Next-generation non-starch polysaccharide-degrading, multi-carbohydrase complex rich in xylanase and arabinofuranosidase to enhance broiler feed digestibility.

This study was carried out to evaluate the effect of a multi-carbohydrase complex (MCC) rich in xylanase (Xyl) and arabinofuranosidase (Abf) on overall broiler feed digestibility in broilers. Energy utilization and digestibility of dry matter (DM), organic matter (OM), protein, starch, fat, and insoluble and soluble fibers were measured using the mass-balance method. The experiment was carried out on 120 broilers (3-week-old chickens). Broilers were distributed over 8 treatments to evaluate the effect of the dietary arabinoxylan content and nutrient density with and without MCC (Rovabio® Advance). The graded content of arabinoxylan (AX) was obtained using different raw materials (wheat, rye, barley, and dried distillers' wheat). Diet-energy density was modified with added fat. Measurements indicated that nutrient density and AX content had a significant effect on most digestibility parameters. Apparent metabolizable energy (AME) was significantly increased (265 kcal kg-1) by MCC. The addition of MCC also resulted in significant improvement in the digestibility of all evaluated nutrients, with average improvements of 3.0, 3.3, 3.2, 3.0, 6.2, 2.9, 5.8, and 3.8% units for DM, OM, protein, starch, fat, insoluble and soluble fibers, and energy utilization, respectively. The interaction between MCC and diet composition was significant for the digestibility of OM, fat, protein, and energy. Nutrient digestibility and diet AME were negatively correlated with AX content (P < 0.001). However, the addition of MCC resulted in a reduction of this negative effect (P < 0.001). The AME of diets with and without the addition of MCC were successfully predicted by the diet digestible nutrient (i.e., starch, protein, fat, insoluble and soluble fibers) content with and without MCC (R2 = 0.87; RSD = 78 kcal kg-1). This study confirms that the presence of AX in wheat-based diets and wheat-based diets with other cereals and cereal by-products reduces nutrient digestibility in broiler chickens. Furthermore, the dietary addition of MCC, which is rich in Xyn and Abf, reduced deleterious effect of fiber and improved overall nutrient digestibility in broiler diets.

Wheat-kernel-associated endoxylanases consist of a majority of microbial and a minority of wheat endogenous endoxylanases.

The endoxylanases associated with wheat kernels consist of wheat endogenous endoxylanases on one hand and kernel-associated microbial endoxylanases on the other hand. Assessment of their presence, based on analysis of their enzymic activity, can be expected to be hampered by the presence in wheat of high levels of endogenous endoxylanase inhibitors, which are able to inhibit the wheat-kernel-associated microbial endoxylanases. On the basis of preliminary experiments aimed at clarifying the distribution of the wheat-associated endoxylanases, a method to estimate total endoxylanase activities in wheat kernels was developed. Extensive washing of wheat kernels with universal buffer of pH 8.0 provided near-quantitative separation of the microbial endoxylanases located on the surface of wheat kernels from the endogenous endoxylanases and endoxylanase inhibitors located in such kernels. The microbial or endogenous nature of the endoxylanases was confirmed by making use of the inhibition specificity of endoxylanase inhibitors. Determination of the endoxylanase activity in the washing liquid, corresponding to the microbial endoxylanase population, and the washed kernels, corresponding to the endogenous endoxylanase population, allowed estimation of the total endoxylanase activities associated with the wheat kernel. Results showed that microbial endoxylanases can account for over 90% of the total wheat-associated endoxylanase activity and that the latter can be at least 5 times higher than the apparent endoxylanase activity.

Profile of enzyme production by Trichoderma reesei grown on corn fiber fractions.

Corn fiber is the fibrous by-product of wet-mill corn processing. It typically consists of about 20% starch, 14% cellulose, and 30% hemicellulose in the form of arabinoxylan. Crude corn fiber (CCF) was fractionated into de-starched corn fiber (DSCF), corn fiber with cellulose (CFC) enriched, and corn fiber arabinoxylan (CFAX), and these fractions were evaluated as substrates for enzyme production by Trichoderma reesei. T. reesei QM9414 and Rut C-30 grew on CCF, DSCF, CFC, or CFAX and secreted a number of hydrolytic enzymes. The enzymes displayed synergism with commercial cellulases for corn fiber hydrolysis.

Pre- and postweaning attributes in faunated and ciliate-free calves fed calf starter with or without fish meal.

In a 2 x 2 factorial design, 24 newborn, crossbred (Bos indicus x Bos taurus) calves were distributed in 4 equal groups involving dietary treatments of prestarter diets with (FM) or without fish meal (NFM) in a faunated (F) or ciliate-free (D) ruminal environment to study the ruminal fermentative development in pre-and postweaning periods. Defaunation was achieved by rearing calves in isolation and its effect was studied after first appearance of ciliate protozoa (observed after 8 wk of age) in the faunated animals. Calves were fed colostrum for 24 h and whole milk until weaning at 8 wk of age. Ruminal content samples were collected on d 4, 1 wk, weekly to 8 wk, and then biweekly at 9, 11, and 13 wk of age. The samples were analyzed for fermentation products [pH, total volatile fatty acids (VFA) and ammonia N] and enzyme [carboxymethyl (CM) cellulase, xylanase, beta-glucosidase, alpha-amylase, beta-galactosidase, proteases, and urease] activities. Weekly feed intake increased with age, but was similar in both groups. Ruminal pH declined steadily during 0 to 4 wk of age and then stabilized. The total VFA concentration increased with the age. The ammonia N (mg/dL) concentration increased from 14.9 on d 4 to 32.4 at 4 wk, decreased to 17.6 at 8 wk, and then steadied during the postweaning period. Samples collected on d 4 had no fibrolytic activity. Xylanase (U/dL) appeared first (1 wk) followed by beta-glucosidase (U/dL) and CM cellulase (U/dL), which increased steadily from a low of 4.69, 0.08, and 2.95 to 31.8 (6 wk), 5.92 (7 wk), and 19.8 (8 wk), respectively, and the concentrations showed nonsignificant alterations during postweaning periods. The concentration of alpha-amylase (U/dL) increased from 34.3 on d 4 to 87.2 at 8 wk, and then decreased to 56.6 (13 wk). beta-Galactosidase increased up to 6 wk then decreased to trace level (0.20 U/dL) at 13 wk of age. The concentrations of proteases and urease reached a steady state after 1 wk of age. The effect of diet type on ruminal fermentation products and enzyme parameters was nonsignificant. However, a steady and proportional alteration in both parameters in response to dry feed intake with the advancement of age was seen in all calves. Defaunation increased total VFA (97.3 vs. 75.8 mM/L) and alpha-amylase activity (80.3 vs. 61.4 U/dL) and decreased ammonia N (16.4 vs. 21.1 mg/dL), whereas the effect on other parameters was nonsignificant. Ruminal fermentative changes responded to dry feed intake, but did not differ in response to animal protein in prestarter diet.

Effect of cereal brans on Lentinula edodes growth and enzyme activities during cultivation on forestry waste.

AIMS: To develop strategies for increasing the growth of Lentinula edodes in eucalyptus residues. To this end, we have examined the effects of cereal brans additions on production of mycelial biomass and enzymes. METHODS AND RESULTS: Three isolates of the mushroom shiitake, L. edodes (Berk. Pegler), were evaluated for enzyme and ergosterol production on eucalyptus residue supplemented with 5, 10, 15 and 20% (w/w) of soya, wheat or rice brans. Nitrogen imput on eucalyptus residues accelerated mycelial growth by supplying the L. edodes with this limiting nutrient. High levels of enzymes activities were produced in eucalyptus residues supplemented by soya bran. Comparison of cellulose and xylanase production with manganese peroxidase (MnP) at 20% soya bran indicated that hydrolytic enzymes, but oxidative enzymes were reduced. CONCLUSIONS: Mycelial growth measurements revealed that eucalyptus residues supplemented with cereal brans supported fast growth of L. edodes, indicating that mycelium extension is related to the bioavailability of nitrogen. The type and concentration of nutrient supplement has a considerable effect both on substrate colonization and on the type of hydrolytic and oxidative enzymes produced. These characteristics may be useful for mushroom growing. SIGNIFICANCE AND IMPACT OF THE STUDY: Lentinula edodes is commercially important for edible mushroom production and supplements which enhance growth and enzymes production might also be beneficial for mushroom yields.

Quantification of supplemental enzymes in animal feedingstuffs by radial enzyme diffusion.

Methods are described which facilitate quantification of supplemental cellulase, protease and alpha-amylase when added to animal feedingstuffs at normal industrial inclusion levels. The methods entail extraction of the enzymes from the feedingstuffs by agitation in buffer followed by quantification of extract activity using radial diffusion techniques. A linear relationship between the diameter of the zone of hydrolyzed substrate and the log of the enzyme activity applied is observed over a broad activity range. Assay of a feedingstuff supplemented with 1 kg t(-1) cellulase, protease and alpha-amylase yielded net supplemental activity recoveries of 104+/-11.7%, 91.3+/-6.74% and 126+/-29.5%, respectively. A similar assay method did not prove sufficiently sensitive to facilitate detection of xylanase at typical in-feed inclusion levels. The levels of endogenous cellulase, protease and alpha-amylase activity detected in the unsupplemented feedingstuffs were equivalent to 6.4+/-0.47%, 6.6+/-0.82% and 29.0+/-14.1%, respectively, of a 1 kg t(-1) supplement. The methods are technically straightforward and will facilitate determination of enzyme stabilities during processes such as high-temperature pelleting of feedingstuffs, as well as allowing more rigorous quality control related to enzyme-supplemented animal feedingstuffs.