New therapeutic molecules for farm animals are needed to address worldwide problems in the food industry, like the rise of resistance among ruminant parasites and pathogenic microbes. Since in vivo testing would involve an excessive number of animals, with consequent ethical and economic issues, the generation of sheep intestinal organoids represents a promising close-to-reality in vitro model for veterinary drug development; however, the characterization and application of such organoids remain limited. In this study, ovine intestinal organoids were generated from adult LGR5+ stem cells from the intestinal crypts of freshly slaughtered lambs, and developed in an in vitro culture system. Morphological analysis via brightfield microscopy and immunocytochemical staining revealed a pseudostratified epithelium with multiple cell types, and distinct apical–basal polarity, while RNA sequencing validated the preservation of the physiological characteristics of the original organ. The development and characterization of a robust and reproducible protocol for culturing sheep duodenum intestinal organoids in a high-throughput screening (HTS) compatible format demonstrated reliability in HTS applications, with Z’-factor tests indicating robust assay performance. Dose–response studies using pre-identified compounds showed comparable pharmacodynamic profiles between mouse and sheep organoids. These findings establish sheep intestinal organoids as an innovative tool for veterinary pharmacology and toxicology, offering a cost-effective and sustainable platform to address challenges such as drug resistance and improve livestock health.

Ovine colostrum exosomes obtained from nutritionally programmed dairy ewes (F0) may present modifications in microRNAs, thus having consequences for the intestinal barrier function and immunity parameters of lambs (F1). To test this hypothesis, colostrum exosomes from two ewe groups [F0-MET (nutritionally programmed ewes being fed methionine during early life) and F0-CTRL (ewes not supplemented with methionine during early life)] were sequenced to compare differences in the miRNAome. In addition, these exosomes were added to an in vitro co-culture in a Transwell chamber system consisting of ovine duodenum intestinal organoids and macrophages to assess the expression of genes encoding tight junction proteins in organoids and immunity parameters in macrophages. Finally, the concentrations of cytokines (e.g., IL-12 and IL-6) were assessed by ELISA kits in the supernatants of the chamber containing macrophages. According to the miRNAome, the expression of two miRNAs (e.g., oar_miR_376c_3p and oar_miR_432) was reduced in the colostrum exosomes obtained from dairy ewes nutritionally programmed with dietary supplementation of methionine during early life (F0-MET ewes). These changes did not seem to modify the expression of intestinal barrier and immune response marker genes when these exosomes were added to a co-culture of ovine intestinal organoids and macrophages. However, the levels of IL-12 produced by macrophages were reduced (p < 0.05), which suggests the inhibition of inflammatory pathways. Further studies using ovine colostrum exosomes obtained from nutritionally programmed ewes will help to clarify their potential to improve the health of suckling lambs.

Adequate management of replacement ewe lambs (F0) in dairy sheep farms during postnatal life may modify the germline cells, thus promoting transmission of intergenerational effects to the offspring (F1). To test this hypothesis, 18 newborn male lambs (F1), either born from methionine-supplemented ewe lambs (F0 ewe lambs being fed ad libitum with a milk replacer supplemented with 1 g methionine/kg DM) or not supplemented (F0 ewe lambs being fed ad libitum with the same milk replacer with no methionine added), were included in the present study. All the male F1 lambs were managed exactly in the same way along the whole lifespan in order to bring out the differences caused by methionine supplementation of F0 dams. Our data show that the methionine supplementation of dams (F0) during the suckling period did not promote significant (p > 0.05) changes on feed intake, growth rate, or feed efficiency of F1 male lambs during the fattening period. Moreover, the meat chemical composition (proximal, fatty acid profile, and volatile compounds) was similar for both groups (p > 0.05), but the meat of F1-MET lambs presented higher redness and hardness (p < 0.05) when compared to F1-CTRL lambs. The biochemical profile also highlighted significant (p < 0.05) differences in the serum creatinine and calcium content that may be at least partially related to the meat quality traits observed. Overall, all these results suggest that methionine supplementation of lambs (F0) during early postnatal life causes permanent changes in the offspring. This has positive effects, such as achieving a more attractive color of lamb meat (F1) for consumers, and negative effects, such as reduced meat tenderness.