Impact of Thermal Processing of Sardines and Sprats on Probiotic Adhesion to Intestinal Cell Models

Small pelagic fish are nutrient-dense foods, but whether domestic cooking alters their capacity to modulate probiotic adhesion is unclear. We prepared sardines and sprats using five household techniques (raw, cooked, steamed, baked, and fried) and generated in vitro digestates using the INFOGEST method. We tested two concentrations in two intestinal co-cultures—Caco-2/HT29 and mucin-producing Caco-2/HT29-MTX. Adhesion of Lacticaseibacillus rhamnosus, Lactobacillus gasseri, and Lactobacillus brevis were quantified. Digestates altered adhesion in a probiotic strain species in a process-dependent manner. Sprat digestates from fried or baked preparations produced the strongest stimulation, exceeding 150% in Caco-2/HT29; responses in HT29-MTX were directionally similar but attenuated. In contrast, cooked or steamed sardine digestates frequently inhibited adhesion, particularly at 0.5% (e.g., L. gasseri < 50%). Raw preparations yielded divergent outcomes across models. A two-way ANOVA confirmed significant effects of processing, concentration, and their interaction, with the interaction explaining up to 21% of the observed variance. Across conditions, L. rhamnosus adhered most consistently, whereas L. brevis and L. gasseri were more environmentally sensitive. These findings suggest that standard cooking practices alter the bioactivity of fish-derived digestates and, consequently, the adhesion of beneficial lactobacilli in intestinal cell models, selecting sprats and employing dry-heat methods may favor probiotic–host interactions under in vitro conditions.