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Peer-reviewedCC BY 4.0Veterinary Research

A single-spheroid bovine mammary co-culture model for longitudinal monitoring and preclinical safety assessment of antimicrobial alternatives against mastitis-causing pathogens

  • Julie Bellemans1,2
  • Yves Briers2
  • Laurence Finot3
  • Evelyne Meyer1
  1. 1Laboratory of Biochemistry, Department of Veterinary and Biosciences, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium
  2. 2Laboratory of Applied Biotechnology, Department of Biotechnology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
  3. 3PEGASE, INRAE, Institut Agro Rennes-Angers, 35590 Saint-Gilles, France
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Abstract

Bovine mastitis remains a major driver of antimicrobial use in dairy production, highlighting the need for physiologically relevant in vitro models for preclinical safety assessment of novel intramammary therapeutics. Here, a previously established bovine epithelial cell line co-culture mammosphere model was adapted to a single-spheroid 96-well ultra-low attachment format, improving reproducibility and longitudinal monitoring. Mammospheres maintained epithelial organization and were further characterized for secretory function through assessment of casein expression. The suitability of the model for safety assessment was subsequently evaluated using complementary fluorescence microscopy, kinetic live-cell imaging, and flow cytometry following Triton X-100-induced cytotoxicity. Using this workflow, the safety of bacteriophage-derived recombinant lysin LysMK34 and antibiotic kanamycin, alone and in combination, was evaluated, with no cytotoxic effects observed, supporting the utility of the model for future preclinical intramammary safety assessment.

Supplementary information: The online version contains supplementary material available at https://doi.org/10.1186/s13567-026-01849-0.

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Cite

BibTeXRIS
BibTeX
@article{bellemans2026singlespheroid,
  title = {{A single-spheroid bovine mammary co-culture model for longitudinal monitoring and preclinical safety assessment of antimicrobial alternatives against mastitis-causing pathogens}},
  author = {Julie Bellemans and Yves Briers and Laurence Finot and Evelyne Meyer},
  journal = {Veterinary Research},
  year = {2026},
  month = nov,
  doi = {10.1186/s13567-026-01849-0},
  url = {https://doi.org/10.1186/s13567-026-01849-0},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - A single-spheroid bovine mammary co-culture model for longitudinal monitoring and preclinical safety assessment of antimicrobial alternatives against mastitis-causing pathogens
AU  - Julie Bellemans
AU  - Yves Briers
AU  - Laurence Finot
AU  - Evelyne Meyer
PY  - 2026
DA  - 2026/11/04
JO  - Veterinary Research
DO  - 10.1186/s13567-026-01849-0
UR  - https://doi.org/10.1186/s13567-026-01849-0
AB  - Bovine mastitis remains a major driver of antimicrobial use in dairy production, highlighting the need for physiologically relevant in vitro models for preclinical safety assessment of novel intramammary therapeutics. Here, a previously established bovine epithelial cell line co-culture mammosphere model was adapted to a single-spheroid 96-well ultra-low attachment format, improving reproducibility and longitudinal monitoring. Mammospheres maintained epithelial organization and were further characterized for secretory function through assessment of casein expression. The suitability of the model for safety assessment was subsequently evaluated using complementary fluorescence microscopy, kinetic live-cell imaging, and flow cytometry following Triton X-100-induced cytotoxicity. Using this workflow, the safety of bacteriophage-derived recombinant lysin LysMK34 and antibiotic kanamycin, alone and in combination, was evaluated, with no cytotoxic effects observed, supporting the utility of the model for future preclinical intramammary safety assessment.  Supplementary information: The online version contains supplementary material available at https://doi.org/10.1186/s13567-026-01849-0.
SN  - 0928-4249
SN  - 1297-9716
ER  -