Effects of a Novel Consensus Bacterial 6-Phytase Variant on Growth Performance and Bone Ash of Broilers Fed Complex Diets Highly Deficient in Minerals, Digestible Amino Acids and Energy through 42 Days of Age.

An experiment was conducted to evaluate the effects of increasing the dose of a novel consensus bacterial 6-phytase variant expressed in Trichoderma reesei (PhyG) in broilers fed complex diets highly deficient in minerals, dig AA, and energy. Diets were a nutrient-adequate control (PC); a nutrient-reduced control (NC) formulated with a reduction in available P (avP) by 0.199%, Ca by 0.21%, crude protein by 0.72-1.03%, dig Lys by 0.064-0.084%, Na by 0.047%, and ME by 87.8 kcal/kg, respectively; and NC supplemented with PhyG at 500, 1000, and 2000 FTU/kg feed. BW was decreased and FCR increased in the NC vs. PC, while the PhyG treatments were similar to the PC. Carcass yield and bone ash were also maintained with PhyG supplementation. Phytase provided economic benefit on a feed cost per kg of weight basis for 1 to 35 d; the cost reductions equated to USD 0.006, 0.016, and 0.02/kg BWG at 500, 1000, and 2000 FTU/kg. In conclusion, this trial demonstrated that supplementation with a novel consensus phytase variant in diets highly deficient in minerals, dig AA, and energy maintained growth performance and provided economic benefit, with production benefits being maximized at inclusion levels of 2000 FTU/kg.

Application of enzyme matrix values for energy and nutrients to a wheat-corn-soybean meal-based broiler diet supplemented with a novel phytase, with or without a xylanase-ß-glucanase, achieved a production benefit over a nutritionally adequate unsupplemented diet.

The effect of applying an energy and nutrient matrix to a wheat-corn-soybean meal-based diet supplemented with a novel consensus bacterial 6-phytase variant (PhyG) and xylanase-ß-glucanase on growth performance, bone mineralization, carcass weights, feed costs, and carbon footprint was evaluated. A randomized complete block design (3,300 Ross 308 mixed-sex birds; 60 pens, 12 pens per treatment) tested 5 treatments: 1) a positive control diet (PC), containing 0.92, 0.84, 0.71% Ca and 0.43, 0.38, 0.30% digestible P during 1 to 10, 11 to 21, and 22 to 32 d of age, respectively; 2) a negative control reduced in Ca, digestible P, digestible AA, ME, and Na by phase based on the PhyG dosing regimen (NC1); 3) NC1 supplemented with PhyG at 2,000, 1,500, and 1,000 FTU/kg by phase (NC1+PhyG); 4) as NC1 but additionally reduced in ME (NC2); and 5) NC2 supplemented with PhyG as in 3) plus 1,220 U/kg of xylanase and 152 U/kg of ß-glucanase (NC2+PhyG+XB). Final (d 32) BW, overall (0-32 d of age) ADFI, FCR, d 10 and 32 tibia ash and carcass part weights were reduced or impaired (P < 0.05) in NC1 and NC2 vs. PC (d 32 BW -477 g/bird (23.4%) and -422 g/bird (20.7%), respectively). Growth performance (all measures, all phases) was improved and tibia ash (at 10 and 32 d of age), total carcass thigh, breast and leg weights were increased (P < 0.05) in NC1+PhyG vs. NC1, and NC2+PhyG+XB vs. NC2. Overall growth performance outcomes in NC1+PhyG and NC2+PhyG+XB were not different (P > 0.05) from the PC. Total feed cost and carbon footprint per kilogram BW gain (BWG) were reduced (P < 0.05) vs. PC in NC2+PhyG+XB [-0.052 € and -376 g CO2 eq./kg BWG, respectively] and NC1+PhyG [-0.038 € and -260 g CO2 eq./kg BWG, respectively]. The results validated the nutrient matrices in the test diets and highlighted a potential feed cost and environmental sustainability benefit which was greatest when the enzymes were applied in combination.

Effect of a biosynthetic bacterial 6-phytase on the digestibility of phosphorus and phytate in midlactating dairy cows.

The effect of a biosynthetic bacterial 6-phytase (PhyG) on the digestibility and excretion of crude protein (CP), phosphorus (P), and phytate-P (PP) in midlactating dairy cows was investigated. Thirty Holstein-Friesians were assigned to three treatments with 10 cows per treatment in a randomized block design. Cows were fed forage (grass and corn silage) provided ad libitum, and a concentrate (without added inorganic phosphate) administered separately in amounts individualized per cow according to milk production, supplemented with phytase according to treatment. The formulated forage-to-concentrate-ratio was ~65%:35%. Dietary treatments comprised the control diet (CON) and CON supplemented with 2,000 (PhyG2,000) or 5,000 (PhyG5,000) phytase units (FTU)/kg DM in the total diet. The experiment comprised an 18-d preperiod for the collection of data to facilitate the allocation of cows to the treatments, followed by a 19-d experimental period comprising a 14-d diet adaptation period and 5 d of twice daily feces collection. Fecal samples were analyzed for the determination of apparent total tract digestibility (ATTD) of chemical constituents in the diet. The ATTD of PP was 92.6% in CON suggesting a high but incomplete degradation of phytate by ruminal microbial phytases. Cows fed PhyG2,000 exhibited increased ATTD of CP and PP [68.4% (2.7% points above CON) and 95.1% (2.5% points above CON), respectively] whilst PhyG5,000 further increased ATTD PP and also increased ATTD P [54.1% (7.8% points above CON)]; ATTD of Ca tended to be increased in PhyG5,000 vs. CON. Linear dose-response relationships were observed for ATTD of DM, CP, P, Ca, and PP. In addition, fecal excretion of P, and PP linearly reduced and that of Ca and CP tended to linearly reduce with increasing PhyG dose level. No difference was observed for DM intake and milk composition was unaffected except for milk protein which tended to be higher in cows fed PhyG5,000 than CON. In summary, the addition of exogenous phytase at 2,000 FTU/kg or higher to diets of lactating dairy cows improved P, PP, Ca, and CP digestibility and reduced fecal excretion of P, PP, and CP in a dose-dependent manner.

Traditionally, it has been believed that dairy cows are able to fully utilize the phosphorus (P) in feed, including that from plant-derived phytate, because of phytase activity of bacteria in the rumen. However, recent data have shown otherwise. This study investigated the effect of a biosynthetic bacterial 6-phytase supplemented to the diets of midlactating dairy cows on the digestibility and excretion of phosphorus and other key nutrients, over a 19-d experimental period. The experimental diets were commercially relevant in composition and low in phosphorus. At either or both of two tested dose levels (2,000 and 5,000 phytase units (FTU) per kilogram DM in the total diet), the exogenous phytase increased the digestibility and reduced fecal excretion of crude protein (CP), total P, and phytate-P compared with a comparable unsupplemented diet. The increases in CP, PP, and P digestibility were phytase-dose dependent. In addition, at the highest dose level, the phytase tended to increase the protein content of milk. The findings indicate that the use of exogenous phytase can improve P and protein utilization in dairy cows and offers an important approach to optimizing nutrient balance and reducing environmental P and nitrogen (N) pollution from dairy farms.

Ileal and total tract digestibility of energy and nutrients in pig diets supplemented with a novel consensus bacterial 6-phytase variant.

An experiment was conducted to test the hypothesis that increasing levels of a novel phytase increases the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in diets fed to young pigs. A negative control (NC) diet based on corn, soybean meal, and canola meal that contained approximately 0.83% phytate (i.e., 0.23% phytate-bound P) was formulated to be deficient in Ca, P, and standardized ileal digestible amino acids (AA). Five additional diets were formulated by adding 250, 500, 1,000, 2,000, or 4,000 phytase units/kg of the novel phytase to the NC diets. Eighteen ileal-cannulated pigs (17.81 ± 1.71 kg) were allotted to a 6 × 3 incomplete Latin square design with six diets and three 11-day periods. There were three pigs per diet in each period; therefore, there were nine replicate pigs per diet. The initial 5 d of each period was considered an adaptation period to the diet. For each period, fecal samples were collected via anal stimulation on days 6, 7, 8, and 9, whereas ileal digesta were collected on days 10 and 11 using standard procedures. Results indicated that the AID of crude protein, indispensable AA, and dispensable AA was increased (quadratic, P < 0.05) as the concentration of microbial phytase increased in the diets. Dietary inclusion of the novel phytase at 1,000 or 2,000 FTU/kg increased the AID of total AA from 73.7% to 79.8%. Increasing levels of microbial phytase increased (quadratic, P < 0.05) the AID of dry matter and minerals (i.e., Ca, P, K, Mg, Cu) in the diets. Likewise, a linear increase (P < 0.05) in the AID of ash and Na was observed as the inclusion level of phytase increased in the diets. Increasing levels of microbial phytase increased (linear, P < 0.01) the AID of gross energy (GE) and starch in the diets. A quadratic (P < 0.05) increase in the ATTD of ash, Ca, P, K, and Cu in experimental diets was observed as the concentration of microbial phytase increased in the diets. The ATTD of Mg and GE also increased (linear; P < 0.05) as concentration of dietary phytase increased. In conclusion, the novel microbial phytase used in this experiment was effective in increasing the AID of dry matter, GE, starch, minerals, and AA, as well as the ATTD of gross energy and minerals in diets formulated to be deficient in Ca, P, and AA.

The effect of microbial phytase on amino acid (AA) digestibility has been inconsistent, but in many experiments, relatively low levels of phytase were used and it is not known if greater concentrations of phytase are needed to increase AA digestibility. A novel consensus bacterial 6-phytase variant has been recently developed, but it is not known if this phytase results in increased digestibility of AA and other nutrients. Therefore, an experiment was conducted to test the hypothesis that dietary inclusion of increasing levels of the novel phytase (i.e., 0, 250, 500, 1,000, 2,000, and 4,000 phytase units/kg) increases ileal digestibility of AA and total tract digestibility of energy and minerals in diets for growing pigs. In this experiment, it was demonstrated that increasing levels of phytase increased the apparent ileal digestibility of starch, gross energy, minerals, crude protein, and AA, as well as the apparent total tract digestibility of gross energy and minerals. The impact of phytase on AA digestibility is possibly dependent on diet composition, phytate and phytase sources and concentrations, and pig maturity; however, further research is needed to confirm this.

Effects of limestone solubility on the efficacy of a novel consensus bacterial 6-phytase variant to improve mineral digestibility, retention, and bone ash in young broilers fed low-calcium diets containing no added inorganic phosphate.

This study evaluated the effect of limestone solubility on the capacity of a novel consensus bacterial 6-phytase variant (PhyG) to improve phosphorus (P) and calcium (Ca) digestibility, retention, and utilization in low-Ca broiler diets containing no added inorganic phosphate (Pi). Male Ross 308 broilers (n = 1,152) were fed one of 16 experimental diets from 11 to 21 d of age in a randomized complete design (12 birds/cage, 6 cages/treatment). Diets comprised three positive controls (PC3, PC2, and PC1) containing 1.8, 1.2, or 0.6 g/kg MCP-P and 7.7, 7.0, or 6.2 g/kg Ca, respectively, and a negative control (NC) containing no added Pi (4.4 g/kg P; 2.8 g/kg phytate-P) and 5.5 g/kg Ca from either low or high solubility limestone (LSL or HSL, respectively, [with 42% and 97% solubility after 5 min at pH 3.0]), supplemented with 0, 250, 500, 1,000, or 2,000 FTU/kg of PhyG. Fecal samples collected on days 18 to 20 and ileal digesta collected on day 21 were analyzed for titanium dioxide, Ca, P, and phytate (IP6, inositol hexakisphosphate). Tibias (day 21) were analyzed for ash content. Data were analyzed by factorial analysis (2 limestone solubilities × 4 MCP-P levels and 2 limestone solubilities × 5 phytase dose levels) and exponential regression. Increasing dose levels of PhyG resulted in an exponential increase (P < 0.01) in the apparent ileal digestibility (AID) of P, ileal digestible P content of the diet, ileal IP6 content, and IP6 disappearance in birds fed either HSL or LSL diets, but AID Ca and ileal digestible Ca were exponentially increased by the phytase only in HSL diets (P < 0.01). Relative to HSL, the LSL increased AID P, ileal digestible P, and IP6 disappearance (P < 0.05) but reduced AID Ca, ileal digestible Ca, and retainable Ca (P < 0.05), resulting in reduced retainable P and tibia ash. Phytase exponentially increased the apparent total tract digestibility of P, retainable P, and tibia ash in HSL and LSL diets, but at or above 500 FTU/kg values were higher in HSL than LSL (interaction P < 0.05). The findings highlight that phytase dose-response effects on mineral digestibility and utilization are different for high- and low-solubility limestones, and it is therefore recommended to use digestible rather than total Ca content during diet formulation to ensure an optimal balance of Ca and P, especially in low-Ca diets. In diets containing HSL, higher phytase dose levels may be needed to compensate for the low digestible P content of the basal diet.

In broilers, an excess of dietary calcium (Ca) or imbalance with phosphorus (P) can impair mineral digestion and utilization. As a result, diets are being formulated with less Ca, but the quality of the added Ca (that is mainly from limestone) is also important. This study investigated effects of limestone solubility (high [HSL] vs. low [LSL]) on the capacity of a novel consensus bacterial 6-phytase variant, PhyG, to improve P and Ca digestion and utilization in low-Ca diets containing no added inorganic phosphate. Increasing the phytase dose increased ileal P and phytate digestibility and the digestible P content of the diet at 21 d of age regardless of limestone solubility and reduced the negative effects of HSL (relative to LSL). Total tract digestibility of P and Ca, retainable P and Ca, and tibia ash were also increased by phytase, but responses were reduced with LSL relative to HSL. The findings highlight that phytase dose-responses differ in diets containing different limestones and it is therefore recommended to formulate diets based on the content of digestible rather than total Ca to ensure that Ca requirements are met but not exceeded, with optimal phytase efficacy. In diets containing HSL, a higher PhyG dose level is needed to meet the requirement for P.

Microbial phytase reduces basal endogenous loss of calcium in pigs fed diets containing phytate phosphorus at commercial levels.

The objective of this experiment was to test the hypothesis that increasing dietary phytase reduces basal endogenous loss of Ca and increases P balance in pigs. Seventy barrows (initial body weight: 17.66 ± 1.69 kg) were allotted to seven Ca-free diets using a randomized complete block design with two blocks and five pigs per diet in each block. All diets were based on corn, potato protein concentrate, and full-fat rice bran. A positive control (PC) diet was formulated to contain P at the requirement for standardized total tract digestible (STTD) P by 11 to 25 kg pigs. Six negative control (NC) diets were formulated by reducing the provision of digestible P by 0.15% and adding 0, 250, 500, 1,000, 2,000, or 4,000 phytase units/kg diet. Pigs were housed individually in metabolism crates that allowed for total, but separate, collection of urine and feces. Daily feed allowance was 3.0 times the maintenance requirement for metabolizable energy and was divided into two equal meals. Diets were fed for 12 d with the first 5 d considered the adaptation period. Urine collections started on day 6 in the morning and ceased on day 10 in the morning. Fecal markers were also included in the morning meals on day 6 and day 10 and feces were collected according to the marker-to-marker procedure. Results indicated that the apparent total tract digestibility of dry matter was not affected by dietary P or phytase levels. The basal endogenous loss of Ca was not affected by dietary P, but exponentially decreased (P = 0.030) as phytase level increased in the diets. Phosphorus retention (g/d) and standardized total tract digestibility of phosphorus were greater (P < 0.05) in pigs fed the PC diet compared with pigs fed the NC diet with no phytase. The STTD of P exponentially (P < 0.001) increased as phytase level increased in the diets, but because of the lack of Ca, retention of P (% of absorbed) linearly decreased (P = 0.006) as phytase increased. In conclusion, basal endogenous loss of Ca decreased as dietary phytase increased demonstrating that endogenous Ca can be bound to phytate in the intestinal tract of pigs. However, STTD of P increased as phytase level in the diets increased.

Phytate in plant-based ingredients limits the amount of phosphorus available for absorption and can form indigestible complexes with endogenous calcium. However, breakdown of the phytate molecule by phytase increases digestibility of phosphorus and may also reduce endogenous loss of calcium. Therefore, the objective of this experiment was to test the hypothesis that level of phytase influences the utilization of phosphorus and basal endogenous loss of calcium in growing pigs fed calcium-free diets. Results demonstrated that total tract digestibility of phosphorus increased as phytase increased in the diets, but not all of the absorbed P was retained in the pigs due to a lack of calcium. Dietary concentration of phosphorus did not affect basal endogenous loss of calcium, but increasing concentrations of microbial phytase reduced basal endogenous loss of calcium. These results demonstrate that some endogenous calcium is bound to phytate, but including microbial phytase in diets helps release this calcium and therefore reduce endogenous loss of calcium.

Effects of a novel consensus bacterial 6-phytase variant on the apparent ileal digestibility of amino acids, total tract phosphorus retention, and tibia ash in young broilers.

The effect of a novel consensus bacterial 6-phytase variant (PhyG) on apparent ileal digestibility (AID) of amino acids (AA) and phosphorus (P) utilization in young broilers when added to diets with high phytate-P (PP) content without added inorganic phosphate (Pi) and deficient in digestible (dig) AA and metabolizable energy (ME) was investigated. A total of 256 Ross 308 male broilers were assigned to 4 treatments (8 birds/cage, 8 cages/treatment) in a completely randomized design. Treatments comprised a positive control (PC, 2,975 kcal/kg ME, 3.7 g/kg dig P, 2.83 g/kg PP, 8.4 g/kg Ca, 10.6 g/kg dig lysine), a negative control (NC) without added Pi (ME -68 kcal/kg, crude protein -10 g/kg, dig AA -0.1 to -0.4 g/kg, Ca -2.0 g/kg, dig P -2.2 g/kg, Na -0.4 g/kg vs. PC), and NC plus 500 or 1,000 FTU/kg of PhyG. Test diets were corn/soy/rapeseed-meal/rice-bran-based and fed from 5 to 15 d of age. Ileal digesta and tibias were collected on day 15. Excreta was collected during days 12 to 15 to determine P retention. The NC (vs. PC) reduced (P < 0.05) P retention (-10.4% units), tibia ash (-14.3% units), weight gain (-109 g), feed intake (-82 g) and increased FCR (from 1.199 to 1.504), confirming that the NC was extremely deficient in nutrients and energy. Phytase addition to the NC linearly (P < 0.001) improved performance, but did not fully recover it to the level of the PC due to the severe nutrients/energy reduction in NC. Phytase linearly increased P retention (P < 0.001), tibia ash (P < 0.001), AID of dry matter (P < 0.05), nitrogen (P < 0.01), gross energy (P < 0.05), and all 17 individual AA (P < 0.01). At 1,000 FTU/kg, phytase increased (P < 0.05) P retention vs. PC and NC (+14.5 and +24.9% units, respectively) and increased tibia ash vs. NC (+13.8% units), equivalent to PC. The NC decreased AID of Cys, Gly, Thr, and Met vs. PC (P < 0.05). At 1,000 FTU/kg, phytase increased AID of all 17 AA vs. NC (P < 0.01), equivalent to PC. At 1,000 FTU/kg, AID AA responses (above NC) ranged from +4.5% (Met) to +15.0% (Cys), being maximal for essential Thr (+10.4%) and Val (+8.2%) and non-essential Cys (+15.0%) and Gly (+10.4%). The results highlight the efficacy of PhyG at a dose level of 500 to 1,000 FTU/kg in young broilers for improving the ileal digestibility of nitrogen, AA, and energy alongside P retention and tibia ash. The performance data emphasize the need to consider digestible nutrient intake as a response variable in exogenous enzyme studies.

Microbial phytase is widely used in commercial broiler diets to improve digestion of phosphorus (P) and reduce its excretion into the environment. Phytase improves the digestion of phosphorus and other nutrients including amino acids (AA). This study evaluated the effect of a novel consensus bacterial 6-phytase variant (PhyG) added to a nutrient-reduced diet without any added inorganic P on the digestibility of nutrients including P and AA in the ileum of young broilers. Effects on P retention and bone mineralization were also assessed. Compared to an unsupplemented negative control diet, PhyG improved growth performance, P retention, bone mineralization (tibia ash), digestibility of dry matter, nitrogen, gross energy, and all 17 individual AA during 5 to 15 d post-hatch, in a dose-dependent manner (dose range 0 to 1,000 phytase units [FTU] per kilogram of feed). For some AA, the increases in digestibility with PhyG at 1,000 FTU/kg were substantial (cysteine: +15.0%, threonine:+10.4%), and for all AA were equivalent to the responses produced by a nutritionally adequate positive control (unsupplemented) diet. The results demonstrate the efficacy of PhyG to improve AA digestibility alongside growth performance, P retention, and bone mineralization in young broilers.

Influence of a novel consensus bacterial 6-phytase variant on mineral digestibility and bone ash in young growing pigs fed diets with different concentrations of phytate-bound phosphorus.

A 20-d experiment was conducted to test the hypothesis that phytase increases nutrient digestibility, bone ash, and growth performance of pigs fed diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. Within each level of phytate, five diets were formulated to contain 0, 500, 1,000, 2,000, or 4,000 phytase units (FTU)/kg of a novel phytase (PhyG). Three reference diets were formulated by adding a commercial Buttiauxella phytase (PhyB) at 1,000 FTU/kg to diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. A randomized complete block design with 144 individually housed pigs (12.70 ± 4.01 kg), 18 diets, and 8 replicate pigs per diet was used. Pigs were adapted to diets for 15 d followed by 4 d of fecal collection. Femurs were collected on the last day of the experiment. Results indicated that diets containing 0.35% phytate-bound P had reduced (P < 0.01) digestibility of Ca, P, Mg, and K compared with diets containing less phytate-bound P. Due to increased concentration of total P in diets with high phytate, apparent total tract digestible P and bone ash were increased by PhyG to a greater extent in diets with 0.29% or 0.35% phytate-bound P than in diets with 0.23% phytate-bound P (interaction, P < 0.05). At 1,000 FTU/kg, PhyG increased P digestibility and bone P more (P < 0.05) than PhyB. The PhyG increased (P < 0.01) pig growth performance, and pigs fed diets containing 0.35% or 0.29% phytate-bound P performed better (P < 0.01) than pigs fed the 0.23% phytate-bound P diets. In conclusion, the novel phytase (i.e., PhyG) is effective in increasing bone ash, mineral digestibility, and growth performance of pigs regardless of dietary phytate level.

Exogenous enzymes and probiotics alter digestion kinetics, volatile fatty acid content and microbial interactions in the gut of Nile tilapia.

Sustainable aquafeed production requires fishmeal replacement, leading to an increasing use of plant-derived ingredients. As a consequence, higher levels of antinutritional substances, such as non-starch polysaccharides and phytate, are present in aquafeeds, with negative effects on fish performance, nutrient digestibility and overall gut health. To alleviate these negative effects, providing exogenous digestive enzymes and/or probiotics can be an effective solution. In this study, we tested the effect of dietary supplementation of enzymes (phytase and xylanase) and probiotics (three strains of Bacillus amyloliquefaciens) on nutrient digestion kinetics and volatile fatty acid content along the gut, and the distal gut microbiome diversity in Nile tilapia. Chyme volatile fatty content was increased with probiotic supplementation in the proximal gut, while lactate content, measured for the first time in vivo in fish, decreased with enzymes along the gut. Enzyme supplementation enhanced crude protein, Ca and P digestibility in proximal and middle gut. Enzymes and probiotics supplementation enhanced microbial interactions as shown by network analysis, while increased the abundance of lactic acid bacteria and Bacillus species. Such results suggest that supplementation with exogenous enzymes and probiotics increases nutrient availability, while at the same time benefits gut health and contributes to a more stable microbiome environment.

Effect of two phytases at two doses on performance and phytate degradation in broilers during 1-21 days of age.

The effect of two microbial phytases at two dose-levels on performance and apparent ileal digestibility (AID) of nutrients in broilers fed European-type diets was studied. A total of 1,200 d-old Ross 308 male broilers were randomly assigned to 5 treatments with 30 birds/pen and 8 pens/treatment. A nutritionally adequate positive control (PC) diet was tested against 4 experimental diets containing reduced total P, retainable P, Ca and Na as per the recommended nutritional contribution for Buttiauxella phytase (Phy B) at 1,000 FTU/kg (-1.87 g/kg, -1.59 g/kg, -1.99 g/kg and -0.4 g/kg vs. PC, respectively). Experimental diets were supplemented with Phy B at 500 FTU/kg or 1,000 FTU/kg, or Citrobacter phytase (Phy C) at 1,000 FTU/kg or 2,000 FTU/kg. Diets were based on corn, soybean meal, rapeseed meal and sunflower meal and formulated by phase (starter 1-10 d, grower 11-21 d) in crumbled or pelleted form. Overall (d 1-21), at 1,000 FTU/kg, birds fed Phy C exhibited lower BWG (-2.7%), FI (-3.4%) and tibia ash (-2.2%) vs. PC (P < 0.05), and reduced BWG (-3.6%), FI (-3.9%) and tibia ash (-1.8%) vs. Phy B (P < 0.05). Phy B at 1,000 FTU/kg and Phy C at 2,000 FTU/kg maintained performance equivalent to the PC. Digestibility of Ca did not differ among phytase treatments but at 1,000 FTU/kg AID P was greater with Phy B than Phy C (72.3% vs. 62.7%, P < 0.05). Ileal phytate (myo-inositol hexakisphosphate, IP6) digestibility was greatest with Phy B at 1,000 FTU/kg which was higher than Phy C at 1,000 FTU/kg (87.6 vs. 60.6%, P < 0.05). The findings indicate a higher phytate degradation rate of Phy B than Phy C at equivalent dose-level and this is correlated to the performance of the broilers.

Corrigendum to "Functionality of a next generation biosynthetic bacterial 6-phytase in enhancing phosphorus availability to weaned piglets fed a corn-soybean meal-based diet without added inorganic phosphate" [Animal Nutrition 6/1 (2020) 24-30].

[This corrects the article DOI: 10.1016/j.aninu.2019.11.003.].

Safety evaluation of a novel variant of consensus bacterial phytase.

A 90-day subchronic oral toxicity study was conducted to evaluate the safety of a consensus bacterial phytase variant 6-phytase (PhyG) for use as an animal feed additive. This phytase is produced by fermentation with a fungal (Trichoderma reesei) production strain expressing a biosynthetic variant of a consensus bacterial phytase gene assembled via ancestral reconstruction with sequence bias for the phytase from Buttiauxella sp. Rats were administered PhyG daily via oral gavage at dose-levels of 0 (distilled water), 250, 500 or 1000 mg total organic solids (TOS)/kg bodyweight (bw)/day (equivalent to 0, 112,500, 225,000 and 450,000 phytase units (FTU)/kg bw/day, respectively). No test article-related adverse effects were observed. A no-observed-adverse-effect level (NOAEL) for PhyG was established as 1000 mg TOS/kg bw/day, the highest test concentration. Based on this NOAEL and an estimate of broiler consumption determined from the proposed inclusion of the phytase in feed at the maximum recommended level (4000 FTU/kg), a margin of safety value of 1613 was calculated. Results of in vitro genotoxicity testing and in silico protein toxin evaluation further confirmed PhyG to be non-genotoxic and not likely to be a protein toxin upon consumption. These data support the safety of PhyG as an animal feed additive.

Functionality of a next generation biosynthetic bacterial 6-phytase in enhancing phosphorus availability to weaned piglets fed a corn-soybean meal-based diet without added inorganic phosphate.

The utility of a next generation biosynthetic bacterial 6-phytase (PhyG) in restoring bone ash, bone phosphorus (P) content and performance in piglets depleted in P was evaluated. A total of 9 treatments were tested as follows. Treatment 1, a negative control (NC) diet; treatments 2, 3, 4, NC supplemented with 250, 500 or 1,000 FTU/kg of PhyG; treatments 5, 6, NC supplemented with 500 or 1,000 FTU/kg of a commercial Buttiauxella sp phytase (PhyB); treatments 7, 8, 9, NC supplemented with monocalcium phosphate (MCP) to provide 0.7, 1.4 and 1.8 g/kg digestible P, equating to a digestible P content of 1.8, 2.5 and 2.9 g/kg. The latter constituting the positive control (PC) diet with adequate P and calcium (Ca). The NC was formulated without inorganic P (1.1 g digestible P/kg) and reduced in Ca (5.0 g/kg). Additional limestone was added to treatments 7 to 9 to maintain Ca-to-P ratio between 1.2 and 1.3. A total of 162 crossed Pietrain × (Large White × Landrace) 21-d-old piglets (50% males and 50% females) were fed adaptation diets until 42 d old and then assigned to pens with 2 pigs/pen and 9 pens/treatment in a completely randomized block design. Piglets were fed mash diets based on corn and soybean meal ad libitum for 28 d. At the end of the study, one piglet perpen was euthanized and the right feet collected for determination of bone strength, bone ash and mineral content. Compared with the PC, the NC group had reduced average daily gain (ADG) and increased feed conversion ratio (FCR) during all growth phases and overall, and at d 28 (70 d old) NC pigs had bones with reduced ash, Ca and P content (P < 0.05). The PhyG at 250 FTU/kg improved bone ash vs. NC. Increasing PhyG dose linearly or quadratically improved bone ash, ADG and FCR (P < 0.05). At ≥ 500 FTU/kg, both PhyG and PhyB maintained ADG and FCR equivalent to PC. Linear regression analysis was done to compare the measured response parameters to increasing digestible P from MCP. Based on this analysis it was shown that PhyG and PhyB at 1,000 FTU/kg could replace 1.83 and 1.66 g/kg digestible P from MCP in the diet, respectively, on average across metacarpi bone ash, ADG or FCR. These findings suggest that the biosynthetic phytase is highly effective in the tested dietary setting.

The influence of phytase, pre-pellet cracked maize and dietary crude protein level on broiler performance via response surface methodology.

BACKGROUND: The reduction of crude protein levels in diets for broiler chickens may generate economic, environmental and flock welfare and health benefits; however, performance is usually compromised. Whole grain feeding and phytase may improve the utilization of reduced crude protein diets. RESULTS: The effects of pre-pellet cracked maize (0, 15% and 30%) and phytase (0, 750 and 1500 FTU/kg) in iso-energetic maize-soy diets with three levels of crude protein (22%, 19.5% and 17%) were evaluated via a Box-Behnken response surface design. Each of 13 dietary treatments were offered to 6 replicate cages (6 birds/cage) of male Ross 308 broiler chicks from 7 to 28 d post-hatch. Model prediction and response surface plots were generated from experimental data via polynomial regression in R and only significant coefficients were included and discussed in the predicted models. Weight gain, feed intake and FCR were all influenced by pre-pellet cracked maize, phytase and crude protein level, where crude protein level had the greatest influence. Consequently, the reduction from 22% to 17% dietary crude protein in non-supplemented diets reduced weight gain, feed intake, relative gizzard weight, relative gizzard content and relative pancreas weight but improved FCR. However, the inclusion of 30% cracked maize to 17% crude protein diets restored gizzard weight and 1500 FTU phytase inclusion to 17% crude protein diets increased relative gizzard contents and pancreas weights. Cracked maize and phytase inclusion in tandem to 17% crude protein diets increased weight gain, feed intake and FCR; however, this FCR was still more efficient than broilers offered the non-supplemented 22% crude protein diet. Broilers offered the pre-pellet cracked maize and phytase inclusions reduced AME in 22% crude protein diets but improved AME by 2.92 MJ (14.16 versus 11.24 MJ; P < 0.001) in diets containing 17% crude protein. Ileal N digestibility was greater in broilers offered diets with 17% crude protein than those offered the 22% crude protein diet; irrespective of phytase and pre-pellet cracked maize. CONCLUSION: Pre-pellet cracked maize and phytase inclusions will improve the performance of broilers offered reduced crude protein diets.

Increasing the dosing of a Buttiauxella phytase improves phytate degradation, mineral, energy, and amino acid digestibility in weaned pigs fed a complex diet based on wheat, corn, soybean meal, barley, and rapeseed meal1.

This study evaluated the effects of increasing the dose of a 6-phytase from Buttiauxella on phytate degradation, mineral, energy, and AA digestibility in weaned pigs fed complex diets based on wheat, corn, soybean meal, barley, and rapeseed meal. A negative control (NC) diet containing no added inorganic phosphorus (P) and a reduction of 0.1% calcium (Ca) and 36 kcal/kg ME was supplemented with Buttiauxella phytase at 0, 250, 500, 1,000, or 2,000 FTU/kg diet and tested against a nutritionally adequate, positive control (PC) diet. One phytase units (FTU) is the amount of enzyme that liberates 1 micromole of inorganic phosphate per minute from a sodium phytate substrate at pH 5.5 and 37 °C. Barrows (Topigs × Pietrian; initial mean body weight 19.3 kg) were housed individually in metabolic crates and fed the test diets in mash form via 2 equal meals per day for 9 d (fed at 2.5 times the maintenance energy requirement), with 8 replicate pigs per treatment, in 2 experimental runs (total n = 48). After a 3-d adaptation period, urine and feces were collected over 5 d for measurements of apparent total tract digestibility (ATTD) and retention of nutrients. On day 9, pigs were euthanized and ileal digesta collected for measurements of apparent ileal digestibility (AID) of nutrients. Phytase improved (P < 0.05) digestibility of all measured AA except Trp (P < 0.1), and AID P, nitrogen, phytate, ATTD P, Ca versus NC. Increasing phytase dose from 0 (NC) to 2,000 FTU/kg increased AID Lys, Cys, Thr, Val, Ile, Leu, mean AA, P, N, phytate, ATTD P, N, Na, energy, ME, P retention (g/d), and reduced P excretion (g/d) in a linear or exponential manner (P < 0.05). Phytase at 2,000 FTU/kg improved AA digestibility by between +3.1 percentage points (Trp) and +8.8 percentage points (Cys) versus NC (average +6.3 percentage points) (P < 0.05). Phytase inclusion at 2,000 FTU/kg reduced P excretion (g/d) by 57% versus PC (P < 0.05). In conclusion, increasing Buttiauxella phytase in the range of 0 to 2,000 FTU/kg increased phytate degradation, improved AA and P digestibility, and reduced P excretion in weaned pigs fed complex diets.

Productive performance of commercial growing and finishing pigs supplemented with a Buttiauxella phytase as a total replacement of inorganic phosphate.

The objective of this study was to test if a novel phytase from Buttiauxella sp. can replace all added inorganic phosphate in a diet with reduced Ca and metabolizable energy (ME) fed to commercial pigs from 12 kg body weight (BW) until slaughter, whilst maintaining performance and carcass quality parameters. Four dietary treatments were tested in a completely randomized design with 9 replicate pens, each containing 31 mixed sex Newsham Choice pigs. Diets included a positive control (PC) based on corn, soybean meal, wheat middling and bakery meal, meeting all nutrient requirement of pigs; a negative control (NC) excluded inorganic phosphate and with reduced Ca (-0.13%) and ME (-0.15 MJ/kg); and NC supplemented with Buttiauxella phytase at 500 or 1,000 FTU/kg feed. Diets were fed ad libitum in mash form in 5 phases: starter (12 to 25 kg BW), grower 1 (25 to 50 kg BW) and 2 (50 to 75 kg BW), and finisher 1 (75 to 100 kg BW) and 2 (100 kg BW to slaughter). The NC group showed lower (P < 0.05) average daily feed intake (ADFI) and average daily gain (ADG) in starter and grower phases, lower gain to feed ratio (G:F) in starter and grower 1 compared with PC. Pigs receiving the high dose of phytase of 1,000 FTU/kg had improved performance vs. the 500 FTU/kg phytase treatment in starter and grower 1 phase compared with the PC in grower 1 phase. Increasing phytase dose resulted in a linear increase in ADG (12 to 120 kg BW) and G:F (50 to 75 kg BW). A comparison of treatment groups over the full production period from 12 kg BW until slaughter showed that both 500 and 1,000 FTU/kg phytase treatments were able to maintain growth performance and carcass characteristics compared with PC. The application of Buttiauxella phytase could therefore be used as an effective strategy to replace all inorganic phosphate in diets of pigs fed corn, soybean meal, wheat middling and bakery meal based diets from 12 kg BW. An economic analysis showed greater return from both phytase treatments vs. the PC and favored the higher phytase dose at 1,000 FTU/kg vs. the traditional dose of 500 FTU/kg. The latter was mainly related to the improved performance of the higher dose in younger pigs to 75 kg BW.

Phytase in non-ruminant animal nutrition: a critical review on phytase activities in the gastrointestinal tract and influencing factors.

This review focuses on phytase functionality in the digestive tract of farmed non-ruminant animals and the factors influencing in vivo phytase enzyme activity. In pigs, feed phytase is mainly active in the stomach and upper part of the small intestine, and added phytase activity is not recovered in the ileum. In poultry, feed phytase activities are mainly found in the upper part of the digestive tract, including the crop, proventriculus and gizzard. For fish with a stomach, phytase activities are mainly in the stomach. Many factors can influence the efficiency of feed phytase in the gastrointestinal tract, and they can be divided into three main groups: (i) phytase related; (ii) dietary related and (iii) animal related. Phytase-related factors include type of phytase (e.g. 3- or 6-phytase; bacterial or fungal phytase origin), the pH optimum and the resistance of phytase to endogenous protease. Dietary-related factors are mainly associated with dietary phytate content, feed ingredient composition and feed processing, and total P, Ca and Na content. Animal-related factors include species, gender and age of animals. To eliminate the antinutritional effects of phytate (IP6), it needs to be hydrolyzed as quickly as possible by phytase in the upper part of the digestive tract. A phytase that works over a wide range of pH values and is active in the stomach and upper intestine (along with several other characteristics and in addition to being refractory to endogenous enzymes) would be ideal.

A critical review of methodologies used in determination of relative bio-availability ratio of RRR-alpha-tocopheryl acetate and all-rac-alpha-tocopheryl acetate.

Bio-availability of different alpha-tocopherol forms in livestock animals is measured by the increase in plasma or tissue concentrations of alpha-tocopherol after oral administration. It is generally accepted that RRR-alpha-tocopheryl acetate (natural source vitamin E derived from vegetable oil) has a higher bio-availability compared to all-rac-alpha-tocopheryl acetate (synthetic vitamin E, i.e. alpha-tocopherol produced by chemical synthesis). However, different bio-availability ratios have been reported in the literature. The major reason for conflicting results in literature studies was the inability to separate the proportion of alpha-tocopherol originating from test materials, from the proportion of alpha-tocopherol originating from basal dietary ingredients and pre-feeding. This causes significant variability. For bio-availability determination, a baseline or control treatment is essential. The estimation of bio-availability without correction for basal vitamin E status will lead to incorrect interpretation of the results. When using proper methodologies, it is possible to correct for the impact of alpha-tocopherol intake from basal ingredients and alpha-tocopherol originating from pre-feeding, therefore yielding results reflecting the true relative bio-availability of different alpha-tocopherol substances. When reviewing literature data a critical evaluation of the method used in determination of relative bio-availability is recommended.

The impact of low concentrations of aflatoxin, deoxynivalenol or fumonisin in diets on growing pigs and poultry.

In the present review, the quantitative impact of dietary aflatoxin, deoxynivalenol (DON) and fumonisin concentrations on performance of pigs and broilers is evaluated, with special emphasis on low concentrations of these toxins. Also, responses in performance of pigs and broilers to these three toxins are related to their absorption and elimination kinetics. By applying simple linear regression, information from many literature sources is integrated and condensed into, for example, estimates of depression in rates of weight gain, relative to non-contaminated diets, with increasing toxin concentrations. It was estimated that with each mg/kg increase of aflatoxin in the diet, the growth rate would be depressed by 16 % for pigs and 5 % for broilers. For DON, with each mg/kg increase in the diet, the growth depression was estimated at about 8 % for pigs, while broilers showed no response to DON concentrations below 16 mg/kg. Fumonisin showed the lowest impact on growth performance; with each mg/kg increase, the depression in growth rate was estimated at 0.4 and 0.0 % for pigs and broilers, respectively. Dietary concentrations that cause a 5 % reduction in growth rate were estimated at 0.3 and 1.0 mg/kg for aflatoxin for pigs and broilers, respectively; 1.8 and 0.6 mg/kg for pure and naturally contaminated DON for pigs, respectively; 21 and 251 mg/kg for fumonisin for pigs and broilers, respectively.