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Abstract Background In the context of ecological studies, the Bayesian hierarchical Poisson model is of prime interest when studying the association between environmental exposure and rare diseases. However, adding spatially structured extra-variability in the model fitted to the data when such extra-variability does not exist conditionally on the covariates included in the model (over-fitting) may bias the estimation of the ecological association between covariates and relative risks toward the null. In order to investigate that possibility, a simulation study of the impact of introducing unnecessary residual spatial structure in the estimation model was conducted. Results In the case where no underlying extra-variability from the Poisson process exists, the simulation results show that models accounting for structured and unstructured residuals do not underestimate the ecological association, unless covariates have a very strong autocorrelation structure, i.e., 0.98 at 100 km on a territory of diameter 1000 km."

Most trace minerals (TM) are fed above dairy cow requirements in commercial herds but their fate and effects on dairy cows have not been well documented. In this study, we evaluated the effects of feeding short-term sulfate TM above recommendations on apparent total-tract digestibility of nutrients, rumen fermentation characteristics, serum concentrations, and milk yield and composition, as well as milk, fecal, and urinary TM excretion in midlactation dairy cows. Eight multiparous Holstein cows with an average body weight (± SD) of 684 ± 29 kg at 82 ± 10 DIM in a quadruple 2 × 2 crossover design were fed a basal diet, differing in sulfate TM supplement concentrations, to provide either 0.11, 17, and 63 (control; CON) or 0.95, 114, and 123 (high trace minerals; HTM) mg of dietary Co, Mn, and Zn per kilogram of DM, respectively. Each experimental period had a 21-d adaptation to the diet, followed by a 10-d sample collection period. Feed ingredients and total feces and urine were collected during 4 consecutive d and rumen fluid was collected 0, 1, 2, 4, and 6 h relative to feeding. Milk yield was recorded daily, and milk samples were collected on 4 consecutive milkings. Ingestion of Co, Mn, and Zn was higher for the HTM group compared with the CON group by 216%, 233%, and 93%, respectively. Dry matter intake averaged 25.0 (SE = 0.6) kg/d, and apparent total-tract digestibility of major nutrients was similar between treatments. High trace minerals had no measurable effect on ruminal pH, major volatile fatty acids, and protozoa counts. Isovalerate molar proportion was 9.4% greater for the HTM group compared with the CON group. Neither milk yield (43.5 kg/d; SE = 0.8) nor milk fat and protein concentrations differed between treatments. Milk urea nitrogen concentration was significantly higher for HTM (11.7 mg/dL) compared with CON (9.7 mg/dL; SE = 0.7). Fecal excretion of Co, Mn, and Zn increased by 223%, 198%, and 75%, respectively, for the HTM group compared with the CON group. Urinary excretions of TM were marginal compared with feces, and only urinary Co and Mn were significantly higher for HTM cows than CON cows, as was similarly obtained for serum Co and Mn concentrations. Milk TM yields were not modified by treatments. In summary, short-term dietary sulfate TM supply over the recommendation did not improve cow performance but significantly increased fecal TM excretion, which could affect TM accumulation in soils where manure is applied and could potentially result in leaching into nearby watersheds. Further studies are needed to evaluate the impact of high fecal TM excretion on the environment using the One Health approach. Moreover, the effects of TM oversupply on milk production and cow health should be evaluated by long-term experiments., (Copyright © 2024 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Synthesis of B vitamins by the rumen microbiota is usually sufficient to avoid the appearance of clinical deficiency symptoms in dairy cows under normal feeding conditions. Nevertheless, it is now generally accepted that vitamin deficiency is much more than the appearance of major functional and morphological symptoms. Subclinical deficiency, which is present as soon as the supply is lower than the need, causes cellular metabolic changes leading to a loss of metabolic efficiency. Folates and cobalamin, two B vitamins, share close metabolic relationships. Folates act as co-substrates in one-carbon metabolism, providing one-carbon unit for DNA synthesis and de novo synthesis of methyl groups for the methylation cycle. Cobalamin acts as a coenzyme for reactions in the metabolism of amino acids, odd-numbered chain fatty acids including propionate and de novo synthesis of methyl groups. Both vitamins are involved in reactions to support lipid and protein metabolism, nucleotide synthesis, methylation reactions and possibly, maintenance of redox status. Over the last decades, several studies have reported the beneficial effects of folic acid and vitamin B 12 supplements on lactation performance of dairy cows. These observations indicate that, even when cows are fed diets adequately balanced for energy and major nutrients, B-vitamin subclinical deficiency could be present. This condition reduces casein synthesis in the mammary gland and milk and milk component yields. Folic acid and vitamin B 12 supplements, especially when given together, may alter energy partitioning in dairy cows during early and mid-lactation as indicated by increased milk, energy-corrected milk, or milk component yields without affecting DM intake and BW or even with reductions in BW or body condition loss. Folate and cobalamin subclinical deficiency interferes with efficiency of gluconeogenesis and fatty acid oxidation and possibly alters responses to oxidative conditions. The present review aims to describe the metabolic pathways affected by folate and cobalamin supply and the consequences of a suboptimal supply on metabolic efficiency. The state of knowledge on the estimation of folate and cobalamin supply is also briefly mentioned., (Crown Copyright © 2023. Published by Elsevier B.V. All rights reserved.)

It is well established that the plasma metabolite profile changes during metabolic dysfunction, such as elevated non-esterified fatty acid (NEFA) release when body reserve mobilization is excessive in early lactation cows. Relationships between changes in plasma concentrations of metabolites caused by a metabolic impairment and the status of vitamins, such as folates and vitamin B 12 , have barely been studied in cattle. This study was undertaken to assess relationships between peripartum plasma concentrations of folates, vitamin B 12 , NEFA, and beta-hydroxybutyrate (BHB). Longitudinal data of 48 multiparous Holstein cows from 5 studies were taken from days -14 to 21 relative to calving. Blood samples were taken weekly before calving and either twice or thrice per week postpartum, and plasma was analyzed for folate, vitamin B 12 , NEFA, and BHB concentrations. Postpartum plasma NEFA and BHB concentrations were negatively related to plasma folate concentration at days -14 and -7 relative to parturition, whereas the opposite relationship was noted for the plasma vitamin B 12 :folate ratio. The plasma folate and NEFA areas under the curve from the whole studied period were negatively associated, and the opposite was observed with the association between the plasma vitamin B 12 :folate ratio and NEFA as well as the BHB areas under the curve. The results suggest that there is an increased use of folate for metabolic functions during elevated concentrations of plasma NEFA and BHB. Future research should focus on finding an optimal plasma vitamin B 12 :folate ratio to favor cow health during the challenging period of parturition.

The content, composition and variation of vitamin compounds in goat milk have been little studied. An experimental design was based on 28 commercial farms, selected considering the main feeding system (based on main forage and especially pasture access), goat breed (Alpine vs Saanen) and reproductive management (seasonal reproduction), in the main French goat milk production area. Each farm received two visits (spring and autumn) that included a survey on milk production conditions and bulk milk sampling. Milk vitamins (A, E, B 2 , B 6 , B 9 , B 12 ) and carotenoid concentrations plus colour indices were evaluated. A stepwise approach determined the variables of milk production conditions that significantly altered milk indicators. The main forage in the diet was the major factor altering goat milk vitamin and carotenoid concentrations and colour indices. Bulk milk from goats eating fresh grass as forage was richer in α-tocopherol (+64%), pyridoxal (+35%) and total vitamin B 6 (+31%), and b* index (characterising milk yellowness in the CIELAB colour space) was also higher (+12%) than in milk from goats eating conserved forages. In milk from goats eating fresh grass, concentrations of pyridoxamine, lutein and total carotenoids were higher than in milk of goats fed corn silage (+24, +118 and +101%, respectively), and retinol and α-tocopherol concentrations were higher than in milk of goats fed partially dehydrated grass (+45 and +55%). Vitamin B 2 concentration was higher in milk of goats eating fresh grass than in milk of goats fed hay or corn silage as forage (+10%). However, bulk milk when goats had access to fresh grass was significantly poorer in vitamin B 12 than when fed corn silage (-46%) and in γ-tocopherol (-31%) than when fed conserved forage. Alpine goats produced milk with higher vitamin B 2 and folate concentrations than Saanen goats (+18 and +14%, respectively). Additionally, the milk colour index that discriminates milks based on their yellow pigment contents was 7% higher in milk from Alpine than Saanen herds, but milk from Saanen goats was richer in lutein (+46%). Goat milks were richer in vitamins B 2 and B 12 and folates, but poorer in vitamin B 6 in autumn than in spring (+12, +133, +15 and -13%, respectively). This work highlights that goat milk vitamin and carotenoid concentrations and colour indices vary mainly according to the main forage of the diet and secondly according to the breed and season., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

As milk production has significantly increased over the past decade(s), existing estimates of the B-vitamin needs of the modern dairy cow are currently being reconsidered, as suboptimal B-vitamin supply may affect metabolic efficiency. At the same time, however, "true" (i.e., biologically active forms, excluding nonfunctional analogs) B-vitamin supply also cannot be adequately estimated by dietary intake, as the rumen microbiota has been shown to play a significant role in synthesis and utilization of B vitamins. Given their complex impact on the metabolism of dairy cows, incorporating these key nutrients into the next generation of mathematical models could help to better predict animal production and performance. Therefore, the purpose of this study was to generate hypotheses of regulation in the absence of supplemental B vitamins by creating empirical models, through a meta-analysis, to describe true B-vitamin supply to the cow (postruminal flow, PRF) and apparent ruminal synthesis (ARS). The database used for this meta-analysis consisted of 340 individual cow observations from 15 studies with 16 experiments, where diet and postruminal digesta samples were (post hoc) analyzed for content of B vitamins (B 1 , B 2 , B 3 , B 6 , B 9 , B 12 ). Equations of univariate and multivariate linear form were considered. Models describing ARS considered dry matter intake (DMI, kg/d), B-vitamin dietary concentration [mg/kg of dry matter (DM)] and rumen-level variables such as rumen digestible neutral detergent fiber (NDF) and starch (g/kg of DM), total volatile fatty acids (VFA, mM), acetate, propionate, butyrate, and valerate molar proportions (% of VFA), mean pH, and fractional rates of degradation of NDF and starch (%/h). Models describing PRF considered dietary-level driving variables such as DMI, B-vitamin dietary concentration (mg/kg of DM), starch and crude protein (g/kg of DM) and forage NDF (g/kg of DM). Equations developed were required to contain all significant slope parameters and contained no significant collinearity between driving variables. Concordance correlation coefficient was used to evaluate the models on the developmental data set due to data scarcity. Overall, modeling ARS yielded better-performing models compared with modeling PRF, and DMI was included in all prediction equations as a scalar variable. The B-vitamin dietary concentration had a negative effect on the ARS of B 1 , B 2 , B 3 , and B 6 but increased the PRF of B 2 and B 9 . The rumen digestible NDF concentration had a negative effect on the ARS of B 2 , B 3 , and B 6 , whereas rumen digestible starch concentration had a negative effect on the ARS of B 1 and a positive effect on the ARS of B 9 . In the best prediction models, the dietary starch increased PRF of B 1 , B 2 , and B 9 but decreased PRF of B 12 . The equations developed may be used to better understand the effect of diet and ruminal environment on the true supply of B vitamins to the dairy cow and stimulate the development of better-defined requirements in the future., (© 2022, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Biotin (B 8 ), folates (B 9 ), and vitamin B 12 (B 12 ) are involved and interrelated in several metabolic reactions related to energy and protein metabolism. We hypothesized that a low supply of one of the latter vitamins during the transition period would impair metabolic status. The purpose of this study was to evaluate the effect of B 8 supplementation on the response of lactation performance and selected energy and protein metabolites and hormones to a combined supplementation of B 9 and B 12 given to periparturient dairy cows, from d -21 to 21 relative to calving. A total of 32 multiparous Holstein cows housed in tie stalls were randomly assigned, according to their previous 305-d milk yield, to 8 incomplete blocks of 4 treatments: (1) a 2-mL weekly i.m. injection of saline (0.9% NaCl; B 8 -/B 9 B 12 -); (2) 20 mg/d of dietary B 8 (unprotected from ruminal degradation) and 2-mL weekly i.m. injection of 0.9% NaCl (B 8 +/B 9 B 12 -); (3) 2.6 g/d of dietary B 9 (unprotected) and 2-mL weekly i.m. injection of 10 mg of B 12 (B 8 -/B 9 B 12 +); and (4) 20 mg/d of dietary B 8 , 2.6 g/d of dietary B 9 , and weekly i.m. injection of 10 mg of B 12 (B 8 +/B 9 B 12 +) in a 2 × 2 factorial arrangement. Milk yield and dry matter intake were obtained daily and milk components weekly. Blood samples were taken weekly from d -21 to calving and 3 times per week from calving to 21 d following parturition. Prepartum plasma concentrations of glucose, insulin, nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHB), and adiponectin were unaffected by treatments. Biotin, B 9 , and B 12 supplements increased their respective concentrations in plasma and milk. Cows fed the B 8 supplement tended to have lower dry matter intake, but only cows in B 8 +/B 9 B 12 - had greater plasma concentrations of NEFA compared with B 8 -/B 9 B 12 -. Milk and total solid yields were greater by 13.5 and 13.9%, respectively, for B 8 -/B 9 B 12 + [45.5 (standard error, SE: 1.8) and 5.81 (0.22) kg/d, respectively] compared with B 8 -/B 9 B 12 - [40.1 (1.9) and 5.10 (0.23) kg/d, respectively], but these effects were suppressed when combined with the B 8 supplement. Cows in the B 8 -/B 9 B 12 + group had decreased plasma insulin and tended to have increased NEFA concentrations, but postpartum plasma concentrations of glucose, BHB, leptin, and adiponectin were not affected. These cows also mobilized more body fat reserves, as suggested by a tendency to increased plasma NEFA and more milk total solids compared with B 8 -/B 9 B 12 - cows. However, plasma concentrations of BHB and adiponectin were similar among treatments. This suggests that the B 9 and B 12 supplements enhanced efficiency of energy metabolism in early lactation cows. Folic acid and B 12 supplementation increased postpartum plasma Cys and homocysteine concentrations but did not affect plasma Met concentration, suggesting an upregulation of the transsulfuration pathway. In summary, our results showed that, under the current experimental conditions, increasing B 8 supply did not improve responses to the B 9 and B 12 supplementation., (Crown Copyright © 2022. Published by Elsevier B.V. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the Open Government Licence (https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/).)

Biotin (B 8 ), folate (B 9 ), and vitamin B 12 (B 12 ) are involved in several metabolic reactions related to energy metabolism. We hypothesized that a low supply of one of these vitamins during the transition period would impair metabolic status. This study was undertaken to assess the interaction between B 8 supplement and a supplementation of B 9 and B 12 regarding body weight (BW) change, dry matter intake, energy balance, and fatty acid (FA) compositions of colostrum and milk fat from d -21 to 21 relative to calving. Thirty-two multiparous Holstein cows housed in tie stalls were randomly assigned, according to their previous 305-d milk yield, to 8 incomplete blocks in 4 treatments: (1) a 2-mL weekly i.m. injection of saline (0.9% NaCl; B 8 -/B 9 B 12 -); (2) 20 mg/d of dietary B 8 (unprotected from ruminal degradation) and 2-mL weekly i.m. injection of 0.9% NaCl (B 8 +/B 9 B 12 -); (3) 2.6 g/d of dietary B 9 (unprotected) and 2-mL weekly i.m. injection of 10 mg of B 12 (B 8 -/B 9 B 12 +); (4) 20 mg/d of dietary B 8 , 2.6 g/d of dietary B 9 , and 2-mL weekly i.m. injection of 10 mg of B 12 (B 8 +/B 9 B 12 +) in a 2 × 2 factorial arrangement. Colostrum was sampled at first milking. and milk samples were collected weekly on 2 consecutive milkings and analyzed for FA composition. Body condition score and BW were recorded every week throughout the trial. Within the first 21 d of lactation, B 8 -/B 9 B 12 + cows had an increased milk yield by 13.5% [45.5 (standard error, SE: 1.8) kg/d] compared with B 8 -/B 9 B 12 - cows [40.1 (SE: 1.9)], whereas B 8 supplement had no effect. Even though body condition score was not affected by treatment, B 8 -/B 9 B 12 + cows had greater BW loss by 24 kg, suggesting higher mobilization of body reserves. Accordingly, milk de novo FA decreased and preformed FA concentration increased in B 8 -/B 9 B 12 + cows compared with B 8 -/B 9 B 12 - cows. In addition, cows in the B 8 +/B 9 B 12 - group had decreased milk de novo FA and increased preformed FA concentration compared with B 8 -/B 9 B 12 - cows. Treatment had no effect on colostrum preformed FA concentration. Supplemental B 8 decreased concentrations of ruminal biohydrogenation intermediates and odd- and branched-chain FA in colostrum and milk fat. Moreover, postpartum dry matter intake for B 8 + cows tended to be lower by 1.6 kg/d. These results could indicate ruminal perturbation caused by the B 8 supplement, which was not protected from rumen degradation. Under the conditions of the current study, in contrast to B 8 +/B 9 B 12 - cows, B 8 -/B 9 B 12 + cows produced more milk without increasing dry matter intake, although these cows had greater body fat mobilization in early lactation as suggested by the FA profile and BW loss., (Crown Copyright © 2022. Published by Elsevier B.V. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the Open Government Licence (https://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/).)

The transition from a liquid- to a solid-based diet involves several adaptations in calves. Development of ruminal function is likely to alter B vitamin and choline supply, although little is known about the extent of these changes relative to the calf's requirements and consequences for the calf around weaning. Moreover, literature data are equivocal concerning the need to supplement B vitamins and choline through weaning and transition phase of the dairy calf. To evaluate the effect of increasing B vitamin and choline supply on performance, 61 Holstein calves were individually housed and raised from birth to 13 wk of age. Calves were fed milk replacer (28% crude protein, 15% fat) up to 1.6 kg of dry matter (DM)/d at 15% solids (3 times/d) from birth to 4 wk of age. At that time, calves were randomly assigned to one of 3 treatments: a rumen-protected blend of B vitamins and choline (RPBV); a 30:70 mix of a nonprotected blend of B vitamins and choline and fat (UPBV); or fat only, used as control (CTRL). Calves were maintained on milk replacer and offered ad libitum quantities of a starter grain (25.5% crude protein) specifically formulated to supply all essential amino acids with no added B vitamins or choline. The supplements were provided in gel capsules and administered once a day to each calf in quantities corresponding to 0.39 and 0.28% of the previous day's starter DM intake for the vitamin blends and control, respectively. Calves were weaned gradually from d 49 to 63. Body weight and stature were measured, and blood was collected and analyzed for hematocrit, plasma urea nitrogen, β-hydroxybutyrate, folates, and vitamin B 12 . Body weight and stature were similar among treatments. Overall gain (0.99 kg/d), DM intake (1.90 kg of DM/d), and feed efficiency (0.52) were not affected by vitamin supplementation. Plasma vitamin B 12 concentrations were not different between RPBV and UPBV but tended to be higher at the end of weaning and were greater postweaning in RPBV and UPBV treatments compared with CTRL. Both forms of the vitamin blend effectively improved vitamin B 12 status postweaning with no effect on folate status. No differences were observed in other blood measurements. Under conditions of this study, additional B vitamins and choline did not improve calf performance before, during, or after weaning., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Clinical symptoms of B vitamin deficiency are rarely observed in ruminants because these vitamins are synthesized by the rumen microbiota. However, over the last decades, numerous reports of beneficial effects on production and metabolic efficiency of dairy cows have been published supporting that, under some conditions, B vitamin subclinical deficiency is present in these animals. Due to their roles as coenzymes or cofactors in major metabolic pathways, an adequate supply in B vitamins is critical to optimize metabolic efficiency. Nowadays, taking into account the growing interest for the Smart Farming concept, fulfilling ruminant requirements for B vitamins according to their physiological stage under different feeding management cannot be neglected. In dairy cows, B vitamin supply is greatly dependent of the activity of the ruminal microbiota. Indeed, the amount of vitamins reaching the small intestine is dependent of the utilization of the vitamins provided by the diet and their synthesis by the microorganisms present in the rumen. The two major challenges faced to determine B vitamin status of ruminants are the difficulty to estimate B vitamin supply due to the lack of knowledge on factors driving the fate of B vitamins in the digestive tract, especially in the rumen, and the choice and thresholds of biomarkers reflecting adequately the animal status. The present paper aims to present the actual state of knowledge on the methodological approaches used to estimate B vitamin supply and status of ruminant and to point out future research orientations., (Crown Copyright © 2020. Published by Elsevier Inc. All rights reserved.)

The aim of the study was to evaluate glucose and insulin metabolism of cows receiving a supplementation of biotin (B 8 ), folic acid (B 9 ), and vitamin B 12 (B 12 ) during the transition period. According to a 2 × 2 factorial arrangement, 32 cows were randomly assigned to 9 incomplete blocks according to their previous 305-d milk yield. Within each block, cows were randomly assigned to 1 of the following levels of biotin from -27 to 28 d relative to the parturition: (1) no biotin supplement (B 8 -) or (2) 20 mg/d of dietary biotin (B 8 +). Within each level of biotin, the cows received either (1) 2-mL weekly intramuscular injections of saline 0.9% NaCl (B 9 B 12 -) or (2) 2.6 g/d of dietary folic acid and 2-mL weekly intramuscular injections of 10 mg of vitamin B 12 (B 9 B 12 +). An intravenous glucose tolerance test was performed at 25 d in milk. Baseline plasma glucagon, glucose, and nonesterified fatty acid concentrations did not differ among treatments. For B 9 B 12 + cows, baseline plasma insulin concentration and maximal glucose concentration after glucose administration were greater when also combined with biotin compared with no biotin combination, whereas there was no effect in B 9 B 12 - cows. There was no treatment effect on time to reach half-maximal glucose and insulin concentrations, glucose positive incremental area under the curve, and glucose and insulin clearance rates. Regarding insulin results, maximal plasma concentration and positive incremental area under the curve were respectively 51 and 74% greater for cows receiving the B 8 supplement than for cows who did not. Moreover, plasma nonesterified fatty acid concentration nadir tended to be reached later for B 8 cows. Insulin peak was reached earlier for cows in the group B 9 B 12 + than cows in B 9 B 12 -, regardless of B 8 supplementation. Under the current conditions, our results suggested that cows receiving a B 8 supplement had a reduced insulin sensitivity in early lactation. Insulin response was faster for B 9 B 12 + cows, but this was not translated into further improvements following the glucose administration challenge., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Many studies have shown that metabolic efficiency of ruminants can be significantly decreased when B-vitamin supply is insufficient. Under the present state of knowledge, the amounts of B vitamins available for intestinal absorption cannot be predicted based on diet composition. Therefore, in an attempt to increase our understanding of the effects of dietary factors, on B-vitamin supply for dairy cows, the effects of increasing amounts of extruded linseed in diets based on hay (permanent grassland hay, H; Experiment 1) or corn silage (CS; Experiment 2) on apparent ruminal synthesis (ARS) of thiamin, riboflavin, niacin, vitamin B6, folates and vitamin B12 were evaluated. In each experiment, four lactating Holstein cows fitted with cannulas in the rumen and the proximal duodenum were used in a 4 × 4 Latin square design. In both experiments, the dietary treatments consisted of an increasing supply of extruded linseed representing 0%, 5%, 10% or 15% of diet DM. The forage : concentrate ratios were 50 : 50 and 60 : 40 for Experiments 1 and 2, respectively. Duodenal flow was determined using YbCl3 as a marker. The ARS of each B vitamin was calculated as duodenal flow - daily intake. In both experiments, treatments did not affect thiamin, riboflavin, niacin and vitamin B12 duodenal flow or ARS. Increasing the dietary concentration of extruded linseed decreased folate intake in Experiment 1 and vitamin B6 intake in Experiment 2 but resulted in a greater duodenal flow of vitamin B6 and folates regardless of the forage used in basal diet. Greater dietary linseed concentrations decreased vitamin B6 apparent degradation in the rumen in CS-based diet only and increased folate ARS in both H- and CS-based diets. Increasing linseed concentration of isonitrogenous and isoenergetic diets increased vitamin B6 and folate supply to dairy cows, both with H- and CS-based diets.

The purpose of this study was (1) to predict the quantitative concentration of vitamin B 12 in milk using mid-infrared (MIR) spectrometry, and (2) to evaluate the potential of MIR spectra to discriminate different clusters of records based on their B 12 concentration. Milk samples were collected from 4,340 Holstein cows between 3 and 592 d in milk and located in 100 herds. Samples were taken using in-line milk meters and divided into 2 aliquots: one for MIR spectrometry and the other for B 12 concentration reference analyses by radioassay. Analyses were performed on 311 selected spectral wavelengths. A partial least squares regression model was built to quantify B 12 concentration. Discriminant analysis was used to isolate B 12 concentration clusters. A B 12 concentration threshold was set at 442 ng/dL, because this represents the cutoff value for a 250-mL glass of milk to fulfill 46% of the daily vitamin B 12 recommended dietary allowance for individuals 14 yr or older. For each analysis, records coming from two-thirds of herds were used to calibrate prediction equations, and the remaining records (one-third of herds for validation) were used to assess the prediction performance. In the case of discriminant analysis, validation sets were divided into evaluation sets (one-third of herds) to obtain alternate probability cutoffs and in test sets (two-thirds of herds) to validate equations. Spectral and B 12 concentration outliers were identified by calculating standardized Mahalanobis distance and with a residual analysis, respectively (n = 3,154). Regarding quantitative B 12 concentration, cross-validation and validation coefficients of determination averaged 0.51 and 0.46, respectively, which are relatively low, which would limit the potential use of the developed quantitative equations. In addition, root mean square errors of prediction of cross validation and validation sets averaged 88.9 and 94.7 ng/dL, respectively. Area under the receiver operating characteristic curve of test sets averaged 0.81 based on the 442 ng/dL threshold, which could be considered to represent good accuracy of classification. However, the false discovery rate averaged 36%. In summary, models predicting quantitative B 12 concentration had low cross-validation and validation coefficients of determination, limiting their use, but the proposed discriminant models could be used to identify milk samples with naturally high B12., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this work is to compare the ability of three spectroscopy techniques: molecular fluorescence, near-infrared (NIR), and mid-infrared with attenuated total reflectance (MIR-ATR) spectroscopy to predict the concentrations of 8 carotenoids, 6 vitamins and 22 fatty acids (FA) in cow's milk. A dataset was built through the analysis of 242 frozen milk samples from different experiments. The milk compounds were analysed using reference methods and by NIR, MIR-ATR, and fluorescence to establish different predictive models. NIR spectroscopy allowed for better prediction of cis9-β-carotene, β-cryptoxanthin and the sum of carotenoids than the other techniques, with a coefficient of cross-validation in calibration (R 2 CV) > 0.60 and a coefficient of determination in validation (R 2 V) > 0.50. Their standard errors of prediction (SEP) were equal to 0.01, except for the sum of carotenoids (SEP = 0.15). However, MIR-ATR and fluorescence seem usable for the prediction of lutein and all-trans-β-carotene, respectively. These three spectroscopy methods did not allow us to predict (R 2 CV < 0.30) vitamin contents except, for vitamin A (the best R²CV = 0.65 with NIR and SEP = 0.15) and α-tocopherol (the best R²CV = 0.56 with MIR-ATR and SEP = 0.41), but all R²V were <0.30. NIR spectroscopy yielded the best prediction of the selected milk FA.

Glycogen, a complex polysaccharide, is the form of storage of glucose in mammals that can be released rapidly when needed. Recent studies have mainly reported hepatic glycogen concentration for early-lactating cows, when the energy demand is higher than the energy supply from dry matter intake, driving the cow to use the energy stored as hepatic glycogen. Generally, liver samples are obtained through percutaneous needle biopsies in the right lobe of the liver. Our objective was to analyze the variation of glycogen concentration in the livers of Holstein and Jersey cows among different liver locations representing all lobes, to evaluate whether samples obtained by liver biopsies are representative of the whole organ. Liver from 10 culled lactating cows (5 Holstein and 5 Jersey cows) from 30 to 113 mo of age at slaughter were obtained. Each liver was sampled no more than 3 h after death on the following sites: 3 sites in the right lobe (1 to 3), 2 in the diaphragmatic surface of the left lobe (4 and 5), 3 in the visceral surface of the left lobe (6 to 8), 1 in the quadrate lobe (9), and 1 in the caudate lobe (10). Samples were snap frozen in liquid N 2 and were then analyzed for glucose concentration after conversion of glycogen to glucose using amyloglucosidase (EC 3.2.1.3). Glycogen results are reported as grams of glucose per 100 g of wet weight of liver (i.e., percent of wet weight of liver). Liver weights averaged 5.1 [standard deviation (SD) 1.2, minimum 3.3, maximum 6.2] kg for Holstein and 6.0 (SD 1.8, minimum 4.7, maximum 8.9) kg for Jersey cows. Holstein cows [1.31, standard error of the mean (SEM) 0.05% of wet weight] had greater liver glycogen concentration than did Jersey cows (0.75, SEM 0.05% of wet weight). No significant difference was noted among the 10 liver locations regarding glycogen concentration and averaged, for both breeds, 1.03% of wet weight (SEM 0.10). These results suggest that, in dairy cows, percutaneous needle liver biopsy in the right lobe is an accurate technique to fairly extrapolate glycogen concentration of the whole organ., (Crown Copyright © 2020. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

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  = 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., (© Crown copyright 2020.)

The objective of this cross-sectional study was to assess the variability of plasma folate and vitamin B 12 concentrations in lactating Holstein cows across the United States and Canada. We also evaluated the effect of diet composition and cow characteristics on folate and vitamin B 12 plasma vitamin concentrations. A total of 22 and 24 US and Canadian dairy herds were enrolled, totaling 427 and 476 cows at 10 to 197 days in milk across all US and Canadian herds, respectively. Blood samples were taken to analyze plasma folate and vitamin B 12 concentrations, and ingredients of the diet were collected to determine nutrient composition. To reduce the number of interdependent variables in the analysis of the association of diet composition with plasma vitamin concentrations, we conducted a principal component analysis. Plasma folate concentrations were lower for US cows [13.4 ng/mL, 95% confidence interval (CI): 12.7-14.2] than for Canadian cows (14.5 ng/mL, 95% CI: 13.7-15.2), and the opposite was observed for plasma vitamin B 12 concentrations (US 206 pg/mL, 95% CI: 192-221; Canada 170 pg/mL, 95% CI: 159-181). The highest plasma concentrations of both vitamins were observed in the Northwest region of the United States (Oregon and Washington). Cows in California had the lowest plasma folate concentrations, and cows in Québec and New York State had the lowest plasma vitamin B 12 concentrations. Plasma folate concentrations were higher for multiparous than for primiparous cows and plasma vitamin B 12 concentrations progressively increased from parity 1 to 3 and higher. For both studied vitamins, plasma concentrations were lower at 0 to 55 than at 56 to 200 days in milk. Of 3 principal components, the one associated with dietary carbohydrates was significantly correlated with plasma folate and vitamin B 12 concentrations. Indeed, plasma folate concentrations decreased with dietary fiber concentrations (i.e., neutral and acid detergent fibers and lignin) and increased with dietary nonfiber carbohydrate concentrations. We obtained the opposite results for plasma vitamin B 12 concentrations. Both multivariable models explained 41% (pseudo-R 2 ) of the variation in plasma folate and vitamin B 12 concentrations. Information gathered in this study is the first step toward determining sources of variation in plasma folate and vitamin B 12 concentrations, as well as the vitamin status of cows., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Cows undergoing a negative energy balance (NEB) often experience a state of immunosuppression and are at greater risk of infectious diseases. The present study aimed to evaluate the impact of a folic acid and vitamin B12 supplement and feed restriction on several immune parameters. Sixteen cows at 45 ± 3 days in milk were assigned to 8 blocks of 2 cows each according to each cow's milk production in the previous week, and within each block, the cows randomly received weekly intramuscular injections of either saline or 320 mg of folic acid and 10 mg of vitamin B12 for 5 weeks. During week 5, the cows were fed 75% of their ad libitum intake for 4 days. Blood samples were taken before the beginning of the experiment, just before feed restriction and after 3 days of feed restriction, in order to evaluate blood cell populations, the phagocytosis capacity and oxidative burst of polymorphonuclear leukocytes (PMNs), the proliferation of peripheral blood mononuclear cells (PBMCs) and concentrations of non-esterified fatty acids (NEFAs) and β-hydroxybutyrate. The vitamin supplement did not affect any of the tested variables except milk fat and lactose content. Feed restriction reduced milk production and increased the concentration of NEFAs. Feed restriction did not affect blood cell populations but did reduce the percentage of PMN positive for oxidative burst after stimulation with phorbol 12-myristate 13-acetate. The proliferation of PBMCs was reduced when the cell culture medium was supplemented with sera collected during the feed restriction. In conclusion, feed restriction affected the functions of PMN and PBMC and this effect was not prevented by the folic acid and vitamin B12 supplement. These results support the hypothesis that the greater risk of infectious diseases in cows experiencing a NEB is related to impaired immune cell functions by high circulating concentration of NEFAs.

As vitamin B12 is only synthesized by bacteria, ruminant products, especially dairy products, are excellent sources of this vitamin. This study aims to identify if diet and cow characteristics could affect vitamin B12 concentration in milk of dairy cows. Information on 1484 first, 1093 second and 1763 third and greater parity Holstein cows in 100 herds was collected during three consecutive milkings. During the first morning milking, all dietary ingredients given to cows were sampled and quantities offered were recorded throughout the day. Nutrient composition of ingredients was obtained by wet chemistry to reconstitute nutrient composition of the ration. Milk samples were taken with in-line milk meters during the evening milking of the 1st day and the morning milking of the 2nd day and were analyzed for vitamin B12 concentration. Milk yields were recorded and milk components were separately analyzed for each milking. Daily vitamin B12 concentration in milk was obtained using morning and evening vitamin B12 concentrations weighted with respective milk yield, then divided by daily yield. To decrease the number of interdependent variables to include in the multivariable model, a principal component analysis was carried out. Daily milk concentration of vitamin B12 averaged 3809±80 pg/ml, 4178±79 pg/ml and 4399±77 pg/ml for first, second and third, and greater lactation cows. Out of 11 principal components, six were significantly related to daily milk concentration of vitamin B12 when entered in the multivariable model. Results suggested that vitamin B12 concentration in milk was positively related to percentage of fiber and negatively related to starch as well as energy of the diet. Negative relationships were noted between vitamin B12 concentration in milk and milk yield as well as milk lactose concentration and positive relationships were observed between vitamin B12 concentration in milk and milk fat as well as protein concentrations. The percentages of chopped mixed silage and commercial energy supplement in the diet as well as cow BW were positively related to vitamin B12 in milk and percentages of baled mixed silage, corn and commercial protein supplement in the ration were negatively related to vitamin B12 concentration in milk. The pseudo-R2 of the model was low (52%) suggesting that diet and cow characteristics have moderate impact on vitamin B12 concentration in milk. Moreover, when entering solely the principal component related to milk production in the model, the pseudo-R2 was 46%. In conclusion, it suggests that studied diet characteristics have a marginal impact on vitamin B12 concentration in milk variation.

The present experiment was conducted to determine whether, during periods of negative energy balance, the increase in glucose availability, despite similar DMI and greater milk production, induced by a combined supplement of folic acid and vitamin B 12 was related to effects of insulin on metabolism. Sixteen multiparous Holstein cows averaging 45 days in milk (standard deviation: 3) were assigned to 8 blocks of 2 animals each according to their milk production (45 kg/d; standard deviation: 6) during the week preceding the beginning of the experiment. Within each block, they received weekly intramuscular injections of either saline (CON) or folic acid and vitamin B 12 (VIT) during 5 consecutive weeks. During the last week, the cows were fed 75% of their ad libitum intake during 4 d. Blood samples were taken the morning before starting the feed restriction and on the third day of feed restriction. On the fourth day of feed restriction, the daily meal was not served and an intravenous glucose tolerance test was performed. During the 4 wk preceding the feed restriction, milk production and DMI were not affected by treatments. During the feed restriction, the vitamin supplement tended to decrease milk fat concentration and increase milk concentration of lactose. Plasma concentrations of homocysteine, Ile, Leu, Val, and branched-chain AA increased in VIT cows during the restriction but not in CON cows. During the glucose tolerance test, insulin peak height was lower and insulin incremental positive area under the curve tended to be lower for VIT than for CON [83 (95% confidence interval, CI: 64-108) vs. 123 (95% CI: 84-180) µg·180 min/L, respectively]. Free fatty acid nadir was reached earlier for VIT than for CON [34 (95% CI: 26-43) vs. 46 (95% CI: 31-57) min, respectively]. Glucose area under the curve, clearance rate and peak height, insulin time to reach the peak and clearance rate, and free fatty acid nadir did not differ between VIT and CON. The reduction in insulin release during a glucose tolerance test without changes in glucose clearance rate or area under the curve suggests that the vitamin supplement improved insulin sensitivity in feed-restricted lactating dairy cows., (Crown Copyright © 2019. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

Methylmalonic acidemia (MMA), an organic acidemia characterized by metabolic instability and multiorgan complications, is most frequently caused by mutations in methylmalonyl-CoA mutase (MUT). To define the metabolic adaptations in MMA in acute and chronic settings, we studied a mouse model generated by transgenic expression of Mut in the muscle. Mut-/-;TgINS-MCK-Mut mice accurately replicate the hepatorenal mitochondriopathy and growth failure seen in severely affected patients and were used to characterize the response to fasting. The hepatic transcriptome in MMA mice was characterized by the chronic activation of stress-related pathways and an aberrant fasting response when compared with controls. A key metabolic regulator, Fgf21, emerged as a significantly dysregulated transcript in mice and was subsequently studied in a large patient cohort. The concentration of plasma FGF21 in MMA patients correlated with disease subtype, growth indices, and markers of mitochondrial dysfunction but was not affected by renal disease. Restoration of liver Mut activity, by transgenesis and liver-directed gene therapy in mice or liver transplantation in patients, drastically reduced plasma FGF21 and was associated with improved outcomes. Our studies identify mitocellular hormesis as a hepatic adaptation to metabolic stress in MMA and define FGF21 as a highly predictive disease biomarker.

The objective of the experiment was to determine if factors such as endosperm type (floury vs. vitreous) and particle size (fine vs. medium) of dry corn grain, known to affect starch digestibility in the rumen, modify apparent ruminal synthesis and duodenal flow of B vitamins in lactating dairy cows. Eight lactating multiparous Holstein cows equipped with rumen and duodenal cannulas were assigned randomly to a treatment sequence according to a 2 × 2 factorial arrangement in duplicate 4 × 4 Latin square design experiment. Duration of each experimental period was 21 d. When expressed per unit of dry matter intake (DMI), floury treatments increased duodenal flow and apparent ruminal synthesis of niacin and folates but tended to increase apparent degradation of thiamin in the rumen. Duodenal flow of thiamin, riboflavin, niacin, folates, and vitamin B 12 , expressed per unit of DMI, decreased with an increase in particle size. Similarly, apparent degradation of thiamin and riboflavin was greater and apparent synthesis of niacin, folates, and vitamin B 12 was reduced when cows were fed coarser dry corn grain particles. Neither endosperm type nor particle size had an effect on duodenal flow and apparent ruminal synthesis of vitamin B 6 . Apparent ruminal syntheses, expressed per unit of DMI, of all studied B vitamins but thiamin were negatively correlated with apparent ruminal digestibility of neutral detergent fiber. Duodenal flow of microbial N was positively correlated with apparent ruminal synthesis of riboflavin, niacin, vitamin B 6 , and folates. Under the conditions of the present experiment, except for thiamin, the effects of factors increasing starch digestibility of dry corn grain in the rumen on the amounts of B vitamins available for absorption by the dairy cow seem to be mediated through differences on ruminal digestibility of neutral detergent fiber and, to a lesser extent, on duodenal microbial N flow., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study was undertaken to evaluate the relationship between plasma and milk concentrations of vitamin B 12 as well as the relationship between plasma or milk concentrations of vitamin B 12 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 B 12 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 B 12 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 B 12 of first and second parities was lower than plasma concentration of third and more lactation cows. Milk vitamin B 12 concentration was positively correlated with plasma vitamin B 12 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 B 12 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 B 12 concentrations than cows with normal plasma FFA concentration. No relationship between vitamin B 12 concentration in milk or plasma and blood BHB concentration and hyperketonemia was noted. In summary, milk is not well correlated with plasma vitamin B 12 concentration for HO. It could be hypothesized that elevated plasma concentration of FFA could have a negative effect on the use of vitamin B 12 by cow cells, which increases the concentration of the vitamin in plasma and its secretion in milk., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The present study compares the bioavailability of vitamin B 12 (B 12 ) of dairy products or synthetic B 12 , using the pig as an experimental model for humans. Eleven pigs were used in a cross-over design to assess the net portal drained viscera (PDV) flux of blood plasma B 12 after ingestion of tofu (TF; devoid of B 12 ), Swiss cheese (SC), Cheddar cheese (CC), yogurt (YG), and synthetic B 12 (TB 12 ; TF supplemented with cyanocobalamin), providing a total of 25 µg of B 12 each. PDV blood plasma flow for SC and CC were higher than for TF and TB 12 ( p ≤ 0.04) whereas YG was higher than TF ( p = 0.05). Porto-arterial difference of blood plasma B 12 concentrations were higher for CC and TB 12 than for TF and YG ( p ≤ 0.04) but not different from SC ( p ≥ 0.15). Net PDV flux of B 12 was only different from zero for CC. However, the net PDV flux of B 12 for CC was not different from SC or TB 12 . Cumulative net PDV flux of B 12 for SC, TB 12 , and CC were 2.9, 4.4, and 8.3 µg 23 h post-meal, corresponding to a bioavailability of 11.6%, 17.5%, and 33.0%, respectively. In conclusion, CC had the best bioavailability of B 12 among the tested dairy products or compared to synthetic B 12 .

In pig, the assessment of bioavailability of dietary trace minerals with classical approaches such as relative bioavailability estimates or digestive tract balances have often generated inconsistent responses. In the present study, net portal-drained-viscera fluxes were monitored after a meal to assess intestinal absorption of zinc (Zn) or copper (Cu) according to dietary sources and levels of these trace minerals. Twelve pigs were surgically equipped with portal and carotid catheters and a portal ultrasonic flow probe for 12-h postprandial measurements. In a cross-over design, pigs received boluses of inorganic (I) or organic (O) dietary Cu and Zn at adequate (A, 20 and 200mg, respectively) or high (H, 40 and 400mg, respectively) level just before a 0.8-kg meal (semi-purified diet). Whatever treatments, arterial Zn increased by 72% at 45min postprandial and gradually declined thereafter (P<0.01). Arterial Zn were greater by 11% after O than I (P=0.02) and by 19% after H than A (P<0.01) meals. Net portal-drained-viscera fluxes of Zn during the first 240min postprandial were greater by 44% after O than I (P=0.10) and by 51% after H than A (P=0.07) meals. For Cu, portal-drained-viscera fluxes of Cu up to 240min postprandial were greater (P=0.03) after A than H meals. Those results suggest that Zn is absorbed rapidly, likely in the upper digestive tract of pigs and, whatever dietary levels, more efficiently after O meals. It appears that H levels of both Zn and Cu interfered with intestinal absorption of Cu and/or stimulate post-absorption enterocyte sequestration of this mineral., (Crown Copyright © 2017. Published by Elsevier GmbH. All rights reserved.)

The effect of fat supplements (FS) providing different proportions of saturated (SFA) and unsaturated (UFA) fatty acids on supply, apparent ruminal synthesis (ARS), and duodenal flow (DF) of some B vitamins (thiamine, riboflavin, niacin, vitamin B 6 , folates, and vitamin B 12 ) were evaluated in an experiment using 8 ruminally and duodenally cannulated lactating Holstein cows. The experiment was a replicated 4 × 4 Latin square design with 21-d treatment periods. The 4 treatments were a control diet without fatty acid supplement and 3 diets with 2.5% additional fatty acids from supplements containing (1) SFA, (2) an intermediate mixture of SFA and UFA, or (3) UFA. All diets were served as a total mixed ration once daily at 115% of the expected intake. B-vitamin concentrations were analyzed in feed and duodenal digesta. Apparent ruminal synthesis of each B vitamin was calculated as the DF minus the intake. B-vitamin concentrations were similar among the 4 treatments; consequently, daily intake of the vitamins followed the same pattern as dry matter intake. Adding FS decreased B-vitamin intakes (except vitamin B 12 ), as did increasing the proportion of UFA. Riboflavin and niacin DF and ARS, expressed as total daily amount or per unit of dry matter intake, were not affected by FS, but increasing the proportion of UFA decreased riboflavin and niacin DF and ARS. Fat supplements decreased DF of vitamin B 6 , expressed either as total daily amount or per unit of dry matter intake. No treatment effects were observed on total daily folate DF and ARS. However, when expressed per unit of dry matter intake, folate DF and ARS were greater when cows were fed FS and they increased linearly with the proportion of UFA in the supplement. Inclusion of fat supplements into the dairy cow diet had a limited effect on the fate of most B vitamins in the rumen although increasing the proportion of UFA in the FS linearly decreased apparent synthesis of riboflavin and niacin in the rumen and the amounts of these vitamins reaching the small intestine., (Crown Copyright © 2017 Published by FASS and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

Apparent ruminal synthesis and duodenal flow of thiamin, riboflavin, niacin, vitamin B 6 , folates, and vitamin B 12 were evaluated in an experiment using 15 ruminally and duodenally cannulated lactating Holstein cows fed a basal diet, according to a crossover design, supplemented or not with 56 g/d of Saccharomyces cerevisiae fermentation product. Duration of the treatment period was 28 d. The basal ration had 28% neutral detergent fber, 30% starch and 16.5% crude protein; forages were corn silage (67% of forage dry matter) and alfalfa silage (33% of forage dry matter). Concentrations of B vitamins were analyzed in feed and duodenal digesta. Apparent ruminal synthesis of each B vitamin was calculated as the duodenal flow minus the intake. Under the present experimental conditions, a dietary supplement of Saccharomyces cerevisiae fermentation product had no effect apparent synthesis of B vitamins in the rumen or on the amounts of these vitamins reaching the duodenum and available for absorption by the dairy cow., (Crown Copyright © 2017 Published by FASS and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

This study was undertaken to evaluate the effect of supplementation of folic acid and vitamin B 12 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 12 , or (4) 320 mg of folic acid and 10 mg of vitamin B 12 . 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- 2 H 2 ]-glucose (16.5 mmol/h; jugular vein) and [1- 13 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 12 respectively increased folate and vitamin B 12 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 12 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 12 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., (Crown Copyright © 2017 Published by FASS and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

The purpose of this experiment was to gain understanding on changes in energy partitioning when folic acid and vitamin B 12 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 12 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 12 supplement increased plasma concentration of vitamin B 12 . 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 12 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., (Crown Copyright © 2017 Published by FASS and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

Effects of plant maturity on apparent ruminal synthesis and post-ruminal supply of B vitamins were evaluated in two feeding trials. Diets containing alfalfa (Trial 1) or orchardgrass (Trial 2) silages harvested either (1) early cut, less mature (EC) or (2) late cut, more mature (LC) as the sole forage were offered to ruminally and duodenally cannulated lactating Holstein cows in crossover design experiments. In Trial 1, conducted with 16 cows (569±43 kg of empty BW (ruminal content removed) and 43.7±8.6 kg/day of 3.5% fat-corrected milk yield; mean±SD) in two 17-day treatment periods, both diets provided ~22% forage NDF and 27% total NDF, and the forage-to-concentrate ratios were 53 : 47 and 42 : 58 for EC and LC, respectively. In Trial 2, conducted with 13 cows (588±55 kg of empty BW and 43.7±7.7 kg/day of 3.5% fat-corrected milk yield; mean±SD) in two 18-day treatment periods, both diets provided ~25% forage NDF and 31% total NDF; the forage-to-concentrate ratios were 58 : 42 and 46 : 54 for EC and LC, respectively. Thiamin, riboflavin, niacin, vitamin B6, folates and vitamin B12 were measured in feed and duodenal content. Apparent ruminal synthesis was calculated as the duodenal flow minus the intake. Diets based on EC alfalfa decreased the amounts of thiamin, niacin and folates reaching the duodenum, whereas diets based on EC orchardgrass increased riboflavin duodenal flow. Daily apparent ruminal synthesis of thiamin, riboflavin, niacin and vitamin B6 were correlated negatively with their intake, suggesting a microbial regulation of their concentration in the rumen. Vitamin B12 apparent ruminal synthesis was correlated negatively with total volatile fatty acids concentration, but positively with ruminal pH and microbial N duodenal flow.

Effects of the forage-to-concentrate ratio on apparent ruminal synthesis of thiamine, riboflavin, niacin, vitamin B 6 , folates, and vitamin B 12 were evaluated in an experiment using 14 ruminally and duodenally cannulated Holstein cows. The experiment was a crossover design with two 15-d treatment periods and a 14-d preliminary period in which cows were fed a diet intermediate in composition between the treatment diets. Treatments were diets containing low-forage (44.8% forage, 32.8% starch, 24.4% neutral detergent fiber) or high-forage (61.4% forage, 22.5% starch, 30.7% neutral detergent fiber) concentrations. Both diets were formulated with different proportions of the same ingredients. Concentrations of B vitamins were analyzed in feed and duodenal digesta. Apparent ruminal synthesis of each B vitamin was calculated as the duodenal flow minus the intake. The high-forage diet had the highest concentrations of riboflavin, niacin, vitamin B 6 , and folates, whereas the low-forage diet had the highest thiamine concentration. Vitamin B 12 in the diets was under the level of detection. Consequently, despite a reduction in dry matter intake when the cows were fed the high-forage diet, increasing dietary forage concentration increased or tended to increase intakes of riboflavin, niacin, and vitamin B 6 but reduced thiamine and folate intakes. Increasing dietary forage concentration reduced apparent ruminal degradation of thiamine and apparent ruminal synthesis of riboflavin, niacin, and folates and increased ruminal degradation of vitamin B 6 , but had no effect on ruminal synthesis of vitamin B 12 . As a consequence, increasing the forage-to-concentrate ratio had no effect on the amounts of thiamine, riboflavin, and vitamin B 12 reaching the small intestine but decreased the amounts of niacin, vitamin B 6 , and folates available for absorption. Apparent ruminal syntheses of riboflavin, niacin, folates, and vitamin B 12 were correlated positively with the amount of starch digested in the rumen and duodenal flow of microbial N, whereas these correlations were negative for thiamine. Apparent ruminal syntheses of thiamine and vitamin B 6 were negatively correlated with their respective intakes, whereas folate intake was positively correlated with its synthesis in the rumen., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The metabolites of choline have a central role in many mammalian biological processes, and choline supplementation to the periparturient dairy cow improves hepatic lipid metabolism. However, variability in responses to choline supplementation has highlighted a lack of understanding of choline absorption in the lactating dairy cow. Our objective was to determine net choline absorption by measuring net portal fluxes of choline and choline metabolites in cows receiving either dietary supplements of rumen-protected choline (RPC) or abomasal delivery of choline (ADC). We also evaluated markers for choline bioavailability by examining relationships between net portal absorption of choline and choline metabolites in plasma and milk. Five late-lactation Holstein cows were used in a 5×5 Latin square design, with 5-d treatment periods and a 2-d interval between periods. Treatments were (1) control (0g/d of choline), (2) 12.5g/d of choline fed as RPC, (3) 25g/d of choline fed as RPC, (4) 12.5g/d of choline provided as ADC, and (5) 25g/d of choline provided as ADC. At the end of each 5-d period, milk was sampled and 9 blood samples were collected simultaneously from an artery and portal vein at 30-min intervals. Plasma, milk, and feed ingredient concentrations of acetylcholine, betaine, free choline, glycerophosphocholine, lysophosphatidylcholine, phosphatidylcholine, phosphocholine, and sphingomyelin were quantified by hydrophilic interaction liquid chromatography-tandem mass spectrometry. With an increasing dose of ADC, the net portal flux of free choline increased and regression analysis indicated 61% net absorption of the infused dose. Among the choline metabolites, only concentrations of betaine, free choline, and phosphocholine increased in both arterial plasma (3.9, 1.9, and 0.4 times, respectively) and milk (2.5, 1.4, and 1.0 times, respectively) with 25g/d of ADC relative to the control. For RPC, the net portal flux of free choline was low relative to ADC (13%), which was similar to the relative difference observed in the concentrations and yields of milk free choline and betaine (averaged 21%). When evaluating markers for choline bioavailability, betaine was the leading candidate. Betaine in plasma and milk (alone or in combination with phosphocholine) was strongly associated with net free choline portal flux (coefficient of determination ranging from 0.64 to 0.79). In summary, free choline supply to the lactating dairy cow increases only specific choline metabolites in plasma and milk, which can be potential markers for choline bioavailability., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: A combined supplement of vitamins B9 and B12 was reported to increase milk and milk component yields of dairy cows without effect on feed intake. The present study was undertaken to verify whether this supplementation positively modifies the pathways involved in milk and milk component synthesis. Thus, by studying the transcriptome activity in these tissues, the effect of supplements of both vitamins on the metabolism of both liver and mammary gland, was investigated. For this study, 24 multiparous Holstein dairy cows were assigned to 6 blocks of 4 animals each according to previous 305-day milk production. Within each block, cows were randomly assigned to weekly intramuscular injections of 5 mL of either saline 0.9 % NaCl, 320 mg of vitamin B9, 10 mg of vitamin B12 or a combination of both vitamins (B9 + B12). The experimental period began 3 weeks before the expected calving date and lasted 9 weeks of lactation. Liver and mammary biopsies were performed on lactating dairy cows 64 ± 3 days after calving. Samples from both tissues were analyzed by microarray and qPCR to identify genes differentially expressed in hepatic and mammary tissues., Results: Microarray analysis identified 47 genes in hepatic tissue and 16 genes in the mammary gland whose expression was modified by the vitamin supplements. Gene ontology (GO) categorizes genes in non-overlapping domains of molecular biology. Panther is one of the online GO resources used for gene function classification. It classifies the 63 genes according to Molecular Function, Biological Process and Protein Class. Most of the biological processes modulated by the vitamin supplements were associated to developmental process, protein metabolic process, transport and response to inflammation. In the liver, most of the genes modulated by the vitamin treatments involved protein metabolic process while developmental process appeared to be more affected by the treatments in mammary gland. Out of 25 genes analysed by qPCR, 7 were validated., Conclusion: The results indicate that several metabolic processes were modulated by the supplementation of vitamins in early-lactating dairy cows. In addition, the results suggest that the vitamin supplements promoted liver regeneration and reduced catabolism of lipids in early lactation.

Effects of particle length of silages on apparent ruminal synthesis (ARS) and postruminal supply of B vitamins were evaluated in 2 feeding trials. Diets containing alfalfa (trial 1) or orchardgrass (trial 2) silages, chopped to either 19mm (long cut, LC) or 10mm (short cut, SC) theoretical particle length, as the sole forage were offered to ruminally and duodenally cannulated lactating Holstein cows in crossover design experiments. Forages chopped to a theoretical particle length of 19 and 10mm had mean particles sizes of 14.1 and 8.1mm, respectively, in trial 1, and 15.3 and 11.3mm, respectively, in trial 2. Trial 1 was conducted with 13 multiparous cows in two 19-d treatment periods; both diets contained approximately 20% forage neutral detergent fiber (NDF), 25% total NDF, and forage-to-concentrate ratios were approximately 47:53. Trial 2 was conducted with 15 cows in two 18-d treatment periods; both diets contained approximately 23% forage NDF, 28% total NDF, and had a forage-to-concentrate ratio of 50:50. Thiamine, riboflavin, niacin, vitamin B6, folates, and vitamin B12 were measured in feed and duodenal content. Daily ARS was calculated as the duodenal flow minus the intake. In trial 1, daily intake of individual B vitamins was increased with the LC diet, but ARS of thiamine, riboflavin, vitamin B6, and folates was reduced. In trial 2, except for folates, intakes of the other B vitamins were decreased with the LC diets, whereas ARS of riboflavin, niacin, and vitamin B6 was increased. Daily ARS of thiamine, riboflavin, niacin, and vitamin B6 were correlated negatively with their intake, suggesting that ruminal bacteria reduced their synthesis when dietary supply increased. Microbial activity could have also reduced degradation of thiamine, riboflavin, and niacin, which is supported by (1) the negative correlation between ARS of these vitamins and ruminal pH or microbial N duodenal flow; and (2) the positive correlation between ARS and ruminal concentrations of volatile fatty acids. Folate ARS followed the opposite correlation pattern. Nevertheless, in spite of differences in intake and ARS, with both forages, decreasing particle length of silages had limited effects on the amounts of B vitamins reaching the sites of absorption in the small intestine of dairy cows., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Effects of nitrogen level and carbohydrate source on apparent ruminal synthesis (ARS) of thiamin, riboflavin, niacin, vitamin B6, folates, and vitamin B12 were evaluated using 4 lactating Holstein cows distributed in a 4 × 4 Latin square design with treatments following a 2 × 2 factorial arrangement. Cows were fitted with cannulas in the rumen and proximal duodenum. The treatments were 2 N levels and 2 carbohydrate sources. The diet with the high N level provided 14% crude protein, calculated to meet 110% of the protein requirements and an adequate supply in rumen-degradable protein, whereas the diet with the low N level contained 11% crude protein, calculated to meet 80% of the protein requirements with a shortage in rumen-degradable protein. Carbohydrate source treatments differed by their nature (i.e., high in starch from barley, corn, and wheat, or high in fiber from soybean hulls and dehydrated beet pulp). All 4 diets were isoenergetic, based on corn silage, and had the same forage-to-concentrate ratio (60:40, dry matter basis). Duodenal flow was determined using YbCl3 as a marker. Each B-vitamin ARS was calculated as duodenal flow minus daily intake. The intake of several B vitamins varied among treatments, but because the animals consumed a similar amount of feed every day (average of 20 kg of dry matter/d) the difference was mostly due to vitamin content of each ingredient and their relative proportion in the diets. Decreasing N concentration in the diet reduced vitamin B6 duodenal flow and increased its apparent ruminal degradation. It also decreased duodenal flow and ARS of folates. The high-starch diets increased duodenal flow and ruminal balance of riboflavin, vitamin B6, and folates, whereas the high-fiber diets increased vitamin B12 ARS and duodenal flow. These effects on apparent synthesis are possibly due to changes in ruminal fermentation., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Effects of forage family (legume vs. grass) on apparent ruminal synthesis (ARS) and postruminal supply of B vitamins were evaluated in 2 experiments. Diets containing either alfalfa (AL) or orchardgrass (OG) silages as the sole forage were offered to ruminally and duodenally cannulated lactating Holstein cows in crossover design experiments. Experiment 1 compared diets containing AL and OG [~23% forage neutral detergent fiber (NDF) and ~27% total NDF] offered to 8 cows in two 15-d treatment periods. Experiment 2 compared diets containing AL and OG (~25% forage NDF and ~30% total NDF) offered to 13 cows in two 18-d treatment periods. Thiamin, riboflavin, niacin, vitamin B6, folates, and vitamin B12 were analyzed in feeds and duodenal digesta. Apparent ruminal synthesis was calculated as the duodenal flow of each vitamin minus its intake. Forage family affected B vitamin intakes, duodenal flow, and ARS. In both experiments, AL diets increased vitamin B6 and decreased folate intakes. In experiment 1, riboflavin and niacin intakes were greater with the OG diet, whereas in experiment 2 thiamin intake was greater but riboflavin intake was smaller with the OG diet. In spite of the low contribution of either silage to the dietary folate content, folate intake was greater with OG diets than AL due to the difference in soybean meal contribution between diets. Niacin and folate ARS were not affected by the forage family. Duodenal microbial nitrogen flow was positively correlated with ARS of riboflavin, niacin, vitamin B6, folates, and vitamin B12, but tended to be negatively correlated with thiamin ARS. Apparent ruminal synthesis of folates and vitamin B12 appear to be related to microbial biomass activity. Changes in nutrient composition of the diets likely affected the microbial population in the rumen and their B vitamin metabolism., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Bioavailability of vitamin B12 is low in humans and animals. Improving vitamin B12 absorption is important for optimal performance in dairy cows and for increasing vitamin B12 concentrations in milk for human consumption. However, when supplemented in the diet, 80% of synthetic vitamin B12, cyanocobalamin (CN-CBL), is degraded in the rumen of dairy cows and only 25% of the amount escaping destruction in the rumen disappears from the small intestine between the duodenal and ileal cannulas. In pigs, vitamin B12 from milk is more efficiently absorbed than synthetic CN-CBL. The objective of this study was to determine the efficacy of casein hydrolysate and whey proteins as excipients for CN-CBL to increase portal-drained viscera (PDV) flux of the vitamin in lactating dairy cows. Four multiparous lactating Holstein cows (237 ± 17 DIM) equipped with a rumen cannula and catheters in the portal vein and a mesenteric artery were used in a randomized Youden square design. They were fed every 2 h to maintain steady digesta flow. On experimental days, they received a postruminal bolus of (1) CN-CBL alone (0.1 g), (2) CN-CBL (0.1 g) + casein hydrolysate (10 g), or (3) CN-CBL (0.1 g) + whey proteins (10 g). Starting 30 min after the bolus, blood samples were taken simultaneously from the 2 catheters every 15 min during the first 2 h and then every 2 h until 24 h postbolus. Milk yield, DMI, and vitamin B12 portal-arterial difference and PDV flux were analyzed using the MIXED procedure of SAS. Milk yield and DMI were not affected by treatments. The portal-arterial difference of vitamin B12 during the 24-h period following the bolus of vitamin was greater when the vitamin was given in solution with casein hydrolysate (2.9 ± 4.6 pg/mL) than alone (-17.5 ± 5.2 pg/mL) or with whey protein (-13.4 ± 4.2 pg/mL). The treatment effects were similar for the PDV flux. The present results suggest that CN-CBL given with casein hydrolysate increases vitamin B12 absorption as compared with CN-CBL given alone., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The fertility of dairy cows is challenged during early lactation, and better nutritional strategies need to be developed to address this issue. Combined supplementation of folic acid and vitamin B12 improve energy metabolism in the dairy cow during early lactation. Therefore, the present study was undertaken to explore the effects of this supplement on gene expression in granulosa cells from the dominant follicle during the postpartum period. Multiparous Holstein cows received weekly intramuscular injection of 320 mg of folic acid and 10 mg of vitamin B12 (treated group) beginning 24 (standard deviation=4) d before calving until 56 d after calving, whereas the control group received saline. The urea plasma concentration was significantly decreased during the precalving period, and the concentration of both folate and vitamin B12 were increased in treated animals. Milk production and dry matter intake were not significantly different between the 2 groups. Plasma concentrations of folates and vitamin B12 were increased in treated animals. Daily dry matter intake was not significantly different between the 2 groups before [13.5 kg; standard error (SE)=0.5] and after (23.6 kg; SE=0.9) calving. Average energy-corrected milk tended to be greater in vitamin-treated cows, 39.7 (SE=1.4) and 38.1 (SE=1.3) kg/d for treated and control cows, respectively. After calving, average plasma concentration of β-hydroxybutyrate tended to be lower in cows injected with the vitamin supplement, 0.47 (SE=0.04) versus 0.55 (SE=0.03) for treated and control cows, respectively. The ovarian follicle ≥12 mm in diameter was collected by ovum pick-up after estrus synchronization. Recovered follicular fluid volumes were greater in the vitamin-treated group. A microarray platform was used to investigate the effect of treatment on gene expression of granulosa cells. Lower expression of genes involved in the cell cycle and higher expression of genes associated with granulosa cell differentiation before ovulation were observed. Selected candidate genes were analyzed by reverse transcription quantitative PCR. Although the effects of intramuscular injections of folic acid and vitamin B12 on lactational performance and metabolic status of animals were limited, ingenuity pathway analysis of gene expression in granulosa cells suggests a stimulation of cell differentiation in vitamin-treated cows, which may be the result of an increase in LH secretion., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Homocysteine (Hcy) is an intermediary sulphur amino acid recognised for pro-oxidative properties in several species which may weaken immune competence in piglets. In this species, there is an acute 10-fold increase of concentrations of plasma Hcy (pHcy) during the first 2 weeks of life. The present experiment aimed to determine if pHcy in piglets can be regulated by oral supplementations of betaine as a methyl group supplier, creatine for reducing the demand for methyl groups, choline with both previous functions and vitamin B6 as enzymic co-factor for Hcy catabolism. A total of seventeen sows (second parity) were fed gestation and lactation diets supplemented with folic acid (10 mg/kg) and vitamin B12 (150 µg/kg). Eight piglets in each litter received daily one of the eight following oral treatments (mg/kg body weight): (1) control (saline); (2) betaine (50); (3) choline (70); (4) creatine (300); (5) pyridoxine (0·2); (6) treatments 2 and 5; (7) treatments 3 and 4; and (8) treatments 2, 3, 4 and 5. According to age, pHcy increased sharply from 2·48 µm at birth to 17·96 µm at 21 d of age (P < 0·01). Concentrations of pHcy tended to be lower (P = 0·09) in treated than in control piglets but the highest and sole pairwise significant decrease (23 %) was observed between treatments 1 and 8 (P = 0·03). Growth from birth to 21 d of age was not influenced by treatments (P > 0·70). Therefore, it appears possible to reduce pHcy concentrations in suckling piglets but a combination of all chosen nutrients is required.

This study was undertaken to evaluate folate and vitamin B12 concentrations of colostrum and milk in early lactation of dairy cows fed different levels of energy during the dry period. A total of 84 multiparous Holstein cows were assigned to one of the following dietary treatments fed as a total mixed ration 57 d before the expected calving date: (1) high-energy one-group dry cow diet [1.35 Mcal of net energy for maintenance/kg of dry matter (DM); 56% corn silage, 12% wheat straw, and 32% concentrate mix on a daily DM basis]; (2) controlled-energy one-group dry cow diet (1.14 Mcal of net energy for maintenance/kg of DM; 29% corn silage, 36% wheat straw, and 35% concentrate mix on a daily DM basis); or (3) an intermediate step-up diet (controlled-energy diet from dry off until 29 d before the expected calving date and then switching to a diet representing a 50:50 blend of the controlled- and high-energy diets from 28 d before expected calving date until parturition; 1.24 Mcal of net energy for maintenance/kg of DM). After calving, all cows were fed the same diet served as a total mixed ration (44% corn silage, 14% grass silage, and 42% concentrate mix on a daily DM basis) until 42 d in milk (DIM). Colostrum samples were taken at the first milking after parturition and milk samples were taken during the morning milking at 11 and 39±2 DIM. Colostrum from the first milking and milk yields were weighed on the day of sampling. Colostrum yield from the first milking postpartum and milk yields at 11 and 39 DIM were unaffected by treatments. Colostrum yield averaged 6.8±0.7mg at the first milking postpartum, whereas milk yields at 11 and 39 DIM were, on average, 40.3±1.5 and 48.9±1.3mg/d, respectively. Folate concentrations in colostrum and milk were not different among treatments. Folate concentration of colostrum (440.3±18.8ng/mL) was higher than folate concentration in milk at 11 (93.7±3.0ng/mL) and at 39 DIM (78.4±2.6ng/mL). Vitamin B12 concentration in colostrum was higher for controlled-energy cows (31.7±1.4ng/mL) than intermediate cows (23.5±1.4ng/mL), whereas no treatment effect was noted for vitamin B12 concentration in milk. At 11 and 39 DIM, milk concentrations of vitamin B12 averaged 3.8 and 3.2±1.4ng/mL, respectively. In summary, results suggest that dietary change during the dry period could modify vitamin B12 concentration in colostrum, but had no effect on milk concentration of folates and vitamin B12 during early lactation., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Homocysteine (Hcy), an intermediary sulfur AA, is recognized as a powerful prooxidant with deleterious effects on physiological and immune functions. In piglets, there is an acute 10-fold increase of plasma concentrations of homocysteine (pHcy) during the first 2 wk of life. This project aimed to maximize pHcy variations within physiological ranges using typical supplies of folates and vitamin B12 (B12) to sows and piglets. Growth, immune response, and Hcy metabolism of piglets were studied until piglets reached 56 d of age. Third-parity sows were randomly assigned to a 2 × 2 split-plot design with 2 dietary treatments during gestation and lactation, S(-) (1 mg/kg folates and 20 µg/kg B12, n = 15) and S(+) (10-fold S(-) levels, n = 16), and 2 treatments to piglets within each half litter, intramuscular injections (150 µg) of B12 (P(+)) at d 1 and 21 (weaning) and saline (P(-)). Within each litter of 12 piglets, 3 P(+) and 3 P(-) piglets were studied for growth and Hcy metabolism, and the others were studied for immune responses. During lactation, plasma B12 decreased and was transiently greater in S(+) vs. S(-) piglets on d 1 and P(+) vs. P(-) piglets on d 7 (sow treatment × age and piglet treatment × age; P < 0.05). From 14 to 21 d of age, pHcy was 33% lower in S(+)P(+) vs. S(-)P(-) piglets (sow treatment × piglet treatment interaction; P < 0.05). At 56 d of age, hepatic B12 was greater and pHcy was lower for P(+) vs. P(-) piglets (P < 0.05). No treatment effect was observed on growth except for a lower postweaning G:F in S(+)P(-) piglets than in others (sow treatment × piglet treatment interaction; P < 0.05). Positive correlations were observed between pHcy and growth (r > 0.29, P < 0.05) before and after weaning. Antibody responses to ovalbumin and serum tumor necrosis factor-α were not affected by treatments, but postweaning serum IL-8 peaked earlier in S(-)P(-) vs. S(+)P(+) piglets (piglet treatment × age; sow treatment × piglet treatment interaction, P < 0.05). Proliferation of lymphocytes in response to the mitogen concanavalin A tended to be lower in culture media supplemented with sera from S(-) vs. S(+) piglets (P = 0.081) and P(-) vs. P(+) piglets (P = 0.098), and the reduction of response was more marked (P < 0.05) with high (>21 µM) compared to medium (17 to 21 µM) or low (<17 µM) pHcy. In conclusion, the present vitamin supplements to sows and/or piglets produced variations of pHcy that were not apparently harmful for growth performance of piglets. The greater pHcy, particularly prevalent in S(-) and/or P(-) piglets, had negative effects on some indicators of immune responses, suggesting that these young animals may be immunologically more fragile.