Activities of the enzymes of hepatic gluconeogenesis in periparturient dairy cows with induced fatty liver

The objective was to measure the activities of all the enzymes essential for hepatic gluconeogenesis in dairy cows with induced fatty liver. We aimed to induce severe fatty liver in ten experimental cows by overfeeding them during the dry period while seven control cows were maintained on a restricted diet. To induce a marked negative energy balance, the experimental cows were deprived of feed for 8 h immediately after parturition. In addition, the experimental cows were given a restricted amount of diet during the first 5 d of lactation. Liver samples were collected 1 week before and 1, 2 and 4 weeks after parturition. Before parturition, liver triacylglycerol concentrations did not differ between the two groups. After parturition, the experimental cows developed marked fatty liver as indicated by a higher level of triacylglycerols in the liver compared with the control cows. Before parturition, all gluconeogenic enzymes in the liver were lower in experimental cows than in control cows. Phosphoenolpyruvate carboxykinase, pyruvate carboxylase and propionyl-CoA carboxylase were significantly lower and fructose 1,6-bisphosphatase and glucose 6-phosphatase tended to be lower in the experimental cows. The activities of two crucial enzymes for gluconeogenesis in ruminants, i.e., phosphoenolpyruvate carboxykinase and propionyl-CoA carboxylase, remained low throughout the sampling period post partum. Activities of pyruvate carboxylase and glucose 6-phosphatase in the experimental cows post partum were upgraded to values similar to those of the control cows. The results showed that the capacity for hepatic gluconeogenesis before parturition was lower in cows with induced fatty liver than in control cows. After parturition, the low activities of crucial gluconeogenic enzymes indicated insufficient production of glucose. It is suggested that the low gluconeogenic capacity leads successively to low blood glucose concentrations, low insulin levels and high rates of mobilization of fatty acid, causing severe hepatic lipidosis.     

Technical note: Characterization of ceramide in bovine lipoproteins

The hepatic synthesis and export of ceramide is enhanced in diabetic monogastrics. Moreover, ceramide in lipoproteins can mediate the development of insulin resistance. We have previously demonstrated that circulating ceramide increases during the progression of insulin resistance in postpartum dairy cows. Considering that the origins of circulating ceramide required investigation, our objective was to develop a method to characterize the ceramide profile of lipoprotein fractions collected from dairy cows. Serum was collected from 4 nonpregnant and nonlactating Holstein dairy cows. Serum lipoproteins were isolated using size exclusion chromatography by fast protein liquid chromatography (SEC-FPLC). Measurement of triacylglycerol (TAG), phospholipid, total cholesterol, and protein was performed using standard colorimetry practices. Following lipid extraction, fractions were analyzed using electrospray ionization tandem mass spectrometry. Data were analyzed as repeated measures using a mixed model. Lipoprotein isolation using SEC-FPLC and subsequent colorimetric analyses confirmed the presence of 4 distinct fractions: TAG-rich, low density (LDL), and large (buoyant) and small (dense) high density lipoprotein (HDL) subclasses. As expected, the fraction representing mixed very low density lipoproteins and chylomicrons primarily contained TAG. Low density lipoprotein fractions were equally enriched with cholesterol and phospholipid. Buoyant HDL contained elevated levels of cholesterol, phospholipid, and protein. In contrast, the fraction containing dense HDL primarily contained protein. Our method revealed that LDL are enriched with ceramides. Ceramides were also compartmentalized to a lesser extent within both HDL subclasses and TAG-rich lipoproteins. Comparable to whole serum, C16:0-ceramide was the predominant ceramide quantified in all lipoprotein subclasses. Interestingly, the proportion of C24:0-ceramide to total ceramide was elevated in TAG-rich lipoproteins, relative to all other lipoprotein subclasses. We conclude that SEC-FPLC coupled with mass spectrometry is a means to quantify ceramides in lipoprotein fractions. Moreover, ceramides are enriched within bovine LDL, and lipoprotein ceramide profiles reflect levels observed in whole serum. Future investigation will need to determine the biological importance of lipoprotein ceramides with distinct C-chains at amide residues.     

Interrelation of fatty acid composition in adipose tissue, serum, and liver of dairy cows during the development of fatty liver postpartum

We investigated the composition of fatty acids in adipose tissue, serum, and liver of cows that were fed at restricted energy intake or were overfed during the dry period. Overfed cows had higher concentrations of serum nonesterified fatty acids and consequently accumulated greater amounts of triacylglycerols in the liver than did cows that were fed at restricted energy intake. The percentages of the different fatty acids present in adipose tissue were similar for both groups and did not change during sampling intervals. Before parturition, concentrations of the individual fatty acids present in serum were similar between groups. After parturition, concentrations of major fatty acids in serum, including palmitic, stearic, oleic, and linoleic acids significantly increased in both groups and were higher in overfed cows than in cows that were fed at restricted energy intake. The shift of concentrations of the different fatty acids present in the liver--as a result of increased lipolysis-was observed in palmitic, oleic, and linoleic acids but not stearic acid, suggesting that stearic acid is used by the liver (i.e., oxidation) or is considerably secreted through the milk, thereby not increasing in accumulation in the liver. In conclusion, different feeding regimens during the dry period do not influence the composition of fatty acids in adipose tissue. More intensive lipolysis results in increased concentrations of palmitic, stearic, oleic, and linoleic acids in the blood; subsequently, these fatty acids, excluding stearic acid, greatly accumulated in the liver.     

Effect of overfeeding during the dry period on the rate of esterification in adipose tissue of dairy cows during the periparturient period.

The in vitro rate of esterification of fatty acids in adipose tissue was compared between cows that were fed at restricted energy intake and cows that were overfed during the dry period. Subcutaneous adipose tissue was biopsied at -1, 0.5, 1, and 3 wk from parturition. The basal in vitro rate of esterification was quantified, as well as the rate of esterification after the addition of glucose or glucose plus insulin. The basal rate in adipose tissue from overfed cows at -1 wk was higher than in adipose tissue from cows that were fed at restricted energy intake and indicated enhanced storage of triacylglycerols in adipose tissue of overfed cows at that time. The rate of esterification after the addition of glucose or glucose plus insulin was increased in both groups at each sampling time, but the mean rates, expressed as a percentage of the basal rates, were lower for overfed cows than for cows that were fed at restricted energy intake at 0.5 and 1 wk. Although the addition of glucose or glucose plus insulin increased esterification rates in adipose tissue from both groups of cows, adipose tissue from overfed cows was less sensitive to the addition of these compounds. In conclusion, overfeeding during the dry period predisposed cows to accumulate fat in adipose tissue during the prepartum period. The smaller increase in esterification rate after the addition of glucose or glucose plus insulin in adipose tissue of overfed cows indicates a lower ability of the adipose tissue to esterify circulating fatty acids or to reesterify mobilized fatty acids, which, combined with higher rates of lipolysis postpartum, contributes to continuously elevated concentrations of circulating nonesterified fatty acids postpartum, leading to a more severe hepatic lipidosis in overfed cows.     

The effect of the transition period and postpartum body weight loss on macrophage infiltrates in bovine subcutaneous adipose tissue

The transition period in dairy cattle is characterized by a rapid increase in metabolic demands due to the onset of lactation in parallel with a voluntary drop in dry matter intake, which slowly increases compared with the rapidly increasing milk production. The resulting deficit in energy intake is largely met by lipolysis of adipose tissue stores liberating nonesterified fatty acids (NEFA) as energy substrates to support milk production and for the synthesis of milk fat. Previous work in cattle and other species has documented an increase in adipose tissue macrophages (ATM) during periods of feed restriction and lipolysis; however, alterations in ATM during the transition period have only recently received interest because of the role that adipose tissue remodeling and inflammation may play during this time. The primary objective of this study was to evaluate the number of macrophages in subcutaneous adipose tissue from dairy cattle at multiple time points throughout the transition period. Secondary objectives included the evaluation for potential associations of ATM with changes in body condition score (BCS) and concentrations of circulating NEFA and β-hydroxybutyrate. Holstein cows (n = 25) were enrolled in the study 28 d before expected parturition and fed the same prepartum and postpartum rations for ad libitum consumption. On d 10 before expected parturition, and on d 4 and 21 after calving, subcutaneous adipose tissue biopsies were taken from the paralumbar fossa and preserved in formalin. Paraffin-embedded sections of tissue from each of the 25 cows at the 3 time points (total of 75 samples) were labeled with an immunohistochemical marker of macrophages, ionized calcium-binding adapter molecule 1 (Iba1, also known as allograft inflammatory factor 1). The number of Iba1-positive cells, the number of crown-like structures, and the number of immunopositive aggregates were enumerated in each sample. The number of Iba1-positive macrophages increased by 60 and 93% on d 4 and 21, respectively, when compared with the average prepartum number. Additionally, cows that lost ≥0.50 points of BCS had a 57 and 52% higher number of ATM on d 4 and 21 postpartum, respectively, when compared with cows that lost ≤0.25 points of BCS. No association was found between ATM numbers and NEFA and β-hydroxybutyrate concentrations or milk production in early lactation. Cows experiencing a more pronounced decrease in BCS postpartum had the largest increase in adipose tissue macrophage counts, suggesting that a greater degree of adipose tissue remodeling during the period of greatest nutrient deficit may be associated with higher numbers of ATM.     

Prepartum dietary energy intake affects metabolism and health during the periparturient period in primiparous and multiparous Holstein cows

An experiment was conducted to determine the effect of prepartum plane of energy intake on metabolic profiles related to lipid metabolism and health in blood and liver. Primiparous (n = 24) and multiparous (n = 23) Holsteins were randomly assigned by expected date of parturition to 1 of 3 prepartum energy intakes. A high energy diet [1.62 Mcal of net energy for lactation (NE_L)/kg; 15% crude protein] was fed for either ad libitum intake or restricted intake to supply 150% (OVR) or 80% (RES) of energy requirements for dry cows in late gestation. To limit energy intake to 100% of National Research Council requirements at ad libitum intake, chopped wheat straw was included as 31.8% of dry matter for a control diet (CON; 1.21 Mcal of NE_L/kg of dry matter; 14.2% crude protein). Regardless of parity group, OVR cows had greater concentrations of glucose, insulin, and leptin in blood prepartum compared with either CON or RES cows; however, dietary effects did not carry over to the postpartum period. Prepartum nonesterified fatty acids (NEFA) were lower in OVR cows compared with either CON or RES cows. Postpartum, however, OVR cows had evidence of greater mobilization of triacylglycerol (TAG) from adipose tissue as NEFA were higher than in CON or RES cows, especially within the first 10 d postpartum. Prepartum β-hydroxybutyrate (BHBA) was not affected by diet before parturition; however, within the first 10 d postpartum, OVR cows had greater BHBA than CON or RES cows. Prepartum diet did not affect liver composition prepartum; however, OVR cows had greater total lipid and TAG concentrations and lower glycogen postpartum than CON or RES cows. Frequency of ketosis and displaced abomasum was greater for OVR cows compared with CON or RES cows postpartum. Controlling or restricting prepartum energy intake yielded metabolic results that were strikingly similar both prepartum and postpartum, independent of parity group. The use of a bulky diet controlled prepartum energy intake in multiparous and primiparous cows, improved metabolic status postpartum, and reduced the incidence of health problems. When metabolic profiles are considered collectively, cows overfed energy prepartum exhibited an “overnutrition syndrome” with characteristics of clinical symptoms displayed by diabetic or obese nonruminant subjects. This syndrome likely contributed to metabolic dysfunction postpartum.     

Unrestricted feed intake during the dry period impairs the postpartum oxidation and synthesis of fatty acids in the liver of dairy cows

The purpose of this study was to determine the activities of key hepatic enzymes of fatty acid synthesis and oxidation in cows that had excessive body fat at parturition. Dairy cows were allocated to either an experimental group or a control group. All cows were offered a total mixed ration with an energy content of 6.6 MJ of net energy for lactation per kilogram of dry matter and consisting of corn silage, beet pulp, rapeseed meal, and soybean meal. Control cows were restricted to 6.8 kg/dry matter of the mixed ration in the dry period. Experimental cows had unrestricted access to the mixed ration during the dry period to increase body fat and induce fatty liver postpartum. Blood and liver samples were collected 1 wk before and 1, 2, and 4 wk after parturition. Before parturition, neither the serum nonesterifled fatty acids nor the hepatic triacylglycerol concentrations differed between experimental and control cows. After parturition, the values for these variables were greater in experimental cows than in control cows. Plasma 3-hydroxybutyrate increased sharply after parturition in the experimental group. In liver, the activity of acetyl-CoA carboxylase was already significantly lower in the experimental group before parturition. After parturition, the activities of acetyl-CoA carboxylase and fatty acid synthase dropped in the experimental group. The activity of 3-hydroxy-acyl-CoA dehydrogenase in liver was less in experimental cows following parturition. Hepatic citrate synthase activity increased only in the control group after parturition. Unrestricted feed intake before parturition reduces de novo fatty acid synthesis as well as fatty acid oxidation after parturition. The reduction in fatty acid oxidation following parturition may contribute to postpartum accumulation of triacylglycerol in the livers of cows with unrestricted access to feed during the dry period.     

Feeding increasing amounts of ruminally protected choline decreased fatty liver in nonlactating,pregnant Holstein cows in negative energy status

The objectives were to determine the optimal feeding amount of choline in a ruminally protected form to reduce the triacylglycerol (TAG) concentration in liver and to increase TAG in blood plasma of dairy cows. Pregnant, nonlactating multiparous Holstein cows (n = 77) were blocked by body condition score (3.59 ± 0.33) and assigned to treatment at 64 ± 10 d before calculated calving date. Dietary treatments were top-dressing of 0, 30, 60, 90, or 120 g/d of ruminally protected choline (RPC; Balchem Corp., New Hampton, NY) ions to supply the equivalent of 0, 6.5, 12.9, 19.4, and 25.8 g/d of choline ions. Diets were formulated to exceed nutrient requirements for maintenance and pregnancy and fed in ad libitum amounts for the first 5 d. From d 6 to 15, cows were restricted to consume approximately 31% of their net energy requirements to simulate early lactating cows in negative energy balance. Methionine intake was maintained throughout each 15-d period. Liver was biopsied at 5 and 14 d and analyzed for TAG and glycogen. Blood was sampled on d 5 and 14 and plasma analyzed for glucose, insulin, cholesterol, β-hydroxybutyrate, long-chain fatty acids, and haptoglobin. On d 14, a mixture of saturated long-chain fatty acids, ground corn, and dried molasses (50:37:13) was offered (908 g, as-is basis) 10 h after the single daily feeding. Blood samples were collected for 19 h and plasma analyzed for TAG and cholesterol to assess apparent absorption of dietary fat. Mean dry matter intake and energy balance decreased from means of 9.5 to 3.3 kg/d and from 0.6 to ?9.2 Mcal of net energy for lactation/d during the ad libitum and restricted feeding periods, respectively. Plasma concentrations of the lipid-soluble choline biomolecules, namely total phosphatidylcholines, total lysophosphatidylcholines, and sphingomyelin, increased with choline supplementation. Feed restriction increased plasma concentrations of β-hydroxybutyrate and free long-chain fatty acids, whereas those of glucose, insulin, and total cholesterol decreased. During feed restriction, concentration of hepatic TAG and plasma haptoglobin decreased linearly, whereas concentration of hepatic glycogen tended to increase quadratically with increasing intake of RPC. After fat supplementation, mean plasma concentration of TAG increased by an average of 21% with intake of RPC ions, peaking at intakes of ≥6.5 g/d of RPC ion. In summary, feeding RPC ions to cows in negative energy balance had increasing lipotropic effects on the liver when consumed up to 25.8 g/d, whereas feeding only 6.5 g/d increased concentrations of hepatic glycogen and TAG in the blood.     

Hypolipidemic effect of proteoglycans isolated from sweet potato (Ipomoea batatas LAM) in hyperlipidemia rats

Effects of sweet potato proteoglycans (SPG) in rats fed high lipid diets were investigated. Serum lipid indices for SPG were evaluated. SPG had hypolipidemic effects that were correlated with dosage. There were no significant (p>0.05) differences in hypolipidemic effects between SPG varieties of different molecular weights. SPG-1 (Mw value: 508.3 kDa) significantly (p<0.05) reduced serum total cholesterol (TC), triacylglyceride (TAG), low density lipoprotein cholesterol (LDL-C), and apolipoprotein (Apo) B levels, and the arteriosclerosis index (AI), and induced serum high density lipoprotein cholesterol (HDL-C), ApoA-I, lecithin cholesterol acyltransferase (LCAT), and liver lipoprotein lipase (LPL) activities, compared with controls. The whole blood viscosity and red blood cell (RBC) membrane microviscosity were also decreased. Hypolipidemic effects were mediated via elevation of the serum HDL-C concentration by induction of LACT activity, an increased ApoA-I concentration, and a reduction in the serum lipid concentration due to regulation of LPL activity.     

The effects of two out-wintering pad systems compared with free-stalls on dairy cow hoof and limb health

Lameness is one of the most serious health and welfare problems for dairy cows. This study compared hoof health, limb health and locomotion of dairy cows in three over-winter management systems. Treatments were: (1) an indoor free-stall system (FS), (2) outdoors on an uncovered woodchip pad (UP) and (3) outdoors on a covered woodchip pad (CP). Animals were assigned to treatments at drying off, remained on treatment until parturition, then turned out to pasture. Sole lesions were scored on assignment to treatment, at calving, and 6 weeks and 12 weeks post partum. Locomotion and skin lesions were scored on assignment to treatment, and every 2 weeks until parturition. Post-partum locomotion was scored weekly for 13 weeks. More FS cows were affected by limb lesions than CP cows, and these animals had the worst locomotion scores post calving. There was an effect of inspection, and interactive effect of inspection and treatment on sole lesion scores. UP cows had the highest sole lesion scores 12 weeks post partum probably due to softening of the hooves arising from exposure to moisture while on the woodchip pad. However, post partum, UP cows had better locomotion scores, and CP cows tended to have better scores than cows in FS. Both of these treatments were less likely to have a high tracking-up score than cows in FS. It is possible that cows in this treatment were able to exercise more, owing to the secure underfoot surface and lack of barriers, and this promoted limb flexibility. Although woodchip pads provided benefits for limb health and locomotion, the wood chip surface in both outdoor systems did not offer protective benefits to the hooves relative to housing on concrete. However, the provision of shelter on the pad somewhat overcame the problem of high sole lesion scores in the post-partum period.     

Effects of prepartum 2,4-thiazolidinedione on insulin sensitivity, plasma concentrations of tumor necrosis factor-α and leptin, and adipose tissue gene expression

Administration of peroxisome proliferator-activated receptor gamma (PPARγ) ligands, thiazolidinediones (TZD), to prepartum dairy cattle has been shown to improve dry matter intake and decrease circulating nonesterified fatty acids (NEFA) around the time of calving. The objective of this work was to elucidate mechanisms of TZD action in transition dairy cattle by investigating changes in plasma leptin, tumor necrosis factor-α (TNFα), the revised quantitative insulin sensitivity check index (RQUICKI), and adipose tissue gene expression of leptin, PPARγ, lipoprotein lipase (LPL), and fatty acid synthase (FAS). Multiparous Holstein cows (n = 40) were administered 0, 2.0, or 4.0 mg of TZD/kg of body weight (BW) by intrajugular infusion once daily from 21 d before expected parturition until parturition. Plasma samples collected daily from 22 d before expected parturition through 21 d postpartum were analyzed for glucose, NEFA, and insulin. Plasma samples collected on d −14, −3, −1, 1, 3, 7, 14, and 49 relative to parturition were also analyzed for leptin and TNFα. Adipose tissue was collected on d 7 before expected parturition from a subset of cows, and gene expression was examined via quantitative real-time PCR. A tendency for a treatment by time effect on plasma leptin prepartum was observed such that values were similar on d −14 but cows receiving 2.0 mg/kg of BW of TZD tended to have lower circulating leptin as calving approached. Postpartum leptin tended to be increased linearly (2.3, 2.4, and 2.5 ± 0.1 ng/mL for 0, 2.0, and 4.0 mg/kg treatments, respectively) in cows that received TZD prepartum. Plasma TNFα increased linearly (2.6, 3.7, and 4.0 ± 0.1 pg/mL) in response to TZD treatment and decreased through the first week postpartum. Calculation of RQUICKI 1/[log(glucose) + log(insulin) + log(NEFA)] suggested altered insulin sensitivity in cows administered TZD that may depend on day relative to calving. Administration of TZD increased adipose tissue expression of PPARγ mRNA (11.0, 13.3, and 12.8 ± 1.9). Administration of TZD had a quadratic effect on gene expression of leptin (16.2, 10.7, and 17.4 ± 1.6) and no effect on LPL expression, and expression of FAS was lower for TZD-treated cows than for controls (8.2, 4.2, and 6.1 ± 1.8, respectively). Results imply altered expression and plasma concentrations of leptin, increased plasma TNFα concentrations, and increased expression of PPARγ in adipose tissue as potential mechanisms for the effects of TZD administration on transition dairy cattle.     

Effect of duration of supplementation of selenium and vitamin E on periparturient dairy cows

Cows were fed diets either supplemented with .2 ppm Se and 70 IU vitamin E/kg diet DM (21 cows) or unsupplemented (40 cows) during the dry period (approximately 60 d). From parturition to 21 d of lactation, cows were fed diets that were either supplemented with .3 ppm Se and 40 IU/kg vitamin E or unsupplemented. At d 21 following parturition, 18 cows fed the unsupplemented diet were switched to diets containing 0 or .3 ppm supplemental Se and 0 or 40 IU/kg supplemental vitamin E arranged factorially. These diets were fed for the next 32 d. The remaining cows continued their respective diets for 32 d. Plasma Se concentrations averaged .1 microgram/ml for supplemented cows but were .05 micrograms/ml for unsupplemented cows. Plasma Se concentration from cows fed supplemental Se from 21 to 53 d postpartum increased rapidly and were not different from long-term supplemented cows. Whole blood glutathione peroxidase activity was lower in unsupplemented than in supplemented cows. Short-term Se supplementation increased glutathione peroxidase activity above that for unsupplemented animals, but activity was still less than that in long-term supplemented animals. Plasma alpha-tocopherol concentrations at parturition and d 21 postpartum were lower in unsupplemented than in supplemented animals. On d 53 postpartum, no differences in plasma alpha-tocopherol concentrations were found between long-term supplemented and unsupplemented cows. Supplementing vitamin E during the dry period increased alpha-tocopherol content of colostrum.     

Limit-feeding a high-energy diet to meet energy requirements in the dry period alters plasma metabolite concentrations but does not affect intake or milk production in early lactation

Limit-feeding dry cows a high-energy diet may enable adequate energy intake to be sustained as parturition approaches, thus reducing the extent of negative energy balance after parturition. Our objective was to evaluate the effect of dry period feeding strategy on plasma concentrations of hormones and metabolites that reflect energy status. Multiparous Holstein cows (n = 18) were dried off 45 d before expected parturition, paired by expected calving date, parity, and previous lactation milk yield, and randomly assigned to 1 of 2 dry-period diets formulated to meet nutrient requirements at ad libitum or limited intakes. All cows were fed the same diet for ad libitum intake after parturition. Prepartum dry matter intake (DMI) for limit-fed cows was 9.4 kg/d vs. 13.7 kg/d for cows fed ad libitum. During the dry period, limit-fed cows consumed enough feed to meet calculated energy requirements, and ad libitum-fed cows were in positive calculated net energy for lactation (NE_L) balance (0.02 vs. 6.37 Mcal/d, respectively). After parturition, milk yield, milk protein concentration, DMI, body condition score, and body weight were not affected by the prepartum treatments. Cows limit fed during the dry period had a less-negative calculated energy balance during wk 1 postpartum. Milk fat concentration and yield were greater for the ad libitum treatment during wk 1 but were lower in wk 2 and 3 postpartum. Plasma insulin and glucose concentrations decreased after calving. Plasma insulin concentration was greater in ad libitum-fed cows on d −2 relative to calving, but did not differ by dietary treatment at other times. Plasma glucose concentrations were lower before and after parturition for cows limit-fed during the dry period. Plasma nonesterified fatty acid concentrations peaked after parturition on d 1 and 4 for the limit-fed and ad libitum treatments, respectively, and were greater for limit-fed cows on d −18, −9, −5, and −2. Plasma tumor necrosis factor-α concentrations did not differ by treatment in either the pre- or postpartum period, but tended to decrease after parturition. Apart from a reduction in body energy loss in the first week after calving, limit feeding a higher NE_L diet during the dry period had little effect on intake and milk production during the first month of lactation.     

Effects of prepartum roughage neutral detergent fiber levels on periparturient dry matter intake, metabolism, and lactation in heat-stressed dairy cows

Heat stress of lactating cattle results in dramatic reductions in dry matter intake (DMI). As a result, energy input cannot satisfy energy needs and thus accelerates body fat mobilization. Decreasing the level of roughage neutral detergent fiber (NDF) in prepartum diets, and thereby increasing the amount of nonfiber carbohydrates, may provide an adequate supply of energy and glucose precursors to maintain and minimize the decrease in DMI while reducing mobilization of adipose tissue. The effects of 3-wk prepartum diets containing different amounts of roughage NDF on DMI, blood metabolites, and lactation performance of dairy cows were investigated under summer conditions in Thailand. Thirty cross-bred cows (87.5% Holstein × 12.5% Sahiwal) were dried off 60 d before their expected calving date and were assigned immediately to a nonlactating cow diet containing the net energy for lactation recommended by the National Research Council (2001) model. The treatment diets contained 17.4, 19.2, and 21.0% DM as roughage NDF from bana grass (Pennisetum purpureum × Pennisetum glaucum) silage. Levels of concentrate NDF were 39.8, 40.2, and 38.6% of dietary NDF, so the levels of dietary NDF were 28.9, 32.1, and 34.2% of DM. After parturition, all cows received a lactating cow diet containing 12.7% roughage NDF and 23% dietary NDF. During the entire experiment, the minimum and maximum temperature-humidity index averaged 77.7 and 86.8, respectively, indicating conditions appropriate for the induction of extreme heat stress. As parturition approached, DMI decreased steadily, resulting in a 12.9, 25, and 32.8% decrease in DMI from d −21 until calving for nonlactating cows fed prepartum diets containing 17.4, 19.2, and 21% roughage NDF, respectively. During the 3-wk prepartum period, intakes of DM and net energy for lactation and concentrations of plasma glucose and serum insulin were higher for cows fed diets containing less roughage NDF. In cows fed the 3-wk prepartum diets containing less roughage NDF, calf birth weights, milk yield, and 4% fat-corrected milk were higher, whereas periparturient concentrations of serum nonesterified fatty acids and plasma β-hydroxybutyrate were lower. There was a carryover effect of the prepartum diet on serum nonesterified fatty acids and plasma β-hydroxybutyrate during the first 7 d in milk, and therefore on milk production. These results suggest that feeding diets containing decreased amounts of roughage NDF during the 3-wk prepartum period may minimize the decrease in DMI and lipid mobilization as parturition approaches. This strategy may thus minimize the effect of hormonal factors and heat stress on periparturient cows.     

Metabolism and lactation performance in dairy cows fed a diet containing rumen-protected fat during the last twelve weeks of gestation

Effects of dietary fat supplementation prepartum on liver lipids and metabolism in dairy cows are contradictory. Thus, we examined in 18 German Holstein cows (half-sib; first lactation 305-d milk yield >9,000 kg) whether dietary fat:carbohydrate ratio during the last trimester of gestation affects lipid metabolism and milk yield. The diets were formulated to be isoenergetic and isonitrogenous but differed in rumen-protected fat (FD; 28 and 46.5 g/kg of dry matter during far-off and close-up dry period; mainly C16:0 and C18:1) and starch concentration [carbohydrate diet (CD); 2.3 times as much starch as FD]. Diets were given ad libitum starting 12 wk before expected parturition. After parturition all cows were fed a single lactation diet ad libitum for 14 wk. With the FD treatment, dry matter intake was depressed prepartum, milk yield during first 4 wk of lactation was lower (36.9 vs. 41.0 kg/d), and postpartum energy balance during this period was more negative. During the first 4 wk, cows in the FD group had lower lactose percentage and yield but higher milk fat, whereas milk protein and fat yield as well as energy-corrected milk did not differ. Between wk 5 and 14, milk fat and milk protein percentage was lower in CD than in FD. Milk fat C14:0 was lower and C16:1 was higher in the FD group. For FD cows, plasma triacylglycerol, nonesterified fatty acids, and cholesterol concentrations were higher prepartum, whereas plasma β-hydroxybutyrate and glucose concentrations were lower. During the first 10 d after parturition, plasma triacylglycerol concentration was higher in FD, and prepartum plasma glucose and cholesterol differences persisted during the first 14 wk of lactation. Irrespective of prepartum nutrient composition, concentrations of plasma leptin and subcutaneous fat leptin mRNA decreased between −10 d to +10 d relative to parturition, and liver lipids and glycogen reached maximum and minimal values, respectively, 10 d after parturition. Acetyl-coenzyme A carboxylase α mRNA abundance in subcutaneous fat decreased between −10 d to +1 d relative to parturition by 97%, whereas it was generally much lower in the liver and remained at a low level until wk 14 of lactation. In conclusion, feeding a diet containing rumen-protected fat during late lactation and dry period until calving negatively affected dry matter intake, energy balance, and milk yield during subsequent lactation, did not change acetyl-coenzyme A carboxylase α mRNA abundance in subcutaneous fat, and was not beneficial for liver lipid accumulation.     

Effect of various dietary fats and proteins on serum cholesterol level in rats

A 60 day experiment on growing rats was performed to evaluate the effect of two protein sources (casein and soybean protein isolate) and four fats (olive oil, OO; evening primrose oil, EO; linseed oil, LO; and butter, B) on plasma lipoprotein fractions: total cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C), low‐density lipoprotein cholesterol (LDL‐C) and triacylglycerol (TAG) levels. The different protein and fat sources had no influence on body weight gain of rats. Fats had no significant effect on TC and LDL‐C. Significant differences were found only in HDL‐C. OO gave the lowest (43.3 mg dl^?1) and EO the highest (54.4 mg dl^?1) value. Proteins had a significant influence on all lipids analysed. Soybean protein caused higher TC (78.5 vs 67.9 mg dl^?1), HDL‐C and TAG levels but significantly lowered LDL‐C in the plasma of rats. These results suggest that protein has a stronger effect on cholesterol level than fat. © 2002 Society of Chemical Industry     

Prepartal plane of nutrition, regardless of dietary energy source, affects periparturient metabolism and dry matter intake in Holstein cows

Previous research in our laboratory showed that dietary fat supplementation during the dry period was associated with decreased peripartum hepatic lipid accumulation. However, fat supplementation decreased dry matter (DM) intake and thereby confounded results. Consequently, 47 Holstein cows with body condition scores (BCS) ≤ 3.5 at dry-off were used to determine whether source or amount of energy fed to dry cows was responsible for the decreased hepatic lipid content. Moderate grain- or fat-supplemented diets [1.50 Mcal of net energy for lactation (NE_L)/kg] were fed from dry-off (60 d before expected parturition) to calving at either ad libitum (160% of NE_L requirement) or restricted (80% of NE_L requirement) intakes. Postpartum, cows were fed a single lactation diet for ad libitum intake and performance was measured for 105 d. Prepartum intakes of DM and NE_L were significantly lower for feed-restricted cows as designed. During the first 21 d postpartum, previously restricted cows had higher intakes of DM and NE_L. Body weights and BCS were lower prepartum for restricted cows but groups converged to similar nadirs postpartum. Restricted-fed cows had lower concentrations of glucose and insulin and increased concentrations of NEFA in plasma during the dry period. Peripartum NEFA rose markedly for all treatments but were higher postpartum for cows previously fed ad libitum. Plasma concentrations of NEFA and BHBA remained lower in cows restricted-during the dry period. Postpartum concentrations of total lipid and triglyceride in liver were lower in cows previously feed-restricted. Across dietary treatments, activity of carnitine palmitoyltransferase (CPT) in hepatic mitochondria was lowest at − 21 d, highest at 1 d, and decreased at 21 and 65 d relative to parturition. The activity of CPT at d 1 tended to be higher for previously feed-restricted cows; thereafter, CPT activity declined more rapidly than in cows fed ad libitum. Nutrient intake during the dry period had more pronounced effects on peripartal lipid metabolism and DMI than did composition of the prepartum diet.     

Effect of vitamin E supplementation on neutrophil function, milk composition and plasmin activity in dairy cows in a commercial herd

Fifty-six Holstein dairy cows from a commercial dairy herd in the Northern part of Greece were used to determine the effect of vitamin E supplementation on immune parameters, milk composition and milk quality. Cows were assigned to one of two experimental groups: control (no vitamin E supplementation) and vitamin E supplementation. Supplementation of vitamin E started 4 weeks prior to and continued up to 12 weeks after parturition. Supplementation included daily oral administration of vitamin E at 3000 i.u./cow prepartum and was reduced to 1000 i.u./cow post partum. Blood samples were collected weekly for 8 weeks starting 4 weeks before parturition, neutrophils were isolated and the following parameters were determined in neutrophils activated by phorbol myristate acetate: total cell-associated and membrane-bound urokinase plasminogen activator (u-PA) activity and superoxide production. Milk samples were collected weekly and fat, protein, lactose, somatic cell count (SCC), plasmin and plasminogen-derived activity were determined. Activated neutrophils isolated from cows that received supplemental vitamin E had higher (P<0.01) total and membrane-bound u-PA activities during the first 3 weeks after parturition and higher (P<0.01) superoxide production during week 1 prepartum and week 1 post partum compared with the corresponding values of activated neutrophils isolated from control cows. Vitamin E supplementation had no effect (P=0.28) on plasminogen-derived activity in milk. Milk obtained from cows that received supplemental vitamin E had SCC lower by 25% (P<0.05) and plasmin lower by 30% (P<0.01) than corresponding values in milk obtained from control cows. The reduction in plasmin as a result of vitamin E supplementation is very beneficial to the dairy industry because plasmin reduces the cheese-yielding capacity of milk, affects the coagulating properties of milk and its overall ability to withstand processing during cheesemaking. In conclusion, vitamin E supplementation had positive effects on the function of bovine neutrophils and milk quality in a commercial dairy herd.     

Lipid metabolite profiles and milk production for Holstein and Jersey cows fed rumen-protected choline during the periparturient period

Choline is important for assembly of very low density lipoproteins to export triglyceride from liver; however, studies to assess the effect of rumen-protected choline (RPC) supplementation on blood lipid metabolites in periparturient dairy cows have not been conducted. Thirty-two multiparous Holstein and 10 multiparous Jersey cows were randomly assigned to control or RPC treatments. A close-up diet was fed from approximately 3 wk before parturition through parturition, followed by a lactation diet from parturition through 49 d postpartum. For RPC, diets were top-dressed once daily with 60 g of a RPC product (25% choline as choline chloride) from 21 d before expected parturition through 21 d postpartum. Treatment did not affect dry matter intake either prepartum (12.0 vs. 12.1 kg/d for RPC and control, respectively) or during the first 3 wk postpartum (14.8 vs. 15.7 kg/d, respectively). Daily yields of 3.5% fat-corrected milk (39.4 vs. 37.4 kg/d), fat (1.46 vs. 1.38 kg/d), and protein (1.09 vs. 1.05 kg/d) did not differ statistically by treatment (RPC vs. control, respectively). Jersey cows in the control group had lower concentrations of nonesterified fatty acids and β-hydroxybutyrate in plasma during d 1 to 10 postpartum than did other breed and treatment combinations. Cows fed RPC tended to have greater serum triglycerides prepartum (17.0 vs. 14.7 mg/dL) and lower plasma phospholipid at parturition (65.2 vs. 78.1 mg/dL) than control cows. Treatment did not affect cholesterol and phospholipid at other time points, but concentrations followed patterns of dry matter intake pre- and postpartum. Cows were in moderate body condition score (mean = 3.3) at the start of the study and did not lose excessive condition by 3 wk postpartum (mean body condition score loss = 0.5); therefore, cows might not have been at great risk for hepatic lipid accumulation. Additionally, calculated Met balance was negative postpartum; supplemental RPC might not have spared enough Met to produce a physiological benefit. More research is needed to determine how choline affects prevention or alleviation of fatty liver syndrome and to confirm potential differences between Holstein and Jersey cows.