Brief Summary of Minutes of Annual Meeting

A. Administration: Introductions and administrative details 1. Election of new officers. Promotion of Current Secretary to Chair 2009-2010: Barry Bradford New Secretary 2009-2010: Alex Hristov Next years meeting for NC-1040: Chicago, near OHare, October 25 and 26, 2010. Additional members to invite would be someone from Cornell, Ferguson and Galligan from University of Pennsylvania to strengthen a possible NRC update contribution from this committee. Invitations will be done by Barry Bradford in the coming year. 2. NC Administrative Advisor, David Benfield (also includes summary from Adele Turzillo) The National Institute of Food and Agriculture (NIFA) replaces CSREES and the Agriculture and Food Research Initiative (AFRI) now replaces the National Research Institute (NRI), and essentially combines the NRI and Initiative for Future Agriculture and Food Systems (IFAFS) programs into one. NIFA is being organized into 4 institutes to address 5 primary areas of emphasis. The institutes are food production, environment, food safety, youth and community development. Dr. Benfield suggested people that have not at least turned in a report in the past 3 years should be dropped from the group, because they are not contributing. The final agreed upon protocol is that at least one report is expected from each experiment station. After one year without a report, a warning will be sent by Dr. Benfield to the investigator as well as their experiment station director. After 2 years without a report, Dr. Benfield will be contacted to remove the person from the committee. This will be instituted with initial warnings in 2009, with the first potential removals in 2010. The group decided that there will be no repercussions for lack of attendance, given potential conflicts with schedules that may be a problem every year, although there were mixed feelings on this. The downside is that those who never attend do not likely contribute to the overall goals of coordinating research. The group decided to try the required reports first and re-evaluate in coming years. B. Station Reports 1. Mike VandeHaar (Michigan State University, Obj. 2, 3) An NRC-based formulation spreadsheet (Spartan Dairy 3) was developed and is a stand-alone program. Ease of use of this model and similarity to NRC 2001 suggests that this model would be more useful in the field and in research than the NRC model. Some important differences from NRC are: (1) Requirements are based on body weight adjusted for body condition and gestation; (2) Rumen undegradable protein can contribute to ruminal N by recycling; (3) dietary fat has less negative impact on microbial protein yield; (4) In cases where NRC used a combination of formulas and fixed values for nutrient columns, equations were derived to approximate NRC; (5) fat and carbohydrate fractions can be monitored; and (6) the energy value of protein is lower because Spartan, unlike NRC, assumes most protein is deaminated. The model predicts N and P waste output. An excel file of the model was distributed to all NC-1040 participants. Other work: Studies were completed on intensified calf feeding and productivity/profitability and the role of transforming growth factor- 1in remodeling of mammary tissue during the dry period. Future work: Plan is to better characterize and model depression in digestibility that occurs with increased feed intake, develop better methods to estimate the energy value of protein, and examine effects of specific fatty acids on immune function. Collaborations are planned with OH, WI, VA, and KA. 2. Lou Armentano (University of Wisconsin, Obj. 1) Study 1: Intraruminally infusing branched chain volatile fatty acids had relatively little effect on milk fatty acid composition but isovalerate and anteisovalerate severely depressed feed intake compared to isomolar propionate or acetate. Study 2: Treatments were: Linseed, palm, corn, high-oleic safflower; high-linoleic safflower oil and control. Linoleic acid appeared to be the most potent fatty acid for reducing short chain fatty acid yield. The PALM diet reversed the effect on total fat yield by specifically raising C16 by providing preformed C16. Study 3: Evaluated the effects of corn bran replacing all corn grain in a digestibility study with 24 animals. The digestibility of nutrient fractions (starch, NDF, non-starch NFC) was not greatly affected by diet, except NDF digestibility was decreased in the corn grain, low-forage diet, However, because of increased NDF content in the high corn bran diets, apparent TDN concentration of these diets was lower than those containing corn grain. NRC under-estimated the digestibility of corn grain and over-estimated the digestibility of corn bran. Study 4: Effect of fresh forage composition and protected soy protein supplement to dairy ewes. As the amount of alfalfa in the forage increased the milk, fat, and protein yield increased and the milk urea nitrogen increased. Planned Research: Future work will include measuring blood VFA in Study 1. For Study 2, we have designed, and are conducting, an experiment to specifically test oleic acid vs. linoleic acid that accounts for non-linearity and possible oleic*linoleic interactions. Statistical analyses will be done for Study 4. 3. Gale Batemam (Akey, Obj 1, 3) Study 1: Empirical model development of enteric methane production and comparison to other published models. Current models of methane production are insensitive to dietary effects. Collected 156 observations from 36 trials and put diets into Akeys formulation model. Nutrient densities were fit to published data, and digestibilities were adjusted to make the model correctly predict milk production. Using actual nutrient values is uncommon in most published methane models. Data were evaluated by testing all combinations of predictive models with up to 9 independent variables, then predictive equations were compared and evaluated by using residuals, AIC, and BIC values. Determining the best predictive model without over-parameterization is difficult; AIC can be improved by adding factors, and Hanigan suggested a Chi-square test to evaluate the significance of step-by-step increases in variables included. A variance inflation factor can also be used as well as a traditional step-down multivariate analysis and test significance of individual factors in the total predictive model. The proposed model with 7 factors was considered over-parameterized and many factors had almost no influence on predicted output of methane relative to total production. Discussion also focused on whether beef vs. dairy data needs to be specified in the model, and whether interactions should be evaluated in a greatly reduced model. Planned Research: Future work will include further development of the model in Study 1. 4. Alex Hristov (Pennsylvania State University, Obj. 1) Study 2: The goal of this experiment was to investigate the effect of yeast culture product (Saccharomyces cerevisiae) on rumen fermentation, nutrient utilization, and ammonia and methane emission from manure in dairy cows. Overall, the yeast culture tested had little effect on ruminal fermentation, digestibility, and N losses, but tended to reduce rumen ammonia concentration and increase microbial protein synthesis and decreased ammonia and methane emissions from manure. Study 2: In a continuous culture experiment, glycerin increased propionate and butyrate concentrations and also increased NDF digestibility by up to 10 units (68% vs. 57%). Under present experimental conditions, replacement of starch with dry glycerin seemed to have positive effects, as shown by increased total VFA production, decreased acetate to propionate ratio, and higher DM digestibility. Other Studies: Coconut oil and lauric acid are very effective defaunating agents. However, both also reduce dry matter intake and milk yield, but, due to modified fatty acid profile, may have potential health benefits (i.e. cancer, cholesterol). Decreasing dietary crude protein from 16% to 14% decreased both ammonia emissions from manure and milk yield in dairy cows. Manure from this project was used in an agronomy experiment with growing barley for measurement of ammonia release from manure-amended soil, and even in this situation, the high crude protein diet caused greater ammonia emissions. 5. Jeff Firkins (The Ohio State University, Obj. 1) One hypothesis is that increasing passage rate from the rumen increases outflow of protozoa without promoting so much autolysis. With increasing outflow rate, their growth rate presumably increases to maintain rumen population densities such that they increase efficiency of growth and promote less intraruminal recycling of protozoal and bacterial proteins. If increasing feed intake increases protozoal passage while increasing nutrient supply, as they sense more nutrients, they should up-regulate protein synthesis and cell division. Entodiniomorphs exhibit chemotaxis toward peptides, with no clear differences between various sources of peptides (bacterial, soy, etc.). It is possible binding of nutrients (cellular receptors) activates an intracellular signaling cascade which results in release of NO, subsequent activation of cGMP and altered cilial beating (i.e. movement). Actual engulfment of particles using fluorescent beads is being currently investigated. The hypothesis is that PI3K activation during chemotaxis is involved in moving digestive vacuoles toward cytoproct via intracellular calcium and calmodulin gradients altering cytoskeletal trafficking. In the micronucleus, protozoa have approximately 5 chromosomes of which one chromosome contains genes encoding ribosomal synthesis and function. Multiple copies of these chromosomes are copied into numerous fragments in the macronucleus, with ribosomal DNA having thousands of copies per cell. The relative abundance of rDNA chromosomal fragments with different diets / nutrients is being studied. Planned Research: 1. Study protozoal chemotaxis to integrate with growth response in a working model (collaborating with J. Knapp). Specifically evaluating Ca++-mediated signaling and integrating chemotaxis with uptake of inert, COO--charged beads (about the size of bacteria and with a similar charge). 2. Use pulsed-field electrophoresis and cDNA libraries to characterize the genomes of two prominent rumen ciliates (with S. Karnati and Z. Yu). 3. Determine efficiency of bacterial amino acid incorporation/biosynthesis in mixed continuous cultures (working with B. Bequette). 4. Development of a model to integrate protozoal predation of bacteria, protozoal growth rate, and protozoal lysis rates (collaborating with B. Bequette and plan to consult with M. Hanigan and J. Knapp). 6. Arnold Hippen (South Dakota University, Obj. 1 and 2) Study 1: The hypothesis for this research was feeding a source of unsaturated fat, distillers grains (DC), in combination with highly fermentable starch, high moisture corn (WC), will create a ruminal environment leading to incomplete biohydrogenation of fatty acids and milk fat depression. The experiment was a 2x2 factorial assessing effect of fat from distillers grains versus ruminally inert fat and fermentability of starch from high moisture corn versus dry corn. Diets including 20% alfalfa hay and 30% corn silage. There were no effects on DMI. Distillers grains decreased acetate to propionate ratio in the rumen, but did not did alter milk yield. Milk fat concentrations were decreased with DG. The WC decreased milk yield, and WC and DG interacted to cause a dramatic decrease in milk fat concentrations. The elsewhere reported relationship of trans-10 C18:1 to milk fat depression was observed and there were indications of the relationship of trans-10, cis-12 C18:2 to milk fat depression, but statistical significance was not obtained. Research Planned: Ruminal degradability of protein and lipid in oilseeds processed with alternative methods of heat treatment in dairy cow diets. Effects on lactation performance. 7. Shawn Donkin (Purdue, Obj. 1 and 2) Study 1 (Obj. 1): Mixtures of 25, 50, or 75% WDG were evaluated for pH, stability, and acetate concentration in 30 day re-ensiled haylage or corn silage. WDG increased stability of haylage, but decreased stability of corn silage. Study 2 (Obj. 2): When propionate is added to hepatocytes, portions of the PEPCK promoter region become protected from DNase I digestion, suggesting binding of transcription factors. Study 3 (Obj. 2): Heat stressed bovine hepatocytes have increased expression of PC, consistent with responses in vivo. Bovine PC is expressed as 6 transcriptional variants, and luciferase reporter constructs for each were transfected into rat hepatoma cells. Promoter 3 appeared to increase (not significant), while promoter 1 significantly decreased during heat stress in these cells; net mRNA abundance of rat PC is decreased in heat-stressed rat hepatoma cells. This response does not appear to be consistent between the in vitro models. Study 4 (Obj. 2): Identification of the propionate response sequence for bovine PEPCK. The genomic sequence of the bovine PEPCK promoter was cloned by PCR and the resulting sequence analyzed. Collectively these data demonstrate potential regulation of hepatic propionate metabolism by induction of specific transcription factor binding sites in the PEPCK promoter. Planned Research: Work related to objective 1 and 2 will continue by continuing and extending the research outlined above. 8. J. W. Schroeder (North Dakota State University, Obj. 1) The objective was to determine the effect of processing method on digestibility, ruminal fermentation, in situ degradation of alfalfa hay and rate of passage of flaxseed in diets of Holstein steers fed a lactating dairy cow diet. Four ruminal and duodenal cannulated Holstein steers were used in a 4 x 4 Latin square design to determine the effect of processing on ruminal fermentation and digestibility of nutrients in dairy cattle fed whole, rolled, or ground flax. Steers were fed conventional corn and forage diets that contained 50% concentrate (DM basis). Treatments included: 1) no flaxseed, 2) whole flaxseed, 3) rolled flaxseed, and 4) ground flaxseed. The control diet contained linseed meal at proportions equal to the amount of protein contributed by flaxseed, and chromic oxide at 0.25% of the DM to determine digesta flows at the duodenum. Feed, ruminal, duodenal, and fecal samples were collected for determination of effects on ruminal fermentation parameters including ammonia, pH, and volatile fatty acids, as well as ruminal and total tract digestibilitys of dry matter, crude protein, NDF, ADF, and fat. Planned Research: (1) To determine if the direct dietary supplementation of dry field peas can replace protein and energy supplements in the diets of calves (CSREES/CSFL). (2) Replace soybean meal with ethanol byproducts and locally grown pulse crops in the diets of lactating dairy cattle (NDSFC). 9. Marshall Stern (University of Minnesota, Obj. 1) Study 1: Effects of two halophytic plants (Kochia and Atriplex) on fermentation by ruminal microbes in vitro were examined. Results showed no differences in dry matter and crude protein digestibility between the plants. Atriplex had higher organic matter (OM) digestibility compared with Kochia. Neutral detergent fiber digestibility of Atriplex (41.1%) was higher than Kochia (34.8%), however acid detergent fiber digestibility was higher in Kochia (40.6 vs 23.4%). There were no differences between plants in molar proportion of acetate and propionate, but butyrate and valerate in Kochia were two fold of Atriplex. Protein synthesis was higher with Kochia compared with Atriplex (5.96 vs 4.85 g of N/kg of OM digested). Study 2: Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in cows and continuous culture were evaluated. Yeast tended to decrease total ruminal VFA (mM) and increased ruminal pH (6.3 vs 6.5). In continuous culture, yeast decreased ammonia N concentration and flow, but had no other effects. Beneficial effects of Saccharomyces cerevisiae on rumen pH and NH3-N concentration appear to be dependent upon diet type. Planned Research: 1. To quantify ruminal and post-ruminal distribution of protected trace minerals (Cu, Zn, Mn) in purified diets with or without lignosulfonate. 2 To determine the interaction between dietary sulfur and dietary roughage on microbial fermentation. 10. Brian Bequette (University of Maryland, Obj. 2) Study 1: Lactating dairy cows were used to determine whether dietary choline (Reashur@) spares Met by increasing Met re-methylation. [1-13C] and [13CH3] Met tracers were infused to determine Met de- and re-methylation. Based on plasma [1-13C]homocysteine as precursor pool, methyl group flux was 2-fold higher than carboxyl group flux. Dietary choline reduced methyl flux, thus reducing Met catabolism. Study 2. We showed that GIT tissues of sheep possess enzymes of the urea cycle and that ammonia detoxification to urea is activated in these isolated cells by N-carbamoylglutamate (NCG). This study aimed to demonstrate that NCG improves urea-N capture in ruminants. Wether sheep were given continuous intravenous infusions of either saline or NCG for 10 d. Plasma arginine and urea were increased by NCG, while gluconeogenesis from non-glucose sources was reduced (25%) by NCG. Planned Research: We plan to determine if de novo fatty acid (FA) synthesis is the primary factor limiting milk triglyceride (TG) synthesis. Two experiments will test this hypothesis under CLA induced down-regulation of mammary TG synthesis. The 1st study in lactating mice will use Rosiglitazone, an inducer of lipid synthetic pathways, to rescue milk fat during CLA induced milk fat depression. A 2nd study in lactating cows will test whether post-ruminal infusion of short and medium chain FA can rescue milk fat TG synthesis when CLA is given. 11. Barry Bradford (Kansas State University, Obj. 1 and 2) Study 1: Continued work investigating the potential for molasses to help prevent milk fat depression. Molasses replaced corn grain at 5% of diet DM in a very low-fiber / high-starch diet. Molasses increased milk fat content but effects on milk fat yield were not significant. Molasses decreased yield of trans-10 C18:1 and increased yield of trans-11 C18:1, consistent with increased use of the normal biohydrogenation pathway. Molasses decreased total ruminal VFA concentration and increased ruminal pH. b>Study 2: The effects of postruminal niacin infusion were evaluated in 6 growing steers. Feed intake dropped dramatically over 4 days of infusion. Tissues were analyzed for abundance of GPR109a, the niacin receptor. Unlike in humans and rodents, the receptor was found in liver and muscle tissue in addition to adipose tissue. In a separate study, the effects of encapsulated niacn (EN) were tested in 22 transition cows. EN was top-dressed at 24 g/day from 21 days before calving through 21 days in milk. EN decreased postpartum NEFA and BHBA, but also decreased feed intake of multiparous cows in the week prior to calving. Depression of intake by niacin may be related to the broad distribution of its receptor in the bovine. Planned Research: 1. Evaluate effects of VFA infusions on expression of ANGPTL4 by ruminal epithelial cells. 2. Determine whether TNF alpha decreases hepatic glucose production in lactating cows. 12. Mark Hanigan (Virginia Polytechnic Institute & State University, Obj. 2 & 3) Study 1 (Obj. 2). The objective of this study was to investigate the effects of essential amino acids (EAA) and acetate on the phosphorylation status (PS) of mammalian target of rapamycine (mTOR), and ribosomal protein S6 (rpS6) in MAC-T cells. Extracellular EAA availability appeared to have a stronger effect on protein synthesis signaling in MAC-T cells than extracellular acetate. Study2 (Obj. 2). The objective of this study was to investigate the total and individual effects of essential amino acid (EAA) on protein synthesis signaling inMAC-T cells. Essential amino acid deprivation, particularly Arg, Phe, Trp, and branched chain amino acids significantly reduced signaling for translation initiation via mTOR and rpS6 in MAC-T cells. Study 3 (Obj. 3). Cell Signalling Model: The objective of this work was to develop a mathematical representation of mTOR dependent insulin and EAA signaling. This mathematical model included six protein pools representing phosphorylated and unphosphorylated forms of Akt, mTOR, and 4EBP1. The model was built in AcslXtreme. Mass action equations were used to represent phosphorylation (FUP,P) and dephosphorylation (FP,UP) fluxes. Other model development: Derivation of rate parameters for ruminal digestion of nutrients and microbial growth have been derived from the data set used to parameterize the digestive elements of the 2001 NRC model. Planned research: 1. Evaluate mammary signaling pathway responses to methionine, lysine, leucine, and combinations of these amino acids in vivo. 2. Complete the cell signaling model and incorporate it into an existing mammary tissue model with the objective of testing whether it improves predictions of milk protein synthesis. 13. Heidi Rossow for Jim Fadel (UCD, Obj. 3) Study 1: Improve Molly energy prediction by updating ATP constants in some pathways. Global sensitivity analysis assists in understanding the effects of changes in model and effects on basal metabolic rate predictions. Some stoichiometries were updated. The adjusted model predicts slight increase in daily milk yield compared to previous model, but effect on blood FA is dramatic, and indicates instability due to lack of adequate energy. Body fat is especially sensitive to model estimate of ATP yield from glucose. Study 2: Measure ammonia emissions from manure over 24 hours and correlate results with MUN from animal or herd. Specific gravity and dietary CP can be used to predict urine urea N (UUN) with reasonable accuracy, but at very high dietary CP, relationship between specific gravity and UUN is different than in lower CP diets. Data from 1521% CP diets showed that higher CP diets led to higher manure pH, greater ammonia release, and lower proportions of ammonia N to total N in urine. Study 3: Develop N kinetics in manure slurries, incorporating composition of manure, time effects, and storage volume effects. Data from in vitro will be used to parameterize this model. Planned Research: Collaboration with Hanigan, McNamara, and Johnson in global sensitivity analyses, model development, and better maintenance partitioning using new ATP yields, work on in vitro data collection of NH3 with modeling, and develop an optimization model for diary. C. Station Report Summaries for members not at the meeting 1. Donald Beitz (ISU, Obj. 2). Feeding viable-NP51®, not HK-NP51®, increases production of anti-inflammatory cytokine IL-10 in non-infected mice only. This attribute has significant implications on treatment of autoimmune diseases. Interestingly, when administered to mice challenged with MAP, NP51® behaves differently as illustrated by decrease of the MAP-specific IL-10 production (anti-inflammatory) and increase of the MAP-specific IFN-³ (pro-inflammatory) (Figures 1 and 2). Noteworthy, for tissue dissemination of MAP, high concentrations of IL-10 and low concentrations of IFN-³ are required. Planned Research: Will examine the performance of periparturient dairy cows fed the NP51 during early and mid lactation periods. Milk quality, udder health and immune competence will be closely evaluated during the NP51-feeding time. 2. Keith Cummins (Auburn University, Obj. 2). This year gels were run continuing proteomics evaluation of muscle tissue with the onset of lactation. The gels run provided the following protein sets: Set 1: Acidic to neutral proteins and housekeeping cytosolic proteins. Most soluble proteins such as cytosolic p. Set 2: Intermediate solubility proteins. Some overlap with set 1. Set 3: Membrane proteins. Generally insoluble in reagents to obtain first two sets. These are generally highly expressed proteins. The gels have been run. Waiting on protein identification from colleagues at UAB and proteomics lab that does sequencing and identification via MALDI-TOF. Visually gels are very different and various subsets were run to identify any proteins that were potentially covered up by actin and myosin of our previous gels. Planned research: Intend to sequence and identify exposed proteins and will do statistical analysis of effects of lactation on the proteins identified. Then the research will be broadened to include the effects of lactation onset on hepatic proteins and will be done using serial biopsies of livers of mature lactating dairy cows at -14, +7 and + 28 days in milk. 3. Erdman, Richard (University of Maryland) See report under B (10). 4. Eun, Jong-Su (Utah State University, Obj. 1). Study 1: To test magnesium zeolite as source of ruminal buffer additive, thirty lactating Holstein cows were assigned equally to one of 3 dietary treatments: control diet (TMR), TMR diet with 1.4% sodium bicarbonate (SBD), and TMR diet with 1.4% zeolite (ZLD). DM intake and milk yield did not differ across treatments. Ruminal pH increased with ZLD compared to the control and was similar to cows fed SBD. Economically, zeolite could replace sodium bicarbonate as a ruminal buffer in lactating dairy diet. Study 2: Assess effect of replacing alfalfa hay (AH) with birdsfoot trefoil (BFT) on fermentation in vitro with three dietary treatments: 100% AH; 50% AH +50% BFT (AHBFT); and 100% BFT. Methane production was not influenced by treatments. NH3-N concentration decreased in cultures offered AHBFT compared with AH, and decreased in cultures receiving BFT compared with AH and AHBFT. Total VFA was not affected by diet. Feeding BFT altered the metabolic pathways of in vitro ruminal fermentation with beneficial modification of N use and cultures fed BFT did not have any negative impact on ruminal fermentation. Study 3: Objectives were to determine if supplementing commercial tannic acid (CT) extract affected N utilization and CH4 production, and to assess if CT supplementation would prevent in vitro ruminal acidosis when mixed cultures offered a barley-based high concentrate diet. Methane production increased in response to increasing CT, whereas NH3-N concentration decreased. Total VFA increased with increasing CT. Supplementation of CT resulted in accelerated microbial production. Planned Research: Continued research with CT. Lactational performance of dairy cows fed brown mid-rib corn silage-based diet or conventional corn silage and nonforage fiber sources diets. 5. James Fadel (UC, Davis) See report under B (13). 6. Gabriella Varga (Pennsylvania State University) See report under B (4). 7. John McNamara, (Washington State University, Obj. 2 and 3). Primary inadequacies in Molly are in description of subtle metabolic pattern differences in the most efficient animals. Work has improved: 1) interaction of genetic merit and dietary energy on efficiency of dairy cattle; 2) transcriptomic analysis of adipose tissue as it relates to control of energy efficiency, 3) use of metabolic modifiers to improve efficiency and 4) improvement of the model to describe control of reproductive efficiency by nutrients. The collaborator is on sabbatical leave to work on the model and the gene transcription array analyses, cooperation with from CA, MD and VA. Planned Research: 1) to improve inadequacies in estimation of the energetic cost of visceral and muscle protein turnover, ion transport and the energetic cost of increased feed intake (and rapidly changing feed intake. 2) Integration transcriptional control of energy use. A data set of 48 gene transcriptions arrays in adipose tissue from 21 d prepartum to 56 d postpartum will be used. 3) Integration of reproductive processes to extend the overall description of efficiency from just nutrient use to reproductive efficiency There will be collaboration with members at CA, MD and VA on these works 8. Donato Romagnolo, (The University of Arizona, Obj. 2) This study investigated the mechanisms of transcriptional activation of COX-2 by the AhR and the reversal effects of dietary aromatic hydrocarbon receptor (AhR) antagonists. Small-interfering RNA (siRNA) for the AhR prevented TCDD-induced binding of the AhR to the COX-2 and CYP1A1 promoter regions. The binding of the AhR to COX-2 oligonucleotides from the COX-2 promoter was repressed by cotreatment with the synthetic AhR antagonists, 3-methoxy-4-naphthoflavone (3M4NF), and dietary agents DIM and resveratrol. Chromatin immunoprecipitation (ChIP) assays revealed that the treatment with TCDD induced the time-dependent association of the AhR, the histone acetylase p300, acetylated histone-H4, and phosphorylated H3-Ser10 with the COX-2 promoter. In contrast, in ChIP assays we observed that the cotreatment with DIM antagonized the recruitment of the AhR and phosphorylated H3-Ser10 to the COX-2 promoter. Planned Research: The association of the AhR to xenobiotic responsive elements (XRE) harbored in the BRCA-1 promoter region contributed to repression of BRCA-1 transcription and reduced BRCA-1 protein levels. These changes were associated with loss of histone-4 acetylation, increased recruitment of histone deacetylase-1 and methyl-binding proteins. Dietary antagonists of the AhR with antagonistic effects comprise the phytoalexin resveratrol and 3,3-diindolylmethane (DIM). Resveratrol reduced AhR recruitment in a dose-dependent fashion and restored BRCA-1 expression. Chromatin modifications induced by the AhR at the BRCA-1 gene were reversed by resveratrol. These studies suggest that AhR antagonists such as resveratrol may be useful in the prevention of epigenetic silencing induced by AhR agonists in mammary epithelial cells.

D. NC 1040 5-year Accomplishments and Impact: The need as indicated by stakeholders. Over 55% of the calcium, 17% of the protein, and 15% of the energy in the US diet are supplied by dairy products; thus, the US consumer is a major stakeholder for the NC-1040 committee. Consumers want dairy products that are safe and inexpensive, but increasingly they also want an environmentally friendly dairy industry that promotes animal well-being. Recently, attention has been given to bioactive molecules in milk (in addition to Calcium) such as conjugated linoleic acids (CLA). Yet at the core the NC-1040 committee functions to do basic and applied research on the feeding and nutritional biology of dairy cattle. Major stakeholders include other scientists, practicing nutritionists, veterinarians, and farmers. The needs of these stakeholders have been addressed by the Food Animal Integrated Research group in the FAIR 2002 document. The goals of FAIR 2002 are to strengthen global competitiveness, enhance human nutrition, protect animal health, improve food safety and public health, ensure environmental quality, and promote animal well-being. Because feed inputs are a major determinant of milk yield, cow health, feed efficiency, profitability, and waste output, the work of the NC-1040 committee is critical for most of these goals. The concentration of dairy animals into larger units is an established and continuing trend. This concentration makes some waste management issues more prominent but also more manageable. The importance of our work. Natural resources are used efficiently when milk production per unit feed and per cow is high. To efficiently produce milk, a cow must have a well-developed mammary gland and be able to supply the gland with the nutrients it needs. Nutrition in the first year of life affects mammary gland development, and nutrition around the time of calving and throughout lactation has a major effect on the health, productivity, and efficiency of cows. Feeding for optimal nutrient intake requires not only the provision of the necessary nutrients for milk production but also consideration to the effects of diet on mammary capacity and on appetite, health, and metabolic regulation of the cow. Because feed costs account for half of all costs on a dairy farm, nutrition also significantly impacts farm expenses. The NC-1040 committee considers all of these factors for optimal feeding. For example, if we could maintain current milk production while feeding diets with 4 percentage units less total protein, we would decrease N losses to the environment in the US by 470,000 metric tons per year and save US dairy farmers $1 billion per year in feed costs. This type of progress only can be made if we take an integrated approach, with the use of mechanistic bio-mathematical models that accurately describe metabolism and production of cows. Integration of results. This committee has a proven track record of making significant impacts in our knowledge dairy cattle nutrition and metabolism and in the way that dairy cattle are fed and managed nationwide. We use the same approach that has proven effective in the past: that is to challenge and refine our models of dairy nutrition and metabolism. Computer-based, mechanistic, and quantitative metabolic models are useful in two ways: first, they help us determine critical needs in research and second they enable practical improvements in dairy cow feeding. Critical research needs are determined by using existing data from NC-1040 members or conducting new experiments to test model predictions of physiological responses to experimental diets. Examples of such responses include, rumen pH, microbial growth and function; alterations in gene expression and hormonal release of organs such as the adipose tissue; and alterations in milk fatty acid compositions. By challenging our working models in this way, we identify shortcomings that then become the basis for developing new testable hypotheses for further experimentation. Results from new experiments are incorporated into the models, and they are challenged again for further refinement. Thus, we continue to build our models so they are more mechanistic, quantitative, and accurate. These qualities enable us to improve practical feeding recommendations for dairy cattle in a variety of environmental and feeding conditions. Need for Cooperative Work. Important and complex problems require coordinated effort of many personnel. Considerable progress has been made in dairy nutrition, but practical problems remain and no single research group has the skills and resources needed to solve them alone. Only through cooperation can State Experiment Stations address the complex interactions among feed supply, nutrient use, genetic capability, and milk composition. Our committee is comprised of dairy scientists with a broad base of specialties that encompass feed analysis, feeding management, ruminal microbial metabolism, intestinal digestion, physiology and metabolism of splanchnic, adipose, muscle, and mammary tissues, endocrine regulation, molecular and cellular biology, and mathematical modeling. Furthermore, in testing and refining nutrition models for the whole country, we must consider the variation in forages and environment that exist among regions. Thus, we have scientists from every dairy region in the country. In addition, the explosion of new information in genomics, gene expression, gene array work, metabolomics and proteomics requires that we integrate this knowledge into our understanding of metabolic efficiency. Cooperation among stations is required to deal with this information and to solve problems, and will have a national impact in understanding the complex interrelationships of nutrient digestion and metabolism in lactating dairy cows and to apply this knowledge. Impacts on Science and Other Impacts. This project exemplifies the proven effectiveness of the cooperative regional approach. As detailed in the "Related Current and Previous Work" section below, results of this cooperative effort have become benchmarks of scientific progress and have led to practical feeding recommendations used worldwide. Project Leaders for the NC-1040 regional project have received numerous awards for research, both basic and practical, from the American Dairy Science Association, the American Society of Animal Sciences, and industry groups. Most of the Project Leaders are in continuous demand as speakers for scientific and industry conferences in nutrition. The impact on basic and practical nutrition from Project Leaders has been profound in the areas of starch and protein chemistry and nutrition, feed processing, nutrient metabolism, and lactation biology. This group provided a major contribution to the 2001 version of the National Research Councils (NRC's) Nutrient Requirements of Dairy Cattle. Four of the 10 scientists on the NRC panel were from the NC-1040 committee (IL, MD, NH, PA), and a significant portion of the data used in the latest edition came from NC-1040 committee members. In 2005, the group presented a symposium at ADSA/FASS on regulation of nutrient use in dairy cattle. Thus, this committee has had a major impact on improving the biological, economical, and environmental efficiency of the US dairy industry. E. Summary of Progress: Funds Leveraged to Support Work on Project in 2009. The research conducted year-on-year of this project could not be possible without significant funding support from various avenues. Members of the committee have a long history of acquiring competitive funds from federal and state agencies, and indeed, not a year has gone by over the last 20 years that at least one member of the project has not held a USDA-NRI grant. In 2009, members of the committee leveraged a total of $1,987,096 from various agencies and private industry to support research activities. This total breaks down into the categories of: Federal funds: $1,142,168 State funds: $99,632 College funds: $16,375 International agencies: $9,000 Boards/Councils/Association funds: $127,000 Private Industry funds: $592,921 Research Activities and Progress. One overriding goal in feeding cattle is to find the optimal combination of chemical and physical properties of feeds that provides the proper amount and balance of absorbed nutrients to match the ability given by the genotype of the cow or herd. This is a major challenge because of the tremendous variety of feedstuffs available, their complexity of interactions among feed particles, nutrients and organisms in the rumen, genetic variance within and among herds, and the rapidly changing nutrient requirements of a cow around the time of parturition. The amount and profile of absorbed nutrients in dairy cattle are a function of rumen bacterial fermentation and intestinal digestion. Feed particles and microbes that escape the rumen can be digested in the small intestine to produce amino acids, monosaccharides, and lipids for absorption. The chemical and physical properties of feeds determine the rumen bacterial and protozoal populations and the end-products they produce, and, relatedly, the availability of nutrients critical to the production of milk and milk components in a variety of ways. For example, the chemical composition (including total protein, nonprotein nitrogen, amino acid balance, organic acids, lipids, fiber, and non-fiber carbohydrate) dictates directly the availability of nutrients to support rumen microbial growth and the absorbed nutrients available to the animal to support milk synthesis. The physical properties of feeds, either inherent in the plant structure or altered by various processing methods, alters degradability in the rumen, and thus determines the proportion of feed fermented and used for rumen microbial growth and the proportion that passes to the small intestine. The cow is a fully integrated metabolic system in which one, even minor, change in nutrient input may lead to a variety of downstream events that alters function at the tissue and whole animal levels. Since the last revision, we now understand more fully that this also includes changes in gene expression, and endocrine responses that were unknown or just discovered 5 years ago (IGF-1, leptin, perilipin, cytokines and ghrelin from the adipose tissue, for example). (KS, MI, WA) Dietary carbohydrate fractions differ in the profiles of glucogenic and lipogenic metabolites they yield upon rumen bacterial metabolism and intestinal absorption. The amount and types of carbohydrates also impact rumen pH, which, in turn, alters fermentation and can alter the yield of nutrients, even amino acids, for absorption. Thus the various carbohydrate fractions have differential effects on the yield and composition of milk. Recent improvements in methods will allow more accurate prediction of optimal amounts and ruminal availability of non-fiber carbohydrates for efficient production of milk and milk components. More importantly, because of the integrative modeling approach of this committee, we have a quantitative knowledge of the maximal percentage contribution of these fractions to overall yield and efficiency on different diets, and can move on to further work. The amount and balance of absorbed amino acids also helps determine milk yield, not just milk protein synthesis. This availability of amino acids, in turn, is a function of the amount of feed protein which passes undegraded through the rumen and the amount of ruminally synthesized microbial protein that reaches the small intestine. Because microbial protein has a better amino acid profile than many feed proteins, this remains an important area of study. Microbial protein yield is also a function of the amount and type of organic matter fermented. Thus, microbial protein yield varies by source of carbohydrate and protein, and rate of fermentation. This is a classic example of the need for an integrated approach to dairy cattle nutrition-we must continue to design experiments across state lines that allow a full scope of study of the key variables. We need to continue to build a comprehensive model that explicitly includes these types of interactions. Synthesis of milk and milk components is a function of both the synthetic potential of the mammary gland and the supply of metabolites to the mammary gland. Supply of metabolites comes from dietary components, some of which are modified in other tissues, and from mobilization of body lipids and amino acids. There is an interaction between metabolism of body tissues, the supply of dietary nutrients and the milk production potential of the cow (as well as other animals) which has been recognized for quite some time which provide an extreme range of response of animals to even the same diet. While many dairy scientists have been slow to recognize the importance of these interactions, several stations in this project have been studying these interactions across a range of diets, genetic potentials, and stages of lactation (AL, CA, IA, IN, KS, MI, PA,WA and more recently OH, MD, VA). Data has been used to refine our feeding recommendations on a wide variety of feedstuffs. New concepts on the interactions of nutrition and gene expression is exemplified with work from several stations: At IN new information on molecular control of enzymes that modulate hepatic gluconeogenesis reveal specific differences between the cow and other animals. At WA, work with supplemental chromium, a nutrient known to be required for many years, a positive feed intake and milk production response was obtained with supplemental chromium, along with a reduction in lipolysis and an increase in lipogenesis in adipose tissue, removing the negative effects of fatty acid mobilization on feed intake in early lactation. Many nutritionists have now recognized that we cannot do relevant nutritional research without integrating this work with genetics and gene expression. Many nutrients are now known to affect gene expression in several organs, which then alters the animal response to the diet or further changes in the diet. Newer additions to the committee (KS) as well as adapting previous members (AL, IA, IN, MD, MI, OH, VA, WA) have begun serious efforts in identifying genetic responses to diet and to lactation. This work falls presently into the basic aspect-providing hard data to other scientists and advanced professionals on the key interactions of genetics and diet. This holds future promise in even more efficient feeding management and breeding programs. Major advancements have occurred in our knowledge of the interaction of metabolism and the endocrine system. Studies at AL, IN, and MI, in collaboration with other NC-1040 members, have illustrated the role of nutrition in the IGF-I system of dairy cattle. At IA, the role of glucagon in lipid metabolism has shown its potential for treatment for fatty liver while recent work at KS has begun to provide a mechanistic basis for the involvement of adipose derived cytokines in initiation of fatty liver. Work has also been done on the role of leptin in mammary gland development and regulation of feed intake (MI). If we are to improve the accuracy and precision of predicting nutrient use, we must continue to improve mechanistic, dynamic models of metabolism. The newer Dairy Nutrient Requirements book (NRC, 2001) was based in large part on data from this committee. In the 7 years since its publication, it has gained great respect in the industry. However, the process of revision of this document, and the model within it, also pointed out many of the shortcomings in our current knowledge base. The new version is limited especially in predicting dietary nutrient interactions, which consequently hamper our ability to predict rumen microbial metabolism and microbial protein yield and therefore responses to rumen-undegraded protein, carbohydrate, and fat supplements. Other significant limitations are the ability to predict short term versus long-term nutritional responses and changes in body fat and protein use. More mechanistic modeling of the metabolism of the lactating dairy cow will allow for evaluation of these interactions. The most comprehensive mechanistic and dynamic model of metabolism in the dairy cow is called 'Molly', developed at CA with inputs from most NC-1040 members. Members of this project (IL, MD, NH, PA) also have been instrumental in developing the new NRC model (NRC, 2001), which serves as the standard for dairy ration formulation and evaluation in the US. These different computer nutrition programs are currently in use for predicting nutrient requirements and productivity of lactating dairy cows. While all of these systems are soundly based on available data, all have weaknesses in the areas defined by Objectives 1 and 2. The rate of degradation of feedstuffs, the effect of various dietary carbohydrates on rumen fermentation and microbial protein synthesis, and quantitative data on metabolic interchanges among nutrients and body tissues limit the accuracy of these systems. New collaborative efforts by the NC-1040 project are needed to remove these inaccuracies. Molly is limited in its ability to describe the rapid changes in nutrient use that occur in early lactation and in predicting physiological responses to high feed intakes or diets with atypical amino acid, fiber, or starch contents. This is not surprising, given the paucity of these types of data when the model was originally constructed in the 1970 and 1980s. The modeling work done spurred new research into getting those data. Work done by several members of the committee (VA, MD, OH) is being used to challenge Molly for its description of energy use in the viscera and mammary gland. Visceral metabolism can account for the majority of maintenance requirements and can be highly variable. Errors in the model reflected a lack of knowledge of visceral metabolism in early and mid lactation. Using data generated, challenges and improvements to the model describing energy use have been made, further increasing its utility in research and application (McNamara, 2005, 2006). Quantitative data are still needed on the supply of milk component precursors available under different metabolic and nutritional conditions, such as early lactation. Data also are needed on the metabolic interconversions of nutrients, such as the use of amino acids for gluconeogenesis and thus milk lactose synthesis (MD, VA), and the partitioning of body fat and fat derived from the diet or lipogenesis for milk fat synthesis (WA). These data will enable further refinement of current nutrition recommendations and aid in interpretation of feeding experiments. F. Other Specific accomplishments: Reductions in feeding levels for ruminally degradable protein would reduce nitrogen losses in manure and improve animal efficiency (VA). As ammonia emissions from manure are driven by the amount of urinary N deposited in manure, such changes would lead to reduced ammonia emissions from animal and manure storage facilities. Improved knowledge of the mechanisms that regulate milk protein synthesis will allow development of models that better predict the requirements for milk protein synthesis. This in turn will allow more refined estimates of N requirements and reduced safety margins in feeding systems. Such an outcome will also work to reduce ammonia emissions from animal and manures storage facilities. Phosphorus availability in the digestive tract is an important determinant of the amount that must be fed and the amount that is lost in feces. Model development has helped identify key aspects of P digestion that warrant further examination and must be considered in requirement models to achieve greater reductions in P feeding levels (VA). This research evaluates the relationships between milk urea nitrogen, plasma urea nitrogen and urine urea nitrogen. Milk urea N can be a used to predict UUN excretion and may be extended to estimate NH3 emissions from dairy cattle manure because there is a strong relationship between UUN excretion and NH3 emissions. The information from this research is being used to test metabolic models for urine urea excretion (CA). Greater use of flax in the nutrition of dairy cattle can supplement lactation diets with not only protein and energy, but compliment the growing interest in designer foods with milk enriched with omega-3 and omega-6 from such grains as flax seed. Furthermore, preliminary evidence suggests that dairy cow fed flax also have improved reproductive health with improved pregnancy rates. It has been estimated that if dairy cow pregnancy rates could be increased, an estimated cost savings to the dairy enterprise of $8.73 per cow per year could be realized for every percentage unit gained (ND).

H. Publications of NC-1040 Committee members during 2009 reporting year (does not include papers in press or abstracts) Project Collaborative Refereed Publications: Bobe, G., A. R. Hippen, P. She, G. L. Lindberg, J. W. Young, D. C. Beitz. 2009. Effects of glucagon infusions on protein and amino acid composition of milk from dairy cows. J. Dairy Sci. 92:130-138. Bobe, G., J. C. Velez, D. C. Beitz, and S. S. Donkin. 2008. Glucagon Increases Hepatic mRNA Levels of Gluconeogenic and Ureagenic Enzymes in Early Lactation Dairy Cows. J. Dairy Sci. 92:5092-5099 Cyriac, J., A. G. Rius, M. L. McGilliard, R. E. Pearson, B. J. Bequette, and M. D. Hanigan. 2008. Lactation performance of mid-lactation dairy cows fed ruminally degradable protein at concentrations lower than NRC recommendations. J. Dairy Sci. 91:4704-4713. (Impact Factor: 2.361). Hanigan, M. D., J. France, S. J. Mabjeesh, W. C. McNabb, and B. J. Bequette. 2009. High rates of mammary tissue protein turnover in lactating dairy goats are energetically costly. J. Nutr. 139:1118-1127. Individual Station Refereed Publications: Abdelqader, M. M., A. R. Hippen, K. F. Kalscheur, D. J. Schingoethe, K. Karges, and M. L.Gibson. 2009. Evaluation of corn germ from ethanol production as an alternative fat source in dairy cow diets. J. Dairy Sci. 92:1023-1037. Allen, M. S., B. J. Bradford, and M. Oba. 2009. BOARD-INVITED REVIEW: The hepatic oxidation theory of the control of feed intake and its application to ruminants. J Anim Sci. 87(10):3317-34. Araujo, R.C., A.V. Pires, I. Susin, C.Q. Mendes, G.H. Rodrigues, I.U. Packer, and M.L. Eastridge. 2008. Milk yield, milk composition, eating behavior, and lamb performance of ewes fed diets containing soybean hulls replacing coastcross (Cynodon species) hay. J. Anim. Sci. 86:3511-3521. Autken, S.K., Karcher, E.L., Rezamand, P., Gandy, J.C., VandeHaar, M.J., Capuco, A.V., and Sordillo, L.M. 2009. Evaluation of antioxidant and proinflammatory gene expression in bovine mammary tissue during the periparturient period. J Dairy Sci 92: 589-598. Bach, A., M. Ruiz Moreno, M. Thrune and M. D. Stern. 2008. Evaluation of fermentation dynamics of soluble crude protein from three protein sources in continuous culture fermenters. J. Anim. Sci. 86:1364-1371. Baik, M., B.E. Etchebarne, J. Bong and M. J. VandeHaar. 2009 Gene Expression Profiling of Liver and Mammary Tissues of Lactating Dairy Cows. Asian-Aust. J. Anim. Sci. 22: 871-881. Bateman, II, H. G., T. M. Hill, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Effects of corn processing, particle size, and diet form on performance of calves in bedded pens. J. Dairy Sci. 92:782-789. Bionaz, M., C. R. Baumrucker, E. Shirk, J. P. Vanden Heuvel, E. Block, and G. A. Varga. 2008. Characterization of mardi-darby bovine kidney cell line for ppars: temporal response and sensitivity to fatty acids. J. Dairy Sci. J. Dairy Sci. 91:2802-2813. Bobe, G., G. L. Lindberg, L. F. Reutzel, and M. D. Hanigan. 2009. Effects of lipid supplementation on the yield and composition of milk from cows with different ²-lactoglobulin phenotypes. J. Dairy Sci. 92:197-203. (Impact Factor: 2.361). Bradford, B. J., L. K. Mamedova, J. E. Minton, J. S. Drouillard, and B. J. Johnson. 2009. Daily injection of tumor necrosis factor alpha increases hepatic triglycerides and alters transcript abundance of metabolic genes in lactating dairy cattle. J Nutr. 139(8):1451-1456. Broderick, G. A., N. D. Luchini, S. M. Reynal, G. A. Varga, and V. A. Ishler. 2008. Effect on production of replacing dietary starch with sucrose in lactating dairy cows. J. Dairy Sci. 91:4801-4810. Chang E, S.S. Donkin, and D. Teegarden. 2009. Parathyroid hormone suppresses insulin signaling in adipocytes. Mol Cell Endocrinol. 307:77-82. Chaves, A. V. C., Mao Long M.L. He, W. Z. Yang, A. N. Hristov, T. McAllister, and C. Benchaar. 2008. Effects of essential oils on proteolytic, deaminative and methanogenic activities of mixed ruminal bacteria. Can. J. Anim. Sci. 88:117-122. Chung, Y.-H., Pickett, M.M., T. W. Cassidy, and G. A. Varga. 2008. Effects of prepartum dietary carbohydrate source and monensin on periparturient metabolism and lactation in multiparous cows. J. Dairy Sci. J. Dairy Sci. 91:2744-2758. Clark, J. H., R. A. Christensen, H. G. Bateman, II and K. R. Cummings. 2009. Effects of sodium sesquicarbonate on dry matter intake and production of milk and milk components by Holstein cows. J. Dairy Sci. 92:3354-3363. Degner SC, Papoutsis AJ, Selmin O, Romagnolo DF. Targeting of aryl hydrocarbon receptor-mediated activation of cyclooxygenase-2 expression by the indole-3-carbinol metabolite 3,3'-diindolylmethane in breast cancer cells. J Nutr. 2009 Jan; 139(1):26-32. Dekking, L., F. Koning, D. Hosek, T. D. Ondrak, S. L. Taylor, J. W. Schroeder, and M. L. Bauer. 2009. Intolerance of celiac disease patients to bovine milk is not due to the presence of T cell stimulatory epitopes of gluten. Nutr. 25:22-23. DeVuyst E. A., M. L. Bauer, F. C. Cheng, J. Mitchell, D. Larson. 2008. The impact of a leptin gene SNP on beef calf weaning weights. Anim Genet. 39:284-286. Diaz, H.L., A.M. Stalford, and J.L. Firkins. 2009. Differential chemotaxis by entodiniomorphids and isotrichids toward peptides of bacterial, protozoal, and soy origin. Microb. Ecol. 57:567-568. Donkin, S. S., S. Koser, H. White, P. H. Doane, and M. J. Cecava. 2009. Feeding value of glycerol as a replacement for corn grain in rations fed to lactating dairy cows. J. Dairy Sci. 92:5111-5119. Eastridge, M.L., P.B. Bucci, and C.V.D.M. Ribeiro. 2008. Feeding equivalent concentrations of forage neutral detergent fiber from alfalfa hay, grass hay, wheat straw, and whole cottonseed in corn silage based diets to lactating cows. Anim. Feed Sci. Technol. 150:86-94. El-Kadi, S.W., Baldwin. R.L. VI, McLeod, K.R., Sunny, N.E., and Bequette, B.J. (2009) Glutamate is the major anaplerotic substrate in the tricarboxylic acid cycle of isolated rumen epithelial and duodenal mucosal cells from beef cattle. J. Nutr. 139: 869-875. Fae, G.S., R. M. Sulc, D.J. Barker, R.P. Dick, M.L. Eastridge, and N. Lorenz. 2009. Integrating winter annual forages into a no-till corn silage system. Agron. J. 101 (5):1286-1296. Hanigan, M. D., C. C. Palliser, and P. Gregorini. 2009. Altering the representation of hormones and adding consideration of gestational metabolism in a metabolic cow model reduced prediction errors. J. Dairy Sci. 92:5043-5056. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2008. Crude Protein for Diets Fed to Weaned Dairy Calves. Prof. Anim. Sci. 24: 596-603. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Effects of Consistency of Nutrient Intake from Milk or Milk Replacer on Dairy Calf Performance. Prof. Anim. Sci. 25: 85-92. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Effects of Changing the Essential and Functional Fatty Acid Intake of Dairy Calves. J. Dairy Sci. 92:670-676. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Roughage for diets fed to weaned dairy calves. Prof. Anim. Sci. 25: 283-288. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Optimizing nutrient ratios in milk replacers for calves less than five weeks of age. J. Dairy Sci. 92:3281-3291. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Effects of fat concentration in a high-protein milk replacer on calf performance. J. Dairy Sci. 92:5147-5153. Hill, T. M., H. G. Bateman, II, J. M. Aldrich, and R. L. Schlotterbeck. 2009. Effect of weaning age of dairy calves fed a conventional or more optimum milk replacer program. Prof. Anim. Sci. 25: 619-624. Horn NL, Donkin SS, Applegate TJ, Adeola O. 2009. Intestinal mucin dynamics: response of broiler chicks and White Pekin ducklings to dietary threonine. Poult Sci. 88:1906-1914. Hristov, A.N., C.E. Basel, A. Melgar, A.E. Foley, J.K. Ropp, C.W. Hunt, and J.M. Tricarico. 2008. Effect of exogenous polysaccharide-degrading enzyme preparations on ruminal fermentation and total tract digestibility of nutrients in lactating dairy cows. Anim. Feed Sci. Technol. 145:182-193. Hristov, A.N., J.K. Ropp, S. Zaman, and A. Melgar. 2008. Effect of essential oils on ruminal fermentation and ammonia release in vitro. Anim. Feed Sci. Technol. 144:55-64. Kadegowda A.K.G., M. Bionaz, L.S. Piperova, R. A. Erdman, and J. J. Loor. 2009 Peroxisome proliferator-activated receptor-gamma activation and long-chain fatty acids alter lipogenic gene networks in bovine mammary epithelial cells to various extents J. Dairy Sci. 92: 4276-4289. Kadegowda. A. K. G., M. Bionaz, B. Thering, L. S. Piperova, R. A. Erdman, and J.J. Loor. 2009. Identification of Internal Controls for Quantitative PCR in Mammary Tissue of Lactating Cows Receiving Lipid Supplements J. Dairy Sci. 92:2007-2019. Karnati, S.K.R., J.T. Sylvester, C.V.D.M. Ribeiro, L.E. Gilligan, and J.L. Firkins. 2009. Investigating unsaturated fat, monensin, or bromoethanesulfonate in continuous cultures retaining ruminal protozoa. I. Fermentation, biohydrogenation, and microbial protein synthesis. J. Dairy Sci. 92:3849-3860. Karnati, S.K.R., Z. Yu, and J.L. Firkins. 2009. Investigating unsaturated fat, monensin, or bromoethanesulfonate in continuous cultures retaining ruminal protozoa. II. Interaction of treatment and presence of protozoa on prokaryotic communities. J. Dairy Sci. 92:3861-3873. Kozelov, L.K., F. Iliev, A.N. Hristov, S. Zaman, and T.A. McAllister. 2008. Effect of fibrolytic enzymes and an inoculant on in vitro degradability and gas production of low-dry matter alfalfa silage. J. Sci. Food Agric. 88:2568-2575. Lardy, G. P., B. A. Loken, V. L. Anderson, D. M. Larson, K. R. Maddock-Carlin, B. R. Ilse, R. Maddock, J. L. Leupp, R. Clark, J. A. Paterson, and M. L. Bauer. 2009. Effects of increasing field pea (Pisum sativum) level in high-concentrate diets on growth performance and carcass traits in finishing steers and heifers. J Anim Sci. 87:3335-3341. McPhee, M. J., J. W. Oltjen, J. G. Fadel, D. G. Mayer and R. D. Sainz. 2009. Parameter estimation and sensitivity analysis of fat deposition models in beef steers using acslXtreme. Mathematics and Computers in Simulation. 79:2701-2712. Mikolayunas-Sandrock, C., L. E. Armentano, D. L. Thomas, and Y. M. Berger. 2009. Effect of protein degradability on milk production of dairy ewes. J Dairy Sci 92: 4507-4513. Mullins, C. R., K. N. Grigsby, and B. J. Bradford. 2009. Effects of alfalfa inclusion rate on productivity of lactating dairy cattle fed wet corn gluten feed based diets. J Dairy Sci. 92(7):3510-3516. Mulrooney, C. N., D. J. Schingoethe, K. F. Kalscheur, A. R. Hippen. 2009. Canola meal replacing dried distillers grains with solubles in lactating dairy cow diets. J. Dairy Sci. 92:5669-5676. Ndegwa, P.M., A.N. Hristov, J. Arogo, and R.E. Sheffield. 2008. A Review of Ammonia Emissions Mitigation Techniques for Concentrated Animal Feeding Operations. Biosystems Engineering, 100:453  469. Oelker, E.R., C. Reveneau, and J.L. Firkins. 2009. Interaction of molasses and monensin in alfalfa hay- or corn silage-based diets on rumen fermentation, total tract digestibility and milk production by Holstein cows. J. Dairy Sci. 92:270-285. Oliver, C. E., B. K. Magelky, M. L. Bauer, F. C. Cheng, J. S. Caton, H. Hakk, G. L. Larsen, R. C. Anderson, D. J. Smith. 2008. Fate of chlorate present in cattle wastes and its impact on Salmonella Typhimurium and Escherichia coli O157:H7. J Agric Food Chem. 56:6573-6583. Riasi, A., M. Danesh Mesgaran, M. Ruiz Moreno, M. D. Stern. 2008. Chemical composition, in situ ruminal degradability and post-ruminal disappearance of dry matter and crude protein from the halophytic plants Kochia scoparia, Atriplex dimorphostegia, Suaeda arcuata and Gamanthus gamacarpus Anim. Feed Sci. Technol. Vol. 141/3-4:209-219. Siddiqui SM, Chang E, Li J, Burlage C, Zou M, Buhman KK, Koser S, Donkin SS, Teegarden D. 2009. Dietary intervention with vitamin D, calcium, and whey protein reduced fat mass and increased lean mass in rats. J Steroid Biochem Mol Biol. 112:122-126. Singh, K., R. A. Erdman, K. M. Swanson, A. J. Molenaar, N. J. Maqbool, T. T., J. Arias, E. Quinn-Walsh, and K. Stelwagen. 2009. Epigenetic Regulation of Milk Production in Dairy Cows. J. Mammary Gland Biol. Neoplasia. 14 (In Press). Sylvester, J.T., S.K.R. Karnati, B.A. Dehority, M. Morrison, G.L. Smith, N.R. St-Pierre, and J.L. Firkins. 2009.Rumen protozoa decrease generation time and adjust 18S rDNA copies to adapt to decreased transfer interval, starvation, and monensin. J. Dairy Sci. 92:256-269. Tabe E. S., J. Oloya, D. K. Doetkott, M. L. Bauer, P. S. Gibbs, M. L. Khaitsa. 2008. Comparative effect of direct-fed microbials on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle. J Food Prot. 71:539-544. Taylor, M. S., K. F. Knowlton, M. L. McGilliard, W. S. Swecker, J. D. Ferguson, Z. Wu, and M. D. Hanigan. 2009. Dietary calcium has little effect on mineral balance and bone mineral metabolism through 20 weeks of lactation in Holstein cows. J. Dairy Sci. 92:223-237. (Impact Factor: 2.361). Teegarden D and S. S. Donkin. 2009. Vitamin D: emerging new roles in insulin sensitivity. Nutr Res Rev. 22:82-92. Teegarden D., and S.S. Donkin. 2009. Vitamin D: emerging new roles in insulin sensitivity. Nutr Res Rev. 22:82-92. Thorson, J. F., B. J. Karren, M. L. Bauer, C. A. Cavinder, J. A. Coverdale, and C. J. Hammer. 2009. Effect of Selenium Supplementation and Dietary Energy Manipulation on Mares and Their Foals: Foaling Data. J. Anim. Sci. (Accepted) Thrune, M., A. Bach, M. Ruiz-Moreno, M.D. Stern and J.G. Linn. 2009. Effects of saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows. Livestock Sci. Vol. 124:261-265. Toshniwal, J. K., C. D. Dechow, B. G. Cassell, J. A. D. R. N. Appuhamy, and G. A. Varga. 2008. Heritability of electronically recorded daily body weight and correlations with yield, dry matter intake and body condition score. J Dairy Sci. 91:3201-3210. Vander Pol, M., A. N. Hristov, S. Zaman, N. Delano, C. Schneider. 2008. Effect of inclusion of peas in dairy cow diets on ruminal fermentation, digestibility, and nitrogen losses. Anim. Feed Sci. Technol. 150:95105. Vyas, D., and R. A. Erdman. 2009. Meta-analysis of milk protein yield responses to lysine and methionine supplementation. J Dairy Sci. 92:5011-5018. Zeng, R., Bequette, B.J., Vinyard, B.T., and Bannerman, D.D. (2009) Determination of milk and blood concentrations of lipopolysaccharide-binding protein in cows with naturally acquired subclinical and clinical mastitis. J. Dairy Sci. 92: 980-989. Conference Proceedings, Theses, and Popular Press Articles: Abdelqader, M. 2008. Evaluation of Feeding Fat from Corn Coproducts on Performance of Lactating Dairy Cows. Ph.D. Dissertation, South Dakota State University, Brookings. 132 pp. Appuhamy, J.A.D.R.N., A.L. Bell, J. Escobar, and M. D. Hanigan. 2009. Effects of essential amino acid deprivation on protein synthesis signaling in bovine mammary epithelial cells in-vitro. Pp424-425 in XIth International Symposium on Ruminant Physiology (Chilliard, Y. Glasser, F. Faulconnier, Y., Bocquier, F., Veissier, I., and Doreau, M., eds.). Wageningen Academic Publishers, Wageningen Gale Bateman. 2009. Plot your course, but know where the exits are located. Progressive Dairyman. Issue 9, June 12, 2009. pp 46-47 Garcia, A. D., A. R. Hippen. 2008. Alimentación de las vacas lecheras para condición corporal. SDSU Extension Extra, EXEX4040s, June. Garcia, A. D., A. R. Hippen. 2008. Feeding dairy cows for body condition score. SDSU Extension Extra. ExEx. 4040, June. Garcia, A. D., A. R. Hippen. 2008. Preventative feeding of the dairy cow in transition Dairy Star,12/27, pg. 16 Garcia, A. D., A. R. Hippen. 2009. Alimentación preventiva de la vaca lechera en transición. Albeitar. No. 125. 10-12. Garcia, A. D., A. R. Hippen. 2009. Preventative feeding of the dairy cow in transition. Progressive Dairyman, Issue 4. Pp. 13-15. Garcia, A. D., K. F. Kalscheur, A. R. Hippen, and K. Rosentrater. 2009. O problema resultante da presença de micotoxinas em grãos de destilaría destinados a rumiantes. Albeitar: publicacion veterinaria independendiente, Portugal, March/April, No. 2. pp. 48-53. Garcia, A. D., K. F. Kalscheur, A. R. Hippen, D. J. Schingoethe, K. Rosentrater. 2008. Mycotoxins in corn distillers grains: a concern in ruminants? SDSU Extension Extra, ExEx4038, March Garcia, A. D., K. F. Kalscheur, A. R. Hippen, K. Rosentrater. 2008. Micotoxinas en granos de destileria. Una preocupacion en rumiantes? SDSU Extension Extra, ExEx4038-S, April Garcia, A. D., K. F. Kalscheur, A. R. Hippen, R. Schafer. 2008. High priced corn and dairy cow rations. Progressive Dairyman. 11:1-3. Schingoethe, D. J., A. D. Garcia, K. F. Kalscheur, A. R. Hippen, and K. Rosentrater. 2009. El azufre en los granos de destilería para el ganado lechero. South Dakota State University, Cooperative Extension Service. ExEx4039S. Schroeder, J.W. 2009. Cold Temps Hinder Silage Production. Hay & Forage Grower. October 29. http://hayandforage.com/silage/corn/1029-cold-silage-production/index.htm. Schroeder, J.W. 2009 Cold Temps Hinder Silage Production. Hay & Forage Grower. Dairy Today magazine and E-moo Dairy Newsletter. October 12. Schroeder, J.W. 2009. Preservatives make wet hay useable. September 14. Feedstuffs Vol 81, No 40, p 25 Schroeder, J.W. 2009. Organic acids help protect corn. December 14. Feedstuffs Vol 81, No 51, p 11-12. Schroeder, J.W. 2009. Surviving in a down economy. MaxYield Cooperative. February 10. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d201%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM Schroeder, J.W. 2009. Dont let pre-weaned dairy calves go hungry. MaxYield Cooperative. February 17. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d209%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Proper harvesting, storage impacts silage quality. MaxYield Cooperative. July 7. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d403%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Watch for signs of depression. MaxYield Cooperative. August 4. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d441%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Heifer synchronization programs can be cost effective. MaxYield Cooperative. August 11. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d446%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM Schroeder, J.W. 2009. Dairy farm managers dare to compare. MaxYield Cooperative. September 8. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d481%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Stop feed loss to birds. MaxYield Cooperative. October 6. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d520%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Stop feed loss to birds. MaxYield Cooperative. Dairy Today. September 21. http://www.dairyherd.com/ForageCN.asp?contentid=837536 Schroeder, J.W. 2009. Preservatives make wet hay useable. MaxYield Cooperative. October 6. http://www.maxyieldcooperative.com/maxyield/news.aspx?MaxYield=WebPointID%3d0%26NewsStoryID%3d514%26__TimeStamp__%3d12%2f31%2f9999+11%3a59%3a59+PM. Schroeder, J.W. 2009. Feed Frozen 'Silage' Carefully. Hay & Forage Grower and eCornSilage Newsletter. November 24. http://hayandforage.com/ecorn-archive/1124-frozen-silage-scientist-warns/ index.html Sumner, J. M., C. Schachtschneider, A. Hutjens, A. Youngquist, G.Duncan, S. Rocco, S., J. Miller, J. L Vierck and J.P. McNamara. 2009. Regulation of dairy cattle adipose tissue metabolism by adrenergic control systems and gene transcription mechanisms dictating increased overall efficiency. International Society of Ruminant Digestion and Physiology, Clermont-Ferrand, France, September 2009. Schactschneider, C., Youngquist, A., Rocco, S. M., Vierck, J. L. and J. P. McNamara. 2009. Using a dynamic metabolic model to investigate patterns of nutrient flux in the most efficient dairy animals and to integrate gene expression data into metabolic control. 7th International Workshop on Modeling Nutrient use in Farm Animals, Paris, France, Williams, C. M., C. M. Dschaak, J.-S. Eun, A. J. Young, and J. W. MacAdam. 2009. Ruminal metabolism during continuous culture fermentation when replacing alfalfa (Medicago sativa L.) hay with birdsfoot trefoil (Lotus corniculatus L.) hay. Pages 148-151 in Proceedings, Western Section, American Society of Animal Science, Colorado State University, Fort Collins, CO.