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SUMMARY Direct-Fed Microorganisms (DFM) and exogenous enzymes have been demonstrated to improve growth performance in poultry and are potentially important components of management programs for antibiotic-free (ABF) poultry production. In this study, we evaluated the effect of a feed additive composed of a DFM product containing spores of 3 Bacillus amyloliquefaciens strains and a multi-enzyme complex of xylanase, amylase, and protease on the performance of broilers with or without an antibiotic growth promoter (AGP), either bacitracin methylene disalicylate (BMD) or virginiamycin. Growth performance of broilers fed one of 6 treatments arranged in a 3 (AGP) × 2 (additive) factorial was evaluated over a 42 d production period. Overall, administration of the feed additive improved body weight, body weight gain, and feed conversion ratio. Administration of either antibiotic significantly reduced cumulative FCR when compared to the control, while BMD administration reduced foot pad lesions as compared to the control. Our results suggest the co-administration of DFM and enzymes may be an important component of ABF poultry production programs and the benefits of their use are not dependent on the presence or absence of AGP.

This study evaluated the effects of a combination of xylanase, amylase, and protease (XAP), with probiotics (3 Bacillus spp.) supplementation on apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in Cobb 500 broilers from 0 to 21 d. A completely randomized 2 × 4 factorial design (2 levels of fiber; 4 types of supplements) with 8 replicate cages (6 birds/cage) was used. Each low and high-fiber diet contained 500 FTU/kg Buttiauxella sp. phytase and was supplemented with: (a) none (control), (b) XAP (2,000 U xylanase + 200 U amylase + 4,000 U protease/kg diet), (c) probiotics (75,000 CFU/g of Bacillus spp.), or (d) XAP + probiotics. High fiber decreased (P < 0.05) nitrogen-corrected apparent metabolizable energy (AMEn), AID of all amino acids (AA), AID and ATTD of dry matter (DM), crude protein (CP), starch, and gross energy (GE). High fiber increased (P < 0.01) the flow of total non-starch polysaccharides (NSP) in both ileum and total tract. The XAP + probiotics increased (P < 0.01) AMEn as well as AID and ATTD of DM, CP, GE, starch, while alone, XAP yielded similar improvement except for DM compared with control. The supplemental XAP alone improved (P < 0.01) the digestibility of most of the AAs compared with control. Moreover, XAP + probiotics increased (P < 0.05) AID of all AA except arginine and serine compared with control. A fiber × supplements interaction (P < 0.05) was found for AID of histidine and threonine, and their digestibility in high-fiber diet was improved to a level comparable to low-fiber diet by XAP + probiotics. The flow of NSP in XAP group was 5 to 6% lower than in control while NSP flow in XAP + probiotic group was further 4% lower than that of XAP group (P < 0.01). The results infer that the combination of XAP and probiotics can effectively optimize the nutrient digestibility in broilers fed both low and high-fiber diets.

Direct-fed microorganisms (DFM) and exogenous enzymes have been demonstrated to improve growth performance in poultry and are potentially important alternatives to antibiotic growth promoters (AGP). We investigated the administration of a feed additive composed of a DFM product containing spores of 3 Bacillus amyloliquefaciens strains and an enzyme blend of endo-xylanase, α-amylase, and serine-protease in diets with or without sub-therapeutic antibiotics in broiler chickens over a 42-d growth period. Evaluation of growth performance determined feed efficiency of broiler chickens which were administered the feed additive was comparable to those fed a diet containing AGPs. Characterization of the gastrointestinal microbiota using culture-dependent methods determined administration of the feed additive increased counts of total Lactic Acid Bacteria (LAB) relative to a negative control and reduced Clostridium perfringens to levels similar to antibiotic administration. Additionally, greater counts of total LAB were observed to be significantly associated with reduced feed conversion ratio, whereas greater counts of C. perfringens were observed to be significantly associated with increased feed conversion ratio. Our results suggest the co-administration of DFMs and exogenous enzymes may be an important component of antibiotic free poultry production programs and LAB and C. perfringens may be important targets in the development of alternatives to AGPs in poultry production.

A 26-day trial with 18 Pietrain × (Landrace × Duroc) pigs was conducted to investigate the effect of two dose levels of a specifically selected Bacillus spp. direct-fed microbial (DFM) product, on the emission of environmentally harmful gasses (methane, ammonia and hydrogen sulphide) from manure. Pigs were assigned to one of three treatments in a randomized complete block design according to their sex and initial BW. Each treatment contained three replications with two pigs per pen. The test treatments included a Bacillus spp. DFM containing 3 × 108 colony-forming unit/g, added at a low (250 mg/kg) and high (500 mg/kg) dose to an antibiotic free high fibre-based diet, and a non-supplemented control diet. Manure from pigs fed with the supplemented diets emitted lower amounts of atmospheric contaminants. The most significant reduction was observed with low DFM supplementation, in which methane and ammonia volatilization decreased ( P< 0.05) by >40% and 50%, respectively, on fresh weight basis in relation to the control. Microbiome analysis of manure by high through put sequencing techniques on eubacterial and archaeal 16S rRNA genes highlighted the complex interactions between indigenous gut microflora and inoculated Bacillus spp. The tested Bacillus DFM could be considered as a best available technique in reducing the environmental impacts of growing pigs fed with high fibre-based diets. info:eu-repo/semantics/publishedVersion

This study evaluated the effect of a direct-fed microbial (DFM) and its combination with xylanase, amylase, and protease (XAP) enzymes cocktail on performance of broilers, in comparison with two commonly used antimicrobial growth promoters (AGPs). Five treatments were tested using day-old Ross 708 broilers (mixed sex at 50:50 ratio) in a completely randomized design with 8 replications per treatment (40 birds/pen) using built up litter. Treatments were 1) a negative control (NC) based on corn/soy/wheat (10%) containing 500FTU/kg phytase, 2) NC + DFM (150,000 CFU/g of 3 Bacillus strains) (DFM150); 3) NC+ DFM (75,000 CFU/g of 3 Bacillus strains) in combination with XAP, XAP + DFM75); 4) NC+ BMD (50 g/ton), and 5) NC+ virginiamycin (20 g/ton). Diets were fed in mash form ad libitum in 3 phases: d 1 to 10 (starter); d 11 to 21 (grower) and d 22 to 42 (finisher). Body weight and feed intake (FI) were measured per phase and feed conversion ratio (FCR) was corrected for mortality weight. Treatment means were compared using a one-way analysis of variance (ANOVA) and separated via Duncan's Multiple Range Test. Supplementation of DFM increased (P

Improving the efficiency of P use by pigs is especially important for lactating sows, whose metabolic requirements for P and Ca are high. The effect of a sp. phytase on lactating sow performance and nutrient digestibility was investigated using the combined data set for 6 studies. Treatments included a nutritionally adequate positive control diet (PC), a negative control diet (NC; with an average reduction of 0.16% available phosphorous and 0.15% Ca vs. PC), and NC supplemented with a sp. phytase at 250, 500, 1,000 or 2,000 phytase unit (FTU)/kg, respectively. Phosphorus and Ca deficiency in the NC resulted in significantly higher BW loss compared with the PC. All phytase treatments maintained BW loss at the same level as the PC. Increasing doses of phytase significantly ( < 0.05) reduced sow BW loss and increased energy intake, with improvements most apparent in sows older than parity 5. The positive effects on BW and energy intake were not observed in first-parity sows. This may be a consequence of fewer first parity sows in the data set. The apparent total tract digestibility of DM, OM, and CP were not affected by phytase supplementation. Digestible P and Ca were significantly improved (linear, < 0.0001; quadratic, < 0.0001) by increasing the dose of phytase supplementation. Significantly lower apparent total tract digestibility of energy, Ca, and P was found in the NC treatment vs. the PC treatment, whereas no significant differences were found between phytase treatment and the PC treatment. In conclusion, phytase supplementation at a level of 250 FTU/kg can replace 0.16% available phosphorous and 0.15% Ca; however, increasing the phytase dose can further reduce BW loss in sows fed P- and Ca- deficient diets.

This study evaluated the effects of nutritional supplementation with a blend of essential oils (EO, thymol and cinnamaldehyde) and an enzyme combination (XB, xylanase and β-glucanase), given alone or together, on growth performance, microbial counts, ileum microscopic anatomy and the expression of inflammatory mediators in weaned piglets challenged with Escherichia coli K88. One hundred and ninety-two weaned piglets were allocated to 8 treatments in a 35-day experiment with a 2 × 4 factorial design that compared 2 levels of oral E. coli challenge [sham (−) or infected (+)] under different dietary treatments [fed the basal diet (CTR) either with or without EO or XB individually or in combination (EOXB)]. Half of the piglets were orally challenged with E. coli O149:F4 (K88) on day 8 and 48 piglets (1 piglet/pen) were slaughtered on day 35. The E. coli challenge was found to decrease the average daily gain (ADG) and the gain to feed ( G : F ) ratio from days 7 to 14 ( P P P = 0.02). The E. coli challenge increased the populations of fecal Clostridia , E. coli and coliforms on day 9 ( P E. coli and coliforms counts on day 14 ( P P = 0.01, respectively). Dietary EO and EOXB reduced the fecal coliforms count compared to the CTR group on day 14 ( P = 0.02 and P P E. coli count ( P = 0.051) compared to the CTR group in the cecum digesta. Dietary EOXB also decreased the crypt depth and increased the villus-to-crypt ratio in piglets compared to the CTR group ( P E. coli challenge up-regulated the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1α, toll-like receptor (TLR)2 and TLR4 ( P P = 0.01, P = 0.03, P = 0.01, respectively), and EOXB supplementation down-regulated the expression of TNF-α ( P P = 0.046) compared to the EO group. The results suggest that the combination of EO and XB supplementation may have beneficial effects on the modulation of fecal consistency, microbial counts and ileum microscopic anatomy in weaned piglets following E. coli infection.

Primary Audience: Nutritionist, Poultry feed producer, Poultry farmers SUMMARY The objective of this study was to determine the response of broilers to the combination of multi-enzymes and direct-fed microbial (DFM) under commercial production settings. A total of 7,000 1-day-old male broilers (Ross 308) were distributed over 10 pens (700 broilers/pen). Two dietary treatments were tested using complete randomized design, including a control diet and a test diet with addition of multi-enzymes (xylanase, amylase, and protease (XAP)] and DFM (a combination of spores from 3 strains of Bacillus amyloliquefaciens). Pelleted diets were offered ad libitum in 3 phases and water was freely available. During starter and grower phases (0 to 21 d), the enzyme and DFM combination resulted in improved FE (P < 0.05). During the finisher phase, higher feed intake and BW gain (P

SummaryThe effect of a Buttiauxella phytase, in a dose range of 0-1000 FTU/kg in feed, on production performance in growing/finishing pigs fed European type wheat, corn, barley and SBM based diet was determined. Five dietary treatments were tested including a positive control (PC), a negative control (NC) without inorganic P and with reduced Ca (–0.12%) and metabolisable energy content (–0.14 MJ ME/kg) or NC supplemented with Buttiauxella phytase (Axtra® PHY, Danisco Animal Nutrition, DuPont Industrial Biosciences, Marlborough, UK) at 250, 500 or 1000 FTU/kg respectively. A total of 100 pigs (mean initial BW of 30 kg, Topig x Pi, 10 weeks of age) were used, with 20 replicates per treatment (50% gilts and 50% barrows). Pigs were individually housed in pens and randomly allocated on the basis of body weight (BW) and gender to the five treatments. Feed, in mash form, and water were offered ad libitum. Basal diets had an intrinsic phytase level of approximately 300 and 440 FTU/kg in NC grower and NC finisher diets respectively. Mean average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were calculated for the feeding phases of 30–85 kg (grower), 85–125 kg (finisher) on trial and for overall period. During the grower phase, increasing phytase dose resulted in a linear increase in ADG. Phytase inclusion at 250, 500 and 1000 FTU/kg improved ADG by 3.5, 7.2 and 8.1% respectively compared to NC and by 0.8, 4.5 and 5.3% respectively compared to those fed the PC. Performance in the finisher phase, overall period and slaughter parameters were not different between dietary treatments. The data showed that it was beneficial to increase phytase dose up to 1000 FTU/kg in grower pigs up to 85 kg BW. In the finisher phase, in pigs with BW above 85 kg, 500 FTU/kg was sufficient to replace inorganic P and maintain performance and carcass characteristics comparable to pigs fed the PC.

This review focuses on phytase functionality in the digestive tract of farmed non-ruminant animals and the factors influencing in vivo phytase enzyme activity. In pigs, feed phytase is mainly active in the stomach and upper part of the small intestine, and added phytase activity is not recovered in the ileum. In poultry, feed phytase activities are mainly found in the upper part of the digestive tract, including the crop, proventriculus and gizzard. For fish with a stomach, phytase activities are mainly in the stomach. Many factors can influence the efficiency of feed phytase in the gastrointestinal tract, and they can be divided into three main groups: (i) phytase related; (ii) dietary related and (iii) animal related. Phytase-related factors include type of phytase (e.g. 3- or 6-phytase; bacterial or fungal phytase origin), the pH optimum and the resistance of phytase to endogenous protease. Dietary-related factors are mainly associated with dietary phytate content, feed ingredient composition and feed processing, and total P, Ca and Na content. Animal-related factors include species, gender and age of animals. To eliminate the antinutritional effects of phytate (IP6), it needs to be hydrolyzed as quickly as possible by phytase in the upper part of the digestive tract. A phytase that works over a wide range of pH values and is active in the stomach and upper intestine (along with several other characteristics and in addition to being refractory to endogenous enzymes) would be ideal. © 2014 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Two trials (a 42-d performance and a 21-d cohort digestibility) were conducted to evaluate the performance and nutrient digestibility of broilers fed corn diets supplemented with exogenous xylanase, amylase, and protease as single or combined activities. A nutritionally adequate, positive control (PC) diet was formulated. The negative control (NC) diet was formulated to be lower in metabolizable energy (∼86 kcal/kg diet) and digestible amino acids (1 to 2%) compared to PC. The other 4 treatments were based on the NC and they were either supplemented with xylanase (X), amylase (A), protease (P), or a combination of X, A, and P (XAP; to provide 2,000 U of X, 200 U of A, and 4,000 U of P/kg diet). All diets were marginal in AvP and Ca and contained a background of phytase (1,000 FTU/kg). In each trial, male broiler (Ross 308) chicks were allocated to the 5 treatments (10 replicates of 20 birds/pen and 9 replicates of 8 birds/cage for the performance and digestibility trials, respectively). In the digestibility trial, ileal digesta was collected on d21 for the determination of nutrient utilization. Data were subjected to one-way ANOVA and means were separated by Tukey's HSD test. Only the XAP improved (P 0.05) by dietary treatments. Both X and XAP had lower (P 0.05) FCR compared to PC. In conclusion, these results suggest a synergistic effect between X, A and P on broiler performance and nutrient digestibility. In the current study, AIDE measurements appeared to overestimate the enzyme response. Calculation of the energy contribution by supplemental enzymes using the improvements in the digestibility of the undigested fraction of starch, fat and protein may be a more accurate measurement for the enzyme response than the absolute response in AIDE.

SummaryThis study evaluated the effect of an enzyme blend (xylanase, amylase and protease; XAP) in combination with a direct fed microbial (DFM) containing three strains of Bacillus spp. on intestinal histology, immune response and performance of broilers. Four dietary treatments were tested in a 2 × 2 factorial trial, including two levels of challenge (without or with coccidial infection), two levels of feed additive (with or without XAP and DFM). Diets were fed ad libitum to male Cobb500 broilers in mash feeds from 1–21 days of age, with eight replicate pens per treatment within brooder-batteries with raised wire floors and built up litter, housing six birds per pen. A mild challenge was introduced by oral gavage at day five to the challenged birds, using a six-fold concentration of coccidial vaccine. A high fibre basal diet formulated with rye and wheat middlings was used to further increase the challenge. Body weight and feed intake were measured and feed conversion ratio (FCR) was calculated during starter (1–12 d), grower (12–21 d) and overall 1–21 days. Intestinal morphology and immune response parameters were measured on day 12 and 21. Compared to the unchallenged groups, the coccidial challenge reduced (P < 0.05) body weight gain (BWG), increased FCR, reduced villus height and increased crypt depth. The challenged birds had increased pro-inflammatory cytokines (IL-6, IL-1β; P < 0.05) in the intestine as well as higher levels of acute phase proteins (APP, haemopexin and α−1-acid glycoprotein) in the plasma and circulating heterophils. XAP + DFM supplementation improved BWG, reduced FCR and increased energy efficiency compared to the non-supplemented groups. The combination of XAP and DFM reduced inflammatory responses such as APP compared to the challenged control group and maintained performance to a comparable level seen in the unchallenged control. The data indicate that XAP enzymes in combination with Bacillus-based DFM may reduce the damage and performance losses induced by coccidial challenge.

In pig production, within-batch variation in body weight (BW) gain of piglets during the nursery period (up to 10 weeks of age) can be high and is of high economic importance. Homogeneity of BW within batches of animals is important as it influences the efficiency of use of the grower and finisher facilities, and provides an extra value for the fattening farms. In the current study, factors for a low BW at the end of the nursery period of pigs were determined by analysing datasets from three different Swine Research Centres in the Netherlands and France. The entire dataset contained information on 77,868 individual piglets born in the period between 2005 and 2010. BW was determined at different time points over the pre- and post-weaning phase, and sex, season of birth; litter information (litter size at day of birth and after cross-fostering, number of piglets born alive per litter, number of total born littermates, sow parity number); cross-fostered animals (yes or no), and pen group size over the post-weaning period were recorded. A risk factor analysis approach was used to analyze the datasets to determine factors that predict piglet BW at the end of the nursery period. BW at the end of the nursery period corrected for age was mainly determined by season (P 0.05) contribute to explaining the BW at the end of the nursery period. To discard the possibility of intrauterine growth retarded piglets (IUGR) being the reason for the influence of birth weight (BiW) as an explanatory factor in the regression model, a further analysis was performed on the effect of this category of piglets on the results of the regression analysis. Overall, it was concluded that piglet's BW at the end of the nursery phase is mainly determined by season, sex, birth and weaning weight and BW at 6 weeks of age. Piglets with a BiW higher than the mean BiW minus 2.5 times the SD have the potential to compensate during the subsequent phases of growth.

The apparent digestibility of energy-yielding nutrients (carbohydrate, protein, and fat) was predicted in the human upper digestive tract and large bowel separately for 4 diverse diets containing either a single dietary fiber source [wheat bran and pectin (PE) diets] or mixed fiber sources [low-fiber (LF) and high-fiber (HF) diets). A human balance study was undertaken to determine fecal energy and nutrient excretion and a rat model was used to predict human ileal energy and nutrient excretion. Total tract energy digestibility ranged from 92 (HF diet) to 96% (PE diet and LF diet), while at the ileal level it ranged from 79 to 86% for the HF diet to the LF diet. The predicted upper-tract digestion of starch, sugars, and fat was high, with ileal digestibilities exceeding 90% for all diets. Nonstarch polysaccharides were poorly digested in the upper tract for all diets except in the PE diet. The daily quantity of protein excreted at the ileal level was between 2 (HF diet) and 5 (PE diet) times higher than that at the fecal level. The large differences between fecal and ileal nutrient loss highlight that fecal digestibility data alone provide incomplete information on nutrient loss. There is a need to be able to routinely determine the uptake of energy in the upper and lower digestive tracts separately.

BACKGROUND:Endogenouslysineinilealdigestahasbeendetermineddirectlyusinghighlydigestibleproteinsandhydrolysates and assuming a complete absorption of dietary amino acids. In this study, the endogenous lysine in the ileal digesta of rats fed casein was determined using the guanidination method (baseline) and directly (assuming complete digestion and absorption) for casein and a highly hydrolysed casein. The peptide alimentation/ultrafiltration technique was also used to determine the endogenous ileal lysine for the casein hydrolysate (Peptopro). RESULTS: Mean endogenous ileal lysine for rats given the unguanidinated casein and determined directly assuming complete digestion and absorption (644 mg kg −1 dry matter intake (DMI)) or for a highly hydrolysed casein with the same assumption (596 mg kg −1 DMI) was significantly (P < 0.05) higher than the endogenous ileal lysine for casein determined using the guanidination method (274 mg kg −1 DMI). The endogenous lysine in ileal digesta for a casein hydrolysate determined using the alimentation/ultrafiltration method (302 mg kg −1 DMI) was not significantly (P < 0.05) different from that based on the guanidination method but was significantly (P < 0.05) lower than the equivalent endogenous lysine determineddirectly. CONCLUSION: Dietary amino acids are not completely absorbed from casein or its hydrolysate and determining endogenous amino acids in ileal digesta directly, assuming complete absorption, overestimates the amount of endogenous ileal amino acids. c � 2009 Society of Chemical Industry

Background: Systems to calculate metabolizable energy (ME) in foods and diets are often based on Atwater factors. The accuracy of these factors with low-fat diets high in fiber is unknown when food intake is reduced spontaneously. Objective: The objective was to evaluate the accuracy of Atwater factors and other systems for calculating ME available from low-fat, high-fiber diets when food intake was reduced spontaneously. Design: The ME contents of a high-fat, low-fiber diet and 2 low-fat diets, one high in fruit and vegetable fiber and the other high in cereal fiber, were determined in a randomized parallel study in humans (n 27) and compared with various factorial and empirical models for calculating ME. Results: Food intakes decreased with both the high fruit and vegetable fiber and cereal fiber diets. The difference between ME calculated by using Atwater and similar factors and determined ME values was up to 4% for the refined diet and up to 11% for the low-fat, high-fiber diets. Various factorial and empirical systems for calculating food energy failed to reflect the results of the direct determinations. Conclusion: Atwater factors were inaccurate with low-fat, highfiber diets. Although modified Atwater factors may be accurate under standardized conditions of zero-nitrogen and zero-energy balance, they overestimate energy availability from high-fiber fruit and vegetable and cereal diets when food intake is reduced spontaneously in addition to when intake is reduced voluntarily. Am J Clin Nutr 2007;86:1649 –56.

There is increasing evidence showing that dietary supplementation with prebiotics can be effective in the treatment of intestinal inflammation. Because weaning time is characterized by rapid intestinal inflammation, this study investigated the effect of a diet supplemented with a combination of 4 fermentable carbohydrates (lactulose, inulin, sugarbeet pulp, and wheat starch) on the mRNA content of proinflammatory cytokines in newly weaned piglets. Cytokines (IL-1beta, IL-6, IL-8, IL-12p40, IL-18, and tumor necrosis factor-alpha) were analyzed using a semiquantitative reverse-transcription PCR technique on d 1, 4, and 10 in the ileum and colon of piglets fed either a test diet (CHO) or a control diet. In addition to the diet, the effect of enforced fasting on cytokine mRNA content was also evaluated. No effect of fasting was observed on the pro-inflammatory cytokine mRNA content. Our results showed that the CHO diet induced an up-regulation of IL-6 mRNA content in the colon of piglets 4 d postweaning. This up-regulation was specific for the animals fed the CHO diet and was not observed in animals fed the control diet. An increase in IL-1beta mRNA content was also observed on d 4 postweaning in all of the piglets. Correlations between proinflammatory cytokines and the end-products of fermentation indicated that the regulation of cytokines may be linked with some of the fermentation end-products such as branched-chain fatty acids, which are in turn end-products of protein fermentation.

Reducing the CP content and increasing the fermentable carbohydrates (FC) content of the diet may counteract the negative effects of protein fermentation in newly weaned piglets fed high-CP diets. To study the synergistic effects of CP and FC on gut health and its consequences for growth performance, 272 newly weaned piglets (26 d of age, 8.7 kg of BW) were allotted to 1 of 4 dietary treatments in a 2 x 2 factorial arrangement, with low and high CP and low and high FC content as the factors. Eight piglets from each dietary treatment were killed on d 7 postweaning. Feces and digesta from ileum and colon were collected to determine nutrient digestibility, fermentation products, and microbial counts. In addition, jejunum tissues samples were collected for intestinal morphology and enzyme activity determination. During the entire 4-wk period, interactions between the dietary CP and FC contents were found for ADFI (P = 0.022), ADG (P = 0.001), and G:F (P = 0.033). The high-FC content reduced ADFI, ADG, and G:F in the low-CP diet, whereas the FC content did not affect growth performance in the high-CP diet. Lowering the CP content of the low-FC diet improved ADFI and ADG, whereas lowering the CP content of the high-FC diet did not influence growth performance. The low-CP diets resulted in a lower concentration of ammonia in the small intestine (P = 0.003), indicating reduced protein fermentation. In the small intestine, the high FC content increased the number of lactobacilli (P = 0.047), tended to decrease the number of coliforms (P = 0.063), tended to increase the lactic acid content (P = 0.080), and reduced the concentration of ammonia (P = 0.049). In the colon, the high-FC diets increased the concentration of total VFA (P = 0.009), acetic acid (P = 0.003), and butyric acid (P = 0.018), and tended to decrease the ammonia concentration (P = 0.076). Intestinal morphology and activity of brush border enzymes were not affected by the diet, although maltase activity tended to decrease with increasing dietary FC (P = 0.061). We concluded that an increase in the dietary FC content, and to a lesser extent a decrease in the CP content, reduced ammonia concentrations and altered the microflora and fermentation patterns in the gastrointestinal tract of weaned piglets. However, these effects were not necessarily reflected by an increased growth performance of the piglets.

An experiment was conducted to examine changes in VFA and ammonia concentrations at different time points using 4 fermentable carbohydrate-rich feed ingredients as substrates and feces of unweaned piglets as inoculum. Fecal inoculum was collected, pooled, and mixed from 9 specially raised (no creep feed or antibiotics) crossbred piglets at 3 wk of age. Inulin, lactulose, molasses-free sugar beet pulp, and wheat starch were used as substrates and were fermented in vitro for 72 h (3 replicates per substrate). Cumulative gas production was measured as an indicator of the kinetics of fermentation. In addition, 3 bottles of substrate per time point with similar contents (amounts of substrate, inoculum, and media) were incubated but were allowed to release their gas throughout incubation. For these latter bottles, fermentation fluid was sampled at incubation time points including every hour between 1 and 24 h and at 48 h, and fermentation end products (VFA, lactate, and ammonia) and OM disappearance were measured. Dry matter and ash were analyzed from the postfermentative samples. The pH of the contents from these bottles was also recorded. The correlation in time between fermentation end products and cumulative gas produced was determined. The results showed that the prolongation of fermentation to 72 h, especially in the case of fast-fermenting inulin and lactulose, may lead to a different end product profile (P < 0.001) compared with the profile observed at the time at which most of the substrate has disappeared. Therefore, we concluded that the fermentation product profile at the end of in vitro fermentation at a specific time point cannot be used to compare fermentability of carbohydrate sources with different fermentation kinetics in terms of gas production.

Antioxidants may be utilised for two main purposes, to protect the sensory and nutritive quality of the food and/or to protect the body against chronic and age-related diseases. Generally, antioxidants are subject to process degradation and, when given to the body in their free form, cannot pass cell membranes and are rapidly cleared from the general circulation. Because of their unique properties, lipid-based nanoencapsulation systems enhance the performance of antioxidants by improving their solubility and bioavailability, in vitro and in vivo stability, and preventing their unwanted interactions with other food components. This paper reviews nanoliposomes, archaeosomes and nanocochleates with respect to their potential applications as antioxidant carriers in foods.

An experiment was conducted to examine differences in the in vitro fermentability of four carbohydrate-rich feed ingredients and two weaning piglet diets with and without these ingredients, using both the ileal contents and the faeces of unweaned piglets as inocula. In the first part of the experiment, cumulative gas production was measured over time, using faecal inocula mixed from nine specially raised crossbred piglets (no creep feed or antibiotics) at 3 weeks of age. Inulin, lactulose, unmolassed sugar beet pulp, wheat starch and the two diets were used as substrates and fermented in vitro for 72 h. Gas production was measured as an indicator of the kinetics of fermentation. Fermentation end-products, including volatile fatty acids and ammonia, and organic matter loss, were also measured. For the fermentations of feed ingredients, samples were also collected for polymerase chain reaction/denaturing gradient gel electrophoresis analyses initially and after the fermentation process, to study changes in the composition of the bacterial community. This procedure was repeated 1 week later, using ileal contents from the same piglets as inoculum. There were significant differences between the inocula, in terms of both overall fermentation characteristics and composition and between the substrates. There was also a significant interaction between inocula and substrates, which suggests that there were potentially important differences in the microbial activity occurring in these two areas of the gastrointestinal tract (GIT). For the two diets, one with and one without addition of these fermentable ingredients, there were significant differences in terms of the kinetics, but less so in terms of the end-products of fermentation. It was concluded that inocula from both the small and large intestine should be used to obtain a more accurate assessment of potential feed ingredients which will stimulate fermentation in the piglet GIT. Copyright © 2005 Society of Chemical Industry

Recently, it has become apparent that GIT fermentation is not only of interest for ruminant animals, but also for monogastrics. While it is now widely accepted that the fermentation process and its resultant end-products can have important influences on animal health, little is known about the microbiological and immunological processes involved. In terms of animal health, most interest at the moment is focussed on those moments in animals' lives when they are faced with sudden changes resulting in stress. The period of weaning in piglets is a typical example of this. The most easily accomplished and appropriate way to influence GIT fermentation processes is that of dietary intervention. This is reflected by the widespread interest in so-called pre- and pro-biotics. Given the complexities of the interactions occurring in the animal itself, it is hardly surprising that in vitro techniques are being widely used: firstly to examine potential substrates for their fermentability and possible inclusion in diets, and secondly, to assess changes in the microbial populations in response to these substrates. This paper will review the techniques currently in use for these two aspects of monogastric fermentation, and provide examples of their use.

The aim of this study was to test the hypothesis of an improved gut environment of post-weaning piglets when administered a blend of essential oils (EO; thymol and cinnamaldehyde) and an enzyme combination (xylanase and β-glucanase (XB)) either alone or in combination. To assess the effect of dietary treatments, faecal nutrient digestibility and microbial counts, as well as ileum histology and gene expression of inflammatory mediators were evaluated. One hundred and ninety-two weaned piglets were allocated into four experimental treatments, and fed the basal diet (CTRL) either without or with EO, XB or their combination (EO+XB) for a 42-day period. The experiment concerning digestibility was designed with two periods (period I: days 15 to 21; period II: days 29 to 35) and the faeces were collected on days 20, 21, 34 and 35. On day 42, six piglets from each treatment were slaughtered. It was found that EO, XB and EO+XB supplementation did not affect (P>0.05) the growth performance of the piglets from days 0 to 42. Moreover, no dietary effect on faecal score was observed. Faecal digestibility of dry matter, organic matter, ash, dietary fibre, lipid, CP and NDF were increased from period I to period II (P0.05) of EO, XB or their combination on the faecal digestibility was observed at both periods. Compared with the CTRL diet, dietary XB reduced the faecal Lactobacillus and Escherichia coli counts but increased the Lactobacillus to Coliforms ratio on day 42 (P=0.02, 0.03 and 0.03, respectively), and all the additives supplementations decreased the counts of faecal Coliforms on day 42 (P

SummaryThe objective of the study was to test the effect of a direct fed microbial (DFM) on the performance of broilers compared to an antibiotic growth promoter under large scale, commercial production settings. Three dietary treatments were tested in a completely randomized design including: 1) a control (C) diet containing 500 FTU/kg phytase and a mixture of xylanase, amylase, protease ; 2) C+ a specific three-strain combination of Bacillus spp. (DFM) and 3) C+ bacitracin methylene disalicylate (BMD). Six, similar commercial broiler houses (15,300 birds per house) were used to give two replicate houses per treatment. The birds (Hubbard x Cobb500) were fed pelleted and crumbled diets ad libitum throughout the 44 day trial period. Due to the large scale, commercial nature of the trial, no significant differences were observed in production parameters among treatments, except that DFM treatment resulted in significantly lower mortality numbers in the last two days (43 to 44d) compared to the control. However, the DFM treatment group showed numerically higher live bodyweight, lower feed conversion ratio (corrected for body weight and mortality) and lower total mortality weight compared to either the control or BMD groups, resulting in an improved production efficiency factor. When compared to control, using DFM resulted in a gross benefit of US$ 0.06 /bird, while using BMD was not cost effective. In conclusion, DFM containing a three-strain combination of Bacillus spp. may be used as an alternative to antibiotic growth promoters, resulting in economic benefit under commercial production settings in broilers fed commercial diets.

The validation of a dual in vivo-in vitro digestibility assay ('dual digestibility assay') for separately predicting the upper-tract, hindgut and total tract digestibility of nutrients in humans, as estimated using organic matter digestibility (OMD), is described. Human upper-tract OMD was predicted using an animal (rat) model with digesta from the terminal ileum collected from rats fed one of four complete human diets (wheat bran diet, pectin diet, mixed low-fibre diet, mixed high-fibre diet). Large intestinal OMD was predicted using an in vitro hindgut fermentation assay employing a human faecal inoculum and with the rat ileal digesta as the substrate.A comparison of total tract OMD of the four diets from a human balance study (OMDhuman ) with that predicted using the dual digestibility assay (OMDdual ) showed no significant differences (P0.05). OMDhuman and OMDdual were highly correlated (r = 0.953, P = 0.047).The dual digestibility assay accurately predicts the uptake of dietary nutrients (as grams of organic matter) in humans over the total tract. The assay is able to separately quantify the digestibility of nutrients in the upper and lower digestive tracts. The validation of the dual digestibility assay needs to be extended to a wider range of human diets.

BACKGROUND: Proton nuclear magnetic resonance (NMR)-based metabonomics has only recently been applied to nutritional research. The limitation of any analytical technique is its sensitivity in detecting the smallest variation. Alterations in nutrition often produce only subtle metabolic modulations. The objective of this study was to determine if NMR-based metabonomics could detect variations in the metabolic profile of urine from pigs digesting either native casein (NC) or the same casein that had been enzymatically hydrolysed (EHC). NMR permits simultaneous detection of a large number of metabolites, thus allowing detection of unanticipated metabolic fluctuations that may otherwise have gone undetected with the use of only targeted analysis. RESULTS: Partial least squares discriminant analysis identified significantly (P < 0.05) higher urinary excretions of leucine, valine, taurine and glycine by pigs on the EHC-based diet. CONCLUSION: NMR-based metabonomics is a sensitive method that can uncover unanticipated metabolic changes brought about by physicochemical changes to the feedstock (i.e. hydrolysis). The data show a lower efficiency of retention by the kidney of some amino acids following ingestion of a hydrolysed protein. Copyright © 2012 Society of Chemical Industry

The aim was to optimise inoculum concentration and incubation duration for a published in vitro hindgut digestibility assay using ileal digesta (sampled from the chicken or rat) pertaining to a mixed human diet as the substrate. The study also sought to investigate the digestibility of the inoculum itself and the importance of correcting for this in the in vitro hindgut digestion assay. For two assays, hindgut dry matter digestibility (DMD) generally increased with inoculum concentration. A sharp increase in DMD observed at high inoculum concentrations may have been related to problems with filtering the inoculum. An inoculum concentration of 160 g/L was considered optimal based on close agreement of observed values with previously published in vivo hindgut dry matter digestibility for similar diets. One of the methods was chosen for optimisation of the duration of incubation. Ileal substrate organic matter digestibility (OMD) increased with increasing time of incubation for all diets. An incubation duration of 18 h using a mean inoculum digestibility value for calculation purposes was considered optimal based on observed in vivo hindgut DMD values in humans, but there was little difference in estimated in vitro hindgut DMD between 18 and 24h incubation durations. Although considerably lower than the OM digestibility of the substrate (no less than 51% after 48 h), the OM digestibility of the inoculum (13% after 48 h) itself was of significance in calculating estimated digestibility. The optimised assay gave realistic hindgut OMD values ranging from 55% to 79% (Wheat Bran Diet and Pectin Diet, respectively) using an 18-h incubation duration.

Neonatal diarrhoea in newborn piglets is an important problem in pig production that is frequently diagnosed as being the result of Clostridium perfringens infections. During parturition and in the first hours of life, the sow transmits the pathogen to its offspring. The objective of this study was to examine the possible prophylactic effect of a drinking water supplement containing organic acids and medium chain fatty acids (Selko-4-Health © ), administered to sows on prevalence of neonatal diarrhoea in piglets during early lactation. The study was carried out at a farm with 1300 sows with a high incidence of neonatal diarrhoea. Gestating sows received the water supplement (0.1% per litre) daily from day 35 to end of gestation and during the lactation phase for 2 days a week. Excreta of sows were collected at day 0, 35, 56, 91 of gestation and at day 21 of lactation for microbial examination counting the numbers of Lactobacilli and Clostridium spp.. The number and type of veterinary treatments were recorded during the trial period. The numbers of Clostridium spp. in faecal samples of sows decreased progressively from day 0 to 91 in gestation from log 6 to log 4 cfu/g. Counts in faecal samples of lactating sows (day 21) decreased from log 6 to log 3 cfu/g. There was a pronounced decrease in the ratio Clostridium spp to Lactobacilli spp., indicating a more specific effect of the water supplements towards lowering Clostridium spp. counts whilst maintaining higher levels of Lactobacilli. Although there was no effect on mortality of piglets, the number of veterinary treatments of newborn piglets decreased during the trial period, leading to a total reduction in antibiotic usage of 60%. The improved health status of neonatal piglets was also associated with a reduction of meningitis incidence.

Antibiotic treatment of piglets post-weaning may lead to re-occurring diarrhoea after stopping the antibiotic treatment. The objective of the study was to test the efficacy of a commercial drinking water supplement containing organic acids and medium chain fatty acids (Selko-4-Health © ) on diarrhoea control in piglets weaned at 26 days of age. In total 244 piglets were allocated at weaning to 4 treatments in a 2x2 experimental design for the duration of 4 weeks. Piglets received either a non-medicated feed, oxytetracycline medicated feed (400 ppm) during the first week post weaning and thereafter no medication, a drinking water supplement during the whole experimental period or the combination of the two treatments. Jejunal samples were taken of 4 piglets from each treatment at 2 and 4 weeks post-weaning to examine the intestinal microbiota with 16SRNA gene-targeted Denaturant Gradient Gel Electrophoresis, quantitative PCR and the Pig Intestinal Tract Chip (PITChip), a diagnostic microarray custom-designed for the profiling of porcine intestinal microbiota. None of the treatments significantly affected performance. Both, the antibiotic treatment as the water supplement treatment significantly lowered the incidence of diarrhoea in week 2 and 3 post-weaning and in the overall experimental period (p

Protein is often considered the most satiating macronutrient. The objective was to determine the short-term effect of mixtures of whey protein and glycomacropeptide (GMP) versus a carbohydrate control on satiety in healthy adult humans. The study was a randomised crossover Latin Square design. On 4 separate days, fifty healthy subjects (19 males and 31 females) received a subject-specific breakfast (08:00 h), a preload drink (12:00 h) and lunch (12:30 h). The preload drink was presented as a milkshake with either maltodextrin carbohydrate (control), whey protein isolate (WPI) with no GMP, WPI with naturally present 21% GMP or WPI with naturally present 21% GMP plus added GMP. Satiety was assessed using visual analogue scales (VAS) and by determining ad libitum food intake during a cafeteria style meal offered 30 min after the preload. The VAS indicated that the lower GMP treatment induced a greater feeling of fullness immediately after consumption of the preload compared with the other treatments. Energy and macronutrient intake at lunch did not differ significantly ( p > 0.05) between treatments although subjects chose to eat foods higher in carbohydrate and lower in protein after the protein preloads. Women consumed the least amount of protein after the protein preloads whereas no difference was found in men. There was some evidence that whey proteins and their components enhance satiety over a short-term period compared to carbohydrate but there was no consistent effect of either whey protein alone or glycomacropeptide.

Pre-weaning development of microbial activity has an effect on post-weaning establishment of the gastro-intestinal tract (GIT) microbiota. An in vivo study was conducted, to evaluate the effect of age on fermentation end-product profiles during the post-colostrum suckling period, as the variation in composition of mature milk is minimum. Sixteen piglets from two litters (eight per litter) were selected. During the study, piglets had free access to sow's milk, but no creep feed, nor antibiotic treatments. Two piglets from each litter were sacrificed on d 11, 18, 25 and 32 of age. The digesta samples were collected from the beginning and end of the small intestine, caecum and colon. Samples were analyzed for fermentation end-product concentrations. Combining the results from all the GIT sites, it was observed that, total VFA concentration increased with age of the piglets. There was a significant rise in acetic acid concentrations, with a significant decrease in lactic acid concentrations from d11 to d32, while the proportions of SCFA, (acetic acid not, vert, similar 72%, propionic acid not, vert, similar 15% and butyric acid not, vert, similar 6% of total VFA) and ammonia concentrations remained unchanged. These results clearly suggest that, the microbial activity in terms of fermentation end-product profile skewed from lactic acid to acetic acid as a major product during the post-colostrum suckling period. This may be attributed to lower substrate availability due to increased number of microbes or increased diversity in the microbiota in time.

Increasing the dietary amount of fermentable carbohydrates (FC) may counteract the negative effects of protein fermentation in newly weaned piglets. To study this hypothesis, 272 newly weaned piglets were allotted to 4 dietary treatments in a 2 × 2 factorial arrangement with low and high FC (7.5 and 13.5%) and crude protein (CP, 15 and 22%) content as respective factors. Intestinal histology, enzyme activity, microbiota and fermentation products were determined in 8 pigs per treatment 7 days post-weaning. In the 4 wk experimental period, interactions between dietary CP and FC content were found for feed intake ( P = 0.022), daily gain ( P = 0.001), and gain:feed ( P = 0.033). The high-FC content reduced daily gain by 50 g/d in the low-CP diet, whereas the FC content did not affect growth performance in the high-CP diet. Over the 4 wk experimental period, daily gain (350 g/d) and feed intake (519 g/d) were highest for piglets on the low-CP low-FC diet. The high-FC content resulted in an increase in number of lactobacilli ( P = 0.047) and a decrease of total coliforms ( P = 0.06) in the small intestine. It increased the lactic acid content ( P = 0.08) and reduced the ammonia content ( P = 0.04) in the small intestine and increased the VFA content in the colon ( P = 0.009). The reduction in CP content reduced ammonia concentration in the small intestine ( P = 0.003). We concluded that dietary FC influenced microbial population and fermentation products in the gut. However, this was not reflected in an increased growth performance.

An in vivo experiment was conducted to monitor the changes in fermentation end products in the feces of weaning piglets due to the inclusion of selected fermentable carbohydrates in the diet. The experiment involved 3 groups of 16 piglets each. Specially raised piglets (neither antibiotics nor creep feeding) were weaned abruptly at 4 wk of age. The piglets were offered 1 of 2 dietary treatments [a control diet (CON), or a fermentable carbohydrate-enriched diet (CHO)] and were subjected to 1 of the 2 fasting treatments (fasting for 2 d at the beginning of the experimental period or nonfasting). Fecal samples were collected per rectum every day during the experimental period. Piglets were slaughtered at the end of the 10-d experimental period, and digesta samples were collected from different parts of the gastrointestinal tract (GIT): the first half of the small intestine, the second half of the small intestine, the cecum, and colon. The DM, VFA profile, and ammonia concentrations were analyzed from the fecal and digesta samples. Daily feed intake was also recorded. There was no difference in concentrations of VFA in feces between the treatment groups. Ammonia concentration was lower (P < 0.05) in piglets fed the CHO diet compared with those fed the CON diet in both feces and digesta from different parts of GIT. Fasting had no effect on fermentation end products in feces. This study demonstrated that the inclusion of fermentable carbohydrates in weanling diets reduces protein fermentation along the GIT and also reduced the fecal concentration of ammonia.

The in vitro cumulative gas production technique can be used to assess microbial activity of a complex community, in relation to fermentation of a particular energy source. Therefore, in combination with an in vivo study to examine the effects of two different diets for weaning piglets, microbial activities of faeces were compared from animals on the two different diets. The two diets were: CHO diet [containing added fermentable carbohydrates, including sugarbeet pulp (SBP) and wheat starch (WST)], and control diet without any added fermentable carbohydrates. Neither diet contained antibiotics or extra added copper. Twenty-four piglets were selected from 12 litters (two per litter), weaned at 4 weeks of age (neither creep feeding nor any antibiotic treatment before and during the study), and introduced to one of the two diets. After 9 days on the diet, faecal samples were collected from selected animals, and tested for their activity in terms of gas production kinetics, and end-products such as volatile fatty acids, ammonia and dry matter disappearance of the two test substrates SBP and WST. The bacterial diversity was also analysed before and after in vitro fermentation using denaturing gradient gel electrophoresis analysis of amplified 16S rRNA genes. There were differences both in kinetics and end-products of the substrates. More interestingly, significant differences were detected between inocula, although mainly in terms of fermentation kinetics of the two substrates. With the CHO inoculum, SBP was fermented faster than with the control, while this effect was reversed for WST. Significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the microbial community enrichment on SBP as compared with WST at the end of fermentation. The difference between the kinetics of SBP compared with WST fermentation by faecal microbiota from the CHO diet fed piglets suggests better adaptation to SBP fermentation than to WST fermentation. The WST fermentation was more unexpected, given that a significant amount of starch is known to be fermentable by the small intestinal microbiota. It was concluded that the microbial community composition and activity in the GIT may be changed in response to diet, and that this change can be detected in vitro. Copyright © 2005 Society of Chemical Industry

Using 16S rRNA gene-based approaches, we analyzed the responses of ileal and colonic bacterial communities of weaning piglets to dietary addition of four fermentable carbohydrates (inulin, lactulose, wheat starch, and sugar beet pulp). An enriched diet and a control diet lacking these fermentable carbohydrates were fed to piglets for 4 days ( n = 48), and 10 days ( n = 48), and the lumen-associated microbiota were compared using denaturing gradient gel electrophoresis (DGGE) analysis of amplified 16S rRNA genes. Bacterial diversities in the ileal and colonic samples were measured by assessing the number of DGGE bands and the Shannon index of diversity. A higher number of DGGE bands in the colon (24.2 ± 5.5) than in the ileum (9.7 ± 4.2) was observed in all samples. In addition, significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the colonic microbial community of weaning piglets fed the fermentable-carbohydrate-enriched diet for 10 days than in the control. Selected samples from the ileal and colonic lumens were also investigated using fluorescent in situ hybridization (FISH) and cloning and sequencing of the 16S rRNA gene. This revealed a prevalence of Lactobacillus reuteri in the ileum and Lactobacillus amylovorus -like populations in the ileum and the colon in the piglets fed with fermentable carbohydrates. Newly developed oligonucleotide probes targeting these phylotypes allowed their rapid detection and quantification in the ileum and colon by FISH. The results indicate that addition of fermentable carbohydrates supports the growth of specific lactobacilli in the ilea and colons of weaning piglets.