Intensive farming may involve the use of diets, environments or management practices that impose physiological and psychological stressors on the animals. In particular, early weaning is nowadays a ...common practice to increase the productive yield of pig farms. Still, it is considered one of the most critical periods in swine production, where piglet performance can be seriously affected and where they are predisposed to the overgrowth of opportunistic pathogens. Pig producers nowadays face the challenge to overcome this situation in a context of increasing restrictions on the use of antibiotics in animal production. Great efforts are being made to find strategies to help piglets overcome the challenges of early weaning. Among them, a nutritional strategy that has received increasing attention in the last few years is the use of probiotics. It has been extensively documented that probiotics can reduce digestive disorders and improve productive parameters. Still, research in probiotics so far has also been characterized as being inconsistent and with low reproducibility from farm to farm. Scientific literature related to probiotic effects against gastrointestinal pathogens will be critically examined in this review. Moreover, the actual practical approach when using probiotics in these animals, and potential strategies to increase consistency in probiotic effects, will be discussed. Thus, considering the boost in probiotic research observed in recent years, this paper aims to provide a much-needed, in-depth review of the scientific data published to-date. Furthermore, it aims to be useful to swine nutritionists, researchers and the additive industry to critically consider their approach when developing or using probiotic strategies in weaning piglets.
Weaning is a critical period in the life of pigs with repercussions on their health and welfare and on the economy of the swine industry. This study aimed to assess the effect of the commercial early ...weaning on gut microbiota, intestinal gene expression and serum metabolomic response via an integrated-omic approach combining 16S rRNA gene sequencing, the OpenArray gene expression technology and
H-NMR spectroscopy. Fourteen piglets from different litters were sampled for blood, jejunum tissue and caecal content two days before (- 2d), and three days after (+ 3d) weaning. A clearly differential ordination of caecal microbiota was observed. Higher abundances of Roseburia, Ruminococcus, Coprococcus, Dorea and Lachnospira genera in weaned piglets compared to prior to weaning showed the quick microbial changes of the piglets' gut microbiota. Downregulation of OCLN, CLDN4, MUC2, MUC13, SLC15A1 and SLC13A1 genes, also evidenced the negative impact of weaning on gut barrier and digestive functions. Metabolomic approach pinpointed significant decreases in choline, LDL, triglycerides, fatty acids, alanine and isoleucine and increases in 3-hydroxybutyrate after weaning. Moreover, the correlation between microbiota and metabolome datasets revealed the existence of metabolic clusters interrelated to different bacterial clusters. Our results demonstrate the impact of weaning stress on the piglet and give insights regarding the associations between gut microbiota and the animal gene activity and metabolic response.
The aim of this study was to determine the possible impact of early socialization and an enriched neonatal environment to improve adaptation of piglets to weaning. We hypothesized that changes in the ...microbiota colonization process and in their metabolic response and intestinal functionality could help the animals face weaning stress. A total of 48 sows and their litters were allotted into a control (CTR) or an enriched treatment (ENR), in which piglets from two adjacent pens were combined and enriched with toys. The pattern of caecal microbial colonization, the jejunal gene expression, the serum metabolome and the intestinal physiology of the piglets were assessed before (-2 d) and after weaning (+ 3d). A differential ordination of caecal microbiota was observed after weaning. Serum metabolome suggested a reduced energetic metabolism in ENR animals, as evidenced by shifts in triglycerides and fatty acids, VLDL/LDL and creatine regions. The TLR2 gene showed to be downregulated in the jejunum of ENR pigs after weaning. The integration of gene expression, metabolome and microbiota datasets confirmed that differences between barren and enriched neonatal environments were evident only after weaning. Our results suggest that improvements in adaptation to weaning could be mediated by a better response to the post-weaning stress.
The potential of a prebiotic oligosaccharide lactulose, a probiotic strain of Lactobacillus plantarum, or their synbiotic combination to control postweaning colibacillosis in pigs was evaluated using ...an enterotoxigenic Escherichia coli (ETEC) K88 oral challenge. Seventy-two weanlings were fed four diets: a control diet (CTR), that diet supplemented with L. plantarum (2 × 10(10) CFU · day(-1)) (LPN), that diet supplemented with 10 g · kg(-1) lactulose (LAC), or a combination of the two treatments (SYN). After 7 days, the pigs were orally challenged. Six pigs per treatment were euthanized on days 6 and 10 postchallenge (PC). Inclusion of lactulose improved the average daily gain (ADG) (P < 0.05) and increased lactobacilli (P < 0.05) and the percentage of butyric acid (P < 0.02) in the colon. An increase in the ileum villous height (P < 0.05) and a reduction of the pig major acute-phase protein (Pig-MAP) in serum (P < 0.01) were observed also. The inclusion of the probiotic increased numbers of L. plantarum bacteria in the ileum and colon (P < 0.05) and in the total lactobacilli in the colon and showed a trend to reduce diarrhea (P = 0.09). The concentrations of ammonia in ileal and colonic digesta were decreased (P < 0.05), and the villous height (P < 0.01) and number of ileal goblet cells (P < 0.05) increased, at day 10 PC. A decrease in plasmatic tumor necrosis factor alpha (TNF-α) (P < 0.01) was also seen. The positive effects of the two additives were combined in the SYN treatment, resulting in a complementary synbiotic with potential to be used to control postweaning colibacillosis.
An experiment was designed to evaluate the effects of 3 different additives on the gastrointestinal microbiota of early-weaned pigs. Early-weaned (18 to 22 d; n = 32) pigs (6.0 ± 0.10 kg of BW) from ...8 litters were randomly distributed into 8 pens. Each pen was assigned 1 of 4 dietary treatments: a prestarter or control diet, the control diet with 0.04% avilamycin (AB), with 0.3% sodium butyrate, or with 0.03% plant extract mixture (XT; standardized mixture with 5% (wt/wt) carvacrol extracted from Origanum spp., 3% cinnamaldehyde extracted from Cinnamonum spp., and 2% capsicum oleoresin from Capsicum annum). At the end of the experimental period, 8 pigs per treatment were killed, and samples of their intestinal content were taken. The total bacterial load along the gastrointestinal tract (GIT; stomach, jejunum, cecum, and distal colon) and the lactobacilli and enterobacteria in the jejunum and cecum were measured by quantitative PCR. The total microbial counts along the GIT did not differ among the diets, but there was an increase in the lactobacilli:enterobacteria ratio in the cecum of the piglets on the XT diet (P = 0.003). Restriction fragment length polymorphism of the PCR-amplified V3, V4, and V5 regions of the 16S rDNA gene showed changes in the structure of the microbial community in the jejunum. Dendrograms grouped animals by diets; control with 0.3% sodium butyrate was the treatment that promoted the biggest changes in the microbial ecosystem, followed by AB and then XT. Biodiversity increased when using additives compared with the control diet (P = 0.002). Microbial metabolic activity along the hindgut was studied using the concentration of purine bases and carbohydrase activities. Different patterns for purine bases were observed between diets (diet x intestinal section, P = 0.01). The control diet reached a maximum purine base concentration at the end of the colon, whereas that of the AB diet was reached at the cecum. We could not detect any cellulase or xylanase activities in animals of this age. Appreciable amylase and amylopectinase activities were found, but they did not differ between diets. The results suggest that the effects of avilamycin, butyrate, or the plant extract would not be related to a reduction in the number of total bacteria inhabiting different sections of the GIT but rather to changes in the ecological structure and metabolic activity of the microbial community.
Enterotoxigenic Escherichia coli (ETEC) K88 is the most prevalent enteropathogen in weaned piglets, with the ability to express fimbria F4 and specifically attach to intestinal receptors in the young ...piglet. The prevention of ETEC K88 adhesion to the epithelium by interfering in this fimbria-receptor recognition provides an alternative approach to prevent the initial stage of disease. The aim of this study is to screen, among different feed ingredients (FI), their ability to reduce ETEC K88 attachment to the porcine intestinal epithelial cell-line (IPEC-J2). The selected FI consisted of products of a vegetable or dairy origin, and microbial by-products, which could be suitable to be included in piglet's diet. Incubation of a mixture of each FI extract with the bacteria on IPEC-J2 monolayer was allowed. After washing with PBS to remove the non-adhered bacteria, the culture medium was added to grow the adhered bacteria and, simultaneously, to keep the cells alive. Then, the bacterial growth was monitored in a spectrophotometer reader for 12h. Casein glycomacropeptide (CGMP), locust bean (LB), exopolysaccharide (EPS) and wheat bran (WB) reduced the number of attached ETEC K88 to IPEC-J2, but no anti-adhesive effect was found for soybean hulls, sugar-beet pulp, locust gum, fructooligosaccharides, inulin, mushroom, mannanoligosaccharides or the fermented product from Aspergillus oryzae. The lineal analysis of dose responses demonstrated lineal activity (P<0.0001) for CGMP, LB, EPS and WB. These in vitro results suggest CGMP, LB, EPS and WB as good candidates to be included in piglet's diet with supported functional activity against colibacillosis.
We evaluated the effects of 3 additives, sodium butyrate (AC), avilamycin (AB), and a combination of plant extracts (XT), on the productive performance and the intestinal environment of the ...early-weaned pig. The XT was a standardized mixture with 5% (wt/wt) carvacrol (from Origanum spp.), 3% cinnamaldehyde (from Cinnamonum spp.), and 2% capsicum oleoresin (from Capsicum annum). Pigs (n = 32) weaned at 18 to 22 d of age with an initial BW of 6.0 ± 0.10 kg were allocated to 8 pens that, in turn, were allocated to 4 treatments. The treatments included a basal diet (CT) or the basal diet supplemented with 0.3% of AC, 0.04% of AB, or 0.03% of XT. Productive performance was determined during the initial 14 d postweaning. On d 19 and 21 of the experiment, the pigs were killed to allow collection of digesta and intestinal tissue to evaluate variables indicative of aspects of the gastrointestinal environment. Treatments AB and AC improved G:F (P = 0.012 and 0.003, respectively) compared with the CT. Butyrate included in the diet was only detected in the stomach but not in cranial jejunum. When compared with CT, AC produced a lower ileal starch digestibility (P = 0.002) and a lower whole-tract OM and starch digestibility (P = 0.001 and 0.003, respectively), related to a lower VFA concentration in the cranial colon (P = 0.082) and a numerically reduced branched VFA percentage in the rectum. The AB treatment diminished propionate production in caudal colon (P = 0.002) and rectum (P = 0.012) compared with CT. The AC group exhibited deeper crypt depth in the jejunum without variations in villus height compared with CT (P = 0.042). The AC and AB groups also increased goblet cell presence in the colon (P = 0.001 and 0.032, respectively). On the other hand, AB and XT diminished intraepithelial lymphocytes in the jejunum (P = 0.003 and 0.034, respectively). The XT increased lymphocyte presence in the colon (P = 0.003). These results show the important influence of AB and AC on productive performance and on pig gut dynamics. The intestinal modifications observed for AB and AC compared with CT suggest distinct modes of action for each additive.
•Inclusion of 1.6% of limestone reduced body weight, average daily gain and feed conversion ratio, but not feed intake.•Inclusion of 1.6% of limestone upregulated the expression of genes related to ...the inflammatory response in the jejunum.•Inclusion of 1.6% of limestone increased beta-diversity and Bacteroides genera in colon.
Plant ingredients contain low Ca levels and 1/3 may be bound to P phytate. Therefore, Ca has to be supplemented with animal or mineral sources, such as limestone or calcium phosphates. However, growth performance and feed intake has been reported by other authors to decrease linearly with increasing dietary Ca. To better understand the possible mechanisms underlying detrimental effect of including limestone in weanling diets, two experimental trials were performed to evaluate its possible impact on growth performance, colonic microbiota and, jejunal gene expression. In Trial 1, a total of 240 piglets (26 ± 2 d old, average body weight (BW) = 7.7 ± 1.04 kg) were distributed into 3 dietary treatments: 0% limestone, 0.8% limestone, and 1.6% limestone to provide 0.6, 0.9 and 1.2% total Ca, respectively. Feed intake and individual BW were registered during the pre-starter phase (d 0–14 post-weaning). Pigs fed diet with 1.6% of limestone (1.2% of total Ca) had lower BW, average daily gain (ADG), and gain to feed ratio (G:F) compared with pigs fed diets with 0.8 and 0% limestone (0.9 and 0.6% total Ca, respectively; P < 0.05). In Trial 2, a total of 18 pigs (28 ± 0 d old, average BW = 7.2 ± 0.24 kg) were allocated individually in cages and assigned to 2 dietary treatments: 0% limestone and 1.4% limestone to provide 0.6 and 1.2% total Ca, respectively. Piglets were fed for 14 d and then euthanized to obtain jejunum tissue for gene expression and colon digesta for 16SRNA gene microbiota analyses. Pigs fed 1.4% limestone (1.2% of total Ca) showed a higher beta-diversity and an increase on the Bacteroides genera in colon (P < 0.05). These animals also showed an up regulation of gene sets related to the cell cycle regulation, DNA and RNA transcription and inflammatory response in the jejunum. In conclusion, high levels of limestone inclusion in weaning diets, decreases growth performance without affecting feed intake in weaned pigs. Also, addition of limestone to diets for 14 d after weaning can upregulate the expression of genes related to the inflammatory response, and enlarge colonic beta-diversity with an increased Bacteroides genera.
The inclusion of genetically modified (GM) plants in the human diet has raised concerns about the possible transfer of transgenes from GM plants to intestinal microflora and enterocytes. The ...persistence in the human gut of DNA from dietary GM plants is unknown. Here we study the survival of the transgene epsps from GM soya in the small intestine of human ileostomists (i.e., individuals in which the terminal ileum is resected and digesta are diverted from the body via a stoma to a colostomy bag). The amount of transgene that survived passage through the small bowel varied among individuals, with a maximum of 3.7% recovered at the stoma of one individual. The transgene did not survive passage through the intact gastrointestinal tract of human subjects fed GM soya. Three of seven ileostomists showed evidence of low-frequency gene transfer from GM soya to the microflora of the small bowel before their involvement in these experiments. As this low level of epsps in the intestinal microflora did not increase after consumption of the meal containing GM soya, we conclude that gene transfer did not occur during the feeding experiment.
The present study evaluated
in vitro the effect of technological processes on four cereal grains: maize, wheat, barley and oats and one legume grain: green seeded pea. Processing included extrusion, ...cooking and grinding (0.8 and 3.0
mm opening), and the parameters evaluated were starch hydrolysis, content of soluble non-starch polysaccharides (NSP) and physicochemical properties: solubility of dry matter (DM), viscosity of the supernatant and water retention capacity (WRC).
All samples were submitted to an
in vitro digestion that consisted of two steps, gastric (G; pepsin; pH 2; 4
h) and small intestine (SI; pancreatin; pH 7; 4
h) simulation. Prior to incubation, particle size was evaluated in all samples by hand sieving and calculated on a weight basis. Total starch and total NSP analyses were performed on raw samples (0.8
mm). Hydrolysis of starch was evaluated by measuring glucose in the supernatant during the course of SI incubation. Soluble NSP were evaluated in the supernatant at the end of SI incubation. Physicochemical properties were evaluated after G and SI incubation.
In raw ingredients (0.8
mm) the hydrolysis of starch at the end of SI incubation (ratio between glucose released in the sample and glucose released in the sample gelatinized) was lower in peas (0.45) than in cereal grains (0.69 in maize; 0.95 in wheat; 0.82 in barley and 0.92 in oats). The highest amounts of soluble NSP were found in barley (19.8
g/kg DM) and oats (13.8
g/kg DM), and were associated with higher viscosities. The WRC in raw samples was positively related to the amount of total NSP after both G (
r
2
=
0.847, P<0.001 for 0.8
mm samples) and SI incubation (
r
2
=
0.773, P<0.001).
Technological processing increased starch hydrolysis and solubilization of NSP in all ingredients. The extent of the effect was dependent on the ingredient as well as on the severity of the technological process. Component monosaccharides of soluble NSP were differently affected, while glucose was increased by all the processes, arabinose and xylose required the more extreme conditions of extrusion. Contrary to raw samples, WRC of heat treated ingredients showed a negative relation with total NSP content of the sample after G incubation, which turned out to be a positive relation after SI incubation.
It is concluded that technological processing of ingredients promotes higher starch hydrolysis, increases in the amount of soluble NSP and modifications in the physicochemical properties depending on the nature of the feed ingredients.