Dietary fibre effects and the interplay with exogenous carbohydrases in poultry nutrition.

A comprehensive understanding of the role of dietary fibre in non-ruminant animal production is elusive. Equivocal and conflated definitions of fibre coupled with significant analytical complexity, interact with poorly defined host and microbiome relationships. Dietary fibre is known to influence gut development, feed intake and passage rate, nutrient absorption, microbiome taxonomy and function, gut pH, endogenous nutrient loss, environmental sustainability, animal welfare and more. Whilst significant gaps persist in our understanding of fibre in non-ruminant animal production, there is substantial interest in optimizing the fibre fraction of feed to induce high value phenotypes such as improved welfare, live performance and to reduce the environmental footprint of animal production systems. In order to achieve these aspirational goals, it is important to tackle dietary fibre with the same level of scrutiny as is currently done for other critical nutrient classes such as protein, minerals and vitamins. The chemical, mechanical and nutritional role of fibre must be explored at the level of monomeric sugars, oligosaccharides and polysaccharides of varying molecular weight and decoration, and this must be in parallel to standardisation of analytical tools and definitions for speciation. To further complicate subject, exogenous carbohydrases recognise dietary fibre as a focal substrate and have varying capacity to generate lower molecular weight carbohydrates that interact differentially with the host and the enteric microbiome. This short review article will explore the interactive space between dietary fibre and exogenous carbohydrases and will include their nutritional and health effects with emphasis on functional development of the gut, microbiome modulation and host metabolism.

The effects of concurrent increases in supplementation of calcium and phytase on growth performance, balance of Ca and P, and bone mineralization in nursery pigs.

The objective of this study was to investigate the effects of concomitantly increasing supplementation of Ca and phytase on growth performance, balance of Ca and P, and bone mineralization in nursery pigs. There were eight experimental diets. The positive control (PC) one and two were formulated to contain 0.64% and 0.85% total Ca, respectively, whereas the dietary concentrations of other nutrients were identical and adequate. The negative control (NC) was deficient in total Ca (0.48%) and total P (0.41%). Five combinations of incremental levels of Ca and phytase (0.48% and 1,750 phytase units [FYT]/kg, 0.52% and 2,000 FYT/kg, 0.55% and 2,250 FYT/kg, 0.59% and 2,600 FYT/kg, and 0.63% and 3,000 FYT/kg) were added to the NC to establish the remaining five experimental diets. Each diet was fed to six pens of six pigs (three barrows and three gilts per pen). All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen during the trial. In the end, one pig per pen was euthanized to collect the right tibia and urine in bladder. The results showed that the pigs of NC gained less weight, consumed less feed, and utilized feed less efficiently than their counterparts fed the PC and the treatments with phytase (P < 0.01). With increasing supplementation of Ca and phytase, there was a tendency for gain:feed to decrease (P < 0.10). There was a significant reduction in bone dry weight; and in percentages, as well as weights of bone ash, Ca, and P; in pigs of NC compared with pigs of PC1, PC2, or phytase treatments. In comparison to PC2, PC1 and phytase treatments resulted in a higher percentage of bone P and greater weights of bone ash, Ca, and P (P < 0.05). There was no significant effect of concurrent supplementation of Ca and phytase on bone mineralization. The NC had significantly lower apparent total tract digestibility (ATTD) of Ca and P, lower concentrations of digestible Ca and P, but a higher ATTD Ca/ATTD P ratio than PC1, PC2, or the phytase treatments. The averages of ATTD of Ca and P in treatments with phytase were significantly higher than PC1 or PC2 (P < 0.01). With increasing addition of Ca and phytase, the ATTD of P, digestible Ca and P, and the ATTD Ca/ATTD P ratio increased linearly (P < 0.05), which contrasted with a linear reduction in ATTD of Ca (P < 0.05). Meanwhile, there was a linear (P < 0.01) increase in the concentration of urinary Ca. In conclusion, increasing the dietary supplementation of phytase in conjunction with the increasing dietary Ca level increased the dietary ATTD Ca/ATTD P ratio without damaging the absorption of P in the current study. The higher ATTD Ca/ATTD P ratio did not improve the bone mineralization markedly and thus the extra Ca was voided through urine.

The effects of increasing dietary total Ca/total P ratios on growth performance, Ca and P balance, and bone mineralization in nursery pigs fed diets supplemented with phytase.

The objective of this study was to investigate the effects of increasing dietary total Ca/total P ratios on growth performance, digestibility of Ca and P, bone mineralization, and concentrations of Ca and P in urine and plasma in nursery pigs. There were six diets in a randomized complete block design, including one positive control and five diets corresponding to five total Ca/total P ratios: 0.55, 0.73, 0.90, 1.07, and 1.24 (analyzed as 0.58, 0.75, 0.93, 1.11, and 1.30). These five diets were deficient in P but supplemented with 1,000 phytase units/kg feed. Each diet was fed to six pens of eight pigs (four barrows and four gilts per pen). All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen on days 5-7 of trial. At the end, one pig per pen was sacrificed to collect the right tibia and urine in the bladder. The results showed that increasing dietary Ca/P ratio to 0.93 increased gain:feed but then gain:feed decreased as the Ca/P ratio was increased to 1.30 (linear and quadratic, P < 0.05). Although average daily gain and final BW were unaffected by changing Ca/P ratio in diet, dry bone weight; weights of bone ash, Ca and P; and bone Ca/P ratio increased linearly (P < 0.001) with increasing dietary Ca/P ratio. The percent bone Ca showed a tendency to increase (P = 0.064). Increasing dietary Ca/P ratio decreased apparent total tract digestibility (ATTD) of Ca and P linearly (P < 0.05) and the concentration of digestible P linearly (P < 0.001), but increased the concentration of digestible Ca (linear and quadratic effects: P < 0.01) and the digestible Ca/P ratio (linear effect: P < 0.001). In plasma, the concentration of Ca increased both linearly (P < 0.01) and quadratically (P = 0.051), whereas the concentration of P tended (linear and quadratic, P < 0.10) to decrease with increasing dietary Ca/P ratio. Similarly, in urine, the concentration of Ca increased both linearly and quadratically (P < 0.05), whereas the concentration of P decreased linearly (P < 0.01). In conclusion, increasing the dietary Ca/P ratio reduced feed efficiency but increased bone mass and the amounts of Ca and P deposited in bone of nursery pigs fed diets supplemented with 1,000 FYT/kg phytase. The increases in bone growth led to a reduction of urinary P excretion that exceeded the decreased digestible P supplied in diet with the widening dietary Ca/P ratios.

The effect of amylase supplementation on individual variation, growth performance, and starch digestibility in broiler chickens.

The objective of this study was to evaluate the variance of starch digestibility in broilers individually fed diets without or with supplemental exogenous amylase. A total of 120 d-of-hatch male chicks were individually reared from 5 to 42 d in metallic cages and fed maize-based basal diets or diets containing 80 kilo-novo-α-amylase units/kg (60 birds or replicates per treatment). Beginning on d 7, feed intake, body weight gain, and feed conversion ratio were recorded; partial excreta collection was conducted every Monday, Wednesday, and Friday until 42 d, when all birds were sacrificed for individual collection of duodenal and ileal digesta. Lower feed intake (4,675 vs. 4,815 g) and feed conversion ratio (1.470 vs. 1.508) were observed in amylase-fed broilers during the overall period (7-43 d; P < 0.01), whereas body weight gain was not affected. Amylase supplementation improved total tract starch (TTS) digestibility (P < 0.05) on each day of excreta collection (except for d 28, where no difference was found), averaging 0.982 vs. 0.973 compared to basal-fed broilers from d 7 to 42. Both apparent ileal starch (AIS) digestibility and apparent metabolizable energy (AMEN) were increased (P <0.05) from 0.968 to 0.976 and from 3,119 to 3,198 kcal/kg, respectively, with enzyme supplementation. Activity of amylase in the duodenum was higher (18.6 vs. 50.1 IU/g of digesta) in supplemented birds. Amylase supplementation led to a reduced coefficient of variation for both TTS (averaged 2.41 vs. 0.92% from 7 to 42 d) and AIS digestibilities (1.96 vs. 1.03%), as well as AMEN (0.49 vs. 0.35%), when compared to the nonsupplemented group, indicating lower individual heterogenity. An age effect was detected for TTS digestibility, as both groups saw an increase during the first weeks (slightly more pronounced in the supplemented group); older birds (d 30 onwards) presented a lower TTS digestibility compared to ages between 7 and 25 d. In conclusion, amylase supplementation in maize diets for broilers can attenuate individual bird variation for starch and energy utilization by increasing amylase activity and enhancing starch digestibility.

Use of fixed calcium to phosphorus ratios in experimental diets may create bias in phytase efficacy responses in swine.

The objective of this study was to investigate the effects of two dietary total Ca/P ratios on available P release by phytase, measured using growth performance and bone mineralization with 528 barrows and gilts according to a randomized complete block design. Three were 11 diets in a factorial of 2 by 4 plus 3, including 3 reference diets consisting of 0.25% (control), 0.70%, or 1.15% monocalcium phosphate (MCP) and 8 diets from combining 4 phytase doses (500, 1,000, 2,000, and 3,000 FYT/kg) with 0.25% MCP and 2 dietary Ca/P ratios (1.05 and 1.20). Each diet was fed to 6 pens of 8 pigs. All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen on d 13-15 of trial. At the end of trial, one pig per pen was sacrificed to collect a tibia and urine in the bladder. The results showed that MCP improved growth performance linearly (P < 0.01), whereas both a linear and quadratic response was observed with the addition of phytase. The MCP increased the percent bone ash and weights of bone ash, Ca, and P linearly (P < 0.01). At both Ca/P ratios, increasing supplementation of phytase increased the percent bone ash and weights of bone ash, Ca, and P both linearly and quadratically (P < 0.05). Both MCP and phytase significantly increased digestibility of Ca and P as well as digestible Ca and P in diets and reduced the digestible Ca/P ratio. The dietary Ca/P ratio of 1.20 resulted in poorer feed utilization efficiency, more digestible Ca, greater percent bone ash, Ca, and P and heavier weights of bone Ca and P than the ratio of 1.05 (P < 0.05). The ratio of 1.20 elicited numerically higher available P release values from phytase, with percent bone ash and bone P weight as the response variables, but significantly lower values with gain:feed. The urinary concentration of Ca increased linearly (P < 0.01) with increasing digestible Ca/P ratios whilst urinary concentration of P decreased quadratically (P < 0.01). In conclusion, fixing the same total Ca/total P ratio in diets supplemented with increasing phytase dosing created an imbalance of digestible Ca and P, which could have an adverse effect on bone mineralization and thus compromise the phytase efficacy relative to mineral P.

Protease supplementation reduced the heat increment of feed and improved energy and nitrogen partitioning in broilers fed maize-based diets with supplemental phytase and xylanase.

An experiment was conducted to explore the effects of digestible amino acid (dAA) concentrations and supplemental protease on live performance and energy partitioning in broilers. Ross 308 male broilers (n = 288) were distributed into 24 floor pens and offered 1 of 4 dietary treatments with 6 replicates from 1 to 35 d of age. Dietary treatments consisted of a 2 × 2 factorial arrangement with dAA concentrations (standard and reduced [34 g/kg below standard]) and supplemental protease (without or with) as the main factors. At 1, 15, 28, and 35 d of age, feed and broilers were weighed to determine live performance. From 20 to 23 d of age, a total of 32 birds (2 birds/chamber, 4 replicates) were placed in closed-calorimeter chambers to determine respiratory exchange (heat production, HP), apparent metabolisable energy (AME), retained energy (RE), and net energy (NE). From 29 to 35 d of age, supplemental protease in the reduced-dAA diet decreased broiler feed conversion ratio (FCR) by 5.6 points, whereas protease supplementation in the standard-dAA diet increased FCR by 5.8 points. The indirect calorimetry assay revealed that supplemental protease decreased (P < 0.05) the heat increment of feed (HIF) by 0.22 MJ/kg. Also, from 20 to 23 d of age, broilers offered the reduced-dAA diet with supplemental protease had a higher daily body weight gain (BWG) (+10.4%), N intake (+7.1%), and N retention (+8.2%) than those offered the standard-dAA with supplemental protease. Broilers offered the reduced-dAA without supplemental protease exhibited a 3.6% higher AME-to-crude protein (CP) ratio than those offered other treatments. Protease supplementation in the standard- and reduced-dAA diets resulted in 2.7% and 5.6% lower AME intake-to-N retention ratios, respectively, compared with the unsupplemented controls. Reduced-dAA increased (P < 0.05) AME intake (+4.8%), RE (+9.8%), NE intake (+5.8%), NE intake-to-CP ratio (+3.0%), and RE fat-to-RE ratio (+8.6%). Protease supplementation increased (P < 0.05) respiratory quotient (+1.2%) and N retention-to-N intake ratio (+2.2%), NE-to-AME ratio (+1.9%), and reduced HP (-3.6%), heat increment (-7.4%), and NE intake-to-N retention (-2.5%). In conclusion, protease positively affected FCR and energy partitioning in broilers; responses were most apparent in diets with reduced-dAA concentrations.

Dynamic Distribution of Gut Microbiota in Pigs at Different Growth Stages: Composition and Contribution.

Fully understanding the dynamic distribution of the gut microbiota in pigs is essential, as gut microorganisms play a fundamental role in physiological processes, immunity, and the metabolism of nutrients by the host. Here, we first summarize the characteristics and the dynamic shifts in the gut microbial community of pigs at different ages based on the results of 63 peer-review publications. Then a meta-analysis based on the sequences from 16 studies with accession numbers in the GenBank database is conducted to verify the characteristics of the gut microbiota in healthy pigs. A dynamic shift is confirmed in the gut microbiota of pigs at different ages and growth phases. In general, Bacteroides, Escherichia, Clostridium, Lactobacillus, Fusobacterium, and Prevotella are dominant in piglets before weaning, then Prevotella and Aneriacter shift to be the predominant genera with Fusobacterium, Lactobacillus, and Miscellaneous as comparative minors in postweaned pigs. A number of 19 bacterial genera, including Bacteroides, Prevotella, and Lactobacillus can be found in more than 90% of pigs and three enterotypes can be identified in all pigs at different ages, suggesting there is a "core" microbiota in the gut of healthy pigs, which can be a potential target for nutrition or health regulation. The "core" members benefit the growth and gut health of the host. These findings help to define an "optimal" gut microbial profile for assessing, or improving, the performance and health status of pigs at different growth stages. IMPORTANCE The ban on feed antibiotics by more and more countries, and the expected ban on ZnO in feed supplementation from 2022 in the EU, urge researchers and pig producers to search for new alternatives. One possible alternative is to use the so-called "next-generation probiotics (NGPs)" derived from gastrointestinal tract. In this paper, we reveal that a total of 19 "core" bacterial genera including Bacteroides, Prevotella, and Lactobacillus etc., can be found in more than 90% of healthy pigs across different ages. These identified genera may probably be the potential candidates of NGPs or the potential target of microflora regulation. Adding substrates preferred by these target microbes will help to increase the abundance of specific symbiotic species and benefit the gut health of pigs. Further research targeting these "core" microbes and the dynamic distribution of microbiota, as well as the related function is of great importance in swine production.

The Response of Layer Hen Productivity and Egg Quality to an Additional Limestone Source When Offered Diets Differing in Calcium Concentrations and the Inclusion of Phytase.

Laying hens require substantial quantities of calcium (Ca) to maintain egg production. However, maintaining recommended dietary Ca through inclusion of limestone may impede nutrient digestibility, including that of other minerals. It was hypothesized that providing a separate source of dietary Ca in the form of limestone grit would preserve Ca intake of hens offered diets containing suboptimal Ca concentrations. Furthermore, the impact of dietary phytase at a "superdosing" inclusion rate on the voluntary consumption of limestone grit was evaluated. One hundred and forty-four laying hens (19 weeks of age) were assigned to one of six dietary treatments in a 3 × 2 factorial arrangement comprising three dietary Ca concentrations (40, 30, and 20 g/kg) and ±dietary phytase (3500 FYT/kg diet) on an ad libitum basis for six weeks. Limestone grit (3.4 ± 1.0 mm) was provided to all hens ad libitum. Hens offered diets containing phytase consumed significantly less limestone grit p = 0.024). Egg weight, rate of lay, and egg mass were unaffected by dietary treatment (p > 0.05). Egg shell weight % (p < 0.001), shell thickness (p < 0.001), and shell breaking strength (p < 0.01) decreased in line with dietary Ca levels. In summary, dietary superdosing with phytase reduced the consumption of a separate limestone source in individually housed, early lay ISA Brown hens. Egg shell quality variables but not egg production worsened in line with lower dietary Ca levels.

A review of limitations to using cassava meal in poultry diets and the potential role of exogenous microbial enzymes.

Cassava (Manihot esculenta), a crop grown in the tropics, is increasingly becoming a vital feed resource for human beings and livestock. Traditionally, cassava has been used primarily as a source of food for humans. However, it is becoming an increasingly important ingredient in livestock feed. The use of cassava leaves and roots in poultry diets is limited because of nutrient imbalances and toxins (hydrogen cyanide (HCN)) found in them. High HCN is reduced to innocuous levels by processing the ingredient using a simple sun drying method. Plant fibre content can be reduced and made available for use by poultry through the use of exogenous enzymes. More recent innovative interventions in biotechnology have brought about various exogenous enzymes that can help improve the digestibility of fibrous diets. These include, among others, carbohydrases, proteases and phytases. The extent to which the animals utilise nutrients is influenced by the type of enzyme and the physicochemical properties of the feed ingredient. This review aims to collate information on the current state of knowledge on the use of exogenous microbial enzymes in diets containing cassava and how the enzymes that target carbohydrates might be useful in making nutrient available for poultry.

Trends in feed evaluation for poultry with emphasis on in vitro techniques.

Accurate knowledge of the actual nutritional value of individual feed ingredients and complete diets is critical for efficient and sustainable animal production. For this reason, feed evaluation has always been in the forefront of nutritional research. Feed evaluation for poultry involves several approaches that include chemical analysis, table values, prediction equations, near-infrared reflectance spectroscopy, in vivo data and in vitro digestion techniques. Among these, the use of animals (in vivo) is the most valuable to gain information on nutrient utilization and is more predictive of bird performance. However, in vivo methods are expensive, laborious and time-consuming. It is therefore important to establish in vitro methods that are reliable, rapid and practical to assess the nutritional quality of feed ingredients or complete diets. Accuracy of the technique is crucial, as poor prediction will have a negative impact on bird performance and, increase feed cost and environmental issues. In this review, the relevance and importance of feed evaluation in poultry nutrition will be highlighted and the various approaches to evaluate the feed value of feed ingredients or complete diets will be discussed. Trends in and practical limitations encountered in feed evaluation science, with emphasis on in vitro digestion techniques, will be discussed.

Balanced nutrient density for broiler chickens using a range of digestible lysine-to-metabolizable energy ratios and nutrient density: Growth performance, nutrient utilisation and apparent metabolizable energy.

Currently, specific nutrient concentration, metabolizable energy (ME) and digestible amino acids are used as feed formulation criteria. A balanced nutrient density (BND) concept based on 2 criteria of nutrient density and balanced amino acids-to-ME ratio may offer more flexibility in optimisation of profit in formulation of diets compared with current formulation based on set values per unit of feed mass. A total of 672 one-d-old off-sex male Ross 308 broiler chickens were used across two 42-d performance trials in a 3 × 2 factorial arrangement of treatments with each diet replicated 8 times (14 birds per replicate). The experimental factors were 2 nutrient density levels (low [LD] and high [HD]) and 3 digestible lysine-to-ME ratios (DLYS:ME; low, medium, and high). Low density diets had ME of 2,876 and 3,023 kcal/kg for starter and finisher, respectively, while values for HD diets were 3,169 and 3,315 kcal/kg with proportionally higher non-nitrogenated nutrients. Separate digestibility and apparent metabolizable energy (AME) assays were conducted at d 21 and 42. Digestibility assays at d 7 were conducted on birds used for performance trials. Regardless of the diet density, birds fed low DLYS:ME had a lower (P < 0.01) feed intake (d 0 to 42) than medium and high DLYS:ME. Without interaction, birds fed low and medium DLYS:ME had a similar body weight gain being the heaviest while birds low DLYS:ME were the lightest. By an interaction (P < 0.05), the highest overall FCR value was observed for birds fed LD × low DLYS:ME and improved linearly when DLYS:ME increased to the highest level reaching a limit for birds fed HD × medium DLYS:ME. Calorie conversion linearly decreased (P < 0.001) with increments in DLYS:ME. Jejunal and ileal starch and protein digestibility were affected on d 21 and 42 but not on d 7 of age. Given the independence of response on BW and feed consumption, the use of BND as a flexible system in diet formulations has the potential to enable more accurate formulation for optimisation of growth performance of broiler chickens.

The effect of short-term phytase supplementation on the apparent total tract digestibility of calcium and phosphorus and the reproductive performance of late gestation and lactating sows fed diets without mineral phosphorus.

The objective of this study was to measure apparent total tract digestibility (ATTD) of Ca and P as well as reproductive performance in late gestation and lactating sows supplemented with a novel phytase and to compare the response to phytase supplementation between late gestation and lactating sows. A total of 45 late gestation sows and 45 lactating sows were used in experiments 1 and 2, respectively, in a completely randomized design. The sows were provided with a control diet or the control diet supplemented with 187.5 or 375 FYT phytase/kg feed for 10 days. The diets were prepared according to the formulas in use for production but without any inorganic P supplement. Titanium dioxide was included at 3 g/kg feed as an indigestible marker. Each dietary treatment was replicated with 15 sows individually housed in farrowing stalls. The sows were allowed to adapt to the experimental diets for 5 days before a 5-d fecal collection by grab sampling, and the performance of the sows and their litters were measured until weaning. The results showed that the ATTD of Ca increased linearly (P < 0.001), while the ATTD of P increased both linearly and quadratically (P < 0.01) with increasing supplementation of phytase in both late gestation and lactating sows. There was no significant effect of phytase on the ATTD of dry matter, crude protein, and gross energy, and the performance of the sows and their progenies. The phytase added at 187.5 and 375 FYT/kg feed released 0.07% and 0.10% digested P, respectively, in late gestation sows, which compared with 0.09% and 0.12% digested P in lactating sows. In conclusion, a novel phytase at 187.5-375 FYT/kg feed could release 0.07-0.12% digestible P for sows. It appeared that using the P digestibility values of feed ingredients listed by NRC to formulate a diet for sows might overestimate dietary P supply and a greater response to phytase supplementation could be expected in lactating sows than in late gestation sows.

Mathematical prediction of ileal energy and protein digestibility in broilers using multivariate data analysis.

A proportional mixture design was used to systematically create a total of 56 diets using ten feed ingredients. Diets differed widely with regards to chemical characteristics and ingredient inclusion levels. Apparent ileal digestibility of energy and protein of the diets were determined in broiler growers fed ad libitum from 21 to 24 d post-hatch. The chemical composition and the in vivo digestibility values were used to establish prediction equations for energy and protein digestibility, using multivariate data analysis. Root mean square error as percentage of the observed means (RMSE%) and residual error were used to evaluate the strength and accuracy of the predictions and to compare predictions based on chemical characteristics with estimates based on table values. The estimates of ileal digestibility of energy from table values were relatively accurate (RMSE% = 5.15) and was comparable to those predicted based on the chemical composition of diets. Estimates of ileal digestibility of protein based on table values were less accurate (RMSE% = 8.21); however, the prediction was improved by multivariate regression (RMSE% = 5.46) based on chemical composition of diets. The best predictors for ileal energy digestibility were starch, crude fiber and phytate contents (P < 0.01) and the best predictors for crude protein digestibility were starch, CF and fat contents (P < 0.05). In conclusion, the ileal digestibility of energy can be accurately predicted using table values; however, the accuracy of prediction of the ileal digestibility of protein can be improved when chemical characteristics of the diet are considered.

Latent Anti-nutrients and Unintentional Breeding Consequences in Australian Sorghum bicolor Varieties.

Modern feed quality sorghum grain has been bred to reduce anti-nutrients, most conspicuously condensed tannins, but its inclusion in the diets of monogastric animals can still result in variable performance that is only partially understood. Sorghum grain contains several negative intrinsic factors, including non-tannin phenolics and polyphenols, phytate, and kafirin protein, which may be responsible for these muted feed performances. To better understand the non-tannin phenolic and polyphenolic metabolites that may have negative effects on nutritional parameters, the chemical composition of sorghum grain polyphenol extracts from three commercial varieties (MR-Buster, Cracka, and Liberty) was determined through the use of an under-studied, alternative analytical approach involving Fourier-transform infrared (FT-IR) spectroscopy and direct ionization mass spectrometry. Supervised analyses and interrogation of the data contributing to variation resulted in the identification of a variety of metabolites, including established polyphenols, lignin-like anti-nutrients, and complex sugars, as well as high levels of fatty acids which could contribute to nutritional variation and underperformance in monogastrics. FT-IR and mass spectrometry could both discriminate among the different sorghum varieties indicating that FT-IR, rather than more sophisticated chromatographic and mass spectrometric methods, could be incorporated into quality control applications.

The influence of feed ingredients on CP and starch disappearance rate in complex diets for broiler chickens.

The influence of feed ingredients on digestion kinetics of N and starch in complex diets was investigated in the current experiment. A total of 34 diets with different inclusion levels of 10 commonly used feed ingredients (corn, wheat, sorghum, soybean meal, canola meal, full-fat soybean meal [FFSB], palm kernel meal, meat and bone meal, wheat distillers grain with solubles and wheat bran) were randomly allocated to 170 cages with 8 birds in each. Apparent jejunal and ileal digestibility of N and starch was determined on a cage level in broilers feed the experimental diets ad libitum from 21 to 24 d after hatch. Disappearance rate of N and starch from the intestine was estimated through a first-order decay function fitted to the digesta data from the jejunum and ileum. The fit of the decay functions was evaluated with root mean squared error as percentage of the observed mean. The influence of the feed ingredients on the disappearance rates were found through a linear regression model, including the effect of the single ingredients, 2-way and 3-way interactions and evaluated with a Student t test test. Starch digestion kinetics were in general faster than N digestion kinetics. The N disappearance rate was both influenced by single ingredients and interaction amongst ingredients, whereas starch disappearance rate mainly was influenced by single ingredients. A combination of FFSB and soybean meal decreased the N digestion rate by 22 to 25% compared with diets with only soybean meal or FFSB, respectively. These results indicate that nutrients from 1 feed ingredient can influence the rate of disappearance of nutrients from other feed ingredients in a complex diet. This highlights the importance of understanding nutrient digestion kinetics and how these are influenced both additively and nonadditively by different feed ingredients in complex diets.

Corn drying temperature, particle size, and amylase supplementation influence growth performance, digestive tract development, and nutrient utilization of broilers.

Broiler live performance may be influenced by postharvest corn drying temperature, and results could depend on particle size after grinding. The supplementation with an exogenous amylase may improve performance parameters, but responses to enzymes are also affected by particle size. Two parallel experiments were conducted to evaluate the effects of hard-kernel corn dried at 2 temperatures (35°C and 120°C), ground at 2 particle sizes (coarse or fine), and 3 supplementation levels (0, 133, and 266 g ton-1) of an exogenous amylase on live performance, gastrointestinal organ development, energy utilization, and nutrient digestibility. Twelve dietary treatments resulting from a 2 × 2 × 3 factorial arrangement of drying temperature, particle size, and amylase supplementation were evaluated in both experiments. A total of 1,920 day-old male chicks were randomly allocated to 96 floor pens, while 480 chicks were distributed among 4 battery brooder units. Ileal and fecal samples were collected to determine energy utilization and nutrient digestibility using titanium dioxide as inert marker. At 42 D, organs were collected, and relative weight or length was determined. Data were analyzed using a three-way ANOVA in a randomized complete block design. Feeding fine corn-based diets showed improvements on live performance for both studies. At 40 D, supplementing 266 g ton-1 of amylase improved feed conversion ratio (P < 0.05) by 1 point compared to chickens that consumed nonsupplemented diets and feed with amylase at 133 g ton-1. Broilers fed coarse corn-based diets had heavier gizzard (P < 0.001) and liver (P < 0.05) than chickens that consumed fine corn-based diets. In addition, starch digestibility was improved by amylase (P < 0.05) at 133 g ton-1 and by feeding coarse corn-based diets (P = 0.06). For chicks raised in cages (16 D), AMEn was increased (P < 0.01) by amylase supplementation regardless of its inclusion level. In conclusion, drying temperature and particle size interactions influenced broiler live performance, gastrointestinal organ development, nutrient digestibility, and energy utilization, and these parameters were improved by supplementing amylase.

Soybean meal allergenic protein degradation and gut health of piglets fed protease-supplemented diets.

Two experiments were conducted to determine the effects of protease supplementation on degradation of soybean meal (SBM) allergenic proteins (glycinin and ß-conglycinin) and gut health of weaned pigs fed soybean meal-based diets. In experiment 1, 2 SBM samples from 2 different sources were subjected to porcine in vitro gastric degradation to determine the effects of protease (at 15,000 U/kg of feedstuff) on degradation of the soybean allergenic proteins. In experiment 2, 48 weaned pigs (body weight = 6.66 kg) were obtained in 2 batches of 24 pigs each. Pigs were individually housed in metabolic crates and fed 4 diets (12 pigs/diet). The diets were corn-based diet with SBM 1 or SBM 2 without or with protease at 15,000 U/kg of diet in 2 × 2 factorial arrangement. Diets were fed for 10 d and pigs were sacrificed on day 10 for measurement of small intestinal histomorphology, permeability of small intestine mounted in Ussing chambers, and serum concentration of pro-inflammatory cytokines. Two SBM sources (SBM 1 and SBM 2) contained 46.9% or 47.7% CP, 14.0% or 14.6% glycinin, and 9.90% or 10.3% ß-conglycinin, respectively. Protease and SBM source did not interact on any of the response criteria measured in the current study. Protease supplementation tended to increase (P = 0.069) the in vitro gastric degradation of glycinin. Protease supplementation tended to reduce (P = 0.099) fluorescein isothiocyanate dextran 4,000 Da (which is a marker probe for intestinal permeability) flow in jejunum, and reduced (P = 0.037) serum TNF-α concentration. Protease did not affect small intestinal histomorphology. In conclusion, protease tended to increase gastric degradation of glycinin and reduce gut permeability, and serum concentration of pro-inflammatory cytokines, indicating that the protease used in the current study can be added to SBM-based diets for weanling pigs to improve gut health.

Phytase catalysis of dephosphorylation studied using isothermal titration calorimetry and electrospray ionization time-of-flight mass spectroscopy.

Phytases are important commercial enzymes that catalyze the dephosphorylation of myo-inositol hexakisphosphate (phytate) to its lower inositol phosphate (IP) esters, IP6 to IP1. Digestion of phytate by Citrobacter braakii 6-phytase deviates significantly from monophasic Michaelis-Menten kinetics. Analysis of phytate digestion using isothermal titration calorimetry (ITC) using the single injection method produced a thermogram with two peaks consistent with two periods of high enzyme activity. Continuous-flow electrospray ionization time-of-flight mass spectroscopy (ESI-ToF-MS) provided real-time analysis of phytase catalysis. It was able to show that the first two cleavage steps were rapid and concurrent but the third cleavage step from IP4 to IP3 was slow. The third (IP4 to IP3), fourth (IP3 to IP2) and fifth (IP2 to IP1) cleavages were effectively sequential due to the preferred association of the more phosphorylated species with the phytase catalytic site. This created a bottleneck during the cleavage of IP4 to IP3 until the point at which IP4 was exhausted and was followed by the rapid cleavage of IP3 to IP2, which was observed as the second peak in the ITC thermogram. This work illustrates the importance of an orthogonal approach when studying non-specific or complex enzyme catalyzed reactions.

Monitoring Phytate Hydrolysis Using Serial Blood Sampling and Feather Myo-Inositol Levels in Broilers.

Phytate forms insoluble precipitates with various cations that are recalcitrant to digestion in poultry. Dietary supplementation with exogenous phytase has been shown to improve phytate solubility and digestibility and, in turn, improve animal growth performance. Although the kinetics of phytate hydrolysis by exogenous phytase are well described in vitro, the progression of the reaction in vivo is still not well defined. The aim of the present study was, therefore, to monitor the kinetic variation of myo-inositol (myo-Ins) levels in both circulation and feather following exogenous phytase supplementation. In experiment 1, 4 week-old male broilers were individually housed with ad libitum access to water and a standard commercial diet. Birds were maintained under environmental temperature of 24°C and 30% RH. Birds were cannulated in the cutaneous ulnar vein on the right wing and remained untouched for 3 days. On the day of the experiment, birds were randomly divided into three body weight-matched groups and fed either the control diet, the control diet-supplemented with myo-Ins or Ronozyme HiPhos (0.06%, DSM Nutritional Products, Switzerland) for 10 h. In the experiment 2, birds were fed only HiPhos for 30 h. Growing feathers and blood were collected at baseline and then every 2 h for 10 h (experiment 1) and 30 h (experiment 2) post-prandially. Plasma and feather myo-Ins levels were determined by UHPLC-MS/MS. The relative expression of inositol polyphosphate-1-phosphatase (INPP1), inositol hexakisphosphate kinase 1-3 (IP6K1-3), inositol-3-phosphate synthase (ISYNA), and multiple inositol-polyphosphate phosphatase 1 (MNPP1) genes in blood and feathers was determined by real-time qPCR using 2-ΔΔCt method. Plasma and feather myo-Ins levels were significantly increased by HiPhos at 6 h to 8 h post-prandial. The mRNA abundances of INPP1, IP6K1, and ISYNA in the circulation were significantly down regulated at all periods compared to the baseline levels. IP6K2, IP6K3, and MINPP1 gene expression, however, was up regulated at 8 h post-prandial and then returned to the baseline levels. In feathers, the expression of INPP1 was induced at 8 h post-prandial and remained higher compared to the baseline. The expression of IP6K2, IP6K3, and MINPP1 was down regulated during the first 10 h and then returned to baseline levels for the rest of the post-prandial period. Taken together, our data show that phytase modulates the expression of genes associated with myo-Ins metabolism and generates release of myo-Ins in both circulation and feather at 6-10 h post-feeding. Feather myo-Ins concentration could be used as a non-invasive method to monitor phytate hydrolysis in practice.

Digestibility of amino acids, energy, and minerals in roasted full-fat soybean and expelled-extruded soybean meal fed to growing pigs without or with multienzyme supplement containing fiber-degrading enzymes, protease, and phytase.

Indigestible fiber-protein-phytate complexes reduce the feeding value of soy products. We investigated the effects of multienzyme supplement (MES, Victus) on standardized ileal digestibility (SID) of amino acids (AA) and apparent total tract digestibility (ATTD) of energy and minerals in roasted full-fat soybean (FFSB) seeds and expelled-extruded soybean meal (SBM) fed to growing pigs. The crude protein (CP) was 33.4% and 42.8% dry matter (DM) in FFSB seeds and SBM, respectively and corresponding values for crude fat were 17.4% and 11.8% DM. Semi-purified diets with 50% of either FFSB seeds or SBM as the sole source of AA were prepared without or with MES supplying phytase, protease, xylanase, and ß-glucanase at 2,200, 8,300, 400, and 100 U/kg of feed, respectively. Diets had TiO2 as an indigestible marker and the ratio of cornstarch to sucrose and corn oil was identical to calculate DE by the difference method. Eight ileal-cannulated barrows (22.1 ± 0.61 kg) were fed diets in a replicated 4 × 4 Latin square design to give eight replicates per diet. The period lasted for 9 d: 5 d for acclimation, 2 d for fecal, and 2 d for ileal digesta samples. There was no (P > 0.05) interaction between soy type and MES or MES effect on SID of AA; SBM had higher (P < 0.05) SID of CP, His, Leu, and Lys. There was no (P > 0.05) interaction between soy type and MES on energy digestibility. The FFSB seeds had higher ATTD of gross energy (GE, 80.2% vs. 76.6%; P < 0.01) than SBM. Pigs fed MES had higher (P < 0.05) ATTD of DM (91.3% vs. 87.7 %), GE (87.5% vs. 82.4%), CP (86.4% vs. 82.9%), crude fat (70.6% vs. 54.9%), Ca (63.2% vs. 60.2%), and P (67.5% vs. 63.2%). In conclusions, differences on AA and energy digestibility in soy products could be linked to processing and compositional differences. Although MES had no effect on SID of AA, the effects on the utilization of minerals and energy demonstrated the value of fiber-degrading enzymes, protease, and phytase in improving the nutritive value of soy products independent of processing.