J. W. Spears

Role of antioxidants and trace elements in health and immunity of transition dairy cows.

A number of antioxidants and trace minerals have important roles in immune function and may affect health in transition dairy cows. Vitamin E and beta-carotene are important cellular antioxidants. Selenium (Se) is involved in the antioxidant system via its role in the enzyme glutathione peroxidase. Inadequate dietary vitamin E or Se decreases neutrophil function during the perpariturient period. Supplementation of vitamin E and/or Se has reduced the incidence of mastitis and retained placenta, and reduced duration of clinical symptoms of mastitis in some experiments. Research has indicated that beta-carotene supplementation may enhance immunity and reduce the incidence of retained placenta and metritis in dairy cows. Marginal copper deficiency resulted in reduced neutrophil killing and decreased interferon production by mononuclear cells. Copper supplementation of a diet marginal in copper reduced the peak clinical response during experimental Escherichia coli mastitis. Limited research indicated that chromium supplementation during the transition period may increase immunity and reduce the incidence of retained placenta.

Micronutrients and immune function in cattle

Complex inter-relationships exist between certain micronutrients, immune function and disease resistance in cattle. Several micronutrients have been shown to influence immune responses. The relationship between deficiencies of some micronutrients and disease resistance is less clear. A number of studies have indicated that Cr supplementation may improve cell-mediated and humoral immune response as well as resistance to respiratory infections in stressed cattle. With respiratory-disease challenge models Cr generally does not affect disease resistance. Deficiencies of Cu, Se, vitamin E and Co in cattle reduce the ability of isolated neutrophils to kill yeast and/or bacteria. Cu deficiency reduces antibody production, but cell-mediated immunity is generally not altered. However, Cu deficiency appears to reduce production of interferon and tumour necrosis factor by mononuclear cells. Numerous studies have linked low vitamin E and/or Se status to increased susceptibility of dairy cows to intramammary infections. In contrast to findings in laboratory animals, marginal Zn deficiency does not appear to impair antibody production or lymphocyte responsiveness to mitogen stimulation in ruminants. Co deficiency has been associated with reduced resistance to parasitic infections. It is well documented that vitamin A-deficient animals are more susceptible to various types of infections. beta-Carotene, possibly via its antioxidant properties, may affect immune function and disease resistance independent of its role as a precursor of vitamin A.

Organic trace minerals in ruminant nutrition

This paper defines the various types (complexes, amino acid chelates and proteinates) of organic trace minerals commercially available and reviews research regarding their bioavailability and effects on physiological responses in ruminants. The addition of one or more organic trace minerals to cattle diets has increased growth, milk production, reproduction- and/or immune response in some studies. It is not possible to determine from some of the studies if the responses observed were due to the organic mineral(s) per se or simply to increased dietary mineral intake. Zinc methionine has been studied to the greatest extent of any of the chelated or metal complex products available. The mode of action of trace mineral chelates or complexes is largely unknown. Based on apparent absorption or tissue and blood concentrations, little evidence is available to suggest that organic trace minerals are considerably better absorbed than inorganic forms. In explaining the beneficial responses to certain organic trace minerals, the quantity of mineral absorbed may not be as important as the form of the mineral absorbed. Further studies with organic trace minerals are needed to better define conditions where performance or health responses may be expected and to determine the mode of action whereby organic trace mineral supplements improve ruminant performance.

Ctr1 Is an Apical Copper Transporter in Mammalian Intestinal Epithelial Cells in Vivo That Is Controlled at the Level of Protein Stability

Copper is an essential trace element that functions in a diverse array of biochemical processes that include mitochondrial respiration, neurotransmitter biogenesis, connective tissue maturation, and reactive oxygen chemistry. The Ctr1 protein is a high-affinity Cu+ importer that is structurally and functionally conserved in yeast, plants, fruit flies, and humans and that, in all of these organisms, is localized to the plasma membrane and intracellular vesicles. Although intestinal epithelial cell-specific deletion of Ctr1 in mice demonstrated a critical role for Ctr1 in dietary copper absorption, some controversy exists over the localization of Ctr1 in intestinal epithelial cells in vivo. In this work, we assess the localization of Ctr1 in intestinal epithelial cells through two independent mechanisms. Using immunohistochemistry, we demonstrate that Ctr1 localizes to the apical membrane in intestinal epithelial cells of the mouse, rat, and pig. Moreover, biotinylation of intestinal luminal proteins from mice fed a control or a copper-deficient diet showed elevated levels of both total and apical membrane Ctr1 protein in response to transient dietary copper limitation. Experiments in cultured HEK293T cells demonstrated that alterations in the levels of the glycosylated form of Ctr1 in response to copper availability were a time-dependent, copper-specific posttranslational response. Taken together, these results demonstrate apical localization of Ctr1 in intestinal epithelia across three mammalian species and suggest that increased Ctr1 apical localization in response to dietary copper limitation may represent an adaptive response to homeostatically modulate Ctr1 availability at the site of intestinal copper absorption.

Iron Transporters Are Differentially Regulated by Dietary Iron, and Modifications Are Associated with Changes in Manganese Metabolism in Young Pigs

To investigate the effects of dietary iron (Fe) on manganese (Mn) metabolism, 24 weaned pigs (21 d old) were blocked by litter and weight and randomly assigned to the following treatments: 1) no supplemental Fe [low Fe (L-Fe)]; 2) 100 mg supplemental Fe/kg [adequate Fe (A-Fe)]; and 3) 500 mg supplemental Fe/kg [high Fe (H-Fe)]. The basal diet was analyzed to contain 20 mg Fe/kg. Tissues were harvested after 32 d of feeding. Daily gain (least square means +/- SEM) was greater in A-Fe pigs (328.3 +/- 29.9 g/d) than in L-Fe pigs (224.0 +/- 11.2 g/d). Hemoglobin concentrations on d 32 were lower in L-Fe pigs (62 +/- 3.5 g/L) than in A-Fe pigs (128 +/- 5.6 g/L) and did not differ between pigs fed A-Fe and H-Fe (133 +/- 12.0 g/L). Liver Fe increased with increasing dietary Fe. Relative hepatic hepcidin expression was greater in pigs fed A-Fe and H-Fe than in those fed L-Fe. Relative expressions of duodenal divalent metal transporter 1 (DMT1) and solute carrier family 39 member 14 (ZIP14) were increased in L-Fe pigs compared with H-Fe pigs. Liver copper (Cu) was higher in L-Fe (0.56 +/- 0.04 mmol/kg) and H-Fe (0.58 +/- 0.04 mmol/kg) pigs than in A-Fe pigs (0.40 +/- 0.04 mmol/kg). Liver Mn was lower in H-Fe pigs (0.15 +/- 0.01 mmol/kg) than in A-Fe (0.23 +/- 0.02 mmol/kg) or L-Fe pigs (0.20 +/- 0.02 mmol/kg). Duodenal Mn concentrations were greater in L-Fe pigs than in A-Fe or H-Fe pigs. Fe deficiency in pigs increased gene expression of duodenal metal transporters (DMT1 and ZIP14) and supplementation with H-Fe reduced expression of DMT1 and ZIP14, which may have decreased absorption of Mn.

High dietary iron reduces transporters involved in iron and manganese metabolism and increases intestinal permeability in calves

A 56-d experiment was designed to examine the effect of high dietary Fe on metal transporters involved in Fe and Mn metabolism. Fourteen weaned Holstein calves were stratified by weight and randomly assigned to 1 of 2 treatments: 1) no supplemental Fe (normal Fe) or 2) 750mg of supplemental Fe/kg of dry matter (high Fe). Jugular blood was collected on d 0, 35, and 56. At the end of the trial, 6 calves per treatment were humanely killed and duodenal scrapings, liver, and heart were collected for analysis. Additionally, proximal duodenum was mounted on Ussing chambers to assess intestinal barrier integrity. Calves receiving high dietary Fe displayed decreased transepithelial resistance and increased apical-to-basolateral flux of radiolabeled mannitol, suggesting that high Fe created increased intestinal permeability. Feeding calves a diet high in Fe decreased average daily gain, dry matter intake, and feed efficiency. Hemoglobin and serum Fe concentrations did not differ due to dietary treatment. High dietary Fe increased concentrations of Fe in the liver, but did not affect heart or duodenal Fe concentrations. Duodenal Mn concentrations were lowered by feeding a high Fe diet, but liver and heart Mn concentrations were not affected. As determined by real-time reverse transcription PCR, relative hepatic expression of the gene that encodes the Fe regulatory hormone hepcidin was 5-fold greater in calves fed high dietary Fe. Hepcidin is released in response to increased Fe status and binds to the Fe export protein ferroportin causing ferroportin to be degraded, thereby reducing dietary Fe absorption. Confirmation of this result was achieved through Western blotting of duodenal protein, which revealed that ferroportin was decreased in calves fed high dietary Fe. Duodenal protein expression of divalent metal transporter 1 (DMT1), a Fe import protein that can also transport Mn, tended to be reduced by high dietary Fe. Transcript levels of several genes involved in Fe metabolism in liver and duodenum were unchanged by treatment. In summary, feeding calves a diet high in Fe induced a signal cascade (hepcidin) designed to reduce absorption of Fe (via reduced protein expression of ferroportin and DMT1) in a manner similar to that reported in rodents. Additionally, reduced levels of DMT1 protein appeared to decrease duodenal Mn, suggesting that Mn may also be a substrate for DMT1 in cattle.

Iron methionine as a source of iron for the neonatal pig

Neonatal pigs (143) were used to determine if adequate iron (Fe) would be stored following a single oral dose of Fe methionine (FeMet) to prevent anemia through 21 d of age. Treatments consisted of: control (no Fe), 200 mg injectable Fe as gleptoferron within 12 h of birth (at birth), 100 or 200 mg Fe as FeMet orally at birth, or 200 mg Fe as FeMet or FeSO4 orally on d 3. Pigs given injectable Fe had higher hemoglobin concentrations than other treatments at 14 and 21 d. Utilization of Fe from FeMet was greatest when given at 3 d compared to at birth. Based on hemoglobin concentrations at 21 d, relative bioavailability of Fe from FeMet given on d 3 was 180% compared to FeSO4. Iron methionine given orally was a safe, effective source of Fe, but a single oral dose of FeMet was not equivalent to injectable Fe.

Chromium propionate enhances insulin sensitivity in growing cattle

Thirty-six Angus and Angus×Simmental heifers, averaging 291 kg, were used to determine the effects of dietary Cr, in the form of Cr propionate (Cr Prop), on glucose metabolism and serum insulin concentrations following glucose administration. Heifers were stratified by body weight (BW) within a breed and randomly assigned to treatments. Treatments consisted of 0, 3, 6, or 9 mg of supplemental Cr/d from Cr Prop. Based on dry matter (DM) intakes, the daily doses of Cr were equivalent to 0.47, 0.94, and 1.42 mg of supplemental Cr/kg of DM. Heifers were individually fed a corn silage-based diet at a level of 2% of BW. Each heifer was also fed 0.45 kg of a ground corn supplement daily that served as a carrier for supplemental Cr. Glucose tolerance tests were performed on d 44 of the study. Glucose was infused via jugular catheters at a level of 0.45 g/kg of BW(0.75) over a course of 1 to 2 min. Blood samples were collected at -10, 0, 5, 10, 15, 30, 45, 60, 90, 120, 150, and 180 min relative to glucose dosing for glucose and insulin determination. Area under the glucose response curve was lower (1,603 vs. 1,964 mg/dL per minute) in heifers supplemented with Cr from 0 to 45 min following glucose challenge. Serum insulin concentrations were lower in Cr-supplemented heifers than in controls following glucose infusion. The molar ratio of insulin to glucose was also lower in Cr-supplemented heifers relative to controls. Serum insulin and serum insulin to glucose ratios did not differ among heifers supplemented with 3, 6, or 9 mg of Cr/d. Results indicate that Cr Prop supplementation increased tissue sensitivity to insulin in growing heifers. Based on insulin sensitivity, Cr requirements (as Cr Prop) of growing heifers can be met by supplementing with 3 mg of Cr/d or 0.47 mg of Cr/kg of DM.

Bioaccessibility of iron from soil is increased by silage fermentation

High dietary Fe can negatively affect absorption of other minerals and cause tissue damage through the production of free radicals. Cattle are often exposed to high dietary Fe, and soil ingestion may represent a major dietary source of Fe. Iron in soil is often found in the ferric form bound in insoluble complexes; however, exposure to an acidic environment similar to that occurring during silage fermentation may cause this Fe to be reduced to the more soluble ferrous form. To test this theory, a 2 x 2 x 3 factorial arrangement examining time, level, and type of soil addition to greenchop was used. Factors included 2 times of soil addition (before or after ensiling), 2 levels of soil inclusion (1 and 5% contamination, wet basis) and 3 types of soil (Cecil clay loam, 3.4% Fe; Georgeville silt loam, 4.3% Fe; and Dyke clay loam, 6.9% Fe). In addition, greenchop with no soil added was ensiled to serve as a control. Fresh corn greenchop was mixed with the appropriate type and level of soil and tightly packed in experimental silos. Fermentation was allowed to proceed for 90 d before silos were opened and silage was freeze-dried and ground. To simulate contamination after ensiling, each soil type was added to control silage at the 2 levels of inclusion. Addition of soil to greenchop before ensiling resulted in greater amounts of water soluble Fe compared with soil addition after ensiling, suggesting that Fe-soil binding properties were altered by ensiling. To test the potential bioaccessibility of Fe during ruminant digestion, an enzymatic in vitro system was modified to simulate ruminal, abomasal, and intestinal digestion. The presence of soil, regardless of time of addition, type, or inclusion level, resulted in greater soluble or bioaccessible Fe concentrations after all 3 phases when compared with control silage. Ensiling further increased soluble Fe concentrations after each phase when compared with silage contaminated with soil after ensiling. In addition, dialyzable Fe concentration (15,000 Da molecular weight cut off) following intestinal phase simulation was greater due to ensiling. Iron that becomes soluble during the intestinal phase may be available to the animal for absorption, and ensiling resulted in increased concentrations of potentially bioavailable Fe. These results suggest that soil contamination of harvested feeds before ensiling may represent a major source of bioavailable Fe in the diets of cattle.

Tolerance of cattle to increased dietary sulfur and effect of dietary cation-anion balance

The objective of this study was to determine if dietary cation-anion balance (DCAB) affects the concentration of S that can be tolerated by growing and finishing cattle without adversely affecting performance. Angus cross and Bradford steers (n=114; average initial BW=252.8 kg) were blocked by BW and breed, and randomly assigned within a block to treatment. The design was a 3 × 2 factorial arrangement of treatments with S (from NH(4)SO(4)) supplemented at 0, 0.15, or 0.30% of DM, and NaHCO(3) added at 0 or 1.0% of DM. Each treatment consisted of 3 pens containing 5 steers and 1 pen containing 4 steers. Steers were used in an 84-d growing study followed by a finishing study. A corn silage-based diet was fed during the growing study and a corn-based diet was fed during the finishing study. Steers were not randomized between experiments. The analyzed concentrations of S in the growing diets were 0.12, 0.30, and 0.46%, whereas the analyzed concentrations of S in the finishing diets were 0.13, 0.31, and 0.46% for treatments supplemented with 0, 0.15, and 0.30% S, respectively. Increasing DCAB by approximately 15 mEq/100 g of DM, by the addition of NaHCO(3,) did not affect (P > 0.36) performance during the growing or finishing studies. During the growing study DMI was not affected (P=0.29) by dietary S. Steers fed diets containing 0.30% S had greater ADG (P=0.02) and G:F (P=0.01) than those receiving 0.46% S, but similar (P > 0.36) performance to steers fed 0.12% S. During the finishing study, steers fed diets containing 0.46% S had less ADG than steers fed 0.13 (P=0.004) or 0.31% S (P=0.07), whereas ADG did not differ (P=0.18) among steers fed 0.13 and 0.31% S. Steers fed diets containing 0.31 (P=0.01) or 0.46% S (P=0.001) had less DMI than controls, but G:F was not affected (P=0.52) by S during the finishing study. Carcass characteristics did not differ (P > 0.18) among steers fed diets containing 0.13 and 0.31% S. Steers receiving diets containing 0.46% S had decreased HCW (P=0.001), quality (P=0.02), and yield grades (P=0.04) than steers receiving 0.13% S. Plasma Cu concentrations on d 101 of the finishing phase and liver Cu concentrations at slaughter were greater (P ≤ 0.05) in control steers compared with those fed diets containing 0.31 or 0.46% S. This study indicates that steers fed growing diets can tolerate up to 0.46% S with minimum effects on performance. Finishing steers tolerated diets containing 0.31% S without adverse affects on ADG or G:F. However, 0.46% S greatly decreased ADG and DMI, and increasing DCAB did not prevent these depressions.