Tierärztl. Hochsch., Diss., 2003--Hannover.

Berlin, Humboldt-Universität, Thesis (doctoral), 2001.

Knowledge on the precise effects of surplus glucose supply in dairy cows is limited by the lack of information on how intermediary metabolism adapts at different levels of glucose availability. Therefore, a gradual increase of glucose supply via intravenous glucose infusion was used in the present study to test the dose effect of surplus provision of glucose on the metabolic status and milk production of dairy cows. Furthermore, the effects of increasing levels of surplus glucose on mRNA expressions and activities of rate-limiting enzymes involved in hepatic gluconeogenesis were investigated. Based on a previous finding that a positive energy balance may decrease hepatic carnitine palmitoyltransferase (CPT) enzyme activity, it was also of interest whether skeletal muscle CPT activity is downregulated in a similar manner during positive energy balance. Twelve midlactating Holstein-Friesian dairy cows were continuously infused over a 28-d experimental period with either saline (SI group, six cows) or 40% glucose solutions (GI group, six cows). The infusion dose was calculated as a percentage of the daily energy (NEL) requirements by the animal, starting at 0% on d 0 and increasing gradually by 1.25%/d until a maximum dose of 30% was reached by d 24. Dose was then maintained at 30% NEL requirement for 5 d. No infusions were made between d 29-32. Liver and skeletal muscle biopsies were taken on d 0, 8, 16, 24, and 32. Body weight (BW) and back fat thickness (BFT) were recorded on biopsies days. Blood samples were taken every 2 d. In addition, blood samples over 24 h (6-h intervals) were taken the days before each biopsy. Milk and urine samples were taken on biopsies days. BW and BFT increased linearly with increasing glucose dose for GI cows. No differences were observed in the dry matter intake, milk energy output, and energy corrected milk yield between groups. However, milk protein percentage and yield increased linearly in the GI group. Only occasional increases in blood glucose and insulin concentrations were observed in blood samples taken at 1000 h every 2 d. However, during infusion dose of 30% NEL requirements on d 24, GI cows developed postprandial hyperglycemia associated with hyperinsulinemia, coinciding with glucosuria. The revised quantitative insulin sensitivity check index (RQUIKI) indicated linear development of insulin resistance for the GI treatment. GI decreased serum concentrations of beta-hydroxybutyrate (BHBA) and blood urea nitrogen and tended to decrease the serum concentration of non-esterified fatty acids (NEFA). Liver glycogen content increased, while glycogen content in skeletal muscle only tended to increase by GI. No significant changes were observed in the activities and relative mRNA expression levels of hepatic phosphoenolpyruvate carboxykinase and glucose 6-phospatase. The activity of fructose 1,6-bisphosphatase (FBPase) and relative mRNA expression levels of pyruvate carboxylase (PC) were decreased in the GI group but only during the high dose of glucose infusion. Hepatic CPT activity decreased with GI and remained decreased on d 32. The hepatic expression levels of CPT-1A and CPT-2 mRNA were not significantly altered but tended to reflect the changes in enzyme activity. No effect of glucose infusion was observed on skeletal muscle CPT activity. The aforementioned adaptations were reversed four days after the end of glucose infusions except for those of BW, BFT, and lipid metabolism (i.e. serum BHBA and NEFA concentrations, hepatic CPT activity). It is concluded that mid-lactation dairy cows on an energy-balanced diet direct intravenously infused glucose predominantly to body fat reserves but not to increased lactation performance. Cows rapidly adapted to increasing glucose supply but experienced dose-dependent development of insulin resistance corresponding with postprandial hyperglycemia/hyperinsulinemia and glucosuria at dosages equivalent to 30% NEL requirements. The catalytic capacity of key hepatic gluconeogenesis enzymes in mid-lactating dairy cows is not significantly affected by nutritionally relevant increases of glucose supply. Only very high dosages selectively suppress PC transcription and FBPase activity. Finally, it can be concluded that suppression of CPT activity by positive energy balance appears to be specific for the liver in midlactating dairy cows.

Knowledge on the precise effects of surplus glucose supply in dairy cows is limited by the lack of information on how intermediary metabolism adapts at different levels of glucose availability. Therefore, a gradual increase of glucose supply via intravenous glucose infusion was used in the present study to test the dose effect of surplus provision of glucose on the metabolic status and milk production of dairy cows. Furthermore, the effects of increasing levels of surplus glucose on mRNA expressions and activities of rate-limiting enzymes involved in hepatic gluconeogenesis were investigated. Based on a previous finding that a positive energy balance may decrease hepatic carnitine palmitoyltransferase (CPT) enzyme activity, it was also of interest whether skeletal muscle CPT activity is downregulated in a similar manner during positive energy balance. Twelve midlactating Holstein-Friesian dairy cows were continuously infused over a 28-d experimental period with either saline (SI group, six cows) or 40% glucose solutions (GI group, six cows). The infusion dose was calculated as a percentage of the daily energy (NEL) requirements by the animal, starting at 0% on d 0 and increasing gradually by 1.25%/d until a maximum dose of 30% was reached by d 24. Dose was then maintained at 30% NEL requirement for 5 d. No infusions were made between d 29-32. Liver and skeletal muscle biopsies were taken on d 0, 8, 16, 24, and 32. Body weight (BW) and back fat thickness (BFT) were recorded on biopsies days. Blood samples were taken every 2 d. In addition, blood samples over 24 h (6-h intervals) were taken the days before each biopsy. Milk and urine samples were taken on biopsies days. BW and BFT increased linearly with increasing glucose dose for GI cows. No differences were observed in the dry matter intake, milk energy output, and energy corrected milk yield between groups. However, milk protein percentage and yield increased linearly in the GI group. Only occasional increases in blood glucose and insulin concentrations were observed in blood samples taken at 1000 h every 2 d. However, during infusion dose of 30% NEL requirements on d 24, GI cows developed postprandial hyperglycemia associated with hyperinsulinemia, coinciding with glucosuria. The revised quantitative insulin sensitivity check index (RQUIKI) indicated linear development of insulin resistance for the GI treatment. GI decreased serum concentrations of beta-hydroxybutyrate (BHBA) and blood urea nitrogen and tended to decrease the serum concentration of non-esterified fatty acids (NEFA). Liver glycogen content increased, while glycogen content in skeletal muscle only tended to increase by GI. No significant changes were observed in the activities and relative mRNA expression levels of hepatic phosphoenolpyruvate carboxykinase and glucose 6-phospatase. The activity of fructose 1,6-bisphosphatase (FBPase) and relative mRNA expression levels of pyruvate carboxylase (PC) were decreased in the GI group but only during the high dose of glucose infusion. Hepatic CPT activity decreased with GI and remained decreased on d 32. The hepatic expression levels of CPT-1A and CPT-2 mRNA were not significantly altered but tended to reflect the changes in enzyme activity. No effect of glucose infusion was observed on skeletal muscle CPT activity. The aforementioned adaptations were reversed four days after the end of glucose infusions except for those of BW, BFT, and lipid metabolism (i.e. serum BHBA and NEFA concentrations, hepatic CPT activity). It is concluded that mid-lactation dairy cows on an energy-balanced diet direct intravenously infused glucose predominantly to body fat reserves but not to increased lactation performance. Cows rapidly adapted to increasing glucose supply but experienced dose-dependent development of insulin resistance corresponding with postprandial hyperglycemia/hyperinsulinemia and glucosuria at dosages equivalent to 30% NEL requirements. The catalytic capacity of key hepatic gluconeogenesis enzymes in mid-lactating dairy cows is not significantly affected by nutritionally relevant increases of glucose supply. Only very high dosages selectively suppress PC transcription and FBPase activity. Finally, it can be concluded that suppression of CPT activity by positive energy balance appears to be specific for the liver in midlactating dairy cows.