Factors contributing to development of fatty liver and ketosis in lactating dairy cows

Interactions of feed restriction and an exogenous source of ketone bodies (1,3-butanediol) in causing fatty liver and ketosis in dairy cows were studied. In experiment one, 16 cows were assigned to either control (fed ad libitum), feed restriction (fed 80% of ad libitum), or ad libitum plus 5.5% butanediol from day 14 through day 42 postpartum. From day 43 through d 56 postpartum, cows previously in either feed-restricted or dietary butanediol groups received the two treatments combined. Treatments decreased milk production. Concentrations in plasma of nonesterified fatty acids, acetate and [beta]-hydroxybutyrate were increased 200, 35, and 115% during the first week of feed restriction. Insulin and 3-hydroxybutyrate were increased 13 and 50% in cows given dietary butanediol. Liver glycogen was 25% greater in cows given butanediol and was 38% lower in feed-restricted cows. Hepatic content of total lipid tended to be greater in cows given butanediol. Conversion of propionate to glucose by liver slices decreased during treatment for all groups and was decreased 15% by addition of L-carnitine to incubation media. Combining feed restriction and dietary butanediol at d 43 postpartum decreased plasma glucose and increased 3-hydroxybutyrate. Neither feed restriction nor butanediol as separate treatments, nor combining the two at d 43 postpartum, caused development of fatty liver or ketosis;In experiment two, 13 cows were assigned to either control (fed ad libitum) or ketosis induction (fed 80% of ad libitum plus 7% 1,3-butanediol) from day 14 through day 42 postpartum. Six cows in the ketosis induction group developed subclinical ketosis, whereas the other cow quickly developed clinical ketosis. Milk production was decreased 20% and milk fat content increased 11% by subclinical ketosis. Energy balance reached a low of -6.6 Mcal/d during the first week of ketosis induction. In subclinically ketotic cows, concentrations of nonesterified fatty acids and [beta]-hydroxybutyrate were increased over twofold. Dextran sulfate-precipitable cholesterol and triglyceride in serum increased during the first week of ketosis induction. Liver triglyceride and glycogen increased 200% and 50% in subclinically ketotic cows. Oxidation of palmitate was greater in liver slices from ketosis-induction cows. Ratios of triglyceride to glycogen greater than 2.0 in liver during the early-postpartum period may indicate susceptibility to clinical ketosis

Extruded Blend of Soybean Meal and Sunflower Seeds for Dairy Cattle in Early Lactation

An extruded blend of 44% crude protein soybean meal (50%), sunflower seeds (45%), and premix (5%) was evaluated as a protein and energy source for dairy cows in early lactation. Thirty Holstein cows (24 multiparos and 6 primiparous) were assigned to either a corn-oats-soybean meal concentrate (SBM) or a concentrate where soy-sunflower blend replaced all soybean meal and portions of corn and oats (SSF). Dry matter of total mixed diets was 36% corn silage, 21% alfalfa haylage, and 43% concentrate. Yields of milk (33.6 and 33.8 kg/day) and 4% fat-corrected milk (30.9 and 30.5 kg/day) were similar for cows fed SBM or SSF, while percentages of total solids (11.92 and 11.38), fat (3.55 and 3.30), protein (2.91 and 2.74), and solids-not-fat (8.38 and 8.09) were lower in milk from cows fed SSF. Milk from cows fed SSF contained fewer short and medium chain fatty acids, more 18-carbon fatty acids, and was more unsaturated than from cows fed SBM. Intakes of dry matter and changes in body weight were not different between diets. Ruminal fluid pH and molar ratio of acetate to propionate were higher in cows fed SSF, while concentrations of total volatile fatty acids and ammonia were lower in cows fed SSF. Concentrations of essential amino acids in arterial serum and calculated uptakes of amino acids by the mammary gland were similar between diets. Transfer efficiencies of phenylalanine, valine, isoleucine, leucine, and tyrosine were lower in cows fed SSF. In cows fed either diet, methionine appeared to be most limiting, while tryptophan and arginine were least limiting

Better postpartal performance in dairy cows supplemented with rumen-protected methionine compared with choline during the peripartal period

The onset of lactation in dairy cows is characterized by high output of methylated compounds in milk when sources of methyl group are in short supply. Methionine and choline (CHOL) are key methyl donors and their availability during this time may be limiting for milk production, hepatic lipid metabolism, and immune function. Supplementing rumen-protected Met and CHOL may improve overall performance and health of transition cows. The objective of this study was to evaluate the effect of supplemental rumen-protected Met and CHOL on performance and health of transition cows. Eighty-one multiparous Holstein cows were used in a randomized, complete, unbalanced block design with 2×2 factorial arrangement of Met (Smartamine M, Adisseo NA, Alpharetta, GA) and CHOL (ReaShure, Balchem Inc., New Hampton, NY) inclusion (with or without). Treatments (20 to 21 cows each) were control (CON), CON+Met (SMA), CON+CHOL (REA), and CON+Met+CHOL (MIX). From -50 to -21d before expected calving, all cows received the same diet (1.40Mcal of NEL/kg of DM) with no Met or CHOL. From -21d to calving, cows received the same close-up diet (1.52Mcal of NEL/kg of DM) and were assigned randomly to treatments (CON, SMA, REA, or MIX) supplied as top dresses. From calving to 30 DIM, cows were fed the same postpartal diet (1.71Mcal of NEL/kg of DM) and continued to receive the same treatments through 30 DIM. The Met supplementation was adjusted daily at 0.08% DM of diet and REA was supplemented at 60g/d. Incidence of clinical ketosis and retained placenta tended to be lower in Met-supplemented cows. Supplementation of Met (SMA, MIX) led to greater DMI compared with other treatments (CON, REA) in both close-up (14.3 vs. 13.2kg/d, SEM 0.3) and first 30d postpartum (19.2 vs. 17.2kg/d, SEM 0.6). Cows supplemented with Met (SMA, MIX) had greater yields of milk (44.2 vs. 40.4kg/d, SEM 1.2), ECM (44.6 vs. 40.5kg/d, SEM 1.0), and FCM (44.6 vs. 40.8kg/d, SEM 1.0) compared with other (CON, REA) treatments. Milk fat content did not differ in response to Met or CHOL. However, milk protein content was greater in Met-supplemented (3.32% vs. 3.14%, SEM 0.04%) but not CHOL-supplemented (3.27 vs. 3.19%, SEM 0.04%) cows. Supplemental CHOL led to greater blood glucose and insulin concentrations with lower glucose:insulin ratio. No Met or CHOL effects were detected for blood fatty acids or BHB, but a Met × time effect was observed for fatty acids due to higher concentrations on d 20. Results from the present study indicate that peripartal supplementation of rumen-protected Met but not CHOL has positive effects on cow performance

Biomarkers of inflammation, metabolism, and oxidative stress in blood, liver, and milk reveal a better immunometabolic status in peripartal cows supplemented with Smartamine M or MetaSmart

The peripartal dairy cow experiences a state of reduced liver function coupled with increased inflammation and oxidative stress. This study evaluated the effect of supplementing basal diets with rumen-protected Met in the form of MetaSmart (MS) or Smartamine M (SM) (both from Adisseo Inc., Antony, France) during the peripartal period on blood and hepatic biomarkers of liver function, inflammation, and oxidative stress. Thirty-seven multiparous Holstein cows were fed the same basal diet from −50 to −21 d relative to expected calving [1.24 Mcal/kg of dry matter (DM); no Met supplementation]. From −21 d to calving, the cows received diets (1.54 Mcal/kg of DM) with no added Met (control, CON; n = 13), CON plus MS (n = 11), or CON plus SM (n = 13). From calving through 30 d in milk (DIM), the cows received the same postpartal diet (1.75 Mcal/kg of DM; CON), or CON plus MS or CON plus SM. Liver and blood samples were harvested at various time points from −21 to 21 d relative to calving. Preplanned contrasts of CON versus SM + MS during prepartum (−21 and −10 d before calving) and postpartum (7, 14, and 21 d after calving) responses were evaluated. Cows fed MS or SM compared with CON had lower overall concentrations of plasma ceruloplasmin and serum amyloid A (SAA). Compared with CON, Met-supplemented cows had greater overall plasma oxygen radical absorbance capacity. Liver concentrations of glutathione and carnitine also were greater overall with Met supplementation. Milk choline and liver phosphatidylcholine were lower overall in cows fed Met compared with controls. Liver tissue choline concentrations did not differ. Data indicate that supplemental Met enhanced de novo glutathione and carnitine synthesis in liver and, thus, increased antioxidant and β-oxidation capacity. The greater decrease of IL-6 after calving coupled with lower ceruloplasmin and SAA in Met-supplemented cows indicated a reduction in proinflammatory signaling within liver. The lower hepatic phosphatidylcholine in Met-supplemented cows might have been associated with greater assembly or export of very low density lipoproteins. Overall, biomarker analyses in blood and tissue indicate that the beneficial effect of feeding SM and MS on postpartal cow performance is due in part to a better immunometabolic status

Effect of the level of maternal energy intake prepartum on immunometabolic markers, polymorphonuclear leukocyte function, and neutrophil gene network expression in neonatal Holstein heifer calves1

A conventional approach in dairy cow nutrition programs during late gestation is to feed moderate-energy diets. The effects of the maternal plane of nutrition on immune function and metabolism in newborn calves are largely unknown. Holstein cows (n=20) were fed a controlled-energy (CON) diet (1.24 Mcal/kg) for the entire dry period (~50 d) or the CON diet during the first 29 d of the dry period followed by a moderate-energy (OVE) diet (1.47 Mcal/kg) during the last 21 d prepartum. All calves were weighed at birth before first colostrum intake. Calves chosen for this study (n=6 per maternal diet) had blood samples harvested before colostrum feeding (d 0) and at 2 and 7 d of age. Blood samples were used to determine metabolites, acute-phase proteins, oxidative stress markers, hormones, phagocytic capacity of polymorphonuclear leukocytes (PMN) and monocytes, and total RNA was isolated from PMN. Calves from OVE dams weighed, on average, 5kg less at birth (44.0 vs. 48.6kg) than calves from CON dams. Blood glucose concentration in OVE calves had a more pronounced increase between 0 and 2 d than CON, at which point phagocytosis by PMN averaged 85% in OVE and 62% in CON. Compared with CON, calves from OVE had greater expression of TLR4, but lower expression of PPARA and PPARD at birth. Expression of PPARG and RXRA decreased between 0 and 2 d in both groups. Concentrations of leptin, cholesterol, ceruloplasmin, reactive oxygen metabolites, myeloperoxidase, retinol, tocopherol, IgG, and total protein, as well as expression of SOD2 and SELL increased markedly by 2 d in both groups; whereas, cortisol, albumin, acid-soluble protein, NEFA, insulin, as well as expression of IL6, TLR4, IL1R2, LTC4S, and ALOX5 decreased by 2 d. By 7 d of age, the concentration of haptoglobin was greater than precolostrum and was lower for OVE than CON calves. Our data provide evidence for a carry-over effect of maternal energy overfeeding during the last 3 wk before calving on some measurements of metabolism in the calf at birth and the phagocytic capacity of blood neutrophils after colostrum feeding. It might be feasible to design nutrient supplements to fortify colostrum in a way that metabolic and immunologic capabilities of the calf are improved

Liver lipid content and inflammometabolic indices in peripartal dairy cows are altered in response to prepartal energy intake and postpartal intramammary inflammatory challenge.

This study evaluated the effect of feeding a control diet (CON) or a moderate energy diet (overfed, OVE) during the dry period (∼45d) and a postpartum intramammary lipopolysaccharide (LPS) challenge on blood metabolic and inflammatory indices, milk production, and hepatic gene expression. A subset of cows (n=9/diet) in CON (1.34 Mcal/kg of dry matter) and OVE (1.62 Mcal/kg of dry matter) received an intramammary LPS challenge (200 μg; CON-LPS, OVE-LPS, respectively). Liver biopsies were harvested at -14 d from calving, and postpartum at 2.5h post-LPS on d 7, 14, and 30. Prepartum, the OVE group was in more positive energy balance (EB) and had greater body condition score compared with CON. In contrast, during wk 1 postpartum and before the LPS challenge, the OVE group was in greater negative EB than CON. Prepartal diet did not affect postpartal milk production or dry matter intake. At 2h postchallenge on d 7, we observed an increase in serum nonesterified fatty acids (NEFA) and bilirubin and a decrease in hydroxybutyrate, regardless of diet. That was coupled with greater haptoglobin in OVE-LPS compared with CON-LPS. In addition, OVE-LPS cows versus CON nonchallenged, OVE nonchallenged, and CON-LPS had greater liver triacylglycerol (TAG) concentration 2.5h postchallenge on d 7. The concentration of TAG in liver of OVE-LPS remained elevated by 30d postpartum. The liver TAG concentration in OVE-LPS compared with CON-LPS cows was associated with greater serum concentration of NEFA and reactive oxygen metabolites on d 10 and 14 postpartum. Cows in OVE-LPS also had greater concentrations of ceruloplasmin, cholesterol, and vitamin E from d 10 through 21. Among 28 genes associated with fatty acid oxidation, inflammation, oxidative stress, and gluconeogenesis, only SAA3 (which encodes an acute phase protein) was greater in CON-LPS compared with OVE-LPS at 2.5h postchallenge. Expression of HP, which encodes another acute phase protein, was greater in OVE-LPS than in CON-LPS at 14 and 30 d postpartum. Several inflammation-related genes (TNF, IRAK1, NFKB1, ANGPTL4) showed markedly decreased expression between 7 and 14 d, after which expression remained unchanged. No differences were observed in several genes of the growth-hormone/insulin-like growth factor-1 axis, except for SOCS2, expression of which decreased markedly between 7 and 14 d in OVE-LPS but not in CON-LPS. These data suggest that overfeeding a moderate-energy diet prepartum alters the response of the cow to an intramammary challenge after calving and may predispose it to sustained liver lipidosis

Blood immunometabolic indices and polymorphonuclear neutrophil function in peripartum dairy cows are altered by level of dietary energy prepartum

Cows experience some degree of negative energy balance and immunosuppression around parturition, making them vulnerable to metabolic and infectious diseases. The effect of prepartum feeding of diets to meet (control, 1.34 Mcal/kg of dry matter) or exceed (overfed, 1.62 Mcal/kg of dry matter) dietary energy requirements was evaluated during the entire dry period (∼45 d) on blood polymorphonuclear neutrophil function, blood metabolic and inflammatory indices, and milk production in Holstein cows. By design, dry matter intake in the overfed group (n=9) exceeded energy requirements during the prepartum period (-4 to -1 wk relative to parturition), resulting in greater energy balance when compared with the control group (n=10). Overfed cows were in more negative energy balance during wk 1 after calving than controls. No differences were observed in dry matter intake, milk yield, and milk composition between diets. Although nonesterified fatty acid concentration pre- (0.138 mEq/L) and postpartum (0.421 mEq/L) was not different between diets, blood insulin concentration was greater in overfed cows prepartum (16.7 μIU/mL) compared with controls pre- and postpartum (∼3.25 μIU/mL). Among metabolic indicators, concentrations of urea (4.63 vs. 6.38 mmol/L), creatinine (100 vs. 118 μmol/L), and triacylglycerol (4.0 vs. 8.57 mg/dL) in overfed cows were lower prepartum than controls. Glucose was greater pre- (4.24 vs. 4.00 mmol/L) and postpartum (3.49 vs. 3.30 mmol/L) compared with control cows. Among liver function indicators, the concentration of bilirubin increased by 2 to 6 fold postpartum in control and overfed cows. Phagocytosis capacity of polymorphonuclear neutrophils was lower prepartum in overfed cows (32.7% vs. 46.5%); phagocytosis in the control group remained constant postpartum (50%) but it increased at d 7 in the overfed group to levels similar to controls (48.4%). Regardless of prepartum diet, parturition was characterized by an increase in nonesterified fatty acid and liver triacylglycerol, as well as blood indices of inflammation (ceruloplasmin and haptoglobin), oxidative stress (reactive oxygen metabolites), and liver injury (glutamic oxaloacetic transaminase). Concentrations of the antioxidant and anti-inflammatory compounds vitamin A, vitamin E, and β-carotene decreased after calving. For vitamin A, the decrease was observed in overfed cows (47.3 vs. 27.5 μg/100 mL). Overall, overfeeding energy and higher energy status prepartum led to the surge of insulin and had a transient effect on metabolism postpartum

Gene network and pathway analysis of bovine mammary tissue challenged with Streptococcus uberis reveals induction of cell proliferation and inhibition of PPARγ signaling as potential mechanism for the negative relationships between immune response and lipid metabolism

<p>Abstract</p> <p>Background</p> <p>Information generated via microarrays might uncover interactions between the mammary gland and <it>Streptococcus uberis </it>(<b><it>S. uberis</it></b>) that could help identify control measures for the prevention and spread of <it>S. uberis </it>mastitis, as well as improve overall animal health and welfare, and decrease economic losses to dairy farmers. The main objective of this study was to determine the most affected gene networks and pathways in mammary tissue in response to an intramammary infection (<b>IMI</b>) with <it>S. uberis </it>and relate these with other physiological measurements associated with immune and/or metabolic responses to mastitis challenge with <it>S. uberis </it>O140J.</p> <p>Results</p> <p><it>Streptococcus uberis </it>IMI resulted in 2,102 (1,939 annotated) differentially expressed genes (<b>DEG</b>). Within this set of DEG, we uncovered 20 significantly enriched canonical pathways (with 20 to 61 genes each), the majority of which were signaling pathways. Among the most inhibited were <it>LXR/RXR Signaling </it>and <it>PPARα/RXRα Signaling</it>. Pathways activated by IMI were <it>IL-10 Signaling </it>and <it>IL-6 Signaling </it>which likely reflected counter mechanisms of mammary tissue to respond to infection. Of the 2,102 DEG, 1,082 were up-regulated during IMI and were primarily involved with the immune response, e.g., <it>IL6</it>, <it>TNF</it>, <it>IL8, IL10, SELL, LYZ</it>, and <it>SAA3</it>. Genes down-regulated (1,020) included those associated with milk fat synthesis, e.g., <it>LPIN1, LPL, CD36</it>, and <it>BTN1A1</it>. Network analysis of DEG indicated that <it>TNF </it>had positive relationships with genes involved with immune system function (e.g., <it>CD14, IL8, IL1B</it>, and <it>TLR2</it>) and negative relationships with genes involved with lipid metabolism (e.g., <it>GPAM</it>, <it>SCD</it>, <it>FABP4</it>, <it>CD36</it>, and <it>LPL</it>) and antioxidant activity (<it>SOD1</it>).</p> <p>Conclusion</p> <p>Results provided novel information into the early signaling and metabolic pathways in mammary tissue that are associated with the innate immune response to <it>S. uberis </it>infection. Our study indicated that IMI challenge with <it>S. uberis </it>(strain O140J) elicited a strong transcriptomic response, leading to potent activation of pro-inflammatory pathways that were associated with a marked inhibition of lipid synthesis, stress-activated kinase signaling cascades, and PPAR signaling (most likely PPARγ). This latter effect may provide a mechanistic explanation for the inverse relationship between immune response and milk fat synthesis.</p