Blood Blood Β-hydroxybutyrate Research Articles

Postpartum health is associated with detection of estrus by activity monitors and reproductive performance in dairy cows

The objective of this prospective observational study was to investigate associations of postpartum health with estrus detection (ED) by activity monitors and pregnancy outcomes in dairy cows. A total of 1,743 Holstein cows from 2 commercial dairy herds in Ontario, Canada were enrolled 3 wk before expected parturition and examined for health variables until 9 wk postpartum. Body condition score (BCS) and lameness were measured at 3 wk prepartum, and serum concentrations of total Ca, haptoglobin (Hp), and nonesterified fatty acids were measured at 2 and 6 ± 2 d in milk (DIM), and blood β-hydroxybutyrate (BHB) and metritis were assessed at 4, 8, 11, and 15 ± 2 DIM. Cows were examined for purulent vaginal discharge (PVD) and endometritis (ENDO) by endometrial cytology at wk 5, for lameness at wk 3 and 7, for BCS at wk 9 postpartum, and for time to onset of cyclicity by biweekly serum progesterone (P4) measurements. Additional disease data were obtained from farm records. Reproductive management for first AI was primarily based on ED by activity monitors until at least 75 DIM, and cows not detected in estrus were synchronized. Data were analyzed in multivariable logistic or Cox proportional hazards regression models including blood markers, health variables, potential covariates, and herd as a random effect. Estrus was detected in 77% of primiparous and 66% of multiparous cows between 50 or 55 DIM and 75 DIM. In 1,246 cows, the model-predicted probability of ED (percentage point difference) was lower in cows that had retained placenta (-14%), ENDO (-7%), PVD (-8%), delayed cyclicity (no P4 > 1 ng/mL by wk 9; -12%), or ≥0.5-point BCS loss (-14%) compared with cows without each of these risk factors, and it was negatively associated with blood BHB at 15 DIM. Considering only variables measured on farm (not requiring laboratory analysis), the probability of ED was lower (56 vs. 81%) in cows with >1 risk factor compared with cows without risk factors. The predicted probability of pregnancy at first artificial insemination (percentage point difference) was lower in cows that had ENDO (-7%) or PVD (-7%), and negatively associated with serum Hp at 6 ± 2 DIM. In cows detected in estrus by 75 DIM (n = 888), risk factors for reduced pregnancy rate by 250 DIM (adjusted hazard ratio (AHR); 95% confidence intervals) included difficult calving (AHR: 0.67; 0.45 to 1.00), metritis (AHR: 0.79; 0.61 to 1.01), PVD (AHR: 0.79; 0.65 to 0.97), or lameness (AHR: 0.79; 0.62 to 1.01), and it was negatively associated with serum Hp at 6 ± 2 DIM. Monitoring postpartum health may be used to identify cows that are more or less likely to be detected in estrus by activity monitors and to become pregnant in a timely manner. This would support a selective reproductive management program with targeted interventions.

Breath Analysis for Early Detection of Rising Ketone Bodies in Postpartum Dairy Cows Classified as at Risk of Ketosis

Ketosis is a metabolic disorder associated with high production and low energy intake in dairy cows. Fat reserves are mobilized, and gluconeogenesis occurs. Traditionally, rapid tests for blood, milk or urine are used to detect increased ketone body levels in case of ketosis. Breath analysis is relatively new and relevant for the development of automatic sensor systems for early warning. This study aims to determine whether and when a postpartum rise in ketone bodies occurs in breath with elevated blood β-hydroxybutyrate (BHB) in cows at risk of ketosis. Postpartum breath, blood, urine and milk samples were taken daily until day 7, with one prepartum sample excluding milk, and ketone body concentrations were determined. Concentrations of blood BHB were 0.4–2.6 mmol/L (lab) and 0.3–3.1 mmol/L (rapid test), breath acetone was 2.3–20.0 ppm, urine acetoacetate 0–8 mmol/L and milk BHB 0–500 µmol/L. A rise in blood BHB was related to that in urine acetoacetate and milk BHB and followed by a rise in breath acetone. However, breath acetone levels of ketotic and non-ketotic cows were similar. We conclude that rising ketone bodies can be detected in blood, urine, milk and breath, but to use breath analysis as an alternative to rapid tests, longitudinal sampling is needed.

Relação gordura: proteína do leite no primeiro controle leiteiro pós-parto em vacas leiteiras

ABSTRACT: This study evaluated milk fat to protein ratio (FPR) in the first test-day after calving of dairy cows in Paraná State. Data from 257,847 first monthly test-days after calving of 114,162 cows were submitted to analysis after the data edition. Procedures MIXED, CORR and STEPWISE from SAS were used in the data analysis. In order to validate FPR, a herd with regular postpartum monitoring of blood β-hydroxybutyrate (BHB) was used to estimate the correlation with FPR. There was a strong positive correlation between FPR and fat content (r = 0.85; P < 0.01) and a weak negative correlation between FPR and protein content (r = - 0.23; P < 0.01). The regression equation that best fitted FPR was 1.1806 + 0.3304*%F - 0.3877*%P (R2 = 0.98), where the variable with the greatest influence was milk fat content (partial R2 = 0.72). Animals of 4th and 5th or more lactations had higher (P < 0.01) FPR, followed by animals of third, first, and second lactations. Jersey cows had higher (P < 0.01) FPR than Holsteins and Brown Swiss animals. Cows milked twice daily had higher (P < 0.01) FPR than animals milked three times daily. There were small positive correlations between milk FPR and blood BHB on days 4, 7 and 12 after calving (0.07, 0.13, and 0.14, respectively). In conclusion, milk fat content was reported to be the most important variable affecting FPR changes, but the milk FPR has limited value to evaluate hyperketonemia incidence during the transition period.

The effects of dexamethasone administration on physiological, behavioral, and production parameters in dairy cows after a difficult calving.

Dairy cows are predisposed to diseases during the postpartum period. Dystocia has been associated with increased risk for disease, which is likely the result of increased tissue trauma and stress during the prolonged parturition. To attenuate the inflammatory response seen in dystocic animals and improve well-being, we assessed the effects of a glucocorticoid, dexamethasone administered within 12 h after calving. Dystocia was defined as a difficult birth resulting in a prolonged calving (≥70 min after the amniotic sac appears) and was monitored through 3 video cameras in the close-up dry-cow pen. Cows meeting the dystocia definition were randomly assigned to receive a single intramuscular injection of either dexamethasone (DEX; 0.1 mg/kg of body weight; n = 43) or saline (CON, n = 44) within 12 h following a dystocic calving. Serum haptoglobin, blood β-hydroxybutyrate (BHB) concentrations, body temperature, and several behaviors were measured for the first 7 d postpartum. Additionally, milk production and components for the first 120 d were recorded. Using a mixed model, the fixed effects of treatment, parity, calving assistance, and time, along with 2- and 3-way interactions, were analyzed with cow as a random effect. We observed that primiparous DEX cows had greater serum haptoglobin concentrations on d 3 and d 7 postpartum compared with primiparous CON cows. There was no difference between treatment groups for blood BHB concentrations and body temperature. Behavior was altered between treatments, with DEX cows having reduced activity for the first week postpartum, as well as less restlessness and increased lying times on some of the days following calving. Treatment interacted with time for milk yield, such that DEX cows produced 2.7 kg/d less milk than CON cows for the first month following calving. The administration of dexamethasone resulted in changes in behavioral measurements, which could suggest a reduction in discomfort; however, due to the reduction in milk yield for the first month following calving, DEX administration may not be applicable for typical farm use. Additional research is needed to investigate treatments for cows experiencing dystocia without detrimental effects on milk yield.

The association of hyperketonemia with fecal and rumen microbiota at time of diagnosis in a case-control cohort of early lactation cows

BackgroundMany dairy cows experience a state of energy deficit as they transition from late gestation to early lactation. The aims of this study were to 1) determine if the development of hyperketonemia in early lactation dairy cows is indicated by their gut microbiome, and 2) to identify microbial features which may inform health status. We conducted a prospective nested case-control study in which cows were enrolled 14 to 7 days before calving and followed through their first 14 days in milk (DIM). Hyperketonemic cows (HYK, n = 10) were classified based on a blood β-hydroxybutyrate (BHB) concentration 1.2 mmol/L within their first 14 DIM. For each HYK cow, two non-HYK (CON, n = 20) cows were matched by parity and 3 DIM, with BHB < 1.2 mmol/L. Daily blood BHB measures were used to confirm CON cows maintained their healthy status; some CON cows displayed BHB 1.2 mmol/L after matching and these cows were reclassified as control-HYK (C-HYK, n = 9). Rumen and fecal samples were collected on the day of diagnosis or matching and subjected to 16S rRNA profiling.ResultsNo differences in taxa abundance, or alpha and beta diversity, were observed among CON, C-HYK, and HYK health groups for fecal microbiomes. Similar microbiome composition based on beta diversity analysis was detected for all health statuses, however the rumen microbiome of CON and HYK cows were found to be significantly different. Interestingly, highly similar microbiome composition was observed among C-HYK cow rumen and fecal microbiomes, suggesting that these individual animals which initially appear healthy with late onset of hyperketonemia were highly similar to each other. These C-HYK cows had significantly lower abundance of Ruminococcus 2 in their rumen microbiome compared to CON and HYK groups. Multinomial regressions used to compute log-fold changes in microbial abundance relative to health status were not found to have predictive value, therefore were not useful to identify the role of certain microbial features in predicting health status.ConclusionsLower relative abundance of Ruminococcus 2 in C-HYK cow rumens was observed, suggesting these cows may be less efficient at degrading cellulose although the mechanistic role of Ruminococcus spp. in rumen metabolism is not completely understood. Substantial differences in fecal or rumen microbiomes among cows experiencing different levels of energy deficit were not observed, suggesting that hyperketonemia may not be greatly influenced by gut microbial composition, and vice versa. Further studies using higher resolution -omics approaches like meta-transcriptomics or meta-proteomics are needed to decipher the exact mechanisms at play.

Activated autophagy-lysosomal pathway in dairy cows with hyperketonemia is associated with lipolysis of adipose tissues

Activated autophagy-lysosomal pathway (ALP) can degrade virtually all kinds of cellular components, including intracellular lipid droplets, especially during catabolic conditions. Sustained lipolysis and increased plasma fatty acids concentrations are characteristic of dairy cows with hyperketonemia. However, the status of ALP in adipose tissue during this physiological condition is not well known. The present study aimed to ascertain whether lipolysis is associated with activation of ALP in adipose tissues of dairy cows with hyperketonemia and in calf adipocytes. In vivo, blood and subcutaneous adipose tissue (SAT) biopsies were collected from nonhyperketonemic (nonHYK) cows [blood β-hydroxybutyrate (BHB) concentration <1.2 mM, n = 10] and hyperketonemic (HYK) cows (blood BHB concentration 1.2-3.0 mM, n = 10) with similar days in milk (range: 3-9) and parity (range: 2-4). In vitro, calf adipocytes isolated from 5 healthy Holstein calves (1 d old, female, 30-40 kg) were differentiated and used for (1) treatment with lipolysis inducer isoproterenol (ISO, 10 µM, 3 h) or mammalian target of rapamycin inhibitor Torin1 (250 nM, 3 h), and (2) pretreatment with or without the ALP inhibitor leupeptin (10 μg/mL, 4 h) followed by ISO (10 µM, 3 h) treatment. Compared with nonHYK cows, serum concentration of free fatty acids was greater and serum glucose concentration, DMI, and milk yield were lower in HYK cows. In SAT of HYK cows, ratio of phosphorylated hormone-sensitive lipase to hormone-sensitive lipase, and protein abundance of adipose triacylglycerol lipase were greater, but protein abundance of perilipin 1 (PLIN1) and cell death-inducing DNA fragmentation factor-α-like effector c (CIDEC) was lower. In addition, mRNA abundance of autophagy-related 5 (ATG5), autophagy-related 7 (ATG7), and microtubule-associated protein 1 light chain 3 beta (MAP1LC3B), protein abundance of lysosome-associated membrane protein 1, and cathepsin D, and activity of β-N-acetylglucosaminidase were greater, whereas protein abundance of sequestosome-1 (p62) was lower in SAT of HYK cows. In calf adipocytes, treatment with ISO or Torin1 decreased protein abundance of PLIN1, and CIDEC, and triacylglycerol content in calf adipocytes, but increased glycerol content in the supernatant of calf adipocytes. Moreover, the mRNA abundance of ATG5, ATG7, and MAP1LC3B was upregulated, the protein abundance of lysosome-associated membrane protein 1, cathepsin D, and activity of β-N-acetylglucosaminidase were increased, whereas the protein abundance of p62 was decreased in calf adipocytes treated with ISO or Torin1 compared with control group. Compared with treatment with ISO alone, the protein abundance of p62, PLIN1, and CIDEC, and triacylglycerol content in calf adipocytes were higher, but the glycerol content in the supernatant of calf adipocytes was lower in ISO and leupeptin co-treated group. Overall, these data indicated that activated ALP is associated with increased lipolysis in adipose tissues of dairy cows with hyperketonemia and in calf adipocytes.

Effects of Exogenous Ketone Supplementation on Blood Glucose: A Systematic Review and Meta-analysis.

Recently developed ketone (monoester or salt) supplements acutely elevate blood β-hydroxybutyrate (BHB) exogenously without prolonged periods of fasting or carbohydrate restriction. Previous (small-scale) studies have found a blood glucose-lowering effect of exogenous ketones. This study aimed to systematically review available evidence and conduct meta-analyses of studies reporting on exogenous ketones and blood glucose. We searched 6 electronic databases on 13 December 2021 for randomized and nonrandomized trials of any length that reported on the use of exogenous ketones. We calculated raw mean differences (MDs) in blood BHB and glucose in 2 main analyses: 1) after compared with before acute ingestion of exogenous ketones and 2) following acute ingestion of exogenous ketones compared with a comparator supplement. We pooled effect sizes using random-effects models and performed prespecified subgroup analyses to examine the effect of potential explanatory factors, including study population, exercise, blood BHB, and supplement type, dosing, and timing. Risk of bias was examined using Cochrane's risk-of-bias tools. Studies that could not be meta-analyzed were summarized narratively. Forty-three trials including 586 participants are summarized in this review. Following ingestion, exogenous ketones increased blood BHB (MD=1.73 mM; 95% CI: 1.26, 2.21mM; P <0.001) and decreased mean blood glucose (MD = -0.54mM; 95% CI: -0.68, -0.40mM; P <0.001). Similarly, when compared with placebo, blood BHB increased (MD=1.98mM; 95% CI: 1.52, 2.45mM; P <0.001) and blood glucose decreased (MD = -0.47mM; 95% CI: -0.57, -0.36mM; P <0.001). Across both analyses, significantly greater effects were seen with ketone monoesters compared with salts (P <0.001). The available evidence indicates that acute ingestion of exogenous ketones leads to increased blood BHB and decreased blood glucose. Limited evidence on prolonged ketone supplementation was found.

Reducing milking frequency from twice to once daily as an adjunct treatment for ketosis in lactating dairy cows—A randomized controlled trial

This randomized controlled trial investigated the effects of temporarily reducing milking frequency (MF) on the resolution of ketosis and milk production in dairy cows in early lactation. To detect ketosis [blood β-hydroxybutyrate (BHB) ≥1.2 mmol/L], Holstein cows were screened daily from 3 to 16 d in milk using a cow-side meter. Cows diagnosed with ketosis (n = 104) were randomly assigned to twice-daily milking (TDM) or reduced to once-daily milking (ODM) for 2 wk, then returned to twice-daily milking. Both treatment groups received a 5-d treatment of an oral propylene glycol drench (PG; 300 g) beginning on the afternoon of the diagnosis; cows received additional 5-d PG treatments if they had a ketotic test result (blood BHB ≥1.2 mmol/L) at least 4 d after finishing the first PG treatment. Blood BHB tests were conducted for the first 3 d after ketosis diagnosis, and then once every 3 d for 21 d of trial (DOT). Milk and milk component data were collected weekly for 15 wk following trial enrollment. The ODM group showed rapidly and markedly decreased blood BHB concentrations (primiparous cows: 1 DOT, 0.92 ODM vs. 1.22 TDM, 15 DOT, 0.55 vs. 0.81 mmol/L; multiparous cows: 1 DOT, 1.01 vs. 1.40, 15 DOT, 0.78 vs. 1.65 mmol/L). In addition, a logistic regression model indicated that ODM cows were less likely to have blood BHB concentrations ≥1.2 mmol/L [primiparous cows: 3 DOT: ODM 1% (95% confidence interval: 0-10%) vs. TDM 43% (30-58%), 15 DOT ODM 0% (0-0.2%) vs. TDM 22% (13-36%); multiparous cows: 3 DOT: ODM 33% (24-44%) vs. TDM 59% (48-69%), 15 DOT ODM 20.9% (13-31%) vs. TDM 64% (53-74%)]. The proportion of ODM cows that required additional treatments of PG were substantially lower than the TDM group (ODM: 39%; TDM: 64%) than the TDM cows during the initial 21-d period. However, during the 2-wk treatment period, cows in the ODM group produced 26% less milk and 25% less energy-corrected milk than the TDM cows. During wk 3 to 15, when all cows were milked twice daily, ODM cows produced less milk (-14%) and energy-corrected milk (-12%) compared with the TDM group. Milk protein percentage was greater, and milk fat percentage and linear score tended to be greater in the ODM group over 15 wk. In conclusion, a 2-wk reduction of MF in ketotic cows from twice to once daily with treatment with PG resolved ketosis and decreased blood BHB concentrations more effectively than treating TDM cows with PG alone. However, the 2-wk MF reduction had immediate and long-term (up to 13 wk after cessation of MF reduction) negative effects on milk production.

Effects of postpartum milking strategy on plasma mineral concentrations and colostrum, transition milk, and milk yield and composition in multiparous dairy cows

The effects of postpartum milking strategy on plasma mineral concentrations, blood β-hydroxybutyrate (BHB) concentration, and colostrum, transition milk, and first monthly test milk yield and composition were evaluated in 90 multiparous Jersey and Jersey × Holstein crossbreed cows from a commercial farm. Before first postpartum milking, cows were randomly assigned to the following milking strategies, implemented during the first 2 d postpartum: twice-a-day milking (M2, standard industry practice, milking every 12 h; n = 22), once-a-day milking (M1, milking every 24 h; n = 24), restricted milking (MR, 3-L milking every 12 h; n = 21), and delayed milking (MD, no milking for the first 24 h, and milking every 12 h afterward; n = 23). Blood samples for total plasma Ca, P, and Mg determination were collected from enrollment every 4 h up to 48 h, and at 3 d in milk. Blood BHB concentration was determined at 3 and 11 d in milk. Colostrum and transition milk yields were recorded, and samples were collected at each study milking for IgG and somatic cell count (SCC) determinations. Information for first monthly test milk yield and composition was obtained from the Dairy Herd Improvement Association. Statistical analyses were conducted using generalized multiple linear and Poisson regressions with Dunnett adjustment and M2 as reference group for mean comparisons. Overall, plasma Ca concentration within 48 h after enrollment was higher for MD (2.17 mmol/L), tended to be higher for MR (2.15 mmol/L), and was similar for M1 (2.09 mmol/L) compared with M2 cows (2.06 mmol/L). No statistically significant differences compared with M2 cows were observed for plasma P and Mg concentrations. Colostrum and transition milk and total Ca harvested within 48 h after enrollment were lower for M1, MR, and MD compared with M2 cows. The MD strategy prevented harvesting colostrum with >50 g of IgG/L. No statistically significant effects were detected on plasma mineral concentrations at 3 DIM, blood BHB concentration, colostrum and transition milk SCC within 48 h after enrollment, or milk yield, energy-corrected milk yield, and SCC at first monthly test. Our results suggest that postpartum plasma Ca concentration may be influenced by postpartum milking strategy, without interfering with future milk yield and udder health. Further studies should evaluate whether the proposed milking strategies in early postpartum affect production, reproduction, or health.

Hepatic autophagy and mitophagy status in dairy cows with subclinical and clinical ketosis

Severe negative energy balance around parturition is an important contributor to ketosis, a metabolic disorder that occurs most frequently in the peripartal period. Autophagy and mitophagy are important processes responsible for breaking down useless or toxic cellular material, and in particular damaged mitochondria. However, the role of autophagy and mitophagy during the occurrence and development of ketosis is unclear. The objective of this study was to investigate autophagy and mitophagy in the livers of cows with subclinical ketosis (SCK) and clinical ketosis (CK). We assessed autophagy by measuring the protein abundance of microtubule-associated protein 1 light chain 3-II (LC3-II; encoded by MAP1LC3) and sequestosome-1 (p62, encoded by SQSTM1), as well as the mRNA abundance of autophagy-related genes 5 (ATG5), 7 (ATG7), and 12 (ATG12), beclin1 (BECN1), and phosphatidylinositol 3-kinase catalytic subunit type 3 (PIK3C3). Mitophagy was evaluated by measuring the protein abundance of the mitophagy upstream regulators PTEN-induced putative kinase 1 (PINK1) and Parkin. Liver and blood samples were collected from healthy cows [n = 15; blood β-hydroxybutyrate (BHB) concentration <1.2 mM], cows with SCK (n = 15; blood BHB concentration 1.2 to 3.0 mM) and cows with CK (n = 15; blood BHB concentration >3.0 mM with clinical signs) with similar lactation numbers (median = 3, range = 2 to 4) and days in milk (median = 6, range = 3 to 9). The serum activity of aspartate aminotransferase and alanine aminotransferase was greater in cows with CK than in healthy cows. Levels of oxidative stress biomarkers malondialdehyde and hydrogen peroxide were also higher in liver tissue from ketotic cows (SCK and CK) than from healthy cows. Compared with cows with CK and healthy cows, the hepatic mRNA abundance of MAP1LC3, SQSTM1, ATG5, ATG7, ATG12, and PIK3C3 was upregulated in cows with SCK. Compared with healthy cows, cows with SCK had a lower abundance of p62 and a greater abundance of LC3-II, but levels of both were higher in cows with CK. The mRNA abundance of ATG12 was lower in cows with CK than in healthy cows. Furthermore, the hepatic protein abundance of PINK1 and Parkin was greater in cows with SCK and slightly lower in cows with CK than in healthy cows. These data demonstrated differences in the hepatic activities of autophagy and mitophagy in cows with SCK compared with cows with CK. Although the precise mechanisms for these differences could not be discerned, autophagy and mitophagy seem to be involved in ketosis.

The Early Response to Dietary Therapy can Predict the Late Outcome in Children with Intractable Epilepsy.

Background and PurposeDietary therapy (DT), including the ketogenic diet (KD), is one of the nonpharmacological treatment options for patient with drug-resistant epilepsy. However, maintaining DT in patients without seizure reduction is very difficult, so it is critical for clinicians to decide when to stop this intervention.MethodsWe retrospectively analyzed early clinical and laboratory findings and the clinical characteristics of children who received DT. The maintenance of DT and the clinical seizure frequency were assessed at 1, 3, 6, 12, and 24 months after KD initiation. Responders were defined as patients showing an overall reduction in seizure frequency of >50% relative to the baseline.ResultsWe included 67 patients who received DT, but only 23 (34.3%) of these patients remained on DT at 6 months. Only 1 (5%) of the 20 responders at 1 month became a nonresponder at 6 months. The response rate at 6 months was significantly higher among patients under 2 years of age (15/17, 88.2%) than older patients (2/6, 33.3%; p=0.021). Moreover, the 6-month responders were significantly younger (29.4±38.6 months, mean±SD) than the nonresponders (98.9±84.6 months, p=0.012) at the initiation of the diet. A high blood β-hydroxybutyrate (BHB) level at 1 month predicted a good DT response at 6 months.ConclusionsMost 1-month responders maintained their response on DT for up to 6 months. The blood BHB level at 1 month was significantly correlated with the 6-month seizure outcome. Confirming clinical and laboratory biomarkers for the efficacy of DT requires further studies with larger cohorts.

Heritability estimates of predicted blood β-hydroxybutyrate and nonesterified fatty acids and relationships with milk traits in early-lactation Holstein cows

At the beginning of lactation, high-producing cows commonly experience an unbalanced energy status that is often responsible for the onset of metabolic disorders and impaired health and performance. Blood β-hydroxybutyrate (BHB) and nonesterified fatty acids (NEFA) are indicators of excessive fat mobilization and circulating ketone bodies. Recently, prediction models based on mid-infrared (MIR) spectroscopy have been developed to assess blood BHB and NEFA from routinely collected individual milk samples. This study aimed to estimate genetic parameters of blood BHB and NEFA predicted from milk MIR spectra and to assess their phenotypic and genetic correlations with milk production and composition traits in early-lactation Holstein cows. The data set comprised the first test-day record within lactation and spectra of individual milk samples (n = 22,718) of 13,106 Holstein cows collected from 5 to 35 d in milk (DIM). Blood BHB and NEFA were predicted from milk MIR spectra using previously developed prediction models. Genetic parameters of blood metabolites and milk traits were estimated for the whole observational period (5-35 DIM) and within 6 classes of DIM. Blood BHB and NEFA showed similar genetic variation across DIM, with the highest heritability in the first 10 d after calving (0.31 ± 0.06 and 0.19 ± 0.05 for BHB and NEFA, respectively). The genetic correlation between BHB and NEFA was moderate (0.51 ± 0.05). Genetic correlations of BHB with milk yield, SCS, protein percentage, lactose percentage, and urea nitrogen content were similar to, or at least in the same direction as, the correlations of NEFA with the same traits, whereas opposite correlations were observed with fat percentage and fat-to-protein ratio. Results of the current study suggest that blood BHB and NEFA predicted from milk MIR spectra have genetic variation that is potentially exploitable for breeding purposes. Therefore, they could be used as indicator traits of hyperketonemia in a selection index aimed to reduce the susceptibility of dairy cows to metabolic disorders in early lactation.

Variation of Blood Metabolites of Brown Swiss, Holstein-Friesian, and Simmental Cows.

Simple SummaryPopulation-level phenotyping of blood metabolites is hardly achievable due to the limitation of reference analyses. Mid-infrared spectroscopy has recently been used to develop prediction models for major blood metabolites, allowing their determination on a large scale. The current study investigated the variation of blood β-hydroxybutyrate, non-esterified fatty acids, and urea nitrogen predicted from a large milk mid-infrared spectra database of Brown Swiss, Holstein-Friesian, and Simmental cows. Holstein-Friesian cows had the greatest concentrations of β-hydroxybutyrate and non-esterified fatty acids, and the lowest urea nitrogen in blood, which may underline an altered energy and nutritional status.Serum metabolic profile is a common method to monitor health and nutritional status of dairy cows, but blood sampling and analysis are invasive, time-consuming, and expensive. Milk mid-infrared spectra have recently been used to develop prediction models for blood metabolites. The current study aimed to investigate factors affecting blood β-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), and urea nitrogen (BUN) predicted from a large milk mid-infrared spectra database. Data consisted of the first test-day record of early-lactation cows in multi-breed herds. Holstein-Friesian cows had the greatest concentration of blood BHB and NEFA, followed by Simmental and Brown Swiss. The greatest and the lowest concentrations of BUN were detected for Brown Swiss and Holstein-Friesian, respectively. The greatest BHB concentration was observed in the first two weeks of lactation for Brown Swiss and Holstein-Friesian. Across the first month of lactation, NEFA decreased and BUN increased for all considered breeds. The greatest concentrations of blood BHB and NEFA were recorded in spring and early summer, whereas BUN peaked in December. Environmental effects identified in the present study can be included as adjusting factors in within-breed estimation of genetic parameters for major blood metabolites.

Blood β-hydroxybutyrate concentrations and early lactation management strategies on pasture-based dairy farms in Colombia

The increasing global demand for food requires sustainable solutions to close the gap in agricultural yield between industrialized and non-industrialized countries. Our objectives in this cross-sectional study were to: 1) characterize farm populations, milk yield, and early lactation management strategies of dairy cows in three different regions of Colombia, and 2) determine the association of these management strategies with blood β-hydroxybutyrate (BHB) concentrations in the first 42 days in milk (DIM). Dairy herds (n = 56) in the Antioquia, Caldas, and Cundinamarca regions of Colombia were visited once from May through July 2018. A survey was administered to farm owners to collect demographic, management, and herd nutrition information. Blood samples from dairy cows (n = 880) between calving and 42 DIM were used to measure blood BHB concentration. Associations between management and nutritional strategies and blood BHB concentration were examined using mixed models. Prevalence of hyperketonemia was calculated as the number of samples with BHB concentration ≥1.2 mmol/L divided by the total number of samples. The estimated diet composition for early lactation dairy cows was 65.5% pasture and 31.8% commercial concentrates. The farm median milk yield, protein concentration, and fat concentration were 21.0 kg (range = 13.1–36 kg), 3.2% (range = 2.7–4.1%), and 3.5% (range = 3.0–4.1%), respectively. Milk yield least squares means (95% confidence interval; CI) differed by region: 21.7 (20.3, 23.2), 18.5 (17.0, 20.2), and 20.3 (18.5, 22.4) kg in Antioquia, Caldas, and Cundinamarca, respectively. Median blood BHB concentration was 0.5 and ranged from 0.1–4.4 mmol/L; blood BHB concentration was not different among the three regions. Pasture fertilization, increased parity, and BCS were associated with changes in blood BHB concentration. The overall prevalence of hyperketonemia was 4.5%. Geographical region affected the prevalence of hyperketonemia at 2.5%, 4.0%, and 10.2% in Antioquia, Caldas, and Cundinamarca, respectively. Mean stocking density (95% CI) was greater in Cundinamarca than Antioquia or Caldas at 3.3 (2.2, 5.0), 2.8 (2.1, 3.9) and 1.7 (1.2, 2.6) animals per ha, respectively, and was associated with hyperketonemia prevalence. Farms that abruptly stop milking cows at dry-off had 80% of the hyperketonemia events in the study. Pasture-based dairies in Colombia had lower blood BHB concentrations and estimated milk yield compared with confined production systems in temperate zones. However, geographical region, stocking density, and abrupt cessation of milking at dry-off were associated with prevalence of hyperketonemia in pasture-based dairies.

Prediction of blood metabolites from milk mid-infrared spectra in early-lactation cows

Dairy cows commonly experience an unbalanced energy status in early lactation, and this condition can lead to the onset of several metabolic disorders. Blood metabolic profile testing is a valid tool to monitor and detect the most common early lactation disorders, but blood sampling and analysis are time-consuming and expensive, and the procedure is invasive and stressful for the cows. Mid-infrared (MIR) spectroscopy is routinely used to analyze milk composition, being a cost-effective and nondestructive method. The present study aimed to assess the feasibility of using routine milk MIR spectra for the prediction of main blood metabolites in dairy cows, and to investigate associations between measured blood metabolites and milk traits. Twenty herds of Holstein Friesian, Brown Swiss, or Simmental cows located in Northeast Italy were visited 1 to 4 times between December 2017 and June 2018, and blood and milk samples were collected from all lactating cows within 35 d in milk. Concentrations of main blood metabolites and milk MIR spectra were recorded from 295 blood and milk samples and used to develop prediction models for blood metabolic traits through backward interval partial least squares analysis. Blood β-hydroxybutyrate (BHB), urea, and nonesterified fatty acids were the most predictable traits, with coefficients of determination of 0.63, 0.58, and 0.52, respectively. On the contrary, predictive performance for blood glucose, triglycerides, cholesterol, glutamic oxaloacetic transaminase, and glutamic pyruvic transaminase were not accurate. Associations of blood BHB and urea with their respective contents in milk were moderate to strong, whereas all other correlations were weak. Predicted blood BHB showed an improved performance in detecting cows with hyperketonemia (blood BHB ≥ 1.2 mmol/L), compared with commercial calibration equation for milk BHB. Results highlighted the opportunity of using milk MIR spectra to predict blood metabolites and thus to collect routine information on the metabolic status of early-lactation cows at a population level.

Prediction of blood β-hydroxybutyrate content and occurrence of hyperketonemia in early-lactation, pasture-grazed dairy cows using milk infrared spectra

The objective of this study was to evaluate the ability of milk infrared spectra to predict blood β-hydroxybutyrate (BHB) concentration for use as a management tool for cow metabolic health on pasture-grazed dairy farms and for large-scale phenotyping for genetic evaluation purposes. The study involved 542 cows (Holstein-Friesian and Holstein-Friesian × Jersey crossbreds), from 2 farms located in the Waikato and Taranaki regions of New Zealand that operated under a seasonal-calving, pasture-based dairy system. Milk infrared spectra were collected once a week during the first 5 wk of lactation. A blood "prick" sample was taken from the ventral labial vein of each cow 3 times a week for the first 5 wk of lactation. The content of BHB in blood was measured immediately using a handheld device. After outlier elimination, 1,910 spectra records and corresponding BHB measures were used for prediction model development. Partial least square regression and partial least squares discriminant analysis were used to develop prediction models for quantitative determination of blood BHB content and for identifying cows with hyperketonemia (HYK). Both quantitative and discriminant predictions were developed using the phenotypes and infrared spectra from two-thirds of the cows (randomly assigned to the calibration set) and tested using the remaining one-third (validation set). A moderate accuracy was obtained for prediction of blood BHB. The coefficient of determination (R2) of the prediction model in calibration was 0.56, with a root mean squared error of prediction of 0.28 mmol/L and a ratio of performance to deviation, calculated as the ratio of the standard deviation of the partial least squares model calibration set to the standard error of prediction, of 1.50. In the validation set, the R2 was 0.50, with root mean squared error of prediction values of 0.32 mmol/L, which resulted in a ratio of performance to deviation of 1.39. When the reference test for HYK was defined as blood concentration of BHB ≥1.2 mmol/L, discriminant models indicated that milk infrared spectra correctly classified 76% of the HYK-positive cows and 82% of the HYK-negative cows. The quantitative models were not able to provide accurate estimates, but they could differentiate between high and low BHB concentrations. Furthermore, the discriminant models allowed the classification of cows with reasonable accuracy. This study indicates that the prediction of blood BHB content or occurrence of HYK from milk spectra is possible with moderate accuracy in pasture-grazed cows and could be used during routine milk testing. Applicability of infrared spectroscopy is not likely suited for obtaining accurate BHB measurements at an individual cow level, but discriminant models might be used in the future as herd-level management tools for classification of cows that are at risk of HYK, whereas quantitative models might provide large-scale phenotypes to be used as an indicator trait for breeding cows with improved metabolic health.

Genetic variation in serum protein pattern and blood β-hydroxybutyrate and their relationships with udder health traits, protein profile, and cheese-making properties in Holstein cows

The aim of this study was to investigate in Holstein cows the genetic basis of blood serum metabolites [i.e., total protein, albumin, globulin, albumin:globulin ratio (A:G), and blood β-hydroxybutyrate (BHB)], a set of milk phenotypes related to udder health, milk quality technological characteristics, and genetic relationships among them. Samples of milk were collected from 498 Holstein cows belonging to 28 herds. All animal welfare and milk phenotypes were assessed using standard analytical methodology. A set of Bayesian univariate and bivariate animal models was implemented via Gibbs sampling, and statistical inference was based on the marginal posterior distributions of parameters of concern. We observed a small additive genetic influence for serum albumin concentrations, moderate heritability (≥0.20) for total proteins, globulins, and A:G, and high heritability (0.37) for blood BHB. Udder health traits (somatic cell score, milk lactose, and milk pH) showed low or moderate heritabilities (0.15-0.20), whereas variations in milk protein fraction concentrations were confirmed as mostly under genetic control (heritability: 0.21-0.71). The moderate and high heritabilities observed for milk coagulation properties and curd firming modeling parameters provided confirmation that genetic background exerts a strong influence on the cheese-making ability of milk, largely due to genetic polymorphisms in the major milk protein genes. Blood BHB showed strong negative genetic correlations with globulins (-0.619) but positive correlations with serum albumin (0.629) and A:G (0.717), which suggests that alterations in the serum protein pattern and BHB blood levels are likely to be genetically related. Strong relationships were found between albumin and fat percentages (-0.894), between globulin and αS2-CN (-0.610), and, to a lesser extent, between serum protein pattern and milk technological characteristics. Genetic relationships between blood BHB and traits related to udder health and milk quality and technological characteristics were mostly weak. This study provides evidence that there is exploitable additive genetic variation for traits related to animal health and welfare and throws light on the shared genetic basis of these traits and the phenotypes related to the quality and cheese-making ability of milk.

Milk yield relative to supplement intake and rumination time differs by health status for fresh cows milked with automated systems

Automated milking systems (AMS) provide the capability of adjusting the amount of supplemental concentrate offered to cows based on parity, days in milk, and milk yield; in many situations, more supplement is provided to those cows with higher production demands. To determine associations of subclinical ketosis (SCK) and blood β-hydroxybutyrate (BHB) with milk yield, supplement consumed in the AMS, rumination time, and the ratios of milk yield to supplement intake and milk yield to rumination time, we monitored 605 cows from 9 AMS herds, testing blood BHB concentrations 1×/wk for the first 3 wk of lactation. Milk yield, supplement intake, and rumination data were collected from the AMS computer on each farm. For analyses, only multiparous cows from 8 herds were included (n = 172 total) and were matched, within farm, to include an equal number of cows in each of 4 health status groups (n = 43 per group): SCK (BHB ≥1.2 mmol/L at ≥1 of 3 tests, with no other disorder in the first 30 d in milk), SCK+ (BHB ≥1.2 mmol/L at ≥1 of 3 tests, with another health disorder), HLT (BHB always <1.2 mmol/L, with no other disorder), or OTH (BHB always <1.2 mmol/L, with a health disorder). Milk yield and rumination time varied by health status, but we found no difference in supplement intake by health status group. As a result, milk yield relative to supplement intake and milk yield relative to rumination time differed by health status and were both positively associated with BHB. Cows in SCK had the highest milk yield and ratio of milk yield to supplement intake, but SCK+ cows had the lowest rumination time and the highest milk yield to rumination ratio. These results highlight the differences in milk production (per day and relative to supplement consumption or rumination time) associated with blood BHB and health status. Therefore, potential exists to modify supplementation in AMS to reduce negative energy balance by accounting for milk production of fresh cows.

Prepartal standing behavior as a parameter for early detection of postpartal subclinical ketosis associated with inflammation and liver function biomarkers in peripartal dairy cows

A degree of negative energy balance is commonly experienced by cows during early lactation. This physiological state, if pronounced or prolonged, leads to partial oxidation of nonesterified fatty acids as an energy source and, consequently, increasing blood β-hydroxybutyrate (BHB) concentrations and potentially development of ketosis in postpartal dairy cows. Twenty-four multiparous Holstein cows received a common prepartal and postpartal diet. Cows were fitted with an accelerometer mounted laterally on the distal left hind leg using vet wrap from -30 to 15 d relative to parturition. A retrospective analysis was performed using the postpartal BHB data at 8 time points from 0 to 15 d in milk measured with the Precision Xtra (Abbott Diabetes Care, Alameda, CA). Cows with an average blood BHB <1.4 mmol/L were designated nonketotic (NONKET; n = 12), and those with ≥1.4 mmol/L were designated ketotic (KET; n = 12). A total of 8 samples per cow were used for this analysis. Subsequent analyses of behavioral patterns and blood biomarkers were performed using this group effect. On average, blood BHB reached subclinical levels (1.4 ± 0.3 mmol/L; mean ± standard error of the mean) at 3 d postpartum for all cows in this study. Behavioral patterns were obtained from accelerometer data, and correlation analysis was performed between these behaviors such as standing and lying time from -30 to 3 d relative to parturition and blood BHB concentration at 3 d postpartum. The strongest correlation was obtained between standing time at 3 d before calving and blood BHB at 3 d postpartum. Dry matter intake was greater (ca. 3 kg/d) in NONKET cows than in KET cows. An interaction of group × time for milk yield resulted in an overall increase of 5.7 kg/d in NONKET cows in comparison with KET. The blood concentrations of biomarkers for liver function (γ-glutamyltransferase and glutamic-oxaloacetic transaminase), inflammation (IL-6), and metabolism (nonesterified fatty acids) were increased at various time points in KET cows in comparison with NONKET during the transition period. Overall, lower bilirubin in NONKET cows than in KET further confirmed an impaired liver function in the latter group of cows. Our findings revealed the potential for establishing correlations between prepartal behavioral patterns derived from accelerometer data and postpartal subclinical ketosis, and further confirming the latter by physiological alterations in biomarkers related to inflammation and liver function. Our data also indicate that cows with a predisposition to postpartal subclinical or clinical ketosis will remain standing for fewer hours during the days leading to parturition, which decreased DMI, and this condition was further reflected in lower milk yield.

Predicting blood β-hydroxybutyrate using milk Fourier transform infrared spectrum, milk composition, and producer-reported variables with multiple linear regression, partial least squares regression, and artificial neural network

Prediction of postpartum hyperketonemia (HYK) using Fourier transform infrared (FTIR) spectrometry analysis could be a practical diagnostic option for farms because these data are now available from routine milk analysis during Dairy Herd Improvement testing. The objectives of this study were to (1) develop and evaluate blood β-hydroxybutyrate (BHB) prediction models using multivariate linear regression (MLR), partial least squares regression (PLS), and artificial neural network (ANN) methods and (2) evaluate whether milk FTIR spectrum (mFTIR)-based models are improved with the inclusion of test-day variables (mTest; milk composition and producer-reported data). Paired blood and milk samples were collected from multiparous cows 5 to 18 d postpartum at 3 Wisconsin farms (3,629 observations from 1,013 cows). Blood BHB concentration was determined by a Precision Xtra meter (Abbot Diabetes Care, Alameda, CA), and milk samples were analyzed by a privately owned laboratory (AgSource, Menomonie, WI) for components and FTIR spectrum absorbance. Producer-recorded variables were extracted from farm management software. A blood BHB ≥1.2 mmol/L was considered HYK. The data set was divided into a training set (n = 3,020) and an external testing set (n = 609). Model fitting was implemented with JMP 12 (SAS Institute, Cary, NC). A 5-fold cross-validation was performed on the training data set for the MLR, PLS, and ANN prediction methods, with square root of blood BHB as the dependent variable. Each method was fitted using 3 combinations of variables: mFTIR, mTest, or mTest + mFTIR variables. Models were evaluated based on coefficient of determination, root mean squared error, and area under the receiver operating characteristic curve. Four models (PLS-mTest + mFTIR, ANN-mFTIR, ANN-mTest, and ANN-mTest + mFTIR) were chosen for further evaluation in the testing set after fitting to the full training set. In the cross-validation analysis, model fit was greatest for ANN, followed by PLS and MLR. Diagnostic strength after cross-validation was poorest for MLR and was similar for ANN and PLS. Models that used mTest + mFTIR variables performed marginally better than models that used only mFTIR or mTest variables. These results suggest that blood BHB prediction models that use mFTIR + mTest variables may be useful additions to existing HYK diagnostic and management programs.