Effects of short- and long-chain fatty acids on the expression of stearoyl-CoA desaturase and other lipogenic genes in bovine mammary epithelial cells.

Stearoyl-CoA desaturase (SCD) in the bovine mammary gland introduces a cis-double bond at the Δ9 position in a wide range of fatty acids (FA). Several long-chain polyunsaturated fatty acids (PUFA) inhibit expression of SCD, but information on the effect of short-chain fatty acids on mammary SCD expression is scarce. We used a bovine mammary cell line (MAC-T) to assess the effect of acetic acid (Ac) and ß-hydroxybutyric acid (BHBA) in comparison with the effect of various long-chain fatty acids on the mRNA expression of the lipogenic enzymes SCD, acetyl-CoA carboxylase (ACACA), fatty acid synthase (FASN) and their associated gene regulatory proteins sterol regulatory element binding transcription factor 1 (SREBF1), insulin-induced gene 1 protein (INSIG1) and peroxisome proliferator-activated receptor alpha (PPARA)and peroxisome proliferator-activated receptor delta (PPARD) by quantitative real-time PCR. MAC-T cells were treated for 12 h without FA additions (CON) or with either 5 mM Ac, 5 mM BHBA, a combination of 5 mM Ac + 5 mM BHBA, 100 µM C16:0, 100 µM C18:0, 100 µM C18:1 cis-9, 100 µM C18:1 trans-11, 100 µM C18:2 cis-9,12 or 100 µM C18:3 cis-9,12,15. Compared with control, mRNA expression of SCD1 was increased by Ac (+61%) and reduced by C18:1 cis-9 (-61%), C18:2 cis-9,12 (-84%) and C18:3 cis-9,12,15 (-88%). In contrast to native bovine mammary gland tissue, MAC-T cells did not express SCD5. Expression of ACACA was increased by Ac (+44%) and reduced by C18:2 cis-9,12 (-48%) and C18:3 cis-9,12,15 (-49%). Compared with control, FASN expression was not significantly affected by the treatments. The mRNA level of SREBF1 was not affected by Ac or BHBA, but was reduced by C18:1 cis-9 (-44%), C18:1 trans-11 (-42%), C18:2 cis-9,12 (-62%) and C18:3 cis-9,12,15 (-68%) compared with control. Expression of INSIG1 was downregulated by C18:0 (-37%), C18:1 cis-9 (-63%), C18:1 trans-11 (-53%), C18:2 cis-9,12 (-81%) and C18:3 cis-9,12,15 (-91%). Both PPARA and PPARD expression were not significantly affected by the treatments. Our results show that Ac upregulated mRNA expression of SCD1 and ACACA in MAC-T cells. The opposite effect of the PUFA C18:2 cis-9,12 and C18:3 cis-9,12,15 on the these genes and the failure of Ac to mimic the PUFA-inhibited SREBF1 and INSIG1 mRNA expression, suggest that Ac can stimulate mammary lipogenesis via a transcriptional regulatory mechanism different from PUFA.

Response profiles of enteric methane emissions and lactational performance during habituation to dietary coconut oil in dairy cows.

Dietary coconut oil (CNO) can reduce dry matter intake (DMI), enteric methane (eCH(4)) emissions, and milk fat yield of lactating cows. The goals of this research were to examine responses to different CNO concentrations during the habituation period (34-d) and to evaluate temporal patterns of DMI, eCH(4), and milk fat yield. Treatment diets contained (dry basis): 0.0% (CNO0), 1.3% (CNO1.3), 2.7% (CNO2.7), 3.3% (CNO3.3), or 4.0% CNO (CNO4). In experiment 1, 12 primi- or small secundiparous cows were housed in individual, environmentally controlled rooms and fed CNO0, CNO1.3, CNO2.7, or CNO4. Measurements included DMI, eCH(4), and milk yield and composition. Due to a precipitous drop in DMI (26%), cows fed CNO4 were replaced with cows fed CNO3.3 following d 10. Dietary CNO of 2.7% or more reduced eCH(4) emissions. Reduction was greater with increased CNO and during the first than the second half of the day. Simultaneously, decline in DMI of cows fed CNO2.7, CNO3.3, or CNO4 was increasingly precipitous with increased CNO concentration. Total-tract neutral detergent fiber (NDF) digestibility during wk 5 was reduced in cows fed CNO2.7 or CNO3.3, which in part explained concomitantly reduced eCH(4)/DMI. In addition, milk fat yield was depressed at an increasing rate in cows fed CNO2.7, CNO3.3, and CNO4. In experiment 2, DMI was measured individually in 12 multiparous cows during habituation to CNO0, CNO1.3, CNO2.7, or CNO3.3 for 21 d before relocation to individual, environmentally controlled rooms. Dietary CNO2.7 or CNO3.3 reduced DMI by d 4 and total-tract NDF digestibility during wk 5. Relocation to individual rooms was associated with a 15% reduction in DMI, which was not affected by treatment. Results showed that 2.7% or more dietary CNO reduced eCH(4) and DMI, caused milk fat depression, and decreased NDF digestibility.

Enteric methane emissions and lactational performance of Holstein cows fed different concentrations of coconut oil.

To determine if dietary medium-chain fatty acids (FA; C(8) to C(14)) may mitigate enteric methane emissions, 24 cows were blocked by body size (n=2) and randomly assigned to 1 sequence of dietary treatments. Diets were fed for 35 d each in 2 consecutive periods. Diets differed in concentrations of coconut oil (CNO; ~75% medium-chain FA): 0.0 (control) or 1.3, 2.7, or 3.3% CNO, dry matter basis. The control diet contained 50% forage (74% from corn silage), 16.5% crude protein (60% from rumen-degradable protein), 34% neutral detergent fiber (NDF; 71% from forage), and 28% starch, dry matter basis. Data and sample collections were from d 29 to 35 in environmentally controlled rooms to measure methane (CH(4)) production. Methane emitted was computed from the difference in concentrations of inlet and outlet air and flux as measured 8 times per day. Control cows emitted 464 g of CH(4)/d, consumed 22.9 kg of DM/d, and produced 34.8 kg of solids-corrected milk/d and 1.3 kg of milk fat/d. Treatment with 1.3, 2.7, or 3.3% dietary CNO reduced CH(4) (449, 291, and 253 g/d, respectively), but concomitantly depressed dry matter intake (21.4, 17.9, and 16.2 kg/d, respectively), solids-corrected milk yield (36.3, 28.4, and 26.8 kg/d, respectively), and milk fat yield (1.4, 0.9, and 0.9 kg/d, respectively). The amount of NDF digested in the total tract decreased with increased dietary CNO concentrations; thus, CH(4) emitted per unit of NDF digested rose from 118 to 128, 153, and 166 g/kg across CNO treatments. Dietary CNO did not significantly affect apparent digestibility of CP but increased apparent starch digestibility from 92 to 95%. No FA C(10) or shorter were detected in feces, and apparent digestibility decreased with increasing FA chain length. Coconut oil concentrations of 2.7 or 3.3% decreased yields of milk FA C(14). The highest milk fat concentration (3.69%; 1.3% CNO) was due to the greatest yields of C(12) to C(16) milk FA. Milk FA concentrations of C(18:2 trans-10,cis-12) were related to increased dietary CNO concentrations and presumably to depressed ruminal NDF digestion. Moderate dietary CNO concentrations (e.g., 1.3%) may benefit lactational performance; however, CNO concentrations greater than or equal to 2.7% depressed dry matter intake, milk yield, milk fat yield, and NDF utilization. If mitigation of enteric CH(4) emissions is due to decreased digestion of dietary NDF, then this will lessen a major advantage of ruminants compared with nonruminants in food-production systems. Thus, CNO has limited use for enteric CH(4) mitigation in lactating dairy cows.

A simple analytical and experimental procedure for selection of reference genes for reverse-transcription quantitative PCR normalization data.

Variation in cellular activity in a tissue induces changes in RNA concentration, which affects the validity of gene mRNA abundance analyzed by reverse transcription quantitative PCR (RT-qPCR). A common way of accounting for such variation consists of the use of reference genes for normalization. Programs such as geNorm may be used to select suitable reference genes, although a large set of genes that are not co-regulated must be analyzed to obtain accurate results. The objective of this study was to propose an alternative experimental and analytical protocol to assess the invariance of reference genes in porcine mammary tissue using mammary RNA and DNA concentrations as correction factors. Mammary glands were biopsied from 4 sows on d 110 of gestation (prepartum), on d 5 (early) and 17 (peak) of lactation, and on d 5 after weaning (postweaning). Relative expression of 7 potential reference genes, API5, MRPL39, VAPB, ACTB, GAPDH, RPS23, and MTG1, and one candidate gene, SLC7A1, was quantified by RT-qPCR using a relative standard curve approach. Variation in gene expression levels, measured as cycles to threshold at each stage of mammary physiological activity, was tested using a linear mixed model fitting RNA and DNA concentrations as covariates. Results were compared with those obtained with geNorm analysis, and genes selected by each method were used to normalize SLC7A1. Quantified relative mRNA abundance of GAPDH and MRPL39 remained unchanged across stages of mammary physiological activity after accounting for changes in tissue RNA and DNA concentration. In contrast, geNorm analysis selected MTG1, MRPL39, and VAPB as the best reference genes. However, when target gene SLC7A1 was normalized with genes selected either based on our proposed protocol or by geNorm, fold changes in mRNA abundance did not differ. In conclusion, the proposed analytical protocol assesses expression invariance of potential reference genes by accounting for variation in tissue RNA and DNA concentrations and thus represents an alternative method to select suitable reference genes for RT-qPCR analysis.

Effect of intensified feeding of heifer calves on growth, pubertal age, calving age, milk yield, and economics.

The objective of this study was to determine if increasing the energy and protein intake of heifer calves would affect growth rates, age at puberty, age at calving, and first lactation milk yield. A second objective was to perform an economic analysis of this feeding program using feed costs, number of nonproductive days, and milk yield data. Holstein heifer calves born at the Michigan State Dairy Cattle Teaching and Research Center were randomly assigned to 1 of 2 dietary treatments (n=40/treatment) that continued from 2 d of age until weaning at 42 d of age. The conventional diet consisted of a standard milk replacer [21.5% crude protein (CP), 21.5% fat] fed at 1.2% of body weight (BW) on a dry matter basis and starter grain (19.9% CP) to attain 0.45 kg of daily gain. The intensive diet consisted of a high-protein milk replacer (30.6% CP, 16.1% fat) fed at 2.1% of BW on a dry matter basis and starter grain (24.3% CP) to achieve 0.68 kg of daily gain. Calves were gradually weaned from milk replacer by decreasing the amount offered for 5 and 12 d before weaning for the conventional and intensive diets, respectively. All calves were completely weaned at 42 d of age and kept in hutches to monitor individual starter consumption in the early postweaning period. Starting from 8 wk of age, heifers on both treatments were fed and managed similarly for the duration of the study. Body weight and skeletal measurements were taken weekly until 8 wk of age, and once every 4 wk thereafter until calving. Calves consuming the intensive diet were heavier, taller, and wider at weaning. The difference in withers height and hip width was carried over into the early post-weaning period, but a BW difference was no longer evident by 12 wk of age. Calves fed the intensive diet were younger and lighter at the onset of puberty. Heifers fed the high-energy and protein diet were 15 d younger at conception and 14 d younger at calving than heifers fed the conventional diet. Body weight after calving, daily gain during gestation, withers height at calving, body condition score at calving, calving difficulty score, and calf BW were not different. Energy-corrected, age-uncorrected 305-d milk yield was not different, averaging 9,778 kg and 10,069 kg for heifers fed the conventional and intensive diets, respectively. However, removing genetic variation in milk using parent average values as a covariate resulted in a tendency for greater milk from heifers fed the intensive diet. Preweaning costs were higher for heifers fed the intensive diet. However, total costs measured through first lactation were not different. Intensified feeding of calves can be used to decrease age at first calving without negatively affecting milk yield or economics.

Intramammary infusion of leptin decreases proliferation of mammary epithelial cells in prepubertal heifers.

High energy intake and excessive body fatness impair mammogenesis in prepubertal ruminants. High energy intake and excessive fatness also increase serum leptin. Our objective was to determine if an infusion of leptin decreases proliferation of mammary epithelial cells of prepubertal heifers in vivo. Ovine leptin at 100 microg/ quarter per d with or without 10 microg of insulin-like growth factor (IGF)-I was infused via the teat canal into mammary glands of prepubertal dairy heifers; contralateral quarters were used as controls. After 7 d of treatment, bromodeoxyuridine was infused intravenously and heifers were slaughtered approximately 2 h later. Tissue from 3 regions of the mammary parenchyma was collected and immunostained for bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (Ki-67), and caspase-3. Leptin decreased the number of mammary epithelial cells in the S-phase of the cell cycle by 48% in IGF-I-treated quarters and by 19% in saline-treated quarters. Leptin did not alter the number of mammary epithelial cells within the cell cycle, as indicated by Ki-67 labeling. Caspase-3 immunostaining within the mammary parenchyma was very low in these heifers, but leptin significantly increased labeling in saline-treated quarters. Leptin enhanced SOCS-3 expression in IGF-I-treated quarters but did not alter SOCS-1 or SOCS-5 expression. We conclude that a high concentration of leptin in the bovine mammary gland reduces proliferation of mammary epithelial cells. The reduced proliferation is accompanied by an increase in SOCS-3 expression, suggesting a possible mechanism for leptin inhibition of IGF-I action. Whether leptin might be a physiological regulator of mammogenesis remains to be determined.

Effects of feeding prepubertal heifers a high-energy diet for three, six, or twelve weeks on feed intake, body growth, and fat deposition.

The objective was to determine the effects of feeding prepubertal dairy heifers a high-energy diet for a duration of 0, 3, 6, or 12 wk on feed intake, growth, and fat deposition. We also used feed composition, daily intake, and body growth data to evaluate the nutritional model of the 2001 National Research Council (NRC) Nutrient Requirements of Dairy Cattle. Holstein heifers (age = 11 wk; body weight = 107 +/- 1 kg) were assigned to 1 of 4 treatments (n = 16/treatment) designated H0, H3, H6, and H12 and fed a low-energy diet for 12, 9, 6, or 0 wk, followed by a high-energy diet for 0, 3, 6, or 12 wk, respectively. Four heifers were killed initially (11 wk of age) and 64 heifers were killed at the end of the treatment period (23 wk of age). The low-energy diet was formulated to achieve 0.6 kg of average daily gain and contained 16% crude protein, and 45% neutral detergent fiber. The high-energy diet was formulated to achieve an average daily gain of 1.2 kg and contained 18% crude protein and 23% neutral detergent fiber. Actual daily gains averaged over the 12-wk treatment period were 0.64, 0.65, 0.83, and 1.09 kg for the H0, H3, H6, and H12 groups, respectively. Body weight, withers height, hip width, carcass weight, liver weight, and perirenal fat increased in heifers fed a high-energy diet for a longer duration. In addition, percentage of fat increased and percentage of protein decreased in rib sections with a longer duration on the high-energy diet. Uterine and ovarian weights adjusted for body weight decreased when heifers were fed the high-energy diet for a longer duration. The 2001 NRC underestimated dry matter intake of the high-energy diet and overestimated dry matter intake of the low-energy diet. On the basis of actual intakes of each diet, the NRC slightly underestimated gain for the low-energy diet and overestimated gain by 40% for the high-energy diet. The likely explanation for this is that the NRC underestimated the proportion of gain that was fat in the heifers fed the high-energy diet and therefore predicted more body gain per unit of energy intake. We concluded that feeding a high-energy diet for a short duration altered body growth and fat deposition in a time-dependent, linear manner consistent with feeding a high-energy diet for a long duration.

Effects of feeding prepubertal heifers a high-energy diet for three, six, or twelve weeks on mammary growth and composition.

The experimental objective was to determine the effects of feeding prepubertal dairy heifers a high-energy diet for 3, 6, or 12 wk on mammary growth and composition. Holstein heifers (age = 11 wk; body weight = 107 +/- 1 kg) were assigned to 1 of 4 treatments (n = 16/ treatment). The treatment period lasted 12 wk and treatments were H0 (low-energy diet fed for 12 wk, with no weeks on the high-energy diet); H3 (low-energy diet fed for 9 wk, followed by the high-energy diet for 3 wk); H6 (low-energy diet fed for 6 wk, followed by the high-energy diet for 6 wk); and H12 (high-energy diet for all 12 wk). The low- and high-energy diets were formulated to achieve 0.6 and 1.2 kg of average daily gain, respectively. Heifers were slaughtered at 23 wk of age and mammary tissue was collected. A longer duration of feeding the high-energy diet increased total mass of the mammary gland, extraparenchymal fat, and intraparenchymal fat, but did not alter the mass of fat-free parenchymal tissue. When adjusted for carcass weight to reflect differences in physical maturity, the mass of fat-free parenchymal tissue decreased in a linear fashion with a longer duration on the high-energy diet. Total masses of mammary parenchymal DNA and RNA were not different. However, after adjustment for carcass weight, the masses of DNA and RNA decreased as heifers were fed the high-energy diet for a longer duration. The percentages of epithelium, stroma, and lumen, the number of epithelial structures, and the developmental scores of mammary parenchymal tissue were not different among treatments. However, the percentage of proliferating epithelial cells in the terminal ductal units, as indicated by Ki-67 labeling, decreased as heifers were fed the high-energy diet for a longer duration. We concluded that feeding prepubertal heifers a high-energy diet for a longer duration resulted in a linear decrease in both the percentage of mammary epithelial cells that were proliferating and in the mass of fat-free mammary parenchyma per unit of carcass. High-energy feeding hastens puberty and, in this study, decreased mammary epithelial cell proliferation in areas of active ductal expansion. These data are consistent with the idea that feeding heifers a high-energy diet will reduce mammary parenchymal mass at puberty.

Short communication: intramammary infusion of IGF-I increases bromodeoxyuridine labeling in mammary epithelial cells of prepubertal heifers.

When dairy heifers are fed to gain more than 900 g of body weight/d, they have less mammary parenchymal DNA at puberty but more insulin-like growth factor-I (IGF-I) in serum. This negative relationship between serum IGF-I concentration and mammary epithelial cell proliferation is in disagreement with the extensively reported role of IGF-I as a stimulator of mammary epithelial cell proliferation. Despite the large body of evidence suggesting that an increase in IGF-I concentration should lead to an increase in mammary epithelial cell proliferation of prepubertal heifers, it had not been previously tested. Our objective was to determine if intramammary infusions of IGF-I would stimulate mammogenesis in prepubertal heifers in vivo. After 7 d of treatment, bromodeoxyuridine was infused intravenously and heifers were slaughtered 3 h later. Samples from 3 regions of the mammary parenchyma were collected, fixed, sliced, and incubated with bromodeoxyuridine monoclonal antibody to identify cells in the S-phase of the cell cycle. Intramammary infusion of IGF-I increased the percentage of epithelial cells in the S-phase by 52% (6.4 vs. 4.2%, +/- 0.3%). Proliferation was similar in all 3 parenchymal regions, and the response to IGF-I was similar in each region. We conclude that local IGF-I increases proliferation of mammary parenchymal epithelial cells in prepubertal heifers.

Effect of increasing energy and protein intake on body growth and carcass composition of heifer calves.

The objective was to determine whether increased energy and protein intake between 2 and 14 wk of age would increase growth rates of heifer calves without fattening. At 2 wk of age, Holstein heifer calves were assigned to 1 of 4 treatments in a 2 x 2 factorial arrangement with 2 levels of protein and energy intake (moderate [M]; high [H]) in period 1 (2 to 8 wk of age) by 2 levels of protein and energy intake (low [L]; high [H]) in period 2 (8 to 14 wk of age) to produce similar initial BW for all 4 treatments. Treatments were ML, MH, HL, and HH, indicating moderate or high energy and protein intake during the first period and low or high intake during the second period. The M diet consisted of a standard milk replacer (21.3% CP, 21.3% fat) fed at 1.1% of BW on a DM basis and a 16.5% CP grain mix fed at restricted intake to promote 400 g of average daily gain (ADG), whereas the L diet consisted only of the grain mix. The H diet consisted of a high-protein milk replacer (30.3% CP, 15.9% fat) fed at 2% of BW on a DM basis and a 21.3% CP grain mix available ad libitum. Calves were weaned gradually from milk replacer by 7 wk and slaughtered at 8 (n = 11) or 14 wk of age (n = 41). In periods 1 and 2, ADG and the gain:feed ratio were greater for calves fed the H diet. Calves fed the H diet were taller after both periods 1 and 2. No difference was observed in carcass composition at 8 wk, but at 14 wk calves fed MH and HH had less water and more fat than calves fed ML and HL. Plasma IGF-I concentrations were greatest for calves fed the H diet during either period. Plasma leptin concentrations were increased in calves fed the H diet during period 1 from 4 to 6 wk of age. Increasing energy and protein intake from 2 to 8 wk and 8 to 14 wk of age increased BW, withers height, and gain:feed ratio. Calves fed the H diet from 8 to 14 wk of age had more body fat than calves fed the L diet. Increased energy and protein intake can increase the rate of body growth of heifer calves and potentially reduce rearing costs.

Effect of increasing energy and protein intake on mammary development in heifer calves.

The objective of this study was to determine if increased energy and protein intake from 2 to 14 wk of age would affect mammary development in heifer calves. At 2 wk of age, Holstein heifer calves were assigned to 1 of 4 treatments in a 2 x 2 factorial arrangement with 2 levels of protein and energy intake (moderate, M; high, H) in period 1 (2 to 8 wk of age) and 2 levels of protein and energy intake (low, L; high, H) in period 2 (8 to 14 wk of age), so that mean initial body weights were approximately equal for all 4 treatments (ML, MH, HL, and HH). The M diet in period 1 consisted of a standard milk replacer (21.3% CP, 21.3% fat) fed at 1.1% of BW on a DM basis and a 16.5% CP grain mix fed at restricted intake to promote 400 g of daily gain, whereas the L diet in period 2 consisted only of the grain mix. The H diet in period 1 consisted of a high-protein milk replacer (30.3% CP, 15.9% fat) fed at 2.0% of body weight on a DM basis and a 21.3% CP grain mix available ad libitum. In period 2, the H diet consisted of just the 21.3% grain mix. Calves were gradually weaned from milk replacer by 7 wk and slaughtered at 8 (n = 11) or 14 wk of age (n = 41). Parenchyma from the distal region, midgland, and proximal region relative to the teat from one half of the udder was collected, fixed, and embedded in paraffin. The other half of the gland was used to determine parenchymal mass, protein, fat, DNA, RNA, and extraparenchymal mass. Total parenchymal tissue, parenchymal DNA, parenchymal RNA, and concentrations of DNA and RNA were higher for calves on the H diet during period 1, but were not affected by diet during period 2. Parenchymal fat percentage was increased by the H diet during period 2. The H diet increased extraparenchymal fat during both periods. The area of parenchyma occupied by epithelium was not affected by treatment, but at the end of period 2, the percentage of proliferating epithelial cells as indicated by Ki67, an marker of cell proliferation, expression was greater for calves on the M diet in period 1 compared with calves on the H diet in period 1. Diets did not influence parenchymal protein percentage or the ratio of RNA to DNA. Higher energy and protein intake from 2 to 8 wk of age increased parenchymal mass and parenchymal DNA and RNA in mammary glands of heifer calves without increasing deposition of parenchymal fat. Diet also influenced histological development of mammary parenchyma and subsequent proliferation of ductal epithelial cells. Implications of these effects for future milk production potential are unknown.

Clinical responses to intramammary endotoxin infusion in dairy cows subjected to feed restriction.

Nonpregnant, midlactation primiparous Holstein cows were fed ad libitum (n = 12) or at 80% of maintenance energy requirements (n = 12) to determine whether feed restriction influences clinical response to endotoxin-induced mastitis. After 2 wk of ad libitum or restricted feeding, one mammary quarter per cow was infused with 100 microg of endotoxin. Within 3 to 6 h of intramammary infusion, endotoxin increased mean rectal temperature, heart rate, and milk somatic cell count and immunoglobulin (IgG) concentration; and decreased blood leukocyte count and rumen motility in both restricted and ad libitum-fed cows. Mean serum and milk tumor necrosis factor-alpha (TNF-alpha) concentrations showed only modest increases following endotoxin infusion. Restricted fed cows had slightly different acute fever responses and significantly increased heart and respiration rates than ad libitum fed cows. However, feed restriction did not influence mean total leukocyte count, rumen motility, serum TNF-a concentrations or milk IgG and TNF-alpha concentrations. Thus, results of this study suggest that energy balance does not significantly alter clinical symptoms following acute endotoxin-induced mastitis, at least in midlactation cows. As such, negative energy balance may not underlie the increases in severe coliform mastitis commonly observed in periparturient dairy cows.

Effects of altering dietary cation-anion difference on calcium and energy metabolism in peripartum cows.

Our objective was to determine the effects of varying dietary cation-anion differences (DCAD: meq[(Na + K) - (Cl + S)]/100 g of dry matter) in prepartum diets on Ca, energy, and endocrine status prepartum and postpartum. Holstein cows (n = 21) and heifers (n = 34) were fed diets with varying amounts of CaCl2, CaSO4, and MgSO4 to achieve a DCAD of +15 (control), 0, or -15 meq/100 g of dry matter for the last 24 d before expected calving. Dietary Ca concentration was increased (by CaCO3 supplementation) with decreasing DCAD. Plasma ionized Ca concentrations prepartum and at calving in both cows and heifers increased with reduced DCAD in the diet. At calving, plasma ionized Ca concentration was 3.67, 3.85, and 4.35 for cows and 4.44, 4.57, and 4.62 mg/dl for heifers fed diets containing +15, 0, and -15 DCAD, respectively. All heifers had normal concentrations of plasma ionized Ca (>4 mg/dl) at calving. Also at calving, plasma concentrations ofparathyroid hormone and calcitriol were less in cows and heifers fed diets containing reduced DCAD, but the plasma concentration of hydroxyproline was not affected by diet. Prepartum dry matter intake, energy balance, and body weight gains were lower and concentration of liver triglyceride was higher for heifers but not cows fed the -15 DCAD diet. Also, nonesterified fatty acids the last week prepartum were positively correlated with liver triglyceride for heifers but not cows. Feeding of anionic salts plus CaCO3 to reduce DCAD to -15 and increase Ca in prepartum diets prevents hypocalcemia at calving in cows, but decreases prepartum dry matter intake and increases the concentration of liver triglyceride in heifers. That heifers maintained calcium homeostasis at calving regardless of diet but ate less when fed the -15 DCAD diet suggests that they should not be fed anionic salts before calving.

Effect of energy and protein density of prepartum diets on fat and protein metabolism of dairy cattle in the periparturient period.

To determine if increased nutrient density in prepartum diets improves nutrient balance of peripartum cows, we blocked 40 Holstein cows and 40 heifers by expected date of parturition and assigned them randomly within blocks to one of four treatment diets varying in density of net energy for lactation (NEL) and crude protein (CP). Diets were 1.30 Mcal of NEL/kg and 12.2% CP, 1.49 Mcal of NEL/kg and 14.2% CP, 1.61 Mcal of NEL/kg and 15.9% CP, and 1.48 Mcal of NEL/kg and 16.2% CP. These diets were fed ad libitum from 25 d prepartum until parturition, and all cows were fed the same diet after calving. Increased nutrient-density of prepartum diets did not decrease feed intake. Compared to animals fed the lowest density, those fed the highest density consumed more NEL (20 vs. 14 Mcal/d) and gained more body condition, backfat, and body weight. They also had less nonesterified fatty acids in plasma (176 vs. 233 microM) and more insulin-like growth factor-I in plasma (472 vs. 390 ng/ml) during the last 2 wk prepartum and less triglyceride in liver at parturition (0.9 vs. 1.5%, wet tissue basis). Quadratic effects of energy density were not observed, and the addition of protein in the medium energy diet had no effect. Prepartum diets did not alter any variables during lactation. In conclusion, increasing the energy and protein density up to 1.6 Mcal of NEL/kg and 16% CP in diets during the last month before parturition improves nutrient balance of cattle prepartum and decreases hepatic lipid content at parturition.

Dry cow diet, management, and energy balance as risk factors for displaced abomasum in high producing dairy herds.

The objective of this study was to determine prepartum risk factors for displaced abomasum. The design was a prospective study of 1170 multiparous Holstein cows from 67 high producing dairy herds in Michigan. Each farm was visited four times within a 6-wk period. At each visit, data on nutrition and management were collected. All multiparous cows within 35 d of projected calving were assigned a body condition score, and blood was sampled to determine the concentration of nonesterified fatty acids in plasma. A multivariable linear regression model was used to determine risk factors associated with the incidence of displaced abomasum during lactation on a herd basis. A multivariable logistic regression model with random effect was used to determine risk factors for displaced abomasum on an individual cow basis. Significant risk factors for displaced abomasum included a negative energy balance prepartum (as estimated from plasma nonesterified fatty acids), a high body condition score, suboptimal feed bunk management prepartum, prepartum diets containing > 1.65 Mcal of net energy for lactation/kg of dry matter, winter and summer seasons, high genetic merit, and low parity.

Comparison of growth hormone-releasing factor and somatotropin: lipid and glucose metabolism in dairy cows.

Lactating, primiparous Holstein cows were continuously infused for 63 d with either recombinant growth hormone-releasing factor or recombinant bST or were not infused (control). The releasing factor and bST treatments reduced fat synthesis from acetate and lipoprotein lipase activity in omental adipose tissue. Also, both treatments increased hormone-sensitive lipase activity per gram of adipose tissue and reduced fatty acid esterification per milligram of protein in adipose tissue. Protein concentration in adipose tissue with both treatments was greater than that of the control, although the protein concentration was less in adipose tissue from cows treated with the releasing factor than from those treated with bST. In mammary tissue, releasing factor increased fat synthesis from acetate more than did bST treatment, but percentages of fat or short-chain fatty acids in milk were not different between treatments, nor were differences significant between treatments and control for fatty acid esterification or lipoprotein lipase activity in mammary tissue or gluconeogenesis and fatty acid esterification in liver. We conclude that protein concentration was lower in omental adipose tissue of cows treated with releasing factor than that in tissue of cows treated with bST, although both treatments similarly suppressed synthesis of fat from acetate and activity of lipoprotein lipase.

Reduction of bovine plasma cholesterol concentration by partial interruption of enterohepatic circulation of bile salts: a novel hypocholesterolemic model.

Interruption of enterohepatic circulation (EHC) of bile salts in several species is known to cause a significant decrease in plasma concentrations of low density lipoprotein (LDL) cholesterol, but to have little effect on high density lipoprotein (HDL) cholesterol. The present study, for the first time, demonstrates that partial interruption of EHC dramatically reduces both plasma LDL and HDL cholesterol concentrations in cattle. Five adult Holstein cows were surgically altered to allow controlled portions of bile flow to be diverted from the body. The animals were fed a low-fat, cholesterol-free diet. In two experiments, bile was diverted at 50% and 22% of total flow rates. By day 8 of diversion, both rates reduced mean plasma cholesterol from baseline (85 mg/dl) to about 8 and 18 mg/dl, respectively. Cholesterol was reduced in equal proportions in all lipoprotein fractions. In addition, plasma concentrations of triglycerides and phospholipids were also dramatically reduced. All of these plasma lipids returned to baseline within 1 week after restoration of bile flow. To determine the hepatic response to bile diversion, liver cholesterol concentrations, cholesterol synthesis rates, and LDL receptor-binding activities were determined in biopsy samples. In response to bile diversion, hepatic cholesteryl esters were markedly depleted while hepatic cholesterol synthesis rates were increased by more than 10-fold. Nevertheless, because the basal cholesterol synthesis rate was so low, it was estimated that the increase in synthesis would have supplied no more than 5% of the sterols depleted during bile diversion (1.2 vs. 25 mmol/day). LDL receptor-binding activity was significantly elevated, suggesting an increased uptake of plasma lipoprotein cholesterol by the liver. These results suggest that the unique sensitivity of bovine plasma cholesterol to enterohepatic circulation interruption might occur as a result of the inherently low rate of hepatic cholesterol synthesis in cattle. This hypocholesterolemic model might serve as an interesting tool for the study of factors regulating plasma HDL cholesterol.

Metabolism of long chain fatty acids by ruminant liver.

The primary source of fatty acids processed by ruminant liver is nonesterified fatty acids (NEFA) from blood. Uptake is regulated by concentration of NEFA and blood flow. Blood NEFA concentration increases with negative energy balance. Blood flow increases with energy intake. Uptake and secretion of triacylglycerol between blood and the liver is limited. The reason for limited hepatic secretion of triacylglycerol-rich lipoprotein is unclear but probably involves the secretory process, not synthesis of triacylglycerol or apolipoprotein. Oxidation of fatty acids and ketogenesis are inhibited by malonyl-CoA and propionic acid. The stress of late gestation and early lactation increases NEFA supply to the liver, where they cause deposition of fat. Ketogenesis and oxidation in the liver increase but not sufficiently to prevent an accumulation of fat, which may contribute to decreased feed intake in the peripartum period.

A species comparison of liver slice synthesis and secretion of triacylglycerol from nonesterified fatty acids in media.

The ability of liver slices from eight species to synthesize and secrete triacylglycerol from nonesterified fatty acids contained in media was investigated. Species were grouped according to the relative proportion of lipogenesis occurring in the liver. The rate of liver triacylglycerol synthesis from nonesterified fatty acids in media was similar among species studied. Liver slices from species in which the liver contribution to lipogenesis is minor (sheep, cattle, pig and guinea pig) secreted less triacylglycerol synthesized from nonesterified fatty acids than did liver slices from species in which lipogenesis occurs predominantly in the liver (chicken and fish) or in liver and adipose tissue (rat and rabbit). The results suggest that the ability of liver to secrete triacylglycerol in very low density lipoproteins is proportional to the liver's lipogenic capacity.

Effects of dietary amines on small intestinal variables in neonatal pigs fed soy protein isolate.

Six litters of newborn crossbred piglets were utilized to examine 1) the effects of substituting 20% of the protein of an all-milk protein liquid diet with a soy protein isolate (milk-soy diet) on small intestinal variables and 2) the effects of supplementing this milk-soy diet with 25 g of either putrescine or ethylamine per kilogram diet on small intestinal variables. Small intestinal xylose absorption tended to increase from wk 1 to wk 2 of age in pigs fed the milk, putrescine and ethylamine diets, but not in pigs fed the milk, putrescine and ethylamine diets, but not in pigs fed the unsupplemented milk-soy diet. Crypt depth in pigs fed the milk-soy diet tended to be less (9.4%; P greater than .10) than the crypt depth in pigs fed the other diets, but mitotic index was not different (P greater than .10) among diets. Mucosal protein, DNA and RNA concentrations and mucosal brush border sucrase and cytosolic dipeptidase activities tended to be least in pigs fed the putrescine and ethylamine diets. Concentration of mucosal putrescine was greatest (P less than .002) in the distal regions of the small intestine of pigs fed putrescine. Mucosal ornithine decarboxylase activity was inhibited by putrescine (P less than .02), but it was not affected by the soybean protein isolate used in this study. Supplementing soy protein isolate diets with amines may enhance intestinal absorption and enterocyte proliferation.