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.

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 (57 d 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 28 d, 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 0.25 g of glucose/kg of body weight were performed 28 and 10d before expected parturition, as well as at 4 and 21 d 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.

The purpose of this experiment was to gain understanding on changes in energy partitioning when folic acid and vitamin B supplements, alone or combined, were given by weekly intramuscular injections from 3 wk before the expected calving date until 7 wk postpartum. Twenty-four multiparous cows were assigned to 6 blocks of 4 cows each according to previous 305-d lactation yield to either 0 or 320 mg of folic acid and 0 or 10 mg of vitamin B in a 2 × 2 factorial arrangement. Plasma concentration of folates was increased by folic acid supplement, and this increase was greater with the combined supplement. Vitamin B supplement increased plasma concentration of vitamin B. Even though postpartum energy balance was similar among treatments, postpartum body condition score was higher for cows receiving folic acid supplement compared with cows that did not. Milk yield of cows receiving folic acid supplement reached a plateau earlier than for cows that did not. Fat and protein, as well as total solid concentrations and yields, were unaffected by treatments. Postpartum plasma concentrations of glucose and insulin were higher and postpartum plasma concentration of nonesterified fatty acids was lower for cows that received weekly folic acid supplement compared with cows that did not. Plasma concentration of methylmalonic acid was low and unaffected by treatments, suggesting that vitamin B supply was not limiting, even for unsupplemented cows. Postpartum plasma concentrations of Cys, His, Phe, and Tyr were increased, whereas plasma concentration of Gly was decreased, by folic acid supplement. In the present study, supplementary folic acid altered energy partitioning in early lactation as suggested by similar milk total solid yield and postpartum energy balance, lower plasma nonesterified fatty acid concentration and body condition score losses, and higher plasma glucose and insulin concentrations for cows receiving folic acid supplement compared with cows that did not.

The present review focuses on methyl donor metabolism and nutrition in the periparturient and lactating dairy cow. Methyl donors are involved in one-carbon metabolism, which includes the folate and Met cycles. These cycles work in unison to support lipid, nucleotide, and protein synthesis, as well as methylation reactions and the maintenance of redox status. A key feature of one-carbon metabolism is the multi-step conversion of tetrahydrofolate to 5-methyltetrahyrofolate. Homocysteine and 5-methyltetrahyrofolate are utilized by vitamin B 12 -dependent Met synthase to couple the folate and Met cycles and generate Met. Methionine may also be remethylated from choline-derived betaine under the action of betaine hydroxymethyltransferase. Regardless, Met is converted within the Met cycle to S-adenosylmethionine, which is universally utilized in methyl-group transfer reactions including the synthesis of phosphatidylcholine. Homocysteine may also enter the transsulfuration pathway to generate glutathione or taurine for scavenging of reactive oxygen metabolites. In the transition cow, a high demand exists for compounds with a labile methyl group. Limited methyl group supply may contribute to inadequate hepatic phosphatidylcholine synthesis and hepatic triglyceride export, systemic oxidative stress, and compromised milk production. To minimize the perils associated with methyl donor deficiency, the peripartum cow relies on de novo methylneogenesis from tetrahydrofolate. In addition, dietary supplementation of rumen-protected folic acid, vitamin B 12 , Met, choline, and betaine are potential nutritional approaches to target one-carbon pools and improve methyl donor balance in transition cows. Such strategies have merit considering research demonstrating their ability to improve milk production efficiency, milk protein synthesis, hepatic health, and immune response. This review aims to summarize the current understanding of folic acid, vitamin B 12 , Met, choline, and betaine utilization in the dairy cow. Methyl donor co-supplementation, fatty acid feeding strategies that may optimize methyl donor supplementation efficacy, and potential epigenetic mechanisms are also considered.

Vitamin B 12 is synthesized by prokaryotes in the rumens of dairy cows-and this has implications in human nutrition since humans rely on consumption of dairy products for vitamin B 12 acquisition. However, the concentration of vitamin B 12 in milk is highly variable, and there is interest in determining what causes vitamin B 12 variability. We collected 92 temporally linked rumen, fecal, blood, and milk sample sets from Holstein cows at various stages of lactation fitted with rumen cannula and attempted to define which bacterial genera correlated well with vitamin B 12 abundance. The level of vitamin B 12 present in each sample was measured, and the bacterial population of each rumen, fecal, and milk sample (n ϭ 263) was analyzed by 16S rRNA-targeted amplicon sequencing of the V4 region. The bacterial populations present in the rumen, small intestine, and milk were highly dissimilar. Combined diet and lactation status had significant effects on the composition of the microbiota in the rumen and in the feces. A high ruminal concentration of vitamin B 12 was correlated with the increased abundance of Prevotella, while a low ruminal concentration of vitamin B 12 was correlated with increased abundance of Bacteroidetes, Ruminiclostridium, and Butyrivibrio. The ultimate concentration of vitamin B 12 is controlled by the complex interaction of several factors, including the composition of the microbiota. Bacterial consumption of vitamin B 12 in the rumen may be more important in determining overall levels than bacterial production. IMPORTANCE In this paper, we examined the microbiome of the bovine rumen, feces, and milk and attempted to understand how the bacterial communities at each site affected the production and movement of vitamin B 12 around the animal's body. It was determined that the composition of the bovine rumen microbiome correlates well with vitamin B 12 concentration, indicating that the rumen microbiota may be a good target for manipulation to improve production of this important vitamin.

Only bacteria can synthesize vitamin B12, and this requires adequate Co supply. The natural source of vitamin B12 in human diets comes from animal products, especially those from ruminants. This study aimed to describe variability regarding vitamin B12 concentration in milk among and within commercial dairy herds in early lactation. A secondary objective was to explore potential causes for this variability such as genetic variation and diet characteristics. In total, 399 dairy cows (135 primiparous and 264 multiparous; 386 Holstein and 13 Jersey cows) in 15 commercial herds were involved. Milk samples were taken at 27.4±4.1 and 55.4±4.1d in milk. Neither parity (primiparous vs. multiparous) nor sampling time affected milk concentrations of vitamin B12. Nevertheless, vitamin B12 concentration in milk was highly variable among and within dairy herds. The lowest vitamin B12 concentration in milk of cows was observed in the Jersey herd. Among herds, vitamin B12 concentration in milk ranged from 2,309 to 3,878 pg/mL; one glass (250mL) of milk from those herds would provide between 23 and 40% of the vitamin B12 recommended daily allowance. Among individual cows, however, this provision varied between 16 and 57% of the recommendation. In spite of the limited size of the studied population, the heritability value was 0.23, suggesting that genetic selection could modify milk vitamin B12 concentration. We observed a positive relationship between milk vitamin B12 concentration and dietary acid detergent fiber content and a negative relationship between milk concentration of vitamin B12 and dietary crude protein content.

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This study was undertaken to evaluate the effect of supplementation of folic acid and vitamin B on glucose and propionate metabolism. Twenty-four multiparous cows were assigned according to a complete block design in a 2 × 2 factorial arrangement to one of the following treatments: (1) saline 0.9% NaCl, (2) 320 mg of folic acid, (3) 10 mg of vitamin B, or (4) 320 mg of folic acid and 10 mg of vitamin B. Intramuscular injections were given weekly from 3 wk before the expected calving date until 9 wk postpartum. At 63 d in milk, d-[6,6-H]-glucose (16.5 mmol/h; jugular vein) and [1-C]-sodium propionate (13.9 mmol/h; ruminal vein) were simultaneously infused for 4 h; blood samples were collected from 2 to 4 h of the infusion period. Liver biopsies were carried out the following day. Supplements of folic acid and vitamin B respectively increased folate and vitamin B concentrations, both in milk and liver. Although dry matter intake was unaffected by treatments, milk and milk lactose yields tended to be lower by 5.0 and by 0.25 kg/d, respectively, for cows receiving the folic acid supplement. Plasma β-hydroxybutyrate concentration with the folic acid supplement followed the same tendency. Hepatic gene expression of methylmalonyl-CoA mutase and S-adenosylhomocysteine hydrolase was higher for cows receiving the combined folic acid and vitamin B supplement compared with cows receiving only the supplement of folic acid, whereas no treatment effect was noted for cows not receiving the folic acid supplement. Whole-body glucose rate of appearance and the proportion of whole-body glucose rate of appearance secreted in milk lactose decreased by 229 g/d and 5%, respectively, for animals receiving the folic acid supplement, concomitant with the lower milk lactose synthesis in these cows, indicating that supplementary folic acid may alter energy partitioning in cows. The absence of treatment effect on plasma concentrations of methylmalonic acid as well as on the proportion of glucose synthesized from propionate, averaging 60%, supports the fact that vitamin B supply was sufficient in control cows in the current study. Our results suggest that the folic acid supplement reduced glucose-derived lactose synthesis by redirecting glucose for other metabolic activity in the mammary gland or in other tissues.