BHB Concentrations Research Articles

β-Hydroxybutyrate induces bovine hepatocyte apoptosis via an ROS-p38 signaling pathway.

β-Hydroxybutyrate (BHB) is an important indicator for metabolic disorders in dairy cows, such as ketosis and fatty liver. Dairy cows with ketosis display oxidative stress that may be associated with high levels of BHB. The purpose of this study was to demonstrate a correlation between the high levels of BHB and oxidative stress in dairy cows with ketosis, and to investigate the molecular mechanisms underlying oxidative damage in bovine hepatocytes. The results showed that dairy cows with ketosis exhibited oxidative stress and liver damage, which was significantly correlated with plasma BHB. Similarly, high concentrations of BHB increased the oxidative stress of cow hepatocytes in vitro, resulting in the phosphorylation and activation of p38 mitogen-activated protein kinase (MAPK), which led to increased expression, nuclear localization, and transcriptional activity of p53 and decreased Nrf2 in bovine hepatocytes. High concentrations of BHB significantly increased the expression of proapoptotic genes and significantly inhibited the expression of antiapoptotic genes. Finally, high concentrations of BHB promoted apoptosis in bovine hepatocytes. N-Acetyl-l-cysteine, glucose, and SB203580 (p38 inhibitor) significantly attenuated BHB-induced apoptotic damage in hepatocytes. These results indicate that BHB induces bovine hepatocyte apoptosis through the ROS-p38-p53/Nrf2 signaling pathway.

Effect of reducing milk production using a prolactin-release inhibitor or a glucocorticoid on metabolism and immune functions in cows subjected to acute nutritional stress.

When cows are unable to consume enough feed to support milk production, they often fall into severe negative energy balance. This leads to a weakened immune system and increases their susceptibility to infectious diseases. Reducing the milk production of cows subjected to acute nutritional stress decreases their energy deficit. The aim of this study was to compare the effects on metabolism and immune function of reducing milk production using quinagolide (a prolactin-release inhibitor) or dexamethasone in feed-restricted cows. A total of 23 cows in early/mid-lactation were fed for 5 d at 55.9% of their previous dry matter intake to subject them to acute nutritional stress. After 1 d of feed restriction and for 4 d afterward (d 2 to 5), cows received twice-daily i.m. injections of water (control group; n=8), 2mg of quinagolide (QN group; n=7), or water after a first injection of 20mg of dexamethasone (DEX group; n=8). Feed restriction decreased milk production, but the decrease was greater in the QN and DEX cows than in the control cows on d 2 and 3. As expected, feed restriction reduced the energy balance, but the reduction was lower in the QN cows than in the control cows. Feed restriction decreased plasma glucose concentration and increased plasma nonesterified fatty acid (NEFA) and β-hydroxybutyrate (BHB) concentrations. The QN cows had higher glucose concentration and lower BHB concentration than the control cows. The NEFA concentration was also lower in the QN cows than in the control cows on d 2. Dexamethasone injection induced transient hyperglycemia concomitant with a reduction in milk lactose concentration; it also decreased BHB concentration and decreased NEFA initially but increased it later. Feed restriction and quinagolide injections did not affect the blood concentration or activity of polymorphonuclear leukocytes (PMN), whereas dexamethasone injection increased PMN blood concentration but decreased the proportion of PMN capable of inducing oxidative burst. Incubation of peripheral blood mononuclear cells in serum harvested on d 2 of the restriction period reduced their ability to react to mitogen-induced proliferation, and injection of quinagolide or dexamethasone could not alleviate this effect. This experiment shows that prolactin-release inhibition could be an alternative to dexamethasone for reducing milk production and energy deficit in cows under acute nutritional stress, without disturbing immune function.

Milk yield, milk composition, and hepatic lipid metabolism in transition dairy cows fed flaxseed or linola.

The response of transition dairy cows to dietary supplementation with fat sources of various fatty acid profiles could affect hepatic fat metabolism differently. Twenty-eight Holstein cows were blocked for similar calving date 4wk before expected parturition to compare the effects of feeding sources of n-3 and n-6 fatty acids on milk production and composition, plasma metabolites, and liver parameters. Cows within each block were assigned to 1 of 3 isonitrogenous and isoenergetic diets: control with a source of calcium salts of palm oil (MEG; 1.1 and 2.6% of the dry matter in prepartum and postpartum diets, respectively); n-3 fatty acids supplied as whole flaxseed (WFL; 4.8 and 7.7% of the dry matter in prepartum and postpartum diets, respectively); and n-6 fatty acids supplied as whole linola (WLO; 4.8 and 7.7% of the dry matter in prepartum and postpartum diets, respectively). Diets were fed until wk 14 of lactation. Contrasts of WFL versus WLO and polyunsaturated fatty acids versus MEG were compared. Cows fed polyunsaturated fatty acids increased dry matter intake over time at a greater extent than those fed MEG, which resulted in enhanced energy balance. Cows fed MEG produced more milk compared with those fed polyunsaturated fatty acids, and there was no difference between those fed WFL and WLO. We found no effect on body condition score and body weight. Plasma concentrations of glucose, fatty acids, and BHB were similar among diets. There was no effect of diet on concentration of glycogen and activities of superoxide dismutase and glutathione peroxidase in the liver. We observed higher concentrations of hepatic lipids and triacylglycerol in cows fed MEG compared with those fed polyunsaturated fatty acids, and no difference between WFL and WLO. Hepatic catalase activity tended to be higher on wk 4 after calving for cows supplemented with WFL compared with those fed WLO. Feeding linoleic and linolenic acids as unprotected oilseeds increased dry matter intake over time at a greater extent for cows fed MEG, improved the energy status, and lowered hepatic lipids and triacylglycerol contents, which may contribute to enhance the health status of transition dairy cows.

Clinical response after chitosan microparticle administration and preliminary assessment of efficacy in preventing metritis in lactating dairy cows.

The objectives were to evaluate the clinical response to intrauterine administration of chitosan microparticles (CM) and to assess efficacy for preventing metritis in dairy cows. Holstein cows (n=104; 40 primiparous and 64 multiparous) at increased risk for metritis (cows that had abortion, dystocia, twins, stillbirth, or retained placenta) were randomly assigned to 1 of 2 treatments at 1d in milk (DIM; 24h postpartum): CM group (n=52), daily intrauterine infusion of 8g of CM dissolved in 40mL of sterile water for 5d; control (CON) group (n=52), daily intrauterine infusion of 40mL of sterile water for 5d. Clinical response was assessed by evaluation of parameters associated with inflammation (rectal temperature and plasma haptoglobin concentration) and metabolism [plasma nonesterified fatty acid (NEFA) and β-hydroxybutyrate (BHB) concentrations] up to 14 DIM, and daily milk yield up to 30 DIM. Uterine discharge pH was evaluated at 4, 7, 10, and 14 DIM as an indicator of bacterial load and acid byproduct production. The cumulative incidence of metritis was evaluated up to 4, 7, 10, and 14 DIM. Continuous and dichotomous outcomes were evaluated with mixed linear and logistic regression analysis, respectively. Treatment with CM did not affect rectal temperature (39.17±0.04 vs. 39.14±0.04°C), haptoglobin (1.10±0.05 vs. 1.07±0.05mg/mL), NEFA (0.64±0.04 vs. 0.63±0.04mmol/L), BHB (0.61±0.03 vs. 0.57±0.03mmol/L), or milk yield (30.3±0.92 vs. 30.1±0.97kg/d) compared with CON. An interaction between treatment and time showed that NEFA concentrations were lower for CM than CON at 10 DIM (0.46±0.06 vs. 0.64±0.06mmol/L). Treatment with CM resulted in greater uterine discharge pH than CON (6.91±0.03 vs. 6.83±0.02). Cows that developed metritis had increased concentrations of haptoglobin and BHB, and decreased uterine discharge pH and milk yield. Treatment with CM resulted in decreased incidence of metritis up to 7 DIM compared with CON (46.2 vs. 65.4%); however, no differences were found at 4 (11.5 vs. 17.3%), 10 (61.5 vs. 73.1%), and 14 DIM (63.5 vs. 73.1%) for CM versus CON, respectively. In conclusion, CM did not alter clinical parameters of cows at risk for metritis, and may merit further investigation for prevention of metritis. However, the duration of treatment may have to be extended to effectively reduce the incidence of metritis during the high-risk period.

Technical note: Comparison of 4 electronic handheld meters for diagnosing hyperketonemia in dairy cows

The objective of this study was to evaluate 4 handheld ketone meters for use in on-farm β-hydroxybutyrate (BHB) monitoring of hyperketonemia in transition dairy cows. Blood samples taken from 250 Holstein cows between 262d pregnant and 15d in milk were evaluated on 4 different handheld ketone meters: Precision Xtra (Abbott Laboratories, Abbott Park, IL), TaiDoc (Pharmadoc, Lüdersdorf, Germany), Nova Max (Nova Biomedical, Billerica, MA), and Nova Vet (Nova Biomedical). Samples were screened using the Precision Xtra and tested on the remaining 3 m if the sample BHB concentration fell into predetermined ranges. A total of 89 samples were used for analysis. Performance of each meter was compared with the average of 2 plasma BHB concentrations both determined by a gold standard spectrophotometric Randox assay performed at 2 independent laboratories. Agreement between the 2 laboratories was very strong (Pearson correlation=0.998). All meters had Pearson correlation coefficients greater than 0.95. The Precision Xtra and TaiDoc meters were 100.0% sensitive and 73.5% specific at a BHB concentration cut point of 1.2mmol/L. The Nova Vet and Nova Max meters had sensitivities of 94.9 and 74.4% and specificities of 91.8 and 100.0%, respectively, at the same cut point. Agreement between the gold standard and the handheld meter was the best for the Nova Vet meter when evaluated using a Bland Altman graph with a mean BHB difference of 0.08mmol/L. Trends in bias were noted with the Precision Xtra and Nova Max meters resulting in increasing average discrepancy between the gold standard and the meter for both at higher plasma BHB concentrations and mean BHB differences of -0.34 and 0.26mmol/L, respectively. The coefficient of variation was <10% for the Precision Xtra, TaiDoc, and Nova Vet meters, and <15% for the Nova Max meter. We conclude that the TaiDoc and Nova Vet meters, similar to the already validated Precision Xtra meter, are acceptable for use in on-farm testing for monitoring and treatment of hyperketonemia.

Effects of subcutaneous calcium administration at calving on mineral status, health, and production of Holstein cows

The objective of this study was to determine effects of subcutaneous (s.c.) infusions of Ca at calving day on serum concentrations of Ca, P, Mg, β-hydroxybutyrate (BHB), glucose, body condition score (BCS), milk yield, and health in fresh cows receiving a dietary cation-anion difference diet during the dry period. Three hundred seventy-five fresh Holstein cows were blocked based on parity (143 cows in first parity, 108 cows in second parity, and 124 cows in third or greater parity) and randomly assigned to 1 of 4 experimental treatments immediately after calving. Treatments were control group without infusion (control; n=190); 1 s.c. infusion of 250mL of 40% Ca borogluconate (1SC250; n=72) at calving; 1 s.c. infusion of 500mL of 40% Ca borogluconate (1SC500; n=63) at calving; 2 s.c. infusions of 250mL of 40% Ca borogluconate, one immediately after calving and the second 12 to 18h after first infusion (2SC250; n=50). Blood samples were collected immediately after parturition and at 1, 2, 4, and 7d in milk (DIM) for Ca, P, and Mg determination. Milk production, milk composition, and somatic cell count were recorded monthly up to 90 DIM. The evaluation of BCS was performed at calving and at 38 DIM. A subset of 9 cows per treatment group was randomly chosen to measure serum concentration of glucose and BHB at 2, 4, and 7 DIM. Total serum Ca in 1SC250 (8.95mg/dL), 1SC500 (9.27mg/dL), and 2SC250 (9.07mg/dL) was greater during the first week postpartum compared with control (8.45mg/dL). Serum concentrations of P, Mg, BHB, glucose, and milk yield were not affected by treatments. The dry matter intake during the first 24h after calving was higher for treatments 1SC250 (13.5kg), 1SC500 (15.0kg), and 2SC250 (15.6kg) relative to control (12.5kg). Milk somatic cell counts were lower for 1SC500 (90.5 cells/mL) and 2SC250 (82.2 cells/mL) than control (132.8 cells/mL). Risk ratio was >1 for development of metritis, and clinical and subclinical endometritis in control cows relative to 2SC250 cows, which indicates a higher risk of developing disease for control cows (2.03, 1.7, and 1.8 times, respectively). These results suggest that prophylactic s.c. infusions of Ca at calving can improve postpartum Ca status in Holstein cows and intake at 1 DIM. Given the reduction of metritis, clinical and subclinical endometritis, and subclinical hypocalcemia with treatment, the effect of s.c. Ca supplementation on immune status warrants further investigation.

Prevalence of elevated milk β-hydroxybutyrate concentrations in Holstein cows measured by Fourier-transform infrared analysis in Dairy Herd Improvement milk samples and association with milk yield and components

The purpose was to describe the prevalence and effect of elevated milk β-hydroxybutyrate (BHB) as detected by routine Fourier-transform infrared analysis in Dairy Herd Improvement milk samples. Data collected over 4 yr included cow information as well as milk yield and composition from 498,310 samples from postparturient Holstein cows (5-35d in milk) from 4,242 herds. The following thresholds were used to classify cows based on their early lactation milk BHB concentration: <0.15mmol/L=negative; 0.15 to 0.19mmol/L=suspect; and ≥0.20mmol/L=positive. Overall prevalence (suspect + positive) was 22.6% and was higher for older cows (18.7, 19.5, and 27.6%, for cows in their first, second, and third or greater lactation, respectively). Distribution with regards to days in milk was different among parity groups, with first-lactation cows having highest prevalence (30%) in the first week after calving; cows in their second and third and greater parity had the highest prevalence in the second week after calving, at 25.8 and 34.6%, respectively. Season of calving affected the prevalence of elevated milk BHB, with cows calving in the fall and spring seasons showing higher prevalence. Distribution among herds was highly variable, as 45% of herds had a prevalence of 20% or less, 47% of herds had a prevalence between 21 and 40%, 6% of herds had a prevalence between 40 and 50%, and 2% of herds had a prevalence of 50% or above. Positive cows had lower milk yield, protein concentration and yield, and lower Transition Cow Index than negative cows, but also higher fat concentration and yield, as well as higher somatic cell count than negative cows. Suspect cows were generally intermediate. The present analysis highlights the opportunity for elevated milk BHB monitoring at the herd level through routine BHB testing in Dairy Herd Improvement milk samples.

Short communication: Associations between blood fatty acids, β-hydroxybutyrate, and α-tocopherol in the periparturient period in dairy cows: An observational study

The objective of the present study was to examine the relationships between blood concentrations of fatty acids, β-hydroxybutyrate (BHB), and α-tocopherol during the periparturient period in dairy cows. Blood samples were collected from 131 cows belonging to 4 different commercial dairy farms in southeastern Europe (Greece and Italy). We determined blood concentrations of fatty acids, BHB, and α-tocopherol at dry-off, at calving, and 30d postpartum. Results indicated that fatty acid concentrations were low at dry-off, reached maximum value at calving, and then declined at 30d postpartum. In fact, fatty acid concentrations at 30d postpartum were 50% lower than at calving. In contrast, BHB concentrations were low at dry-off, increased by 27% at calving, and continued to increase by another 20% at 30d postpartum. Overall, we found a weak correlation between fatty acids and BHB throughout the periparturient period. Concentrations of α-tocopherol were lowest at calving, and we detected no differences in α-tocopherol concentrations at dry-off or 30d postpartum. Negative correlations between fatty acids and α-tocopherol were highly significant at 30d postpartum and approached the level of significance at dry-off. However, both correlations became nonsignificant following the adjustment of α-tocopherol with cholesterol, indicating that the correlations were a reflection of changes in lipid transport. We found significant negative correlations (strong at dry-off and weak at 30d postpartum) between BHB and α-tocopherol after adjustment with cholesterol. The physiological basis for the negative correlations between BHB and α-tocopherol, especially that at dry-off, is not known and should not be taken to imply a cause-effect relationship. However, it opens the door to investigating the effects of vitamin E on liver function in dairy cows.

Measurement of β-hydroxybutyrate in capillary blood obtained from an ear to detect hyperketonemia in dairy cows by using an electronic handheld device

The primary objective of the present study was to test whether capillary blood obtained by puncturing the skin of an ear with a minimal invasive lancet technique is able to detect hyperketonemia (HYK) in dairy cows. Furthermore, test characteristics of a new available handheld device, the FreeStyle Precision Neo (FSP-Neo, Abbott GmbH & Co. KG, Wiesbaden, Germany) for determination of β-hydroxybutyrate (BHB) concentrations in bovine blood were evaluated by comparing the measurements with a laboratory reference. The BHB concentration was determined with the FSP-Neo device in 720 capillary blood samples from 3 different sampling sites (left, right ear, and repeated measurement) and in 240 samples from a coccygeal vessel. The concentration of BHB in serum harvested from the coccygeal blood samples was analyzed at the laboratory and was used as reference. The Spearman correlation coefficient (ρs) between the BHB concentrations in capillary blood measured with the handheld device and the reference test was between 0.76 and 0.81. Using capillary blood, the mean ± standard deviation BHB difference compared with the reference test was 0.20±0.47 mmol/L for all 3 sampling locations at the ears. The receiver operating characteristic analyses for the FSP-Neo device resulted in an optimized threshold for the detection of subclinical ketosis (SCK) in capillary blood of 1.3 mmol/L (left and right ear) and 1.2 mmol/L (repeated measurements). Applying these adjusted threshold sensitivities (Se) for all 3 capillary sampling sites at the ear were 100%, and specificities (Sp) ranged between 93 and 94%. Hence, we conclude that all sampling locations were suitable to identify cows suffering from SCK. The reference test compared with BHB measurements in coccygeal blood resulted in a ρs of 0.92 with a mean ± standard deviation of 0.02±0.21 mmol/L. The receiver operating characteristic analyses for the FSP-Neo device resulted in an optimized threshold for the detection of SCK in coccygeal blood of 1.1 mmol/L, with a corresponding Se and Sp of 100 and 95%, respectively. Because capillary blood is easily achievable from an ear, particularly if animals are fixed in headlocks for routine checkups, this technique is considered as an additional minimally invasive method for the identification of dairy cows suffering from HYK.

Estimation of the cow- and herd-level prevalence of post-partum subclinical ketosis in large Washington state dairy herds and evaluation of mean β-hydroxybutyrate concentration for herd-level assessment

The purpose of this study was to estimate the cow- and herd-level prevalence of subclinical ketosis (SCK) on dairies in the state of Washington and evaluate the arithmetic mean of individual samples as a proxy for pooled-sample testing to screen herds for SCK. The cow-level prevalence of SCK in 589 cows from 40 Washington dairies was 31,25, and 19% using P-hydroxybutyrate (BHB) concentration cutpoints of &gt; 1.0, 1.2, and 1.4 mmol/L, respectively. For all BHB cutpoints, no difference was observed in the prevalence of SCK between cows of parity 1 and 2 (P &gt; 0.358), but SCK was higher in cows of parity &gt; 3 versus those in lactations 1 and 2 (P &lt;0.001). The mean BHB concentration (mmol/L) in cows of parity &gt; 3 (1.16) was higher than that of parity 1 (0.72) and parity 2 (0.88) cows (P &lt;0.001). Using a BHB cutpoint of &gt; 1.2 mmol/L and herd alarm level of &gt;15% cows exceeding that cutpoint, 23 herds (58%) demonstrated a herd-level problem with SCK. When a BHB cutpoint of &gt; 1.4 and herd alarm level of &gt;25% was used, 12 herds (30%) had SCK. A herd mean BHB concentration of 0.77 mmol/L was correlated with a 15% herd-alarm level using a &gt; 1.2 mmol/L BHB cutpoint. Sensitivity and specificity of the herd mean BHB cutpoint of &gt;0.8 mmol/L to identify herds with &gt; 15% cows with BHB &gt; 1.2 mmol/L was 91 and 75%, respectively. The prevalence of SCK in Washington dairies was numerically higher than previous reports. Results of this study highlight the importance of obtaining representative samples from parity risk groups. Furthermore, results suggest parity 2 cows may be better grouped with parity 1 rather than &gt; 3 parity cows. Herd mean BHB concentration performed well as a test to identify herds with a potential SCK problem when a mean value specific cutpoint was used. Further research evaluating the relationship between pooled-sample BHB and important outcomes, such as disease, milk production, reproduction, and removal, are needed.

Insulin signaling, inflammation, and lipolysis in subcutaneous adipose tissue of transition dairy cows either overfed energy during the prepartum period or fed a controlled-energy diet

Adipose tissue mobilization is a hallmark of the transition period in dairy cows. Cows overfed energy during the dry period have higher concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) compared with cows fed a controlled-energy diet prepartum. The reason for an increase in blood NEFA concentrations at the level of adipose tissue in cows overfed energy has not been fully elucidated. One hypothesis is that cows with high BHB concentrations suffer from adipose tissue-specific insulin resistance, leading to higher rates of adipose tissue mobilization in the postpartum period. To test this hypothesis, subcutaneous adipose tissue biopsies of cows overfed energy in excess of predicted requirements by 50% in the dry period, and that had high concentrations of blood BHB postpartum (group H; n=12), were used. Findings were compared with results of biopsies from cows fed a controlled-energy diet and with low BHB concentrations postpartum (group C; n=12) to create the biggest contrast in BHB concentrations. Subcutaneous adipose tissue biopsies were obtained before and 60 min after an intravenous glucose challenge (0.25 g/kg of glucose) at 28 and 10 d before expected calving as well as on d 4 and 21 postpartum. Phosphorylation of protein kinase B, extracellular signal-regulated kinase, and hormone-sensitive lipase was determined before and after glucose infusion by Western blot. Western blot was also used to assess the baseline protein abundance of peroxisome proliferator-activated receptor gamma and insulin receptor β-subunit. In addition, gene expression of fatty acid synthase, adiponectin, monocyte chemoattractant protein 1, and tumor necrosis factor α was determined by real-time quantitative reverse-transcription PCR. Backfat thickness was determined in the thurl area by ultrasonography. Cows in group H showed a greater degree of lipogenesis prepartum, but no differences were found in lipolytic enzyme activity postpartum compared with cows in group C. Baseline plasma insulin concentrations were decreased and serum NEFA concentrations increased postpartum in group H. Insulin signaling through protein kinase B, quantity of insulin receptor, markers of inflammation, and peroxisome proliferator-activated receptor gamma in adipose tissue were not different between the groups, but expression of adiponectin was increased in adipose tissue of cows in group H during the immediate peripartum period. In conclusion, differences in serum concentrations of NEFA between cows overfed energy prepartum and high blood concentrations of BHB are likely due to greater negative energy balance postpartum reflected in lower circulating concentrations of glucose and insulin and an increase in the total amount of mobilized adipose tissue mass rather than due to changes in adipose tissue insulin signaling.

Short communication: Association of milk fatty acids with early lactation hyperketonemia and elevated concentration of nonesterified fatty acids

The objective of our study was to extend the limited research available on the association between concentrations of milk fatty acids and elevated nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) concentrations in early lactation dairy cattle. Measurement of milk fatty acids for detection of cows in excessive negative energy balance has the potential to be incorporated in routine in-line monitoring systems. Blood samples were taken from 84 cows in second or greater lactation 3 times per week between 3 to 14 d in milk. Cows were characterized as hyperketonemic (HYK) if blood BHB concentration was ≥1.2mmol/L at least once and characterized as having elevated concentrations of NEFA (NEFAH) if serum NEFA concentration was ≥1mmol/L at least once. Composition of colostrum and milk fatty acids at wk 2 postpartum was used to investigate the potential diagnostic value of individual fatty acids and fatty acid ratios for the correct classification of cows with NEFA and BHB concentrations above these thresholds, respectively. Receiver operating characteristic (ROC) curves were used to identify thresholds of fatty acid concentration and fatty acid ratios when ROC area under the curve was ≥0.70. Correct classification rate (CCR, %) was calculated as {[(number of true positives + number of true negatives)/total number tested] × 100}. None of the colostrum fatty acids yielded a sufficiently high area under the curve in ROC analysis for the association with HYK and NEFAH. The following fatty acids and fatty acid ratios were identified for an association with NEFAH (threshold, CCR): C15:0 (≤0.65g/100g, 68.3%); cis-9 C16:1 (≥1.85g/100g, 70.7%); cis-9 C18:1 (≥26g/100g, 69.5%), cis-9 C18:1 to C15:0 ratio (≥45, 69.5%); cis-9 C16:1 to C15:0 (≥2.50, 73.2%). Several fatty acids were associated with HYK (threshold, CCR): C6:0 (≤1.68g/100g, 80.5%), C8:0 (≤0.80g/100g, 80.5%), C10:0 (≤1.6g/100g, 79.3%); C12:0 (≤1.42g/100g, 82.9%); C14:0 (≤6.10g/100g, 84.1%); C15:0 (≤0.50g/100g, 82.9%), cis-9 C18:1 (≥30g/100g, 81.7%). The use of fatty acid ratios did not improve CCR over using individual fatty acids for the classification of HYK. Colostrum fatty acid composition was not useful in predicting NEFAH or HYK between 3 to 14 d in milk. Accuracy of milk fatty acids and fatty acid ratios to correctly classify cows with elevated concentrations of NEFA and BHB between 3 to 14 d in milk was moderate and overall higher for HYK. Determining changes in the fatty acid composition of milk fat from milk samples at wk 2 postpartum for the detection of cows with elevated concentrations of BHB and NEFA can currently not be recommended to replace direct measurement. Future applications should target repeated milk sampling between 3 to 14 d in milk to identify the best sampling for determination of milk fatty acid composition within the first 2 wk postpartum.

Bicarbonate correction of ketoacidosis alters host-pathogen interactions and alleviates mucormycosis.

Patients with diabetic ketoacidosis (DKA) are uniquely predisposed to mucormycosis, an angioinvasive fungal infection with high mortality. Previously, we demonstrated that Rhizopus invades the endothelium via binding of fungal CotH proteins to the host receptor GRP78. Here, we report that surface expression of GRP78 is increased in endothelial cells exposed to physiological concentrations of β-hydroxy butyrate (BHB), glucose, and iron that are similar to those found in DKA patients. Additionally, expression of R. oryzae CotH was increased within hours of incubation with DKA-associated concentrations of BHB, glucose, and iron, augmenting the ability of R. oryzae to invade and subsequently damage endothelial cells in vitro. BHB exposure also increased fungal growth and attenuated R. oryzae neutrophil-mediated damage. Further, mice given BHB developed clinical acidosis and became extremely susceptible to mucormycosis, but not aspergillosis, while sodium bicarbonate reversed this susceptibility. BHB-related acidosis exerted a direct effect on both GRP78 and CotH expression, an effect not seen with lactic acidosis. However, BHB also indirectly compromised the ability of transferrin to chelate iron, as iron chelation combined with sodium bicarbonate completely protected endothelial cells from Rhizopus-mediated invasion and damage. Our results dissect the pathogenesis of mucormycosis during ketoacidosis and reinforce the importance of careful metabolic control of the acidosis to prevent and manage this infection.

Effects of dry period length and dietary energy source on milk yield, energy balance, and metabolic status of dairy cows over 2 consecutive years: Effects in the second year

The objective of the current study was to evaluate the effect of dry period (DP) length on milk yield, energy balance (EB), and metabolic status in cows fed a lipogenic or glucogenic diet in the second year after implementation of DP and dietary treatments. Holstein-Friesian dairy cows (n=167) were assigned randomly to 1 of 3 DP lengths (0, 30, or 60d) and 1 of 2 early lactation diets (glucogenic or lipogenic) for 2 consecutive years. Results of the first year were reported previously. In the second year, 19 cows in the 0-d DP group were attributed to a new group (0→67d DP) because these cows had a milk yield of <4kg/d at least 30d before expected calving date and were dried off. Milk yield was recorded and EB was calculated from wk –8 to 9 relative to calving. Blood samples were taken weekly from wk –3 to 8 relative to calving. Liver samples were taken in wk –2, 2, and 4 relative to calving. At the onset of lactation, cows with a 0-d or 0→67-d DP had greater body condition score (BCS) than cows with a 60-d DP. During the first 9wk, cows with a 0- or 30-d DP produced 5.0 and 4.3kg less milk per day, respectively, but had similar EB compared with cows with a 60-d DP. Cows with a 0- or 30-d DP produced additional milk precalving, which could compensate milk yield losses in the first 9wk postcalving. Cows with a 0-d DP did not have milk yield losses or improve EB in the second year as much as in the first year. Cows with a 0-d DP had greater plasma insulin and insulin-like growth factor-I (IGF-I) and lower liver triacylglycerol concentrations than cows with other DP lengths. Cows with a 0→67-d DP had lower EB, and greater plasma free fatty acids (FFA) and β-hydroxybutyrate (BHB) concentrations than cows with other DP lengths. Feeding a glucogenic diet increased plasma glucose, IGF-I, and insulin concentrations, and decreased plasma FFA, BHB, and urea concentrations compared with a lipogenic diet, independent of DP length. In conclusion, omitting the DP or feeding a glucogenic diet improved metabolic status in early lactation of the second year after implementation of DP length and dietary treatments, although effects of omitting the DP were less pronounced in the second year than in the first year. The less pronounced improvement of EB in the second year was related to the high BCS at the onset of lactation and reduced milk yield losses in cows with a 0-d DP.

Prepartum supplementation of nicotinic acid: Effects on health of the dam, colostrum quality, and acquisition of immunity in the calf

Nicotinic acid (NA) has been shown to reduce lipolysis, alter milk components and the ruminal environment, and increase blood flow. Increased blood flow to the mammary gland during colostrogenesis might increase nutrients and immunoglobulin concentration of colostrum. Twenty-six multiparous Holstein cows were housed in a tiestall barn. Cows were blocked by expected calving date and randomly assigned to 1 of 2 treatments 4 wk prepartum: (1) 0g/d of NA (control, CON) or (2) 48g/d of NA (NA). Total mixed ration amounts fed and refused were measured daily to determine dry matter intake. Blood samples were collected from dams every Monday, Wednesday, and Friday from the coccygeal vein or artery and were analyzed for glucose, nonesterified fatty acids (NEFA), and β-hydroxybutyrate (BHB). Colostrum was collected and weighed within 90 min of parturition. Colostral immunoglobulin G (IgG) concentration was analyzed using radial immunodiffusion assay. Calves were removed from their dams before suckling and weighed within 30 min after birth. Calves received 3 L of a lacteal-based colostrum replacer that provided a total of 225.8g of IgG within 2h of birth. Calf blood samples were collected via jugular venipuncture at 0 and 24h of age and analyzed for IgG concentration and determination of apparent efficiency of absorption. Colostrum yield, dry matter intake, IgG yield, and fat and solids percentage of colostrum did not differ between treatments. Serum concentrations of glucose and BHB were not affected by treatment. We detected an effect of week on serum glucose concentrations at calving and on serum BHB concentrations at 1 wk postpartum. There was a treatment by week effect for serum NEFA concentrations at 1 wk postpartum, where cows that received NA prepartum had higher serum NEFA concentration than CON cows, indicating that a NEFA rebound occurred. No differences were observed for calf body weight, 0- or 24-h serum IgG concentration, or apparent efficiency of absorption. Supplementation of NA increased IgG concentration in colostrum from 73.8 to 86.8g/L. Results indicate that 48g/d of supplemental NA during the prepartum period improved colostrum quality.

The Population Pharmacokinetics of D-β-hydroxybutyrate Following Administration of (R)-3-Hydroxybutyl (R)-3-Hydroxybutyrate.

The administration of ketones to induce a mild ketosis is of interest for the alleviation of symptoms associated with various neurological disorders. This study aimed to understand the pharmacokinetics (PK) of D-β-hydroxybutyrate (BHB) and quantify the sources of variability following a dose of (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (ketone monoester). Healthy volunteers (n = 37) were given a single drink of the ketone monoester, following which, 833 blood BHB concentrations were measured. Two formulations and five dose levels of ketone monoester were used. A nonlinear mixed effect modelling approach was used to develop a population PK model. A one compartment disposition model with negative feedback effect on endogenous BHB production provided the best description of the data. Absorption was best described by two consecutive first-order inputs and elimination by dual processes involving first-order (CL = 10.9L/h) and capacity limited elimination (V max = 4520mg/h). Covariates identified were formulation (on relative oral bioavailable fraction and absorption rate constant) and dose (on relative oral bioavailable fraction). Lean body weight (on first-order clearance) and sex (on apparent volume of distribution) were also significant covariates. The PK of BHB is complicated by complex absorption process, endogenous production and nonlinear elimination. Formulation and dose appear to strongly influence the kinetic profile following ketone monoester administration. Further work is needed to quantify mechanisms of absorption and elimination of ketones for therapeutic use in the form of ketone monoester.

Quelling Inflammation with Ketosis and Steric Chemistry.

Quelling Inflammation with Ketosis and Steric Chemistry.

Dry period plane of energy: Effects on glucose tolerance in transition dairy cows

Overfeeding energy in the dry period can affect glucose metabolism and the energy balance of transition dairy cows with potential detrimental effects on the ability to successfully adapt to early lactation. The objectives of this study were to investigate the effect of different dry cow feeding strategies on glucose tolerance and on resting concentrations of blood glucose, glucagon, insulin, nonesterified fatty acids (NEFA), and β-hydroxybutyrate (BHB) in the peripartum period. Cows entering second or greater lactation were enrolled at dry-off (57d before expected parturition) into 1 of 3 treatment groups following a randomized block design: cows that received a total mixed ration (TMR) formulated to meet but not exceed energy requirements during the dry period (n=28, controlled energy); cows that received a TMR supplying approximately 150% of energy requirements during the dry period (n=28, high energy); and cows that were fed the same diet as the controlled energy group for the first 28d, after which the TMR was formulated to supply approximately 125% of energy requirements until calving (n=28, intermediate energy). Intravenous glucose tolerance tests (IVGTT) with rapid administration of 0.25g of glucose/kg of body weight were performed 28 and 10d before expected parturition, as well as at 4 and 21d after calving. Area under the curve for insulin and glucose, maximal concentration and time to half-maximal concentration of insulin and glucose, and clearance rates were calculated. Insulin resistance (IR) indices were calculated from baseline samples obtained during IVGTT and Spearman rank correlations determined between IVGTT parameters and IR indices. Treatment did not affect IVGTT parameters at any of the 4 time points. Correlation between IR indices and IVGTT parameters was generally poor. Overfeeding cows energy in excess of predicted requirements by approximately 50% during the entire dry period resulted in decreased postpartum basal plasma glucose and insulin, as well as increased glucagon, BHB, and NEFA concentrations after calving compared with cows fed a controlled energy diet during the dry period. In conclusion, overfeeding energy during the entire dry period or close-up period alone did not affect glucose tolerance as assessed by IVGTT but energy uptake during the dry period was associated with changes in peripartal resting concentrations of glucose, as well as postpartum insulin, glucagon, NEFA, and BHB concentrations.

Response to dietary-induced energy restriction in dairy sheep divergently selected for resistance or susceptibility to mastitis

Dairy ruminants experiencing a severe postpartum negative energy balance (NEB) are considered to be more susceptible to mastitis. Although the genetic variability of mastitis resistance is well established, the biological basis of the link between energy metabolism and resistance is mostly unknown. The aim of this study was to characterize the effect of NEB on metabolism and immune response according to the genetic background for mastitis resistance or susceptibility. Forty-eight ewes from high and low somatic cell score (SCS) genetic lines were allocated to 2 homogeneous subgroups 2 wk after lambing: one group (NEB) received an energy-restricted diet to cover 60% of their energy requirements, and the other group received a control (positive energy balance: PEB) diet. Both diets met the protein requirements. After 10 d on either the NEB or PEB diet, all ewes were injected with a Pam3CSK4/MDP solution in one half-udder to induce an inflammatory response. The ewes were monitored for milk production, somatic cell count (SCC), body weight (BW), body condition score (BCS), and blood metabolites. Differential milk cell counts were determined by flow cytometry. Plasma concentrations of glucose, insulin, nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHB), and triiodothyronine were determined. Energy restriction resulted in an increased fat:protein ratio in milk and decreased milk yield, BW, and BCS. The NEB ewes had significantly higher NEFA and BHB and lower plasma glucose concentrations than PEB ewes, reflecting a mobilization of body reserves and ketone body synthesis. High-SCS ewes had a higher SCS than low-SCS throughout the experiment, except after the inflammatory challenge, which resulted in similar SCS in all 4 groups. A noteworthy interaction between genetic background and diet was evidenced on metabolic parameters and BW. Indeed, high-SCS ewes subjected to NEB showed greater decrease in BW and increased NEFA and BHB concentrations compared with low-SCS ewes. Thus, NEB in early lactation led to extensive mobilization of body reserves and intense ketone body synthesis in mastitis-susceptible sheep. These results reinforce the hypothesis of a genetic association between mastitis susceptibility and energy metabolism and open the way to further studies on the biological basis for this association.

Hot topic: Early postpartum treatment of commercial dairy cows with nonsteroidal antiinflammatory drugs increases whole-lactation milk yield

Previous research has shown that postpartum administration of the nonsteroidal antiinflammatory drug (NSAID) sodium salicylate can increase 305-d milk yield in older dairy cattle (parity 3 and greater). However, in this prior work, sodium salicylate was delivered to cows via the drinking water, a method that does not align well with current grouping strategies on commercial dairy farms. The objective of the current study was to replicate these results on a commercial dairy farm with a simplified treatment protocol and to compare sodium salicylate with another NSAID, meloxicam. Dairy cattle in their second lactation and greater (n=51/treatment) were alternately assigned to 1 of 3 treatments at parturition, with treatments lasting for 3d. Experimental treatments began 12 to 36h after parturition and were (1) 1 placebo bolus on the first day and 3 consecutive daily drenches of sodium salicylate (125g/cow per day; SAL); (2) 1 bolus of meloxicam (675mg/cow) and 3 drenches of an equal volume of water (MEL); or (3) 1 placebo bolus and 3 drenches of water (CON). Blood samples were collected on the first day of treatment, immediately following the last day of treatment, and 7d after the last day of treatment; plasma was analyzed for glucose, β-hydroxybutyrate (BHB), free fatty acids, haptoglobin, and paraoxonase. Milk production, body condition score, reproductive status, and retention in the herd were monitored for 365d posttreatment, and effects of treatment, parity, days in milk, and interactions were evaluated in mixed effects models. Significance was declared at P<0.05. Whole-lactation milk and protein yields were greater in NSAID-treated cows, although 305-d fat production was not affected. There was a significant interaction of treatment and parity for plasma glucose concentration; MEL increased plasma glucose concentrations compared with CON and SAL in older cows. Sodium salicylate decreased plasma BHB concentration compared with MEL at 7d posttreatment, although no difference was detected immediately following treatment. Haptoglobin concentrations were elevated in SAL cows compared with CON. There was a tendency for CON cows to be removed from the herd more quickly than MEL cows (42 vs. 26% at 365d posttreatment). Body condition score, concentrations of plasma free fatty acids and paraoxonase, and time to pregnancy were not affected by treatment. These results indicate that NSAID administration in postpartum cows has the potential to be a viable way to improve productivity and potentially longevity in commercial dairies, although further research is necessary to optimize recommendations for producers.