Microbial signatures and enterotype clusters in fattening pigs: implications for nitrogen utilization efficiency.

As global demand for pork continues to rise, strategies to enhance nitrogen utilization efficiency (NUE) in pig farming have become vital for environmental sustainability. This study explored the relationship between the fecal microbiota, their metabolites, and NUE in crossbreed fattening pigs with a defined family structure. Pigs were kept under standardized conditions and fed in a two-phase feeding regime. In each phase, one fecal sample was collected from each pig. DNA was extracted from a total of 892 fecal samples and subjected to target amplicon sequencing. The results indicated an influence of sire, sampling period (SP), and sex on the fecal microbiota. Streptococcus emerged as a potential biomarker in comparing high and low NUE pigs in SP 1, suggesting a genetic predisposition to NUE regarding the fecal microbiota. All fecal samples were grouped into two enterotype-like clusters named cluster LACTO and cluster CSST. Pigs' affiliation with enterotype-like clusters altered over time and might be sex-dependent. The stable cluster CSST demonstrated the highest NUE despite containing pigs with lower performance characteristics such as average daily gain, dry matter intake, and daily nitrogen retention. This research contributes with valuable insights into the microbiome's role in NUE, paving the way for future strategies to enhance sustainable pig production.

Genomic analyses of nitrogen utilization efficiency, its indicator trait blood urea nitrogen and the relationship to classical growth performance and feed efficiency traits in a Landrace × Piétrain crossbred population.

Improving the nutrient efficiency in pork production is required to reduce the resource competition between human food and animal feed regarding diet components edible for humans and to minimize emissions relevant to climate or the environment. Thereby, protein utilization efficiency and its equivalent nitrogen utilization efficiency (NUE) play a major role. Breeding for more nitrogen (N) efficient pigs bears a promising strategy to improve such traits, however, directly phenotyping NUE based on N balance data is neither cost-efficient nor straightforward and not applicable for routine evaluations. Blood urea nitrogen (BUN) levels in the pig are suitable to predict the NUE and, therefore, might be an indicator trait for NUE because BUN is a relatively easy-to-measure trait. This study investigated the suitability of NUE as a selection trait in future breeding programs. The relationships to classical growth performance and feed efficiency traits were analysed as well as the relationship to BUN to infer the role of BUN as an indicator trait to improve NUE via breeding. The analyzes were based on a Landrace F1 cross population consisting of 502 individuals who descended from 20 Piétrain sires. All animals were genotyped for 48,525 SNPs. They were phenotyped in two different fattening phases, i.e., FP1 and FP2, during the experiment. Uni- and bivariate analyses were run to estimate variance components and to determine the genetic correlation between different traits or between the same trait measured at different time points. Moderate heritabilities were estimated for all traits, whereby the heritability for NUE was h2 = 0.293 in FP1 and h2 = 0.163 in FP2 and BUN had the by far highest heritability (h2 = 0.415 in FP1 and h2 = 0.460 in FP2). The significant genetic correlation between NUE and BUN showed the potential of BUN to be considered an indicator trait for NUE. This was particularly pronounced when NUE was measured in FP1 (genetic correlations r g = - 0.631 $$ {r}_g=-0.631 $$ and r g = - 0.688 $$ {r}_g=-0.688 $$ between NUE and BUN measured in FP1 and FP2, respectively). The genetic correlations of NUE and BUN with important production traits suggest selecting pigs with high growth rates and low BUN levels to breed more efficient pigs in future breeding programs.

Impact of Trace Mineral Source and Phytase Supplementation on Prececal Phytate Degradation and Mineral Digestibility, Bone Mineralization, and Tissue Gene Expression in Broiler Chickens.

The objective of this study was to determine how different sources of Zn, Mn, and Cu in the feed without and with phytase affect prececal myo-inositol hexakisphosphate (InsP6) breakdown to myo-inositol (MI), prececal P digestibility, bone mineralization, and expression of mineral transporters in the jejunum of broiler chickens. A total of 896 male broiler chicks (Cobb 500) were distributed to 7 diets with 8 replicate pens (16 birds per floor pen). Experimental diets were fed from day 0 to 28. Diets were without or with phytase supplementation (0 or 750 FTU/kg) and were supplemented with three different trace mineral sources (TMS: sulfates, oxides, or chelates) containing 100 mg/kg Zn, 100 mg/kg Mn, and 125 mg/kg Cu. Prececal InsP6 disappearance and P digestibility were affected by interaction (phytase × TMS: P ≤ 0.010). In diets without phytase supplementation, prececal InsP6 disappearance and P digestibility were greater (P ≤ 0.001) in birds fed chelated minerals than in birds fed sulfates or oxides. However, no differences were observed between TMS in diets with phytase supplementation. Ileal MI concentration was increased by exogenous phytase but differed depending on TMS (phytase × TMS: P ≤ 0.050). Tibia ash concentration as well as Zn and Mn concentration in tibia ash were increased by phytase supplementation (P < 0.010), but the Cu concentration in tibia ash was not (P > 0.050). Gene expression of the assayed mineral transporters in the jejunum was not affected by diet (P > 0.050), except for Zn transporter 5 (phytase × TMS: P = 0.024). In conclusion, the tested TMS had minor effects on endogenous phytate degradation in the digestive tract of broiler chickens. However, in phytase-supplemented diets, the choice of TMS was not relevant to phytate degradation under the conditions of this study.

The influence of substituting dietary peptide-bound with free amino acids on nitrogen metabolism and acid-base balance of broiler chickens depends on asparagine and glutamine supply.

Reducing dietary crude protein (CP) concentration while maintaining adequate amino acid (AA) supply by free AA inclusion can contribute to attenuate the negative environmental effects of animal farming. This study investigated upper limits of dietary free AA inclusions without undesirable effects including the dependence on asparagine (Asn) and glutamine (Gln) supply. Ten broilers were allocated to sixty-three metabolism units each and offered nine experimental diets from day (d) 7-21 (n 7). One diet (167 g CP/kg) contained 80 g soya protein isolate (SPI)/kg. In the other diets, 25, 50, 75 and 100 % of the digestible AA from SPI were substituted with free AA. Digestible Asn+aspartic acid (Asp) and Gln+glutamic acid (Glu) were substituted with Asp/Glu or 50/50 mixes of Asp/Asn and Glu/Gln, respectively. Total excreta were collected from d 11-14 and from d 18-21. Growth and nitrogen accretion were unaffected by 25 and 50 % substitution without and with free Asn/Gln, respectively, but decreased at higher substitution (P ≤ 0·024). Circulating concentrations of Asp, Glu and Gln were unaffected by treatment, while Asn decreased at substitution higher than 50 % when Asn/Gln were not provided (P ≤ 0·005). Blood gas analysis on d 21 indicated a compensated metabolic acidosis at substitution higher than 50 and 75 % without and with free Asn/Gln, respectively (P ≤ 0·017). Results suggest that adding Asn/Gln increased an upper limit for proportion of dietary free AA from 10 to 19 % of dietary CP and enabled higher free AA inclusion without affecting the acid-base balance.

Effect of dietary zinc source, zinc concentration, and exogenous phytase on intestinal phytate degradation products, bone mineralization, and zinc status of broiler chickens.

This study aimed to determine the effect of Zn source and dietary level on intestinal myo-inositol hexakisphosphate (InsP6) disappearance, intestinal accumulation of lower InsP and myo-inositol (MI), prececal mineral digestibility, bone mineralization, and Zn status of broilers without and with exogenous phytase in the feed. Male Ross 308 broilers were allocated in groups of 10 to 8 treatments with 8 pens each. Experimental diets were fed from d 7 to d 28 and contained 33 mg/kg dry matter plant-intrinsic Zn. Experimental factors were phytase supplementation (0 or 750 FTU/kg) and Zn source (none [0 mg/kg Zn], Zn-sulfate [30 mg/kg Zn], Zn-oxide [30 mg/kg Zn]). Additional treatments with 90 mg/kg Zn as Zn-sulfate or Zn-oxide and phytase were included to test the effect of Zn level. No Zn source or Zn level effects were observed for ADG, feed conversion ratio, prececal P digestibility, intestinal InsP6 disappearance, and bone ash concentration. However, those measurements were increased by exogenous phytase (P < 0.001), except the feed conversion ratio, which was decreased (P < 0.001). Ileal MI concentrations were affected by phytase × Zn source interaction (P < 0.030). Birds receiving exogenous phytase and Zn supplementation had the highest MI concentrations regardless of exogenous Zn source, whereas MI concentrations were intermediate for birds receiving exogenous phytase only. Exogenous phytase and exogenous Zn source increased the Zn concentration in bone and blood of broilers (P < 0.001). In conclusion, measures of exogenous phytase efficacy were not affected by phytase × Zn source interaction. Further studies are needed to rule out an effect from Zn sources other than those tested in this study and to investigate the effect of Zn supplementation on endogenous phosphatases. The missing effect of increasing Zn supplementation from 30 to 90 mg/kg in phytase-supplemented diets gives reason to reconsider the Zn supplementation level used by the industry.

Nitrogen and lysine utilization efficiencies, protein turnover, and blood urea concentrations in crossbred grower pigs at marginal dietary lysine concentration.

Nitrogen utilization efficiency (NUE) and lysine utilization efficiency (LUE) are key indicators of sustainable pork production and vary depending on nutritional and non-nutritional factors. The objective was to study NUE and LUE together with concentrations of blood urea nitrogen (BUN) and other metabolites in growing pigs fed diets with marginal Lys concentrations at 11-13 wk (40.5 kg mean BW) and 14 to 16 wk (60.2 kg mean BW). The cereal grain-soybean meal-based diets contained 10.6 and 7.9 g Lys/kg DM in periods 1 and 2, respectively. Feed intake and BW were measured for 508 individually penned pigs, and blood samples were collected 5 h after morning feeding at weeks 13 and 16. A subgroup of 48 barrows was used in a nitrogen (N) metabolism trial at weeks 13 and 16. In this subgroup, the mean N retention of pigs (27.3 g N/d) and mean LUE (70%) were not different between the periods, but NUE was higher in period 1 (47%) than in period 2 (43%) (P < 0.001). After administration of a single dose of 15N labeled glycine and measurement of 15N recovery in urine, the calculated whole-body protein turnover did not differ between the periods. The rate of protein synthesis was positively correlated with NUE (P < 0.001), but protein degradation was not. Excretion of urea-N in urine accounted for 80% of the total urinary N and was positively correlated with BUN. The N retention of all 508 pigs was estimated using an equation that was derived from the N metabolism data. N retention was on average 31.4 g/d, equal in both periods, and higher in barrows than in gilts in period 2, but not in period 1 (P = 0.003). The calculated NUE was, on average, 47% and was lower in barrows than in gilts (P < 0.001) and higher in period 1 than in period 2 (P < 0.001). The calculated LUE was, on average, 71%, and was lower in barrows than in gilts in period 2, but not in period 1 (P < 0.001). The BUN concentration was higher in barrows than in gilts (P < 0.001) and higher in period 1 than in period 2 (P < 0.001). BUN concentration was negatively correlated with NUE in Periods 1 (r = -0.50) and 2 (r = -0.15) (P < 0.05). We concluded that the maximum LUE was in the range of 70-72% under the conditions of this study, and only small differences between the periods and sexes existed. Protein synthesis, rather than degradation, appears to affect NUE. BUN concentration may be useful for estimating NUE in a large group of animals fed a diet with a marginal Lys concentration.

Increasing the utilization of protein and amino acids by pigs is an effective tool for improving the sustainability of pork production. This study showed that the utilization of lysine, the limiting amino acid in pigs, can reach 72% in growing pigs aged 11­16 wk when fed diets based on cereal grains and soybean meal. The average utilization of total nitrogen was 47%, with high variation among the pigs. Higher nitrogen utilization was associated with lower concentration of urea in the blood. This indicates that the blood urea concentration is an easy-to-determine proxy trait for nitrogen utilization in pigs when the concentration of lysine in the feed is low. Only small differences were observed between gilts and barrows during the investigation period. Nitrogen utilization increased when pigs had a higher rate of whole-body protein synthesis, which was associated with anabolic hormone concentrations in the blood.

Effect of dietary calcium concentration and exogenous phytase on inositol phosphate degradation, mineral digestibility, and gut microbiota in growing pigs.

Variations in the dietary Ca concentration may affect inositol phosphate (InsP) degradation, and thereby, P digestibility in pigs. This study assessed the effects of dietary Ca concentration and exogenous phytase on InsP degradation, nutrient digestion and retention, blood metabolites, and microbiota composition in growing pigs with ileal cannulation. In a completely randomized row-column design with four periods, eight ileal-cannulated barrows (initial body weight 27 kg) were fed four corn-soybean- and rapeseed meal-based diets containing 5.5 or 8.5 g Ca/kg dry matter (DM), with or without 1,500 FTU of an exogenous hybrid-6-phytase/kg diet. No mineral P was added and the P concentration in the feed was 4.8 g P/kg DM. Prececal InsP6 disappearance in pigs fed diets containing exogenous phytase was lower (P = 0.022) with additional Ca than without. Concentrations of InsP2-4 isomers and myo-inositol in the distal ileal digesta and prececal P digestibility were greater (P < 0.001) with exogenous phytase than without exogenous phytase. In feces, InsP6 disappearance was lower (P < 0.002) and concentration of InsP5 and InsP4 isomers was higher (P ≤ 0.031) with additional Ca compared to without additional Ca. The prececal amino acid digestibility, energy digestibility, and hindgut disappearance of energy did not differ. The Shannon diversity index of the microbiota in the distal ileal digesta and feces was similar among the diets but was lower in the distal ileal digesta than in the feces (P < 0.001). Permutation analysis of variance revealed no dietary differences between the bacterial groups within the ileal digesta and fecal samples (P > 0.05). In conclusion, additional Ca reduced the effect of exogenous phytase on prececal InsP6 degradation. Endogenous InsP degradation was impaired by additional Ca only in the hindgut but the abundance of bacterial genera in feces was not affected.

The dietary calcium concentration can influence the release of phosphorus from phytate in growing pigs. This study assessed the effects of dietary calcium and exogenous phytase on inositol phosphate (InsP) degradation and nutrient digestibility in ileal-cannulated, growing pigs. The phosphorus, calcium, and myo-inositol concentrations in the blood, microbiota composition in the ileal digesta and feces, and volatile fatty acid concentrations in the feces were also evaluated. Additional dietary calcium decreased prececal inositol hexakisphosphate (InsP6) disappearance, but only with exogenous phytase. Concentrations of InsP2-4 isomers and myo-inositol in the ileal digesta and prececal phosphorus digestibility were greater with exogenous phytase, but not affected by dietary calcium concentration. In contrast, fecal InsP6 disappearance was lower and the concentration of InsP4-5 isomers in feces was greater with additional dietary calcium. Regarding microbiota, the Shannon diversity index was lower in ileal digesta than in feces but was unaffected by dietary calcium concentration or exogenous phytase. In conclusion, dietary calcium concentration is relevant for phytate disappearance in feces, but not in the ileal digesta. However, when exogenous phytase is used, the dietary calcium concentration is important because prececal phytate degradation is changed.

Interactions of zinc with phytate and phytase in the digestive tract of poultry and pigs: a review.

Phytase supplementation is gaining importance in animal nutrition because of its effect on phosphorus (P) digestibility and the increasing relevance of P for sustainable production. The potential inhibitors of phytase efficacy and phytate degradation, such as calcium (Ca) and zinc (Zn), have been a subject of intense research. This review focuses on the interactions of Zn with phytate and phytase in the digestive tract of poultry and pigs, with an emphasis on the effects of Zn supplementation on phytase efficacy and P digestibility. In vitro studies have shown the inhibitory effect of Zn on phytase efficacy. However, relevant in vivo studies are scarce and do not show consistent results for poultry and pigs. The results could be influenced by different factors, such as diet composition, amount of Zn supplement, mineral concentrations, and phytase supplementation, which limit the comparability of studies. The chosen response criteria to measure phytase efficacy, which is mainly tibia ash, could also influence the results. Compared to poultry, the literature findings are somewhat more conclusive in pigs, where pharmacological Zn doses (≥ 1000 mg kg-1 Zn) appear to reduce P digestibility. To appropriately evaluate the effects of non-pharmacological Zn doses, further studies are needed that provide comprehensive information on their experimental setup and include measurements of gastrointestinal phytate degradation to better understand the mechanisms associated with Zn and phytase supplements. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effect of increasing dietary fermentable fiber on diet nutrient digestibility and estimation of endogenous phosphorus losses in growing pigs.

Fermentable fiber may increase endogenous losses of phosphorus (EPL) and amino acids (AA), thereby reducing apparent nutrient digestibility. Acacia gum with medium-to-high fermentability and low viscosity was increasingly included in diets to investigate its effect on apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients, gross energy (GE), and standardized total tract digestibility (STTD) of P in growing pigs. A control diet (49% cornstarch; 18% bovine plasma protein) was formulated to measure basal EPL. Three additional diets were formulated to include 2.5%, 5.0%, or 7.5% acacia gum at the expense of cornstarch. Diets contained 16.1% to 17.4% CP and 0.31% to 0.33% total P (DM-basis). The four diets were fed to eight ileal-cannulated barrows (initial BW, 54.6 kg) for four 9 d periods in a double 4 × 4 Latin square. Apparent hindgut fermentation (AHF) was calculated as ATTD minus AID. Feeding increasing acacia gum quadratically affected (P < 0.05) AID of DM, GE, linearly decreased (P < 0.05) ATTD of DM, crude protein (CP), GE, digestible (DE) and predicted net energy (NE) value of diets, and linearly increased (P < 0.001) AHF of DM and GE. Increasing acacia gum did not affect AID and standardized ileal digestibility (SID) of CP and AA. Basal EPL was 377 mg/kg DM intake (DMI) and increasing acacia gum linearly increased (P < 0.05) total tract EPL. Increasing acacia gum linearly decreased (P < 0.05) diet ATTD of P, and STTD of P based on either the calculated EPL or NRC (2012) recommended value (190 mg P/kg DMI). Increasing acacia gum did not affect AID and ATTD of Ca of diets. In conclusion, feeding increasing dietary fermentable, low-viscous acacia gum decreased diet AID and ATTD of DM and GE, but did not affect AID or SID of CP and AA. Increasing acacia gum decreased ATTD of P, which might have been due to increased specific endogenous losses of P in the total tract of growing pigs.

Fermentable fiber may increase endogenous losses of phosphorus (EPL) and amino acids (AA), thereby reducing nutrient digestibility. This study assessed effects of feeding increasing acacia gum with medium-to-high fermentability and low viscosity. Eight barrows cannulated at the terminal ileum were fed a control diet to measure basal endogenous P losses and three diets including 2.5%, 5.0%, or 7.5% acacia gum. Increasing acacia gum quadratically decreased diet ileal digestibility of dry matter (DM), gross energy (GE), linearly decreased diet total tract digestibility of DM, crude protein (CP), GE, and linearly increased hindgut fermentation of DM and GE. Increasing acacia gum linearly increased total tract endogenous P losses. Increasing acacia gum linearly decreased diet total tract digestibility of P, and standardized total tract digestibility of P calculated based on the calculated endogenous P losses or table values (NRC, 2012; 190 mg P/kg DMI). In conclusion, increasing dietary fermentable, low-viscous acacia gum decreased diet ileal and total tract digestibility of DM and GE, but did not affect ileal digestibility of CP and most AA. Increasing acacia gum inclusion reduced total tract digestibility of P, which might have been due to increased specific endogenous losses of P in the total tract of growing pigs.

Mineral and Phytic Acid Content as Well as Phytase Activity in Flours and Breads Made from Different Wheat Species.

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.

Inferring causal structures of gut microbiota diversity and feed efficiency traits in poultry using Bayesian learning and genomic structural equation models.

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 (J'). 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 J', PU, and PR were quantified with structural equation models (SEM), which revealed a causal link from J' 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.

Feed efficiency and phosphorus efficiency are of increasing importance in poultry breeding. It was frequently shown that next to the birds' genomes also the gut microbiota composition is important for these efficiency traits. The gut microbiota composition is a mediator between the genomes of the birds and their efficiency traits. In the present study, an approach was taken to consider the animal's gut microbiota diversity, efficiency traits, and the genomes of the animals together in a causal network to decipher the mediator role between the traits. Growing Japanese quail were used as model species. A stable network could be established that placed the diversity of the gut microbiota composition at the forefront, with direct and indirect links to other traits like phosphorus utilization and retention, feed per gain ratio, and growth. Together with genome scans, the results confirmed the mediator role of the gut microbiota composition because several traits associated variants affected the efficiency traits directly and indirectly via the gut microbiota composition.

Feed Clusters According to In Situ and In Vitro Ruminal Crude Protein Degradation.

Effective degradation (ED) of crude protein (CP) was estimated in vitro at 0.02, 0.05 and 0.08 h−1 assumed ruminal passage rates for a total of 40 feedstuffs, for which in situ ED was available and used as reference degradation values. For this, the Streptomyces griseus protease test was used. The differences between in vitro CP degradation and the in situ CP degradation values were lowest in legume grains and highest in cereal by-products and barley. The differences between in situ and in vitro ED were expressed using a degradation quotient (degQ), where degQ = (EDin vitro − EDin situ)/EDin situ. Among the tested feedstuffs, eight specific clusters were identified according to degQ for the assumed passage rates. The feedstuffs clustered in an unspecific way, i.e., feedstuffs of different nutrient composition, origin or treatment did not necessarily group together. Formaldehyde−treated rapeseed meal, soybean meal, wheat, a treated lupin, sunflower meal and barley could not be assigned to any of the clusters. Groupwise degradation (range of degQ for assumed passage rates are given in brackets) was detected in grass silages (−0.17, −0.11), cereal by-products together with sugar beet pulp (−0.47, −0.35) and partly in legume grains (−0.14, 0.14). The clustering probably based on different specific nutrient composition and matrix effects that influence the solubility of feed protein and limit the performance of the protease. The matrix can be affected by treatment (chemically, thermally or mechanically), changing the chemical and physical structure of the protein within the plant. The S. griseus protease test had reliable sensitivity to reflect differences between native feedstuffs and treatments (thermally or chemically) that were found in situ. The in situ results, however, are mostly underestimated. The clustering results do not allow a clear conclusion on the groupwise or feed-specific use of carbohydrate-degrading enzymes as pre- or co-inoculants as part of the S. griseus protease test and need to be tested for its potential to make this test more conform with in situ data.

Comparison of mucosal phosphatase activity, phytate degradation, and nutrient digestibility in 3-week-old turkeys and broilers at different dietary levels of phosphorus and phytase.

A comparison between 3-wk-old female turkeys (B.U.T. 6) and broilers (Ross 308) was performed to study the effects of species, dietary P, Ca, and phytase levels on gut mucosal phosphatase activity, myo-inositol hexakisphosphate (InsP6) degradation along the digestive tract, digestibility of P, Ca, and amino acids, and concentrations of myo-inositol in the digesta and blood. The experimental diets were corn-soybean meal-based and identical for both species. Two dietary P and Ca concentrations (CaP-: 4.1 g P/kg, 5.5 g Ca/kg and CaP+: 9.0 g P/kg, 12.0 g Ca/kg) and 2 levels of phytase supplementation (0 and 1,500 FTU/kg) were used in a 2 × 2 factorial design and fed to the animals for 7 d in their third week of age. Each diet was randomly assigned to 6 broiler and 6 turkey pens, with 10 birds each. After slaughter, blood, digesta from the crop, gizzard, duodenum, lower ileum, and mucosa from the jejunum were collected. When fed CaP- without phytase supplementation, there were no differences between species in gut mucosal phosphatase activity, prececal InsP6 disappearance, and P and Ca digestibility, indicating a similar intrinsic capacity for phytate degradation in both species. When fed CaP+ without phytase supplementation, turkeys showed higher prececal InsP6 disappearance than broilers. Phytase supplementation increased prececal InsP6 disappearance and digestibility of P and Ca in both species. However, the phytase-induced increase in prececal InsP6 disappearance was more pronounced in broilers than in turkeys, possibly due to more adequate conditions for phytase activity in the broiler crop. In broilers, phytase supplementation increased amino acid digestibility overall, whereas, in turkeys, it increased with CaP+ and decreased with CaP-. In addition, the relationship between myo-inositol concentration in the ileum and blood differed between species, indicating differences in myo-inositol metabolism. It was concluded that 3-week-old turkeys and broilers differ in nutrient digestibility and InsP degradation in some segments of the digestive tract but have similar endogenous InsP6 degradation when fed low P and Ca diets.

Mucosal phosphatase activity, phytate degradation, and mineral digestibility in 6-week-old turkeys and broilers at different dietary levels of phosphorus and phytase and comparison with 3-week-old animals.

Female turkeys (B.U.T. 6) and broilers (Ross 308) were compared at 6 wk of age to evaluate the effects of species, dietary P, Ca, and phytase levels on myo-inositol hexakisphosphate (InsP6) degradation along the digestive tract, gut mucosal phosphatase activity, P and Ca digestibility, and myo-inositol concentrations in the digesta and blood. The environmental conditions and experimental corn-soybean meal-based diets were the same for both species. Four diets with either combination of 2 levels of P and Ca (CaP-: 4.0 g P/kg, 5.4 g Ca/kg and CaP+: 6.0 g P/kg, 8.0 g Ca/kg) and 2 levels of phytase supplementation (0 and 1,500 FTU/kg) were fed to the animals for 7 d at their sixth wk of age. Each diet was randomly assigned to 6 pens per species, with 10 birds each. After slaughter, blood, digesta from the crop, gizzard, duodenum, lower ileum, and jejunal mucosa were collected. Endogenous mucosal phosphatase activity in the jejunum was higher in turkeys than in broilers. Prececal InsP6 disappearance was also higher in turkeys than in broilers when phytase was not supplemented. Phytase supplementation led to a higher prececal InsP6 disappearance in broilers than in turkeys, likely due to different crop conditions such as moisture content. However, prececal P digestibility was higher in turkeys than broilers. Different relationships between myo-inositol concentration in the ileum digesta and blood were found, depending on the species. A comparison of the results with those obtained in 3-wk-old birds of a companion study showed that in diets with low Ca and P levels, prececal InsP6 disappearance increased with age in turkeys, but not in broilers. This coincided with changes in the conditions of the digestive tract, such as the water content in the crop, gizzard pH, and mucosal phosphatase activity. In conclusion, occurrence of differences in phytate degradation between turkeys and broilers, fed the same feed, depended on age and can be explained by different physiological development of the digestive tract.

The dynamics of molecular, immune and physiological features of the host and the gut microbiome, and their interactions before and after onset of laying in two hen strains.

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.

Minimum phosphorus requirements for laying hen feed formulations.

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.

The active core microbiota of two high-yielding laying hen breeds fed with different levels of calcium and phosphorus.

The nutrient availability and supplementation of dietary phosphorus (P) and calcium (Ca) in avian feed, especially in laying hens, plays a vital role in phytase degradation and mineral utilization during the laying phase. The required concentration of P and Ca peaks during the laying phase, and the direct interaction between Ca and P concentration shrinks the availability of both supplements in the feed. Our goal was to characterize the active microbiota of the entire gastrointestinal tract (GIT) (crop, gizzard, duodenum, ileum, caeca), including digesta- and mucosa-associated communities of two contrasting high-yielding breeds of laying hens (Lohmann Brown Classic, LB; Lohmann LSL-Classic, LSL) under different P and Ca supplementation levels. Statistical significances were observed for breed, GIT section, Ca, and the interaction of GIT section x breed, P x Ca, Ca x breed and P x Ca x breed (p < 0.05). A core microbiota of five species was detected in more than 97% of all samples. They were represented by an uncl. Lactobacillus (average relative abundance (av. abu.) 12.1%), Lactobacillus helveticus (av. abu. 10.8%), Megamonas funiformis (av. abu. 6.8%), Ligilactobacillus salivarius (av. abu. 4.5%), and an uncl. Fusicatenibacter (av. abu. 1.1%). Our findings indicated that Ca and P supplementation levels 20% below the recommendation have a minor effect on the microbiota compared to the strong impact of the bird's genetic background. Moreover, a core active microbiota across the GIT of two high-yielding laying hen breeds was revealed for the first time.

Methane Reduction Potential of Brown Seaweeds and Their Influence on Nutrient Degradation and Microbiota Composition in a Rumen Simulation Technique.

This study aimed to investigate the effects of two brown Icelandic seaweed samples (Ascophyllum nodosum and Fucus vesiculosus) on in vitro methane production, nutrient degradation, and microbiota composition. A total mixed ration (TMR) was incubated alone as control or together with each seaweed at two inclusion levels (2.5 and 5.0% on a dry matter basis) in a long-term rumen simulation technique (Rusitec) experiment. The incubation period lasted 14 days, with 7 days of adaptation and sampling. The methane concentration of total gas produced was decreased at the 5% inclusion level of A. nodosum and F. vesiculosus by 8.9 and 3.6%, respectively (P < 0.001). The total gas production was reduced by all seaweeds, with a greater reduction for the 5% seaweed inclusion level (P < 0.001). Feed nutrient degradation and the production of volatile fatty acids and ammonia in the effluent were also reduced, mostly with a bigger effect for the 5% inclusion level of both seaweeds, indicating a reduced overall fermentation (all P ≤ 0.001). Microbiota composition was analyzed by sequencing 16S rRNA amplicons from the rumen content of the donor cows, fermenter liquid and effluent at days 7 and 13, and feed residues at day 13. Relative abundances of the most abundant methanogens varied between the rumen fluid used for the start of incubation and the samples taken at day 7, as well as between days 7 and 13 in both fermenter liquid and effluent (P < 0.05). According to the differential abundance analysis with q2-ALDEx2, in effluent and fermenter liquid samples, archaeal and bacterial amplicon sequence variants were separated into two groups (P < 0.05). One was more abundant in samples taken from the treatment without seaweed supplementation, while the other one prevailed in seaweed supplemented treatments. This group also showed a dose-dependent response to seaweed inclusion, with a greater number of differentially abundant members between a 5% inclusion level and unsupplemented samples than between a 2.5% inclusion level and TMR. Although supplementation of both seaweeds at a 5% inclusion level decreased methane concentration in the total gas due to the high iodine content in the seaweeds tested, the application of practical feeding should be done with caution.

Multi-Omics Reveals Different Strategies in the Immune and Metabolic Systems of High-Yielding Strains of Laying Hens.

Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) are two commercially important laying hen strains due to their high egg production and excellent commercial suitability. The present study integrated multiple data sets along the genotype-phenotype map to better understand how the genetic background of the two strains influences their molecular pathways. In total, 71 individuals were analyzed (LB, n = 36; LSL, n = 35). Data sets include gut miRNA and mRNA transcriptome data, microbiota composition, immune cells, inositol phosphate metabolites, minerals, and hormones from different organs of the two hen strains. All complex data sets were pre-processed, normalized, and compatible with the mixOmics platform. The most discriminant features between two laying strains included 20 miRNAs, 20 mRNAs, 16 immune cells, 10 microbes, 11 phenotypic traits, and 16 metabolites. The expression of specific miRNAs and the abundance of immune cell types were related to the enrichment of immune pathways in the LSL strain. In contrast, more microbial taxa specific to the LB strain were identified, and the abundance of certain microbes strongly correlated with host gut transcripts enriched in immunological and metabolic pathways. Our findings indicate that both strains employ distinct inherent strategies to acquire and maintain their immune and metabolic systems under high-performance conditions. In addition, the study provides a new perspective on a view of the functional biodiversity that emerges during strain selection and contributes to the understanding of the role of host-gut interaction, including immune phenotype, microbiota, gut transcriptome, and metabolome.

The impact of dietary calcium content on phosphorus absorption and retention in growing pigs is enhanced by dietary microbial phytase supplementation.

Sixty growing male pigs were used to test the hypothesis that high dietary Ca content reduces P absorption to a greater extent in microbial phytase-supplemented diets via reducing inositol phosphate (IP) degradation and enhancing P precipitation. Pigs were equally allotted over diets with three Ca contents 2·0, 5·8 and 9·6 g/kg with or without microbial phytase (0 v. 500 FTU/kg) in a 2 × 3 factorial arrangement. Faeces and urine were collected at the end of the 21-d experimental period. Subsequently, pigs were euthanised and digesta quantitatively collected from different gastrointestinal tract (GIT) segments. Increasing dietary Ca content reduced apparent P digestibility in all GIT segments posterior to the stomach (P < 0·001), with greater effect in phytase-supplemented diets in the distal small intestine (Pinteraction = 0·007) and total tract (Pinteraction = 0·023). Nonetheless, increasing dietary Ca to 5·8 g/kg enhanced P retention, but only in phytase-supplemented diets. Ileal IP6 degradation increased with phytase (P < 0·001) but decreased with increasing dietary Ca content (P = 0·014). Proportion of IP esters in total IP (∑IP) indicated that IP6/∑IP was increased while IP4/∑IP and IP3/∑IP were reduced with increasing dietary Ca content and also with a greater impact in phytase-supplemented diets (Pinteraction = 0·025, 0·018 and 0·009, respectively). In all GIT segments, P solubility was increased with phytase (P < 0·001) and tended to be reduced with dietary Ca content (P < 0·096). Measurements in GIT segments showed that increasing dietary Ca content reduced apparent P digestibility via reducing IP degradation and enhancing P precipitation, with a greater impact in phytase-supplemented diets due to reduced IP degradation.