Final Report for SAIF Project: Differences in Vitamin C Concentrations between High versus Low-producing Dairy Cows

Transcription

1 Final Report for SAIF Project: Differences in Vitamin C Concentrations between High versus Low-producing Dairy Cows James H. Hampton, School of Agriculture 1) Summary of Research and Results: See attached documents 2) Benefit to the University Community There were several beneficial outcomes to this work. One would be the scientific value of the data itself. Vitamin C concentrations have been poorly studied in lactating cattle, primarily due to the assumption that ruminants have sufficient capacity to produce their own Vitamin C. Since the time when vitamin C was first examined in ruminants, assay sensitivities have improved to be able to detect minute differences that were previously undetectable in ruminants. Results from this experiment could lead to changes in vitamin supplementation recommendations in lactating animals and potentially improve milk production in dairy cattle. A second, and almost greater benefit in my opinion, has been the inclusion and participation of a student in undergraduate research. The student assisting me gained valuable skills in experimental design, data collection, laboratory analysis, and general scientific methods and techniques. The student also was able to apply and integrate information learned in previous course work and was able to share this information in a variety of venues. One hundred percent of this SAIF funding was used to purchase supplies and equipment to support this project. 3) Dissemination of Results Oral, written, and poster presentations of results were prepared by the undergraduate student working on the project, Alecia Knapp. Alecia first presented preliminary data in Fall of 2014 in ANSCI 4190 Seminar in Animal Science as an oral class presentation. Alecia then presented final data, in poster format (see attached), at both the 2015 Posters in the Rotunda event in Madison, WI and on campus at our Spring 2015 URCE Symposium. Alecia also prepared a final written report to fulfil her ANSCI 4980 Undergraduate Research requirements (see attached). This work has also been shared in my ANSCI 3110 Reproductive Physiology of Domestic Animals course and will serve as a foundation for continuing research in this area.

2 1 Difference in the levels of Vitamin C between high versus low Producing Dairy Cows Alecia P. Knapp and J.H. Hampton University of Wisconsin-Platteville Animal Science Department Abstract We hypothesize that high-producing dairy cows will have lower Vitamin C concentrations versus low-producing dairy cows due to their bodies higher demand for glucose. Lactating Holstein cows (n=40) were randomly selected from the University of Wisconsin Platteville milking herd and assigned to either high production (HP; n=20) or low-production (LP; n=20) group based on DHI predicted 305-day milk yields (Ag Cooperative Services). Cows in the HP group had predicted 305-day milk yields 27,000 lbs. of milk while cows in the LP group had predicted 305-day milk yields <26,999 lbs. of milk. Neither number of days in milk (DIM) or lactation number were significant between groups (P =.3041; P=.1983). Serum samples were used to measure Vitamin C concentrations using the Cayman Chemical Ascorbate Assay kit. The average Vitamin C concentration for the HP group was 17.87±1.03 and the LP group was 31.83±1.03 (P <0.0001). Results support previous studies, which showed that lactating cows may not have enough dietary glucose to produce milk and also produce enough Vitamin C (MacLeod, 1999). Further studies are necessary to determine if supplementation of glucose increases Vitamin C concentrations in high producing dairy cows. Introduction Vitamin C (C 6 H 8 O 6 ) is a water soluble vitamin with antioxidant properties. Some species, such as humans, cannot manufacture their own, while others, including ruminants, can perform biosynthesis of vitamin C. Vitamin C biosynthesis in ruminants occurs in the liver and process

3 2 glucose. Because ruminants are able to produce their own vitamin C a dietary requirement has yet to be determined. It is important that ruminants produce their own vitamin C as the bacteria in the rumen break down a high majority of any dietary supplements of the vitamin. Glucose is a precursor to both vitamin C and milk, so it is in high demand in the body of a lactating dairy cow. Studies have shown that when a high producing dairy cow is lactating, her body allocates glucose to milk production over that of Vitamin C, which can lead to a shortage of Vitamin C. A lactating dairy cow s body is under a high level of stress, and antioxidants, such as vitamin C, help to manage that stress level. When there is a shortage of vitamin C this reduces the body s ability to handle the stress from lactating, which can be measured via cortisol concentrations. Cortisol is a glucocorticoid produced by the bovine adrenal gland and is released in response to chronic stress. Cortisol increases coincident with peak lactation (40-60 days postcalving) in dairy cows (Sartin et al., 1988) indicating that during this time period cattle are under greater stress. Previous studies have indicated that Vitamin C, an antioxidant, has the ability to inhibit cortisol production (Montalvo et al., 2001). Thus, the effect of reduced vitamin C is amplified in high-producing versus low-producing dairy cows, since high producing dairy cows have higher blood concentrations of cortisol (Sartin et al., 1988). We hypothesize that high-producing dairy cows will have lower Vitamin C concentrations versus low-producing dairy cows due to their bodies higher demand for glucose. Materials and Methods All procedures were approved by the University of Wisconsin-Platteville Animal Care and Use Committee. Lactating Holstein cows (n=40) were randomly selected from the

4 3 University of Wisconsin Platteville milking herd and assigned to either high production (HP; n=20) or low-production (LP; n=20) group based on DHI predicted 305-day milk yields (Ag Cooperative Services). Cows in the HP group had predicted 305-day milk yields 27,000 lbs. of milk while cows in the LP group had predicted 305-day milk yields <26,999 lbs. of milk. Number of days in milk (DIM) ranged from , and lactation number ranged from 1st to 4th lactation. All cattle were housed at the University of Wisconsin-Platteville Dairy Center in 2 adjacent free-stall pens and were provided a balanced TMR, free choice mineral mix, and water. Blood was collected from the coccygeal vein at the underside of the base of the tail. All samples were collected after morning milking, on the same day within four hours of each other. SST 8.5 ml BD Vacutainers with clot activator were used. After collection the blood was allowed to clot for approximately 30 minutes and then placed on ice. Samples were then centrifuged within two hours of collection for 30 minutes at 1800 G. Serum was then separated and put into deep well plates. Protocol was then followed to the point of sample storage for the Cayman Chemical Ascorbate Assay kit. Samples were then stored at -80 C until they were shipped to the Pennington Biomedical Research Center in Baton Rouge, Louisiana for the completion of the assay. Results All statistical measures were performed using SAS. Average DIM = 102 ± 7 days, and was not significantly different between groups (P=0.3041). Average lactation number = 2.1±0.2, and was not significantly different between groups (P=0.1983). Predicted 305-day milk averages were 31,887± lbs. of milk (HP) and 23,155± lbs. of milk (LP) and were

5 4 significantly different (P <0.0001). Vitamin C concentration averages were significantly lower in the HP group (17.87±1.03) than the LP group (31.83±1.03) with P< (Figure 1). 35 Serum Ascorbic Acid Concentrations (µm) of High vs Low Producing Dairy Cows Serum Ascorbic Acid Concentration (um) High Production Level Low Figure 1: Serum Vitamin C levels in Lactating Dairy Cows with. High Production Mean =17.87 ±1.03. Low Production Mean = ±1.03 (P <0.0001) Discussion Our results support the hypothesis that high producing dairy cows have a lower vitamin C due to reduced glucose availability. When both high and low producing dairy cows are feed the same ration, the lower producing cows will use less of the glucose gained from their diet for milk, leaving more left over for vitamin C production and other biological processes. Further

6 5 research will need to be done comparing cortisol levels in high and low producing dairy cows to see if there is a correlation with the vitamin C levels. Also, research can be done looking at glucose supplementation and vitamin C levels in lactating dairy cows. References MacLeod, D. D., X. Zhang, L. Ozimeck and J. J. Kennelly. Ascorbyl-2-polyphosphate as a source of ascorbic acid for dairy cattle. Milchwissenschaft V54: Montalvo, CP, NH Díaz, LA Galdames, ME Andrés, and RE Larraín. Short communication: effect of vitamins E and C on cortisol production by bovine adrenocortical cells in vitro. Journal of Dairy Science V 94(7): Sartin, JL, RJ Kemppainen, KA Cummins, and JC Williams. Plasma Concentrations of Metabolic Hormones in High and Low Producing Dairy Cows. Journal of Dairy Science V71 (3): Acknowledgements Dr. James Hampton, Research and Academic Advisor Dr. Christopher Morrison, Pennington Biomedical Research Center Provost Nimocks Den Herder UW-Platteville Department of Animal Science Krystan Scheffler, Assistant Chris and Shannon Knapp, Parents Office of Undergraduate Research and Creative Endeavors, Support Pioneer Undergraduate Research Fellow Program, Funding

7 U N I V E R S I T Y O F W I S C O N S I N - P L A T T E V I L L E O F F I C E O F U N D E R G R A D U AT E RESEARCH A N D C R EAT I V E ENDEAVORS Difference in the levels of Vitamin C between high versus low Producing Dairy Cows Alecia P. Knapp and J.H. Hampton Materials & Methods Abstract Discussion All procedures were approved by the University of Wisconsin- Platteville Animal Care and We hypothesize that high- producing dairy cows will have higher Vitamin C concentrations Our results support the hypothesis that high producing dairy cows have a lower vitamin C versus low- producing dairy cows which will better enable them to combat high stress- induced Use Committee. Lactating Holstein cows (n=40) were randomly selected from the University of Wisconsin due to reduced glucose availability. When both high and low producing dairy cows are feed the same ration, concentrations of cortisol. Lactating Holstein cows (n=40) were randomly selected from the University of Platteville milking herd and assigned to either high production (HP; n=20) or low- production (LP; n=20) the lower producing cows will use less of the glucose gained from their diet for milk, leaving more left over Wisconsin Platteville milking herd and assigned to either high production (HP; n=20) or low- production (LP; group based on DHI predicted 305- day milk yields (Ag Cooperative Services). Cows in the HP group had for vitamin C production and other biological processes. Further research will need to be done comparing n=20) group based on DHI predicted 305- day milk yields (Ag Cooperative Services). Cows in the HP group predicted 305- day milk yields 27,000 lbs. of milk while cows in the LP group had predicted 305- day milk cortisol levels in high and low producing dairy cows to see if there is a correlation with the vitamin C levels. had predicted 305- day milk yields 27,000 lbs. of milk while cows in the LP group had predicted 305- day yields <26,999 lbs. of milk. Number of days in milk (DIM) ranged from , and lactation number ranged Also, research can be done looking at glucose supplementation and vitamin C levels in lactating dairy cows. milk yields <26,999 lbs. of milk. Neither number of days in milk (DIM) or lactation number were significant from 1st to 4th lactation. All cattle were housed at the University of Wisconsin- Platteville Dairy Center in 2 between groups (P =.3041; P=.1983). Serum samples were used to measure Vitamin C concentrations using adjacent free- stall pens and were provided a balanced TMR, free choice mineral mix, and water. the Cayman Chemical Ascorbate Assay kit. The average Vitamin C concentration for the HP group was Blood was collected from the coccygeal vein at the underside of the base of the tail. All samples were 17.87±1.03 and the LP group was 31.83±1.03 (P <0.0001). Results support previous studies, which showed collected after morning milking, on the same day within four hours of each other. SST 8.5 ml BD that lactating cows may not have enough dietary glucose to produce milk and also produce enough Vitamin Vacutainers with clot activator were used. After collection the blood was allowed to clot for approximately C (MacLeod, 1999). Further studies are necessary to determine if supplementation of glucose increases 30 minutes and then placed on ice. Samples were then centrifuged within two hours of collection for 30 Vitamin C concentrations in high producing dairy cows. minutes at 1800 G. Serum was then separated and put into deep well plates. Protocol was then followed to Introduction Vitamin C (C6H8O6) is a water soluble vitamin with antioxidant properties. Some species, the point of sample storage for the Cayman Chemical Ascorbate Assay kit. Samples were then stored at - 80 C until they were shipped to the Pennington Biomedical Research Center in Baton Rouge, Louisiana for the completion of the assay. MacLeod, D. D., X. Zhang, L. Ozimeck and J. J. Kennelly. Ascorbyl- 2- polyphosphate as a source of ascorbic such as humans, cannot manufacture their own, while others, including ruminants, can perform acid for dairy cattle. Milchwissenschaft V54: biosynthesis of vitamin C. Vitamin C biosynthesis in ruminants occurs in the liver and process glucose. Montalvo, CP, NH Díaz, LA Galdames, ME Andrés, and RE Larraín. Short communication: effect of vitamins E Because ruminants are able to produce their own vitamin C a dietary requirement has yet to be Serum Ascorbic Acid Concentrations (µm) of High vs Low Producing Dairy Cows determined. It is important that ruminants produce their own vitamin C as the bacteria in the rumen break Glucose is a precursor to both vitamin C and milk, so it is in high demand in the body of a lactating dairy cow. Studies have shown that when a high producing dairy cow is lactating, her body allocates glucose to milk production over that of Vitamin C, which can lead to a shortage of Vitamin C. A lactating dairy cow s body is under a high level of stress, and antioxidants, such as vitamin C, help to manage that stress level. When there is a shortage of vitamin C this reduces the body s ability to handle the stress from lactating, which can be measured via cortisol concentrations. Cortisol is a glucocorticoid produced by the bovine adrenal gland and is released in response to chronic stress. Cortisol increases coincident with peak lactation (40-60 days postcalving) in dairy cows (Sartin et al., 1988) indicating that during this time period cattle are under greater stress. Previous studies have indicated that Vitamin C, an antioxidant, has the ability to inhibit cortisol production (Montalvo et al., 2001). Thus, the effect of reduced vitamin C is amplified in high- producing versus low- producing dairy cows, since high producing dairy cows have higher blood concentrations of cortisol (Sartin et al., 1988). We hypothesize that high- producing dairy cows will have higher Vitamin C concentrations versus low- producing dairy cows which will better enable them to combat high stress- induced concentrations of cortisol. 35 Serum Ascorbic Acid Concentration (mm) down a high majority of any dietary supplements of the vitamin. References and C on cortisol production by bovine adrenocortical cells in vitro. Journal of Dairy Science V 94(7): Sartin, JL, RJ Kemppainen, KA Cummins, and JC Williams. Plasma Concentrations of Metabolic Hormones in High and Low Producing Dairy Cows. Journal of Dairy Science V71 (3): Acknowledgements Dr. James Hampton, Research and Academic Advisor Dr. Christopher Morrison, Pennington Biomedical Research Center 10 Provost Nimocks Den Herder 5 Department of Animal Science Krystan Scheffler, Assistant 0 High Production Level Low Figure 1: Serum Vitamin C levels in Lactating Dairy Cows with. High Production Mean =17.87 ±1.03. Low Production Mean = ±1.03 (P <0.0001) Chris and Shannon Knapp, Parents Office of Undergraduate Research and Creative Endeavors, Support Pioneer Undergraduate Research Fellow Program, Funding Results All statistical measures were performed using SAS. Average DIM = 102 ± 7 days, and was not Stu d e n t Name: Alecia K napp significantly different between groups (P=0.3041). Average lactation number = 2.1±0.2, and was not Hometo wn : Waverly, I A significantly different between groups (P=0.1983). Predicted 305- day milk averages were 31,887± lbs. of milk (HP) and 23,155± lbs. of milk (LP) and were significantly different (P <0.0001). Vitamin C Legislative Di s tri c t: As s e m b l y concentration averages were significantly lower in the HP group (17.87±1.03) than the LP group (31.83±1.03) with P< (Figure 1). Glucose to Vitamin C Pathway Di s tri c t 17, S e n ate Di s tri c t 49 Vitamin C Molecule