Metabolizable energy in corn is greater than in hybrid rye when fed to gestating sows, but exogenous enzymes did not increase energy digestibility

The metabolizable energy (ME) in corn was greater (P < 0.05) than in hybrid rye when fed to sows in mid-gestation, although the difference between the two grains was less than the difference previously observed in growing pigs. An exogenous enzyme mixture did not increase apparent total tract digestibility of dry matter or gross energy in corn or hybrid rye fed to gestating sows. Likewise, digestible energy and ME in corn and hybrid rye were not increased by the enzymes.

β-Mannanase Supplementation as an Eco-Friendly Feed Strategy to Reduce the Environmental Impacts of Pig and Poultry Feeding Programs

Little is still known about the environmental impacts of exogenous enzyme supplementation in pig and poultry feeding programs. Thus, this study aimed to assess the potential environmental impacts of producing feeds for pigs and broilers by simulating the effects of β-mannanase Hemicell™ HT supplementation through energy savings during diet formulation. Life-cycle assessment standards were applied to simulate a cradle-to-feed mill gate scope. The functional units used were the production of 1 kg of the enzyme and 1 kg of feed at a feed mill gate located in Concórdia, Santa Catarina, Brazil. Climate change, eutrophication, and acidification were the chosen environmental impact categories. Energy savings through β-mannanase supplementation were assessed by different metabolizable energy (ME) matrices (45 or 90 kcal of ME/kg of feed) during diet formulation in different grain production scenarios (Southern and/or Central-West origin). A total of 28 feeds were formulated based on the nutritional requirements and feeding programs described in the Brazilian Tables for Poultry and Swine. The least-cost formulation method was used based on real price averages practiced in a local industry over 12 months. The production of 1 kg of β-mannanase was associated with the emission of 1,800 g of CO2-eq, 4.53 g of PO4-eq, and 7.89 g of SO2-eq. For pig feeds, β-mannanase supplementation mitigated both climate change and eutrophication impacts up to 8.5 and 1.4% (45 kcal of ME/kg of feed) or up to 16.2 and 2.7% (90 kcal of ME/kg of feed) compared to control diets formulated without the enzyme. For broiler feeds, these impacts were mitigated up to 5.6 and 1.1% (45 kcal of ME/kg of feed), respectively. On the other hand, the effect of using β-mannanase on the acidification impact was not consistent among feeds/species. Overall, β-mannanase supplementation reduced the amount of soybean oil in feed formulas, which is associated with high environmental impacts. Consequently, the potential impacts of climate change and eutrophication associated with producing feeds for pigs and broilers were substantially mitigated. These results suggest that β-mannanase supplementation is an eco-friendly feed strategy to reduce the environmental impacts of pig and poultry feeding programs.

Characterisation of Extracellular Enzyme-Producing Microorganisms From the Gut of Nile Tilapia, Oreochromis niloticus (Linnaeus, 1758)

The current priority in the aquaculture industry is to replace fish meal with cheaper but efficient protein sources like plant-based feedstuffs to support global expansion and sustainability. Hence, research has focused on finding novel ways to increase the digestibility of plant-based diets. Exogenous enzymes have emerged as an excellent alternative to improve nutrient digestibility of the diet in animals, including fish. The present study aims to explore exogenous enzyme-producing bacteria in the gastrointestinal (GI) tract of Nile tilapia (Oreochromis niloticus (Linnaeus, 1758)). Two Gram-negative enzyme-producing (amylase, xylanase, and lipase) bacterial strains viz. Aeromonas veronii ONKP1 (GenBank accession no. MN602971) and Stenotrophomonas maltophilia ONKP2 (GenBank accession no. MN602972) were identified by biochemical tests and 16s RNA analysis. They are capable of utilising citrate, maltose, glucose, and gelatine. Besides, A. veronii ONKP1 can utilise mannitol, sucrose, and lactose, whereas S. maltophilia ONKP2 is catalase and urease positive. Aeromonas veronii ONKP1 was superior in terms of enzyme production to S. maltophilia ONKP2. Stenotrophomonas maltophilia ONKP2 is a rarely reported strain, specifically from healthy fish. Amylase, xylanase, and lipase could be used as feed additives for fermenting plant products and producing prebiotics like xylooligosaccharides. Further, this study might help understand the role of gut-associated bacteria viz. A. veronii and S. maltophilia, in fish nutrition.

Effect of Exogenous Enzymes on Egg Fertility Traits, Egg Production and Oviduct Micro-structure of Layer Birds

The effect of two classes of exogenous enzymes on the egg fertility traits, egg production efficiency and oviduct micro-structure of laying hens was evaluated. Forty-five laying hens (30-weeks old, 1.32-1.40 kg) were randomly allotted into three dietary treatments with fifteen birds per treatment (five birds each in three replicates per treatment) in a completely randomized design. Diet one served as the control diet. Diet two (FS diet) contained a nutrient-targeted exogenous enzyme preparation (Fullzyme®) at 200 g/ton, while diet three (DS diet) contained a mycotoxin-targeted exogenous enzyme preparation (Detoxizyme®) at 500 g/ton. Data on fertility traits (egg volume, length and breadth, weight, yolk diameter, albumen height, air cell, shell thickness, Haugh’s unit, and egg-lay), and egg production (feed intake, egg lay, egg lay efficiency, and feed cost per egg laid) were collected for 42 days. At the end of the experiment, three randomly selected birds per group were euthanized, and their oviducts were harvested and processed for histopathology. Hens fed enzyme supplemented diets had significantly (p < 0.05) better egg fertility traits. Feed intake in the control group did not vary (p > 0.05)from feed intake in the DS or FS group. However, feed cost per egg laid and egg-lay efficiency was higher in the control diet group than in the FS group or DS diet group. No pathologies were observed in the oviduct of birds fed enzymes supplemented or control diet. Exogenous enzymes in diet improved egg fertility traits and production without altering the oviduct micro-structure of Layer hens.

Innovative strategies to enhance the sensory quality of dry fermented sausages containing lactic ingredients by the addition of exogenous enzymes

This study investigated the impact of the addition of exogenous enzymes (Accelerzyme CPG, Debitrase DBP20) or cellular preparations (FlavoGard), traditionally used in the cheese industry, to accelerate flavour development of dry fermented sausages with 6% of lactic derivatives content. Sausages were fermented to pH 5.0, dried for 32 days and vacuum packed stored under refrigeration for 60 days. Sausages were analysed for physicochemical parameters, technological microbiota and proteolysis after fermentation, drying/ripening and storage. Similar compositional results were obtained in all products (38-39% humidity in the final product; 38.2% fat and 40.7% protein as dry matter throughout the study). Debitrase application positively affected proteolysis by changing the free amino acid profile and increasing non-protein nitrogen and total free amino acids by 2.2 and 11.8-fold, respectively. Accelerzyme increased ripened cheese flavour and overall sensory quality from 5.1 to 5.8; Debitrase increased ripened cheese odour and flavour, bitterness, umami, adhesiveness, pastiness, and overall sensory quality from 5.0 to 5.9, and decreased acid and hardness. This study highlights the effects of adding some exogenous enzyme/bacterial preparations traditionally used in the cheese industry to enhance the flavour of dry fermented sausages with high content of lactic ingredients and increase its sensory quality.

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.

Autophagy modulation enhances enzyme replacement therapy response in Fabry disease

Fabry disease is a lysosomal disease due to α-galactosidase A (a-GalA) deficiency. Since 2001, Enzyme replacement therapy (ERT) has been used as specific treatment of Fabry disease, with variable effects depending on patient gender and affected organs. In Fabry cells, the endososo-mal/lysosomal system is highly altered. Consequently, the exogenous enzyme may be mistargeted and trapped into intracellular vesicles instead of reaching the lysosomes. In this study, we aimed to investigate the mechanisms underlying the processing of the exogenous enzyme. We used Fabry cells (cultured fibroblasts and podocyte cell line) to study the enzyme internalization and its effects on the catabolism of the main a-Gal A substrate, globotriaosylceramide (Gb3), upon autophagy inhibition. The exogenous enzyme reaches the early endosome in a similar timeframe in Fabry and control cells, while its targeting to lysosomes is delayed in Fabry cells. Gb3 concentration is lowered upon therapeutic enzyme addition or wortmannin treatment with a synergetic effect. These findings illustrate the positive impact of autophagy inhibition on enzyme trafficking and processing, allowing the increase of functional enzyme rate within the lysosome. Given the high cost of ERT, a better understanding of the cellular fate of the exogenous enzyme may lead to improve its targeting to the lysosome.

Coccidia Vaccine Challenge and Exogenous Enzyme Supplementation in Broiler Chicken 1. Effect on Digesta Viscosity, Diet Energy Utilization, and Apparent Metabolizable Energy Value of Wheat

This study examined the effect of exogenous mixed-enzyme supplementation (xylanase, β-glucanase, and pectinase) to a corn-SBM (CS) and a wheat-CS-based (WCS) diet in birds challenged with coccidia vaccine (Coccivac B-52™). The WCS-based diet was produced by replacing 30% of the energy-yielding portions of the CS-based diet with wheat. On day 14, 448 (n = 7) Cobb by-product breeder male broilers were assigned to a 2 (diet types) × 2 (with or without enzyme supplementation) × 2 (0 or 20 × coccidia vaccine challenge; CVC) factorial arrangement of treatments in a completely randomized design for the determination of the apparent metabolizable energy (AME) value of wheat. Treatment effects on jejunum digesta viscosity and AME corrected for nitrogen (AMEn) of the diets were evaluated within each diet type as a 2 × 2 factorial arrangement of treatments, 7- and 14-day post-challenge. Seven-day post-challenge (day 21), dry matter (DM) and energy utilization, AME, and AMEn of the CS- and WCS-based diets decreased (p < 0.05) with CVC. Both AME and AMEn of wheat decreased (p < 0.05) by about a 20% in CVC-birds 7-day post-challenge. Enzyme and CVC resulted in a decrease (p < 0.05) in jejunal digesta viscosity in birds fed the CS-based diets, while there was an interaction (p < 0.05) between CVC and enzyme, with enzyme lowering (p < 0.05) the viscosity of digesta 7-day post-challenge. Results from this study showed that CVC resulted in a 20% decrease in AMEn 7-day post-challenge, while the interaction between exogenous enzyme supplementation and CVC resulted in an improvement in nitrogen utilization (~6%) in CVC birds fed the corn-SBM-based diet 7-days post challenge.

Gut morphology, meat yield and sensory properties of laying hens fed water hyacinth {Eichhornia crassipes (Mart.) Solms-Laubach} meal diets supplemented with Maxigrain® enzyme

In an experiment to determine the effect of feeding varying levels of water Hyacinth Meal (WHM) supplemented with and without exogenous enzymes (Maxigrain®) on the gut morphology, meat yield and sensor properties of laying hens, a total of 216 laying hens aged 24 weeks and 4 weeks in lay were used. Six experimental diets were formulated based on the Completely Randomized Design model using a 2r3 factorial arrangement made up of no levels of enzyme (No enzyme, With enzyme) and three dietary inclusion levels of WHM (0.10 and 20%). At the end of the feeding trial, two birds per replicate were randomly selected, kept of feed for 12 hours, slaughtered, defeathered, eviscerated and dressed. The meat of the breast and thigh of each replicate sample were then selected for sensory evaluation. All parameters were expressed as percentage of live weight. Results showed that there were no significant (p>0.05) differences in live weight(LW), slaughter weight (SW), defeathered weight(DIN), eviscerated weight (EW) and dressed weight (DW) between birds fed diets supplemented with exogenous enzymes and those without exogenous enzyme supplementation and between birds fed the varying dietary inclusion levels of WHM (0,10 and 20%). Also, there were no significant (p>0.05) differences in the weight of the head, neck, breast, back, thigh, drumsticks, wings and shanks between the birds fed diets supplemented with exogenous enzymes and those without exogenous enzyme supplementation; and between the birds fed the varying dietary inclusion levels of WHM (0. 10 and 20% ). Of all the internal organs assessed, only the gizzard weight was significantly (P<0.05) higher for the non-enzyme-supplemented diets than for the enzyme-supplemented diets. There were no significant (p>0.05) differences in appearance, flavour juiciness, tenderness and overall acceptability between the meat of birds fed diets supplemented with exogenous enzymes and those without exogenous enzyme supplementation, but the meat of birds fed 10 and 20% dietary inclusion levels of WHM had significantly (p<0.05) higher scores for tenderness and general acceptability than the meat of birds fed the 0% dietary inclusion levels of WHM. Hence, WHM can be included up to 20% in layer diets for optimal meat yield, gut morphology and sensory properties.