Innate immunity and metabolomic responses in dairy cows challenged intramammarily with lipopolysaccharide after subacute ruminal acidosis

Subacute ruminal acidosis (SARA) is a prevalent metabolic disorder in dairy cows known to elicit local and systemic immune responses. We recently showed that cows experiencing SARA and challenged intramammarily with lipopolysaccharide (LPS) experienced stronger metabolic disturbances compared with cows without SARA. Therefore, we hypothesized that cows experiencing SARA have a modulated innate immune response and impaired plasma metabolome compared with healthy cows when experiencing an acute mastitis challenge. A total of 18 Simmental cows were subjected either to a Control (CON, n=6) or SARA (n=12) feeding regimen, receiving either 40% or 60% concentrates for 30 days. Thereafter, six SARA (SARA-LPS) and the CON (CON-LPS) cows were intramammarily challenged with 50 µg LPS from Escherichia coli (O26 : B6), while the remaining six SARA cows (SARA-PLA) received a placebo. Blood and milk samples were analyzed for acute phase proteins and a targeted ESI-LC-MS/MS-based metabolomics approach was performed in blood samples 24 h after the LPS challenge. The LPS infusion caused a strong increase in immune response variables, with a higher concentration of milk amyloid A 48 h after the LPS challenge in SARA-LPS compared with CON-LPS cows. Cows receiving the LPS infusion had a lower plasma concentration of several amino acids and lysophosphatidylcholines but without differences in SARA cows and healthy cows. In conclusion, our results revealed that an intramammary LPS infusion increased acute phase proteins and modulated the blood metabolome. While no systemic differences between SARA and healthy cows were observed, cows experiencing SARA showed a higher concentration of an acute phase protein at the local level of the mammary gland. Further research is required to elucidate the underlying mechanisms and to evaluate its clinical significance for udder health.     

Evaluation of the Change of Serum Copper and Zinc Concentrations of Dairy Cows with Subclinical Ketosis

Ketosis in dairy cows can lead to poor reproductive success and decreased milk production. Since the serum concentrations of copper (Cu) and zinc (Zn) are closely associated with the health status of cows, we investigated whether serum concentrations of Cu and Zn differed in dairy cows with subclinical ketosis and healthy dairy cows. Blood samples of 19 healthy dairy cows and 15 subclinically ketotic dairy cows were collected from three farms, and the concentrations of β-hydroxybutyrate (BHBA), glucose, non-esterified fatty acids (NEFA), Cu, and Zn were determined. Subclinically ketotic dairy cows had significantly higher BHBA and NEFA levels (p?<?0.01) and lower glucose (p?<?0.01) than healthy dairy cows. Likewise, serum concentrations of Zn were significantly decreased (p?<?0.05) in dairy cows with subclinical ketosis. There was no significant difference observed for serum Cu concentration between healthy and subclinically ketotic dairy cows. This study suggests that a decreased serum Zn concentration could be a cause of decreased reproductive performance in subclinically ketotic dairy cows.     

Effects of dietary energy source on energy balance, metabolites and reproduction variables in dairy cows in early lactation

This paper summarizes three recent studies by the same authors with the objective to study the effect of dietary energy source on the energy balance (EB) and risk for metabolic and reproductive disorders in dairy cows in early lactation. The first study, a literature survey, illustrated that feeding extra glucogenic nutrients relative to lipogenic nutrients, decreased milk fat and seems to decrease plasma non-esterified fatty acid (NEFA) and beta-hydroxybutyrate (BHBA) concentration. However studies are scarce and mostly confound the effect of energy source with level of energy intake, compromising eventual effects on EB and fertility. Therefore, in the second study, 16 dairy cows were either fed a glucogenic or a lipogenic diet (isocaloric) and EB was determined in climate-controlled respiration chambers from week 2 until 9 of lactation. Glucogenic diet decreased milk fat yield and milk energy and tended to decrease body fat mobilization compared with lipogenic diet. The objective of the third study was to study the effect of dietary energy source on EB, metabolites and reproduction variables. Dairy cows (n=111) were fed glucogenic, lipogenic or mixed diet from week 3 until week 9 relative to calving. Multiparous cows fed glucogenic diet had lower milk fat yield, higher calculated EB, and lower plasma NEFA, BHBA and liver triacylglyceride concentration and tended to have fewer days to first postpartum ovulation. In conclusion, increasing the glucogenic nutrient availability improved the EB and had potential to reduce the risk for metabolic disorders and to improve reproductive performance in dairy cows.     

Supplementation of a clay mineral-based product modulates plasma metabolomic profile and liver enzymes in cattle fed grain-rich diets

Grain-rich diets often lead to subacute ruminal acidosis (SARA) impairing rumen and systemic cattle health. Recent data suggest beneficial effects of a clay mineral (CM)- based product on the rumen microbiome of cattle during SARA. This study sought to investigate whether the CM supplementation can counteract SARA-induced perturbations of the bovine systemic health. The study used an intermittent diet-induced SARA-model with eight dry Holstein cows receiving either no additive as control or CM via concentrates (n=8 per treatment). Cows received first a forage diet (Baseline) for 1 week, followed by a 1-week SARA-challenge (SARA 1), a 1-week recovery phase (Recovery) and finally a second SARA-challenge for 2 weeks (SARA 2). Cows were monitored for feed intake, reticular pH and chewing behavior. Blood samples were taken and analyzed for metabolites related to glucose and lipid metabolism as well as liver health biomarkers. In addition, a targeted electrospray ionization-liquid chromatography-MS-based metabolomics approach was carried out on the plasma samples obtained at the end of the Baseline and SARA 1 phase. Data showed that supplementing the cows’ diet with CM improved ruminating chews per regurgitated bolus by 16% in SARA 1 (P=0.01) and enhanced the dry matter intake during the Recovery phase (P=0.05). Moreover, the SARA-induced decreases in several amino acids and phosphatidylcholines were less pronounced in cows receiving CM (P≤0.10). The CM-supplemented cows also had lower concentrations of lactate (P=0.03) and biogenic amines such as histamine and spermine (P<0.01) in the blood. In contrast, the concentration of acylcarnitines with key metabolic functions was increased in the blood of treated cows (P≤0.05). In SARA 2, the CM-cows had lower concentrations of the liver enzymes aspartate aminotransferase and γ-glutamyltransferase (P<0.05). In conclusion, the data suggest that supplementation of CM holds the potential to alleviate the negative effects of high-grain feeding in cattle by counteracting multiple SARA-induced perturbations in the systemic metabolism and liver health.     

Effects of trace mineral amount and source on aspects of oxidative metabolism and responses to intramammary lipopolysaccharide challenge in midlactation dairy cows

Trace minerals have important roles in immune function and oxidative metabolism; however, little is known about the relationships between supplementation level and source with outcomes in dairy cattle. Multiparous Holstein cows (n=48) beginning at 60 to 140 days in milk were utilized to determine the effects of trace mineral amount and source on aspects of oxidative metabolism and responses to intramammary lipopolysaccharide (LPS) challenge. Cows were fed a basal diet meeting National Research Council (NRC) requirements except for no added zinc (Zn), copper (Cu) or manganese (Mn). After a 4-week preliminary period, cows were assigned to one of four topdress treatments in a randomized complete block design with a 2×2 factorial arrangement of treatments: (1) NRC inorganic (NRC levels using inorganic (sulfate-based) trace mineral supplements only); (2) NRC organic (NRC levels using organic trace mineral supplements (metals chelated to 2-hydroxy-4-(methythio)-butanoic acid); (3) commercial inorganic (approximately 2×NRC levels using inorganic trace mineral supplements only; and (4) commercial organic (commercial levels using organic trace mineral supplements only). Cows were fed the respective mineral treatments for 6 weeks. Treatment effects were level, source and their interaction. Activities of super oxide dismutase and glutathione peroxidase in erythrocyte lysate and concentrations of thiobarbituric acid reactive substances (TBARS) and total antioxidant capacity (TAC) in plasma were measured as indices of oxidative metabolism. Effects of treatment on those indices were not significant when evaluated across the entire experimental period. Plasma immunoglobulin G level was higher in cows supplemented with organic trace minerals over the entire treatment period; responses assessed as differences of before and after Escherichia coli J5 bacterin vaccination at the end of week 2 of treatment period were not significant. Cows were administered an intramammary LPS challenge during week 5; during week 6 cows fed commercial levels of Zn, Cu and Mn tended to have higher plasma TAC and cows fed organic sources had decreased plasma TBARS. After the LPS challenge, the extent and pattern of response of plasma cortisol concentrations and clinical indices (rectal temperature and heart rate) were not affected by trace mineral level and source. Productive performance including dry matter intake and milk yield and composition were not affected by treatment. Overall, results suggest that the varying level and source of dietary trace minerals do not have significant short-term effects on oxidative metabolism indices and clinical responses to intramammary LPS challenge in midlactation cows.     

Transition Diseases in Grazing Dairy Cows Are Related to Serum Cholesterol and Other Analytes

The objectives of this study were to describe the incidence of postpartum disease and to evaluate the association with serum cholesterol concentrations during the first 3 weeks after calving in grazing dairy cows. The association between non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), calcium and postpartum diseases was also evaluated. A total of 307 Holstein dairy cows from 6 commercial grazing herds in Osorno, Chile, were monitored from calving until 21 days in milk. Cases of retained placenta, clinical hypocalcemia and clinical mastitis were recorded by the farmer using established definitions. Twice weekly, cows were evaluated for metritis by the same veterinarian based on vaginal discharge and body temperature. Postpartum blood samples were collected weekly and analyzed for serum concentrations of cholesterol, NEFA, BHBA and calcium. Cows were considered as having subclinical ketosis if BHBA >1.2 mmol/L, and subclinical hypocalcemia if calcium <2.0 mmol/L in any of the 3 weekly samples. Overall, 56% of the cows studied developed at least one clinical or subclinical disease after calving. Incidence of individual diseases was 8.8% for retained placenta, 4.2% for clinical hypocalcemia, 11.7% for clinical mastitis, 41.1% for metritis, 19.9% for subclinical hypocalcemia and 16.6% for subclinical ketosis. Lower postpartum cholesterol in cows was associated with developing severe metritis or having more than one clinical disease after calving. For every 0.4 mmol/L decrease in serum cholesterol cows were nearly twice as likely to be diagnosed with multiple clinical diseases after calving. Higher BHBA concentrations and lower calcium concentrations during week 1 were associated with severe cases of metritis. Low serum calcium concentration during week 1 was also associated with developing more than one clinical disorder after calving. In conclusion, the incidence of postpartum diseases can be high even in grazing herds and lower serum cholesterol concentrations were associated with occurrence of clinical postpatum disorders.     

Diet-induced bacterial immunogens in the gastrointestinal tract of dairy cows: Impacts on immunity and metabolism

Dairy cows are often fed high grain diets to meet the energy demand for high milk production or simply due to a lack of forages at times. As a result, ruminal acidosis, especially subacute ruminal acidosis (SARA), occurs frequently in practical dairy production. When SARA occurs, bacterial endotoxin (or lipopolysaccharide, LPS) is released in the rumen and the large intestine in a large amount. Many other bacterial immunogens may also be released in the digestive tract following feeding dairy cows diets containing high proportions of grain. LPS can be translocated into the bloodstream across the epithelium of the digestive tract, especially the lower tract, due to possible alterations of permeability and injuries of the epithelial tissue. As a result, the concentration of blood LPS increases. Immune responses are subsequently caused by circulating LPS, and the systemic effects include increases in concentrations of neutrophils and the acute phase proteins such as serum amyloid-A (SAA), haptoglobin (Hp), LPS binding protein (LBP), and C-reactive protein (CRP) in blood. Entry of LPS into blood can also result in metabolic alterations. Blood glucose and nonesterified fatty acid concentrations are enhanced accompanying an increase of blood LPS after increasing the amount of grain in the diet, which adversely affects feed intake of dairy cows. As the proportions of grain in the diet increase, patterns of plasma β-hydoxybutyric acid, cholesterol, and minerals (Ca, Fe, and Zn) are also perturbed. The bacterial immunogens can also lead to reduced supply of nutrients for synthesis of milk components and depressed functions of the epithelial cells in the mammary gland. The immune responses and metabolic alterations caused by circulating bacterial immunogens will exert an effect on milk production. It has been demonstrated that increases in concentrations of ruminal LPS and plasma acute phase proteins (CRP, SAA, and LBP) are associated with declines in milk fat content, milk fat yield, 3.5% fat-corrected milk yield, as well as milk energy efficiency.     

Effects of continuous milking during a field trial on productivity,milk protein yield and health in dairy cows

The objective of this field study with an automatic milking system was to evaluate the effects of omitting the dry period on health and productivity during the subsequent lactation in dairy cows. A total of 98 German Simmental cows of six Southern German farms were assigned randomly to two experimental groups: The first group was dried-off 56 days before calving (D for dried-off, n=49), and the second group was milked continuously during this period until calving (CM for continuous milking, n=49). From the latter a third group emerged, including cows that dried-off themselves spontaneously (DS for dried-off spontaneously, n=14). Blood serum values of glucose, β-hydroxybutyrate (BHBA), non-esterified fatty acids (NEFA) and IGF-1 showed most pronounced fluctuations in D cows. Over the entire study period, the concentrations of BHBA and NEFA were markedly lower in the CM and DS groups. Furthermore, IGF-1 concentration was lowest for D cows and also decrease in back fat thickness was more pronounced. Mean concentration of milk protein was markedly higher in CM and DS cows (3.70% and 3.71%) compared with D cows (3.38%). Owing to the lower 305-day milk yield (−15.6%) and the lower total milk yield (−3.1%), the total amount of produced protein in the subsequent lactation was 2.5% (6.8 kg) lower, although the additional protein amount in CM cows from week −8 to calving was 35.7 kg. The greatest benefit resulted from positive effects on fertility and the lower incidence of diseases: CM cows had their first oestrus 1 week earlier compared with D cows, they also conceived earlier and showed a significantly lower risk of developing hypocalcaemia, ketosis and puerperal disorders. The present study showed that the costs of medical treatment and milk losses were twice as high in D cows, compared with CM and DS cows, and thus the reduced costs because of the more stable health outweighed the financial losses of milk yield by +18.49 € per cow and lactation.     

Metabolic adaptation and reticuloruminal pH in periparturient dairy cows experiencing different lipolysis early postpartum

Metabolic adaptation includes an array of concerted metabolic and endocrine events that enable dairy cows bridging the period of energy deficit at the onset of lactation. The present study evaluated metabolic, endocrine and reticuloruminal pH changes in 30 (25 Holstein and five Simmental) periparturient dairy cows experiencing variable lipolysis early postpartum. Cows were fed the same close-up and fresh lactation diets and kept in the same management conditions. Blood samples were collected at day 14, and day 4, relative to expected parturition, and at day 2, and day 21 postpartum, and serum metabolites and hormones related to glucose and lipid metabolism, as well as concentrations of several liver enzymes and acute phase proteins were determined. Additionally, reticuloruminal pH was monitored every 10 min over the last 3 days of the observation period. BW and milk yields were recorded and balances of energy and protein were assessed. Based on serum concentration of non-esterified fatty acids (NEFA) postpartum, cows were retrospectively classified into low (n=8), medium (n=11), and high (n=11) lipolysis groups, with NEFA levels of <0.4 mmol/l, between 0.4 and 0.7 mmol/l, and >0.7 mmol/l, respectively. Overall, elevated NEFA concentrations in the High group went along with a higher ratio of NEFA to cholesterol and reduced insulin sensitivity. While serum glucose, energy deficit and BW loss did not differ, cows of the High group exhibited increased lactate concentrations in the serum, compared with the Medium group. No differences in liver enzymes and acute phase proteins were evidenced among fat mobilization groups, whereas concentration of serum billirubin was lowest in the Low group after parturition. Data of milk yield and milk energy output showed no differences among groups, despite divergences in calculated energy balance and BW change postpartum. Cows of the Low group tended to increase dry matter intake but also showed longer time duration of pH below 6.0 in the reticulorumen (on average 299 min/day compared with 99 and 91 min/day for Medium and High groups, respectively). Differences in metabolic, endocrine and reticuloruminal pH responses indicate diverse metabolic adaptation strategies of early-lactation cows to cope with energy deficit postpartum.     

Transient reductions in milk fat synthesis and their association with the ruminal and metabolic profile in dairy cows fed high-starch,low-fat diets

Sub-acute ruminal acidosis (SARA) is sometimes observed along with reduced milk fat synthesis. Inconsistent responses may be explained by dietary fat levels. Twelve ruminally cannulated cows were used in a Latin square design investigating the timing of metabolic and milk fat changes during Induction and Recovery from SARA by altering starch levels in low-fat diets. Treatments were (1) SARA Induction, (2) Recovery and (3) Control. Sub-acute ruminal acidosis was induced by feeding a diet containing 29.4% starch, 24.0% NDF and 2.8% fatty acids (FAs), whereas the Recovery and Control diets contained 19.9% starch, 31.0% NDF and 2.6% FA. Relative to Control, DM intake (DMI) and milk yield were higher in SARA from days 14 to 21 and from days 10 to 21, respectively (P < 0.05). Milk fat content was reduced from days 3 to 14 in SARA (P < 0.05) compared with Control, while greater protein and lactose contents were observed from days 14 to 21 and 3 to 21, respectively (P < 0.05). Milk fat yield was reduced by SARA on day 3 (P < 0.05), whereas both protein and lactose yields were higher on days 14 and 21 (P < 0.05). The ruminal acetate-to-propionate ratio was lower, and the concentrations of propionate and lactate were higher in the SARA treatment compared with Control on day 21 (P < 0.05). Plasma insulin increased during SARA, whereas plasma non-esterified fatty acids and milk β-hydroxybutyrate decreased (P < 0.05). Similarly to fat yield, the yield of milk preformed FA (>16C) was lower on day 3 (P < 0.05) and tended to be lower on day 7 in SARA cows (P < 0.10), whereas yield of de novo FA (<16C) was higher on day 21 (P < 0.01) in the SARA group relative to Control. The t10- to t11-18:1 ratio increased during the SARA Induction period (P < 0.05), but the concentration of t10-18:1 remained below 0.5% of milk fat, and t10,c12 conjugated linoleic acid remained below detection levels. Odd-chain FA increased, whereas branched-chain FA was reduced during SARA Induction from days 3 to 21 (P < 0.05). Sub-acute ruminal acidosis reduced milk fat synthesis transiently. Such reduction was not associated with ruminal biohydrogenation intermediates but rather with a transient reduction in supply of preformed FA. Subsequent rescue of milk fat synthesis may be associated with higher availability of substrates due to increased DMI during SARA.     

Lipopolysaccharide markedly changes glucose metabolism and mitochondrial function in the longissimus muscle of pigs

Most previous studies on the effects of lipopolysaccharide (LPS) in pigs focused on the body’s immune response, and few reports paid attention to body metabolism changes. To better understand the glucose metabolism changes in skeletal muscle following LPS challenge and to clarify the possible mechanism, 12 growing pigs were employed. Animals were treated with either 2 ml of saline or 15 µg/kg BW LPS, and samples were collected 6 h later. The glycolysis status and mitochondrial function in the longissimus dorsi (LD) muscle of pigs were analyzed. The results showed that serum lactate content and NADH content in LD muscle significantly increased compared with the control group. Most glycolysis-related genes expression, as well as hexokinase, pyruvate kinase and lactic dehydrogenase activity, in LD muscle was significantly higher compared with the control group. Mitochondrial complexes I and IV significantly increased, while mitochondrial ATP concentration markedly decreased. Significantly increased calcium content in the mitochondria was observed, and endoplasm reticulum (ER) stress has been demonstrated in the present study. The results showed that LPS treatment markedly changes glucose metabolism and mitochondrial function in the LD muscle of pigs, and increased calcium content induced by ER stress was possibly involved. The results provide new clues for clarifying metabolic diseases in muscle induced by LPS.     

Relationship between metabolic status and behavior in dairy cows in week 4 of lactation

Blood metabolite and hormone concentrations are indicative of metabolic status, but blood sampling and analysis is invasive and time-consuming. Monitoring behavior can be done automatically, and behaviors may also be used as indicators of metabolic status. The aim of this study was to analyze the relationships between metabolic status and feeding behavior, lying behavior, motion index and steps of dairy cows in week 4 postpartum. Behavioral data from 81 Holstein-Friesian cows were collected using computerized feeders and accelerometers, and blood samples were collected for analysis of free-fatty acid (FFA), β-hydroxybutyrate (BHB), glucose, insulin, IGF-1 and growth hormone (GH) concentrations. First, cluster analysis was performed to categorize cows as having poor, average, good or very good metabolic status based on their plasma FFA, BHB, glucose, insulin, IGF-1 and GH concentration. Subsequently, the performance and behavior of cows in clusters with poor, average and good metabolic status were compared using GLM. Cows with a poor or average metabolic status tended to have greater fat-and-protein-corrected milk yield than cows with good metabolic status. Furthermore, cows with a poor metabolic status had a lower energy balance and dry matter intake (DMI) than cows with an average or good metabolic status and had a lower number of meals than cows with good metabolic status. Daily number of visits to the feeder and lying time tended to be positively related with metabolic status. Feeding rate (kg/min), daily meal time (min/day), number of lying bouts per day, steps and motion index were not related with metabolic status. In conclusion, better metabolic status in dairy cows in early lactation was associated with a greater DMI, increased feeding activity and a tendency to more time spent lying, compared with poor metabolic status. These results suggest that compromised metabolic status is reflected in altered cow’s behavior in week 4 of lactation.     

Relationships between thyroid hormones and serum energy metabolites with different patterns of postpartum luteal activity in high-producing dairy cows

This study investigated the relationships of thyroid hormones, serum energy metabolites, reproductive parameters, milk yield and body condition score with the different patterns of postpartum luteal activity in the postpartum period. A total of 75 multiparous healthy (free of detectable reproductive disorders) Holstein dairy cows (mean peak milk yield = 56.5 ± 7.0 kg/day) were used in this study. Transrectal ultrasound scanning and blood sample collection were performed twice weekly. Serum concentrations of progesterone (P₄) were measured twice weekly and beta-hydroxybutyrate (BHBA), non-esterified fatty acids, thyroxine (T₄), 3,30,5-tri-iodothyronine (T₃), free thyroxine (fT₄) and free 3,30,5-tri-iodothyronine (fT₃) were measured every 2 weeks from the 1st to the 8th week postpartum. On the basis of the serum P₄ profile of the cows, 25 (33.4%) had normal luteal activity (NLA), whereas 30 (40%), 10 (13.3%), 6 (8%) and 4 (5.3%) had prolonged luteal phase (PLP), delayed first ovulation (DOV), anovulation (AOV) and short luteal phase, respectively. Serum T₄ concentrations in PLP cows were higher than that in NLA cows at the 3rd week postpartum and did not change during the period of study, whereas in the NLA cows the concentrations increased (P < 0.05). Further, the least square (LS) mean of serum fT₄ concentrations in the DOV and AOV cows were significantly lower than in the NLA cows during the study period (P < 0.05). In addition, the AOV cows had higher LS mean serum BHBA and T₄ concentrations than the NLA cows in early weeks postpartum (P < 0.05). In conclusion, the serum thyroid hormones’ profile differs in high-producing dairy cows showing PLP, AOV and DOV in comparison with the postpartum NLA cows.     

Insulin Sensitivity in Adipose and Skeletal Muscle Tissue of Dairy Cows in Response to Dietary Energy Level and 2,4-Thiazolidinedione (TZD)

The effects of dietary energy level and 2,4-thiazolidinedione (TZD) injection on feed intake, body fatness, blood biomarkers and TZD concentrations, genes related to insulin sensitivity in adipose tissue (AT) and skeletal muscle, and peroxisome proliferator-activated receptor gamma (PPARG) protein in subcutaneous AT (SAT) were evaluated in Holstein cows. Fourteen nonpregnant nonlactating cows were fed a control low-energy (CON, 1.30 Mcal/kg) diet to meet 100% of estimated nutrient requirements for 3 weeks, after which half of the cows were assigned to a higher-energy diet (OVE, 1.60 Mcal/kg) and half of the cows continued on CON for 6 weeks. All cows received an intravenous injection of TZD starting 2 weeks after initiation of dietary treatments and for an additional 2 weeks, which served as the washout period. Cows fed OVE had greater energy intake and body mass than CON, and TZD had no effect during the administration period. The OVE cows had greater TZD clearance rate than CON cows. The lower concentration of nonesterified fatty acids (NEFA) and greater concentration of insulin in blood of OVE cows before TZD injection indicated positive energy balance and higher insulin sensitivity. Administration of TZD increased blood concentrations of glucose, insulin, and beta-hydroxybutyrate (BHBA) at 2 to 4 weeks after diet initiation, while the concentration of NEFA and adiponectin (ADIPOQ) remained unchanged during TZD. The TZD upregulated the mRNA expression of PPARG and its targets FASN and SREBF1 in SAT, but also SUMO1 and UBC9 which encode sumoylation proteins known to down-regulate PPARG expression and curtail adipogenesis. Therefore, a post-translational response to control PPARG gene expression in SAT could be a counteregulatory mechanism to restrain adipogenesis. The OVE cows had greater expression of the insulin sensitivity-related genes IRS1, SLC2A4, INSR, SCD, INSIG1, DGAT2, and ADIPOQ in SAT. In skeletal muscle, where PPARA and its targets orchestrate carbohydrate metabolism and fatty acid oxidation, the OVE cows had greater glyceroneogenesis (higher mRNA expression of PC and PCK1), whereas CON cows had greater glucose transport (SLC2A4). Administration of TZD increased triacylglycerol concentration and altered expression of carbohydrate- and fatty acid oxidation-related genes in skeletal muscle. Results indicate that overfeeding did not affect insulin sensitivity in nonpregnant, nonlactating dairy cows. The bovine PPARG receptor appears TZD-responsive, with its activation potentially leading to greater adipogenesis and lipogenesis in SAT, while differentially regulating glucose homeostasis and fatty acid oxidation in skeletal muscle. Targeting PPARG via dietary nutraceuticals while avoiding excessive fat deposition might improve insulin sensitivity in dairy cows during times such as the peripartal period when the onset of lactation naturally decreases systemic insulin release and sensitivity in tissues such as AT.     

Differential abundance of IGF1, bile acids,and the genes involved in their signaling in the dominant follicle microenvironment of lactating cows and nulliparous heifers

It is well documented that incidence of fertility problems is high in lactating cows but not in heifers of the same genetic merit. Understanding the metabolic and molecular differences between fertile heifers and relatively infertile lactating cows will help us understand the pathogenesis of infertility in dairy cows. Follicular waves in lactating cows (30–50 days in milk; n = 12) and heifers (n = 10) were synchronized by ultrasound-guided follicle ablation. Follicular fluid and granulosa cells of the dominant follicle were collected by ultrasound-guided aspiration along with blood sampling on Day 6 after synchronization. Dominant and subordinate follicles were larger in lactating cows than in heifers. Metabolic stress in lactating cows was evidenced by lower glucose and higher ß-hydroxy butyric acid compared with heifers. Insulin-like growth factor 1 signaling was reduced in the dominant follicle in lactating cows through reduced insulin-like growth factor 1 concentrations in plasma and follicular fluid of the dominant follicle, and reduced expression of pregnancy-associated plasma protein A (PAPPA) in their granulosa cells. We also found increased levels of total bile acids in the follicular fluid of the dominant follicle of lactating cows compared with heifers. Granulosa cells of the dominant follicle had higher expression of SLC10A2 and GPBAR1 (bile acid transporter and receptor, respectively) in lactating cows. These novel data are indicative of increased bile acid signaling within the dominant follicles of lactating cows compared with heifers. Overall, we demonstrate in the present study the metabolic, endocrine, and molecular differences within the microenvironment of the dominant follicles in lactating cows and heifers. These differences in follicular microenvironment may contribute toward abnormal ovarian function in lactating dairy cows.     

Feeding corn germ instead of corn grain on the performance of Holstein dairy cows fed low-forage diet and human-edible feed conversion efficiency

Using corn germ (CG) instead of corn grain could maintain dairy cow performance and might increase the efficiency of human food production. The primary objective of this study was to evaluate the effects of replacing corn grain with CG on the performance, nutrient intake, and digestibility of dairy cows. It also aimed to investigate the effect of CG on the efficiency of human food production in high-producing Holstein dairy cows in early lactation. Nine multiparous Holstein cows with 65.6 ± 8.5 DIM, milk yield of 55.6 ± 4.5 kg/d, and body weight of 611.3 ± 43.3 kg (mean ± SD) were used in a 3 × 3 Latin square design with 21-d periods. Treatments were (1) control treatment (CT, diet contains corn grain), (2) alternative treatment (AT, diet where corn grain was replaced with CG), and (3) balanced treatment (BT, diet where corn grain was replaced with CG but with the same energy content as CT). Control and balanced diets were isoenergetic (6.61 MJ/kg of DM); however, AT had higher energy (6.77 MJ/kg of DM). Treatments had no effect on dry and organic matter intake. NDF intake, however, was higher in CG diets compared with CT (P = 0.0001). Total-tract digestibility of DM tended to be reduced (P = 0.08), and OM digestibility was reduced (P = 0.05) by the inclusion of CG in diets. Whole and energy-corrected milk production were greater in AT compared with CT and BT (P < 0.05). Milk yield was similar in cows fed CT and BT. Treatments had no effect on milk composition or feed efficiency. Diet CT, when compared with CG diets, had lower efficiency in terms of human-edible feed conversion efficiency (HeFCE) and net food production (P < 0.05). Diet BT had greater HeFCE and net production of human-edible CP than AT (P < 0.05). Plasma BHBA, non-esterified fatty acids, and glucose concentrations were not affected by treatments, but plasma cholesterol was higher in cows that consumed CG diets (P = 0.04). The results indicate that, in high-producing early lactation dairy cows fed high concentrate diets, net food protein production can be substantially improved without lowering milk production through the reduction of dietary starch from 30.2 to 24.8% by replacing corn grain with CG.     

Physiological adaptations and ovarian cyclicity of Holstein and Montbéliarde cows under two low-input production systems

The objective was to study milk production, body reserve mobilization, metabolic and hormonal profiles, and ovarian cyclicity of Holstein-Friesian (HOLS) and Montbéliarde (MONT) cows under two low-input dairy production systems with seasonal spring calving: an extensive (EXT; 12 HOLS and 12 MONT) based on permanent diversified grasslands and zero concentrate, and a semi-extensive (SEMI; 12 HOLS and 10 MONT) based on established temporary grasslands and up to 4 kg/day of concentrate. Individual measurements were performed between −4 and 12 weeks of lactation. Cows in EXT secreted less milk (22.1 v. 24.4 kg/day), protein (660 v. 755 g/day) and energy (67.7 v. 74.4 MJ/day), had greater plasma β-hydroxybutyrate (BHBA) (0.97 v. 0.69 mM), lower glucose (59.0 v. 62.0 mg/dl) and IGF-1 (62 v. 71 ng/ml), lower milk fat concentration in fatty acids originating from de novo synthesis (e.g. ∑ 10:0 to 15:0) and greater concentration of those derived in part from mobilization of fat reserves (e.g. 18:0 and ∑>C16), and showed greater frequency of abnormal ovarian cycles compared with SEMI. Across production systems, HOLS produced more milk (24.7 v. 21.8 kg/day), protein (738 v. 674 g/day) and fat (939 v. 819 g/day), secreted more energy (75.1 v. 67.0 MJ/day), lost more body condition score (BCS) (1.41 v. 1.03) and reached a lower BCS nadir (1.12 v. 1.43), had greater plasma BHBA (0.91 v. 0.75 mM), lower insulin (15.9 v. 17.2 µIU/ml) and tended to have lower glucose (59.6 v. 61.4 mg/dl), had lower milk fat concentration in ∑ 10:0 to 15:0, tended to have higher ∑>C16 and tended to show more abnormal estrous cycles compared with MONT. Ultrasound measurements did not differentiate fat mobilization and were confounded by breed differences of skin thickness. The greater nutrient allowance in SEMI improved indicators of physiological status and ovarian function during early lactation compared with EXT, but did not attenuate body reserve mobilization because cows prioritized milk secretion. HOLS secreted more nutrients than MONT but lost more BCS, which negatively affected nutritional balance and tended to affect ovarian cyclicity during early lactation. Breed by system interactions were not observed except for a few variables.     

Changes in Serum Copper and Zinc Levels in Peripartum Healthy and Subclinically Hypocalcemic Dairy Cows

The objective of this study was to determine the levels of serum copper and zinc in subclinically hypocalcemic peripartum dairy cows in comparison to healthy animals. Blood samples were taken from 219 multiparous Holstein cows near parturition (from 4 weeks prepartum to 4 weeks postpartum) and 51 cows with subclinical hypocalcemia. The results showed that the serum copper concentration increased gradually at 1 week prepartum and remained high for the first 4 weeks postpartum in the healthy periparturient dairy cows. The serum zinc concentration reached a nadir at 1 week postpartum and subsequently increased gradually to baseline. The serum zinc concentration was significantly decreased (P?<?0.01) in dairy cows with subclinical hypocalcemia compared with healthy cows. There was no significant difference in the serum copper concentration between cows with subclinical hypocalcemia and healthy cows. These data demonstrate that the concentrations of copper and zinc in serum change dramatically during the peripartum period in dairy cows, which is a tremendous challenge for the body and for the maintenance of dairy cow health. The present study further suggests that a decreased serum zinc concentration could be a cause of decreased productive performance and increased susceptibility to other diseases due to immunosuppression in dairy cows with subclinical hypocalcemia. Additionally, this decreased zinc concentration may be involved in the pathogenesis of subclinical hypocalcemia.     

Rumen Microbiome Composition Determined Using Two Nutritional Models of Subacute Ruminal Acidosis

Subacute ruminal acidosis (SARA) is a metabolic disease in dairy cattle that occurs during early and mid-lactation and has traditionally been characterized by low rumen pH, but lactic acid does not accumulate as in acute lactic acid acidosis. It is hypothesized that factors such as increased gut permeability, bacterial lipopolysaccharides, and inflammatory responses may have a role in the etiology of SARA. However, little is known about the nature of the rumen microbiome during SARA. In this study, we analyzed the microbiome of 64 rumen samples taken from eight lactating Holstein dairy cattle using terminal restriction fragment length polymorphisms (TRFLP) of 16S rRNA genes and real-time PCR. We used rumen samples from two published experiments in which SARA had been induced with either grain or alfalfa pellets. The results of TRFLP analysis indicated that the most predominant shift during SARA was a decline in gram-negative Bacteroidetes organisms. However, the proportion of Bacteroidetes organisms was greater in alfalfa pellet-induced SARA than in mild or severe grain-induced SARA (35.4% versus 26.0% and 16.6%, respectively). This shift was also evident from the real-time PCR data for Prevotella albensis, Prevotella brevis, and Prevotella ruminicola, which are members of the Bacteroidetes. The real-time PCR data also indicated that severe grain-induced SARA was dominated by Streptococcus bovis and Escherichia coli, whereas mild grain-induced SARA was dominated by Megasphaera elsdenii and alfalfa pellet-induced SARA was dominated by P. albensis. Using discriminant analysis, the severity of SARA and degree of inflammation were highly correlated with the abundance of E. coli and not with lipopolysaccharide in the rumen. We thus suspect that E. coli may be a contributing factor in disease onset.The bovine rumen is a classical host-microbe symbiotic system, and disturbances in this exquisitely balanced ecosystem may lead to disease in the host. An example is subacute ruminal acidosis (SARA), or non-lactic acid acidosis, which has a disease etiology distinct from that of acute lactic acid acidosis because there is no accumulation of lactic acid (35). Field studies in the United States estimated that 19% of early lactating cows and 26% of mid-lactation cows suffered from SARA (11). In Germany and The Netherlands, approximately 11% of early lactation and 18% of mid-lactation cows suffered from this disease (22). In the acute form, lactic acid accumulates in the rumen, causing metabolic acidosis, and it usually occurs when animals are abruptly transitioned to a high-grain diet from a predominantly forage diet (38). If, however, the adaptation is gradual, slower-growing lactic acid-consuming bacteria, like Megasphaera elsdenii, convert the lactic acid to propionic acid (29). In SARA, lactic acid does not accumulate during low-pH conditions and other factors, like microbial population shifts and immune responses, appear to be associated with the disease etiology (35).In both acute and subacute acidosis, there is an increase in lipopolysaccharide (LPS) concentrations in the rumen (8, 14, 16). LPS and/or the low-pH rumen conditions may increase the permeability of the gut to LPS, which could trigger systemic inflammation (4). We previously developed two animal models of SARA, one based on grain and one based on alfalfa pellets (20, 21). Even though both models resulted in substantial reductions in rumen pH and an accumulation of LPS, only the grain induction model resulted in inflammation and the appearance of LPS in the peripheral blood (20, 21).In contrast to the rumen microbiome during lactic acid acidosis, the rumen microbiome during SARA has not been evaluated (13, 28). Even in acute acidosis, studies are largely culture based, and the uncultured members of the community have not been extensively assessed (31, 46, 49). In this article, we describe the rumen microbiome when two SARA induction models were used. The shifts in microbial community structure were assessed using terminal restriction fragment length polymorphism (TRFLP) analysis and real-time PCR of key microbial populations.