Evaluation of the effects of pharmacological zinc oxide and phosphorus source on weaned piglet growth performance,plasma minerals and mineral digestibility

Crossbred pigs (n=720; average age=28±3 days and weight=9.5±0.3 kg) were used in a 20-day trial in order to determine the influence of phosphorus (P) source and various doses of pharmacological zinc (Zn) on growth performance, plasma minerals and mineral digestibility. Pigs (five intact males and five females per pen) were randomly allotted to treatments in a 3×3 factorial arrangement with three sources of dietary P (4.5 g/kg digestible P, 4.5 g/kg digestible P plus 2500 phytase units (FTU)/kg, or 5.5 g/kg digestible P) and three dietary levels of supplemental Zn (0, 1750 or 3500 mg/kg) from ZnO (82% Zn) with eight pens per treatment. Diets were formulated to exceed all nutrient requirements, including calcium (Ca), P and Zn from day 0 to 20. Zn supplementation increased (quadratic P<0.05) average daily feed intake. There was a significant Zn level×P source interaction on average daily gain and feed conversion ratio (FCR). Pigs fed 4.5 g/kg digestible P without or with 2500 FTU/kg phytase gained more per day (quadratic P<0.05) and had better FCR (quadratic P<0.05) when they were fed 1750 mg/kg supplemental Zn. However, pigs fed 5.5 g/kg digestible P gained more per day (linear P<0.05) and were more efficient (linear P<0.05) when they were fed 3500 mg/kg supplemental Zn. Plasma Zn and Zn digestibility increased (linear P<0.05) as pharmacological Zn supplementation increased from 0 to 3500 mg/kg, irrespective of P source. However, Ca, P, sodium (Na), potassium (K) and copper (Cu) digestibility were reduced (P<0.05) as pharmacological Zn supplementation increased, and this was mitigated or exacerbated by the supplementation of 5.5 g/kg digestible P or phytase. In conclusion, increasing the dietary inclusion of pharmacological Zn may impact growth performance in young pigs through the interaction with minerals such as Ca, P, Na and K. Pharmacological Zn may reduce Na or K digestibility and indirectly reduce water secretion into the lumen, resulting in an increase in faecal dry matter as pharmacological Zn supplementation in the diet increased.     

Dietary effects on inositol phosphate breakdown in the crop of broilers

The effect of diets differing in enzyme supplements, mineral phosphorus (P) and microwave wheat treatment on phytate hydrolysis and lower inositol phosphate isomers (InsPs) appearance in broiler crops was studied. The broilers (16- and 15-day-old) were assigned to 48 pens of 15 or 20 birds each (n = 8 pens per treatment) in Experiments 1 and 2, respectively. In Experiment 1, birds received a low-P wheat-soybean meal diet where the wheat was either microwave treated or not. These diets were offered without further supplementation or with added phytase (500 FTU/kg diet), alone or in combination with a xylanase (16,000 BXU/kg diet). In Experiment 2, two maize-soybean meal-based diets were fed, without or with monocalcium phosphate supplementation. Furthermore, these diets were offered without further supplementation or with phytase at 500 or 12,500 FTU/kg diet. On day 23 or 24 (Experiments 1 and 2, respectively), crop digesta were pooled per pen, freeze-dried and analysed for InsPs and the marker TiO₂. Microwaving reduced the intrinsic phytase activity and InsP₆ hydrolysis, but increased the concentration of Ins(1,2,3,4,5)P₅ and Ins(1,2,4,5,6)P₅ in the digesta of crop (Experiment 1). Microwave treatment significantly interacted with enzyme supplementation for Ins(1,2,5,6)P₄ concentration, indicating a synergistic effect of intrinsic and supplied phytase in the crop. Xylanase tended to support phytase hydrolysis in diets with microwave-treated wheat. Phytase addition increased InsP₆ hydrolysis up to 79% (Experiment 2). Thus, wheat phytase activity can cause high InsP₆ hydrolysis in the crop. Treatment differences in lower InsPs indicated that hydrolysis of the first InsP₆ phosphate group is not the only step in the degradation cascade in the crop of broilers that is influenced by dietary factors.     

Meta-analysis of phosphorus utilization by growing pigs: effect of dietary phosphorus,calcium and exogenous phytase

Optimizing phosphorus (P) utilization in pigs requires improving our capacity to predict the amount of P absorbed and retained, with the main modulating factors taken into account, as well as precisely determining the P requirements of the animals. Given the large amount of published data on P utilization in pigs, a meta-analysis was performed to quantify the impact of the different dietary P forms, calcium (Ca) and exogenous phytases on the digestive and metabolic utilization criteria for dietary P in growing pigs. Accordingly, the amount of phytate P (PP) leading to digestible P (g/kg) was estimated to be 21%, compared with 73% for non-phytate P (NPP) from plant ingredients and 80% for NPP from mineral and animal ingredients (P < 0.001). The increase in total digestible dietary P following the addition of microbial phytase (PhytM) from Aspergillus niger (P < 0.001) was curvilinear and about two times higher than the increase following the addition of plant phytase, which leads to a linear response (P < 0.001). The response of digestible P to PhytM also depends on the amount of substrate, PP (PhytM² × PP, P < 0.001). The digestibility of dietary P decreased with dietary Ca concentration (P < 0.01) independently of phytase but increased with body weight (BW, P < 0.05). Although total digestible dietary P increased linearly with total NPP concentration (P < 0.001), retained P (g/kg), average daily gain (ADG, g/day) and average daily feed intake (ADFI, g/day) increased curvilinearly (P < 0.001). Interestingly, whereas dietary Ca negatively affected P digestibility, the effect of dietary Ca on retained P, ADG and ADFI depended on total dietary NPP (NPP × Ca, P < 0.01, P < 0.05 and P < 0.01, respectively). Increasing dietary Ca reduced retained P, ADG and ADFI at low NPP levels, but at higher NPP concentrations it had no effect on ADG and ADFI despite a positive effect on retained P. Although the curvilinear effect of PhytM on digestible P increased with PP (P < 0.001), this effect was lessened by total NPP for ADG and ADFI (PhytM × NPP and PhytM² × NPP, P < 0.05) and depended on both total NPP and Ca for retained P (PhytM² × NPP × Ca, P < 0.01). This meta-analysis improves our understanding of P utilization, with major modulating factors taken into account. The information generated will be useful for the development of robust models to formulate environmentally friendly diets for growing pigs.     

Effects of mineral and rapeseed phosphorus supplementation on phytate degradation in dairy cows

The objective of this study was to evaluate the effects of diet composition on phytate (InsP₆) degradation in dairy cows. In Experiment 1, four diets that differed in the amount and source of phosphorus (P) were fed to 24 lactating cows in a 4 × 4 Latin Square design. The control diet (Diet C) contained 4.18 g P/kg dry matter (DM). Diet MP contained additional mineral P (5.11 g P/kg DM), Diet RS contained rapeseed and rapeseed meal as organic P sources (5.26 g P/kg DM) and Diet RSM contained rapeseed meal and rapeseed oil (5.04 g P/kg DM). Total P (tP) and InsP₆ excretion in faeces were measured. In Experiment 2, we used a rumen simulation technique (Rusitec) to estimate ruminal disappearance of tP and InsP₆ from Diets C, MP and RSM. In Experiment 1, tP concentration in faeces increased with tP intake and was highest for Diets RS and RSM. The source of supplemented P had no influence on tP digestibility, but tP digestibility was reduced for Diets MP, RS and RSM in comparison to that for Diet C. InsP₆ disappearance decreased in Diet MP (85.0%) and increased in Diets RS (92.7%) and RSM (94.0%) compared to that in Diet C (90.0%). In Experiment 2, P source influenced ruminal tP disappearance (Diet MP, 78.6%; Diet RSM, 75.3%). InsP₆ disappearance for Diet C (98.1%) was higher than that for Diets MP (95.6%) and RSM (94.9%). The results confirmed the high potential of ruminants to degrade InsP₆, but differences in diet composition influenced InsP₆ disappearance. Further studies of the site of InsP₆ degradation are required to understand the relevance of InsP₆ degradation for the absorption of P.     

Effect of microbial phytase on apparent ileal amino acid digestibility of phosphorus-adequate diets in growing pigs

Six ileally cannulated pigs (mean initial body weight 34.8 kg) were used to study the effect of microbial phytase on apparent ileal digestibility of P, total N and amino acids. Three P-adequate diets (digestible P concentration 2.3 g kg(- )l) containing barley (B), soyabean meal (S) or a mixture of the two (BS) with or without phytase supplement (1000 FTU x kg(-1)) were fed to pigs using a 6 x 6 Latin square design. The addition of phytase increased (p < 0.05) apparent ileal P digestibility of diets B, S and BS by 16.5, 19.2 and 19.2%, respectively. There was no effect of phytase on the ileal digestibility of total N. Apparent ileal digestibility of amino acids tended to increase in the BS diet supplemented with phytase (mean improvement of 2.2%); but no significant difference was found for any amino acid as compared with the unsupplemented diet. To asses the additivity of apparent amino acid digestibility, the determined values for the BS diet were compared to those calculated from digestibilities found in diets B and S. There were no significant differences between the determined and calculated values. It is concluded that the addition of microbial phytase to P-adequate diets does not affect ileal amino acid digestibility in growing pigs and that the apparent amino acid digestibility values determined in single ingredients may be additive when included into a complex diet.     

Effects of rapeseed meal fiber content on phosphorus and calcium digestibility in growing pigs fed diets without or with microbial phytase

The optimization of dietary phosphorus (P) and calcium (Ca) supply requires a better understanding of the effect of dietary fiber content of co-products on the digestive utilization of minerals. This study was designed to evaluate the effects of dietary fiber content from 00-rapeseed meal (RSM) on P and Ca digestibility throughout the gastrointestinal tract in growing pigs fed diets without or with microbial phytase. In total, 48 castrated male pigs (initial BW=36.1±0.4 kg) were housed in metabolic crates for 29 days. After an 8-day adaptation period, pigs were allocated to one of the eight treatments. The impact of dietary fiber was modulated by adding whole RSM (wRSM), dehulled RSM (dRSM) or dRSM supplemented with 4.5% or 9.0% rapeseed hulls (dRSMh1 and dRSMh2). Diets contained 0 or 500 phytase unit of microbial phytase per kg. From day 14 to day 23, feces and urine were collected separately to determine apparent total tract digestibility (ATTD) and apparent retention (AR) of P and Ca. At the end of the experiment, femurs and digestive contents were sampled. No effect of variables of interest was observed on growth performance. Microbial phytase increased ATTD and AR of P (P<0.001) but the P equivalency with the wRSM diet was lower than expected. Moreover, stomach inorganic P (iP) solubility was improved by microbial phytase (P<0.001). The ATTD of Ca was not affected by microbial phytase which increased AR of Ca and femur characteristics (P<0.05). Ileal recovery of P was not affected by microbial phytase but cecal recovery was considerably reduced by microbial phytase (P<0.001). The decrease in digesta pH between the distal ileum and cecum (7.6 v. 5.9) enhanced the solubility of iP and may have improved its absorption, as supported by the negative relationship between soluble iP and pH (R²=0.40, P<0.001 without microbial phytase and R²=0.24, P=0.026 with microbial phytase). The inclusion of hulls improved the solubility of iP (P<0.05). In conclusion, dehulling does not largely increase nutrient digestibility although dRSM seems to improve the efficacy of microbial phytase in releasing phosphate in the stomach. Moreover, dietary fiber may affect solubilization process in the cecum which potentiates the effect of microbial phytase on P digestibility.     

Muscarinic Receptor-Mediated Inositol 1,4,5-Trisphosphate Formation in SH-SY5Y Neuroblastoma Cells Is Regulated Acutely by Cytosolic Ca2+ and by Rapid Desensitization

Abstract: Stimulation of muscarinic receptors expressed in SH-SY5Y human neuroblastoma cells resulted in a complex profile of inositol 1,4,5-trisphosphate (InsP₃) accumulation, with a dramatic increase (six- to eightfold) over the first 10 s (the “peak” phase) and subsequently, from ～60 s onward, maintained at a lower but sustained level (the “plateau” phase). Chelation of extracellular Ca²⁺ with EGTA or inhibition of Ca²⁺ channels with Ni²⁺ showed that the plateau phase was dependent upon Ca²⁺ entry. Furthermore, use of thapsigargin and EGTA to discharge and sequester Ca²⁺ from intracellular stores revealed that Ca²⁺ from this source was capable of supporting the peak phase of the InsP₃ response. Carbachol-stimulated phosphoinositidase C activity in permeabilized SH-SY5Y cells was also shown to be highly dependent on free Ca²⁺ concentration (20–100 nM) and suggests that under normal conditions, InsP₃ formation is enhanced by increases in cytosolic free Ca²⁺ concentration that accompany muscarinic receptor activation. Measurement of carbachol-stimulated total inositol phosphate accumulation in the presence of Li⁺ indicated that the initial rate of phosphoinositide hydrolysis (from 0 to 30 s) was about fivefold greater than that from 30 to 300 s. This rapid but partial desensitization of receptor-mediated phosphoinositide hydrolysis provides strong evidence for the mechanism underlying the changes in InsP₃ accumulation over this time. Because very similar data were obtained in Chinese hamster ovary cells transfected with human m3 receptor cDNA, we suggest that although increases in cytosolic free Ca²⁺ concentration amplify InsP₃ formation during stimulation of m3 muscarinic receptors, the primary factor that governs the profile of InsP₃ accumulation is rapid, but partial, desensitization. Such desensitization does not appear to be mediated by changes in cytosolic Ca²⁺ or protein kinase C activity.     

Effects of β‐xylanase and 6‐phytase on digestibility,trace mineral utilisation and growth in juvenile red tilapia,Oreochromis niloticus (Linnaeus, 1758) x O. mossambicus (Peters, 1852), fed declining fishmeal diets

In response to the global sustainability drive to lower fishmeal (FM) inclusion in aquatic feeds, exogenous enzymes can improve nutrient digestibility in monogastric plant‐based diets. A 80‐day experiment was conducted to evaluate the combined effects of xylanase and phytase on digestibility, trace mineral utilisation and growth in juvenile red tilapia, Oreochromis niloticus x O. mossambicus, (48.8 g ± 13.9; μ ± STD) fed declining FM diets. Basal diets were formulated to contain 0, 3 and 5% FM with and without xylanase (0.385 g kg^?1) and phytase (0.075 g kg^?1), forming six treatments. Each treatment was randomly assigned to four replicates, 20 fish tank^?1; mean water temperature 28.98 ± 0.73°C. Although the size of the effects was modest, growth performances (feed intake, FCR, growth rate) decreased with lower FM levels (P < 0.05) but improved with enzyme supplementation. Enzyme supplementation increased P digestibility and trace mineral uptake (P < 0.05), but no effects were seen on protein digestibility and N retention. Nevertheless, tilapia fed the enzyme‐supplemented 3% FM and control 5% FM diets performed comparably (P < 0.05). This potentially justifies a 2% FM reduction for tilapia diets using exogenous xylanase and phytase without significant effects on digestibility, trace mineral utilisation and growth.     

Effects of phytase and 25-hydroxyvitamin D3 inclusions on the performance, mineral balance and bone parameters of grower-finisher pigs fed low-phosphorus diets

Two experiments, a performance experiment and a mineral balance study, were conducted on grower-finisher pigs (42 to 101 kg live weight) to investigate the effects of Peniophora lycii phytase enzyme and 25-hydroxyvitamin D3 (25-OHD3) on growth performance, carcass characteristics, nutrient retention and excretion, and bone and blood parameters. The two experiments were designed as a 2 × 2 factorial (two levels of phytase and two levels of 25-OHD3). The four diets were T1, low-phosphorous diet; T2, T1 + phytase; T3, T1 + 25-OHD3 and T4, T1 + phytase + 25-OHD3 diet. In all, 25 μg of 25-OHD3 was used to replace 1000 IU of vitamin D3 in diets T3 and T4. Diets were pelleted (70°C) and formulated to contain similar concentrations of energy (13.8 MJ DE/kg), lysine (9.5 g/kg) and digestible phosphorus (P; 1.8 g/kg). Neither the inclusion of phytase nor 25-OHD3 in the diet had any effect on pig performance. There was an interaction between phytase and 25-OHD3 on calcium (Ca) and P retention (P < 0.01) and on the apparent digestibility of ash (P < 0.01), P (P < 0.001) and Ca (P < 0.001). Pigs offered phytase diets only, had a higher retention of Ca and P and digestibility of ash (P < 0.01), P (P < 0.001) and Ca (P < 0.01) compared with pigs offered unsupplemented diets. However, when the combination of phytase and 25-OHD3 were offered, no effects were detected compared with 25-OHD3 diets only. Pigs fed phytase diets had higher bone ash (P < 0.01), bone P (P < 0.01) and bone Ca (P < 0.05) concentrations compared with pigs offered non-phytase diets. In conclusion, pigs offered phytase diets had a significantly increased bone ash, Ca and P than pigs offered unsupplemented phytase diets. However, there was no advantage to offering a combination of phytase and 25-OHD3 on either bone strength or mineral status compared to offering these feed additives separately.     

Improvement in overall performance of Catla catla fingerlings fed phytase included low cost plant by products-based diet

Phytic acid’s presence in low-cost Moringa by-products effect the availability of important nutrients, diminishing the fish quality and blood composition in fish. Phytate having chelating effects with nutrients and minerals, can be reduced by the supplementation of phytase enzyme. Without the use of enzyme, plant meal may cause water pollution and decrease the fish health that results in higher culture cost. Therefore, current study was designed to check improvement in overall performance of Catla catla fingerlings fed Moringa by product-based diets supplemented with phytase (0, 300, 600, 900, 1200 and 1500, FTU/kg). All diets were integrated with non-digestible marker (Cr₂O₃) at the rate of 1%. The fingerlings were fed couple of times a day (4% of live wet weight). Results showed significant (p < 0.05) improvement in nutrient digestibility (i.e. EE, CP and GE), carcass composition and hematological parameters (i.e. RBCs, PLT and Hb) at 900, FTU/kg of phytase in contrast with other treatments. Moreover, phytase addition improves the water quality by reducing the nutrients leaching through feces at low cost. Current results indicated that, using mixture of Moringa seed meal and Moringa leaf meal based diet supplemented with phytase at 900, FTU/kg concentration is the most optimum level to develop a cost-effective as well as eco-friendly fish feed with maximum absorption of important nutrients and minerals in fish body resultantly high higher fish performance.     

the effect of the combination of microbial phytase and amino acid supplementation of diets for finishing pigs on p and n excretion and carcass quality

The objective of this study was to evaluate the effect of a combined low-protein, low-phosphorus diet supplemented with limiting amino acids and microbial phytase on performance, nutrient utilization and carcass characteristics of late-finishing barrows. 4?×?8 crossbreed barrows were continuously housed in metabolism cages from 70?–?110?kg BW and were fed diets, either conventional (A) or protein reduced (B) or protein and phosphorus reduced diets (C) based on barley, maize and soybean meal. Diet A (positive control) contained in air dry matter 13% and 10% CP as well as 0.49% and 0.42% P at growth phases I (70?–?100?kg BW) or II (100?–?110?kg BW), respectively. Diet B was low in CP (11.3%, 8.4%), diet C low in CP and low in P (CP: as B, P: 0.36%, 0.30%). To diet B the limiting amino acids lysine, methionine, threonine and trypthophan were added to meet the levels in diet A. To diet C the limiting amino acids and 800 FTU/kg Aspergillus-phytase were supplemented. At the end of the balance periods the barrows were slaughtered, the carcasses scored and loin chops, ham and Phalanx prima IV were analysed for nutrients and minerals. The CP or P reduction in diets B and C did not generally negatively affect growth, feed efficiency, absolute nitrogen retention or overall carcass performances of the pigs. With the low CP diets B and C, N excretion per unit BWG was decreased by about 23%. The addition of microbial phytase (diet C) increased apparent total tract digestibility of P by about 20%. In spite of 30% reduction of P intake (diet C), the absolute P retention related to 1?kg BW did not differ between treatments. Thus, phytase supplementation in diet C reduced P excretion per unit BWG by about 33%. Phytase raised apparent digestibility of Zn by about 20% but not Ca digestibility. Generally the carcass traits and meat characteristics were not affected by any of the diet strategies. Mineralization of the Phalanx prima IV was also similar in all treatment groups. However, phytase supplementation led to significantly increased zinc concentration in bones (25%). In contrast, Fe incorporation into the Phalanx prima IV was not affected. In general, the feeding regimen introduced in this experiment offers substantial benefits in maintaining a sustainable environmental-friendly pork production even at the stage of late-finishing barrows.     

The effect of high and low dietary crude protein and inulin supplementation on nutrient digestibility, nitrogen excretion, intestinal microflora and manure ammonia emissions from finisher pigs

A 2 × 2 factorial experiment was performed to investigate the interaction between a high- and low-crude-protein (CP) diet (200 v. 140 g/kg) and inulin supplementation (0 v. 12.5 g/kg) on nutrient digestibility, nitrogen (N) excretion, intestinal microflora, volatile fatty acid (VFA) concentration and manure ammonia emissions from 24 boars (n = 6, 74.0 kg live weight). The diets were formulated to contain similar concentrations of digestible energy and lysine. Pigs offered the high-CP diets had a higher excretion of urinary N (P < 0.001), faecal N (P < 0.01) and total N (P < 0.001) than the pigs offered the low-CP diets. Inulin supplementation increased faecal N excretion (P < 0.05) and decreased the urine N : faeces N ratio (P < 0.05) compared with the inulin-free diets. There was no effect (P > 0.05) of dietary treatment on N retention. There was an interaction (P < 0.05) between dietary CP concentration and inulin supplementation on caecal Enterobacteria spp. Pigs offered the diet containing 200 g/kg of CP plus inulin decreased the population of Enterobacteria spp. compared to those with the inulin-supplemented 140 g/kg CP diet. However, CP level had no significant effect on the population of Enterobacteria spp. in the unsupplemented diets. Inulin supplementation increased caecal Bifidobacteria (P < 0.01) compared with the inulin-free diets. There was no effect of inulin supplementation on VFA concentration or intestinal pH (P > 0.05). Pigs offered the 200 g/kg CP diets had higher (P < 0.05) manure ammonia emissions from 0 to 240 h of storage than pigs offered the 140 g/kg CP. In conclusion, inulin supplementation resulted in an increase in Bifidobacteria concentration and a reduction in Enterobacteria spp. at the high CP level indicating that inulin has the ability to beneficially manipulate gut microflora in a proteolytic environment.     

Sparing effect of microbial phytase on zinc supplementation in maize-soya-bean meal diets for chickens

The experiment was conducted to evaluate the sparing effect of microbial phytase on the need for dietary zinc supplementation in chicks. A maize–soya-bean meal basal diet, containing 33 mg of zinc and 16 mg of copper per kg, supplemented with 0, 6, 12, 18, 24, 30 or 60 mg of zinc as sulphate per kg or with 250, 500, 750 or 1000 units (FTU) of microbial phytase (3-phytase from Aspergillus niger, Natuphos®) per kg was given to 1-day-old chicks for 20 days. Sixteen chicks placed in individual cages were assigned to each diet except the unsupplemented basal diet which was assigned to 32 cages. Actual range of phytase supplementation was 280 to 850 FTU per kg diet. Growth performance was not affected by microbial phytase. Chicks given the unsupplemented basal diet and the basal diet supplemented with 60 mg of zinc per kg displayed similar performance. Bone weight, bone ash, liver weight and liver dry matter were independent (P > 0.1) of zinc and phytase supplementations. Plasma, bone and liver zinc concentrations increased linearly (P < 0.001) and quadratically (P < 0.001; P < 0.001 and P < 0.05, respectively) with zinc added. Plasma zinc tended to increase linearly (P = 0.07) and bone zinc increased linearly (P < 0.01) with phytase added but no quadratic response was detected (P > 0.1). Liver zinc was unresponsive to phytase added (P > 0.1). Liver copper decreased linearly (P < 0.001) and quadratically (P < 0.01) with zinc supplementation. Mathematical functions were fitted to the responses of plasma and bone zinc to zinc and phytase added and used to calculate zinc equivalency values of phytase. The models included a linear plateau response to zinc added and a linear response to phytase added. In diets without phytase, plasma and bone zinc concentrations were maximised for a dietary zinc concentration of 55 and 51 mg/kg, respectively. Over the range of 280 to 850 FTU, 100 FTU was equivalent to 1 mg of zinc as sulphate. Consequently, in a maize–soya-bean meal chicken diet formulated to contain 60 mg zinc per kg, zinc ingested, and in turn, zinc excreted may be reduced by around 10% if the diet contains 500 FTU as Natuphos® per kg.     

Jodmangel bei Wiederkäuern

Effects of a supplemental Aspergillus niger‐phytase on digestibility and utilization of dietary phosphorus (P) were studied in three experiments with rainbow trout. P concentration in the diets was 4.8 and 5.8 g/kg DM, respectively. The P contained in the diet originated solely from plants, mainly soy‐products. Digestibility of P was studied using the stripping method and hydrochloride insoluble ash as marker. Utilization was studied in growth trials by use of the comparative body analysis.

At a water temperature of 15°C, both digestibility and utilization of P were increased from 25 to 57% and from 17 to 49%, respectively when 1000 U/kg phytase were supplemented. Feed consumption and gain of trout were significantly increased. At a water temperature of 10°C, utilization of P was also increased from 6 to 25%. However, feed consumption and gain of trout were very low at this water temperature and not influenced by the supplemental phytase.     

Effect of phytase supplementation on phosphorus digestibility in low-phytate barley fed to finishing pigs

Forty crossbred barrows (Camborough 15 Line female×Canabred sire) weighing an average of 79.6±8.0?kg were used in a factorial design experiment (5 barleys×2 enzyme levels) conducted to determine the effects of phytase supplementation on nutrient digestibility in low-phytate barleys fed to finishing pigs. The pigs were assigned to one of 10 dietary treatments comprised of a normal 2-rowed, hulled variety of barley (CDC Fleet, 0.26% phytate) or 2 low-phytate hulled genotypes designated as LP422 (0.14% phytate) and LP635 (0.09% phytate). A normal, hulless barley (CDC Dawn, 0.26% phytate) and a hulless genotype designated as LP422H (0.14% phytate) were also included. All barleys were fed with and without phytase (Natuphos 5000 FTU/kg). The diets fed contained 98% barley, 0.5% vitamin premix, 0.5% trace mineral premix, 0.5% NaCl and 0.5% chromic oxide but no supplemental phosphorus. The marked feed was provided for a 7-day acclimatization period, followed by a 3-day faecal collection. In the absence of phytase, phosphorus digestibility increased substantially (P<0.05) as the level of phytate in the barley declined. For the hulled varieties, phosphorus digestibility increased from 12.9% for the normal barley (0.26% phytate) to 35.3 and 39.8% for the two low-phytate genotypes (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 9.2% for the normal barley (0.26% phytate) to 34.7% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In contrast, when phytase was added to the diet, there was little difference in phosphorus digestibility between pigs fed normal barley and those fed the low-phytate genotypes (significant barley×enzyme interaction, P=0.01). For the hulled varieties, phosphorus digestibility was 50.1% for the barley with the normal level of phytate (0.26% phytate) compared with 51.1 and 52.4% for the varieties with 54 and 35% of the normal level of phytate (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 47.1% for the normal barley (0.26% phytate) to 54.4% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In conclusion, both supplementation with phytase and selection for low-phytate genotypes of barley were successful in increasing the digestibility of phosphorus for pigs. Unfortunately, the effects did not appear to be additive. Whether or not swine producers will choose low-phytate barley or supplementation with phytase as a means to improve phosphorus utilization, will likely depend on the yield potential of low-phytate barley and the additional costs associated with supplementation with phytase.     

Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells

The inositol (1,4,5)-trisphosphate receptor (InsP₃R) mediates Ca²⁺ release from intracellular stores in response to generation of second messenger InsP₃. InsP₃R was biochemically purified and cloned, and functional properties of native InsP₃-gated Ca²⁺ channels were extensively studied. However, further studies of InsP₃R are obstructed by the lack of a convenient functional assay of expressed InsP₃R activity. To establish a functional assay of recombinant InsP₃R activity, transient heterologous expression of neuronal rat InsP₃R cDNA (InsP₃R-I, SI− SII+ splice variant) in HEK-293 cells was combined with the planar lipid bilayer reconstitution experiments. Recombinant InsP₃R retained specific InsP₃ binding properties (K _d = 60 nM InsP₃) and were specifically recognized by anti–InsP₃R-I rabbit polyclonal antibody. Density of expressed InsP₃R-I was at least 20-fold above endogenous InsP₃R background and only 2–3-fold lower than InsP₃R density in rat cerebellar microsomes. When incorporated into planar lipid bilayers, the recombinant InsP₃R formed a functional InsP₃-gated Ca²⁺ channel with 80 pS conductance using 50 mM Ba²⁺ as a current carrier. Mean open time of recombinant InsP₃-gated channels was 3.0 ms; closed dwell time distribution was double exponential and characterized by short (18 ms) and long (130 ms) time constants. Overall, gating and conductance properties of recombinant neuronal rat InsP₃R-I were very similar to properties of native rat cerebellar InsP₃R recorded in identical experimental conditions. Recombinant InsP₃R also retained bell-shaped dependence on cytosolic Ca²⁺ concentration and allosteric modulation by ATP, similar to native cerebellar InsP₃R. The following conclusions are drawn from these results. (a) Rat neuronal InsP₃R-I cDNA encodes a protein that is either sufficient to produce InsP₃-gated channel with functional properties identical to the properties of native rat cerebellar InsP₃R, or it is able to form a functional InsP₃-gated channel by forming a complex with proteins endogenously expressed in HEK-293 cells. (b) Successful functional expression of InsP₃R in a heterologous expression system provides an opportunity for future detailed structure–function characterization of this vital protein.     

Effect of phytase supplementation on phosphorus digestibility in low-phytate barley fed to finishing pigs

Forty crossbred barrows (Camborough 15 Line female x Canabred sire) weighing an average of 79.6 +/- 8.0 kg were used in a factorial design experiment (5 barleys x 2 enzyme levels) conducted to determine the effects of phytase supplementation on nutrient digestibility in low-phytate barleys fed to finishing pigs. The pigs were assigned to one of 10 dietary treatments comprised of a normal 2-rowed, hulled variety of barley (CDC Fleet, 0.26% phytate) or 2 low-phytate hulled genotypes designated as LP422 (0.14% phytate) and LP635 (0.09% phytate). A normal, hulless barley (CDC Dawn, 0.26% phytate) and a hulless genotype designated as LP422H (0.14% phytate) were also included. All barleys were fed with and without phytase (Natuphos 5000 FTU/kg). The diets fed contained 98% barley, 0.5% vitamin premix, 0.5% trace mineral premix, 0.5% NaCl and 0.5% chromic oxide but no supplemental phosphorus. The marked feed was provided for a 7-day acclimatization period, followed by a 3-day faecal collection. In the absence of phytase, phosphorus digestibility increased substantially (P < 0.05) as the level of phytate in the barley declined. For the hulled varieties, phosphorus digestibility increased from 12.9% for the normal barley (0.26% phytate) to 35.3 and 39.8% for the two low-phytate genotypes (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 9.2% for the normal barley (0.26% phytate) to 34.7% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In contrast, when phytase was added to the diet, there was little difference in phosphorus digestibility between pigs fed normal barley and those fed the low-phytate genotypes (significant barley x enzyme interaction, P = 0.01). For the hulled varieties, phosphorus digestibility was 50.1% for the barley with the normal level of phytate (0.26% phytate) compared with 51.1 and 52.4% for the varieties with 54 and 35% of the normal level of phytate (0.14 and 0.09% phytate respectively). For the hulless varieties, phosphorus digestibility increased from 47.1% for the normal barley (0.26% phytate) to 54.4% for the hulless variety with 54% of the normal level of phytate (0.14% phytate). In conclusion, both supplementation with phytase and selection for low-phytate genotypes of barley were successful in increasing the digestibility of phosphorus for pigs. Unfortunately, the effects did not appear to be additive. Whether or not swine producers will choose low-phytate barley or supplementation with phytase as a means to improve phosphorus utilization, will likely depend on the yield potential of low-phytate barley and the additional costs associated with supplementation with phytase.     

Bone mineralisation of weaned piglets fed a diet free of inorganic phosphorus and supplemented with phytase,as assessed by dual-energy X-ray absorptiometry

Sixteen female piglets (58 d of age, 16.8 ± 0.8 kg body weight [BW]) were assigned to two groups (n = 8) and received until day 100 of age (50.3 ± 1.2 kg BW) ad libitum either a diet with a standard (diet C) or low (diet L) total phosphorus (P) content (5.38 and 4.23 g/kg, respectively). Diet C was supplemented with mineral P (1.15 g/kg) and did not contain microbial phytase. Diet L did not contain any inorganic P but 750 FTU/kg of microbial phytase. Despite these treatments, both diets were composed with the same ingredients. Body mineralisation of each gilt was assessed by determining the bone mineral content (BMC), area bone mineral density (BMD) by the dual-energy X-ray absorptiometry (DXA) at days 58, 72, 86 and 100 of age. Feeding diet L caused a higher P digestibility (p = 0.008) measured from days 72 to 86 of age and at 100 days of age a higher BMC and BMD (p ≤ 0.01). Furthermore, the gilts of group L deposited more minerals in the body than control pigs (by 2.4 g/d, p = 0.008). It was found that BMD and BMC were positively correlated with body lean mass and digestible P intake. The results indicated that, even for very young pigs, the addition of microbial phytase instead of inorganic P increases the amount of digestible P covering the requirements of piglets for proper bone mineralisation. Furthermore, it was proved that the DXA method can be successfully applied to measure body fat and lean mass contents as well as bone mineralisation of growing pigs using the same animals.     

Digestibility and utilisation of diets composed of wet distillers' solids or soyabean meal and supplemented with liquid lysine product for growing pigs

A digestibility and balance trial was carried out to study the nutrient digestibility and utilisation of protein and energy in wet distillers' solids derived from barley or soyabean meal. Eight growing pigs (30–72 kg liveweight) were used in an 8 × 6 cyclic change-over experimental design, in which eight experimental diets were arranged 2 × 2 × 2 factorially. The corresponding factors were the protein source (wet distillers' solids (DS) or soyabean meal (SBM)), protein supply (130 or 162 g crude protein (CP) kg⁻¹ dry matter (DM)) and liquid lysine product supplementation.DS and SBM contained 565 g and 485 g CP kg⁻¹ DM, respectively, and the respective lysine contents in CP were 39 g and 64 g per 160 g N. The liquid lysine product contained 527 g CP kg⁻¹ DM and lysine in CP 193 g per 160 g N.No differences were found in the total tract digestibility of the nutrients or energy among diets composed of DS or SBM without lysine supplementation. Those diets with liquid lysine product supplementation, however, had opposite effects on the digestibility of the diets composed of the different protein sources. Lysine supplementation improved the digestibility of ash (P < 0.001), ether extract (P < 0.05) and crude carbohydrates (CCH) (P < 0.05) in diets composed of DS and adversely impaired the digestibility of organic matter and CCH (P < 0.05) in diets composed of SBM. The calculated digestibility of CP and gross energy were respectively 91.2% and 88.3% in SBM and 90.2% and 85.0% in DS. The digestible and calculated net energy contents were respectively 18.16 MJ kg⁻¹ DM and 10.73 MJ kg⁻¹ DM for SBM and 19.31 MJ kg⁻¹ DM and 10.40 MJ kg⁻¹ DM for DS.The pigs on the diets composed of DS had higher total (P < 0.001) and urea (P < 0.01) nitrogen (N) excretion in urine and lower daily retention of N (P < 0.001) than the pigs on the diets composed of SBM. The liquid lysine product supplementation of the diets decreased the total and urea N excretion in urine (P < 0.001) and improved the daily N retention (P < 0.001). With lysine supplementation, the protein utilisation of the diets composed of DS was improved to the level of the diets composed of SBM. No differences were observed in the utilisation of energy among the diets composed of different protein sources.It is concluded that DS is highly digestible, but its protein is efficiently utilised only with lysine supplementation.     

Osmotic stress enhances the competence of Beta vulgaris vacuoles to respond to inositol 1,4,5-trisphosphate

Isolated vacuoles from Beta vulgaris storage roots respond to the intracellular signalling molecule inositol 1,4,5-trisphosphate (InsP₃). Whole vacuole patch clamp enables measurement of an inward current (cytosol-directed) induced by cytosolic InsP₃ which is fully reversible upon removal of InsP₃. The reversal potentials of the InsP₃-induced whole vacuolar currents indicate a permeability ratio (P,Ca:P,K) of 200:1. Competence of vacuoles to respond to InsP₃ is dependent upon the root tissue undergoing hyperosmotic stress before vacuole isolation. The magnitude of the hyperosmotic stress and the density of InsP₃-induced current per unit membrane area are exponentially related. A standing osmotic gradient across the vacuolar membrane further enhances the InsP₃-induced current, the current being larger when there is net water flux from the cytosol to the vacuolar lumen. InsP₃-induced currents are not affected by the cytosolic free Ca²⁺ concentration. The conductance of InsP₃-induced single channel currents varied greatly between individual outside-out patches, but all showed a non-linear increase in single channel current at physiological potentials. The reversal potentials of these currents indicated a P_Ca:P_K of between 100:1 and 800:1. The significance of these findings is discussed in relation to technical aspects of monitoring InsP₃-induced currents in plant vacuoles and in the context of the physiological roles of InsP₃ and its receptor in cell water relations.