METABOLIC RELATIONSHIPS IN SUPPLY OF NUTRIENTS FOR LACTATING COWS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1017181
• Multistate No.: NC-_old2040
• Project Start Date: Oct 1, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Project Director
Hristov, AL, NI.

Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802

Performing Department
Animal Science

Non Technical Summary
The US dairy industry continues to be major contributor to the diets of Americans and the economic viability of rural communities as well as for many states. The long-term goal of the overall project is to improve the efficiency of milk production, cow health and longevity, and thus promote continued food sufficiency, environmental sustainability, and economic sustainability in the PA US dairy industries. Our approach to achieve this goal is to systematically identify those biological and nutritional management processes that will provide the greatest improvement and to concentrate our research efforts there. The team will work on 2 of the major objectives:Objective 1: To quantify factors that impact supply and availability of nutrients utilized for efficient milk production while reducing environmental impactObjective 2: To identify and quantify molecular, cellular, and organismal signals that regulate intake, partitioning and efficient utilization of nutrientsThe team will conduct experiments to determine the possibility of maintaining high milk production and milk composition in dairy cows fed metabolizable protein (MP)-deficient diets by supplementing limiting essential amino acids. The role and requirements of digestible histidine in diets that meet lysine and methionine requirements will be established in a series of experiments with lactating dairy cows. Experiments will also be conducted to determine the role of individual amino acids in ruminal microbial protein synthesis and composition. A novel in vivo method to measure rumen biohydrogenation will be used to measure biohydrogenation of different unsaturated fatty acids and the impact of potential modifiers of the rumen microbiome on biohydrogenation. Factors limiting fatty acid digestibility in the lactating dairy cow will be studied. The effect of duodenal fatty acid profile on digestibility will be investigated using in vitro and in vivo approaches to identify key limiting points of emulsification and absorption. The effect of short and long chain fatty acid supply on milk fat synthesis will be studied in experiments with lactating dairy cows. Experiments will determine the effect of fatty acid supplements on mammary metabolism and milk fat synthesis pathways.

Animal Health Component

70%

Research Effort Categories

Basic

30%

Applied

70%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
302	3410	1010	100%

Knowledge Area
302 - Nutrient Utilization in Animals;

Subject Of Investigation
3410 - Dairy cattle, live animal;

Field Of Science
1010 - Nutrition and metabolism;

Keywords

dairy cattle

amino acid

protein

fatty acid

rumen biohydrogenation

fatty acid metabolism

Goals / Objectives
Objective 1: To quantify factors that impact supply and availability of nutrients utilized for efficient milk production while reducing environmental impact Objective 2: To identify and quantify molecular, cellular, and organismal signals that regulate intake, partitioning and efficient utilization of nutrients

Project Methods
The possibility of maintaining high milk production and milk composition in dairy cows fed metabolizable protein (MP)-deficient diets by supplementing limiting essential amino acids will be studied. The role and requirements of digestible histidine in diets that meet lysine and methionine requirements will be established in a series of experiments with lactating dairy cows. The first 2 experiments will be Latin square with 16 cows each: Exp. 1 will be with MP-adequate basal diet and treatments will be 4 levels of digestible histidine (from 1.8 to 3.0% of MP); Exp. 2 will have similar design but the basal diet will meet the MP requirements of the cows. The 3rd experiment will be a production type and dry matter intake will be monitored using the Calan gates system. Treatments in Exp. 3 will be 1 or 2 digestible histidine levels (plus a control), established in Exp. 1 and 2. In all experiments, rumen-protected methionine and lysine will be supplemented to the basal diet to meet the requirements of these 2 limiting amino acids. Experiments will also be conducted to determine the role of individual amino acids in ruminal microbial protein synthesis and composition. This will be done in a series of in vitro experiments in which 3 concentrations of all essential and some non-essential amino acids will be studied. Responses will include, rumen fermentation variables (pH, volatile fatty acids, ammonia, and total gas, methane, and hydrogen production), neutral-detergent fiber degradability, and microbial protein synthesis. It is expected that a total of 12 in vitro experiments will be conducted. A novel in vivo method to measure rumen biohydrogenation has been developed in the Harvatine lab and will be used to measure biohydrogenation of different unsaturated fatty acids and the impact of potential modifiers of the rumen microbiome on biohydrogenation. Secondly, the impact of rumen available methionine on biohydrogenation will be investigated in lactating dairy cattle. Another project will investigate factors limiting fatty acid digestibility in the lactating dairy cow. Fat supplements containing low levels of unsaturated fatty acids have poor digestibility. The effect of duodenal fatty acid profile on digestibility will be investigated using in vitro and in vivo approaches to identify key limiting points of emulsification and absorption. The effect of short and long chain fatty acid supply on milk fat synthesis will be studied in vivo in experiments with lactating dairy cows. Acetate and palmitic acid have been shown to increase milk fat yield in some instances. However, the mechanism is not clear. Experiments will be conducted to determine the effect of these supplements on mammary metabolism and milk fat synthesis pathways.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Animal scientists, Animal Science undergraduate and graduate students, extension, general public Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This research included the training of one MS and three PhD students.Research experience for 13 undergraduate students including two undergraduate honors thesis projects were also included. How have the results been disseminated to communities of interest?Peer-reviwed journal publications and scientific presentations What do you plan to do during the next reporting period to accomplish the goals?We will conduct a long-term, continuous design experiment to investigate the interaction of metabolizable protein level with digestible histidine supply in dairy cow diets. This work will help us formulate histidine requirements for dairy cows fed corn silage-based diets. We will continue our work to screen and identify seaweed species with a potential to decrease enteric methane emission in dairy cows without compromising nutrient digestibility or lactational performance. We will also continue studying the impact of methane inhibitors on the rumen microbiota. We will continue to investigate the effect of acetate on milk fat synthesis including the interaction with DGAT1 genotype of cows and interaction with high starch and unsaturated fat diets. We will also investigate the effect of high oleic soybeans on milk fat synthesisand fatty acid digestibility. Methods to improve fatty acid digestibility will also be investigated.

Impacts
What was accomplished under these goals? Objective 1: A. N. Hristov -Two experiments (MP-adequate and deficient diets) were conducted to determine dose effect of histidine (His) on milk production and composition and plasma amino acid (AA) concentrations in lactating dairy cows fed a metabolizable protein (MP) adequate or deficient diets. Milk and milk protein yields were optimized with digestible His supply at around 3.0% of MP in both experiments, with the effect of supplemental His being more pronounced with the MP-deficient diet. In a study with transition cows, we observed that plasma concentration of essential AA decreased after calving, indicating an increased demand for AA around parturition. Multiparous cows had lower postpartum plasma concentration of AA, likely reflecting an increased need for AA to sustain greater milk production compared with primiparous cows. In another study, 130 Holstein cows were screened for daily enteric methane emission and it was found that phenotypically low-methane yield cows had lower methane emission intensity, greater molar proportion of propionate and lower A:P ratio, and higher proportion of Methanosphaera vs. Methanobrevibacter in ruminal contents than phenotypically high-methane yield cows. A seaweed screening project identified species with potent methane-mitigating effect. Asparagopsis taxiformis (AT) fed at 0.5% of DMI decreased methane emission by ≥ 55% but also decreased DMI and milk production. A meta-analysis of Penn State's data showed that a methane inhibitor, 3-NOP, decreased daily enteric methane emission by an average of 24% and both methane yield and intensity by 25% without affecting dry matter intake or milk yield, and increased milk fat concentration in lactating dairy cows. Another experiment tested the hypothesis that increased oleic acid levels in a saturated fatty acid supplement reacted with magnesium would increase milk yield without induction of milk fat depression. The study showed that fat supplements tended to decrease milk protein in multiparous cows and that replacing a constant ratio of C16:0 to C18:0 with oleic acid does not negatively impact milk yield and milk fat yield. A study with roasted high oleic soybeans (Plenish, Pioneer Inc. Johnston, IA) demonstrated that the Plenish soybeans maintained milk and milk component yield when fed at up to 15% of the diet without induction of diet-induced milk fat depression. A study designed to investigate the effect of methionine on milk fat depression concluded that both rumen available 2-hydroxy-4-(methylthio)butanoate analog and rumen protected methionine decreased risk of milk fat depression. Objective 2: K. Harvatine -We investigated the effect of time of acetate absorption on the daily rhythm of milk synthesis and plasma hormones and metabolites in dairy cows. When compared to acetate infusion over 22 h/d (control), infusion over 8 h/d from 0900 to 1700 h increased the amplitude of plasma acetate by 129%, and infusion from 2100 to 0500 h decreased the amplitude by 28%, and both treatments result in a phase shift. The rhythm of other variables of secondary interest were also affected by treatment. These results support a role of the timing of acetate absorption in entraining the daily rhythm of milk synthesis. Another study investigated if glucose tolerance and insulin sensitivity follow a 24 h rhythm in dairy cows. The conclusion from the study was that glucose tolerance follows a daily rhythm in dairy cows, with greater glucose tolerance in morning and afternoon compared to overnight.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Hristov, A. N., and A. Melgar. 2020. Short communication: Relationship of dry matter intake with enteric methane emission measured with the GreenFeed system in dairy cows receiving a diet without or with 3-nitrooxypropanol. Animal 14(S3):s484s490. doi:10.1017/S1751731120001731.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Raisanen, S. E., C. M. M. R. Martins, K. Nedelkov, J. Oh, M. T. Harper, A. Melgar, X. Chen, C. Parys, R. A. Patton, M. Miura, and A. N. Hristov. 2020. Bioavailability of rumen-protected methionine, lysine and histidine assessed by fecal free amino acid excretion. Anim. Feed Sci. Technol. 268: 114595 https://doi.org/10.1016/j.anifeedsci.2020.114595.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Melgar, A., K. C. Welter, K. Nedelkov, C. M. M. R. Martins, M. T. Harper, J. Oh, S. E. Raisanen, X. Chen, S. F. Cueva, S. Duval, and A. N. Hristov. 2020. Dose-response effect of 3-nitrooxypropanol on enteric methane emission in dairy cows. J. Dairy. Sci. 103:61456156.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Melgar, A., M. T. Harper, J. Oh, F. Giallongo, M. E. Young, T. L. Ott, S. Duval, and A. N. Hristov. 2020. Effects of 3-nitrooxypropanol on rumen fermentation, lactational performance, and the resumption of ovarian cyclicity in dairy cows. J. Dairy. Sci. 103:410432.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Hristov, A. N., M. T. Harper, G. Roth, C. Canale, and P. Huhtanen. 2020. Effect of ensiling time on corn silage neutraldetergent fiber degradability and relationship between laboratory fiber analyses and in vivo digestibility. J. Dairy. Sci. 103:23332346. DOI: https://doi.org/10.3168/jds.2019-16917.
• Type: Journal Articles Status: Awaiting Publication Year Published: 2020 Citation: Shepardson, R.P., E. Bazileskaya, and K.J. Harvatine. 2020. Physical characterization of fatty acid supplements with varying enrichments of palmitic and stearic acid by differential scanning calorimetry. J. Dairy Sci. In Press.
• Type: Journal Articles Status: Awaiting Publication Year Published: 2020 Citation: Matamoros, C., R. Klopp, L.E. Moraes, and K.J. Harvatine. 2020. Meta-analysis of the relationship between milk trans-10 C18:1, milk fatty acids < 16 C, and milk fat production. J. Dairy Sci. In Press.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Andreen, D.M., M.M. Haan, C.D. Dechow, and K.J. Harvatine. 2020. Relationships between milk fat and rumination time recorded by commercial rumination sensing systems. J. Dairy Sci. 103:8094-8104.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Salfer, I.J. and K.J. Harvatine. 2020. Night-restricted feeding of dairy cows modifies daily rhythms. Bri. J. Nutr. 123:849-858.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Pitta, D.W., N. Indugu, B. Vecchiarelli, M. Hennessy, M. Baldin, and K.J. Harvatine. 2020. Effect of 2-hydroxy-4-(methylthio) butanoate (HMTBa) supplementation on rumen bacterial populations in dairy cows when exposed to diets with risk for milk fat depression. J. Dairy Sci. 103:2718-2730.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Salfer, I.J., P.A. Bartell, C.D. Dechow, and K.J. Harvatine. 2020. Annual rhythms of milk synthesis in dairy herds in four regions of the United States and their relationships to environmental predictors. J. Dairy Sci. 103:3696-3707.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Nedelkov, K., X. J. Chen, C.M.M.R. Martins, A. Melgar, M.T. Harper, S. R�is�nen, J. Oh, T. L. Felix, E. Wall, and A. N. Hristov. 2020. Short communication: Alternative selenium supplement for sheep. Anim. Feed Sci. Technol. 261:114390. https://doi.org/10.1016/j.anifeedsci.2020.114390
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Chen, X., K. Nedelkov, J. Oh, M. T. Harper, E. H. Wall, and A. N. Hristov. 2019. Effect of a blend of an artificial sweetener and capsicum on productive performance and blood chemistry in lambs. Anim. Feed Sci. Technol. https://doi.org/10.1016/j.anifeedsci.2019.114308
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Robblee, M., Y.R. Boisclair, D.E. Bauman, and K.J. Harvatine. 2019. Dietary fat does not overcome trans-10, cis-12 conjugated linoleic acid (CLA) inhibition of milk fat synthesis in lactating mice. Lipids. 55:201-212
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Andreen, D.M., I.J. Salfer, Y. Ying, D.J. Reinemann, and K.J. Harvatine. 2019. Technical Note: Method for improving precision of in-parlor milk meters and adjusting milk weights for stall effects. J. Dairy Science. 103:5162-5169

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Animal scientists, Animal Science undergraduate and graduate students, Extension, general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Attendance of ADSA, Discover, and international meetings for the scientists and graduate students on the project. How have the results been disseminated to communities of interest?Yes, through publications in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?We will continue working on current projects to address Objectives 1 and 2 of the project.

Impacts
What was accomplished under these goals? Under Objective 1 "To quantify factors that impact supply and availability of nutrients utilized for efficient milk production while reducing environmental impact", we investigated the effect of a methane (CH4) inhibitor, of 3-nitrooxypropanol (3NOP), on enteric methane emission and lactational performance of Holstein dairy cows. Compared with the control, 3NOP decreased (P < 0.001) CH4 daily emission by 26% (302 vs. 411 g/d; SEM = 6.16); decreased (P < 0.001) CH4 emission yield (11.9 vs. 16.4 g/kg DMI; SEM = 0.25), and emission intensity [8.2 vs. 11.5 g/kg energy corrected milk (ECM); SEM = 0.28]. Treatment had no effect (P ≥ 0.45) on milk and ECM yields, averaging 38.4 kg (SEM= 0.63) and 37.2 kg/d, (SEM = 0.59), respectively. ECM feed efficiency was also not affected (P ≥ 0.94) by 3NOP. In this experiment, 3NOP decreased enteric CH4 daily emission, yield, and intensity without affecting DMI and milk yield, but increased milk fat in lactating dairy cows. Further, an analysis of CH4 emission data from a large experiment showed that the relationship of enteric CH4 emission, measured using the GreenFeed system, and DMI in dairy cows depends on the time of measurement relative to time of feeding and can be as high as determined in respiration chambers. We found that the methane mitigation effect of 3-NOP is highest immediately after feeding and lowest before feeding. In another study, we quantified variation in rumination time between and within dairy herds and to determine relationships between rumination time and milk fat production and fatty acid (FA) profile as a proxy of rumen fermentation. While rumination was not related directly to milk fat, it was associated with differences in trans and odd and branched-chain FA that also change during SARA or MFD, which may impact milk fat and other production variables. Further investigation using rumination data from commercial systems to predict or identify the presence of these conditions is warranted. In yet another study, we investigated if the effect of NaAcet on milk fat production is due to increasing acetate supply or DCAD. Milk yield was not different between treatments, but NaAcet and NaHCO3 increased dry matter intake by 6.6 and 7%, respectively (P = 0.02). Milk fat yield was increased 134 g by NaAcet and 118 g by NaHCO3 (P <0.01). Sodium acetate increased milk fat yield predominantly by increasing the yield of de novo and mixed source fatty acids, while NaHCO3 increased the yield of preformed and de novo fatty acids. Sodium acetate increased plasma acetate and decreased plasma glucose concentration during the afternoon and early evening, which coincides with the higher intake period of the day. In conclusion, NaAcet and NaHCO3 both increase milk fat production, but NaAcet achieves this through increased mammary gland de novo lipogenesis. Under Objective 2: To identify and quantify molecular, cellular, and organismal signals that regulate partitioning and efficient conversion of nutrients to milk" we conducted a study with the objective to determine the effects of the timing of protein absorption on the daily rhythms of milk synthesis in dairy cows. Nine lactating Holstein cows were randomly assigned to 1 of 3 treatment sequences in a 3 x 3 Latin square. Treatments were absomasal infusions of 500 g/d of sodium caseinate either 24 h/d (CON) or for 8 h/d from 0900 to 1700 h (DAY) or from 2100 to 0500 (NGT). The time of protein infusion influenced daily rhythms of milk and milk protein synthesis. Night infusion abolished rhythms of protein concentration and induced rhythms of milk yield. Day infusion increased the amplitude of protein concentration. The results of this study suggest a role of amino acids in entraining the molecular clock of the mammary gland. Another study was conducted to determine the effects of the timing of fatty acid absorption on the daily rhythms of milk synthesis. We concluded that fatty acid infusion during the daytime modified the daily rhythms of milk synthesis by increasing the amplitude of milk yield and decreasing the amplitude of fat and protein concentration, whereas infusion at night had little effect.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Harper, M. T., J. Oh, A. Melgar, K. Nedelkov, S. R�is�nen, X. Chen, C. M. M. R. Martins, M. Young, T. L. Ott, D. M. Kniffen, R. A. Fabin, and A. N. Hristov. 2019. Production effects of feeding extruded soybean meal to early-lactation dairy cows. J. Dairy. Sci. 102 (in press); doi.org/10.3168/jds.2019-16551
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Oh, J., M. Harper, A. Melgar, D. M. Paulus Compart, and A. N. Hristov. 2019. Effect of Saccharomyces cerevisiae-based direct-fed microbial and exogenous enzyme products on enteric methane emission and productivity of lactating dairy cows. J. Dairy. Sci. 102:60656075.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Hristov, A. N., A. Bannink, L. A. Crompton, P. Huhtanen, M. Kreuzer, M. McGee, 12 P. Nozi�re, C. K. Reynolds, A. R. Bayat, D. R. Y��ez-Ruiz, J. Dijkstra, E. Kebreab, A. Schwarm, K. J. Shingfield, and Z. Yu. 2019. Nitrogen in ruminant nutrition: a review of measurement techniques. J. Dairy Sci. 102:58115852.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Salfer, I.J., C.D. Dechow, and K.J. Harvatine. 2019. Annual rhythms of milk and milk fat and protein production in dairy cattle in the United States. J. Dairy Sci. 102:742-753.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Urrutia, N., R. Bomberger, C. Matamoros, and K.J. Harvatine. 2019. Effect of dietary supplementation of sodium acetate and calcium butyrate on milk fat synthesis in lactating dairy cows. J. Dairy Sci. 102:5172-5181.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Baldin, M., H.A. Tucker, and K.J. Harvatine. 2019. Milk fat response and milk fat and urine marker prediction of biomarkers of microbial nitrogen flow during supplementation with 2-hydroxy-4-(methylthio)butanoate (HMTBa). J. Dairy Sci. 102:6157-6166.