A calcitonin receptor (CALCR) single nucleotide polymorphism is associated with growth performance and bone integrity in response to dietary phosphorus deficiency.

Although concerns over the environmental impact of excess P in the excreta from pig production and governmental regulations have driven research toward reducing dietary supplementation of P to swine diets for over a decade, recent dramatic increases in feed costs have further motivated researchers to identify means to further reduce dietary P supplementation. We have demonstrated that genetic background impacts P utilization in young pigs and have identified genetic polymorphisms in several target genes related to mineral utilization. In this study, we examined the impact of a SNP in the calcitonin receptor gene (CALCR) on P utilization in growing pigs. In Exp. 1, 36 gilts representing the 3 genotypes identified by this CALCR SNP (11, 12, and 22) were fed a P-adequate (PA) or a marginally P-deficient (approximately 20% less available P; PD) diet for 14 wk. As expected, P deficiency reduced plasma P concentration, bone strength, and mineral content (P < 0.05). However, the dietary P deficiency was mild enough to not affect the growth performance of these pigs. A genotype x dietary P interaction (P < 0.05) was observed in measures of bone integrity and mineral content, with the greatest reduction in bone strength and mineral content due to dietary P deficiency being associated with the allele 1. In Exp. 2, 168 pigs from a control line and low residual feed intake (RFI) line were genotyped for the CALCR SNP and fed a PA diet. As expected, pigs from the low RFI line consumed less feed but also gained less BW when compared with the control line (P < 0.05). Although ADFI did not differ between genotypes, pigs having the 11 genotype gained less BW (P < 0.05) than pigs having the 12 or 22 genotypes. Pigs of the 11 and 12 genotypes had bones that tolerated greater load when compared with animals having the 22 genotype (P < 0.05). A similar trend was observed in bone modulus and ash % (P < 0.10). These data are supportive of the association of this CALCR SNP with bone integrity and its response to dietary P restriction. Although the allele 1 is associated with greater bone integrity and mineral content during adequate P nutrition, it is also associated with the greatest loss in bone integrity and mineral content in response to dietary P restriction. Understanding the underlying genetic mechanisms that regulate P utilization may lead to novel strategies to produce more environmentally friendly pigs.

Response to dietary phosphorus deficiency is affected by genetic background in growing pigs.

Concern over the environmental effect of P excretion from pig production has led to reduced dietary P supplementation. To examine how genetics influence P utilization, 94 gilts sired by 2 genetic lines (PIC337 and PIC280) were housed individually and fed either a P-adequate diet (PA) or a 20% P-deficient diet (PD) for 14 wk. Initially and monthly, blood samples were collected and BW recorded after an overnight fast. Growth performance and plasma indicators of P status were determined monthly. At the end of the trial, carcass traits, meat quality, bone strength, and ash percentage were determined. Pigs fed the PD diet had decreased (P < 0.05) plasma P concentrations and poorer G:F (P < 0.05) over the length of the trial. After 4 wk on trial, pigs fed the PD diet had increased (P < 0.05) plasma 1,25(OH)(2)D(3) and decreased (P < 0.05) plasma parathyroid hormone compared with those fed the PA diet. At the end of the trial, pigs fed the PD diet had decreased (P < 0.05) BW, HCW, and percentage fat-free lean and tended to have decreased LM area (P = 0.06) and marbling (P = 0.09) and greater (P = 0.12) 10th-rib backfat than pigs fed the PA diet. Additionally, animals fed the PD diet had weaker bones and also decreased (P < 0.05) ash percentage and increased (P < 0.05) concentrations of 1alpha-hydroxylase and parathyroid hormone receptor mRNA in kidney tissue. Regardless of dietary treatment, PIC337-sired pigs consumed more feed and gained more BW than their PIC280-sired counterparts (P < 0.05) during the study. The PIC337-sired pigs also had greater (P < 0.05) HCW, larger (P < 0.01) LM area, and tended to have (P = 0.07) greater dressing percentage. Meat from the PIC337-sired pigs also tended to have greater (P = 0.12) concentrations of lactate but decreased (P = 0.07) concentrations of total glucose units 24 h postslaughter. Although plasma 1,25(OH)(2)D(3) concentrations were elevated (P < 0.05) in all the animals fed the PD diet, this elevation due to P deficiency tended (P = 0.09) to be greater in the PIC337-sired pigs after 12 wk on the treatment. The PIC337-sired pigs had stronger (P < 0.01) bones with greater ash percentage than the PIC280-sired pigs. The difference in the strength of the radii between the PIC337-sired pigs fed the PA and PD diets was greater than their PIC280-sired counterparts, which resulted in sire line x treatment interactions (P < 0.05). These data indicate differing mechanisms of P utilization between these genetic lines. Elucidating these mechanisms may lead to strategies to increase efficiency of growth in a more environmentally friendly manner.

The correlation of chemical and physical corn kernel traits with production performance in broiler chickens and laying hens.

A study was conducted to determine the influence on broiler chicken growth and laying hen performance of chemical and physical traits of corn kernels from different hybrids. A total of 720 male 1-d-old Ross-308 broiler chicks were allotted to floor pens in 2 replicated experiments with a randomized complete block design. A total of 240 fifty-two-week-old Hy-Line W-36 laying hens were allotted to cages in a randomized complete block design. Corn-soybean meal diets were formulated for 3 broiler growth phases and one 14-wk-long laying hen phase to be marginally deficient in Lys and TSAA to allow for the detection of differences or correlations attributable to corn kernel chemical or physical traits. The broiler chicken diets were also marginally deficient in Ca and nonphytate P. Within a phase, corn- and soybean-based diets containing equal amounts of 1 of 6 different corn hybrids were formulated. The corn hybrids were selected to vary widely in chemical and physical traits. Feed consumption and BW were recorded for broiler chickens every 2 wk from 0 to 6 wk of age. Egg production was recorded daily, and feed consumption and egg weights were recorded weekly for laying hens between 53 and 67 wk of age. Physical and chemical composition of kernels was correlated with performance measures by multivariate ANOVA. Chemical and physical kernel traits were weakly correlated with performance in broiler chickens from 0 to 2 wk of age (P<0.05, | r |<0.42). However, from 4 to 6 wk of age and 0 to 6 wk of age, only kernel chemical traits were correlated with broiler chicken performance (P<0.05, | r |<0.29). From 53 to 67 wk of age, correlations were observed between both kernel physical and chemical traits and laying hen performance (P<0.05, | r |<0.34). In both experiments, the correlations of performance measures with individual kernel chemical and physical traits for any single kernel trait were not large enough to base corn hybrid selection on for feeding poultry.

The correlation of chemical and physical corn kernel traits with growth performance and carcass characteristics in pigs.

Corn kernel composition may affect its nutritive value and, thus, pig growth performance and carcass characteristics. The objective of this study was to determine the influence of the chemical and physical traits of corn kernels from different hybrids on the growth performance and carcass characteristics of pigs. A total of 288 crossbred pigs were grown in a 3-phase program from 21 kg of BW until slaughter at 113 kg of BW with 12 pens (4 pigs/pen) per dietary treatment. Target BW for each phase were 20 to 40 kg (grower 1), 40 to 80 kg (grower 2), and 80 to 120 kg (finisher). In each phase, diets were formulated to be marginally deficient in Lys, TSAA, Ca, Na, and nonphytate P to improve the likelihood of detecting differences in performance due to corn hybrid. Each of 6 corn hybrids represented a wide range of kernel chemical and physical traits and was substituted for corn in a common diet formulation on an equal weight basis to make the 6 dietary treatments. Physical and chemical composition of the kernels were analyzed and correlated with performance measures by multivariate ANOVA. Kernel density was correlated with i.m. fat (IMF) content in LM (r = -.35, P < 0.05). Stenvert grinding time was correlated (P < 0.05) with ADG during the grower 1 phase (r = 0.26), ADFI during the grower 2 phase (r = 0.27), final BW (r = 0.27), and IMF (r = -0.36). The amylose content of the cornstarch was correlated (P < 0.05) with ADG during the grower 2 phase (r = -0.28) and with BW at the end of the grower 2 phase (r = -0.27). The NDF content of the kernels was correlated (P < 0.05) with ADG during the finisher phase (r = -0.30), final BW (r = -0.33), and number of days to market (r = 0.31). The ADF content of the kernels was correlated (P < 0.05) with ADG during the grower 1 phase (r = -0.26), final BW (r = -0.26), and IMF (r = 0.31). The correlations of performance measure variation with individual kernel hybrid physical and chemical traits were statistically significant yet not large enough to base corn hybrid selection for feeding pigs on any single kernel chemical or physical trait.

Dietary inclusion of colicin e1 is effective in preventing postweaning diarrhea caused by F18-positive Escherichia coli in pigs.

With worldwide concern over the use of antibiotics in animal agriculture and their contribution to the spread of antibiotic resistance, alternatives to conventional antibiotics are needed. Previous research in our laboratories has shown that colicin E1 is effective against some Escherichia coli strains responsible for postweaning diarrhea (PWD) in vitro. In this study we examined the efficacy of the dietary inclusion of colicin E1 in preventing experimentally induced PWD caused by F18-positive enterotoxigenic E. coli in young pigs. Twenty-four weaned pigs (23 days of age), identified by genotyping to be susceptible to F18-positive E. coli infections, were individually housed and fed diets containing 0, 11, or 16.5 mg colicin E1/kg diet. Two days after the start of the trial, all animals were orally inoculated with 1 x 10(9) CFU of each of two F18-positive E. coli strains isolated from pigs with PWD. The dietary inclusion of colicin E1 decreased the incidence and severity of PWD caused by F18-positive enterotoxigenic E. coli and improved the growth performance of the piglets. Additionally, the reduced incidence of PWD due to dietary colicin E1, lowered the levels of expression of the genes for interleukin 1beta and tumor necrosis factor beta in ileal tissues from these animals. The dietary inclusion of colicin E1 may be an effective alternative to conventional antibiotics in the diets of weaning pigs for the prevention of PWD caused by F18-positive enterotoxigenic E. coli.

Effect of dietary phosphorus and its interaction with genetic background on global gene expression in porcine muscle.

Environmental concerns and costs associated with dietary phosphorus (P) supplementation have lead to attempts to minimize the amount of P added to swine diets. In addition to its requirement for bone growth, dietary P is also necessary for muscular growth. To examine the effects of genetic background and dietary P on global gene expression in the muscle of young pigs, we utilized muscle tissue from 36 gilts sired from two different sire lines. These animals were fed either a P adequate, P deficient or P repletion diets for 14 days and showed differences in growth performance and bone integrity in response to the interaction of genetic background and dietary P. Total RNA from the loin muscle of these animals was obtained for microarray analysis. Significant differences (p<0.01) in gene expression were seen based on the effect of sire line (339 genes), dietary P (18 genes) and the interaction between sire line and dietary P (31 genes). The microarray data were validated by semi-quantitative real-time PCR. These results support our hypothesis that genetic background and dietary P treatment can affect the homeorhetic control of P metabolism in pigs. Genes identified as differentially expressed in this study may be excellent candidate genes for additional work to elucidate genotype specific P requirements as well as to identify a genetic background that can maintain superior growth in a more environmentally friendly manner.

SNP discovery, expression and association analysis for the SDHD gene in pigs.

The SDHD gene was examined for single nucleotide polymorphisms (SNP) as well as for expression changes in the Longissimus dorsi muscle of commercial pigs with different potential for growth. Three SNPs, including one previously described in the coding region and two new ones in the 3'-UTR, were found. The normalized expression of SDHD was correlated with growth, meat quality and sensory traits (p<0.05). For the commercial pigs used in this study, as well as a Berkshire x Yorkshire resource population, the SNPs have been associated (p<0.05) with: growth, carcass composition, meat quality and sensory traits. Despite the fact that the described SNPs were not significantly associated with the normalized expression values, the SDHD SNPs and expression were associated with growth and meat quality traits in pigs.

SNP discovery, expression and association analysis for the SDHD gene in pigs.

The SDHD gene was examined for single nucleotide polymorphisms (SNP) as well as for expression changes in the Longissimus dorsi muscle of commercial pigs with different potential for growth. Three SNPs, including one previously described in the coding region and two new ones in the 3'-UTR, were found. The normalized expression of SDHD was correlated with growth, meat quality and sensory traits (p < 0.05). For the commercial pigs used in this study, as well as a Berkshire x Yorkshire resource population, the SNPs have been associated (p < 0.05) with: growth, carcass composition, meat quality and sensory traits. Despite the fact that the described SNPs were not significantly associated with the normalized expression values, the SDHD SNPs and expression were associated with growth and meat quality traits in pigs.

Gene expression profiling: insights into skeletal muscle growth and development.

Microarray technology is now available for many livestock species, and animal scientists are beginning to utilize the technology to address issues of importance to animal agriculture. This review discusses how microarray technology has been applied to study global gene expression changes in skeletal muscle. For example, microarrays have been used to elucidate gene function in knockout mice, evaluate breed differences, evaluate the effects of hormone administration, and evaluate the effects of diet. Data generated from these global gene expression studies are providing new insights to stimulate future hypothesis-driven research.

Reflections on uncertainty in risk assessment and risk management by the Society of Environmental Toxicology and Chemistry (SETAC) precautionary principle workgroup.

Quantitative uncertainty assessments and the distribution of risk are under scrutiny and significant criticism has been made of null hypothesis testing when careful consideration of Type I (false positive) and II (false negative) error rates have not been taken into account. An alternative method, equivalence testing, is discussed yielding more transparency and potentially more precaution in the quantifiable uncertainty assessments. With thousands of chemicals needing regulation in the near future and low public trust in the regulatory process, decision models are required with transparency and learning processes to manage this task. Adaptive, iterative, and learning decision making tools and processes can help decision makers evaluate the significance of Type I or Type II errors on decision alternatives and can reduce the risk of committing Type III errors (accurate answers to the wrong questions). Simplistic cost-benefit based decision-making tools do not incorporate the complex interconnectedness characterizing environmental risks, nor do they enhance learning, participation, or include social values and ambiguity. Hence, better decision-making tools are required, and MIRA is an attempt to include some of the critical aspects.

Colicin concentrations inhibit growth of Escherichia coli O157:H7 in vitro.

Escherichia coli O157:H7 is a virulent foodborne pathogen that causes severe human illness and inhabits the intestinal tract of food animals. Colicins are antimicrobial proteins produced by E. coli strains that inhibit or kill other E. coli. In the present Study, the efficacy of three pore-forming colicins (El, N, and A) were quantified in vitro against E. coli O157:H7 strains 86-24 and 933. Colicins E1 and N reduced the growth of E. coli O157:H7 strains, but the efficacy of each colicin varied among strains. Colicin E1 was more effective against both strains of E. coli O157:H7 than colicins A and N and reduced (P < 0.05) populations of E. coli O157:H7 at concentrations <0.1 microg/ml. These potent antimicrobial proteins may potentially provide an effective and environmentally sound preharvest strategy to reduce E. coli O157:H7 in food animals.

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.

Biotechnological development of effective phytases for mineral nutrition and environmental protection.

Phytases are hydrolytic enzymes that initiate the release of phosphate from phytate (myo-inositol hexakisphosphate), the major phosphorus (P) form in animal feeds of plant origin. These enzymes can be supplemented in diets for food animals to improve P nutrition and to reduce P pollution of animal excreta. This mini-review provides a synopsis of the concept of "ideal phytase" and the biotechnological approaches for developing such an enzyme. Examples of Escherichia coli AppA and Aspergillus fumigatus PhyA are presented to illustrate how new phytases are identified from microorganisms and developed by genetic engineering based on the gene sequences and protein structures of these enzymes. We also discuss the characteristics of different heterologous phytase expression systems, including those of plants, bacteria, fungi, and yeast.

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.

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.

Dietary intrinsic phytate protects colon from lipid peroxidation in pigs with a moderately high dietary iron intake.

High iron consumption has been proposed to relate to an increase in the risk of colon cancer, whereas high levels of supplemental sodium phytate effectively reduce iron-induced oxidative injury and reverse iron-dependent augmentation of colorectal tumorigenesis. However, the protective role of intrinsic dietary phytate has not been determined. In this study, we examined the impact of removing phytate present in a corn-soy diet by supplemental microbial phytase on susceptibility of pigs to the oxidative stress caused by a moderately high dietary iron intake. Thirty-two weanling pigs were fed the corn-soy diets containing two levels of iron (as ferrous sulfate, 80 or 750 mg/kg diet) and microbial phytase (as Natuphos, BASF, Mt. Olive, NJ, 0 or 1200 units/kg). Pigs fed the phytase-supplemented diets did not receive any inorganic phosphorus to ensure adequate degradation of phytate. After 4 months of feeding, liver, colon, and colon mucosal scrapings were collected from four pigs in each of the four dietary groups. Colonic lipid peroxidation, measured as thiobarbituric acid reacting substances (TBARS), was increased by both the high iron (P< 0.0008) and phytase (P< 0.04) supplementation. Both TBARS and F2-isoprostanes, an in vivo marker of lipid peroxidation, in colonic mucosa were affected by dietary levels of iron (P< 0.03). Mean hepatic TBARS in pigs fed the phytase-supplemented, high iron diet was 43%-65% higher than that of other groups although the differences were nonsignificant. Moderately high dietary iron induced hepatic glutathione peroxidase activity (P= 0.06) and protein expression, but decreased catalase (P< 0.05) in the colonic mucosa. In conclusion, intrinsic phytate in corn and soy was protective against lipid peroxidation in the colon associated with a moderately high level of dietary iron.

Mixed-function oxidase studies in the redfish, Sciaenops ocellata, from Galveston Bay, Texas.

The metabolism of parathion to para-nitrophenol (PNP) in redfish (Sciaenops ocellata) liver microsomes has been both identified and characterized. This mixed-function oxidase (MFO) reaction in redfish requires NADPH and is inhibited by carbon monoxide. It exhibits a temperature optimum of 25 degrees C but no clear pH optimum between 7.0-8.5. Redfish hepatic microsomal MFOs were not induced by 2.5 days after a single i.p. injection of 88 mg Aroclor 1254/kg body wt, but under the same dosage and time conditions male albino Swiss mice were significantly induced (p less than 0.05).