Dietary xylo-oligosaccharides and arabinoxylans improved growth efficiency by reducing gut epithelial cell turnover in broiler chickens.

BACKGROUND: One of the main roles of the intestinal mucosa is to protect against environmental hazards. Supplementation of xylo-oligosaccharides (XOS) is known to selectively stimulate the growth of beneficial intestinal bacteria and improve gut health and function in chickens. XOS may have an impact on the integrity of the intestinal epithelia where cell turnover is critical to maintain the compatibility between the digestive and barrier functions. The aim of the study was to evaluate the effect of XOS and an arabinoxylan-rich fraction (AXRF) supplementation on gut function and epithelial integrity in broiler chickens. METHODS: A total of 128 broiler chickens (Ross 308) were assigned into one of two different dietary treatments for a period of 42 d: 1) control diet consisting of a corn/soybean meal-based diet; or 2) a control diet supplemented with 0.5% XOS and 1% AXRF. Each treatment was randomly distributed across 8 pens (n = 8) with 8 chickens each. Feed intake and body weight were recorded weekly. On d 42, one male chicken per pen was selected based on average weight and euthanized, jejunum samples were collected for proteomics analysis. RESULTS: Dietary XOS/AXRF supplementation improved feed efficiency (P < 0.05) from d 1 to 42 compared to the control group. Proteomic analysis was used to understand the mechanism of improved efficiency uncovering 346 differentially abundant proteins (DAP) (Padj < 0.00001) in supplemented chickens compared to the non-supplemented group. In the jejunum, the DAP translated into decreased ATP production indicating lower energy expenditure by the tissue (e.g., inhibition of glycolysis and tricarboxylic acid cycle pathways). In addition, DAP were associated with decreased epithelial cell differentiation, and migration by reducing the actin polymerization pathway. Putting the two main pathways together, XOS/AXRF supplementation may decrease around 19% the energy required for the maintenance of the gastrointestinal tract. CONCLUSIONS: Dietary XOS/AXRF supplementation improved growth efficiency by reducing epithelial cell migration and differentiation (hence, turnover), actin polymerization, and consequently energy requirement for maintenance of the jejunum of broiler chickens.

Using in feed xylanase or stimbiotic to reduce the variability in corn nutritive value for broiler chickens.

This study investigated the effects of xylanase and stimbiotic (fiber fermentation enhancer) on the response of broiler chickens fed different corn varieties and determine correlations between variables of interest. Four corn genetic varieties were selected based on their range in nutrient composition. Diets containing 600 g/kg of each corn were supplemented with 0 or 100 g/ton of xylanase or stimbiotic. A total of 1,152 one-day-old male broiler chicks (Ross 308) were divided into 12 treatments, each with 8 pens and 12 birds per pen, for a 21-day study. On d 21, performance parameters were measured, and the ileal energy and organic matter (OM) digestibility and cecal xylanase activity determined. Stimbiotic supplementation improved mFCR compared with all other treatments. There was a treatment by corn variety interaction for body weight (BW), BW gain and coefficient of variation (CV) of BW (P ≤ 0.05). Birds fed corn Variety 1 (highest neutral dietary fiber, protein and soluble arabinoxylan content) supplemented with stimbiotic had the highest BW, while Variety 2 control diet had the lowest. The BW CV in corn Variety 2 was the highest, which improved with stimbiotic supplementation. The BW CV in corn Variety 1 responded better to stimbiotic than xylanase. There were no treatment differences on BW CV in corn Varieties 3 and 4. The lowest OM digestibility was observed in birds fed corn Variety 1 with xylanase, and the highest value was associated with corn Variety 3 with xylanase (highest total arabinoxylan). Xylanase and stimbiotic supplementation increased the endogenous xylanase activity regardless of the corn variety (P ≤ 0.05). Positive correlations between corn fiber contents and phytic acid and the arabinose:xylose ratio were seen, while nonstarch polysaccharide content was negatively correlated with apparent metabolizable energy. In conclusion, corn variety influenced nutrient digestibility and broiler chicken growth. The response to supplementation with xylanase or stimbiotic varied based on the nutritional profile of corn with regards to improvements in digestibility and performance in broiler chickens.

Interactive effects of a stimbiotic supplementation and wheat bran inclusion in corn- or wheat-based diets on growth performance, ileal digestibility, and expression of nutrient transporters of broilers chickens.

A 42-day experiment was conducted to investigate the interactive effects of a stimbiotic (STB) and wheat bran (WB) in broiler chickens receiving diets (DT) based on corn or wheat. A total of 960 Cobb 500 male broiler chicks at zero-day old were allocated to 64 pens with 8 treatments, 8 replicates per treatment, and 15 birds per replicate. The treatments were arranged in a randomized complete block design with 2 × 2 × 2 factorial, with the factors as diet (corn-soybean meal or wheat-soybean meal), STB (with or without), and WB (0 or 50 g/kg). Body weight gain (BWG), feed intake (FI), and mortality-corrected FCR data were collected for the starter (d 0-10), grower (d 10-28), and finisher (d 28-42) phases. Ileal digesta and jejunal tissue were collected on d 18 and 42. Data were analyzed as a 2 × 2 × 2 factorial using a mixed model of JMP. There was no significant 3-way interaction for growth performance and expression of nutrient transporters. There was a significant SB × WB for FCR (P < 0.05) in the grower phase. Stimbiotic supplementation and WB inclusion individually improved (P < 0.05) FCR regardless of cereal type, but this effect was not observed when the two were combined. In the overall phase (d 0-42), birds receiving corn-based diets had higher (P < 0.05) FI and BWG than those receiving wheat-based diets. There was a significant 3-way interaction for ileal digestible energy (IDE) (P < 0.05) and N digestibility (ND) on d 18 and 42 (P < 0.05). There was no effect of STB or WB in corn-based diets, whereas WB inclusion decreased IDE in wheat-based diets, but STB supplementation increased IDE with or without WB. In corn-based diets, WB inclusion increased ND, whereas ND was decreased in wheat-based diets. Supplementation with STB had no effect on ND, but STB increased ND in wheat-based diets with and without WB. Ileal DE was greater (P < 0.05) for wheat- than corn-based diets on d 42. Wheat bran inclusion increased ND in corn-based diets whereas, with STB supplementation, there was no difference in ND with or without WB. There was no difference in ND with or without STB or WB in wheat-based diets. There was a significant STB × DT on the expression of GLUT-1 (P < 0.05). In corn-based diets, STB produced an upward expression of GLUT-1, whereas in wheat-based diets, STB supplementation had no effect. On d 42, stimbiotic supplementation produced upward (P < 0.05) expression of SGLT-1. In conclusion, STB supplementation in wheat- or corn-based diets and with WB inclusion improved energy digestibility. On the other hand, WB inclusion in wheat-based diets decreased nutrient digestibility especially on d 18, thus making room for a positive response to STB supplementation in wheat-based diets compared to the marginal response to STB supplementation observed in corn-based diets.

Stimbiotic supplementation and xylose-rich carbohydrates modulate broiler's capacity to ferment fibre.

Stimbiotics are a new category of feed additives that can increase fibre fermentability by stimulating fibre-degrading microbiota in the gut. The aim of this study was to test, ex vivo, if the microbiota of broilers fed a stimbiotic are better able to ferment different xylose-rich substrates in an ileal and a caecal environment. The ileal and caecal contents from broiler chickens fed a stimbiotic or from a control group were used as an inoculum in the ex vivo fermentation experiment. Different xylose-rich substrates including monomeric xylose (XYL), XOS with DP 2 to 6 (XOS), short DP XOS of 2 to 3 (sDP-XOS), long DP XOS of 4 to 6 (lDP-XOS) and de-starched wheat bran (WB), were added to each ileal and caecal inoculum in fermentation vessels. Total gas, short-chain fatty acids (SCFA) production, bacterial quantification, and carbohydrate utilisation were monitored for 9 h post-inoculation. No significant interactions were observed in any of the parameters measured in either the ileal or caecal contents (p > 0.05). Stimbiotic ileal inocula resulted in higher total gas (p < 0.001) and volatile fatty acid (VFA) (p < 0.001) production, increased numbers of Lactobacillus spp. (p < 0.001), and decreased numbers of Enterococcus spp. (p < 0.01) after 9 h regardless of the xylose-rich substrate added. Stimbiotic caecal inocula resulted in a higher ratio of VFA to branched-chain fatty acids (BCFAs) by up to +9% (p < 0.05). Ileal microbiota were found to preferentially metabolise WB, while caecal microbiota favoured XOS substrates, particularly lDP-XOS. These results indicate that stimbiotics can promote the abundance of lactic acid bacteria involved in the establishment of fibre-degrading bacteria and VFA content in the gut, which could have beneficial effects on broiler performance. Further, ileal and caecal microbiota differ in their utilisation of different substrates which may impact the effectiveness of different stimbiotic products.

Supplementation of xylo-oligosaccharides to suckling piglets promotes the growth of fiber-degrading gut bacterial populations during the lactation and nursery periods.

Modulating early-life microbial colonization through xylo-oligosacharides (XOS) supplementation represents an opportunity to accelerate the establishment of fiber-degrading microbial populations and improve intestinal health. Ninety piglets from 15 litters were orally administered once a day from d7 to d27 of lactation with either 5 mL of water (CON) or 5 mL of a solution containing 30 to 60 mg of XOS (XOS). Supplementation ceased at weaning (d28) when all piglets were fed the same commercial pre-starter diet. Growth performance did not differ between treatments during the experimental period (d7 to d40). Piglet's fecal microbiota (n = 30) shifted significantly from the end of lactation (d27) to nursery period (d40) exhibiting an increase in microbial alpha diversity. Animals supplemented with XOS showed higher richness and abundance of fiber-degrading bacteria and short-chain fatty acid (SCFA) production at d27 and d40. Additionally, the predicted abundance of the pyruvate to butanoate fermentation pathway was increased in the XOS group at d40. These results show that supplementation of XOS to lactating piglets promotes fiber-degrading bacterial populations in their hindgut. Moreover, differences observed in the nursery period suggest that XOS can influence the microbiota in the long-term.

Interaction between xylanase and a proton pump inhibitor on broiler chicken performance and gut function.

Three hundred thirty-six Ross 308 male broiler chicks were used in a 21-d study to explore performance and gut function when treated with a proton pump inhibitor (PPI; 0 or 89 mg/kg) in a 2 × 2 factorial arrangement with a xylanase (Xyl; 0 or 0.1 g/kg) to determine if the beneficial activity of arabinoxylan (AX) depolymerisation, through arabinoxylo-oligosaccharides (AXOS) production, starts in the upper gastrointestinal tract. Treatment with the PPI started from d 14, and by d 21 animal performance had deteriorated (P < 0.001). An interaction was observed between PPI and Xyl for feed conversion ratio (FCR) (P < 0.05), whereby the combination reduced the negative effect of PPI on FCR. Application of PPI raised digesta pH in the gizzard and caecum (P < 0.05), increased protein concentrations in the lower gut (P < 0.05) and reduced intake of digestible nutrients (P < 0.05). Caecal concentrations of indole, p-cresol, ammonia and the ratio of total volatile fatty acid (VFA) to butyric acid were increased with PPI (P < 0.05), indicating enhanced protein fermentation. Xylanase activity in the digesta were greatest in the caeca, especially when Xyl was supplemented (P < 0.001). The concentration of total soluble AX was greater in the gizzard and ileal digesta with Xyl supplementation (P < 0.05), supporting the depolymerisation action of xylanase even under acidic conditions. These data suggest xylanase may function in the gizzard even though pH is not optimal for activity and emphasises the importance of chlorohydric acid secretions in ensuring overall optimum gut function. AX depolymerisation benefits animal performance although it is still unknown how the AXOS produced with xylanase supplementation in the upper gastrointestinal tract influence the microbial populations and overall gut functionality.

Evaluation of xylanase and a fermentable xylo-oligosaccharide on performance and ileal digestibility of broiler chickens fed energy and amino acid deficient diets.

The objective of this study was to evaluate the effect of a product consisting of a combination of xylanase and xylo-oligosaccharide (STBIO) on performance and ileal digestibility of broiler chickens fed energy and amino acid (AA) deficient diets. Day-old male Ross 308 broiler chicks were randomly allocated to 8 pens per treatment, with 25 chicks per pen. Treatments based on wheat-corn-soybean meal diets were arranged in a 3 × 2 factorial design: a positive control that met or exceeded nutrient recommendations (PC), a negative control diet with a 50 kcal/kg apparent metabolizable energy (AME) reduction (NC1) and NC1 with a 3% reduction in AA content (NC2), each with or without supplementation of 100 g/t of the STBIO. Body weight gain (BWG), feed intake, feed conversion ratio corrected for mortality (FCR) and the European production efficiency factor (EPEF) were recorded from 0 to 42 d. On d 42, ileal samples were collected to determine dry matter (DM), organic matter (OM), ash, protein and energy digestibility. A significant interaction was observed for BWG and feed intake (P < 0.001). The energy and AA reduction reduced (P < 0.05) BWG when compared to the PC. The effect of STBIO on BWG was greater in NC1 (+451 g/bird) than in NC2 (+314 g/bird) or PC (+176 g/bird) diets (P < 0.05), and that in NC2 with STBIO was equal to that in PC without STBIO, and that in NC1 with STBIO was equal to that in PC with STBIO. No interactions were observed on the EPEF or FCR; however, STBIO improved EPEF (P < 0.001) and FCR (P < 0.001) irrespective of the energy reduction or AA density. The intake of digestible DM, OM, ash and energy for the finisher period was increased with STBIO supplementation (P < 0.01). A significant interaction was observed for the intake of digestible protein. NC1 and NC2 reduced the intake of digestible protein; however, when STBIO was supplemented, it was improved in both diets to similar levels to the PC. The stimbiotic supplementation improved performance of broiler chickens fed all diets, particularly those deficient in AME and AA.

Dietary xylanase and live yeast supplementation influence intestinal bacterial populations and growth performance of piglets fed a sorghum-based diet.

This study was to evaluate the effect of xylanase supplementation and the addition of live yeast, Saccharomyces cerevisiae, on growth performance and intestinal microbiota in piglets. One hundred and eighty commercial crossbred 23-d-old piglets (PIC 417) were sorted by initial BW and allocated to 3 treatments: control (CTR) diet, CTR diet supplemented with xylanase at 16,000 birch xylan units/kg (XYL) and XYL diet supplemented with live yeast (2 × 1010 CFU/g) at 1 kg/t (XYL + LY). Each treatment had 10 replicates, with 6 animals each. A sorghum-based diet and water were available ad libitum for 42 d of the study. Average daily gain (ADG) and average daily feed intake (ADFI) were measured from 0 to 42 d (23- to 65-d-old) and feed conversion ratio (FCR) calculated. At the end of the study, bacterial identification through 16S rRNA (V3 to V4) sequencing of the ileal and caecal digesta from one piglet per replicate was performed. No treatment effects were observed on ADFI. Pigs offered the live yeast in addition to the xylanase had increased ADG compared with those supplemented with xylanase alone (XYL + LY vs. XYL; P = 0.655). FCR was improved with XYL and XYL + LY compared with CTR (P = 0.018). Clostridiaceae counts in the ileum tended to reduce by 10% with XYL and 14% with XYL + LY compared to CTR (P = 0.07). XYL and XYL + LY increased the counts of Lactobacillaceae in the caecum compared with CTR (P < 0.0001). Dietary supplementation of live yeast combined with xylanase improved growth performance and microbial balance of piglets during the nursery phase.

Stimbiotic supplementation improved performance and reduced inflammatory response via stimulating fiber fermenting microbiome in weaner pigs housed in a poor sanitary environment and fed an antibiotic-free low zinc oxide diet.

This study investigated whether the inclusion of a stimbiotic (STB) can improve performance, influence intestinal microbiota and fermentation activity, and reduce pro-inflammatory cytokines in piglets fed a low zinc oxide diet without antimicrobial growth promotors compared to fructo-oligosaccharide (FOS) and mannan-oligosaccharide (MOS) when housed either in good sanitary (GS) or poor sanitary (PS) environments. One hundred forty-four male pigs (28-day-old) were sorted by initial body weight (BW) and allocated to one of six experimental treatments: 1) GS environment without any additive (GS-CTR); 2) GS environment with 0.01% stimbiotic (GS-STB); 3) PS environment (without cleaning and disinfection of a previously populated room) without any additive (PS-CTR); 4) PS environment with 0.01% STB (PS-STB); 5) PS environment with 0.1% MOS (PS-MOS); and 6) PS environment with 0.2% FOS (PS-FOS). Each treatment had six replicates, with four animals each. Three feeding phases, based on corn, wheat, and soybean meal were available ad libitum for the 42-days of the study. Housing piglets under PS conditions negatively influenced performance, increased plasma tumor necrosis factor alpha (TNF-α), affected the fecal microbial populations and increased concentrations of branched-chain fatty acids (BCFA) compared to GS. Stimbiotic improved 42-d-BW under PS conditions (P < 0.05) whereas MOS or FOS had no effect. On d35, plasma TNF-α was reduced with STB in PS (P < 0.05). The ratio between VFA:BCFA increased (P < 0.05) with STB, MOS or FOS in PS, and under GS condition, STB also increased the ratio. Stimbiotic increased the proportion of Clostridiales Family XIII Incertae Sedis and Clostridiaceae, while MOS and FOS increased Selenomonadaceae, Catabacteriaceae and Fibrobacteraceae. These results indicate that STB shifted the intestinal microbiome to favor fiber fermentation which likely contributed to reduced inflammatory response and improved performance, particularly in piglets reared in PS conditions.

Influence of Particle Size and Xylanase in Corn-Soybean Pelleted Diets on Performance, Nutrient Utilization, Microbiota and Short-Chain Fatty Acid Production in Young Broilers.

The objective of this study was to investigate the effects of particle size and xylanase supplementation in corn-based pellet diets on the performance and digestive traits in young broilers. A total of 512 male Ross 308 broilers were used in a 21-day study. The treatments were designed in a 4 × 2 factorial arrangement with four levels of geometric mean diameter (Dgw) of corn (540, 660, 1390, and 1700 µm), and two levels of xylanase (0 or 16,000 BXU/kg diet). Feeding coarse corn diets (1390 and 1700 µm Dgw) and xylanase supplementation showed an inferior coefficient of variation of body weight. Higher gizzard weight, microbiome alpha-diversity, and clustered separately beta-diversity (p < 0.05) were observed in birds fed coarse diets. The addition of xylanase promoted changes in relative bacteria abundance, increasing Lachnospiraceae, Defluviitaleaceae, Bacteroidaceae, Bacillaceae, Eggerthellaceae, and Streptococcaceae families in the 1700 µm group, and Christensenellaceae and Lachnospiraceae families in the 540 µm Dgw group. In conclusion, xylanase supplementation and particle size of corn interact in the intestinal environment, showing changes in microbial composition. Coarse diets and xylanase supplementation showed improved body weight homogeneity, which might be related to a better gut development and microbiota modulation.

Effect of dietary supplementation of xylanase in a wheat-based diet containing canola meal on growth performance, nutrient digestibility, organ weight, and short-chain fatty acid concentration in digesta when fed to weaned pigs.

This study was designed to determine the effect of dietary supplementation of xylanase on growth performance, nutrient digestibility, organ weight, digesta pH, and concentration of short-chain fatty acids (SCFA) of weaned pigs fed wheat-canola meal (CM) diets over a 35-d period. A total of 144 piglets (72 barrows and 72 gilts) weaned at 18 ± 2 d of age, with initial body weight (BW) of 6.2 ± 0.7 kg, received one of eight dietary treatments based on randomized complete block design. BW and feed intake were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F). Treatments consisted of a control wheat-soybean meal-based diet and wheat-regular (RCM), dehulled (DCM), or coarse CM (CCM) without and with 16,000 BXU/kg xylanase (Econase XT). All diets contained 500 FTU/kg of phytase (Quantum Blue 5G) and titanium dioxide (0.3%). Apparent total tract digestibility (ATTD) of neutral detergent fiber (NDF), crude protein (CP), phosphorous (P), calcium (Ca), dry matter, and diet were determined. On day 35, one pig per pen was euthanized to evaluate the main factors of protein, xylanase supplementation, and sex on organ weight, ileal and colon digesta pH, and colon digesta concentrations of SCFA. The main factors did not affect growth performance. Xylanase supplementation improved nutrient digestibilities in all diets and increased ileal and colonic digesta pH without affecting the growth performance of weaned pigs fed wheat and CM-based diets. A protein-xylanase effect (P < 0.05) resulted in increasing the ATTD of NDF from 28% to 32% and from 29% to 37% for RCM and DCM, respectively. The ATTD of CP was greater (P < 0.05) with xylanase supplementation (75% vs. 70%). Xylanase supplementation increased ATTD of P and Ca. A three-way interaction (P < 0.05) for protein-xylanase-gender for colon pH, acetic, and propionic acid in the colon digesta of pigs indicated that, in addition to the protein source, piglet sex could have influenced how xylanase works. Xylanase supplementation increased (P < 0.05) the weight of the liver and spleen and tended (P < 0.10) to increase the size of the kidney. In conclusion, dietary supplementation of xylanase increased nutrient digestibility and digesta pH but did not influence the growth performance of weaned pigs fed wheat and CM-based diets over a 35-d period.

The use of starter cultures in the table olive fermentation can modulate the antiadhesive properties of brine exopolysaccharides against enterotoxigenic Escherichia coli.

The present study aimed to evaluate different mates of Candida boidinii and Lactobacillus pentosus strains as starters in green table olive fermentation. Changes in fermentation characteristics as well as changes in the functional properties of the microbial exopolysaccharides (EPS) produced during the process were registered. The in vitro adhesion test demonstrated that most EPS samples could specifically attach ETEC K88. In vitro studies with porcine intestinal cells showed the improved blocking activity of the fimbria (blocking test) when the mutant strain L. pentosus 119-14MT was used alone as a starter. All EPS samples showed the ability to block receptors in the cells (exclusion test) although without differences between starter treatments. In the displacement test, EPS samples failed to remove the pathogen once attached. According to these results, L. pentosus 119-14MT, a high EPS variant, seemed to be the most effective starter improving the anti-adhesive properties of brine EPS and increasing its ability to block the ETEC K88 fimbria. These results illustrate that the anti-adhesive properties of the EPSs produced during the traditional fermentation of olives could be modulated by the use of defined starters. This opens the door to new fermentation processes aimed to produce green table olives as functional food to prevent ETEC diarrhea.

Exopolysaccharides from olive brines could reduce the adhesion of ETEC K88 to intestinal epithelial cells.

This study aims to explore the biological functions of the isolated exopolysaccharides (EPSs) produced during the industrial fermentation of olives against enterotoxigenic E. coli (ETEC) K88. Exopolysaccharides were isolated from five industrial fermenters. Analysis of their monosaccharide composition by GLC revealed that the main components were glucose (27%-50%) and galactose (23%-33%) followed by rhamnose (4-23%) and arabinose (6-17%). The 1H NMR spectrum showed a very similar profile between samples, and a more in-depth analysis revealed the presence of an α-pyranose in the form of α-d-Glcp-(1→) and two different α-furanoses, with chemicals shift values, suggesting the presence of α-d-Glcf and α-d-Galf. Miniaturized in vitro tests demonstrated the ability of EPS samples to attach specifically to ETEC K88 (P < 0.05) with variable intensities. The competition test did not show the ability to block the ETEC K88 adhesion to IPEC-J2 cells; however, in the displacement test, all EPS samples were shown to effectively remove the pathogens attached to the cells (P < 0.01). These results suggest that the EPSs produced during the fermentation of table green olives could interfere with the attachment of opportunistic pathogens onto the intestinal epithelial cells. This would open the possibility of novel functional properties for this traditional Mediterranean fermented food and for the isolated EPSs as candidates for nutraceutics to be used in human and/or animal diets in the prevention and treatment of ETEC diarrhoea.

Xylanase, protease and superdosing phytase interactions in broiler performance, carcass yield and digesta transit time.

The interaction of xylanase, protease and superdosing (1,500 FTU/kg) phytase in a 2 × 2 × 2 factorial arrangement was studied in broilers fed sorghum-based diets. A total of 2,800 one-day-old unsexed Ross 308 chicks were housed in 56 pens with 50 birds per pen, with or without inclusion of xylanase, protease and phytase, totaling 8 treatments and 7 replicates per treatment. Body weight (BW) and feed intake (FI) were measured at 21 and 42 days of age, and mortality corrected feed conversion ratio (FCR) was calculated for each period and cumulatively. Tibia ash and carcass yield were determined in 2 birds per replicate at 21 and 42 days of age, respectively. Digesta transit time was determined at 21, 28, 35 and 42 days of age using 5 birds per replicate. Results showed that superdosing phytase increased BW and FI at 42 days of age (P < 0.05) and xylanase improved FCR (P < 0.05). Xylanase and phytase also positively influenced carcass yield and breast weight, respectively. Overall, inclusion of superdosing phytase increased transit time when included in a diet containing xylanase, and no change with protease inclusion. In conclusion, the beneficial effects of xylanase, protease and superdosing phytase in broiler performance were not additive. This limitation is likely not related to the lack of efficacy of any one of the individual enzymes but to a limitation of the bird to respond additively to successive additions of enzymes.

Evaluation of the effect of different wheats and xylanase supplementation on performance, nutrient and energy utilisation in broiler chicks.

The aim of this study was to evaluate the performance, nutrient utilisation and energy metabolism of broiler chicks fed 8 different wheat samples, supplemented or not with xylanase. Seven-hundred sixty eight male broilers (1-day-old) were distributed to 16 experimental treatments (6 replicates per treatment). The treatments were in a factorial arrangement with 8 different wheats and 2 levels of xylanase (0 or 16,000 BXU/kg). The predicted apparent metabolisable energy (AME) of the wheat samples ranged from 13.0 to 13.9 MJ/kg and all diets were formulated to contain the same amount of wheat. Body weight gain (BWG) and feed intake (FI) were measured at 21 d, as was jejunal digesta viscosity, and feed conversion ratio (FCR) calculated. On day 24, one representative bird per pen was selected to calculate whole body energetics. At 21 d, 3 chicks per replicate were randomly allocated to metabolism cages for energy and nutrient utilisation determinations, and were continued on the experimental diets until 24-d-old. No interactions were observed for any performance response variables, ileal nutrient utilisation or digesta viscosity. Xylanase improved BWG and reduced FCR and digesta viscosity (P < 0.05). Wheat influenced dry matter (DM) utilisation and xylanase increased ileal digestible energy (P = 0.04). Xylanase also improved (P < 0.05) DM and nitrogen retention. Apparent metabolisable energy and AME corrected for nitrogen (AMEn) were subject to an interaction whereby wheats 2 and 6, which returned the lowest AME and AMEn values, responded to xylanase supplementation and the remainder did not. Net energy for production and the efficiency of energy use for production were not influenced by xylanase, but were affected by wheat (P < 0.05). Despite the significant differences between wheats with regards to their nutrient utilisation and energy metabolism in birds, xylanase removed this variance and resulted in more homogeneous performance.

A proteinaceous fraction of wheat bran may interfere in the attachment of enterotoxigenic E. coli K88 (F4+) to porcine epithelial cells.

Wheat bran (WB) from Triticum aestivum has many beneficial effects on human health. To the best of our knowledge, very little has been published about its ability to prevent pathogenic bacterial adhesion in the intestine. Here, a WB extract was fractionated using different strategies, and the obtained fractions were tested in different in vitro methodologies to evaluate their interference in the attachment of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal porcine epithelial cells (IPEC-J2) with the aim of identifying the putative anti-adhesive molecules. It was found that a proteinaceous compound in the >300-kDa fraction mediates the recognition of ETEC K88 to IPEC-J2. Further fractionation of the >300-kDa sample by size-exclusion chromatography showed several proteins below 90 kDa, suggesting that the target protein belongs to a high-molecular-weight (MW) multi-component protein complex. The identification of some relevant excised bands was performed by mass spectrometry (MS) and mostly revealed the presence of various protease inhibitors (PIs) of low MW: Serpin-Z2B, Class II chitinase, endogenous alpha-amylase/subtilisin inhibitor and alpha-amylase/trypsin inhibitor CM3. Furthermore, an incubation of the WB extract with ETEC K88 allowed for the identification of a 7S storage protein globulin of wheat, Globulin 3 of 66 kDa, which may be one of the most firmly attached WB proteins to ETEC K88 cells. Further studies should be performed to gain an understanding of the molecular recognition of the blocking process that takes place. All gathered information can eventually pave the way for the development of novel anti-adhesion therapeutic agents to prevent bacterial pathogenesis.

New properties of wheat bran: anti-biofilm activity and interference with bacteria quorum-sensing systems.

Some plant extracts, have been demonstrated to interfere with the microbial metabolism of several pathogenic bacteria. Within this antimicrobial properties it has been described the potential to inhibit or destroy biofilms or to interfere in quorum-sensing (QS) systems. However, to our knowledge, no study exploring this potential of wheat-bran (WB) has been published. The purpose of the present study is to evaluate the anti-biofilm activity of WB against a cow mastitis strain of Staphylococcus aureus and also its possible interference with bacterial QS systems. The potential of inhibition and destruction of the biofilm was studied by different in vitro assays. Also, we tested the ability of WB to interfere in bacterial QS by degrading acyl-homoserine lactones (AHL) as one of the most studied QS signal molecules for Gram-negative bacteria. The soluble extract of WB at 0.5% showed anti-biofilm activity, inhibiting biofilm formation and also destroying it. Similarly, the > 300 kDa fraction from WB had significant anti-biofilm activity in both in vitro assays. The WB also showed a potential to interfere with bacterial QS systems, as it was demonstrated to contain certain lactonase activity able to reduce AHL concentration in the medium. The present study reveals two additional beneficial properties of WB extract never explored before, which may be related to the presence of defence compounds in the plant extract able to interfere with microbial biofilms and also QS systems.

Screening the ability of natural feed ingredients to interfere with the adherence of enterotoxigenic Escherichia coli (ETEC) K88 to the porcine intestinal mucus.

The inhibition of the attachment of bacteria to the intestine by receptor analogues could be a novel approach to prevent enterotoxigenic Escherichia coli (ETEC) K88-induced diarrhoea in piglets. The objective of the present study was to screen the ability of different feed ingredients (FI) to bind to ETEC K88 (adhesion test, AT) and to block its attachment to the porcine intestinal mucus (blocking test, BT) using in vitro microtitration-based models. In the AT, wheat bran (WB), casein glycomacropeptide (CGMP) and exopolysaccharides exhibited the highest adhesion to ETEC K88 (P< 0·001). In the BT, WB, CGMP and locust bean (LB) reduced the number of ETEC K88 attached to the intestinal mucus (P< 0·001). For WB and LB, fractionation based on their carbohydrate components was subsequently carried out, and each fraction was evaluated individually. None of the WB fractions reduced the adhesion of ETEC K88 to the mucus as did the original extract, suggesting that a protein or glycoprotein could be involved in the recognition process. With regard to the LB fractions, the water-extractable material reduced the adhesion of ETEC K88 (P< 0·001) to the mucus similar to the original extract (P< 0·001), indicating, in this case, that galactomannans or phenolic compounds could be responsible for the recognition process. In conclusion, among the FI screened, the soluble extracts obtained from WB, LB and CGMP exhibited the highest anti-adhesive properties against ETEC K88 in the BT. These results suggest that they may be good candidates to be included in diets of weaned piglets for the prevention of ETEC K88-induced diarrhoea.

Effects of dietary supplementation of Bacillus amyloliquefaciens CECT 5940 and Enterococcus faecium CECT 4515 in adult healthy dogs.

The aim of this study was to evaluate the effect of a probiotic preparation consisting of two probiotic strains, Bacillus amyloliquefaciens CECT 5940 and Enterococcus faecium CECT 4515 (each 5 · 10(8) CFU/g feed), on faecal consistency, faecal microbiology and nutrient digestibility in adult healthy dogs. Sixteen beagles (eight males and eight females) were divided into two groups: the Control group (CON), which was fed the basal diet, and the probiotic group (PRO), which received the basal diet supplemented daily with 1 · 10(8) CFU for 39 consecutive days. Faecal score was assessed before (BS) and throughout the supplementation period (SP). Fresh faecal samples were collected before supplementation, before finishing the supplementation period and after 6 days of withdrawal for microbial enumeration and pH measurement. During the supplementation period, a digestibility trial was performed. There were no differences in faecal scores or the digestibility coefficients between groups. Between groups no statistical differences were found in most microbiota analysed or in faecal pH. However, during the supplementation period, pathogenic clostridia dropped significantly in Group PRO (5.64 vs. 2.94 ± 0.53 CFU/g faeces; p < 0.001), when compared with the period BS. The use of the probiotic preparation had no impact on nutrient digestibility by adult healthy dogs; however, it could stabilise faecal microbiota by decreasing pathogenic clostridia.

Consumption of dietary n-3 fatty acids decreases fat deposition and adipocyte size, but increases oxidative susceptibility in broiler chickens.

The aim of this study was to determine the effects of an omega-3 (n-3) polyunsaturated fatty acid (PUFA)-enriched diet on animal fat depots and lipid oxidation in the blood and meat of broiler chickens. Abdominal fat pad (AFP), sartorius muscle and liver histology were used to assess the effect of the dietary fat on animal lipid depots. A total of 60 female broilers (14 days old) was randomly divided into two groups which received a diet containing 10 % of tallow (S diet), rich in saturated fatty acids or 10 % of a blend of fish oil and linseed oil (N3 diet), rich in n-3 PUFA from 14 to 50 days of life. Both absolute and relative weights of AFP in N3 animals were lower than in the S group (P < 0.05). These results paralleled with a lower adipocyte mean area (P < 0.001) obtained in N3-fed animals, leading to a higher number of fat cells per unit of surface measured (383.4 adipocytes/mm(2) vs. 273.7 adipocytes/mm(2)). Similarly, fat content and the intramuscular fat-occupied area of muscle were lower in N3 (P < 0.0001) than in the S-fed birds. Neither macroscopic nor microscopic differences were observed in the liver. The inclusion of dietary n-3 PUFA increased meat and erythrocyte oxidation susceptibility; however, the erythrocytes from the S group were less resistant to osmotic changes. Results indicate that feeding an n-3 PUFA diet influences fat distribution and the oxidative status of broiler chickens.