Development and application of a multi-step porcine in vitro system to evaluate feedstuffs and feed additives for their efficacy in nutrient digestion, digesta characteristics, and intestinal immune responses

In vitro model provides alternatives to the use of live animals in research. In pig nutrition, there has been a tremendous increase in in vivo research over the decades. Proper utilization of in vitro models could provide a screening tool to reduce the needs of in vivo studies, research duration, cost, and the use of animals and feeds. This study aimed to develop a multi-step porcine in vitro system to simulate nutrient digestion and intestinal epithelial immune responses affected by feedstuffs and feed additives. Seven feedstuffs (corn, corn distillers dried grains with solubles corn DDGS, barley, wheat, soybean meal, soy protein concentrates, and Corynebacterium glutamicum cell mass CGCM), feed enzymes (xylanase and phytase), and supplemental amino acids (arginine, methionine, and tryptophan), were used in this in vitro evaluation for their efficacy on digestibility, digesta characteristics, and intestinal health compared with the results from previously published in vivo studies. All in vitro evaluations were triplicated. Data were analyzed using Mixed procedure of SAS9.4. Evaluations included (1) nutrient digestibility of feedstuffs; (2) the effects of feed enzymes, xylanase and phytase, on digestibility of feedstuffs and specific substrates, and (3) the effects of amino acids, arginine, tryptophan, and methionine, on anti-inflammatory, anti-oxidative, and anti-heat stress statuses showing their effects (P < 0.05) on the measured items. Differences in dry matter and crude protein digestibility among the feedstuffs as well as effects of xylanase and phytase were detected (P < 0.05), including xylo-oligosaccharide profiles and phosphorus release from phytate. Supplementation of arginine, tryptophan, and methionine modulated (P < 0.05) cellular inflammatory and oxidative stress responses. The use of this in vitro model allowed the use of 3 experimental replications providing sufficient statistical power at P < 0.05. This indicates in vitro models can have increased precision and consistency compared with in vivo animal studies.