Impact of high Spirulina diet, extruded or supplemented with enzymes, on blood cells, systemic metabolites, and hepatic lipid and mineral profiles of broiler chickens.

The impact of 15% dietary inclusion of Spirulina (Arthrospira platensis) in broiler chickens was explored, focusing on blood cellular components, systemic metabolites and hepatic lipid and mineral composition. From days 14 to 35 of age, 120 broiler chickens were divided and allocated into four dietary treatments: a standard corn and soybean meal-based diet (control), a 15% Spirulina diet, a 15% extruded Spirulina diet, and a 15% Spirulina diet super-dosed with an enzyme blend (0.20% porcine pancreatin plus 0.01% lysozyme). The haematological analysis revealed no significant deviations (p > 0.05) in blood cell counts across treatments, suggesting that high Spirulina inclusion maintains haematological balance. The systemic metabolic assessment indicated an enhanced antioxidant capacity in birds on Spirulina diets (p < 0.001), pointing toward a potential reduction in oxidative stress. However, the study noted a detrimental impact on growth performance metrics, such as final body weight and feed conversion ratio (both p < 0.001), in the Spirulina-fed treatments, with the super-dosed enzyme blend supplementation failing to alleviate these effects but with extrusion mitigating them. Regarding hepatic composition, birds on extruded Spirulina and enzyme-supplemented diets showed a notable increase in n-3 fatty acids (EPA, DPA, DHA) (p < 0.001), leading to an improved n-6/n-3 PUFA ratio (p < 0.001). Despite this positive shift, a reduction in total hepatic lipids (p = 0.003) was observed without a significant change in cholesterol levels. Our findings underscore the need for further exploration into the optimal inclusion levels, processing methods and potential enzymatic enhancements of Spirulina in broiler diets. Ultimately, this research aims to strike a balance between promoting health benefits and maintaining optimal growth performance in poultry nutrition.

The action of endo-xylanase and endo-glucanase on cereal cell wall polysaccharides and its implications for starch digestion kinetics in an in vitro poultry model.

Endo-xylanase and endo-glucanase are supplemented to poultry diets in order to improve nutrient digestion and non-starch polysaccharide (NSP) fermentation. Here, the action of these enzymes on alcohol insoluble solids (AIS) from wheat and maize grains as well as its implications for starch digestion in milled grains were evaluated in vitro, under conditions mimicking the poultry digestive tract. For wheat AIS, GH11 endo-xylanase depolymerized soluble arabinoxylan (AX) during the gizzard phase, and proceeded to release insoluble AX under small intestine conditions. At the end of the in vitro digestion (480 min), the endo-xylanase, combined with a GH7 endo-ß-1,4-glucanase, released 30.5 % of total AX and 18.1 % of total glucan in the form of arabinoxylo- and gluco-oligosaccharides, as detected by HPAEC-PAD and MALDI-TOF-MS. For maize AIS, the combined enzyme action released 2.2 % and 7.0 % of total AX and glucan, respectively. Analogous in vitro digestion experiments of whole grains demonstrated that the enzymatic release of oligomers coincided with altered grain microstructure, as examined by SEM. In the present study, cell wall hydrolysis did not affect in vitro starch digestion kinetics for cereal grains. This study contributes to understanding the action of feed enzymes on cereal NSP under conditions mimicking the poultry digestive tract.

The fate of insoluble arabinoxylan and lignin in broilers: Influence of cereal type and dietary enzymes.

Insoluble fiber degradation by supplemented enzymes was previously shown to improve fermentation in poultry, and has been further postulated to disrupt the cereal cell wall matrix, thus improving nutrient digestion. Here, we characterized insoluble feed-derived polysaccharides and lignin in digesta from broilers fed wheat-soybean and maize-soybean diets without or with xylanase/glucanase supplementation. Enzyme supplementation in wheat-soybean diet increased the yield of water-extractable arabinoxylan (AX) in the ileum. Still, most AX (> 73 %) remained insoluble across wheat-soybean and maize-soybean diets. Analysis of so-far largely ignored lignin demonstrated that a lignin-rich fiber fraction accumulated in the gizzard, while both insoluble AX and lignin reaching the ileum appeared to be excreted unfermented. More than 20 % of water-insoluble AX was extracted by 1 M NaOH and 11-20 % was sequentially extracted by 4 M NaOH, alongside other hemicelluloses, from ileal digesta and excreta across all diets. These findings showed that enzyme-supplementation did not impact AX extractability by alkali, under the current experimental conditions. It is, therefore, suggested that the degradation of insoluble AX by dietary xylanase in vivo mainly results in arabinoxylo-oligosaccharide release, which is not accompanied by a more loose cell wall architecture.

Cereal type and combined xylanase/glucanase supplementation influence the cecal microbiota composition in broilers.

Dietary fiber-degrading enzyme supplementation in broilers aims at off-setting the anti-nutritive effect of non-starch polysaccharides and at promoting broiler health. Recently, we demonstrated that xylanase/glucanase addition in wheat-based diet improved nutrient digestibility, arabinoxylan fermentability and broiler growth. Conversely, maize arabinoxylan was found to be recalcitrant to xylanase action. These findings suggested that enzyme-mediated improvement of nutrient digestion and carbohydrate fermentation depended on the cereal type present in the diet, and may have contributed to broiler growth. Hence, we aimed at further investigating the link between dietary enzymes and carbohydrate fermentation in broilers, by studying the impact of enzyme supplementation in cereal-based diets, to the microbial communities in the ileum and ceca of broilers. For that purpose, 96 one-day-old male broilers were randomly reared in two pens and received either wheat-based or maize-based starter and grower diets. At d 20, the broilers were randomly assigned to one out of four dietary treatments. The broilers received for 8 d the wheat-based or maize-based finisher diet as such (Control treatments; WC, MC) or supplemented with a xylanase/glucanase combination (Enzyme treatments; WE, ME). At d 28, samples from the digestive tract were collected, and the ileal and cecal microbiota composition was determined by 16S ribosomal RNA gene amplicon sequencing. A similar phylogenetic (alpha) diversity was observed among the four treatments, both in the ileal and the cecal samples. Furthermore, a similar microbial composition in the ileum (beta diversity) was observed, with lactobacilli being the predominant community for all treatments. In contrast, both cereal type and enzyme supplementation were found to influence cecal communities. The type of cereal (i.e., wheat or maize) explained 47% of the total variation in microbial composition in the ceca. Further stratifying the analysis per cereal type revealed differences in microbiota composition between WC and WE, but not between MC and ME. Furthermore, the prevalence of beneficial genera, such as Faecalibacterium and Blautia, in the ceca of broilers fed wheat-based diets coincided with arabinoxylan accumulation. These findings indicated that fermentable arabinoxylan and arabinoxylo-oligosaccharides released by dietary xylanase may play an important role in bacterial metabolism.

The role of feed enzymes in maintaining poultry intestinal health.

Gut health or intestinal health is frequently discussed without any clear definition as to its meaning. It is suggested that this should be defined as intestinal integrity and functionality as both are a pre-requisite for the health of the intestine itself and the host. The health of the intestine is dependent upon a successful evolution of the absorptive capacity of the intestine, which in turn is influenced by the co-evolution of the intestinal immune systems and the microbiota. Nutrient supply plays a significant role in this process and from the perspective of the microbiota this changes with age as the intestines and upper gastrointestinal tract (GIT) microbiota become more effective in nutrient removal. Feed enzymes play a significant role in this process. Phytases can improve digestion of minerals, amino acids and energy and as a result reduce the availability of nutrients in the lower intestines for the microbiota. Protease can have a similar effect with amino acid supply. Non-starch polysaccharidases (NSPases) have a unique role in that they not only improve diet digestibility from the hosts perspective, thus limiting nutrient supply to the microbiota, but they also release soluble fragments of fibre from the insoluble matrix and/or depolymerize high molecular weight viscous fibre fractions in to smaller, more fermentable carbohydrate fractions. This results in a more favourable balance between fermentable carbohydrate to protein supply, a ratio which is deemed critical to maintaining good intestinal health. The dynamic nature of this complex evolution needs greater consideration if antibiotic free production is to succeed. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Amino acids and energy digestibility in extruded or roasted full fat soybean fed to broiler chickens without or with multienzyme supplement containing protease, phytase, and fiber degrading enzymes.

Effects of processing and multienzyme supplement (MES) on standardized ileal digestibility (SID) of amino acids, apparent retention (AR) of components and metabolizable energy (AME) content in full fat soybean seeds (FFSB) were investigated in broiler chickens. The FFSB were either extruded (EFFSB) or roasted (RFFSB). A nitrogen free diet (NFD) was formulated for SID of AA calculation. The FFSB diets contained 20% crude protein with the ratio of corn starch: sucrose: soy oil (sole sources of energy in NFD) kept constant for calculation of AME. The FFSB diets were fed without or with MES containing phytase, protease, and fiber degrading enzymes. All diets had TiO2 indigestible marker. A total of 400-dayu-old Ross 708 male chicks were fed a commercial diet to d 13. On d 14, birds were weighed individually and allocated to cages (10 birds/cage, n = 8). Birds had free access to feed and water. Excreta samples were collected on d 18 to 20, and all birds were necropsied on d 21 for terminal ileal digesta samples. There was no (P > 0.05) interaction between processing and MES on SID of AA. Birds fed EFFSB had higher (P ≤ 0.048) SID of Arg, Ile, Lys, and Met than birds fed RFFSB. Birds fed MES had higher (85.5 vs. 80.8%; P = 0.050) SID of Lys than birds fed non-MES diet. There was interaction (P ≤ 0.036) between processing and MES on AR of Ca and P; MES improved retention but largely in EFFSB. There was an interaction (P = 0.016) between processing and MES on energy utilization such that MES improved AR of GE, AME, and AMEn in RFFSB only. In general, birds fed EFFSB exhibited higher (P < 0.01) energy utilization than birds fed RFFSB. In conclusion, lower Lys and energy utilization in RFFSB relative to EFFSB reflected the impact of the processing regimen. Supplemental enzyme improvement on Lys and minerals digestibility in FFSB and energy utilization in RFFSB suggested value in heat processed feedstuffs.

Growth performance, organ attributes, nutrient and caloric utilization in broiler chickens differing in growth rates when fed a corn-soybean meal diet with multienzyme supplement containing phytase, protease and fiber degrading enzymes.

Growth performance, organ weight, ceca digesta short chain fatty acids (SCFA), jejunal histomorphometry, tibia ash, apparent retention (AR) of components and caloric efficiency were investigated in broiler chicken strains differing in growth rate fed diets with multienzyme supplement (MES). The strains differed in estimated time to reach 2.1 kg BW: 37, 43, 47, and 50 d and were designated C, F, J, and N, respectively. A corn-soybean meal diet was formulated for 2-phase program (starter and grower) and fed without or with MES containing phytase, protease and fiber-degrading enzymes. A total of 640-day-old chicks (42.3 + 0.01 g/bird) were housed in cages (5 cockerels and 5 pullets/cage) and allocated to give 8 replicates/ strain and diet combination. Equal amount of feed was fed based on observed ad-libitum intake of C strain in the starter (d 0-14) and grower (d 15-28). Body weight was monitored, grab excreta samples taken and at completion of allocated feed one bird per cage necropsied for samples. With exception of P, apparent metabolizable energy corrected for nitrogen (AMEn) and ceca digesta acetic acid, there was no (P > 0.05) interaction between strain and MES on examined responses. Strains differed (P < 0.01) on growth, FCR, gizzard weight, tibia ash, breast weight, ceca digesta concentration of lactic, propionic, and isobutyric acid and caloric efficiency. The final body weight (BW) was 1,344, 1,134, 959, and 916 g/bird for C, F, J, and N, respectively. Corresponding caloric efficiency was 4,930, 5,807, 6,680 and 7,199 kcal/kg BW gain, respectively. Birds fed MES had higher BW gain (P < 0.05) in grower phase, larger gizzard, higher AR of CP, crude fat, neutral detergent fiber, and Ca than non-MES birds. In conclusion, growth rate influenced organ attributes, nutrient, and caloric utilization. Enzyme supplementation improved growth in grower phase and nutrient utilization independent of strain, suggesting that effects of feed enzymes are not influenced by inherent growth rate.

Effects of feeding corn distillers dried grains with solubles diets without or with supplemental enzymes on growth performance of pigs: a meta-analysis.

A meta-analysis was conducted to determine the effects of the dietary energy system (net energy or metabolizable energy), oil content of corn distillers dried grains with solubles (cDDGS), diet inclusion levels, and pig age on growth performance of pigs fed cDDGS-based diets. Mean differences of average daily gain (ADG), average daily feed intake (ADFI), and gain:feed (G:F) were calculated and expressed as a percentage change relative to feeding corn-soybean meal (SBM)- and cDDGS-based diets to nursery [body weight (BW) < 25 kg] and growing-finishing (BW > 25 kg) pigs, and to compare the effects of supplementing various types of exogenous enzymes without or with phytase on growth performance. A total of 27 studies with 106 growth performance observations were included in the cDDGS dataset, and 34 studies with 84 observations for enzyme responses in cDDGS diets were used in the enzyme dataset. Approximately, 64.7% of the observations showed no change, and 26.7% of observations showed a reduction in ADG, ADFI, and G:F when feeding cDDGS-based diets to the nursery and growing-finishing pigs compared with feeding corn-SBM-based diets. Furthermore, feeding cDDGS diets resulted in decreased (P < 0.01) mean difference of ADG (-4.27%) and G:F (-1.99%) for nursery pigs, and decreased (P < 0.01) mean difference of ADG (-1.68%) and G:F (-1.06%) for growing-finishing pigs. Every percentage unit increase in the inclusion level of cDDGS in growing-finishing pig diet was associated with a decrease (P < 0.01) in ADG (-0.10%) and ADFI (-0.09%). Feeding high oil (≥10% ether extract) cDDGS-based diets to pigs resulted in a 2.96% reduction in ADFI whereas feeding reduced-oil (<10% ether extract) cDDGS-based diets reduced G:F by 1.56% compared with pigs fed corn-SBM-based diets. Supplementation of exogenous enzymes improved (P < 0.05) the mean difference of ADG and G:F by 1.94% and 2.65%, respectively, in corn-SBM-based diets, and by 2.67% and 1.87%, respectively, in cDDGS diets. Supplementation of exogenous protease, enzyme cocktail, or xylanase improved (P < 0.05) ADG by 7.29%, 2.64%, and 2.48% in pigs fed corn-SBM-based diets, respectively. There were no differences between the dietary addition of single enzymes and enzyme combinations for any growth performance parameters in corn-SBM- or cDDGS-based diets. In conclusion, feeding cDDGS-based diets slightly reduces the growth performance of nursery and growing-finishing pigs, but supplementation of xylanase or enzyme cocktail can improve G:F of pigs fed cDDGS-based diets.

Effects of Chlorella vulgaris as a Feed Ingredient on the Quality and Nutritional Value of Weaned Piglets' Meat.

Chlorella vulgaris (CH) is usually considered a feed supplement in pig nutrition, and its use as an ingredient is poorly studied. Among many interesting characteristics, this microalga has high protein levels and can be a putative alternative for soybean meal. Our aim was to study the effect of a 5% CH incorporation in the diet, individually or combined with two carbohydrases, on meat quality traits and nutritional value. Forty-four post-weaned male piglets individually housed, with an initial live weight of 11.2 ± 0.46 kg, were randomly distributed into four experimental groups: control (n = 11, without CH) and three groups fed with 5% CH incorporation, plain (n = 10), with 0.005% Rovabio® Excel AP (n = 10), and with 0.01% of a pre-selected four-CAZyme mixture (n = 11). After two weeks of trial, piglets were slaughtered and longissimus lumborum collected. CH had no effect on piglets' growth performance. In turn, incorporation of CH improved the nutritional value of meat by increasing total carotenoids and n-3 PUFA content, thus contributing to a more positive n-6/n-3 fatty acid ratio. The supplementation with Rovabio® benefited tenderness and increased overall acceptability of pork. Our results show beyond doubt the viability of the utilization of this microalga as a feed ingredient for swine production.

Impact of Xylanase and Glucanase on Oligosaccharide Formation, Carbohydrate Fermentation Patterns, and Nutrient Utilization in the Gastrointestinal Tract of Broilers.

This study aimed at determining how the degradation of cereal non-starch polysaccharides (NSP) by dietary enzymes during feed digestion can influence nutrient digestibility and NSP fermentability in broilers. Ninety-six one-day-old male broilers were assigned to 4 different treatments: control and enzyme-supplemented wheat-based (WC, WE) or maize-based (MC, ME) treatments. Enzyme supplementation with endo-xylanase and endo-glucanase occurred from day 20 onwards. On day 28, digesta samples were collected. Nutrient digestibility, NSP recovery, oligosaccharide profile, and short-chain fatty acids (SCFA) content were determined. Enzyme supplementation in WE resulted in a higher starch (3%; p = 0.004) and protein (5%; p = 0.002) digestion in the ileum compared to WC. Xylanase activity in WE led to in situ formations of arabinoxylan-oligosaccharides consisting of 5 to 26 pentose units in the ileum. This coincided with decreased arabinose (p = 0.059) and xylose (p = 0.036) amounts in the ceca and higher acetate (p = 0.014) and butyrate (p = 0.044) formation in WE compared to WC. Conversely, complete total tract recovery of arabinoxylan in MC and ME suggested poor maize NSP fermentability. Overall, enzyme action improved nutrient digestibility and arabinoxylan fermentability in the wheat-based diet. The lower response of the maize-based diet to enzyme treatment may be related to the recalcitrance of maize arabinoxylan as well as to the high nutritive value of maize.

Improving the digestibility of cereal fractions of wheat, maize, and rice by a carbohydrase complex rich in xylanases and arabinofuranosidases: an in vitro digestion study.

BACKGROUND: Cereal co-products rich in dietary fibres are increasingly used in animal feed. The high fibre content decreases the digestibility and reduces the nutrient and energy availability, resulting in lower nutritive value. Therefore, this study investigated the ability of two carbohydrase complexes to solubilize cell-wall polysaccharides, in particular arabinoxylan (AX), from different cereal fractions of wheat, maize, and rice using an in vitro digestion model of the pig gastric and small intestinal digestive system. The first complex (NSPase 1) was rich in cell-wall-degrading enzymes, whereas the second complex (NSPase 2) was additionally enriched with xylanases and arabinofuranosidases. The extent of solubilization of insoluble cell-wall polysaccharides after in vitro digestion was evaluated with gas-liquid chromatography and an enzymatic fingerprint of the AX oligosaccharides was obtained with high-performance anion-exchange chromatography with pulsed amperometric detection. RESULTS: The addition of carbohydrase increased the digestibility of dry matter and solubilized AX in particular, with the greatest effect in wheat fractions and less effect in maize and rice fractions. The solubilization of AX (expressed as xylose release) ranged from 6% to 41%, and there was an increased effect when enriching with xylanases and arabinofuranosidases in wheat aleurone and bran of 19% and 14% respectively. The enzymatic fingerprint of AX oligosaccharides revealed several non-final hydrolysis products of the enzymes applied, indicating that the hydrolysis of AX was not completed during in vitro digestion. CONCLUSION: These results indicate that the addition of a carbohydrase complex can introduce structural alterations under in vitro digestion conditions, and that enrichment with additional xylanases and arabinofuranosidases can boost this effect in wheat, maize, and rice. © 2020 Society of Chemical Industry.

Are antimicrobial resistance genes key targets to detect genetically modified microorganisms in fermentation products?

As genetically modified microorganisms (GMM), commonly used by the food and feed industry to produce additives, enzymes and flavourings, are frequently harbouring antimicrobial resistance (AMR) genes as selection markers, health and environmental concerns were consequently raised. For this reason, the interest of the competent authorities to control such microbial fermentation products has strongly increased, especially since several recent accidental contaminations of unauthorized GMM, or associated recombinant DNA, in bacterial fermentation products intended for the European food and feed chain. However, no global screening strategy is currently available in enforcement laboratories to assess the presence of GMM harbouring AMR genes and/or the presence of full-length AMR genes. Moreover, the confidentiality of the related GMM dossiers strongly hampers the development of methods to perform such control. To overcome this issue, an analysis of related publicly available patents was performed in this study to identify all reported AMR genes. On this basis, the aminoglycoside adenyltransferase (aadD) gene, conferring a resistance to both kanamycin and neomycin, was identified as a key target to cover a large spectrum of GM bacteria. A real-time PCR method to screen for its potential presence as well as a nested-PCR method associated with a sequencing analysis to assess its full-length were developed to target this aadD gene. The performance of these new methods were successfully evaluated in terms of specificity, sensitivity and applicability, allowing their easy implementation in enforcement laboratories. Moreover, the integration of these newly developed methods to our very recently proposed strategy, initially targeting GMM carrying a chloramphenicol resistance gene, allows to drastically increase the detection spectrum of GM bacteria producing fermentation food and feed products. The data generated by the proposed strategy represents therefore a crucial support for the competent authorities, especially to evaluate potential risks for the food and feed safety.

Effects of feeding broiler breeder hens a coextruded full-fat flaxseed and pulses mixture without or with multienzyme supplement.

The effects of coextruded full-fat flaxseed and pulses (FFF; 1:1 wt/wt) mixture on n-3 polyunsaturated fatty acids (PUFA) enrichment in egg yolk, hepatic attributes, apparent retention (AR) of components, and ceca metabolites were evaluated in broiler breeder hens. The diets were as follows: 1) corn-soybean control, 2) control diet plus 18% FFF (FFF-), and 3) FFF plus enzyme supplement (FFF+) containing galactanase, protease, mannanase, glucanase, xylanase, amylase, and cellulase activities. Twenty-six-week-old Cobb 500 broiler breeder hens were allocated to 30 identical cages (2 hens/cage) and given 1-week adaptation period. The 3 diets were assigned to 10 replicate cages based on postadaptation BW and fed based on breeder curve for 30 D. Excreta samples were collected from day 24 to 27 for determination of AR of components, and eggs were collected from day 28 to 30 for yolk polyunsaturated fatty acids analyses. On day 30, birds were weighed, killed via cervical dislocation, liver weighed, and stored for fat analyses. Ceca digesta samples were taken for concentration of short-chain fatty acids. Liver and yolk weights as well as total yolk FA were not influenced by diets (P > 0.05). Control birds had lower yolk concentration of α-linolenic acid than birds fed either FFF- or FFF+ (P < 0.01) corresponding to 7.5, 36.8, and 37.3 mg/g for the control, FFF-, and FFF+, respectively. Control birds also exhibited lower yolk concentration of docosahexaenoic acid (P < 0.01). Control birds had higher hepatic concentration of crude fat and apparent retention of dry matter and crude protein compared with either the FFF- or FFF+ birds (P < 0.05). Birds fed FFF- diet had lower ceca digesta concentration of lactic acid than control and FFF+ (P < 0.05) birds. Results showed broiler breeder hens enriched egg yolk with n-3 polyunsaturated fatty acids without effects on the liver while the supplemental enzyme did not improve the utilization of FFF.

Dietary β-mannanase decreases cloacal temperature of broiler chickens under hot conditions without affecting growth performance / El consumo de β-mananasa disminuye la temperatura cloacal de pollos de engorde bajo condiciones de calor sin afectar su rendimiento de crecimiento / O consumo de β-mananase diminui a temperatura da cloaca do frango de corte em condições de calor sem afetar seu desempenho do crescimento

Abstract Background: High amounts of nonstarch polysaccharides in the diet may increase the amounts of fermentative materials in the hindgut, leading to an increase in fermentative heat production. Dietary β-mannanase is reported to decrease antinutritional effects of β-mannans, such as the potential increase of body heat; however, its efficacy on broiler chickens raised under hot climatic conditions has not been investigated. Objective: To investigate the effects of dietary β-mannanase on growth performance, cloacal temperature, relative lymphoid organ weight, and blood characteristics of broiler chickens raised under hot climatic conditions. Methods: A total of 1,701 1-day-old Ross 308 broiler chickens were randomly allotted to one of three dietary treatments with nine replicates. A basal diet was prepared and added with β-mannanase at 0.05 or 0.10% inclusion levels. The experiment was conducted for 30 days. Average room temperature was 28.8 ± 1.74 ˚C and average relative humidity (RH) was 76.1 ± 11.49% during the experiment. Results: Growth performance of broiler chickens raised under hot climatic conditions was not affected by β-mannanase inclusion. Cloacal temperature decreased at the end of experiment (linear, p<0.05) with increasing inclusion levels of dietary β-mannanase. Increasing inclusion levels of β-mannanase tended to increase (linear, p=0.076) the relative weight of thymus, but had no effects on the relative weight of spleen and bursa of Fabricius. Blood characteristics were not influenced by dietary β-mannanase. Conclusion: Increasing inclusion levels of β-mannanase decrease cloacal temperature; however, it does not directly influence growth performance nor alleviates the heat stress of broiler chickens raised under hot climatic conditions.

Resumen Antecedentes: Altas cantidades de carbohidratos no almidonosos en la dieta pueden aumentar la cantidad de materiales fermentativos en el intestino posterior, aumentando la producción de calor fermentativo. La β-mananasa dietaria disminuye los efectos antinutricionales de los β-mananos, tales como el potencial incremento de la producción calorica; Sin embargo, no se ha investigado su eficacia en pollos de engorde criados bajo condiciones de calor ambiental. Objetivo: Investigar los efectos de la β-mananasa en la dieta sobre el crecimiento, la temperatura cloacal, el peso relativo de los órganos linfoides y las características sanguíneas de pollos de engorde criados bajo condiciones climáticas calientes. Métodos: Un total de 1.701 pollos de engorde de 1 día de edad (Ross 308) fueron asignados al azar a uno de tres tratamientos dietarios con nueve repeticiones. A una dieta basal se le adicionó β-mananasa en niveles de inclusión de 0,05 o 0,10%. El experimento duró 30 días. La temperatura ambiente durante el experimento fue de 28,8 ± 1,74 ˚C y la humedad relativa de 76,1 ± 11,49%. Resultados: La inclusión de β-mananasa no afecto el rendimiento de los pollos. La temperatura cloacal medida al final del experimento disminuyó (lineal, p<0,05) con niveles de inclusión dietarios crecientes de β-mananasa. Niveles incrementales de β-mananasa tendieron a aumentar (lineal, p=0,076) el peso relativo del timo, pero no hubo efecto sobre el peso del bazo o la bursa de Fabricio. La β-mananasa no influenció las características de la sangre. Conclusión: Niveles incrementales de β-mananasa disminuyen la temperatura cloacal, aunque afectan el crecimiento ni alivian el estrés térmico del pollo de engorde criado bajo condiciones climáticas calientes.

Resumo Antecedentes: Altas quantidades de polissacáridos não amiláceos na dieta podem aumentar as quantidades de materiais fermentativos no intestino posterior, levando a um aumento na produção de calor fermentativo. A β-mananase dietética é relatada para diminuir os efeitos antinutricionais da β-manana, tal como o possível aumento da produção de calor; Entretanto, não tem sido investigada a eficácia de la em frangos de corte criados em condições climáticas quentes. Objetivo: Investigar os efeitos da β-mananase na dieta sobre o desempenho do crescimento, a temperatura cloacal, o peso relativo dos órgãos linfóides e as características de sanguíneas de frangos de corte criados em condições climáticas quentes. Métodos: Um total de 1.701 frangos de corte de um dia de idade (Ross 308) foram distribuídos aleatoriamente em um dos três tratamentos dietéticos com nove repetições. A dieta basal foi preparada e a β-mananase foi adicionada à dieta basal com níveis de inclusão de 0,05 ou 0,10%. O experimento foi conduzido durante 30 dias. A temperatura ambiente média foi de 28,8 temperatura ahumidade relativa (HR) média foi de 76,1 ± 11,49% durante o experimento. Resultados: O desempenho de crescimento dos frangos de corte criados em condições climáticas quentes não foi afetado pelos níveis de inclusão de β-mananase. A temperatura cloacal medida no final do experimento foi diminuída (linear, p<0,05) com níveis crescentes de inclusão de β-mananase na dieta. O aumento dos níveis de inclusão da β-mananase tendeu a aumentar (linear, p=0,076) o peso relativo do timo, mas não teve efeitos sobre o peso relativo do baço e da bursa de Fabricius. As características do sangue não foram influenciadas pela β-mananase dietética. Conclusão: Os níveis crescentes de inclusão de β-mananase diminuem a temperatura cloacal; entretanto, isso não influencia diretamente o desempenho do crescimento e alivia o estresse térmico dos frangos de corte criados em condições climáticas quentes.

Analysis of the Rumen Microbiota of Beef Calves Supplemented During the Suckling Phase.

A study was conducted to examine the effects of supplementing beef calves during their suckling phase (popularly known as creep feeding) with supplements that contained or did not contain the enzyme xylanase. Forty-two cow-calf pairs were divided into three groups and assigned to one of three treatments for a period of 105 days, as follows: (1) No supplemental feed for calves (control; CON); (2) Corn and soybean meal-based supplement feed for calves (positive control; PCON); and (3) Same feed regimen as PCON with xylanase added to the supplement (enzyme; ENZ). After 105 days, out of the 42 calves participating in the study, 25 male calves were randomly selected (8 from CON, 9 from PCON, and 8 from ENZ) and samples of their forestomach were collected by esophageal tubing. Immediately after this procedure, all calves were weaned, commingled, and placed in a common post-weaning diet for 4 weeks. At the end of this period, ruminal fluid was once again collected from the same 25 calves. All samples were subjected to DNA extraction and 16S rRNA gene sequencing. At weaning, most of the alpha diversity indexes were greater in CON; however, no differences (P ≥ 0.23) in alpha diversity were observed in samples collected 4 weeks after weaning. Regardless of treatment, 2 phyla - Bacteroidetes and Firmicutes - comprised approximately 80% of the total bacterial abundance of samples collected on both days. At the genus level, an effect of diet (P = 0.02) was observed for Prevotella in the samples collected at weaning; however, no differences were detected in the samples collected 4 weeks after weaning. Calf average daily gain (ADG) during the 105-day creep feeding trial tended (P = 0.09) to be greater in the groups that received supplementation, with the greatest numerical value observed in ENZ. Moreover, there was a positive correlation (ρ = 0.43; P = 0.03) between ADG and abundance of Prevotella, indicating the importance of this bacterial group for ruminants. In summary, most of the significant differences found in this study were detected at weaning, and the majority of them disappeared 4 weeks after the calves were weaned and commingled.

Role of Feed Processing on Gut Health and Function in Pigs and Poultry: Conundrum of Optimal Particle Size and Hydrothermal Regimens.

The aim is to give an overview of available literature data on the role of feed processing on gut health and function with specific focus on particle size and hydrothermal processing. In addition, influence of feed processing on efficacy of exogenous feed enzymes will be discussed. The current feed processing technologies are such that ingredient choices and diet form are refined to improve feed intake and nutrient utilization efficiency. Finer feed particle size enables optimal nutrient utilization and enhances animal performance due to increased surface area allowing better contact with digestive enzymes. Moreover, adequate diminution of feed ingredients is beneficial to feed manufacturing processes such as mixing and hydrothermal treatments including pelleting, extrusion, and expansion. However, emerging trends in consumer and regulatory demands for restriction or cessation of animal production practices such as use of antimicrobial growth promoters are challenging current approaches to feed processing. There is limit as to the fineness of the particle size, as very fine particles negatively affect gut health due to higher incidences of stomach ulceration in pigs and gizzard dysfunction in poultry. Coarse particle size increases stomach and hindgut acidification which may be beneficial in controlling proliferation of enteric pathogens such as salmonella and E. coli. Optimal particle size could be designed in the grinding process using roller or hammer mill. However, since most commercial pigs and poultry diets are subjected to hydrothermal processes, additional reduction of feed particle size is inevitable. The need to achieve high physical quality and to reduce potential levels of feed-borne pathogens such as Salmonella has led to the application of relatively high conditioning temperatures during conventional hydrothermal processes, a practice that does not favor high nutrient utilization and stability of heat sensitive feed additives such as feed enzymes. Therefore, with evolving pig and poultry production practices, the regimens for feed processing will no longer be appreciated only in terms of optimizing nutrients utilization, but also in terms of impact on feed hygienic status, efficacy of feed additives, animal health, and food safety.

Utility of Feed Enzymes and Yeast Derivatives in Ameliorating Deleterious Effects of Coccidiosis on Intestinal Health and Function in Broiler Chickens.

Coccidiosis induced necrotic lesions impair digestive capacity and barrier function in concurrence with increased risks for secondary bacterial infections. The industry has been successful in controlling coccidiosis with anticoccidials and vaccination. However, concerns over Eimeria species resistant to anticoccidials, gaps in vaccination and restriction on antibiotics is stimulating research and application of alternative and/or complimentary strategies for coccidiosis control. The aim of this paper is to appraise literature on the utility of feed enzymes and yeast derivatives in modulating coccidiosis. Feed enzymes can complement endogenous enzymes (protease, amylase, and lipase) that may become insufficient in coccidiosis afflicted birds. Coccidiosis in the upper small intestine creates conditions that enhances efficacy of phytase and there are reports indicating supplemental phytase can mitigate the negative impact of coccidiosis on bone quality. Increase in intestinal short chain fatty acids due supplemental fiber degrading enzymes has been linked with reduced survivability of Eimeria. There is evidence whole yeast (live or dead) and derivatives can modulate coccidiosis. Immunomudulation properties of the yeast derivatives have been shown to enhance cellular and humoral immunity in Eimeria challenge models which is critical for effectiveness of coccidial vaccination. Moreover, yeast nucleotides have been shown to be beneficial in stimulating healing of intestinal mucosal surface. Other novel work has shown that certain yeast cells can produce derivatives with anticoccidial compounds effective in attenuating oocysts shedding. Yeast cell surface has also been shown to be an effective oral Eimeria vaccine delivery vehicle. Overall, while further refinement research is warranted to address inconsistencies in responses and commercial application, there is evidence feed enzymes and yeast derivatives could complement strategies for maintaining intestinal function to bolster growth performance in broilers compromised with coccidiosis. However, broilers receive diets containing several feed additives with distinct mode of actions and yet there is dearth of empirical data on the expected responses.Future evaluations should consider combinations of additives to document animal responses and potential synergies.

The effect of enzymes on release of trace elements in feedstuffs based on in vitro digestion model for monogastric livestock.

BACKGROUND: This experiment was conducted to study the effect of different feed enzymes (phytase, xylanase, ß-glucanase) on release rate of trace elements (Fe, Cu, Mn and Zn) in 6 commonly used feedstuffs (corn, wheat, barley, soybean meal, wheat bran, wheat middlings) by using an in vitro model, simulating the digestive processes in stomach for 2 h and then in small intestine for 6 h at 39 °C. RESULTS: Phytase raised (P < 0.05) the release rate of Cu and Zn in corn, Cu, Zn and Mn in wheat, Cu in barley, Cu, Zn and Mn in soybean meal, Zn, Fe in wheat bran and Zn, Fe, Mn in wheat middlings. The release rate of various trace elements in feedstuffs was increased after xylanase addition. Compared with the control group, the release rate of soluble Cu in corn, wheat, barley and soybean meal, soluble Zn in corn, wheat and wheat middlings and soluble of Mn in corn, wheat, barley and wheat bran increased (P < 0.05) after xylanase treatment. After the treatment of ß-glucanase, the release rate of soluble Cu in corn, wheat and wheat bran, soluble Fe in barley, soybean meal and wheat bran and soluble Mn in corn and wheat bran all increased (P < 0.05) compared with the control group. In each feedstuff, after corresponding enzyme treatment, the contents of phytic acid, xylan and ß-glucan were significantly lower than those of the control group (P < 0.05). CONCLUSIONS: Results showed that bound trace elements in feedstuffs can be released by feed enzymes. It may be necessary to take the trace elements in feedstuffs into account in the actual feed preparation including feed enzymes.

Effects of pea extrusion and enzyme and probiotic supplementation on performance, microbiota activity and biofilm formation in the broiler gastrointestinal tract.

1. The effects of supplementation of broiler chicken diets with pea meal, carbohydrase enzymes and a probiotic were investigated for potential performance improvement. 2. Raw or extruded pea meal (cv Model, grown in Poland) was included in a wheat-soybean meal-based diet at 250 g/kg. The diets were unsupplemented (control) or supplemented with either carbohydrase enzymes (200 U/kg xylanase and 10 U/kg ß-glucanase in feed) or a probiotic (Bacillus subtilis), or both. The diets were fed to Ross 308 broilers aged 9-28 days. 3. After two additional days, chick gastrointestinal tracts were excised and analysed for the presence of Bacillus subtilis biofilm; and the ileal and caecal digesta were analysed for bacterial enzyme activities and to determine the concentration of short-chain fatty acids (SCFAs). 4. Feeding the pea-based diet supplemented with the probiotic compromised feed utilisation, due to higher feed intake. The addition of enzymes to the raw, but not the extruded, pea containing diet partially ameliorated this effect (pea form × additives; P < 0.002). 5. In the ileal digesta, interactions between the dietary treatments were observed for the activities of all bacterial glycolytic enzymes and for SCFA concentrations. ß-glucosidase, α-galactosidase and ß-glucuronidase were highest in birds fed the diet containing extruded pea supplemented with the probiotic and enzymes (pea form x additives; P = 0.018 to P < 0.006). In the caecal digesta, interactions were observed for bacterial enzyme activities, but not for total SCFA concentration. Biofilm formation in the caecum indicated that the probiotic strain was metabolically active in the broiler gut. 6. In conclusion, supplementation of diets containing raw or extruded pea meal with enzymes and a Bacillus subtilis spore-based probiotic modulated microbiota activity but had no clear effects on broiler performance. Probiotic administration did not cause excessive fermentation in the ileum and caecum but enhanced Bacillus subtilis spp. biofilm formation in the caecum, which may be indicative of a beneficial effect on gut health.

The evolution and application of enzymes in the animal feed industry: the role of data interpretation.

1. Enzymes have been used commercially for nearly 40 years and save significant costs through sparing of expensive nutrients but the mechanism by which this is achieved is still debated. 2. The research focused on non-starch polysaccharidase (NSPase) enzymes is used as an example of where greater progress could have been made if the details of the work had been described more fully and the analysis of the data generated had been broader in scope and more critical. 3. Lack of standardisation of the details presented in the materials and methods has been identified as a significant barrier to meaningful retrospective analysis and thus limits advances in the understanding of the mode of action of these enzymes. 4. The identity of the enzyme employed and its activity is often lacking, and more importantly the purity is rarely disclosed. Contaminant activities which are neither listed nor assayed could play a significant role in the responses observed. 5. The dose optimum of most enzymes is often considerably higher than that employed in most studies. Thus studies claiming synergy between two 'activities' should ensure that the response is not related to each enzyme simply augmenting the dose of just one activity in the finished feed. This is a common problem, and coupled with the lack of factorial experiments to justify the presence of each enzyme in a multi-enzyme product, it is not surprising that there is still debate as to whether single or multi-enzymes are best suited poultry rations. 6. The three proposed mechanisms for NSPases (viscosity, cell wall and prebiotic) are discussed, and along with their strengths and weaknesses it is suggested that a re-evaluation of each is needed. Viscosity may have to be re-evaluated as being a function not only of the cereal being fed, but of the age of the animal as well. The cell wall theory as described is poorly modelled in vitro and hence the validity of these data is questioned. The prebiotic theory may need significant modification as it appears that the quantities of oligomers produced are insufficient to generate the additional volatile fatty acids (VFA)'s reported. It is likely that all three mechanisms play a role in the responses observed, but the prebiotic mechanism probably plays by far the most important part in low viscosity diets. 7. Future research would be improved if it considered all potential mechanisms when designing a trial. Significant failings are apparent as a result of adherence to tenets in explanation of the results. Most importantly, it should be emphasised that a hypothesis is there to be tested, not defended.