Detection of antibiotic residues of Tetracycline, Oxytetracycline, and Chlortetracycline in animals' raw milk in Hamedan province, Iran in 2022 using the Four-Plate Test, ELISA, and HPLC techniques

Document Type : Original Articles

Authors

1 Hamadan University of Medical Sciences

2 Nutrition Health Research Center, Hamadan University of Medical Sciences, Hamadan, Iran.

3 Infectious Diseases Research Center, Hamadan University of Medical Sciences, Hamadan, Iran.

4 Department of Epidemiology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

5 Iranian Veterinary Organization, Hamedan, Iran.

10.32592/ARI.2025.80.3.803

Abstract

Antibiotic resistance has become a health concern as it is associated with the death of numerous people worldwide. Milk safety is one factor that guarantees the quality of dairy products. This study was designed to determine Tetracycline (TC), Oxytetracycline (OTC), and Chlortetracycline (CTC) residues in raw milk of animals from Hamedan, Iran using a Four-plate test (FPT), enzyme-linked immunosorbent assay (ELISA), and high-performance liquid chromatography (HPLC) techniques. Cross-sectionally over two years, 246 unprocessed raw milk samples were taken from dairy farms and milk collection centres of different regions of Hamedan, the western part of Iran. FPT was the first tool for screening the presence of antibiotics. Then, the positive samples were analyzed for antibiotic residue using ELISA. Finally, the HPLC method was applied to determine the type and amount of Tetracycline residues. In the primary evaluation, forty-seven (19.11 %) samples were positive for antimicrobial residues using FPT. ELISA analysis indicated that 29.79 % (14/47) of samples had a level of TCs higher than the maximum residue limit (MRL) suggested by EU (100 µg/L). The average TCs residue in positive samples was calculated 98.43±6.86 µg/L. The lowest and highest levels were 100.59 µg/L and 129.56 µg/L, respectively. Finally, the average TCs was calculated 105.73±7.25 µg/L (TC=100.67, OTC=103.38, and CTC=107.11 μg/L) using HPLC. The detection of antibiotic residues in animal products highlights the need for monitoring such residues in milk and other animal-origin food products. Training farmers for the correct use of drugs, especially antibiotics, is recommended. A comprehensive protocol for regularly evaluating livestock products is necessary to prevent high-contamination products from entering the production cycle.

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