journal article·2026-04-10Open access

Raw Milk Cheeses as Reservoirs of Antimicrobial-Resistant Bacteria: A Comparative Study of Goat and Sheep Milk Products

Kimia Dalvand, Katarzyna Ratajczak, Paweł Cyplik, Jakub Czarny, Agnieszka Piotrowska-Cyplik

Applied Sciences· Volume 16· Issue 8

Abstract

This study investigated the microbiological composition and antimicrobial resistance (AMR) profiles of artisanal goat and sheep milk cheeses produced in Poland. Ten raw milk cheeses (five each from goat and sheep milk) were analyzed using a combined approach involving culture-dependent enumeration, 16S rRNA gene sequencing, and antibiotic susceptibility testing. Microbial counts revealed substantial variability among the samples, with lactic acid bacteria (LAB) dominating the microbiota. Taxonomic analysis confirmed the predominance of Lactococcus, Streptococcus, and lactobacilli, although marked intra-group heterogeneity was observed. Multivariate analyses indicated that sample-specific factors had a greater influence on microbiome composition than milk origin. Among 170 isolates, 28.7% were classified as multidrug-resistant (MDR), being most prevalent in Enterobacterales (100%) and Enterococcus spp. (73%), whereas LAB exhibited low resistance levels (16.2%). Resistance was most frequently associated with aminoglycosides and β-lactams. The resistance results were interpreted according to CLSI guidelines. These findings demonstrate that artisanal cheeses harbor complex, dynamic microbial ecosystems that may serve as reservoirs of antimicrobial resistance. The results highlight that environmental and technological factors, rather than milk source alone, are key drivers of both microbiome structure and resistance distribution, underscoring the need for targeted AMR monitoring in traditional dairy products.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Probiotics and Fermented Foods · Milk Quality and Mastitis in Dairy Cows · Animal Diversity and Health Studies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Food Technology of Plant Origin, Poznan University of Life Sciences, Wojska Polskiego 31, 60-624 Poznań, Poland

Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, Poland

Department Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznań, Poland

Institute of Forensic Genetics, Al. Mickiewicza 3/4, 85-071 Bydgoszcz, Poland

Funding

Ministry of Science and Higher Education under the project entitled “Research Network of Life Sciences Universities for the Development of the Polish Dairy Sector – Research Project”,

References (32)

Show source-provided references
1. Nayik, G.A., Jagdale, Y.D., Gaikwad, S.A., Devkatte, A.N., Dar, A.H., and Ansari, M.J. (2021). Bioactive components, health benefits and processing of goat milk: A review. Front. Nutr., 8.10.3389/fnut.2021.789117 ↗
2. Exploring the health benefits and functional properties of goat milk proteins10.1002/fsn3.3531 ↗
3. Nutritional and dietetic quality of milk and traditional cheese made from the milk of native breeds of sheep and goats10.1080/09712119.2021.2020125 ↗
4. Traditional cheeses: Rich and diverse microbiota with associated benefits10.1016/j.ijfoodmicro.2014.02.019 ↗
5. Microbial community dynamics in traditional cheeses
6. Microbial succession in artisanal cheeses
7. Fermented foods as microbial ecosystems10.1016/j.cell.2015.02.034 ↗
8. Metagenomics insights into food fermentations10.1111/1751-7915.12421 ↗
9. The rise of the Enterococcus: Beyond vancomycin resistance10.1038/nrmicro2761 ↗
10. European Food Safety Authority, and European Centre for Disease Prevention and Control (2023). The European Union Summary Report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2020/2021. EFSA J., 21, e07867.10.2903/j.efsa.2023.7867 ↗
11. Genus-Wide Assessment of Antibiotic Resistance in Lactobacillus spp10.1128/AEM.01738-18 ↗
12. Prevalence and molecular characterization of antimicrobial resistance in Escherichia coli isolated from raw milk and raw milk cheese in Egypt10.4315/0362-028X.JFP-17-277 ↗
13. Devirgiliis, C., Zinno, P., and Perozzi, G. (2013). Update on antibiotic resistance in foodborne Lactobacillus and Lactococcus species. Front. Microbiol., 4.10.3389/fmicb.2013.00301 ↗
14. Duche, R.T., Singh, A., Wandhare, A.G., Sangwan, V., Sihag, M.K., Nwagu, T.N.T., Panwar, H., and Ezeogu, L.I. (2023). Antibiotic resistance in potential probiotic lactic acid bacteria of fermented foods and human origin from Nigeria. BMC Microbiol., 23.10.1186/s12866-023-02883-0 ↗
15. Reservoirs of antimicrobial resistance genes in retail raw milk10.1186/s40168-020-00861-6 ↗
16. Bacteriocin-producing lactic acid bacteria in dairy ecosystems
17. (2004). Milk and Milk Products—Enumeration of Colony-Forming Units of Yeasts and/or Moulds—Colony-Count Technique at 25 °C (Standard No. ISO 6611:2004).
18. CLSI (2025). Performance Standards for Antimicrobial Susceptibility Testing, Clinical and Laboratory Standards Institute. [35th ed.]. CLSI Supplement M100.
19. A&A Biotechnology (2023). Genomic Mini AX Bacteria+ Spin—Instruction Manual, A&A Biotechnology.
20. Beckman Coulter (2017). AMPure XP Beads User Guide, Beckman Coulter.
21. Oxford Nanopore Technologies (2024). Ligation Sequencing Amplicons—Native Barcoding V14 (SQK-NBD114.24), Oxford Nanopore Technologies.
22. Kaczorowski, Ł., Powierska-Czarny, J., Wolko, Ł., Piotrowska-Cyplik, A., Cyplik, P., and Czarny, J. (2022). The Influence of Bacteria Causing Subclinical Mastitis on the Structure of the Cow’s Milk Microbiome. Molecules, 27.10.3390/molecules27061829 ↗
23. QIAGEN (2026). CLC Microbial Genomics Module User Manual, Version 26.0, QIAGEN Aarhus A/S.
24. Rantsiou, K., Kathariou, S., Winkler, A., Skandamis, P., and Cocolin, L. (2022). Food microbiomes and antimicrobial resistance. Front. Microbiol., 13.
25. Resistance mechanisms in Enterococcus10.4161/viru.21282 ↗
26. Emergence of epidemic multidrug-resistant Enterococcus faecium from animal and commensal strains10.1128/mBio.00534-13 ↗
27. Mobile genetic elements and AMR10.1128/CMR.00088-17 ↗
28. The resistome of pasteurized and raw milk cheeses10.1016/j.ijfoodmicro.2025.111333 ↗
29. Tracking microbial quality, safety and environmental contamination sources in artisanal goat cheesemaking factories10.1016/j.fm.2023.104301 ↗
30. Traceability, virulence and antimicrobial resistance of Yersinia enterocolitica in two industrial cheese-making plants10.1016/j.ijfoodmicro.2023.110225 ↗
31. Bayrakal, G.M., and Aydin, A. (2024). Investigation of various toxigenic genes and antibiotic and disinfectant resistance profiles of Staphylococcus aureus originating from raw milk. Foods, 13.10.20944/preprints202409.2232.v1 ↗
32. Tracking of enrofloxacin antibiotic in the making of common Middle Eastern cheeses10.1016/j.afres.2021.100004 ↗