Testing of an oral dosing technique for double-crested cormorants, Phalacocorax auritus, laughing gulls, Leucophaeus atricilla, homing pigeons, Columba livia, and western sandpipers, Calidris mauri, with artificially weather MC252 oil.

Scoping studies were designed to determine if double-crested cormorants (Phalacocorax auritus), laughing gulls (Leucophaues atricilla), homing pigeons (Columba livia) and western sandpipers (Calidris mauri) that were gavaged with a mixture of artificially weathered MC252 oil and food for either a single day or 4-5 consecutive days showed signs of oil toxicity. Where volume allowed, samples were collected for hematology, plasma protein electrophoresis, clinical chemistry and electrolytes, oxidative stress and organ weigh changes. Double-crested cormorants, laughing gulls and western sandpipers all excreted oil within 30min of dose, while pigeons regurgitated within less than one hour of dosing. There were species differences in the effectiveness of the dosing technique, with double-crested cormorants having the greatest number of responsive endpoints at the completion of the trial. Statistically significant changes in packed cell volume, white cell counts, alkaline phosphatase, alanine aminotransferase, creatine phosphokinase, gamma glutamyl transferase, uric acid, chloride, sodium, potassium, calcium, total glutathione, glutathione disulfide, reduced glutathione, spleen and liver weights were measured in double-crested cormorants. Homing pigeons had statistically significant changes in creatine phosphokinase, total glutathione, glutathione disulfide, reduced glutathione and Trolox equivalents. Laughing gulls exhibited statistically significant decreases in spleen and kidney weight, and no changes were observed in any measurement endpoints tested in western sandpipers.

Toxic effects of orally ingested oil from the Deepwater Horizon spill on laughing gulls.

The explosion of the Deepwater Horizon oil rig released, millions of gallons of oil into the environment, subsequently exposing wildlife, including numerous bird species. To determine the effects of MC252 oil to species relevant to the Gulf of Mexico, studies were done examining multiple exposure scenarios and doses. In this study, laughing gulls (Leucophaeus atricilla, LAGU) were offered fish injected with MC252 oil at target doses of 5 or 10mL/kg bw per day. Dosing continued for 27 days. Of the adult, mixed-sex LAGUs used in the present study, ten of 20 oil exposed LAGUs survived to the end of the study; a total of 10 of the oil exposed LAGUs died or were euthanized within 20 days of initiation of the study. Endpoints associated with oxidative stress, hepatic total glutathione (tGSH), oxidized glutathione (GSSG) and reduced glutathione (rGSH) significantly increased as mean dose of oil increased, while the rGSH:GSSG ratio showed a non-significant negative trend with oil dose. A significant increase in 3-methyl histidine was found in oil exposed birds when compared to controls indicative of muscle wastage and may have been associated with the gross observation of diminished structural integrity in cardiac tissue. Consistent with previous oil dosing studies in birds, significant changes in liver, spleen, and kidney weight when normalized to body weight were observed. These studies indicate that mortality in response to oil dosing is relatively common and the mortality exhibited by the gulls is consistent with previous studies examining oil toxicity. Whether survival effects in the gull study were associated with weight loss, physiologic effects of oil toxicity, or a behavioral response that led the birds to reject the dosed fish is unknown.

Dermal exposure to weathered MC252 crude oil results in echocardiographically identifiable systolic myocardial dysfunction in double-crested cormorants (Phalacrocorax auritus).

During the Deepwater Horizon Natural Resource Damage Assessment, gross morphologic cardiac abnormalities, including softer, more distensible musculature, were noted upon gross necropsy in hearts from laughing gulls and double-crested cormorants exposed to weathered MC252 crude oil. A species specific, echocardiographic technique was developed for antemortem evaluation of function that was used to evaluate and better characterize cardiac dysfunction. Control (n=12) and treated (n=13) cormorant groups of similar sex-ratio and ages were dermally treated with approximately 13ml of water or weathered MC252 crude oil, respectively, every 3 days for 6 dosages. This resulted in a low to moderate external exposure. Upon visualization and clinical assessment of the hearts of all test subjects, comprehensive diagnostic cardiographic measurements were taken twice, prior to oil application and after a 21day dermal oil exposure. Oil-treated birds showed a decrease in cardiac systolic function, as characterized by an increased left ventricular internal dimension-systole and left ventricular stroke volume as well as concurrent decreased left ventricular ejection fraction and left ventricular fractional shortening when compared to both control birds' and the treated birds' time zero values. These changes are indicative of a possible dilative cardiomyopathy induced by oil exposure, although further elucidation of possible collagen damage is recommended. Arrhythmias including tachycardia in two treated birds and bradycardia in all treated birds were documented, indicating further clinically significant abnormalities induced by MC252 oil that warrant further investigation. A statistically significant increase in free calcium concentration, important to muscular and neurologic function in treated birds was also noted. This study documents that weathered MC252 oil caused clinically significant cardiac dysfunction that could result in mortality and decrease recruitment.

Reprint of: Overview of avian toxicity studies for the Deepwater Horizon Natural Resource Damage Assessment.

The Oil Pollution Act of 1990 establishes liability for injuries to natural resources because of the release or threat of release of oil. Assessment of injury to natural resources resulting from an oil spill and development and implementation of a plan for the restoration, rehabilitation, replacement or acquisition of natural resources to compensate for those injuries is accomplished through the Natural Resource Damage Assessment (NRDA) process. The NRDA process began within a week of the Deepwater Horizon oil spill, which occurred on April 20, 2010. During the spill, more than 8500 dead and impaired birds representing at least 93 avian species were collected. In addition, there were more than 3500 birds observed to be visibly oiled. While information in the literature at the time helped to identify some of the effects of oil on birds, it was not sufficient to fully characterize the nature and extent of the injuries to the thousands of live oiled birds, or to quantify those injuries in terms of effects on bird viability. As a result, the US Fish and Wildlife Service proposed various assessment activities to inform NRDA injury determination and quantification analyses associated with the Deepwater Horizon oil spill, including avian toxicity studies. The goal of these studies was to evaluate the effects of oral exposure to 1-20ml of artificially weathered Mississippi Canyon 252 oil kg bw-1 day-1 from one to 28 days or one to five applications of oil to 20% of the bird's surface area. It was thought that these exposure levels would not result in immediate or short-term mortality but might result in physiological effects that ultimately could affect avian survival, reproduction and health. These studies included oral dosing studies, an external dosing study, metabolic and flight performance studies and field-based flight studies. Results of these studies indicated changes in hematologic endpoints including formation of Heinz bodies and changes in cell counts. There were also effects on multiple organ systems, cardiac function and oxidative status. External oiling affected flight patterns and time spent during flight tasks indicating that migration may be affected by short-term repeated exposure to oil. Feather damage also resulted in increased heat loss and energetic demands. The papers in this special issue indicate that the combined effects of oil toxicity and feather effects in avian species, even in the case of relatively light oiling, can significantly affect the overall health of birds.

Overview of avian toxicity studies for the Deepwater Horizon Natural Resource Damage Assessment.

The Oil Pollution Act of 1990 establishes liability for injuries to natural resources because of the release or threat of release of oil. Assessment of injury to natural resources resulting from an oil spill and development and implementation of a plan for the restoration, rehabilitation, replacement or acquisition of natural resources to compensate for those injuries is accomplished through the Natural Resource Damage Assessment (NRDA) process. The NRDA process began within a week of the Deepwater Horizon oil spill, which occurred on April 20, 2010. During the spill, more than 8500 dead and impaired birds representing at least 93 avian species were collected. In addition, there were more than 3500 birds observed to be visibly oiled. While information in the literature at the time helped to identify some of the effects of oil on birds, it was not sufficient to fully characterize the nature and extent of the injuries to the thousands of live oiled birds, or to quantify those injuries in terms of effects on bird viability. As a result, the US Fish and Wildlife Service proposed various assessment activities to inform NRDA injury determination and quantification analyses associated with the Deepwater Horizon oil spill, including avian toxicity studies. The goal of these studies was to evaluate the effects of oral exposure to 1-20ml of artificially weathered Mississippi Canyon 252 oil kg bw-1 day-1 from one to 28 days or one to five applications of oil to 20% of the bird's surface area. It was thought that these exposure levels would not result in immediate or short-term mortality but might result in physiological effects that ultimately could affect avian survival, reproduction and health. These studies included oral dosing studies, an external dosing study, metabolic and flight performance studies and field-based flight studies. Results of these studies indicated changes in hematologic endpoints including formation of Heinz bodies and changes in cell counts. There were also effects on multiple organ systems, cardiac function and oxidative status. External oiling affected flight patterns and time spent during flight tasks indicating that migration may be affected by short-term repeated exposure to oil. Feather damage also resulted in increased heat loss and energetic demands. The papers in this special issue indicate that the combined effects of oil toxicity and feather effects in avian species, even in the case of relatively light oiling, can significantly affect the overall health of birds.

Effects of phytase supplementation in mature horses fed alfalfa hay and pelleted concentrate diets.

An experiment was conducted to study P digestibility in mature horses because of the growing environmental concerns regarding P runoff and previous equine research focused mostly on young and growing animals or used ponies as a model. Phytase supplementation of swine and poultry diets can result in greater phytate-P digestibility, leading to a decreased need for inorganic P supplementation and a decrease in P excreted to the environment; this, however, has not been demonstrated in the horse. Six mature Arabian geldings were fed 6 diets consisting of pelleted concentrate and alfalfa hay. The concentrates consisted mainly of soybean hulls, ground corn, wheat midds, broken rice, and beet pulp, and phytase was added to the concentrates accordingly before pelleting. There were 3 diet types: control (concentrate and hay), high P (greater P concentrate and hay), and forage only, and each diet type included 1 phytase-supplemented diet and 1 non-phytase-supplemented diet, resulting in 6 total diets. Phytase supplementation for the forage only diet was accomplished by feeding a nominal amount of concentrate formulated solely as a vehicle for the phytase. Horses had unrestricted access to water throughout the experiment. Using a Latin square design, all horses received all diets over a period of 12 wk. In each week, the new diet was fed for 11 d of diet acclimation, which was followed by a 3-d total collection of feces and urine for each horse. There was no effect (P < 0.05) of phytase supplementation on P output in the urine or feces, resulting in no differences in P apparent digestibility. Analysis of the feed and feces for phytate revealed a 93% average disappearance rate of phytate, indicating that horses are highly capable of degrading phytate and that phytase supplementation was not beneficial. Thus, the results indicate that mature horses are able to maintain a near 0 P balance, with adequate P provided in the diet even as phytate, and increased P intakes above requirement may result in increased potentially detrimental outputs to the environment.

Effect of dietary organic microminerals on starter pig performance, tissue mineral concentrations, and liver and plasma enzyme activities.

Weanling pigs (n = 160) were used to evaluate dietary essential microminerals (Cu, Fe, Mn, Se, and Zn) on performance, tissue minerals, and liver and plasma enzymatic activities during a 35-d postweaning period. A randomized complete block design with 5 treatments and 8 replicates was used in this study. Organic microminerals were added to complex nursery diets at 0 (basal), 50, 100, or 150% of the requirements of microminerals listed by the 1998 NRC. A fifth treatment contained inorganic microminerals at 100% NRC and served as the positive control. Pigs were bled at intervals with hemoglobin (Hb), hematocrit (Hct), glutathione peroxidase, and ceruloplasmin activities determined. Six pigs at weaning and 1 pig per pen at d 35 were killed, and the liver, heart, loin, kidney, pancreas, and the frontal lobe of the brain were collected for micromineral analysis. The liver was frozen in liquid N for determination of enzymatic activities. The analyzed innate microminerals in the basal diet met the NRC requirement for Cu and Mn but not Fe, Se, and Zn. Performance was not affected from 0 to 10 d postweaning, but when microminerals were added to diets, ADG, ADFI, and G:F improved (P < 0.01) from 10 to 35 d and for the overall 35-d period. Pigs fed the basal diet exhibited parakeratosis-like skin lesions, whereas those fed the supplemental microminerals did not. This skin condition was corrected after a diet with the added microminerals was fed. When the basal diet was fed, Hb and Hct declined, but supplemental microminerals increased Hb and Hct values. Liver catalase activity increased (P < 0.01) when microminerals were fed. The Mn superoxide dismutase activity tended to decline quadratically (P = 0.06) when supplemental microminerals were fed above that of the basal diet. Liver plasma glutathione peroxidase activities were greater (P < 0.01) when dietary organic and inorganic micromineral were fed. Liver concentrations of microminerals increased linearly (P < 0.01) as dietary microminerals increased, indicating that the liver was the primary storage organ. Micromineral tissue concentrations were least in pigs fed the basal diet and increased (quadratic, P < 0.01) to the 50% level of organic microminerals in the various tissues collected. The results indicated that innate microminerals, Cu and Mn, from a complex nursery diet may meet the micromineral needs of the weaned pig, but the need for Fe, Se, or Zn was not met by the basal diet.

Transporters in the absorption and utilization of zinc and copper.

Before the discovery and elucidation of transporters, mammals were thought to cotransport Cu or Zn as an anionic complex, such as binding with an AA as a chelate or a receptor such as transferrin. In 1995, the first mammalian Zn transporter (ZnT) gene, ZnT1, was identified. However, 2 protein families are now thought to be involved in Zn transport. The ZnT family reduces intracellular Zn by aiding in efflux from the cell or promoting the influx into intracellular vesicles. The mechanism of ZnT transport against a Zn concentration gradient is unknown; however, only ZnT1 appears to be located at the plasma membrane. It has been shown to respond in tissues in a variety of ways to Zn reduction and supplementation. In our laboratory, we have found ZnT1 and metallothionein to work in concert during pharmacological Zn supplementation. The second protein family, Zip proteins, provides Zn transport from extracellular fluid or intracellular vesicles into the cytoplasm and has not been identified in a livestock species. Like Zn, no good indicator of status has been identified for Cu. However, the recent identification of Cu transporters and chaperones gives researchers the opportunity to understand the regulation of Cu trafficking where the proteins are modified by posttranslational mechanisms. Two Cu transporters, Ctr1 and Ctr3, mediate high-affinity Cu uptake. A small cytoplasmic protein, MURR1, has been identified in human hepatic tissue, but its role in Cu metabolism is unknown. The discovery of Cu chaperones that are involved in facilitating Cu absorption into proteins may provide an excellent status indicator. It has been shown that the Cu chaperone for Cu/Zn superoxide dismutase (CCS) is increased in tissue of Cu-deficient rats, induced when moderately high Zn diets are fed. We have recently found CCS in the young pig. Other Cu chaperone proteins that have been identified are COX17 and Atox1. As with CCS, they are involved in making Cu available to apo-enzymes inside the cell. It is essential that these new molecular findings be used to evaluate the bioavailability of and nutritional need for Cu and Zn in livestock.

Utilization of distillers dried grains with solubles and phytase in sow lactation diets to meet the phosphorus requirement of the sow and reduce fecal phosphorus concentration.

Two experiments were completed to determine the potential for using distillers dried grains with solubles (DDGS) in diets with or without phytase to provide available P, energy, and protein to highly productive lactating sows without increasing their fecal P. In Exp. 1, the dietary treatments were as follows: (1) corn and soybean meal with 5% beet pulp (BP) or (2) corn and soybean meal with 15% DDGS (DDGS). Besides containing similar amounts of fiber, diets were isonitrogenous (21% CP, 1.2% Lys) and isophosphorus (0.8% P). Sixty-one sows were allotted to dietary treatments at approximately 110 d of gestation (when they were placed in farrowing crates) based on genetics, parity, and date of farrowing. Sows were gradually transitioned to their lactation diet. On d 2 of lactation, litters were cross-fostered to achieve 11 pigs/litter. Sows and litters were weighed on d 2 and 18. Fecal grab samples were collected on d 7, 14, and 18 of lactation. Dietary treatment did not affect the number of pigs weaned (10.9 vs. 10.8) or litter weaning weight. On d 14, DDGS sows had less fecal P concentration than BP sows (28.3 vs. 32.8 mg/g; P = 0.04). Fecal Ca of sows fed DDGS decreased for d 7, 14, and 18 (55.6, 51.4, and 47.1 mg/g of DM, respectively; P = 0.05) but not for BP sows. In Exp. 2, the dietary treatments were as follows: (1) corn and soybean meal (CON), (2) CON + 500 phytase units of Natuphos/kg diet, as fed (CON + PHY), (3) corn and soybean meal with 15% DDGS and no phytase (DDGS), or (4) DDGS + 500 FTU of Natuphos/kg of diet, as fed (DDGS + PHY). Sows (n = 87) were managed as described for Exp 1. Litter BW gain (46.0, 46.3, 42.1, and 42.2 kg; P = 0.25) and sow BW loss (8.1, 7.2, 7.4, and 6.3 kg for CON, CON + PHY, DDGS, and DDGS + PHY, respectively; P = 0.97) were not affected by dietary treatment. Fecal P concentration did not differ among dietary treatments but was reduced at d 14 and 18 compared with d 7 (P = 0.001). However, fecal phytate P concentration was decreased by the addition of DDGS when DDGS and DDGS + PHY were compared with the CON sows except on d 7 (P < 0.05). Sows fed CON diet had greater fecal phytate P than sows fed DDGS, and sows fed DDGS + PHY had less fecal phytate P than sows fed DDGS with no phytase (P = 0.001). Although these experiments were only carried out for 1 lactation, these results indicate that highly productive sows can sustain lactation performance with reduced fecal phytate P when fed DDGS and phytase in lactation diets.

Impact of supplement withdrawal and wheat middling inclusion on bone metabolism, bone strength, and the incidence of bone fractures occurring at slaughter in pigs.

The objective of this study was to determine if supplement withdrawal (omission of dietary vitamin and trace mineral premixes and 2/3 of inorganic P) 28-d preslaughter and the feeding of wheat middlings (dietary concentrations of 5, 15, or 30% from weaning to 16 kg, 16 to 28 kg, and 28 kg to slaughter, respectively) affect bone metabolism, bone strength, bone density, and the incidence of bone fractures at slaughter in pigs. Crossbred barrows (n = 64) were assigned to a 2 x 2 factorial arrangement of treatments (with or without supplement withdrawal, and with or without wheat middlings). Serum was collected on d 0, 14, and 27 of the preslaughter withdrawal period to determine changes in the concentrations of osteocalcin, an indicator of bone formation, and pyridinoline, an indicator of bone resorption. The serum osteocalcin and pyridinoline concentrations on d 14 and 27 were analyzed as change from the d-0 concentration. At slaughter, radiographs of the lumbar vertebrae and of the right and left femurs were taken to determine the incidence of bone fractures. Third metacarpal bones were analyzed for bone mineral density, peak load, ultimate shear stress, and percent ash. Supplement withdrawal increased (P < 0.05) serum osteocalcin and pyridinoline concentrations, indicating an increase in osteoblast activity and bone resorption. Supplement withdrawal decreased (P < 0.01) bone mineral density, peak load, ultimate shear stress, and percent ash of the metacarpal bones. Dietary wheat middling inclusion did not alter bone quality. Neither supplement withdrawal nor wheat middling inclusion affected the incidence of bone fractures at slaughter. The results of this study indicate that removing inorganic P, vitamin premix, and trace mineral premix for 28 d preslaughter increases bone turnover and decreases bone quality.

Effects of dietary zinc and iron supplementation on mineral excretion, body composition, and mineral status of nursery pigs.

Two experiments were conducted to evaluate the effects of dietary Zn and Fe supplementation on mineral excretion, body composition, and mineral status of nursery pigs. In Exp. 1 (n = 24; 6.5 kg; 16 to 20 d of age) and 2 (n = 24; 7.2 kg; 19 to 21 d of age), littermate crossbred barrows were weaned and allotted randomly by BW, within litter, to dietary treatments and housed individually in stainless steel pens. In Exp. 1, Phases 1 (d 0 to 7) and 2 (d 7 to 14) diets (as-fed basis) were: 1) NC (negative control, no added Zn source); 2) ZnO (NC + 2,000 mg/kg as Zn oxide); and 3) ZnM (NC + 2,000 mg/kg as Zn Met). In Exp. 2, diets for each phase (Phase 1 = d 0 to 7; Phase 2 = d 7 to 21; Phase 3 = d 21 to 35) were the basal diet supplemented with 0, 25, 50, 100, and 150 mg/kg Fe (as-fed basis) as ferrous sulfate. Orts, feces, and urine were collected daily in Exp. 1; whereas pigs had a 4-d adjustment period followed by a 3-d total collection period (Period 1 = d 5 to 7; Period 2 = d 12 to 14; Period 3 = d 26 to 28) during each phase in Exp. 2. Blood samples were obtained from pigs on d 0, 7, and 14 in Exp. 1 and d 0, 7, 21, and 35 in Exp. 2 to determine hemoglobin (Hb), hematocrit (Hct), and plasma Cu, (PCu), Fe (PFe), and Zn (PZn). Pigs in Exp. 1 were killed at d 14 (mean BW = 8.7 kg) to determine whole-body, liver, and kidney mineral concentrations. There were no differences in growth performance in Exp. 1 or 2. In Exp. 1, pigs fed ZnO or ZnM diets had greater (P < 0.001) dietary Zn intake during the 14-d study and greater fecal Zn excretion during Phase 2 compared with pigs fed the NC diet. Pigs fed 2,000 mg/kg, regardless of Zn source, had greater (P < 0.010) PZn on d 7 and 14 than pigs fed the NC diet. Whole-body Zn, liver Fe and Zn, and kidney Cu concentrations were greater (P < 0.010), whereas kidney Fe and Zn concentrations were less (P < 0.010) in pigs fed pharmacological Zn diets than pigs fed the NC diet. In Exp. 2, dietary Fe supplementation tended to increase (linear, P = 0.075) dietary DMI, resulting in a linear increase (P < 0.050) in dietary Fe, Cu, Mg, Mn, P, and Zn intake. Subsequently, a linear increase (P < 0.010) in fecal Fe and Zn excretion was observed. Increasing dietary Fe resulted in a linear increase in Hb, Hct, and PFe on d 21 (P < 0.050) and 35 (P < 0.010). Results suggest that dietary Zn or Fe additions increase mineral status of nursery pigs. Once tissue mineral stores are loaded, dietary minerals in excess of the body's requirement are excreted.

Effects of iron supplementation on binding activity of iron regulatory proteins and the subsequent effect on growth performance and indices of hematological and mineral status of young pigs.

Two experiments were conducted to evaluate the effects of supplemental Fe on the binding activity of iron regulatory proteins (IRP) and the subsequent effect on growth performance and indices of hematological and mineral status of young pigs. In Exp. 1, male pigs (n = 10; 1.8 kg; age = 14 +/- 1 h) were allotted by BW to two treatments (five pigs per treatment). Treatments administered by i.m. injection were as follows: 1) 1 mL of sterile saline solution (Sal); and 2) 1 mL of 200 mg Fe as Fe-dextran (Fe). Pigs were bled (d 0 and 13) to determine hemoglobin (Hb), hematocrit (Hct), transferrin (Tf), and plasma Fe (PFe), and then killed (d 13) to determine spontaneous and 2-mercaptoethanol (2-ME)-inducible IRP RNA binding activity in liver and liver and whole-body mineral concentrations. Contemporary pigs (n = 5; 2.2 kg; age = 14 +/- 2 h) were killed at d 0 to establish baseline (BL1) measurements. In Exp. 2, pigs (six pigs per treatment; 6.5 kg; age = 19 +/- 3 d) were fed a basal diet (Phase 1 = d 0 to 7; Phase 2 = d 7 to 21; Phase 3 = d 21 to 35) supplemented with 0 or 150 mg/kg of Fe as ferrous sulfate and killed at d 35 (18.3 kg; age = 54 +/- 3 d). In addition, pigs (n = 5; 5.9 kg; age = 19 +/- 3 d) were killed at the start of Exp. 2 to establish baseline (BL2) measurements, and liver samples were collected and analyzed for IRP RNA binding activity. In Exp. 1, no difference (P = 0.482) was observed in ADG. On d 13, Fe-treated pigs had greater (P = 0.001) Hb, Hct, and PFe and less (P = 0.002) Tf than Sal-treated pigs. Whole-body Fe concentration was greater (P = 0.002) in Fe- vs. Sal-treated pigs. Treated pigs (Fe or Sal) had greater (P = 0.006) whole-body Cu and less (P = 0.002) whole-body Ca, Mg, Mn, P, and Zn concentrations than BL1. Liver Fe concentration was greater (P = 0.001) in Fe- vs. Sal-treated pigs, but liver Fe concentration of Sal-treated pigs was less (P = 0.001) than that of BL1 pigs. Sal-treated pigs had greater (P = 0.004) spontaneous IRP binding activity than Fe-treated pigs. In Exp. 2, spontaneous and 2-ME inducible IRP binding activities were greater (P = 0.013 and 0.005, respectively) in pigs fed diets containing 0 vs. 150 mg of added Fe/kg of diet. Moreover, pigs fed either treatment for 35 d had greater (P = 0.001) 2-ME inducible IRP binding activity than BL2 pigs. Results indicate that IRP binding activity is influenced by Fe supplementation. Subsequently, other indicators of Fe status are affected via the role of IRP in posttranscriptional expression of Fe storage and transport proteins.

Dietary pharmacological or excess zinc and phytase effects on tissue mineral concentrations, metallothionein, and apparent mineral retention in the newly weaned pig.

Feeding pharmacological zinc (Zn) to weaned pigs improves growth, and dietary phytase improves P and Zn availability. Metallothionein (MT) increases in the duodenum, kidney, and liver of pigs fed 1000 mg Zn/kg with phytase or 2000 mg Zn/kg with or without phytase when fed for 14 d postweaning. The goal of this study was to determine the effects of feeding pharmacological Zn and phytase on tissue minerals, MT, mineral excretion, and apparent retention. Twenty-four newly weaned pigs (20 d; 7.2 kg) were individually fed twice daily, a basal diet supplemented with 0, 1000, or 4000 mg Zn/kg as Zn oxide, without or with phytase (500 phytase units [FTU]/kg) for 14 d, followed by a basal diet (100 mg Zn/kg) without phytase for 7 d. Pigs fed 4000 mg Zn/kg without phytase had higher (p=0.01) plasma, hepatic, renal Zn, renal Cu, and hepatic, renal, and jejunal MT than pigs fed the basal diet or 1000 mg Zn/kg. Duodenal MT was higher (p=0.0001) in pigs fed 1000 and 4000 mg Zn/kg than in pigs fed the basal diet. In pigs fed 1000 and 4000 mg Zn/kg, Zn loading occurred during the first 11 d of supplementation; by d 14, excess Zn was being excreted in the feces.

Effects of dietary iron supplementation on growth performance, hematological status, and whole-body mineral concentrations of nursery pigs.

An experiment was conducted to evaluate the effects of supplementing increasing concentrations of Fe to the diet of nursery pigs on growth performance and indices of hematological and mineral status. Pigs (n = 225; 6.5 kg; 19 +/- 3 d) were allotted randomly by BW, litter, and gender to one of five dietary treatments (five pigs per pen; nine pens per treatment). Basal diets for each phase (Phase 1: d 0 to 7; Phase 2: d 7 to 21; Phase 3: d 21 to 35) were formulated to contain minimal Fe concentration and then supplemented with 0, 25, 50, 100, and 150 mg Fe/kg of diet (as-fed basis) from ferrous sulfate. Three pigs per pen (n = 135) were chosen and bled throughout (d 0, 7, 21, and 35) to determine hemoglobin (Hb), hematocrit (Hct), transferrin (Tf), and plasma Fe (PFe). In addition, pigs (n = 5; 5.9 kg; 19 +/- 3 d) from the contemporary group were killed at d 0 to establish baseline (BL), and 30 pigs (six pigs/treatment) were killed at d 35 to determine whole-body and liver mineral concentrations. The improvements in growth performance during Phase 2 (ADG = linear, P = 0.04; ADFI = linear, P = 0.10; G:F = quadratic, P = 0.07) were of sufficient magnitude that dietary treatments tended to increase ADG (linear, P = 0.08), ADFI (quadratic, P = 0.09), and G:F (quadratic, P = 0.10) for the 35-d experiment. Hematological variables were not affected until d 21, at which time dietary Fe supplementation resulted in a linear increase (P = 0.03) in Hb, Hct, and PFe. This linear increase (P = 0.001) was maintained until d 35 of the experiment; however, dietary treatments resulted in a linear decrease (P = 0.01) in Tf on d 35. Whole-body Fe concentration increased (linear, P = 0.01) in pigs due to increasing dietary Fe concentrations. Moreover, pigs fed for 35 d had greater (P = 0.02) whole-body Fe, Zn, Mg, Mn, Ca, and P concentrations and lower (P = 0.001) whole-body Cu concentration than BL. Hepatic Fe concentration increased (linear, P = 0.001) in pigs due to dietary treatments; however, the hepatic Fe concentration of all pigs killed on d 35 was lower (P = 0.001) than the BL. Results suggest that Fe contributed by feed ingredients was not sufficient to maintain indices of Fe status. The decrease in Fe stores of the pigs was not severe enough to reduce growth performance. Even so, the lessening of a pig's Fe stores during this rapid growth period may result in the occurrence of anemia during the subsequent grower and finisher periods.

Effect of Se on selenoprotein activity and thyroid hormone metabolism in beef and dairy cows and calves.

Although Se is essential for antioxidant and thyroid hormone function, factors influencing its requirement are not well understood. A survey and two experiments were conducted to determine the influence of cattle breed and age on selenoprotein activity and the effect of maternal Se supplementation on cow and calf selenoprotein activity and neonatal thyroid hormone production. In our survey, four cowherds of different ages representing three breeds were bled to determine the influence of breed and age on erythrocyte glutathione peroxidase activity (RBC GPX-1). All females were nonlactating, pregnant, and consumed total mixed diets (Holstein) or grazed pasture (Angus and Hereford). In our survey of beef breeds, yearlings had greater average RBC GPX-1 activity than mature cows. In Exp. 1, neonatal Holstein heifers (n = 8) were bled daily from 0 to 6 d of age to determine thyroid hormone profile. An injection of Se and vitamin E (BO-SE) was given after the initial bleeding. Thyroxine (T4) and triiodothyronine (T3) concentrations were greatest on d 0 and decreased (P < 0.05) continuously until d 5 postpartum (156.13 to 65.88 and 6.69 to 1.95 nmol/L, d 0 to 5 for T4 and T3, respectively). Reverse T3 concentrations were 3.1 nmol/L on d 0 and decreased (P < 0.05) to 0.52 nmol/ L by d 5. In Exp. 2, multiparous Hereford cows were drenched weekly with either a placebo containing 10 mL of double-deionized H2O (n = 14) or 20 mg of Se as sodium selenite (n = 13). After 2 mo of treatment, Se-drenched cows had greater (P < 0.01) plasma concentrations than control cows (84.92 vs. 67.08 ng/mL), and at parturition, they had plasma Se concentrations twofold greater than (P < 0.05) control cows (95.51 vs. 47.14 ng Se/mL). After 4 mo, cows receiving Se had greater (P < 0.05) RBC GPX-1 activity than controls; this trend continued until parturition. Colostrum Se concentration was twofold greater (P < 0.05) in Se-drenched cows than control cows (169.97 vs. 87.00 ng/mL). Calves born to cows drenched with Se had greater (P < 0.05) plasma Se concentration, RBC GPX-1, and plasma glutathione peroxidase activity on d 0 compared with calves born to control cows. By d 7, no differences in plasma glutathione peroxidase activity in calves were observed. Maternal Se supplementation did not influence calf thyroid hormone concentrations. Selenium provided by salt and forages is not adequate for cattle in Se-deficient states.

Impact of vitamin and mineral supplement withdrawal and wheat middling inclusion on finishing pig growth performance, fecal mineral concentration, carcass characteristics, and the nutrient content and oxidative stability of pork.

A study was conducted to determine if supplement withdrawal (omission of dietary vitamin and trace mineral premixes and a two-thirds reduction in dietary inorganic phosphorus) for 28 d preslaughter and the feeding of wheat middlings (dietary concentrations of 5, 15, and 30% from weaning to 16, 16 to 28, and 28 kg to slaughter, respectively) affect growth performance, carcass characteristics, and fecal mineral concentrations ofthe pig, as well as the nutrient content and oxidative stability of the longissimus dorsi muscle. Crossbred pigs (n = 64) were blocked by weight and assigned to one of four dietary treatments in a 2 x 2 factorial design (with or without supplement withdrawal, and with or without wheat middlings). Supplement withdrawal and wheat middling inclusion did not influence average daily gain (ADG), average daily feed intake, gain/feed, or carcass traits, except for a decrease (P < 0.01) in the ADG of pigs from 28 to 65 kg when fed wheat middlings. Supplement withdrawal decreased (P < 0.01) fecal Ca, P, Cu, Fe, Mn, and Zn concentrations. In diets containing full vitamin and mineral supplementation, wheat middling inclusion decreased (P < 0.01) fecal Ca, Cu, Fe, and Zn concentrations and increased (P < 0.01) fecal Mn. Supplement withdrawal decreased (P < 0.05) concentrations of riboflavin, niacin, and P in the longissimus dorsi muscle, but did not affect longissimus dorsi thiamin, vitamin E, Fe, Cu, Zn, and Ca concentrations. Inclusion of wheat middlings increased (P < 0.04) longissimus dorsi thiamin, niacin, riboflavin, and vitamin E concentrations and decreased (P < 0.04) Cu concentrations. However, wheat middling inclusion did not affect (P > 0.05) longissimus dorsi Ca, P, Fe, and Zn concentrations. Dietary treatment did not affect either Cu/Zn superoxide dismutase or glutathione peroxidase activity in the longissimus dorsi. The results from this study indicate that supplement withdrawal and dietary wheat middling inclusion alter pork nutrient content and fecal mineral concentration, but not the oxidative stability of pork.

Effect of vitamin E and selenium on iron utilization in neonatal pigs.

Supplying adequate iron (Fe) to neonatal pigs to support normal growth and hematological and antioxidant status, while preventing iron toxicity, is a challenge for producers. Three experiments were conducted to determine the effect of frequency and route of Fe administration with or without vitamin E (E) and selenium (Se) on growth, Fe, and antioxidant status of neonatal pigs. In Exp. 1, 12 pigs from dams with reduced E status were fed a semipurified diet without added Fe from d 3 to d 14 of age. At d 6 of age, pigs received the following i.m. injections: 1) FE, 1 mL containing 200 mg of Fe (iron dextran); 2) FEE, treatment FE plus 1 mL containing 300 IU of vitamin E (d-alpha tocopherol); or 3) FESEE, 1.03 mL containing 200 mg of Fe (iron dextran), .15 mg of Se (sodium selenite), and 15 IU of vitamin E (d-alpha tocopherol). Pigs were weighed daily and blood was collected at 3, 7, and 14 d of age. From d 8 to 14, growth was depressed (P < .05) in pigs injected with FESEE. At 14 d of age, pigs injected with FE or FEE had increased (P < .05) hemoglobin (Hb) concentration. Ceruloplasmin activity (CP) was greater (P < .05) at d 7 of age than at d 3 or 14 regardless of treatment. In Exp. 2, 3-d-old pigs (n = 94) received the following: 1) FE, 200 mg Fe (iron dextran) i.m.; (2) FEE, treatment FE plus 300 IU vitamin E i.m.; 3) EFE, 300 IU vitamin E i.m. followed by 200 mg Fe (iron dextran) i.m. 24 h later; or 4) OFE, 100 mg Fe and 10 mg Cu orally. On d 21 of age, one-half of the pigs in each treatment received a second dose of their respective treatment. Blood samples (n = 60) were obtained on d 3 and 21 of age. Pigs injected with FE, FEE, or EFE had greater (P < .05) Hb at d 21 than pigs given OFE. Copper/zinc superoxide dismutase (Cu/ZnSOD) activity was greater (P < .05) at d 21 with OFE than with the other treatments. At 65 d of age, ADG did not differ among treatments. In Exp. 3, pigs (n = 150, in three farrowing groups) were injected with 200 mg of Fe (iron dextran) on d 1 or d 1 and 14. Blood samples were obtained on d 7 and 21 of age. Hemoglobin concentration on d 21 was improved equally by both treatments. Catalase and Cu/ZnSOD activities were increased (P < .05) on d 21 of the experiment compared with d 7 regardless of treatment. Growth was not affected by injection frequency. Results from these experiments indicate that one Fe injection (200 mg) for pigs from sows fed adequate vitamin E will result in adequate growth and hemoglobin concentration with today's improved genetics.

Early- and traditionally weaned nursery pigs benefit from phase-feeding pharmacological concentrations of zinc oxide: effect on metallothionein and mineral concentrations.

Benefits of feeding pharmacological concentrations of zinc (Zn) provided by Zn oxide (ZnO) to 21-d conventionally weaned pigs in the nursery have been documented; however, several management questions remain. We conducted two experiments to evaluate the effect on growth from feeding 3,000 ppm Zn as ZnO during different weeks of the nursery period. In Exp. 1 (n = 138, 11.5 d of age, 3.8 kg BW) and Exp. 2 (n = 246, 24.5 d of age, 7.2 kg BW), pigs were fed either basal diets containing 100 ppm supplemental Zn (adequate) or the same diet with an additional 3,000 ppm Zn (high) supplied as ZnO. Pigs were fed four or two dietary phases in Exp. 1 and 2, respectively, that changed in dietary ingredients and nutrient content (lysine and crude protein) to meet the changing physiological needs of the pigs for the 28-d nursery period. Dietary Zn treatments were 1) adequate Zn fed wk 1 to 4, 2) high Zn fed wk 1, 3) high Zn fed wk 2, 4) high Zn fed wk 1 and 2, 5) high Zn fed wk 2 and 3, and 6) high Zn fed wk 1 to 4. In Exp. 1 and 2, pigs fed high Zn for wk 1 and 2 or the entire 28-d nursery period had the greatest (P < .05) ADG. During any week, pigs fed high Zn had greater concentrations of hepatic metallothionein and Zn in plasma, liver, and kidney than those pigs fed adequate Zn (P < .05). In summary, both early- and traditionally weaned pigs need to be fed pharmacological concentrations of Zn provided as ZnO for a minimum of 2 wk immediately after weaning to enhance growth.

Influence of age and magnesium on calcium metabolism in rats.

This study evaluates the effect of dietary magnesium concentration on calcium metabolism in rats of differing ages. Young (3 wk) and old (18 mo) Fischer 344 rats were fed the AIN-76A diet modified to contain either low (218 mg/kg) or adequate (419 mg/kg) Mg for 4 wk. Some rats subsequently underwent a metabolic balance study (12 d duration). Other rats were gavaged with approximately 220 KBq (6 microCi) of 47Ca; daily fecal and urine collections were made and periodic whole body radioactivity determined. Femurs were removed and analyzed. Calcium retention and balance were not affected by Mg in young rats. In old rats low Mg intake increased apparent Ca balance. Young rats retained about 3.25 times more of the original dose of 47Ca than did old rats. Young rats retained more 47Ca in the femur than did old rats; Mg intake had little effect. Aging accelerated Ca turnover rate, and whole body retention data suggest that adequate Mg does not significantly reduce Ca turnover.

Colonic impaction in guinea pigs fed a chemically defined diet: a case report.

Pathogen-free female Hartley guinea pigs were divided into four diet groups to determine optimal nitrogen requirements for growth, pregnancy and lactation. Three chemically defined test diets varying in total nitrogen were compared with commercial guinea pig feed. Within 13 days, 24 of 78 animals from the three test diet groups died or required euthanasia. Affected guinea pigs were depressed, lost weight and had distended abdomens. Distension, impaction and, in most cases, rupture of the spiral of the ascending colon and subsequent peritonitis were observed at necropsy. Husbandry changes which resulted in a decreased incidence of impactions included increased acclimatization time on commercial feed post-shipping, decreased animal density per cage and cessation of water medication with oxytetracycline.