•Functional amino acids play a valuable role in pigs’ gut barrier and immunity.•Functional amino acids have signalling molecule activities.•Arginine, Glutamate and Glutamine effectively promote gut ...morphology and immunity.•Functional amino acids are among the strategies for reducing in-feed antimicrobials.•Feeding combinations in amino acids could be a strategy to promote gut health.
The need to reduce the use of antibiotics and zinc oxide at the pharmacological level, while preserving the performance of postweaning piglets, involves finding adequate nutritional strategies which, coupled with other preventive strategies, act to improve the sustainability of the piglet-rearing system. Amino acids (AAs) are the building blocks of proteins; however, they also have many other functions within the body. AA supplementation, above the suggested nutritional requirement for piglets, has been investigated in the diets of postweaning piglets to limit the detrimental consequences occurring during this stressful period. A systematic review was carried out to summarise the effects of AAs on gut barrier function and immunity, two of the parameters contributing to gut health. An initial manual literature search was completed using an organised search strategy on PubMed, utilising the search term “<amino acid> AND <parameter related to intestinal health>”. These searches yielded 302 articles (published before October 2021); 59 were selected. Based on the method for extracting data (synthesis of evidence), this review showed that L-Arginine, L-Glutamine and L-Glutamate are important functional AAs playing major roles in gut morphology and immune functions. Additional benefits of AA supplementation, refereed to a supplementation above the suggested nutritional requirement for piglets, could also be observed; however, data are needed to provide consistent evidence. Taken together, this review showed that supplementation with AAs during the weaning phase supported a plethora of the physiological functions of piglets. In addition, the data reported confirmed that each amino acid targets different parameters related to gut health, suggesting the existence of potential synergies among them.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Dietary resistant starch (RS) may have prebiotic properties but its effects on fermentation and the microbial population are inconsistent. This meta-analysis aimed to quantify the relationship ...between RS type 2 (RS2) and intestinal short-chain fatty acids (SCFA) and pH as well as certain key bacterial taxa for intestinal health in pigs. From the 24 included articles with sufficient information about the animal, and dietary and physiological measurements published between 2000 and 2017, individual sub-data sets for fermentation metabolites, pH, bacterial abundances and apparent total tract digestibility were built and used to parameterize prediction models on the effect of RS2, accounting for inter- and intra-study variability. In addition, the effect of pig’s BW at the start of the experiment and duration of the experimental period on response variables were also evaluated using backward elimination analysis. Dietary RS levels ranged from 0% to 78.0% RS, with median and mean RS levels of 28.8% and 23.0%, respectively. Negative relationships could be established between dietary RS and pH in the large intestine (P<0.05), with a stronger effect in the mid and distal colon, and feces (R2=0.64 to 0.81; P<0.001). A dietary level of 15% RS would lower the pH in the proximal, mid-, distal colon and feces by 0.2, 0.6, 0.4 and 0.6 units, respectively. Increasing RS levels, however, did not affect SCFA concentrations in the hindgut, but enhanced the molar proportion of propionate in mid-colon and reduced those of acetate in mid-colon and of butyrate in mid- and distal colon (R2=0.46 to 0.52; P<0.05). Backward elimination indicated an age-related decrease in mid-colonic propionate proportion and increase in mid- and distal colonic butyrate proportion (P<0.05), thereby modulating RS2 effects. In feces, increasing RS levels promoted fecal lactobacilli (R2=0.46; P<0.01) and bifidobacteria (R2=0.57; P<0.01), whereby the slope showed the need for a minimal RS level of 10% for a 0.5 log unit-increase in their abundance. Best-fit equations further supported that a longer experimental period increased fecal lactobacilli but decreased fecal bifidobacteria (P<0.05). In conclusion, dietary RS2 seems to effectively decrease digesta pH throughout the large intestine and increase lactic acid-producing bacteria in feces of pigs which may limit the growth of opportunistic pathogens in the hindgut. To achieve these physiologically relevant changes, dietary RS should surpass 10% to 15%.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
In the last decades, gut microbiota and its role in mammal host development and health have been increasingly investigated. Metabolites produced by gut microbiota can affect intestinal homeostasis ...and immune system maturity and activation, and in turn, they can influence the health and growth performance of livestock. Therefore, a better understanding of the functional metabolic capability of the gut microbiota would be appreciated by the scientific community. In this study, the Biolog
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Ecoplates technology was applied for studying the metabolic potential of the aerotolerant microbial community of pig fecal samples, evaluating the interference of different storage conditions and cell concentrations. The length of time for which a fecal sample maintained detectable and unchanged microbial metabolic activity was also investigated. Two assays aimed to evaluate differences in the metabolic activities between fresh and snap-frozen fecal samples at different dilutions and at different lengths of times of preservation at −80°C were carried out. The biodiversity and the predicted functionality of the entire bacterial community through a targeted metagenomic approach were also explored. The results highlighted that snap freezing of fecal samples preserved the metabolic activity of the microbial community when compared to fresh feces. Sample storage at −80 °C did not significantly affect the metabolic activity of the microbial community, which was stable for 150 days. Furthermore, the highest metabolic activity was detected with 1:2 to 1:5 dilutions of the stock suspension. Biolog
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Ecoplates technology is a rapid and useful method to explore microbial communities’ metabolism in animal fecal samples contributing to investigate host animal physiology.
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Freezing of samples can preserve the functional activity of the aerotolerant microbial community for 150 days.
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The concentration of microbial cells strongly influences metabolic activity detection.
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Sequencing coupled with the Biolog
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Ecoplates could be a strategy to evaluate the metabolic potential of the microbiota of the fecal sample.
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CEKLJ, DOBA, EMUNI, FZAB, GEOZS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
•E. coli oral vaccine can modulate gut microbiome and in immunity of piglets.•Feed intake and gut integrity were promoted by E. coli oral vaccine.•E. coli vaccine reduced the diarrhoea in the first ...14 days postweaning.•Vaccination showed a probiotic-like effect by modulating gut microbiota.•E. coli oral vaccine may represent a possible tool to improve piglets’ resilience.
Oral live vaccines stimulate host immunity, but they could also affect intestinal mucosa development and gut microbiota of piglets during the postweaning. The aim of this study was to determine the effect of an oral vaccine against Escherichia coli F4 and F18 (Coliprotec F4/F18®), on gut functionality and integrity, growth performance and health status of postweaning piglets. A total of 96 weaned piglets (23.30 ± 1.85 days of age; 7334 ± 1039 g BW) were divided into two groups (16 replicates/group; three piglets/replicate) as follows: (1) Control (CO), fed a standard diet (prestarter up to 14 days, then starter feed); (2) Treated (TRT): as CO but vaccinated with Coliprotec F4/F18® at weaning (day 0). Piglets were weighed at day 0 and weekly until day 35. Individual faecal score was recorded daily. Piglets were sacrificed at days 10 (1/3 of total) and 35 (2/3). Samples of jejunum mucosa and of cecum content were collected for morphometric, immunohistochemistry analysis and for microbiota profile analysis, respectively. Data were fitted using a linear model including treatment, class of starting BW as fixed factors and litter as random factor. From days 0 to 7, piglets from the TRT group tended to have a higher average daily gain (+22.6%, P = 0.08) and average daily feed intake compared to the CO group (+13.2%, P = 0.022). Gain to feed ratio was lower in the TRT group from days 14 to 35 (−6.6%, P = 0.011). From days 7 to 14, the TRT group had a higher diarrhoea index (−199%, P < 0.001). Crypt depth was higher in the CO group (+10.9%, P = 0.04) at day 10, but not at day 35. Jejunal expression of Claudin-4 (probability of having a score = 3) was higher in the TRT group at day 10 (CO = 1.50% vs TRT = 2.69%, P < 0.0001) and day 35 (CO = 1.29% vs TRT = 1.92%, P = 0.012). Oral vaccine affected beta diversity at day 10 (P = 0.040; R2 = 0.05) and increased the abundance of specific taxa and genera in the cecum at day 10, including Prevotella (lg2FC = 23.2, FDR < 0.001). The results showed how an Escherichia coli-based vaccine supplied to weaned pigs can promote gut health by controlling symptoms of the postweaning perturbation in the first 2 weeks postweaning. In addition, the vaccine strains showed a probiotic-like effect by modulating gut microbiota favouring the establishment of beneficial bacteria, and by promoting gut barrier integrity.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Porcine colostrum lipid classes and fatty acids (FA) were characterized in 6 pools (from 69 samples) from 3 sow breeds (Italian Large White, Italian Landrace, and Italian Duroc) and different parity ...orders (only Large White). Triacylglycerols (TAG; 94.44 expressed as g/100 g of fat) were the most abundant lipid class, followed by diacylglycerols (DAG; 3.36 g/100 g of fat), free fatty acids (FFA; 0.98 g/100 g of fat), and cholesterol (0.84 g/100 g of fat). The main FAs found in swine colostrum were palmitic (27.29%, expressed as g/100 g of total FA), oleic (28.81%), and linoleic (23.39%) acids. Both the breed of sow and parity order affected the FA and lipid composition. The results suggest that the FA composition of swine colostrum is similar to that of human colostrum and could represent a new source of nutrients for human infants, after further assessment of hygienic and quality aspects. The swine model could be an opportunity for a better understanding of colostrum effects on newborns.
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IJS, KILJ, NUK, PNG, UL, UM, UPUK
The ability of live yeasts to modulate pig intestinal cell signals in response to infection with Escherichia coli F4ac (ETEC) has not been studied in-depth. The aim of this trial was to evaluate the ...effect of Saccharomyces cerevisiae CNCM I-4407 (Sc), supplied at different times, on the transcriptome profile of the jejunal mucosa of pigs 24 h after infection with ETEC. In total, 20 piglets selected to be ETEC-susceptible were weaned at 24 days of age (day 0) and allotted by litter to one of following groups: control (CO), CO+colistin (AB), CO+5×1010 colony-forming unit (CFU) Sc/kg feed, from day 0 (PR) and CO+5×1010 CFU Sc/kg feed from day 7 (CM). On day 7, the pigs were orally challenged with ETEC and were slaughtered 24 h later after blood sampling for haptoglobin (Hp) and C-reactive protein (CRP) determination. The jejunal mucosa was sampled (1) for morphometry; (2) for quantification of proliferation, apoptosis and zonula occludens (ZO-1); (3) to carry out the microarray analysis. A functional analysis was carried out using Gene Set Enrichment Analysis. The normalized enrichment score (NES) was calculated for each gene set, and statistical significance was defined when the False Discovery Rate % was <25 and P-values of NES were <0.05. The blood concentration of CRP and Hp, and the score for ZO-1 integrity on the jejunal villi did not differ between groups. The intestinal crypts were deeper in the AB (P=0.05) and the yeast groups (P<0.05) than in the CO group. Antibiotic treatment increased the number of mitotic cells in intestinal villi as compared with the control group (P<0.05). The PR group tended to increase the mitotic cells in villi and crypts and tended to reduce the cells in apoptosis as compared with the CM group. The transcriptome profiles of the AB and PR groups were similar. In both groups, the gene sets involved in mitosis and in mitochondria development ranked the highest, whereas in the CO group, the gene sets related to cell junction and anion channels were affected. In the CM group, the gene sets linked to the metabolic process, and transcription ranked the highest; a gene set linked with a negative effect on growth was also affected. In conclusion, the constant supplementation in the feed with the strain of yeast tested was effective in counteracting the detrimental effect of ETEC infection in susceptible pigs limits the early activation of the gene sets related to the impairment of the jejunal mucosa.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The EU ban on in-feed antibiotics has stimulated research on weaning diets as a way of reducing post-weaning gut disorders and growth check in pigs. Many bioactive components have been investigated ...but only few have shown to be effective. Amongst these, organic acids (OA) have been shown to exert a bactericidal action mediated by non-dissociated OA, by lowering gastric pH, increasing gut and pancreas enzyme secretion and improving gut wall morphology. It has been postulated that they may also enhance non-specific immune responses and improve disease resistance. In contrast, relatively little attention has been paid to the impact of OA on the stomach but recent data show they can differently affect gastric histology, acid secretion and gastric emptying. Butyrate and precursors of butyric acid have received special attention and although promising results have been obtained, their effects are dependent upon the dose, treatment duration, initial age of piglets, gastrointestinal site and other factors. The amino acids (AA) like glutamine, tryptophan and arginine are supportive in improving digestion, absorption and retention of nutrients by affecting tissue anabolism, stress and (or) immunity. Glutamine, cysteine and threonine are important for maintaining mucin and permeability of intestinal barrier function. Spray-dried plasma (SDP) positively affects gut morphology, inflammation and reduces acquired specific immune responses via specific and a-specific influences of immunoglobulins and other bioactive components. Effects are more pronounced in early-weaned piglets and under poorer health conditions. Little interaction between plasma protein and antibiotics has been found, suggesting distinct modes of action and additive effects. Bovine colostrum may act more or less similarly to SDP. The composition of essential oils is highly variable, depending on environmental and climatic conditions and distillation methods. These oils differ widely in their antimicrobial activity in vitro and some components of weaning diets may decrease their activity. Results in young pigs are highly variable depending upon the product and doses used. These studies suggest that relatively high concentrations of essential oils are needed for beneficial effects to be observed and it has been assumed that these plant extracts mimic most of the effects of antibiotics active on gut physiology, microbiology and immunology. Often, bioactive substances protective to the gut also stimulate feed intake and growth performance. New insights on the effects of selected OA and AA, protein sources (especially SDP, bovine colostrum) and plant extracts with anti-bacterial activities on the gut are reported in this review.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Scarce is knowledge on the process regulating the development of acid secretion, orexigenic signaling, and chemosensing in the stomach of young pigs. Changes of early microbial encounters by suckling ...pigs can interact with the gut maturation, by the induction of different molecular signaling. Our goal was to assess if the age of offspring and the maternal environment, influenced by sow antibiotic treatment peripartum, could affect gastric morphology and the expression of genes involved in the control of hydrochloric secretion, feed intake, taste, and inflammation in offspring stomach.
84 pigs from sows fed a diet with amoxicillin (on -d10 to +d21 from farrowing, ANT) or without (CON) were sacrificed at d14, d21, d28 (weaning) or d42. Samples of oxyntic (OXY), pyloric (PY) and cardiac mucosae close to OXY were collected and parietal and enteroendocrine cells (EECs) were counted. Relative gene expression of a set of 11 key genes (ATP4A, SSTR2, GAST, GHRL, MBOAT4, PCSK1, GNAT1, TAS1R1, TAS1R3, IL8 and TNF) was assessed by qRT-PCR. In addition, 40 offspring obtained from the same ANT and CON sows were offered a normal or a fat-enriched diet for 4 weeks between 140 and 169 d of age, and then OXY and PY were sampled.
The number of parietal and EECs increased with age (P < 0.001). ATP4A increased with age (within suckling, P = 0.043, post-weaning vs. suckling, P < 0.001), SSTR2 increased only after weaning (P < 0.001). In OXY, GHRL increased during suckling (P = 0.012), and post-weaning as a trend (P = 0.088). MBOAT4 tended to increase during suckling (P = 0.062). TAS1R1 increased from suckling to post-weaning period (P =0.001) and was lower in ANT offspring (P = 0.013). GNAT1 in PY was higher in ANT offspring (P = 0.041). Antibiotic treatment of sows peripartum increased expression of GHRL and MBOAT4 in OXY of growing-finishing offspring aged 5 months.
Data show that sensing for umami taste and ghrelin regulation can be affected by maternal environment, but the development of acid secretion, orexigenic signaling and taste perception in the stomach are mostly developmentally controlled.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Knowledge on porcine bitter and fat taste receptors and on their expression in gastrointestinal tract of pigs is scarce. We searched for the presence of porcine homologous sequences for 13 human ...transcripts of bitter and fat taste receptors in ENSEMBL and National Center for Biotechnology Information databases. For taste 2 receptor (TAS2R) 8, alignment was not observed; for TAS2R13 and TAS2R46 the porcine predicted sequence aligned with several other human bitter genes. For 7 genes for bitter taste (TAS2R1, TAS2R3, TAS2R7, TAS2R9, TAS2R10, TAS2R16, and TAS2R38) and for 3 genes for fat taste (GPR40, GPR43, and GPR120), a full homology for exon sequences was found and primers were designed by Primer3. These 7 genes were amplified with real-time PCR and verified on agarose gel in 5 gastrointestinal segments of weaned pigs: oxyntic (ST1), pyloric (ST2), and cardiac to oxyntic transition mucosa (ST3), jejunum (JEJ), and colon (COL). Suitability of mRNA was verified by amplifying RPL4 and HMBS2 genes. Each bitter taste gene was detectable on agarose gel in at least 1 subject of all the gastrointestinal segments except for TAS2R3 and TAS2R38 that were never detected in ST1 and COL, respectively. The inspection of bitter taste genes amplification curve indicated that the expression was in general very low. GPR43 and GPR120 were present in all segments from all pigs. Expression was not detected for GPR40. Data also indicate that colon is the preeminent tract where fat detection by GPR120 takes place (P < 0.001). The presence of gene expression for several chemosensing receptors for bitter and fat taste in different compartments of the stomach confirms that this organ should be considered a player for the early detection of bolus composition.
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