The effect of bone and analytical methods on the assessment of bone mineralization response to dietary phosphorus, phytase, and vitamin D in nursery pigs.

A total of 360 pigs (DNA 600 × 241, DNA; initially 11.9 ±â€…0.56 kg) were used in a 28-d trial to evaluate the effects of different bones and analytical methods on the assessment of bone mineralization response to dietary P, vitamin D, and phytase in nursery pigs. Pens of pigs (six pigs per pen) were randomized to six dietary treatments in a randomized complete block design with 10 pens per treatment. Dietary treatments were designed to create differences in bone mineralization and included: (1) 0.19% standardized total tract digestibility (STTD) P (deficient), (2) 0.33% STTD P (NRC [2012] requirement) using monocalcium phosphate, (3) 0.33% STTD P including 0.14% release from phytase (Ronozyme HiPhos 2700, DSM Nutritional Products, Parsippany, NJ), (4) 0.44% STTD P using monocalcium phosphate, phytase, and no vitamin D, (5) diet 4 with vitamin D (1,653 IU/kg), and (6) diet 5 with an additional 50 µg/kg of 25(OH)D3 (HyD, DSM Nutritional Products, Parsippany, NJ) estimated to provide an additional 2,000 IU/kg of vitamin D3. After 28 d on feed, eight pigs per treatment were euthanized for bone (metacarpal, 2nd rib, 10th rib, and fibula), blood, and urine analysis. The response to treatment for bone density and ash was dependent upon the bone analyzed (treatment × bone interaction for bone density, P = 0.044; non-defatted bone ash, P = 0.060; defatted bone ash, P = 0.068). Thus, the response related to dietary treatment differed depending on which bone (metacarpal, fibula, 2nd rib, or 10th rib) was measured. Pigs fed 0.19% STTD P had decreased (P < 0.05) bone density and ash (non-defatted and defatted) for all bones compared to 0.44% STTD P, with 0.33% STTD P generally intermediate or similar to 0.44% STTD P. Pigs fed 0.44% STTD P with no vitamin D had greater (P < 0.05) non-defatted fibula ash compared to all treatments other than 0.44% STTD P with added 25(OH)D3. Pigs fed diets with 0.44% STTD P had greater (P < 0.05) defatted second rib ash compared to pigs fed 0.19% STTD P or 0.33% STTD P with no phytase. In summary, bone density and ash responses varied depending on bone analyzed. Differences in bone density and ash in response to P and vitamin D were most apparent with fibulas and second ribs. There were apparent differences in the bone ash percentage between defatted and non-defatted bone. However, differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for the detection of lesions.

Lameness is defined as impaired movement or deviation from normal gait. There are many factors that can contribute to lameness, including but not limited to: infectious disease, genetic and conformational anomaly, and toxicity that affects the bone, muscle, and nervous systems. Metabolic bone disease is another cause of lameness in swine production and can be caused by inappropriate levels of essential vitamins or minerals. To understand and evaluate bone mineralization, it is important to understand the differences in diagnostic results between different bones and analytical techniques. Historically, percentage bone ash has been used as one of the procedures to assess metabolic bone disease as it measures the level of bone mineralization; however, procedures and results vary depending on the methodology and type of bone measured. Differences in bone density and ash in response to dietary P and vitamin D were most apparent in the fibulas and second ribs. There were apparent differences in the percentage of bone ash between defatted and non-defatted bone; however, the differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for detection of lesions associated with metabolic bone disease.

Requirement of digestible calcium at different dietary concentrations of digestible phosphorus for broiler chickens 3. Broiler finishers (d 25 to 35 post-hatch).

An experiment was conducted to determine the digestible calcium (Ca) and digestible phosphorous (P) requirements of 25 to 35-day-old broiler chickens. Fifteen corn-soybean meal-based diets containing 2.0, 2.5, 3.0, 3.5, and 4.0 g/kg standardized ileal digestible (SID) Ca and 2.5, 3.5, and 4.5 g/kg SID P were fed to broilers from d 25 to 35 post-hatch. Each experimental diet was randomly allocated to 6 replicate cages (8 birds per cage). Body weight and feed intake were recorded, and the feed conversion ratio was calculated. On d 35, birds were euthanized to collect the ileal digesta, tibia, and carcass for the determination of ileal Ca, and P digestibility, concentrations of ash, Ca, and P in tibia and the retention of Ca and P in the carcass. Titanium dioxide (5.0 g/kg) was included in all diets as an indigestible indicator for the ileal digestibility measurement. Feed intake and total excreta output were measured during the last 4 d of the experimental period for the measurement of apparent total tract retention of Ca and P. Fixed effects of the experiment were dietary concentrations of SID Ca and SID P and their interaction. If the interaction or main effects were significant (P < 0.05), the parameter estimates for second-order response surface model (RSM) were determined using General Linear Model procedure of SAS. The maximum response was not predicted for most of the parameters (including growth performance and tibia) as the Ca effect was linear which indicated that the highest level of Ca employed in the study may have not been high enough. The requirement of dietary SID Ca for maximization of these parameters, therefore, depends on the dietary SID P concentration when the dietary SID Ca is within 2.0 to 4.0 g/kg. However, based on the factorial analysis, the highest weight gain was observed at 3.5 g/kg SID P and 3.5 g/kg SID Ca concentrations. Tibia ash was higher in birds fed 4.5 g/kg SID P and was unaffected by dietary SID Ca concentrations. However, based on overall findings, a combination of 3.5 g/kg SID P and 3.0-3.5 g/kg SID Ca may be recommended for the optimum tibia ash. The recommended SID Ca requirements (at 3.5 g/kg SID P) for weight gain (3.5 g/kg or 6.4 g/kg total Ca) and tibia ash (3.0-3.5 g/kg or 5.5-6.4 g/kg total Ca) are lower than the current Ca recommendations (7.8 g/kg total Ca equivalent to 4.25 g/kg SID Ca; Ross, 2019) for broiler finishers, suggesting possible excess of Ca in diets formulated based on the current recommendation.

Dietary phosphorus supplementation in the diet of Pacific white shrimp (Litopenaeus vannamei) alleviated the adverse impacts caused by high Clostridium autoethanogenum protein.

The study evaluated the effects of dietary phosphorus supplementation on the fishmeal replacement with Clostridium autoethanogenum protein (CAP) in the diet of L. vannamei. Four isonitrogenous and isolipid diets were formulated: the PC diet contains 25% fishmeal, the NC, P1 and P2 diets were replaced 40% fishmeal with CAP and supplemented with 0, 0.8 and 1.6% NaH2PO4 respectively (equivalent to dietary phosphorus level of 0.96%, 1.12% and 1.27%). Sampling and V. parahaemolyticus challenge test were conducted after 50-day-feeding (initial shrimp weight 1.79 ± 0.02 g). The results showed that there were no significant differences in the growth performance of shrimp among the 4 groups. The expressions of dorsal in the gut were significantly lower in shrimp fed the P1 and P2 diets than shrimp fed the NC diet and the expression of peroxinectin in the gut was lower in shrimp fed the NC diet than others. The cumulative mortality of shrimp after V. parahaemolyticus challenge was significantly lower in shrimp fed the P2 diet than those fed the NC diet. After the challenge, genes expressions related to the prophenoloxidase activating system (proPO, lgbp, ppaf) were inhibited in the hepatopancreas of shrimp fed NC diet but activated in shrimp fed the P1 diet compared to those fed the PC diet. The AKP and T-AOC activities were higher in shrimp fed the P2 diet than those fed the other diets. The thickness of muscle layer of shrimp fed the P1 diet was thicker than that in the other groups, and significant stress damage happened in the midgut of the shrimp fed the NC diet. The abundance of Pseudoalteromonas, Haloferula and Ruegeria in shrimp fed the P1 diet was higher than those fed the other diets, while Vibrio in shrimp fed the P2 diet was higher than those fed the other diets. This indicated that a low fishmeal diet with dietary phosphorus level of 1.12% could improve the histology, enhance immune response, and increase the abundance of beneficial bacteria in the gut of shrimp. The low fishmeal diet with dietary phosphorus level of 1.27% could improve disease resistance and antioxidant capacity, but there was a possibility of damage to the gut histology as well as increasing abundance of Vibrio in the gut microbiota of shrimp.

Effects of phytase supplementation of high-plant-protein diets on growth, phosphorus utilization, antioxidant, and digestion in red swamp crayfish (Procambarus clarkii).

Fish meal is increasingly being replaced by plant protein raw materials, meanwhile, it brings phytic acid, which combines with phosphorus to form phytate phosphorus and leads to a low utilization rate of phosphorus in shrimp. To solve this problem, this study investigated the effects of phytase supplementation on growth performance, phosphorus utilization, antioxidants, and digestion in red swamp crayfish (Procambarus clarkii). Crayfish (initial mean weight: 8.69 ± 0.15 g, N = 324) were randomly divided into six groups each with three replicates of 18 individuals each, and hand-fed for 8 weeks with one of six experimental diets (50 and 490 g kg-1 animal and plant protein raw material, respectively): negative control (NC; 11.0 g kg-1 phosphorus), positive control (PC; 15 g kg-1 NaH2PO4 added to NC; 14.7 g kg-1 phosphorus), and phytase supplementation diets (P1-P4: 0.1, 0.2, 0.4, and 0.6 g kg-1 phytase added to NC, respectively). The feeding trial was performed in a micro-flow water culture system. P2 showed a significantly higher weight gain rate (WGR), specific growth rate, protein efficiency ratio, and protein retention efficiency (PRE) but showed the lowest feed conversion ratio (FCR) than other groups. Broken-line regression analyses using WGR, FCR, and PRE as evaluation indices showed that the optimal dietary phytase supplementation level was 0.233, 0.244, and 0.303 g kg-1, respectively. P2 showed the highest crude protein content of whole crayfish and abdominal muscle, and phosphorus deposition rate, which was significantly higher than that in NC and PC. P3 showed the highest calcium and phosphorus contents in whole crayfish and phosphorus content in abdominal muscle, and calcium and inorganic phosphorus content in serum, which were significantly higher than those in NC. P3 showed significantly lowest serum alkaline phosphatase, alanine aminotransferase, aspartate transaminase activities, malondialdehyde content in hepatopancreas, and highest catalase activity, which were significantly lower and higher, respectively, than those in NC and PC. In summary, the addition of 0.2-0.4 g kg-1 phytase significantly improves the growth performance, feed utilization, digestive enzyme activity, and antioxidant of P. clarkii, which has a similar effect to the direct addition of NaH2PO4 at 15 g kg-1 to the feed.

Effect of limestone solubility on mineral digestibility and bone ash in nursery pigs fed diets containing graded level of inorganic phosphorus or increasing dose of a novel consensus bacterial 6-phytase variant.

The effect of a novel consensus bacterial 6-phytase variant (PhyG) on total tract digestibility (ATTD) of minerals and bone ash was evaluated in pigs fed diets containing medium- and high-solubility limestone (MSL and HSL, 69.6 and 91.7% solubility, respectively, at 5 min, pH 3.0) in a randomized complete block design. For each limestone, eight diets were formulated: an inorganic phosphate-free negative control (NC) based on wheat, corn, soybean-meal, canola-meal and rice-barn [0.18% standardized total tract digestible (STTD) P and 0.59% Ca]; the NC supplemented with 250, 500, 1,000, or 2,000 FTU/kg of PhyG, and; the NC with added monocalcium phosphate (MCP) and limestone to produce three positive controls (0.33, 0.27, and 0.21% STTD P, and 0.75, 0.70, and 0.64% Ca, respectively; PC1, PC2, PC3). In total, 128 pigs (12.8 ± 1.33 kg, 8 pigs/treatment, housed individually) were adapted for 16 d followed by 4 d of fecal collection. Femurs were collected from euthanized pigs on day 21. Data were analyzed by one-way ANOVA with means separation by Tukey's test, and by factorial analysis (2 x 4: 2 levels of limestone solubility, 4 STTD P levels, and 2 × 5: 2 levels of limestone solubility, 5 PhyG dose levels). Phytase dose-response was analyzed by curve fitting. A consistent negative effect of HSL on ATTD P and Ca was observed in control diets (P < 0.001). Across phytase-supplemented diets, HSL reduced (P < 0.05) ATTD Ca and P (% and g/kg) compared with MSL. Across limestones, increasing phytase dose level increased (P < 0.05) ATTD P exponentially. Limestone solubility had no effect on bone ash, but PhyG linearly increased (P < 0.05) bone ash; 500 FTU/kg or higher maintained bone ash (g/femur) equivalent to PC1. In conclusion, ATTD P and Ca were reduced by a high compared with a medium soluble limestone, but the novel phytase improved ATTD P and Ca independent of limestone solubility.

Microbial phytase is added to commercial pig diets to increase phosphorus (P) availability and reduce P excretion. It is known that an excess of calcium (Ca), mostly sourced from limestone, can affect phytase efficacy. However, less is known about the impact of limestone quality. This study investigated the effect of a medium- compared to a high-soluble limestone (MSL and HSL, respectively), in combination with increasing dose levels of a novel phytase (PhyG), on mineral digestibility and bone mineralization in young pigs. Without phytase, total tract digestibility of P was lower with HSL than MSL, indicating a negative effect of more soluble limestone on mineral digestibility. Increasing the phytase dose increased digestibility of P with either limestone, and reduced the negative effect of HSL at high dose. Bone mineralization was unaffected by limestone but markedly increased by phytase. At 1,000 FTU/kg, PhyG released an estimated 1.89 or 2.32 g/kg of digestible P from monocalcium phosphate in diets containing MSL and HSL, respectively based on bone ash content. The results demonstrate the efficacy of PhyG in young pig diets whilst indicating that limestone solubility can affect phytase efficacy.

Effects of dietary phosphates from organic and inorganic sources on parameters of phosphorus homeostasis in healthy adult dogs.

BACKGROUND: The impact of dietary phosphorus (P) excess, especially on renal and cardiovascular health, has been investigated in several species, but little is known in dogs. OBJECTIVE: The aim of this study was to examine effects of different P sources on concentration and postprandial kinetics of selected parameters of P homeostasis in dogs. METHODS: Eight beagles received one control diet (P 0.5% dry matter [DM]) and three high P diets (poultry meal, NaH2PO4, and KH2PO4; P 1.7% DM) for 18d. Urine samples were collected pre- and postprandially while faeces were collected quantitatively for 5d and analysed for minerals. On day 18, blood was sampled 1h pre- and 0.5, 1, 1.5, 2, 3, 5 and 7h postprandially. RESULTS: Pi (KH2PO4, NaH2PO4) but not organic P caused an increased apparent P digestibility and significantly influenced kinetics of serum FGF23, parathyroid hormone, P, CrossLaps and bonespecific alkaline phosphatase, demonstrating a disrupted calcium (Ca) and P homeostasis with potential harm for renal, cardiovascular and skeletal health. CONCLUSIONS: Results of feeding Pi to dogs indicate distinct disturbances of Ca and P metabolism, in contrast to organic sources. The use of Pi in food can therefore not be considered as safe. Further research, especially on dose and long-term effects, is warranted.

Effects of dietary calcium and available phosphorus levels and phytase supplementation on performance, bone mineral density, and serum biochemical bone markers in aged white egg-laying hens.

Exogenous phytase supplementation increases P and Ca availability to allow for the dietary reductions without negative consequences on productivity or skeletal health. Effects of a Buttiauxella sp. phytase (BSP) supplemented in available P (avP)-reduced and Ca-reduced diets on performance, BW, eggshell quality, serum biochemical bone markers, and bone densitometry were evaluated in egg-laying hens from 68 to 78 wk of age. One hundred hens were fed 1 of 5 diets (n = 20/treatment), including a positive control (PC) with 0.35% avP and 3.5% Ca, and the PC moderately reduced in avP and Ca levels by 0.187 and 0.159% of the diet (by 53 and 4.5%), respectively, (NC1) or severely reduced by 0.231 and 0.275% of the diet (by 66 and 7.9%), respectively, (NC2). Other diets were the NC1 or NC2 supplemented with BSP at 600 FTU/kg (NC1 + BSP or NC2 + BSP, respectively). Egg production and feed conversion ratio were maintained by NC1 but were 11.9% lower and 12.3% higher, respectively, with the NC2 than the PC, which was alleviated by supplemental BSP. Diet effects on FI and eggshell quality followed a similar pattern. Body weight was 2.9% lower for NC1, and 6.1% for NC2 than the PC; BSP alleviated the decreased BW. Serum pyridinoline (bone resorption marker) was 20 to 27% higher in NC2 hens than in the other groups, with no effects on other bone markers. Total and trabecular space bone mineral density in the proximal metaphysis were 8.4 and 15.2% lower for NC1, respectively, and 12.1 and 26.7% lower for NC2, respectively, than PC. Supplemental BSP completely alleviated the decreased bone densitometry measures in NC1, but only partially in NC2. The NC1 hens maintained performance but had decreased BW and bone quality; phytase supplementation restored productivity, BW, and bone quality. The Ca and avP deficiencies in the NC2 hens relative to other groups were partially alleviated by the 600 FTU/kg BSP.

Additivity of apparent and standardized ileal digestibility of phosphorus in mixed diets containing corn and soybean meal fed to broiler chickens.

Phosphorus (P) is an integral part of diet formulation for broiler chickens as P is required for various biochemical processes essential to life. A study was designed to examine the additivity of apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of P in mixed diets containing corn and soybean meal (SBM) with or without phytase supplementation. Birds were fed a commercial starter diet from day 0 to 21 after hatching and then allotted to 7 dietary treatments in a randomized complete block design with the BW as a blocking factor. Four semipurified diets were prepared to contain corn or SBM as the sole source of P with or without the addition of phytase at 1,000 phytase units/kg of diet. Two mixed diets were also prepared to contain corn and SBM with or without the addition of phytase at 1,000 phytase units/kg diet. A P-free diet (PFD) was formulated to determine the basal ileal endogenous loss of P. There were 16 replicate cages of the PFD and 8 replicate cages of the 6 experimental diets, with 8 birds per replicate cage for a total of 512 birds. Diets were fed for 3 d. The ileal digesta of birds were collected from the distal two-thirds of the ileum on day 24 after hatching. The SID of P in corn and SBM were 52.2 and 65.4%, respectively (SEM = 1.37). The addition of phytase improved (P < 0.05) both the AID and SID of P in the corn, SBM, and mixed diets. The determined AID or SID in the corn and SBM with or without phytase was used to predict the AID or SID in the mixed diets. There were no differences between the predicted and determined digestibility values in the mixed diets for either AID or SID of P and thus additive. Phytase supplementation of the mixed diet did not influence the additivity of AID or SID. In conclusion, the AID or SID of P in the corn and SBM was additive in the mixed diets containing corn and SBM with or without the addition of phytase.

More Bone with Less Minerals? The Effects of Dietary Phosphorus on the Post-Cranial Skeleton in Zebrafish.

Dietary phosphorus (P) is essential for bone mineralisation in vertebrates. P deficiency can cause growth retardation, osteomalacia and bone deformities, both in teleosts and in mammals. Conversely, excess P supply can trigger soft tissue calcification and bone hypermineralisation. This study uses a wide range of complementary techniques (X-rays, histology, TEM, synchrotron X-ray tomographic microscopy, nanoindentation) to describe in detail the effects of dietary P on the zebrafish skeleton, after two months of administering three different diets: 0.5% (low P, LP), 1.0% (regular P, RP), and 1.5% (high P, HP) total P content. LP zebrafish display growth retardation and hypomineralised bones, albeit without deformities. LP zebrafish increase production of non-mineralised bone matrix, and osteoblasts have enlarged endoplasmic reticulum cisternae, indicative for increased collagen synthesis. The HP diet promotes growth, high mineralisation, and stiffness but causes vertebral centra fusions. Structure and arrangement of bone matrix collagen fibres are not influenced by dietary P in all three groups. In conclusion, low dietary P content stimulates the formation of non-mineralised bone without inducing malformations. This indicates that bone formation and mineralisation are uncoupled. In contrast, high dietary P content promotes mineralisation and vertebral body fusions. This new zebrafish model is a useful tool to understand the mechanisms underlying osteomalacia and abnormal mineralisation, due to underlying variations in dietary P levels.

How do pigs deal with dietary phosphorus deficiency?

Feeding strategies for growing monogastric livestock (particularly pigs) must focus on maximising animal performance, while attempting to reduce environmental P load. Achieving these goals requires a comprehensive understanding of how different P feeding strategies affect animal responses and an ability to predict P retention. Although along with Ca, P is the most researched macromineral in pig nutrition, knowledge gaps still exist in relation to: (1) the effects of P feed content on feed intake (FI); (2) the impact of P intake on body composition; (3) the distribution of absorbed P to pools within the body. Here, we address these knowledge gaps by gathering empirical evidence on the effects of P-deficient feeds and by developing a predictive, mechanistic model of P utilisation and retention incorporating this evidence. Based on our statistical analyses of published literature data, we found: (1) no change in FI response in pigs given lower P feed contents; (2) the body ash-protein relationship to be dependent upon feed composition, with the isometric relationship only holding for pigs given balanced feeds and (3) the priority to be given towards P retention in soft tissue over P retention in bones. Subsequent results of the mechanistic model of P retention indicated that a potential reduction in P feeding recommendations could be possible without compromising average daily gain; however, such a reduction would impact P deposition in bones. Our study enhances our current knowledge of P utilisation and by extension excretion and could contribute towards developing more accurate P feeding guidelines.

Dietary Phosphate and the Forgotten Kidney Patient: A Critical Need for FDA Regulatory Action.

Careful dietary management that reduces high phosphate intake is recommended to slow the progression of chronic kidney disease (CKD) and prevent complications of CKD and may help reduce chronic disease risks such as incident CKD associated with high phosphate intake in the healthy general population. For patients treated with maintenance dialysis, control of serum phosphorus levels is considered a marker of good care and requires a coordinated plan that limits dietary phosphate intake, uses oral phosphate binders, and provides an adequate dialysis prescription. Even with traditional thrice-weekly hemodialysis or peritoneal dialysis, use of phosphate binders, and a concerted effort to limit dietary phosphate intake, adequately controlled serum phosphorus levels are not possible in all dialysis patients. Efforts to limit phosphate intake are thwarted by the underestimated and unquantified phosphate content of processed foods and some medications due to the hidden presence of phosphate additives or excipients added during processing or drug formulation. Effectively limiting phosphate intake could potentially be achieved through simple US Food and Drug Administration regulatory actions. Mandatory labeling of phosphate content on all packaged foods and drugs would enable identification of healthy low-phosphate foods and medications and permit critically important control of total phosphate intake. Simple changes in regulatory policy and labeling are warranted and would enable better management of dietary intake of phosphate at all stages of kidney disease, as well as potentially reduced health risks in the general population.

Phosphorus ingestion with a high-carbohydrate meal increased the postprandial energy expenditure of obese and lean individuals.

OBJECTIVES: Phosphorus ingestion with glucose was reported to stimulate the postprandial peripheral uptake of both phosphorus and glucose, a process that favors energy production. The aim of this study was to determine whether phosphorus ingestion with a meal can affect energy metabolism. METHODS: Overnight fasted men (eight lean and seven obese) consumed a high-carbohydrate meal (648 kcal) with either placebo or phosphorus (500 mg) tablets in a random order. Energy expenditure and substrate oxidation were monitored for 240 min using ventilated hood indirect calorimetry. RESULTS: Phosphorus ingestion with a meal increased the postprandial energy expenditure of both lean and obese individuals (P < 0.001), although in different patterns. Alterations in postprandial substrate oxidation was highly noticeable from time 120 min onward, where phosphorus-treated lean participants exhibited a significant decrease in respiratory quotient. CONCLUSION: Phosphorus ingestion with a high-carbohydrate meal alters postprandial energy metabolism mainly by enhancing postprandial energy expenditure that may ultimatly favor weight loss.

Effect of phytase enzyme on growth performance, serum biochemical alteration, immune response and gene expression in Nile tilapia.

The present study aimed to investigate the effect of low phosphorus diet with or without different levels of phytase enzyme supplementation on growth performance, body composition, nutrient retention efficiency, gene expression, and health status of A. hydrophila challenged fish. A total of 240 monosex males of Nile tilapia (Oreochromis niloticus) with an average body weight of 23.19 ±â€¯0.15 g/fish were used. Fish were randomly chosen and divided into 4 equal groups (60 fish per group), with 3 subgroups containing 20 fish as a replicate. Group 1, was fed on a diet containing 100% P, group 2, was fed on a diet containing 50% P, group 3 and 4, were fed on low P with 500 or 1000 units of phytase/Kg respectively. It was observed that the 50% phosphorus diet significantly reduced body weight, feed intake, feed conversion ratio (FCR), and protein efficiency ratio (PER) compared to Nile tilapia fish fed on the diet containing 100% phosphorus. In contrast, fish fed on the diet containing 50% phosphorus supplemented by 500 or 1000 phytase units/kg significantly (P ≤ 0.05) increased final body weight (FBW), total body gain (TBG), average daily gain (ADG), and weight gain compared to Nile tilapia fed on the same diet or fed on the diet containing normal phosphorus without phytase supplementation. Different phosphorus and phytase supplementation levels had no significant effect on serum total protein, albumin, and globulin concentrations, meanwhile, phytase supplementation increased serum calcium and phosphorus levels. Nile tilapia fed on phytase supplementation had an increase in body protein, lipid content, and nutrient utilization efficiency compared to Nile tilapia fed on the diet containing 100% phosphorus. Nile tilapia fed on low dietary phosphorus showed an increase in mortality after infection and a decrease in phagocytosis and neutrophil compared to fish fed on normal phosphorus. Phytase supplementation, made immune response parameters return to its normal values and the pathological lesions of liver, spleen, stomach, and intestine were reduced. Moreover, normal phosphorus significantly up-regulated lipoprotein lipase (LPL) mRNA expression and down-regulated fatty acid synthase (FAS) mRNA in Nile tilapia's liver while low phosphorus with or without phytase supplementation reduced LPL expression and relatively up-regulated FAS.

Effects of dietary calcium to phosphorus ratio and addition of phytase on growth performance of nursery pigs.

Two studies were conducted to evaluate the growth performance and percentage bone ash of nursery pigs fed various combinations of Ca and P provided by inorganic sources or phytase. In Exp. 1, pens of pigs (n = 720, initially 6.1 ± 0.98 kg) were blocked by initial BW. Within blocks, pens were randomly assigned to one of six treatments (12 pens per treatment) in a three-phase diet regimen. Treatments were arranged in a 2 × 3 factorial with main effects of Ca (0.58% vs. 1.03%) and standardized total tract digestible (STTD) P (0.33% and 0.45% without phytase, and 0.45% with 0.12% of the P released by phytase). During treatment period, Ca × P interactions were observed for all growth criteria (P < 0.05). When diets had low Ca, pigs fed 0.45% STTD P with phytase had greater (P < 0.01) ADG and ADFI than those fed 0.33% or 0.45% STTD P without phytase. When high Ca was fed, ADG and ADFI were similar among pigs fed 0.45% STTD P with or without phytase and were greater than those fed 0.33% STTD P. Gain:feed was reduced (P < 0.01) when high Ca and low STTD P were fed relative to other treatments. On d 21, radiuses were collected from 1 pig per pen for bone ash analysis. Pigs fed 0.33% STTD P had decreased (P < 0.05) percentage bone ash than those fed 0.45% STTD P with or without phytase when high Ca was fed, but this P effect was not observed for low Ca diets (Ca × P interaction, P = 0.007). In Exp. 2, 36 pens (10 pigs per pen, initially 6.0 ± 1.08 kg) were used in a completely randomized design. Treatments were arranged in a 2 × 3 factorial with the main effects of STTD P (at or above NRC [NRC. 2012. Nutrient Requirements of Swine. 11th rev. ed. Washington (DC): National Academic Press.] requirement estimates) and total Ca (0.65, 0.90, and 1.20%). Experimental diets were fed during phases 1 and 2, followed by a common phase 3 diet. Diets at NRC (2012) P level contained 0.45% and 0.40% STTD P, compared with 0.56% and 0.52% for diets greater than the NRC (2012) estimates, in phase 1 and 2, respectively. During treatment period, increasing Ca decreased (linear, P = 0.006) ADG, but increasing STTD P marginally increased (P = 0.084) ADG, with no Ca × P interaction. When diets contained NRC (2012) P levels, pigs fed 1.20% Ca had decreased (P < 0.05) G:F than those fed 0.65% or 0.90% Ca; however, when high STTD P were fed, G:F was not affected by Ca (Ca × P interaction, P = 0.018). In conclusion, excess Ca decreased pig growth and percentage bone ash when diets were at or below NRC (2012) requirement for STTD P, but these negative effects were alleviated by adding monocalcium P or phytase to the diet.

Reducing mineral usage in feedlot diets for Nellore cattle: I. Impacts of calcium, phosphorus, copper, manganese, and zinc contents on microbial efficiency and ruminal, intestinal, and total digestibility of dietary constituents.

This study evaluated intake, microbial efficiency, and ruminal, small and large intestinal, and total digestibility of DM, OM, CP, and NDF, as well as availability of Ca, P, Mg, Na, K, Cu, Mn, and Zn in Zebu cattle fed with or without supplemental sources of Ca and P or a micromineral premix. Five rumen- and ileum-cannulated Nellore bulls (BW = 200 ± 10.5 kg; 9 mo) were used in the experiment, distributed in a 5 × 5 Latin square design. The experiment was developed in a 2 × 2 + 1 factorial design to measure the effects of mineral supplementation on intake, digestibility, and site of nutrient absorption. The factors consisted of 2 Ca and P levels (macromineral factor; CaP+ or CaP-) and 2 microminerals levels (micromineral factor; CuMnZn+ or CuMnZn-). In addition, a treatment with alimentary restriction (REST) was evaluated at 1.7% of BW. Nutrient fluxes were measured in the omasum and ileum, in addition to intake and fecal excretion. Microbial efficiency was estimated using purine derivative excretion. Dry matter, OM, NDF, CP intake, and total digestibility were not affected ( ≥ 0.058) by the absence of Ca, P, Cu, Mn, and Zn supplementation. Intake of Ca, P, and Mg were reduced ( < 0.01) by CaP-. The absence of CuMnZn reduced ( < 0.01) Cu, Mn, and Zn intake. Ruminal recycling of P, Na, and K is significant for increasing the influx of these minerals to the digestive tract; however, influences of treatments were not observed. The small and large intestines contributed to mineral absorption in different proportions ( < 0.05), according to minerals and treatments. Because of the similarity ( > 0.05) of OM, NDF, and CP digestion sites and coefficients, we assume that omitting supplemental sources of Ca, P, Cu, Mn, and Zn may be an option in raising cattle on feedlots. If supplementation is viable, knowledge about the specific absorption site of each mineral could positively impact choices about the supplemental source.

Reducing mineral usage in feedlot diets for Nellore cattle: II. Impacts of calcium, phosphorus, copper, manganese, and zinc contents on intake, performance, and liver and bone status.

Weaned Nellore bulls ( = 36; 274 ± 34 kg) were used in a randomized block design with a 2 × 2 factorial arrangement of treatments to evaluate intake, fecal excretion, and performance with different concentrations of minerals. Experimental diets were formulated with 2 concentrations of Ca and P (macromineral factor; diet supplying 100% of Ca and P according to BR-CORTE () [CaP+] or diet without limestone and dicalcium phosphate [CaP-]) and 2 concentrations of microminerals (micromineral factor; diet with supplementation of microminerals [Zn, Mn, and Cu; CuMnZn+] or diet without supplementation of microminerals [Zn, Mn, and Cu; CuMnZn-]). The factor CaP- was formulated without the addition of limestone and dicalcium phosphate, and the factor CuMnZn- was formulated without inorganic supplementation of microminerals (premix). The diets were isonitrogenous (13.3% CP). Intake was individually monitored every day. Indigestible NDF was used as an internal marker for digestibility estimates. The bulls were slaughtered (84 or 147 d on feed), and then carcass characteristics were measured and liver and rib samples were collected. Feed, feces, rib bones, and liver samples were analyzed for DM, ash, CP, ether extract (EE), Ca, P, Zn, Mn, and Cu. There were no significant interactions ( ≥ 0.06) between macro- and micromineral supplementation for any variables in the study. Calcium, P, and micromineral concentrations did not affect ( ≥ 0.20) intake of DM, OM, NDF, EE, CP, TDN, and nonfiber carbohydrates (NFC). Calcium and P intake were affected ( < 0.01) by macromineral factor. Animals fed without Ca and P supplementation consumed less of these minerals. Dry matter and nutrient fecal excretion (OM, NDF, EE, CP, and NFC) were similar ( ≥ 0.23) among all factors. Performance and carcass characteristics were similar ( ≥ 0.09) among diets. The content of ash in rib bones was not affected by diets ( ≥ 0.06). Plasma P and phosphatase alkaline concentrations were similar ( ≥ 0.52) among diets. Supplementation of microminerals decreased ( < 0.01) plasma Ca concentration; nevertheless, all analyzed blood metabolites were within the reference values. Supplementation of Ca and P increased ( < 0.01) fecal excretion of these minerals. These results indicate that mineral supplementation (Ca, P, Zn, Mn, and Cu) of conventional feedlot diets for Nellore bulls may be not necessary. Dietary reductions in these minerals would represent a decrease in the cost of feedlot diets. Dietary reduction in Ca and P content cause a decrease in fecal excretion of these minerals, which, in turn, represents an opportunity to reduce the environmental impact of feedlot operations.

Response of Npt2a knockout mice to dietary calcium and phosphorus.

Mutations in the renal sodium-dependent phosphate co-transporters NPT2a and NPT2c have been reported in patients with renal stone disease and nephrocalcinosis, but the relative contribution of genotype, dietary calcium and phosphate to the formation of renal mineral deposits is unclear. We previously reported that renal calcium phosphate deposits persist and/or reappear in older Npt2a-/- mice supplemented with phosphate despite resolution of hypercalciuria while no deposits are seen in wild-type (WT) mice on the same diet. Addition of calcium to their diets further increased calcium phosphate deposits in Npt2a-/-, but not WT mice. The response of PTH to dietary phosphate of Npt2a-/- was blunted when compared to WT mice and the response of the urinary calcium x phosphorus product to the addition of calcium and phosphate to the diet of Npt2a-/- was increased. These finding suggests that Npt2a-/- mice respond differently to dietary phosphate when compared to WT mice. Further evaluation in the Npt2a-/- cohort on different diets suggests that urinary calcium excretion, plasma phosphate and FGF23 levels appear to be positively correlated to renal mineral deposit formation while urine phosphate levels and the urine anion gap, an indirect measure of ammonia excretion, appear to be inversely correlated. Our observations in Npt2a-/- mice, if confirmed in humans, may be relevant for the optimization of existing and the development of novel therapies to prevent nephrolithiasis and nephrocalcinosis in human carriers of NPT2a and NPT2c mutations.

Standardized total tract digestibility of phosphorus in flaxseed meal fed to growing and finishing pigs without or with phytase supplementation.

Two experiments were conducted to determine the apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of P in flaxseed meal (FM) and the effect of dietary microbial phytase on the digestibility of P in FM fed to growing and finishing pigs. In Exp. 1, eighteen growing barrows (26.6 ± 1.8 kg BW) were allotted to 1 of 3 experimental diets consisting of a diet containing 32% FM that was fed with or without phytase at 500 phytase units (FTU/kg and a P-free diet in a completely randomized design to give 6 replicates per diet. The experimental period lasted 12 d including first 7 d for adaptation and 5 d for total collection of feces. Pigs were fed their assigned diets at 4% of BW at the beginning of the experiment. The daily feed allowance was offered in 2 equal portions at 0800 and 1600 h. All experimental diets were provided in mash form. Results indicated that pigs fed the diets containing FM with dietary phytase had less ( < 0.05) fecal P concentration and daily P output than those fed the diets without phytase supplementation. Also, phytase supplementation increased ( < 0.05) the ATTD of P of the diets containing FM from 37.3% to 51.8% and STTD of P of the diets containing FM from 43.2% to 57.7%. The basal endogenous P losses (EPL) was calculated at 140 ± 11 mg/kg of DMI in growing pigs fed the P-free diet. In Exp. 2, eighteen finishing pigs (78.7 ± 2.4 kg BW) were randomly allotted to 1 of 3 dietary treatments. The experimental diets and procedures were similar to those described in Exp. 1. Similar to Exp. 1, pigs fed FM diets with phytase supplementation had less ( < 0.05) P concentration in feces than those fed diets without phytase supplementation. Also, daily P output was reduced ( = 0.08) when pigs were fed the FM diets with phytase compared to those fed the FM diets without phytase. The ATTD of P in FM diets was increased ( < 0.01) from 31.4% to 45.8%, whereas the STTD of P in FM diets was increased ( < 0.01) from 37.8% to 52.3% as a result of phytase supplementation. The basal EPL was calculated at 164 ± 19 mg/kg of DMI in finishing pigs fed the P-free diet. In conclusion, the ATTD and STTD of P in FM fed to growing pigs were 37.3% and 43.2%, respectively, whereas respective values for finishing pigs were 31.4%, and 37.8%, respectively. Also, dietary phytase supplementation improved both ATTD and STTD of P in FM for both stages of pigs by an average of 33%.

Effect of low levels of dietary available phosphorus on phosphorus utilization, bone mineralization, phosphorus transporter mRNA expression and performance in growing pigs.

A study was conducted to examine the effects of different dietary levels of available phosphorus (aP) on P excretion, bone mineralization, performance and the mRNA expression of sodium-dependent P transporters in growing pigs. Sixty-day old growing pigs (n = 54) with an average initial BW of 19.50 ± 1.11 kg were randomly allocated to a control diet (C) containing 0.23% available phosphorus (aP), T1 containing 0.17% aP and T2 containing 0.11% aP. There were 6 pens per treatment with 3 pigs per pen. Body weight and feed intake were measured weekly. At the end of each week, one pig from each pen was housed in a metabolic crate for 24 h to collect fecal and urine samples and then sacrificed to obtain third metacarpal (MC3) bones and jejunal and kidney samples. Bones were scanned by Dual Energy X-ray Absorptiometry (DEXA). Fecal and urine samples were sub-sampled and analyzed for P content. The expression of P transporter mRNA in jejunum and kidney samples was measured using quantitative real-time polymerase chain reaction (qRT-PCR). Data were analyzed using GLM procedure of the Statistical Analysis System (SAS Institute version 9.2). Pigs fed the T2 diet had reduced (P < 0.05) average daily gain (ADG) and gain to feed (G:F) compared to those fed the C diet during week 2. Overall, ADG and G:F were also reduced (P < 0.05) in pigs fed the T2 diet compared to those fed the C and T1 diets. Bone mineral density (BMD) and bone mineral content (BMC) were reduced (P < 0.05) in pigs fed the T2 diet compared to those fed the C diet throughout the experiment. At week 1, jejunal mRNA expression of Na (+)-dependent phosphate transporter 2 (SLC34A2) was increased (P < 0.01) in pigs fed the T2 diet compared to C diet. Renal mRNA expression of Na(+)-dependent phosphate transporter 1 (SLC34A1) and SLC34A3 were increased (P < 0.05) in pigs fed the T2 diet compared to those fed the C diet at week 2 and was accompanied by lower (P < 0.05) urinary P in pigs fed the T2 diet during week 2 and week 3. In conclusion, growing pigs are highly sensitive to low dietary P as shown by reduced ADG, bone mineralization and urinary P level, but moderate reduction in dietary P up to 0.17% aP in the diet has the potential to reduce environmental pollution by reducing P concentration in swine manure and without compromising performance.

Effects of dietary calcium and phosphorus on reproductive performance and markers of bone turnover in stall- or group-housed sows.

Increasing productivity and new housing standards necessitate a reevaluation of nutrient requirements for sows, including minerals. The objective of this study was to determine if the recommended levels of dietary Ca and P are adequate for sows housed in groups and that, therefore, have the potential for increased mobility. A total of 180 multiparous sows and gilts were assigned to 1 of 6 treatments. Treatments, arranged as a 3 × 2 factorial, included the main effects of dietary Ca:P-0.70:0.55% Ca:P (as-fed basis; control), 0.60:0.47% Ca:P (as-fed basis; Low CaP), and 0.81:0.63% Ca:P (as-fed basis; High CaP)-and housing-stalls or groups. The trial was initiated when sows were moved from the breeding stalls to the gestation room at wk 4 or 5 after breeding. Sows were initially fed 2.3 kg/d. This allotment was increased to 3.0 kg/d 2 wk prior to farrowing. Group-housed sows, fed in individual stalls, were allowed access to a loafing area after feeding. Serum samples were collected at the start of the trial and on d 100 of gestation, and both serum and milk samples were collected at mid lactation and prior to weaning. Neither diet nor housing had an effect on the total number of piglets born, ADG from birth to weaning, or weaning weight ( > 0.10). The number of live-born piglets and birth weight were unaffected by diet ( > 0.10) but were improved by group housing relative to stalls ( < 0.05). In late gestation, group-housed sows fed the Low CaP diet had reduced serum Ca (diet × housing interaction, = 0.02), and the greatest reduction (between d 28 and 100 of gestation) in serum P level was observed in group-housed sows fed the Low CaP diet (diet × housing interaction, = 0.04). Osteocalcin and pyridinoline, markers of bone formation and resorption, respectively, were unaffected by diet or housing ( > 0.10). Results from these studies imply that the level of dietary Ca and P recommended by the NRC is adequate for sows of modern genetics, whether housed in stalls or groups.