Effect of Chitosan-Beeswax Edible Coating on Microbiological Profile of Chicken Thigh Meat at Freezer Storage
Salwa Haneefah Fauzia1), Dedi Fardiaz2), Laprianika Retha Hapita Sari3), Dewi Masyithoh4)
1) Research and Development, PT Kembang Joyo Sriwijaya, Jl. Raya Karangan, RT.12/RW.3, Jakaan, Bonowarih, Karang Ploso Subdistrict, Malang Regency, East Java 65152
2) Faculty of Dentistry, Muhammadiyah University, Jl. Mojopahit No. 666 B, Sidowayah, Celep, Sidoarjo Subdistrict, Sidoarjo Regency, East Java 61271
3) Faculty of Medicine, Airlangga University, Jl. Prof. DR. Moestopo No.47, Pacar Kembang, Tambaksari Subdistrict, Surabaya, East Java 60132
4) PT. Kembang Joyo Sriwijaya and Faculty of Animal Husbandry, University of Islam Malang, Dinoyo, Lowokwaru, Malang City, East Java 65144
*Corresponding Email: [email protected] Submitted 16 February 2024; Accepted 28 March 2024
ABSTRACT
This research examined the effect of chitosan and beeswax edible coating with different percentages on the microbiological value (aerobic bacteria, Staphylococcus aureus, Coliform, and Escherichia coli) of chicken thigh meat in two days of storage at freezer temperature (- 18oC to -20oC). The study used Completely Randomized Design with 4 treatments and 5 replications. Treatments included were A0 (control), A1 (chitosan 2%), A2 (chitosan 2% + beeswax 1%), and A3 (chitosan 2% + beeswax 2%). Variables observed were the number of aerobic bacteria, S. aureus, Coliform, and E. coli using 3MTM PetrifilmTM Plates. Results showed significant differences (p<0.05) in the number of aerobic bacteria colonies with the least colony on treatment A1. S. aureus and Coliform colonies showed significant differences (p<0.05) with the least colony on treatment A2 and A3, sequentially. There was no significant difference (p>0.05) in E. coli count due to no colonies detected on all treated samples.
However, compared to controlled samples, it was found to decrease. The decrease in colony numbers shows that edible coating treatments can be used to help preserve or extend the shelf life of chicken thigh meat.
Key words: Antimicrobial coating; chicken meat; food coating; frozen storage; petrifilm
INTRODUCTION
After COVID-19, Indonesians still tend to buy chicken meat online. However, due to meat’s perishable nature, preservation treatment is needed to prevent it from spoiling until it reaches the consumers.
Based on official sources from several major online stores in Indonesia, they generally deliver chicken meat with an estimated time of one hour to a maximum of two days to maintain its quality.
One method of preserving chicken meat that online sellers always use is freezing. The standard for freezing that is good to inhibit spoilage microbes’ growth on chicken meat ranges from -18oC to -20oC, and does not exceed -12oC (BPOM RI, 2021; Kaewthong et al., 2019; Triyannanto et al., 2021)
To optimize the preservation, sellers would also create an environment or treatment that limits air interaction and the loss or gain of moisture from the meat.
Because, air and high humidity can trigger the growth of spoilage microbes faster (Baltic et al., 2019). One of the modern innovations for this method is edible coating, a thin layer to coat foods that are safe to be eaten.
Examples of edible coating materials are chitosan and beeswax. Chitosan, a derivative of chitin made from shrimp and crab shells, is known to have antimicrobial properties (Zou et al., 2016; Verlee et al., 2017; Yan et al., 2021) and a relatively good gas barrier. However, it is permeable to water vapor and moisture (Souza et al., 2019; Naveed et al., 2019; Zhuang et al., 2020).On the other hand, materials made from lipids such as beeswax offer an effective barrier against water and moisture
due to their hydrophobic properties (Zhang et al., 2014; Vijayan et al., 2023). Also, according to Fratini et al. (2016) and Szulc et al. (2020), beeswax has been proven in many research to have antimicrobial effects such as on Staphylococcus and Escherichia bacteria. By combining these two materials, it is hoped to improve the quality of edible coating.
Observing the effect of edible coating can be done by calculating the microbe’s contamination, such as aerobic bacteria, Staphylococcus bacteria, and Coliform, specifically Escherichia coli (Baltić et al., 2015; Baltic et al., 2019). If it shows a decreasing number of colonies between untreated (control) samples and treated ones, then it can be said that edible coating treatments work positively in preserving the meat (Hermayasari, 2015; Apriliyani et al., 2020).
MATERIALS AND METHODS
Location and Time of Research
The research was conducted in June to December 2022 at the Microbiology Laboratory of Animal Science Faculty, University of Brawijaya, Malang.
Research Material
The materials were 20 chicken thigh carcasses bought at Best Meat butcher shop, Malang, chitosan powder (planet kimia), beeswax (belikimia), distilled water, acetic acid, and sodium chloride 0.9% for microbiological test dilution. The tools used were microbiological testing laboratory tools, a magnetic stirrer, 3MTM PetrifilmTM for testing Aerobic Bacteria Count (AC), S.
aureus count (STX), Coliform and E. coli count (EC), and a freezer for storage.
*Corresponding author:
Salwa Haneefah Fauzia
Email: [email protected]
Research and Development, PT Kembang Joyo Sriwijaya, Jl. Raya Karangan, RT.12/RW.3, Jakaan, Bonowarih, Karang Ploso Subdistrict, Malang Regency, East Java 65152
How to cite:
Fauzia, S. H., Fardiaz, D., Sari, L. R. H., &
Masyithoh, D. (2024). Effect of Chitosan-Beeswax Edible Coating on Microbiological Profile of Chicken Thigh Meat at Freezer Storage. Jurnal Ilmu dan Teknologi Hasil Ternak, 19 (1), 63-68
Research Method
This research used a Completely Randomized Design (CRD) with 4 treatments and 5 replications. The treatments applied to chicken thigh meat samples were:
A0 = without any treatment (control) A1 = chitosan 2% edible coating, A2 = chitosan 2 % + beeswax 1%, A3 = chitosan 2 % + beeswax 2%
Samples were then stored in a freezer temperature of -18oC to -20oC for two days.
Chitosan-beeswax edible coating making are based on the method by Eldaly, et al.
(2018), Foo, et al. (2018), and Yolanda, et al. (2021) with modifications.
The method for testing and counting bacteria refers to the Official Interpretation Guide from 3MTM PetrifilmTM plates (2017), SNI 2897:2008 regarding testing for microbial contamination using the total plate count (TPC) method on meat (for aerobic bacteria), SNI 2897:2008 regarding methods testing for microbial contamination in meat, eggs, milk and processed products, and SNI 2332.9:2011 concerning microbiological testing methods – Part 9: determination of Staphylococcus aureus in fishery products
(for S. aureus) and AOAC Official Method 998.08 concerning confirmation of the number of E. coli in poultry, meat, and seafood.
Research Variable
Variables observed were the colony number of aerobic bacteria, S. aureus, Coliform and E. coli according to the official counting procedures in 3MTM PetrifilmTM Plates Interpretation Guide (2017).
Data Analysis
The aerobic bacteria was analyzed using analysis of variance (ANOVA) and followed by Duncan’s Multiple Range Test (DMRT). The number of S. aureus, Coliform, and E. coli were analyzed by Kruskal-Wallis test followed by the Mann- Whitney Test to identify the differences more specifically (Weaver et al., 2017).
RESULTS AND DISSCUSION
Aerobic Bacterial Number
The results showed that the edible coating was significantly decreased the aerobic bacteria (P<0.05) (Table 1). The lowest of aerobic bacteria number was found in A1, followed by A2, A3, and A0.
Table 1. Results of bacteria numbers
Treatment Aerobic bacteria (log cfu/g)
S.aureus (log cfu/g)
Coliform (log cfu/g)
E.coli (log cfu/g)
A0 5.93d±0.13 3.37±0.29 3.34±0.39 1.28±1.76
A1 4.14a±0.27 2.25±1.28 1.26±1.73 Not identified
A2 4.63b±0.14 1.14±1.56 0.74±1.65 Not identified
A3 5.18c±0.10 3.24±0.22 0.60±1.34 Not identified
Remarks : *Mean values within a column followed by the different letters are significantly different at p < 0.05 according to Duncan’s Multiple Range Test.
This is in line with the results in the use of chitosan to reduce the number of aerobic bacteria numbers, as by Eldaly et al.
(2018) on chicken fillets during cold storage and Abdel-Naeem et al. (2021) on drumsticks in freezer storage. Furthermore, Apriliyani et al. (2021) also reported a decrease in bacteria count in chicken meat applied with chitosan-casein edible coating mixed with 0.05% liquid beeswax. These results support the statement that chitosan
has antimicrobial or antibacterial properties, due to electrostatic interactions between the polycationic structure of chitosan and the anionic groups on bacterial cell surfaces, resulting in bacterial death (Younes et al., 2015; Zou et al., 2016; Verlee et al., 2017).
Additionally, chitosan coatings serve as a good barrier against oxygen transfer, inhibiting aerobic bacteria growth (Liu et al., 2019). Whereas pure lipids such as beeswax, are an excellent moisture, vapor, and oxygen
barrier, thus helping in inhibiting microbial growth, including aerobic bacteria
(Sanchez-Ortega et al., 2014; Trevisani et al., 2017).
Staphylococcus aureus Number
The results showed that the edible coating was significantly decreased the S.
aureus bacteria (P<0.05) (Table 1). The lowest of S. aureus number was found in A2, followed by A1, A3, and A0. Chitosan has been proven to be effective in inhibiting the growth of S. aureus in chicken meat, either by itself or mixed with other coating materials and oils (Eldaly et al., 2018;
Apriliyani et al., 2020; Hadidi et al., 2020).
Chitosan consists of glucosamine and N- acetyl glucosamine units connected by β (1- 4) glucoside bonds, which this polymer has antimicrobial activity against Staphylococcus aureus (Ngo et al., 2015;
Muñoz-Tebar et al., 2023). Whereas beeswax according to Prestianti et al. (2018) has good antibacterial activity against S.
aureus when extracted with methanol and ethyl acetate solvents.
Results of Treatments on Coliform The results showed that the edible coating was significantly decreased the Coliform bacteria (P<0.05) (Table 1) with the lowest number was found in A3, followed by A2, A1, and A0.
The successful use of chitosan has been proven to inhibit the growth of coliform bacteria in chicken meat with the addition of Trachyspermum ammi essential oil (Karimnezhad et al., 2017), mixed with rosemary essential oil (Souza et al., 2019), and mixed with Artemisia fragrans essential oil in refrigerator storage (Yaghoubi et al., 2021).
Escherichia coli Number
The results showed that the edible coating did not significantly decrease E. coli bacteria (P>0.05) (Table 1) with the lowest number was found in A3, followed by A2, A1, and A0. On this research, Escherichia coli colonies on samples treated with edible coatings were not identified. According to the 3MTM PetrifilmTM EC Plate
Interpretation Guide (2017), it is indeed possible for E. coli not to be detected, by the absence of blue colonies with gas on the plate, which also could mean there are no E.
coli found in the samples. However, by observing the mean value data, there were still E. coli colonies identified on control samples, which could probably show an effect from edible coatings that serve as a protective barrier against E.coli.Research by Raphaël and Meimandipour (2017) and Souza, et al. (2019) showed the antibacterial activity of chitosan mixed with essential oils against Escherichia coli. This is because the positively charged chitosan supplemented with essential oils creates a semi-permeable barrier that can reduce respiration and delay microbial growth. Research by Prestianti, et al. (2018) showed that beeswax extract dissolved in methanol and n-hexane showed good antibacterial activity.
CONCLUSION
Chitosan-beeswax edible coating can be used and researched further to extend the life of chicken meat that is stored frozen, as seen from the results of reducing the number of colonies of aerobic bacteria, S. aureus, and coliform. However, this edible coating has not been proven to help prevent the growth of e.coli contamination in frozen chicken meat.
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