Functional amino acid supplementation attenuates the negative effects of plant-based nursery diets on the response of pigs to a subsequent Salmonella Typhimurium challenge.

Functional amino acids (FAA) attenuate the effects of Salmonella challenge in pigs. However, this may be affected by protein source (PS). The objective of the present study was to determine the effects of nursery dietary PS and FAA supplementation on growth performance and immune status of pigs subsequently challenged with Salmonella Typhimurium (ST). Thirty-two weanling pigs (8.7 ± 0.23 kg) were assigned to a feeding program for 31 d in a 2 × 2 factorial arrangement. Factors were dietary PS (plant-based [PB] vs. animal-based [AB]) and FAA profile (basal [FAA-] or supplemented [FAA+; Thr, Met, and Trp at 120% of requirements]). Pigs were subsequently placed on a common grower diet and, after a 7-d adaptation, were inoculated with ST and monitored for 7 d postinoculation. Growth performance, rectal temperature, fecal score, gut health, ST shedding score, intestinal colonization and translocation, and blood parameters of acute-phase response and antioxidant balance were measured pre- and postinoculation. Data were analyzed with a 2 (AB vs. PB) × 2 (FAA- vs. FAA+) factorial arrangement of treatments and differences between means were considered significant at P ≤ 0.05. Postinoculation fecal score was worse, ST shedding, cecal myeloperoxidase, and cecal and colonic ST colonization were greater in PB compared to AB pigs (P < 0.05). Translocation of ST to spleen was decreased by FAA+ (P < 0.05), regardless of dietary PS. Postinoculation, AB pigs had greater average daily gain compared to PB-FAA- (P < 0.05). Pigs fed AB-FAA- showed increased average daily feed intake compared to PB-FAA- pigs (P < 0.05) and feed efficiency was increased in AB-FAA+ compared to PB-FAA- pigs (P < 0.05). Feeding PB ingredients in nursery diets seems to increase susceptibility of pigs to Salmonella. Moreover, FAA supplementation partially attenuated the negative effects of PB diets on the response of pigs to ST challenge.

While long-term growth performance of weaned pigs is not negatively affected by feeding nursery diets containing only plant-based protein sources, these pigs may be more susceptible to subsequent disease challenges. It has been previously shown that supplementation with key functional amino acids improves growth performance and the immune status of pigs during intestinal pathogen challenge. A study was performed to determine the effect of feeding nursery diets containing only plant-based protein sources or including animal-based protein sources with or without supplementation with a blend of functional amino acids (methionine, threonine, and tryptophan) on growth and immune status during a subsequent Salmonella challenge. Pigs fed diets containing animal-based protein sources had improved growth performance and immune status compared to pigs fed plant-based diets, regardless of the inclusion of functional amino acids. Pigs fed plant-based diets were more susceptible to the disease challenge, however, this was partially mitigated by the inclusion of functional amino acids. The inclusion of animal-based protein sources may be necessary to optimize pig health and performance, however, functional amino acid inclusion may be beneficial when plant-based diets are fed.

Ileal alkaline phosphatase is upregulated following functional amino acid supplementation in Salmonella Typhimurium-challenged pigs.

We recently showed that functional amino acid (FAA) supplementation improves growth performance and immune status of Salmonella Typhimurium (ST)-challenged pigs, which was further improved by a longer adaptation period. It is expected that the effects are associated with increased activity of intestinal alkaline phosphatase (IAP). The objective of this study was to evaluate the effects of FAA supplementation and adaptation period on the ileal, cecal, and colonic activity of IAP in weaned pigs challenged with ST. In experiment 1, a total of 32 mixed-sex weanling pigs were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement with low (LP) or high protein (HP) content and basal (FAA-) or FAA profile (FAA+; Thr, Met, and Trp at 120% of requirements) as factors. In experiment 2, a total of 32 mixed-sex weanling pigs were randomly assigned to one of four dietary treatments, being FAA- fed throughout the experimental period (FAA-) or an FAA profile fed only in the post-inoculation (FAA + 0), for 1 wk pre- and post-inoculation (FAA + 1), or throughout the experimental period (FAA + 2). In experiments 1 and 2, after a 7- and 14-d adaptation period, respectively, pigs were inoculated with saline solution containing ST (3.3 and 2.2 × 109 CFU/mL, respectively). Plasma alkaline phosphatase was measured on days 0 and 7 post-inoculation in experiment 1, and IAP (ileum, cecum, and colon) was measured in experiments 1 and 2. Correlations among ileal IAP and serum albumin and haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), and reduced:oxidized glutathione, ileal myeloperoxidase, ST shedding and ileal colonization, and post-inoculation average daily gain, feed intake (ADFI), and gain:feed were also analyzed. In experiment 1, plasma alkaline phosphatase was decreased with ST inoculation and the overall content was increased in LP-FAA+ compared with LP-FAA- (P < 0.05). Moreover, ileal IAP was increased in FAA+ compared with FAA- pigs in both studies (P < 0.05) regardless of adaptation time (P > 0.05). IAP was positively correlated with MDA and ADFI and negatively correlated with SOD and ST shedding in experiment 1 (P < 0.05). These results demonstrate a positive effect of FAA supplementation, but not adaptation period, on ileal alkaline phosphatase activity in Salmonella-challenged pigs, which may be associated with improvements in antioxidant balance.

Functional amino acid (FAA) supplementation has been shown to improve gut health and antioxidant defense in weaned piglets challenged with Salmonella Typhimurium (ST), regardless of dietary protein content. The beneficial effects were further improved when pigs were adapted to FAA for 2 wk prior to the ST challenge. Recent evidence has shown that intestinal alkaline phosphatase (IAP), which may be influenced by nutritional factors, attenuates intestinal inflammation, possibly due to gut microbiota modulation. This study is the first to identify that ileal IAP activity is increased following FAA supplementation in ST-challenged pigs, regardless of adaptation period. Moreover, ileal IAP activity correlated with systemic markers of antioxidant defense, which highlights the enzyme's role in attenuating systemic infection. Overall, the development of feeding strategies with positive effects on IAP activity is of interest, due to the enzyme's central role on the gut and whole-body homeostasis and health.

Effects of exogenous ß-glucanase on ileal digesta soluble ß-glucan molecular weight, digestive tract characteristics, and performance of coccidiosis vaccinated broiler chickens fed hulless barley-based diets with and without medication.

INTRODUCTION: Limited use of medication in poultry feed led to the investigation of exogenous enzymes as antibiotic alternatives for controlling enteric disease. The objective of this study was to evaluate the effects of diet ß-glucanase (BGase) and medication on ß-glucan depolymerization, digestive tract characteristics, and growth performance of broilers. MATERIALS AND METHODS: Broilers were fed hulless barley (HB) based diets with BGase (Econase GT 200P from AB Vista; 0 and 0.1%) and medication (Bacitracin and Salinomycin Na; with and without) arranged as a 2 × 2 factorial. In Experiment 1, 160 broilers were housed in cages from d 0 to 28. Each treatment was assigned to 10 cages. In Experiment 2, broilers (2376) were housed in floor pens and vaccinated for coccidiosis on d 5. Each treatment was assigned to one floor pen in each of nine rooms. RESULTS: In Experiment 1, the soluble ß-glucan weighted average molecular weight (Mw) in the ileal digesta was lower with medication in the 0% BGase treatments. Peak molecular weight (Mp) and Mw were lower with BGase regardless of medication. The maximum molecular weight for the smallest 10% ß-glucan (MW-10%) was lower with BGase addition. In Experiment 2, Mp was lower with medication in 0% BGase treatments. Beta-glucanase resulted in lower Mp regardless of medication, and the degree of response was lower with medication. The MW-10% was lower with BGase despite antibiotic addition. Body weight gain and feed efficiency were higher with medication regardless of BGase use through-out the trial (except d 11-22 feed efficiency). Beta-glucanase resulted in higher body weight gain after d 11 and worsened and improved feed efficiency before and after d 11, respectively, in unmedicated treatments. CONCLUSION: BGase and medication caused the depolymerization of soluble ileal ß-glucan. Beta-glucanase acted as a partial replacement for diet medication by increasing growth performance in coccidiosis vaccinated broilers.

A longer adaptation period to a functional amino acid-supplemented diet improves growth performance and immune status of Salmonella Typhimurium-challenged pigs.

We recently showed that dietary supplementation with key functional amino acids (FAA) improves growth performance and immune status of Salmonella Typhimurium (ST)-challenged pigs. It is not known if ST-challenged pigs will benefit from a longer adaptation period to FAA. The objective of this study was to evaluate the effects of different adaptation periods to diets containing FAA above requirements for growth on performance and immune response of weaned pigs subsequently challenged with ST. A total of 32 mixed-sex weanling pigs (11.6 ± 0.3 kg) were randomly assigned to 1 of 4 dietary treatments, being a basal amino acid (AA) profile fed throughout the experimental period (FAA-) or a functional AA profile (FAA+; Thr, Met, and Trp at 120% of requirements) fed only in the postinoculation (FAA+0), for 1 wk pre- and postinoculation (FAA+1), or throughout the experimental period (FAA+2). After a 14-d adaptation period, pigs were inoculated with ST (2.15 × 109 CFU/mL). Growth performance, body temperature, fecal score, acute-phase proteins, oxidant/antioxidant balance, score for ST shedding in feces and intestinal colonization, and fecal and digesta myeloperoxidase (MPO) were measured pre- and postinoculation. Postinoculation body temperature and fecal score, serum haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), and fecal MPO were increased while serum albumin and plasma reduced glutathione (GSH):oxidized glutathione (GSSG) were reduced compared to pre-inoculation (P < 0.05). Average daily gain and G:F were greater in FAA+2 pigs compared to FAA- pigs (P < 0.05). Serum albumin was higher in FAA+2 and FAA+1 compared to FAA+0 and FAA- pigs (P < 0.05) while FAA+2 pigs had lower haptoglobin compared to FAA- (P < 0.05). Plasma SOD was increased and GSH:GSSG was decreased in FAA- pigs compared to the other treatments (P < 0.05). Score for ST shedding in feces was progressively lower from d 1 to 6 regardless of treatment (P < 0.05) and was lower in FAA+2 pigs compared to FAA- and FAA+0 (P < 0.05). Counts of ST in colon digesta were higher in FAA- and FAA+0 pigs compared to FAA+2 (P < 0.05). Fecal and colonic digesta MPO were lower in FAA+2 and FAA+1 pigs compared to FAA- (P < 0.05). These results demonstrate a positive effect of a longer adaptation period to FAA-supplemented diets on performance and immune status of weaned pigs challenged with Salmonella.

Functional amino acid supplementation, regardless of dietary protein content, improves growth performance and immune status of weaned pigs challenged with Salmonella Typhimurium.

High dietary protein may increase susceptibility of weaned pigs to enteric pathogens. Dietary supplementation with functional amino acids (FAA) may improve growth performance of pigs during disease challenge. The objective of this study was to evaluate the interactive effects of dietary protein content and FAA supplementation above requirements for growth on performance and immune response of weaned pigs challenged with Salmonella. Sixty-four mixed-sex weanling pigs (13.9 ± 0.82 kg) were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement with low (LP) or high protein (HP) content and basal (AA-) or FAA profile (AA+; Thr, Met, and Trp at 120% of requirements) as factors. After a 7-d adaptation period, pigs were inoculated with either a sterile saline solution (CT) or saline solution containing Salmonella Typhimurium (ST; 3.3 × 109 CFU/mL). Growth performance, body temperature, fecal score, acute-phase proteins, oxidant/antioxidant balance, ST shedding score in feces and intestinal colonization, fecal and digesta myeloperoxidase (MPO), and plasma urea nitrogen (PUN) were measured pre- and postinoculation. There were no dietary effects on any measures pre-inoculation or post-CT inoculation (P > 0.05). Inoculation with ST increased body temperature and fecal score (P < 0.05), serum haptoglobin, plasma superoxide dismutase (SOD), malondialdehyde (MDA), PUN, and fecal MPO, and decreased serum albumin and plasma reduced glutathione (GSH):oxidized glutathione (GSSG) compared with CT pigs (P < 0.05). ST-inoculation reduced average daily gain (ADG) and feed intake (ADFI) vs. CT pigs (P < 0.05) but was increased by AA+ vs. AA- in ST pigs (P < 0.05). Serum albumin and GSH:GSSG were increased while haptoglobin and SOD were decreased in ST-inoculated pigs fed AA+ vs. AA- (P < 0.05). PUN was higher in HP vs. LP-fed pigs postinoculation (P < 0.05). Fecal ST score was increased in ST-inoculated pigs on days 1 and 2 postinoculation and declined by day 6 (P < 0.05) in all pigs while the overall score was reduced in AA+ vs. AA- pigs (P < 0.05). Cecal digesta ST score was higher in HP vs. LP-fed pigs and were lower in AA+ compared with AA- fed pigs in the colon (P < 0.05). Fecal and digesta MPO were reduced in ST pigs fed AA+ vs. AA- (P < 0.05). These results demonstrate a positive effect of FAA supplementation, with minimal effects of dietary protein, on performance and immune status in weaned pigs challenged with Salmonella.

Hulless barley and beta-glucanase levels in the diet affect the performance of coccidiosis-challenged broiler chickens in an age-dependent manner.

Diet ß-glucanase (BGase) depolymerizes viscous ß-glucan into lower molecular weight carbohydrates, which might act as a prebiotic in chickens exposed to enteric disease. Coccidiosis-challenged broiler chickens were fed graded levels of hulless barley (HB) and BGase to determine their effects on growth performance. Broilers were fed high ß-glucan HB (CDC Fibar; 0, 30, and 60% replacing wheat) and BGase (Econase GT 200P; 0, 0.01, and 0.1%) in a 3 × 3 factorial arrangement. A total of 5,346 broilers were raised in litter floor pens and vaccinated for coccidiosis in feed and water on day 5. Each treatment was assigned to 1 pen (66 birds) in each of 9 rooms. Statistical significance was set at P ≤ 0.05. Overall, HB decreased body weight gain (BWG) and increased feed: gain ratio (F:G) of broilers. From day 0 to 11, BGase did not affect BWG and F:G, at the 0 and 30% HB. However, at 60% HB, the 0.01% BGase improved them, and the 0.1% BGase had no effect on BWG and increased F:G. For the day 22 to 32 and 0 to 32 periods, BGase did not affect BWG for 0 and 30% HB levels, but for the 60% HB, both BGase levels increased gain. The 0.1% level of BGase resulted in the lowest F:G for all HB levels, with the degree of response increasing with HB. No interaction was found for ileal digesta viscosity at day 11; the level of HB did not affect viscosity, but both levels of BGase decreased viscosity. At day 33, BGase did not affect viscosity at 0 and 30% HB levels, but viscosity was lowered for the 0.1% BGase treatment at the 60% HB level. In conclusion, HB reduced broiler performance, and BGase alleviated most but not all the effects. In young birds fed 60% HB, 0.1% BGase did not impact BWG and increased F:G.

Effect of supplemental threonine above requirement on growth performance of Salmonella typhimurium challenged pigs fed high-fiber diets1.

It was shown previously that high dietary fiber (DF) and immune system stimulation (ISS) with systemic Escherichia coli lipopolysaccharide independently increased the threonine (Thr) requirement to maximize growth performance and protein deposition (PD). However, no additive effects on the Thr requirement were observed when both DF and ISS were present. The objective of the present study was to investigate whether supplementing Thr to meet previously estimated requirements for high DF and systemic immune challenge would maintain performance of pigs exposed to an enteric immune challenge when fed high DF. A total of 128 pigs (22.6 ± SD = 1.6 kg initial BW) were assigned to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement in a randomized complete block design (n = 8 pens/treatment and 4 pigs/pen) for 28 d. Treatments were a low-fiber (LF; 13% total DF) or high-fiber (HF; 20% total DF) diet with either a standard (STD; 0.65% SID) or supplemental (SUP; 0.78% SID) Thr level. After a 7-d adaptation, pigs were orally inoculated with 2 mL (2.3 × 109 CFU/mL) of Salmonella typhimurium (ST). Blood samples and rectal swabs were obtained and rectal temperature recorded to determine clinical responses and ST shedding. On day 7 postinoculation, 1 pig/pen was euthanized and mesenteric lymph nodes, spleen, and digesta (ileum, cecum, and colon) were sampled to assess ST colonization and translocation. Body weight and feed intake were recorded on day 0, 7, and 21 postinoculation to calculate ADG, ADFI, and G:F. Rectal temperature increased (P < 0.05) 24 h postinoculation and remained elevated at day 6. Serum albumin concentration decreased (P < 0.05), whereas haptoglobin concentration increased (P < 0.05) postinoculation. There was no fiber or Thr effect (P > 0.05) on ST counts in the ileum and cecum, but a fiber × Thr interaction (P < 0.05) was observed in the colon. Supplemental Thr improved (P < 0.05) growth performance in LF- and HF-fed challenged pigs. However, performance of supplemented HF challenged pigs was less than (P < 0.05) supplemented LF challenged pigs. These results suggest that Thr supplemented to meet requirements for high DF and systemic immune challenge was not sufficient to maintain growth performance of pigs fed HF diets and challenged with an enteric pathogen.

Phytate degradation in gnotobiotic broiler chickens and effects of dietary supplements of phosphorus, calcium, and phytase.

Gnotobiotic broiler chickens were used to study interactive effects of supplemented phosphorus, calcium (PCa), and phytase (Phy) on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and release of myo-inositol in the digestive tract. In 2 subsequent runs, the chickens were subjected to 1 of 4 dietary treatments with and without PCa and Phy supplementation. Sanitized eggs were hatched in 8 germfree isolators, and a minimum of 9 male Ross 308 chickens were placed in each pen (total 16 pens). Treatments implemented on day 10 included gamma-irradiated diets without (PCa-; 4.1 g P and 6.2 g Ca/kg DM) or with (PCa+; 6.9 g P and 10.4 g Ca/kg DM) monosodium phosphate and limestone supplementation and without (Phy-) or with (Phy+) 1,500 FTU Phy/kg feed in a factorial arrangement. On day 15, digesta was collected from different sections of the intestinal tract and analyzed for InsP isomers and myo-inositol. The isolators did not remain germfree, but analysis of contaminants and results of InsP degradation indicated no or minor effects of the identified contaminants. Prececal InsP6 disappearance was 42% with the PCa-Phy- treatment and 17% with PCa+Phy-. No InsP3-4 isomers were found in the digesta of the terminal ileum in PCa-Phy-. The concentration of myo-inositol in the ileal digesta from PCa-Phy- (6.1 µmol/g DM) was significantly higher than that from PCa+Phy- (1.7 µmol/g DM), suggesting rapid degradation of the lower InsP isomers by mucosal phosphatases and their inhibition by PCa. Phytase supplementation increased InsP6 disappearance and prevented inhibitory effects of PCa supplements (72% in PCa-Phy+ and 67% in PCa+Phy+). However, PCa supplementation reduced the degradation of lower InsP isomers mainly in the posterior intestinal sections in the presence of Phy, resulting in significantly lower myo-inositol concentrations. It is concluded that mucosa-derived phosphatases might significantly contribute to InsP6 degradation in broiler chickens. The potential of mucosa-derived phosphatases to degrade InsP6 and lower InsP is markedly reduced by dietary PCa supplementation.

Supplementation with live yeast increases rate and extent of in vitro fermentation of nondigested feed ingredients by fecal microbiota.

Two studies were conducted to investigate the effect of live yeast (LY) on the in vitro fermentation characteristics of wheat, barley, corn, soybean meal (SBM), canola meal, and distillers dried grains with solubles (DDGS). In Study 1, LY yeast was added directly to in vitro fermentations inoculated with feces from lactating sows, whereas as in study 2, feces collected from lactating sows fed LY as a daily supplement was used. Selected feedstuffs were digested and the residue added to separate replicated (n = 3) fermentation reactions. Study 1 was conducted in two blocks, whereas study 2 was conducted using feces collected after a period of 3 (Exp. 1) or 4 wk (Exp. 2) of LY supplementation. Accumulated gas produced over 72 h was modeled for each substrate and the kinetics parameters compared between LY and control groups. The molar ratio of the volatile fatty acids (VFAs) produced in vitro were also compared at 12 and 72 h of incubation. In study 1, in vitro addition of yeast increased (P < 0.001) the rate of gas production (Rmax). However, a yeast × substrate effect (P < 0.05) observed for total gas accumulated (A), time to half asymptote (B), and time required to reach maximum rate of fermentation (Tmax) suggested that yeast-mediated increases in extent and rate of fermentation varied by substrate. Greater total gas production was observed only for corn and SBM, associated with greater B and Tmax. Supplementation with LY appeared to increase A and Rmax although with variation between experiments and substrates. In Exp. 1, LY decreased (P < 0.05) B and Tmax. However, a yeast × substrate effect (P < 0.05) was observed for only A (for wheat, barley, corn, and corn DDGS) and Rmax (wheat, barley, corn, and wheat DDGS). In Exp. 2, LY increased (P < 0.0001) A and decreased B. However, an interaction (P < 0.05) with substrates was observed for Rmax (except SBM) and Tmax. With exception of the DDGS samples, LY supplementation increased (P < 0.05) VFA production at 12 and 72 h of incubation. Yeast increased (P < 0.05) the molar ratios of acetic acid and branch-chain fatty acids at 12 h of incubation; however, this response was more variable by substrate at 72 h. In conclusion, LY supplementation increased the rate and extent of in vitro fermentation of a variety of substrates prepared from common feedstuffs. Greater effects were observed when LY was fed to sows than added directly in vitro, suggesting effects on fermentation were not mediated directly.

Effect of barley and oat cultivars with different carbohydrate compositions on the intestinal bacterial communities in weaned piglets.

This experiment was aimed at comparing the intestinal microbial community composition in pigs fed hulled common barley supplemented with isolated barley mixed-linked beta-glucan, four hulless barley varieties and breeding lines with mixed-linked beta-glucan contents ranging from 41 to 84 g kg(-1) and different amylose/amylopectin ratios as well as two oat varieties. Seventy-two weaned piglets were allocated to one of nine diets composed of 81.5% cereal, 6% whey, 9% soy protein isolate and 3.5% minerals. After 15 days, pigs were sacrificed and ileum and colon contents were collected for quantitative real-time PCR (qPCR) and denaturing gradient gel electrophoresis to evaluate microbial communities. Shifts in intestinal microbial communities were observed with the hulless barley cultivars with a normal to high beta-glucan content and from normal starch toward either high-amylopectin or high-amylose starch. These hulless barleys had the lowest (P<0.05) microbial diversity, whereas oats had intermediate diversity compared with low-beta-glucan hulless cultivars and hulled varieties. Furthermore, hulless varieties favoured xylan- and beta-glucan-degrading bacteria whereas mixed-linked beta-glucan-supplemented hulled barley favoured lactobacilli. Numbers of lactobacilli decreased in the ileum of pigs fed hulless/high mixed-linked beta-glucan barley-based diets. Thus, cultivar differences in both the form and the quantity of carbohydrates affect gut microbiota in pigs, which provides information for future feeding strategies.

Dietary encapsulated glycine influences Clostridium perfringens and Lactobacilli growth in the gastrointestinal tract of broiler chickens.

Three experiments were conducted to determine whether there is a causative relation between dietary glycine concentration and intestinal Clostridium perfringens growth in broiler chickens. Expt. 1 showed that glycine concentrations were higher (P < 0.05) in jejunum and ileum of birds fed fat-encapsulated glycine compared with crystalline glycine. In Expt. 2, 2 cages of 6 birds were assigned to 1 of 6 experimental diets formulated to contain 7.6 and 10.6, 17.8 and 40.6, 27.8 and 30.6, 37.8 and 20.6, 47.7 and 10.6, and 7.8 and 50.6 g/kg total glycine and proline, respectively, provided primarily by supplementation with encapsulated glycine or proline as required. In Expt. 3, 12 groups of 6 birds were fed 4 different diets supplemented with encapsulated glycine to achieve 7.6, 21.0, 34.3, or 47.7 g/kg total glycine. The birds were orally challenged with C. perfringens type A on d 1 and d 14-21 and killed on d 28. In Expt. 2, C. perfringens populations were higher (P < 0.05) in ileum and cecum of birds, which received either 37.8 or 47.7 g/kg total glycine compared with those fed 7.6 g/kg glycine. In Expt. 3, C. perfringens numbers were higher (P < 0.05) in ileum of birds fed either 34.3 or 47.7 g/kg dietary glycine than those given either 7.6 or 21.0 g/kg glycine. Conversely, lactobacilli counts in ileum and cecum were significantly lower in birds fed the higher levels of glycine in both experiments. High C. perfringens colonization and high intestinal lesion scores were associated with reduced performance (P < 0.05). We conclude that glycine is an important determinant of C. perfringens growth in the intestinal tract of broiler chickens.

Characterization of intestinal microbiota and response to dietary virginiamycin supplementation in the broiler chicken.

The inclusion of antibiotic growth promoters, such as virginiamycin, at subtherapeutic levels in poultry feeds has a positive effect on health and growth characteristics, possibly due to beneficial effects on the host gastrointestinal microbiota. To improve our understanding of the chicken gastrointestinal microbiota and the effect of virginiamycin on its composition, we characterized the bacteria found in five different gastrointestinal tract locations (duodenal loop, mid-jejunum, proximal ileum, ileocecal junction, and cecum) in 47-day-old chickens that were fed diets excluding or including virginiamycin throughout the production cycle. Ten libraries (five gastrointestinal tract locations from two groups of birds) of approximately 555-bp chaperonin 60 PCR products were prepared, and 10,932 cloned sequences were analyzed. A total of 370 distinct cpn60 sequences were identified, which ranged in frequency of recovery from 1 to 2,872. The small intestinal libraries were dominated by sequences from the Lactobacillales (90% of sequences), while the cecum libraries were more diverse and included members of the Clostridiales (68%), Lactobacillales (25%), and Bacteroidetes (6%). To assess the effects of virginiamycin on the gastrointestinal microbiota, 15 bacterial targets were enumerated using quantitative, real-time PCR. Virginiamycin was associated with increased abundance of many of the targets in the proximal gastrointestinal tract (duodenal loop to proximal ileum), with fewer targets affected in the distal regions (ileocecal junction and cecum). These findings provide improved profiling of the composition of the chicken intestinal microbiota and indicate that microbial responses to virginiamycin are most significant in the proximal small intestine.