Forages and pastures symposium: forage biodegradation: advances in ruminal microbial ecology.

The rumen microbial ecosystem provides ruminants a selective advantage, the ability to utilize forages, allowing them to flourish worldwide in various environments. For many years, our understanding of the ruminal microbial ecosystem was limited to understanding the microbes (usually only laboratory-amenable bacteria) grown in pure culture, meaning that much of our understanding of ruminal function remained a "black box." However, the ruminal degradation of plant cell walls is performed by a consortium of bacteria, archaea, protozoa, and fungi that produces a wide variety of carbohydrate-active enzymes (CAZymes) that are responsible for the catabolism of cellulose, hemicellulose, and pectin. The past 15 years have seen the development and implementation of numerous next-generation sequencing (NGS) approaches (e.g., pyrosequencing, Illumina, and shotgun sequencing), which have contributed significantly to a greater level of insight regarding the microbial ecology of ruminants fed a variety of forages. There has also been an increase in the utilization of liquid chromatography and mass spectrometry that revolutionized transcriptomic approaches, and further improvements in the measurement of fermentation intermediates and end products have advanced with metabolomics. These advanced NGS techniques along with other analytic approaches, such as metaproteomics, have been utilized to elucidate the specific role of microbial CAZymes in forage degradation. Other methods have provided new insights into dynamic changes in the ruminal microbial population fed different diets and how these changes impact the assortment of products presented to the host animal. As more omics-based data has accumulated on forage-fed ruminants, the sequence of events that occur during fiber colonization by the microbial consortium has become more apparent, with fungal populations and fibrolytic bacterial populations working in conjunction, as well as expanding understanding of the individual microbial contributions to degradation of plant cell walls and polysaccharide components. In the future, the ability to predict microbial population and enzymatic activity and end products will be able to support the development of dynamic predictive models of rumen forage degradation and fermentation. Consequently, it is imperative to understand the rumen's microbial population better to improve fiber degradation in ruminants and, thus, stimulate more sustainable production systems.

Forage degradation in the rumen is critical to producing ruminant animals. For many years, scientists were limited to biochemical techniques to understand how ruminal microbes degraded forage, impairing our understanding of which microbes were involved with degrading which forage components. However, we have understood that as the ruminant opened up plant cells to microbial activity, a succession of microbes was involved in colonizing and breaking fiber into increasingly smaller pieces. The recent development of sequencing techniques has allowed a more detailed understanding of changes in the microbial population of the rumen during forage degradation and the types of degradative enzymes produced by this complex microbial ecosystem. We described the enzymes involved in the degradation of specific forage components, how their end products impact the microbial population through cross-feeding interactions, and how fermentation products can impact food animal production.

Lipid metabolism mRNA expression and cellularity of intramuscular adipocytes within the Longissimus muscle of Angus- and Wagyu-sired cattle fed for a similar days on feed or body weight endpoint.

This study investigates intramuscular (IM) adipocyte development in the Longissimus muscle (LM) between Wagyu- and Angus-sired steers compared at a similar age and days on feed (D) endpoint or similar body weight (B) endpoint by measuring IM adipocyte cell area and lipid metabolism mRNA expression. Angus-sired steers (AN, n = 6) were compared with steers from two different Wagyu sires (WA), selected for either growth (G) or marbling (M), to be compared at a similar days on feed (DOF; 258 ± 26.7 d; WA-GD, n = 5 and WA-MD, n = 5) in Exp. 1 or body weight (BW; 613 ± 18.0 kg; WA-GB, n = 4 and WA-MB, n = 5) in Exp. 2, respectively. In Exp. 1, WA-MD steers had a greater (P ≤ 0.01) percentage of IM fat in the LM compared with AN and WA-GD steers. In Exp. 2, WA-MB steers had a greater (P ≤ 0.01) percentage of IM fat in the LM compared with AN and WA-GB steers. The distribution of IM adipocyte area was unimodal at all biopsy collections, with IM adipocyte area becoming progressively larger as cattle age (P ≤ 0.01) and BW increased (P ≤ 0.01). Peroxisome proliferator activated receptor delta (PPARd) was upregulated earlier for WA-MD and WA-MB cattle compared with other steers at a similar DOF and BW (P ≤ 0.02; treatment × biopsy interaction). Peroxisome proliferator activated receptor gamma was upregulated (PPARg) at a lesser BW for WA-MB steers (P = 0.09; treatment × biopsy interaction), while WA-MD steers had a greater (P ≤ 0.04) overall mean PPARg mRNA expression compared with other steers. Glycerol-3-phosphate acyltransferase, lipin 1, and hormone sensitive lipase demonstrated mRNA expression patterns similar to PPARg and PPARd or CCAAT enhancer binding protein beta, which emphasizes their importance in marbling development and growth. Additionally, WA-MD and WA-MB steers often had a greater early mRNA expression of fatty acid transporters (fatty acid transport protein 1; P < 0.02; treatment × biopsy interaction) and binding proteins (fatty acid binding protein 4) compared with other steers. Cattle with a greater marbling propensity appear to upregulate adipogenesis at a younger chronological and physiological maturity through PPARd, PPARg, and possibly adipogenic regulating compounds, lysophosphatidic acid, and diacylglycerol. These genes and compounds could be used as potential markers for marbling propensity of cattle in the future.

This study investigates intramuscular (IM) adipocyte development in the Longissimus muscle (LM) between Wagyu- and Angus-sired steers compared at a similar age and days on feed (D) or similar body weight (B) endpoint by measuring IM adipocyte cell area and lipid metabolism mRNA expression. Angus-sired steers (AN) were compared with steers from two different Wagyu sires (WA), selected for either growth (G) or marbling (M), to be compared at a similar days on feed (DOF; WA-GD, and WA-MD) in Exp. 1 or body weight (BW; WA-GB, and WA-MB) in Exp. 2. The WA-MD and WA-MB steers had a greater percentage of IM fat in the LM compared with AN and WA-GD and WA-GB steers. Intramuscular adipocyte cellularity analysis indicated few differences due to adipocyte size; therefore, marbling differences were likely due to adipocyte number. The WA-MD and WA-MB steers typically experienced an earlier or a greater mRNA upregulation for adipogenic genes that regulate fatty acid transport and triglyceride synthesis compared with other steers. The pattern of mRNA expression was very similar between steers compared at either a similar age and DOF or BW. Indicating that the timing of energy intake above maintenance requirements may be more influential than age for the upregulation of lipid metabolism in cattle.

Examination of weekly mammary parenchymal area by ultrasound, mammary mass, and composition in Holstein heifers reared on 1 of 3 diets from birth to 2 months of age.

Monitoring in vivo growth of mammary parenchyma (PAR) has historically been difficult, necessitating slaughter studies to measure PAR quantity. Advances in ultrasound (US) technology warrant revisiting its use as a noninvasive tool to monitor PAR growth in vivo. The level of nutrient intake during the first 2mo of life may affect measures of mammary growth and composition. Objectives were to examine the utility of US as an in vivo tool to quantify PAR cross-sectional area in Holstein heifers reared on 1 of 3 diets from birth to 2mo of age, assessing potential dietary effects; assess the relationships between weekly US measurements, teat length, manual palpation of PAR scores, and PAR mass at 2mo of age; and examine mammary composition in experimental animals. Holstein heifers (n=24; 41±1kg of initial body weight) from a single farm were randomly assigned to 1 of 3 milk replacers that differed in source and amount of fat. Milk replacer was fed at 660g of dry matter/d until weaning at 42 d. Heifers had ad libitum access to a common calf starter (20% crude protein) and water for the duration of the 56-d trial. Teat length and palpation scores were obtained weekly. A real-time B-mode US with a 7.5-MHz convex probe was used to examine 2-dimensional PAR area in all 4 glands of heifers once weekly from 2 to 3 d of age to harvest at 56 d. The left front and left rear glands were also examined by US 24h postharvest to validate final US measurements, and then bisected to produce a sagittal plane view of PAR for comparison with US images. Mass and composition of mammary gland tissue were determined at 8 wk using standard methodology. Over the course of this 8-wk trial, average teat length increased from 11 to 17mm. The PAR area started small (6.6±3.2mm(2) per gland) and increased to 42.1±2.5mm(2) per gland by the end of the trial. As anticipated, based on measurements obtained at slaughter, US measurements were more related to amount of PAR (r=0.74) than either teat length (r=0.34) or palpation scoring (r=0.63). Importantly, US is quantitative, whereas palpation scoring is subjective. Diet did not affect mass or composition of PAR in young heifers; total udder PAR mass averaged 1.40±0.80g. In conclusion, we showed that in heifers younger than 2mo of age, obtaining weekly PAR measurements via ultrasound is an effective quantitative tool for measuring changes in PAR area in vivo. Future studies may incorporate and expand upon the methods developed here to determine what quantitative evaluation of PAR in young heifers can reveal about milk production capacity.

Effects of sulfur and acidity on performance and digestibility in feedlot lambs fed dried distillers grains with solubles.

Negative interactions of the acidity and S content of dried distillers grains with solubles (DDGS) have not been quantified. The objectives of this study were to determine the effects of dietary S, dietary acidity, and their interactions on growth, feed efficiency, ruminal H2S concentration, and apparent nutrient digestibility in lambs fed DDGS-based diets. To neutralize acidity, the DDGS was untreated or treated with 2% NaOH. Dietary S content was adjusted with Na2SO4 to achieve a 0.2 percentage unit difference in dietary S. Experiment 1 included 72 ewe and wether lambs (BW = 24.9 ± 0.4 kg) penned by sex in 24 pens, blocked by BW, and allotted in a 2 × 2 factorial arrangement of treatments for a 60 d feeding trial. On a DM basis, diets were: i) 60% DDGS + 0% Na2SO4, ii) 60% DDGS + 0.88% Na2SO4, iii) 62% NaOH-treated DDGS + 0% Na2SO4, and iv) 62% NaOH-treated DDGS + 0.83% Na2SO4. There were no interactions (P ≥ 0.19) of dietary S concentration and NaOH treatment. Lambs fed DDGS treated with 2% NaOH had 4% greater (P = 0.05) final BW and tended (P ≤ 0.07) to have greater DMI and ADG than lambs fed untreated DDGS; however, NaOH treatment did not affect (P = 0.42) G:F. Increasing dietary S with Na2SO4 tended (P = 0.07) to reduce DMI; however, dietary S did not affect (P > 0.17) ADG, G:F, or final BW. Rumen H2S concentration in gas samples collected on d 32 and 60 were not (P > 0.17) affected by dietary S or NaOH treatment. Experiment 2 included 24 lambs (initial BW = 43.0 ± 4.4 kg) used to determine the effects of NaOH treatment of DDGS and sulfur (Na2SO4) supplementation on N retention and the apparent digestibility of NDF, ADF, and OM. Treatments were similar to those used in Exp. 1, except that dietary DDGS was reduced to 45% of DM. Intake of DM, NDF, ADF, and N was not affected (P > 0.16) by NaOH treatment of DDGS or dietary S (Na2SO4) content. Treating DDGS with 2% NaOH reduced (P = 0.05) NDF digestibility by 10% compared with lambs fed untreated DDGS. Urine pH for lambs consuming DDGS treated with 2% NaOH was increased (P = 0.01) 1.74 pH units compared with urine from lambs fed untreated DDGS. Nitrogen intake and retention were not affected (P > 0.35) by dietary S concentration or NaOH treatment of DDGS. For feedlot lambs, treating DDGS with 2% NaOH did not lower ruminal H2S concentrations, but tended to increase ADG and DMI. Although treating DDGS with 2% NaOH was an effective way to neutralize the acidity in DDGS, this treatment reduced NDF digestibility.

Prepartum dietary energy source fed to beef cows: II. Effects on progeny postnatal growth, glucose tolerance, and carcass composition.

Mature Angus-cross beef cows (n = 228) were used to evaluate effects of prepartum dietary energy source on postnatal growth and carcass composition of progeny in a 2-yr study. Starting at approximately 160 d of gestation, cows were fed diets consisting of 1 of 3 primary energy sources: grass hay (HY), corn (CN), or dried corn distillers grains with solubles (DG). The CN and DG diets were limit-fed to achieve similar energy intakes as cows fed HY. Following parturition, cows were fed a common diet and managed as a single group. Calves were weaned at an average of 185 ± 6 d of age and backgrounded for 28 d. A subset of progeny (n = 134) was individually fed a common finishing diet until slaughter, when each calf reached 1.2 ± 0.05 cm of backfat. A glucose tolerance test (GTT) was conducted in year 2 on 4 calves/treatment after 41 and 111 d on the finishing diet (DOF). Calf birth weights were greater (P = 0.002) in calves from cows fed CN and DG than calves from cows fed HY, and weaning BW (P = 0.08) was less for calves from cows fed HY vs. CN. Receiving BW, final BW, and HCW did not differ (P ≥ 0.16) among treatments. No difference (P ≥ 0.28) in ADG, morbidity, and mortality from birth to slaughter was observed among treatments. In response to a GTT, increased DOF resulted in greater (P ≤ 0.005) fasting insulin, faster glucose disappearance rate, and greater insulin:glucose area under the curve ratio. Glucose disappearance rate was greater (P = 0.01) in calves from cows fed CN than in calves from cows fed HY or DG. A greater initial insulin response (P = 0.005) was observed in calves from cows fed CN or DG than in calves from cows fed HY. Carcass traits used to measure yield grade did not differ (P ≥ 0.19) among treatments. Calves from dams fed CN had the lowest marbling score (P = 0.03) and intramuscular fat content (P = 0.07). These results indicate that prepartum maternal dietary energy source can alter fetal adipose tissue development and insulin sensitivity resulting in long-term effects on progeny's intramuscular fat deposition. Moreover, present findings suggest that increasing the number of days on a corn-based finishing diet increases insulin resistance in beef cattle.

Effects of copper supplementation on feedlot performance, carcass characteristics, and rumen sulfur metabolism of growing cattle fed diets containing 60% dried distillers grains.

The effects of 3 supplemental Cu concentrations on feedlot performance, mineral absorption, carcass characteristics, and ruminal S metabolism of cattle fed diets containing 60% dried distillers grains with solubles (DDGS) were evaluated in 2 experiments. Experiment 1 was conducted with 84 Angus-cross yearling steers and heifers (initial BW = 238 ± 36 kg), which were blocked by gender and allocated to 12 pens. Supplemental dietary Cu (tribasic copper chloride) treatments were: 1) 0 mg Cu/kg diet DM, 2) 100 mg Cu/kg diet DM, 3) 200 mg Cu/kg diet DM. The remainder of the diet was DDGS (60%), grass hay (10%), pelleted soy hulls (15%), and a vitamin-mineral supplement (15%). Diets were offered ad libitum throughout the finishing phase (168 d). Three cattle from each pen (n = 36) were harvested on d 168 and carcass data and liver samples were collected. Copper supplementation did not affect ADG (P = 0.22). However, the nonsignificant trend for increased ADG and decreased DMI led to a linear increase (P = 0.02) feed efficiency (G:F = 0.167, 0.177, and 0.177 for 0, 100, and 200 mg Cu/kg diet DM, respectively). The apparent absorption of Cu decreased quadratically (P = 0.07) and the apparent absorption of Mn and Zn were decreased linearly (P = 0.03 and P = 0.05, respectively) with increased Cu supplementation. Cattle supplemented with 100 or 200 mg Cu/kg diet DM had greater liver Cu concentrations (P < 0.01) than cattle that were not supplemented with Cu. There were no treatment effects (P > 0.10) on HCW, LM area, USDA yield grade, backfat, or marbling score. Experiment 2 was conducted with 6 ruminally fistulated steers that were fed the same diets as in Exp 1 in a replicated 3 × 3 Latin Square design. Copper supplementation did not affect (P > 0.10) ruminal pH or liquid S(2-) concentrations in steers consuming 60% DDGS diets (total dietary S = 0.55%). From 3 to 9 h after feeding, H(2)S gas concentration was decreased in those cattle supplemented with 100 mg Cu/kg diet. Concentration of H(2)S gas did not differ among cattle supplemented with 0 or 200 mg Cu/kg diet DM on 60% DDGS diets. Supplemental Cu improved feed efficiency in cattle consuming diets containing 60% DDGS; however, effects of Cu on rumen S metabolism were minimal even when supplemented at twice the maximum tolerable limit for beef cattle (NRC, 2000).

Effects of timing and duration of dietary vitamin A reduction on carcass quality of finishing beef cattle.

Two feedlot studies were conducted to investigate the timing and duration of supplemental vitamin A withdrawal from feedlot cattle (Bos taurus) diets to reduce intramuscular adipose tissue vitamin A concentration and improve carcass quality. In Exp. 1, Angus crossbred steers (n = 84, BW = 211 ± 4 kg) were allotted to 4 treatments: no supplemental vitamin A for 227 d, no supplemental vitamin A for 112 d followed by 115 d of supplemental vitamin A, supplemental vitamin A for 112 d followed by no supplemental vitamin A for 115 d, or supplemental vitamin A for 227 d. In Exp. 2, Angus crossbred steers (n = 80, BW = 210 ± 5 kg) were allotted to 4 treatments: early weaning with or without supplemental vitamin A, and traditional weaning with or without supplemental vitamin A. In both experiments, serum vitamin A concentrations were greatest (P < 0.05) 56 d after cattle were weaned and placed in the feedlot, regardless of feedlot dietary vitamin A concentration. Hepatic vitamin A stores were dramatically decreased (P < 0.05) in the first 56 d and remained depressed as long as steers were not supplemented with vitamin A. At the end of the finishing period, vitamin A concentrations were less in intramuscular than subcutaneous adipose tissue. Growth was not affected by finishing cattle without supplemental dietary vitamin A (P > 0.10). Dietary vitamin A supplementation did not affect USDA yield grades. However, in Exp. 2, cattle without supplemental vitamin A had greater (P < 0.001) ether extractable lipid in the LM. Ether extractable lipid in the LM or marbling scores were enhanced when intramuscular adipose tissue vitamin A concentration was reduced in response to feeding diets without supplemental vitamin A.

Effects of forage level in feedlot finishing diets on carcass characteristics and palatability of Jersey beef.

Jersey cattle are known for producing carcasses with a greater amount of marbling, but they require more days on feed to achieve acceptable market weights compared with other breeds. The objective of this study was to evaluate the effect of dietary forage (12 vs. 24% sudangrass:alfalfa hay, DM basis) in steam-flaked, corn-based finishing diets on carcass characteristics, beef palatability, and retail color stability of steaks from Jersey beef compared with conventionally fed commodity beef strip loins (COM) of identified quality (Choice(-) and Select(+)). Jersey steers (n = 77) were blocked by BW and randomly assigned to 1 of the following treatments for a 383-d trial period: Jersey low 12% (JL; n = 38) or Jersey high 24% (JH; n = 39) forage (DM basis). A comparison group was selected from conventionally fed cattle on the same day of slaughter as the Jersey treatments, and strip loins from USDA Select(+) (COM; n = 20) and Choice(-) (COM; n = 20) were removed for data analysis. Seventy-two hours postmortem, strip loins were removed, vacuum-packaged, and aged at 3°C for 18 d postmortem. After the aging period, steaks from the LM were sliced, vacuum-packaged, and frozen (-20°C) until analyzed. Jersey steaks had reduced (P < 0.05) Warner-Bratzler shear force values compared with COM steaks. Trained sensory panelists rated JL greater (P < 0.05) for initial and sustained tenderness and initial juiciness than COM, whereas JH was intermediate. As expected, marbling was greater (P < 0.05) for both JL and JH compared with COM, and trained sensory panel sustained juiciness, beef flavor intensity, and overall acceptability scores were greater (P < 0.05) for both JL and JH compared with COM; however, no differences (P = 0.14) were reported for consumer tenderness and flavor. Objective color (L*, a*, b*) measurements decreased (P < 0.05) over time across treatments. There were no differences among treatments for lightness (L*); however, overall during retail display JL were less (P < 0.05) red (a*) and yellow (b*) than JH and COM. Subjective color scores indicated both JL and JH were less red (P < 0.05) than COM. Steaks from Jersey were equal to and on some measurements more desirable than steaks from COM carcasses for both color stability and palatability. These results suggest that dietary forage level had minimal effects on carcass characteristics and beef palatability. However, feeding a low-forage diet decreases input cost and potentially results in a greater valued carcass. Finishing long-fed (383 d) Jersey steers can meet beef industry expectations with respect to quality grade.

Concentration of pro-vitamin A carotenoids in common beef cattle feedstuffs.

Quantification of the pro-vitamin A carotenoids in feedstuffs commonly fed to livestock has been ignored for many years. A greater dietary concentration of vitamin A has the potential to limit adipogenesis in cattle, thereby reducing carcass quality and value. A survey of 18 feedstuffs commonly fed to beef cattle was conducted for determination of vitamin A equivalents based on analysis of carotenoids. The pro-vitamin A carotenoids of interest were ß-carotene, α-carotene, and ß-cryptoxanthin. Collaborators in 5 states collected the feedstuffs and then shipped them to The Ohio State University for compilation and analysis. Carotenoids were extracted from the feedstuffs and then quantified using HPLC with photodiode array analysis. Fresh fescue pasture contained approximately 10 times more vitamin A equivalents than hay and 5 times more than corn silage (39,865, 2,750, and 6,900 IU of vitamin A/kg of DM for fresh pasture, hay, and corn silage, respectively). Beta-cryptoxanthin and α-carotene could not be detected in any forage samples. Hay and corn silage vitamin A equivalents decreased over extended periods of time from harvest to sample collection. Corn was the only feedstuff to have appreciable concentrations of all 3 pro-vitamin A carotenoids quantified. Corn processing had a minimal impact on the vitamin A equivalents. High-moisture corn contained 54% more vitamin A equivalents than whole shelled corn (378 and 174 IU of vitamin A/kg of DM, respectively). Pro-vitamin A carotenoids were more concentrated in corn coproducts than in whole shelled corn. The drying of distillers grains with solubles may significantly degrade ß-carotene (800 and 480 IU/kg of DM for wet and dry distillers grains, respectively). Soybean-based feedstuffs contain a small concentration of pro-vitamin A carotenoids, at 55 and 45 IU of vitamin A/kg of DM for soybean meal and soybean hulls, respectively. Overall, there was considerable variation in the pro-vitamin A content of feedstuffs based on location and storage conditions. An extensive analysis of feedstuffs would need to be conducted for an accurate estimation of the vitamin A content of feedlot cattle diets.

Restriction of vitamin A and D in beef cattle finishing diets on feedlot performance and adipose accretion.

Feedlot producers often exceed NRC recommendations for vitamin A and D supplementation; however, increased concentrations of these vitamins have been shown to limit adipocyte differentiation in vitro. A feedlot trial was conducted using 168 Angus crossbred steers (BW = 284 ± 0.4 kg) allotted to 24 pens. The experiment had a 2 × 2 factorial arrangement of treatments: no supplemental vitamin A or D (NAND), 3,750 IU vitamin A/kg dietary DM with no supplemental vitamin D (SAND), no supplemental vitamin A and 1,860 IU vitamin D/kg dietary DM (NASD), and 3,750 IU and 1,860 IU vitamin A and D/ kg dietary DM (SASD), respectively. Serum, liver, and intramuscular and subcutaneous adipose tissue retinol concentrations were decreased in (P < 0.001) in cattle fed the no supplemental vitamin A diets (NAND and NASD combined) compared with those consuming supplemental vitamin A (SAND and SASD combined) diets. In addition, intramuscular retinol concentration was 38% less than in the subcutaneous depot. Serum 25(OH)D(3) concentrations were reduced (P < 0.001) during the first 70 d when cattle were fed no supplemental vitamin D diets (NAND and SAND combined); however, liver 25(OH)D(3) concentrations remained unchanged (P > 0.10) through d 184. Serum and liver 25(OH)D(3) concentrations increased (P < 0.001) with vitamin D supplementation (NASD and SASD combined). The DMI, ADG, G:F, and morbidity were not affected (P > 0.10) by dietary concentration of vitamin A or D. There were vitamin A and D interactions (P < 0.03) for backfat thickness and USDA Yield grade. Cattle fed the NAND diet had greater (P < 0.03) Yield grades than other treatments because of greater (P < 0.005) 12th rib backfat thickness in NAND steers than the NASD and SAND steers. Vitamin D concentrations were attenuated and minimal carcass adiposity responses to vitamin D supplementation were observed. Feeding a diet without supplemental vitamin A increased (P < 0.05) Quality grades and marbling scores and tended (P = 0.06) to increase ether extractable lipid of the LM. As retinol and 25(OH)D(3) concentrations in feedlot cattle declined as a result of a lack of dietary supplementation, adipose accretion increased, resulting in elevated Quality and Yield grades. Withdrawal of supplemental vitamin A, D, or both from the finishing diet of feedlot beef cattle had minimal impact carcass composition.

Effects of diet and Aspergillus oryzae extract or Saccharomyces cervisiae on growth and carcass characteristics of lambs and steers fed to meet requirements of natural markets.

Two studies were conducted to determine the effects of diet and feed additive on growth and carcass characteristics of lambs and cattle destined for all natural markets. In Exp. 1, 48 Dorset × Hampshire lambs (initial BW 29.4 ± 0.1 kg) were used in a randomized complete block experiment to determine the effects of Aspergillus oryzae extract, Amaferm (AMF) supplementation (1 g/d) in an 85% concentrate diet on growth and carcass characteristics. Lambs were allotted to 12 pens (4 lambs per pen), and blocked by sex and BW. Lambs were fed until the average BW of each pen reached a target BW (55.4 kg for wethers and 50.0 kg for ewes), at which time the entire pen of lambs was slaughtered. Amaferm resulted in a greater (P=0.07) G:F. In Exp. 2, 168 crossbred steers (initial BW 300 ± 0.7 kg) were used in a trial with a 3 × 2 factorial arrangement of treatments to examine the effects of 0.5 g/d of Saccaromyces cervisiae boulardii CNCM 1079-Levucell SB (LEV), or 3 g/d of AMF with 2 corn sources, dry whole-shelled corn or high moisture corn, on growth and carcass characteristics. Neither LEV nor AMF improved (P>0.10) carcass characteristics compared with control or non-feed-supplemented steers. Addition of LEV to high-concentrate, corn-based diets did not improve (P>0.10) growth performance of feedlot steers. However, addition of AMF to a diet composed of dry whole-shelled corn resulted in an improvement (P<0.05) in G:F (0.208 vs. 0.194). Results indicate that at the amounts fed, AMF may improve G:F for lambs and steers fed dry corn-based finishing diets.

Winter-feeding systems for gestating sheep I. Effects on pre- and postpartum ewe performance and lamb progeny preweaning performance.

Mature pregnant crossbred ewes (n = 90) were used in a randomized complete block design and assigned to 1 of 3 winter-feeding systems differing in primary feed source: haylage (HL), limit-fed corn (CN), or limit-fed dried distillers grains (DDGS). Effects of these winter-feeding strategies on ewe and lamb performance were determined. Diets were formulated to meet or exceed NRC (1985) nutrient requirements during gestation and were fed from about d 60 of gestation until parturition. All ewes were fed a common diet postpartum. Every 2 wk during gestation, BW and BCS were collected and diets were adjusted to maintain similar BW gain for ewes fed CN and DDGS vs. HL. At 80 and 122 d of gestation, jugular blood samples were collected at 0, 3, 6, and 9 h postfeeding to measure plasma glucose, insulin, NEFA, and blood urea nitrogen concentrations. At birth, 6 lambs per treatment were killed to measure body composition. At 28 ± 2 d postpartum, milk yield was measured. Lambs were weaned at 61 ± 4 d of age. During mid gestation (d 60 to 115), BW gain of ewes was similar among treatments; however, at d 115 of gestation ewes fed HL had a smaller (P = 0.04) BCS than ewes fed DDGS or CN. Plasma glucose concentrations were greater (P ≤ 0.004) in ewes fed CN than in those fed HL or DDGS just before feeding on d 80 and 122 of gestation, whereas ewes fed DDGS vs. CN or HL had greater (P ≤ 0.04) plasma insulin concentrations at 3 h postfeeding. At parturition, ewe BW was greatest for DDGS, least for HL, and intermediate for CN (P ≤ 0.003). Ewes fed CN and DDGS had greater BCS at parturition than those fed HL, but by weaning, ewes fed DDGS had greater BCS (P ≤ 0.05) than those fed CN or HL. Birth BW tended (P = 0.09) to be heavier for lambs from ewes fed CN and DDGS than from those fed HL prepartum, but there was no difference (P = 0.19) due to ewe gestation diet on lamb BW at weaning. At birth, lamb muscle, bone, organ, and fat measures were not affected (P > 0.13) by treatment. Ewe milk production and lamb preweaning ADG were also similar (P > 0.44) among treatments. Prepartum dam winter feed source did not have detrimental effects on pre- or postpartum ewe performance, but altered prepartum maternal nutrient supply during gestation, which affected birth weight but not preweaning growth or mortality.

Winter-feeding systems for gestating sheep II. Effects on feedlot performance, glucose tolerance, and carcass composition of lamb progeny.

Mature pregnant crossbred ewes (n = 90) were used in a randomized complete block design experiment and were assigned to 1 of 3 winter-feeding systems differing in primary feed source: haylage (HL), limit-fed corn (CN), or limit-fed dried distillers grains (DDGS). Effects of these winter-feeding strategies on postweaning progeny performance were determined. Lamb progeny (n = 96) were weaned at 61 ± 4 d of age and fed a common high-concentrate diet. Lambs were assigned to feedlot pen (n = 18) based on dam mid-gestation pen. Growth rate, DMI, and ADG were determined for the first 40 d of the finishing period. At 96 ± 4 d of age, 1 wether lamb was randomly selected from each pen (n = 18) for a glucose tolerance test. The experiment was terminated, and lambs were slaughtered individually when they were determined to have achieved 0.6-cm 12th-rib fat thickness. After a 24-h chill, carcass data were collected and a 2.54-cm chop was removed from each lamb from the LM posterior to the 12th rib for ether extract analysis. Additional carcass measurements of bone, muscle, and fat from the shoulder, rack, loin, and leg were collected on 35 carcasses. At weaning, lamb BW was not different among treatments, whereas final BW tended to be greater (P = 0.09) for lambs from ewes fed DDGS and CN during gestation than from those fed HL. Overall lamb growth rate from birth to slaughter was not different among treatments. Lambs from ewes fed DDGS vs. CN or HL tended to have a greater initial insulin response (P = 0.09). Dressing percent was less (P = 0.04) in lambs from ewes fed DDGS, but no difference (P = 0.16) was detected in HCW among treatments. As expected, 12th rib fat thickness was similar among treatments, whereas LM area was largest to smallest (P = 0.05) in lambs from ewes fed CN, HL, and DDGS, respectively. Proportion of internal fat tended to be greatest to smallest (P = 0.06) in lambs from ewes fed DDGS, CN, and HL, respectively. Calculated boneless trimmed retail cuts percentage was less (P = 0.04) in lambs from ewes fed DDGS than CN or HL. Loin muscle weight as a percentage of wholesale cut tended (P = 0.10) to be greater in lambs from ewes fed CN and HL than DDGS, whereas other muscle, bone, and fat weights and proportions were similar (P > 0.24) among treatments. Prepartum diet during mid to late gestation of ewes altered postnatal fat and muscle deposition and may be associated with alterations in insulin sensitivity of progeny.

Adipogenic differentiation state-specific gene expression as related to bovine carcass adiposity.

Genetic regulation of the site of fat deposition is not well defined. The objective of this study was to investigate adipogenic differentiation state-specific gene expression in feedlot cattle (>75% Angus; <25% Simmental parentage) of varying adipose accretion patterns. Four groups of 4 steers were selected via ultrasound for the following adipose tissue characteristics: low subcutaneous-low intramuscular (LSQ-LIM), low subcutaneous-high intramuscular (LSQ-HIM), high subcutaneous-low intramuscular (HSQ-LIM), and high subcutaneous-high intramuscular (HSQ-HIM). Adipose tissue from the subcutaneous (SQ) and intramuscular (IM) depots was collected at slaughter. The relative expression of adipogenic genes was evaluated using quantitative PCR. Data were analyzed using the mixed model of SAS, and gene expression data were analyzed using covariate analysis with ribosomal protein L19 as the covariate. No interactions (P > 0.10) were observed between IM and SQ adipose tissue depots for any of the variables measured. Therefore, only the main effects of high and low accretion within a depot and the effects of depot are reported. Steers with LIM had smaller mean diameter IM adipocytes (P < 0.001) than HIM steers. Steers with HSQ had larger mean diameter SQ adipocytes (P < 0.001) than LSQ. However, there were no differences (P > 0.10) in any of the genes measured due to high or low adipose accretion. Preadipogenic delta-like kinase1 mRNA was greater in the IM than the SQ adipose tissue; conversely, differentiating and adipogenic genes, lipoprotein lipase, PPARγ, fatty acid synthetase, and fatty acid binding protein 4 were greater (P < 0.001) in the SQ than the IM depot. Intramuscular adipocytes were smaller than SQ adipocytes and had greater expression of the preadipogenic gene, indicating that more hyperplasia was occurring. Meanwhile, SQ adipose tissue contained much larger (P < 0.001) adipocytes that had a greater expression (P < 0.001) of differentiating and adipogenic genes than did the IM adipose tissue, indicating more cells were undergoing differentiation and hypertrophy. Adipogenic differentiation state-specific gene expression was not different in cattle with various phenotypes, but adipogenesis in the SQ and IM adipose tissues seems to occur independently.

Prepartum dietary energy source fed to beef cows: I. Effects on pre- and postpartum cow performance.

Mature Angus-cross beef cows (n = 144) were used to determine effects of late gestation dietary energy source on pre- and postpartum cow performance in a complete randomized block design experiment. Cows were adapted to diets starting at 167 +/- 9 d of gestation and fed until 1 wk before expected calving date. Cows were fed 1 of 3 dietary energy sources: grass hay (HY), corn (CN), or dried distillers grains (DDGS). Cows allotted to HY were allowed ad libitum access to round-bale grass hay, and average hay disappearance was 12.4 kg/d. Limit-fed corn and DDGS diets contained 5.3 kg of whole-shelled corn or 4.1 kg of DDGS, respectively, plus 2.1 kg of hay, and 1.0 kg of supplement to meet cow nutritional needs during late gestation and to allow for an energy intake similar to HY. Every 21 d, BW, BCS, and ultrasound measurement of backfat between the 12th and 13th ribs were collected. At 210 d in gestation, jugular blood samples were collected from cows at 0, 3, 6, and 9 h postfeeding and were analyzed for glucose, insulin, NEFA, and blood urea N (BUN) concentrations. After parturition, cows were fed a common diet and managed similarly. Milk production was determined by weigh-suckle-weigh procedure on d 31, 100, and 176 postpartum. Cows fed DDGS during late gestation gained more (P = 0.04) BW than cows fed HY or CN; however, no difference in BCS change was detected (P = 0.28) among treatments. Plasma glucose concentrations were similar among treatments (P = 0.64), whereas insulin concentrations at 3 h postfeeding were greater (P = 0.002) for cows fed DDGS than those fed HY or CN. Plasma BUN concentrations were greater (P < or = 0.02) for cows fed DDGS vs. CN or HY up to 6 h postfeeding. Birth weight was greater (P < 0.001) for calves from cows fed CN and DDGS than for those fed HY, but this did not result in any differences in frequency of dystocia (P = 0.21). Prepartum energy source did not affect conception rates (P = 0.79), milk production (P > or = 0.51), or milk composition (P > or = 0.39). Maternal dietary energy source in late gestation did not affect pre- or postpartum cow performance, but did change plasma hormones and metabolites during gestation. Heavier birth weights in calves from cows fed CN or DDGS indicate the changes in maternal metabolism affected energy partitioning of nutrients to the fetus and subsequent fetal growth.

Effects of dietary polyunsaturated fatty acid supplementation on fatty acid composition in muscle and subcutaneous adipose tissue of lambs.

Lambs (n = 48) were used in a 2 x 2 factorial arrangement of treatments to evaluate effects of inclusion of oil containing PUFA in high-concentrate diets (with or without) and duration of oil supplementation (pre- vs. postweaning) on CLA concentration of muscle and adipose tissue. Lambs were fed preweaning creep diets (with or without oil) corresponding to the dietary lactation treatment diet (with or without oil) of the dam. Dams blocked by lambing date and rearing type were randomly assigned to 1 of 2 lactation dietary treatments with or without oil supplementation. Creep diets contained approximately 70% concentrate and 30% roughage and were provided to lambs for ad libitum intake. At weaning (58.7 +/- 2.5 d of age), lambs (n = 48) were randomly assigned within preweaning treatment groups to 1 of 2 postweaning dietary treatments (with or without oil) and 16 pens in a randomized block design, blocked by sex and BW. Postweaning diets were formulated to contain approximately 80% concentrate and 20% roughage and were fed once daily for ad libitum intake. Soybean and linseed oil (2:1, respectively) replaced ground corn and provided 3% additional fat in pre- and postweaning diets. Lambs were slaughtered at 60.3 +/- 4.2 kg of BW. A subcutaneous fat (SQ) sample was obtained within 1 h postmortem and a LM sample at the 12th rib was obtained 24 h postmortem, and both were analyzed for fatty acid profile. Feedlot performance and carcass measurements were not affected (P >or= 0.26) by oil supplementation. Total CLA content of LM and SQ was not affected (P >or= 0.08) by oil supplementation pre- or postweaning, but trans-10, cis-12 CLA was greater (P = 0.02) in SQ from lambs supplemented with oil postweaning. Total PUFA content in LM was greater (P = 0.02) in lambs supplemented with oil pre- or postweaning as a result of increased concentrations of 18:2cis-9, cis-12 and longer chain PUFA. Conversely, pre- and postweaning oil supplementation resulted in less (P = 0.04) MUFA content in LM. Only postweaning oil supplementation increased (P = 0.001) SQ PUFA content. Feeding oils containing PUFA to lambs pre- and postweaning did not increase CLA content of muscle, whereas postweaning oil supplementation minimally increased CLA concentration of SQ fat. Inclusion of soybean and linseed oil in pre- and postweaning diets increased total PUFA content of SQ fat and muscle tissue without adversely affecting growth performance or carcass characteristics.

Effect of Wagyu- versus Angus-sired calves on feedlot performance, carcass characteristics, and tenderness.

Wagyu-sired (n = 20) and Angus-sired (n = 19) steers and heifers were used to compare the effects of sire breed on feedlot performance, carcass characteristics, and meat tenderness. Calves were weaned at 138 +/- 5 d of age and individually fed a finishing diet consisting of 65% whole corn, 20% protein/vitamin/mineral supplement, and 15% corn silage on a DM basis. Heifers and steers were slaughtered at 535 and 560 kg of BW, respectively. Carcasses were ribbed between the 12th and 13th (USDA grading system) and the 6th and 7th ribs (Japanese grading system) to measure fat thickness, LM area (LMA), and intramuscular fat (IMF). Two steaks were removed from the 12th rib location and aged for 72 h and 14 d to determine Warner-Bratzler shear force and cooking loss. Sire breed x sex interactions were not significant (P > 0.05). Angus-sired calves had greater (P < 0.05) ADG and DMI than Wagyu. Wagyu-sired calves had improved (P < 0.05) feed efficiency than Angus. Sire breed did not affect (P > 0.20) HCW, 12th-rib fat, or USDA yield grade. Carcasses of Wagyu had greater (P = 0.0001) marbling scores at the 12th rib than those of Angus (770.9 vs. 597.3 +/- 41.01, respectively). Carcasses of Wagyu also had greater (P < 0.02) 12th-rib IMF and 6th-rib IMF than Angus, resulting in a greater proportion of carcasses grading Prime (65.0 vs. 21.1%; P = 0.006). Carcasses from Wagyu tended (P = 0.08) to have greater LMA at the 12th rib, whereas Angus carcasses had greater (P < 0.05) LMA at the 6th rib. Steaks from Angus and Wagyu had similar (P > 0.50) tenderness at aging times of 72 h and 14 d. Cooking loss was greater (P < 0.01) for Angus than Wagyu steaks at 72 h and 14 d. Using Wagyu sires vs. Angus sires on British-based commercial cows combined with early weaning management strategies has the potential to produce a product with greater marbling, but is unlikely to significantly enhance tenderness.

Technical note: Occurrence in fecal microbiota of genes conferring resistance to both macrolide-lincosamide-streptogramin B and tetracyclines concomitant with feeding of beef cattle with tylosin.

Development of antimicrobial resistance in food animals receiving antimicrobials has been well documented among bacterial isolates, especially pathogens, but information on development of antimicrobial resistance at the microbial community level during long-term feeding of antimicrobials is lacking. The objective of this study was to examine the association between inclusion of tylosin in feed and occurrence of resistance to macrolide-lincosamide-streptogramin B (MLS(B)) in the entire fecal microbial communities of beef cattle over a feeding study of 168 d. A completely randomized design included 6 pens housed together in 1 barn, with each pen housing 10 to 11 steers. The control and tylosin groups each had 3 pens, with the former receiving no antimicrobial whereas the latter received both tylosin and monensin (11 and 29.9 mg/ kg of feed, respectively, DM) in feed. The abundance of genes conferring resistance to MLS(B) (erm genes) and tetracyclines (tet genes) were quantified using class-specific, real-time PCR assays. The abundances of erm and tet genes were analyzed with pens as experimental units using the MIXED procedure of SAS. Correlations between abundance of different resistance genes were calculated using the CORR procedure of SAS. We identified 4 classes (B, F, T, and X) of erm genes in fresh fecal samples collected at wk 2, 17, and 21 of feeding. From wk 2 to 17, the abundance of erm(T) and erm(X) increased (P < 0.05), whereas that of erm(B) and erm(F) did not. The abundance of the erm genes did not further change from wk 17 to 21. The tet(A/C), tet(G), and tet gene variants encoding ribosomal protection proteins (including classes M, O, P, Q, S, T, and W) appeared to be co-selected by tylosin feeding. Such co-selection of multiresistance at community level by one antimicrobial drug used in animals has the important implication that future studies should examine resistance to not only the antimicrobials used in animals, but also other antimicrobials, especially those used in human medicine, to fully assess the potential risk associated with antimicrobial use in animals. Both the erm and tet genes appeared to be disseminated among the microbial populations in all steers housed together.

Effect of vitamin A restriction on carcass characteristics and immune status of beef steers.

Sixty-eight Angus-based steers (224 +/- 7.6 kg of BW) were used to evaluate the effects of a prolonged dietary vitamin A restriction on marbling and immunocompetency. Steers were allotted randomly to 1 of 2 treatments: LOW (no supplemental vitamin A) and HIGH (diet supplemented with 2,200 IU of vitamin A/kg of DM). Diets contained 60% high-moisture corn, 20% roasted soybeans, 10% corn silage, and 10% of a protein supplement. Steers were penned and fed individually. For the first 141 d, steers were program-fed to achieve a gain of 1.1 kg/d. The last 75 d of the experiment, steers were offered feed for ad libitum intake. At slaughter, serum and liver samples were taken to determine their retinol content. To evaluate immunocompetency, 10 steers per treatment were selected randomly on d 141 and received an ovalbumen vaccine, and 21 d later, the steers were revaccinated. On d 182, blood samples were taken from the vaccinated steers to determine serum antibody titers by ELISA. Steers were slaughtered after 216 d on feed. Carcass characteristics were determined, and LM samples were taken for composition analysis. Subcutaneous fat samples were taken for fatty acid composition analysis. Performance (ADG, DMI, and G:F) was not affected by vitamin A restriction (all P > 0.10). Hot carcass weight, 12th-rib fat, and yield grade did not differ between LOW and HIGH steers (all P > 0.10). Marbling score (LOW = 574 vs. HIGH = 568, P = 0.79) and i.m. fat (LOW = 5.0 vs. HIGH = 4.7% ether-extractable fat, P = 0.57) were not increased by vitamin A restriction. Serum (LOW = 18.7 vs. HIGH = 35.7 mug/dL, P < 0.01) and liver (LOW = 6.3 vs. HIGH = 38.1 mug/g, P < 0.01) retinol levels were lower in LOW steers compared with HIGH steers at slaughter. Response to ovalbumin vaccination was not affected by vitamin A restriction (LOW = 13.1 vs. HIGH = 12.8 log(2) titers, P = 0.60). Slight changes in the fatty acid profile of s.c. fat of the steers were detected. A greater proportion of MUFA (LOW = 41.7 vs. HIGH = 39.9%, P = 0.03) and fewer SFA (LOW = 47.1 vs. 48.7, P = 0.03) were observed in vitamin A-restricted steers. This suggests that vitamin A restriction may affect the activity of desaturase enzyme (desaturase activity index, LOW = 46.9 vs. HIGH = 44.9, P = 0.01). Feeding a low vitamin A diet for 216 d to Angus-based steers did not affect performance, marbling score, or animal health and immunocompetency. Slight changes in the fatty acid profile of s.c. fat were observed, suggesting that vitamin A restriction may have affected desaturase enzyme activity.

Effect of low vitamin A diets with high-moisture or dry corn on marbling and adipose tissue fatty acid composition of beef steers.

Angus-cross steers (n = 165; 295 +/- 16 kg of BW) were used evaluate the effect of low vitamin A diets with high-moisture corn (HMC) or dry corn (DC) on marbling and fatty acid composition. Steers were allotted to 24 pens (7 steers/pen), such that each pen had the same average initial BW. Treatments were randomly allotted to the pens. The experiment had a completely randomized design, with a 2 x 2 factorial arrangement of treatments: low vitamin A (Lo, no supplemental vitamin A) and HMC (LoHMC); LoDC; high vitamin A (Hi, supplemented with 2,200 IU of vitamin A/kg of DM) and HMC (HiHMC); and HiDC. Diets contained 76% corn, 10% corn silage, 11% protein supplement, and 3% soybean oil (DM basis). Samples of feed ingredients were collected for carotenoid analysis. Blood samples were collected for serum retinol determination. Steers were slaughtered after 145 d on feed. Carcass characteristics and LM composition were determined. Samples from the s.c. fat depot were analyzed for fatty acid composition. High-moisture corn had a greater vitamin A content, based on its carotenoid content, than DC (614 vs. 366 IU/kg of DM, P < 0.01). No vitamin A x corn type interactions were detected for feedlot performance, carcass characteristics, or serum, s.c. fat, or liver retinol concentration. Average daily gain, DMI, and G:F were not affected by vitamin A (P > 0.05). Marbling score and USDA quality grade were greater (P < 0.05) in Lo vs. Hi steers. Hot carcass weight, backfat, and yield grade were not affected by the treatments (P > 0.05). Vitamin A and corn type did not affect LM composition (DM, ash, CP, or ether-extractable fat, P > 0.05). Vitamin A supplementation increased (P < 0.06) serum retinol on d 112 and 145 and increased (P < 0.01) liver retinol at slaughter (Lo = 38.7 vs. Hi = 102.9 mug/g). The s.c. fat retinol concentrations were less (P < 0.01) for Lo (0.8 mug/g) than for Hi (1.4 mug/g) at slaughter. Cell diameter of adipocytes in the i.m. depot was not affected by dietary vitamin A (P > 0.05). A vitamin A x corn type interaction was observed (P < 0.05) for the s.c. fat cellularity. Feeding HMC increased the number of cells per square millimeter when Lo diets were fed (LoHMC = 128 vs. LoDC = 100 cells/mm(2), P < 0.05), but not when Hi diets were fed (HiHMC = 109 vs. HiDC = 111 cells/mm(2), P > 0.05). The CLA content of adipose tissue was not affected by the treatments. Regardless of the corn type used, feeding low vitamin A diets for 145 d to Angus-cross steers increased marbling and quality grade without affecting yield grade, animal health, or performance.