Forms of selenium in vitamin-mineral mixes differentially affect serum alkaline phosphatase activity, and serum albumin and blood urea nitrogen concentrations, of steers grazing endophyte-infected tall fescue.

The goal of this study was to test the hypothesis that sodium selenite (ISe), SEL-PLEX (OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral mix would differentially affect serological and hepatic parameters of growing steers grazing toxic endophyte-infected tall fescue-mixed forage pasture. Predominately Angus steers (BW = 183 ± 34 kg) were randomly selected from herds of fall-calving cows grazing endophyte-infected tall fescue-mixed pasture and consuming vitamin-mineral mixes that contained 35 ppm Se as ISe, OSe, and MIX forms. Steers were weaned, depleted of Se for 98 d, and subjected to summer-long common grazing of an endophyte-infected tall fescue-mixed pasture (0.51 ppm total ergovaline + ergovalinine; 10.1 ha). Steers were assigned (n = 8 per treatment) to the same Se form treatments upon which they were raised. Se treatments were administered by daily top-dressing 85 g of vitamin-mineral mix onto 0.23 kg soyhulls, using in-pasture Calan gates. The PROC MIXED procedure of SAS was used to assess the effect of Se form treatments on serum parameters at day 0, 22, 43, 64, and 86. After slaughter, the effect of Se treatment on hepatic alkaline phosphatase (tissue nonspecific isoform, TNALP) mRNA, protein, and albumin protein content was assessed using the PROC GLM procedure of SAS. Fisher's protected LSD procedure was used to separate treatment means. Partial correlation analysis was used to evaluate the relationship among whole blood Se concentration and serum parameters, accounting for the effect of time. Across periods, MIX steers had more (P ≤ 0.04) serum albumin than OSe and ISe steers, respectively. However, the relative hepatic bovine serum albumin protein content was not affected (P = 0.28) by Se treatments. Serum alkaline phosphatase activity was greater (P ≤ 0.01) in MIX and OSe steers. Similarly, hepatic TNALP protein content in MIX steers was greater (P = 0.01) than ISe steers. Partial correlation analysis revealed that serum albumin, blood urea nitrogen, and alkaline phosphatase activity were correlated (r ≥ 0.23, P ≤ 0.02) with whole blood Se concentration. In summary, consumption of 3 mg Se/d as OSe or MIX forms of Se in vitamin-mineral mixes increased serum albumin concentration and alkaline phosphatase activity, the reduction of which is associated with fescue toxicosis. We conclude that the organic forms of Se ameliorated the depression of 2 of known serological biomarkers of fescue toxicosis.

Forms of selenium in vitamin-mineral mixes differentially affect the expression of genes responsible for prolactin, ACTH, and α-MSH synthesis and mitochondrial dysfunction in pituitaries of steers grazing endophyte-infected tall fescue.

The goal of this study was to test the hypothesis that sodium selenite (inorganic Se, ISe), SEL-PLEX (organic forms of Se, OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter pituitary transcriptome profiles in growing beef steers grazing an endophyte-infected tall fescue (E+) pasture. Predominately Angus steers (BW = 183 ± 34 kg) were randomly selected from fall-calving cows grazing E+ pasture and consuming VM mixes that contained 35 ppm Se as ISe, OSe, or MIX forms. Steers were weaned, depleted of Se for 98 d, and subjected to summer-long common grazing of a 10.1 ha E+ pasture containing 0.51 ppm ergot alkaloids. Steers were assigned (n = 8 per treatment) to the same Se-form treatments on which they were raised. Selenium treatments were administered by daily top-dressing 85 g of VM mix onto 0.23 kg soyhulls, using in-pasture Calan gates. As previously reported, serum prolactin was greater for MIX (52%) and OSe (59%) steers vs. ISe. Pituitaries were collected at slaughter and changes in global and selected mRNA expression patterns determined by microarray and real-time reverse transcription PCR analyses, respectively. The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The form of Se affected the expression of 542 annotated genes (P < 0.005). Integrated pathway analysis found a canonical pathway network between prolactin and pro-opiomelanocortin (POMC)/ACTH/α-melanocyte-stimulating hormone (α-MSH) synthesis-related proteins and that mitochondrial dysfunction was a top-affected canonical pathway. Targeted reverse transcription-PCR analysis found that the relative abundance of mRNA encoding prolactin and POMC/ACTH/α-MSH synthesis-related proteins was affected (P < 0.05) by the form of Se, as were (P ≤ 0.05) mitochondrial dysfunction-related proteins (CYB5A, FURIN, GPX4, and PSENEN). OSe steers appeared to have a greater prolactin synthesis capacity (more PRL mRNA) vs. ISe steers through decreased dopamine type two receptor signaling (more DRD2 mRNA), whereas MIX steers had a greater prolactin synthesis capacity (more PRL mRNA) and release potential by increasing thyrotropin-releasing hormone concentrations (less TRH receptor mRNA) than ISe steers. OSe steers also had a greater ACTH and α-MSH synthesis potential (more POMC, PCSK2, CPE, and PAM mRNA) than ISe steers. We conclude that form of Se in VM mixes altered expression of genes responsible for prolactin and POMC/ACTH/α-MSH synthesis, and mitochondrial function, in pituitaries of growing beef steers subjected to summer-long grazing an E+ pasture.

Glutathione content and expression of proteins involved with glutathione metabolism differs in longissimus dorsi, subcutaneous adipose, and liver tissues of finished vs. growing beef steers.

Increased tissue redox state may result in sub-optimal growth. Our goal was to determine if glutathione (GSH) content and expression of proteins involved with GSH metabolism change in longissimus dorsi (LD), subcutaneous adipose (SA), and liver tissues of growing vs. finishing steer phenotypes. Tissues were taken from 16 Angus steers (BW = 209 ± 29.4 kg) randomly assigned (n = 8) to develop through Growing (final BW = 301 ± 7.06 kg) vs. Finished (final BW = 576 ± 36.9 kg) growth phases, and at slaughter had achieved different rib-eye area (REA) (53.2, 76.8 cm2), marbling scores (296, 668), and 12th rib adipose thickness (0.54, 1.73 cm), respectively (Amino Acids, doi:10.1007/s00726-018-2540-8). GSH content (mg/g wet tissue) was determined by a commercial assay and the relative content of target protein and mRNA in tissue homogenates was determined by Western blot and reverse-transcribed PCR analyses, respectively. The effect of growth phase (Finished vs. Growing) was assessed by ANOVA using the GLM procedure of SAS. The LD of Finished steers had more (P < 0.04) GSH (42%) and GSH synthesizing (GCLC, 61%; GCLM, 21%) and metabolizing (GPX1, 42%; GPX3, 73%; GGT1, 56%) enzymes, and less (P < 0.02) GPX2 (46%), EAAC1 (30%) and glutamine synthetase (GS) (28%), whereas GTRAP3-18 and ARL6IP1 did not differ (P > 0.57). Principal component analysis found that GSH content of LD was associated with REA and marbling score. The SA of Finished steers had less (P < 0.04) GSH (38%), GSH metabolizing (GPX4, 52%; GGT1, 71%) enzyme mRNA, and GTRAP3-18 (123%) and ARL6IP1 (43%), whereas the mRNA content of GSH-synthesizing enzymes and content of EAAC1 and GS did not differ (P > 0.32). The liver of Finished steers had less (P < 0.02) mRNA content of GSH synthesizing (GCLC, 39%; GSS 29%) and metabolizing (GPX1, 30%) enzymes, and more (P < 0.01) GSTM1 metabolizing enzyme (114%). The change in GSH content as steers fattened indicate an increased antioxidant capacity in the LD of Finished steers, and a decreased antioxidant capacity in SA, consistent with changes in enzyme and transporter expression. Changes in liver enzyme and transporter expression were consistent with no change in GSH content. The relationship of EAAC1 regulatory proteins (GTRAP3-18, ARL6IP1) to GSH, EAAC1, and GS content differs and changes as Growing steers develop into Finished phenotypes. These findings provide mechanistic insight into how antioxidant capacity occurs in tissues of economic and metabolic importance as cattle fatten.

Form of supplemental selenium fed to cycling cows affects systemic concentrations of progesterone but not those of estradiol.

In areas where soils are deficient in selenium (Se), dietary supplementation of this trace mineral directly to cattle is recommended. Selenium status affects fertility, and the form of Se supplemented to cows affects tissue-specific gene expression profiles. The objective of this study was to determine whether the form of Se consumed by cows would affect follicular growth and the production of steroids. Thirty-three Angus-cross cows that had ad libitum access of a mineral mix containing 35 ppm of Se in free-choice vitamin-mineral mixes as either inorganic (ISe), organic (OSe), or a 50/50 mix of ISe and OSe (MIX) for 180 days were used. After 170 days of supplementation, all cows were injected with 25-mg PGF2α to induce regression of the CL and then monitored for behavioral estrus (Day 0). From Day 4 to Day 8 after estrus, follicular growth was determined by transrectal ultrasonography. On Day 6, cows were injected with PGF2α (20 then 15 mg, 8-12 hours apart) to induce regression of the developing CL and differentiation of the dominant follicle of the first follicular wave into a preovulatory follicle. On Day 8, 36 hours after PGF2α (20 mg), the contents of the preovulatory follicle were aspirated by ultrasound-guided follicular puncture. Blood collected on Days 6 and 8 and follicular fluid collected on Day 8 was analyzed for concentrations of progesterone and estradiol. Form of Se supplemented to cows affected (P = 0.04) the systemic concentration of progesterone on Day 6, but not on Day 8. Form of Se did not affect the systemic concentration of estradiol on Day 6 or Day 8. Form of Se tended to affect (P = 0.07) the concentration of progesterone, but not that of estradiol, in the follicular fluid. Form of Se did not affect diameter of the dominant ovarian follicle on Days 4 to 6, but tended to affect (P = 0.08) the diameter of the preovulatory follicle on Day 8. Our results suggest that form of Se fed to cows affects the production of progesterone but not that of estradiol. Further investigation of organic Se-induced increases in progesterone and potentially the effects of increased progesterone on the establishment of pregnancy, especially in cows of lower fertility, is warranted.

Individual intake of free-choice mineral mix by grazing beef cows may be less than typical formulation assumptions and form of selenium in mineral mix affects blood Se concentrations of cows and their suckling calves.

The objectives of this study were to determine (1) the individual ad libitum intake of mineral mix by beef cows managed under a year-long, fall-calving, forage-based production regimen and (2) if Se form in the mineral mix affected the blood Se concentrations of cows and suckling calves. Twenty-four late-gestation (6 to 8 months) Angus-cross cows (2.7 ± 0.8 years; body weight [BW] = 585 ± 58 kg) were blocked by BW and randomly assigned (n = 8) to a mineral supplement treatment (TRT) containing 35 ppm Se as either inorganic (ISe; sodium selenite), organic (OSe; Sel-Plex®), or a 1:1 combination of ISe/OSe (MIX). Cows commonly grazed a 10.1-ha predominately tall fescue pasture and had individual ad libitum access to TRT using in-pasture Calan gates. Cows calved from August to November and calves had common ad libitum access to creep feed and a mineral supplement that lacked Se. Cow jugular blood was taken at 28-day intervals (13 periods) and calf blood was taken with cows from birth through weaning. Individual cow mineral mix (mean = 54.0 ± 7.0 g/day, range = 97.3 to 27.9 ± 7.4 g/day) and Se (mean = 1.82 ± 0.25 mg/day, range = 3.31 to 0.95 ± 0.25 mg/day) intakes were affected by period (P < 0.0001), but not by cow Se TRT (P > 0.30). Cow blood Se (0.109 to 0.229 ± 0.01 µg/mL) was affected (P < 0.002) by period, Se form, and their interaction, with ISe < MIX for periods 8 and 11, ISe < OSe for all periods except period 1, and MIX < OSe for periods 2 to 4, 7, 8, 10, and 12. Calf blood Se (in micrograms Se per milliliter) was correlated with cow blood Se and affected (P < 0.0001) by cow Se TRT, with ISe (0.07 to 0.11) < MIX (0.10 to 0.15) = OSe (0.16 to 0.19). These data reveal that (1) mean supplemental ad libitum cow mineral intake was 36% less than the typical formulation intake expectations (85 g/day) and, correspondingly, mean supplemental Se intake was 33% less than that allowed by the FDA and (2) cow Se TRT differentially affected both cow and calf blood Se concentrations, resulting in adequate concentrations for all cows but inadequate concentrations for ISe calves.

Selenium content in blood fractions and liver of beef heifers is greater with a mix of inorganic/organic or organic versus inorganic supplemental selenium but the time required for maximal assimilation is tissue-specific.

Selenium (Se) content of feedstuffs is dependent on the Se level of the soil. Even though Se in grass and forage crops is primarily present in organic forms, Se is commonly supplemented in cattle diets in an inorganic (sodium selenite) form in geographic regions where Se soil concentrations are low. The purpose of this study was to answer two important questions about inorganic (ISe) vs organic (OSe) forms of dietary supplementation of Se (3 mg/day) to growing beef heifers (0.5 kg/day): (1) what would the effect of supplementing Se with an equal blend of ISe:OSe (Mix) have on Se tissue concentrations and (2) how long does it take for the greater assimilation with OSE to occur and stabilize? A long-term (224 day) Se dietary supplementation trial was conducted with serial sampling performed (days 28, 56, 112, and 224) to determine the length of time required to achieve Se supplement (OSE, Mix, and ISe)-dependent changes in Se assimilation in blood fractions and liver tissue. Forty maturing Angus heifers were fed a corn silage-based diet for 98 days with no Se supplementation, and then a cracked corn/cottonseed hull-based diet (basal diet) without Se supplementation for 74 days. Liver biopsies were taken for Se analysis, and heifers were fed the same diet for another 14 days. Heifers were assigned (n = 10) to one of four Se treatment groups such that basal liver Se contents were stratified among groups, and then fed enough of the basal diet (0.08 mg Se per day) and a mineral-vitamin mix that provided 0.16 (control) or 3.0 mg Se per day in ISe (sodium selenite), OSe (Sel-Plex(®)), or Mix (1:1 ISe:OSe) form to support 0.5 kg/day growth for 224 days. More Se was found in whole blood, red blood cells, serum, and liver of Mix and OSe heifers than ISe heifers, and all were greater than control. Se content either increased until day 56 then was stable (liver and plasma), or was stable until day 56 (whole blood) or day 112 (red blood cells) and then increased steadily through day 224, for all supplemental Se treatments. These data indicate that a 1:1 mix (1.5 mg Se:1.5 mg Se) of supplemental ISe and OSe is equal to 3 mg/day OSe supplementation and greater than 3 mg/day ISe supplementation. The data also indicate that Se levels stabilized in liver and plasma by 56 to 112 days whereas whole blood and red blood cell concentrations were still increasing through 224 days of supplementation, regardless of the form of supplemental Se.

Dietary supplementation of selenium in inorganic and organic forms differentially and commonly alters blood and liver selenium concentrations and liver gene expression profiles of growing beef heifers.

In geographic regions where selenium (Se) soil concentrations are naturally low, the addition of Se to animal feed is necessary. Even though it is known that Se in grass and forage crops is primarily present in organic forms (especially as L-selenomethionine, L-selenocystine, and L-selenocystathionine), the feeding of Se in the naturally occurring organic selenium (OSe) compounds produces higher blood and tissue Se levels than the inorganic Se (ISe) salts, and that animal metabolism of OSe and ISe is fundamentally different. Se is commonly added in inorganic form as sodium selenite to cattle feeds because it is a less expensive source of supplemental Se then are OSe forms. A trial was conducted with growing cattle to determine if the addition of OSe versus ISe forms of Se in beef cattle feed produces differences in hepatic gene expression, thereby gaining insight into the metabolic consequence of feeding OSe versus ISe. Thirty maturing Angus heifers (261 ± 6 days) were fed a corn silage-based diet with no Se supplementation for 75 days. Heifers (body weight = 393 ± 9 kg) then were randomly assigned (n = 10) and fed Se supplements that contained none (control) or 3 mg Se/day in ISe (sodium selenite) or OSe (Sel-Plex®) form and enough of a common cracked corn/cottonseed hull-based diet (0.48 mg Se/day) to support 0.5 kg/day growth for 105 or 106 days. More Se was found in jugular whole blood and red blood cells and biopsied liver tissue of ISe and OSe treatment animals than control animals, and OSe animals contained more Se in these tissues than did ISe. Microarray and bioinformatic analyses of liver tissue gene expression revealed that the content of at least 80 mRNA were affected by ISe or OSe treatments, including mRNA associated with nutrient metabolism; cellular growth, proliferation, and immune response; cell communication or signaling; and tissue/organ development and function. Overall, three Se supplement-dependent gene groups were identified: ISe-dependent, OSe-dependent, and Se form-independent. More specifically, both forms of supplementation appeared to upregulate mitochondrial gene expression capacity, whereas gene expression of a protein involved in antiviral capacity was downregulated in ISe-supplemented animals, and OSe-supplemented animals had reduced levels of mRNA encoding proteins known to be upregulated during oxidative stress and cancerous states.