[Expression of ß-xylosidase An-xyl from Aspergillus niger and characterization of its xylose tolerance].

The hydrolysis of xylo-oligosaccharides catalyzed by ß-xylosidase plays an important role in the degradation of lignocellulose. However, the enzyme is easily inhibited by its catalytic product xylose, which severely limits its application. Based on molecular docking, this paper studied the xylose affinity of Aspergillus niger ß-xylosidase An-xyl, which was significantly differentially expressed in the fermentation medium of tea stalks, through cloning, expression and characterization. The synergistic degradation effect of this enzyme and cellulase on lignocellulose in tea stems was investigated. Molecular docking showed that the affinity of An-xyl to xylose was lower than that of Aspergillus oryzae ß-xylosidase with poor xylose tolerance. The Ki value of xylose inhibition constant of recombinant-expressed An-xyl was 433.2 mmol/L, higher than that of most ß-xylosidases of the GH3 family. The Km and Vmax towards pNPX were 3.6 mmol/L and 10 000 µmol/(min·mL), respectively. The optimum temperature of An-xyl was 65 ℃, the optimum pH was 4.0, 61% of the An-xyl activity could be retained upon treatment at 65 ℃ for 300 min, and 80% of the An-xyl activity could be retained upon treatment at pH 2.0-8.0 for 24 h. The hydrolysis of tea stem by An-xyl and cellulase produced 19.3% and 38.6% higher reducing sugar content at 2 h and 4 h, respectively, than that of using cellulase alone. This study showed that the An-xyl mined from differential expression exhibited high xylose tolerance and higher catalytic activity and stability, and could hydrolyze tea stem lignocellulose synergistically, which enriched the resource of ß-xylosidase with high xylose tolerance, thus may facilitate the advanced experimental research and its application.

Soil enzyme responses to land use change in the tropical rainforest of the Colombian Amazon region.

Soil enzymes mediate key processes and functions of the soils, such as organic matter decomposition and nutrient cycling in both natural and agricultural ecosystems. Here, we studied the activity of five extracellular soil enzymes involved in the C, N, and P-mineralizing process in both litter and surface soil layer of rainforest in the northwest region of the Colombian Amazon and the response of those soil enzymes to land use change. The experimental study design included six study sites for comparing long-term pasture systems to native forest and regeneration practices after pasture, within the main landscapes of the region, mountain and hill landscapes separately. Results showed considerable enzymatic activity in the litter layer of the forest, highlighting the vital role of this compartment in the nutrient cycling of low fertility soils from tropical regions. With the land use transition to pastures, changes in soil enzymatic activities were driven by the management of pastures, with SOC and N losses and reduced absolute activity of soil enzymes in long-term pastures under continuous grazing (25 years). However, the enzyme activities expressed per unit of SOC did not show changes in C and N-acquiring enzymes, suggesting a higher mineralization potential in pastures. Enzymatic stoichiometry analysis indicated a microbial P limitation that could lead to a high catabolic activity with a potential increase in the use of SOC by microbial communities in the search for P, thus affecting soil C sequestration, soil quality and the provision of soil-related ecosystem services.

Exogenous enzymes and probiotics alter digestion kinetics, volatile fatty acid content and microbial interactions in the gut of Nile tilapia.

Sustainable aquafeed production requires fishmeal replacement, leading to an increasing use of plant-derived ingredients. As a consequence, higher levels of antinutritional substances, such as non-starch polysaccharides and phytate, are present in aquafeeds, with negative effects on fish performance, nutrient digestibility and overall gut health. To alleviate these negative effects, providing exogenous digestive enzymes and/or probiotics can be an effective solution. In this study, we tested the effect of dietary supplementation of enzymes (phytase and xylanase) and probiotics (three strains of Bacillus amyloliquefaciens) on nutrient digestion kinetics and volatile fatty acid content along the gut, and the distal gut microbiome diversity in Nile tilapia. Chyme volatile fatty content was increased with probiotic supplementation in the proximal gut, while lactate content, measured for the first time in vivo in fish, decreased with enzymes along the gut. Enzyme supplementation enhanced crude protein, Ca and P digestibility in proximal and middle gut. Enzymes and probiotics supplementation enhanced microbial interactions as shown by network analysis, while increased the abundance of lactic acid bacteria and Bacillus species. Such results suggest that supplementation with exogenous enzymes and probiotics increases nutrient availability, while at the same time benefits gut health and contributes to a more stable microbiome environment.

Aroma enhancement of instant green tea infusion using ß-glucosidase and ß-xylosidase.

ß-Glucosidase and ß-xylosidase were investigated for their ability to improve the aroma of instant green tea. The aroma and corresponding contributors were analyzed by sensory evaluation, gas chromatography-mass spectrometry, and odor activity value. Their specific contributions to aroma attributes were further examined by aroma reconstruction and omission experiments. The ß-glucosidase treatment significantly enhanced floral and grassy notes, on account of the increases of geraniol, nonanal, and cis-3-hexen-1-ol, and weakened the caramel note, attributable to the increases of nonanal, cis-3-hexen-1-ol, geraniol, methyl salicylate, and decanal. The co-treatment with ß-glucosidase and ß-xylosidase further enhanced the grassy note, with further increase in nonanal and cis-3-hexen-1-ol, and further weakened the caramel note, with additional increase in nonanal, cis-3-hexen-1-ol, methyl salicylate, and decanal. The synergistic action of ß-glucosidase and ß-xylosidase provides new clues to the production of instant green tea infusions with high aroma quality.

Development of novel economical methodologies for zymographic analysis of purified xylano-pectinolytic enzymes using agrowaste-based substrates.

In this study, zymographic analysis for xylanase and pectinase enzymes has been carried out using agrowaste residues, wheat bran and citrus peel as well as their extracts. Isozymic forms of xylanase as well as pectinase enzyme displayed comparable zymographic bands onto agar petriplates containing either commercial substrates (xylan and pectin), agrowaste-based substrates (wheat bran and citrus peel), or polysaccharides extracted from these agrowastes (crude xylan and pectin extracted from wheat bran and citrus peel, respectively), indicating the fact that agro residues and their extracts can be utilized as a substitute of cost-intensive commercial substrates, xylan and pectin for zymographic analysis. This is the first report revealing the zymographic analysis of xylano-pectinolytic enzymes using agro-based solid residues particles or polysaccharides extracted from agro-based residues.

Exogenous fibrolytic enzymes and recombinant bacterial expansins synergistically improve hydrolysis and in vitro digestibility of bermudagrass haylage.

Four experiments were conducted to examine the effects of a recombinant bacterial expansin-like protein (BsEXLX1) from Bacillus subtilis and a commercial exogenous fibrolytic enzyme (EFE) preparation for ruminants on hydrolysis of pure substrates (cellulose and xylan) and in vitro digestibility of bermudagrass haylage (BMH). Recombinant Escherichia coli BL21 strain was used to express BsEXLX1; the protein was purified using an affinity column. In experiment 1, carboxymethylcellulose, Whatman #1 filter paper (General Electric, Boston, MA) and oat-spelt xylan substrates were subjected to 4 treatments (1) sodium citrate buffer (control), (2) BsEXLX1 (162 µg/g of substrate), (3) EFE (2.3 mg/g of substrate), and (4) EFE + BsELX1 in 3 independent runs. Samples were incubated at optimal conditions for both additives (pH 5 and 50°C) or at ruminal (pH 6 and 39°C) or ambient (pH 6 and 25°C) conditions for 24 h and sugar release was measured. In experiment 2, digestibility in vitro of BMH was examined after treatment with the following: (1) control (buffer only), (2) BsEXLX1 (162 µg/g of dry matter), (3) EFE (2.2 mg/g of dry matter), and (4) EFE + BsEXLX1 in 3 independent runs at 39°C for 24 h. Experiment 3 examined effects of EFE and BsEXLX1 on simulated preingestive hydrolysis and profile of released sugars from BMH after samples were suspended in deionized water with sodium azide at 25°C for 24 h in 2 independent runs. In experiment 4, the sequence of the BsEXLX1 purified protein was compared with 447 ruminal bacterial genomes to identify similar proteins from the rumen. In experiment 1, compared with EFE alone, EFE and BsEXLX1 synergistically increased sugar release from carboxymethylcellulose and Whatman #1 filter paper under all simulated conditions; however, hydrolysis of xylan was not improved. In experiment 2, compared with EFE alone, treatment with EFE and BsEXLX1 increased neutral detergent fiber and acid detergent fiber digestibility of bermudagrass haylage (by 5.5 and 15%, respectively) and total volatile fatty acid concentrations, and decreased acetate-propionate ratio. In experiment 3, compared with EFE alone. The EFE and BsEXLX1 synergistically reduced concentrations of neutral detergent fiber and acid detergent fiber and increased release of sugars by 9.3%, particularly cellobiose (72.5%). In experiment 4, a similar sequence to that of BsEXLX1 was identified in Bacillus licheniformis, and similar hypothetical protein sequences were identified in Ruminococcus flavefaciens strains along with different protein structures in E. xylanophilum and Lachnospiraceae. This study showed that an expansin-like protein synergistically increased the hydrolysis of pure cellulose substrates and the hydrolysis and digestibility in vitro of BMH.

Effects of a combination of xylanase, amylase and protease, and probiotics on major nutrients including amino acids and non-starch polysaccharides utilization in broilers fed different level of fibers.

This study evaluated the effects of a combination of xylanase, amylase, and protease (XAP), with probiotics (3 Bacillus spp.) supplementation on apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in Cobb 500 broilers from 0 to 21 d. A completely randomized 2 × 4 factorial design (2 levels of fiber; 4 types of supplements) with 8 replicate cages (6 birds/cage) was used. Each low and high-fiber diet contained 500 FTU/kg Buttiauxella sp. phytase and was supplemented with: (a) none (control), (b) XAP (2,000 U xylanase + 200 U amylase + 4,000 U protease/kg diet), (c) probiotics (75,000 CFU/g of Bacillus spp.), or (d) XAP + probiotics. High fiber decreased (P < 0.05) nitrogen-corrected apparent metabolizable energy (AMEn), AID of all amino acids (AA), AID and ATTD of dry matter (DM), crude protein (CP), starch, and gross energy (GE). High fiber increased (P < 0.01) the flow of total non-starch polysaccharides (NSP) in both ileum and total tract. The XAP + probiotics increased (P < 0.01) AMEn as well as AID and ATTD of DM, CP, GE, starch, while alone, XAP yielded similar improvement except for DM compared with control. The supplemental XAP alone improved (P < 0.01) the digestibility of most of the AAs compared with control. Moreover, XAP + probiotics increased (P < 0.05) AID of all AA except arginine and serine compared with control. A fiber × supplements interaction (P < 0.05) was found for AID of histidine and threonine, and their digestibility in high-fiber diet was improved to a level comparable to low-fiber diet by XAP + probiotics. The flow of NSP in XAP group was 5 to 6% lower than in control while NSP flow in XAP + probiotic group was further 4% lower than that of XAP group (P < 0.01). The results infer that the combination of XAP and probiotics can effectively optimize the nutrient digestibility in broilers fed both low and high-fiber diets.

A novel and cost-effective methodology for enhanced production of industrially valuable alkaline xylano-pectinolytic enzymes cocktail in short solid-state fermentation cycle.

The aim of this study was to enhance the production of xylano-pectinolytic enzymes concurrently and also to reduce the fermentation period. In this study, the effect of agro-residues extract-based inoculum on yield and fermentation time of xylano-pectinolytic enzymes was studied. Microbial inoculum and fermentation media were supplemented with xylan and pectin polysaccharides derived from agro-based residues. Enzymes production parameters were optimized through two-stage statistical design approach. Under optimized conditions (temperature 37°C, pH 7.2, K2 HPO4 0.22%, MgSO4 0.1%, gram flour 5.6%, substrate: moisture ratio 1:2, inoculum size 20%, agro-based crude xylan in production media 0.45%, and agro-based crude xylan-pectin in inoculum 0.13%), nearly 28,255 ± 565 and 9,202 ± 193 IU of xylanase and pectinase, respectively, were obtained per gram of substrate in a time interval of 6 days only. The yield of both xylano-pectinolytic enzymes was enhanced along with a reduction of nearly 24 h in fermentation time in comparison with control, using polysaccharides extracted from agro-residues. The activity of different types of pectinase enzymes such as exo-polymethylgalacturonase (exo-PMG), endo-PMG, exo-polygalacturonase (exo-PG), endo-PG, pectin lyase, pectate lyase, and pectin esterase was obtained as 1,601, 12.13, 5637, 24.86, 118.62, 124.32, and 12.56 IU/g, respectively, and was nearly twofold higher than obtained for all seven types in control samples. This is the first report mentioning the methodology for enhanced production of xylano-pectinolytic enzymes in short solid-state fermentation cycle using agro-residues extract-based inoculum and production media.

Immobilization of Aspergillus quadrilineatus RSNK-1 multi-enzymatic system for fruit juice treatment and mannooligosaccharide generation.

Aspergillus quadrilineatus RSNK-1 produced a multi-enzymatic system containing (U/gds) ß-mannanase (1021), endo-xylanase (1 9 1), α-galactosidase (3.42), ß-xylosidase (0.07) and ß-glucosidase (0.28) on low-cost copra meal (CM) in SSF. The enzyme preparation was covalently immobilized on aluminum oxide pellets (3 mm) under statistically optimized conditions leading to 73.17% immobilization yield. The immobilized enzyme (Man-AOP) displayed enhanced thermal and pH stability. Man-AOP was characterized by FTIR, SEM and PXRD revealing a covalent interaction. The bio-conjugate was successfully recycled for mannooligosaccharide (MOS) generation from locust bean gum (LBG) up to 10 cycles, yielding an average of 0.95 mg MOS/cycle. Man-AOP was also effective in clarification of apple, kiwi, orange and peach juices and enhanced their reducing sugar content. The bio-conjugate was useful in generation of MOS from mannan and enrichment of fruit juices.

Optimization of Cellulase and Xylanase Productions by Streptomyces thermocoprophilus Strain TC13W Using Oil Palm Empty Fruit Bunch and Tuna Condensate as Substrates.

The modified medium composed of the alkaline-pretreated oil palm empty fruit bunch (APEFB) and tuna condensate powder was used for cellulase and xylanase productions by Streptomyces thermocoprophilus strain TC13W. The APEFB contained 74.46% (w/w) cellulose, 15.72% (w/w) hemicellulose, and 6.40% (w/w) lignin. The tuna condensate powder contained 55.49% (w/w) protein and 11.05% (w/w) salt. In the modified medium with only 6.75 g/l tuna condensate powder, 10 g/l APEFB, and 0.5 g/l Tween 80, S. thermocoprophilus strain TC13W produced cellulase 4.9 U/ml and xylanase 9.0 U/ml. The enzyme productions in the modified medium were lower than cellulase (6.0 U/ml) and xylanase (12.0 U/ml) productions in the complex medium (CaCl2 0.1, MgSO4·7H2O 0.1, KH2PO4 0.5, K2HPO4 1.0, NaCl 0.2, yeast extract 5.0, NH4NO3 1.0, Tween 80 0.5). When tuna condensate powder in the modified medium was reduced to 5.0 g/l and Tween 80 was increased to 1.5 g/l, S. thermocoprophilus strain TC13W produced cellulase and xylanase activities of 9.1 and 12.1 U/ml, respectively. This study shows that the cost of enzyme production could be reduced by using pretreated EFB and tuna condensate as a carbon and a nitrogen source, respectively.

Development of strategy for simultaneous enhanced production of alkaline xylanase-pectinase enzymes by a bacterial isolate in short submerged fermentation cycle.

The aim of this study is to enhance the production of industrially valuable xylanase and pectinase enzymes in short duration, using agrowaste extracted substrates. Conventional cum statistical multifactor analysis approaches were used in order to evaluate the effect of crude extracted substrates, supplemented for the production of xylanase-pectinase enzymes. Incorporation of crude extracted xylan (1.2 mg/ml of inoculum) and pectin (4.8 mg/ml of inoculum) substrates in inoculum resulted in maximal xylanase (320 ± 15) and pectinase titre (90 ± 8) after 48 h, using 2% wheat bran and 2% citrus peel in production medium with 48 h of fermentation time, with one variable factor at a time approach. The best condition obtained after performing statistical multifactor interaction analysis includes 5.50 mg/ml of pectin in inoculum,1.50 mg/ml of xylan in inoculum, wheat bran 3%, temperature 37.5 °C, time 48 h, 7 mg/ml of pectin in production medium, peptone 1.05%, inoculum size 2% and inoculum age of 20 h, with alkaline xylanase activity of 415.22 ± 18.50 IU/ml and alkaline pectinase activity of 109.10 ± 8.80 IU/ml. Activity of different pectinolytic enzymes per ml was also calculated, with 18.98 IU of exo-polymethylgalacturonase, 0.14 IU of endo-polymethylgalacturonase, 80 IU of exo-polygalacturonase, 0.28 IU of endo-polygalacturonase, 1.42 IU of polymethylgalacturonate lyase, 1.47 IU of polygalacturonate lyase, 0.15 IU of pectin esterase. This is the first report mentioning the utilization of crude extracted xylan and extracted pectin in inoculum to get the increment in the activity of both alkaline xylanase-pectinase enzymes simultaneously under short submerged fermentation cycle.

Comparison of Mono- and Dikaryotic Medicinal Mushrooms Lignocellulolytic Enzyme Activity.

Mono- and dikaryotic medicinal mushroom strains isolated from four wood-rotting basidiomycete fruiting bodies were comparatively evaluated for laccase, manganese peroxidase, cellulase, and xylanase activities in submerged cultivation in glucose or mandarin peel-containing media. Mandarin peels appeared to be the preferred growth substrate for laccase production by both mono- and dikaryotic Trametes multicolor 511 and T. versicolor 5 while glucose favored laccase activity secretion by Pleurotus ostreatus 2175. Lignocellulose-deconstructing enzyme profiles were highly variable between the studied monokaryotic and dikaryotic strains. A distinctive superiority of enzyme activity of the dikaryotic Trametes versicolor 5 and P. ostreatus 2175 over the same species monokaryotic isolates was revealed. By contrast, laccase, cellulase, and xylanase activities of the monokaryotic strain of T. multicolor 511 were rather higher than those in the dikaryotic culture. At the same time, hydrolases activity of Schizophyllum commune 632 was practically independent of the origin of the fungal culture. The results suggest that the monokaryotic isolates derived from the basidiomycetes fruiting bodies inherit parental properties but the capacity of individual monokaryotic cultures to produce lignocellulose-deconstructing enzymes can vary considerably.

Linkages between straw decomposition rate and the change in microbial fractions and extracellular enzyme activities in soils under different long-term fertilization treatments.

In order to study the linkages between straw decomposition rate and the change in soil biological properties after straw addition to different fertilized soils, we collected soils from three long-term fertilization treatments (no-fertilizer, CK; nitrogen, phosphorus, and potassium fertilizers, NPK; NPK plus straw (S), NPKS), and incubated maize straw with these soils at 25°C for 75 days. The average straw carbon dioxide (CO2) emission rate in the CK+straw (S), NPK+S, and NPKS+S treatments was 0.58±0.51, 0.66±0.53, and 0.74±0.58 µg C g-1soil h-1, respectively. The average increase in the contents of fungi, bacteria, and Actinomycetes under straw addition treatments than the control soils (CK, NPK, and NPKS, respectively) changed in the order of CK+S≤NPK+S

Effects of heat treatment on enzyme activity and expression of key genes controlling cell wall remodeling in strawberry fruit.

Modification of cell wall polymers composition and structure is one of the main factors contributing to textural changes during strawberry (Fragaria x ananassa, Duch.) fruit ripening and storage. The present study aimed to provide new data to understand the molecular basis underlying the postharvest preservation of strawberry cell wall structure by heat treatment. Ripe fruit (cv. Aroma) were heat-treated in air oven (3 h at 45 °C) and then stored 8 days at 4 °C + 2 days at 20 °C, while maintaining a set of non-treated fruit as controls. The effect of heat stress on the expression pattern of key genes controlling strawberry cell wall metabolism, as well as some enzymatic activities was investigated. The expression of genes proved to be relevant for pectin disassembly and fruit softening process (FaPG1, FaPLB, FaPLC, FaAra1, FaßGal4) were down-regulated by heat treatment, while the expression of genes being involved in the reinforcement of cell wall as pectin-methylesterase (FaPME1) and xyloglucan endo-transglycosilase (FaXTH1) was up-regulated. Total cell wall amount as well as cellulose, hemicellulose, neutral sugars and ionically and covalently bounded pectins were higher in heat-stressed fruit compared to controls, which might be related to higher firmness values. Interestingly, heat stress was able to arrest the in vitro cell wall swelling process during postharvest fruit ripening, suggesting a preservation of cell wall structure, which was in agreement with a lower growth rate of Botrytis cinerea on plates containing cell walls from heat-stressed fruit when compared to controls.

Effect of Mediterranean saltbush (Atriplex halimus) ensilaging with two developed enzyme cocktails on feed intake, nutrient digestibility and ruminal fermentation in sheep.

The aim of this study was to assess the effects of feeding Atriplex halimus (AH) silage treated with two developed enzyme cocktails to sheep on feed intake, nutrient digestibility and ruminal fermentation. The AH silage was treated without or with 2 L of ZAD1(®) or ZAD2(®) /1000 kg with 5% molasses and ensiled for 30 days. Barley grain (300 g/head/day) was fed as an energy supplement once daily at 10.00 hours and AH silage with or without enzyme treatment was offered ad libitum to animals twice daily at 09.00 and 16.00 hours. Sheep were fed on four experimental forage diets comprised of AH silage and barley (D1), AH silage treated with ZAD1(®) and barley (D2), AH silage treated with ZAD2(®) and barley (D3) and AH silage treated with a combination of ZAD1(®) and ZAD2(®) (1:1) and barley (D4). Ensiling AH with enzymes reduced its contents of neutral detergent fiber and acid detergent fiber. The dry matter intake of AH of D2, D3 and D4 decreased (P < 0.001) as compared to D1. However, enzyme-treated diets had greater total digestible nutrients intake (P < 0.001) as compared to D1. The nutrients digestibility for D2, D3 and D4 were higher than those for D1 (P < 0.001), and were higher for D3 as compared to both D2 and D4. Sheep fed on D3 had highest (P < 0.001) ruminal total volatile fatty acids concentration, ammonia nitrogen concentration and microbial protein yield. It could be concluded that AH silage treated with ZAD1(®) or ZAD2(®) improved digestibility and rumen fermentation in sheep.

Effects of feeding dry or modified wet distillers grains with solubles with or without supplemental calcium oxide on ruminal metabolism and microbial enzymatic activity of beef cattle.

The objectives of this study were to determine the interaction of feeding dry (DDGS) or modified wet (MDGS) distillers grains with solubles (DGS) with or without supplemental CaO on in situ DM and NDF disappearance; ruminal pH, VFA, and methane concentration; and cellulase and xylanase activity. Fistulated steers (n = 8; average initial BW = 540 ± 250 kg) were used in a replicated 4 × 4 Latin square design. Treatments were arranged in a 2 × 2 factorial, and steers were randomly allotted to 1 of 4 dietary treatments: 1) 50% DDGS with 0% CaO, 2) 48.8% DDGS supplemented with 1.2% CaO, 3) 50% MDGS with 0% CaO, or 4) 48.8% MDGS supplemented with 1.2% CaO (DM basis). The remainder of the diet was husklage, dry-rolled corn, and vitamin and mineral supplement. There were no interactions (P ≥ 0.12) of DGS type and CaO addition on any parameters measured. Steers fed DDGS had a 17% increase (P < 0.01) in DMI compared to steers fed MDGS; however, CaO supplementation reduced (P = 0.03) DMI by 12%, regardless of DGS type. As expected, addition of CaO increased the pH of the diet by 1.82 pH units. This caused a time by CaO interaction (P = 0.05) for ruminal pH. Regardless of DGS type, steers supplemented with CaO tended to have increased (P = 0.09) ruminal pH at 1.5 h and had increased (P = 0.03) ruminal pH at 3 h postfeeding; however, ruminal pH did not differ (P ≥ 0.24) for the remainder of the day. There was no difference (P = 0.46) in ruminal cellulase activity when comparing type of DGS fed. However, there was a time by CaO interaction (P < 0.01); cattle fed 1.2% CaO diets had 28% greater ruminal cellulase activity only at 0 h postfeeding when compared to cattle fed 0% CaO. Furthermore, feeding supplemental CaO increased (P = 0.04) acetate to propionate ratio (A:P) regardless of type of DGS fed. Increased initial ruminal pH and cellulase activity from supplemental CaO did not increase (P = 0.48) in situ NDF disappearance. No differences (P ≥ 0.48) in ruminal methane concentration were found when comparing DGS type or supplemental CaO. In conclusion, the type of DGS fed had little effect on ruminal metabolism. Even though CaO increased ruminal pH and cellulase activity at some times postfeeding, it was not enough to affect in situ fiber disappearance.

Using a fibrolytic enzyme in barley-based diets containing wheat dried distillers grains with solubles: ruminal fermentation, digestibility, and growth performance of feedlot steers.

Two experiments were conducted to evaluate the effects of adding an exogenous fibrolytic enzyme (FE) on ruminal pH and fermentation, digestibility, and growth performance of feedlot beef cattle fed a finishing diet containing wheat dried distillers grains with solubles (DDGS). In Exp. 1, 4 ruminally cannulated Angus heifers (average BW of 807 ± 93.9 kg) were used in a replicated 4 × 4 Latin square design. Treatments were 1) control (CON; 10% barley silage and 90% barley grain-based concentrate), 2) CON diet substituting 30% wheat DDGS for barley grain (WDG), 3) WDG diet supplemented with low FE (WDGL), and 4) WDG diet supplemented with high FE (WDGH). Heifers fed WDG had less (P = 0.01) total tract DM digestibility than heifers fed CON. Increasing FE linearly (P < 0.05) increased starch digestibility without affecting digestibility of other nutrients. Addition of FE also reduced (P = 0.03) ruminal ammonia-N (NH3-N) concentration but did not affect VFA concentration. Moreover, application of FE to wheat DDGS linearly increased in situ ruminal DM (P < 0.01) and NDF (P = 0.02) disappearance after 48 h of incubation. In Exp. 2, 160 yearling steers (initial BW = 495 ± 37.9 kg) were fed the same diets as in Exp. 1. No differences in DMI, final BW, ADG, dietary NEg, or carcass characteristics were observed among diets. However, the steers fed WDG had less (P < 0.05) G:F and greater number of (P < 0.01) abscessed livers than steers fed CON. Increasing FE application in wheat DDGS diets did not affect DMI, final BW, or ADG but tended (P < 0.09) to linearly improve feed efficiency and decreased (P = 0.03) the incidence of abscessed livers. These results demonstrated adverse effects of including wheat DDGS in finishing diets on feed digestion, feed efficiency, and animal health. Application of FE in wheat DDGS-based diets potentially improved starch digestion, protein metabolism in the rumen, feed efficiency, and animal health.

Interactions between phytase and xylanase enzymes in male broiler chicks fed phosphorus-deficient diets from 1 to 18 days of age.

A total of 735 one-day-old male broiler chicks were used to evaluate the interactions between different levels of phytase and xylanase enzymes on performance and bone mineralization. Basal nonphytate P (nPP)-deficient diets (0.15%) were supplemented with different levels of phytase [0X, 1X, 2X, 3X, and 4X of recommended level (X = 500 phytase units per kg of feed)] alone or in combination with 3 levels of a xylanase preparation [0X, 1X, and 2X of recommended level (X = 0.1 g per kg of feed; a mixture with predominantly xylanase activity)]. A standard curve was developed using different levels of nPP (0.15 to 0.45%) to estimate the P equivalency of each enzyme combination. Treatments were replicated with 7 pens of 5 chicks each. The findings indicated that reducing dietary nPP level had a severely negative influence on bird performance and bone ash content. Results also showed that increasing levels of phytase significantly (P < 0.05) improved BW, feed intake, feed conversion ratio, mortality, and toe and tibia bone ash contents in a dose-dependent fashion. The P equivalency of phytase was also dose dependent, with estimates of 0.08, 0.11, 0.15, and 0.19 for 1X, 2X, 3X, and 4X supplementation levels of phytase, respectively. Xylanase preparation at 1X level failed to influence measured criterion; however, increasing the level of xylanase (2X) resulted in deteriorating BW and feed conversion ratio. The P equivalency of xylanase preparation at 1X and 2X was estimated as 0.010 and 0.014%. There were no interactions between phytase and xylanase preparation (P > 0.05). In conclusion, phytase was effective in improving bird performance and bone mineralization; however, xylanase supplementation failed to enhance phytase efficacy.

Interactive effects of phytase and xylanase supplementation with extractable salt-soluble protein content of corn in diets with adequate calcium and nonphytate phosphorus fed to broilers.

The objective was to determine the effects of extractable salt-soluble protein content of corn (PS) and exogenous enzyme supplementation on N, starch, and energy digestibility in broilers fed diets adequate in Ca and nonphytate P. Broilers were randomly distributed into floor pens (6 replicate pens per treatment) with 28 birds per pen at 1 d of age. Treatments consisting of 4 sources of corn varying in PS (A, 58.1; B, 54.2; C, 53.7; and D, 30.6 mg of BSA equivalent values) with or without phytase (0 and 1,000 phytase units/kg) and xylanase (0 and 16,000 units of xylanase activity/kg) were randomly assigned to each pen. Different sources of corn were provided from 1 to 9 and 24 to 29 d of age. However, enzyme treatments were provided throughout the experiment. From 1 to 9 d of age, no interactions were observed. Apparent ileal N digestibility (AIND) and apparent ileal digestible energy (IDE) of diets with the lowest PS (based on corn D) were lower (P ≤ 0.05) than those of diets with a higher PS. Phytase increased (P ≤ 0.01) AIND and IDE by 5 and 16%, respectively, and xylanase exerted the opposite effect (P ≤ 0.03). From 24 to 29 d of age, phytase and xylanase in combination resulted in reduced (P ≤ 0.05) AIND of diets with a low PS (based on corn D) compared with the basal diet in broilers. Broilers fed diets with the highest or lowest PS (based on corn A or D) had lower (3-way interaction; P ≤ 0.05) IDE when phytase and xylanase were supplemented in combination compared with either enzyme alone. In conclusion, responses to exogenous enzyme supplementation are not constant and are influenced by the source of ingredients as well as the age of broilers. The magnitudes of the responses to phytase on nutrient and energy digestibility were greater at 9 compared with 29 d of age.

Lipidemic responses of male broiler chickens to enzyme-supplemented wheat-soybean meal-based diets with various levels of metabolizable energy.

Effects of 2 various levels of AME (according to the manual recommendation and 100 kcal kg(-1) less than it), 2 levels of endo-beta-D-mannanase enzyme (0, 1 g kg(-1)) and 2 levels of xylanase enzyme (0 and 1 g kg(-1)) on serum lipid parameters as a 2(3) factorial arrangement were tested in 120 male broiler chicks fed wheat-soybean meal-based diet. These birds were randomly assigned to 8 experimental groups with 3 pen per group and 5 birds per pen. The serum HDL-cholesterol (HDL), LDL-cholesterol (LDL), Total-cholesterol (TC) and Triglycerides (TG) concentrations were measured at 31 and 41 day of age. The concentrations of serum TG, TC and LDL of 41-day-old birds demonstrated to be lower than those of 31-d-old (p < 0.001). Some hypolipidemic responses were observed in the broiler chicks fed on (1) Diet supplemented with only beta-mannanase, (2) Normal-AME diets supplemented with p-mannanase, (3) Normal-AME diets supplemented with Xylanse and (4) Normal-AME diets supplemented with both beta-mannanase and Xylanase (p < 0.01). In the other hand, some hyperlipidemic responses were detected in the broiler chicks fed on low-AME diets supplemented with xylanse or beta-mannanase enzymes, alone or in combination (p < 0.01). Regardless of AME, adding both xylanse and beta-mannanase to the wheat-soybean meal-based diets have both hyperlipidemic and hypolipidemic effects together (p < 0.01).