Search

IntroductionThe aim of this study was to evaluate the effect of two MS formulas, DanMilk™ (AB Neo, Denmark) (MS1) and Neopigg® RescueMilk (Provimi, Netherlands) (MS2) administered manually and to compare two ways of administration (manual vs automatic) of MS1 on growth performance, health, fecal microbial profile, behavior, and skin lesions of piglets during suckling and post-weaning.MethodsForty litters (528 piglets) were divided into 4 groups: 1) Control group receiving no MS (CON); 2) MS1 administered automatically (A-MS1); 3) MS1 administered manually (Ma-MS1) 4) MS2 administered manually (Ma-MS2). All groups had access to sow milk and creep feed. On day 5 after birth (d0), litters were equalized (13.2 piglets/litter ± 0.8 SD), thereafter no cross-fostering was allowed. Piglets were weighed at day 5 after birth (d0), at the end of milk supplementation (d14), at weaning (d21 of the trial, 26 days of age) and ten days post-weaning (d31). Piglet welfare was assessed using behavioral and lesion measures at d4 and d10. Feces were collected at d14 and d21.Results and discussionDuring the suckling period, A-MS1 had lowest mortality (p < 0.05), while Ma-MS1 had lower mortality compared with CON and Ma-MS2 (p < 0.05). Negative social behavior at d4, was more frequent in MS groups (A-MS1, Ma-MS1, Ma-MS2) compared to CON group (p = 0.03). Growth performance and lesion prevalence were not affected by MS provision. During lactation, Ma-MS2 group had a higher percentage of piglets not eating during suckling at d18 compared with Ma-MS1 (p = 0.03). MS1 increased microbial diversity compared with CON at d14 (Chao1, p = 0.02; Shannon, p = 0.03) and compared with CON (Shannon, p < 0.05; InvSimpson, p = 0.01) and Ma-MS2 (Chao1, p < 0.05; Shannon, p = 0.05, InvSimpson p = 0.01) at d21. Groups that received MS1 were characterized by genera producing short-chain fatty acids (SCFAs), i.e., Lachnospiraceae (A-MS1) and Oscillospiraceae (Ma-MS1). MS composition and availability can contribute to reduce piglet's mortality during the suckling phase and can also affect intestinal microbiota by favoring the presence of SCFAs producing bacteria.

Additional file 5: Figure S5. β-diversity of the mucosa-associated microbiota, milk, skin, vagina and feces of sows, and environmental microbiota. PCoA of phylogenetic community composition based on weighted UniFrac (A), and taxonomic community composition based on Bray–Curtis (B).

Table S3. Effect of the dietary supplementations of Bacillus amyloliquefaciens or Bacillus subtilis, on the jejunum histology and morphology of weaned pigs 14 days post-infection with ETEC F4. (DOCX 39 kb)

Figure S1. Non-Metric Multidimensional Scaling (NMDS) plot on Bray-Curtis distance matrix in jejunum and cecum of ETEC F4-infected pigs fed supplementations of Bacillus amyloliquefaciens DSM25840, Bacillus subtilis DSM25841 and antibiotic at d 21. (DOCX 521 kb)

Table S2. List of number of Observed OTUs, Shannon index and number of reads in jujunum and cecum samples. (DOCX 34 kb)

Additional file 7: Figure S7. Distance comparison of microbial communities between true compared with random sow-piglet pairs for fecal, vaginal and environmental microbiota (Values are Means ± SE; significance between intestinal segments was determined by pairwise Kruskal-Wallis test; * P < 0.05; ** P < 0.01; *** P < 0.001).

Additional file 6: Figure S6. Distance comparison of microbial communities between mucosa-associated intestinal microbiota with that of maternal milk and feces. Values are Means ± SE; significance between intestinal segments was determined by pairwise Kruskal-Wallis test; * P < 0.05; ** P < 0.01; *** P < 0.001.

The main bins accounting for the spectral differentiation and their relative chemical shift. (DOCX 21 kb)

The main bins accounting for the spectral differentiation and their relative chemical shift. (DOCX 21 kb)

We tested the effect of Trp addition to a standard weaning diet and oral challenge with enterotoxigenic Escherichia coli K88 (ETEC) on growth and health of piglets susceptible or nonsusceptible to the intestinal adhesion of ETEC. Sixty-four pigs weaned at 21 d of age were divided into 3 groups based on their ancestry and BW: a control group of 8 pigs fed a basal diet (B), the first challenged group of 28 pigs fed B diet (BCh), and the second challenged group of 28 pigs fed a diet with Trp (TrpCh). The Trp diet was produced by the addition of 1 g of l-Trp/kg to the basal diet. On d 5, pigs were orally challenged with 1.5 mL suspension containing 10(10) cfu ETEC/mL or placebo, and killed on d 9 or 23. Based on in vitro villus adhesion assay, the pigs (except the B group) were classified as susceptible (s(+)) or nonsusceptible (s(-)) to the intestinal ETEC adhesion. Thus, after the challenge, treatments were B, BChs(-), BChs(+), TrpChs(-), and TrpChs(+). Pigs susceptible to ETEC were 50.0% in the BChs(+) group (3 pigs lost included) and 46.4% in the TrpChs (+) group (1 pig lost included). During the first 4 d after challenge, the challenge reduced ADG (P0.05), and this reduction was greater in susceptible pigs (P0.05) than nonsusceptible ones. Tryptophan increased ADG and feed intake in susceptible pigs (P0.05) from challenge to d 4, but not thereafter. Tryptophan supplementation did not improve the fecal consistency and did not reduce the number of pigs positive for ETEC in feces on d 4 after the challenge. The K88-specific immunoglobulin A activity in blood serum tended to be greater in challenged pigs (P = 0.102) and was not affected by the addition of Trp. Villous height was affected by the addition of Trp and challenge in different ways, depending on the site of small intestine. The need to consider the phenotype for the adhesion of the ETEC in studies with different supply of Trp was clearly evident. When compared with practical weaning standard diets, Trp supplementation allowed susceptible pigs to partially compensate for the effects of ETEC challenge by increasing feed intake and maintaining an adequate BW growth. This is of practical importance for the formulation of diets for pigs selected for lean growth because of the presence of an association between this trait and the susceptibility to the intestinal adhesion of ETEC.

Sodium butyrate (SB) is used as an acidifier in animal feed. We hypothesized that supplemental SB impacts gastric morphology and function, depending on the period of SB provision. The effect of SB on the oxyntic and pyloric mucosa was studied in 4 groups of 8 pigs, each supplemented with SB either during the suckling period (d 4 to d 28 of age), after weaning (d 29 to d 39-40 of age) or both or never. The number of parietal cells immunostained for H+/K+-ATPase, gastric endocrine cells immunostained for chromogranin A and somatostatin (SST) in the oxyntic mucosa and gastrin-secreting cells in the pyloric mucosa, was assessed. Gastric muscularis and mucosa thickness were measured. Expressions of the H+/K+-ATPase and somatostatin type 2 receptor (SSTR2) genes in the oxyntic mucosa, and of the gastrin gene in the pyloric mucosa were evaluated by real-time RT-PCR. SB increased the number of parietal cells per gland, regardless of the period of administration (P < 0.05). SB addition after, but not before, weaning increased the number of enteroendocrine and SST-positive cells (P < 0.01) and tended to increase gastrin mRNA (P = 0.09). An interaction between the two periods of SB treatment was seen for the expression of H/K-ATPase and SSTR2 genes (P < 0.05). Butyrate intake after weaning increased gastric mucosa thickness (P < 0.05), but not muscularis. Sodium butyrate used orally at a low dose impacted gastric morphology and function presumably in relationship with its action on mucosal maturation and differentiation.

Additional file 1: Figure S1. Experimental design and workflow of sample collection. Each tick denotes a time-point of sample collection.

Additional file 1: Figure S1. Experimental design and workflow of sample collection. Each tick denotes a time-point of sample collection.

Additional file 3: Figure S3. (A) Negligible influence of breed on the mucosa-associated microbiota at birth. Average weighted UniFrac, unweighted UniFrac and Bray-Curtis distance between individuals at birth within and between Rongchang and Landrace piglets. (B) Shift of α diversity of mucosa-associated microbiota with age across four intestinal segments based on Chao1 index (Values are Means ± SE; significance between groups was determined by Kruskal-Wallis test).

Additional file 2: Table S1. Effect of the season of farrowing on the relative abundance of the oligosaccharides that were detected in at least 50% of the samples. Table S2. Effect of the interaction between the breed and the season of farrowing on the relative abundance of the oligosaccharides that were detected in at least 50% of the samples. Figure S1. Principal Component Analysis plot of oligosaccharides values for the three breeds (DU = Duroc; LA = Landrace; LW = Large White): Plot A) Principal Component 1 (Prin1) vs. 2 (Prin2); Plot B) Principal Component 1 (Prin1) vs. 3 (Prin3).

Additional file 3: Figure S3. (A) Negligible influence of breed on the mucosa-associated microbiota at birth. Average weighted UniFrac, unweighted UniFrac and Bray-Curtis distance between individuals at birth within and between Rongchang and Landrace piglets. (B) Shift of α diversity of mucosa-associated microbiota with age across four intestinal segments based on Chao1 index (Values are Means ± SE; significance between groups was determined by Kruskal-Wallis test).

Additional file 2: Table S1. Effect of the season of farrowing on the relative abundance of the oligosaccharides that were detected in at least 50% of the samples. Table S2. Effect of the interaction between the breed and the season of farrowing on the relative abundance of the oligosaccharides that were detected in at least 50% of the samples. Figure S1. Principal Component Analysis plot of oligosaccharides values for the three breeds (DU = Duroc; LA = Landrace; LW = Large White): Plot A) Principal Component 1 (Prin1) vs. 2 (Prin2); Plot B) Principal Component 1 (Prin1) vs. 3 (Prin3).

Additional file 2: Figure S2. Rarefaction curves based on the Chao1 and Shannon index at increasing sequencing depth of intestinal mucosal, maternal and environmental samples.

The aim of this study was to test the effect of mixing doses of glutamate (Glu) and glutamine (Gln) on the growth, health and gut health of post-weaning piglets. One hundred twenty weaned piglets (24 ± 2 days of age) were assigned to 6 dietary groups: (1) standard diet (CO); (2) CO plus Glu (6 kg/Ton): 100Glu; (3) CO plus 75Glu + 25Gln; (4) CO plus 50Glu + 50Gln; (5) CO plus 25Glu + 75Gln and (6) CO plus 100Gln. At days 8 and 21, blood was collected for haematological and reactive oxygen metabolite analysis, intestinal mucosa for morphological and gene expression analysis, and caecal content for microbial analysis. Data were fitted using a Generalised Linear Model (GLM). Piglet growth increased linearly with an increase in Gln from d7 to d14. The Glu:Gln ratio had a quadratic effect on faecal consistency and days of diarrhoea, neutrophil% and lymphocyte%, and a positive linear effect on monocyte% in the blood at d8. The amino acids (AAs) reduced the intraepithelial lymphocytes in the jejunum, and 100Gln improved intestinal barrier integrity at d8. The caecal microbiota did not differ. Overall, this study suggested a favourable effect of mixing Glu and Gln (25 + 75–50 + 50) as a dietary supplementation in post-weaning piglets to benefit the immune and barrier function of the gut, resulting in an increase in faecal consistency and improvement of growth during the first 2 weeks post-weaning.

Several studies highlighted a correlation between folic acid deficiency and high plasma homocysteine concentration, considered a risk factor for multifactorial diseases. Natural folates represent an emerging alternative strategy to supplementation with synthetic folic acid, whose effects are controversial. The present work was, therefore, performed in hyperhomocysteinemic mice to study the impact of supplementation with dairy matrices containing natural folates on plasma homocysteine levels and faecal microbiota composition. Forty mice were divided into six groups, two of which fed control or folic acid deficient (FD) diets for 10 weeks. The remaining four groups were fed FD diet for the first 5 weeks and then shifted to a standard control diet containing synthetic folic acid (R) or a FD diet supplemented with folate-enriched fermented milk (FFM) produced by selected lactic acid bacteria, fermented milk (FM), or milk (M), for additional 5 weeks. Supplementation with dairy matrices restored homocysteine levels in FD mice, although impacting differently on hepatic S-adenosyl-methionine levels. In particular, FFM restored both homocysteine and S-adenosyl-methionine levels to the control conditions, in comparison with FM and M. Next generation sequencing analysis revealed that faecal microbiota of mice supplemented with FFM, FM and M were characterised by a higher richness of bacterial species in comparison with C, FD and R groups. Analysis of beta diversity highlighted that the three dairy matrices determined specific, significant variations of faecal microbiota composition, while hyperhomocysteinemia was not associated with significant changes. Overall, the results represent a promising starting point for the applicability of food matrices enriched in natural folates to manage hyperhomocysteinemia.

Background Probiosis is considered a potential strategy to reduce antibiotics use and prevent post-weaning diarrhea (PWD). This study investigated the effect of Bacillus amyloliquefaciens DSM25840 or Bacillus subtilis DSM25841 supplementation on growth, health, immunity, intestinal functionality and microbial profile of post-weaning pigs after enterotoxigenic E. coli (ETEC) F4 challenge. Methods Sixty-four post-weaning piglets (7748 g ± 643 g) were randomly allocated to four groups: control basal diet (CO); CO + 1.28 × 106 CFU/g of B. amyloliquefaciens (BAA); CO + 1.28 × 106 CFU/g feed of B. subtilis (BAS); CO + 1 g colistin/kg of feed (AB). At day (d) 7, animals were challenged with 105 CFU/mL of ETEC F4ac O149 and then followed for fecal score and performance until d 21. Blood was collected at d 6, d 12 and d 21 for immunoglobulins, at d 8 for acute phase proteins, at d 8 and d 21 for metabolomics analysis. Jejunum was sampled for morphometry, quantification of apoptosis, cell proliferation, neutral and acid mucine and IgA secretory cells, and microarray analysis at d 21. Jejunum and cecum contents were collected for microbiota at d 21. Results AB and BAS reduced the fecal score impairment compared to CO (P Bacillus groups and CO. AB improved BW at d 7, d 14 and d 21, ADWG ADFI and G:F from d 0 to d 7 (P P P = 0.06). Conclusion Bacillus subtilis DSM25841 treatment may reduce ETEC F4ac infection in weaned piglets, decreasing diarrhea and influencing mucosal transcriptomic profile.

Background The early-life microbiota exerts a profound and lifelong impact on host health. Longitudinal studies in humans have been informative but are mostly based on the analysis of fecal samples and cannot shed direct light on the early development of mucosa-associated intestinal microbiota and its impact on GI function. Using piglets as a model for human infants, we assess here the succession of mucosa-associated microbiota across the intestinal tract in the first 35 days after birth. Results Although sharing a similar composition and predicted functional profile at birth, the mucosa-associated microbiome in the small intestine (jejunum and ileum) remained relatively stable, while that of the large intestine (cecum and colon) quickly expanded and diversified by day 35. Among detected microbial sources (milk, vagina, areolar skin, and feces of sows, farrowing crate, and incubator), maternal milk microbes were primarily responsible for the colonization of the small intestine, contributing approximately 90% bacteria throughout the first 35 days of the neonatal life. Although maternal milk microbes contributed greater than 90% bacteria to the large intestinal microbiota of neonates upon birth, their presence gradually diminished, and they were replaced by maternal fecal microbes by day 35. We found strong correlations between the relative abundance of specific mucosa-associated microbes, particularly those vertically transmitted from the mother, and the expression levels of multiple intestinal immune and barrier function genes in different segments of the intestinal tract. Conclusion We revealed spatially specific trajectories of microbial colonization of the intestinal mucosa in the small and large intestines, which can be primarily attributed to the colonization by vertically transmitted maternal milk and intestinal microbes. Additionally, these maternal microbes may be involved in the establishment of intestinal immune and barrier functions in neonates. Our findings strengthen the notion that studying fecal samples alone is insufficient to fully understand the co-development of the intestinal microbiota and immune system and suggest the possibility of improving neonatal health through the manipulation of maternal microbiota.

Pig weaning cause transient stress and reduction of feed intake and intestinal villi. Providing feed with solid texture could stimulate explorative interest, relieving weaning drawbacks and improving welfare. Feed blocks (0.8 kg each) were produced and 8 formulas were preliminarily tested for consumption on a pig farm (ingredients: wheat by-products, dried milk whey, calcium carbonate, oil, molasses). Feed consumption and growth within 3 days after weaning were assessed on penned litters fed the normal feed ad libitum, and one of the 3 best block formulas or a control (wooden pieces). Block consumption was relevant, but growth and carcase quality on a sub-sample reared to commercial maturation were not changed. These formulations were also tested against control on 72 weaned pigs on which behaviour related to feeding and social activities was evaluated by means of surveys using cameras. After 4 days, pigs were slaughtered and the small intestines were sampled for mucosa morphology. In general, block consumption was additive with the consumption of normal feed. Growth was not affected. One formulation (major ingredients: wheat middlings, cane molasses, milk whey and coconut oil) increased the mucosal surface area of the intestinal villi by 7.9% (p p Ingredient composition may influence the disappearance of the feed block, as a variable combination of disaggregation and intake by the piglets.The formulation based on wheat middlings, cane molasses, milk whey and coconut oil may improve growth and favour better welfareBlock supplementation may increase the absorbent surface of the proximal jejunum. Ingredient composition may influence the disappearance of the feed block, as a variable combination of disaggregation and intake by the piglets. The formulation based on wheat middlings, cane molasses, milk whey and coconut oil may improve growth and favour better welfare Block supplementation may increase the absorbent surface of the proximal jejunum.

The pathogenicity of Enterotoxigenic Escherichia coli (ETEC) expressing F4ac (K88) fimbrial adhesin and its adhesion to the pig jejunum surface is genetically controlled by the presence of a specific receptor glycoprotein on brush borders of the epithelium. We firstly screened the degree of inhibition obtained with ten galactose-recognising lectins – respectively from Abrus precatorius (APA-I); Coregonus maraena; Euphorbia characias; Crotalaria juncea; Sambucus ebulus (SEL1d); Sambucus nigra (SNAV); Coregonus peled; Momordica charantia (MCL); Trychosanthes kirilowii; Adenia racemosa –, by an ELISA test based on intestinal brush border sensitive for E. coli F4ac adhesion and ETEC F4ac fimbrial antigen. Three lectins significantly inhibited the fimbrial adhesion and the highest value was obtained by Crotalaria juncea lectin (p

The aim of the study was to characterize the soluble metabolomics profile of defatted colostrum of sows at different parity number (PA) and to correlate the metabolomics profile with the Brix percentage estimate of colostrum immunoglobulin G (IgG) and sow productive traits. A total of 96 Meidam (crossbreed Large White &times, Meishan) sows of PA from 1&ndash, 4 (PA1: 28, PA2:26, PA3:12, PA4:26) were included, and their productive traits were recorded at 10 days post-farrowing. Colostrum IgG was quantified using a Brix refractometer, and metabolomics profile was assessed using 1H-NMR spectroscopy. Sows&rsquo, PA slightly influenced the metabolomics profile of colostrum. lactose and glycine were higher in PA1 compared with PA4 (p 0.05) and N-acetylglucosamine (GlcNAc) tended to be higher in PA2 than PA3 and PA4 (p &lt, 0.10). The Brix percentage of IgG was negatively associated with lactose and positively with creatine, myo-inositol, and O-phosphocholine (p &lt, 0.05). Taurine was positively related to litter weight at birth. GlcNAc and myo-inositol were linked to piglet mortality at day 10 with a negative and positive trend, respectively. In conclusion, colostrum of gilts and multiparous sows had a similar metabolomics profile. Specific metabolites contributed to explanation of the variability in colostrum Brix percentage estimate of IgG concentration and the sows&rsquo, productive performance.

Background The stomach is an underestimated key interface between the ingesta and the digestive system, affecting the digestion and playing an important role in several endocrine functions. The quality of starter microbiota and the early life feeding of medium chain triglycerides may affect porcine gastric maturation. Two trials (T1, T2) were carried out on 12 and 24 cesarean-delivered piglets (birth, d0), divided over two microbiota treatments, but slaughtered and sampled at two or three weeks of age, respectively. All piglets were fed orally: sow serum (T1) or pasteurized sow colostrum (T2) on d0; simple starter microbiota (Lactobacillus amylovorus, Clostridium glycolicum and Parabacteroides spp.) (d1-d3); complex microbiota inoculum (sow diluted feces, CA) or a placebo (simple association, SA) (d3-d4) and milk replacer ad libitum (d0-d4). The The T1 piglets and half of the T2 piglets were then fed a moist diet (CTRL); the remaining half of the T2 piglets were fed the CTRL diet fortified with medium chain triglycerides and 7% coconut oil (MCT). Total mRNA from the oxyntic mucosa was analyzed using Affymetrix©Porcine Gene array strips. Exploratory functional analysis of the resulting values was carried out using Gene Set Enrichment Analysis. Results Complex microbiota upregulated 11 gene sets in piglets of each age group vs. SA. Of these sets, 6 were upregulated at both ages, including the set of gene markers of oxyntic mucosa. In comparison with the piglets receiving SA, the CA enriched the genes in the sets related to interferon response when the CTRL diet was given while the same sets were impoverished by CA with the MCT diet. Conclusions Early colonization with a complex starter microbiota promoted the functional maturation of the oxyntic mucosa in an age-dependent manner. The dietary fatty acid source may have affected the recruitment and the maturation of the immune cells, particularly when the piglets were early associated with a simplified starter microbiota.

The GPR120 gene (also known as FFAR4 or O3FAR1) encodes for a functional omega-3 fatty acid receptor/sensor that mediates potent insulin sensitizing effects by repressing macrophage-induced tissue inflammation. For its functional role, GPR120 could be considered a potential target gene in animal nutrigenetics. In this work we resequenced the porcine GPR120 gene by high throughput Ion Torrent semiconductor sequencing of amplified fragments obtained from 8 DNA pools derived, on the whole, from 153 pigs of different breeds/populations (two Italian Large White pools, Italian Duroc, Italian Landrace, Casertana, Pietrain, Meishan, and wild boars). Three single nucleotide polymorphisms (SNPs), two synonymous substitutions and one in the putative 3'-untranslated region (g.114765469C > T), were identified and their allele frequencies were estimated by sequencing reads count. The g.114765469C > T SNP was also genotyped by PCR-RFLP confirming estimated frequency in Italian Large White pools. Then, this SNP was analyzed in two Italian Large White cohorts using a selective genotyping approach based on extreme and divergent pigs for back fat thickness (BFT) estimated breeding value (EBV) and average daily gain (ADG) EBV. Significant differences of allele and genotype frequencies distribution was observed between the extreme ADG-EBV groups (P < 0.001) whereas this marker was not associated with BFT-EBV.