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Thirteen pea grain samples from different origins were used to examine the variation in in situ ruminal degradation of crude protein (CP) and starch as well as in vitro gas production (GP) kinetics. In vitro GP was used to estimate the digestibility of organic matter (dOM), metabolisable energy (ME) and utilisable CP at the duodenum (uCP). Protein fractions were also determined according to the Cornell Net Carbohydrate and Protein System. Degradation of CP and starch from all pea grains in the rumen was almost complete, with a high proportion of the instantly disappearing fraction. The variation in the degradation constants among pea grain variants was high, and degradation of CP and starch showed a significant initial lag phase in the rumen. The mean effective degradation (ED) calculated for a rumen outflow of k = 8%/h of CP (EDCP8) was higher than ED of starch (EDST8), averaging 77.0 and 71.5%, respectively, with low variation among variants. A correlation analysis between GP parameters and in situ degradation constants showed no clear relationship, but the rates of in vitro GP and in situ starch degradation were similar. Most of the protein in the pea grains was buffer-soluble with fast and intermediate degradation. Variation in the protein fractions among the pea grain variants was low and not suitable for predicting differences in in situ degradation characteristics. The mean in vitro uCP of pea grains was 198 g/kg dry matter (k = 8%/h) and variation was low and consistent with that of GP kinetics and in situ rumen undegradable crude protein values. The estimation of dOM and ME from 24 h GP led to very high values indicating that the existing prediction equations may not be suitable for pea grains as a single feed.

Feed and phosphorus (P) efficiency are of increasing importance in poultry breeding. It has been shown recently that these efficiency traits are influenced by the gut microbiota composition of the birds. The efficiency traits and the gut microbiota composition are partly under control of the host genome. Thus, the gut microbiota composition can be seen as a mediator trait between the host genome and the efficiency traits. The present study used data from 749 individuals of a Japanese quail F2 cross. The birds were genotyped for 4k single-nucleotide polymorphism (SNP) and trait recorded for P utilization (PU) and P retention (PR), body weight gain (BWG), and feed per gain ratio (F:G). The gut microbiota composition was characterized by targeted amplicon sequencing. The alpha diversity was calculated as the Pielou’s evenness index (Jʹ). A stable Bayesian network was established using a Hill-Climbing learning algorithm. Pielou’s evenness index was placed as the most upstream trait and BWG as the most downstream trait, with direct and indirect links via PR, PU, and F:G. The direct and indirect effects between Jʹ, PU, and PR were quantified with structural equation models (SEM), which revealed a causal link from Jʹ to PU and from PU to PR. Quantitative trait loci (QTL) linkage mapping revealed three genome-wide significant QTL regions for these traits with in total 49 trait-associated SNP within the QTL regions. SEM association mapping separated the total SNP effect for a trait into a direct effect and indirect effects mediated by upstream traits. Although the indirect effects were in general small, they contributed to the total SNP effect in some cases. This enabled us to detect some shared genetic effects. The method applied allows for the detection of shared genetic architecture of quantitative traits and microbiota compositions.

1. The objective of this study was to investigate wheat genotypes bred for increased intrinsic phytase activity for InsP6 disappearance and the formation of lower inositol phosphates in such wheat-fed broiler chickens. The influence of monocalcium phosphate (MCP) supplementation on these characteristics and the utilisation of P and Ca were also determined. A three-step in vitro assay and a broiler trial were performed.2. In the 63 wheat genotypes tested in vitro, phytase activity varied from 1900 FTU/kg to 5200 FTU/kg, and InsP6 disappearance increased with higher phytase activity of wheat in a linear manner. The addition of MCP significantly reduced in vitro InsP6 disappearance by one-third, independent of the inclusion level of wheat in the feed. When exogenous phytase was added to wheat, in vitro InsP6 disappearance increased independently of the phytase activity of the wheat used.3. In the broiler trial, four wheat genotypes with phytase activities between 2400 and 3700 FTU/kg were included at 400 g/kg in diets with and without MCP. The diets were not pelleted. Separately, wheat 1, without MCP, was tested with the addition of exogenous phytase. Unsexed Ross 308 broiler chickens were allocated to 72 metabolic units of 10 birds each and assigned one of the nine diets. Mineral utilisation was measured based on excreta collection from 20 to 23 d of age. Digesta from the crop and terminal ileum were collected on d 24.4. In the crop and ileum, InsP6 disappearance was not affected by the wheat genotypes, but the addition of MCP significantly decreased InsP6 disappearance. Precaecal P disappearance was significantly reduced by the addition of MCP, with wheat genotypes also exerting an effect. Wheat genotypes and the addition of exogenous phytase significantly affected P utilisation. Exogenous phytase had no effect on InsP6 disappearance in the crop but did up to the terminal ileum, the precaecal InsP6 and P disappearance increased with the addition of exogenous phytase.5. Although the intrinsic wheat phytase activity exerted distinct effects on in vitro InsP6 disappearance, no such effect was found in the broiler trial. The addition of MCP significantly inhibited InsP6 degradation in vitro and in vivo.

The objective of this contribution was to summarize from scientific literature the optimal concentration of nonphytate phosphorus (NPP) in feed for laying hens. The considered studies were one meta-analysis from 2012 and original studies published since then. Dietary treatments in the studies included variation in supplementation with mineral P sources and phytase. The studies investigated different periods of production and varied in duration but data were insufficient to analyze such factors in a systematic way. No study showed a positive effect on performance and eggshell when the NPP concentration was increased above 2.2 g NPP/kg of feed without the use of phytase. At such level, no consistent impairment of various bone quality traits were found but only few studies on bone quality traits were published. Overall, the data suggested that not more than 2.2 g NPP/kg of feed is needed for laying hens in different stages of production. This value can be reduced when phytase is added to the feed. Such reduction may differ depending on factors such as phytate content of the feed and phytase dosage. However, data are insufficient for calculating precise values of reduction. While phytate degradation in laying hens was markedly increased by phytase supplementation in several studies, effects of phytase supplementation on performance and bone traits in laying hens were less conclusive probably because the hens were supplied more than their NPP requirement. Transition to a system based on digestible P for laying hens similar to broiler chickens may support more precise P nutrition and more sustainable egg production in the future.

Aggregation of data, including deep sequencing of mRNA and miRNA data in jejunum mucosa, abundance of immune cells, metabolites, or hormones in blood, composition of microbiota in digesta and duodenal mucosa, and production traits collected along the lifespan, provides a comprehensive picture of lifelong adaptation processes. Here, respective data from two laying hen strains (Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL) collected at 10, 16, 24, 30, and 60 wk of age were analyzed. Data integration revealed strain- and stage-specific biosignatures, including elements indicative of molecular pathways discriminating the strains. Although the strains performed the same, they differed in the activity of immunological and metabolic functions and pathways and showed specific gut-microbiota-interactions in different production periods. The study shows that both strains employ different strategies to acquire and maintain their capabilities under high performance conditions, especially during the transition phase. Furthermore, the study demonstrates the capacity of such integrative analyses to elucidate molecular pathways that reflect functional biodiversity. The bioinformatic reduction of the multidimensional data provides good guidance for further manual review of the data.

The objective of this study was to compare 2 laying hen strains in 5 production periods regarding phytase activity, phytate (InsP6) degradation, and myo-inositol (MI) release in the digestive tract and phosphorus (P) and calcium (Ca) utilization. One offspring of 10 nonrelated roosters per strain (Lohmann Brown-classic (LB) or Lohmann LSL-classic (LSL)) was placed in one of 20 metabolic units in a completely randomized block design in week 8, 14, 22, 28, and 58 of life. All hens were fed the same corn and soybean meal–based diet at one time, but the diet composition was adjusted to the requirements in the respective period. For 4 consecutive days, excreta were collected quantitatively at 24-hour intervals. In week 10, 16, 24, 30, and 60, the blood plasma, digesta of crop, gizzard, jejunum, ileum, and ceca, and mucosa of the jejunum was collected. The concentration of inorganic P in the blood plasma was higher in LB than in LSL hens (P = 0.026). Plasma Ca concentrations increased with each period (P

As a constituent of animal cells, myo-inositol (MI) has been hypothesized to be crucial in several metabolic and regulatory pathways. Recently, it was shown that dietary phytase contributes to release of MI from phytate in the poultry digestive tract, increasing its systemic concentrations. This study investigated the activities of phosphatases in the jejunum and systemic plasma MI concentration in broilers not supplemented or supplemented with phytase through analyses based on modifications from commercial enzyme activity kits. Three hundred sixty male Ross 308 broilers were randomly allocated to 24 pens (15 birds per pen) in 4 dietary groups. The positive control group was fed with an adequate basal diet. The negative control group (NC) was fed with a reduced level of P and Ca. Groups Phy1500 and Phy3000 were fed with the NC diet plus 1,500 or 3,000 FTU of phytase per kilogram of feed, respectively. One bird per pen was selected for the measurement of jejunal phosphatase activity; MI concentration in plasma, the liver, and the kidney; and key MI enzyme concentrations (liver inositol monophosphatase 1 [IMPase 1] and kidney myo-inositol oxygenase [MIOX]). Endogenous phytase and alkaline phosphatase activity as well as IMPase 1 and MIOX expression were not statistically different among the dietary groups. The supplementation of 1500 FTU of phytase per kilogram of feed resulted in increase of plasma (P < 0.001) and kidney (P < 0.05) but not liver MI concentrations. The results indicated that systemic MI might reflect MI released from dietary sources; however, it did not appear to change expression of enzymes related to endogenous MI synthesis in the liver and catabolism in the kidney. New and larger studies are necessary to reach stronger evidence on the effects of dietary phytase on intestinal and systemic MI concentrations in broilers.

Reducing dietary CP for broiler chickens below a certain threshold results in decreased growth, even when the supply of essential amino acids and glycine equivalent (Glyequi) is adequate, probably because other nonessential amino acids (neAA) are growth-limiting. Nonprotein nitrogen (NPN) might be used for the synthesis of neAA. Therefore, the effects of specific neAA and ammonium chloride (NH4Cl) supplementation on the growth and N-excretion characteristics of broiler chickens were investigated. Nine male Ross 308 broiler chickens were kept in each of 81 metabolism units from day 7 to 21 and received 1 of 9 diets in 9 replicates in a one-factorial arrangement of treatments. Two diets with different neAA concentrations, except for Glyequi, were mixed resulting in CP levels of 180 (CP180) and 160 (CP160) g/kg. In six other diets, CP160 was supplemented with either l-Ala, l-Pro, l-Asp, a mix of l-Asp and l-Asn·H2O, l-Glu, or a mix of l-Glu and l-Gln to achieve concentrations of the respective neAA as formulated in CP180. In a further diet, NH4Cl was added to CP160 to achieve the CP concentration of CP180. The ADG and gain:feed ratio (G:F) from day 7 to 21 were highest at CP180. Reduced neAA concentrations in CP160 decreased ADG and G:F. Supplementation of Asp+Asn, Glu, and Glu+Gln to CP160 increased ADG and G:F, but not to the level found for CP180. Compared with CP160, addition of Asp increased G:F but not ADG. Supplementation of Asp+Asn caused higher ADG and G:F than supplementation of Asp alone. The N-utilization efficiency was highest at CP160 and at CP160 supplemented with Ala, Pro, and Glu. Lower N-utilization efficiency was found at CP180 than at CP160, without and with supplemented neAA. The treatment containing NH4Cl presented the lowest ADG, G:F, and N-utilization efficiency. These results showed that individual supplementation of Asp+Asn, Glu, and Glu+Gln partly compensates for the growth-reducing effects of very low CP diets. Supplementation of NH4Cl as NPN source is not suitable for broiler chickens.

The objective was to determine phytase effects on prececal amino acid (AA) digestibility and phytate (InsP6) breakdown when different oilseed meals were used in broiler chicken diets. The study included 14 diets: a corn-soybean meal (SBM) basal diet and 6 diets that contained SBM, rapeseed meal (RSM), and sunflower meal (SFM) with 2 inclusion levels at the expense of corn starch (150 and 300 g/kg SBM or SFM, or 100 and 200 g/kg RSM). Each diet was mixed with or without a phytase supplement of 1,500 FTU/kg. Diets were provided to broilers for 5 D. Digesta from the posterior half of the ileum were collected on day 21. The average essential AA digestibility, calculated by a regression approach, without and with phytase was 84 and 85% (SBM), 74 and 77% (SFM), and 66 and 73% (RSM), respectively. In the diets, phytase effects on AA digestibility were lower owing to other protein sources also present in the diet, but significant. Prececal InsP6 disappearance was significantly affected by interactions between oilseed meal, inclusion level, and phytase supplementation. Overall, prececal InsP6 disappearance was higher in SBM diets (52%) than in SFM diets (38%) and intermediate in RSM diets (43%). Across diets, phytase supplementation effects on prececal InsP6 degradation linearly increased with the InsP6 concentration of the diet up to 12 g/kg DM. The only exception from linearity was the diet with the high inclusion of SFM, which contained 15.9 g InsP6/kg DM. In the ileal content, the concentration of myo-inositol was significantly increased by phytase supplementation, and this effect was highest in the diets that contained SBM as the only oilseed meal. Concentrations of lower inositol phosphates were increased by phytase supplementation, and this effect was most remarkable for Ins(1,2,3,4)P4 and inositol tetrakisphosphates. The study showed that phytase effects on AA digestibility varied among the 3 tested oilseed meals, but these differences were not detectable in the diets containing these meals. Although phytase effects on ileal content of InsP6 and its degradation products were substantial, they were not related to the effects on AA digestibility.

Myo-inositol (MI) has gained relevance in physiology research during the last decade. As a constituent of animal cells, MI was proven to be crucial in several metabolic and regulatory processes. Myo-inositol is involved in lipid signaling, osmolarity, glucose, and insulin metabolism. In humans and rodents, dietary MI was assessed to be important for health so that MI supplementation appeared to be a valuable alternative for treatment of several diseases as well as for improvements in metabolic performance. In poultry, there is a lack of evidence not only related to specific species-linked metabolic processes but also about the effects of dietary MI on performance and health. This review intends to provide information about the meaning of dietary MI in animal metabolism as well as to discuss potential implications of dietary MI in poultry health and performance with the aim to identify open questions in poultry research.

Background Phosphorus is an essential nutrient in all living organisms and, currently, it is the focus of much attention due to its global scarcity, the environmental impact of phosphorus from excreta, and its low digestibility due to its storage in the form of phytates in plants. In poultry, phosphorus utilization is influenced by composition of the ileum microbiota and host genetics. In our study, we analyzed the impact of host genetics on composition of the ileum microbiota and the relationship of the relative abundance of ileal bacterial genera with phosphorus utilization and related quantitative traits in Japanese quail. An F2 cross of 758 quails was genotyped with 4k genome-wide single nucleotide polymorphisms (SNPs) and composition of the ileum microbiota was characterized using target amplicon sequencing. Heritabilities of the relative abundance of bacterial genera were estimated and quantitative trait locus (QTL) linkage mapping for the host was conducted for the heritable genera. Phenotypic and genetic correlations and recursive relationships between bacterial genera and quantitative traits were estimated using structural equation models. A genomic best linear unbiased prediction (GBLUP) and microbial (M)BLUP hologenomic selection approach was applied to assess the feasibility of breeding for improved phosphorus utilization based on the host genome and the heritable part of composition of the ileum microbiota. Results Among the 59 bacterial genera examined, 24 showed a significant heritability (nominal p ≤ 0.05), ranging from 0.04 to 0.17. For these genera, six genome-wide significant QTL were mapped. Significant recursive effects were found, which support the indirect host genetic effects on the host’s quantitative traits via microbiota composition in the ileum of quail. Cross-validated microbial and genomic prediction accuracies confirmed the strong impact of microbial composition and host genetics on the host’s quantitative traits, as the GBLUP accuracies based on the heritable microbiota-mediated components of the traits were similar to the accuracies of conventional GBLUP based on genome-wide SNPs. Conclusions Our results revealed a significant effect of host genetics on composition of the ileal microbiota and confirmed that host genetics and composition of the ileum microbiota have an impact on the host’s quantitative traits. This offers the possibility to breed for improved phosphorus utilization based on the host genome and the heritable part of composition of the ileum microbiota.

Wheat is of high importance for a healthy and sustainable diet for the growing world population, partly due to its high mineral content. However, several minerals are bound in a phytate complex in the grain and unavailable to humans. We performed a series of trials to compare the contents of minerals and phytic acid as well as phytase activity in several varieties from alternative wheat species spelt, emmer and einkorn with common wheat. Additionally, we investigated the potential of recent popular bread making recipes in German bakeries to reduce phytic acid content, and thus increase mineral bioavailability in bread. For all studied ingredients, we found considerable variance both between varieties within a species and across wheat species. For example, whole grain flours, particularly from emmer and einkorn, appear to have higher mineral content than common wheat, but also a higher phytic acid content with similar phytase activity. Bread making recipes had a greater effect on phytic acid content in the final bread than the choice of species for whole grain flour production. Recipes with long yeast proofing or sourdough and the use of whole grain rye flour in a mixed wheat bread minimized the phytic acid content in the bread. Consequently, optimizing food to better nourish a growing world requires close collaboration between research organizations and practical stakeholders ensuring a streamlined sustainable process from farm to fork.

During life, the number and function of immune cells change with potential consequences for immunocompetence of an organism. In laying hens, studies have primarily focused on early development of immune competence and only few have investigated systemic and lymphatic distribution of leukocyte subsets during adolescence and the egg-laying period. The present study determined the number of various leukocyte types in blood, spleen, and cecal tonsils of 10 Lohmann Brown-Classic and 10 Lohmann LSL-Classic hens per wk of life 9/10, 15/16, 23/24, 29/30, and 59/60, encompassing important production as well as developmental stages, by flow cytometry. Although immune traits differed between the 2 hen strains, identical patterns of age-related immunological changes were found. The numbers of all investigated lymphocyte types in the spleen as well as the numbers of blood γδ T cells increased from wk 9/10 to 15/16. This suggests an ongoing release of lymphocytes from primary lymphoid tissues and an influx of blood lymphocytes into the spleen due to novel pathogen encounters during adolescence. A strong decrease in the number of CTL and γδ T cells and an increase in innate immune cells within blood and spleen were found between wk of life 15/16 and 23/24, covering the transition phase to egg-laying activity. Numbers of peripheral and splenic lymphocytes remained low during the egg-laying period or even further decreased, for example blood CD4+ T cells and splenic γδ T cells. Functional assessments showed that in vitro IFN-γ production of mitogen-stimulated splenocytes was lower in wk 60. Taken together, egg-laying activity seems to alter the immune system toward a more pronounced humoral and innate immune response, with probable consequences for the immunocompetence and thus for productivity, health and welfare of the hens.

Most traits in animal breeding, including feed efficiency traits in pigs, are affected by many genes with small effect and have a moderately high heritability between 0.1 and 0.5, which enables efficient selection. Since the microbiota composition in the gastrointestinal tract is also partly heritable and was shown to have a substantial effect on feed efficiency, the host genes affect the phenotype not only directly by altering metabolic pathways, but also indirectly by changing the microbiota composition. The effect m i of the microbiota composition on the breeding value g i of an animal i is the conditional expectation of its breeding value, given the vector φ i with microbiota frequencies, that is m i = E g i | φ i . The breeding value g i of an animal can therefore be decomposed into a heritable contribution m i that arises from an altered microbiota composition and a heritable contribution p i that arises from altered metabolic pathways within the animal, so g i = m i + p i . Instead of selecting for breeding value g ^ i , an index comprising the two components m ^ i and p ^ i with appropriate weights, that is I i = λ 1 m ^ i + λ 2 p ^ i , can be used. The present study shows how this breeding strategy can be applied in pig genomic selection breeding scheme for two feed efficiency traits and daily gain.

The purpose of this study was to investigate the effects of phytase and protease supplementation on prececal (pc) amino acid (AA) digestibility, phytate (InsP6) degradation, and MEn concentration in diets using 3 oilseed meals as main protein sources in broiler chicken feed. The broiler chicken diets, which lacked mineral phosphorus, contained either soybean meal (SBM), SBM and rapeseed meal (SBM/RSM), or SBM and sunflower meal (SBM/SFM) as main protein sources. Diets were not supplemented with enzymes or supplemented with 1,500 or 3,000 FTU phytase/kg, or with 1,600mg protease/kg. For diets containing SBM as the main protein source, the effects of phytase supplementation with and without monocalcium phosphate were also investigated. Data were obtained during 2 subsequent runs from days 14 to 22 and from days 23 to 31. Each diet was tested using 8 replicates with 4 replicates per run. For pc AA digestibility, no significant interactions were observed between main protein sources, enzyme supplementation, or addition of monocalcium phosphate except for Cys. Supplementation of 1,500 FTU phytase/kg increased pc digestibility of all AA. No differences in pc AA digestibility were observed between 1,500 and 3,000 FTU phytase/kg supplementation treatments. Prececal disappearance of InsP6 and pc P digestibility were greater in the high phytase supplementation treatment. Protease supplementation increased pc digestibility of all AA except for Cys when SBM/RSM was the main protein source. Supplementation of protease and 3,000 FTU phytase/kg increased MEn concentrations. The effect of phytase on pc AA digestibility was fully expressed at a lower supplementation level than needed for a maximized pc InsP6 disappearance and MEn concentration.

The main objective of this study was to perform quantitative genetic analyses of tibia and foot ash traits, which might serve as proxy traits to improve phosphorus utilization (PU) in a breeding program. Additionally, data for ash concentration in tibia and foot were compared with data for total amount of ash. Heritabilities for bone ash traits and genetic and phenotypic correlations between bone ash traits and PU were estimated. A total of 887 F2 birds, established from 2 Japanese quail lines divergently selected on social reinstatement behavior, were provided a P deficient diet. In a metabolic study, feed consumption was measured and total excreta collected for each bird separately. Afterwards, birds were euthanized, the bones obtained and incinerated. Bone ash data showed a heritability of 0.230 (amount of tibia ash) to 0.342 (amount of foot ash), which was higher than estimated for PU, P retention, calcium utilization (0.120–0.174), and performance traits (0.088–0.114). The strongest genetic and phenotypic correlations between PU and bone ash traits were detected for the amount of foot ash with 0.549 and 0.527, respectively. Genetic and phenotypic correlations were stronger between PU and ash amount than between PU and ash percentage, irrespective of bone. Therefore, ash amount was considered a better trait than ash percentage to reflect PU. Strong genetic and phenotypic correlations were detected between the amount of foot and tibia ash (0.887 and 0.901, respectively). Phenotypic and genetic correlations between ash amount and PU or calcium utilization were almost identical, irrespective of bone. Foot ash is as suitable as tibia ash, but easier to determine. Bone ash data, especially the amount of foot ash, seem to be suitable indirect selection criteria for P efficiency breeding.

Dairy cows are commonly fed compound feed concentrates, whose accurate formulation relies on the additivity of ruminal degradation characteristics of single feeds, and the absence of associative effects. The main aim of this study was to evaluate the additivity of single feeds in compound feeds made thereof. Twelve single feeds were used to produce eight compound feeds in mash and pelleted form. Samples of single and compound feeds were incubated in situ in three ruminally fistulated dairy cows, and effective ruminal degradation (ED) of CP and starch (ST) was computed. The ED values of examined compound feeds could be, in most cases, accurately calculated from ED values of single feeds. Observed EDCP values were significantly lower than that calculated, but differences were overall small and not exceeded 5% points. No significant differences were observed between calculated and observed EDST. The study also examined the effects of pelleting of compound feeds on in situ degradation. Pelleting significantly increased EDCP (up to 8% points), and EDST (up to 4% points) of most compound feeds. This could have been caused by the pelleting process increasing the proportion of fine feed particles with fast disappearance from the bags. It was concluded that small associative effects between the examined single feeds could be disregarded when formulating compound feeds for dairy cows, and that additivity of EDCP and EDST can be assumed in most cases.

The objective of this study was to determine the effects of protease origin and dosage on the prececal (pc) amino acid (AA) digestibility and the influence on composition of the microbial community in the small intestine. In addition, the effects of phytase supplementation were investigated. A total of 8 dietary treatments were included. The basal diet contained mainly corn and soybean meal. Three protease products were added to the basal diet, each at the level recommended by the supplier and at an 8-fold level. Phytase was supplemented in another dietary treatment. Each dietary treatment was allocated to 8 replicates of 15 birds each. The experimental diets were offered from day 15 to 21 for ad libitum consumption. The effect of protease supplementation on the pc AA digestibility depended on the protease product type and the amount supplemented. The pc AA digestibility was significantly increased by 1 protease product when supplemented at high level and when phytase was supplemented. In all the other treatments, protease supplementation had no significant influence or it decreased pc AA digestibility, when compared with the treatment with no enzymes added. In general, Firmicutes was the most abundant phylum among the ileal microbiota across all the treatments. Significant effects on microbiota composition were observed at the genus level for some but not all protease treatments and phytase supplementation. The genera Streptococcus, Lactobacillus, and uncultured Clostridiaceae were responsible for these differences. Furthermore, microbial networks established for each diet showed either high or low number of intergeneric interactions, but without a consistent enzyme effect. We conclude that enzyme supplementation effects were evident in the terminal small intestine microbiota composition, and to a lesser extent, in pc AA digestibility. However, the changes in microbiota composition and pc AA digestibility could not be correlated, indicating absence of a causal relationship.

The aim of the present study was to test whether different dietary corn sources and phytase supplementation affect the prececal phosphorus digestibility (pcdP) and appearance of inositol phosphates in the lower ileum of growing broiler chickens and turkeys. Two experiments were conducted, one with broiler chickens and one with turkeys. Four corn diets were provided; these were formulated to contain low P and calcium (Ca) contents and incorporated 43% of one of the four different corn sources. Diets were either unsupplemented or supplemented with 500 FTU of an Escherichia coli-derived phytase/kg feed. Experimental diets were fed ad libitum from day 20 post-hatch. At 28 d of age, digesta were sampled from the lower ileum of animals to determine pcdP and pc myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and to analyze the concentrations of lower inositol phosphate isomers. The pcdP of non-supplemented diets ranged from 51 to 60% and from 22 to 28% in broilers and turkeys, respectively. A negative correlation was observed between the InsP6 content of the corn source and the pcdP of diets in broilers only. Without phytase supplementation, pc InsP6 degradation ranged from 64 to 76% in broilers and from 6 to 15% in turkeys. Phytase increased the pcdP by around 15% in broilers (P < 0.001) and 9 to 17% in turkeys (P < 0.001). In turkeys, phytase efficacy was greatest when the diets contained corn with higher contents of ether extract and InsP6. An effect of corn source on the appearance of lower InsPs in the ileal digesta was found in broilers only. These results suggest that broilers possess a greater capacity for InsP6 degradation and hydrolysis of lower InsPs compared with turkeys. Furthermore, the results are influenced by the corn source used. Further research is needed to identify the factors responsible for the low level of phytate degradation in turkeys in order to improve the availability of InsP6-P and the efficacy of phytase.

The objective of this study was to compare the effects of graded inclusions of 2 phytase products and a mineral P source in broiler chickens using different response traits, including ileum microbiota composition. Eleven experimental diets were used. These were a low-P basal diet and diets supplemented with increasing levels of dicalcium phosphate (DCP), Natuphos E 5000 G (NE), or Natuphos 5000 G (N). The performance traits, prececal P digestibility, and tibia and foot ash results were subjected to regression analysis and slope ratios were used to compare the supplements based on the measured evaluation traits. In the microbiota analysis, total nucleic acids were extracted and the 16S rRNA gene was targeted for use in the amplicon sequencing process. Phylogenetic analysis was performed using Mothur, followed by a multivariate statistical analysis. The various response traits caused different estimates of relative efficacy. The mean results of all the response traits showed that a 1.75-fold increase in the activity of N was needed to achieve the same response as NE and the variability among the detected traits ranged from 1.59 (prececally digestible P intake) to 1.91 (amount of tibia ash). The mean slope ratio between DCP and NE was 311 and varied between 208 (ADG) and 349 (foot ash concentration). The mean slope ratio for phytase N with DCP was 552 and varied from 357 (ADG) to 640 (tibia ash concentration). The ileum microbiota composition was not different among the diets. A similar composition was driven in the abundance of Lactobacillus crispatus, Lactobacillus salivarius, and Lactobacillus gallinarum. The results suggest that different response traits cause markedly different estimates of relative phytase efficacy.

Background Diet acidification, dietary calcium (Ca) level, and phytase supplementation are known influences on the microbial community in the digestive tract and on phosphorus (P) utilization of broiler chickens. Effects of dietary factors and microbiota on P utilization may be linked because microorganisms produce enzymes that release P from phytate (InsP6), the main source of P in plant feedstuffs. This study aimed to detect linkages between microbiota and InsP6 degradation by acidifying diets (i.e., replacing Ca carbonate (CaCO3) by Ca formate or adding formic acid to CaCO3-containing diets), varying Ca levels, and supplementing phytase in a three-factorial design. We investigated i) the microbial community and pH in the digestive tract, ii) prececal (pc) P and Ca digestibility, and iii) InsP6 degradation. Results All factors under investigation influenced digesta pH and the microbiota composition. Predicted functionality and relative abundance of microorganisms indicated that diets influenced the potential contribution of the microbiota on InsP degradation. Values of InsP6 degradation and relative abundance of the strains Lactobacillus johnsonii and Lactobacillus reuteri were correlated. Phytase supplementation increased pc InsP6 disappearance, with differences between Ca levels, and influenced concentrations of lower inositol phosphate isomers in the digestive tract. Formic acid supplementation increased pc InsP6 degradation to myo-inositol. Replacing CaCO3 by Ca-formate and the high level of these Ca sources reduced pc InsP6 disappearance, except when the combination of CaCO3 + formic acid was used. Supplementing phytase to CaCO3 + formic acid led to the highest InsP6 disappearance (52%) in the crop and increased myo-inositol concentration in the ileum digesta. Supplementing phytase leveled the effect of high Ca content on pc InsP6 disappearance. Conclusions The results point towards a contribution of changing microbial community on InsP6 degradation in the crop and up to the terminal ileum. This is indicated by relationships between InsP6 degradation and relative abundance of phosphatase-producing strains. Functional predictions supported influences of microbiota on InsP6 degradation. The extent of such effects remains to be clarified. InsP6 degradation may also be influenced by variation of pH caused by dietary concentration and solubility of the Ca in the feed.

A large F2 cross with 920 Japanese quail was used to map QTL for phosphorus utilization, calcium utilization, feed per gain and body weight gain. In addition, four bone ash traits were included, because it is known that they are genetically correlated with the focal trait of phosphorus utilization. Trait recording was done at the juvenile stage of the birds. The individuals were genotyped genome-wide for about 4k SNPs and a linkage map constructed, which agreed well with the reference genome. QTL linkage mapping was performed using multimarker regression analysis in a line cross model. Single marker association mapping was done within the mapped QTL regions. The results revealed several genome-wide significant QTL. For the focal trait phosphorus utilization, a QTL on chromosome CJA3 could be detected by linkage mapping, which was substantiated by the results of the SNP association mapping. Four candidate genes were identified for this QTL, which should be investigated in future functional studies. Some overlap of QTL regions for different traits was detected, which is in agreement with the corresponding genetic correlations. It seems that all traits investigated are polygenic in nature with some significant QTL and probably many other small-effect QTL that were not detectable in this study.

The objective of this study was to investigate the effect of variation in wheat-derived phytase activity on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation, inositol phosphate (InsP) isomer concentration and phosphorus (P) digestibility in pigs fed wheat-based diets. Additional effects of a microbial phytase supplementation were also studied. Three wheat genotypes (W1-W3) with an analysed phytase activity between 2760 and 3700 FTU/kg were used to formulate four experimental diets that included soybean meal and rapeseed meal but did not contain a mineral P supplement. DietW1-DietW3 only differed in the included wheat genotypes (W1-W3) at an inclusion level of 400 g/kg. DietW3+ contained W3 and a commercial 6-phytase supplementation at 500 FTU/kg diet. Eight barrows with an initial body weight of 27 kg were fitted with a simple T-cannula at the distal ileum and assigned to the four dietary treatments in a completely randomised row column design. The experiment included four periods of 12 d each. The first 5 d of each period were for diet adaptation, followed by collection of faeces (4 d), ileal digesta (2 d), and blood (last day). In DietW1-DietW3, the mean precaecal (pc) InsP6 disappearance was 48% and the mean pc P digestibility was 37% without a significant effect of the wheat genotype. The InsP6 disappearance measured in the faeces was close to complete in all treatments, and faecal P digestibility was not significantly affected by the wheat genotype (36% overall). The addition of microbial phytase caused a significant increase in pc InsP6 degradation (to 79%) and pc and total tract P digestibility (to 53% and 52%, respectively). The concentration of InsP6 degradation products in ileal digesta was not significantly affected by the wheat genotype, except for that of Ins(1,2,3,4,6)P5 and myo-inositol, which were higher in DietW3 than in DietW1 and DietW2. The added microbial phytase significantly reduced the concentration of InsP5 isomers in the ileal digesta and increased the concentrations of lower InsP isomers and myo-inositol. There were no significant effects of the added microbial phytase on pc amino acid digestibility; however, the wheat genotype exerted significant effects on the pc digestibility of Cys, Gly and Val. It was concluded that an increase in the intrinsic phytase activity of wheat achieved by crossbreeding was not reflected in InsP6 degradation and P digestibility in pigs fed wheat-based diets.

The study was conducted to determine the effects of high levels of phytase on growth performance, nutrient digestibility, phytate breakdown, and expression of mucosal tight junction and nutrient transporter genes in weanling pigs. A total of 128 barrows were penned in groups of four and used in a randomized completely block design and assigned to four treatments for a 28-d study. A two-phase feeding was implemented (phase 1: day 1 to 14; phase 2: day 15 to 28). The diets differed in dietary calcium (Ca) and phosphorus (P) levels (positive control [PC]: 8.1 to 7.1 g/kg Ca and 6.5 to 6.8 g/kg P; negative control [NC]: 6.6 to 5.5 g/kg Ca and 5.6 to 5.3 g/kg P) from phase 1 to phase 2, respectively. NC diets were supplemented with phytase at 0 (NC), 1,500 (NC + 1,500), or 3,000 (NC + 3,000) phytase units (FTU)/kg. Blood was collected after fasting (day 27) or feeding (day 28) for the measurement of plasma inositol concentrations. On day 28, two pigs per pen were euthanized. Duodenal–jejunal and ileal digesta samples and feces were collected to determine inositol phosphates (InsP3-6) concentrations. Phytase supplementation increased the body weight on days 14 and 28 (P < 0.05). Average daily gain and feed efficiency compared with NC were increased by phytase with the majority of its effect in phase 1 (P < 0.05). The apparent ileal digestibility and apparent total tract digestibility of P were increased in piglets fed phytase-supplemented diets (P < 0.01) compared with NC piglets. Disappearance of InsP6 and total InsP3-6 up to the duodenum–jejunum, ileum, and in feces was increased by both phytase application rates (P < 0.01). Plasma concentrations of myo-inositol were higher (P < 0.001) in the phytase-supplemented diets than PC and NC diets, irrespective of whether pigs were fed or fasted. Expression of claudin 3 was higher in pigs fed both phytase-supplemented diets in the duodenum and jejunum compared with PC and NC. Mucin 2 expression was lower in the ileum of NC + 3,000 fed piglets compared with PC (P < 0.05), whereas expression of GLUT2 (solute carrier family 2-facilitated glucose transporter member 2) was increased (P < 0.05) by the NC + 3,000 treatment in all sections. In summary, high phytase supplementation increased the growth performance of nursery pigs. The increased expression of GLUT2 by phytase may indicate an upregulation of glucose absorption from the intestine by phytase.

We investigated how the chemical composition of broiler chicken and cecectomized laying hen excreta is affected by drying in a forced-air drying chamber at low temperatures. Excreta that was immediately frozen after voiding provided the reference values. The excreta were dried in drying chambers for 4 hr, 6 hr, and 12 hr at 23°C or 33°C in the broiler experiment and 19°C or 29°C in the cecectomized laying hen experiment. The total N and inositol phosphate concentrations in the excreta of broiler chickens and cecectomized laying hens were not influenced (p > .050), except for one inositol tetrakisphosphate isomer (p = .026) in broilers. Compared to fresh excreta, drying did not affect the ammonia concentrations in the cecectomized laying hen experiment (p > .050), but the ammonia concentration was lower when dried for 12 hr at 33°C in the broiler experiment (p = .002). Amino acid concentrations in cecectomized laying hen excreta decreased until 4 hr of drying and then increased at both drying temperatures (p

The objectives of this study were to investigate the digestibility of pumpkin seed cake (PSC) for the rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), and effects on performance and product quality traits of four different fish species when PSC partially replaced fishmeal in extruded diets. A digestibility trial was carried out to determine apparent digestibility coefficients (ADC) for crude protein (CP), ether extract (EE) and gross energy (GE) of PSC fed to rainbow trout. In subsequent growth trials, effects on performance and morphological traits and fillet colour values of four different fish species [rainbow trout; brook trout, Salvelinus fontinalis (Mitchill, 1814); African sharptooth catfish, Clarias gariepinus (Burchell, 1822); and wels catfish, Silurus glanis (Linnaeus, 1758)] were evaluated when 60% of fishmeal protein of a reference diet was replaced by PSC protein (based on digestible CP). Nutrient ADC of PSC were high (CP: 89%, EE: 88% and GE: 84%). No significant effects on growth and only minor effects on fillet colour were detected in the trials. However, replacing fishmeal with PSC at the chosen level affected morphological traits and feed conversion in all four species to different extents. Replacement effects of PSC should be tested at lower levels of inclusion before conclusions are drawn on its suitability in fish diets.

This study aimed to distinguish between the single and interactive effects of phosphorus (P), calcium (Ca), and phytase on products of phytate degradation, including the disappearance of myo-inositol (MI), P, Ca, and amino acids (AA) in different segments of the digestive tract in broiler chickens. Additionally, all dephosphorylation steps from myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) to MI were investigated in the digesta of the terminal ileum. Unsexed Ross 308 broiler chickens were allocated to 56 pens with 19 birds per pen, and assigned to one of 8 dietary treatments. The dietary treatments included diets without (P−, 4.1 g/kg DM) or with (P+, 6.9 g/kg DM) monosodium phosphate supplementation, without (Ca−, 6.2 g/kg DM) or with (Ca+, 10.3 g/kg DM) additional fine limestone supplementation, and without or with 1,500 FTU phytase/kg feed in a factorial design. Adding Ca or P had no effect on InsP6 disappearance in the crop when phytase was added. InsP6 disappearance up to the terminal ileum (P−Ca− 56%) was decreased in P+Ca− (40%), and even more so in P+Ca+ (21%), when no phytase was added. Adding phytase removed all effects of P and Ca (77 to 87%); however, P+Ca+ increased the concentrations of lower InsP esters and reduced free MI in the ileum, even in the presence of phytase. These results indicate that mineral supplements, especially P and Ca combined, reduce the efficacy of endogenous microbial or epithelial phosphatases. Supplementation with phytase increased, while supplementation with Ca decreased the concentration of MI in all segments of the digestive tract and in blood plasma, demonstrating the ability of broilers to fully degrade phytate and absorb released MI. While AA disappearance was not affected by P or Ca, or an interaction among P, Ca, and phytase, it increased with the addition of phytase by 2 to 6%. This demonstrates the potential of the phytase used to increase AA digestibility, likely independent of P and Ca supply.

The objective of this study was to investigate the effects of supplementation with free myo-inositol (MI) or graded levels of phytase on inositol phosphate (InsP) degradation, concentrations of MI in the digestive tract and blood, bone mineralization, and prececal digestibility of amino acids (AA). Ross 308 broiler hatchlings were allocated to 40 pens with 11 birds each and assigned to one of 5 treatments. The birds were fed a starter diet until d 11 and a grower diet from d 11 to d 22. All diets were based on wheat, soybean meal, and corn. Birds were fed a control diet, calculated to contain adequate levels of all nutrients without (C) or with MI supplementation (C+MI), or one of 3 experimental diets that differed in phytase level (modified E. coli- derived 6-phytase; Phy500, Phy1500, or Phy3000 FTU/kg), with P and Ca levels adapted to the recommendations of the phytase supplier for a phytase level of 500 FTU/kg. The gain:feed ratio (G:F) was increased by MI or phytase in the starter+grower phase by 0.02 g/g. Prececal P and Ca digestibility, P and Ca concentration in blood serum, and tibia ash weight did not differ among treatments (P > 0.05). MI supplementation led to the highest MI concentration in the crop, ileum, and blood plasma across treatments. Phytase supplementation increased MI concentrations in the crop and ileum digesta in a dose-dependent manner and in plasma without any dose effect (P > 0.05). Prececal digestibility of some AA was increased by phytase. These outcomes indicate that MI might have been a relevant cause for the increase in G:F. Therefore, it is likely that the release of MI after complete dephosphorylation of phytate is one of the beneficial effects of phytase, along with the release of P and improvement in digestibility of other nutrients. Simultaneously, MI seems to have no diminishing effects on InsP degradation.

The aim of this study was to evaluate the P digestibility of different wheat and triticale genotypes in growing broiler chickens. Additionally, the relationships between physical or chemical properties of genotypes and P digestibility were determined. A low P, low Ca basal diet based on cornstarch and soybean meal was supplemented with 20% or 40% of 8 different wheat or triticale genotypes at the expense of cornstarch. Experimental diets were fed to broilers between d 20 and 26 of age before digesta samples were collected from their lower ileum for determination of prececal P digestibility (pcdP). Triticale-based diets had an average pcdP of 54%. Neither the concentration of triticale nor the genotype itself affected the pcdP of diets. The pcdP of triticale genotypes calculated by linear regression analysis ranged from 53% to 78%. No correlations were found between physical or chemical properties (viscosity, phytase activity, total and phytate P contents) and the pcdP of triticale genotypes. In contrast, a significant (P < 0.05) effect of genotype and wheat concentration was observed in wheat-based diets. The average pcdP of diets decreased from 60% to 50% by increasing the wheat concentration from 20% to 40%. As no linear relationship was observed between P intake and the amount of pcdP in the diet, the pcdP (%) of wheat genotypes was calculated separately for both concentrations, and accounted for 38% to 67% and 20% to 38% with 20% and 40% wheat inclusion in diets, respectively. Single chemical and physical characteristics could not explain the observed differences in P digestibility. Our results revealed a high variation in the pcdP of different wheat and triticale genotypes that should be considered in diet formulation. However, further research is needed to identify factors that contribute to variation of P digestibility in both grains.

1. Herein, it was investigated whether different particle size distributions of feed ingredients achieved by grinding through a 2- or 3-mm grid would have an effect on precaecal (pc) amino acid (AA) digestibility. Maize and soybean meal were used as the test ingredients. 2. Maize and soybean meal was ground with grid sizes of 2 or 3 mm. Nine diets were prepared. The basal diet contained 500 g/kg of maize starch. The other experimental diets contained maize or soybean meal samples at concentrations of 250 and 500, and 150 and 300 g/kg, respectively, instead of maize starch. Each diet was tested using 6 replicate groups of 10 birds each. The regression approach was applied to calculate the pc AA digestibility of the test ingredients. 3. The reduction of the grid size from 3 to 2 mm reduced the average particle size of both maize and soybean meal, mainly by reducing the proportion of coarse particles. Reducing the grid size significantly (P 0.050) increased the pc digestibility of all AA in the soybean meal. In maize, reducing the grid size decreased the pc digestibility of all AA numerically, but not significantly (P 0.050). The mean numerical differences in pc AA digestibility between the grid sizes were 0.045 and 0.055 in maize and soybean meal, respectively. 4. Future studies investigating the pc AA digestibility should specify the particle size distribution and should investigate the test ingredients ground similarly for practical applications.

The objective of the present study was to determine whether there are differences in prececal amino acid digestibility between commonly used slow- and fast-growing broiler strains when the regression approach is applied. ISA J-275 and Ross 308 were selected as common representatives of slow- and fast-growing broiler strains, respectively. The experimental diets with soybean cake at levels of 0, 100, and 200 g/kg were offered for ad libitum consumption between 22 and 29 d post-hatch. Titanium dioxide was used as an indigestible marker. Each treatment was tested with six pens comprising 10 birds each. Digesta samples were collected on a pen basis from the distal two-thirds of the intestine section between Meckel’s diverticulum and 2 cm anterior to the ileocecal-colonic junction. The prececal amino acid digestibility of soybean cake was calculated by linear regression simultaneously for both strains. There was no significant interaction between broiler strain and inclusion level of soybean cake with respect to the prececal CP and amino acid digestibility of complete diets; there was a significant strain effect for 5 out of the 16 measured amino acids. The prececal CP and amino acid digestibility of soybean cake did not differ significantly between strains and was numerically almost identical. The results of the present study provide evidence of the transferability between broiler strains of prececal amino acid digestibility data, determined using the regression approach, thus improving the accuracy of diet formulation without drawbacks.

To optimize the use of corn grain in diets for laying hens, differences in amino acid (AA) digestibility and metabolizable energy among different corn samples should be considered in feed formulation. The present study investigated the variability of AA digestibility and AMEn concentration of 20 corn samples in cecectomized laying hens. Corn grains were characterized based on their physical properties (thousand seed weight, test weight, grain density, and extract viscoelasticity), chemical composition (proximate nutrients, AA, minerals, and inositol phosphates), gross energy concentration, and in vitro solubility of nitrogen to study any relationship with AA digestibility or AMEn. The animal study comprised 4 Latin squares (6 × 6) distributed between 2 subsequent runs. Cecectomized LSL-Classic hens were individually housed in metabolism cages and fed either a basal diet containing 500 g/kg cornstarch or one of 20 corn diets, each replacing the cornstarch with one corn batch, for 8 days. During the last 4 d, feed intake was recorded and excreta were collected quantitatively. A linear regression approach was used to calculate AA digestibility of the corn. The digestibility of all AA differed significantly between the 20 corn batches, including Lys (digestibility range 64 to 85%), Met (86 to 94%), Thr (72 to 89%), and Trp (21 to 88%). The AMEn of the corn batches ranged between 15.7 and 17.1 MJ/kg DM. However, consistent correlations between AA digestibility or AMEn and the physical and chemical characteristics of the grains were not detected. Equations to predict AA digestibility or AMEn based on the grain's physical and chemical characteristics were calculated by multiple linear regressions. The explanatory power (adjusted R2;) of prediction equations was below 0.6 for the majority of AA and AMEn, and, thus, was not sufficiently precise for practical use. Possible explanations for the variation in AA digestibility and AMEn beyond the determined characteristics are discussed. In conclusion, AA digestibility and AMEn of corn grain is high in laying hens, but varies among different corn samples, with physical and chemical characteristics not suitable for explaining these variations.

Two experiments (Exp.) with ileally cannulated growing barrows were conducted. The concentrations of positional inositol phosphate (InsP) isomers in ileal digesta and feces were determined, as well as the prececal and total tract phytate (InsP6) hydrolysis, and digestibility of dry matter, P, Ca, nitrogen, and gross energy. Prececal amino acid (AA) digestibility and digestive enzyme activities in ileal digesta were also studied. In both Exp., pigs had an initial body weight (BW) of 28 kg and were completely randomized to a Double Latin Square Design with eight pigs, four diets, and three periods of 12 d each. Feces and ileal digesta were collected for 5 d and 2 d, respectively. Pigs were housed individually in stainless steel metabolic units. Water was available ad libitum and feed was provided two times daily at an amount of 4% of mean BW. In Exp. 1, pigs received a corn–soybean meal (SBM)-based diet that was supplemented with 0, 750, 1,500, or 3,000 FTU of a microbial phytase/kg diet. In Exp. 2, pigs were allotted to a 2 × 2 arrangement of diets based on corn and SBM or an SBM-rapeseed cake (RSC) mix and phytase supplementation at 0 or 1,500 FTU/kg of diet. In ileal digesta of pigs fed without the phytase supplement, the dominating InsP isomers beside InsP6 were InsP5 isomers. The InsP pattern in ileal digesta changed with the inclusion of microbial phytase in both Exp., as there was a remarkable increase in Ins(1,2,5,6)P4 concentration (P < 0.001). In both Exp., the myo-inositol concentration in ileal digesta was greater upon phytase addition (P < 0.001). Without phytase supplementation, prececal and total tract P digestibility were low, whereas hardly any InsP6 was excreted in feces. There was no difference between prececal and total tract P digestibility values. For most AA studied in Exp. 2, prececal digestibility was lower (P < 0.01) when the diet contained RSC. However, phytase supplementation did not significantly affect prececal AA digestibility in both Exp. The present study showed that InsP6 disappearance by the end of the ileum can be increased up to around 90% in SBM- and SBM–RSC-based diets when microbial phytase is supplemented, but prececal P digestibility hardly exceeded 60%. The study confirms that pigs cannot benefit from a remarkable InsP6 degradation in the hindgut.

This study was conducted to determine effects of high phytase use on growth performance, amino acid (AA) digestibility, intestinal phytate breakdown, and nutrient transporter expression in starter broiler chickens. Male Ross 308 chicks were allocated to 24 pens, at 15 birds/pen and assigned to one of 4 dietary treatments. Treatments were: a control diet (PCa+) that contained adequate levels of calcium (Ca) and phosphorus (P) for growing broiler chicks; a reduced Ca and P diet (PCa-:-1.5 g P/kg and -1.6 g Ca/kg), and 2 additional diets in which phytase was supplemented in the PCa- diet at 1,500 (PCa-Phy1500) and 3,000 (PCa-Phy3000) FTU/kg feed. A common starter diet was fed from day 1 to 8. From day 8 to 22, birds were fed the 4 experimental diets. On day 22, birds were killed for sample collection. From day 8 to 15, average daily gain and average daily feed intake were not different across treatments (P < 0.05) but gain-to-feed ratio (G:F) was reduced (P < 0.006) in the PCa- treatment compared with other treatments. There were no further performance differences, but a tendency of phytase treatments improving the overall G:F (P = 0.079; day 8-22). Up to both the duodenum-jejunum and ileum, phytate, P, and Ca disappearance were increased (P < 0.05) in the PCa-Phy1500 and PCa-Phy3000 treatments compared with PCa- treatment. Phytase dose dependently increased myoinositol (MI) concentration in the digesta from both the duodenum-jejunum and ileum (P < 0.001). The highest concentration of MI was found in the PCa-Phy3000 treatment. Plasma MI concentration was increased by phytase supplementation (P < 0.001). Prececal disappearance of Cys was lower (P < 0.05) in the PCa- treatment than in PCa1and PCa-Phy3000 treatment. Expression of MUC2 in the duodenum-jejunum was higher (P < 0.05) in the PCa-Phy3000 treatment than in other treatments. Phytase-induced hydrolysis of phytate led to elevated digesta and plasma MI concentrations and reduced digesta concentrations of phytate breakdown intermediates.

Background There is good evidence for a substantial endogenous phytase activity originating from the epithelial tissue or the microbiota resident in the digestive tract of broiler chickens. However, ionophore coccidiostats, which are frequently used as feed additives in broiler diets to prevent coccidiosis, might affect the bacterial composition and the abundance of phytase producers in the gastrointestinal tract. The aim of the present study was to investigate whether supplementation of a frequently used mixture of the coccidiostats Narasin and Nicarbazin alone or together with a phytase affects microbiota composition of the digestive tract of broiler chickens, characteristics of phytate breakdown in crop and terminal ileum, and precaecal phosphorus and crude protein digestibility. Results Large differences in the microbial composition and diversity were detected between the treatments with and without coccidiostat supplementation. Disappearance of myo-inositol 1,2,3,4,5,6-hexakis(dihydrogen phosphate) (InsP6) in the digestive tract, precaecal P digestibility, inorganic P in blood serum, and the concentration of inositol phosphate isomers in the crop and ileum digesta were significantly affected by phytase supplementation, but not by coccidiostat supplementation. Crude protein digestibility was increased by coccidiostat supplementation when more phosphate was available. Neither microbial abundance and diversity nor any other trait measured at the end of the experiment was affected by coccidiostat when it was only supplemented from day 1 to 10 of age. Conclusions The coccidiostats used herein had large effects on overall microbiota composition of the digestive tract. The coccidiostats did not seem to affect endogenous or exogenous phytase activity up to the terminal ileum of broiler chickens. The effects of phytase on growth, phosphorus digestibility, and myo-inositol release were not altered by the presence of the coccidiostats. The effects of phytase and coccidiostats on nutrient digestibility can be of significant relevance for phosphorus and protein-reduced feeding concepts if confirmed in further experiments. Electronic supplementary material The online version of this article (10.1186/s42523-019-0006-2) contains supplementary material, which is available to authorized users.

1. Dietary glycine equivalents (Glyequi) for glycine and serine represent the first-limiting non-essential amino acid in poultry diets. Targeted adjustment of essential amino acids and Glyequi in d...

The objective of this ring test was to investigate the prececal phosphorus (P) digestibility of soybean meal (SBM) in broiler chickens using the trial protocol proposed by the World's Poultry Science Association. It was hypothesized that prececal P digestibility of SBM determined in the collaborating stations is similar. Three diets with different inclusion levels of SBM were mixed in a feed mill specialized in experimental diets and transported to 17 collaborating stations. Broiler chicks were raised on commercial starter diets according to station-specific management routine. Then they were fed the experimental diets for a minimum of 5 d before content of the posterior half of the ileum was collected. A minimum of 6 experimental replicates per diet was used in each station. All diets and digesta samples were analyzed in the same laboratory. Diet, station, and their interaction significantly affected (P < 0.05) the prececal digestibility values of P and calcium of the diets. The prececal P digestibility of SBM was determined by linear regression and varied among stations from 19 to 51%, with significant differences among stations. In a subset of 4 stations, the prececal disappearance of myo- inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate)-P; InsP6-P) also was studied. The prececal InsP6-P disappearance correlated well with the prececal P digestibility. We hypothesized that factors influencing InsP6 hydrolysis were main contributors to the variation in prececal P digestibility among stations. These factors were probably related to the feeding and housing conditions (floor pens or cages) of the birds in the pre-experimental phase. Therefore, we suggest that the World's Poultry Science Association protocol for the determination of digestible P be should extended to the standardization of the pre-experimental period. We also suggest that comparisons of P digestibility measurements among studies are made only with great caution until the protocol is more refined. info:eu-repo/semantics/publishedVersion

Phosphorus utilization (PU) has received considerable attention in poultry nutrition. However, reliable estimates of genetic parameters for PU and related traits have largely not been reported until now; however, these are needed to assess whether selection for an improved PU would result in selection response. A large Japanese quail F2 cross was generated and 888 F2 individuals were phenotyped for PU, bodyweight gain (BWG), and feed per gain (F:G). Because it can reasonably be assumed that the interrelationships between these traits are complex, structural equation models were used. The structural coefficient λij describes the rate of change of trait I with respect to trait j for a model with a recursive effect of trait j on trait i Three recursive structural coefficients (λF:G,PU,λBWG,PU,λBWG,F:G) were selected a priori based on biological knowledge. The model was fitted using ASReml software. Standard errors of estimated variance components and genetic parameters were approximated using the delta method. The heritability of PU, F:G, and BWG were 0.136, 0.118, and 0.092. The structural coefficient[Formula: see text]indicates that an increase in PU leads to reduced and thus improved F:G. The estimate[Formula: see text]indicates that improved F:G leads to an increase in BWG. The overall effect of PU on BWG was[Formula: see text]i.e. an increase in PU of 1% leads to an increase of BWG of 0.374 g in the data collection period, which spanned five days. The phenotypic and genetic correlations were negative between PU and F:G as well as between BWG and F:G and were positive between PU and BWG. These correlations are driven by direct genetic effects (pleiotropic genes or genes being in linkage disequilibrium) as well as by indirect genetic effects (genes affecting trait j affected indirectly trait i). The application of structural equation models contributed to our understanding of the complex biological relationship between PU, F:G, and BWG in quails. PU shows a heritability that is sufficient to achieve a selection response when breeding for this very-hard-to-measure trait.

The objectives of this meta-analysis were to determine to which extent phosphorus (P) digestibility and digestible P concentration in pig diets were increased by phytase supplementation and to quantify factors that potentially influence effects of phytase supplementation. A data set with a total of 547 data lines was compiled from 88 experiments published in 74 peer-reviewed papers between 2007 and April 2019. An exponential model was determined as more suitable to describe the response of P digestibility to phytase supplementation than a polynomial model. Phytase supplementation increased P digestibility by 25.6 percentage points (standard error (SE) = 1.54) to a plateau at 64.9% (SE = 1.82). The digestible P concentration was increased by phytase supplementation in the order of 1.01 g/kg (SE = 0.102) to a plateau at 2.62 g/kg (SE = 0.122). Goodness-of-fit criteria were R2 = 0.780 and root mean square error = 7.55% for P digestibility, and R2 = 0.691 and root mean square error = 0.48 g/kg for digestible P concentration. Consideration of further factors such as mineral P supplementation (yes or no), ad libitum vs. restrictive feeding, mixed diets vs. single feed ingredients, sex and age of pigs did not increase the accuracy of prediction in this data set. Some of these traits exhibited responses, but they likely are artefacts generated through the imbalanced structure of the data set. Effects of dietary total P, phytate (InsP6), InsP6-P to total P ratio, and Ca on the effect of supplemented phytase were not quantifiable. The present meta-analysis showed that responses to phytase supplementation can be well predicted although variation in P digestibility and digestible P concentration in the data set was high. Overall, predicted effects of phytase on P digestibility well corresponded to predictions made 25 years ago.

Phosphorus is an essential mineral for all living organisms and a limited resource worldwide. Variation and heritability of phosphorus utilization (PU) traits were observed, indicating the general possibility of improvement. Molecular mechanisms of PU, including host and microbial effects, are still poorly understood. The most promising molecules that interact between the microbiome and host are microRNAs. Japanese quail representing extremes for PU were selected from an F2 population for miRNA profiling of the ileal tissue and subsequent association with mRNA and microbial data of the same animals. Sixty-nine differentially expressed miRNAs were found, including 21 novel and 48 known miRNAs. Combining miRNAs and mRNAs based on correlated expression and target prediction revealed enrichment of transcripts in functional pathways involved in phosphate or bone metabolism such as RAN, estrogen receptor and Wnt signaling, and immune pathways. Out of 55 genera of microbiota, seven were found to be differentially abundant between PU groups. The study reveals molecular interactions occurring in the gut of quail which represent extremes for PU including miRNA-16-5p, miR-142b-5p, miR-148a-3p, CTDSP1, SMAD3, IGSF10, Bacteroides, and Alistipes as key indicators due to their trait-dependent differential expression and occurrence as hub-members of the network of molecular drivers of PU.

Phosphorus (P) is an essential component for all living beings. Low P diets prompt phenotypic and molecular adaptations to maintain P homeostasis and increase P utilization (PU). Knowledge of the molecular mechanisms of PU is needed to enable targeted approaches to improve PU efficiency and thus lower P excretion in animal husbandry. In a previous population study, Japanese quail were subjected to a low P diet lacking mineral P and exogenous phytase. Individual PU was determined based on total P intake and excretion. A subset of 20 extreme siblings discordant for PU was selected to retrieve gene expression patterns of ileum (n = 10 per PU group). Sequencing reads have been successfully mapped to the current Coturnix japonica reference genome with an average mapping rate of 86%. In total, 640 genes were found to be differentially abundant between the low and high PU groups (false discovery rate &le, 0.05). Transcriptional patterns suggest a link between improved PU and mitochondrial energy metabolism, accelerated cell proliferation of enterocytes, and gut integrity. In assessing indicators of the efficient use of macro- and micronutrients, further research on turnover and proliferation rates of intestinal cells could provide an approach to improve P efficiency in poultry species.

Gnotobiotic broiler chickens were used to study interactive effects of supplemented phosphorus, calcium (PCa), and phytase (Phy) on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and release of myo-inositol in the digestive tract. In 2 subsequent runs, the chickens were subjected to 1 of 4 dietary treatments with and without PCa and Phy supplementation. Sanitized eggs were hatched in 8 germfree isolators, and a minimum of 9 male Ross 308 chickens were placed in each pen (total 16 pens). Treatments implemented on day 10 included gamma-irradiated diets without (PCa−; 4.1g P and 6.2g Ca/kg DM) or with (PCa+; 6.9g P and 10.4g Ca/kg DM) monosodium phosphate and limestone supplementation and without (Phy−) or with (Phy+) 1,500 FTU Phy/kg feed in a factorial arrangement. On day 15, digesta was collected from different sections of the intestinal tract and analyzed for InsP isomers and myo-inositol. The isolators did not remain germfree, but analysis of contaminants and results of InsP degradation indicated no or minor effects of the identified contaminants. Prececal InsP6 disappearance was 42% with the PCa− Phy− treatment and 17% with PCa+Phy−. No InsP3–4 isomers were found in the digesta of the terminal ileum in PCa− Phy−. The concentration of myo-inositol in the ileal digesta from PCa− Phy− (6.1μmol/g DM) was significantly higher than that from PCa+Phy− (1.7μmol/g DM), suggesting rapid degradation of the lower InsP isomers by mucosal phosphatases and their inhibition by PCa. Phytase supplementation increased InsP6 disappearance and prevented inhibitory effects of PCa supplements (72% in PCa− Phy+ and 67% in PCa+Phy+). However, PCa supplementation reduced the degradation of lower InsP isomers mainly in the posterior intestinal sections in the presence of Phy, resulting in significantly lower myo-inositol concentrations. It is concluded that mucosa-derived phosphatases might significantly contribute to InsP6 degradation in broiler chickens. The potential of mucosa-derived phosphatases to degrade InsP6 and lower InsP is markedly reduced by dietary PCa supplementation.

A regression approach was applied to determine the influence of feed provisioning prior to digesta sampling on precaecal (pc) amino acid (AA) digestibility in broiler chickens. Soybean meal was used as an example test ingredient. Five feed-provisioning protocols were investigated, four with restricted provision and one with ad libitum provision. When provision was restricted, feed was provided for 30 min after a withdrawal period of 12 h. Digesta were sampled 1, 2, 4 and 6 h after feeding commenced. A diet containing 300 g maize starch/kg was prepared. Half or all the maize starch was replaced with soybean meal in two other diets. Average pc digestibility of all determined AA in the soybean meal was 86% for the 4 and 6-h protocols and 66% and 60% for the 2 and 1-h protocols, respectively. Average pc AA digestibility of soybean meal was 76% for ad libitum feed provision. Feed provisioning also influenced the determined variance. Variance in digestibility ranked in magnitude 1 h ad libitum 2 h 6 h 4 h for all AA. Owing to the considerable influence of feed-provisioning protocols found in this study, comparisons of pc AA digestibility between studies applying different protocols prior to digesta sampling must be treated with caution. Digestibility experiments aimed at providing estimates for practical feed formulation should use feed-provisioning procedures similar to those used in practice.

The existing literature is inconsistent with respect to optimal dietary concentrations of glycine (Gly) and serine (Ser) in broiler feed. Therefore, we conducted a meta-analysis to investigate the response of broilers to dietary levels of Gly using a full quadratic model based on mixed model methodology. Response was measured as ADG (g/d), ADFI (g/d), and G:F (g/g). In addition, the influence of other dietary constituents was evaluated. This meta-analysis was based on a data set comprising a total of 9,626 broilers in 10 peer-reviewed papers that investigated the response of broilers to different dietary concentrations of Gly, achieved by addition of free Gly. The fitted quadratic model, with either Gly+Ser or the calculated glycine equivalent (Glyequi) of both amino acids as the independent variable, revealed that all model terms were significant (P ≤ 0.05), and hence proved a curvilinear relationship between these independent variables and response traits. The R(2) value and root MS error confirmed a strong relationship between observed and predicted traits. A comparison of the influence of Gly+Ser and Glyequi on response traits revealed that both approaches produced similar results. Because Glyequi should meet the physiological values of a diet better than Gly+Ser, models with 2 independent variables were conducted using Glyequi. The second independent variables were methionine (Met):TSAA ratio and the concentrations of cysteine (Cys) and CP. In models with one or 2 independent variables, the impact of dietary Gly on ADFI was low. By contrast, G:F was markedly influenced by dietary Gly; this effect intensified at lower Met:TSAA ratios and higher Cys and CP levels. ADG was also a function of Glyequi and the second independent variables. For ADG, an optimal Met:TSAA ratio of 0.655 and Cys concentration of 0.302% was calculated. Following the nonlinear nature of relationship, generally applicable replacement values could not be calculated. However, it was concluded that consideration of dietary Cys can diminish the requirement for Glyequi, and therefore, enable a reduction in the CP of broiler diets without limiting growth performance.

Differences in the optimal dietary concentrations of Gly (glycine) and Ser (serine) in broiler diets may be due to levels of endogenous Gly precursors that differ in literature. Therefore, we measured the extent of the interactive effects between equivalents of Gly and Ser (Glyequi) and the endogenous Gly precursors choline and Thr (threonine) on growth performance. A fractional central composite design included concentrations of 15-25 g/kg DM, 0.6-2.0 g/kg DM, and 6.4-10.4 g/kg DM for Glyequi, choline, and Thr, respectively, in 5 levels each. The various concentrations were achieved by adding Gly, choline chloride, and l-Thr to a basal mix. Except for 20 replicates of the central diet, all treatments were tested with 5 replicates, each with 10 birds. Food was provided for ad libitum consumption throughout the experiment. The data were evaluated using artificial neural networks. Digestibility was studied for selected diets using separate birds. Since average daily feed intake (ADFI) varied between replicates, the intake of prececal digestible Glyequi, choline, and prececal digestible Thr were more adequate independent variables than the dietary concentration of each amino acid. From d 1 to d 7, no treatment effects on G:F and average daily gain (ADG) were detected; subsequent results refer to the period from d 7 to d 21. Increasing prececal digestible Thr intake considerably decreased the need for prececal digestible Glyequi to achieve certain levels of G:F and ADG. The extent of this effect cannot be explained only by the endogenous metabolism of Thr to Gly. Since essential amino acids were present above the recommended levels, Thr probably limited performance, and excessive intake of other essential amino acids prompted a Gly-dissipating process. Choline exerted a considerable effect on the required intake of prececal digestible Glyequi and prececal digestible Thr to achieve certain levels of G:F and ADG. The results of this study partly explain the previously reported variations in response to dietary Thr, Gly, and Ser.

Phytate breakdown in the digestive tract of broilers is affected by supplements of mineral phosphorus (P) and phytase with unknown interactions between the 2 factors. It was the objective to study phytate hydrolysis and the presence of inositol phosphate isomers (InsPs) as affected by supplements of mineral P and phytase in the small intestine of broilers. Fifteen-day old broilers were assigned to 48 pens of 20 broilers each (n = 8 pens/treatment). Two low-P corn–soybean meal-based diets without (BD−; 4.4 g P/kg dry matter) or with monocalcium phosphate (MCP; BD+; 5.2 g P/kg dry matter) were supplied without or with added phytase at 500 or 12,500 FTU/kg. On d 24, digesta from the duodenum/jejunum and lower ileum was pooled per segment on a by-pen basis, freeze-dried, and analyzed for P, InsPs, and the marker TiO2. Another 180 broilers (n = 6 pens/treatment, 10 birds each) were fed the 3 BD+ diets from d 1 to 21 to assess the influence of supplemented phytase on tibia mineralization and strength. Significant interactions between MCP and phytase supplements on myo- inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) hydrolysis (duodenum/jejunum: P ≤ 0.001; ileum: P = 0.004) and level of specific lower InsPs were detected. Supplementation with 12,500 FTU/kg phytase resulted in 92% InsP6 hydrolysis and strong degradation of InsP5. This treatment resulted in higher P net absorption, affirmed by higher BW gain, tibia strength, and mineralization compared to treatments without or with 500 FTU/kg phytase (P ≤ 0.05). MCP supplementation reduced the degradation of InsP6 and specific lower InsPs in birds fed diets without or with 500 FTU/kg of phytase (P ≤ 0.05), but did not reduce InsP6 hydrolysis or degradation of InsP5 at the high phytase dose. Effects of added MCP on phytase efficacy depend on the dose of supplemented phytase. Differences in the concentrations of lower InsPs indicated that the initial step of InsP6 hydrolysis is not the only catabolic step that is influenced by MCP or phytase levels.