Effect of Post-Weaning Concentrate Feeding Prior to Forage Finishing on Intramuscular Fat Deposition.

The objectives of this study were to examine the effects of feeding high-concentrate diets post-weaning (PW) prior to forage finishing on (1) changes in ultrasound intramuscular fat deposition and lipogenic/lipolytic gene expression during the post-weaning phase and (2) carcass characteristics and fatty acid composition after forage finishing to 487 kg. Steers were randomly assigned to one of four treatments (PW0, PW40, PW80, and PW120) at weaning to examine the time of high-concentrate feeding prior to forage finishing. The ultrasound intramuscular fat content was greater (p < 0.05) for PW120 compared to those for PW0, PW40, or PW80 at the end of the post-weaning phase. Feeding high concentrates (PW120) up-regulated (p < 0.01) the mRNA expression of fatty acid transporters and lipogenic genes and down-regulated lipolytic genes in the LM compared to PW0. Carcasses from PW120 were graded 83% Choice (p = 0.025), whereas carcasses from other post-weaning treatments (PW0, 40, or 80) were graded 25, 36, and 54% Choice, respectively, at the final harvest. The total fatty acid content of the muscle at slaughter was greater (p = 0.0004) for PW120 than PW0, PW40, and PW80. Feeding high-concentrate diets to steers post-weaning for 120 day enhanced early intramuscular fat deposition without causing major changes to the fatty acid composition of the longissimus muscle after forage finishing.

Effects of Dam and Sire Breeds on Lamb Carcass Quality and Composition in Pasture-Based Systems.

This study explored the impacts of sire and dam breed on carcass quality and composition in a pasture-based system and the use of DXA to rapidly rank carcasses for leanness. Southdown (SD) and Suffolk (SF) ewes were mated to Texel (TX) or SD rams to produce seventy-nine lambs. Lambs were raised on pasture-based systems with limited grain supplementation. Lamb birth weight was greater (p < 0.01) for TX, regardless of dam breed. Lambing rate was lower (p < 0.01) for SD than SF ewes. Circulating myostatin concentrations were greater (p < 0.05) on d 42 than d 75 or d 110 but did not differ by sire breed. Texel-sired lambs had greater (p < 0.01) carcass weight, ribeye area and quality grade compared to SD-sired. Total and primal fat mass as predicted from DXA was higher (p < 0.05) in carcasses from SD than TX sires. Muscles from TX lambs had greater (p < 0.05) polyunsaturated fatty acid (PUFA) composition than SD-sired. Shear force values were influenced (p < 0.01) by dam breed, muscle cut and postmortem age but not by sire breed. The use of TX sires in pasture-based systems improved carcass leanness and muscle PUFA concentrations without altering tenderness.

Training and Adaptation of Beef Calves to Precision Supplementation Technology for Individual Supplementation in Grazing Systems.

Supplementation of beef cattle can be used to meet both nutrient requirements and production goals; however, supplementation costs influence farm profitability. Common supplementation delivery strategies are generally designed to provide nutrients to the mean of the group instead of an individual. Precision individual supplementation technologies, such as the Super SmartFeed (SSF, C-Lock Inc., Rapid City, SD, USA), are available but are generally cost prohibitive to producers. These systems require adaptation or training periods for cattle to utilize this technology. The objective of this research was to assess the training and adoption rates of three different groups of cattle (suckling calves, weaned steers, replacement heifers) to the SSF. Successful adaptation was determined if an individual's supplement intake was above the group average of total allotted feed consumed throughout the training period. Suckling calves (n = 31) underwent a 12 d training period on pasture; 45% of suckling calves adapted to the SSF and average daily intake differed (p < 0.0001) by day of training. Weaned steers (n = 79) were trained in drylot for 13 d. Of the weaned steers, 62% were trained to the SSF, and average daily intake differed (p < 0.0001) by day of training. Replacement heifers (n = 63) grazed tall fescue pastures and had access to SSF for 22 d of training. The success rate of replacement heifers was 73%. For replacement heifers, the daily intake did not differ (p < 0.0001) by day of training. Results indicate production stage may influence cattle adaptation to precision technologies.

Identification of microRNA Transcriptome Involved in Bovine Intramuscular Fat Deposition.

Background: Intramuscular fat deposition in beef is a major determinant of carcass quality and value in the USA. The objective of this study was to examine changes in microRNA (miRNA) transcriptome that are involved with intramuscular fat deposition with time-on-concentrates (TOC). Yearling steers were individually fed a high concentrate diet and changes in intramuscular fat deposition were monitored by real-time ultrasound at 28 to 33 d intervals. Longissimus muscle biopsies collected on d 0, 92 and 124 TOC to examine changes in miRNA transcriptome that are involved in intramuscular fat deposition. Results: Steer body weight increased (P < 0.0001) at each weigh day during TOC. Fat thickness increased (P < 0.005) from d 28 to 124. Ribeye area was larger (P < 0.001) on d 124 than d 61, which was larger than d 0 and 28. Ultrasound intramuscular fat content was greater (P < 0.001) on d 92 and 124 compared to d 0, 28 or 61. Sequencing of the muscle biopsy samples identified one miRNA, bta-miR-122, that was up-regulated (P < 0.005) at d 92 and 124 compared to d 0. At d 92 TOC, mRNA expression levels of fatty acid binding protein 4 (FABP4) and elongase 6 (ELOVL6) were up-regulated (P < 0.01) compared to d 0; whereas at d 124, lipogenic genes involved in de novo fatty acid synthesis, fatty acid transport, elongation and desaturation were highly up-regulated compared to d0. Conclusions: Small RNA sequencing identified bta-miR-122 as a potential miRNA of interest that may be involved in intramuscular fat deposition with increasing TOC. Increased intramuscular fat content, as measured by real-time ultrasound, combined with differential gene expression suggests that preadipocyte differentiation may be stimulated first, which is followed by a global up-regulation of lipogenic genes involved in de novo fatty acid synthesis that provide fatty acids for subsequent hypertrophy.

Effects of Tall Fescue Endophyte Type and Dopamine Receptor D2 Genotype on Cow-Calf Performance during Late Gestation and Early Lactation.

Grazing endophyte-infected, toxic tall fescue reduces cow/calf production; therefore, this study examines alternate strategies such as use of novel endophyte fescue varieties during late gestation and early lactation or genetic selection of resistant cows. Pregnant cows (n = 75) were randomly assigned to fescue endophyte type: 1) endophyte-infected ergot alkaloid producing tall fescue (E+) or 2) novel endophyte-infected, non-toxic tall fescue (NOV) within maternal (A|A, n = 38 and G|G, n = 37) DRD2 genotype to examine changes in cow/calf performance and milk production during late gestation and early lactation. Grazing E+ fescue pastures during late gestation reduced cow body weight gain but did not alter calf birth weight compared to NOV. Milk production and calf ADG during the first 30 day of lactation were lower for E+ than NOV. The calving rate was reduced, but not calving interval for E+ cows. The adjusted 205-day weight of calves was lower in those grazing E+ with their dams compared to NOV. There were no interactions between DRD2 genotype and fescue endophyte type indicating that genotype was not associated with response to E+ fescue in this study. Overall, grazing NOV tall fescue pastures rather than E+ during critical stages of production improved cow gain during late gestation, calving rate, early milk production and calf growth.

Feeding Tall Fescue Seed Reduces Ewe Milk Production, Lamb Birth Weight and Pre-Weaning Growth Rate.

Endophyte-infected tall fescue (E+) produces ergovaline and ergovalinine, which are mycotoxins that act as dopamine agonists to suppress prolactin and induce vasoconstriction. The experiment was designed as a 3 × 2 × 2 factorial with DRD2 genotype (AA, AG, GG), fescue seed (endophyte-free, E- or endophyte-infected, E+), stage of gestation (MID, d (day) 35-85; LATE, d 86-parturition) and all interactions in the model. Pregnant Suffolk ewes (n = 60) were stratified by genotype and fed E+ or E- seed in a total mixed ration according to treatment assignment. Serum prolactin concentrations were lower (p < 0.05) in ewes fed E+ seed but did not differ by maternal DRD2 genotype or two-way interaction. Lamb birth weight was lower (p < 0.05) in ewes fed E+ seed in last trimester. Pre-weaning growth rate, milk production and total weaning weight was reduced (p < 0.05) in ewes fed E+ fescue seed during MID and LATE gestation. Ingestion of ergovaline/ergovalinine in last trimester reduces lamb birth weight; however, lamb growth rate, milk production and total weaning weight are reduced in all ewes fed E+ during mid and last trimester.

Evaluation of oral citrulline administration as a mitigation strategy for fescue toxicosis in sheep.

Gestating ewes consuming ergot alkaloids, from endophyte-infected (E+) tall fescue seed, suffer from intrauterine growth restriction and produce smaller lambs. Arginine (Arg) supplementation has been shown to increase birth weight and oral citrulline (Cit) administration is reported to increase arginine concentrations. Two experiments were conducted to: 1) evaluate if oral supplementation with Cit or water, to ewes consuming E+ fescue seed, increases lamb birth weight and 2) determine the effectiveness of Cit and citrulline:malate as an oral drench and elevating circulating levels of Cit to determine levels and dose frequency. In experiment 1, gestating Suffolk ewes (n = 10) were assigned to one of two treatments [oral drench of citrulline-malate 2:1 (CITM; 81 mg/kg/d of citrulline) or water (TOX)] to start on d 86 of gestation and continued until parturition. Ewes on CITM treatment had decreased (P < 0.05) plasma Arg and Cit concentrations during gestation. At birth, lambs from CITM ewes had reduced (P < 0.05) crude fat and total fat but did not differ (P > 0.05) in birth weight from lambs born to TOX ewes. In experiment 2, nonpregnant Suffolk ewes (n = 3) were assigned to either oral citrulline (CIT; 81 mg/kg/d), citrulline-malate 2:1 (CITM; 81 mg/kg/d of citrulline), or water (CON) drench in a Latin Square design for a treatment period of 4 d with a washout period of 3 d. On d 4, blood samples were collected at 0, 0.5, 1, 2, 3, 4, 6, 8, 10, 12, and 18 h post drench. Oral drenching of CIT and CITM increased (P < 0.0001) Cit concentrations within 2 h and levels remained elevated for 6 h. Apparent half-life of elimination for CIT and CITM were 8.484 and 10.392 h, respectively. Our results show that lamb birth weight was not altered with a single oral drench of citrulline-malate; however, lamb body composition was altered. The level and frequency of citrulline dosing may need to be greater in order to observe consistent elevation of Cit/Arg concentrations to determine its effectiveness in mitigating fescue toxicosis.

Feeding Tall Fescue Seed during Mid and Late Gestation Influences Subsequent Postnatal Growth, Puberty, and Carcass Quality of Offspring.

Weaned lambs (n = 82), born to ewes fed endophyte-free (E-) or endophyte-infected (E+; 1.77 mg hd-1 d-1 ergovaline + ergovalinine) tall fescue seed from d 35 to 85 of gestation (MID) and/or d 86 of gestation to parturition (LATE), were used to examine how ergot alkaloid exposure during fetal development altered subsequent puberty attainment or carcass quality. Lambs were weaned at 75 d of age and separated by sex to assess puberty in ewe lambs (n = 39) and to evaluate growth, carcass and meat quality in wethers (n = 43). Data were analyzed with maternal fescue treatment, stage of gestation, and two-way interaction in the model. Age at puberty tended (P = 0.06) to be longer for ewe lambs born to dams fed E+ fescue during LATE gestation versus those fed E-. Post-weaning average daily gain tended to be higher (P = 0.07) for wether lambs born to dams fed E+ fescue seed during MID gestation compared to E-. Exposure to ergot alkaloids during fetal growth altered (P < 0.10) longissimus muscle weight and color, lipid deposition, fatty acid composition, and shear force values of semimembranosus muscle in wether lambs. These results indicate that exposure to ergot alkaloids in utero does alter subsequent post-weaning puberty attainment and body composition in offspring.

Flaxseed Oil or n-7 Fatty Acid-Enhanced Fish Oil Supplementation Alters Fatty Acid Composition, Plasma Insulin and Serum Ceramide Concentrations, and Gene Expression in Lambs.

The objective of this study was to examine the effects of flaxseed (FLAX) oil or 16-carbon n-7 fatty acid -enhanced fish oil (Provinal; POA) supplementation on serum, liver and skeletal muscle fatty acid concentrations, serum ceramide and plasma insulin concentrations, and gene expression. Lambs [n = 18; 42 ± 5.6 kg body weight (BW); 7 months] were individually fed one of the three treatments: (1) control (CON), no oil supplement, (2) FLAX; at 0.1% of BW, or (3) POA at 0.1% of BW for 60 days. Daily feed intake and weight gain were decreased by 21% and 34%, respectively, for POA than FLAX. Liver and skeletal muscle concentrations of palmitoleic acid were greater by 396% and 87%, respectively, for POA than FLAX; whereas, liver and skeletal muscle α-linolenic acid concentrations were greater by 199% and 118%, respectively, for FLAX. Supplementation with POA also had greater serum and tissue concentrations of eicosapentaenoic and docosahexaenoic acids. Serum glucose and plasma insulin concentrations were elevated with FLAX supplementation at the end of the study. Supplementation with POA altered serum ceramide concentrations compared to CON or FLAX. Oil supplementation, both FLAX and POA, downregulated expression of unesterified fatty acid receptors (FFAR) 1 and FFAR4 in the liver; however, oil supplementation upregulated expression of FFAR1 in muscle. Interleukin-6 (IL6) and tumor necrosis factor-α (TNFA) expression were downregulated with oil supplementation in the liver; however, FLAX upregulated TNFA in muscle. These results show that oil supplementation can enhance uptake and deposition of unique fatty acids that alter ceramide concentrations and gene expression in tissues.

Ergot alkaloid exposure during gestation alters: 3. Fetal growth, muscle fiber development, and miRNA transcriptome1.

The objective of this study was to assess how exposure to ergot alkaloids during 2 stages of gestation alters fetal growth, muscle fiber formation, and miRNA expression. Pregnant ewes (n = 36; BW = 83.26 ± 8.14 kg; 4/group; 9 groups) were used in a 2 × 2 factorial arrangement with 2 tall fescue seed treatments [endophyte-infected (E+) vs. endophyte-free (E-)] fed during 2 stages of gestation (MID, days 35 to 85 vs. LATE, days 86 to 133), which created 4 possible treatments (E-/E-, E+/E-, E-/E+, or E+/E+). Ewes were individually fed a total mixed ration containing E+ or E- fescue seed according to treatment assignment. Terminal surgeries were conducted on day 133 of gestation for the collection of fetal measurements and muscle samples. Data were analyzed as a 2 × 2 factorial with fescue treatment, stage of gestation, and 2-way interaction as fixed effects. Fetuses exposed to E+ seed during LATE gestation had reduced (P = 0.0020) fetal BW by 10% compared with E- fetuses; however, fetal body weight did not differ (P = 0.41) with E+ exposure during MID gestation. Fetuses from ewes fed E+ seed during MID and LATE gestation tended to have smaller (P = 0.058) kidney weights compared with E- fetuses. Liver weight was larger (P = 0.0069) in fetuses fed E- during LATE gestation compared with E+. Fetal brain weight did not differ by fescue treatment fed during MID (P = 0.36) or LATE (P = 0.40) gestation. The percentage of brain to empty body weight (EBW) was greater (P = 0.0048) in fetuses from ewes fed E+ fescue seed during LATE gestation, which is indicative of intrauterine growth restriction (IUGR). Primary muscle fiber number was lower (P = 0.0005) in semitendinosus (STN) of fetuses exposed to E+ during MID and/or LATE gestation compared with E-/E-. miRNA sequencing showed differential expression (P < 0.010) of 6 novel miRNAs including bta-miR-652_R+1, mdo-miR-22-3p, bta-miR-1277_R-1, ppy-miR-133a_L+1_1ss5TG, hsa-miR-129-1-3p, and ssc-miR-615 in fetal STN muscle. These miRNA are associated with glucose transport, insulin signaling, intracellular ATP, hypertension, or adipogenesis. This work supports the hypothesis that E+ tall fescue seed fed during late gestation reduces fetal weight and causes asymmetrical growth, which is indicative of IUGR. Changes in primary fiber number and miRNA of STN indicate that exposure to E+ fescue fed during MID and LATE gestation alters fetal muscle development that may affect postnatal muscle growth and meat quality.

Ergot alkaloid exposure during gestation alters. I. Maternal characteristics and placental development of pregnant ewes1.

Tall fescue [Lolium arundinaceum (Scheyreb.) Darbysh] is the primary cool season forage grass in the Southeastern United States. Most tall fescue contains an endophytic fungus (Epichloë coenophiala) that produces ergot alkaloids and upon ingestion induces fescue toxicosis. The objective of this study was to assess how exposure to endophyte-infected (E+; 1.77 mg hd-1 d-1 ergovaline and ergovalinine) or endophyte-free (E-; 0 mg hd-1 d-1 ergovaline and ergovalinine) tall fescue seed fed during 2 stages of gestation (MID, days 35-85/LATE, days 86-133) alters placental development. Thirty-six, fescue naïve Suffolk ewes were randomly assigned to 1 of 4 fescue treatments: E-/E-, E-/E+, E+/E-, or E+/E+. Ewes were individually fed the same amount of E+ or E- seed mixed into total mixed ration during MID and LATE gestation. Terminal surgeries were conducted on day 133 of gestation. Ewes fed E+ fescue seed had elevated (P < 0.001) ergot alkaloid excretion and reduced (P < 0.001) prolactin levels during the periods when fed E+ seed. Ewes switched on day 86 from E- to E+ seed had a 4% reduction (P = 0.005) in DMI during LATE gestation, which translated to a 2% reduction (P = 0.07) in DMI overall. Average daily gain was also reduced (P = 0.049) by 64% for E-/E+ ewes during LATE gestation and tended to be reduced (P = 0.06) by 33% overall. Ewes fed E+ seed during LATE gestation exhibited a 14% and 23% reduction in uterine (P = 0.03) and placentome (P = 0.004) weights, respectively. Caruncle weights were also reduced by 28% (P = 0.003) for E-/E+ ewes compared with E-/E- and E+/E-. Ewes fed E+ seed during both MID and LATE gestation exhibited a 32% reduction in cotyledon (P = 0.01) weights, whereas ewes fed E+ seed only during MID gestation (E+/E-) had improved (P = 0.01) cotyledon weights. The percentage of type A placentomes tended to be greater (P = 0.08) for E+/E+ ewes compared with other treatments. Other placentome types (B, C, or D) did not differ (P > 0.05). Total fetal weight per ewe was reduced (P = 0.01) for ewes fed E+ seed during LATE gestation compared with E-; however, feeding E+ seed during MID gestation did not alter (P = 0.70) total fetal weight per ewe. These results suggest that exposure to ergot alkaloids during LATE (days 86-133) gestation has the greatest impact on placental development by reducing uterine and placentome weights. This, in turn, reduced total fetal weight per ewe by 15% in ewes fed E+ seed during LATE gestation (E-/E+ and E+/E+).

Ergot alkaloid exposure during gestation alters: II. Uterine and umbilical artery vasoactivity1.

Previous research has shown that livestock exposed to ergot alkaloids results in decreased vasoactivity of gastrointestinal and peripheral vasculature. Little is known regarding the effect ergot alkaloid exposure during gestation may have on vasculature supporting the fetus. The objective of this study was to evaluate contractile responses of uterine and umbilical arteries collected from ewes consuming ergot alkaloids during gestation. On day 35 of gestation, 36 Suffolk ewes (78.24 ± 9.5 kg) were assigned to endophyte-infected (E+) or endophyte-free (E-) tall fescue seed treatments that were fed either throughout or switched on day 86 of gestation, creating four seed treatments E+E+, E+E-, E-E+, and E-E-. Ewes were fed E+ tall fescue seed to provide 1.77 mg of total ergovaline ⋅ hd-1 ⋅ d-1 with E- ewes receiving the same quantity of E- seed. Gestation was terminated on day 133, and sections of uterine artery and umbilical cord were surgically collected. Only collections from 28 ewes (n = 7/treatment) were of sufficient viability to proceed with the contractility experiments. Arteries were cleaned, sliced into 2-mm cross sections, and suspended in multi-myograph chambers containing 5 mL of continuously oxygenated Krebs-Henseleit buffer. Vessels were exposed to increasing concentrations (5 × 10-8 to 1 × 10-4 M) of norepinephrine, serotonin, ergotamine, and ergovaline (5 × 10-9 to 1 × 10-5M; extract of tall fescue seed) in 15-min intervals. Increasing concentrations of norepinephrine generated a contractile response by the uterine artery (P < 0.05), but no response in the umbilical artery. Increasing concentrations of serotonin resulted in negligible responses in uterine preparations, whereas umbilical artery preparations were responsive (P < 0.05) to serotonin. Ewes receiving E+E+ and E-E+ treatments had decreased vasoactivity in umbilical arteries to serotonin with a dextral shift in concentrations where the response curve initiated (P < 0.05). Interestingly, uterine arteries were not responsive to exposure to ergotamine or ergovaline, whereas umbilical arteries were responsive (P < 0.05). Umbilical arteries collected from ewes receiving E-E- and E+E- were more vasoactive to ergot alkaloids (P < 0.05) than other treatments. These findings indicate that maternal blood supply to the placenta appears protected from negative effects of ergot alkaloids; however, umbilical vasculature is not, and this could adversely influence fetal growth.

Supplementation of glycerol or fructose via drinking water to enhance marbling deposition and meat quality of finishing cattle.

Thirty-six Angus-cross steers (667 ± 34.4 kg initial BW, 24.5 mo) were used to assess the impact of short-term glycerin or high-fructose corn syrup administration via drinking water on meat quality and marbling deposition. Steers blocked by BW (3 blocks) were assigned randomly to 1 of 3 drinking water treatments: 1) control (CON), 2) 4.3% crude glycerin (GLYC), or 3) 4.3% high-fructose corn syrup (HFCS) for the final 25 d before slaughter. Average daily gain was lower ( = 0.01) and final live weight was lower ( < 0.01) with HFCS administration compared with CON. Dry matter intake and water intake did not differ among treatments. Fat thickness, muscle depth, and intramuscular fat measured by ultrasound did not differ among treatments. Crude glycerin or HFCS via water supplementation did not alter HCW, dressing percentage, rib eye area, fat thickness, KPH, skeletal maturity, or marbling score. Longissimus muscle and subcutaneous fat color (L*, a*, and b*) were not affected by drinking water treatment. Total lipid content, total fatty acid content, and fatty acid composition of the LM did not differ among drinking water treatments. Supplementation of drinking water with GLYC or HFCS did not alter Warner-Bratzler shear force values or water-holding capacity (drip loss, cook shrink). Intramuscular mean adipocyte diameter was greater ( = 0.02) for steers offered HFCS compared with steers offered GLYC, with CON steers being intermediate. These differences in mean adipocyte size were related to changes in the adipocyte size distribution. There were greater proportions of small (20 to 30 µm) adipocytes in GLYC compared with HFCS and CON. In contrast, HFCS and CON had greater proportions of medium (40 to 50 µm) adipocytes than GLYC. The relative mRNA expression of lipogenic genes (acetyl Co-A carboxylase [ACC], fatty acid binding protein 4 [FABP4], fatty acid synthase [FASN], glycerol-3-phosphate acyltransferase [GPAT], retinol-binding protein 4 [RBP4], and stearoyl-CoA desaturase [SCD]), adipocyte differentiation genes (delta-like 1 homolog [DLK1]), and transcription factors (CCAAT/enhancer-binding protein α [C/EBPα], and PPARγ) was similar for GLYC and HFCS compared with CON. Longissimus glycogen and lactate concentrations and glycolytic potential were not affected by drinking water treatments. Overall, HFCS or GLYC supplementation via drinking water did not alter carcass or meat quality variables but did alter the size and distribution of intramuscular adipocytes. These results indicate that a longer supplementation time or a higher substrate level may be needed to obtain differences in meat quality.

The bull sperm microRNAome and the effect of fescue toxicosis on sperm microRNA expression.

Tall fescue [Schedonorus phoenix (Scop.) Holub] accounts for nearly 16 million hectares of pasture in the Southeastern and Mid-Atlantic U.S. due to its heat, drought, and pest resistance, conferred to the plant by its symbiotic relationship with the endophyte Neotyphodium coenophialum. The endophyte produces ergot alkaloids that have negative effects on the growth and reproduction of animals, resulting in the syndrome known as fescue toxicosis. The objectives of our study were to identify microRNA (miRNA) present in bovine sperm and to evaluate the effects of fescue toxicosis on sperm miRNA expression. Angus bulls were assigned to treatments of either toxic or non-toxic fescue seed diets. Semen was collected and subjected to RNA isolation. Three samples from each treatment group were chosen and pooled for deep sequencing. To compare miRNA expression between treatment groups, a microarray was designed and conducted. For each of the top ten expressed miRNA, target prediction analysis was conducted using TargetScan. Gene ontology enrichment was assessed using the Database for Annotation, Visualization and Integrated Discovery. Sequencing results elucidated the presence of 1,582 unique small RNA present in sperm. Of those sequences, 382 were known Bos taurus miRNA, 22 were known but novel to Bos taurus, and 816 were predicted candidate miRNA that did not map to any currently reported miRNA. Of the sequences chosen for microarray, twenty-two showed significant differential expression between treatment groups. Gene pathways of interest included: regulation of transcription, embryonic development (including blastocyst formation), Wnt and Hedgehog signaling, oocyte meiosis, and kinase and phosphatase activity. MicroRNA present in mature sperm appears to not only be left over from spermatogenic processes, but may actually serve important regulatory roles in fertilization and early developmental processes. Further, our results indicate the possibility that environmental changes may impact the expression of specific miRNA.

Palmitoleic acid reduces intramuscular lipid and restores insulin sensitivity in obese sheep.

Obese sheep were used to assess the effects of palmitoleic (C16:1 cis-9) acid infusion on lipogenesis and circulating insulin levels. Infusion of 10 mg/kg body weight (BW)/day C16:1 intravenously in obese sheep reduced (P<0.01) weight gain by 77%. Serum palmitoleic levels increased (P<0.05) in a linear manner with increasing levels of C16:1 infusion. Cis-11 vaccenic (C18:1 cis-11) acid, a known elongation product of palmitoleic acid, was also elevated (P<0.05) in serum after 14 days and 21 days of infusion. Plasma insulin levels were lower (P<0.05) (10 mg/kg BW/day C16:1) than controls (0 mg/kg BW/day C16:1) at 14 days and 28 days of infusion. Infusion of C16:1 resulted in linear increases in tissue concentrations of palmitoleic, cis-11 vaccenic, eicosapentaenoic, and docosapentaenoic acids in a dose-dependent manner. Total lipid content of the semitendinosus (ST) muscle and mesenteric adipose tissue was reduced (P<0.01) in both 5 mg/kg and 10 mg/kg BW C16:1 dose levels. Total lipid content and mean adipocyte size in the longissimus muscle was reduced (P<0.05) in the 10 mg/kg BW C16:1 dose level only, whereas total lipid content and adipocyte size of the subcutaneous adipose tissue was not altered. Total lipid content of the liver was also unchanged with C16:1 infusion. Palmitoleic acid infusion upregulated (P<0.05) acetyl-CoA carboxylase (ACC), fatty acid elongase-6 (ELOVL6), and Protein kinase, AMP-activated, alpha 1 catalytic subunit, transcript variant 1 (AMPK) mRNA expressions in liver, subcutaneous adipose, and ST muscle compared to the controls. However, mRNA expression of glucose transporter type 4 (GLUT4) and carnitine palmitoyltransferase 1b (CPT1B) differed between tissues. In the subcutaneous adipose and liver, C16:1 infusion upregulated (P<0.05) GLUT4 and CPT1B, whereas these genes were downregulated (P<0.05) in ST muscle with C16:1 infusion. These results show that C16:1 infusion for 28 days reduced weight gain, intramuscular adipocyte size and total lipid content, and circulating insulin levels. These changes appear to be mediated through alterations in expression of genes regulating glucose uptake and fatty acid oxidation specifically in the muscles.

Exposure to ergot alkaloids during gestation reduces fetal growth in sheep.

Tall fescue [Lolium arundinaceum (Schreb.) Darbysh; Schedonorus phoenix (Scop.) Holub] is the primary cool season perennial grass in the eastern U.S. Most tall fescue contains an endophyte (Neotyphodium coenophialum), which produces ergot alkaloids that cause vasoconstriction and could restrict blood flow to the fetus in pregnant animals. The objective of this study was to examine fetal growth during maternal exposure to ergot alkaloids during gestation. Pregnant ewes (n = 16) were randomly assigned to one of two dietary treatments: (1) endophyte-infected (N. coenophialum) tall fescue seed (E+; 0.8 ug of ergovaline /g diet DM) and (2) endophyte-free tall fescue seed (E-; 0.0 ug of ergovaline/g diet DM). Birth weight of lambs was reduced by 37% for E+ compared to E-. Organ and muscle weights were also lighter for E+ than E-. Exposure to ergot alkaloids in utero reduces fetal growth and muscle development.

Nutritional milieu of isolated stromal vascular cells determines their proliferative, adipogenic, and lipogenic capacity in vitro.

The objective was to determine the effect of nutritional milieu of isolated stromal vascular (SV) cells on proliferative capacity of preadipocytes, and adipogenic and lipogenic capacity in adipocytes in vitro. Proliferation of the preadipocytes increased over time with 48 and 72 h being greater than 24 h; however, preadipocytes from steers supplemented with corn (LC) had lower proliferation rates compared with those without corn grain supplementation (L) at 72 h. Adipocyte cultures isolated from LC group had higher mean diameter on d 4 and 6, and higher mean volume on d 0, 4, 6, and 12 of culture. Adipocytes from steers supplemented with corn grain (LC) had lower expression of key adipogenic genes during extended days in culture. The results show that prior nutritional treatment of the donor animal used to isolate SV cultures alters their proliferative, adipogenic, and lipogenic capacity in culture. These differences may be related to lower induction/expression of AP2 gene in the adipose cultures from corn supplemented group. Corn grain supplementation to steers grazing legumes could have stimulated more active adipogenic progenitor cells to differentiate, which would leave fewer behind in the SV pool for subsequent isolation.

Fatty acid composition and interrelationships among eight retail cuts of grass-feed beef.

The aim of this research was to evaluate: 1) fatty acid (FA) profile of eight retail cuts (eye of round, ribeye, top round, striploin, tenderloin, top-sirloin, underblade, ground beef) from grass-fed steers, 2) the association between striploin (longissimus muscle) FA profile and that from the other retail cuts, and 3) the changes in FA profile associated with retail cut total FA content. All of the retail cuts, except underblade and ground beef, would be considered lean (<10 g) based on total FA content. Total saturated fatty acids were positively correlated to total FA content; whereas, negative correlations were observed with total FA content and total monounsaturated and polyunsaturated fatty acids. No correlation was observed between total FA and CLA cis-9, trans-11. Striploin trans-11 C18:1 and CLA cis-9, trans-11 proportions, and n-6:n-3 ratio were positive and highly correlated with those in the other retail cuts.

Isolation of total RNA and detection procedures for miRNA present in bovine-cultured adipocytes and adipose tissues.

Micro-ribonucleic acids (miRNA) regulate gene expression posttranscriptionally by altering translation of protein(s) encoded by specific messenger RNA. Therefore the ability to detect and quantify the expression levels of specific miRNA present within a cell or tissue is necessary to thoroughly examine cellular physiology and gene expression. Here we describe procedures that allow for the isolation and quantification of miRNA in bovine adipocytes and adipose tissue.

Omega-6 fat supplementation alters lipogenic gene expression in bovine subcutaneous adipose tissue.

In contrast to rodents, adipose tissue serves as the major site of lipogenesis and storage reservoir for excess dietary energy in cattle. Research in rodents shows that adding corn oil (57% C18:2 n-6) to the diet alters lipogenesis enhancing deposition of omega-6 fatty acids. This study examines changes in lipogenic gene expression of subcutaneous adipose tissue from eighteen steers fed increasing levels of dietary corn oil [0 (NONE), 0.31 kg/d (MED) and 0.62 kg/d (HI)] using two platforms, qPCR and microarray. The results show that MED level of oil supplementation up-regulates gene expression of key lipogenic enzymes but that as oil supplementation reaches HI level mRNA encoding lipogenic enzymes responsible for de novo synthesis and desaturation are down-regulated. Changes in specific lipogenic mRNA levels are correlated with changes in tissue fatty acid composition where de novo and desatured fatty acids were reduced with the highest level of oil supplementation.