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.     

Metabolic effects of abomasal L-carnitine infusion and feed restriction in lactating Holstein cows

l-Carnitine is required for mitochondrial fatty acid oxidation, but the effects of carnitine supplementation on nutrient metabolism during dry matter intake depression have not been determined in dairy cows. Studies in other species have revealed responses to l-carnitine that may be of specific benefit to dairy cows during the periparturient period. Eight lactating Holstein cows (132 ± 36 d in milk) were used in a replicated 4 × 4 Latin square experiment with 14-d periods. Treatments were factorial combinations of abomasal infusion of either water or l-carnitine (20 g/d; d 5 to 14) and either ad libitum or restricted intake (50% of previous 5-d dry matter intake; d 10 to 14) of a balanced lactation diet. Liver and muscle biopsies were obtained on d 14 of each period. Feed restriction induced negative balances of energy and metabolizable protein. In feed-restricted cows, carnitine infusion increased 3.5% fat-corrected milk yield compared with those infused with water. Total carnitine concentration in liver was increased in feed-restricted cows infused with carnitine but not in feed-restricted cows infused with water. Carnitine infusion stimulated in vitro oxidation of [1-¹⁴C] palmitate to acid-soluble products and decreased the proportion of [1-¹⁴C] palmitate that was converted to esterified products by liver slices. Feed-restricted cows infused with carnitine had lower liver total lipid concentration and tended to have decreased triglyceride accumulation compared with feed-restricted cows infused with water. Plasma nonesterified fatty acid concentration was not altered by carnitine infusion but was increased by feed restriction; serum β-hydroxybutyric acid was increased by carnitine infusion in feed-restricted cows. In cows fed for ad libitum intake, carnitine infusion affected β-hydroxybutyric acid, insulin, and urea N in serum, liver glycogen concentration, and in vitro alanine oxidation by liver slices, suggesting that hepatic and peripheral nutrient metabolism was influenced. l-Carnitine infusion effectively decreased liver lipid accumulation during feed restriction as a result of greater capacity for hepatic fatty acid oxidation. Further research examining dietary supplementation of l-carnitine during the periparturient period is warranted.     

Effect of feed restriction on reproductive and metabolic hormones in dairy cows

The objective of this trial was to evaluate the effects of feed restriction (FR) on serum glucose, nonesterified fatty acids, progesterone (P4), insulin, and milk production in dairy cows. Eight multiparous Holstein cows, 114 ± 14 d pregnant and 685 ± 39 kg of body weight, were randomly assigned to a replicated 4 × 4 Latin square design with 14-d periods. During the first 8 d of each period, cows in all treatments were fed for ad libitum feed intake. Beginning on d 9 of each period, cows received 1 of 4 treatments: ad libitum (AL), 25% feed restriction (25FR), 50% feed restriction (50FR), and 50% of TMR replaced with wheat straw (50ST). Daily feed allowance was divided into 3 equal portions allocated every 8 h with jugular blood samples collected immediately before each feeding through d 14. In addition, on d 12 of each period, blood samples were collected before and at 60, 120, 180, 240, 300, 360, 420, and 480 min after morning feeding. The conventional total mixed ration and total mixed ration with straw averaged 15.1 and 10.8%, 32.1 and 50.5%, and 26.8 and 17.0% for concentrations of crude protein, neutral detergent fiber, and starch, respectively. Cows that were feed and energy restricted had reduced dry matter intake, net energy for lactation intake, circulating glucose concentrations, and milk production, but greater body weight and body condition score losses than AL cows. Circulating concentrations of insulin were lower for cows fed 50FR (8.27 μIU/mL) and 50ST (6.24 μIU/mL) compared with cows fed AL (16.65 μIU/mL) and 25FR (11.16 μIU/mL). Furthermore, the greatest plasma nonesterified fatty acids concentration was observed for 50ST (647.7 μEq/L), followed by 50FR (357.5 μEq/L), 25FR (225.3 μEq/L), and AL (156.3 μEq/L). In addition, serum P4 concentration was lower for cows fed AL than cows fed 50ST and 25FR. Thus, FR reduced circulating glucose and insulin but increased P4 concentration, changes that may be positive in reproductive management programs.     

Effects of garlic and juniper berry essential oils on ruminal fermentation and on the site and extent of digestion in lactating cows

The objective of this study was to evaluate the effects of feeding essential oils from garlic (GAR) and juniper berry (JUN), or monensin (MO) on feed intake, ruminal fermentation, the site and extent of digestion, microbial protein synthesis, milk production, and immune status in dairy cows. Four midlactating Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 × 4 Latin square design with 21-d periods and 4 treatments: control (no additive), MO (330 mg/cow per d), GAR (5 g/cow per d), and JUN (2 g/cow per d). Cows were fed ad libitum a TMR consisting of 40% forage and 60% barley-based concentrate. Dry matter intake averaged 20.4 kg/d and was not affected by dietary additives. Total tract digestibilities of dry matter, organic matter, fiber, and starch were not affected by experimental treatments. However, ruminal digestibilities of dry matter and organic matter were higher (+13%) for GAR and JUN than for the control diet, mainly because of increased crude protein digestion in the rumen. Feeding GAR and JUN increased ruminal digestion of dietary protein by 11% as compared with the control. In contrast, ruminal digestion of dietary protein was reduced by 11% with MO as compared with the control. Milk fat content was lower for MO (2.68%) than for the GAR (3.46%), JUN (3.40%), and control (3.14%) diets. No effects of GAR, JUN, or MO were observed on milk production, ruminal microbial protein synthesis, ruminal pH, and ruminal concentrations of volatile fatty acids and ammonia N. The total and differential numbers of white blood cells as well as serum amyloid A and haptoglobin were not affected by the treatments, suggesting that additives had no effect on the immune status of cows. Results of this study indicate that supplementing dairy cows with GAR (5 g/d) and JUN (2 g/d) essential oils improved feed digestibility in the rumen, but possibly at the expense of a reduction in the flow of bypass protein to the small intestine. Feeding monensin could be beneficial in terms of increasing bypass protein from the rumen but did not improve feed digestion or milk production under the current experimental conditions.     

Evaluation of portal blood flow using transcutaneous and intraoperative Doppler ultrasonography in dairy cows with fatty liver

The objective of the study was to investigate portal blood flow (PBF) in dairy cows with fatty liver by means of Doppler ultrasonography. Eighty lactating German Holstein cows less than 100 d in milk were used (mean ± standard error of the mean; body weight: 583 ± 9 kg, age: 5 ± 0.2 yr, withers height: 145.4 ± 0.5 cm, milk yield: 9 ± 0.6 kg). All cows had left abomasal displacement and underwent omentopexy via right flank laparotomy. The size of the liver and the thickness over the portal vein were determined ultrasonographically. Doppler ultrasonographic examinations of PBF were carried out transcutaneously and intraoperatively directly via liver surface. The PBF velocities [peak maximum (v_max), peak minimum (v_min), and mean maximum (v_mean) velocity] were recorded. Venous pulsatility index (VPI) was calculated. Because transcutaneous Doppler ultrasonography revealed images of very poor quality in 58 of the 80 cows, only data obtained intraoperatively were presented. Liver biopsies were used for hepatic triacylglycerol (TAG) determination and histological examination. Based on histopathologic and ultrasonographic examinations, none of the cows suffered from hepatic disorders other than hepatic lipidosis. Hepatic TAG content ranged from 5 to 292 mg/g of liver fresh weight (FW). Cows were allocated to 1 of 4 groups according to their hepatic TAG content (very severe: TAG >150 mg/g of FW, n = 27; severe: >100-150 mg/g of FW, n = 18; moderate: ≥50-100 mg/g of FW, n = 19; mild: <50 mg/g of FW, n = 16). The VPI decreased with increasing TAG content (r = −0.55). The VPI did not differ between cows with severe and very severe fatty liver but it differed between cows of these 2 groups and cows with mild and moderate fatty liver. Velocities of PBF (v_mean, v_min, v_max) correlated negatively with hepatic TAG content (r = −0.26 to −0.37). Mean PBF velocity of the cows with very severe fatty liver differed from cows with severe, moderate, and mild fatty liver. Variables of PBF were inversely related to hepatic size and thickness (r = −0.06 to −0.35). In conclusion, the lower VPI and PBF velocities in cows with fatty liver and the negative correlations with the degree of hepatosteatosis may be explained by a reduction of vascular compliance in the liver because of fatty infiltration. These changes, which are believed to result from parenchymal swelling, were particularly pronounced when hepatic TAG content exceeded 150 mg/g FW.     

The effect of body condition score at calving and supplementation with Saccharomyces cerevisiae on milk production, metabolic status, and rumen fermentation of dairy cows in early lactation

The objective of this study was to examine the effects of live yeast (LY) supplementation and body condition score (BCS, 1-5 scale) at calving on milk production, metabolic status, and rumen physiology of postpartum (PP) dairy cows. Forty Holstein-Friesian dairy cows were randomly allocated to a 2 × 2 factorial design and blocked by yield, parity, BCS, and predicted calving date. Treatments were body condition at calving (low for BCS ≤3.5 or high for BCS ≥3.75; n = 20) and supplementation with LY (2.5 and 10 g of LY/d per cow for pre- and postcalving, respectively; control, no LY supplementation; n = 20). The supplement contained 10⁹ cfu of Saccharomyces cerevisiae/g (Yea-Sacc¹⁰²⁶ TS, Alltech Inc., Nashville, TN). Daily milk yield, dry matter intake, milk composition, BCS, body weight, and backfat thickness were recorded. Blood samples were harvested for metabolite analysis on d 1, 5, 15, 25, and 35 PP. Liver samples were harvested by biopsy for triacylglycerol (TAG) and glycogen analysis on d 7 precalving, and on d 7 and 21 PP. Rumen fluid was sampled by rumenocentesis for all cows on d 7 and 21 PP. Supplementation with LY had no effect on milk yield, dry matter intake, rumen fluid pH, or blood metabolites concentration of dairy cows with high or low BCS at calving. Feeding LY increased rumen acetate proportion and protozoal population, tended to increase liver glycogen, and decreased rumen ammonia nitrogen during early lactation. Over-conditioned cows at calving had greater body reserve mobilization and milk production and lower feed intake, whereas cows with a moderate BCS at calving had greater feed intake, lower concentrations of nonesterified fatty acids and β-hydroxybutyrate, lower liver TAG and TAG:glycogen ratio, and faster recovery from body condition loss. Additionally, the data suggest that concentrations of liver enzymes in blood might be used as an indicator for liver TAG:glycogen ratio. Results indicate that in the case of this experiment, where the control treatment was associated with an acceptable rumen pH, feeding yeast did not significantly improve indicators of energy status in dairy cows.     

Effects of dairy cow diet forage proportion on duodenal nutrient supply and urinary purine derivative excretion

Four mature Holstein-Friesian dairy cows were used in a 4 × 4 Latin square change-over design experiment made up of four 4-wk periods to investigate the relationship between microbial protein flow to the duodenum and excretion of purine derivatives (PD) in the urine. Four dietary treatments based on ad libitum access to ryegrass silage were offered, with a standard dairy concentrate included at different forage:concentrate (F:C) ratios, calculated on a dry matter basis: 80:20, 65:35, 50:50, and 35:65. Feed intakes increased as the proportion of concentrate in the diet increased, despite a concurrent decrease in silage intake. Increased feed intake led to increased nutrient flow to the duodenum. Milk yields increased as the diet F:C ratio decreased, with cows offered the 35:65 diet yielding nearly 8 kg/d more milk than cows offered the 80:20 diet; the concentrations of milk fat decreased and milk protein increased with a decreasing F:C ratio. Purine derivative excretion in the urine increased with an increasing proportion of concentrate in the diet, and there was a strong linear relationship between total PD excretion (allantoin and uric acid) and microbial N flow to the duodenum: microbial N (g/d) = 19.9 + 0.689 × total PD (mmol/d); R = 0.887. This strengthens the case for using PD excretion as a noninvasive marker of microbial protein flow from the rumen in dairy cows.     

Alfalfa cut at sundown and harvested as baleage improves milk yield of late-lactation dairy cows

Alfalfa (Medicago sativa L.) cut at sundown has been shown to contain greater concentration of total nonstructural carbohydrates (TNC) than that cut at sunup. Fourteen multiparous (8 ruminally cannulated) and 2 primiparous lactating dairy cows were randomly assigned to 2 treatments in a crossover design (2 periods of 24 d) to investigate the effects of alfalfa daytime cutting management on ruminal metabolism, nutrient digestibility, N balance, and milk yield. Half of each alfalfa field (total of 3 fields) was cut at sundown (PM) after a sunny day, whereas the second half was cut at sunup (AM) on the following day. Both PM and AM cuts were field-wilted and harvested as baleage (531 ± 15.0 g of dry matter/kg of fresh matter). Bales (PM and AM) were ranked according to their concentrations of TNC, paired, and each pair of PM and AM baleages was then assigned to each experimental day (total of 48 d). The difference in TNC concentration between PM and AM baleages fed during the 10 d of data and sample collection varied from −10 to 50 g/kg of dry matter. Each pair of baleage was fed ad libitum to cows once daily with no concentrate. Ruminal molar proportion of acetate and total volatile fatty acid concentration were greater in animals fed the AM baleage, whereas the proportion of valerate was greater with PM baleage; no other significant changes in ruminal molar proportions of volatile fatty acids were observed between forage treatments. Digestible organic matter intake, organic matter digestibility, and plasma Lys concentration were significantly greater in cows fed PM alfalfa, suggesting that more nutrients were available for milk synthesis. Significantly lower body weight gain and retained N as a proportion of N intake were observed in cows fed PM alfalfa, thus suggesting that nutrients were channeled to milk synthesis rather than to body reserves. Intake of dry matter (+1.0 kg/d), and yields of milk (+1.0 kg/d), milk fat (+70 g/d), and milk protein (+40 g/d) were significantly greater in cows fed PM vs. AM alfalfa. Concentration of milk urea N and excretion of urea N as a proportion of total urinary N were significantly reduced, and milk N efficiency was increased when feeding PM vs. AM alfalfa, indicating an improvement in N utilization. Increasing the TNC concentration of alfalfa by shifting forage cutting from sunup to sundown improved N utilization and milk production in late-lactation dairy cows.     

Effects of rumen acid load from feed and forage particle size on ruminal pH and dry matter intake in the lactating dairy cow

The objective of this study was to evaluate the effects of level of concentrate acidogenic value (AV) and forage particle size on ruminal pH and feed intake in lactating dairy cows. Two isoenergetic (net energy for lactation = 1.5 ± 0.01 Mcal/kg) and isonitrogenous (crude protein = 17.4 ± 0.1% dry matter) concentrates with either a low AV or high AV were formulated and fed in a total mixed ration with either coarsely or finely chopped corn silage and alfalfa haylage ad libitum. Four rumen-fistulated cows (114 ± 14 d in milk) were randomly assigned to 1 of the 4 treatments in a 4 × 4 Latin square with a 2 × 2 factorial treatment arrangement. Each period consisted of 3-wk (14-d treatment adaptation and 7-d data collection). Increasing the concentrate AV decreased the mean pH (from 6.07 to 5.97) and minimum pH (from 5.49 to 5.34). Cows fed high-AV diets spent a longer time below pH 5.6 (135.1 vs. 236.7 min/d; low-AV diet vs. high-AV diet, respectively) and pH 5.8 (290.0 vs. 480.6 min/d; low-AV diet vs. high-AV diet, respectively) than cows fed low-AV diets. Increasing forage particle size had no effect on the mean and minimum ruminal pH. There was an interaction between concentrate AV and forage particle size on maximum ruminal pH. Increasing forage particle size increased the maximum pH for cows fed the high-AV concentrate (6.69 vs. 6.72; low-AV diet vs. high-AV diet, respectively) and had no effect on the maximum pH for cows fed the low-AV concentrate (6.98 vs. 6.76; low-AV diet vs. high-AV diet, respectively). Increasing the concentrate AV did not affect dry matter intake but reduced neutral detergent fiber intake from 9.7 to 8.8 kg/d. Milk fat content was negatively correlated with time and area below pH 5.6 (time below, r = −0.51; area below, r = −0.56) and pH 5.8 (time below, r = −0.42; area below, r = −0.54). These results suggest that coarse forage particle size can attenuate drops in ruminal pH. However, the ameliorating effects of forage particle size on drops in ruminal pH were more apparent for high-AV diets than for low-AV diets. The AV approach combined with physically effective neutral detergent fiber would therefore improve the formulation of diets and help to mitigate subacute ruminal acidosis in dairy cows.     

Intake and performance of lactating cows grazing diverse forage mixtures

Twenty multiparous Holstein cows in midlactation grazed pastures of 4 forage mixtures in a 12-wk study repeated during 2 grazing seasons to determine if forage mixture complexity affected intake and productivity of lactating dairy cows. The forage mixtures were 1) orchardgrass plus white clover [2 species (SP)]; 2) orchardgrass, white clover, and chicory (3SP); 3) orchardgrass, tall fescue, perennial ryegrass, red clover, birdsfoot trefoil, and chicory (6SP); and 4) 6SP mixture plus white clover, alfalfa, and Kentucky bluegrass (9SP). Total herbage intake was similar among forage mixtures, averaging 12.0 kg/d across all forage mixtures and years. Milk production and composition were not affected by forage mixture or year, and averaged 34.6 kg/d, 3.4%, and 2.8% for milk production, milk fat percentage, and milk protein percentage, respectively. The conjugated linoleic acid content of milk fat was higher for cows that grazed the 3SP, 6SP, and 9SP mixtures than from cows that grazed the 2SP mixture (1.02 vs. 0.87 g of conjugated linoleic acid/100 g of fatty acids, respectively). Blood glucose, blood urea nitrogen, and nonesterified fatty acids were not affected by forage mixture and averaged 69.2 mg/dL, 13.4 mg/dL, and 277.5 μEq/L, respectively. The results of this study indicate that altering the forage mixture in pastures did not affect dry matter intake, milk production, or blood metabolite profiles of lactating cows. The use of complex mixtures of forages in grazing systems should not affect dairy cow performance.     

Effects of feed delivery time on feed intake, milk production, and blood metabolites of dairy cows

The objective of the study was to determine the effects of feed delivery time and its interactions with dietary concentrate inclusion and parity on milk production and on 24-h averages and patterns of feed intake and blood metabolites. Four multiparous and 4 primiparous lactating Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Experimental periods included 14 d of adaptation and 7 d of sampling. A higher concentrate diet with a forage:concentrate ratio (dry matter basis) of 38:62 or a lower-concentrate diet with a forage:concentrate ratio of 51:49 was delivered at either 0900 or 2100 h. During sampling periods, daily feed intakes, as well as feed intakes during 3-h intervals relative to feed delivery, were determined. During 2 nonconsecutive days of the sampling period, jugular blood was sampled every 2 h. Average temperature and relative humidity in the experimental facility were 20.4°C and 68.1%, and the maximum daily air temperature did not exceed 25°C. This data does not suggest that cows were heat-stressed. Changing feed delivery time from 0900 to 2100 h increased the amount of feed consumed within 3 h after feeding from 27 to 37% of total daily intake but did not affect daily dry matter intake. The cows fed at 2100 h had lower blood glucose at 2 h after feeding but greater blood lactate and β-hydroxybutyrate acid at 2 and 4 h after feeding than cows fed at 0900 h. These effects of feed delivery time on the 24-h patterns in blood metabolites may be caused by the greater feed intake during the 3 h after feed delivery of the cows fed at 2100 h. Daily averages of glucose, urea, lactate, and β-hydroxybutyrate acid and nonesterified fatty acids in peripheral blood were not affected by time of feeding. The change in feed delivery time did not affect milk yield and milk protein but increased milk fat percentage from 2.5 to 2.9% and milk fat yield from 0.98 to 1.20 kg/d in multiparous cows, without affecting milk fat in primiparous cows. The interactions between diet and time of feeding on daily feed intake, milk production, and blood metabolites were not significant. The effects of the time of feed delivery on the 24-h patterns in blood metabolites suggest that this time may affect peripheral nutrient availability. Results of this study suggest beneficial effects of feeding at 2100 h instead of at 0900 h on milk fat production of lactating cows, but parity appears to mediate this effect.     

Carnitine palmitoyltransferase I in liver of periparturient dairy cows: effects of prepartum intake, postpartum induction of ketosis, and periparturient disorders

Thirty-five multiparous Holstein cows were used to determine the role of mitochondrial carnitine palmitoyltransferase I (CPT I) in liver on peripartal adaptations of fatty acid metabolism. From dry-off to parturition, cows were fed a diet at either ad libitum (n = 17) or restricted intake (RI, 80% of calculated requirements for net energy; n = 18). After parturition, all cows were fed a lactation diet. At 4 d in milk (DIM), cows underwent a physical examination and were classified as healthy (n = 15) or having at least one periparturient disorder (PD; n = 17). Cows in the healthy group were assigned to either a control (n = 6) group or a ketosis induction (KI; n = 9) group. Cows with periparturient disorders were assigned to a third (PDC; n = 17) group. Cows in control and PDC groups were fed for ad libitum intake. Cows in KI were fed at 50% of their respective intake at d 4 postpartum starting from 5 DIM and continuing to signs of clinical ketosis or until 14 DIM; cows then were returned to ad libitum intake. Liver was biopsied at −30 d, 1 d, at signs of clinical ketosis or 14 d, and 28 d relative to parturition. Mitochondria were isolated by differential centrifugation. Activity of CPT I was 5.4 and 7.6 nmol of palmitoylcarnitine formed per min/mg of protein for ad libitum and RI, respectively, at −30 DIM. Sensitivity of CPT I to its inhibitor, malonyl CoA, did not differ between ad libitum and RI cows. Differences in CPT I activity between ad libitum and RI were no longer significant at 1 DIM. Postpartum CPT I activity and malonyl CoA sensitivity at 1 DIM, onset of clinical ketosis or 14 DIM, and 28 DIM were not affected by prepartum intake (ad libitum vs. RI), postpartum health status (healthy vs. PD), or ketosis induction status (control vs. KI vs. PDC). Activity of CPT I was positively correlated with liver total lipid, liver triglyceride, liver triglyceride to glycogen ratio, and serum nonesterified fatty acids. Activity of CPT I and dry matter intake were not correlated. Prepartum intake affected prepartum CPT I activity but not malonyl CoA sensitivity. Neither induction of primary ketosis nor periparturient disorders greatly affected CPT I activity or sensitivity, which indicates that alterations of CPT I may not be a major factor in the etiology of primary ketosis or other periparturient disorders.     

Efficacy of a novel whey protein gel complex to increase the unsaturated fatty acid composition of bovine milk fat

A novel whey protein emulsion gel (WPEG) complex was developed to protect dietary unsaturated fatty acids from rumen biohydrogenation with the goal of modifying the fatty acid composition of milk fat. Three experiments were conducted with WPEG complexes made from either whey protein concentrate containing 80% crude protein, whey protein isolate, or whey protein concentrate high-gel capacity. Each experiment lasted 3 wk. All cows received a basal total mixed ration (TMR). During wk 1 and 3, all cows received only the TMR. During wk 2, 3 control cows received 330 g/d of soybean oil added to the TMR, and the other 3 cows received 330 g/d of soybean oil in one of the WPEG complexes. During wk 2, C18:2 increased from 3.29 to 5.88 g/100 g of fat in Experiment 1, 2.91 to 7.42 g/100 g of fat in Experiment 2, and 3.57 to 6.56 g/100 g of fat in Experiment 3 for WPEG cows. Fatty acid C18:3 increased from 0.51 to 0.84, 0.52 to 1.15, and 0.51 to 0.97 g/100 g of fat for Experiments 1, 2, and 3, respectively, for WPEG cows. Higher proportions of C18:1 trans-9 in milk fat of control cows compared with WPEG cows were seen in all experiments. The proportion of C18:1 trans-11 was also higher in control cows in Experiments 1 and 2, but not in Experiment 3. The WPEG complexes successfully protected unsaturated fatty acids from rumen biohydrogenation and resulted in an increase in the unsaturated fatty acid composition of milk fat produced by Holstein cows without increasing the trans 18-carbon monoenes.     

Effect of unsaturated fatty acids and triglycerides from soybeans on milk fat synthesis and biohydrogenation intermediates in dairy cattle

Increased rumen unsaturated fatty acid (FA) load is a risk factor for milk fat depression. This study evaluated if increasing the amount of unsaturated FA in the diet as triglycerides or free FA affected feed intake, yield of milk and milk components, and feed efficiency. Eighteen Holstein cows (132 ± 75 d in milk) were used in a replicated 3 × 3 Latin square design. Treatments were a control (CON) diet, or 1 of 2 unsaturated FA (UFA) treatments supplemented with either soybean oil (FA present as triglycerides; TAG treatment) or soybean FA distillate (FA present as free FA; FFA treatment). The soybean oil contained a higher concentration of cis-9 C18:1 (26.0 vs. 11.8 g/100 g of FA) and lower concentrations of C16:0 (9.6 vs. 15.0 g/100 g of FA) and cis-9,cis-12 C18:2 (50.5 vs. 59.1 g/100 g of FA) than the soybean FA distillate. The soybean oil and soybean FA distillate were included in the diet at 2% dry matter (DM) to replace soyhulls in the CON diet. Treatment periods were 21 d, with the final 4 d used for sample and data collection. The corn silage- and alfalfa silage-based diets contained 23% forage neutral detergent fiber and 17% crude protein. Total dietary FA were 2.6, 4.2, and 4.3% of diet DM for CON, FFA, and TAG treatments, respectively. Total FA intake was increased 57% for UFA treatments and was similar between FFA and TAG. The intakes of individual FA were similar, with the exception of a 24 g/d lower intake of C16:0 and a 64 g/d greater intake of cis-9 C18:1 for the TAG compared with the FFA treatment. Compared with CON, the UFA treatments decreased DM intake (1.0 kg/d) but increased milk yield (2.2 kg/d) and milk lactose concentration and yield. The UFA treatments reduced milk fat concentration, averaging 3.30, 3.18, and 3.11% for CON, FFA, and TAG treatments, respectively. Yield of milk fat, milk protein, and 3.5% fat-corrected milk remained unchanged when comparing CON with the UFA treatments. No differences existed in the yield of milk or milk components between the FFA and TAG treatments. The UFA treatments increased feed efficiency (energy-corrected milk/DM intake), averaging 1.42, 1.53, and 1.48 for CON, FFA, and TAG treatments, respectively. Although milk fat yield was not affected, the UFA treatments decreased the yield of de novo (<16-carbon) synthesized FA (40 g/d) and increased the yield of preformed (>16-carbon) FA (134 g/d). Yield of FA from both sources (16-carbon FA) was reduced by the UFA treatments but to a different extent for FFA versus TAG (72 vs. 100 g/d). An increase was detected in the concentration of trans-10 C18:1 and a trend for an increase in trans-10,cis-12 C18:2 and trans-9,cis-11 C18:2 for the UFA treatments compared with CON. Under the dietary conditions tested, UFA treatments supplemented at 2% diet DM as either soybean FA distillate or soybean oil increased milk yield but did not effectively cause a reduction in milk fat yield, with preformed FA replacing de novo synthesized FA in milk fat. Further research is required to determine if the response to changes in dietary free and esterified FA concentrations is different in diets that differ in their risk for milk fat depression.     

Effects of rumen-protected choline and monensin on milk production and metabolism of periparturient dairy cows

Choline and monensin may be supplemented during the transition period with the objectives of aiding in fat metabolism and improving energy balance, respectively. The objectives of this study were to determine the effects of supplementing rumen-protected choline (RPC) and monensin in a controlled-release capsule (CRC) on metabolism, dry matter intake, milk production, and liver function in transition dairy cattle. Three weeks before expected calving, 182 Holsteins were randomly assigned to receive one of the following: a monensin CRC, 56 g/d of RPC until 28 d in milk, CRC + RPC, or neither supplement (control). Blood samples were collected at enrollment, 1 wk before calving, and in the first and second weeks after calving. Liver biopsies were obtained from multiparous cows randomly selected from each treatment group within 24 h and again 3 wk postpartum. Daily milk production was recorded through 60 d in milk. There were no interactions of the effects of RPC and CRC on any of the outcomes measured. Overall, cows that received RPC produced 1.2 kg/d more milk in the first 60 d of lactation, but this effect was attributable to an increase in milk production of 4.4 kg/d among cows with a body condition score ≥4 at 3 wk before calving; fat cows that received RPC ate 1.1 kg of DM/d more from wk 3 before calving through wk 4 after calving. Monensin supplementation significantly increased serum concentrations of glucose and urea, lowered concentrations of β-hydroxybutyric acid and aspartate aminotransferase in the peripartum period, and increased liver glycogen content at 3 wk into lactation. The metabolic effects of CRC are consistent with previous studies, and the effects on liver are novel. The mechanism by which RPC increased milk production was not revealed in this study and merits further research.     

Effects of rumen-protected choline and dry propylene glycol on feed intake and blood parameters for Holstein dairy cows in early lactation

A 6 × 6 Latin square design was used to test 3 sets of comparisons simultaneously to study response in dry matter intake, milk yield, and blood parameters to propylene glycol (PG) supplementation delivered by 2 methods [incorporating PG into the total mixed ration (TMR) vs. top dressing; comparison I]; individual or combined dietary choline and PG supplementation as a 2 × 2 factorial (comparison II); or increasing amounts of dietary choline (comparison III). Six multiparous (lactation number = 1.5 ± 0.8 SD) Holstein dairy cows were at 41 d in milk (± 9 SD) at the start of the experiment. Propylene glycol used was a dry product containing 65% PG, and choline was a rumen-protected choline product (RPC; estimated to be 50% rumen-protected) containing 50% choline chloride. In comparison I, treatments compared were 1) control: no PG; 2) PG-TMR: 250 g/d of dry PG (corresponding to 162.5 g/d of PG) incorporated into the TMR; and 3) PG-top dress: 250 g/d of dry PG top-dressed onto the TMR. In comparison II, treatments compared were 1) control: no PG and no RPC; 2) PG: 250 g/d of dry PG incorporated into the TMR; 3) RPC: 50 g/d of RPC top-dressed onto the TMR; and 4) PG + RPC: combination of treatments 2 and 3. In comparison III, treatments compared were 0, 25, and 50 g/d of RPC top-dressed onto the TMR. Each experimental period lasted 10 d with 9 d of adaptation followed by 1 d of serial blood sampling. Dry matter intake and milk yield were recorded daily. During the serial blood sampling, jugular blood was sampled every 20 min for the first 4 h and at 8 and 12 h after treatment administration. Results obtained from comparison I showed that feeding 250 g/d of PG as a dry product decreased plasma β-hydroxybutyrate (BHBA) concentration (mean ± SEM) from 701 ± 81 (control) to 564 ± 76 μmol/L without affecting serum insulin, plasma glucose, or plasma nonesterified fatty acid concentrations. Top-dressing PG decreased plasma BHBA concentrations more than by incorporating it into the TMR [527 vs. 601 μmol/L (± 81 pooled SEM)]. Results obtained from comparison II showed that supplementing choline as RPC, PG, or both had no effect on dry matter intake, milk yield, or any of the blood parameters measured. Results obtained from comparison III showed that milk yield tended to increase linearly with increasing amounts of dietary choline as RPC. We concluded that feeding PG as a dry product reduced plasma BHBA concentration but top-dressing PG was more efficient at reducing plasma BHBA level than incorporating PG into the TMR. Dietary choline as RPC tended to increase milk yield linearly. However, a combined effect of dietary PG and choline was not evident and therefore not beneficial.     

Effects of fat and methionine hydroxy analog on prevention or alleviation of fatty liver induced by feed restriction

Two trials were conducted to examine the effects of supplemental methionine, provided as methionine hydroxy analog 13 g/d), or fat (454 g of calcium salts of long-chain fatty acids/d) on hepatic triglyceride concentration. In the first experiment, methionine hydroxy analog or fat was fed during feed restriction to determine if hepatic triglyceride accumulation is affected. The objective of the second experiment was to determine if feeding fat or methionine hydroxy analog influences the rate of triglyceride depletion from the liver of cows in positive energy balance following the induction of fatty liver by feed restriction. In experiment 1, feeding methionine hydroxy analog decreased plasma glucose, increased plasma nonesterified fatty acids, and had no effect on liver triglyceride. Feeding fat increased plasma nonesterified fatty acids and increased hepatic triglyceride during the 10-d feed restriction period. In experiment 2, feeding fat decreased the rate of triglyceride depletion from liver when cows were allowed to resume ad libitum consumption of feed; methionine hydroxy analog had no effect. Results of these studies indicate that feeding supplemental fat or methionine hydroxy analog at levels tested does not prevent or alleviate fatty liver induced by feed restriction.     

Diet supplementation with fish oil and sunflower oil to increase conjugated linoleic acid levels in milk fat of partially grazing dairy cows

The objective of this study was to determine the long-term effect on milk conjugated linoleic acid (cis-9, trans-11 CLA) of adding fish oil (FO) and sunflower oil (SFO) to the diets of partially grazing dairy cows. Fourteen Holstein cows were divided into 2 groups (7 cows/treatment) and fed either a control or oil-supplemented diet for 8 wk while partially grazing pasture. Cows in group 1 were fed a grain mix diet (8.0 kg/d, DM basis) containing 400 g of saturated animal fat (control). Cows in the second group were fed the same grain mix diet except the saturated animal fat was replaced with 100 g of FO and 300 g of SFO. Cows were milked twice a day and milk samples were collected weekly throughout the trial. Both groups grazed together on alfalfa-based pasture ad libitum and were fed their treatment diets after the morning and afternoon milking. Milk production (30.0 and 31.2 kg/d), milk fat percentages (3.64 and 3.50), milk fat yield (1.08 and 1.09 kg/d), milk protein percentages (2.97 and 2.88), and milk protein yield (0.99 and 0.91 kg/d) for diets 1 and 2, respectively, were not affected by the treatment diets. The concentrations of cis-9, trans-11 CLA (1.64 vs. 0.84 g/100 g of fatty acids) and vaccenic acid (5.11 vs. 2.20 g/100 g of fatty acids) in milk fat were higher for cows fed the oil-supplemented diet over the 8 wk of oil supplementation. The concentration of cis-9, trans-11 CLA in milk fat reached a maximum (1.0 and 1.64 g/100 g of fatty acids for diets 1 and 2, respectively) in wk 1 for both diets and remained relatively constant thereafter. The concentration of vaccenic acid in milk fat followed the same temporal pattern as cis-9, trans-11 CLA. In conclusion, supplementing the diet of partially grazing cows with FO and SFO increased the milk cis-9, trans-11 CLA content, and that increase remained relatively constant after 1 wk of oil supplementation.     

Noninvasive detection of hepatic lipidosis in dairy cows with calibrated ultrasonographic image analysis

The aim was to test the accuracy of calibrated digital analysis of ultrasonographic hepatic images for diagnosing fatty liver in dairy cows. Digital analysis was performed by means of a novel method, computer-aided ultrasound diagnosis (CAUS), previously published by the authors. This method implies a set of pre- and postprocessing steps to normalize and correct the transcutaneous ultrasonographic images. Transcutaneous hepatic ultrasonography was performed before surgical correction on 151 German Holstein dairy cows (mean ± standard error of the means; body weight: 571 ± 7 kg; age: 4.9 ± 0.2 yr; DIM: 35 ± 5) with left-sided abomasal displacement. Concentration of triacylglycerol (TAG) was biochemically determined in liver samples collected via biopsy and values were considered the gold standard to which ultrasound estimates were compared. According to histopathologic examination of biopsies, none of the cows suffered from hepatic disorders other than hepatic lipidosis. Hepatic TAG concentrations ranged from 4.6 to 292.4 mg/g of liver fresh weight (FW). High correlations were found between the hepatic TAG and mean echo level (r = 0.59) and residual attenuation (ResAtt; r = 0.80) obtained in ultrasonographic imaging. High correlation existed between ResAtt and mean echo level (r = 0.76). The 151 studied cows were split randomly into a training set of 76 cows and a test set of 75 cows. Based on the data from the training set, ResAtt was statistically selected by means of stepwise multiple regression analysis for hepatic TAG prediction (R² = 0.69). Then, using the predicted TAG data of the test set, receiver operating characteristic analysis was performed to summarize the accuracy and predictive potential of the differentiation between various measured hepatic TAG values, based on TAG predicted from the regression formula. The area under the curve values of the receiver operating characteristic based on the regression equation were 0.94 (<50 vs. ≥50 mg of TAG/g of FW), 0.83 (<100 vs. ≥100 mg of TAG/g of FW), and 0.97 (<50 vs. ≥100 mg of TAG/g of FW). The CAUS methodology and software for digitally analyzing liver ultrasonographic images is considered feasible for noninvasive screening of fatty liver in dairy herd health programs. Using the single parameter linear regression equation might be ideal for practical applications.     

Effects of prilled fatty acids and calcium salts of fatty acids on rumen fermentation, nutrient digestibilities, milk production, and milk composition

Lactating Holstein cows averaging 193 d postpartum and fitted with rumen cannulae were used in two experiments to investigate the effects of supplementing Ca salts of fatty acids or prilled fatty acids to the diet on fermentation in the rumen, apparent total tract nutrient digestibility, milk production, and milk composition. Cows were fed ad libitum total mixed diets consisting of 45% concentrate and 55% forage. Treatments in Experiment 1 were: 1) control, 2) control plus 680 g/cow per d of Ca salts of fatty acids, 3) control plus 680 g/cow per d of prilled fatty acids, or 4) control plus 907 g/cow per d of prilled fatty acids. Treatments in Experiment 2 were: 1) control, 2) control plus 553 g/cow per d of Ca salts of fatty acids, or 3) control plus 454 g/cow per d of prilled fatty acids. Data suggest that Ca salts of fatty acids and prilled fatty acids are inert in the rumen and do not greatly alter fermentation in the rumen, apparent total tract digestibilities of DM, organic matter, ADF, NDF, and CP, or milk composition when fed at recommended amounts of 3 to 4% of the DM intake. The fact that milk production was not increased in these experiments by feeding Ca salts of fatty acids and prilled fatty acids may be attributed to the use of medium to low producing cows that were past the peak of milk production. Additional experiments are needed to obtain information about feeding these sources of supplemental fat to high producing cows during the early stages of lactation.