Skip to main content
SpringerLink
Log in

Development and Evaluation of a Preliminary Screening Assay for Antibiotic Residues in Meat

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

A preliminary screening assay based on a microbial chromogenic reaction was developed to detect common antibiotic residues in meat rapidly. The assay comprised two bioassays: one for Escherichia coli and another for Geobacillus stearothermophilus. The assay was optimized and evaluated for the simultaneous screening of 30 antibiotics from five common antibiotic classes (tetracyclines, aminoglycosides, macrolides, β-lactams, and quinolones) found in meat. Extraction using phosphate-acetonitrile buffer (pH 7.2) and a delipidating treatment using n-hexane resulted in a high extraction efficacy for the five antibiotics, without affecting the microbial color reaction. A carrier, polyvinyl alcohol (0.1 g/mL); a cross-linking agent, boric acid-sodium tetraborate solution (pH 5.5); and a bacterial suspension with an initial optical density of 1.0 were the optimal embedding conditions for stability, microbial activity, and chromogenic efficiency. The assay exhibited a 6-month shelf life, with detection limits of 40–60, 60–140, 60–100, 20–40, and 40–180 μg/kg for tetracyclines, aminoglycosides, macrolides, β-lactams, and quinolones, respectively, which met the European Commission (37/2010) requirements for antibiotic residue limits. Our assay results were confirmed using LC-MS/MS with 160 samples, revealing a good correlation. This study demonstrates a reliable, easy-to-use, and economical method for preliminary screening of antibiotic residues in meat. This method may find an immediate application in food safety and general testing laboratories.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data Availability

The data produced and/or analyzed in the current study are available from the corresponding author on reasonable request.

References

  1. Zhu, H. (2016). Hazards of antibiotic residues in animal-derived foods and detection methods. Industrial & Science Tribune, 15, 64–65.

    Google Scholar 

  2. Jank, L., Martins, M. T., Arsand, J. B., Motta, T. M. C., Feijó, T. C., dos Santos Castilhos, T., & Pizzolato, T. M. (2017). Liquid chromatography–tandem mass spectrometry multiclass method for 46 antibiotics residues in milk and meat: development and validation. Food Analytical Methods, 10(7), 2152–2164. https://doi.org/10.1007/s12161-016-0755-4.

    Article  Google Scholar 

  3. Li, B., & Ji, Z. (2019). Main detection techniques and coping strategies for antibiotic residues in animal products. Poultry Science, 2, 53–55.

    Google Scholar 

  4. Zheng, Y., Fan, L., & Dong, Y. (2019). Detection technology for veterinary antibiotic residue in livestock and poultry products: a review. Quality and Safety of Agro-products, 3, 65–69.

    Google Scholar 

  5. Chen, X., & Ye, N. (2016). Graphene oxide-reinforced hollow fiber solid-phase microextraction coupled with high-performance liquid chromatography for the determination of cephalosporins in milk samples. Food Analytical Methods, 9(9), 2452–2462. https://doi.org/10.1007/s12161-016-0435-4.

    Article  Google Scholar 

  6. Ramatla, T., Ngoma, L., Adetunji, M., & Mwanza, M. (2017). Evaluation of antibiotic residues in raw meat using different analytical methods. Antibiotics, 6(4), 34. https://doi.org/10.3390/antibiotics6040034.

    Article  CAS  PubMed Central  Google Scholar 

  7. Hennart, S. L. A., & Faragher, J. (2012). Validation of the Delvotest® SP NT. Journal of AOAC International, 95(6), 1807–1815. https://doi.org/10.5740/jaoacint.12-232.

    Article  CAS  PubMed  Google Scholar 

  8. Nagel, O., Molina, M., & Althaus, R. (2013). Use of chemometric techniques to design a microbiological method for sulfonamide detection in milk. Czechoslovak Journal of Food Sciences, 31(6), 627–632.

    Article  Google Scholar 

  9. Karageorgou, E., Christoforidou, S., Ioannidou, M., Psomas, E., & Samouris, G. (2018). Detection of β-lactams and chloramphenicol residues in raw milk-development and application of an HPLC-DAD method in comparison with microbial inhibition assays. Foods, 7(6), 82. https://doi.org/10.3390/foods7060082.

    Article  CAS  PubMed Central  Google Scholar 

  10. Wu, Q., Peng, D., Liu, Q., Shabbir, M. A. B., Sajid, A., Liu, Z., & Yuan, Z. (2019). A novel microbiological method in microtiter plates for screening seven kinds of widely used antibiotics residues in milk, chicken egg and honey. Frontiers in Microbiology, 10, 436. https://doi.org/10.3389/fmicb.2019.00436.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Zhu, J. (2018). Application and analysis of antibiotic in milk by microbial chromogenic assay. China Food Safety Magazine, 21, 66–67.

    Google Scholar 

  12. Lee, D. D., Lee, E. Y., Jeong, S. H., & Chang, C. L. (2007). Evaluation of a colorimetric broth microdilution method for antimicrobial susceptibility testing using 2,3,5-triphenyltetrazolium chloride. Korean Journal of Clinical Microbiology, 10(1), 49–53.

    Google Scholar 

  13. Tumini, M., Nagel, O. G., Molina, P., & Althaus, R. L. (2016). Novel bioassay using Bacillus megaterium to detect tetracycline in milk. Revista Argentina de Microbiología, 48(2), 143–146. https://doi.org/10.1016/j.ram.2016.02.001.

    Article  PubMed  Google Scholar 

  14. Huang, X., Zheng, J., Wu, Q., Chen, B., & Tang, M. (2008). Study on MIA method for determination of antibiotics residues in food. Food Science, 28(8), 418–421.

    Google Scholar 

  15. Hu, K., Li, Y., Zhu, Z., Huan, X., Wang, M., Fang, S., & Yang, X. (2009). A rapid method to determine the residues of enrofloxacin in fishery products by microbiological chromotest. Journal of Shanghai Ocean University, 18(4), 472–478.

    Google Scholar 

  16. Wei, G. (2018). Residue analysis of β-lactams antibiotics drugs in animal derived food by microbiological inhibition assay. The Light & Textile Industries of Fujian, 4, 40–42.

    Google Scholar 

  17. Wang, L., Si, X., & Tang, H. (2019). Residue detection of seven antibiotics in livestock and poultry meat. Chinese Journal of Analytical Chemistry, 15, 1–5.

    Google Scholar 

  18. Jammoul, A., & El Darra, N. (2019). Evaluation of antibiotics residues in chicken meat samples in Lebanon. Antibiotics, 8(2), 69. https://doi.org/10.3390/antibiotics8020069.

    Article  CAS  PubMed Central  Google Scholar 

  19. Benavides, S., Cortés, P., Parada, J., & Franco, W. (2016). Development of alginate microspheres containing thyme essential oil using ionic gelation. Food Chemistry, 204, 77–83. https://doi.org/10.1016/j.foodchem.2016.02.104.

    Article  CAS  PubMed  Google Scholar 

  20. Lui, Y., Miao, Y., & Wang, J. (2016). PVA immobilized microorganism method and application. Science and Technology Review, 34, 56–61.

    Google Scholar 

  21. Deng, Y., Huo, A., & Li, S. (2018). Process optimization, characterization, and functional properties analysis of Bifidobacterium composite microcapsules. Science and Technology of Food Industry, 10, 23–24.

    Google Scholar 

  22. Index of Veterinary Drugs | CODEX ALIMENTARIUS FAO-WHO. (n.d.). Available from: http://www.fao.org/fao-who-codexalimentarius/codex-texts/dbs/vetdrugs/veterinary-drugs/en/. Accessed June 24, 2020.

  23. Tumini, M., Nagel, O. G., & Althaus, R. L. (2019). Five-assay microbiological system for the screening of antibiotic residues. Revista Argentina de Microbiología, 51(4), 345–353. https://doi.org/10.1016/j.ram.2019.01.002.

    Article  PubMed  Google Scholar 

  24. Sanz, D., Mata, L., Condón, S., Sanz, M. Á., & Razquin, P. (2011). Performance of a new microbial test for quinolone residues in muscle. Food Analytical Methods, 4(2), 212–220. https://doi.org/10.1007/s12161-010-9151-7.

    Article  Google Scholar 

  25. Mata, L., Sanz, D., & Razquin, P. (2014). Validation of the Explorer ® 2.0 test coupled to e-Reader ® for the screening of antimicrobials in muscle from different animal species. Food Additives and Contaminants A, 31(9), 1496–1505. https://doi.org/10.1080/19440049.2014.934303.

    Article  CAS  Google Scholar 

  26. Schneider, M. J., & Lehotay, S. J. (2008). A comparison of the FAST, Premi® and KISTM tests for screening antibiotic residues in beef kidney juice and serum. Analytical and Bioanalytical Chemistry, 390(7), 1775–1779. https://doi.org/10.1007/s00216-008-1918-y.

    Article  CAS  PubMed  Google Scholar 

  27. Wang, Y., Li, Y., & Yan, G. (2016). Advances in antibiotic residues pre-treatment and determination method. Shandong Industrial Technology, 20, 292–293.

    Google Scholar 

  28. Zhang, P., Shen, Z., & Zhang, Y. (2018). Drug resistance characteristics and epidemiological analysis of 8 isolates of NDM-1-producing Enterobacteriaceae. Laboratory Medicine, 33, 616–621.

    Google Scholar 

  29. Bohn, T., Pellet, T., Boscher, A., & Hoffmann, L. (2013). Developing a microbiological growth inhibition screening assay for the detection of 27 veterinary drugs from 13 different classes in animal feeding stuffs. Food Additives and Contaminants A, 30(11), 1870–1887. https://doi.org/10.1080/19440049.2013.832400.

    Article  CAS  Google Scholar 

  30. Martins, E., Poncelet, D., & Renard, D. (2017). A novel method of oil encapsulation in core-shell alginate microcapsules by dispersion-inverse gelation technique. Reactive and Functional Polymers, 114, 49–57. https://doi.org/10.1016/j.reactfunctpolym.2017.03.006.

    Article  CAS  Google Scholar 

  31. Lin, J., & Jin, H. (2015). Experiment on rural domestic wastewater treatment by PVA immobilized microorganism and bamboo charcoal. Guangzhou Chemical Industry, 40, 104–105.

    Google Scholar 

Download references

Funding

This work was supported by the National Key R&D Program of China (grant number 2018YFC1603400).

Author information

Authors and Affiliations

  1. China Meat Food Research Center, Beijing Academy of Food Sciences, Beijing, 100068, China

    Wei Fan, Xiao-yue Gao, Ming-wu Zang, He-nan Li, Wen-ping Guo, Ying-ying Li & Shou-wei Wang

Authors
  1. Wei Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiao-yue Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ming-wu Zang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. He-nan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wen-ping Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ying-ying Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shou-wei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

The study was conceived and designed by Shou-wei Wang, Wen-ping Guo, and Ying-ying Li. The first draft of the manuscript was written by Wei Fan. Statistical analyses were performed by Wei Fan and Xiao-yue Gao. Sections of the manuscript were written by Wei Fan, He-nan Li, and Ming-wu Zang. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Shou-wei Wang.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethics Approval and Informed Consent

Not applicable.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Code Availability

Not applicable.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fan, W., Gao, Xy., Zang, Mw. et al. Development and Evaluation of a Preliminary Screening Assay for Antibiotic Residues in Meat. Appl Biochem Biotechnol 193, 1129–1146 (2021). https://doi.org/10.1007/s12010-020-03462-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-020-03462-z

Keywords

Search

Navigation