R. C. Waterman

Next Generation Sequencing to Define Prokaryotic and Fungal Diversity in the Bovine Rumen

A combination of Sanger and 454 sequences of small subunit rRNA loci were used to interrogate microbial diversity in the bovine rumen of 12 cows consuming a forage diet. Observed bacterial species richness, based on the V1–V3 region of the 16S rRNA gene, was between 1,903 to 2,432 species-level operational taxonomic units (OTUs) when 5,520 reads were sampled per animal. Eighty percent of species-level OTUs were dominated by members of the order Clostridiales, Bacteroidales, Erysipelotrichales and unclassified TM7. Abundance of Prevotella species varied widely among the 12 animals. Archaeal species richness, also based on 16S rRNA, was between 8 and 13 OTUs, representing 5 genera. The majority of archaeal OTUs (84%) found in this study were previously observed in public databases with only two new OTUs discovered. Observed rumen fungal species richness, based on the 18S rRNA gene, was between 21 and 40 OTUs with 98.4–99.9% of OTUs represented by more than one read, using Good’s coverage. Examination of the fungal community identified numerous novel groups. Prevotella and Tannerella were overrepresented in the liquid fraction of the rumen while Butyrivibrio and Blautia were significantly overrepresented in the solid fraction of the rumen. No statistical difference was observed between the liquid and solid fractions in biodiversity of archaea and fungi. The survey of microbial communities and analysis of cross-domain correlations suggested there is a far greater extent of microbial diversity in the bovine rumen than previously appreciated, and that next generation sequencing technologies promise to reveal novel species, interactions and pathways that can be studied further in order to better understand how rumen microbial community structure and function affects ruminant feed efficiency, biofuel production, and environmental impact.

Reproductive performance of heifers offered ad libitum or restricted access to feed for a one hundred forty-day period after weaning.

Reproductive performance was evaluated in composite heifers born over a 3-yr period that were randomly assigned to control (fed to appetite; n = 205) or restricted (fed at 80% of that consumed by controls adjusted to a common BW basis; n = 192) feeding for a 140-d period, beginning about 2 mo after weaning at 6 mo of age and ending at about 12.5 mo of age. Heifers were fed a diet of 67% corn silage, 18% alfalfa, and 9% of a protein-mineral supplement (DM basis). Restricted heifers consumed 27% less feed over the 140 d and had less ADG (0.53 +/- 0.01 vs. 0.65 +/- 0.01 kg/d; P < 0.001) than control heifers. After 140 d, all heifers were placed in common pens and subjected to an estrous synchronization protocol to facilitate AI at about 14 mo of age. Heifers were then exposed to bulls for the remainder of a 51-d breeding season. Average BW of heifers diverged within 28-d after initiation of feed restriction, and differences (P < 0.001) persisted through the prebreeding period (309 +/- 1 vs. 326 +/- 1 kg at approximately 13.5 mo of age) and subsequent grazing season (410 +/- 2 vs. 418 +/- 2 kg at about 19.5 mo of age). From the end of the 140-d restriction at about 12.5 to 19.5 mo of age, ADG was greater (P < 0.001) in restricted heifers than control heifers (0.51 +/- 0.01 vs. 0.47 +/- 0.01 kg/d). Proportion of heifers attaining puberty by 14 mo of age tended to be less (P = 0.1) in restricted (60 +/- 3%) than control-fed heifers (68 +/- 3%). Mean BW at puberty was less (P < 0.01) in restricted (309 kg) than control (327 kg) heifers. Pregnancy rate from AI tended to be less (P = 0.08) in restricted (48 +/- 4%) than control heifers (57 +/- 3%). Proportion of animals that were pubertal at breeding and pregnant from AI were positively associated (P < 0.1) with heifer age and ADG from birth to beginning of study. Final pregnancy rates were 87 and 91% for restricted and control heifers, respectively (P = 0.27). Day of breeding season that conception occurred was negatively associated with ADG from birth to weaning (P = 0.005), but was not associated with ADG within treatment (P = 0.60). Economic analysis revealed a

Preovulatory estradiol and the establishment and maintenance of pregnancy in suckled beef cows.

33 reduction in cost to produce a pregnant heifer under the restricted protocol when accounting for pregnancy rates and differences in BW and market prices between selection at weaning and marketing as open heifers at l.5 yr of age. A potential economic advantage exists for rearing replacement heifers on a restricted level of feeding during the postweaning period.

Effects of flunixin meglumine on pregnancy establishment in beef cattle.

In postpartum beef cows, GnRH-induced ovulation of small dominant follicles decreased pregnancy rates and increased late embryonic/fetal mortality. In Exp. 1, single ovulation reciprocal embryo transfer (ET) was used to examine the relationship between preovulatory serum concentrations of estradiol at GnRH-induced ovulation in donor and recipient cows and establishment and maintenance of pregnancy. Suckled beef cows (n = 1,164) were administered GnRH (GnRH1, 100 μg) on d -9 (GnRH1), PGF(2α) on d -2, and GnRH2 (GnRH2, 100 μg) on d 0 (CO-Synch protocol) either with (donors; n = 810) or without (recipients; n = 354) AI. Single embryos (n = 394) or oocytes (n = 45) were recovered from the donor cows (d 7; ET) and all live embryos were transferred into recipients. Serum concentration of estradiol at GnRH2 was positively correlated with follicle size at GnRH2 (r = 0.45, P < 0.01) and progesterone at ET (r =0.34, P < 0.01). Donor cows with greater estradiol at GnRH2 were more likely to yield an embryo than an unfertilized oocyte (P < 0.01). Donor and recipient cows were retrospectively divided into 4 groups [low estradiol (<8.4 pg/mL) or high estradiol (≥8.4 pg/mL)] based on serum concentration of estradiol at GnRH2. Pregnancy rate at d 27 for low-low (n = 78), low-high (n = 80), high-low (n = 91), and high-high (n = 101) groups (donor-recipient, respectively) was 45, 65, 43, and 61% respectively (P < 0.02). Because recipient cows with greater estradiol concentration at GnRH2 had greater pregnancy rates in Exp. 1, the objective of Exp. 2 was to evaluate the effect of estradiol supplementation on pregnancy rate. Ovulation was synchronized in suckled beef cows (n = 600) using the CO-Synch protocol with the insertion of a controlled internal drug release (CIDR; intravaginal progesterone supplement) from d -9 until d -2. Approximately one-half of the cows (n = 297) received an injection of estradiol cypionate (ECP; 0.5 mg intramuscularly) 24 h before AI. Compared with the no treatment (Control) cows, ECP treatment increased (P < 0.01) pregnancy rates of cows induced to ovulate smaller dominant follicles (<12.2 mm). In conclusion, GnRH-induced ovulation of small dominant follicles was associated with reduced serum estradiol, fertilization rate (donor cows), and pregnancy establishment (recipient cows). Furthermore, ECP supplementation during the preovulatory period increased pregnancy rates in cows induced to ovulate smaller dominant follicles.

Relationship between body condition score at calving and reproductive performance in young postpartum cows grazing native range

The objective of this research was to determine effects of a single injection of the PG synthesis inhibitor flunixin meglumine (FM; 1.1 mg/kg of BW, intramuscularly) approximately 13 d (range 10 to 15 d) after AI on pregnancy establishment. Three experiments were conducted using estrus-synchronized heifers and cows. Technicians and AI sires were equally represented across treatments within locations and experiments. Bulls were introduced on the day of FM treatment. Pregnancy to AI was diagnosed 28 to 50 d after AI using ultrasonography. In Exp. 1, beef heifers (n = 1,221) were divided within 5 locations to receive FM or no further treatment (control). At insemination, heifers were divided into 2 similar pastures or pens, and approximately 13 d later, 1 group of heifers within each location was processed through an animal handling facility to administer FM treatment. There was no location x treatment interaction (P = 0.62) on AI pregnancy rates, so data were pooled. Pregnancy rates to AI were reduced (P = 0.02) among heifers receiving the FM treatment procedure (66%) compared with control heifers (72%). In Exp. 2, suckled beef cows (n = 719) were assigned within 2 locations to receive FM or no further treatment (control) approximately 13 d after AI. At insemination, control and FM cows were divided into separate pastures, and only FM cows were handled after AI for the FM treatment procedure. There was no location x treatment interaction (P = 0.75), so data were pooled. Pregnancy rates to AI did not differ (P = 0.80) between FM (57%) and control cows (59%). In Exp 3, beef heifers (n = 247) and suckled beef cows (n = 335) from 1 location received no injection (control) or injection of FM approximately 13 d after AI when all cows and heifers were processed through a working facility. Pregnancy rates to AI were not different (P = 0.37) between FM (45%) and control (42%) cows or between FM (56%) and control (55%) heifers. We conclude FM administration at 1.1 mg/kg of BW approximately 13 d after AI did not improve pregnancy establishment in beef cows and heifers and that the effects of handling heifers at this time may decrease pregnancy establishment.

Protein and glucogenic precursor supplementation: A nutritional strategy to increase reproductive and economic output

Body condition score is used as a management tool to predict competency of reproduction in beef cows. Therefore, a retrospective study was performed to evaluate association of BCS at calving with subsequent pregnancy rate, days to first postpartum ovulation, nutrient status (assessed by blood metabolites), and calf BW change in 2- and 3-yr-old cows (n = 351) managed and selected to fit their environment of grazing native range over 6 yr at the Corona Range and Livestock Research Center, Corona, NM. Cows were managed similarly before calving, without manipulation of management, to achieve predetermined BCS at parturition. Palpable BCS (scale of 1 to 9) were determined by 2 experienced technicians before calving. Cows were classified to 1 of 3 BCS groups prior calving: BCS 4 (mean BCS = 4.3 ± 0.02), 5 (mean BCS = 5.0 ± 0.03), or 6 (mean BCS = 5.8 ± 0.06). Cows were weighed weekly after calving and serum was collected once weekly (1 yr) or twice weekly (5 yr) for progesterone analysis to estimate first postpartum ovulation beginning 35 d postpartum. Year effects also were evaluated, with years identified as either above or below average precipitation. Days to first postpartum ovulation did not differ among calving BCS groups (P = 0.93). Pregnancy rates were not influenced by calving BCS (P = 0.83; 92%, 91%, 90% for BCS 4, 5, and 6, respectively). Days to BW nadir was not influenced by BCS at calving (P = 0.95). Cow BW was different at all measuring points (P < 0.01) with BCS 6 cows having the heaviest BW and cows with BCS 4 the lightest. Cows with calving BCS 4 and 5 lost more (P = 0.06) BW from the initiation of the study to the end of breeding than cows with BCS 6. However, cow BW change at all other measurement periods was not different (P ≥ 0.49) among calving BCS groups. Serum glucose and NEFA concentrations were not influenced by calving BCS (P ≥ 0.51). Calf BW at birth (P = 0.60), branding (55-d BW; P = 0.76), and weaning (205-d BW; P = 0.60) were not impacted by cow calving BCS. Body condition score did not influence overall pregnancy rates, indicating that young cows can have a reduced BCS and still be reproductively punctual. Therefore, these results indicate that reproductive performance of young cows with reduced BCS may not be affected when managed in extensive range conditions.

Beef Species Symposium: potential limitations of NRC in predicting energetic requirements of beef females within western U.S. grazing systems.

Reproductive performance in young beef cows is often compromised due to a mismatch of physiological demands and suboptimal environmental conditions. Studies conducted at the Corona Range and Livestock Research Center from 2000 to 2007 evaluated 3 postpartum supplement strategies that varied in the amount of glucogenic potential (GP) supplied. Reproductive variables, milk production, and serum metabolites were used to assess supplement effectiveness and economics associated with 2- and 3-yr-old beef cows (n = 379) grazing native range. Supplements were individually fed twice/week at 1,135 g/d (2003 to 2004) or 908 g/d (all other years) and provided 1) 327 g of CP, 109 to 118 g of RUP (CON); 2) 327 to 341 g of CP, 142 to 157 g of RUP (RUP); or 3) 327 g of CP, 151 to 173 g of RUP + 40 to 100 g of propionate salt (PS; RUP+PS). Ultimately, total GP for CON, RUP, and RUP+PS was 44 to 47, 57 to 70, and 93 to 141 g, respectively. Blood samples were collected once/week (2000) or twice/week (2001 to 2007) for progesterone analysis to estimate days to resumption of estrus. Cows were exposed to bulls for 60 d or less, and pregnancy was confirmed by rectal palpation at weaning. Days to resumption of estrus after calving decreased linearly (P = 0.02), resulting in an increased pregnancy rate (P = 0.03) with increasing GP. Milk production exhibited a quadratic (P = 0.04) response to increasing GP, with cows fed RUP producing the most amount of milk. However, a linear decrease (P = 0.07) in days from BW nadir to estrus was found with increasing GP. Total kilograms of calf weaned per cow exposed for the supplemental year and subsequent year was increased linearly (P = 0.07) with increased GP. The improvement in pregnancy rate by supplementing RUP+PS resulted in an increase in total revenue of 18% compared with CON-fed cows and 9.5% compared with RUP-fed cows in the subsequent year after supplementation. These data suggest feeding young cows additional GP in the form of PS allows for partitioning of nutrients away from milk production and toward reproduction, allowing for increased profitability by increasing pregnancy rates and decreasing days to resumption of estrus.

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

Assessment of beef cow energy balance and efficiency in grazing-extensive rangelands has occurred on a nominal basis over short time intervals and has not accounted for the complexity of metabolic and digestive responses; behavioral adaptations to climatic, terrain, and vegetation variables; and documentation of the effects of nutrient form and supply to grazing cattle. Previous research using pen-fed cows demonstrated differences (P < 0.01) in efficiency of weight change ranging from 135 to 58 g/Mcal ME intake. Furthermore, variation in efficiency of ME use for tissue energy gain or loss ranged from 36% to 80%. In general, energy costs for maintenance, tissue accretion, and mobilization were greatest in Angus-based cows, intermediate in Brahman- and Hereford-based cows, and least in dairy-based cows. The most efficient cattle may reflect the types that are successful in semiarid grazing environments with low input management. Successful range cattle systems are likely the result of retention of animals that best adapted to the grazing environment and thus were potentially more efficient. Animals exposed to a variety of stressors may continually adapt, so energy expenditure is reduced and may tend to depart from the modeled beef cow in the 1996 NRC Beef Cattle Requirements. Critical factors comprising cow lifetime achievement, including reproductive success, disease resistance, and calf weaning weight, may be driven by cow total energy utilization in energy-limiting environments. Therefore, energy adjustments for adapted cattle within these landscapes and seasonal BW changes can alter seasonal NEm requirements. Evaluated studies indicate that in static grazing environments, NRC prediction fitness was improved compared with predictions from dynamic systems where cattle were influenced less by management and more by environmental conditions. Preliminary herd analyses cast doubt on the accuracy of NRC BCS descriptions representing NEm requirements of adapted females utilizing semiarid rangelands. Possible gaps are proposed that could be the basis for prediction inaccuracies. A more complete understanding of mechanisms contributing to productivity in the field than the current model predicts will improve future models to better simulate energetic accountability and subsequent female performance.

BEEF SPECIES SYMPOSIUM: An assessment of the 1996 Beef NRC: Metabolizable protein supply and demand and effectiveness of model performance prediction of beef females within extensive grazing systems123

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.

Sources of variability in livestock water quality over 5 years in the Northern Great Plains

Interannual variation of forage quantity and quality driven by precipitation events influence beef livestock production systems within the Southern and Northern Plains and Pacific West, which combined represent 60% (approximately 17.5 million) of the total beef cows in the United States. The beef cattle requirements published by the NRC are an important tool and excellent resource for both professionals and producers to use when implementing feeding practices and nutritional programs within the various production systems. The objectives of this paper include evaluation of the 1996 Beef NRC model in terms of effectiveness in predicting extensive range beef cow performance within arid and semiarid environments using available data sets, identifying model inefficiencies that could be refined to improve the precision of predicting protein supply and demand for range beef cows, and last, providing recommendations for future areas of research. An important addition to the current Beef NRC model would be to allow users to provide region-specific forage characteristics and the ability to describe supplement composition, amount, and delivery frequency. Beef NRC models would then need to be modified to account for the N recycling that occurs throughout a supplementation interval and the impact that this would have on microbial efficiency and microbial protein supply. The Beef NRC should also consider the role of ruminal and postruminal supply and demand of specific limiting AA. Additional considerations should include the partitioning effects of nitrogenous compounds under different physiological production stages (e.g., lactation, pregnancy, and periods of BW loss). The intent of information provided is to aid revision of the Beef NRC by providing supporting material for changes and identifying gaps in existing scientific literature where future research is needed to enhance the predictive precision and application of the Beef NRC models.