Effects of body condition score (BCS) on steroid- and eicosanoid-metabolizing enzyme activity in various mare tissues during winter anoestrus.

The objective of this study was to determine the activity of steroid- and eicosanoid-metabolizing enzymes in horses with varying BCSs. The BCSs of twenty non-pregnant, anoestrous mares were determined prior to euthanasia, and tissue samples were collected from the liver, kidney, adrenal gland, ovary and endometrium. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A) and uridine 5'-diphospho-glucuronosyltransferase (UGT) activities were determined using luminogenic substrates. The MIXED procedure of SAS was used to test the effect of BCS on enzyme activity and differences between tissues. Activity of CYP1A in adrenals was increased (p ≤ .05) in BCS 5 versus BCSs 4 and 6. Activity of CYP1A in the liver was increased (p = .05) in BCS 4 versus BCSs 5 and 6. Activity of CYP1A was 100-fold greater (p < .0001) in the liver than in the adrenal, ovary and kidney. Activity of CYP2C was 100-fold greater (p < .0001) in the liver than in the adrenal, ovary and endometrium. Activity of CYP3A was only detectable in the liver. Activity of UGT in the kidney was decreased (p = .02) in BCS 4 versus BCSs 5 and 6. Activity of UGT was threefold greater (p < .0001) in the liver than in the kidney, whereas activity of UGT was ninefold greater (p < .0001) in the kidney than in the ovary and endometrium. In general, BCS did not alter the activity of steroid- and eicosanoid-metabolizing enzymes in horses. However, tissue differences in these enzymes indicated abundant hepatic metabolism in horses, which is similar to other livestock species.

Effects of maternal nutrient restriction during gestation on bovine serum microRNA abundance.

The objective was to determine the effects of maternal nutrient restriction during gestation on serum microRNA (miRNA) abundance in cattle. Primiparous Angus-cross cows (n=22) were fed either control (CON; to gain 1 Kg/week) or nutrient restricted (NR; 0.55% NEm) diets based on National Research Council requirements. On day 30 of gestation, cows were blocked by body condition and randomly assigned to one of three diets: CON (n=8) days 30-190; NR (n=7) days 30-110 followed by CON days 110-190 (NR/C); or CON (n=7) days 30-110 followed by NR days 110-190 (C/NR). At 190 days of gestation, maternal serum was collected for RNA isolation and analyzed using a miRNA microarray of known Bos taurus sequences. Data were normalized using LOWESS and analyzed via ANOVA. At 190 days of gestation, 16 miRNAs exhibited differential abundance (P<0.05) between treatments. Cows that underwent NR, irrespective of when the insult occurred, had downregulated bta-miR-126-3p compared to CON cows. Bta-miR-16b was downregulated and three miRNAs upregulated in NR/C compared to C/NR and CON cows. Additionally, seven miRNAs were downregulated and four miRNAs upregulated in C/NR compared to NR/C and CON cows. Comparison of NR/C and C/NR cows revealed three differentially abundant (P<0.04) miRNAs (bta-miR-2487_L-2R-3_1ss15CT, bta-miR-215, and bta-miR-760-5p). Top KEGG pathway enrichment of target genes included: pathways in cancer, PI3K-Akt signaling, focal adhesion, Ras signaling, proteoglycans in cancer, and MAPK signaling. In summary, maternal nutrient restriction altered serum miRNA abundance profiles irrespective of the time at which the nutritional insult was induced.

Effects of post-weaning supplementation of immunomodulatory feed ingredient on body weight and cortisol concentrations in program-fed beef heifers.

The objective of this study was to determine the effects of an immunomodulatory feed ingredient during post-weaning on growth and cortisol concentrations of beef heifers. Commercial Angus heifers (n = 72) from 2 AI sires were blocked (n = 9) by BW and randomly assigned to one of 2 pens per block. Each pen (4 heifers/pen) per block was randomly assigned to either one of 2 treatments, Celmanax (CEL) or corn germ (CON). The heifers were fed a commercial total mixed ration (TMR) twice daily from d 0 to 60 to gain 0.75 kg/d. The feed was top-dressed once daily with either 72g of Celmanax (CEL) or corn germ (CON) per pen. Body weight (BW) was collected on d -1, 0, 15, 30, 45, 60, and 61. Blood samples were collected on d 0, 15, 30, 45, and 60. Two heifers per pen (n = 32) were randomly selected for a transportation challenge to evaluate stress response on d 62 or 63 of the study. Sixteen heifers (n = 8 CEL; n = 8 CON) were randomly selected for a corticotropin-releasing hormone/arginine vasopressin (CRH/AVP) challenge and an intravenous glucose tolerance test (IVGTT) on d 64 and 67 of the study. The pen was the experimental unit, and data were analyzed by ANOVA or repeated measures analysis as appropriate. Feed efficiency and BW gain were increased (P = 0.04) in CEL heifers compared to CON heifers. Serum cortisol concentrations were decreased (P < 0.01) in CEL heifers compared to CON heifers on d 30 to 60 post-weaning. Serum cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers throughout the transportation challenge. Serum cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers during the CRH/AVP challenge from 60 to 150 min post-infusion. Treatment did not influence (P = 0.29) plasma insulin or glucose (P = 0.63) concentrations during the IVGTT. In summary, supplementation of Celmanax post-weaning increased BW gain and reduced cortisol concentrations in challenged beef heifers.

The effects of early- or mid-gestation nutrient restriction on bovine fetal pancreatic development.

Research on the effects of nutrient restriction in beef cows on fetal pancreatic development is limited. To address this, multiparous Angus-cross cows (n = 22) were fed either control (CON; to gain 1 kg/wk) or nutrient-restricted (NR; 0.55% NEm) diets based on NRC requirements. On d 30 of gestation, cows were blocked by body condition and randomly assigned to one of three nutritional regimes: CON fed from d 30 to 190 (n = 8), or NR/C (n = 7) or C/NR (n = 7) fed either the CON or NR diet from d 30 to 110 followed by CON or NR from d 110 to 190 of gestation. Cows were harvested on d 190 of gestation, and blood samples, fetal weights, and fetal tissue weights and samples were collected. Pancreas samples were embedded in paraffin and sectioned for standard immunohistochemistry procedures to quantify insulin-positive ß cells and number of apoptotic ß cells using TUNEL staining. Data were analyzed via ANOVA using the general linear model procedure of SAS. At harvest, empty carcass weights were decreased (P = 0.036) in fetuses of C/NR and NR/C fed dams compared to fetuses of CON fed dams. Pancreas weight was decreased (P = 0.028) in fetuses of C/NR fed dams compared to CON fetuses; however, fetuses of NR/C fed dams were not different (P > 0.05) from fetuses of CON fed dams. Maternal and fetal serum insulin concentrations were not different (P > 0.05) in NR/C fed compared to CON fed; however, concentrations of insulin were decreased (P = 0.036 and P = 0.40, respectively) in C/NR fed compared to CON fed. Beta cell number was decreased (P = 0.009) in fetuses of NR/C and C/NR fed dams compared to fetuses of CON fed dams. Percentage of apoptotic cells was increased (P < 0.0001) in fetuses of NR/C and C/NR fetuses than fetuses of CON fed dams. This evidence suggests that nutrient restriction either during early- or mid-gestation can negatively impact fetal pancreatic development. However, mid-gestational nutritional insult is potentially recovered by reacclimation to a diet that meets requirements of the dam, thus reducing negative outcomes in fetal offspring.

Endometrial blood perfusion as assessed using a novel laser Doppler technique in Angus cows.

Previous studies have characterized ovarian steroid synthesis which directly affects uterine environment and blood flow. Clearance of steroids occurs primarily in hepatic tissues, however, it was discovered that there is an abundant activity of the phase II steroid metabolizing enzyme UDP-glucuronosyltransferase (UGT) in uterine biopsies. No minimally invasive techniques for collecting endometrial perfusion, which is affected by steroids and indicative of reproductive health, have been developed for livestock. The objective of the present study was to characterize UGT activity and endometrial blood perfusion during a normal estrous cycle of cattle. It was hypothesized that there would be increased steroid metabolism during the luteal phase of the estrous cycle and in the uterine horn ipsilateral to the corpus luteum (CL). During the first synchronized estrous cycle, progesterone and UGT activity increased on Day 6 compared with 0 and 3, with the first day of estrus being considered Day 0 of the study. Endometrial perfusion was greater ipsilateral to the CL compared with contralateral on Day 12, and was less ipsilateral to the CL compared with contralateral on Day 18. Similar to perfusion results, nitric oxide metabolites (nitrites) were greatest in the endometrium ipsilateral as compared with contralateral to the CL. Moreover, there was a positive correlation (r = 0.28; P = .04) between endometrial perfusion and nitrite concentration. It is concluded that activity of UGT within the endometrium is affected by the contralateral or ipsilateral location of the CL, and collection of endometrial perfusion data using a laser Doppler probe could be a viable measurement technique as indicated by associated nitrite concentrations in the present study.