Effects of time and sampling location on concentrations of β-hydroxybutyric acid in dairy cows

Two trials were conducted to examine factors potentially influencing the measurement of blood β-hydroxybutyric acid (BHBA) in dairy cows. The objective of the first trial was to study effects of sampling time on BHBA concentration in continuously fed dairy cows. Furthermore, we determined test characteristics of a single BHBA measurement at a random time of the day to diagnose subclinical ketosis considering commonly used cut-points (1.2 and 1.4 mmol/L). Finally, we set out to evaluate if test characteristics could be enhanced by repeating measurements after different time intervals. During 4 herd visits, a total of 128 cows (8 to 28 d in milk) fed 10 times daily were screened at 0900 h and preselected by BHBA concentration. Blood samples were drawn from the tail vessels and BHBA concentrations were measured using an electronic BHBA meter (Precision Xceed, Abbott Diabetes Care Ltd., Witney, UK). Cows with BHBA concentrations ≥0.8 mmol/L at this time were enrolled in the trial (n = 92). Subsequent BHBA measurements took place every 3 h for a total of 8 measurements during 24 h. The effect of sampling time on BHBA concentrations was tested in a repeated-measures ANOVA repeating sampling time. Sampling time did not affect BHBA concentrations in continuously fed dairy cows. Defining the average daily BHBA concentration calculated from the 8 measurements as the gold standard, a single measurement at a random time of the day to diagnose subclinical ketosis had a sensitivity of 0.90 or 0.89 at the 2 BHBA cut-points (1.2 and 1.4 mmol/L). Specificity was 0.88 or 0.90 using the same cut-points. Repeating measurements after different time intervals improved test characteristics only slightly. In the second experiment, we compared BHBA concentrations of samples drawn from 3 different blood sampling locations (tail vessels, jugular vein, and mammary vein) of 116 lactating dairy cows. Concentrations of BHBA differed in samples from the 3 sampling locations. Mean BHBA concentration was 0.3 mmol/L lower when measured in the mammary vein compared with the jugular vein and 0.4 mmol/L lower in the mammary vein compared with the tail vessels. We conclude that to measure BHBA, blood samples of continuously fed dairy cows can be drawn at any time of the day. A single measurement provides very good test characteristics for on-farm conditions. Blood samples for BHBA measurement should be drawn from the jugular vein or tail vessels; the mammary vein should not be used for this purpose.     

Associations of elevated nonesterified fatty acids and β-hydroxybutyrate concentrations with early lactation reproductive performance and milk production in transition dairy cattle in the northeastern United States

The objectives were to evaluate the effects of elevated pre- and postpartum nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) concentrations during the transition period on reproductive performance and milk production in dairy cattle. In a prospective cohort study of 91 freestall, total mixed ration-fed herds in the northeastern United States, blood samples were collected from approximately 15 prepartum and 15 different postpartum transition animals in each herd. All samples were stratified based on pre- or postpartum status at the time of sample collection, and 2,259 and 2,290 animals were used to evaluate reproductive and milk production performance, respectively. Reproductive performance was assessed by time to conception within 70 d post-voluntary waiting period (VWP) and milk production was assessed using mature-equivalent 305-d (ME305) milk yield estimated at 120 d in milk. While controlling for body condition score (BCS), calving season, median ME305 milk production, and parity, NEFA and BHBA concentrations were evaluated with time to event analysis to investigate reproductive performance. These same predictor variables were used to determine the effects of elevated NEFA and BHBA concentrations on ME305 milk yield with herd as a random effect. Heifers and cows were grouped in the final analyses if the results between groups were similar. In all animals sampled prepartum, the risk of pregnancy within 70 d post-VWP was reduced by 19% when NEFA concentrations were ≥0.27 mEq/L. In all animals sampled postpartum, those with NEFA concentrations ≥0.72 mEq/L had a 16% decrease in risk of pregnancy and those with BHBA concentrations ≥10 mg/dL had a 13% decrease in risk. In cows and heifers, ME305 milk yield was decreased by 683 kg when prepartum NEFA concentrations were ≥0.33 mEq/L. In heifers sampled postpartum, ME305 milk yield was increased by 488 kg when NEFA concentrations were ≥0.57 mEq/L and increased by 403 kg when BHBA concentrations were ≥9 mg/dL. In cows sampled postpartum, ME305 milk yield was decreased by 647 kg when NEFA concentrations were ≥0.72 mEq/L and decreased by 393 kg when BHBA concentrations were ≥10 mg/dL. With the exception of milk production in heifers, this study indicates that increased concentrations of serum NEFA and BHBA had a detrimental effect on reproductive performance and milk production.     

Short communication: Oxidative status and incidence of mastitis relative to blood α-tocopherol concentrations in the postpartum period in dairy cows

Vitamin E supplementation, when combined with high blood α-tocopherol (>6.25 μg/mL) at dry off, has been reported to unexpectedly increased the risk for clinical mastitis in dairy cows. Furthermore, higher levels of oxidative stress in the postpartum period were related to higher risk of mastitis. The objective of the present study was to determine the relationship between various serum biomarkers of oxidative status, incidence of mastitis, and blood α-tocopherol concentrations at dry off and at calving. A total of 146 dairy cows from a commercial farm were used in an observational field study. All cows were supplemented with 3,000 and 50 IU/cow per day of all-rac-α-tocopherol during the dry period and lactation, respectively. Blood samples were collected at dry off and at calving. Serum was analyzed for α-tocopherol, levels of reactive oxygen metabolites (ROM), thiol groups (SH), and ferric-reducing ability. Three α-tocopherol groups at calving were created: high (>3 μg/mL), medium (2–3 μg/mL), and low (<2 μg/mL). Three α-tocopherol groups at dry off were created: high (>6.25 μg/mL), medium (4.25–6.25 μg/mL), and low (<4.25 μg/mL). All cases of clinical mastitis that occurred during the dry period and the entire subsequent lactation were verified by a veterinarian. No differences were observed in the incidence of mastitis between the 3 α-tocopherol groups based on the serum levels at dry off. Incidence of mastitis was 4 times lower in the high and medium groups when compared with the corresponding value for the low-α-tocopherol group based on the serum levels at calving. Lower levels of ROM and SH at dry off and at calving were found in the group of cows with the highest α-tocopherol values at dry off when compared with the corresponding values in the low-α-tocopherol group. The ROM values at dry off but not at calving were lower in the group of cows with the highest α-tocopherol values at calving when compared with the corresponding values in the low-α-tocopherol group. No differences were observed in ferric-reducing ability values between the 3 α-tocopherol groups at dry off or calving. No differences were observed in all biomarkers of oxidative status between healthy cows and those with mastitis. Thus, blood α-tocopherol is inversely related to certain biomarkers of oxidative stress in the postpartum period and incidence of mastitis. However, reduction in the incidence of mastitis is not mediated through a reduction in the levels of various biomarkers of oxidative stress.     

Short communication: Free fatty acid receptors FFAR1 and FFAR2 during the peripartal period in liver of dairy cows grouped by their postpartum plasma β-hydroxybutyrate concentrations

Free fatty acid receptors (FFAR) play significant roles in various physiological processes, including energy metabolism, through interaction with their ligands, fatty acids. To determine whether the receptors FFAR1 and FFAR2 are involved in the regulation of liver metabolism during the peripartal period, we selected 13 German Holstein multiparous dairy cows and grouped them as high β-hydroxybutyrate (H-BHB; n = 8) or low β-hydroxybutyrate (L-BHB; n = 5) according to their individual maximum plasma BHB concentration observed within wk 2 or 3 postpartum (H-BHB: >1 mmol/L and L-BHB: <0.77 mmol/L). The selected cows had a milk yield of more than 10,000 kg/305 d during a previous lactation. The cows were fed a total mixed ration according to their requirements during the far-off dry period [5.9 MJ of net energy for lactation (NE_L)/kg of dry matter (DM), crude protein (CP) 126 g/kg of DM], close-up dry period (6.5 MJ of NE_L/kg of DM, CP 137 g/kg of DM), and lactation (7 MJ of NE_L/kg of DM, CP 163 g/kg of DM). Blood samples were taken weekly, from d ?34 to d 40 relative to parturition. Liver biopsies were taken on d ?34, ?17, 3, 18, and 30 relative to parturition and at slaughter (d 40). The protein abundance of FFAR1 was lower during the whole peripartal period in the H-BHB group. The abundance of FFAR2 increased over time and tended to be higher in H-BHB cows. The abundance of FFAR1 might be associated with imbalances of liver metabolism in peripartal dairy cows.     

Evaluation of nonesterified fatty acids and β-hydroxybutyrate in transition dairy cattle in the northeastern United States: Critical thresholds for prediction of clinical diseases

The objectives of this study were to 1) establish cow-level critical thresholds for serum concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) to predict periparturient diseases [displaced abomasa (DA), clinical ketosis (CK), metritis and retained placenta, or any of these three], and 2) investigate the magnitude of the metabolites’ association with these diseases within 30 d in milk. In a prospective cohort study of 100 freestall, total mixed ration-fed herds in the northeastern United States, blood samples were collected from approximately 15 prepartum and 15 different postpartum transition animals in each herd, for a total of 2,758 samples. Serum NEFA concentrations were measured in the prepartum group, and both NEFA and BHBA were measured in the postpartum group. The critical thresholds for NEFA or BHBA were evaluated with receiver operator characteristic analysis for all diseases in both cohorts. The risk ratios (RR) of a disease outcome given NEFA or BHBA concentrations and other covariates were modeled with multivariable regression techniques, accounting for clustering of cows within herds. The NEFA critical threshold that predicted any of the 3 diseases in the prepartum cohort was 0.29 mEq/L and in the postpartum cohort was 0.57 mEq/L. The critical threshold for serum BHBA in the postpartum cohort was 10 mg/dL, which predicted any of the 3 diseases. All RR with NEFA as a predictor of disease were >1.8; however, RR were greatest in animals sampled postpartum (e.g., RR for DA = 9.7; 95% CI = 4.2 to 22.4. All RR with BHBA as the predictor of disease were >2.3 (e.g., RR for DA = 6.9; 95% CI = 3.7 to 12.9). Although prepartum NEFA and postpartum BHBA were both significantly associated with development of clinical disease, postpartum serum NEFA concentration was most associated with the risk of developing DA, CK, metritis, or retained placenta during the first 30 d in milk.     

Effect of vitamin E supplementation on α-tocopherol and β-carotene concentrations in tissues from pasture- and grain-fed cattle

The effects of dietary vitamin E supplementation of grain-fed cattle on lipid oxidation and meat colour have been extensively investigated, but little attention has been given to pasture-fed cattle where meat is likely to contain naturally high amounts of α-tocopherol and carotenoids. In the work described, we evaluated the effects of pasture-feeding alone and with vitamin E supplementation on tissue levels of anti-oxidants and compared the findings with those obtained for grain-fed cattle with and without supplementation. Sorghum was the major component of the grained-based ration. α-Tocopherol concentrations in plasma, muscle and fat tissues of pasture-fed cattle were not affected by vitamin E supplementation (2500 IU/head/day for 132 days prior to slaughter) while those of grain-fed cattle increased significantly. The α-tocopherol concentrations in the supplemented grain-fed cattle were similar in muscle and liver to pasture-fed animals but were lower in their fat (P<0.05). The major carotenoid present in all tissues studied from pasture-fed was β-carotene and its contents in plasma, liver, fat and muscles were decreased (P<0.05) by supplementation with vitamin E. Carotenoids were essentially absent in grain-fed cattle except for small amounts in liver. The implication of this study for the meat industry is that cattle grazed on good pasture can achieve concentrations of α-tocopherol in muscles and other tissues at least as high as those obtained by supra-nutritional supplementation of grain-fed cattle with vitamin E. However, α-tocopherol supplementation of pasture-fed cattle reduced tissue concentrations of β-carotene, which would reduce carcase fat yellowness and make pasture-fed cattle more acceptable to some Asian markets.     

Short communication: Relationships among plasma and milk vitamin B12, plasma free fatty acids,and blood β-hydroxybutyrate concentrations in early lactation dairy cows

This study was undertaken to evaluate the relationship between plasma and milk concentrations of vitamin B₁₂ as well as the relationship between plasma or milk concentrations of vitamin B₁₂ and plasma concentration of free fatty acids (FFA) or blood concentration of β-hydroxybutyrate (BHB) of early lactating Ayrshire (AY) and Holstein (HO) cows. A total of 44 dairy herds (7 AY and 37 HO herds) and 62 AY (21 in first, 19 in second, and 22 in third and more lactations) and 228 HO (51 in first, 74 in second, and 103 in third and more lactation) cows between 3 and 40 d in milk were involved in the study. Hand-stripped milk samples and blood samples were taken 6 h after the morning milking. Milk and plasma samples were analyzed for vitamin B₁₂ concentration and plasma samples were analyzed for FFA concentration. A handheld device was used for blood BHB concentration determination. Thresholds for elevated plasma FFA concentration and hyperketonemia were set at ≥0.70 and ≥1.2 mmol/L, respectively. Vitamin B₁₂ concentration in milk of AY primiparous cows [2,557 (1,995–3,276) pg/mL] was lower than in milk from HO primiparous cows [3,876 (3,356–4,478) pg/mL], whereas no difference was observed among other parities and breeds. Regardless of breeds, plasma concentration of vitamin B₁₂ of first and second parities was lower than plasma concentration of third and more lactation cows. Milk vitamin B₁₂ concentration was positively correlated with plasma vitamin B₁₂ concentration, but the relationship was stronger for AY (ρ averaging 0.63) than for HO cows (ρ averaging 0.36). For AY and HO breeds, a significant relationship between milk or plasma vitamin B₁₂ concentrations and plasma FFA concentration (ρ between 0.29 and 0.59) was observed. Moreover, cows with elevated plasma FFA concentration had greater milk and plasma vitamin B₁₂ concentrations than cows with normal plasma FFA concentration. No relationship between vitamin B₁₂ concentration in milk or plasma and blood BHB concentration and hyperketonemia was noted. In summary, milk is not well correlated with plasma vitamin B₁₂ concentration for HO. It could be hypothesized that elevated plasma concentration of FFA could have a negative effect on the use of vitamin B₁₂ by cow cells, which increases the concentration of the vitamin in plasma and its secretion in milk.     

Short communication: Responses to increasing amounts of free α-linolenic acid infused into the duodenum of lactating dairy cows

Increasing the α-linolenic acid (LNA; 18:3 cis-9,cis-12,cis-15) content of milk fat might help promote consumers’ health. The objective of this study was to determine the potential to alter the content of LNA in milk by duodenal infusion of a free fatty acid mixture rich in LNA. Four multiparous lactating Chinese Holstein cows fitted with duodenal cannulas were administered 2 treatments in a crossover design: an LNA-rich fatty acid infusion at varying concentrations (0, 40, 80, 120, and 160 g/d) versus a basal infusate control. Dry matter intake was not affected by LNA infusions. Milk production tended to decrease and was quadratically affected as LNA infusion increased, but 4% fat-corrected milk yield was not changed. Milk fat content tended to increase linearly with LNA infusion. Milk protein content was not changed by LNA infusion, whereas milk lactose content and yield were decreased quadratically as LNA infusion increased. Increasing the amount of LNA infused into the duodenum linearly increased concentrations of 18:3 cis-9,cis-12,cis-15 (0.61 to 25.4 g/100 g of total fatty acids) and 18:2 cis-9,cis-12 in milk fat. Increasing LNA decreased the percentages of 4:0, 14:0, and 16:0 fatty acids linearly. Increasing LNA also linearly decreased the percentages of 18:1 cis-9 and 18:2 cis-9,trans-11 in milk fat. Milk fat content of 20:5 cis-5,cis-8,cis-11,cis-14,cis-17 was quadratically affected, whereas concentrations of 18:0, 18:1 trans-9, 18:1 trans-11, and 18:2 trans-10,cis-12 were not affected. Increasing the supply of 18:3 cis-9,cis-12,cis-15 to the small intestine linearly increased 18:3 cis-9,cis-12,cis-15 in milk fat and markedly altered milk fat composition.     

The effect of injectable butaphosphan and cyanocobalamin on postpartum serum β-hydroxybutyrate, calcium, and phosphorus concentrations in dairy cattle

The objective of this study was to determine the effect of an injection of 10% butaphosphan and cyanocobalamin (Catosal, Bayer, Shawnee Mission, KS) on the day of calving and 1 d later on the prevalence of subclinical ketosis in dairy cattle in the early postpartum period. Cows from 4 herds (n = 1,122) were randomized to receive either 25 mL of 10% butaphosphan and cyanocobalamin or 25 mL of sterile water subcutaneously on both days. Each milliliter of Catosal contained 0.05 mg of cyanocobalamin and 100 mg of butaphosphan, which provided 17.3 mg of P in the form of [1-(butylamino)-1-methylethyl]-phosphonic acid. Serum was sampled for β-hydroxybutyrate (BHBA) concentration at calving (pretreatment) and again between 3 and 10 d in milk. A subset of samples from mature cows was also evaluated for serum Ca and P concentrations. When cows from all age groups were included in the analysis, there was no difference between the median serum BHBA concentrations of cows in the 2 treatment groups, and no difference in the proportion of hyperketonemic cows (serum BHBA ≥1,200 μmol/L) during the first week postpartum. When the analysis was restricted to mature cows (lactation ≥3), both the median BHBA concentration and the proportion of hyperketonemic cows were significantly lower in the treatment group than in the placebo group. Serum Ca and P concentrations did not differ between treatment groups. Our results suggest that injection of butaphosphan and cyanocobalamin on the day of calving and 1 d later may decrease the prevalence of subclinical ketosis during the week after calving in mature dairy cows, but not in first- and second-lactation animals.     

The effect of different treatments for early-lactation hyperketonemia on blood β-hydroxybutyrate,plasma nonesterified fatty acids,glucose, insulin,and glucagon in dairy cattle

Despite increased efforts in preventing the occurrence of metabolic disorders in transition cows, hyperketonemia remains a frequent early-lactation metabolic disease affecting an average of 40% of cows in herds in the United States. Despite the demonstrated economic effect of this disorder, controlled clinical trials comparing different treatment strategies in affected cows are lacking. The objective of our study was to investigate the effect of treatment with intravenous glucose, oral propylene glycol, or a combination of both on the reduction in blood β-hydroxybutyrate (BHB) concentrations of early-lactation hyperketonemic dairy cows. Multiparous Holstein cows between 3 to 9 d in milk were screened for hyperketonemia using a handheld meter 3 times per week, and enrolled at whole blood BHB concentration ≥1.2 mmol/L to 1 of 4 treatment groups: (1) 500 mL of a 50% dextrose solution i.v. once daily for 3 d (GLU, n = 9), (2) 300 mL of propylene glycol as a drench once daily for 3 d (PG, n = 9), (3) a combination treatment of a 500 mL of 50% dextrose solution i.v. and 300 mL of propylene glycol orally once daily for 3 d (GLU+PG, n = 8), or (4) an untreated control group (CTRL, n = 8). Blood samples were collected immediately before as well as at 1, 2, 4, 8, 12, 24, 36, 48, 60, and 72 h after administration of the first treatment through a jugular catheter and 3 times per week thereafter from coccygeal vessels. Concentrations of BHB were measured in whole blood, and plasma samples were analyzed for glucose, fatty acid (NEFA), insulin, glucagon, and electrolyte concentrations. The EDTA-anticoagulated blood samples were assessed for red blood cell indices, and smears were made for evaluation of red blood cell morphology. Outcomes were analyzed using repeated measures analysis. Overall least squares means (95% CI) of whole blood BHB concentrations between 1 h and d 11 relative to first treatment were 1.11 (0.95 to 1.30), 1.26 (1.07 to 1.47), 0.96 (0.81 to 1.13), and 1.53 (1.30 to 1.80) mmol/L for the GLU, PG, GLU+PG, and CTRL groups, respectively. Treatment with both glucose and propylene glycol led to a greater magnitude and more prolonged decrease in BHB concentrations compared with individual treatments. The NEFA and glucagon concentrations were lower immediately after treatment in GLU and GLU+PG groups compared with CTRL, and treatment with both glucose and propylene glycol was associated with a greater increase in glucose and insulin concentrations immediately after treatment compared with CTRL and GLU treatment alone. Treatments did not lead to differences in plasma mineral concentrations. We conclude that treatments varied in the magnitude of decreasing blood BHB concentrations in hyperketonemic postpartum cows, with the greatest decline after treatment with a combination of intravenous glucose and oral propylene glycol.     

In vitro and in vivo analysis of fatty acid effects on metabolism of 17β-estradiol and progesterone in dairy cows

Some studies have reported improved reproductive performance with dietary fat supplementation. This study examined effects of fatty acids with different lengths, or desaturation, or both, on metabolism of estradiol (E2) and progesterone (P4) in bovine liver slice incubations (experiments 1 and 2) and in vivo (experiment 3). In experiment 1, effects of fatty acids C16:0 (palmitic acid), C16:1 (palmitoleic acid), C18:1 (oleic acid), and C18:3 (linolenic acid) were evaluated at 30, 100, and 300 μM on P4 and E2 metabolism in vitro. In experiment 2, stearic acid (C18:0) and C18:3 were evaluated in the same incubation conditions. In experiment 1, all of the fatty acids had some significant inhibitory effect on metabolism of P4, E2, or both (300 μM C16:0 on E2; 100 μM C16:1 on E2; 300 μM C16:1 on both P4 and E2; 300 μM C18:1 on P4; and 100 and 300 μM C18:3 on both P4 and E2). In experiment 2, C18:3 (100 and 300 μM) but not C18:0 decreased P4 and E2 metabolism. Overall, the most profound increase (∼60%) in half-life of P4 and E2 was observed with incubations of 300 μM C18:3 in both in vitro experiments. Based on these in vitro results, in experiment 3 linseed oil (rich in C18:3) was supplemented into the abomasum and acute effects on metabolism of E2 and P4 were evaluated. Cows (n = 4) had endogenous E2 and P4 minimized (corpus luteum regressed, follicles aspirated) before receiving continuous intravenous infusion of E2 and P4 to analyze metabolic clearance rate for these hormones during abomasal infusion of saline (control) or 70 mL of linseed oil every 4 h for 28 h. Linseed oil infusion increased C18:3 in plasma by 46%; however, metabolic clearance rate for E2 and P4 were similar for control cows compared with linseed-treated cows. Thus, in vitro experiments indicated that E2 and P4 metabolism can be inhibited by high concentrations of C18:3. Nevertheless, in vivo, linseed oil did not acutely inhibit E2 and P4 metabolism, perhaps because insufficient C18:3 concentrations (increased to ∼8 μM) were achieved. Further research is needed to determine the mechanism(s) of fatty acid inhibition of P4 and E2 metabolism and to discover practical methods to mimic this effect in vivo.     

Effect of esterification with fatty acid of β-cryptoxanthin on its thermal stability and antioxidant activity by chemiluminescence method

β-Cryptoxanthin, a pro-vitamin A carotenoid of potential interest for health, was found in vegetables and fruits in both free and esterified forms. Stabilities of free β-cryptoxanthin (FCX) and its esters including β-cryptoxanthin laurate (CXL), β-cryptoxanthin myristate (CXM), and β-cryptoxanthin palmitate (CXP) against heat were studied by high performance liquid chromatography–mass spectrometry (HPLC–MS). All β-cryptoxanthin fatty acid esters were more stable against heat than FCX. The antioxidant activities of FCX and its esters on superoxide anion, hydroxide radical and singlet oxygen were investigated by the pyrogallol-luminol system, the CuSO₄–phenanthroline–Vc–H₂O₂ system, and OH–NaClO–H₂O₂ system, respectively, using chemiluminescence (CL) methods in vitro. The antioxidant activities of β-cryptoxanthin fatty acid esters were not significantly different in all methods, furthermore, no synergetic effects amongst them were obtained. These results suggest that esterification of OH groups with fatty acids might not affect antioxidant activity of β-cryptoxanthin but stabilize β-cryptoxanthin against heat degradation.     

Investigating the within-herd prevalence and risk factors for ketosis in dairy cattle in Ontario as diagnosed by the test-day concentration of β-hydroxybutyrate in milk

An observational study of 790 to over 3,000 herds was conducted to estimate the within-herd prevalence and cow-level risk factors for ketosis in dairy cattle in herds that participate in a Dairy Herd Improvement Association (DHIA) program. Ketosis or hyperketolactia (KET) was diagnosed as milk β-hydroxybutyrate ≥0.15 mmol/L at first DHIA test when tested within the first 30 d in milk. Seven hundred ninety-five herds providing at least 61 first milk tests from June 2014 to December 2015 were used to estimate the provincial within-herd prevalence of KET. All herds on DHIA in Ontario (n = 3,042) were used to construct cow-level multilevel logistic regression models to investigate the association of DHIA collected variables with the odds of KET at first DHIA milk test. Primiparous and multiparous animals were modeled independently. The cow-level KET prevalence in Ontario was 21%, with an average within-herd prevalence of 21% (standard deviation = 10.6) for dairy herds enrolled in a DHIA program. The prevalence of KET had a distinct seasonality with the lowest prevalence occurring from July to November. Automatic milking systems (AMS) were associated with increased within-herd prevalence, as well as increased odds of KET in multiparous animals at first test (odds ratio: 1.45; 95% confidence interval: 1.30 to 1.63). Jersey cattle had over 1.46 times higher odds of KET than Holstein cattle. Milk fat yield ≥1.12 kg/d at the last test of the previous lactation was associated with decreased odds of KET in the current lactation (odds ratio: 0.56; 95% confidence interval: 0.53 to 0.59). Increased days dry and longer calving intervals, for multiparous animals, and older age at first calving for primiparous animals increased the odds of KET at first test. This study confirms previous findings that increased days dry, longer calving intervals, and increased age at first calving are associated with increased odds of KET and is the first report of increased KET in herds with AMS and in relation to milk fat yield at the final test of the previous lactation. Feeding management on AMS herds likely contributes to the increased prevalence of KET and further work is required to investigate modifications to current management to minimize risk. Milk fat yield during the previous lactation may be representative of energy partitioning.     

Effect of dietary α-linolenic acid and vitamin E on the fatty acid composition, storage stability and sensory traits of rabbit meat

The synergistic effect of dietary linolenic acid and vitamin E on the oxidative stability and nutritional and eating characteristics of fresh and stored rabbit meat was studied. One-hundred hybrid male rabbits were divided into two homogenous groups and fed ad libitum two diets differing in the amount of sunflower and flaxseed and in the level of α-tocopherol, as follows: control diet: 0.08 kg kg(-1) sunflower, 50 mg kg(-1) α-tocopheryl-acetate and LNA-VE diet: 0.08 kg kg(-1) flaxseed, 200 mg kg(-1) α-tocopheryl-acetate. At 85 days, 20 rabbits per group were slaughtered and the thiobarbituric-acid reactive substances (TBA-RS), chemical composition, fatty acid profile and sensory quality were assessed on the longissimus dorsi muscles (fresh and stored for 8 days at 4?°C). The proximate composition of the fresh muscle was not significantly affected by the dietary treatment. Rabbits fed the LNA-VE diet showed a good capability to elongate and desaturate linolenic acid and this diet enriched the n-3 PUFA content of the meat without affecting its peroxidative stability. The sensory quality of the fresh and stored muscle was slightly affected by the dietary treatment, even though final tenderness (fresh meat) and overall acceptability (stored meat) of the LNA-VE rabbits showed significantly higher scores.     

Short communication: Effect of αS1-casein (CSN1S1) and κ-casein (CSN3) genotypes on milk composition in Murciano-Granadina goats

The effects of the caprine α_S1-casein (CSN1S1) polymorphisms on milk quality have been widely demonstrated. However, much less is known about the consequences of the κ-casein (CSN3) genotype on milk composition in goats. Moreover, the occurrence of interactions between CSN3 and CSN1S1 genotypes has not been investigated. In this study, an association analysis between CSN1S1 and CSN3 genotypes and milk quality traits was performed in 89 Murciano-Granadina goats. Total milk yield as well as total protein, fat, solids-not-fat, lactose, α_S1-casein (CSN1S1), and α_S2-casein (CSN1S2) contents were recorded every other month during a whole lactation (316 observations). Data analysis using a linear mixed model for repeated observations revealed no interaction between the CSN1S1 and CSN3 genotypes. With regard to the effect of the CSN3 locus, AB and BB genotypes were significantly associated with higher levels of total casein and protein content compared with the AA CSN3 genotype. In strong contrast with French breeds, the CSN1S1 genotype did not affect protein, casein, and fat concentrations in Murciano-Granadina goats. These results highlight the importance of taking into consideration the CSN3 genotype when performing selection for milk composition in dairy goats.     

Effect of ascorbic acid and protein concentration on the molecular weight profile of bovine serum albumin and β-lactoglobulin by γ-irradiation

The effects of ascorbic acid and protein concentration on the molecular weight size distribution of BSA and β-lactoglobulin were examined after irradiation of proteins at various doses. Gamma-irradiation of protein solutions caused disruption of the ordered structure of protein molecules resulting in degradation, cross-linking, and aggregation of the polypeptide chains. SDS–PAGE and gel permeation chromatography study showed that ascorbic acid protected the aggregation and degradation of proteins by scavenging oxygen radicals produced by irradiation and the effect of irradiation on protein conformation was more significant at lower concentrations of proteins.     

Effects of the peroxisome proliferator-activated receptor-α agonists clofibrate and fish oil on hepatic fatty acid metabolism in weaned dairy calves

Peroxisome proliferator-activated receptor-α (PPARα) agonists increase fatty acid oxidation in liver of nonruminants. If similar effects occur in dairy cattle, enhanced hepatic oxidative capacity could decrease circulating nonesterified fatty acids and hepatic triacylglycerol accumulation in periparturient cows. The objectives of this study were 1) to determine whether partitioning of fatty acid metabolism by liver slices from weaned Holstein calves treated with PPARα agonists in vivo is altered compared with partitioning by liver slices from control (untreated) calves, and 2) to measure in vitro metabolism of palmitate and oleate by bovine liver slices and relate these to mRNA abundance for key enzymes. Weaned male Holstein calves (7 wk old; n = 15) were assigned to 1 of 3 groups for a 5-d treatment period: control (untreated), clofibrate (62.5 mg/kg of BW), or fish oil (250 mg/kg of BW). Calves treated with clofibrate consumed less dry matter. Body weight, liver weight, liver weight:body weight ratio, blood nonesterified fatty acids, β-hydroxybutyrate, and liver composition were not significantly different among treatments. Liver slices were incubated for 2, 4, and 8 h to determine in vitro conversion of [1-¹⁴C] palmitate and [1-¹⁴C] oleate to CO₂, acid-soluble products, esterified products, and total metabolism. In liver slices incubated for 8 h, conversion of palmitate to CO₂ was greater for calves treated with clofibrate compared with control calves or calves treated with fish oil. Conversion of palmitate to esterified products, total palmitate metabolism, and metabolism of oleate were not different among treatments. Conversion of palmitate to CO₂ was greater than that from oleate for all treatments, but rates of total metabolism did not differ. Clofibrate increased or tended to increase liver expression of several PPARα target genes involved in fatty acid oxidation (e.g., ACADVL, ACOX1, CPT1A), whereas fish oil did not significantly affect genes associated with fatty acid oxidation but tended to increase DGAT1. Overall, our data indicated that bovine liver responded to clofibrate treatment but not fish oil, although increases in hepatic lipid metabolism were much less than those reported in rodents treated with clofibrate or fish oil. Applications of PPARα agonists may be of interest to increase the rate of hepatic fatty acid oxidation and decrease triacylglycerol accumulation in periparturient dairy cows.     

Short communication: Can the mammopathogenic Escherichia coli P4 strain have a direct role on the caseinolysis of milk observed during bovine mastitis?

During bacterial bovine mastitis, the quality of milk is altered because of caseinolysis. Endogenous potential actors in milk responsible for this caseinolysis have been well studied, unlike the exogenous bacterial ones. The aim of this study was to evaluate the direct role in caseinolysis of a mammopathogenic strain, Escherichia coli P4. Secretion of at least 4 extracellular bacterial caseinolytic enzymes was highlighted by zymography, in 3 different growth media, and at each bacterial growth state, suggesting that their expression was constitutive. Different experimental conditions to evaluate caseinolytic potential did not show any significant caseinolytic activity of E. coli P4 and of the 4 extracellular proteases detected, suggesting that the high caseinolysis observed during E. coli bovine mastitis does result from endogenous milk actors.     

Transfer rate of α-linolenic acid from abomasally infused flaxseed oil into milk fat and the effects on milk fatty acid composition in dairy cows

The objectives of the present study were to evaluate the transfer efficiency of α-linolenic acid (ALA) from the abomasum into milk fat, its interaction with milk fat content and yield, and the relationship between ALA and C16:0 in milk fat. Three rumen-fistulated multiparous Holstein cows at midlactation were used in a 3×3 Latin square design. Treatments consisted of abomasal infusion of (1) 110mL of water/d (control), (2) 110mL of flaxseed oil/d (low flaxseed oil, LFO), and (3) 220mL of flaxseed oil/d (high flaxseed oil, HFO). Experimental periods were continued for 2 wk and fat supplements were infused abomasally during the last 7 d of each period. Average dry matter intake and milk yield were not affected by oil infusion. Milk fat and lactose content tended to be greater with flaxseed infusion compared with the control. Plasma ALA was 2.9- and 4.0-fold greater with LFO and HFO, respectively. The apparent transfer efficiency of ALA to milk was 44.8 and 45.7% with LFO and HFO, respectively. The C16:0 content in milk fat was decreased by 3.59 and 5.25 percentage units, whereas the ALA content was increased by 1.68 and 3.09 percentage units with LFO and HFO, respectively. Similarly, C18:2n-6 was increased by 0.95 and 1.31 percentage units with LFA and HFO, respectively, without changes in other fatty acids (FA). Total polyunsaturated FA was 4.4 and 2.7% lower in the HFO and LFO, respectively, than in the control. Furthermore, C16:0 content in the milk fat was reduced to a greater extent than the increase in ALA content, as a 1.68 and 3.09 percentage unit increase occurred in ALA compared with a 3.6 and 5.25 percentage unit decrease in C16:0 for LFO and HFO, respectively, such that a negative correlation existed between ALA and C16:0 (r=-0.72). In conclusion, abomasal infusion of flaxseed oil dramatically increased the ALA content in plasma and milk fat. Because the replacement of C16:0 with ALA and C18:2n-6 occurred without changes in other FA presumed to be synthesized de novo in the mammary gland, this suggests that the preformed C16:0 was replaced, rather than being caused, by an overall suppression of de novo FA synthesis in the mammary gland.