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Probiotics have been considered as a promising strategy to prevent various diseases in both humans and animals. This approach has gained interest in recent years as a potential means to control bovine mastitis. In a previous study, we found that several  L .  casei  strains, including BL23, were able to inhibit the internalization of  S .  aureus , a major etiologic agent of mastitis, into bovine mammary epithelial cells (bMEC). This antagonism required a direct contact between  L .  casei  and bMEC or  S .  aureus , suggesting the inhibition relied on interactions between  L .  casei  cell surface components and bMEC. In this study, we have investigated the impact of some candidates which likely influence bacteria host cell interactions. We have shown that  L .  casei  BL23  fbpA  retained its inhibitory potential, indicating that  L .  casei  BL23 antagonism did not rely (solely) on competition between  S .  aureus  and  L .  casei  fibronectin-binding proteins for adhesion to bMEC. We have then investigated the impact of four sortase mutants,  srtA1 ,  srtA2 ,  srtC1  and  srtC2 , and a double mutant ( srtA1-srtA2 ) on  L .  casei  BL23 inhibitory potential. Sortases are responsible for the anchoring on the bacterial cell wall of LPXTG-proteins, which reportedly play an important role in bacteria-host cell interaction. All the  srt  mutants tested presented a reduced inhibition capacity, the most pronounced effect being observed with the  srtA2  mutant. A lower internalization capacity of  L .  casei srtA2  into bMEC was also observed. This was associated with several changes at the surface of  L .  casei  BL23  srtA2  compared to the wild type (wt) strain, including altered abundance of some LPXTG- and moonlighting proteins, and modifications of cell wall structure. These results strongly support the role of sortase A2 in  L .  casei  BL23 inhibition against  S .  aureus  internalization. Deciphering the contribution of the cell surface components altered in  srtA2  strain in the inhibition will require further investigation.

Contribution of sortase SrtA2 to Lactobacillus casei BL23 inhibition of Staphylococcus aureus internalization into bovine mammary epithelial cells