Macrolide-susceptible probiotic Enterococcus faecium ST296 exhibits faecal-environmental-oral microbial community cycling among beef cattle in feedlots.

Enterococci are included in the United States National Antimicrobial Resistance Monitoring System to track antibiotic resistance among commensal Gram-positive enteric bacteria, largely due to their high abundance in food animals and in retail meat. In the U.S. cattle industry, macrolides are used to prevent and control liver abscesses, which cause significant economic losses. Previous studies have suggested that feeding tylosin and the intensity of the pen environment, both expand and sustain respectively the prevalence of multidrug resistance among enterococci in feedlot cattle. This has led to research into alternative feed supplements and improved stewardship practices. In a randomized controlled trial, we measured the impact of a probiotic and an altered pen environment on antimicrobial resistance among faecal Enterococcus spp. in cattle fed tylosin. Supplementing cattle with an Enterococcus faecium and Saccharomyces cerevisiae-based probiotic yielded the isolation of E. faecium of the probiotic sequence type (ST296) from faecal and environmental samples in treatment groups, as well as from cattle and the manure pack in nearby pens. Of importance, the probiotic strain also was found in a desiccated and milled manure pack sample taken 120 days after the initial trial ended. Phylogenetic and SNP analyses revealed clonal survival and spread compatible with faecal-environmental-oral recycling of the probiotic strain within and among cattle and pens. The increase in prevalence of the ST296 strain occurred concomitant with a decrease in ST240, the dominant sequence type associated with ermB and tet(M) resistance genes in this trial. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrate that a macrolide-susceptible probiotic Enterococcus faecium ST296 strain fed to beef cattle becomes fully embedded in the microbial community cycling of bacteria via faecal-environmental-oral transmission within and among feedlot pens. An initial investment in feeding the probiotic is thereby leveraged into expanding numbers of susceptible bacteria in cattle and their environment, even among those cattle fed tylosin.

Effect of monensin withdrawal on intake, digestion, and ruminal fermentation parameters by Bos taurus indicus and Bos taurus taurus steers consuming bermudagrass hay.

Effects of monensin withdrawal and cattle subspecies on the utilization of bermudagrass hay (14.3% CP, 72.3% NDF, and 36.9% ADF) were evaluated using ruminally cannulated steers (5 [BI] and 5 [BT]). Subspecies were concurrently subjected to a 2-period, 2-treatment crossover design. Treatments consisted of either 0 mg·steer·d-1 monensin with no previous monensin feeding (CON) or withdrawal from 200 mg·steer·d-1 monensin (MON) fed individually in 0.91 kg dried distillers' grains with solubles for 42 d. Withdrawal was evaluated for a 28-d period. Ruminal fluid was collected 2 h after feeding on d 0, 1, 4, 7, 14, and 21 after withdrawal for determination of pH, VFA, ruminal NH-N (RAN), rate of NH production, and CH production rate. Hay, ort, and fecal grab samples were collected d 23 through 28 after withdrawal for determination of intake and digestion. No subspecies × monensin, subspecies × day, or subspecies × monensin × day interactions were observed ( ≥ 0.11). An effect of day after monensin withdrawal was observed ( < 0.01) for total VFA concentration, with an increase following withdrawal followed by a decrease and then stabilization. Monensin × day after monensin withdrawal interactions ( ≤ 0.01) were observed for the acetate:propionate (A:P) ratio and molar percent of acetate and propionate. There was a decrease in molar percent of propionate between d 1 and 4 from 19.1 to 18.0; however, it remained greater ( ≤ 0.10) for MON than CON through d 7. Withdrawal increased molar percent of acetate from 68.3 to 69.8 between d 0 and 4 for MON steers. The A:P ratio was less ( ≤ 0.01) on d 0 for MON than for CON (3.4 vs. 4.0), but by d 4, it increased to 3.8 and was not different ( = 0.14) from CON. By d 14, no differences ( ≥ 0.88) remained for acetate, propionate, or the A:P ratio. After monensin withdrawal, monensin reduced ( < 0.01) RAN by 12.3% (2.09 vs. 1.83 m for CON and MON, respectively). Monensin withdrawal and cattle subspecies had no effect ( ≥ 0.23) on rate of NH production or CH production rate. Monensin withdrawal had no effect ( ≥ 0.45) on intake or digestibility parameters. Greater forage OM intake ( = 0.09; 21.2 vs. 19.2 g/kg BW) and OM digestibility ( < 0.01; 72.4 vs. 63.0%) resulted in greater ( < 0.01) total digestible OM intake (16.8 vs. 13.2 g/kg BW) in BT steers than in BI steers. These results suggest that BT steers are better able to utilize bermudagrass hay than BI steers. Upon monensin withdrawal, steers previously fed monensin continue to have a reduced A:P ratio for at least 7 d.

Effect of monensin inclusion on intake, digestion, and ruminal fermentation parameters by Bos taurus indicus and Bos taurus taurus steers consuming bermudagrass hay.

Effects of monensin inclusion and cattle subspecies on utilization of bermudagrass hay (13.7% CP, 77.3% NDF, and 38.8% ADF) were evaluated using ruminally cannulated steers (5 [BI] and 5 [BT]; 398 kg BW). Subspecies were concurrently subjected to a 2-period, 2-treatment crossover design. Treatments were 0 (CON) or 200 mg·steer·d monensin (MON) in 0.91 kg dried distillers' grains with solubles. Periods were 70 d in length: 20 d of adaptation, 22 d of sample collection, and 28 d for withdrawal of treatment. Steers were group housed during adaptation and moved to individual covered pens for sampling. Hay, ort, and fecal grab samples were collected d 21 through 25 for determination of intake and digestion. Ruminal fluid was collected with a suction strainer 0, 2, 4, 8, and 12 h after feeding on d 42 for pH, VFA, and ruminal NH-N (RAN) analysis. Additionally, at h 2, ruminal fluid and contents were collected for determination of rate of NH production and CH production rate. No subspecies × monensin interactions were observed ( ≥ 0.12). Monensin had no effect ( ≥ 0.16) on intake or digestibility parameters. No subspecies effect ( ≥ 0.11) was observed for forage OM intake, total OM intake, or OM digestion. Total digestible OM intake tended to be greater ( = 0.06) for BT steers than for BI steers (14.0 vs. 12.2 g/kg BW). There was an effect of hour after feeding ( ≤ 0.01) on pH, total VFA, acetate:propionate ratio, and molar percent acetate and propionate. Total VFA concentration was greater ( = 0.01) in CON steers than in MON steers (66.5 vs. 62.0 m). Monensin decreased molar percent acetate ( = 0.02) from 72.5 to 71.2% and increased molar percent propionate ( < 0.01) from 16.9 to 18.7%, resulting in a reduced ( < 0.01) acetate:propionate ratio (from 4.34 to 3.85). Although not significantly ( = 0.19), monensin numerically reduced the CH production rate by 15.8%. Greater ( = 0.07) CH production rate tended to be observed in BI steers than in BT steers (21.4 vs. 16.6 µmol CH·mL·h, respectively). Monensin had no effect ( ≥ 0.32) on pH, RAN, or rate of NH production. A subspecies × hour after feeding interaction was observed for RAN, with BT having greater RAN at h 0 and 4, whereas BI had greater RAN at h 2, 8, and 12. Overall, monensin decreased the acetate:propionate ratio and total VFA concentration but had no effect on forage utilization. steers consumed less digestible OM and had a greater CH production rate compared with BT steers, suggesting BT were better able to utilize the available forage resource than BI.

Effect of source and level of protein supplementation on rice straw utilization by Brahman steers.

Seven ruminally cannulated Brahman steers were used in a 7 × 4 incomplete block design to determine the effects of cottonseed meal (CSM; 43.9% CP, 82.9% RDP) or dried distillers' grains (DDG; 27.5% CP, 43.6% RDP) supplementation on rice straw utilization (47 g/kg CP and 681 g/kg NDF). Treatments consisted of a negative control receiving no supplement (control) and 3 levels (60, 120, and 180 mg N/kg BW) of either CSM or DDG. Periods were 14 d with 8 d for adaptation and 6 d for data collection. Steers had ad libitum access to rice straw and were fed supplements daily. Increased supplementation resulted in a linear increase ( ≤ 0.06) in forage OM intake from 13.5 g/kg BW by controls to 15.5 and 16.1 g/kg BW for 180 mg N/kg BW of DDG and CSM, respectively. No differences between sources were observed ( = 0.84). Total digestible OM intake was increased by supplementation (linear, < 0.01) from 6.9 g/kg BW (control) to 10.0 and 11.2 g/kg BW for 180 mg N/kg BW of CSM and DDG, respectively. A greater response was observed for DDG ( = 0.05) due to greater provision of supplement (g DM/d) to achieve isonitrogenous treatment levels. Total tract OM digestion tended to increase with DDG supplementation (linear, = 0.08) but not CSM supplementation ( = 0.19). Both supplements did not affect NDF digestion ( > 0.40) or calculated forage NDF digestibility ( > 0.40). Ruminal ammonia concentrations peaked 4 h after supplementation/feeding with the greatest concentration (4.0 m) observed for 180 mg N/kg BW of CSM and the lowest concentration at 4 h observed in the control (0.8 m). Provision of CSM resulted in a linear increase ( < 0.01) in average ruminal ammonia, in contrast to the quadratic response ( = 0.02) observed with DDG supplementation. Total VFA production linearly increased for both CSM and DDG supplementation ( = 0.09 and = 0.01, respectively). Protein supplements containing high and low levels of RDP were effective at improving intake and utilization of rice straw by Brahman steers.

Effect of postextraction algal residue supplementation on the ruminal microbiome of steers consuming low-quality forage.

Cattle consuming low-quality forages (LQF) require protein supplementation to increase forage utilization via ruminal fermentation. Biofuel production from algal biomass results in large quantities of postextraction algal residue (PEAR), which has the potential to elicit LQF utilization responses similar to cottonseed meal (CSM); however, its effect on ruminal bacterial communities is unknown. Five ruminally and duodenally cannulated Angus steers in a 5 × 5 Latin square had ad libitum access to oat straw diets. Treatments were infused ruminally and consisted of an unsupplemented control; PEAR at 50, 100, and 150 mg N/kg BW; and CSM at 100 mg N/kg BW. Ruminal samples were collected 4 h after supplementation on d 14 of each period and separated into solid and liquid fractions. Each sample was extracted for genomic DNA, PCR amplified for the V4 to V6 region of the 16S rRNA, sequenced on the 454 Roche pyrosequencing platform, and analyzed using the QIIME pipeline. Weighted UniFrac analysis and Morisita-Horn index demonstrated different community composition between liquid and solid fractions. Measures of richness including observed operational taxonomic units (OTU) and abundance coverage estimator metric decreased with greater PEAR provision (P ≤ 0.09). There were 42 core microbiome OTU observed in all solid fraction samples while the liquid fraction samples contained 30 core OTU. Bacteroidetes was the predominant phylum followed by Firmicutes in both fractions, which together characterized more than 90% of sequences. Relative abundance of Firmicutes increased with PEAR supplementation in the liquid fraction (linear, P = 0.02). Among Firmicutes, Lachnospiraceae, Ruminococcaceae, and Clostridiaceae families increased in the liquid fraction with greater PEAR supplementation (linear, P ≤ 0.03). Prevotella represented over 25% of sequences in all treatments, and relative abundance decreased in the solid fraction with increasing PEAR provision (linear, P = 0.01). Fibrobacter and Treponema decreased in the liquid fraction with increasing PEAR (linear, P < 0.10). Results suggest PEAR supplementation increased forage utilization by increasing members of Firmicutes within the liquid fraction of the rumen microbiome.

Effect of increasing amounts of postextraction algal residue on straw utilization in steers.

Algal biomass has been identified as a third-generation biofuel. Significant quantities of the coproduct postextraction algal residue (PEAR) remain after lipid extraction. After extraction, PEAR is concentrated in protein (17.9% CP on a DM basis and 32.5% CP on an ash-free basis), suggesting it may be an alternative to cottonseed meal (CSM) as a protein supplement. Our objectives were to determine the optimal level of PEAR supplementation to steers consuming straw and to compare the effects of PEAR supplementation on straw utilization and N metabolism with an isonitrogenous level of CSM. Five steers (198.2 ± 6.1 kg of BW), in a 5 × 5 Latin square, had ad libitum access to oat straw (80% NDF and 4.5% CP on a DM basis). Treatments were infused ruminally once daily and included no supplemental protein (CON); PEAR at 50, 100, and 150 mg N/kg BW; and CSM at 100 mg N/kg BW. Provision of PEAR increased total digestible OM intake (TDOMI) quadratically (P = 0.01) from 0.9 (CON) to 1.6 kg/d (100 mg N/kg BW of PEAR). Organic matter digestibility (OMD) increased quadratically (P < 0.01) with supplementation and was maximized (55% OMD) at 50 mg N/kg BW of PEAR. At isonitrogenous levels of PEAR and CSM, TDOMI was similar (P = 0.13) as was OMD (P = 0.50). Negative N balance was observed for all treatments except PEAR provided at 100 or 150 mg of N/kg BW. Nitrogen balance was quadratic (P < 0.01) with the greatest retention (1.84 g N/d) occurring at 100 mg N/kg BW of PEAR. There were no differences (P ≥ 0.22) between isonitrogenous PEAR and CSM supplementation in measurements of ruminal ammonia or VFA concentrations. Straw utilization was maximized when PEAR was provided at 100 mg N/kg BW. Our observations suggest cattle provided PEAR utilize straw in a manner similar to those supplemented CSM, indicating PEAR has potential to substitute for CSM as a protein supplement in forage-based operations.

Fatty acid biosynthesis and lipogenic enzyme activities in subcutaneous adipose tissue of feedlot steers fed supplementary palm oil or soybean oil.

We hypothesized that supplementing finishing diets with palm oil would promote adipocyte differentiation in subcutaneous adipose tissue of feedlot steers, and that soybean oil supplementation would depress adipocyte differentiation. Twenty-eight Angus steers were assigned randomly to 3 groups of 9 or 10 steers and fed a basal diet without additional fat (control), with 3% palm oil (rich in palmitic acid), or with 3% soybean oil (rich in polyunsaturated fatty acids), for 10 wk, top-dressed daily. Palm oil had no effect (P > 0.05) on ADG, food intake, or G:F, whereas soybean oil depressed ADG (P = 0.02), food intake (P = 0.04), and G:F (P = 0.05). Marbling scores tended (P = 0.09) to be greater in palm oil-fed steers (Modest(09)) than in soybean oil-fed steers (Small(55)). Subcutaneous adipocyte mean volume was greater in palm oil-fed steers (515.9 pL) than in soybean-supplemented cattle (395.6 pL; P = 0.01). Similarly, glucose and acetate incorporation into total lipids in vitro was greater in subcutaneous adipose tissue of palm oil-fed steers (119.9 and 242.8 nmol·3h(-1)·10(5) cells, respectively) than adipose tissue of soybean oil-fed steers in (48.9 and 95.8 nmol·3h(-1)·10(5) cells, respectively). Glucose-6-phosphate dehydrogenase and NADP-malate dehydrogenase activities were greater (P ≤ 0.05) in subcutaneous adipose tissue of palm oil-fed steers than in adipose tissue of control steers. Palm oil did not increase palmitic acid or decrease oleic acid in subcutaneous adipose tissue or LM, but decreased (P ≤ 0.05) myristoleic, palmitoleic, and cis-vaccenic acid in adipose tissue, indicating a depression in stearoyl-coenzyme A desaturase activity. Soybean oil increased the proportion of α-linolenic acid in adipose tissue and muscle and increased linoleic acid and 18:1trans-10 in muscle. We conclude that palm oil supplementation promoted lipid synthesis in adipose tissue without depressing feed efficiency or increasing the palmitic acid content of beef.

Winter protein management during late gestation alters range cow and steer progeny performance.

A 4-yr study was conducted at Corona Range and Livestock Research Center, Corona, NM, to establish if a protein-dense self-fed supplement could substitute for a traditional hand-fed (range cube) supplement that is less protein dense and minimize or maintain cow BW and BCS during late gestation and the subsequent steer progeny feedlot performance, health, and economic viability. Late gestation cows received one of 3 supplementation strategies: 1) 36% CP cottonseed meal base supplement (CSM; positive control) fed 3 times per week, 2) self-fed supplement (SMP) comprising 50% animal protein sources (blood meal and feather meal) and 50% trace mineral package, or 3) brief and intermittent supplementation of CSM based on periods of acute environmental stress (VAR; negative control) by ranch management. Initiation of supplementation varied across years due to changing forage conditions and climatically imposed grazing constraints but always ended approximately 2 wks before calving each year. Across all 4 yr, supplement consumption averaged 0.65, 0.21, and 0.04 kg·head(-1)·d(-1) for CSM, SMP, and VAR, respectively. After weaning, steers were preconditioned for 45 d and were received and treated as custom fed commercial cattle at a feedlot in mid November each year. Cow BW and BCS were not influenced (P ≥ 0.13) by prepartum supplementation; however, the strategy did have an effect on BW and BCS change with cows managed in the VAR group. Cows managed in the VAR group lost the greatest (P < 0.05) amount of BW and BCS whereas no differences were measured between CSM and SMP groups. Prepartum supplementation strategies did not influence (P = 0.98) pregnancy rates. Calf weaning, initial feedlot and final BW, and HCW were unaffected (P ≥ 0.80) by prepartum supplementation of the dam. Steers from dams fed CSM and VAR had a greater percentage treated for sickness than SMP steers (P = 0.03), which resulted in a tendency (P = 0.07) for medicine costs to be greater in steers from CSM and VAR cows. The use of a self-fed package supplement was equally effective as use of a traditional hand-fed, oilseed-based supplement in maintaining BW and BCS during late gestation. In addition, these results imply that although nutrition treatment of cows during the prenatal period had no effect on calf growth performance, calves from cows fed SMP had improved feedlot health.

Influence of protein type and level on nitrogen and forage use in cows consuming low-quality forage.

Minimal quantities of ruminally degradable protein from supplements may improve supplement use efficiency of ruminants grazing dormant forages. In Exp. 1, N retention, ruminal NH(3), serum urea N, and NDF digestibility were evaluated for 12 ruminally cannulated cows (Bos spp.) in an incomplete Latin Square design with 3 periods of 42 d each. Cows were fed weeping lovegrass [Eragrostis curvula (Schrad.) Nees] hay (4.1% CP, 75% NDF, OM basis) at 1.3 % BW/d and offered 1 of 3 sources of CP [urea, cottonseed (Gossypium spp.) meal (CSM); or 50% blood meal and 50% feather meal combination (BFM)] fed to supply 0, 40, 80, or 160 g/d of CP. Beginning on d 22 of supplementation, ruminal contents and serum samples were collected at -2, 0, 3, 6, 9, 12, 18, 24, 30, 36, and 48 h relative to the morning offering of hay. On Day 24, feces and urine were collected for 72 h. In Exp. 2, 4 ruminally cannulated steers were used in a replicated 4 by 4 Latin Square to evaluate use of supplements differing in quantity and ruminal CP degradability. Steers were fed 6.8 kg/d chopped sudangrass [Sorghum bicolor (L.) Moench nothosubsp. drummondii (Steud.) de Wet ex Davidse] hay (3.7% CP, 74% NDF on OM basis) and supplemented with 56 g/d of a salt mineral mix (CON); CON + 28 g/d blood meal + 28 g/d feather meal (BFM); CON + 98 g/d CSM (LCS); or CON + 392 g/d CSM (HCS). Treatments provided 0, 40, 40, or 160 g/d of CP for CON, BFM, LCS, and HCS respectively. In Exp. 1, N use and total tract NDF digestibility were not affected by protein sources or amounts (P ≥ 0.18). Ruminal NH(3) concentrations exhibited a quadratic response over time for UREA (P < 0.05) and was greater with increasing inclusion of urea (P < 0.05); whereas BFM or CSM did not differ (P > 0.05) by amount or across time. In Exp. 2, supplementation had a tendency (P = 0.09) to increase DM disappearance. Supplementation also increased (P < 0.01) serum glucose concentrations; however, no difference (P ≥ 0.28) was found between supplements. Serum urea N and ruminal NH(3) concentrations were increased (P ≤ 0.01) in steers fed HCS. Feeding low quantities of a high-RUP supplement maintained rumen function without negatively affecting DM or NDF digestibility of a low-quality forage diet.

Cellular regulation of bovine intramuscular adipose tissue development and composition.

It is well documented that grain feeding stimulates adipogenesis in beef cattle, whereas pasture feeding depresses the development of adipose tissues, including intramuscular (i.m.) adipose tissue. Additionally, production practices that depress adipocyte differentiation also limit the synthesis of MUFA. Marbling scores and MUFA increase in parallel suggesting that stearoyl-coenzyme A desaturase (SCD) gene expression is closely associated with and necessary for marbling adipocyte differentiation. Similarly, marbling scores and fatty acid indices of SCD activity are depressed in response to dietary vitamin A restriction. In bovine preadipocytes, vitamins A and D both decrease glycerol-3-phosphate dehydrogenase (GPDH) activity, an index of adipocyte differentiation, whereas incubation of bovine preadipocytes with l-ascorbic acid-2-phosphate increases GPDH activity. Exposing bovine preadipocytes to zinc also stimulates adipogenesis, putatively by inhibiting nitric oxide (NO) production. However, incubation of bovine preadipocytes with arginine, a biological precursor of NO, strongly promotes differentiation in concert with increased SCD expression. This suggests that the effect of either arginine or zinc on adipogenesis is independent of NO synthesis in bovine preadipocytes. Enhanced expression of SCD is associated with a greater accumulation of MUFA both in bovine preadipocyte cultures and during development in growing steers. In bovine preadipocytes, trans-10, cis-12 CLA strongly depresses adipocyte differentiation and SCD gene expression, thereby reducing MUFA concentrations. The bovine preadipocyte culture studies suggest that any production practice that elevates vitamins A or D or trans-10, cis-12 CLA in bovine adipose tissue will reduce i.m. adipose tissue development. Conversely, supplementation with vitamin C or zinc may promote the development of i.m. adipose tissue.

Effects of supplements that contain increasing amounts of metabolizable protein with or without Ca-propionate salt on postpartum interval and nutrient partitioning in young beef cows.

Cattle grazing winter range forages exhibit interannual variation in response to supplementation. This variation may be mediated by circulating concentrations and subsequent metabolism of glucose, which are influenced by forage quality and availability. A study conducted at the Corona Range and Livestock Research Center during 2 dry years evaluated responses of young postpartum beef cows (n = 51, initial BW = 408 +/- 3 kg, and BCS = 5.1 +/- 0.04 in year 1; n = 36, initial BW = 393 +/- 4 kg, and BCS = 4.5 +/- 0.05 in year 2) to supplements that met or exceeded metabolizable protein (MP) requirements. Supplements were fed at 908 g/d per cow and provided 327 g of CP, 118 g of ruminally undegradable protein (RUP), and 261 g of MP from RUP (RMP), calculated to meet the MP requirement; 327 g of CP, 175 g of RUP, and 292 g of MP from RUP (RMP+), which supplied 31 g of excess MP; or 327 g of CP, 180 g of RUP, 297 g of MP from RUP, and 100 g of propionate salt (NutroCal, Kemin Industries, Inc., Des Moines, IA; (RMP+)P), which supplied 36 g of excess MP. Body weights were recorded once every 2 wk, and blood samples were collected 1x/wk in year 1 and 2x/wk in year 2 for 100 d postpartum. Postpartum anestrous was evaluated by progesterone from weekly blood samples, and pregnancy was confirmed by rectal palpation at weaning. As MP from RUP with or without propionate increased, a decrease (P = 0.03) was observed in postpartum interval; however, differences in pregnancy percentage (P = 0.54) were not influenced by treatments. We hypothesized that additional AA from RUP along with propionate would increase supply of glucogenic precursors and, therefore, glucogenic potential of the diet. Therefore, a postpartum glucose tolerance test was conducted near the nadir of cow BW to evaluate the rate of glucose clearance. Glucose tolerance tests showed that (RMP+)- or (RMP+)P-supplemented cows had greater (P = 0.03) rates of glucose clearance, which might have influenced the observed abbreviation of the postpartum interval. A glucose tolerance test conducted at the end of supplemental treatments revealed no differences in glucose clearance (P = 0.47) among previously supplemented cows. These data suggest that not only vegetative quality, duration of lactation, and season of grazing, but also type of supplementation may play a pivotal role in the young postpartum beef cow's ability to respond and incorporate nutrients into insulin-sensitive tissues.