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a K k 0.26 0.2O 0.10 0.05 O.OO GLYCEROLIPID SYNTHESIS IN BOVINE FATTY LIVER 79 partus — free access 4 6 TIME (WEEKS) 4 6 TIME (WEEKS) — restricted -i 10 12 free access restricted 10 12 4 6 TIME (WEEKS) 4 6 TIME (WEEKS) free access restricted FIGURE 1 Ante- and postpartum hepatic triacylglycerol (TAG) (A], phospholipids (PL) (B), cholesterol (C) and glycogen (D) concentrations in cows with either restricted (n = 7) or free access to feed (n = 8 or 10) during the dry period. Values are depicted as means ±SEM.One wk before parturition, all cows had free access to feed. Significant diet effects: liver triacylglycerols, P < 0.001 (0.5-12 wk postpartum); liver phospholipids, P < 0.001 (0.5-12 wk postpartum). postpartum period, there were data from 7 control and 8 test cows. All statistical analyses (Steel and Torry 1981) were performed using a SYSTATcomputer pro gram, version 5.0 (Wilkinson 1990). Data within di etary treatments were checked for normal distribution (Kolmogorov-Smirnov test). Normally distributed data were subjected to a repeated one-way ANOVA to evaluate diet effects. Diet effects at selected time in tervals were identified using Student's t test or, when the data were not distributed normally, by the Mann- Whitney U test. When the data had a normal distri bution, homogeneity of variances was verified using Bartlett's test. The Pearson-correlation coefficient was calculated between individual serum total cholesterol and phospholipid concentrations. The two-sided level of statistical significance was preset at P < 0.05. RESULTS Performance, Daily dry matter intake in the pe riod 10-14 wk antepartum was 20.6 ±0.42 kg for the test group with free access to feed and 7 kg for the control group given a restricted amount of feed. Dur ing the 2 wk just before calving, the intake of the test cows was 17.5 ±1.52 kg/d. Average dry matter intake in the test group was 15.5 ±2.87 kg/d during the first 0.5 wk after parturition; in the control group it was 17.4 ±2.35 kg/d. In the period of 0-4 wk after par turition, average daily dry matter intake was 22.0 ±0.83 kg in the test group and also 22.0 ±1.43 kg in the control group. Average milk yields in the first wk of lactation were 30.6 ±1.96 kg/d and 30.7 ±2.49 kg/d for the control and the test group, respectively. During wk 2-12 of lactation, milk yields were 37.0 ±1.09 for the test group and 38.1 ±1.48 kg/d for the control group. During the dry period, body weight gain in the con trol group was 112 ±15.1 kg, and in the overfed test group it was 152 ±16.2 kg, the difference not being statistically significant (P = 0.128). Body weight loss after parturition, i.e., the difference between body weight measured immediately postpartum and that at 4 wk postpartum, was 79 ±9.3 kg for the control group and 114 ±11.0 kg for the test group (P = 0.035). Liver lipids and glycogen. Before calving, liver lipid concentrations did not differ between the groups. Liver triacylglycerol concentrations rose after calving, the rise being significantly greater (P < 0.001; 0.5-12 wk postpartum) in the test group with free access to feed when compared with the control group that had Downloaded from by guest on December 18, 2012

80 VAN DEN TOP ET AL. 2.00 1.50 1.00 O.SO 0.00 partus free access 468 TIME (WEEKS) restricted parlus free access 468 TIME (WEEKS) restricted 10 12 0. 0 468 TIME (WEEKS) FIGURE 2 Ante- and postpartum activities of hepatic micorosomal (A) and mitochondrial glycerolphosphate acyltransferase (GPAT) (B) and microsomal (C) and cytosolic phosphatidate phosphohydrolase (PAP) (D) in cows with either restricted (n = 7) or free access to feed (n = 8 or 10) during the dry period. Values are means ±SEM. been given a restricted amount of feed during the dry period (Fig. 1A). In the test vs. the restricted control group, postpartum hepatic phospholipid concentra tions were slightly, but signifi cantly lower (P < 0.001 ; 0.5-12 wk postpartum)(Fig. IB). After calving, liver phospholipid concentrations in both groups initially rose and then declined until 2 wk postpartum and rose again to a level that was constant between 3 and 12 wk postpartum. Postpartum hepatic cholesterol con centrations were not different in the two groups, but they rose after calving (Fig. 1C). Liver glycogen con centrations before calving were similar in both groups, but after calving they fell to lower values in the test group than in the restricted control group (P = 0.067; 0.5-12 wk after calving)(Fig. ID). Enzymes involved in hepatic glycerolipid syn thesis. Before parturition, the activities of the hepatic glycerolipid synthesizing enzymes were not signifi cantly different for the two groups. Microsomal GPAT activities had risen significantly (P = 0.006) at 0.5 wk after calving when compared with 0.5 wk before calv ing and tended to be lower (P = 0.095; 0.5-3 wk post partum) in the test group (Fig. 2A). Likewise, post partum mitochondrial GPAT activities were signifi cantly lower in the test group than in the control group (P = 0.015; 1-3 wk postpartum) (Fig. 2B). PAP activ ities in both the cytosol and the microsomes rose after calving (P < 0.015) when compared with prepartum values. After parturition, the initially elevated group mean cytosolic and microsomal PAP activities grad ually declined, the values being similar for the two groups (Fig. 2C and D). DGAT activities were higher (P < 0.001) at 0.5 wk after calving when compared with 0.5 wk before calving. From 1 to 3 wk after calv ing DGAT activities remained constant and then gradually declined. Postpartum DGAT activities did not differ significantly between the control and test groups (Fig. 3C). Cytosolic CPCT activities fell, and microsomal CPCT activities rose immediately after parturition (P = 0.001). From 1.5 wk after parturition, CPCT activities gradually returned to levels compa rable with antepartum values. During the postpartum period under study, CPCT activities did not differ be tween the two groups (Fig. 3A and B). Serum indicators of liver function and cell damage. Prepartum activities of serum lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase did not significantly differ between the groups. After parturition, the activities of lactate dehydrogenase were significantly higher (P = 0.029; 0.5- 12 wk postpartum) in the test group (Fig. 4A). Alkaline phosphatase activities tended to be higher (P = 0.055; Downloaded from by guest on December 18, 2012

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