Apple Pomace and Performance, Intestinal Morphology and Microbiota of Weaned Piglets-A Weaning Strategy for Gut Health?

Apple pomace (AP) is known to be rich in biomolecules beneficial for health and it may advantageously be used to overcome the critical step of piglets' weaning. The study aimed to determine the effect of two levels of incorporation of AP on the performance, intestinal morphology, and microbiota of weaned piglets and investigate this feed ingredient as a weaning strategy. An experiment was performed with 42 piglets from weaning (28 days old) over a five-week period, including three iso-energetic and iso-nitrogenous diets (0%, 2%, and 4% dried AP diets) with seven pen-repetitions per diet (two pigs per pen). AP diets were beneficial for the average daily gain calculated on week 3 (p = 0.038) and some parameters of the intestinal architecture on the 35 post-weaning day. The 4% AP diet was beneficial for the feed conversion ratio (p = 0.002) and the energetic feed efficiency (p = 0.004) on the 35 post-weaning day. AP tended to influence the consistency of feces (softer to liquid, p = 0.096) and increased the counts of excreted pathogens (p = 0.072). Four percent AP influenced the richness of the microbiota and the bacteria profile as observed for the phylum Bacteroidetes or the class Clostridia. The 4% AP diet appeared as an interesting weaning strategy that should be evaluated in a large cohort.

Baby-SPIME: A dynamic in vitro piglet model mimicking gut microbiota during the weaning process.

The study aimed to adapt the SHIME® model, developed to simulate human digestion and fermentation, to a baby-SPIME (baby Simulator of Pig Intestinal Microbial Ecosystem). What constitutes a unique feature of this model is its twofold objective of introducing an ileal microbial community and mimicking a dietary weaning transition. This model should then be ideally suited to test the dietary weaning strategies of piglets in vitro. Regarding the microbiota, the main phyla making up the model were Firmicutes, Bacteroidetes and Proteobacteria although Bacteroidetes decreased after inoculation (p = 0.043 in ileum, p = 0.021 in colon) and Delta-Proteobacteria were favoured (p = 0.083 in ileum, p = 0.043 in colon) compared to Gamma-Proteobacteria. The designed model led to a low representation of Bacilli - especially Lactobacillus sp. in the ileum and a weak representation of Bacteroidia in the proximal colon. However, Mitsuokella and Prevotella were part of the major genera of the model along with Bifidobacterium, Fusobacterium, Megasphaera and Bacteroides. As a result of weaning, two major changes - normally occurring in vivo - were detected in the system: firstly, Firmicutes diminished when Bacteroidetes increased particularly in the proximal colon; secondly, Bacteroides decreased and Prevotella increased (mean value for four runs). In terms of metabolite production, while a ratio acetate: propionate: butyrate of 60:26:14 was obtained in post-weaned colon, the expected inversion of the ratio propionate: butyrate in the post-weaned ileum was unfortunately not observed. To conclude, the so-called baby-SPIME model meets expectations regarding the resident microbiota of the proximal colon bioreactor and the metabolites produced thereof. In terms of the evolution of major groups of bacteria, the in vitro weaning process appeared to be successful. However, higher concentration of butyric acid would have been expected in ileum part of newly weaned piglets, as observed in vivo. The microbiota in the ileum bioreactor seemed in fact to act like a pre-colon. This suggests that microbial profile in ileum bioreactor had to be improved.

Development of an analytical method to detect short-chain fatty acids by SPME-GC-MS in samples coming from an in vitro gastrointestinal model.

Short chain fatty acids (SCFA) are end-products of intestinal bacterial fermentation. The concentrations of fermentation metabolites are closely related to the microbial activity that occurs in various digestive compartments. The fermentation products may vary qualitatively and quantitatively, especially within the colon. The Simulator of the Human Intestinal Microbial Ecosystem (SHIME), an in vitro dynamic and multicompartment model of the human intestinal tract, can be adapted to mimic the piglet gastrointestinal tract. In this context, a quantitative method, based on solid phase microextraction gas chromatography coupled to mass spectrometry (SPME-GC-MS), was developed for the determination of seven short chain fatty acids, i.e. acetic, propionic, butyric, isobutyric, valeric, isovaleric and hexanoic acids, in samples coming from this experimental in vitro gastrointestinal model. The advantage of the SPME-GC-MS technique is that the seven compounds could be determined in a single run, after a simple and rapid sample treatment, without any other extraction than the automatic SPME. The developed method was validated in accordance to the European and US FDA guidelines and showed good specificity/selectivity. In addition, limits of detection and quantification ranged from 8 to 72 mg L-1 and from 16 to 144 mg L-1, respectively. Two internal quality control samples spiked at different concentrations were analyzed to assess the trueness of the developed method, which ranged between 97.7 and 122.4% of the expected value, for the seven compounds analyzed. The method was successfully applied to twenty samples coming from a gastrointestinal model, with different inocula. The developed method might be used as a general method for measuring SCFA in biological samples.