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1. The objective of this study was to investigate the potential immunomodulatory effects of yeast (Saccharomyces cerevisiae) and yeast-derived products treated with a cell wall lytic enzyme mixture on the gene expression of toll-like receptors and cytokines of chicken B cell line (DT 40) stimulated with lipopolysaccharide. 2. The effect of brewer's yeast (Y), yeast cell wall (YCW), distilled dried grains with solubles (DDGS) and a processed yeast + nucleotide-rich product (PY/N) treated with a yeast cell wall lytic enzyme (E) was assessed using a chicken B cells and LPS challenge model. 3. Relative gene expression of toll-like receptors (TLRs) and cytokines was investigated. Treatment of cells with Y, YCW, YCW + E and PY/N upregulated the expression of TLR2b following LPS challenge. Gene expression of TLR4 was downregulated in E, YCW and YCW + E treatments compared to control, while adding DDGS and PY/N upregulated the expression of TLR4 either before or after enzyme treatment. Following LPS challenge, expression of IL-4 and IL-10 was upregulated in cells treated with YCW and PY/N, both pre and post enzyme incubation. Adding YCW and PY/N to the cells challenged with LPS upregulated the expression of IFN-γ and IL-12 before and after enzyme treatment. Treatment of cells with YCW, DDGS and PY/N increased the expression of IL-6 prior to LPS challenge. 4. In conclusion, the results suggested that yeast-derived products affected immunomodulatory activities by changing the expression of cytokines involved in the innate immune response. [ABSTRACT FROM AUTHOR]

An experiment was carried out to evaluate the short-term effect of supplementing a nucleotide-rich yeast extract (NRYE) on growth performance, gut structure, immunity and microflora of piglets raised under sanitary and unsanitary conditions. A total of 84, 21-day old piglets were used in this study; 42 piglets were raised in a room designated as the clean room that was washed once per week, whereas the other 42 piglets were raised in a room designated as the unclean room in which 7 kg of manure from the sow herd was spread on each pen floor on day 1 and 7 and the room was not washed throughout the experiment. The pigs were fed a corn–soybean meal-based diet without or with 0.1% NRYE. Each treatment had 7 replicate pens in each room, and each pen housed 3 pigs. Feed disappearance and BW were recorded on day 1 and 14. On day 14, one pig per pen was euthanized to collect ileum, mesenteric lymph nodes and spleen tissues, and cecum and colon digesta. Overall, NRYE supplementation did not affect growth performance in both clean and unclean conditions, improved kidney weight in both clean (P=0.0002) and unclean room (P<0.0001) and tended to improve the villus height/crypt depth ratio in the clean room (P=0.073). Supplementing NRYE was associated with upregulation of Ileal programmed cell death gene-1 (P=0.0003), interleukin (IL)-1β (P<0.0001), IL-6 (P=0.0003), IL-10 (P<0.0001) and tumor necrosis factor-α (TNF-α) (P<0.0001) in pigs raised in the unclean room. Supplementing the NRYE in pigs raised in the clean room suppressed growth of cecal Enterobacteriacea (P<0.0001) members and colonic Enterococcus spp. (P<0.019), improved proliferation of cecal Lactobacillus spp. (P<0.002) and colonic Clostridium cluster IV (P<0.011) and XVIa members (P<0.0002). Supplementing the NRYE in the unclean room improved proliferation of cecal Clostridium cluster IV (P<0.026) and suppressed proliferation of colonic Enterococcus spp. (P<0.037). In conclusion, supplementing the NRYE to piglets under unsanitary conditions improved ileal immune response by upregulating inflammatory cytokines, and positively modulated proliferation of beneficial gut bacteria and suppression of harmful ones in both clean and unclean rooms. [ABSTRACT FROM PUBLISHER]

This study evaluated the effect of yeast-derived products on growth performance, serum antibody levels, and mRNA gene expression of patternrecognition receptors, and cytokines in broiler chickens. Two hundred and sixteen one-day-old male broiler chickens (Ross-308) were randomly assigned to six dietary treatments with six replicates (cage) of 6 birds per cage. Dietary treatments consisted of a Control diet without antibiotics (C), and diets containing 11 mg/kg of "virginiamycin", 0.25% of yeast cell wall (YCW), 0.2% of a commercial product "Maxi-Gen Plus" containing processed yeast and nucleotides, 0.05% of nucleotides, or a diet containing 8% of distiller's dried grains with solubles (DDGS). On d 21 post-hatch blood samples were collected from 6 birds per treatment and serum sample were analyzed for antibody levels. After blood sampling, birds were injected intraperitoneally with 3 mg/kg of BW of lipopolysaccharide (LPS). The unchallenged group was fed the Control diet and injected with saline solution. Spleen samples were collected to measure the gene expression of toll-like receptors (TLR)2b, TLR4, and TLR21, macrophage mannose receptor (MMR), and cytokines including interleukin (IL)-12, IL-10, IL-4, IL-6, IL-18, and interferon (IFN)-γ. No significant difference in body weight gain, feed intake, and FCR were observed among treatments. Regarding humoral immunity, the diet supplemented with YCW increased serum immunoglobulin (Ig)A level compared with the antibiotic group; however, serum concentrations of IgG and IgM were not affected by dietary treatments. Relative gene expression of TLR2 and TLR4 was not affected by dietary treatments, whereas the expression of TLR21 and MRR was upregulated in diets containing YCW and DDGS. The diet supplemented with YCW increased the expression of all cytokines, and expression of IFN-γ was upregulated in the DDGS group. However, no significant difference was observed for cytokine gene expression in the antibiotic and nucleotide diets. In conclusion, supplementation of diet with YCW stimulated the systemic innate immune responses of broiler chickens following challenge with LPS. [ABSTRACT FROM AUTHOR]

Under commercial conditions and during production periods, pigs are generally exposed to a number of stressors that may have direct or indirect influence on their performance and general health. As a result, environmental stressors can influence the productivity as well as the onset and severity of infectious diseases. The aim of this study was to investigate the effects of crowding stress on performance and some immunological responses in piglets challenged with enterotoxigenic Escherichia coli (ETEC) K88+ and fed diets containing anti-ETEC probiotic strains derived from E. coli in a 3-wk trial. Ninety-six piglets at 20 ± 1 d of age were randomly assigned to 6 experimental treatments with 3 pigs/pen and 5 replicates/treatment and fed a basal mash diet. To model stress, piglets were housed in 2 kinds of pens: standard floor allowance (SFA; no stress) and half standard floor allowance (HSFA; stressed). The treatments were 1) control pigs with SFA, 2) control pigs with HSFA, 3) pigs with SFA and challenged with ETEC (ESFA), 4) pigs with HSFA and challenged with ETEC (EHSFA), 5) pigs with ESFA and supplemented with a cocktail of E. coli probiotics (PSFA), and 6) pigs with EHSFA and supplemented with a cocktail of E. coli probiotics (PHSFA). After 4 d acclimatization, crowding stress was started on d 5, E. coli probiotics were given daily from d 5, and ETEC challenge was administered on d 8 and 10. Body weight and feed disappearance were determined weekly and severity of diarrhea was characterized daily. Blood samples were collected for biochemistry and inflammatory analysis and pigs were euthanized to obtain digesta for bacterial enumeration. The ADG and ADFI decreased (P < 0.05) in stressed and ETEC-challenged pigs 5 d after ETEC challenge. The ETEC population was higher in both ileal and colon digesta of stressed pigs whereas ETEC-challenged and probiotic-fed pigs had higher fecal consistency scores 96 h and 7 d after ETEC challenge. Stressed pigs had higher (P < 0.001) levels of neutrophils, neutrophihlymphocyte ratio, albumimglobulin ratio, fibrinogen, total proteimfibrinogen ratio, and Cortisol. Generally, the levels of Cortisol, interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) differed over time among some treatment groups. In conclusion, reduced space allowance stress and ETEC challenge depressed certain aspects of performance and immunological responses and increased E. coli counts; however, probiotics feeding had little positive effects on these measures suggesting further research. [ABSTRACT FROM AUTHOR]

The turnover of intestinal epithelial cells is a dynamic process that includes adequate cell pro-liferation and maturation in the presence of microbiota and migration and seeding of immune cells in early gut development in chickens. We studied the effect of yeast-derived macromolecules (YDM) on performance, gut health, and immune system gene expression in the in-testine of broiler chickens. One thousand eighty 1-d-old birds, with 60 birds per pen and 6 pens per treatment, were randomly assigned to 3 treatment diets; a diet con-taining monensin (control), control diet supplemented with bacitracin methylene disalycylate (BMD), and BMD diet supplemented with YDM. Feed intake, BW, mortality, ileum histomorphology, and gene expression of Toll-like receptors (TLR2b, TLR4, and TLR21), cy-tokines [interferon (IFN)-γ, IFN-β, IL-12p35, IL-1β, IL-6, IL-10, IL-8, IL-2, IL-4, and transforming growth factor (TGF)-4β], and cluster of differentiation (CD)40 in the ileum, cecal tonsil, bursa of Fabricius, and spleen were assessed. No significant overall difference in performance in terms of feed intake, BW gain, and G:F was observed among treatments (P > 0.05). The YDM diet resulted in significantly higher villi height and villi heightxrypt depth ratio compared with BMD and con-trol diets (P < 0.05). A significantly lower mortality was observed in the YDM treatment compared with both control and BMD treatments. Compared with the control, gene expression analysis in YDM treat-ment showed no major change in response in the ileum, whereas higher CD40, IFN-β, IL-β, IL-6, TGF-β4, IL-2, and IL-4 in the cecal tonsil; TLR2b, TLR4, TLR21, and TGF-β4 in the bursa of Fabricius; and TLR4, IL-12p35, IFN-γ, TGF-β4, and IL-4 in the spleen was ob-served (P < 0.05). In conclusion, supplementation of YDM supports pro- and anti-inflammatory cytokine production via T helper type 1 and 2 (Thl and Th2) cell-associated pathways both locally and systemically with a stronger additive effect in the cecal tonsil in the presence of BMD in the diet of chickens. [ABSTRACT FROM AUTHOR]

The combined effects of probiotics {Lactobacillus acidophilus, Lactobacillus casei, Streptococcus faecium, and Saccharomyces cerevi-siae) and organic acids (sorbic and citric acid) on intestinal morphology and expression of immune-related genes were investigated. One-day-old chicks were randomly assigned to 1 of 3 groups: birds not receiving probiotic or organic acids (control; T1), or birds receiving an oral combination (1 g/L in water) of 108 CFU/g of each of the aforementioned probiotics and organic acids (1% sorbic acid and 0.2% citric acid) for 7 (T2) or 14 d (T3). Each group was divided into 5 replicate pens of 20 birds each, and 5 birds from each group (1 from each pen) were killed on d 11 and 22. Intestinal sections were collected for histological assessment, and reverse transcriptase-PCR analysis was used to assess defensin and cathelicidins expression. Quantitative real-time PCR was used to assess toll-like receptors (T1R) and cytokine expression. Duodenal villus height was greater in T2 and T3 at d 11 (P≤ 0.036) and 22 (P ≤ 0.015) compared with T1. At d 11, duodenal goblet cell/unit area was less in T3, whereas it was greater in T2 compared with T1 in the jejunum (P = 0.009). Ileal goblet cell/unit area was greater in T3 at d 22 compared with T1 (P < 0.001). Avian beta-defensin-3 was expressed in all tissues except the bursa of T3 birds at d 11, and T1R-2 was down regulated in the cecal tonsil of birds in T2 and T3 at d 11 compared with T1 (P = 0.020 and 0.003, respectively). Expression of IL-12p35 in the ileum at d 11 was down regulated in T2 and T3 compared with T1 (P = 0.030 and 0.012, respectively). Reduced expression of INF-y was observed in the ileum in T3 compared with T1 at d 11 (P = 0.047). Ileal IL-6 and IL-10 and cecal tonsil interferon-gamma INF-&ggr; expressions were greater T2 at d 22 (P < 0.047) than T1. In conclusion, supplementation of combined probiotics and organic acids resulted in inconsistent gut morphology associated responses, and avian beta-defensins and cathelicidins expression were not associated with combined probiotics and organic acids supplementation. Birds supplemented with combined probiotics and organic acids for 7 d showing similar responses in T1R-2, IL-12p35, and IFN-y compared with those supplemented for 14 d indicates that shorter periods of supplementation might be enough to elicit beneficial responses. [ABSTRACT FROM AUTHOR]

Different aspects of folic acid (FA) transport in the intestine of the laying hen have been characterized. Less is known about the adaptive response of this process to a dietary challenge. To this end, a study was conducted to evaluate the effect of increased dietary FA supplementation on the rate of intestinal FA transport and the expression of the intestinal folate transporter genes, the proton-coupled folate transporters (PCFT) and the reduced folate carrier (RFC), in the laying hen. Twenty-four Shaver White hens at 34 wk of age were randomly assigned to receive 1 of 3 dietary treatments: 1) basal diet with no supplemental folate (n = 8), 2) basal diet + 10 mg/kg of crystalline FA (n = 8), and 3) basal diet + 100 mg/kg of crystalline FA (n = 8). A completely randomized design with 3 dietary treatments was used. Production performance was not affected by FA supplementation; however, egg and plasma folate concentrations increased (P < 0.001), whereas plasma homocysteine concentrations decreased (P < 0.011) in birds fed 10 or 100 mg of FA/kg of diet, relative to controls. Mucosal to serosal uptake of FA in the duodenum was decreased (P < 0.002), but the mRNA levels of the duodenal PCFT and RFC genes were not affected when birds were fed 10 or 100 mg of FA/kg of diet. In the jejunum, the mucosal to serosal uptake of FA, as well as the mRNA levels of the PCFT and RFC genes, were not influenced by increased FA supplementation. Overall, increased dietary levels of FA resulted in decreased transport of FA in the duodenum but not in the jejunum of laying hens. This decrease was not associated with decreased mRNA expression of the duodenal PCFT and RFC genes. Therefore, a posttranscriptional or translational adaptation of the intestinal folate transporters may be involved in the much lower transport of FA in the duodenum of laying hens during increased dietary supplementation of FA. [ABSTRACT FROM AUTHOR]

Absorption at the level of the intestine is likely a primary regulatory mechanism for the deposition of dietary supplemented folic acid into the chicken egg. Therefore, factors affecting the intestinal transport of folic acid in the laying hen may influence the level of egg folate concentrations. To this end, a series of experiments using intestinal everted sacs were conducted to characterize intestinal folic acid absorption processes in laying hens. Effects of naturally occurring folate derivatives (5-methyl and 10-formyltetrahydrofolate) as well as heme on folic acid absorption were also investigated. Folic acid absorption was measured based on the rate of uptake of 3H-labeled folic acid in the everted sac from various segments of the small and large intestines. Folic acid concentration, incubation length, and pH condition were optimized before the perforrnance of uptake experiments. The distribution profile of folic acid transport along the intestine was highest in the upper half of the small intestine. Maximum uptake rate (nrnol.100 g tissue'.rnin1) was observed in the duodenum (20.6 ± 1.9) and jejunurn (22.3 ± 2.0) and decreased significantly in the ileum (15.3 ± 1.1) and cecum (9.3 ± 0.9). Transport increased proportionately (P < 0.05) between 0.000 1 and 0.1 μM folic acid. Above 0.1 μM, the slope of the regression line was not significantly different from zero (P < 0.137). Folic acid uptake in the jejunum showed a maximum rate of transport at pH 6.0, but was lowest at pH 7.5. The presence of 5-methyl and 10-formyltetrahydrofolate as well as heme impeded folic acid uptake, reducing intestinal folic acid absorption when added at concentrations ranging from 0 to 100 μM. Overall, these data indicated the presence of a folic acid transport system in the entire intestine of the laying hen. Uptake of folic acid in the cecum raises the likelihood of absorption of bacterial-derived folate. [ABSTRACT FROM AUTHOR]

The objectives of the current study included the characterization of the temporal changes in indices of sulphur amino acid metabolism in piglets in response to vitamin B6 deficiency and repletion with graded levels of pyridoxine hydrochloride. In Experiment 1, 12 piglets (average initial weight = 55.3 kg; n = 56 per group) were fed a semi-purified diet containing either 0 (deficiency group) or 3 mg (control group) pyridoxine · HCl/kg diet, using a pair-feeding design, for 6 weeks. Piglets consuming vitamin B6-deficient diets exhibited decreased average daily gains on the 4th week and feed conversion efficiency from the 4th week until the end of the trial ( P < 0.05). Plasma pyridoxal-50 -phosphate (PLP), in pigs consuming vitamin B6-deficient diets, was significantly lower than controls throughout the experiment ( P < 0.01), reaching a nadir of 14% of the control animals' value by the end of the trial. Indices of sulphur amino acid metabolism, including activities of hepatic cystathionine-b-synthase (CBS), cystathionine-g-lyase (CGL) and serine hydroxymethyltransferase, as well as hepatic-free cysteine concentrations were markedly decreased after 6 weeks of B6 deficiency ( P < 0.05). Total hepatic mRNA expressions for CBS and CGL were not affected. Concurrently, hepatic-free homocysteine concentrations increased by more than eight-fold ( P < 0.01) at the end of the trial. An examination of plasma total homocysteine and cysteine concentrations revealed significant ( P < 0.05) differences between treatments, with evidence of an abrupt shift in concentrations at 3 weeks post-initiation of dietary treatments (.25-fold increase in homocysteine; halving of cysteine values). At the end of Experiment 1, vitamin B6 deficiency significantly increased plasma methionine and serine levels, but decreased plasma glycine concentrations ( P,0.05). In Experiment 2, 20 pigs of 14 days old (initial BW = 55.0 kg) were subjected to a 4-week vitamin B6 depletion protocol, based on results obtained in Experiment 1. After the depletion period and assessment of baseline status (four pigs), remaining pigs were allocated to one of four dietary vitamin B6 repletion treatments: 0.75, 1.5, 2.25 and 3 mg/kg diet as pyridoxine · HCl (n54 per level) for 14 days. Significant dose-dependent increases in plasma PLP and cysteine, and decreases in homocysteine were observed, and these were sensitive to the duration of repletion. In conclusion, data from the current studies support the use of both plasma PLP and homocysteine as sensitive indices of vitamin B6 status in the pig. Additionally, the observed patterns of responses in vitamin B6-sensitive metabolites are supportive of an inclusion level of 2.25 mg/kg diet, as pyridoxine · HCl, in diets for young pigs. [ABSTRACT FROM AUTHOR]

This study investigated the response of piglets receiving a nu-cleotide-rich yeast extract (NRYE) without or with feed enzymes (ENZ) and antimicrobial growth promoters (AGP) on performance, blood cell profile, serum and ileum cytokines, and gut structure after an Escherichia coli lipopolysaccharide (LPS) challenge. Thirty-six pigs were allotted to 6 diets including: a non-challenged control (CON), LPS-challenged control (LPS), LPS + AGP, LPS + NRYE, LPS + ENZ, and LPS + ENZ + NRYE. On Day 7, pigs were bled and thereafter injected with LPS or sterile saline. Blood samples were collected at 6, 48, and 96 h post-challenge (hpc). After 96 hpc, pigs were euthanized to obtain duodenum, jejunum, and ileum tissues. Overall (d 1 to 11), NRYE supplementation tended to attenuate LPS-induced reduction in gain/feed ratio (P = 0.096), but LPS + AGP pigs had higher body weight (P = 0.04) and average daily gain (P = 0.03) than LPS + NYRE pigs. At 6 hpc, LPS + NRYE and LPS + ENZ + NRYE pigs had lower (P < 0.05) plasma urea N (PUN) and higher (P < 0.05) platelet count than LPS pigs. At 96 hpc, LPS-induced villus atrophy in the jejunum and duodenum was attenuated (P < 0.05) in LPS + ENZ + NRYE pigs. At 6 hpc, serum TN-F-α and IL-10 concentrations were reduced (P < 0.05) and increased (P < 0.05), respectively, in pigs receiving AGP, NRYE, and ENZ + NRYE. At 96 hpc, compared with LPS pigs, ileal TNF-α expression was lowered in LPS + NRYE (P = 0.01) and LPS + ENZ + NRYE (P = 0.01) pigs, whereas ileal IL-βp and IL-10 expression was lower in all treatments (P < 0.05). In conclusion, LPS-challenged piglets fed diets supplemented with NRYE without or with enzymes expressed similar beneficial responses as those fed diets with AGP in terms of lowering PUN concentration, reducing duodenal and ileal villi atrophy, and downregulating serum and ileal proinflammatory cytokines. This suggests that supplementation of NRYE can promote the health of piglets during early weaning in antibiotic-free feeding regimens. [ABSTRACT FROM AUTHOR]

The reduced folate carrier (RFC) has been postulated to be a major entity for folate transport activity in humans and other mammals. However, there are limited reports of the importance of RFC in an avian system. In the current study, therefore, the molecular cloning and tissue distribution of RFC, as well as the effect of dietary folate supplementation on the expression of this transporter, were investigated in the chicken. Shaver White laying hens (n = 8 per diet) received 3 wheat-based diets containing the following: 1) no supplemental folate, 2) folic acid (10.00 mg/kg), or 3) 5-methyltetrahydrofolate (11.30 mg/kg) for 21 d. The mRNA expression levels were analyzed by quantitative real-time PCR. The results showed that the cloned partial RFC cDNA containing the full coding region from duodenum was 99% homologous to the reference gene available in GenBank. A broad expression profile of RFC transcripts was observed, with RFC mRNA detected in the brain, liver, kidney, spleen, lung, intestine, ovary, and testis, as well as other tissues. Realtime PCR analysis revealed that no significant differences (P > 0.05) due to diet were found in the mRNA levels of RFC in the duodenum and cecurn. However, compared with the basal diet, jejunal mRNA levels of RFC were decreased (P < 0.05) in hens fed with the 5-methyltetrahydrofolate diet, but the reduction did not reach significance (P = 0.077) in the hens fed the folic acid diet. Overall, the current study demonstrated that the RFC cDNA containing the full coding region was successfully cloned from the duodenum of laying hens. The wide tissue distribution of RFC transcripts is suggestive of an important role of RFC in the process of folate transport in the chicken. Moreover, dietary folate supplementation could downregulate the jejunal mRNA expression of RFC. Such findings will lay the foundation of future work involving the RFC in avian systems, including laying hens. [ABSTRACT FROM AUTHOR]

Dairy calves are colonized shortly after birth by multidrug resistant (MDR) bacteria, including Escherichia coli. The role of dairy colostrum fed to calves as a potential source of MDR bacteria resistance genes has not been investigated. This study determined the recovery rate of extended-spectrum cephalosporin-resistant (ESC-R) E. coli in colostrum from cows. The ESC-R E. coli isolates were further investigated to determine their phenotypic antimicrobial resistance pattern and the genes conferring ESC-R. Fresh colostrum was collected from 452 cows from 8 dairy herds in New Brunswick, Canada. The ESC-R E. coli was isolated from the colostrum by using the VACC agar, a selective media for extended-spectrum β-lactamase producing Enterobacteriaceae. Minimum inhibitory concentration was determined for all the suspected ESC-R E. coli isolates using a commercial gram-negative broth microdilution method. Two multiplex PCR were conducted on all the suspected ESC-R E. coli isolates to determine the presence of the blaCTX-M (groups 1, 2, 9, and 8/25) blaCMY-2, blaSHV, and blaTEM resistance genes. The ESC-R E. coli were detected in 20 (4.43%) of the colostrum samples. At least 1 ESC-R E. coli isolate was detected in 6 (75%) of the dairy herds. All ESC-R E. coli had MDR profiles based on minimum inhibitory concentration testing. No blaCTX-M groups genes were detected; however, the blaCMY-2 gene was detected in 9 or 20 (45%) and blaTEM was detected in 7 of 20 (35%) of the ESC-R E. coli. No ESC-R E. coli had both blaCMY-2 and blaTEM resistance genes. This is the first report of blaCMY-2 and blaTEM genes found in E. coli isolates cultured from dairy colostrum to our knowledge. [ABSTRACT FROM AUTHOR]

Three experiments were conducted to evaluate the effect of yeast-derived carbohydrates (YDC), and a blend of probiotics and YDC (synbiotic, SNB) on serum IgG concentration, maternal-derived antibody (MDA) decay, and specific antibody-mediated immune response in chick pullets following immunization with T-cell dependent antigens. A total of 300 day-old pullet chicks were randomly assigned to 3 dietary treatments including: a basal diet (Control), and diets containing YDC, and SNB (Lactobacillus acidophilus, L. casei, Streptococcus faecium, and Bacillus subtilis, and YDC). In experiment one, on d 1 and wk 3, 4, 5, and 6, blood samples were collected and serum were analyzed by ELISA for total IgG (Y), and MDA against Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV). The second experiment examined the specific antibody against infectious bronchitis virus (IBV) in pullet chicks following vaccination against IBV at d 1. Finally, in experiment 3, on d 21 and 28 posthatch, 10 birds per treatment were immunized intramuscularly with both sheep red blood cells (SRBC) and bovine serum albumin (BSA), and 11 after immunization serum samples were analyzed by hemagglutination assay for antibody response to SRBC, and by ELISA for serum IgM and IgG response to BSA. The results demonstrated that diet containing SNB increased serum IgG at wk 3 posthatch. However, the decay rate of MDA against NDV and IBDV were not affected by dietary treatments. Birds fed YDC showed higher specific antibody response against IBV in wk 4, while both diets containing YDC and SNB decreased antibody response to IBV in wk 6. In addition, specific antibody response against SRBC and BSA was not affected by diets. In conclusion, supplementation of diet with SNB improved humoral immunity by increasing IgG concentration in serum, and modulated the adaptive antibody-mediated immune response against IBV. [ABSTRACT FROM AUTHOR]

Tetracyclines, which represent one of the most commonly used antibiotics for poultry, are known to be deposited in bones, where they can remain, despite the observation of appropriate withdrawal times. The aim of the study was to determine the concentration of oxytretracycline (OTC) residues in the bone and muscle of chickens, following the oral administration of a commercially available liquid formulation, and to test their cytotoxic effects on an in vitro cell culture model. Seventy-two 1-day-old broiler chickens were randomly allotted into 2 groups (control and treated animals). OTC (40 mg/kg BW) was administered via drinking water during the 1 to 5 and 20 to 25 days of life periods. At the end of the trial, the birds were slaughtered and the OTC residues in the target tissues were measured by means of liquid chromatography (LC) - tandem mass spectrometry (MS/MS). Cytotoxicity was assessed by evaluating the pro-apoptotic effect of the bone residues on the K562 erythroleukemic line and on the peripheral blood mononuclear cells (PBMC). In all the animals, the OTC residues in the muscle were far below the established MRL of 100 µg/kg. The OTC levels in the bones of the treated animals were instead found in the parts per million (ppm) range. Cell cytotoxicity was assessed by evaluating the pro-apoptotic effect of OTC bone residues on the haematopoietic cell system. This in vitro system has revealed a significant pro-apoptotic effect on both the K562 cell line and PBMC cultures. This result suggests potential human and animal health risks due to the entry of tetracycline residues contained in the bones of treated livestock into the food-chain. This could be of concern, particularly for canine and feline diets, as meat, bone meal, and poultry by-products represent some of the main ingredients of pet foods, especially in the case of dry pet food. Further studies are needed to define the underlying mechanisms of cytotoxicity and to evaluate the in vivo toxicological implications due to the observed in vitro effects. [ABSTRACT FROM AUTHOR]

Direct-fed microbials (DFM) are used to improve livestock health and performance. The effects of 2 DFM products, a blend of 3 Bacillus strains (DFMB) and a Propionibacteriumspp. (DFMP), on broiler performance, nutrient utilization, and immune responses were investigated. Day-old (n = 120) male broilers were divided into 24 groups of 5 birds and fed 3 wheat-based diets in mash form (8 groups per diet) from d 1 to 22. The control diet was fed without or with 7.5 × 104 cfu/g of either DFMB or DFMP. From d 19 to 21 fecal samples were collected for determination of total tract apparent retention (TTAR) of nutrients and AMEn. On d 21, feed intake and BW were determined. On d 22, 5 birds per treatment were killed by cervical dislocation to collect jejunal and ileal contents for determination of digesta viscosity and apparent ileal digestibility (AID) of nutrients, respectively, and ileum, cecal tonsil, and spleen tissues for Toll-like receptors (TLR) and cytokine expressions. Compared with the control, DFM did not affect BW gain and feed intake but DFMP reduced G:F (P < 0.01). Compared with the control (2,875 kcal/kg), birds fed on DFMB and DFMP had higher AMEn (2,979 and 2,916 kcal/kg, respectively; P < 0.05), whereas both DFM reduced the AID of DM (P < 0.001) and CP (P < 0.01). Furthermore, DFMP reduced TTAR of NDF (29.0 vs. 18.4%; P < 0.001), whereas both DFM increased TTAR of DM and fat (P < 0.001). Supplementing DFMP downregulated ileal expression of TLR-2b, IL-2, IL-4, IL-6, IL-10, and IL-13, whereas DFMB downregulated TLR-2b, IL-2, IL-4, and IL-6 in all 3 tissues, IL-10 in the spleen, and upregulated IL-13 in the spleen. In conclusion, the DFM did not improve performance but increased the AMEn of diet by possibly increasing DM and fat retention. Overall, both DFM showed an antiinflammatory effect in the ileum, but DFMB had more effects on local and systemic immunity than DFMP. [ABSTRACT FROM PUBLISHER]

The present study was conducted to investigate the effects of dietary folic acid (FA) supplementation on performance, serum biochemical indices, and mRNA abundance of intestinal folate transporters in young and older laying hens after acute lipopolysaccharide (LPS) challenge. Two experiments were conducted separately involving 48 Shaver White young laying hens (24 wk of age) in experiment 1 and 48 Shaver White older laying hens (58 wk of age) in experiment 2. Birds were fed 2 diets in a complete randomized design. The diets were wheat-soybean meal based, with or without supplemental 4 mg of FA/kg of diet. Birds were fed for 8 wk, during which time feed consumption and egg production were monitored. At the end of each feeding experiment, 6 hens from each dietary treatment were injected intravenously with 8 mg/kg of BW of either Escherichia coli LPS or sterile saline. Four hours after injection, blood and intestinal samples were collected for further analysis. Compared with the control, dietary FA supplementation increased egg weight and egg mass and decreased serum glucose levels in the young laying hens, and reduced serum uric acid in the older laying hens (P < 0.05). Relative to saline injection, plasma homocysteine, serum calcium, and phosphorus levels were found to be lower in both young and older laying hens after LPS challenge (P < 0.05). Other serum biochemical variables and the mRNA expression of 2 folate transport genes in the small and large intestine were differentially affected by LPS challenge, and some of those responses varied with the age of the birds. Additionally, interactions between diet and LPS challenge were specifically found in the older laying hens. In summary, in addition to improving production performance, there were effects of dietary FA supplementation and its interaction with LPS challenge on biochemical constituents, and age played a role in the development of responses to diet and bacterial LPS infections. [ABSTRACT FROM AUTHOR]

Folic acid plays a key role in nucleic acids and protein synthesis, and has been associated with anti-inflammatory effects in LPS-induced infections. To this end, a study was conducted to investigate the effects of dietary folic acid (FA) supplementation in older laying hens (58 to 66 wk of age) challenged with Escherichia coli lipopolysaccharide (LPS). A total of 24 Shaver White laying hens at 58 wk were fed 2 diets. The diets were wheat-soybean-based, with either 0 or 4 mg of supplemental FA per kg of diet. After 8 wk of feeding and at 66 wk, the hens were injected intravenously with 8 mg of LPS or saline per kg of BW. Four hours after injection, blood was collected and hens were euthanized to obtain spleen and cecal tonsils. The T cell subsets in the blood and the spleen (CD4+ and CD8-I-), to-tal IgG, and biochemical constituents (total protein, albumin, globulin, and fibrinogen) were not influenced (P > 0.05) by dietary FA supplementation. However, LPS injection decreased (P < 0.05) biochemical con-stituents, CD4+, and CD8+ cells in the blood, whereas CD4+:CD8+ ratio and total IgG increased (P < 0.05), and fibrinogen was not influenced. Gene expression in the spleen and cecal tonsils was not influenced by di-etary FA supplementation except a diet x challenge interaction for interleukin (IL)-8 in the spleen; IL-8 de-creased in FA-fed hens that were treated with LPS. Also, FA supplementation decreased the expression of IL-8 in cecal tonsils. Relative to saline-injected hens, expression of IL-lß, interferon-γ, and IL-10 increased in the LPS-injected hens in the spleen and cecal tonsils, IL-8 increased in LPS-injected hens only in the cecal tonsils, whereas Toll-like receptor 4, IL-4, IL-17, and IL-18 increased in the LPS-injected hens only in the spleen; however, LPS decreased expression of IL-13 in the cecal tonsils. In conclusion, FA did not affect in-flammatory responses in older laying hens; more studies are required to investigate possible protective effects of FA in laying hens. [ABSTRACT FROM AUTHOR]

We investigated the effects of dietary fo-lic acid (FA) supplementation on immunological pa-rameters in young laying hens under acute conditions of lipopolysaccharide (LPS) challenge. Twenty-four Shaver White laying hens at 24 wk were fed 2 diets in a completely randomized design. The diets were wheat-soybean based, with either 0 or 4 mg of supple-mental FA per kilogram of diet. At 32 wk of age, 6 hens from each dietary treatment were injected intra-venously with 8 mg/kg of BW of either LPS or saline. Four hours after injection, blood was collected and the hens were euthanized to obtain spleen and cecal tonsils. Heterophil:lymphocyte ratio, CD3+, CD4+, CD8+ T cells, and CD4+:CD8+ cells in the blood and spleen were not affected by dietary FA. Relative to saline-injected hens, LPS-injected hens had fewer (P < 0.05) CD3+, CD4+, CD8+, and CD4+:CD8+ cells in the blood, and no difference was found in the spleen. Total protein, albumin, and globulin were found to be higher (P < 0.05) in FA-supplemented hens compared with the control. However, total protein, albumin, and globulins decreased (P < 0.05) in the LPS-injected hens com-pared with the saline control. Expression of interleukin (IL)-1(3 in cecal tonsils decreased (P < 0.05) in FA-supplemented hens, but no dietary influence was found on the expression of other genes in both the spleen and cecal tonsils. Lipopolysaccharide upregulated (P < 0.05) expression of IL-10 and interferon (IFN)-7 in the spleen, and IL-1/3, IL-10, and IFN-7 in the cecal ton-sils, whereas the expression of Toll-like receptor (TLR)-4 and IL-8 was not influenced by LPS in the spleen and cecal tonsils. There was a diet x challenge interaction for total IgG, and cytokines IL-1/3 and IL-18 in the spleen as well as IL-18 in the cecal tonsils. In con-clusion, there were few interactions of dietary FA and LPS; however, FA increased biochemical constituents, enhanced generation of total IgG, as well as exhibiting pleoitropic effects in inflammatory responses. [ABSTRACT FROM AUTHOR]

A study was conducted to assess the effect of yeast-derived carbohydrates (YDC) on performance and innate immune responses of broiler chickens. In total, 1,080 one-day-old birds were randomly assigned to one of 3 dietary treatments (n = 360): a standard broiler diet containing monensin (control), control + bacitracin methylene disalycylate (BMD), and YDC treatment (control + YDC at 0.02%, 0.01%, and 0.005% for starter, grower, and finisher, respectively). Weekly BW, feed intake (FI), and feed conversion ratio (FCR) were recorded. Immune organ weights, gut morphology, gene expression, heterophil¡lymphocyte (H:L), and serum IgG were determined at d 42. No significant difference in FCR, FI, and mortality was observed among treatments. However, BW gain in starter phase was higher in control and YDC treatments compared with BMD treatment. Ileal villi height, crypt depth, and their ratio were not significantly different among treatments, whereas villi width was lower in control and YDC treatments compared with BMD treatment. The number of goblet cells per unit area in the ileum was lower in BMD treatment compared with control and YDC treatments. Expression of TLR2b and IL-6 in the ileum and cecal tonsils was not significantly different among treatments (P > 0.05). Expression of TLR4 was downregulated in YDC treatment compared with control in the ileum. Expression of IL-12p35 and IFN-γ were downregulated in the YDC treatment only in the cecal tonsils. Compared with the control, the expression of IL-10 in both the ileum and the cecal tonsils was downregulated in YDC treatment. Serum IgG and H:L ratio were lower and higher, respectively, in the YDC treatment compared with control and BMD treatments. In conclusion, dietary inclusion of YDC affected intestinal cytokines antiinflammatory profile on a gut location associated immune pathways manner, suggesting different immune pathways that require further studies in this field. [ABSTRACT FROM AUTHOR]

Necrotic enteritis (NE) caused by Cbstridium perfringens is a reemerging disease of economic importance in areas of the world where antibiotic growth promoters have been banned. The effect of mannan-oligosaccharide (MOS) supplementation in organic diets of broilers challenged with C. perfringens on performance, gut morphology, and innate immunity was investigated. Three hundred Ross-308 broilers were fed antibiotic-free certified organic starter and grower diets. On d 14, birds were orally challenged with 1 mL of C. perfringens culture at 3 x 1010 cfu/bird. Treatments consisted of a control no-challenge (CO; 0 g/kg of MOS in the basal diet), control challenge (COC, 0 g/kg of MOS in the basal diet), and MOS challenge (2 g/kg of MOS in the basal diet). Challenge of birds resulted in decreased feed intake and BW gain (P = 0.048 and P = 0.026, respectively). Even though supplementation of diet with MOS improved feed intake (P = 0.985), BW gain and G:F were not improved compared with those of the CO group (P = 0.026 and P = <0.001, respectively). There was no significant difference among treatments in jejunal and ileal villus height, crypt depth, and goblet cells/mm2 (P> 0.05). Quantitative real-time PCR showed that, in the ileum, the MOS diet resulted in an upregulation of toll-like receptor (TLR)2b, TLR~j, interleukin (IL)-12p35, and interferon (IFN)-'y compared with CO (P = 0.003, P = 0.018, and P = 0.024, respectively). In the cecal tonsil, challenging birds with C. perfringens resulted in an upregulation of TLR2b compared with CO (P = 0.036), and MOS resulted in an upregulation of TLR~j (P = 0.018). In conclusion, feeding a MOS-supplemented diet to C. perfringens-challenged broiler chickens did not improve performance and gut morphology-associated responses. However, MOS was capable of altering TLR and cytokine profiles, where dual TLR52 and TLR~ pathways were associated with MOS supplementation with subsequent upregulation of ileal IL-12p35 and IFN-'y, implying that MOS supplementation in C. perfringens-challenged chickens supports a proinflammatory effect via T-helper cell-i associated pathways. [ABSTRACT FROM AUTHOR]

The effects of a grain-based subacute ruminal acidosis (SARA) challenge (GBSC) and an alfalfa-pellet SARA challenge (APSC) on fermentation and endotoxins in the rumen and in the cecum, as well as on endotoxins in peripheral blood, were determined. Six nonlactating Holstein cows with cannulas in the rumen and cecum were used in the study. A 3 x 3 Latin square arrangement of treatments with 4-wk experimental periods was adopted. During the first 3 wk of each experimental period, all cows received a diet containing 70% forages [dry matter (DM) basis]. In wk 4 of each period, cows received 1 of the following 3 diets: the 70% forage diet fed during wk 1 to 3 (control), a diet in which 34% of the dietary DM was replaced with grain pellets made of 50% ground wheat and 50% ground barely (GBSC), or a diet in which 37% of dietary DM was replaced with pellets of ground alfalfa (APSC). Rumen pH was monitored continuously using indwelling pH probes, and rumen fluid, blood, cecal digesta, and fecal grab samples were collected immediately before feed delivery at 0900 h and at 6 h after feed delivery on d 3 and 5 of wk 4. The time for which rumen pH was below 5.6 was 56.4, 225.2, and 298.8 min/d for the control, APSC, and GBSC treatments, respectively. Compared with the control, SARA challenges resulted in similar reductions in cecal digesta pH, which were 7.07, 6.86, and 6.79 for the control, APSC, and GBSC treatments, respectively. Compared with the control, only GBSC increased starch content in cecal digesta, which averaged 2.8, 2.6, and 7.4% of DM for the control, APSC, and GBSC, respectively. Free lipopolysaccharide endotoxin (LPS) concentration in rumen fluid increased from 10,405 endotoxin units (EU)/mL in the control treatment to 30,715 and 168,391 EU/mL in APSC and GBSC, respectively. Additionally, GBSC increased the LPS concentration from 16,508 to 118,522 EU/g in wet cecal digesta, and from 12,832 to 93,154 EU/g in wet feces. The APSC treatment did not affect LPS concentrations in cecal digesta and feces. All concentrations of LPS in blood plasma were below the detection limit of >0.05 EU/mL of the technique used. Despite the absence of LPS in blood, only GBSC increased the concentration of LPS-binding protein in blood plasma, which averaged, 8.9, 9.5, and 12.1 mg/L for the control, APSC, and GBSC treatments, respectively. This suggests that GBSC caused translocation of LPS from the digestive tract but that LPS was detoxified before entering the peripheral blood circulation. The higher LPS concentration in cecal digesta in the GBSC compared with the APSC suggests a higher risk of LPS translocation in the large intestine in GBSC than in APSC. [ABSTRACT FROM AUTHOR]

It has been hypothesized that zinc (Zn) levels beyond those that are nutritionally required may favor the utilization of dietary lysine, and consequently reduce the level of its inclusion into the diet. Therefore, the possible effects of interaction between chelated Zn and the level of lysin& (Lys) on egg production and egg quality of laying hens were evaluated. In total, 720 ISA Brown layer hens aged 24 to 36 wk (early phase) and 48 to 60 wk (late phase) were allotted in a completely ran- domized factorial design that used 3 Zn and 5 Lys lev- els (6 replications, 8 birds/replication). All birds aged 37 to 47 wk (between early and late phases) were fed a standard diet and maintained under the same experi- mental design. The Zn levels used were 137, 309, and 655 mg/kg; and the Lys levels were 0.560, 0.612, 0.677, 0.749, and 0.851%. The optimal levels, of Lys digest- ibility were based on laboratory analyses with regard to the weighted average relation~hip between 83.5% di- gestibility and the total Lys from principal ingredients. There was no effect of interaction found between the dietary levels of Zn and Lys for most of the variables studied; however, each had an independent effect on the variables. An increase in Zn from 137 to 655 mg/kg had no significant effect (F> 0.05) on the performance of hens in both phases; however, it showed a significant effect on egg quality (P < 0.01), principally on mineral composition. Increased Zn resulted in decreased shell weight, percentage of ash, yolk ash deposition, and to- tal ash deposition. On the other hand, an increase in Lys from 0.560 to 0.851% significantly affected (P < 0.002) several performance parameters and the chemi- cal composition of the eggs, including feed intake, feed conversion efficiency, BW gain, egg weight, and produc- tion. In conclusion, there was no interaction found be- tween Zn and Lys, but higher dietary levels of chelated Zn reduced bird performance and egg quality param- eters, whereas higher Lys levels could be beneficial to bird performance and egg quality. [ABSTRACT FROM AUTHOR]

The conversion of folic acid (FA) to the biologically active 5-methyltetrahydrofolate (5-MTHF) is necessary for the deposition of folate in the egg. A study was conducted to compare egg folate concentrations, indices of folate status, and activities of folatedependent enzymes in response to equimolar intake of either FA or 5-MTHF in laying hens. Forty-eight laying hens, 24 wk of age, from 2 different strains (Shaver White and Shaver Brown) were randomly assigned to receive 1 of 3 (n = 8 per strain) dietary treatments: 1) basal diet with no supplemental folate, 2) basal diet + 10 mg/kg of FA, or 3) basal diet + 11.3 mg/kg of 5-MTHF for 3 wk. A completely randomized design with 3 dietary treatments and 2 laying hen strains in a 3 x 2 factorial arrangement was used. Data were subjected to ANOVA, using the PROC GLM procedure of SAS. Plasma homocysteine, serum, and egg folate concentrations; hepatic senile hydroxymethyltransferase; and methionine synthase activity were affected by dietary folate supplementation but, not by its form (FA and 5-MTHF). Relative to control hens, plasma homocysteine was decreased (P < 0.05) by 14.2%, whereas serum and egg folate were increased (P < 0.05) by 78.3 and 61.8%, respectively, in hens consuming either folate compound. Hepatic serine hydroxymethyltransferase and methionine synthase activity were increased and decreased (P < 0.05), respectively, in folate-fed birds compared with control-fed birds. Hepatic dihydrofolate reductase was influenced by both the addition and form of dietary folate, being higher (P < 0.05) in FA-fed birds than in 5-MTHF and control-fed birds. Feed efficiency was improved (P < 0.05) in 5-MTHF-fed birds relative to FA-fed birds. Strain of hen influenced serum folate and plasma homocysteine concentrations hut riot other indices of folate metabolism. Overall, FA and 5-MTHF have equivalent effects in enhancing egg folate concentrations and improving folate status in laying hens. Also, supplementation and form of folate may modulate the activity of folate-dependent enzymes. [ABSTRACT FROM AUTHOR]

Concerns regarding the welfare of laying hens raised in battery cages have led to the development of enriched cages that allow hens to perform natural behaviors including nesting, roosting, and scratching. This study was conducted to compare indices of production and welfare in birds housed in 2 different caging systems. Shaver White hens were housed from 21 to 61 wk in either conventional battery cages (n = 500; 10 cages; 5 hens/cage; floor space = 561.9 cm²/hen) or enriched cages (n = 480; 2 cages; 24 hens/cage; floor space = 642.6 cm²/hen) and were replicated 10 times. Enriched cages provided hens with a curtained nesting area, scratch pad, and perches. Production parameters and egg quality measures were recorded throughout the experiment. Plumage condition was evaluated at 37 and 61 wk. Bone quality traits and immunological response parameters were measured at 61 wk, and 59 and 61 wk, respectively. Hen-day egg production, feed consumption, egg weight, and percentage of cumulative mortality of laying hens were not affected by the cage designs. Specific gravity and the percentage of cracked and soft-shelled eggs were also similar between the 2 housing systems. The incidence of dirty eggs was, however, significantly higher (P < 0.0001) in enriched cages than in conventional cages. Feather scores were similar between birds except for the wing region, which was higher (P < 0.05) for hens housed in conventional cages. Bone quality measures tended to be higher for hens housed in enriched cages compared with hens in conventional cages. However, the increase was significant only for bone mineral density. Immunological response parameters did not reveal statistically significant differences. Overall, laying performance, exterior egg quality measures, plumage condition, and immunological response parameters appear to be similar for hens housed in the 2 cage systems tested. Enrichment of laying hen cages resulted in better bone quality, which could have resulted from increased activity. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of subclinical mastitis (SCM) on calving-to-first-service interval (CFS), calving-to-conception interval (CC), and on the number of services per conception (S/C) in grazing Holstein and Normande cows. Primiparous (n=43) and multiparous (n=165) cows were selected from five dairy herds. Two composite milk samples were aseptically collected from each cow at drying-off, and then every week during the first postpartum month. One sample was used for somatic cell count (SCC), and the other one for bacteriological analysis. Cows were followed up to 300 d after calving. Non-parametric and parametric survival models, and negative binomial regression were used to assess the association between SCM, evaluated by SCC and milk culture, and reproductive indices. Staphylococcus aureus, CNS, and Streptococcus uberis were the most frequent isolated pathogens. Subclinical mastitis in the first month of lactation was not associated with CFS; however, the CC interval was longer in cows with SCM compared to healthy cows, the former also had a higher number of S/C., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Vitamin D requirement is estimated to be higher than recommended values for the first two weeks of a broiler chicken's life, and is heavily dependent on the concentrations of Ca and P in the diet. There are data indicating the beneficial effect of higher vitamin D levels on performance and overall health of the chickens. However, data on the role of higher vitamin D levels on the innate immune response of chickens are limited. Therefore, in the current study, we examined the effect of higher doses of vitamin D supplementation on the innate immune response in broiler chickens receiving optimal or calcium (Ca) and phosphorus (P) deficient diets. Three hundred Ross-308 male broiler chicks were randomly allocated into 60 cages with 5 birds per cage in a 3 × 2 factorial design with three levels of vitamin D and two levels of Ca/P with each experimental diet fed to 10 cages (10 replicates). Quantitative reverse transcription PCR (n = 5) was used to assess Toll-like receptor (TLR2b and 4), cytokine/chemokine (IL-12, IFN-γ, IL-10, IL-4, IL-13, IL-18, CxCLi2) and cathelicidin (CATH1, CATHB1, CATH3) transcription levels in peripheral blood mononuclear cells (PBMCs), spleen, and bursa of Fabricius. Vitamin D supplementation of the Ca and P deficient diet considerably augmented transcription of TLR2b, TLR4, CATH1, and CATHB1 and predominantly Th2 cytokines in spleen. Supplementation of the control diet with vitamin D downregulated TLR4 transcription, and dose-dependently increased CATH1, CATHB1, Th1, and Th2 cytokine transcription (Th2>Th1). All diets downregulated CATH3 transcription. In conclusion, vitamin D or its derivative 25-OH-D 3 both have a robust immunomodulatory property with a more favorable Th2 response, while at the same time enhancing observed Th2 cytokine responses under both optimal and lower Ca and P inclusion levels in the diets of broiler chickens., (© 2016 Poultry Science Association Inc.)

This study evaluated the effect of yeast-derived products on growth performance, gut lesion score, intestinal population of Clostridium perfringens, and local innate immunity of broiler chickens challenged with C. perfringens. One-day-old broiler chickens were randomly assigned to eight dietary treatments providing six replicate pens of 55 birds each per treatment. Dietary treatments consisted of Control diets without and with C. perfringens challenge, and diets containing bacitracin methylene disalicylate (BMD, 55 g/tonne), nucleotides (150 g/tonne), yeast cell wall (YCW, 300 g/tonne), and a commercial product Maxi-Gen Plus (1 kg/tonne) fed to chickens challenged with C. perfringens. Diets containing 10% distillers dried grains with solubles without and with C. perfringens challenge were also used. Birds were orally challenged with C. perfringens (10(8) colony-forming units (cfu)/bird) on day 14. On day 21, intestinal samples were collected for gene expression analysis. Pathogen challenge significantly (P < 0.05) impaired feed intake, body weight gain, and feed conversion ratio (FCR) shortly after the challenge (14-21 days). Increased C. perfringens counts and intestinal lesion scores were observed for challenged birds except the BMD-containing diet. Over the entire trial (1-35 days), no difference in growth performance was observed except the BMD diet which improved FCR over the Control, challenged group. Birds receiving nucleotides showed increased expression of toll-like receptors and cytokines interleukin (IL)-4 and IL-18 compared to the Control, challenged group. Expression of macrophage mannose receptor and IL-18 was upregulated in birds receiving YCW. Increased expression of cytokines and receptors involved in innate immunity in broilers receiving nucleotides and YCW suggests the immunomodulatory properties of these products under pathogen challenge conditions.

Prebiotics and synbiotics are considered to be among the most promising replacements for in-feed antibiotic growth promoters (AGP) in poultry feed. The current study was designed to study the effect of Bacitracin methylene disalicylate (BMD) (Control), yeast-derived carbohydrates (YDC), and a blend of YDC and probiotics [Lactobacillus acidophilus, Lactobacillus casei, Streptococcus faecium, Bacillus subtilis, and YDC] (SNB) in the performance and innate immune response of pullets. Feed intake and BW were measured on a weekly basis. At the end of the study (d 21), 10 birds/treatment were sacrificed by cervical dislocation and ileum, cecal tonsil, and spleen samples were collected for gene expression analysis. No significant difference (P > 0.05) in feed intake and G:F was observed among treatments. In the second and third wk age, higher BW gain was observed in SNB treatment compared to control and both control and YDC treatments, respectively. Expression of TLR2b was upregulated in YDC and SNB in the ileum, and in SNB in the spleen (P < 0.05). Expression of TLR4 was downregulated in SNB in the cecal tonsil. Expression of TLR21 was downregulated in YDC in the ileum, while it was upregulated in SNB in the spleen (P < 0.05). In the ileum, YDC resulted in downregulated IL-12p35, CxCLi2, and IL-13, and SNB resulted in upregulated IL-6, interferon (IFN)-γ, and IL-4 (P < 0.05). In the cecal tonsil, YDC resulted in upregulated IL-12p35, IL-2, IL-13 and IL-10, and SNB resulted in downregulated IL-2 and upregulated IL-13 and IL-10 (P < 0.05). In the spleen, YDC resulted in dowregulated IL-2 and CxCLi2, and SNB resulted in upregulated IL-6, IFN-γ, IL-4, and IL-10 (P < 0.05). In conclusion, no change in performance was observed. Innate immune response analysis showed SNB with a more potent effect compared to YDC where the former showed a balanced T-helper (Th)-1/Th-2 response locally and a more Th-2-dependent response systemically; SNB might provide a more beneficial immune modulation with maintaining immune homeostasis, which was observed in a strong IL-10 response., (© 2015 Poultry Science Association Inc.)

Background: The role of transforming growth factor-beta 1 (TGF-β₁) in the onset of bone marrow fibrosis has been confirmed in some animal models. To further understand the genetic expression of some myeloproliferative disorders affecting marrow stem cells, however, it is necessary to develop a specific and reliable procedure to deliver modified adenoviral vectors into the bone marrow cavity. The aim of this paper is to report a surgical technique designed to deliver an adenoviral vector-mediated gene expressing TGF-β₁ into the bone marrow of rat femurs., Methods: Forty-two Sprague-Dawley rats were used in the study. Rat femurs were exposed and the compact and trabecular bones at the proximal head removed. An intrabone marrow injection of a mutated TGF-β₁ adenoviral vector, a null adenoviral vector, or PBS was delivered into the bone. Three groups were accounted (n = 14 per group): fibrogenic and positive and negative controls. The quality of the surgical entrance was assessed by means of computerized tomography and histological changes were assessed by histochemistry. The concentration of TGF-β₁ in the bone marrow was determined by ELISA., Results: The surgical technique was conducted under ideal timing (approx. 10 min) and no surgical or postsurgical complications were observed. Computerized tomography revealed no changes in the bone tissue and a clean entrance was delimited through the bone to the bone marrow. HE and Masson's trichrome staining indicated highly fibrotic areas in the profibrotic group and bone marrow lavage reported a significantly higher concentration of TGF-β₁ (p < 0.05) in that same group., Conclusions: The present study confirmed that the proposed surgical technique is an effective method to deliver adenoviral vectors into the femoral bone marrow to investigate the physiopathology of bone marrow fibrosis in rats., (Copyright © 2013 S. Karger AG, Basel.)

Necrotic enteritis (NE) caused by Clostridium perfringens is a reemerging disease of economic importance in areas of the world where antibiotic growth promoters have been banned. The effect of mannan-oligosaccharide (MOS) supplementation in organic diets of broilers challenged with C. perfringens on performance, gut morphology, and innate immunity was investigated. Three hundred Ross-308 broilers were fed antibiotic-free certified organic starter and grower diets. On d 14, birds were orally challenged with 1 mL of C. perfringens culture at 3 × 10(10) cfu/bird. Treatments consisted of a control no-challenge (CO; 0 g/kg of MOS in the basal diet), control challenge (COC, 0 g/kg of MOS in the basal diet), and MOS challenge (2 g/kg of MOS in the basal diet). Challenge of birds resulted in decreased feed intake and BW gain (P = 0.048 and P = 0.026, respectively). Even though supplementation of diet with MOS improved feed intake (P = 0.985), BW gain and G:F were not improved compared with those of the CO group (P = 0.026 and P = <0.001, respectively). There was no significant difference among treatments in jejunal and ileal villus height, crypt depth, and goblet cells/mm(2) (P > 0.05). Quantitative real-time PCR showed that, in the ileum, the MOS diet resulted in an upregulation of toll-like receptor (TLR)2b, TLR4, interleukin (IL)-12p35, and interferon (IFN)-γ compared with CO (P = 0.003, P = 0.018, and P = 0.024, respectively). In the cecal tonsil, challenging birds with C. perfringens resulted in an upregulation of TLR2b compared with CO (P = 0.036), and MOS resulted in an upregulation of TLR4 (P = 0.018). In conclusion, feeding a MOS-supplemented diet to C. perfringens-challenged broiler chickens did not improve performance and gut morphology-associated responses. However, MOS was capable of altering TLR and cytokine profiles, where dual TLR2 and TLR4 pathways were associated with MOS supplementation with subsequent upregulation of ileal IL-12p35 and IFN-γ, implying that MOS supplementation in C. perfringens-challenged chickens supports a proinflammatory effect via T-helper cell-1 associated pathways.

The effects of a grain-based subacute ruminal acidosis (SARA) challenge (GBSC) and an alfalfa-pellet SARA challenge (APSC) on fermentation and endotoxins in the rumen and in the cecum, as well as on endotoxins in peripheral blood, were determined. Six nonlactating Holstein cows with cannulas in the rumen and cecum were used in the study. A 3×3 Latin square arrangement of treatments with 4-wk experimental periods was adopted. During the first 3 wk of each experimental period, all cows received a diet containing 70% forages [dry matter (DM) basis]. In wk 4 of each period, cows received 1 of the following 3 diets: the 70% forage diet fed during wk 1 to 3 (control), a diet in which 34% of the dietary DM was replaced with grain pellets made of 50% ground wheat and 50% ground barely (GBSC), or a diet in which 37% of dietary DM was replaced with pellets of ground alfalfa (APSC). Rumen pH was monitored continuously using indwelling pH probes, and rumen fluid, blood, cecal digesta, and fecal grab samples were collected immediately before feed delivery at 0900 h and at 6 h after feed delivery on d 3 and 5 of wk 4. The time for which rumen pH was below 5.6 was 56.4, 225.2, and 298.8 min/d for the control, APSC, and GBSC treatments, respectively. Compared with the control, SARA challenges resulted in similar reductions in cecal digesta pH, which were 7.07, 6.86, and 6.79 for the control, APSC, and GBSC treatments, respectively. Compared with the control, only GBSC increased starch content in cecal digesta, which averaged 2.8, 2.6, and 7.4% of DM for the control, APSC, and GBSC, respectively. Free lipopolysaccharide endotoxin (LPS) concentration in rumen fluid increased from 10,405 endotoxin units (EU)/mL in the control treatment to 30,715 and 168,391 EU/mL in APSC and GBSC, respectively. Additionally, GBSC increased the LPS concentration from 16,508 to 118,522 EU/g in wet cecal digesta, and from 12,832 to 93,154 EU/g in wet feces. The APSC treatment did not affect LPS concentrations in cecal digesta and feces. All concentrations of LPS in blood plasma were below the detection limit of >0.05 EU/mL of the technique used. Despite the absence of LPS in blood, only GBSC increased the concentration of LPS-binding protein in blood plasma, which averaged, 8.9, 9.5, and 12.1mg/L for the control, APSC, and GBSC treatments, respectively. This suggests that GBSC caused translocation of LPS from the digestive tract but that LPS was detoxified before entering the peripheral blood circulation. The higher LPS concentration in cecal digesta in the GBSC compared with the APSC suggests a higher risk of LPS translocation in the large intestine in GBSC than in APSC., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

1. The aim was to investigate the molecular characterisation and effects of dietary folate supplementation on mRNA concentrations of the proton-coupled folate transporter (PCFT) in chicken. 2. Twenty-four (n = 8/treatment) laying hens were allocated at random to one of three dietary treatments: a basal diet without supplemental folate, or the same diet with either 10·00 mg/kg folic acid or 11·30 (equimolar) mg/kg 5-methyltetrahydrofolate (5-MTHF) for 21 d. The PCFT cDNA was cloned using the TA cloning system, and mRNA concentrations were determined by real-time polymerase chain reaction (PCR). 3. The PCFT cDNA (1643 bp) was cloned from the duodenum and its transcripts were widely distributed in chicken tissues. Duodenal and caecal PCFT mRNA was not affected by dietary treatments; however, jejunal PCFT mRNA was significantly reduced in hens fed the 5-MTHF diet versus the basal diet. 4. A broad tissue distribution of PCFT transcripts is indicative of a vital role of PCFT in the folate transport in chicken. The expression of jejunal PCFT mRNA was down-regulated by dietary 5-MTHF supplementation. These findings contribute to an understanding of the development of folate transport and the PCFT system in avian systems involving laying hens.

The objectives of the current study included the characterization of the temporal changes in indices of sulphur amino acid metabolism in piglets in response to vitamin B6 deficiency and repletion with graded levels of pyridoxine hydrochloride. In Experiment 1, 12 piglets (average initial weight = 5.3 kg; n = 6 per group) were fed a semi-purified diet containing either 0 (deficiency group) or 3 mg (control group) pyridoxine·HCl/kg diet, using a pair-feeding design, for 6 weeks. Piglets consuming vitamin B6-deficient diets exhibited decreased average daily gains on the 4th week and feed conversion efficiency from the 4th week until the end of the trial (P < 0.05). Plasma pyridoxal-5'-phosphate (PLP), in pigs consuming vitamin B6-deficient diets, was significantly lower than controls throughout the experiment (P < 0.01), reaching a nadir of 14% of the control animals' value by the end of the trial. Indices of sulphur amino acid metabolism, including activities of hepatic cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CGL) and serine hydroxymethyltransferase, as well as hepatic-free cysteine concentrations were markedly decreased after 6 weeks of B6 deficiency (P < 0.05). Total hepatic mRNA expressions for CBS and CGL were not affected. Concurrently, hepatic-free homocysteine concentrations increased by more than eight-fold (P < 0.01) at the end of the trial. An examination of plasma total homocysteine and cysteine concentrations revealed significant (P < 0.05) differences between treatments, with evidence of an abrupt shift in concentrations at 3 weeks post-initiation of dietary treatments (>25-fold increase in homocysteine; halving of cysteine values). At the end of Experiment 1, vitamin B6 deficiency significantly increased plasma methionine and serine levels, but decreased plasma glycine concentrations (P < 0.05). In Experiment 2, 20 pigs of 14 days old (initial BW = 5.0 kg) were subjected to a 4-week vitamin B6 depletion protocol, based on results obtained in Experiment 1. After the depletion period and assessment of baseline status (four pigs), remaining pigs were allocated to one of four dietary vitamin B6 repletion treatments: 0.75, 1.5, 2.25 and 3 mg/kg diet as pyridoxine·HCl (n = 4 per level) for 14 days. Significant dose-dependent increases in plasma PLP and cysteine, and decreases in homocysteine were observed, and these were sensitive to the duration of repletion. In conclusion, data from the current studies support the use of both plasma PLP and homocysteine as sensitive indices of vitamin B6 status in the pig. Additionally, the observed patterns of responses in vitamin B6-sensitive metabolites are supportive of an inclusion level of 2.25 mg/kg diet, as pyridoxine·HCl, in diets for young pigs.