Assessment of Bacterial Contamination in Locally Sourced Chicken Meat from Markets Kendari City, IndonesiaAssessment of Bacterial Contamination in Locally Sourced Chicken Meat from Markets Kendari City, Indonesia

Ind. J. Anim. Agric. Sci., 6(1): 44-50, 2024 44

Assessment of Bacterial Contamination in Locally Sourced Chicken Meat from Markets Kendari City, Indonesia

Risma Wilasakti Risawa¹, Hamdan Has¹, Yamin Yaddi^1*, La Ode Sahaba¹, Fitria Dewi², Rina Astarika³, Denvy Meidian Daoed⁴, Deki Zulkarnain¹

1Departmentof Animal Science, Faculty of Animal Science, Halu Oleo University

2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Halu Oleo University

3Faculty of Science and Technology, Terbuka University -UPBJJ-Kendari

4Departmentof Animal Science, Faculty of Agriculture, Musamus University, Merauke

1,2Jl. H.E.A Mokodompit, Campus Hijau Bumi Tridharma, Anduonohu, Kendari City, Southeast Sulawesi, 93232, Indonesia

3JL. A.H. Nasution, Kendari City, Southeast Sulawesi, 93231, Indonesia

4Jl. Kamizaun Mopah Lama, Rimba Jaya, Merauke District, Merauke Regency, Papua, 99611, Indonesia

email: yaminyaddi@gmail.com

ABSTRACT

This research aims to ensure the safety of livestock-origin food products through the evaluation of microbial contamination potential in production-support facilities. The study was conducted by isolating and characterizing bacteria from poultry slaughter facilities in several traditional markets in Kendari City. Swab samples were taken from chicken holding pens in five traditional markets. The market locations were determined using purposive sampling, with the daily slaughter quantity as the main indicator. Isolation was performed by observing bacterial colony growth, and characterization was carried out by observing the morphology of bacterial colonies on specific differential media using Eosin Methylene Blue Agar (EMBA). Bacterial confirmation was done through microscopic observation based on the shape and morphology of bacteria in Gram staining. The results of this study indicate that bacteria can be isolated from all poultry slaughterhouses in traditional markets in Kendari City. Characterization results show that the bacteria were confirmed as Escherichia coli, Salmonella sp., and Pseudomonas sp. The low implementation of Good Hygiene Practices in production execution is a major supporting factor for the presence of bacteria as a source of contamination. Periodic supervision by relevant authorities is necessary as an effort to ensure the safety of livestock-origin food products.

Keywords: Bacteria, Chicken Holding Cage, Characterization INTRODUCTION

The decline in the quality of food ingredients is currently one of the challenges faced by consumers. This condition is caused by the high contamination of food sources by both chemical and biological materials, particularly in small-scale food processing. Biological contamination can occur through cross-contamination by workers or production equipment (Yaddi et al., 2023), while chemical contamination can happen during the maintenance period (Widayanti & Widwiastuti, 2018).

One highly vulnerable food ingredient to potential contamination is chicken meat. Its availability and affordable price make it a popular food choice among the public. Chicken meat is a source of protein and other non-essential nutrients needed by the body (Choi et al., 2023).

Ind. J. Anim. Agric. Sci.

e-ISSN: 2715-1697

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Ind. J. Anim. Agric. Sci., 6(1): 44-50, 2024 45 The nutritional content also provides an environment conducive for microbial growth. The major contaminants in chicken meat are often bacteria, especially aerobic bacteria. Various studies report that microbial contamination in chicken meat can come from opportunistic bacteria (Rizaldi et al., 2022) as well as pathogenic bacteria (Kholifah & Dharma, 2016).

The supply of chicken meat as a food ingredient in Kendari City still relies on traditional markets through Poultry Slaughterhouses (TPU). Before being slaughtered, broiler chickens are kept in specific pens for 1-7 days. The duration of containment depends on the demand for chicken meat. The slaughtering process at TPU that does not adhere to Good Hygiene Practice principles may lead to cross-contamination by bacteria in chicken meat through supporting equipment (Yaddi et al., 2023). Characterizing bacteria as contaminants in chicken meat is crucial for facilitating efforts to ensure the quality of food ingredients.

RESEARCH METHODS Research Materials

The research was conducted from January to April 2023. Bacterial isolation and characterization were performed at the Laboratory of Genetics and Animal Breeding, Faculty of Animal Science, while equipment preparation and media preparation were carried out at the Biomedical Laboratory, Faculty of Medicine, Universitas Halu Oleo, Kendari.

Materials used in the research included physiological NaCl, 70% alcohol, 96% alcohol, distilled water, aluminum foil, Eosin Methylene Blue Agar (EMBA), crystal violet, safranin, and lugol. The equipment utilized comprised test tubes, petri dishes, refrigerators, styrofoam, incubators, spatulas, forceps, microscopes, Erlenmeyer flasks, digital scales, analytical scales, autoclaves, laminar air flow, Bunsen burners, tweezers, tube racks, plastic gloves, and glass slides.

Research Procedure

The research implementation began by identifying the location of Poultry Slaughterhouses (TPU) in traditional markets as the sample source, with the daily slaughter quantity as the main consideration. After determining the sample source location, preparation and setup of equipment and materials were carried out. Samples were collected simultaneously within a specific timeframe using an organized working procedure. The transport medium for the samples used was physiological NaCl at a cold temperature.

Bacterial Isolation

The collected samples were isolated using the specific differential medium Eosin Methylene Blue Agar (EMBA). Using an inoculation loop, the samples were streaked in a zig- zag pattern on the surface of the medium. The inoculated medium was then incubated at 37°C for 24 - 48 hours. Positive results were indicated by the presence of bacterial colony growth on the surface of the EMBA medium.

Bacterial Characterization

Identification and characterization of bacteria were performed by distinguishing the shape and characteristics of bacterial colonies showing growth on the EMBA medium. The medium used is selectively differential, so the isolated bacterial colonies mostly originated

Ind. J. Anim. Agric. Sci., 6(1): 44-50, 2024 46 from Gram-negative bacterial groups. The differentiation of bacterial colonies showing growth was based on the bacteria's ability to ferment carbohydrates contained in the used medium.

Bacterial Confirmation

The characterized bacteria were then confirmed using Gram staining. Morphology and color of the bacteria served as the main indicators for confirming the results of bacterial characterization.

Data Analysis

The obtained data were presented in the form of figures and tables and then analyzed descriptively.

RESULTS AND DISCUSSION Bacterial Isolation

The results of bacterial isolation from chicken holding pens in the Kendari City traditional market showed bacterial growth (Table 1). Baruga Market is one of the sampling locations with the highest bacterial population. Meanwhile, the lowest population was shown in the Lapulu market and Korem market.

Table 1. Results of bacterial isolation on differential selective media

No Sample Origin EMBA

1 Baruga Market (100 %) 9^x /9^y

2 Panjang Market (90 %) 9^x /10^y

3 Andounohu Market (83 %) 5^x /6^y

4 Lapulu Market (70 %) 7^x /10^y

5 Korem Market (70 %) 7^x /10^y

Information: EMBA: Erosin Methylene Blue Agar, ^xnumber of samples with bacterial colony growth, ^ynumber of samples tested

Several studies have reported that various bacteria can be isolated from the environment and production facilities in Poultry Slaughterhouses (TPU) (Kartikasari et al., 2019), subsequently leading to contamination in chicken meat (Pratiwi et al., 2022). The contamination of chicken meat by microbes in the production/processing environment occurs due to the low implementation of sanitation practices (Rahmawati et al., 2018) and consistency in the application of biosecurity measures (Rudiyansyah et al., 2015). The presence of bacteria in the TPU environment can be attributed to contamination from chicken feces, harvesting baskets, and transporting vehicles.

The government has established types and threshold values for the presence of microbes in food products. This is done as an effort to ensure the quality of fresh or processed food ingredients. The presence of contaminant materials in food products can have direct or indirect consequences on consumers. Control at every stage of food processing is a key factor in achieving food safety. Implementation of control at all stages requires policies, awareness among food handlers, and supervision from business owners and the government (Nugraheni, 2013).

Ind. J. Anim. Agric. Sci., 6(1): 44-50, 2024 47 Karakterisasi dan Konfirmasi Bakteri

Characterization of bacteria on EMBA media shows the identification of three bacterial groups from all sample source locations (Table 2). These bacterial groups consist of Escherichia coli, Salmonella sp., and Pseudomonas sp. Characterization results reveal different distributions of bacteria from each Poultry Slaughterhouse (TPU). Bacterial groups are characterized based on the shape and color of colonies on the media surface (Figure 1a). The color formed on the media indicates the bacteria's ability to ferment carbohydrates (sucrose and/or lactose) as an energy source. Bacterial confirmation results indicate that all isolates belong to the Gram-negative bacterial group. Microscopic observations show rod-shaped bacteria with a red color (Figure 1b).

The high distribution of bacteria is suspected to be due to poor sanitation in the TPU environment, especially in holding pens, and the uncontrolled implementation of Good Manufacturing Practice. Bacterial contamination in the poultry slaughter environment has been widely reported, such as Escherichia coli as water contamination (Syarifuddin et al., 2020), Salmonella sp. on cutting equipment (Maulita et al., 2017), and Pseudomonas sp. in production wastewater (Aini et al., 2017). These bacteria are commonly found in poultry, both as normal flora (Escherichia coli and Pseudomonas sp.) and as pathogens (Salmonella sp.). However, under various conditions, these normal flora can infect poultry as hosts.

Table 2. Percentage of bacterial characterization results on EMBA media

No Sample Origin EMBA

Escherichia coli Salmonella sp. Pseudomonas sp.

1 Baruga Market (55 %) 5^x /9^y (22 %) 2^x /9^y (100 %) 9^x /9^y 2 Panjang Market (70 %) 7^x /10^y (50 %) 5^x /10^y (90 %) 9^x /10^y 3 Andounohu Market (83 %) 5^x /6^y (50 %) 3^x /6^y (83 %) 5^x /6^y 4 Lapulu Market (70 %) 7^x /10^y (60 %) 6^x /10^y (60 %) 6^x /10^y 5 Korem Market (70 %) 7^x /10^y (30 %) 3^x /10^y (60 %) 6^x /10^y Information: ^xnumber of samples with bacterial colony growth, ^ynumber of samples tested

The bacterial groups have established threshold values for chicken meat, such as Escherichia coli at 1 × 10^1 cfu/gram and Salmonella sp. negative/25 grams (SNI-01-7388- 2009). Standardization indicates that these bacterial types have the potential to act as pathogens for humans as consumers. The presence of these bacteria is a cause of foodborne zoonotic incidents (Abd El-Ghany, 2021).

Escherichia coli is a bacteria that inhabits the digestive tract of animals, including poultry. Some serotypes of Escherichia coli are reported to be pathogenic, possessing somatic antigens (O), capsule antigens (K), and flagellar antigens (H). The bacteria are often associated with occurrences of Avian Pathogenic Escherichia coli (APEC) causing colibacillosis in poultry (Ummamie et al., 2017). Escherichia coli infections are highly susceptible in young poultry, with clinical symptoms such as peritonitis, perihepatitis, pericarditis, and airsacculitis (Koutsianos et al., 2021). In humans, this bacterium is closely related to the occurrence of septicemia, diarrhea, and urinary tract infections (Bhavsar & Krilov, 2015).

Salmonella sp. is a pathogenic bacterium that affects both humans and animals. This bacterium is a key agent in the occurrence of zoonotic diseases and is closely associated with the occurrence of foodborne diseases. Salmonellosis in poultry is caused by Salmonella

Ind. J. Anim. Agric. Sci., 6(1): 44-50, 2024 48 pullorum, which affects all ages of poultry and leads to high mortality rates. In poultry, clinical symptoms can include chalky diarrhea and a decrease in egg production (Thaha, 2016).

Figure: Insert Image of Bacterial Colonies on EMBA Media (a). Characteristics show colonies of Salmonella sp. (green arrow), Pseudomonas sp. (white arrow), and Escherichia coli (yellow arrow). Confirmation using Gram staining (b) reveals rod-shaped bacteria in red color (blue arrow).

Pseudomonas sp. is a Gram-negative bacterial group that inhabits the digestive tract and is opportunistic in nature. This bacterium has been reported as a contaminant in chicken meat (Elbehiry et al., 2022) and eggs (Bengi et al., 2017). Some species within this bacterial group are zoonotic to humans and are closely associated with direct transmission. In poultry, Pseudomonas sp. infection leads to inflammation in the yolk sac and omphalitis (Eraky et al., 2020), while in humans, this bacterial group can cause inflammation in the respiratory tract.

CONCLUSION

The chicken holding pens at Poultry Slaughterhouses (TPU) in traditional markets represent a potential source of bacterial contamination in meat. The types of bacteria that can be isolated from these holding pens include Escherichia coli, Salmonella sp., and Pseudomonas sp. The lack of sanitation implementation in the TPU environment is a key factor predicting the high bacterial population in production and supporting facilities. Routine supervision is needed for business units involved in providing chicken meat as a food source.

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