Nicole Maculet

Uterine physiology may differ among cattle managed under different farming systems. This pilot study evaluated uterine NAMPT/visfatin and SPP1/osteopontin in pregnant and luteal-phase non-pregnant cattle from one intensive beef farm (INT; n = 7; nulliparous heifers aged 14–18 months, crossbred with a predominant Limousin genetic background) and one extensive farm (EXT; n = 6; Lidia cows aged 28–34 months). Formalin-fixed paraffin-embedded uterine tissues were analyzed by RT-qPCR and immunohistochemistry. Visfatin immunoreactivity was mainly localized in the luminal epithelium and uterine glands. NAMPT expression was higher in the INT cohort than in the EXT cohort (0.6331 ± 0.0855 vs. 0.3632 ± 0.0395; p = 0.0200), whereas SPP1 expression did not differ between cohorts (0.2056 ± 0.0661 vs. 0.1806 ± 0.0562; p = 0.7786). Visfatin immunostaining scores were significantly higher in INT than EXT cows in the secretory adenomeres (median [IQR]: 2.37 [2.15–2.52] vs. 1.48 [1.38–1.77]; p = 0.0012), whereas no significant difference was detected in the combined luminal epithelium/glandular duct compartment (p = 0.4639). Because age, breed, and farm of origin were confounded with management system, these findings should be interpreted as exploratory between-cohort associations.

Grape pomace (GP) is being explored as a sustainable feed ingredient for ruminants due to its fiber and polyphenol content; however, its systemic effects under farming conditions remain unclear. This study tested whether 5% dried grape pomace (GP5) modulates serum circulating miRNAs (cmiRNAs) in dairy sheep. Twelve Sarda ewes were enrolled in a 30-day trial and assigned to a control (CTRL) or GP5 diet. Small RNA sequencing was initially performed in GP5 animals (paired T30 vs. T0 comparison); after quality filtering, five paired animals were included in the sequencing analyses. The differentially expressed (DE) cmiRNAs were validated by RT-qPCR in GP5 and CTRL groups. RT-qPCR confirmed the NGS-derived miRNA signature, identifying miR-143, miR-382-5p, miR-409-3p, and miR-194 as significantly downregulated (p < 0.05) in GP5 animals. Ingenuity Pathway Analysis (IPA) showed that DE miRNAs target shared hubs, with PPAR signaling among the most enriched pathways (BH-adjusted p = 1.35 × 10⁻⁹), predicted to be inhibited (Z = −2.646). Cholecystokinin/gastrin-mediated and S100 signaling were also enriched (BH-adjusted p = 7.08 × 10⁻¹¹ and 3.21 × 10⁻⁸) and predicted to be activated (Z = 2.828 and 3.162). Overall, GP5 supplementation was associated with a selective cmiRNA signature consistent with targeted post-transcriptional regulation.