Vitamin B12 Concentration In Milk Research Articles

A comprehensive integration of factors affecting vitamin B12 concentration in milk of Holstein cows: genetic variability, milk productivity, animal characteristics, and feeding management

Daily vitamin B12 (VB12) requirements of humans can naturally be fulfilled by animal product consumption, especially products from ruminants because of bacteria dwelling in their rumen. Indeed, only bacteria can synthesize this vitamin. Milk is hence an excellent source of VB12. This cross-sectional study was undertaken to unravel factors, such as genetic variation, diet and cow characteristics, and milk production, explaining the large variation in milk B12 concentration among cows by using an integrative approach. Milk samples from 2 consecutive milkings were collected from 3,533 Canadian Holstein cows (1,239 first, 932 s, and 1,362 third and more lactations) located in 99 herds with various feeding management. For genetic variation analysis purpose, pedigrees were traced back for 3 complete generations for each sire and dam. A total of 10,021 identities were used in the subsequent genetic analyses. Milk VB12 averaged 4.2 ng/mL with a range between 0.7 and 9.0 ng/mL. Dietary fiber (NDF from forage, dietary NDF, ADF, and lignin) increased and dietary components related to energy (non-fiber carbohydrate, starch, net energy of lactation, and percentage of concentrate) decreased VB12 in milk. Milk VB12 varied with days in milk, with a similar pattern as milk fat and protein concentration lactation curves. Milk VB12 increased as age at calving increased. When disregarding the herd variance, heritability value was 0.37, meaning that milk VB12 can be modified by genetic selection. The final model including factors related to the diet, animal characteristics and milk productivity, and genetic variation explained 79% (pseudo-R2) of the milk VB12 variation. When excluding the random effect of the cow, i.e., excluding the animal and genetic relationships, the pseudo-R2 dropped to 43%, reinforcing the importance of genetic variation in explaining milk VB12 variation. To our knowledge, the present study is the most comprehensive evaluation of factors affecting milk VB12 variation including the greatest number of cows from various lactation stages.

Vitamin B12 is Low in Milk of Early Postpartum Women in Urban Tanzania, and was not Significantly Increased by High dose Supplementation.

The effect of maternal multivitamin supplementation on breast milk vitamin B12 concentrations has not been examined in Tanzania, where the prevalence of maternal plasma B12 insufficiency is 25.6%. Multivitamins (containing 50 µg vitamin B12) or placebo were provided during pregnancy and in the postpartum period. Breast milk samples were collected at or around six weeks postpartum from 491 participants in a trial of multivitamins (NCT00197548). Linear and logistic regression models were used to examine the effect of supplements on vitamin B12 concentration in milk and its associations with other variables including potential confounders. Median vitamin B12 concentration in breast milk was 206 pmol/L and 70% of women had levels indicating inadequacy (<310 pmol/L). Multivitamin supplements did not significantly reduce the odds of inadequate vitamin B12 in breast milk, suggesting suboptimal absorption. A single unit increase in maternal hemoglobin at six weeks was associated with 18% lower odds of inadequate vitamin B12 in breast milk. Participants with higher BMI at baseline had double the odds of having inadequate vitamin B12 than the reference group (<22 kg/m2). Trials to determine the optimal dose, route, and duration of supplementation to improve maternal B12 status in Sub-Saharan Africa are of utmost importance.

Vitamin B12 concentrations in milk from Norwegian women during the six first months of lactation

BackgroundHuman milk vitamin B12 (B12) concentrations depend on maternal status and intake; only few data are available in high-income countries.ObjectiveWe assessed human milk B12 concentrations during the first 6 months postpartum in Norwegian women and its association with maternal dietary B12 intake and maternal urinary methylmalonic acid (MMA) concentration.MethodsIn this cross-sectional study, 175 mothers, exclusively (80%) or partially (20%) breastfeeding, were included. Milk B12 was measured by IMMULITE®/IMMULITE® 1000 B12 competitive protein binding assay and urinary MMA relative to creatinine (MMA/Cr) by liquid chromatography–tandem-mass spectrometry. Maternal habitual B12 intake and supplement use were estimated using a food frequency questionnaire.ResultsMean human milk B12 concentration was 327 pmol/L (range 140–1089), with 402 pmol/L at 1 month (n = 21), 333 pmol/L at four months (n = 32), and 299 pmol/L at 6 months (n = 21). Maternal B12 intake was 5 µg/d, 89% met the Estimated Average Requirement, and supplement use did not affect milk B12 concentrations. MMA/Cr was low in all women compared with published data. In exclusively breastfeeding women, MMA/Cr (beta (95% CI) −42.5 (−82.5, −2.5) and time since birth (−4.9 (−9.6, −0.3)) were significant predictors of human milk B12 concentrations. There was no association between total B12 intake and milk B12 concentration or between total B12 intake and MMA/Cr.ConclusionsMaternal B12 status and human milk B12 concentrations are likely sufficient, based on adequate maternal B12 dietary intake combined with low urinary MMA concentrations. Nevertheless, milk B12 concentration fell during 6 months postpartum while maternal B12 status did not change.

Impact of diet management and composition on vitamin B12 concentration in milk of Holstein cows

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.

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

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

The Effect of Maternal Supplementation of Zinc, Selenium, and Cobalt as Slow-Release Ruminal Bolus in Late Pregnancy on Some Blood Metabolites and Performance of Ewes and Their Lambs.

The aim of this study was to determine the effect of the supplementation of a slow-release bolus of zinc (Zn), selenium (Se), and cobalt (Co) at late gestation (6week prepartum) on performance and some blood metabolites of Mehraban ewes and their lambs until weaning. Seventy pregnant ewes, 6weeks prior to expected lambing, were randomly divided into two groups (35 heads each) including (1) control group and (2) slow-release bolus group. Blood samples of ewes were obtained on day 10 prepartum and 45 and 90days postpartum, and milk samples were collected on day 45. Blood samples of lambs were collected on days 10, 45, and 90. Body weight at birth and weaning and average daily gain were higher and percentage of mortality and white muscle disease rate were lower in lambs whose mothers were given a bolus (P < 0.05). Slow-release bolus administration increased serum alkaline phosphatase and whole blood glutathione peroxidase activity, plasma concentrations of Zn, Se, and vitamin B12 in ewes and their lambs (P < 0.05). In addition, serum creatine phosphokinase activity of lambs whose mothers were given bolus was lower (P < 0.05). Serum concentration of T3 in bolus given ewes and their lambs was higher (P < 0.05) and serum T4 concentration was lower (P < 0.05). Zinc, Se, and vitamin B12 concentrations in milk were significantly higher in treated ewes (P < 0.05). Obtained results showed that maternal supplementation of zinc, selenium, and cobalt as slow-release ruminal bolus in late pregnancy improved some mineral status of ewes and their lambs until weaning and led to higher body weights of lambs at weaning.

Short communication: Factors affecting vitamin B12 concentration in milk of commercial dairy herds: An exploratory study

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.

Short communication: Casein hydrolysate and whey proteins as excipients for cyanocobalamin to increase intestinal absorption in the lactating dairy cow

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 2h to maintain steady digesta flow. On experimental days, they received a postruminal bolus of (1) CN-CBL alone (0.1g), (2) CN-CBL (0.1g) + casein hydrolysate (10g), or (3) CN-CBL (0.1g) + whey proteins (10g). Starting 30 min after the bolus, blood samples were taken simultaneously from the 2 catheters every 15 min during the first 2h and then every 2h until 24h 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.

Short communication: Folates and vitamin B12 in colostrum and milk from dairy cows fed different energy levels during the dry period

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.

Technical note: Development and validation of a method using ultra performance liquid chromatography coupled with tandem mass spectrometry for determination of vitamin B12 concentrations in milk and dairy products

A method using ultra performance liquid chromatography coupled with tandem mass spectrometry was developed to measure cobalamins in naturally enriched raw milk and to evaluate their fate during thermal treatments and along the process of cheese making. After addition of methotrexate as internal standard, samples were submitted to heat treatment in the presence of cyanide, which converts all the less-stable cobalamins into cyanocobalamin; then, purification was performed by a solid-phase extraction step. Reverse-phase ultra performance liquid chromatography separation coupled with tandem mass spectrometry provided a fast and reliable determination. Mass spectrometric analysis was carried out in multiple reaction monitoring mode. The monitored transitions were m/z 678.36 → 147.10 and 678.36 → 359.30 for vitamin B12 and m/z 455.22 → 175.13 and 455.22 → 308.22 for methotrexate (internal standard). The limit of quantification was 2ng/g. The method showed good linearity from 2 to 20ng/g (R2 ≥0.98) and intra- and interday precisions were always less than 19%.

Effects of cobalt supplementation and vitamin B12 injections on lactation performance and metabolism of Holstein dairy cows

The objective of this study was to determine lactation performance and metabolism of primiparous and multiparous dairy cows fed different levels and sources (inorganic and organic) of Co or given weekly vitamin B12 injections. Forty-five primi- and multiparous cows at 60d prepartum were blocked by expected calving date, and randomly assigned to 1 of 5 treatments in a randomized complete block design with treatments starting at 60d prepartum. The 5 treatments were (1) no supplemental dietary Co (control, CON), (2) 25mg/d of supplemental dietary Co from Co carbonate (CoCarb), (3) 25mg/d of supplemental dietary Co from Co glucoheptonate (LCoGH), (4) 75mg/d of supplemental dietary Co from Co glucoheptonate (HCoGH), and (5) CON diet plus weekly 10mg i.m. of vitamin B12 injections (IB12). Cows remained on their respective treatment until 150d after calving. Cobalt concentrations (mg/kg of dry matter) in the lactating diets were 1.0, 1.9, 2.3, and 5.1 for CON/IB12, CoCarb, LCoGH, and HCoGH, respectively. Dry matter intake, body weight, and body condition score were not affected by treatment. The LCoGH treatment tended to have greater milk yield than CoCarb, and CON had similar milk yields to the mean of LCoGH and HCoGH. Cobalt supplementation or the use of vitamin B12 injections did not influence plasma or liver measures of energy metabolism. Injections of vitamin B12 increased plasma, liver, and milk vitamin B12 contents. Dietary Co addition did not affect plasma vitamin B12 concentrations; however, it did increase milk vitamin B12 concentrations throughout lactation and liver vitamin B12 at calving with no effect of source or level of Co. Folate status of cows in the study was low and possibly limited the effect of improved vitamin B12 status on lactation performance. Overall, Co supplementation (inorganic and organic) or vitamin B12 injections improved measures of vitamin B12 status, but not lactation performance compared with CON possibly due to Co being above requirements in the CON diet.

Breast milk vitamin B12 concentrations in Guatemala: relationship to maternal and infant intake and status

There are no studies of breast milk vitamin B12 in groups with a range of B12 status, and previous methods are invalid due to failure to remove apohaptocorrin (apoHc) prior to analysis. B12 was measured in breast milk collected at 12 ± 0.5 mo postpartum from 129 women in a low income Guatemalan community. Complete data on maternal and infant plasma B12 and intakes were available for 83 mother‐infant pairs. Plasma B12 was measured (competitive protein‐binding radioassay) and cut‐offs designated Deficient (&lt;148 pmol/L), Marginal (148‐220 pmol/L) and Adequate (&gt;220 pmol/L) maternal and infant status. Milk B12 was analyzed by chemiluminescence after removing apoHc by cobinamide‐coated EAH sepharose gel. One third of women were B12 Deficient, one third had Marginal status, and 88% of infants were Deficient or Marginal. Both infant plasma and breast milk B12 were sig. higher in Adequate vs Deficient or Marginal status women. Breast milk B12 was below the detectable limit of 50 pmol/L in 47% of samples, none from Adequate mothers. In the Adequate group milk B12 averaged 128 pmol/L, vs. 870 pmol/L in 21 healthy Californian women, providing only ≈0.1 ug/d for exclusively breastfed infants. Maternal plasma B12 correlated with milk and infant plasma B12 (both r=0.36, P&lt;0.001), and milk correlated with maternal intake. Since high B12 doses to mothers did not improve infant status, interventions are needed before/during pregnancy.

The effect of increasing the Vitamin B12 status of Romney ewes on foetal liver Vitamin B12, milk Vitamin B12 and liver Vitamin B12 concentrations in suckling lambs

Aim. To determine the effect of increasing the Vitamin B12 status of the ewe on the Vitamin B12 supply to the suckling lamb. Methods. The Vitamin B12 status of the ewe was increased during gestation and lactation by three injections of a long- acting preparation of Vitamin B12 microencapsulated in an organic acid polymer. The Vitamin B12 status of the ewes and suckling lambs was assessed from changes in serum and liver Vitamin B12 concentrations. Results. Compared to untreated animals, serum and liver Vitamin B12 concentrations of the treated ewes were increased at least 70% during gestation. Foetal liver Vitamin B12 concentrations were increased 270%. Over the lactation, ewe serum and milk Vitamin B12 concentrations were increased at least 200% and 44%, respectively. The liver Vitamin B12 stores of the new born lambs from Vitamin B,,-treated ewes were depleted within 58 days. There were no significant differences in the serum vitamin B12 concentrations of suckling lambs from vitamin B12-treated and untreated ewes. Conclusion. Ewes with a high vitamin B12 status will ensure an adequate supply of vitamin B12 to their lambs for at least the first 30 days of life. Clinical significance. In flocks grazing Co-deficient pastures, treating ewes with a long-acting vitamin B12 supplement at mating will prevent Vitamin B12 (Co) deficiency in ewes, as well as their lambs, until they can be treated at tailing at 4-6 weeks of age.

Vitamin B12 responses to cobalt pellets in beef cows.

To assess the effectiveness of cobalt pellets in maintaining adequate vitamin B12 in beef cows on pasture of low cobalt content. A field experiment in a herd grazing cobalt deficient pasture. Mature Murray Grey cows. Cows were given a single oral dose of 0, 1, 2 or 4 cobalt pellets (30 g pellets containing 30% by weight cobaltic oxide) with a selenium pellet and a grub screw. Samples of blood, liver, faeces and milk for chemical analysis were collected at intervals over a period of 2 years after treatment. A single cobalt pellet raised liver vitamin B12 concentration of cows above that of untreated cows for at least 28 weeks, and 2 or 4 pellets for 57 weeks. Plasma vitamin B12 concentration was an unreliable indicator of the effectiveness of cobalt pellet therapy. Milk vitamin B12 and faecal cobalt concentrations increased in response to cobalt pellet therapy. These studies show that one cobalt pellet will prevent vitamin B12 inadequacy in beef cows for between 28 and 57 weeks; two or four pellets will prevent inadequacy for 57 to 75 weeks. Milk vitamin B12 concentration may be a useful indicator of the effectiveness of cobalt pellets in increasing the vitamin B12 supply in lactating cows.

The effects of vitamin C, vitamin B6, and vitamin B12 supplementation on the breast milk and maternal status of well-nourished women1–3

The effects of vitamin supplements and/or diet on the levels of vitamin C, vitamin B6, and vitamin B12 in milk and blood of lactating women were determined. At the end of gestation, subjects were divided into two lactation groups: supplemented (10 subjects) and nonsup-plemented (seven subjects). Milk samples were collected from 5 to 7 days and 43 to 45 days postpartum. Fasting blood samples were drawn at 8 and 46 days postpartum for vitamin C, B6, and B12 status measurements. Dietary records of all foods consumed by the subject were kept for 4 days at 1 and 6 weeks postpartum. The vitamin B6 level in breast milk of the unsupplemented group of mothers was significantly lower (P < 0.05) than the supplemented group of women at 5 to 7 days postpartum. Vitamin B12 concentration in milk of nonsupplemented mothers at 43 to 45 days postpartum was significantly lower (P < 0.05) than the supplemented group of women at 43 to 45 days postpartum. None of the milk values or the maternal blood levels measured in the women was less than published norms for vitamin C, vitamin B6, and vitamin B12.