Addition Of Exogenous Enzymes Research Articles

PSII-14 Economic impact of adding exogenous enzymes in a sorghum-based diet for growing and finishing pigs

Abstract Food efficiency in production aims for better productive results in body weight gain and reduction of dietary costs. Sorghum is a great cereal to use in pig feed, but it has a greater concentration of phytate than corn, which is not degraded by monogastric animals. The addition of exogenous enzymes aims to increase nutrient digestibility, providing greater efficiency of use. The study aim was to evaluate the performance of the animals and the economic viability of the joint use of protease and phytase in sorghum-based diets of sorghum for finishing pigs. The experiment was carried out on a commercial farm, in Patos de Minas, Minas Gerais, Brazil. Commercial hybrid pigs (n = 240; 70 d old; body weight = 26 kg) were used. Individually identified and distributed in pens respecting body weight and sex blocks in a randomized block design, two treatments with 12 replications each were evaluated, totaling 24 experimental plots, with 10 animals each. In treatment 1, control diet based on sorghum and soybean meal (SBM) with phytase 600 FYT/kg of feed (HiPhorius); and treatment 2, same composition as treatment 1 with addition of 30,000 NFP/kg of feed of protease (ProAct 360). The diets were formulated according to Brazilian tables and use nutritional matrix of enzymes (phytase and protease), excepted in control group. At 0, 48, 70 and 89 d of the experiment, the animals were weighed individually, and all the food provided was weighed and the leftovers were quantified, providing information for calculating the variables: average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR). The feeding program costs were calculated. The results were submitted to the variance test, using the statistical program SAS, version 9.4. The difference between mean values was considered statistically significant when P < 0.050, using the Turkey test. There was no statistical difference (P > 0.05) between treatments in relation to ADFI between 0 to 49 d (P = 0.110), 0 to70 d (P = 0.431) and 0 to 89 d of experiment (P = 0.490). There was also no impact on ADG at 0 to 49 d (P = 0.325), 0 to 70 d (P = 0.834) and 0 to 89 experimental days (P = 0.915); and FCR at 0 to 49 days (P = 0.304), 0 to 70 d (P = 0.470) and 0 to 89 experimental days (P = 0.177). Although, there was no effect on the performance indices evaluated in this study, the use of exogenous enzymes such as proteases can have an economic impact on the composition of the pig diet, as it reduced the nutritional cost, increased the revenue per pig produced, increased the margin on feed in 1.06% and reducing the cost per kilogram produced by 1.18%. Therefore, the association of protease and phytase showed a significant economic impact when used in diets based on sorghum and SBM for growing and finishing pigs, even with no direct impact on performance indices.

Exogenous enzyme addition affects litter decomposition by altering the microbial community and litter nutrient content in planted forest

Abstract Litter inputs have great impacts on the soil properties and ecosystem functioning in forests. Rapid litter decomposition leads to decreases in planted forest agricultural waste and enhances the nutrient cycle in forests. The breakdown of litter and the release of various components depend heavily on enzymes. However, the effects of exogenous enzyme preparations on litter decomposition have been hardly investigated. In this study, we examined how these enzymes affected the remaining rate of litter quality, nutrient content (C, N, K), and microbial community diversity. Taking Eriobotrya japonica litter as the research object, five exogenous enzymes (laccase, lignin peroxidase, leucine arylamidase, cellulase, and acid phosphatase) were applied to litter leaves. The mass remaining rate and main nutrient content of the litter were measured during the decomposition period. The microbial diversity attached to the surface of the litter was determined after decomposition at constant temperature and humidity for 189 days. Application of laccase and lignin peroxidase increased litter degradation by affecting microbial diversity, N and K contents. Addition of leucine arylamidase leaded to an increase in N content, and decreased the quality of the litter. The cellulose and lignin decomposition rate in litters was unaffected by the addition of cellulase, laccase, and lignin peroxidase. These results indicate that exogenous addition of enzymes may alter the nutrient content and microbial community, thus affecting litter decomposition. It is imperative to investigate the effects and mechanisms of exogenous enzymes on litter decomposition for regulating decomposition of agricultural waste litter.

Engineering natural isolates of Saccharomyces cerevisiae for consolidated bioprocessing of cellulosic feedstocks

Saccharomyces cerevisiae has gained much attention as a potential host for cellulosic bioethanol production using consolidated bioprocessing (CBP) methodologies, due to its high-ethanol-producing titres, heterologous protein production capabilities, and tolerance to various industry-relevant stresses. Since the secretion levels of heterologous proteins are generally low in domesticated strains of S. cerevisiae, natural isolates may offer a more diverse genetic background for improved heterologous protein secretion, while also displaying greater robustness to process stresses. In this study, the potential of natural and industrial S. cerevisiae strains to secrete a core set of cellulases (CBH1, CBH2, EG2, and BGL1), encoded by genes integrated using CRISPR/Cas9 tools, was evaluated. High levels of heterologous protein production were associated with a reduced maximal growth rate and with slight changes in overall strain robustness, compared to the parental strains. The natural isolate derivatives YI13_BECC and YI59_BECC displayed superior secretion capacity for the heterologous cellulases at high incubation temperature and in the presence of acetic acid, respectively, compared to the reference industrial strain MH1000_BECC. These strains also exhibited multi-tolerance to several fermentation-associated and secretion stresses. Cultivation of the strains on crystalline cellulose in oxygen-limited conditions yielded ethanol concentrations in the range of 4–4.5 g/L, representing 35–40% of the theoretical maximum ethanol yield after 120 h, without the addition of exogenous enzymes. This study therefore highlights the potential of these natural isolates to be used as chassis organisms in CBP bioethanol production.Key points• Process-related fermentation stresses influence heterologous protein production.• Transformants produced up to 4.5 g/L ethanol, ~ 40% of the theoretical yield in CBP.• CRISPR/Cas9 was feasible for integrating genes in natural S. cerevisiae isolates.

Catalytic Antibody Blunts Carfentanil-Induced Respiratory Depression.

Carfentanil, the most potent of the fentanyl analogues, is at the forefront of synthetic opioid-related deaths, second to fentanyl. Moreover, the administration of the opioid receptor antagonist naloxone has proven inadequate for an increasing number of opioid-related conditions, often requiring higher/additional doses to be effective, as such interest in alternative strategies to combat more potent synthetic opioids has intensified. Increasing drug metabolism would be one strategy to detoxify carfentanil; however, carfentanil's major metabolic pathways involve N-dealkylation or monohydroxylation, which do not lend themselves readily to exogenous enzyme addition. Herein, we report, to our knowledge, the first demonstration that carfentanil's methyl ester when hydrolyzed to its acid was found to be 40,000 times less potent than carfentanil in activating the μ-opioid receptor. Physiological consequences of carfentanil and its acid were also examined through plethysmography, and carfentanil's acid was found to be incapable of inducing respiratory depression. Based upon this information, a hapten was chemically synthesized and immunized, allowing the generation of antibodies that were screened for carfentanil ester hydrolysis. From the screening campaign, three antibodies were found to accelerate the hydrolysis of carfentanil's methyl ester. From this series of catalytic antibodies, the most active underwent extensive kinetic analysis, allowing us to postulate its mechanism of hydrolysis against this synthetic opioid. In the context of potential clinical applications, the antibody, when passively administered, was able to reduce respiratory depression induced by carfentanil. The data presented supports further development of antibody catalysis as a biologic strategy to complement carfentanil overdose reversal.

The role of lignin in the conversion of wheat straw to cellobionic acid by Neurospora crassa HL10

An engineered Neurospora crassa strain (HL10) naturally produces cellulases, cellobiose dehydrogenase (CDH), and heterologously expresses laccase. It can convert Avicel (cellulose) to cellobionic acid (cellobionate) without any enzyme addition at high yields. However, the addition of a catalytic amount of artificial redox mediators was required to achieve a high yield conversion. When a lignocellulosic substrate like wheat straw was used as the carbon source, it was found that adding exogenous artificial redox mediators did not improve cellobionate yield. We hypothesized that lignin and lignin degradation products were able to serve as redox mediators for the CDH-laccase conversion system. Experiments were designed to prove the hypothesis. When N. crassa HL10 was grown on Avicel, the addition of enzyme hydrolyzed lignin (EHL) can achieve a similar level of cellobionate production as with ABTS addition. The addition of EHL can increase the cellobionate production rate in the CDH-laccase bi-enzyme system using cellobiose as the substrate. The formation of lignin radicals, and the quenching of lignin radicals by reduced CDH, were verified by an electron paramagnetic resonance (EPR) experiment, providing further evidence that lignin radicals can serve as the electron acceptor of reduced CDH. Hence, lignocellulosic biomass is a self-sufficient substrate for the production of cellobionate. The conversion of the lignocellulosic substrate to cellobionate using N. crassa HL10 can be achieved without any exogenous enzyme addition or redox mediator addition.

Enzymatic treatment in black tea manufacturing processing: Impact on bioactive compounds, quality, and bioactivities of black tea

This study investigated the changes of main quality components and the effect of bioactivities of black tea during a manufacturing process that adds to enzymatic treatment. It used black tea harvests in summer and autumn for manufacturing and analyzed the number of main quality components and bioactivities of the black tea produced by treating cellulase during rolling and polyphenol oxidase (PPO) and peroxidase (POD) during fermentation. The results found that the contents of total catechins (TC), total polyphenols (TP), reducing sugars (ReS), and individual catechins in the tea infusions of summer and autumn black tea that were obtained during enzymatic treatments decreased significantly (p < 0.05). However, the content of free amino acid (FAA), the tea quality parameter such as the total liquor color (TLC), the briskness index (BI), and the tea pigments including total theaflavin (TF), thearubigens (TR), TRSI, and TRSII all increased significantly (p < 0.05). Moreover, the biological activity enhanced black tea's antioxidant and metabolic capacity with enzymatic treatments were proved by in vitro experiments. The results of this study contribute to gaining further knowledge on how black tea's quality and potential health benefits are maximized by using exogenous enzyme addition methods in the manufacturing process preparation.

Combined effects of dietary Laminaria digitata with alginate lyase on plasma metabolites and hepatic lipid, pigment and mineral composition of broilers

BackgroundThe Laminaria digitata is an abundant macroalga and a sustainable feedstock for poultry nutrition. L. digitata is a good source of essential amino acids, carbohydrates and vitamins, including A, D, E, and K, as well as triacylglycerols and minerals, in particular iron and calcium. However, the few studies available in the literature with broilers document the application of this macroalga as a dietary supplement rather than a feed ingredient. No study has addressed up until now the effects of a high-level incorporation (> 2% in the diet) of L. digitata on plasma biochemical markers and hepatic lipid composition, as well as minerals and pigments profile in the liver of broilers. Our experimental design included one hundred and twenty Ross 308 male birds contained in 40 wired-floor cages and distributed to the following diets at 22 days of age (n = 10) for 15 days: 1) a corn-soybean basal diet (Control); 2) the basal diet plus 15% of L. digitata (LA); 3) the basal diet plus 15% of L. digitata with 0.005% of Rovabio® Excel AP (LAR); and 4) the basal diet plus 15% of L. digitata with 0.01% of the recombinant CAZyme, alginate lyase (LAE).ResultsL. digitata compromised birds’ growth performance by causing a reduction in final body weight. It was found an increase in hepatic n-3 and n-6 fatty acids, in particular C18:2n-6, C18:3n-6, C20:4n-6, C20:5n-3, C22:5n-3 and C22:6n-3 with the addition of the macroalga, with or without feed enzymes, to the broiler diets. Also, the beneficial C18:3n-3 fatty acid was increased by combining L. digitata and commercial Rovabio® Excel AP compared to the control diet. The sum of SFA, MUFA and the n-6/n-3 PUFA ratio were decreased by L. digitata, regardless the addition of exogenous enzymes. β-carotene was enhanced by L. digitata, individually or combined with CAZymes, being also responsible for a positive increase in total pigments. Macrominerals, in particular phosphorous and sulphur, were increased in the liver of broilers fed L. digitata individually relative to the control. For microminerals, copper, iron and the correspondent sum were consistently elevated in the liver of broilers fed L. digitata, individually or combined with exogenous CAZymes. The powerful discriminant analysis tool based on the hepatic characterization revealed a good separation between the control group and L. digitata diets but failed to discriminate the addition of feed enzymes.ConclusionsOverall, this study highlights the value of L. digitata as a feed ingredient for the poultry industry. Moreover, we can conclude that the effect of L. digitata overpowers the effect of feed enzymes, both the Rovabio® Excel AP and the alginate lyase. Having in mind the negative effects observed on birds’ performance, our main recommendation at this stage is to restraint L. digitata incorporation level in forthcoming nutritional studies.

Dehydrated citrus pulp reduces plasma cholesterol of weaned piglets, and an exogenous enzyme complex improves plasma calcium and performance

Dehydrated citrus pulp (DHCP) is a by-product from the orange industry available for use in diets for pigs in tropical countries; however, the high fiber content may limit its use for piglets, a fact that can be mitigated through the use of exogenous enzymes. The objective of this work was to evaluate the inclusion of DHCP with or without an exogenous enzyme complex in the diet of weaned piglets from 7 to 11 kg on performance, economic viability, and biochemical, hematological, and fecal parameters. Sixty-four crossbred piglets were used (32 barrows and 32 gilts) with an average initial weight of 6.52 ± 0.22 kg, weaned at 21 days of age and housed in pairs (one barrow and one gilt). Animals were distributed in a randomized block design, in a 2 × 2 factorial scheme. The treatments evaluated were the addition or absence of DHCP (30 g/kg) X the addition or absence of the exogenous enzyme complex to the diets, with eight replicates per treatment and two animals per experimental unit. The exogenous enzyme complex was a thermo-stable powder containing 2,700 U/g xylanase, 1,520 U/g β-glucanase, 1,650 U/g alpha-galactosidase, and 1,800 U/g cellulase (sourced by Aspergillus niger and expressed in Trichoderma reesei), plus 5,000 U/g phytase (sourced by Buttiauxella spp. and expressed in Trichoderma reesei). It was added to the diets according to the manufacturer's recommendation (130 g/ton of feed). There was no interaction (P > 0.05) between DHCP and exogenous enzymees for any variable. Even so, the use of DHCP reduced (P = 0.018) the total cholesterol concentration, from 96.67 to 88.29 mg/dL. There was an effect (P = 0.018) of exogenous enzymes on the final weight, with average values being 11.084 and 11.439 kg for animals fed diets without or with the addition of exogenous enzyme complex, respectively. The addition of exogenous enzymes increased the concentration of circulating calcium (P = 0.033), with average results of 7.03 and 8.49 mg/dL for animals fed without or with exogenous enzymes in the feed, respectively. The inclusion of 30 g/kg of DHCP in a piglet's diet (7 to 11 kg) reduces cholesterol concentration in its plasma, and the inclusion of exogenous enzymes provides greater final weight, economic return, and calcium concentration in plasma.

Early growing performance of local male weaning lambs fed edamame pod-based rations with and without exogenous enzyme additives

This study aimed to evaluate the early growing performance of local male weaning lambs fed edamame pod-based rations with and without exogenous enzyme additives. Nine local male weaning lambs with initial weight about 16.88±2.33 kg/head were randomly assigned into into 3 levels treatment using an experimental completely randomized design (CRD). Every single level treatment was repeated 3 times. The levels treatment was 0% (control), 0.14% (w/w), and 0.28% (w/w) of exogenous enzyme addition into edamame pod-based rations. The lambs were maintained in individual pen which fed and given drinking water ad libitum. The parameters observed were nutrients consumption, liveweight gain, and gain to feed ratio during the first couple week maintenance with 2 weeks adaptation period. Data obtained were analysis using analysis of variance and further tested by DMRT for different result. The result presented that nutrient consumption, daily gain and gain to feed ratio were not affected (P>0.05) by various levels of exogenous enzyme additions. Thereby, it was concluded that the addition of exogenous enzyme up to 0.28% w/w potentially improved the nutrient consumptions and productivity performance of local male weaning lambs.

Grape pomace in equine nutrition: effect on antioxidant status

Grape pomace in equine nutrition: effect on antioxidant status

Evaluation of Brewers Spent Grains with different levels of exogenous enzymes on the production performance and body composition of Nile tilapia (Oreochromis niloticus) and channel catfish (Ictalurus punctatus)

ABSTRACT This study evaluated Brewers Spent Grains (BSG) and an enzyme additive (AllzymeTM; ENZ) in practical diets for juvenile (~5 g/fish) Nile tilapia (Trial 1) and channel catfish (Trial 2). Following a factorial arrangement, effects of three dietary levels of BSG (0, 270, and 550 g/kg) and three levels of ENZ (0, 0.2, and 0.4 g/kg) on production performance and whole-body composition of the fish were evaluated. Upon conclusion of the feeding trials, no effects of ENZ, or interactions between BSG and ENZ, were found. Fish survival was >85% and unaffected by diet. Reduced growth of tilapia (from 949% to 778%) and catfish (from 1802% to 1364%) as dietary BSG increased from 0 to 550 g/kg were observed. In these trials, weight gain was reduced in both channel catfish and tilapia at BSG inclusion rates of ≥270 g/kg and that the addition of exogenous enzymes did not improve BSG utilization.

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

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

Improvement of nutrient bioavailability in millets: Emphasis on the application of enzymes.

Millets are a traditional staple food of the dryland regions of the world and are rich in essential nutrients like protein, fatty acids, minerals, vitamins, and dietary fiber. Also, millets commonly synthesize a range of secondary metabolites to protect themselves against adverse conditions. These factors are collectively termed anti-nutritional factors and the existence of these factors in millets might reduce the accessibility of the nutrients in humans. Some of these factors include protease inhibitors, tannins, non-starch polysaccharides-glucans, phytates, and oxalates each of which might directly or indirectly affect the digestibility of nutrients. Methods like soaking, germination, autoclaving, debranning, and the addition of exogenous enzymes have been used to reduce the anti-nutritional factors and elevate the bioavailability of the nutrients. This review summarizes various methods that have been used to improve nutrient bioavailability, specifically emphasizing the use of enzymes to improve nutrient bioavailability from millets. © 2021 Society of Chemical Industry.

Coculture with hemicellulose-fermenting microbes reverses inhibition of corn fiber solubilization by Clostridium thermocellum at elevated solids loadings

BackgroundThe cellulolytic thermophile Clostridium thermocellum is an important biocatalyst due to its ability to solubilize lignocellulosic feedstocks without the need for pretreatment or exogenous enzyme addition. At low concentrations of substrate, C. thermocellum can solubilize corn fiber > 95% in 5 days, but solubilization declines markedly at substrate concentrations higher than 20 g/L. This differs for model cellulose like Avicel, on which the maximum solubilization rate increases in proportion to substrate concentration. The goal of this study was to examine fermentation at increasing corn fiber concentrations and investigate possible reasons for declining performance.ResultsThe rate of growth of C. thermocellum on corn fiber, inferred from CipA scaffoldin levels measured by LC–MS/MS, showed very little increase with increasing solids loading. To test for inhibition, we evaluated the effects of spent broth on growth and cellulase activity. The liquids remaining after corn fiber fermentation were found to be strongly inhibitory to growth on cellobiose, a substrate that does not require cellulose hydrolysis. Additionally, the hydrolytic activity of C. thermocellum cellulase was also reduced to less-than half by adding spent broth. Noting that > 15 g/L hemicellulose oligosaccharides accumulated in the spent broth of a 40 g/L corn fiber fermentation, we tested the effect of various model carbohydrates on growth on cellobiose and Avicel. Some compounds like xylooligosaccharides caused a decline in cellulolytic activity and a reduction in the maximum solubilization rate on Avicel. However, there were no relevant model compounds that could replicate the strong inhibition by spent broth on C. thermocellum growth on cellobiose. Cocultures of C. thermocellum with hemicellulose-consuming partners—Herbinix spp. strain LL1355 and Thermoanaerobacterium thermosaccharolyticum—exhibited lower levels of unfermented hemicellulose hydrolysis products, a doubling of the maximum solubilization rate, and final solubilization increased from 67 to 93%.ConclusionsThis study documents inhibition of C. thermocellum with increasing corn fiber concentration and demonstrates inhibition of cellulase activity by xylooligosaccharides, but further work is needed to understand why growth on cellobiose was inhibited by corn fiber fermentation broth. Our results support the importance of hemicellulose-utilizing coculture partners to augment C. thermocellum in the fermentation of lignocellulosic feedstocks at high solids loading.

Effects of king grass and sugarcane top in the absence or presence of exogenous enzymes on the growth performance and rumen microbiota diversity of goats

In the present study, the feasibility of sugarcane top (ST) application in the goat's diet was evaluated. A total of 20 goats were randomly divided into four groups. The dietary treatments were set as follows: animals were fed with king grass (KG), KG with exogenous enzymes (KGE), ST, and ST with exogenous enzymes (STE). The animals were given free access to feed and water. After 15days of adaptation and 60days of the experiment, the growth performance, plasma parameters, and rumen microbiota of goats were assessed. The results showed that the KG, ST, and exogenous enzyme supplement had no significant effects on the growth performance and plasma parameters. The diet affected the rumen microbiota diversity and structure, and the alpha and beta diversity in the animals fed with ST were dramatically greater compared with the animals fed with KG. The abundances of Proteobacteria, Cyanobacteria, and Elusimicrobia were significantly decreased in the animals fed with KG or KGE, while the abundances of Firmicutes and Euryarchaeota were significantly higher in the animals fed with KG or KGE. Furthermore, the microbial communities were also different at the genus level. Moreover, the exogenous enzymes had a slight effect on rumen microbiota. Linear discriminant analysis effect size (LEfSe) analysis showed that the greatest differences were found in bacterial taxa, and these specific taxa could be used as biomarkers to distinguish rumen microbiota. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) functional profile prediction indicated that the dietary treatments of ST and KG could also alter the gene expression pattern in nutrient and energy metabolism, as well as replication and repair of genetic information pathways. Collectively, the dietary treatments of KG and ST in the absence or presence of exogenous enzymes had similar effects on the growth performance and plasma parameters of goats. Besides, the KG and ST diets could affect the rumen microbiota community and function of goats. Therefore, ST could be used as a promising alternative feed resource for ruminants without the addition of exogenous enzymes in tropical regions.

Improvement of Bioethanol Production in Consolidated Bioprocessing (CBP) via Consortium of Aspergillus niger B2484 and Trichoderma asperellum B1581

Consolidated bioprocessing (CBP) in bioethanol production involves the combination of four essential biological procedures in a single bioreactor, using a mixture of organisms with favourable cellulolytic ability without the addition of exogenous enzymes. However, the main disadvantage of this process is the complexity to optimise all factors considering both enzymes and microbial activity at the same time. Hence, this study aimed to optimise suitable culture conditions for both organisms to work efficiently. Six single factors that are considered crucial for bioethanol production were tested in one-factor-at-a-time (OFAT) analysis and analysed using Response Surface Methodology (RSM) software for Aspergillus niger B2484 and Trichoderma asperellum B1581 strains. The formulation of a new consortia setting was developed based on the average of two settings generated from RSM testing several combinations of consortia concentrations (5:1, 2:4, 3:3, 4:2, and 1:5). The combination of 5:1 Aspergillus niger B2484 and Trichoderma asperellum B1581 produced the most ethanol with 1.03 g/L, more than A. niger B2484, alone with 0.34 g/L of ethanol, indicating the potential of the combination of A. niger B2484 and T. asperellum B1581 co-culture for bioethanol production in CBP.

Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides.

Canola meal (CM), the protein-rich by-product of canola oil extraction, has shown promise as an alternative feedstuff and protein supplement in poultry diets, yet its use has been limited due to the abundance of plant cell wall fibre, specifically non-starch polysaccharides (NSP) and lignin. The addition of exogenous enzymes to promote the digestion of CM NSP in chickens has potential to increase the metabolizable energy of CM. We isolated chicken cecal bacteria from a continuous-flow mini-bioreactor system and selected for those with the ability to metabolize CM NSP. Of 100 isolates identified, Bacteroides spp. and Enterococcus spp. were the most common species with these capabilities. To identify enzymes specifically for the digestion of CM NSP, we used a combination of glycomics techniques, including enzyme-linked immunosorbent assay characterization of the plant cell wall fractions, glycosidic linkage analysis (methylation-GC-MS analysis) of CM NSP and their fractions, bacterial growth profiles using minimal media supplemented with CM NSP, and the sequencing and de novo annotation of bacterial genomes of high-efficiency CM NSP utilizing bacteria. The SACCHARIS pipeline was used to select plant cell wall active enzymes for recombinant production and characterization. This approach represents a multidisciplinary innovation platform to bioprospect endogenous CAZymes from the intestinal microbiota of herbivorous and omnivorous animals which is adaptable to a variety of applications and dietary polysaccharides.

Uncovering the biofumigant capacity of allyl isothiocyanate from several Brassicaceae crops against Fusarium pathogens in maize

Driven by environmental concerns, chemical fumigants are no longer allowed in many countries. Therefore, other strategies for reducing fungal inoculum in soils and on crop debris are being explored. In the present study, several Brassicaceae crops were screened for their potential to control Fusarium gramineaum and Fusarium poae mycelial growth in an in vitro inverted Petri dish experiment. Volatile production was measured using gas chromatography-mass spectrometry headspace analysis. A selection of cultivars from each crop species was further investigated using a pot experiment with maize. Ethiopian mustard (Brassica carinata) and brown mustard (Brassica juncea) released volatile allyl isothiocyanate (AITC) and a higher concentration of AITC was correlated with a better fungal growth reduction in the in vitro screening. Brown mustard cultivar Etamine completely inhibited growth of both Fusarium spp. Pure AITC in a solution with methanol resulted in a sigmoid dose-response curve for both Fusarium spp. tested. Fusarium poae appeared to be more tolerant to AITC than F. graminearum. A pot experiment revealed that the incorporation of brown mustard plant material could alleviate the clear negative effect of F. graminearum infection on maize growth. The present study demonstrated the correlation between the fungistatic effect of biofumigation crops on Fusarium spp. and their production of volatile AITC in vitro, without the addition of exogenous enzymes, and confirmed the biofumigation potential of brown mustard in a pot experiment with maize. These results may help farmers when selecting a green manure crop suitable for biofumigation. © 2020 Society of Chemical Industry.

Impact of dietary incorporation of Spirulina (Arthrospira platensis) and exogenous enzymes on broiler performance, carcass traits, and meat quality

This study assessed the effect of Spirulina (Arthrospira platensis), individually and in combination with exogenous enzymes, on growth performance, carcass traits, and meat quality of broiler chickens. One hundred and twenty Ross 308 male chickens were allocated into 40 battery brooders, with 3 birds per cage, and fed ad libitum a corn-based diet during the first 21 D of the trial. The experimental period lasted from day 21 to 35, during which birds were fed 4 different diets: a corn-soybean basal diet, taken as the control group, a basal diet containing 15% Spirulina (MA), a basal diet containing 15% Spirulina plus 0.005% Rovabio Excel AP (MAR), and a basal diet containing 15% Spirulina plus 0.01% lysozyme (MAL). Body weight gain (P < 0.001) and feed conversion rate (P < 0.001) were improved in control chickens, when compared with those fed with Spirulina. In addition, Spirulina increased the length of duodenum plus jejunum in relation to the other treatment (P < 0.01). Chickens on the MAL diet showed a considerable increase in digesta viscosity (P < 0.05) compared with the control group. Breast and thigh meats from chickens fed with Spirulina, with or without the addition of exogenous enzymes, had higher values of yellowness (b*) (P < 0.001), total carotenoids (P < 0.001), and saturated fatty acids (P < 0.001), whereas n-3 polyunsaturated fatty acid (P < 0.01) and α-tocopherol (P < 0.001) decreased, when compared with the control. In conclusion, the incorporation of 15% Spirulina in broiler diets, individually or combined with exogenous enzymes, reduced birds' performance through a higher digesta viscosity, which is likely associated with the gelation of microalga indigestible proteins. In addition, cell wall of Spirulina was successfully broken by the addition of lysozyme, but not by Rovabio Excel AP. Therefore, we anticipate that the combination of lysozyme with an exogenous specific peptidase could improve the digestibility of proteins from this microalga and avoid their detrimental gelation.

Effect of Zinc Source and Exogenous Enzymes Supplementation on Zinc Status in Dogs Fed High Phytate Diets.

Simple SummaryPet food is a sector in expansion, demanding novel solutions for old issues. The mineral supplementation of animal feed has been a target of research for livestock animals, but less attention has been devoted to companion animals. The present study offers both nutritionists and pet food manufacturers knowledge concerning organic and inorganic sources of zinc for dog food supplementation. Zinc proteinate and zinc sulfate were added to a high phytate dry food for healthy dogs at maintenance. Moreover, the role of exogenous enzymes to overcome the undesirable effects of phytates was also studied, with the inclusion of a solid-state fermentation product from Aspergillus niger. Data suggest that organic zinc is promising for dog food supplementation. Yet, the use of exogenous enzymes seems to require refinement to fit the needs and physiological conditions of dogs and thus add value to mineral availability and nutrient digestibility. Zinc is an essential element, a cofactor of many enzymes, and performs catalytic, structural and regulatory functions. Once in the gastrointestinal tract, zinc can interact with food constituents. Phytic acid, the major phosphorus storage in plants, limits zinc availability from animal feeds due to the formation of insoluble complexes with phytates. This study tested the effect of supplemental zinc source (zinc sulfate and a chelate zinc proteinate) and the addition of exogenous enzymes from a solid-state fermentation product of Aspergillus niger to a high phytate diet. The study was designed according to three Latin Squares 4 × 4 with a 2 × 2 factorial arrangement of treatments, with four periods, four diets, and 12 young adult Beagles. Periods lasted 5 weeks each. Diets were supplemented with 75 mg/kg of zinc sulfate (IZ) or zinc proteinate (OZ), and without or with 200 mg/kg of exogenous enzymes (IZ+, OZ+). Results showed that zinc proteinate increased the bioavailability of phosphorus, yet the zinc biomarkers remained unaffected by the zinc source, with the exception of lymphocyte subsets that benefit from zinc proteinate. The use of exogenous enzymes did not affect zinc availability nor nutrient and energy digestibility.