Lactational performance and nutrients digestibility response of dairy buffaloes fed diets supplemented with probiotic (Streptococcus spp.) and fibrolytic enzymes.

The current study was conducted to explore the productive performance and health status of lactating buffaloes fed diets supplemented with probiotic and/or fibrolytic enzymes. Forty multiparous lactating Egyptian buffaloes (body weight 451 ± 8.5 kg) were equally assigned to four experimental groups: (1) the first group fed control diet, (2) second experimental group fed control diet plus 4 g of probiotic/kg dry matter (DM) (probiotic), (3) third experimental group fed control diet plus 4 g of fibrolytic enzymes/kg DM (enzymes) and (4) fourth experimental group fed control diet plus 2 g of probiotic + 2 g fibrolytic enzymes/kg DM (Mix), The experiment was extended for 63 days. Nutrients digestibility was estimated, daily milk yield was recorded and milk samples were analyzed for total solids, fat protein, lactose and ash. Blood serum samples were analyzed for glucose, total protein, albumin, urea-N, aspartate transaminase, alanine transaminase and cholesterol concentrations. Results showed that adding probiotic and/or fibrolytic enzymes improved nutrients digestibility (p < 0.05). The probiotic, enzymes and mix groups did not affect (p > 0.05) concentrations of serum total protein, albumin (A), globulin (G), albumin/globulin (A/G) ratio and urea-N concentrations. An improvement in daily milk yield (p < 0.0001) and energy-corrected milk (p = 0.0146) were observed with the probiotic and mix groups compared with the control. In conclusion, this study suggests that supplementing lactating buffaloes' diets with probiotic alone or in combination with fibrolytic enzymes would improve their productive performance without adversely impacting their health.

Impacts of Solid-State Fermented Barley with Fibrolytic Exogenous Enzymes on Feed Utilization, and Antioxidant Status of Broiler Chickens.

The present and future high demand of common cereals as corn and wheat encourage the development of feed processing technology that allows for the dietary inclusion of other cereals of low nutritional value in poultry feeding. Barley grains contain anti-nutritional factors that limit their dietary inclusion in the poultry industry. The treatment of barley with solid-state fermentation and exogenous enzymes (FBEs) provides a good alternative to common cereals. In this study, barley grains were subjected to solid-state microbial fermentation using Lactobacillus plantarum, Bacillus subtilis and exogenous fibrolytic enzymes. This study aimed to assess the impact of FBEs on growth, feed utilization efficiency, immune modulation, antioxidant status and the expression of intestinal barrier and nutrient transporter-related genes. One-day-old broiler chicks (Ross 308, n = 400) comprised four representative groups with ten replicates (10 chicks/replicate) and were fed corn-soybean meal basal diets with inclusions of FBEs at 0, 5, 10 and 15% for 38 days. Solid-state fermentation of barley grains with fibrolytic enzymes increased protein content, lowered crude fiber and reduced sugars compared to non-fermented barley gains. In consequence, the group fed FBEs10% had the superior feed utilization efficiency and body weight gain (increased by 4.7%) with higher levels of nutrient metabolizability, pancreatic digestive enzyme activities and low digesta viscosity. Notably, the group fed FBEs10% showed an increased villi height and a decreased crypt depth with a remarkable hyperactivity of duodenal glands. In addition, higher inclusion levels of FBEs boosted serum immune-related parameters and intestinal and breast muscle antioxidants status. Intestinal nutrient transporters encoding genes (GLUT-1, CAAT-1, LAT1 and PepT-1) and intestinal barriers encoding genes (MUC-2, JAM-2, occludin, claudins-1 and ß-defensin 1) were upregulated with higher dietary FBEs levels. In conclusion, feeding on FBEs10% positively enhanced broiler chickens' performance, feed efficiency and antioxidant status, and boosted intestinal barrier nutrient transporters encoding genes.

Exogenous fibrolytic enzymes promoted energy and nitrogen utilization and decreased CH4 emission per unit dry matter intake of tan sheep grazed a typical steppe by enhancing nutrient digestibility on China loess plateau.

Exogenous fibrolytic enzyme (EFE) products in ruminant nutrition may be an important alternative to meet the increased demands for animal products in the future with reduced environmental impacts. This study aimed to evaluate the dose-response of EFE supplementation on the nutrient digestibility, nitrogen and energy utilization, and methane (CH4) emissions of Tan sheep grazed in summer and winter. A total of 20 Tan wether sheep with an initial body weight of 23.17 ±â€…0.24 kg were used in a randomized complete block design and categorized into two groups. Animals fed orally with 1 g of EFE (10,000 U/g) mixed with 30 mL of water using a drencher constituted the EFE group. For experimental accuracy, the control (CON) group was orally administered with 30 mL of normal saline daily before grazing. The following results were obtained: EFE in the diet increased dry matter intake (DMI) (P < 0.05), average daily gain (ADG) (P < 0.05), and digestibility (P < 0.05) compared with CON in summer and winter. DMI increased but ADG and digestibility decreased in winter compared with those in summer. Sheep fed with the EFE diet increased the concentrations of rumen ammonia nitrogen (P < 0.05) and total volatile fatty acids (P > 0.05), but reduced pH (P > 0.05), compared with CON in summer and winter. EFE increased nitrogen (N) intake, digestible N, retained N, and retained N/digestible N (P < 0.05) but reduced fecal N/N intake, urinary N/N intake, and excretion N/N intake in summer and winter (P < 0.05), compared with CON. Retained N/N intake was reduced and excretion N/N intake increased in winter relative to those in summer. In winter, gross energy (GE), manure E/GE, CH4 emissions, CH4/DMI, and CH4/GE increased but digestion energy and metabolic energy decreased compared with those in summer. Sheep fed with the EFE diet had a greater GE intake than those fed with the CON diet (P < 0.05) but had lesser CH4/DMI and CH4E/GE (P < 0.05) than those fed with the CON diet in both summer and winter. In conclusion, EFE supplementation increased DMI, apparent digestibility, and N deposition rate. These effects were beneficial for animal production. The CH4 emission per unit DMI of grazing Tan sheep was lesser and conducive for augmenting the environmental benefits.

Globally, the supply­demand relationship between grassland and livestock is mainly mediated by the optimization of pasture management. The interaction between grassland and livestock is one of the fundamental drivers of grassland occurrence and development. Natural grassland yields and quality are affected by precipitation, heat, and grazing, and their dynamics vary seasonally with distinct peaks and troughs. The use of exogenous fibrolytic enzymes during troughs can improve the growth performance, digestion, and metabolism of grazing sheep. The exogenous fibrolytic enzyme supplement used in this research may aid in improving the health and overall productivity of grazing sheep.

Utilization of exogenous fibrolytic enzymes in fiber fermentation, degradation, and digestions and characteristics of whole legume faba bean and its plant silage.

This article aims to review recent progress and update on utilization of exogenous fibrolytic enzymes in fiber fermentation, degradation, and digestions and nutritive and anti-nutritional characteristics of whole legume faba bean and its silage. The study focused on strategies to improve the utilization and bioavailability of fiber through pre-treating exogenous fibrolytic enzymes. The review includes features of nutrition and anti-nutritional factors and environment impact, forage fiber fermentation, degradation and digestion, legume bean in various diets, use of exogenous enzyme and factor affecting enzyme action in fiber digestion as well as exogenous enzyme response. This review also provides very recent research on effects of fibrolytic enzyme on rumen degradation characteristics of dry matter and fiber of whole plant faba bean silage and effect of exogenous fibrolytic enzyme derived from trichoderma reesei on lactational performance, feeding behavior, rumen fermentation and nutrient digestibility in dairy cows fed whole plant faba bean silage-based diet. This study provides an insight on nutritive and anti-nutritive characteristics of whole legume bean and its plant silage and utilization of exogenous fibrolytic enzymes in fiber fermentation, degradation, and digestions.

Improvement of Ruminal Neutral Detergent Fiber Degradability by Obtaining and Using Exogenous Fibrolytic Enzymes from White-Rot Fungi.

The present review examines the factors and variables that should be considered to obtain, design, and evaluate EFEs that might enhance ruminal NDF degradability. Different combinations of words were introduced in Google Scholar, then scientific articles were examined and included if the reported factors and variables addressed the objective of this review. One-hundred-and-sixteen articles were included. The fungal strains and culture media used to grow white-rot fungi induced the production of specific isoforms of cellulases and xylanases; therefore, EFE products for ruminant feed applications should be obtained in cultures that include the high-fibrous forages used in the diets of those animals. Additionally, the temperature, pH, osmolarity conditions, and EFE synergisms and interactions with ruminal microbiota and endogenous fibrolytic enzymes should be considered. More consistent results have been observed in studies that correlate the cellulase-to-xylanase ratio with ruminant productive behavior. EFE protection (immobilization) allows researchers to obtain enzymatic products that may act under ruminal pH and temperature conditions. It is possible to generate multi-enzyme cocktails that act at different times, re-associate enzymes, and simulate natural protective structures such as cellulosomes. Some EFEs could consistently improve ruminal NDF degradability if we consider fungal cultures and ruminal environmental conditions variables, and include biotechnological tools that might be useful to design novel enzymatic products.

The effectiveness of fibrolytic enzymes and active yeast on improving reticulorumen pH in dairy cows.

Exogenous fibrolytic enzymes (EFE) and yeast are feed supplements that improve forage digestion in rumen, but their influences on physical reticulorumen parameters are not well studied. This study was designed to evaluate the effect of the EFE:endo-ß-xylanase (37x104 U/cow/day), endocellulase (45x104 U/cow/day), endo-ß-glucanase (12x104U/cow/day), and active yeast - Saccharomyces cerevisiae CNCM-1077 (10x109CFU/cow/day) supplements on reticulorumen pH (RpH) and temperature (RT) in dairy cows. Nine Lithuanian Red cows were allocated into three groups (3 cows/group): control group (C) - farm diet without supplementation, enzyme group (E) - farm diet supplemented with EFE, enzyme and active yeast group (EY) - farm diet supplemented with EFE and active yeast. The feeding trial lasted for 60 d. All cows were equipped with reticuloruminal telemetric pH and temperature sensor device. Data provided by the device were used to calculate the mean RpH (RpH/24h), the mean minimal RpH (minRpH/24h) and mean of the time that RpH was below the threshold value of 6.0 (RpH⟨6.0/24h, min.). The highest RpH/24h (6.37±0.22) was observed in group EY and it was by 1.62% (p⟨0.05) and 1.27% (p⟨0.001) higher as compared with groups E and C, respectively. Also minRpH/24h (6.24±0.24) was highest in group EY and values were by 0.63% (p⟨0.001) and 0.65% (p⟨0.001) higher as compared with groups C and E, respectively. The shortest duration of RpH⟨6.0/24h, was recorded in group EY, and it was by 57.76% (p⟨0.05) and 47.87% shorter as compared with groups C and E, respectively. In conclusion, feed supplementation with EFE and Saccharomyces cerevisiae CNCM-1077 had beneficial effect on RpH.

Effect of exogenous fibrolytic enzymes and ammonia fiber expansion on the fermentation of wheat straw in an artificial rumen system (RUSITEC)1.

This study investigated the effect of treatment of wheat straw using ammonia fiber expansion (AFEX) and exogenous fibrolytic enzymes (Viscozyme) on fiber digestibility, rumen fermentation, microbial protein synthesis, and microbial populations in an artificial rumen system [Rumen Simulation Technique (RUSITEC)]. Four treatments were assigned to 16 vessels (4 per treatment) in 2 RUSITEC apparatuses in a randomized block design. Treatments were arranged as a 2 × 2 factorial using untreated or AFEX-treated wheat straw with or without exogenous fibrolytic enzymes [0 or 500 µg of protein/g straw dry matter (DM)]. Fibrolytic enzymes were applied to straw, prior to sealing in nylon bags. The concentrate mixture was provided in a separate bag within each fermentation vessel. The RUSITECs were adapted for 8 d and disappearance of DM, neutral detergent fiber (NDF), acid detergent fiber (ADF), and crude protein (CP) was measured after 48 h of incubation. Ammonia fiber expansion increased (P < 0.01) the disappearance of wheat straw DM (69.6 vs. 38.3%), NDF (65.6 vs. 36.8%), ADF (61.4 vs. 36.0%), and CP (68.3 vs. 24.0%). Total dietary DM, organic matter (OM), and NDF disappearance was also increased (P ≤ 0.05) by enzymes. Total microbial protein production was greater (P < 0.01) for AFEX-treated (72.9 mg/d) than untreated straw (63.1 mg/d). Total gas and methane (CH4) production (P < 0.01) were also greater for AFEX-treated wheat straw than untreated straw, with a tendency for total gas to increase (P = 0.06) with enzymes. Ammonia fiber expansion increased (P < 0.01) total volatile fatty acid (VFA) production and the molar proportion of propionate, while it decreased (P < 0.01) acetate and the acetate-to-propionate ratio. The AFEX-treated straw had lower relative quantities of fungi, methanogens, and Fibrobacter succinogenes (P < 0.01) and fewer protozoa (P < 0.01) compared to untreated straw. The pH of fermenters fed AFEX-treated straw was lower (P < 0.01) than those fed untreated straw. Both AFEX (P < 0.01) and enzymes (P = 0.02) decreased xylanase activity. There was an enzyme × straw interaction (P = 0.02) for endoglucanase activity. Enzymes increased endoglucanase activity of AFEX-treated wheat straw, but had no effect on untreated straw. The addition of enzymes lowered the relative abundance of Ruminococcus flavefaciens, but increased F. succinogenes. These results indicate that AFEX increased the ruminal disappearance of wheat straw and improved fermentation and microbial protein synthesis in the RUSITEC.

New recombinant fibrolytic enzymes for improved in vitro ruminal fiber degradability of barley straw1.

This study used a high-throughput in vitro microassay, in vitro batch culture, and the Rumen Simulation Technique (RUSITEC) to screen recombinant fibrolytic enzymes for their ability to increase the ruminal fiber degradability of barley straw. Eleven different recombinant enzymes in combination with a crude mixture of rumen enzymes (50% recombinant enzyme:50% crude mixture of rumen enzymes) were compared with the crude mixture of rumen enzymes alone. In the microassay, all treatments were applied at 15 mg of protein load per gram barley straw glucan. Based on the microassay results, 1 recombinant endoglucanase [EGL7A, from the glycoside hydrolase (GH) family 7], 2 recombinant xylanases (XYL10A and XYL10C, from GH10), and a recombinant enzyme mixture were selected and compared with a crude mixture of fibrolytic enzymes from Aspergillus aculeatus for their ability to hydrolyze barley straw. For batch culture, enzymes were applied to barley straw at 2 dosages (100 and 500 µg of protein/g of substrate DM). All enzymes increased (P < 0.05) DM disappearance and total VFA production, but the mixture of recombinant enzymes was not superior to the use of a single recombinant enzyme. Based on positive results (P < 0.05) for total DM disappearance and VFA production in batch culture, 3 enzymes (EGL7A, XYL10A, and XYL10C) were selected and applied to barley straw at 500 µg of protein per gram for further assessment in RUSITECs fed a concentrate:barley straw diet (300:700 g/kg DM). In RUSITECs, the recombinant enzyme XYL10A increased (P < 0.05) barley straw DM, NDF, and ADF disappearance, whereas EGL7A and XYL10C had no effect. The enzymes selected based on the high-throughput in vitro microassay consistently increased barley straw degradation in ruminal batch culture, but not in the semicontinuous culture RUSITEC system.

In vitro evaluation of total mixed ration supplemented with exogenous fibrolytic enzymes for crossbred cows.

AIM: The study was conducted to evaluate the levels of exogenous fibrolytic enzymes (EFE) on in vitro digestibilities of dry matter (DM) and organic matter (OM), total gas production (TGP), metabolizable energy (ME) content, and microbial biomass production (MBP). MATERIALS AND METHODS: The total mixed ration (TMR) was prepared using 30% each of sorghum hay and groundnut straw and 40% compound concentrate mixture to meet nutritional requirement of cow (500 kg) producing 12 kg fat corrected milk. The EFE was incorporated at 0, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, and 400 mg/kg TMR. The TMR substrates with different levels of EFE were in vitro incubated to ascertain their effect on digestibility, gas production, and nutritive values. RESULTS: The significantly (p<0.05) higher and optimum in vitro digestibilities of DM (63.03%) and OM (63.62%) as well as TGP (72.35 ml/500 mg TMR) were observed at supplementation of 240 mg EFE/kg TMR, while ME (7.16 MJ/kg DM) and MBP (97.63 mg/500 mg TMR) were also better. CONCLUSION: The incorporation of EFE at 240 mg/kg TMR resulted significantly (p<0.05) higher and optimum in vitro digestibilities of DM and OM. The TGP, ME, and MBP were also better. The levels of EFE 240 mg/kg TMR were found suitable for further in vivo study in crossbred cows.

A meta-analysis on the effect of dietary application of exogenous fibrolytic enzymes on the performance of dairy cows.

The aim of this study was to use meta-analytical methods to estimate effects of adding exogenous fibrolytic enzymes (EFE) to dairy cow diets on their performance and to determine which factors affect the response. Fifteen studies with 17 experiments and 36 observations met the study selection criteria for inclusion in the meta-analysis. The effects were compared by using random-effect models to examine the raw mean difference (RMD) and standardized mean difference between EFE and control treatments after both were weighted with the inverse of the study variances. Heterogeneity sources evaluated by meta-regression included experimental duration, EFE type and application rate, form (liquid or solid), and method (application to the forage, concentrate, or total mixed ration). Only the cellulase-xylanase (C-X) enzymes had a substantial number of observations (n = 13 studies). Application of EFE, overall, did not affect dry matter intake, feed efficiency but tended to increase total-tract dry matter digestibility and neutral detergent fiber digestibility (NDFD) by relatively small amounts (1.36 and 2.30%, respectively, or <0.31 standard deviation units). Application of EFE increased yields of milk (0.83 kg/d), 3.5% fat-corrected milk (0.55 kg/d), milk protein (0.03 kg/d), and milk lactose (0.05 kg/d) by moderate to small amounts (<0.30 standard deviation units). Low heterogeneity (I 2 statistic <25%) was present for yields and concentrations of milk fat and protein and lactose yield. Moderate heterogeneity (I 2 = 25 to 50%) was detected for dry matter intake, milk yield, 3.5% fat-corrected milk, and feed efficiency (kg of milk/kg of dry matter intake), whereas high heterogeneity (I 2 > 50%) was detected for total-tract dry matter digestibility and NDFD. Milk production responses were higher for the C-X enzymes (RMD = 1.04 kg/d; 95% confidence interval: 0.33 to 1.74), but were still only moderate, about 0.35 standardized mean difference. A 24% numerical increase in the RMD resulting from examining only C-X enzymes instead of all enzymes (RMD = 1.04 vs. 0.83 kg/d) suggests that had more studies met the inclusion criteria, the C-X enzymes would have statistically increased the milk response relative to that for all enzymes. Increasing the EFE application rate had no effect on performance measures. Application of EFE to the total mixed ration improved only milk protein concentration, and application to the forage or concentrate had no effect. Applying EFE tended to increase dry matter digestibility and NDFD and increased milk yield by relatively small amounts, reflecting the variable response among EFE types.

Trends in recombinant protein use in animal production.

Recombinant technologies have made possible the production of a broad catalogue of proteins of interest, including those used for animal production. The most widely studied proteins for the animal sector are those with an important role in reproduction, feed efficiency, and health. Nowadays, mammalian cells and fungi are the preferred choice for recombinant production of hormones for reproductive purposes and fibrolytic enzymes to enhance animal performance, respectively. However, the development of low-cost products is a priority, particularly in livestock. The study of cell factories such as yeast and bacteria has notably increased in the last decades to make the new developed reproductive hormones and fibrolytic enzymes a real alternative to the marketed ones. Important efforts have also been invested to developing new recombinant strategies for prevention and therapy, including passive immunization and modulation of the immune system. This offers the possibility to reduce the use of antibiotics by controlling physiological processes and improve the efficacy of preventing infections. Thus, nowadays different recombinant fibrolytic enzymes, hormones, and therapeutic molecules with optimized properties have been successfully produced through cost-effective processes using microbial cell factories. However, despite the important achievements for reducing protein production expenses, alternative strategies to further reduce these costs are still required. In this context, it is necessary to make a giant leap towards the use of novel strategies, such as nanotechnology, that combined with recombinant technology would make recombinant molecules affordable for animal industry.

Metagenomic insights into the rumen microbial fibrolytic enzymes in Indian crossbred cattle fed finger millet straw.

The rumen is a unique natural habitat, exhibiting an unparalleled genetic resource of fibrolytic enzymes of microbial origin that degrade plant polysaccharides. The objectives of this study were to identify the principal plant cell wall-degrading enzymes and the taxonomic profile of rumen microbial communities that are associated with it. The cattle rumen microflora and the carbohydrate-active enzymes were functionally classified through a whole metagenomic sequencing approach. Analysis of the assembled sequences by the Carbohydrate-active enzyme analysis Toolkit identified the candidate genes encoding fibrolytic enzymes belonging to different classes of glycoside hydrolases(11,010 contigs), glycosyltransferases (6366 contigs), carbohydrate esterases (4945 contigs), carbohydrate-binding modules (1975 contigs), polysaccharide lyases (480 contigs), and auxiliary activities (115 contigs). Phylogenetic analysis of CAZyme encoding contigs revealed that a significant proportion of CAZymes were contributed by bacteria belonging to genera Prevotella, Bacteroides, Fibrobacter, Clostridium, and Ruminococcus. The results indicated that the cattle rumen microbiome and the CAZymes are highly complex, structurally similar but compositionally distinct from other ruminants. The unique characteristics of rumen microbiota and the enzymes produced by resident microbes provide opportunities to improve the feed conversion efficiency in ruminants and serve as a reservoir of industrially important enzymes for cellulosic biofuel production.

Fribrinolytic activity and gas production by Pleurotus ostreatus-IE8 and Fomes fomentarius - EUM1 in bagasse cane / Actividad fibrolítica y producción de gas por Pleurotus ostreatus-IE8 y Fomes fomentarius-EUM1 en bagazo de caña

Objective. To characterize the fibrolytic enzymatic activity of Pleurotus ostreatus-IE8 and Fomes fomentarius-EUM1 in sugarcane bagasse (BCA); to evaluation of the kinetics of in vitro production of BCA treated by solid fermentation (FS), crude enzyme extract (ECE) of P. ostreatus-IE8 and Fibrozyme®. Materials and methods. In fungi measured radial growth rate ( Vcr ) and biomass production in two culture media (with or without nitrogen source); activity of xylanases, cellulases and FS on BCA at 0, 7 and 15 d. The chemical analysis and kinetic analysis of in vitro gas production in 4 treatments (ECE adding enzymes obtained from the direct addition FS or FS ), witness (Fibrozyme®) and a control without addition and analyzed by a was completely randomized design. Results. Xylanases (7 d ) showed 6.32 and 5.50 UI g-1 initial substrate dry weight (SSi) for fungi P. ostreatus-IE8 and F. fomentarius-EUM1 , respectively ; P. ostreatus-IE8 scored higher activity of laccases (10.65 g-1 UI SSi) and F. fomentarius-EUM1 (1.90 UI g-1 SSi) cellulases. The ECE of P. ostreatus-IE8 and commercial enzyme did not differences (p>0.05). In the chemical composition or the gas production kinetics. The 4 treatments evaluated decreased values of the variables measured in the kinetics of gas production compared to the control (p≤0.05). Conclusions. The ECE of P. ostreatus-IE8 was similar to commercial enzyme degradation in vitro, so it is feasible to use pre-digest high fiber products.

Objetivos. Caracterizar la actividad enzimática fibrolítica de Pleurotus ostreatus-IE8 y Fomes fomentarius-EUM1 en bagazo de caña de azúcar (BCA) y evaluar la cinética de producción de gas in vitro del BCA por fermentación sólida (FS) o con extractos crudos enzimáticos (ECE) de P. ostreatus-IE8 o Fibrozyme®. Materiales y métodos. En los hongos de estudio se evaluó la velocidad de crecimiento radial (Vcr) y producción de biomasa en dos medios de cultivo (con o sin fuente de nitrógeno); actividad de xilanasas, celulasas y lacasas de la FS sobre BCA a 0, 7 y 15 d. El análisis químico y cinética de producción de gas in vitro en 4 tratamientos (proceso de FS o adición de enzimas obtenidas de ECE de la FS), un testigo (Fibrozyme®) y un control sin adición de enzimas, todo ello se analizó en un diseño completamente al azar. Resultados. Las xilanasas (7 d) mostraron 6.32 y 5.50 UI g-1 sustrato seco inicial (SSi) en P. ostreatus-IE8 y F. fomentarius-EUM1, respectivamente. P. ostreatus-IE8 mostró mayor actividad lacasa (10.65 UI g-1 SSi) y F. fomentarius-EUM1 (1.90 UI g-1 SSi) de celulasas. El ECE de P. ostreatus-IE8 y Fibrozyme® no presentaron diferencias (p>0.05) en la composición química ni en la cinética de producción de gas. Los 4 tratamientos evaluados disminuyeron los valores de las variables medidas en la cinética de producción de gas in vitro respecto al testigo (p≤0.05). Conclusiones. El ECE de P. ostreatus-IE8 fue similar a Fibrozyme® en la degradación in vitro, indicando su viabilidad y uso para pre-digerir subproductos altos en fibra.

Effect of exogenous fibrolytic enzymes on performance and blood profile in early and mid-lactation Holstein cows.

The supplementation of exogenous fibrolytic enzymes (EFE) to dairy cows diets could be a strategy to improve fiber degradation in the rumen which is especially important for the early lactating cows characterized by a high milk energy output and an insufficient energy intake. The objective of this study was to examine the effects of a fibrolytic enzyme product (Roxazyme G2 Liquid, 3.8 and 3.9 mL/kg total mixed ration [TMR] DM) supplemented to a TMR on production performance and blood parameters of dairy cows during early (trial 1) and mid-lactation (trail 2). In addition, rumination activity was measured in trial 2. The nutrient digestibility of the experimental TMR was obtained by using wethers. In the digestibility trial, EFE was supplemented at a rate of 4.4 mL/kg Roxazyme G2 Liquid TMR-DM. The TMR contained 60% forage and 40% concentrate (DM basis). Twenty eight 50 ± 16 days in milk (DIM) and twenty six 136 ± 26 DIM Holstein cows were used in two 8-wk completely randomized trails, stratified by parity and milk yield level. One milliliter of the enzyme product contained primarily cellulase and xylanase activities (8,000 units endo-1,4-ß glucanase, 18,000 units endo-1,3(4)-ß glucanase and 26,000 units 1,4-ß xylanase). No differences in digestibility of DM, OM, CP, NDF and ADF were observed (P > 0.05) between the control and the EFE supplemented TMR. Addition of EFE to the TMR fed to early (trial 1) and mid-lactation cows (trial 2) did not affect daily dry matter intake (DMI), milk yield, 4% fat-corrected milk, energy-corrected milk (ECM), concentration of milk fat, protein, fat-protein-quotients, somatic cell score, energy balance, and gross feed efficiency of early and mid-lactation cows (P > 0.05). Mid-lactation cows (trial 2) fed with TMR enzyme showed a tendency of a slightly higher ECM yield (P = 0.09). The tested blood parameters were not affected by treatment in trials 1 and 2 (P > 0.05). Exogenous fibrolytic enzymes supplementation did not alter daily time spent ruminating in trial 2 (P = 0.44). In conclusion, under the conditions of this study, no positive effects of enzyme supplementation on dairy performance and health status of dairy cows during early and mid-lactation were observed.

Treatment of tropical forages with exogenous fibrolytic enzymes: effects on chemical composition and in vitro rumen fermentation.

The effects of three treatments of fibrolytic enzymes (cellulase from Trichoderma longibrachiatum (CEL), xylanase from rumen micro-organisms (XYL) and a 1:1 mixture of CEL and XYL (MIX) on the in vitro fermentation of two samples of Pennisetum clandestinum (P1 and P2), two samples of Dichanthium aristatum (D1 and D2) and one sample of each Acacia decurrens and Acacia mangium (A1 and A2) were investigated. The first experiment compared the effects of two methods of applying the enzymes to forages, either at the time of incubation or 24 h before, on the in vitro gas production. In general, the 24 h pre-treatment resulted in higher values of gas production rate, and this application method was chosen for a second study investigating the effects of enzymes on chemical composition and in vitro fermentation of forages. The pre-treatment with CEL for 24 h reduced (p < 0.05) the content of neutral detergent fibre (NDF) of P1, P2, D1 and D2, and that of MIX reduced the NDF content of P1 and D1, but XYL had no effect on any forage. The CEL treatment increased (p < 0.05) total volatile fatty acid (VFA) production for all forages (ranging from 8.6% to 22.7%), but in general, no effects of MIX and XYL were observed. For both P. clandestinum samples, CEL treatment reduced (p < 0.05) the molar proportion of acetate and increased (p < 0.05) that of butyrate, but only subtle changes in VFA profile were observed for the rest of forages. Under the conditions of the present experiment, the treatment of tropical forages with CEL stimulated their in vitro ruminal fermentation, but XYL did not produce any positive effect. These results showed clearly that effectiveness of enzymes varied with the incubated forage and further study is warranted to investigate specific, optimal enzyme-substrate combinations.

Development of appropriate fibrolytic enzyme combination for maize stover and its effect on rumen fermentation in sheep.

In vitro studies were undertaken to develop an appropriate fibrolytic enzymes cocktail comprising of cellulase, xylanase and ß-D-glucanase for maize stover with an aim to increase its nutrient utilization in sheep. Cellulase and xylanase added individually to ground maize stover at an increasing dose rates (0, 100, 200, 400, 800, 1,600, 3,200, 6,400, 12,800, 25,600, 32,000, 38,400, and 44,800 IU/g DM), increased (p<0.01) the in vitro dry matter digestibility and in vitro sugar release. The doses selected for studying the combination effect of enzymes were 6,400 to 32,000 IU/g of cellulase and 12,800 to 44,800 IU/g of xylanase. At cellulase concentration of 6,400 IU/g, IVDMD % was higher (p<0.01) at higher xylanase doses (25,600 to 44,800 IU/g). While at cellulase doses (12,800 to 32,000 IU/g), IVDMD % was higher at lower xylanase doses (12,800 and 25,600 IU/g) compared to higher xylanase doses (32,000 to 44,800 IU/g). At cellulase concentration of the 6,400 to 32,000 IU/g, the amount of sugar released increased (p<0.01) with increasing levels of xylanase concentrations except for the concentration of 44,800 IU/g. No effect of ß-D-glucanase (100 to 300 IU/g) was observed at lower cellulase-xylanase dose (cellulase-xylanase 12,800 to 12,800 IU/g). Based on the IVDMD, the enzyme combination cellulase-xylanase 12,800 to 12,800 IU/g was selected to study its effect on feed intake and rumen fermentation pattern, conducted on 12 rams (6 to 8 months; 20.34±2.369 kg body weight) fed 50% maize stover based TMR. The total volatile fatty acids (p<0.01) and ammonia-N concentration was higher in enzyme supplemented group, while no effect was observed on dry matter intake, ruminal pH and total nitrogen concentration.