Detection of antibiotic residues in broiler chicken meat in traditional markets in Surakarta City

This research aims to detect the presence of penicillin, tetracycline, aminoglycoside, and macrolide antibiotic residues in broiler chicken meat sold in traditional markets in Surakarta City. A total of 60 research samples were collected from traditional markets. Subsequently, the samples were examined at BBVet Wates, Yogyakarta, to identify the presence of antibiotic residues. The results showed that 16.67% of the chicken breast meat samples tested positive for penicillin and 10% showed traces of macrolide antibiotic residues. For chicken thigh meat, 63.33% tested positive for penicillin, while 3.33% contained macrolide antibiotic residues. However, no tetracycline and aminoglycoside antibiotic residues were detected in any of the meat samples. The presence of antibiotic residues can be attributed to several factors, including the excessive addition of antibiotics in feed, improper use of antibiotics in livestock. Based on the research results, it can be concluded that penicillin and macrolide antibiotic residues are still prevalent in broiler chicken meat sold in traditional markets in Surakarta. This shows there is a pressing need to strengthen guidance and supervision of antibiotic use regulations, as well as to monitor antibiotic residues in the meat before it reaches the market.


Introduction
The high consumption of meat necessitates careful consideration of the quality and safety of animal products.As indicated by [1], numerous issues persist concerning the quality and safety of animal products, with potential concerns arising from biological, chemical, and physical contaminants that may hazard human health.Among these contaminants, antibiotic residues stand out as a particular chemical concern found in animal products.
The consumption of animal-derived food containing antibiotic residues can result in various health disorders.Furthermore, the hazards associated with animal drug residues encompass both direct shortterm and long-term indirect consequences.These direct short-term effects include allergies, digestive disorders, skin disorders, anaphylaxis, and hypersensitivity, while long-term indirect effects involve microbiological resistance, neurological disorders, carcinogenicity, mutagenicity, teratogenicity, reproductive disorders, and tissue damage [2].
Cases of antibiotic residues have been reported in several regions across Indonesia.[1] reported the presence of antibiotic residues from penicillin, tetracycline, aminoglycoside, and macrolide groups in Bali, West Nusa Tenggara (NTB), and East Nusa Tenggara (NTT), with levels ranging from 0.3% to 9.15%.Moreover, [3] found that 8.33% of chicken meat samples from the Traditional Market in Yogyakarta contained oxytetracycline, while [4] found tetracycline residues in 7.14% of chicken meat samples sourced from various markets in Purwokerto.These results indicate the presence of antibiotic residues in poultry across different regions in Indonesia, associated with various antibiotic groups.Therefore, further research is warranted to investigate the presence of penicillin, tetracycline, aminoglycoside, and macrolide antibiotic residues in broiler chicken meat available in traditional markets in Surakarta.

Research tools
The tools employed for sample collection included a knife, cutting board, camera, and cooling box.To measure the size of the inhibition zone in antibiotic residue testing, a caliper was used.

Research materials
For this research, a total of 60 samples of broiler chicken meat were collected from traditional markets in Surakarta.The samples were carefully divided into 30 samples of chicken breast meat and 30 samples of chicken thigh meat.The antibiotic residue testing was carried out using the agar diffusion method.To evaluate the presence of specific antibiotic groups, bacterial strains were employed as references during the testing process.Specifically, Bacillus stearothermophilus (ATCC 7953) was used for penicillin, Bacillus cereus (ATCC 11778) for tetracycline, Bacillus subtilis (ATCC 6633) for aminoglycoside, and vegetative Kocuria rhizophila (Micrococcus luteus) (ATCC 9341) for macrolide groups.

Method
This research was conducted in several stages, including pre-survey, sample size calculation, collection of chicken meat samples, and examination of the samples at the Kesmavet in BBVet Wates.The sample in this research was chicken meat that was sold by sellers in traditional markets.The live animals were not used and did not violate animal welfare.Sample was taken by first asking the seller's permission.The following was an explanation of each stage.

2.3.1.
Pre-survey.The pre-survey stage was conducted to identify the traditional markets in Surakarta that sold broiler chicken meat.This was performed to determine whether the seller sold broiler chicken meat in each market.During this stage, the number of sellers in each market was also calculated, which served as the basis for determining the sample size.

Sample size calculation.
During the sample size calculation stage, the number of broiler chicken meat sellers to be included in the research on antibiotic residues was determined.The number of sellers used as samples was calculated using the detect disease formula.The determination of the sample size resulted from the calculation based on the formula for detecting the presence of disease [5]: The sample collection was conducted using a purposive sampling method.The sampling activity started with interviews with the sellers to record their identities.Each chicken meat sample was labeled with the name of their sellers and the origin of the market, and these samples were separated into breast and thigh parts.All samples were stored in a cool box and transported to the Kesmavet in Wates, Yogyakarta, for antibiotic residue testing in broiler chicken meat from the breast and thigh parts.

2.3.4.
Examination of samples at the Kesmavet in Wates, Yogyakarta.At the Kesmavet in Wates Veteriner Center, Yogyakarta, the collected samples were thoroughly examined to determine the presence of antibiotic residues (penicillin, tetracycline, aminoglycoside, and macrolide) in broiler chicken breast and thigh.The examination of samples strictly adhered to the guidelines outlined in [6], focusing on the Bioassay Screening Test Method for Antibiotic Residues in Meat, Eggs, and Milk.

Data analysis
Based on the examination conducted at the Kesmavet, Wates Veteriner Center, Yogyakarta, the data on antibiotic residues were analyzed descriptively.

Presence of antibiotic residues
The antibiotic residue test was conducted at the Kesmavet in BBVet Wates, Yogyakarta, following the [6] on the Bioassay Screening Test Method for Antibiotic Residues in Meat, Eggs, and Milk.The data on antibiotic residues in 60 broiler chicken meat from traditional markets in Surakarta City were presented in Table 1.Samples were obtained from the breast and thigh parts of broiler chicken meat because the administration of antibiotics in poultry was often performed intramuscularly (into the meat or muscle) in the thigh or breast parts [7].The results in Table 1 showed that antibiotic residues were detected in both the breast and thigh parts of the samples.Specifically, 26.67% of the samples tested positive for antibiotic residues in the breast part, while a higher percentage of 66.66% tested positive in the thigh part.This was evidenced based on the formation of inhibition zone diameters, which had a set limit for antibiotic residues at the rate of <13 mm.Therefore, when the diameter exceeded this zone rate, the sample was considered positive [6].Penicillin antibiotic residues were detected in breast samples with a percentage of 16.67% (5 out of 30), and macrolide antibiotics were identified in 10% of breast samples (3 out of 30).In the thigh samples, penicillin antibiotic residues were detected in a higher percentage of 63.33% (19 out of 30), with macrolide antibiotics being detected in a smaller proportion of 3.33% (1 out of 30).
All 60 samples analyzed in this research tested negative for tetracycline and aminoglycoside antibiotic residues, yielding a percentage of 0% (0/60).These aligned with the report of [8] and [9], where all samples from their respective investigation exhibited negative results for broiler chicken meat.[8] further emphasized that the absence of tetracycline antibiotic residues in broiler chicken meat could be attributed to the comprehensive understanding of farmers regarding proper antibiotic usage, including appropriate dosages, and strict adherence to withdrawal time.

Antibiotic residue risk factors
Various factors contributed to the presence of antibiotic residues.One primary factor was the excessive addition of antibiotics to animal feed, surpassing permissible limits.As outlined in the Regulation of the Minister of Agriculture No. 14 of 2017, certain antibiotics such as penicillin, aminoglycosides, macrolides, and tetracycline were classified as hard drugs due to their active ingredients, and their usage as additives in animal feed was strictly prohibited by the government.When these antibiotics were IOP Publishing doi:10.1088/1755-1315/1292/1/0120214 incorporated into the feed, they accumulated in the tissues of broiler chickens, exhibiting varying concentrations across the organs.Meanwhile, the majority of the parent compounds and their metabolites were eliminated from the body through urine and feces, others remained stored in the tissues, resulting in what was commonly known as residues [9].
The second contributing factor was the improper application of withdrawal time for antibiotics.[10] emphasized that withdrawal time was intended to allow for the complete elimination of the drug and its residues from the body of animals, depending on the specific type of drug used and the necessary duration.[11] further elaborated that failure to adhere to proper withdrawal protocols, particularly in chickens that were close to the harvesting and slaughter stage, could lead to the presence of antibiotic residues in the meat.In various cases, there was a short interval between the administration of the last antibiotic dose and the time of slaughtering.
The third factor was attributed to farmers not adhering to dosing and frequency guidelines recommended by veterinarians or those provided on the antibiotic packaging.Improper usage concerning dosage and frequency could lead to ineffective elimination of antibiotic residues from the body of animals [10].Using antibiotics in a manner inconsistent with the instructions on the product labels could consequently result in the presence of antibiotic residues in chicken meat.
The fourth factor was the contamination of feed with manure, which could occur due to the use of animal manure as fertilizer.In several cases, this manure was applied to fertilize feed crops, potentially leading to the presence of antibiotic residues in the animals that consumed the contaminated feed [12].

Antibiotic distribution in the chickens' body
The movement of drugs entering and leaving the body of animals involved several phases, including absorption, distribution, metabolism, and excretion.During the absorption phase, the compound entered the bloodstream, and this process was influenced by various factors, such as the characteristics of cell membranes, the physiological condition of the animal, and the properties and method of drug administration [13].The rate of dissolution also played a crucial role, where finer drug particles dissolved and reabsorbed faster [14].In the case of orally administered antibiotics, the drug particles were broken down into smaller molecules in the intestine and subsequently penetrated the walls of the small intestine.Furthermore, the drug was absorbed into the bloodstream through filtration, diffusion, or active transport.
The distribution process involved the transportation of drugs to all tissues and organs.However, drug distribution might not be uniform due to factors, including the blood-brain barrier, drug binding to blood proteins, or adipose tissue.For instance, antibiotics such as penicillin and tetracycline could cross these barriers with significant doses when administered intravenously [14].
The metabolism process occurred in the liver, which served as a significant site of accumulation for drugs.All drugs were transferred to the liver for metabolism.Within the liver, numerous binding sites were available for compounds that could not undergo detoxification or excretion.The elimination process occurred in the kidney.Drugs were excreted in the form of metabolites after undergoing biotransformation, and usually, only a small portion was excreted intact, such as penicillin and tetracycline.Additionally, this process occurred through the biliary system, where the drugs entered the small intestine and were eventually eliminated through feces.Antibiotics that were not entirely excreted would be transformed into residues.Certain antibiotic compounds entering the body of animals could not be entirely excreted, leading to their accumulation in tissues and subsequent formation of residues.

Impact of Antibiotic Residues
The presence of antibiotic residues in animal-derived food has significant implications, necessitating serious attention.Exceeding the Maximum Residue Limit (MRL) for antibiotic residues in animal products could lead to health impacts on consumers, including the following: Toxicological hazards pertained to contamination by various chemical substances in animal products that could affect consumers.These chemical substances have the potential to harm the survival of living organisms.Antibiotic residues posed toxicological hazards that encompassed mutagenicity, teratogenicity, and carcinogenicity [1].Mutagenicity referred to the ability to cause genetic changes, teratogenicity involved causing birth defects, and carcinogenicity induced the growth of cancer cells.
Microbiological hazards referred to contamination in animal products from harmful microorganisms that could jeopardize human health.These hazards included antibiotic resistance and disruption of normal gut flora [1].Antibiotic resistance occurred when bacteria developed the ability to protect against antibiotics, rendering the bacteria immune to their effects.[15] emphasized that other bacteria could thrive because susceptible ones have been inhibited or killed, allowing the remaining antibiotic-resistant bacteria to also conferred resistance to other bacteria.Moreover, antibiotics such as tetracycline could induce an imbalance in gut flora within the digestive tract, potentially leading to gastrointestinal tract irritation in certain individuals and causing changes in gut flora.These changes resulted in the development of infections and diarrhea.
Immunological hazards encompassed disturbances in all aspects of the immune system.According to [3], the immune system played a crucial role in identifying antigens or antibodies.These hazards could lead to allergic reactions or intolerances, which were hypersensitivity reactions.Such reactions might be manifested as hives on mucous membranes and skin, as well as rash and skin peeling.
Directly, antibiotics possessed toxic properties for humans, for instance, chloramphenicol was known to have severe side effects, including the suppression of bone marrow activity and disruption of red blood cell formation.This condition could lead to potentially fatal aplastic anemia.Most antibiotics used as therapeutic agents in domestic animals were also employed in humans.The toxicological hazards that occurred in humans due to antibiotic residues, particularly those from food products, were closely related to the dose and duration of exposure.In addition to its impact on health, antibiotic residues entailed economic consequences.The presence of antibiotic residues in animal-derived food could result in economic losses due to product rejection, particularly when these products were intended for export to countries with stringent and robust food safety systems.

Conclusion
The detection of antibiotic residues in broiler chicken meat in traditional markets in Surakarta city has indicated the presence of residues in both the breast and thigh parts.These detected residues belonged to the penicillin and macrolide groups.Based on these results, it became imperative to strengthen guidance and supervision of antibiotic usage regulations and closely monitor the presence of antibiotic residues in meat before it was introduced to the market.

Table 1 .
The result for antibiotic residues on broiler chicken meat