Multi-Enzyme Supplementation Modifies the Gut Microbiome and Metabolome in Breeding Hens

Laying and reproductive performance, egg quality, and disease resistance of hens decrease during the late laying period. Exogenous enzymes promote nutrient digestibility and utilization and improve the intestinal environment. However, the specific regulation of the gut microbiome and metabolome by exogenous enzymes remains unelucidated. This study was conducted to evaluate effects of dietary multi-enzyme supplementation on egg and reproductive performance, egg quality, ileum microbiome, and metabolome of breeders. Here, 224 Hy-Line Brown breeding hens (55 weeks old) were randomly allocated to two groups: dietary controls fed basal diet (DC), and test hens fed 0.2 g/kg corn enzyme diet (CE). Serum levels of total protein, globulin, immunoglobulin Y, and antibodies against the Newcastle disease virus and avian influenza H9 strain were significantly increased (p < 0.05). Egg albumen height, Haugh unit, and fertilization and hatching rates were also significantly increased (p < 0.05) in the CE-fed group. 16S rRNA sequence analysis showed that CE strongly affected both α- and β-diversity of the ileal microbiota. LEfSe analysis revealed that the potentially beneficial genera Lactobacillus, Enterococcus, Faecalicoccus, and Streptococcus were enriched as biomarkers in the CE-fed group. Microbial functional analysis revealed that the functional genes associated with harmful-substance biodegradation was significantly increased in the CE-fed group. Additionally, Spearman correlation analysis indicated that changes in microbial genera were correlated with differential metabolites. In summary, dietary multi-enzyme addition can improve egg quality, humoral immunity, and reproductive performance and regulate the intestinal microbiome and metabolome in breeders. Therefore, multi-enzymes could be used as feed additive to extend breeder service life.

P-OGC03 Pancreatic enzyme supplementation improves quality of life in patients following surgery for upper GI cancer

Background Unpleasant abdominal symptoms are common following surgery for upper gastrointestinal (UGI) cancer and may occur secondary to pancreatic exocrine insufficiency (EPI). This study investigated symptoms of EPI in patients following surgery and assessed the effect of pancreatic enzyme supplementation (PERT) on these symptoms and the effect of supplementation on quality of life. Methods Patients were assessed for symptoms of EPI using a novel questionnaire. Patients who reported two or more symptoms suggestive of EPI were prescribed PERT. Abdominal symptoms were reassessed following treatment. Quality of life (QoL) was studied using the SF-36 questionnaire before and after treatment. Faecal elastase was also measured in a patient subgroup. Results Fifty-six out of 57 patients (98%) reported at least two symptoms of EPI. Following PERT every patient reported fewer abdominal symptoms; median 5 symptoms before treatment reduced to two symptoms following treatment (p < 0.0001; Wilcoxon rank). Reduced faecal elastase concentration was associated with more frequent abdominal symptoms; median 5 symptoms versus 3 symptoms (p = 0.043; Mann Whitney U test). PERT increased quality of life scores for every patient in each of the 5 principle health domains. Conclusions Symptoms of EPI are common among patients following UGI cancer surgery. PERT reduces unpleasant abdominal symptoms and this leads to significant improvements in quality of life across global health domains. PERT should be offered to all post-operative UGI cancer patients with symptoms suggestive of EPI.

Greenhouse gases and performance of growing pigs fed wheat-based diets containing wheat millrun and a multi-carbohydrase enzyme

The objective of this project was to determine the impact of feeding growing pigs with high wheat millrun diets supplemented with a multi-carbohydrase enzyme (amylase, cellulase, glucanase, xylanase, and invertase activities) on nutrient digestibility, growth performance, and greenhouse gas (GHG) output (carbon dioxide, CO2; nitrous oxide, N2O; methane, CH4). Three experiments were conducted utilizing six treatments arranged as a 3 × 2 factorial (0%, 15%, or 30% wheat millrun; with or without enzyme) for the digestibility experiment or as a 2 × 2 factorial (0% or 30% wheat millrun; with or without enzyme) for the performance and GHG experiments. The digestibility, performance, and GHG experiments utilized 48 individually housed pigs, 180 pigs housed 5 per pen, or 96 pigs housed 6 per chamber, respectively. Increasing wheat millrun up to 30% in the diet of growing pigs resulted in decreased energy, nitrogen (N) and phosphorus (P) apparent total tract digestibility and net energy content (P < 0.01). Overall, average daily gain (ADG) and gain to feed ratio were reduced in pigs fed wheat millrun (P < 0.05). Enzyme supplementation had minimal effects on the digestibility or performance parameters measured. Feeding diets with 30% millrun did not affect GHG output (CH4: 4.7 and 4.9; N2O: 0.45 and 0.42; CO2: 1,610 and 1,711 mg/s without or with millrun inclusion, respectively; P > 0.78). Enzyme supplementation had no effect on GHG emissions (CH4: 4.5 and 5.1; N2O: 0.46 and 0.42; CO2: 1,808 and 1,513 mg/s without or with enzymes, respectively; P > 0.51). Overall, the carbohydrase enzyme had minimal effects on parameters measured, regardless of wheat millrun inclusion (P > 0.10). Although energy, N and P digestibility, and ADG were reduced, the inclusion of up to 30% wheat millrun in the diet has no effect on GHG emissions from growing pigs (P > 0.10).

Broiler feed formulated with phosphorus, calcium, and energy deficiencies supplemented with exogenous enzymes

Three experiments were carried out to verify the effects of the enzyme phytase, alone or combined with an enzyme complex, in diets deficient in available phosphorus (AP), calcium (Ca), and metabolizable energy (ME) on broiler performance, ME, and dietary amino acid digestibility. A total of 1,538 male Cobb 500 broilers were allocated to the three experiments, each of which consisted of five treatments: positive control (PC; basal ration); negative control 1 (NC1; PC minus 0.15% of AP, 0.16% of Ca, and 68 kcal kg-1 ME); negative control 2 (NC2; PC minus 0.15% of AP, 0.16% of Ca, and 101 kcal kg-1 of ME); NC1 plus phytase; and NC2 plus phytase plus enzymatic complex. Body weight gain (WG) and feed intake were measured from 1-21 days and from 1-42 days, and the corrected feed conversion rate (FCR) for mortality was calculated. In the second and third experiments, the apparent ME corrected for nitrogen balance (AMEn) and standardized digestibility of amino acids, respectively, were determined, for the diets supplemented with phytase and the enzymatic complex. In the first experiment, enzyme supplementation increased (p < 0.05) WG at 21 days and 42 days relative to the negative controls. Phytase inclusion improved (p < 0.05) FCR at the initial phase compared to the NC1 diet. In the second experiment, enzyme supplementation did not affect (p > 0.05) AMEn. In the third experiment, both enzyme treatments improved (p < 0.05) the digestibility of amino acids in the supplemented diets compared to the deficient diets. Supplementation with phytase and carbohydrases preserves the performance of broilers fed diets deficient in AP, Ca, and ME and improves amino acid digestibility.

Impact of Dietary or Drinking Water Ruminococcus sp. Supplementation and/or Heat Stress on Growth, Histopathology, and Bursal Gene Expression of Broilers

This research was conducted to evaluate the impact of dietary or drinking water Ruminococcus sp. supplementation and/or heat stress (HS) on the growth, serum biochemistry, tissue antioxidant, phagocytic assay, histopathology, and bursa gene expression of broilers. Day-old broiler chicks were allotted into six groups according to HS and/or Ruminococcus with or without enzyme supplementation. The first group was the control one, with a formulated diet and normal environmental temperature but without any supplement. The second group fed on Ruminococcus-supplemented diet (1 kg/kg diet). The third group fed on a formulated diet without supplement, and Ruminococcus and digestive enzymes were given in drinking water (0.1 ml/L). The fourth one was the heat stress group, with a normal formulated diet. The fifth and the sixth groups served as second and third groups, respectively, but with heat stress. The results of this experiment indicated that thermal temperature negatively affected the parameters of growth performance, serum biochemical, tissue antioxidants, and phagocytic assay. Moreover, heat stress led to pathological lesions in the internal organs and affected the expression of some genes related to heat stress, including proapoptotic genes such as caspase8 and bax, inflammatory genes such as NF-κβ1, and heat shock protein such as HSP 70 in the bursal tissue. These bad effects and abnormalities were mitigated by Ruminococcus alone or with enzyme supplementation, which improved all the above-mentioned parameters.

196 Effect of Exogenous Enzyme Supplementation and Fermentation of Oilseed By-products on in vitro Digestibility and Production of Short Chain Fatty Acid

An experiment was conducted to investigate the effect of exogenous enzyme supplementation and solid-state fermentation (SSF) with a mixed bacterial culture on in vitro dry matter digestibility (IVDMD) and short chain fatty acid (SCFA) production of soybean meal (SBM) or rapeseed meal (RSM). A 2 × 2 factorial design was used and included the factors of 1) exogenous enzyme cocktail (supplemented and non-supplemented), 2) microbial fermentation (fermented and non-fermented) applied to SBM or RSM in vitro. The exogenous enzyme cocktail consisted of non-starch polysaccharide (NSP) degrading enzymes (NSP-EZ) with phytase (10,000 FTU/kg), and the SSF were carried out using Bacillus subtilis. The fermented feed ingredients were collected after 48 h incubation at 37oC. Samples were hydrolyzed in two steps using pepsin and pancreatin to calculate IVDMD. Subsequently, the hydrolyzed residues were filtered, dried, and pooled for incubation in a buffered mineral solution with fresh swine feces. Gas production kinetics during fermentation was measured for 72 h and analyzed by fitting data to an exponential model. The fermentation residues were filtered, and the supernatant was analyzed for concentration of SCFA. The IVDMD from simulated gastric and small intestinal hydrolysis was greater (P &lt; 0.01) for SSF in both SBM and RSM. During fermentation, the hydrolysis residue from SBM treated with SSF required less time to reach half asymptote, had greater maximal gas production, and greater fractional degradation (P &lt; 0.01, respectively) compared with non-fermented SBM. The IVDMD from simulated total tract digestion was greater (P &lt; 0.01) for SSF in RSM compared with SBM, while SBM had greater IVDMD for both SSF and NSP-EZ (P &lt; 0.01). Production of butyric acid was greater for SSF (P &lt; 0.01) compared with non-SSF in both SBM and RSM. These results suggest that SSF can improve IVDMD and produce greater amounts of butyric acid compared with NSP-EZ supplementation in SBM and RSM.

Impact of Xylanase and Glucanase on Oligosaccharide Formation, Carbohydrate Fermentation Patterns, and Nutrient Utilization in the Gastrointestinal Tract of Broilers

This study aimed at determining how the degradation of cereal non-starch polysaccharides (NSP) by dietary enzymes during feed digestion can influence nutrient digestibility and NSP fermentability in broilers. Ninety-six one-day-old male broilers were assigned to 4 different treatments: control and enzyme-supplemented wheat-based (WC, WE) or maize-based (MC, ME) treatments. Enzyme supplementation with endo-xylanase and endo-glucanase occurred from day 20 onwards. On day 28, digesta samples were collected. Nutrient digestibility, NSP recovery, oligosaccharide profile, and short-chain fatty acids (SCFA) content were determined. Enzyme supplementation in WE resulted in a higher starch (3%; p = 0.004) and protein (5%; p = 0.002) digestion in the ileum compared to WC. Xylanase activity in WE led to in situ formations of arabinoxylan-oligosaccharides consisting of 5 to 26 pentose units in the ileum. This coincided with decreased arabinose (p = 0.059) and xylose (p = 0.036) amounts in the ceca and higher acetate (p = 0.014) and butyrate (p = 0.044) formation in WE compared to WC. Conversely, complete total tract recovery of arabinoxylan in MC and ME suggested poor maize NSP fermentability. Overall, enzyme action improved nutrient digestibility and arabinoxylan fermentability in the wheat-based diet. The lower response of the maize-based diet to enzyme treatment may be related to the recalcitrance of maize arabinoxylan as well as to the high nutritive value of maize.