Exogenous Enzyme Product Research Articles

Feeding amylolytic and proteolytic exogenous enzymes: Effects on nutrient digestibility, ruminal fermentation, and performance in dairy cows

Exogenous enzymes are added to diets to improve nutrient utilization and feed efficiency. A study was conducted to evaluate the effects of dietary exogenous enzyme products with amylolytic (Amaize, Alltech) and proteolytic (Vegpro, Alltech) activity on performance, excretion of purine derivatives, and ruminal fermentation of dairy cows. A total of 24 Holstein cows, 4 of which were ruminally cannulated (161 ± 88 d in milk, 681 ± 96 body weight, and 35.2 ± 5.2 kg/d of milk yield), were blocked by milk yield, days in milk, and body weight, and then distributed in a replicated 4 × 4 Latin square design. Experimental periods lasted 21 d, of which the first 14 d were allowed for treatment adaptation and the last 7 d were used for data collection. Treatments were as follows: (1) control (CON) with no feed additives, (2) amylolytic enzyme product added at 0.5 g/kg diet dry matter (DM; AML), (3) amylolytic enzyme product at 0.5 g/kg of diet DM and proteolytic enzyme product at 0.2 g/kg of diet DM (low level; APL), and (4) amylolytic enzyme products added at 0.5 g/kg diet DM and proteolytic enzyme product at 0.4 g/kg of diet DM (high level; APH). Data were analyzed using the mixed procedure of SAS (version 9.4; SAS Institute Inc.). Differences between treatments were analyzed by orthogonal contrasts: CON versus all enzyme groups (ENZ); AML versus APL+APH; and APL versus APH. Dry matter intake was not affected by treatments. Sorting index for feed particles with size <4 mm was lower for ENZ group than for CON. Total-tract apparent digestibility of DM and nutrients (organic matter, starch, neutral detergent fiber, crude protein, and ether extract) were similar between CON and ENZ. Starch digestibility was greater in cows fed APL and APH treatments (86.3%) compared with those in the AML group (83.6%). Neutral detergent fiber digestibility was greater in APH cows compared with those in the APL group (58.1 and 55.2%, respectively). Ruminal pH and NH3-N concentration were not affected by treatments. Molar percentage of propionate tended to be greater in cows fed ENZ treatments than in those fed CON. Molar percentage of propionate was greater in cows fed AML than those fed the blends of amylase and protease (19.2 and 18.5%, respectively). Purine derivative excretions in urine and milk were similar in cows fed ENZ and CON. Uric acid excretion tended to be greater in cows consuming APL and APH than in those in the AML group. Serum urea N concentration tended to be greater in cows fed ENZ than in those fed CON. Milk yield was greater in cows fed ENZ treatments compared with CON (32.0, 33.1, 33.1, and 33.3 kg/d for CON, AML, APL, and APH, respectively). Fat-corrected milk and lactose yields were higher when feeding ENZ. Feed efficiency tended to be greater in cows fed ENZ than in those fed CON. Feeding ENZ benefited cows' performance, whereas the effects on nutrient digestibility were more pronounced when the combination of amylase and protease was fed at the highest dose.

Effect of Exogenous Enzyme Mixture on Growth Performance, Digestibility and Some Rumen Parameters of Finishing Lambs Fed High Whole Grain Hulled Barley Diet

Background: Whole grain hulled barley has several nutritional limitations in diets for ruminants. Its fibrous hull which is resistant to ruminal microbial degradation and this could decrease performance. Therefore, tools that optimize the use of barley hull are required. Exogenous enzyme products including fibrolytic enzymes may be an important tool to improve the digestibility of whole grain, feed and improve the efficiency of lambs. Thus, this study aimed to investigate the effects of the supplementary exogenous enzyme mixture at three different levels on growth performance, digestibility and ruminal fermentation of male finishing lambs fed diets containing whole barley grain. Methods: Twenty-one Karayaka lamb (29.36±1.32 kg initial weight) were assigned to three experimental treatments: (1) Control, diet without exogenous enzyme; (2) 5 exogenous enzyme, diet with 5 g exogenous enzyme/head/day) and (3) 10 exogenous enzyme, diet with 10 exogenous enzyme g/head/day). Forage-free finishing diets were composed of 70% whole barley and 30% protein supplement. The experimental diet was offered to the animals for 56 days. Nutrient digestibility was determined during the last 4 days of the experiment. Result: The supplementation of exogenous enzyme did not affect dry matter intake. Increasing exogenous enzyme in whole barley grain-based diets did not alter digestibility of dry matter, organic matter, crude protein and neutral detergent fibre. Regarding, mean ruminal pH, NH3-N and ruminal total volatile fatty acid, there was no difference between control and both exogenous enzyme mixture levels. The findings of the current study indicate that there was no effect of exogenous enzyme mixture on lamb performance, digestibility and ruminal fermentation fed whole grain hulled barley-based diets in Karayaka lambs. Therefore, supplementing such diets with exogenous enzyme for fatting Karayaka lambs is not recommended.

Characterization, screening, and application of bacteria with probiotic properties isolated from the gut of Labeo calbasu (Hamilton)

Abstract Labeo calbasu is not reared in aquaculture because of the unavailability of proper feed and the risk of infection. The ability of probiotics to promote growth and immunological defense barriers in the gut has led to the use of autochthonous probiotic bacteria. Hence, the present study was conducted to screen potential probiotic bacteria from the gut of L. calbasu and to supplement the same to study the effect on growth and immunity. Strains with probiotic properties were assayed for exogenous enzyme production and identified based on phylogenetic analysis and nucleotide homology. Four of nine isolates showed promising antibacterial activity against Aeromonas hydrophila. The strains survived in relatively low pH and high bile concentrations and were investigated for hydrophobicity, autoaggregation, and coaggregation. Based on the positive results of this study, the LC1 and LC4 strains (analysed with 16S rRNA gene sequencing and showing similarity with Aneurinibacillus aneurinilyticus and Stenotrophomonas sp.) were further incorporated into diets to explore their probiotic effects on the growth performance and immunity of the fish. Administering the probiotics elevated growth parameters and benefitted the immune system by increasing the phagocytic index and decreasing the mortality rate with exposure to A. hydrophila.

Effects of Saccharomyces cerevisiae-based direct-fed microbial and exogenous enzyme products on enteric methane emission and productivity in lactating dairy cows

The objective of this experiment was to investigate the effects of a Saccharomyces cerevisiae-based direct-fed microbial product (SDM) and an exogenous enzyme product (ENZ) on enteric methane emission, milk yield and composition, total-tract digestibility of nutrients, ruminal fermentation, and nitrogen excretion and secretion in lactating dairy cows. Eighteen Holstein cows were used in a 3 × 3 Latin square design experiment with three 28-d periods. Treatments were (1) control (no additive), (2) 28 g of SDM/d per cow, or (3) 10 g of ENZ/d per cow. Treatments were top-dressed at the time of feeding. The basal diet consisted of (dry matter basis) 60% forage and 40% concentrates and contained 16.5% crude protein and 32.0% neutral detergent fiber. Treatments had no effect on enteric methane production, yield (methane per kg of dry matter intake, DMI), or intensity (methane per kg of energy-corrected milk yield). Carbon dioxide production was similar among treatments. Compared with control, SDM increased milk yield by 2 kg/d without affecting DMI or feed efficiency. Supplementation of the diet with ENZ did not affect DMI, milk yield, or feed efficiency. Concentrations and yields of milk fat, true protein, and lactose, and energy-corrected milk yield were not different among treatments. Neither SDM nor ENZ had an effect on total-tract digestibility of nutrients or nitrogen excretion and secretion. Concentration of total volatile fatty acids (VFA) in ruminal fluid was increased by both SDM and ENZ, and rumen pH was decreased by SDM compared with the control. At levels similar to the control DMI, the increased concentration of VFA in ruminal fluid of cows receiving SDM suggests an increased postruminal supply of energy and may partly explain the increased milk yield with that treatment. However, it is important to note that milk composition and energy-corrected milk yield were not affected by treatment.

Analysis of the mould microbiome and exogenous enzyme production in Moutai-flavor Daqu

The objective of this study was to isolate and screen the mould microbiome of five Moutai-flavor Daqu samples and to characterize and quantify the associated exogenous enzymes. In all, 11 and 18 mould genera were identified in the samples by, respectively, a culture-dependent method and a culture-independent high throughput sequencing method. Species of Aspergillus and Rhizopus were the dominant mould genera in the Moutai-flavor Daqu samples. Analysis of enzyme production showed that Aspergillus versicolor in Daqu ZX01 exhibited the highest neutral proteinase activity (2671.0 ± 73.9 U/g) and Rhizomucor pusillus in Daqu DYT01 exhibited the highest saccharifying amylase production ability (1724.6 ± 11.0 U/g). Copyright © 2017 The Institute of Brewing & Distilling

Effects of an enzyme complex on in vitro dry matter digestibility of feed ingredients for pigs.

Feed ingredients of plant origin are commonly used in swine diets. However, the major components of plant cell walls, non-starch polysaccharides (NSPs), reduce nutrient digestibility. To improve the efficiency of feed utilization, exogenous enzyme products that degrade NSPs have been widely used in commercial animal feeds. Nonetheless, the effects of exogenous enzyme addition to swine diets on nutrient digestibility have not been determined. To this end, in vitro approaches may be used. The objective of this study was to determine the effects of an enzyme complex (EC) containing xylanase, protease, and phytase on the in vitro dry matter (DM) digestibility of nine feed ingredients including cereal grain energy sources (corn, wheat, and barley) and protein sources (soybean meal, rapeseed meal, palm kernel meal, cottonseed meal, copra meal, and distillers dried grains with solubles). Both in vitro ileal and total tract digestibility (IVID and IVTTD, respectively) of DM were determined for the nine test ingredients, with or without EC addition. The EC addition increased the IVID of DM in copra meal (p = 0.047) and tended to increase the IVID of DM in corn, wheat, barley, palm kernel meal, cottonseed meal, and DDGS (p < 0.10). On the other hand, no significant effect was observed in soybean meal and rapeseed meal. The IVTTD of DM in the test ingredients was not affected by the addition of EC, except for cottonseed meal (52.1 vs. 50.6%, p = 0.053). In conclusion, the effects of EC addition on in vitro DM digestibility may vary, depending on the test ingredient and method used.

Effects of exogenous enzymes on nutrient digestibility, ruminal fermentation and growth performance in beef steers

Forty crossbred steers (Baladi×Friesian, average BW 153±5.14kg) were used to evaluate the effects of exogenous enzyme (ENZ) addition on nutrient intake, digestion, ruminal fermentation and feed conversion in beef steers. Steers were randomly assigned to two groups of 20 animals and fed individually a total mixed ration (TMR) without (CTRL) or with addition of 40g/hd/d of an enzyme mixture (ZADO®). The ENZ mixture was added for 220 days and in vivo apparent digestibility was measured on days 210–220. Enzyme addition did not affect (P=0.1) DM intake, whereas it increased (P<0.05) total tract apparent digestibility of all nutrients. The magnitude of improvement in digestibility varied among nutrients, with the highest improvement occurring in digestibility of NDF and ADF (21.8% and 26.7%, respectively). Addition of ENZ also increased (P<0.05) concentrations of rumen ammonia N and total short chain fatty acids (SCFA) before and 3h post-feeding. Allantoin concentration total purine derivates were increased (P=0.04) with enzyme addition while uric acid was not affected (P=0.05). Live-weight gain was also higher (P<0.01) in steers supplemented with ENZ. In conclusion, adding the exogenous enzyme product increased live-weight gain by 16% due to increased nutrient digestibility.

Influence of exogenous enzymes on nutrient digestibility, extent of ruminal fermentation as well as milk production and composition in dairy cows

This experiment studied effects of a mixture of exogenous enzymes (ZADO ®) from anaerobic bacteria on ruminal fermentation, feed intake, digestibility, as well as milk production and composition in cows fed total mixed rations (TMRs; 0.7 corn silage and 0.3 of a concentrate mixture). Twenty lactating multiparous Brown Swiss cows (500 ± 12.4 kg live weight) were randomly assigned into two experimental groups of 10 immediately after calving and fed a TMR with or without (CTRL) addition of 40 g/cow/d of enzymes for 12 weeks. Addition of enzymes increased ( P<0.05) rumen microbial N synthesis. Intake of dry matter (DM) and organic matter (OM) was positively influenced ( P<0.05) by supplementation, and digestibility of all nutrients was higher ( P<0.05) in the total tract of supplemented cows, although the magnitude of the improvement varied among nutrients, with the highest improvement in aNDFom and ADFom (418–584 and 401–532 g/kg respectively; P<0.05) than the other nutrients. Supplementation of enzymes also increased ( P<0.05) rumen ammonia N and total short chain fatty acid (SCFA) concentrations, and individual SCFA proportions were also altered with an increase in acetate (61.0–64.8 mol/100 mol; P=0.05) before feeding, and acetate and propionate increased 3 h post-feeding (60.0–64.0 and 18.3–20.8 mol/100 mol respectively; P<0.05). Milk and milk protein production was higher (12.8–15.7 and 0.45–0.57 kg/d respectively; P<0.05) for cows fed the ZADO ® supplemented diet. This exogenous enzyme product, supplemented daily to the TMR of cows in early lactation, increased milk production due to positive effects on nutrient intake and digestibility, extent of ruminal fermentation and microbial protein synthesis.

A comparison of HPLC and spectrophotometrical methods to determine the activity of ferulic acid esterase in commercial enzyme products and rumen contents of steers

The aim of this study was to compare different methods to determine ferulic acid esterase (FAE) activity in commercial enzyme products and to measure the FAE activity in the rumen of steers fed different diets (A: 5 kg forage and 4 kg concentrate; B: 5 kg forage and 5 kg concentrate; C: 5 kg forage and 6 kg concentrate). FAE activities in seven commercial enzyme products were determined by high performance liquid chromatography (HPLC) at a wavelength of 320 nm and by spectrophotometric assays at wavelengths of 335 and 340 nm. FAE activities varied dramatically among the seven exogenous enzyme preparations. Correlation between the enzyme activities determined by HPLC and spectrophotometric assays was high ( r > 0.98), but consistency among the different methods was poor, especially with the spectrophotometrical method (SP) at 335 nm. Therefore, the SP by microplate reader at 340 nm with standard substrate of methyl ferulate (MFA) was recommended for rapid analysis of large numbers of samples. Rumen fluid of steers had low, and different, ferulic acid esterase activity at different times of the day. Furthermore, high diurnal variation occurred for the ferulic acid esterase and other fibrolytic enzymes that were measured. The activity of FAE and other fibrolytic enzymes differed along with the diet compositions and feed intake. Diet C had the highest mean fibrolytic enzyme activity in the rumen, and diet A had the lowest activity of FAE and birchwood xylanase (BX). High correlations occurred between activities of carboxymethyl cellulase (CMC), BX, acetyl esterase (AE) and those of FAE, both with all data and within each diet. As an accessory enzyme, ferulic acid esterase should not be neglected in studies on furthering understanding of biodegradation of plant cell wall in rumen, and impacts of exogenous enzyme products.

ENZIMAS EXÓGENAS NA ALIMENTAÇÃO DE AVES

Currently the great quandary in the animal production is the reduction of the costs, without forgetting the maintenance them profits or until the increase of them. Other aspect sufficiently questioned in the present time is foods alternative that they are used in the substitution of the maize and soy, that I eat known for, suffers many influences of market and its price varies very. However the great majority of foods used in this substitution contains antinutricionais factors as inhibiting of proteases, fitatos among others that they get worse the quality of the food and makes it difficult the digestibilidade of the nutrients on the part of the birds. In search of this it is that if it has launched hand of the technology in vigor in the exogenous enzyme production for the animal feeding, having as benefits the maintenance of the quality of foods, improves in the digestibilidade of the nutrients, reduction of the humidity of excrements and elimination wants either in part or total of the antinutricionais factors of foods used in the feeding of birds. Being thus this revision it had as objective to elaborate a revision on the basis of the advances and too much uses of exogenous enzymes in the feeding of birds.

Assessment of the efficacy of varying experimental exogenous fibrolytic enzymes using in vitro fermentation characteristics

This study evaluated a series of recombinant, single activity experimental enzyme products including 13 endoglucanases (END) and 10 xylanases (XY), for their potential to improve in vitro ruminal degradation of alfalfa hay in two experiments. Based on the endoglucanase or xylanase enzymatic activities measured using complex substrates at the optimal conditions (pH 5.4, 37 °C) for the enzymes, a dose level (1 unit/g dry matter [DM]) was chosen for addition of enzymes to substrate. Enzyme products, re-suspended with water, were added to alfalfa hay (0.5 or 1.0 g DM) in culture vials in six replications. Anaerobic buffer medium (20 or 40 ml) adjusted to pH 6.0 and strained ruminal fluid (5 or 10 ml) were sequentially added to the vials and incubated for 18 h. Headspace gas production (GP) was measured throughout the incubation, and degradability of organic matter (OMD) and fibre and volatile fatty acid (VFA) concentrations were determined after 18 h of incubation. The enzyme products had a wide range of added endoglucanase or xylanase activities when determined using pure substrates and physiological conditions typical of the rumen (pH 6.0, 39 °C). In experiment 1, many END, and some XY, products increased GP and OMD. The correlation between added endoglucanase activity determined at ruminal conditions and OMD improvement was high ( r = 0.71; P<0.01), whereas added activity of xylanase was not associated with OMD improvement. Two END and two XY products selected from experiment 1 were further assessed because they substantially improved GP and OMD. In experiment 2, all enzyme treatments, alone or in combination, increased total GP and DM and fibre degradabilities (P<0.05). However, the combinations of END and XY did not increase degradation of alfalfa beyond that of the component enzymes. Total VFA production was not affected by enzyme treatments although some products changed the acetate to propionate ratio. Experimental exogenous enzyme products with either endoglucanase or xylanase activity substantially improved in vitro ruminal degradation of alfalfa hay, but further improvement by combining these activities did not occur.

Effect of food enzymes on AME and composition of digesta from broiler chickens fed on diets containing non-starch polysaccharides isolated from lupin kernel

1. The effects of lupin non-starch polysaccharides (NSPs) on apparent metabolisable energy (AME) and composition of digesta from broiler chickens were examined in a dose-response experiment in conjunction with a single combination of 2 commercial enzyme products known from previous studies to partially depolymerise NSPs in diets containing sorghum, casein and lupin kernel. The 7-d experiment was conducted on broiler chickens (24 to 31 d of age) in metabolism cages. 2. The basal diet used in this study contained (in g/kg) sorghum (low tannin) 800, casein 134, celite 20 and vitamins and minerals 46. The inclusion rate for each enzyme product was 0.2 g/kg. Lupin kernel isolate (containing 560 g/kg total NSPs) replaced some of the sorghum and casein. The dose rates of lupin kernel isolate (50, 100 and 150 g/kg) were equivalent to dietary inclusion rates of 80, 160 and 240 g/kg, approximately, of untreated lupin kernel from Lupinus angustifolius cultivar Gungurru. 3. Inclusion of lupin kernel isolate in the diet depressed AME and bird performance, and raised ileal viscosity and excreta moisture, in a dose-dependent manner. 4. A combination of 2 proprietary enzyme products improved dietary AME by 0.3 MJ/kg dry matter. Enzymes reduced excreta moisture when inclusion of lupin kernel isolate was 50 g/kg (equivalent to lupin kernel at 80 g/kg, approximately) but not at higher inclusions. 5. Food enzymes partially depolymerised some insoluble NSPs in lupin kernel isolate, which increased ileal viscosity and soluble NSP content of digesta. 6. It was concluded that development of appropriate exogenous enzyme products could lead to lupin NSPs being used as an energy source for poultry.