The effects of second messenger cAMP and its relative components on the contraction of uterine smooth muscle of rat.

OBJECTIVE: To investigate the effect of second messenger pathways on the uterine smooth muscle contraction and their associated mechanisms, and compare the evaluation methods. MATERIALS AND METHODS: Preparation of uterine smooth muscle strips from healthy pregnant 18-21 d SD and non-pregnant rats. When the contraction of muscle strips was stable, we conducted gradient administration: PDE4 inhibitors (Z90), prostaglandin PGE2, adenylate cyclase inhibitor (SQ 22,530), cAMP analogs (dbcAMP) and AMPK agonists (AICAR), solvent dimethyl sulfoxide (DMSO) as controlled. Gradient administration of acetylcholine (Ach) and oxytocin (oxytocin) induced the contraction of muscle strips. The tension transducer and biological information collecting system were applied to record the changes, including duration, dilation tension, contraction tension, peak height, and mean tension, before and after different administration. Principal components analysis was adopted to evaluate the five changes. RESULTS: SQ 22,530, DMSO, cAMP alone had no significant effect on the contraction of uterine smooth muscle; Z90 can inhibit the spontaneous contraction of pregnant uterine smooth muscle strips; dbcAMP and AICAR can antagonize acetylcholine and oxytocin-induced the contraction of pregnant uterine smooth muscle strips. Z90, SQ 22,530 + Z90, dbcAMP, AICAR can inhibit the uterine contraction peak, diastolic amplitude, average muscle tone and contraction duration of the pregnant uterine smooth muscle in a concentration-dependent manners. At the same time, we compared the parameters, which reflect the contraction of uterine smooth muscle, and conduct main components analysis to determine the effect of the drugs. CONCLUSIONS: The second messenger cAMP and its related components ATP, 5'- AMP, AC, PDE, PKA, and AMPK can affect the uterine smooth muscle contraction via related signaling pathway in rats, and principal components analysis can be adopted to evaluate the smooth muscle relaxant.

Continual feeding of two types of microalgal biomass affected protein digestion and metabolism in laying hens.

A 14-wk study was conducted to determine the nutritional efficacy and ssmetabolic impact of 2 types of microalgal biomass as alternative protein sources in laying hen diets. Shaver hens (total = 150 and 26 wk old) were fed 1 of 5 diets: a control or a defatted green microalgal biomass (DG; Desmodesmus spp.) at 25% and a full-fatted diatom biomass (FD; Staurosira spp.) at 11.7% inclusion with or without protease. This experiment consisted of 5 replicates per treatment and each replicate contained 6 hens individually reared in cages (1 hen for biochemical data/replicate). Despite decreased ADFI (P = 0.03), hens fed DG or FD had final BW, overall hen-day egg production, and egg quality similar to the controls. Feeding DG or FD did not alter plasma concentrations of insulin, glutamine, and uric acid or alkaline phosphatase activity at wk 8 or 14 but decreased plasma 3-methyhistine concentrations (P = 0.03) and tartrate-resistant acid phosphatase (TRAP) activities (P < 0.001) at wk 14 and improved (P = 0.002) ileal total AA digestibility. Although DG or FD exhibited moderate effects on intestinal brush border protease activities and mRNA levels of duodenal transporters Pept1, Lat1, and Cat1, both substantially enhanced (P < 0.05) phosphorylation of hepatic protein synthesis key regulator S6 ribosomal protein (S6) and the ratio of phospho-S6 to S6 in the liver of hens. However, DG and FD manifested with different impacts on weights of egg and egg albumen, proteolytic activity of jejunal digesta, plasma TRAP activity, ileal total AA digestibility, and several intestinal genes and hepatic proteins. Supplemental protease in the DG and FD diets produced mixed effects on a number of measures. In conclusion, our findings revealed the feasibility of including greater levels of microalgal biomass as a source of feed protein for laying hens and a novel potential of the biomass in improving dietary protein digestion and body protein metabolism than previously perceived.

Lipopolysaccharide and interferon-gamma-induced nitric oxide production and protein oxidation in mouse peritoneal macrophages are affected by glutathione peroxidase-1 gene knockout.

This study investigated the role of glutathione peroxidase-1 (GPX1) in protein oxidation in peritoneal macrophages. Macrophages isolated from both wild-type (WT) and GPX1 knockout (KO) mice were activated by lipopolysaccharide (LPS, 1 microg/ml) and interferon-gamma (IFN, 10 U/ml for 24 or 48 h in the presence or absence of 1 microM diquat (DQ), 250 microM aminoguanidine (AG, an inhibitor of inducible nitric oxide synthase), and (or) 100 microM diethyldithiocarbamate (DETC, an inhibitor of Cu,Zn-SOD). In the KO macrophages, there was no protein band detected by Western blot with anti-GPX1 antibody and 98% reduction in total GPX activity compared with WT cells. Nitric oxide (NO) synthesis was greatly enhanced after 24 h by GPX1 knockout and DQ, but inhibited by AG or DETC. Protein carbonyl formation in total cell extract was clearly associated with NO synthesis as higher levels of protein carbonyl were detected in activated KO than WT macrophages, and DQ enhanced slightly while AG or DETC virtually blocked its formation. A similarly marginal effect of GPX1 KO was observed on protein nitration. The LPS/IFN/DQ-induced DNA fragmentation was blocked by AG, but not by DETC. Cell viability at 48 h was decreased by the LPS/IFN activation and further reduced by the addition of DQ, but restored by AG. In conclusion, GPX1 affects the NO production in activated peritoneal macrophages and protects these cells against NO-associated protein oxidation.

Knockout of cellular glutathione peroxidase gene renders mice susceptible to diquat-induced oxidative stress.

Two experiments were conducted to determine the protection and the underlying mechanisms of cellular glutathione peroxidase (GPX1) against lethal, acute oxidative stress induced by an intraperitoneal injection of 24 mg diquat/kg body weight. In experiment 1, mortality and survival times were compared among selenium (Se)-adequate or deficient GPX1 knockout mice [GPX1(-/-)] and wild-type mice (WT). In experiment 2, mice from these four groups were euthanized at 0, 1, 2, and 3 h after the injection of diquat to elucidate the time course of oxidative events. The stress produced 100% mortality in all of the groups except for the Se-adequate WT, which were euthanized on day 7 for analysis. The Se-deficient WT and the Se-adequate GPX1(-/-) had similar survival times (4.1 and 3.9 h), which were longer (p < .05) than that of the Se-deficient GPX1(-/-) (2.4 h). However, these three GPX1-deficient groups had higher levels (p < .05) of hepatic F2-isoprostanes and carbonyl contents and/or plasma alanine aminotransferase activities than those of the Se-adequate WT. The diquat-induced formations of hepatic F2-isoprostanes in these animals peaked at 1 h and preceded the rise of plasma alanine aminotransferase in the Se-adequate GPX1(-/-). Responses of hepatic superoxide dismutase activities to the diquat treatment were affected by the GPX1 level. In conclusion, GPX1 is the major selenoprotein to protect mice against the lethal oxidative stress induced by diquat.

Comparison of age-related differences in expression of phospholipid hydroperoxide glutathione peroxidase mRNA and activity in various tissues of pigs.

Phospholipid hydroperoxide glutathione peroxidase (PHGPX) is the second identified Se-dependent intracellular glutathione peroxidase (PHGPX) that reduces phospholipid hydroperoxides. The objective of this study was to determine the developmental regulation of PHGPX expression in tissues of neonatal, weanling and finishing pigs (Sus scrofa) compared with the expression of the classic Se-dependent cellular glutathione peroxidase (GPX) and the Se-independent enzyme, glutathione S-transferase (GST). Eight different tissues were collected from Se-adequate male pigs aged 1, 28 and 180 days, and supernatant of the tissue homogenate was assayed for PHGPX, GPX and GST activities by using phosphatidylcholine hydroperoxide, hydrogen peroxide and 1-chloro-2,4-dinitrobenzene as substrate, respectively. Total RNA was isolated from four tissues and assayed for PHGPX mRNA expression. Both mRNA and activity expression of PHGPX in most assayed tissues was increased as pigs became older (P < 0.05), but increases in PHGPX mRNA levels between ages did not fully account for all changes in activity. Expression of GPX activity was increased more than that of PHGPX between day 1 and day 28 (P < 0.0001). Expression of GST activity in various tissues was also affected by age (P < 0.01) but lacked a consistent relationship with the changes in GPX and PHGPX activity. Tissue-specific patterns of developmental expression of these enzymes may be related to the susceptibility of organs to pro-oxidant injuries. In conclusion, expression of PHGPX mRNA and activity in various tissues of pigs is developmentally increased over ages, and the pattern is somewhat different from that of GPX.

Glutathione peroxidase-1 gene knockout on body antioxidant defense in mice.

To determine the in vivo role of cellular glutathione peroxidase (E.C.1.11.1.9, GPX1), we challenged the GPX1 knockout [GPX1(-/-)], the GPX1 overexpressing [GPX1(+)], and their respective wild-type (WT) mice of different Se and vitamin E status with acute oxidative stress. After these mice were injected with pro-oxidants paraquat or diquat at 12 to 125 mg/kg of body weight, their survival rate and time were a function of their GPX1 activity levels. The GPX1 protection was associated with attenuation of NADPH and NADH oxidation, protein carbonyl and F(2)-isoprostanes formation, and alanine transaminase release in various tissues, and was irreplaceable by high levels of dietary vitamin E or other selenoproteins. The GPX1 expression was also protective against moderate oxidative stress induced by low levels of paraquat or diquat, particularly in the Se-deficient mice. Alteration of GPX1 expression showed no impact on the expression of other selenoproteins and antioxidant enzymes in unstressed mice. Total Se content in liver of the Se-adequate GPX1(-/-) mice was reduced by 60% the WT controls. In conclusion, normal expression of GPX1 is essential and overexpression of GPX1 is beneficial to protect mice against acute oxidative stress.

Knockout of cellular glutathione peroxidase affects selenium-dependent parameters similarly in mice fed adequate and excessive dietary selenium.

This study was to determine whether or not effects of the cellular glutathione peroxidase (GPX1) knockout on several Se-dependent parameters in mice were tissue, dietary Se concentration, and selenoprotein specific. A 2 x 3 factorial experiment was conducted with 18 GPX1 knockout mice [GPX1(-)] and 18 controls (3 weeks old, half males and females). These mice were fed a torula yeast diet supplemented with all-rac-alpha-tocopheryl acetate (50 mg/kg of feed) and Se (sodium selenite) at 0, 0.5, or 3.0 mg/kg of feed for 6 weeks. Both kidney GPX1 mRNA levels and liver, kidney, lung, and testis total GPX activities, assayed using hydrogen peroxide, were affected (p < 0.001) by the GPX1 knockout and dietary Se concentrations, whereas kidney extracellular or plasma GPX (GPX3) mRNA levels and phospholipid hydroperoxide GPX (GPX4) activities in the four tissues were affected (p < 0.001) by only dietary Se concentrations. Total GPX activity in testis was reduced approximately 90% (p < 0.01) by the GPX1 knockout. Neither the GPX1 knockout nor the dietary Se concentrations affected mRNA levels of GPX4 in testis or selenoprotein P in kidney. Total liver Se concentrations were not different between the GPX1(-) and control mice at 0 mg Se/kg of feed, but were reduced (p < 0.01) by 61 and 64% in the GPX1(-) mice at 0.5 and 3.0 mg Se/kg of feed, respectively. These results not only confirm the independent expression of GPX3, GPX4, and selenoprotein P from that of GPX1, but also show similar effects of the GPX1 knockout on Se-dependent parameters in mice between different dietary Se concentrations, tissues, and selenoproteins.

Supplementing corn-soybean meal diets with microbial phytase linearly improves phytate phosphorus utilization by weanling pigs.

Two experiments were conducted with weanling pigs to determine the effectiveness of a dietary supplement of Aspergillus niger phytase in improving the availability of phytate-P in corn-soybean meal diets without supplemental inorganic P. Experiment 1 consisted of two P and Ca balance trials and two feeding trials. Twelve pigs (8.18 +/- .44 kg BW) were housed individually in stainless steel metabolism cages. Six pigs received 750 phytase units (PU)/g of basal diet and the other six pigs received the basal diet without supplemental phytase as control. In Exp. 2, 96 pigs (8.81 +/- .75 kg BW) were allotted to 16 partially slotted floor pens and their basal diets were supplemented with either 0, 250, 500, or 750 PU/g for 4 wk. Individual pig weights and pen feed consumption were measured weekly. Blood samples were taken from all pigs at the end of each trial in Exp. 1 and from three pigs per pen weekly in Exp. 2 to measure serum (plasma) inorganic P (P) and Ca concentrations and alkaline phosphatase (AP) activities. The results of Exp. 1 indicated that dietary phytase increased P retention by 50% (P < .0001) and decreased fecal P excretion by 42% (P < .0001). Pigs that received dietary phytase had serum P and Ca concentrations and serum AP activities that were nearly normal, whereas control pigs had values indicative of a moderate P deficiency. Favorable effects of phytase disappeared when the phytase was removed from the diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Supplementing corn-soybean meal diets with microbial phytase maximizes phytate phosphorus utilization by weanling pigs.

Two experiments were conducted with crossbred weanling pigs to determine the optimal dietary supplement of Aspergillus niger phytase activity to a low-P, corn-soybean meal basal diet (BD). In Exp. 1, 50 pigs (7.61 +/- .56 kg BW) received the BD supplemented with 750, 1,050, 1,250, or 1,350 phytase units (PU)/g, or .21% P as mono-dibasic calcium phosphate (MDCaP) for 4 wk. In Exp. 2, 12 pigs (6.39 +/- .74 kg BW) were individually housed in metabolism cages and received BD, BD plus the optimal phytase activity (1,200 PU/g), or BD plus .21% P as MDCaP for 2 wk. In Exp. 1, additions of phytase > 1,050 PU/g of BD did not improve ADG, ADFI, gain/feed, or plasma AP activity. Quadratic relationships between dietary phytase activity and these measures were found and their stationary points were at approximately 1,200 PU/g of BD. Estimated maximum responses of these measures in pigs fed phytase were > or = 90% compared with MDCaP. Pigs fed 1,250 PU/g of BD maintained normal plasma P and Ca concentrations. In Exp. 2, pigs that received 1,200 PU/g of BD utilized dietary P more effectively (P < .05) than pigs fed the BD or the BD plus MDCaP. Although they consumed 44% less P per day, these pigs retained only 7% less P than pigs that received MDCaP. One thousand units of phytase activity supported retention of 1.1 mg of P from the BD, and this level of phytase supplementation was equivalent in effect to .91 mg of P from MDCaP.(ABSTRACT TRUNCATED AT 250 WORDS)

A new phytase expressed in yeast effectively improves the bioavailability of phytate phosphorus to weanling pigs.

We have recently expressed a new phytase enzyme in a yeast system. Three experiments with a total of 140 weanling crossbred pigs were conducted to examine the efficacy of this enzyme in improving the bioavailability of phytate-P in corn-soybean meal diets to young pigs. Experiment 1 compared the efficacy of this new phytase with a commercially available phytase (Natuphos, BASF) for 4 wk at an inclusion level of 1,200 U/kg of diet. Experiment 2 compared the responses of pigs to four doses of the new phytase supplementation (300, 600, 900, and 1,200 U/kg of diet) for 4 wk. Experiment 3 compared the efficacy of this new phytase and Natuphos at a marginally optimal dose (700 U/kg of diet) for 5 wk. A group of pigs were fed the P-deficient basal diet as a negative control in Exp. 1, and a group of pigs were fed the basal diet plus .17 or .22% inorganic P as a positive control in all experiments. In Exp. 1, pigs fed the two sources of phytase had similar ADG (564 vs 567 g), gain/feed (.597 vs .589), plasma inorganic P concentrations (8.9 vs 8.4 mg/dL), and mobility scores (4.25 vs 4.46) that were higher (P < .05) than those of the negative control. In Exp. 2, plasma inorganic P concentration was a fairly linear response to the phytase dose (r > .83) at wk 1 and 2. Overall ADG of pigs also tended to increase with the phytase dose (P = .15). In Exp. 3, pigs fed the two sources of phytase had ADG (483 vs 506 g) similar to that of the positive control (508 g). These two groups also had similar plasma inorganic P concentrations (7.7 vs 7.4 mg/dL) that were lower (P < .05) than those of the positive control group (9.7 mg/dL). There was no significant effect of dietary treatments on ADFI in all three experiments. In conclusion, our new phytase was as effective as Natuphos, at the inclusion level of 700 or 1,200 U/kg of a P-deficient, corn-soybean meal diet, in improving phytate-P utilization by young pigs.

Effectiveness of an experimental consensus phytase in improving dietary phytate-phosphorus utilization by weanling pigs.

Consensus phytase is a new biosynthetic, heat-stable enzyme derived from the sequences of multiple homologous phytases. Two experiments were conducted to determine its effectiveness, relative to inorganic P and a mutant enzyme of Escherichia coli phytase (Mutant-EP), in improving dietary phytate-P availability to pigs. In Exp. 1, 36 pigs (3 wk old, 7.00 +/- 0.24 kg of BW) were fed a low-P corn-soybean meal basal diet plus consensus phytase at 0, 250, 500, 750, 1,000, or 1,250 U/kg of feed for 5 wk. Plasma inorganic P concentration, plasma alkaline phosphatase activity, bone strength, and overall ADG and gain:feed ratio of pigs were improved (P < 0.05) by consensus phytase in both linear (R2 = 0.20 to 0.70) and quadratic (R2 = 0.30 to 0.70) dose-dependent fashions. In Exp. 2, 36 pigs (4 wk old, 9.61 +/- 0.52 kg BW) were fed the basal diet + inorganic P at 0.1 or 0.2%, consensus phytase at 750 or 450 U/kg of feed, Mutant-EP at 450 U/kg of feed, or 225 U consensus + 225 U Mutant-EP/kg of feed. Pigs fed 750 U of consensus phytase or 450 U of Mutant-EP/kg feed had plasma inorganic concentrations and bone strength that fell between those of pigs fed 0.1 or 0.2% inorganic P. These two measures were 16 to 29% lower (P < 0.05) in pigs fed 450 U of consensus phytase/kg of feed than those of pigs fed 0.2% inorganic P. Plasma inorganic P concentrations were 14 to 29% higher (P < 0.05) in pigs fed Mutant-EP vs. consensus phytase at 450 U/kg at wk 2 and 3. In conclusion, the experimental consensus phytase effectively releases phytate P from the corn-soy diet for weanling pigs. The inorganic P equivalent of 750 U of consensus phytase/kg of feed may fall between 0.1 and 0.2%, but this requires further determination.

Calcium level affects the efficacy of supplemental microbial phytase in corn-soybean meal diets of weanling pigs.

A 2 x 2 x 2 factorial experiment was conducted with 64 pigs (4 wk old, 8.04 +/- .50 kg BW) to determine the effect of various dietary concentrations of Ca, vitamin D, and microbial phytase (Aspergillus niger) on phytate-P utilization. A low-P, corn-soybean meal diet was supplemented with two levels of phytase (unit/gram), 750 (suboptimal) and 1,200 (optimal); of vitamin D (international unit/kilogram), 660 (normal) and 6,660 (high); and of Ca (percentage), .4 (low) and .8 (normal). Pen feed consumption and individual pig weights, plasma inorganic P and Ca concentrations, and plasma alkaline phosphatase (AP) activity were measured at d 10, 20, and 30. The normal dietary Ca concentration had an adverse effect (P < .05) on all the response measures. The depressive effect of the normal dietary Ca on performance was greater (P < .05) at the normal vitamin D level or at the optimal phytase level than at the other levels of these two factors. The elevation in plasma AP activity in pigs fed the normal dietary Ca was greater (P < .05) at the suboptimal than at the optimal phytase level. The decreases in plasma inorganic P concentration and increases in plasma Ca concentration associated with the normal dietary Ca were substantial. In conclusion, the normal level of Ca in the diet greatly reduced the efficacy of supplemental phytase. Raising vitamin D in the diet partially offset this adverse effect but did not produce further improvement when the Ca level was low.

Phytase improves iron bioavailability for hemoglobin synthesis in young pigs.

Dietary phytase supplementation improves bioavailabilities of phytate-bound minerals such as P, Ca, and Zn to pigs, but its effect on Fe utilization is not clear. The efficacy of phytase in releasing phytate-bound Fe and P from soybean meal in vitro and in improving dietary Fe bioavailability for hemoglobin repletion in young, anemic pigs was examined. In Exp. 1, soybean meal was incubated at 37 degrees C for 4 h with either 0, 400, 800, or 1,200 units (U) of phytase/kg, and the released Fe and P concentrations were determined. In Exp. 2, 12 anemic, 21-d-old pigs were fed either a strict vegetarian, high-phytate (1.34%) basal diet alone, or the diet supplemented with 50 mg Fe/kg diet (ferrous sulfate) or phytase at 1,200 U/kg diet (Natuphos, BASF, Mt. Olive, NJ) for 4 wk. In Exp. 3, 20 anemic, 28-d-old pigs were fed either a basal diet with a moderately high phytate concentration (1.18%) and some animal protein or the diet supplemented with 70 mg Fe/kg diet, or with one of two types of phytase (Natuphos or a new phytase developed in our laboratory, 1,200 U/kg diet) for 5 wk. In Exp. 2 and 3, diets supplemented with phytase contained no inorganic P. In Exp. 1, free P concentrations in the supernatant increased in a phytase dose-dependent fashion (P<.05), whereas free Fe concentrations only increased at the dose of 1,200 U/kg (P<.10). In Exp. 2 and 3, dietary phytase increased hemoglobin concentrations and packed cell volumes over the unsupplemented group; these two measures, including growth performance, were not significantly different than those obtained with dietary supplemental Fe. In conclusion, both sources of phytase effectively degraded phytate in corn-soy diets and subsequently released phytate-bound Fe from the diets for hemoglobin repletion in young, anemic pigs.

Efficacy of an E. coli phytase expressed in yeast for releasing phytate-bound phosphorus in young chicks and pigs.

Four chick trials and one pig trial were conducted to investigate the phosphorus-releasing efficacy oftwo commercial phytase enzymes (Natuphos and Ronozyme) and an experimental E. coli phytase enzyme (ECP) when added to corn-soybean meal diets containing no supplemental inorganic P (iP). In the 13- or 14-d chick trials, three or four graded levels of iP (0, 0.05,0.10,0.15%) from KH2PO4 were added to the basal diet to construct standard curves from which bioavailable P release could be calculated for the phytase treatments. In all cases, phytase supplementation levels were based on an assessment of phytase premix activity (i.e., P release from Na phytate at pH 5.5). Linear (P < 0.01) responses in tibia ash and weight gain resulted from iP supplementation in all assays. In the first chick trial, supplementation of 500 phytase units (FTU)/kg of ECP resulted in superior (P < 0.01) weight gain and tibia ash values compared with 500 FTU/kg of Natuphos. Results of the second chick trial revealed P-release values of 0.032 and 0.028% for 500 FTU/kg Natuphos and Ronozyme, respectively, and these were lower (P < 0.01) than the 0.125% P-release value for 500 FTU/kg of ECP. Tibia ash responded quadratically (P < 0.05) in response to graded levels of ECP up to 1,500 FTU/kg in the third chick trial. Combining Natuphos with either Ronozyme or ECP in Chick Trial 4 revealed no synergism between phytases with different initiation sites of P removal. The pig trial involved 10 individually fed weanling pigs per diet, and and phytase enzymes were supplemented to provide 400 FTU/kg in diets containing 0.60% Ca. Based on the linear regression of fibula ash on supplemental iP intake (r2 = 0.87), P-release values were 0.081% for Natuphos, 0.043% for Ronozyme, and 0.108% for ECP. These trials revealed an advantage of the E. coli phytase over the commercial phytases in young chicks.

Adding wheat middlings, microbial phytase, and citric acid to corn-soybean meal diets for growing pigs may replace inorganic phosphorus supplementation.

Three experiments were conducted with 96 growing Landrace x Yorkshire x Duroc crossbreds to determine the collective effectiveness of cereal phytase from wheat middlings, microbial phytase, and citric acid in improving phytate-P bioavailability in corn-soy diets. In Exp. 1, 40 gilts (7 wk old) were fed five diets for 8 wk. Diets 1, 2, and 3 were low-P, corn-soybean meal diets (CSB) + 0, .1, or .2% inorganic P (Pi) as calcium phosphate, respectively. Diet 4 was a similar corn-soy diet that included 15% wheat middlings (461 cereal phytase U/kg). Diet 5 was the CSB + microbial phytase (1,200 U/kg; Natuphos, BASF, Mount Olive, NJ). In Exp. 2, 16 barrows (8 wk old) were fed two diets for 6 wk. Diet 1 was the same as Diet 3 of Exp. 1 (.2% Pi). Diet 2 was Diet 4 of Exp. 1 + microbial phytase (300 U/kg). In Exp. 3, 40 barrows and gilts (6 wk old) were fed four diets for 6 wk. Diets 1 and 2 were the same as those in Exp. 2. Diet 3 was Diet 2 of Exp. 2 + 1.5% citric acid. Diet 4 was similar to Diet 3 but contained 10 instead of 15% wheat middlings. In Exp. 1, pigs fed the low-P, CSB (Diet 1) had lower (P < .05) ADG, ADFI, plasma Pi concentration, bone strength, and mobility score than pigs of the other four treatments. Measurements for pigs fed the 15% wheat middlings diet were not significantly different from those of pigs fed the CSB + .1% Pi or microbial phytase. In Exp. 2, ADG (P=.06) during wk 1 to 3 and gain:feed ratio (P < .02) and plasma Pi concentration (P < .005) during all weeks favored pigs fed the CSB + .2% Pi compared with the other diet including 15% wheat middlings. In Exp. 3, identical ADG during all weeks and similar plasma Pi concentrations at wk 4 and 6 were observed between pigs fed the two citric acid diets (Diets 3 and 4) and the CSB + .2% Pi (Diet 1). Pigs fed Diet 4 (10% wheat middlings) had even higher (P < .02) gain:feed ratio during wk 1 to 3 than those fed Diet 1. It seems feasible to completely replace calcium phosphate with 10 to 15% wheat middlings, 300 U microbial phytase/ kg, and 1.5% citric acid in the corn-soy diets for growing pigs.

Effects of combining three fungal phytases with a bacterial phytase on plasma phosphorus status of weanling pigs fed a corn-soy diet.

The objective of this study was to determine possible synergistic effects of supplementing one of three fungal phytases: Aspergillus fumitagus PhyA (AFP),A. niger PhyA (ANP), or Peniophora lyci phytase (PLP) with an Escherichia coli AppA phytase (EP) in diets for pigs. Three experiments, each lasting for 4 wk, were conducted with a total of 106 weanling pigs (5 wk old). The corn-soybean meal basal diet (BD) contained no supplemental inorganic P. In Exp. 1, 35 pigs (8.6 +/- 1.0 kg BW) were fed (as-fed basis) BD + AFP at 750 U/ kg of feed, BD + inorganic P (0.2% P), or BD + PLP at 500, 750, or 1,000 U/kg feed. Pigs fed BD + AFP or BD + 0.2% P had higher (P < 0.05) plasma inorganic P concentrations than those fed BD + PLP at the end of the trial (wk 4). In Exp. 2, 35 pigs (8.1 +/- 0.9 kg BW) were fed BD + AFP, EP, PLP, a 1:1 mix of AFP:EP, or a 1:1 mix of PLP:EP at 500 U/kg. Pigs fed the AFP:EP mixture had growth performance and plasma measures similar to those fed either enzyme alone. Pigs fed the PLP:EP mixture had lower (P < 0.05) plasma alkaline phosphatase activity than those fed BD + PLP. Pigs fed BD + PLP had lower (P < 0.05) plasma inorganic P concentrations than pigs fed BD + EP, and higher (P < 0.05) plasma alkaline phosphatase activity than all other groups at wk 4. In Exp. 3, 36 pigs (9.1 +/- 1.2 kg BW) were fed BD + ANP, EP, or a 1:1 mix of ANP:EP at 500 U/kg feed. Pigs fed the two enzymes together had lower (P < 0.05) plasma inorganic P concentration than those fed BD + EP and lower (P < 0.05) plasma alkaline phosphatase activity than pigs fed BD + ANP at wk 4. In conclusion, although the four phytases showed different effects on plasma P status of weanling pigs, there was no synergistic effect between any of the three fungal phytases and the bacterial phytase on the plasma measures or growth performance under the conditions of the present study.

Supplemental phytases of microbial and cereal sources improve dietary phytate phosphorus utilization by pigs from weaning through finishing.

This experiment was conducted to measure the nutritional and metabolic responses of pigs fed diets with continuous supplementation of microbial and cereal phytase from weaning to finishing, and to determine the feasibility of complete replacement of inorganic P addition by supplemental phytase in swine diets. Forty-eight Landrace x Hampshire x Meishan pigs were divided into four groups. In phase 1 (10 to 50 kg BW), pigs in Groups 1, 2, 3, and 4 were fed a low-P, corn-soybean meal basal diet (BD), the BD plus microbial phytase (A. ficuum) at 1,200 units/kg, the BD plus 10% wheat bran (230 units of cereal phytase/kg), and the BD + .24% inorganic P (calcium phosphate), respectively. In phase 2 (51 to 90 kg BW), these pigs were fed a similar BD or the BD plus 1,000 microbial phytase units/kg, 20% wheat bran, or .20% inorganic P, respectively. Repeated measures included growth performance, P, Ca, and N balance, metatarsal and metacarpal bone strength, serum concentration of inorganic P, Ca, and 1,25-dihydroxycholecalciferol, and serum alkaline phosphatase activity. Pigs fed the BD supplemented with microbial phytase and pigs fed the BD supplemented with inorganic P showed almost identical responses for all variables. Pigs fed the BD supplemented with cereal phytase also had responses for various measures that were similar to those of pigs fed microbial phytase or inorganic P, except for some differences in serum inorganic P concentrations and bone strength in phase 1. Because of improvements in apparent digestibility of dietary P and N, fecal excretion of these two nutrients was reduced by 31 to 62% (P < .05) in pigs fed the BD supplemented with phytase compared with pigs fed inorganic P. It is physiologically feasible and environmentally advantageous to replace inorganic P with microbial or cereal phytase in corn-soybean meal diets for this type of pig through the entire growing-finishing period.

Effects of dietary selenium and vitamin E concentrations on phospholipid hydroperoxide glutathione peroxidase expression in reproductive tissues of pubertal maturing male rats.

Phospholipid hydroperoxide glutathione peroxidase (PHGPX) is the second intracellular selenium (Se)-dependent glutathione peroxidase (GSH-Px) identified in mammals. Our objectives were to determine the effect of dietary vitamin E and Se levels on PHGPX activity expression in testis, epididymis, and seminal vesicles of pubertal maturing rats, and the relationship of PHGPX expression with testicular development and sperm quality. Forty Sprague-Dawley male weanling rats (21-d old), were initially fed for 3 wk a torula yeast basal diet (containing 0.05 mg Se/kg) supplemented with marginal levels of Se (0.1 mg/kg as Na2SeO3) and vitamin E (25 IU/kg as all-rac-alpha-tocopheryl acetate). Then, rats were fed the basal diets supplemented with 0 or 0.2 mg Se/kg and 0 or 100 IU vitamin E/kg diet during the 3-wk period of pubertal maturing. Compared with the Se-supplemented rats, those fed the Se-deficient diets retained 31, 88, 67, and 50% of Se-dependent GSH-Px activities in liver, testis, epididymis, and seminal vesicles, respectively. Testes and seminal vesicles had substantially higher (5- to 20-fold) PHGPX activity than liver. Dietary Se deficiency did not affect PHGPX activities in the reproductive tissues, but reduced PHGPX activity in liver by 28% (P < 0.0001). Dietary vitamin E supplementation did not affect PHGPX activity in liver, whereas it raised PHGPX activity in seminal vesicles by 43% (P < 0.005). Neither dietary vitamin E nor Se levels affected body weight gains, reproductive organ weights, or sperm counts and morphology. In conclusion, expression of PHGPX activity in testis and seminal vesicles was high and regulated by dietary Se and vitamin E differently from that in liver.

Efficacy of a phytase derived from Escherichia coli and expressed in yeast on phosphorus utilization and bone mineralization in turkey poults.

A slope-response bioassay was conducted with male turkey poults to determine the sparing effect of P, based on improvements in bone mineralization in turkey poults, from 10 to 21 d of age when diets were supplemented with a novel phytase. Reference diets for calculation of the sparing effect of P contained 0.47, 0.55, 0.70, and 0.79% nonphytate phosphorus (NPP). Diets with varying dosages of a swine, Escherichia coli-derived AppA2 phytase (ECP) expressed in Pichia pastoris yeast (0, 250, 500, 750, and 1,000 U/kg) were added to the 0.47% NPP diet and improvements in bone mineralization determined the sparing effect of P supplied from ECP. Two additional reference diets were included that contained 500 U/kg from one of two commercial phytases (PA and PB) derived from Aspergillus and Peniophora. At 500 U/kg diet the ECP spared an additional 0.22% NPP (if calculated from tibia ash %), 0.18% NPP (if calculated from toe ash %), 0.24% NPP (if calculated from mg tibia ash), or 0.21% NPP (if calculated from mg toe ash). Phosphorus retention results validate bioassay results, in that 500 U ECP/kg resulted in 68.2% P being retained (0.49% of diet P retained) as compared with only 58.9% P being retained from the unsupplemented control diet (0.421% of diet P retained; P < 0.05).