Effects of a postbiotic, with and without a saponin-based product, on turkey performance

ABSTRACT: Modern poultry production relies on an ability to prevent and mitigate challenges to bird health, while maintaining a productive bird. A number of different classes of biologics-based feed additives exist, and many have been tested individually for their impacts on poultry health and performance. Fewer studies have examined the combinations of different classes of products. In this study, we examined the use of a well-established postbiotic feed additive (Original XPC, Diamond V) on turkey performance, with and without the addition of a proprietary saponin-based feed additive. This was accomplished in an 18-wk pen trial utilizing 22 pen replicates per treatment across 3 treatments (control, postbiotic, and postbiotic plus saponin). Significant differences in body weight were identified at wk 12 and 15 of age, with the postbiotic plus saponin treatment group resulting in heavier birds at both timepoints. Significant differences in feed conversion ratio were observed from 0 to 18 wk of age, with the postbiotic alone having improved FCR compared with the control group. No significant differences were observed for livability or feed intake. This study demonstrates that a combination of a postbiotic plus saponin may exert additive effects on the growth of the turkey

Evaluation of gastrointestinal leakage in multiple enteric inflammation models in chickens

Enteric inflammation models can help researchers study methods to improve health and performance plus evaluate various growth promoters and dietary formulations targeted to improve performance in poultry. Oral administration of fluorescein isothiocyanate dextran (FITC-d; 3-5 kDa) and its pericellular mucosal epithelial leakage is an established marker to evaluate enteric inflammation in multiple species. The present study evaluated different methods to induce gut inflammation in poultry based on FITC-d leakage. Four independent experiments were completed with different inflammation treatment groups; serum FITC-d and/or retention of FITC-d in GI tract was/were determined. In experiment 1 (n=10 birds/treatment; broilers; processed at 14d), groups included control (CON), Dextran sodium sulfate (DSS; drinking water at 0.75%) and feed restriction (FRS; 24h before processing). Experiment 2 (n=14birds/treatment; leghorns; processed at 7d) included CON, DSS, FRS, and rye based diet (RBD). In experiments 3 and 4 (n=15 birds/treatment; broilers; processed at 7d), groups were CON, DSS, high fat diet (HFD), FRS, and RBD. In all experiments, FRS and RBD treatments showed significantly higher serum FITC-d levels compared to the respective CON. This indicates that FRS and RBD results in disruption of the intact barrier of the GIT, resulting in increased gut permeability. DSS and HFD groups showed elevation of serum FITC-d levels although the magnitude of difference from respective CON were inconsistent between experiments. FRS was the only treatment which consistently showed elevated retention of FITC-d in GIT in all experiments. The results from present studies showed that FRS and RBD, based on serum FITC-d levels, can be robust models to induce gut leakage in birds in different age and species/strains

Rye Affects Bacterial Translocation, Intestinal Viscosity, Microbiota Composition and Bone Mineralization in Turkey Poults

<div><p>Previously, we have reported that rye significantly increased both viscosity and <i>Clostridium perfringens</i> proliferation when compared with corn in an <i>in vitro</i> digestive model. Two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and bone mineralization, when compared with corn in turkey poults. In each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or a rye diet (<i>n</i> = 0 /group). At 10 d of age, in both experiments, 12 birds/group were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood and liver samples were collected to evaluate the passage of FITC-d and bacterial translocation (BT) respectively. Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of bone parameters. Broilers fed with a rye diet showed increased (<i>p</i><0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that turkey poults fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to turkey poults fed with corn. Turkey poults fed with rye also had significantly higher coliforms in duodenum and ileum but not in the ceca, whereas the total number of anaerobes increased only in duodenum. A significant reduction in bone strength and bone mineralization was observed in turkey poults fed with rye when compared with corn fed turkey poults. In conclusion, rye evoked mucosal damage in turkey poults that increased intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition and bone mineralization. Studies to evaluate dietary inclusion of selected Direct-Fed Microbial (DFM) candidates that produce exogenous enzymes in rye fed turkey poults are currently being evaluated.</p></div

Role of a Bacillus subtilis direct-fed microbial on digesta viscosity, bacterial translocation and bone mineralization in turkey poults fed with a rye-based diet

Rye contains high concentrations of non-starch polysaccharides (NSP), leading to reduced digestibility. Since poultry have little or no endogenous enzymes capable of hydrolyzing these NSP, exogenous carbohydrases as feed additives are used in an attempt to reduce the anti-nutritional effects of these polysaccharides. Previously, an in vitro study conducted in our laboratory showed that inclusion of certain Bacillus direct-fed microbial (DFM) candidates that produce exogenous phytase, lipase, protease, cellulase and xylanase in high NSP diets significantly reduced both digesta viscosity and Clostridium perfringens proliferation. In the present study, rye-based turkey starter diets with or without Bacillus-DFM were administered ad libitum to day-of-hatch turkey poults in two independent experiments. In both experiments, day-of-hatch turkey poults were randomly assigned to either a control diet (CON) or a DFM treated diet (n = 25 birds/group). At ten days–of-age, all turkey poults from experiments 1 and 2 were weighted and 12 turkey poults/group were randomly selected and humanely killed. Liver samples were aseptically collected to evaluate bacterial translocation, and intestinal digesta samples were individually collected to evaluate viscosity. Additionally, in experiment 2 both tibias were removed for assessment of bone parameters. In both experiments, the TRT group showed a reduction in the total number of coliforms in the liver as well as a reduced digesta viscosity when compared to the CON group (P<0.05). Turkey poults fed the Bacillus-DFM candidate had increased tibia diameter, breaking strength, ash content, calcium content, and phosphorus content when compared with CON turkey poults. In summary, turkey poults fed with a rye-based diet without DFM showed an increase in bacterial translocation and digesta viscosity, accompanied by a reduction in bone mineralization; however these adverse effects can be prevented by the inclusion of selected a Bacillus-DFM candid

A Fast and Inexpensive Protocol for Empirical Verification of Neutralizing Epitopes in Microbial Toxins and Enzymes

In vivo targeting of peptides to antigen-presenting cells by use of agonistic anti-CD40 monoclonal antibodies has been used successfully as an immune response enhancing strategy. When tested in chickens, the antibody-guided platform was capable of inducing specific IgG production within 1 week postimmunization. However, use of this method beyond its initial conception as a vaccine delivery tool has not been fully exploited. In this study, Clostridium perfringens alpha-toxin was used as a model microbial toxin for epitope mapping by using the antibody-guided immunization method to generate a panel of antibodies against specific, regions of the toxin in an attempt to identify crucial determinants on the toxin which, once bound, would hinder downstream toxicity. Alpha-toxin, which possesses both hemolytic and phospholipase C (PLC) enzymatic activities, has long been known to be one of the key destructive etiological agents of necrotic enteritis disease in poultry. Previous attempts to identify crucial antigenic determinants on the toxin mediating its enzymatic activities have been performed using expensive and labor-intensive site-directed mutagenesis techniques. To create a panel of antibodies, 23 short candidate alpha-toxin peptide regions were selected in silico using B-cell epitope prediction algorithms in the public domain and were custom synthesized to load onto the antibody-guided complex for immunization in birds for antisera production. Peptide-specific antibody responses were generated against all candidate neutralizing epitopes and used for in vitro toxin neutralization tests. Antisera against all 23 peptides were able to neutralize the toxin’s hemolytic activity, with neutralization titers ranging from 80 to 320, but none were effective in blocking PLC. The novel approach of antibody-guided immunization introduces a new, inexpensive method for polyclonal IgG production and de facto identification of neutralizing epitopes in microbial toxins and enzymes within 2 weeks from in silico analysis of a putative target sequence

Upstream Regulator Analysis of Wooden Breast Myopathy Proteomics in Commercial Broilers and Comparison to Feed Efficiency Proteomics in Pedigree Male Broilers

In an effort to understand the apparent trade-off between the continual push for growth performance and the recent emergence of muscle pathologies, shotgun proteomics was conducted on breast muscle obtained at ~8 weeks from commercial broilers with wooden breast (WB) myopathy and compared with that in pedigree male (PedM) broilers exhibiting high feed efficiency (FE). Comparison of the two proteomic datasets was facilitated using the overlay function of Ingenuity Pathway Analysis (IPA) (Qiagen, CA, USA). We focused on upstream regulator analysis and disease-function analysis that provides predictions of activation or inhibition of molecules based on (a) expression of downstream target molecules, (b) the IPA scientific citation database. Angiopoeitin 2 (ANGPT2) exhibited the highest predicted activation Z-score of all molecules in the WB dataset, suggesting that the proteomic landscape of WB myopathy would promote vascularization. Overlaying the FE proteomics data on the WB ANGPT2 upstream regulator network presented no commonality of protein expression and no prediction of ANGPT2 activation. Peroxisome proliferator coactivator 1 alpha (PGC1α) was predicted to be inhibited, suggesting that mitochondrial biogenesis was suppressed in WB. PGC1α was predicted to be activated in high FE pedigree male broilers. Whereas RICTOR (rapamycin independent companion of mammalian target of rapamycin) was predicted to be inhibited in both WB and FE datasets, the predictions were based on different downstream molecules. Other transcription factors predicted to be activated in WB muscle included epidermal growth factor (EGFR), X box binding protein (XBP1), transforming growth factor beta 1 (TGFB1) and nuclear factor (erythroid-derived 2)-like 2 (NFE2L2). Inhibitions of aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT) and estrogen related receptor gamma (ESRRG) were also predicted in the WB muscle. These findings indicate that there are considerable differences in upstream regulators based on downstream protein expression observed in WB myopathy and in high FE PedM broilers that may provide additional insight into the etiology of WB myopathy

High Molecular Weight Polymer Promotes Bone Health and Prevents Bone Loss Under Salmonella Challenge in Broiler Chickens

As a consequence of rapid growth, broiler chickens are more susceptible to infection as well as bone fractures that result in birds being culled. Intestinal infection/inflammation has been demonstrated to promote bone loss in mice and humans. Given this link, we hypothesize that therapeutics that target the gut can benefit bone health. To test this, we infected broiler chickens (7 days old) with Salmonella and treated the birds with or without MDY, a non-absorbable mucus supplement known to benefit intestinal health, from day 1–21 or from day 14–21. Chicken femoral trabecular and cortical bone parameters were analyzed by microcomputed tomography at 21 days. Birds infected with Salmonella displayed significant trabecular bone loss and bone microarchitecture abnormalities that were specific to the femoral neck region, a common site of fracture in chickens. Histological analyses of the chicken bone indicated an increase in osteoclast surface/bone surface in this area indicating that infection-induced bone resorption likely causes the bone loss. Of great interest, treatment with MDY effectively prevented broiler chicken bone loss and architectural changes when given chronically throughout the experiment or for only a week after infection. The latter suggests that MDY may not only prevent bone loss but reverse bone loss. MDY also increased cortical bone mineral density in Salmonella-treated chickens. Taken together, our studies demonstrate that Salmonella-induced bone loss in broiler chickens is prevented by oral MDY

Utilization of rye as energy source affects bacterial translocation, intestinal viscosity, microbiota composition and bone mineralization in broiler chickens

Two independent trials were conducted to evaluate the utilization of rye as energy source on bacterial translocation, intestinal viscosity, gut integrity, gut microbiota composition, and bone mineralization, when compared with a traditional cereal (corn) in broiler chickens. In each experiment, day-of-hatch, broiler chickens were randomly assigned to either a corn or a rye diet (n = 20 chickens/group). At 10d of age, in both experiments, 12 chickens/group were randomly selected, and given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood samples were collected to determine the passage of FITC-d. The liver was collected from each bird to evaluate bacterial translocation (BT). Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of bone parameters. Broilers fed with rye showed increased (p < 0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that chickens fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to chickens fed with corn. Chickens fed with rye also had significantly higher coliforms in duodenum and ileum, whereas the total number of anaerobes increased only in duodenum. A significant reduction in bone strength and bone mineralization was observed in chickens fed with rye when compared with corn fed chickens. In conclusion, rye evoked mucosal damage in chickens that alter the intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition as well as bone mineralization. Studies to evaluate dietary inclusion of selected DFM candidates that produce exogenous enzymes in rye fed chickens are currently being evaluated