High Dietary Ca Research Articles

Renal expression of calcium and phosphorus transporters: contrasting responses to dietary calcium and microbial phytase in broilers and growing pigs.

Enhanced dietary Ca intake linearly increases intestinal Ca absorption in pigs, but not in broilers, suggesting potential differences in whole body Ca homeostasis. To determine the role of kidney in Ca homeostasis in these species, we varied in growing pigs in experiment (Exp) 1, the dietary Ca content 2·0 v. 9·6 g/kg and phytase 0 v. 500 FTU/kg, in broilers, in Exp 2 the dietary Ca/retainable P from 1·3 to 2·8 and phytase 0 v. 1000 FTU/kg, and in Exp 3 dietary Ca/P from 0·50 to 1·75. Increasing dietary Ca reduced renal mRNA expression of Ca-related transporters (TRPV5, TRPV6, CaBP-D28k and NCX1) and tight junctions (CLDN-12 and -16) in pigs, indicating Ca reabsorption was reduced to maintain Ca homeostasis. In broilers (Exp 2), high dietary Ca increased renal TRPV6, CaBP-D28k and CLDN-2 mRNA, indicating an increased capacity for Ca reabsorption. Moreover, the effect of dietary Ca was enhanced by inclusion of dietary phytase in pigs but reduced in broilers. Furthermore, increasing dietary Ca upregulated inorganic phosphate transporter 1 (PiT-1), while phytase downregulated xenotropic and polytropic retrovirus receptor 1 (XPR1) mRNA expression in pigs; in broilers, dietary Ca downregulated renal mRNA expression of Na-dependent phosphate transporter IIa (NaPi-IIa), PiT-1, PiT-2 and XPR1, while phytase downregulated NaPi-IIa but upregulated PiT-2 and XPR1 mRNA expression. In Exp 3, Ca/P effect on transporter mRNA expression was largely consistent with Exp 2. In conclusion of this study, together with previously measured data about Ca and P homeostasis, in pigs the kidneys play a more regulatory role in Ca homeostasis than in broilers where the intestine is more important for regulation.

Effects of feed particle size, calcium concentration and phytase supplementation on InsP6 degradation in broiler chickens fed pelleted diets

ABSTRACT 1. The objective of the trial was to study the single and interactive effects of feed particle size in pelleted feed, dietary calcium (Ca) concentration and microbial phytase supplementation in broiler chickens. The studied traits were myo-inositol (1,2,3,4,5,6) hexakis (dihydrogen phosphate) (InsP6) degradation, pre-caecal digestibility of phosphorus (P), Ca and amino acids (AA) and retention of P, Ca and nitrogen (N). 2. Male Ross 308 broiler chickens were housed in metabolism units in groups of 10 and allocated to one of eight diets with seven pen replicates per diet. The 2 × 2 × 2-factorial arrangement included coarse and fine feed particle size (309 or 222 µm), low and high Ca concentration (4.9 and 7.2 g/kg) and without or with phytase supplementation (1,000 FTU/kg). 3. Pre-caecal InsP6 disappearance was higher with coarse than fine feed particle size when no phytase was added (54 vs. 48%) but not when phytase was added (74%; p = 0.046). High dietary Ca feeds decreased pre-caecal InsP6 disappearance (67 to 59%) and P digestibility (65 to 55%; p < 0.001). Gizzard pH was lower with coarse than fine feed particle size and higher with high Ca than low Ca (p < 0.001). Pre-caecal digestibility of most AA was approximately 3.5%-points lower with high Ca without phytase compared to the other treatments (p ≤ 0.047). Coarse feed particle size caused higher pre-caecal AA digestibility than fine particle size (~2%-points; p ≤ 0.031). InsP6 disappearance in the crop increased at high Ca concentration when phytase was added (22 vs. 37%; p = 0.011). 4. Coarser feed particle size in pellets increased gastrointestinal InsP6 degradation and nutrient digestibility, likely owing to effects on the gizzard functions. Additional Ca supply exerted antinutritive effects that was not compensated for by using coarser feed particles.

Inorganic Phosphorus Supplementation in Diets for Farmed Shrimp: Performance and Digestibility of Juvenile Litopenaeus vannamei Fed Monosodium Phosphate, Monoammonium Phosphate, Magnesium Phosphate, and Monocalcium Phosphate

Inorganic phosphates have been used in shrimp feeds to meet phosphorus (P) needs, which may be quantitatively affected by the dietary content of P and other minerals and digestibility. In the present study, performance and digestibility were determined simultaneously in juvenile shrimp (Litopenaeus vannamei) during the grow‐out period (4–23 g ind wt). Shrimps were fed plant‐based diets supplemented with different phosphate ingredients: monosodium phosphate (MSP), monoammonium phosphate (MAP), magnesium phosphate (MgP), and monocalcium phosphate (MCP). The total P level in test diets was 0.79%. MSP supplementation was tested at two dietary calcium:phosphorus ratios to check for possible effects upon shrimp performance and digestibility. A negative control diet was tested with a similar formulation without inorganic phosphate supplementation (0.47% total P). Shrimp was reared in a recirculated clear water tank system (35 ppt salinity, 30°C, 100 individuals/m3) with continuous feeding provided by automatic pellet delivery. Feces were sampled 4–5 times daily throughout the trial. A negative control diet produced P limitation and significantly reduced shrimp performances compared to P‐supplemented diets (P &lt; 0.05). Performance of shrimp‐fed diets containing inorganic phosphates varied for feed conversion rate: 1.24–1.34, survival: 87%–94%, growth: 2.50–2.59 g/week, and no significant difference was found among dietary treatments (P &gt; 0.05). Inorganic P supplementation resulted in significantly higher dry matter, crude protein, ash, and P apparent digestibility coefficient (ADC) compared to P limited negative control diet (P &lt; 0.05). In addition, reduced P ADC (P &lt; 0.05) was checked with higher dietary Ca:P ratio. Phosphorus digestibility in test diets showed significant differences: MSP &gt; MCP, MgP, and MSP + CaCO3; MAP &gt; MSP + CaCO3 (P &lt; 0.05). Digestible P (%) could then be calculated in test diets: MgP (4.77 g Pdig/kg of feed), MSP + CaCO3 (4.81 g Pdig/kg of feed), MCP (5.0 g Pdig/kg of feed), MAP (5.13 g Pdig/kg of feed), and MSP (5.30 g Pdig/kg of feed). Digestible P in phosphate sources (%) could then be calculated: 88.2% MSP, 84.2% MAP, 79% MCP, 72.2% MgP, and 71.8% MSP + CaCO3. Shrimp whole body and exoskeleton phosphorus content also showed significantly higher values in P‐supplemented diets compared to control, regardless of the source tested (P &lt; 0.05). It is concluded that tested phosphates were suitable for feeding juvenile shrimp with MSP, MAP, and MgP diets showing high feed efficiency, while MSP and MAP diets resulted in numerically increased growth, coinciding with numerically higher P digestibility in these phosphates. The elevated P digestibility in the phosphates seems related to their water solubility, though MgP, less soluble and digestible in comparison to MSP and MAP, could also produce good performance.

Rabbits (Oryctolagus cuniculus) increase caecal calcium absorption at increasing dietary calcium levels.

Hindgut fermenting herbivores from different vertebrate taxa, including tortoises, and among mammals some afrotheria, perissodactyla incl. equids, several rodents as well as lagomorphs absorb more calcium (Ca) from the digesta than they require, and excrete the surplus via urine. Both proximate and ultimate causes are elusive. It was suggested that this mechanism might ensure phosphorus availability for the hindgut microbiome by removing potentially complex-building Ca from the digesta. Here we use Ussing chamber experiments to show that rabbits (Oryctolagus cuniculus) maintained on four different diets (six animals/diet) increase active Ca absorption at increasing Ca levels. This contradicts the common assumption that at higher dietary levels, where passive uptake should be more prevalent, active transport can relaxand hence supports the deliberate removal hypothesis. In the rabbits, this absorption was distinctively higher in the caecum than in the duodenum, which is unexpected in mammals. Additional quantification of the presence of two proteins involved in active Ca absorption (calbindin-D9K CB; vitamin D receptor, VDR) showed higher presence with higher dietary Ca. However, their detailed distribution across the intestinal tract and the diet groups suggests that other factors not investigated in this study must play major roles in Ca absorption in rabbits. Investigating strategies of herbivores to mitigate potential negative effects of Ca in the digesta on microbial activity and growth might represent a promising area of future research.

Negative dietary cation-anion difference and amount of calcium in prepartum diets: Effects on urine and serum minerals

Negative dietary cation-anion difference (DCAD) diets fed prepartum induce a compensated metabolic acidosis, which stimulates metabolic Ca flux before calving and decreases clinical and subclinical hypocalcemia after calving. Effects of low or high dietary Ca in these diets are unclear. Our objective was to determine the effects of inducing a prepartum metabolic acidosis and the amount of dietary Ca on urinary mineral excretion and serum mineral concentrations during the transition period in multiparous Holstein cows (n = 81). Treatment diets fed during the last 28 d before calving were (1) positive DCAD, +6 mEq/100 g of dry matter (DM), target urine pH >7.5, low dietary Ca (0.40% DM; CON); (2) negative DCAD, −24 mEq/100 g of DM, target urine pH 5.5 to 6.0, low dietary Ca (0.40% DM; ND); or (3) negative DCAD, −24 mEq/100 g of DM, target urine pH 5.5 to 6.0, high dietary Ca (2.0% DM; NDCA). Urine was sampled on −21, −14, −7, +1, +2, and +7 d relative to calving. Blood samples were collected on d −30, −21, −14, −7, −4, −2, −1, 0, 1, 2, and 4 relative to parturition. Preplanned treatment contrasts were (1) CON versus ND and NDCA, and (2) ND versus NDCA. Cows fed ND or NDCA had increased urinary volume and excretion of Ca, Cl, and SO4−2, along with decreased excretion of K. Supplementation of Ca to the acidogenic diet increased urinary excretion of Ca at d −21, but this difference lessened as cows approached parturition. Additional Ca increased prepartum urinary excretion of K and Cl. Differences in excretion decreased quickly after parturition, although urinary excretion of Cl remained greater for cows previously fed negative DCAD diets. Acidogenic diets increased serum K and Cl concentrations prepartum. Supplemental Ca decreased serum P relative to ND. During d 0 to 4, serum P and K were greater for cows fed ND or NDCA than for cows fed CON; these differences disappeared by d 4. Acidogenic diets increased serum Ca by increasing Ca flux and excretion in urine. Supplemental Ca in the acidogenic diet modestly increased Ca excretion early in the close-up period.

Short-Term Dietary Calcium Fortification Increases Fecal Saturated Fat Content and Reduces Serum Lipids in Men

The effect of dietary calcium on fecal fatty acid excretion and serum lipids was tested in a randomized, single-blind metabolic study in 13 healthy men with moderate hypercholesterolemia. A low calcium base diet containing 34% of energy from fat, 13% from saturated fatty acids, 240 mg cholesterol/d and 410 mg Ca/d was compared with a fortified version in which calcium citrate malate was added to orange juice, (550 mg) muffins (750 mg), and two tablets (500 mg) for a total calcium intake of 2200 mg/d. Fecal collections (72 h, d 8, 9, 10) and blood from fasting subjects for lipids and lipoproteins (d 9, 10, 11) were obtained. The percentage of dietary saturated fat excreted per day increased from 6 to 13% with calcium fortification. There was no change in fecal bile acid excretion. The high Ca diet significantly reduced total cholesterol 6% (5.99 to 5.66 mmol/L), LDL cholesterol 11% (4.13 to 3.67 mmol/L), and apolipoprotein B concentrations 7% when compared with the low Ca diet (P < 0.05). There was no change in HDL cholesterol or apolipoprotein A1 concentrations. Urinary calcium excretion increased from 146 to 230 mg/d when the high Ca diet was consumed. Calcium fortification was effective in lowering total and LDL cholesterol concentrations and may be an effective adjunct to cholesterol-lowering diet therapy.

Calcium chloride is a better calcium source rather than calcium carbonate for weanling pigs.

Two experiments were conducted to evaluate the effects of calcium (Ca) levels in weanling pigs (Landrace × Yorkshire × Duroc). In experiment 1, one hundred and eighty weanling pigs were randomly allotted to one of the three treatments. The treatments were low (Ca 0.60% in phase 1 and 0.50% in phase 2), standard (Ca 0.72% in phase 1 and 0.66% in phase 2), and high (Ca 0.84% in phase 1 and 0.72% in phase 2). In experiment 2, hundred and forty weanling pigs were randomly assigned to one of four treatments differing in Ca levels (high and low) and sources (CaCl2 and CaCO3) in a 2 × 2 factorial arrangement. There were 10 pigs per replicate in both experiments, with 6 replicates in each treatment, and they were conducted in two phases (phase 1, days 0–14; phase 2, days 15–28). In experiment 1, body weight (BW), average daily gain (ADG), and growth to feed ratio (G/F) increased as the Ca level decreased (p < 0.05). P digestibility was higher in the low-Ca diet group than in the high-Ca diet group (p <0.05). In experiment 2, the final BW, ADG, and G/F increased in the CaCl2 diet group compared with the case in the CaCO3 diet group (p < 0.05). The digestibility of crude protein (CP), Ca, and P was higher in the CaCl2 diet group than in the CaCO3 diet group (p < 0.05). Cl− levels were higher in the CaCl2 diet group than in the CaCO3 diet group (p < 0.05). The bicarbonate (HCO3−), base excess (BE), and electrolyte balance (EB) levels were lower in the CaCl2 diet group than in the CaCO3 diet group (p < 0.05). Hematocrit increased as the Ca level decreased (p < 0.05). The HCO3− interacted with the Ca sources and thus, affected the Ca levels (p < 0.05). Bone ash, Ca, and P were downregulated in the low-Ca diet group compared with the case in the high-Ca diet group. Overall, the low dietary Ca supplementation led to greater growth performance. Furthermore, CaCl2 appeared to be a better Ca source than CaCO3 because of the greater digestibility of CP, Ca, and P, and improved EB.

High dietary Ca and microbial phytase reduce expression of Ca transporters while enhancing claudins involved in paracellular Ca absorption in the porcine jejunum and colon.

Expression levels of genes (RT-qPCR) related to Ca and P homeostasis (transporters and claudins (CLDN)) were determined in porcine jejunal and colonic mucosa. Forty growing pigs (BW 30.4±1.3 kg) received a low and high Ca content (2.0 and 9.6 g/kg, respectively) diet with or without microbial phytase (500 FTU/kg) for 21 days. Dietary Ca intake enhanced serum Ca and alkaline phosphatase concentration and reduced P, 1,25(OH)2D3, and parathyroid hormone concentration. Jejunal TRPV5 mRNA expression was decreased (32%) with phytase inclusion only, while colonic transient receptor potential vanilloid 5 (TRPV5) mRNA was reduced by dietary Ca (34%) and phytase (44%). Both jejunal and colonic TRPV6 mRNA expression was reduced (30%) with microbial phytase. Calbindin-D9k mRNA expression was lower in colonic but not jejunal mucosa with high dietary Ca (59%) and microbial phytase (37%). None of the mRNAs encoding the Na-P cotransporters (NaPi-IIc, PiT-1, PiT-2) were affected. Jejunal, but not colonic expression of the phosphate transporter XPR1, was slightly downregulated with dietary Ca. Dietary Ca downregulated colonic CLDN-4 (20%) and -10 (40%) expression while CLDN-7 was reduced by phytase inclusion in pigs fed low dietary Ca. Expression of colonic CLDN-12 tended to be increased by phytase. In jejunal mucosa, dietary Ca increased CLDN-2 expression (48%) and decreased CLDN-10 (49%) expression, while phytase slightly upregulated CLDN-12 expression. In conclusion, compared to a Ca deficient phytase-free diet, high dietary Ca and phytase intake in pigs downregulate jejunal and colonic genes related to transcellular Ca absorption and upregulate Ca pore-forming claudins.

Interactive effect of dietary levels of calcium and 25-hydroxy vitamin D3 on the performance, serum biochemical concentration and digestibility of laying hens from 61 to 70 weeks of age.

ObjectiveThe present research was conducted to evaluate the interactive effect of dietary concentration of calcium (Ca) and 25-hydroxy vitamin D3 (25OHD3) on the performance, blood composition and digestibility of laying hens.MethodsA total of 540 Hy-line brown laying hens aged 61 to 70 weeks were randomly allotted in a 3×3 factorial arrangement, consisting of three levels of 25OHD3 (0, 25, and 50 μg/kg) and three levels of Ca (3.5%, 4.0%, and 4.5%). All diets had basal concentration of 3,000 IU/kg of vitamin D3 including the 2,800 kcal/kg of metabolic energy and 16% of crude protein.ResultsThe results showed that interactive effect (p<0.05) between Ca and 25OHD3 was such that dietary 25OHD3 linearly increased interleukin-6 at all levels of Ca inclusion. Interaction (p<0.05) occurred with the highest parathyroid hormone in laying hens that received dietary concentration of Ca (3.5%) with 25OHD3 (50 μg/kg), and Ca (4.0%) with 25OHD3 (50 μg/kg). Egg production and egg weight significantly (p<0.05) increased in the 4.5% Ca group compared to the 3.5% to 4.0% Ca groups. Egg shell thickness and tibia bone length also increased (p<0.05) in groups fed a high-Ca diet (4.0% to 4.5%). Phosphorus digestibility significantly (p<0.05) increased along with dietary Ca level. Among the tested 25OHD3 groups, higher (p<0.05) egg production and tibia thickness were present in hens fed 50 μg/kg of 25OHD3. Furthermore, Ca digestibility serum Ca and 25OHD3 were significantly increased in group offered 50 μg/kg of 25OHD3.ConclusionThe results gathered in this study indicate that dietary concentrations of 4.0% to 4.5% Ca and 50 μg/kg 25OHD3 improve the performance of hens from 61 to 70 weeks of age.

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

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

Calcium to magnesium intake ratio and non-alcoholic fatty liver disease development: a case-control study

BackgroundNon-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide. Adoption of sedentary life style and westernized diet are shown to be associated with development of NAFLD. Since previous studies suggested that calcium (Ca) to magnesium (Mg) ratio intake is associated with some chronic diseases including dyslipidemia and insulin resistance, we designed this study to find any possible association between this ratio and NAFLD development.MethodsThe NAFLD was diagnosed using Fibroscan according to a CAP cut-off value of 263 dB/m. Dietary intakes of one hundred and ninety-six patients with incident NAFLD diagnosis, and eight hundred and three controls without NAFLD were assessed using a valid food frequency questionnaire (FFQ). Dietary nutrients were calculated using Nutritionist IV software.ResultsAge of the study population (57 % female) was 43.2 ± 14.1 years. In addition, energy-adjusted daily calcium to magnesium intake ratio was 2.34 ± 0.57 and 2.73 ± 0.69 for control and case groups, respectively. In the multivariable-adjusted model, after adjustment for potential confounding variables; including, age, gender, BMI, alcohol consumption, smoking, diabetes, physical activity, energy, dietary fiber, carbohydrate, fat, and protein intakes, participants in the third (Q3) and fourth (Q4) quartile of Ca/Mg ratio intake had a greater development of incidental NAFLD compared to the lowest quartile (Q1) [(OR = 2.86; 95 % CI: 1.20–6.81), (P-value = 0.017) and (OR = 5.97; 95 % CI: 2.54–14.01), (P-value < 0.001) for Q3 and Q4 compared to the Q1, respectively]. Moreover, energy-adjusted Ca to Mg intake ratio was positively correlated with plasma level of ALT (r = 0.18; P = 0.01); contrarily, it had no correlation with plasma levels of AST.ConclusionsThe current study revealed that higher dietary Ca to Mg intake ratio is associated with a greater development of NAFLD. Further interventional studies are needed to confirm the causal relationship of the Ca/Mg ratio intake and development of NAFLD.

Perspective: Characterization of Dietary Supplements Containing Calcium and Magnesium and Their Respective Ratio—Is a Rising Ratio a Cause for Concern?

Low magnesium intakes coupled with high calcium intakes and high calcium-to-magnesium (Ca:Mg) intake ratios have been associated with increased risk for multiple chronic conditions such as cardiovascular disease and metabolic syndrome, as well as some cancers (colorectal, prostate, esophageal), and total mortality. A high dietary Ca:Mg ratio (>2.60) may affect body magnesium status while, on the other hand, high intakes of magnesium could adversely impact individuals with an exceedingly low dietary Ca:Mg ratio (<1.70). Thus, a Ca:Mg ratio range of 1.70-2.60 (weight to weight) has been proposed as an optimum range. Data from NHANES surveys have shown the mean Ca:Mg intake ratio from foods alone for US adults has been >3.00 since 2000. One-third of Americans consume a magnesium supplement with a mean dose of 146mg/d, and 35% of Americans consume a calcium supplement with a mean dose of 479mg/d. Our review of Ca:Mg ratios in dietary supplements sold in the United States and listed in NIH's Dietary Supplement Label Database (DSLD) found a mean ratio of 2.90 across all calcium- and magnesium-containing products, with differences by product form. The ratios ranged from a low of 0.10 in liquid products to a high of 48.5 in powder products. Thirty-one percent of products fell below, 40.5% fell within, and 28.3% fell above the ratio range of 1.70-2.60. Our findings of calculated Ca:Mg ratios from dietary supplements coupled with food-intake data suggest that, in individuals with high calcium intakes from diet and/or supplements, magnesium supplementation may be warranted to establish a more favorable dietary Ca:Mg ratio in their total diet. Additional research may provide greater insight into whether the Ca:Mg ratio is a biomarker of interest for moderating chronic disease and which population groups may derive benefit from moderating that ratio.

Biochemical and molecular investigation of oxidative stress associated with urolithiasis induced by increased dietary calcium or protein in chickens.

This study was carried out to evaluate the effects of induced urolithiasis by high dietary calcium (Ca) or protein levels on biochemical analyte levels, redox status, selected inflammatory cytokines and histopathology in chickens. A total of 90 one-day-old white Hy-Line chicks were fed basal control diets containing 20% crude protein (CP) and 1% Ca until they reached 44days of age. After that, the birds were divided into three groups (30 birds per group). All management factors (light, temperature, ventilation, stock density and diet) were identical among the three groups throughout the study except for the dietary Ca and protein percentages. Group I was fed a control diet containing 20% CP and 1% Ca, group II was fed a high-Ca diet containing 5% Ca, and group III was fed a high-protein diet containing 25% CP. Our findings clearly demonstrated that dietary imbalance (caused by high-Ca or high-CP levels) per se in chickens was physiologically harmful, as it was accompanied by post-mortem lesions; biochemical, redox status and histopathological alterations; and upregulation of inflammatory cytokines (interleukin (IL)-1β and IL-6). In particular, the birds fed the high-Ca diet clearly exhibited the most obvious alterations in most of the endpoints. In conclusion, this study constitutes the first extensive investigation of the effects of high-Ca or high-protein diets induced urolithiasis on growth performance, redox status, inflammatory cytokine levels and pathological characterization in chickens.

Interactive effect of dietary calcium and phytase on broilers challenged with subclinical necrotic enteritis: part 2. Gut permeability, phytate ester concentrations, jejunal gene expression, and intestinal morphology

Calcium has the capacity to interact with phytate-P to form Ca-phytate complexes and decrease the ability of exogenous phytase to degrade phytic acid. This study investigated the hypothesis that high dietary Ca would impair gut permeability, phytate esters (inositol x-phosphate, IPx: IP3, IP4, IP5, and IP6) degradation, jejunal gene expression, and intestinal morphology. Ross 308 day-old male broilers (n = 768) were distributed into 48-floor pens each housing 16 birds in a factorial arrangement. Factors were NE challenge—no or yes; phytase level of 500 or 1,500 FTU/kg, and Ca level 0.6 or 1.0% starter, 0.5 or 0.9% grower, 0.4 or 0.8% finisher with available P in each phase. Challenged birds were gavaged with 3 field strains of Eimeria on day 9 and 108 CFU per mL of Clostridium perfringens Strain EHE-NE18 on day 14 and day 15. A phytase × Ca interaction was observed in the ileum for IP3 (P < 0.01), IP4 (P < 0.05), and IP6 (P < 0.01). The IP3 and IP4 concentrations were similar for both doses of phytase in the presence of low Ca, but with high Ca, both increased significantly but to a greater extent when the high dose of phytase was used. While IP6 concentrations were low and similar between both doses of phytase at low Ca levels, increasing dietary Ca levels increased IP6 concentrations regardless of phytase dose, but the effect was greater in the low phytase diet. A phytase × Ca interaction was detected for vitamin D receptor (VDR) (P < 0.05) expression where bird fed low phytase and low Ca recorded the highest expression of VDR, all other treatments being equivalent. The challenge decreased crypt depth to villus height ratio (P < 0.001). Challenge birds had higher fluorescein isothiocyanate dextran (P < 0.05) in blood compared with unchallenged birds. Thus, high Ca and high phytase, while not the best for IP6 destruction, did not lead to huge reductions in indicators of gut health.

Interactive effect of 2 dietary calcium and phytase levels on broilers challenged with subclinical necrotic enteritis: part 1—broiler performance, gut lesions and pH, bacterial counts, and apparent ileal digestibility

This study investigated the hypothesis that high dietary calcium (Ca) would stimulate necrotic enteritis (NE) and reduce performance, gut health, and nutrient digestibility, and if increased, phytase would reduce NE. Ross 308 male broilers (n = 768) were randomly distributed to 8 treatments in a factorial arrangement. Factors were NE challenge (no or yes), phytase level (500 or 1,500 FTU/kg using 500 FTU/kg matrix values), and Ca level (0.6 or 1.0% starter, 0.5 or 0.9% grower, 0.4 or 0.8% finisher) with the same level of available P (0.40 S, 0.35 G, and 0.35 F). There were 48 pens, 16 birds per pen and 6 replications. Half of the birds were challenged with Eimeria spp on day 9 and 108 CFU per mL of Clostridium perfringens strain EHE-NE18 on day 14 and 15. Gain was higher in birds fed high phytase on day 14 (P < 0.01), day 21 (P < 0.01), day 28 (P < 0.01), and day 35 (P < 0.01). Birds fed high phytase had greater livability on day 21 (P < 0.01). Ca was more digestible in high-Ca diets on day 16, and an NE × Ca interaction (P < 0.05) showed this effect to be more pronounced in unchallenged than in challenged birds. A challenge × Ca interaction for apparent ileal digestibility (AID) of crude protein (CP) (P < 0.05) indicated lower AID of CP in challenged birds fed high Ca. The challenge decreased AID of Ca (P < 0.01). Ca level had no impact on C. perfringens count, but it decreased Lactobacillus (P < 0.05) and Bifidobacteria (P < 0.05) populations in the ceca. High dietary Ca decreased feed conversion ratio. Overall (42 D), the highest WG was observed in unchallenged birds fed high Ca and high phytase with the lowest WG observed in NE-challenged birds fed low Ca and low phytase. The results suggest that full matrix values for high doses of phytase may be appropriate during NE challenge.

Negative dietary cation-anion difference and amount of calcium in prepartum diets: Effects on milk production, blood calcium, and health

Acidogenic prepartum diets with negative dietary cation-anion difference (DCAD) induce compensated metabolic acidosis, which stimulates calcium (Ca) mobilization before calving and decreases clinical and subclinical hypocalcemia postpartum. This strategy is often combined with limiting dietary Ca concentrations, which historically has been used to mobilize Ca prepartum to prepare cows for lactation. Supplemental dietary Ca in combination with a negative DCAD formulation that does not reverse the effect of compensated metabolic acidosis may be beneficial. Our objective was to determine the effects of prepartum dietary strategies on dry matter intake (DMI), milk production, peripartal Ca status, and health during the transition period in multiparous Holstein cows (n = 81). Treatments during the last 28 d before calving were: (1) positive DCAD diet, +6 mEq/100 g of DM, target urine pH >7.5, low dietary Ca (0.40% DM; CON); (2) negative DCAD diet, -24 mEq/100 g of DM, target urine pH 5.5 to 6.0, low dietary Ca (0.40% DM; ND); or (3) negative DCAD diet, -24 mEq/100 g of DM, target urine pH 5.5 to 6.0, , high dietary Ca (2.0% DM; NDCA). Preplanned treatment contrasts were: (1) CON versus (ND and NDCA), and (2) ND versus NDCA. Individual DMI were recorded daily. Cows were milked 3 times daily, with individual DMI and milk yield summarized by week. Whole blood sampled at calving and 24 h, 48 h, and 4 d after calving was analyzed for ionized Ca concentration, and serum was analyzed for total Ca. Prepartum urine pH for cows fed ND or NDCA averaged 5.7, whereas cows fed CON remained >7.5. During the 3 wk before calving, cows fed CON had greater DMI than cows fed ND or NDCA, with NDCA greater than ND. Postpartum DMI (% of body weight) tended to be less for cows fed CON than for those fed ND or NDCA prepartum. Thresholds for subclinical hypocalcemia were ionized Ca <1.0 mM at 24 h, and total Ca ≤2.125 mM at 48 h after calving. On average, blood Ca for cows fed CON indicated subclinical hypocalcemia, whereas blood Ca for cows fed ND or NDCA was greater than subclinical hypocalcemia thresholds for both ionized Ca and total Ca. No milk production differences were detected. Cows fed CON had an elevated adverse health score (calculated by assigning numerical values to recorded health events) and tended to have an elevated somatic cell count during the fresh period compared with cows fed ND or NDCA. Overall, an acidogenic diet prepartum without or with high Ca improved postpartum Ca status and health. Supplementation of additional Ca to the acidogenic diet had little effect.

Ca:Mg Ratio, APOE Cytosine Modifications, and Cognitive Function: Results from a Randomized Trial.

Deterioration of ionized calcium (Ca2+) handling in neurons could lead to neurodegenerative disease. Magnesium (Mg) antagonizes Ca during many physiologic activities, including energy metabolism and catalyzation of demethylation from 5-methylcytosine(5-mC) to 5-hydroxymethylcytosine(5-hmC). To test the hypothesis that actively reducing the Ca:Mg intake ratio in the diet through Mg supplementation improves cognitive function, and to test whether this effect is partially mediated by modified cytosines in Apolipoprotein E (APOE). This study is nested within the Personalized Prevention of Colorectal Cancer Trial (PPCCT), a double-blind 2×2 factorial randomized controlled trial, which enrolled 250 participants from Vanderbilt University Medical Center. Target doses for both Mg and placebo arms were personalized. Among those aged > 65 years old who consumed a high Ca:Mg ratio diet, we found that reducing the Ca:Mg ratio to around 2.3 by personalized Mg supplementation significantly improved cognitive function by 9.1% (p = 0.03). We also found that reducing the Ca:Mg ratio significantly reduced 5-mC at the cg13496662 and cg06750524 sites only among those aged > 65 years old (p values = 0.02 and 0.03, respectively). Furthermore, the beneficial effect of reducing the Ca:Mg ratio on cognitive function in those aged over 65 years was partially mediated by reductions in 5-mC levels (i.e., cg13496662 and cg06750524) in APOE (p for indirect effect = 0.05). Our findings suggest that, among those age 65 and over with a high dietary Ca:Mg ratio, optimal Mg status may improve cognitive function partially through modifications in APOE methylation. These findings, if confirmed, have significant implications for the prevention of cognitive aging and Alzheimer's disease.Clinical Trial Registry number and website: #100106 https://clinicaltrials.gov/ct2/show/NCT03265483.

Interactive effect of dietary calcium and phytase on broilers challenged with subclinical necrotic enteritis: 3. Serum calcium and phosphorus, and bone mineralization

Calcium is chelated by phytic acid and forms phytate-mineral complexes reducing Ca availability and the ability of phytase to hydrolyze phytate. An increased Ca concentration in the gut favors the activity of Clostridium perfringens (C. perfringens). Therefore, it was hypothesized that high dietary calcium with high dietary phytase would decrease serum Ca and P and bone mineralization during necrotic enteritis occurrence. A total of 768 one-day-old Ross 308 male chicks were randomly allocated to 8 treatments with 6 replicate pens, each housing 16 birds. A 2 × 2 × 2 factorial arrangement of treatments was applied: dietary Ca (0.6 or 1.0%), phytase (500 or 1,500 FTU/kg), and challenge (no or yes). Half of the birds (384) were challenged with Eimeria spp. on day 9 and C. perfringens strain EHE-NE18 on day 14 and 15. Blood was collected from 2 birds per pen to determine Ca, P, and parathyroid hormone in the serum. The middle toe, tibia, and femur were excised from 2 birds per pen on day 16 and 29 for determination of ash, breaking strength (BS), and mineral concentration. The challenge decreased (P < 0.05) serum Ca+ in birds regardless of dietary Ca level (day 16). There was a challenge × Ca interaction (P < 0.05) for tibial BS (day 16), with challenge being more severe in birds fed high Ca than low Ca diets. A challenge × phytase interaction (P < 0.05) was present for femur ash (day 16), with high phytase only increasing ash in challenged birds. The challenge decreased (P < 0.05) the BS of femur and tibia at each time point. Birds fed high dietary Ca had lower tibial Mg (P < 0.001), Fe (P < 0.001), Na (P < 0.001), and Zn (P < 0.05) concentrations (day 29). Altogether, high dietary Ca and phytase improved bone mineralization showing that attention to Ca and P nutrition and phytase matrix values is warranted when high levels of phytase are used.

Effect of Silicon Supplementation in Diets with Different Calcium Levels on Balance of Calcium, Silicon and Magnesium, and Bone Status in Growing Female Rats.

Silicon (Si) is important for the growth and development of bone and connective tissues. This study aimed to evaluate the effect of Si supplementation on the balance of calcium (Ca), Si, magnesium (Mg), and bone status in growing female rats fed diets with different Ca levels. Sixty 6-week-old female Wistar rats were divided into 6 groups and fed diets with different levels of Ca (0.1, 0.5, 1.5%) and Si (5, 500ppm) for 10weeks. Experimental groups were as follows: Low-Ca group (LCa; 0.1% Ca), Low-Ca with Si supplementation group (LCaSi), adequate-Ca group (ACa; 0.5% Ca), adequate-Ca with Si supplementation group (ACaSi), high-Ca group (HCa; 1.5% Ca), and high-Ca with Si supplementation group (HCaSi). Si retention significantly increased by Si supplementation and significantly decreased by high-Ca diet (1.5%). Mg retention significantly decreased by high-Ca diet. Serum Ca and Si contents were not significantly different among the 6 groups. Low-Ca diet significantly increased serum osteoclain and C-telopeptide cross-link of type 1 collagen (CTx) levels, and Si supplementation significantly decreased CTx levels in the low-Ca diet group. Bone mineral density was significantly increased along with Ca increase in the diet, but was decreased by Si supplementation in the high-Ca diet. Rats in the adequate-Ca diet group had a significant increase in strength of tibia after Si supplementation. These findings indicate that the effect of Si supplementation on bone metabolism may differ according to the Ca-intake level in growing females. Although further research is needed, when supplementing Si to improve bone health in growing females, Ca-intake level should be considered.

Volatile basic nitrogen measurement in digesta using a Berthelot reaction in automated Skalar instrumentation.

The undigested nitrogenous fraction entering the hindgut of chickens is further metabolized by microbiota present producing volatile basic metabolites including amines and ammonia (NH3). Ammonia increases pH and may result in overgrowth of Clostridium perfringens further producing toxic metabolites that cause dysbacteriosis or necrotic enteritis (NE). There are few reports in chickens examining the production and concentration of nitrogenous metabolites in the hindgut. A Berthelot reaction using an automated flow, reaction and spectrophotometric instrumentation to detect volatile basic nitrogen in the form of NH3 from cecal contents was carried out. In the Berthelot reaction, NH3 in the sample is chlorinated to monochloramine using dichloroisocyanuric acid that then reacts with salicylate to form 5-aminosalicylate which is stable under alkaline conditions (pH 12 to 13). After oxidation and oxidative coupling, a coloured complex is formed that can be measured at 660 nm. Cecal contents were collected and pooled from 2 birds per pen with 48 pens total in each of 3 experiments and stored in sealed containers at −20 °C prior to analysis. Experiment 1 compared samples collected from birds fed either no meat and bone meal (MBM) or 6% MBM from d 0 to 14, and samples collected from birds fed either no MBM or 5% MBM from d 14 to 42. All birds were challenged with Eimeria on d 9 and C. perfringens on d 14 and 15. Experiment 2 compared cecal contents from birds fed either 0.5% or 0.9% calcium (Ca), and Exp. 3 compared unchallenged with NE challenge on d 16 and 29. Results demonstrated an increase (P < 0.05) in cecal NH3 as a result of feeding MBM or high dietary Ca with a corresponding increase in pH. Birds experimentally challenged with NE had lower cecal NH3 and lower pH on d 16 but not d 29 compared to unchallenged controls. It was concluded that the Berthelot method using Skalar instrumentation equipment is suitable to measure the concentration of volatile nitrogen as NH3 in cecal contents of chickens.