Czech J. Food Sci., 2026, 44(5):392-401 | DOI: 10.17221/154/2025-CJFS

Artisanal cheesemaking environment as a potential reservoir of antimicrobial-resistant EnterobacteriaceaeOriginal Paper

Zuzana Hanzelová1, Eva Dudriková ORCID...2, Franti¹ek Zigo ORCID...3, Viera Lovayová4
1 Department of Hygiene, Technology and Health Food Safety,University of Veterinary Medicine and Pharmacy, Ko¹ice, Slovak Republic
2 Center of Clinical and Preclinical Research MEDIPARK, Faculty of Medicine,Pavol Jozef ©afárik University in Ko¹ice, Ko¹ice, Slovak Republic
3 Department of Animal Nutrition and Husbandry, University of Veterinary Medicine and Pharmacy, Ko¹ice, Slovak Republic
4 Department of Medical and Clinical Microbiology, Faculty of Medicine, Pavol Jozef ©afárik University in Ko¹ice, Ko¹ice, Slovak Republic

The family Enterobacteriaceae is a group of undesirable bacteria in dairy plants. Their presence in cheesemaking could affect the food safety of the final products. The objective of this study was to detect, identify, and characterize representatives of the family Enterobacteriaceae in artisanal cheesemaking dairy plants in Slovakia. The susceptibility to different groups of antibiotics was tested using the disc diffusion method. Genes conferring resistance to beta-lactam antibiotics were detected by PCR with gene-specific primers. Of the 86 isolates, Enterobacter spp. was the most frequently identified genus (72.1%), followed by Klebsiella spp. (8.1%) and Leclercia spp. (8.1%). According to the CLSI criteria, the majority of isolates (77.9%) were resistant to cefazolin, followed by those resistant to cefoxitin (76.7%). According to the EUCAST criteria, the highest rates of resistance were to cefoxitin (84.9%) and ampicillin (66.3%). The presence of the blaTEM gene was confirmed in 42 (48.8%) Enterobacteriaceae isolates, the ampC gene in 9 (10.5%), and the blaCTX-M gene in 6 (6.9%). Our findings highlight the presence of Enterobacteriaceae in the artisanal cheesemaking environment, showing high levels of resistance to beta-lactam antibiotics and harboring the resistance genes.

Keywords: dairy plants; environmental samples; undesirable bacteria; MALDI-TOF MS; resistance genes

Received: September 11, 2025; Revised: June 29, 2026; Accepted: July 1, 2026; Prepublished online: October 7, 2026; Published: October 30, 2026  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Hanzelová Z, Dudriková E, Zigo F, Lovayová V. Artisanal cheesemaking environment as a potential reservoir of antimicrobial-resistant Enterobacteriaceae. Czech Journal of Food Sciences. 2026;44(5):392-401. doi: 10.17221/154/2025-CJFS.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Amador P., Fernandes R., Duarte I., Brito L, Prudêncio C. (2011): In vitro transference and molecular characterization of blaTEM genes in bacteria isolated from Portuguese ready-to-eat foods. World Journal of Microbiology Biotechnology, 27: 1775-1785. Go to original source...
    2. Aupaix A., Mzougui S., Soleimani R. (2024): Impact of susceptible, increased exposure, a new definition of the former intermediate susceptibility category introduced by the European Committee for Antimicrobial Susceptibility Testing on antimicrobial susceptibility testing evaluation. Clinical Microbiological Infection, 30: 1494-1496. Go to original source...
    3. Beyene A.M., Gizachew M., Yousef A.E., Haileyesus H., Abdelhamid A.G., Berju A., Tebeje M.M., Feleke T., Gelaw B. (2024): Multidrug-resistance and extended spectrum beta-lactamase-producing lactose fermenting Enterobacteriaceae in the human-dairy interface in northwest Ethiopia. PLoS ONE, 19: e0303872. Go to original source... Go to PubMed...
    4. Capita R., Castaño-Arriba A., Rodríguez-Melcón C., Igrejas G., Poeta P., Alonso-Calleja C. (2020): Diversity, antibiotic resistance, and biofilms-forming ability of Enterobacteria isolated from red meat and poultry preparations. Microorganisms, 8: 1226. Go to original source... Go to PubMed...
    5. Cherif-Antar A., Moussa-Boudjemâa B., Didouh N., Medjahdi K., Mayo B., Flórez A.B. (2016): Diversity and biofilm-forming capability of bacteria recovered from stainless steel pipes of a milk-processing dairy plant. Dairy Science and Technology, 96: 27-38. Go to original source...
    6. Clinical and Laboratory Standards Institute (2023): Performance Standards for Antimicrobial Susceptibility Testing. 33rd ed. CLSI supplement M100. Wayne, PA, CLSI.
    7. Crippa C., Pasquali F., Rodrigues C., De Cesare A., Lucchi A., Gambi L., Manfreda G., Brisse S. Palma F. (2023): Genomic features of Klebsiella isolates from artisanal ready-to-eat food production facilities. Scientific Reports, 13: 10957. Go to original source... Go to PubMed...
    8. Dey T.K., Lindahl J.F., Lundkvist Å., Grace D., Deka R.P., Shome R., Bandyopadhyay S., Goyal N.K., Sharma G., Shome B.R. (2023): Analyses of extended-spectrum-β-lactamase, metallo-β-lactamase, and AmpC-β-lactamase producing Enterobacteriaceae from the dairy value chain in India. Antibiotics, 12: 1449. Go to original source... Go to PubMed...
    9. Dokuta S., Zhang X., Jeeno P., Hongjaisee S., Yadoung S., Khamnoi P., Chuttong B., Kai Z., Hongsibsong S. (2025): ESBL-producing Enterobacterales in food and clinical samples: Antimicrobial resistance organisms and genes in Chiang Mai, Thailand. Scientific Reports, 15: 23886. Go to original source... Go to PubMed...
    10. Dudriková E., Záhumenská J., Výrostková J. (2022): Hygiena, technológia, kvalita a bezpeènos» mlieka a mlieènych výrobkov. 1st Ed. Ko¹ice, University of Veterinary Medicine and Pharmacy in Ko¹ice: 73-75. (in Slovak)
    11. European Committee on Antimicrobial Susceptibility Testing (2023): Breakpoint tables for interpretation of MICs and zone diameters. Version 13.0. Available at https://www.eucast.org/clinical_breakpoints/ (accessed July 31, 2025).
    12. EU/EEA (ESAC-Net) (2023): Annual Epidemiological Report 2023. Stockholm, ECDC. Available at https://www.ecdc.europa.eu/sites/default/files/documents/ESAC-Net_report-2023.pdf (accessed Sep 10, 2025).
    13. European Surveillance of Veterinary Antimicrobial Consumption (ESVAC) (2023): Sales Trends (mg/PCU) of Antibiotic VMPs in Slovakia for Food-Producing Animals. Amsterdam, EMA. Available at https://www.ema.europa.eu/en/veterinary-regulatory-overview/antimicrobial-resistance-veterinary-medicine/european-surveillance-veterinary-antimicrobial-consumption-esvac-2009-2023#annual-report-on-sales-of-veterinary-antimicrobial-medicinal-products-12821 (accessed July 31, 2025).
    14. Gelbíèová T., Koøená K., Pospí¹ilová-Hlucháòová L., Straková N., Karpí¹ková R. (2021): Dissemination and characteristics of Klebsiella spp. at processed cheese plant. Czech Journal of Food Science, 39: 113-121. Go to original source...
    15. Ke D., Picard F.J., Martineau F., Ménard C., Roy P.H., Ouellette M., Bergeron M.G. (1999): Development of a PCR assay for rapid detection of Enterococci. Journal of Clinical Microbiology, 37: 3497-3503. Go to original source...
    16. Kuang S., Mei F., Fu W., Zhong X., Bao C., Peng R., Yuan T. (2024): Identification and sterilization of β-lactamase-resistant Enterobacter spp. from fruits and raw vegetables. LWT - Food Science and Technology, 211: 116936. Go to original source...
    17. Larsson D.G.J., Flach C.F. (2021): Antibiotic resistance in the environment. Nature Reviews Microbiology, 20: 257-269. Go to original source... Go to PubMed...
    18. Mladenoviæ K.G., Grujoviæ M.®., Ki¹ M., Furmeg S., Tkalec V.J., Stefanoviæ O.D., Kociæ-Tanackov S.D. (2021): Enterobacteriaceae in food safety with an emphasis on raw milk and meat. Applied Microbiology and Biotechnology, 105: 8615-8627. Go to original source...
    19. Naghavi M., Vollset S.E., Ikuta K.S., Swetschinski L.R., Gray A.P., Wool E.E. et al. (2024): Global burden of bacterial antimicrobial resistance 1990-2021: A systematic analysis with forecasts to 2050. The Lancet, 404: 1199-1226. Go to original source... Go to PubMed...
    20. Nikoloudaki O., Junior W.J.F.L, Campanaro S., Di Cagno R., Gobbetti M. (2021): Role prediction of Gram-negative species in the resistome of raw cow's milk. International Journal of Food Microbiology, 340: 109045. Go to original source... Go to PubMed...
    21. Ntuli V., Njage P.M.K., Buys E.M. (2016): Characterization of Escherichia coli and other Enterobacteriaceae in producer-distributor bulk milk. Journal of Dairy Science, 99: 9534-9549. Go to original source... Go to PubMed...
    22. Pasquali F., Valero A., Possas A., Lucchi A., Crippa C., Gambi L., Manfreda G., De Cesare A. (2022): Occurrence of foodborne pathogens in Italian soft artisanal cheeses displaying different intra- and inter-batch variability of physicochemical and microbiological parameters. Frontiers in Microbiology, 13: 959648. Go to original source...
    23. Resendiz-Nava C.N., Silva-Rojas H.V., Rebollar-Alviter A., Rivera-Pastrana D.M., Mercado-Silva E.M., Nava G.M. (2022): A comprehensive evaluation of Enterobacteriaceae primer sets for analysis of host-associated microbiota. Pathogens, 11: 17. Go to original source... Go to PubMed...
    24. Yi S., Song H., Kim W.H., Lee S., Guk J.H., Woo J., Cho S. (2024): Dynamics of microbiota and antimicrobial resistance in on-farm dairy processing plants using metagenomic and culture-dependent approaches. International Journal of Food Microbiology, 417: 110704. Go to original source...

    This is an open access article distributed under the terms of the Attribution 4.0 International (CC BY 4.0.), which permits use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content