample. The lowest (P < 0.05) total coliform count was observed in the chicken drumstick packed with chitosancontaining film whereas the highest counts (P < 0.05) was obtained with chicken drumstick packed in control bags. One-way ANOVA showed that mold and yeast counts were affected by both storage time and incorporation of antimicrobial
Fig. 1. Effects of antimicrobial packages on APC (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
Fig. 2. Effect of antimicrobial packages on total coliform counts (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
3. Results and discussions 3.1. Barrier properties of films Water vapor and oxygen transmission rates are important for controlling the barrier properties of films. Water and oxygen can accelerate the growth of microorganism and thus reduce the shelf life of foods. Water vapor transmission rates (g/m2$day) and oxygen transmission rates (cc/m2$day) of control bags were determined as 10.10 and 20.9, respectively. Addition of antimicrobial agents had no significant effect on the water vapor transmission rates of active bags which were found to be 9.55, 7.95, 8.15 and 7.85 g/m2$day for chitosan, nisin, PS and AgZeo added bags, respectively. However, oxygen transmission rates of active bags had significantly affected (P < 0.05) by the addition of chitosan (9.3), nisin (7.5), PS (7.0) and AgZeo (7.8 cc/m2$day). 3.2. Effect of antimicrobial packaging on quality of chicken drumsticks
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agents into the plastic bags. Mold and yeast count increased (P < 0.05) from 5.49 to levels ranging from 5.84 to 6.86 log cfu/g after 6 days of storage. As shown in Fig. 3, the drumsticks packed with active bags had lower (P < 0.05) mold and yeast counts than those packed in control bag. The rate of increase in mold and yeast counts during the storage periods was the highest in drumstick packed in control bags whereas it was the lowest in the samples packed in chitosan-containing bag. As a result, samples packaged with antimicrobial agents, especially with chitosan, nisin and PS, had low mold and yeast count throughout the 6 days of storage period. The efficiency of using chitosan-incorporated polyethylene extruded films in inhibiting radial growth of Aspergillus niger has been reported by Martínez-Camacho et al. (2013). There are conflicting results in the literature regarding the activity of PS, but one report suggested that the incorporation of PS into polyethylene films (0.4-mm thick) lowered the growth rate of mold and yeast (Han & Floros, 1997). Some authors stated that using low density polyethylene films (0.05-mm thick) containing 1.0% w/w sorbic acid were unable to suppress mold growth (Weng & Hotchkiss, 1993). Moreover, ethylene vinyl alcohol/linear low-density polyethylene (EVA/LLDPE) film (70 mm thick) impregnated with 5.0% w/w PS was unable to inhibit the growth of microorganisms on cheese (Devlieghere, Vermeiren, Bockstal, & Debevere, 2000). Results of the present study indicated that active bags have an inhibitory effect on microbial growth in chicken drumsticks when compared with the control. Table 1 shows the relative decrease in microorganisms (in log cfu/g) of samples packaged in active bags with respect to samples packaged in control bags at the end of the storage period. Chitosan was the most effective active bag with respect to APC, total coliform and mold and yeast counts. Nisin was also effective on inhibiting the APCs, total coliform and mold and yeasts. PS was found as effective on mold and yeast counts of chicken drumsticks. Although the microbial counts of samples packed with AgZeo were lower than the control samples, it was the least effective one around the studied antimicrobials.
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Table 1 Reduction in microbial counts (log cfu/g) of chicken drumstick in antimicrobial bags with respect to control bags at the end of storage period. Chitosan ± SE Nisin ± SE APC 1.03 ± 0.009 Mold and 1.02 ± 0.009 yeast Total 0.75 ± 0.104 coliform
PS ± SE
AgZeo ± SE
P-Value
0.98 ± 0.019 0.17 ± 0.060 0.51 ± 0.024 <0.01 0.87 ± 0.020 0.94 ± 0.004 0.25 ± 0.030 <0.01 0.68 ± 0.047 0.25 ± 0.163 0.28 ± 0.118 <0.05
Note: At the end of the storage period in counts of APC is 5.73 log cfu/g; mold and yeast is 6.86 log cfu/g; total coliform is 4.57 log cfu/g in control.
3.2.2. Chemical quality Lipid oxidation is a major contributor to quality deterioration process in muscle foods, resulting in a variety of breakdown products which produce undesirable off-odors and flavors. The initial TBARS value of the drumstick was 1.32 mg/kg sample. The suggested limit for TBARS value in meat products is 2 mg/kg above which rancid off-flavors become sensorially detectible (Byun et al.,
2003). After 2 days of storage, the TBARS values of the samples except chitosan were exceeded TBARS value of 2 mg/kg (Fig. 4). Although an increasing trend of TBARS values of the all samples was detected, there was no significant difference between those of the control samples and the samples packed with active bags, up to 2 days (p < 0.05). After 2 days of storage, TBARS values of all samples packaged with active bags were significantly lower (p < 0.05) than those of the samples packaged with control bags. Lower rate of lipid oxidation in samples packed with active bags compared to samples packed with control bags can also be related with the lower oxygen transmission rates of active bags compared to control bags. The susceptibility of drumstick meat towards oxidation has been explained by the high content of polyunsaturated fatty acids in this muscle (Chouliara, Karatapanis, Savvaidis, & Kontominas, 2007). Lowest lipid oxidation was observed for samples packed with chitosan incorporated bags, which might be helpful to limit lipid oxidation of the drumsticks during the storage period. The above results are in agreement with those of Georgantelis, Ambrosiadis, Katikou, Blekas, and Georgakis (2007) who reported a reduction in TBARS values for pork sausages by 73% for samples containing 1% chitosan (0.25 mg MDA/kg) as compared to control samples (0.96 mg MDA/kg after 10 days of storage at 4 C). Darmadji and Izumimoto (1994) reported that TBARS values of minced beef containing 1% chitosan remained unchanged after 10 days of storage at 4 C (0.5 mg MDA/kg), whereas the values of control samples increased sharply. There was no significant difference between the pH values of the samples during the storage period (Table 2). pH of the control samples decreased from 6.74 to 6.41 whereas pH of the samples packed in antimicrobial bags ranged between 6.75 and 6.42 at the end of the storage period.
Fig. 3. Effect of antimicrobial packages on mold and yeasts (log cfu/g) of chicken drumsticks stored at 5 C for 6 days.
Fig. 4. Effects of different packages on TBARS value of chicken drumsticks.
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Table 2 Effect of different packages on the pH, color and hardness of chicken drumsticks. Property
Time (d)
Control ± SE
pH
0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00 0.00 1.00 2.00 3.00 4.00 6.00
6.740 6.720 6.560 6.600 6.670 6.410 76.603 73.683 74.817 75.523 78.877 75.787 3.637 4.653 4.923 3.270 3.147 4.203 11.220 8.483 12.093 4.593 9.363 7.870 2.708 6.494 4.197 3.804 2.690 2.461
L*-value
a*-value
b*-value
Hardness (N)
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.192aA 0.183aA 0.177aA 0.182aA 0.194aA 1.000abA 1.356bA 1.043abA 1.756abB 1.472bC 0.378abAB 0.267aA 0.271aA 0.321aA 0.762aA 0.537aB 0.967aA 0.270aA 0.498abAB 0.969aB 1.164bA 0.788aA 1.581abA 0.078aA 0.188bD 0.121cD 0.110dD 0.078aD 0.071bC
Chitosan ± SE 6.740 6.610 6.480 6.650 6.790 6.630 76.603 74.079 71.950 79.790 72.333 74.927 3.637 4.117 4.133 3.307 5.370 3.930 11.220 9.077 7.817 8.267 8.493 8.653 2.708 3.280 3.227 3.169 2.399 2.993
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
Nisin ± SE
0.1811aA 0.179aA 0.192aA 0.170aA 0.194aA 0.195aA 1.000abA 0.860acA 2.304cA 0.767bA 0.950acA 1.264acAB 0.267abA 0.460abA 0.909abA 0.150aA 0.251bA 0.869abA 0.270aA 0.704abA 0.690bA 0.421abAB 0.919abA 0.459abAB 0.078aA 0.095bA 0.093bcA 0.092bcA 0.069dA 0.086cA
6.740 6.730 6.740 6.520 6.650 6.420 76.603 75.297 72.420 76.210 77.420 74.293 3.637 4.013 4.673 5.103 3.440 4.983 11.220 8.813 7.870 13.183 7.927 13.673 2.708 4.583 3.745 2.201 2.037 3.322
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.183aA 0.191aA 0.178aA 0.190aA 0.181aA 1.000aA 1.430abcA 0.703bA 0.737aAB 0.318aBC 1.029abB 0.267abA 0.353abcA 0.260abcA 0.356bB 0.650aB 0.572bA 0.270abA 0.217bAB 0.745bA 0.265aC 0.164bA 0.226aC 0.078aA 0.132bB 0.108cB 0.064dB 0.059dB 0.096eB
PS ± SE 6.740 6.470 6.650 6.550 6.670 6.640 76.603 74.427 74.120 74.630 75.960 77.663 3.637 5.210 4.037 3.853 4.870 4.217 11.220 8.340 10.247 7.203 8.800 12.927 2.708 2.737 2.721 2.643 1.903 2.317
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
AgZeo ± SE 0.180aA 0.172aA 0.200aA 0.188aA 0.190aA 0.193aA 1.000abA 0.393abA 1.228aA 1.391abB 1.022abBC 0.584bB 0.267aA 0.209aA 0.358aA 0.094aAB 0.389aA 0.502aA 0.270abA 0.694cdAB 0.789acAB 0.337cAB 0.580acdA 0.841bC 0.078aA 0.079aC 0.079aC 0.134aC 0.055bB 0.067cC
6.740 6.670 6.800 6.730 6.750 6.750 76.603 75.910 73.177 76.307 74.810 73.863 3.637 3.597 4.177 4.023 4.957 5.130 11.220 6.993 10.200 9.647 8.390 11.573 2.708 3.221 3.937 3.349 2.340 2.778
± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±
0.180aA 0.182aA 0.191aA 0.180aA 0.190aA 0.192aA 1.000aA 0.684abA 0.895cA 0.452abAB 0.781abAB 0.833bA 0.267aA 0.215bA 0.362abcA 0.068abAB 0.303bA 0.471cA 0.270aA 0.579bB 0.583acAB 0.839abcBC 0.831bcA 0.784aBC 0.078aA 0.078bA 0.114cBD 0.097bA 0.068dA 0.080aA
Different capital letters indicate a statistical difference at a ¼ 0.05 level in each row. Different lower-case letters indicate statistical difference at a ¼ 0.05 level in each column.
3.2.3. Physical changes Color is an important quality attributes which affect the consumer acceptance and can give information about the food freshness. L-values of all samples are only slightly affected with the storage time. This finding is in agreement with results of Mexis, Chouliara, and Kontominas (2012) obtained for color parameters of antioxidant treated chicken meat. They reported that L*, a* and b* color parameter values of the samples remained unaffected (p > 0.05) as a function of storage time for samples containing citrus extract and/or an O2 absorber. There was no any significant change (p > 0.05) in the L-values of samples packed with different bags up to 2 days of storage period (Table 2). Sante and Lacourt (1994) suggested that application of vacuum protected the color of the samples due to a low rate of myoglobin oxidation. Similarly Chouliara et al. (2007) reported a decrease in L* values in chicken breast meat as a function of storage time in samples containing oregano oil. Ahn et al. (2000) observed large differences in all L*, a* and b* values between vacuum- and aerobic-packaging indicating that vacuum packaging was helpful in preserving meat color. A similar trend was observed for the change in Hunter a* and b* values of the all samples (Table 2) which is in agreement with results of Mexis et al. (2012). Use of different antimicrobial bags and storage time affected (p < 0.05) the hardness value (Table 2). The hardness values of the samples packed with chitosan and AgZeO were not different from each other (p > 0.05). The change in the hardness values was the highest (p < 0.05) for the samples packed with control bag and lowest in the samples packed with the PS bags. At the same time, the hardness value of samples in PS bags was not affected by storage time while that of the samples in the other bags was affected significantly. This could be due to low drip loss in drumstick packaged in PS compared to other samples. Use of antimicrobial packaging did not show any undesirable effect on the color
and texture of chicken drumsticks during the studied storage period. 4. Conclusions The results of this study indicated that the use of active bags and vacuum-sealing reduced the levels of APC, total coliform and mold and yeast count of chicken drumsticks when compared with the control. Of the antimicrobial agents examined, chitosan-containing film was the most effective film to increase shelf life and quality of the drumsticks. Efficiency of the active bags on the reduction of APC and total coliform was in the order of chitosan > nisin > AgZeo > PS, and on mold and yeasts reduction was in the order of chitosan > PS > nisin > AgZeo at 5 C for 6 days. Moreover, use of active bags protected the physical and chemical quality of chicken drumsticks compared to control samples. The results of the present study may be useful for the selection of the suitable antimicrobial packaging for chicken drumsticks. Chitosan or nisin can be suggested as potential use in antimicrobial packaging of foods. Addition of antimicrobials increase the cost of packaging materials but it is not more than 2%. These types of films have increased added value with benefits that would compensate for the increased cost. As a conclusion, shelf life and safety of chicken drumsticks could be increased by adjusting the level of active agents in films. Further studies are planned by use of these antimicrobial films to decrease addition of antimicrobial agents into foods and their possible migrations into foods. Acknowledgments We wish to thank to Industry Research and Development Supporting program of TUBITAK for financial support with the project number 3110257.
Ç. Soysal et al. / Food Control 54 (2015) 294e299
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