Numerous bioactive plant additives have shown various positive effects in pigs and chickens. The demand for feed additives of natural origin has increased rapidly in recent years to support the health of farm animals and thus minimize the need for antibiotics and other drugs. Although only in vivo experiments can fully represent their effect on the organism, the establishment of reliable in vitro methods is becoming increasingly important in the goal of reducing the use of animals in experiments. The use of cell models requires strict control of the experimental conditions so that reliability and reproducibility can be achieved. In particular, the intestinal porcine epithelial cell line IPEC-J2 represents a promising model for the development of new additives. It offers the possibility to investigate antioxidative, antimicrobial, anti- or pro-proliferative and antiviral effects. However, the use of IPEC-J2 is limited due to its purely epithelial origin and some differences in its morphology and functionality compared to the in vivo situation. With regard to chickens, the development of a reliable intestinal epithelial cell model has attracted the attention of researchers in recent years. Although a promising model was presented lately, further studies are needed to enable the standardized use of a chicken cell line for testing phytogenic feed additives. Finally, co-cultivation of the currently available cell lines with other cell lines and the development of organoids will open up further application possibilities. Special emphasis was given to the IPEC-J2 cell model. Therefore, all publications that investigated plant derived compounds in this cell line were considered. The section on chicken cell lines is based on publications describing the development of chicken intestinal epithelial cell models.

The supplementation of feed with phytases enables broilers to utilize more efficiently phosphorus (P) from phytic acid (IP₆), the main storage form of P in plants. The current study evaluated the addition of 500, 1000, and 3000 FTU of phytase per kg to a phytate-containing diet with low P level (LP) fed to broilers from 1 to 21 days of age and compared it to a hypoallergenic phytate-free diet (HPF). There was a linear improvement in performance parameters with increasing levels of phytase in the LP diet (p < 0.001). Apparent ileal digestibility of crude protein, P, and some amino acids, increased with phytase. Crude ash, P, and the calcium content of tibia bones linearly increased with increasing levels of phytase (p < 0.001). Crypt depth (related to body weight) in the jejunum epithelium linearly decreased with phytase addition (p < 0.001). Cecal crypt depth decreased with phytase supplementation (p = 0.002). Cecum tissue showed lower counts of CD3-positive intraepithelial lymphocytes in broilers receiving the phytase in comparison to LP (p < 0.001), achieving similar counts to HPF-fed broilers. Although results from the current study seem to point out some mechanisms related to the immune response and mucosal morphology contributing to those overall beneficial effects, no clear differences between different phytase doses could be demonstrated in these specific parameters.

Rye and rapeseed meal (RSM) are alternatives to wheat and soybean meal (SBM) in pig nutrition. The inclusion of rye and RSM instead of wheat and SBM may increase dietary fibre content in the feed, which may affect digestive physiology and intestinal function. However, research on the combined feeding of rye and RSM in piglets is still scarce. The aim of this study was to investigate the effects of rye and RSM containing feed on growth performance, protein and amino acid digestibility, physical characteristics of the digesta and the intestinal morphology in weaner piglets. A total of 88 piglets aged 28 d were randomly allotted to four different feeding groups at weaning: wheat/SBM, wheat/RSM, rye/SBM, rye/RSM. Cereals were included at 48%, SBM at 25% and RSM at 30%. Body weight gain and feed intake were recorded weekly. After 33 d, piglets were euthanised for assessment of digesta and intestinal morphology. Rye did not affect performance, whilst RSM resulted in reduced weight gain (p = 0.024) and feed intake (p = 0.037). Apparent ileal digestibility of crude protein and amino acids was decreased by rye and RSM (p < 0.05). Rye caused an acidification (p = 0.045) and increased viscosity (p = 0.013) of small intestinal digesta, a lower dry matter content of jejunal and colonic digesta (p < 0.05) and an increased relative weight of the colon tissue (p < 0.001). In conclusion, performance was not reduced when rye was used as an alternative to wheat in diets for weaner piglets. The inclusion of RSM instead of SBM to piglets’ diet lowered animal performance under our conditions. Further investigations are warranted to characterise the quality of rapeseed meal and the optimum inclusion level in the diet for young pigs.

Clostridioides difficile toxins are one of the main causative agents for the clinical symptoms observed during C. difficile infection in piglets. Porcine milk has been shown to strengthen the epithelial barrier function in the piglet’s intestine and may have the potential to neutralise clostridial toxins. We hypothesised that porcine colostrum exerts protective effects against those toxins in the IPEC-J2 cells and in the colon epithelium of healthy piglets. The IPEC-J2 cells were treated with either the toxins or porcine colostrum or their combination. Analyses included measurement of trans-epithelial electrical resistance (TEER), cell viability using propidium iodide by flow cytometry, gene expression of tight junction (TJ) proteins and immune markers, immunofluorescence (IF) histology of the cytoskeleton and a TJ protein assessment. Colon tissue explants from one- and two-week-old suckling piglets and from five-week-old weaned piglets were treated with C. difficile toxins in Ussing chamber assays to assess the permeability to macromolecules (FITC-dextran, HRP), followed by analysis of gene expression of TJ proteins and immune markers. Toxins decreased viability and integrity of IPEC-J2 cells in a time-dependent manner. Porcine colostrum exerted a protective effect against toxins as indicated by TEER and IF in IPEC-J2 cells. Toxins tended to increase paracellular permeability to macromolecules in colon tissues of two-week-old piglets and downregulated gene expression of occludin in colon tissues of five-week-old piglets (p = 0.05). Porcine milk including colostrum, besides other maternal factors, may be one of the important determinants of early immune programming towards protection from C. difficile infections in the offspring.

Epizootic Rabbit Enteropathy (ERE) is a disease of unknown aetiology that mainly affects post-weaning animals. Caecotrophs from animals in a farm affected by ERE were analysed to identify changes in the microbiological profile of growing rabbits. Does and kits at weaning (28 d) and the same rabbits ten days later (38 d) were used for a comparison using Roche 454 pyrosequencing of hypervariable V3-V5 regions of the 16S rRNA genes. The caecal bacterial community was dominated by the Firmicutes phylum (about 80%), followed by Bacteroidetes (15%), although relative abundances changed according to animal age (among does and kits at 28 and 38 d) and health status (affected or not by ERE). Two dominant families were classified within the Firmicutes phylum: Ruminococcaceae and Lachnospiraceae (50 and 20% of the sequences, respectively). In kits affected by ERE, relative abundance of Ruminococcus and Bacteroides genera decreased and increased, respectively, compared to healthy kits at the same age (28 and 38 d). The principal coordinate analysis plot revealed that kits at 28 d of age cluster together and apart from the does and the healthy 38-d rabbit groups. When only growing rabbits are considered, kits that showed symptoms of ERE clustered separately. Results suggest a different caecal bacterial community of rabbits affected by ERE. These findings highlight the need to identify different stages of the disease.