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Effect of dietary supplements of biotin, intramuscular injections of vitamin B12, or both on postpartum lactation performance in multiparous dairy cows

  • D.M. Wang
    Affiliations
    Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R. China
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  • B.X. Zhang
    Affiliations
    Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R. China
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  • J.K. Wang
    Affiliations
    Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R. China
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  • H.Y. Liu
    Affiliations
    Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R. China
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  • J.X. Liu
    Correspondence
    Corresponding author
    Affiliations
    Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R. China
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Open ArchivePublished:July 11, 2018DOI:https://doi.org/10.3168/jds.2018-14524

      ABSTRACT

      The current study was conducted to investigate the effects of dietary supplementation of biotin, intramuscular injections of vitamin B12 (VB12), or both beginning at the prepartum period on feed intake and lactation performance in postpartum dairy cows. Forty-eight dairy cows were allocated into 12 blocks, based on parity and milk yield of the previous lactation cycle, and randomly assigned to 1 of 4 treatments. Supplementation of VB12 (weekly intramuscular injections of 0 or 10 mg) and biotin (dietary supplements of 0 or 30 mg/d) were used in a 2 × 2 factorial arrangement in a randomized complete block design of 12 blocks with repeated measures. The study started at 3 wk before the expected calving date and ended at 8 wk after calving. Feed intake and lactation performance (milk yield and composition) were recorded weekly after calving. Blood variables were measured on d −10, 0, 8, 15, 29, 43, and 57 relative to calving. When VB12 was given, the cows had greater feed intake, better lactation performance and lower body weight loss in the postpartum period compared with animals without injection of VB12. The VB12-injected cows had lower plasma nonesterified fatty acids and β-hydroxybutyrate concentrations but higher plasma superoxide dismutase activity compared with cows without VB12. Cows fed a biotin supplement had higher milk protein yield (6 and 8 wk) and lactose yield (6–8 wk), compared with animals without biotin. However, under the present experimental conditions, we found no additive effect of a combined supplement of biotin and vitamin B12 on lactation performance of dairy cows.

      Key words

      INTRODUCTION

      Hepatic gluconeogenesis plays an important role in net hepatic glucose generation (
      • Reynolds C.K.
      Production and metabolic effects of site of starch digestion in dairy cattle.
      ), and increasing hepatic gluconeogenesis is an efficient way to increase the milk yield in lactating dairy cows (
      • Rukkwamsuk T.
      • Wensing T.
      • Geelen M.J.
      Effect of fatty liver on hepatic gluconeogenesis in periparturient dairy cows.
      ;
      • Karcher E.L.
      • Pickett M.M.
      • Varga G.A.
      • Donkin S.S.
      Effect of dietary carbohydrate and monensin on expression of gluconeogenic enzymes in liver of transition dairy cows.
      ). In gluconeogenesis, the propionate absorbed by hepatic cells is transformed to methylmalonyl-CoA by propionyl-CoA carboxylase, a biotin-dependent enzyme (
      • Scott J.M.
      Folate and vitamin B12.
      ;
      • Hügler M.
      • Krieger R.S.
      • Jahn M.
      • Fuchs G.
      Characterization of acetyl-CoA/propionyl-CoA carboxylase in Metallosphaera sedula. Carboxylating enzyme in the 3-hydroxypropionate cycle for autotrophic carbon fixation.
      ), and further isomerized to succinyl-CoA by methylmalonyl-CoA mutase, a vitamin B12 (VB12)-dependent enzyme (
      • Padmakumar R.
      • Padmakumar R.
      • Banerjee R.
      Evidence that cobalt-carbon bond homolysis is coupled to hydrogen atom abstraction from substrate in methylmalonyl-CoA mutase.
      ). Thus, a beneficial effect would be expected when additional VB12 or biotin is provided to the dairy cows with an insufficient supply in VB12 or biotin during early lactation.
      The effect of biotin or vitamin B12 on milk production in lactating cows has been addressed in some studies. In early-lactation dairy cows, supplementary biotin could improve DMI and milk yield (
      • Zimmerly C.A.
      • Weiss W.P.
      Effects of supplemental dietary biotin on performance of Holstein cows during early lactation.
      ); however, this was not found in studies conducted by
      • Fitzgerald T.
      • Norton B.W.
      • Elliott R.
      • Podlich H.
      • Svendsen O.L.
      The influence of long-term supplementation with biotin on the prevention of lameness in pasture fed dairy cows.
      and
      • Ganjkhanlou M.
      • Salimi M.
      • Nikkhah A.
      • Zali A.
      Effects of supplemental dietary biotin on performance of Holstein dairy cows.
      . In terms of VB12,
      • Girard C.L.
      • Matte J.J.
      Effects of intramuscular injections of vitamin B12 on lactation performance of dairy cows fed dietary supplements of folic acid and rumen-protected methionine.
      found that intramuscular injection of VB12 alone tended to increase milk total solid contents in cows fed dietary supplements of folic acid, but in the report of
      • Akins M.S.
      • Bertics S.J.
      • Socha M.T.
      • Shaver R.D.
      Effects of cobalt supplementation and vitamin B12 injections on lactation performance and metabolism of Holstein dairy cows.
      intramuscular injection of VB12 did not improve lactation performance of dairy cows.
      It was noted that plasma biotin concentration of postpartum dairy cows was 50% of that in prepartum dairy cows (
      • Rosendo O.
      • Staples C.R.
      • Mcdowell L.R.
      • Mcmahon R.
      • Badinga L.
      • Martin F.G.
      • Shearer J.F.
      • Seymour W.M.
      • Wilkinson N.S.
      Effects of biotin supplementation on peripartum performance and metabolites of Holstein cows.
      ). Similarly, plasma VB12 concentration was at the lowest level during 0 to 60 DIM (
      • Girard C.L.
      • Matte J.J.
      Changes in serum concentrations of folates, pyridoxal, pyridoxal-5-phosphate and vitamin B12 during lactation of dairy cows fed dietary supplements of folic acid.
      ); these data suggest a shortage of VB12 and biotin in transition dairy cows. Although the independent addition of VB12 or biotin has been proven to be beneficial to early-lactating dairy cows (
      • Zimmerly C.A.
      • Weiss W.P.
      Effects of supplemental dietary biotin on performance of Holstein cows during early lactation.
      ;
      • Girard C.L.
      • Matte J.J.
      Effects of intramuscular injections of vitamin B12 on lactation performance of dairy cows fed dietary supplements of folic acid and rumen-protected methionine.
      ), it is not clear whether a combined (VB12 plus biotin) treatment beginning at the precalving stage would have an interactive effect on the lactation performance of transition cows. Thus, the current study was conducted to investigate the effects of dietary biotin/intramuscular VB12 injection, starting at the prepartum stage, on feed intake, milk performance, BW change, and plasma variables in the postpartum dairy cows.

      MATERIALS AND METHODS

       Animals, Diets, and Experimental Design

      This experiment was carried out at the Hangjiang Dairy Farm (Hangzhou, China). Animal use was approved by the Animal Care Committee of Zhejiang University. Forty-eight multiparous Holstein dairy cows (BW = 691 kg, SD = 9.9; parity = 2.75, SD = 0.10; milk yield of last lactation period = 31.2 kg/d, SD = 0.55) were allocated into 12 blocks based on the parity and milk yield of previous lactation period. The cows within each block were randomly assigned to 1 of 4 treatments—control, biotin, VB12, and biotin plus VB12—in addition to the basal TMR. Biotin (30 mg/d) was added to the cow ration 3 times daily during the feeding period. The VB12 (10 mg/wk) was administered by intramuscular injections (
      • Duplessis M.
      • Lapierre H.
      • Pellerin D.
      • Laforest J.P.
      • Girard C.L.
      Effects of intramuscular injections of folic acid, vitamin B12, or both, on lactational performance and energy status of multiparous dairy cows.
      ). The ingredients and chemical composition of pre- and postpartum TMR are presented in Table 1. Cows were housed in tiestall barns and had free access to fresh water during the experimental period. The TMR was provided to the cows daily at 0630, 1400, and 1900. The cows were milked 3 times daily during the feeding period. The experiment was conducted from 3 wk before expected calving to 8 wk after calving.
      Table 1Dietary ingredients and chemical composition of pre- and postpartum dairy cows
      ItemPrepartumPostpartum
      Ingredient, % of DM
       Corn grain, ground12.224.2
       Steam-flaked corn7.33
       Soybean meal10.813.4
       Barley3.913.19
       Cottonseed meal3.20
       Whole cottonseeds5.07
       Sugar beet pulp4.175.35
       Wheat bran2.44
       Corn silage17.49.94
       Alfalfa hay7.9522.9
       Chinese wild ryegrass24.1
       Oat grass11.54.91
       Premix
      Contained (per kilogram of premix): 250 KIU of vitamin A, 50 KIU of vitamin D, 2,300 IU of vitamin E, 600 mg of Fe, 650 mg of Cu, 3,000 mg of Zn, 630 mg of Mn, 17 mg of Se, 36 mg of I, 15–18% of NaCl, water ≤10%.
      2.143.65
      Composition, % of DM
       CP11.816.3
       NDF42.332.3
       ADF23.318.5
       Ca0.830.88
       P0.380.42
       Co,
      Co content in the premix.
      mg/kg
      1.702.90
       Ash7.527.94
       NEL, Mcal/kg of DM1.62
      1 Contained (per kilogram of premix): 250 KIU of vitamin A, 50 KIU of vitamin D, 2,300 IU of vitamin E, 600 mg of Fe, 650 mg of Cu, 3,000 mg of Zn, 630 mg of Mn, 17 mg of Se, 36 mg of I, 15–18% of NaCl, water ≤10%.
      2 Co content in the premix.
      The experiment was carried out along with another study (
      • Wang D.M.
      • Zhang B.X.
      • Wang J.K.
      • Liu H.Y.
      • Liu J.X.
      Short communication: Effects of dietary 5,6-dimethylbenzimidazole supplementation on vitamin B12 supply, lactation performance, and energy balance in dairy cows during the transition period and early lactation.
      ), and both experiments shared control animals. Therefore, the data for the control group were similar to those of
      • Wang D.M.
      • Zhang B.X.
      • Wang J.K.
      • Liu H.Y.
      • Liu J.X.
      Short communication: Effects of dietary 5,6-dimethylbenzimidazole supplementation on vitamin B12 supply, lactation performance, and energy balance in dairy cows during the transition period and early lactation.
      .

       DMI, Milk Production, and Milk Composition

      To calculate DMI, the feed offered and refused were weighed weekly for 3 consecutive days during the postpartum period. The representative samples of the TMR and orts were collected weekly for analysis of chemical composition. All samples were dried at 65°C for 48 h, ground through a 1-mm screen mesh using a high-speed grinder (Tecator 1093, Hoganas, Sweden), and stored in plastic bottles at 4°C to analyze the contents of DM (method no. 934.01), CP (method no. 955.04), ether extract (method no. 920.39), ash (method no. 942.05), and ADF (method no. 973.18) according to the
      • AOAC
      Official Methods of Analysis.
      . The NDF was analyzed by the method of
      • Van Soest P.J.
      • Robertson J.B.
      • Lewis B.A.
      Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition.
      .
      The individual milk yield was recorded on the same days as DMI determination using a milk-sampling device (Waikato Milking Systems NZ Ltd., Waikato, Hamilton, New Zealand). Milk samples collected from each cow on d 4 of each week were used to determine milk composition by using a Foss FT+ instrument (Foss Electric, Hillerød, Denmark). The milk samples collected at wk 8 were used to determine VB12 and biotin concentration. The collected milk samples were prepared according to the method described by
      • Indyk H.E.
      • Gill B.D.
      • Woollard D.C.
      Biotin content of paediatric formulae, early lactation milk and seasonal bovine milk powders by biosensor immunoassay.
      . In brief, 10.0 mL of milk was diluted with 40 mL of water and ultrasonicated. A 5-mL aliquot was placed in a 100°C water bath for 15 min and then centrifuged (4,500 × g, 15 min, 4°C;
      • Indyk H.E.
      • Gill B.D.
      • Woollard D.C.
      Biotin content of paediatric formulae, early lactation milk and seasonal bovine milk powders by biosensor immunoassay.
      ). Afterward, the supernatant was filtered through combined 0.45- and 0.22-µm filter paper for determination of VB12 by radioassay using a commercial kit (SimulTRAC B12/FOLATE-S, MP Biomedicals, Santa Ana, CA;
      • Duplessis M.
      • Lapierre H.
      • Pellerin D.
      • Laforest J.P.
      • Girard C.L.
      Effects of intramuscular injections of folic acid, vitamin B12, or both, on lactational performance and energy status of multiparous dairy cows.
      ); and biotin concentration by using another commercial kit (art. no. H1601, R-Biopharm GmbH, Darmstadt, Germany;
      • Chen B.
      • Wang C.
      • Liu J.X.
      Effects of dietary biotin supplementation on performance and hoof quality of Chinese Holstein dairy cows.
      ).

       Blood Sampling

      Blood samples (5 mL) were obtained from the coccygeal vein into evacuated tubes containing sodium heparin 4 h after the morning feeding on d −10, 0, 8, 15, 29, 43, and 57 relative to calving. Plasma was then harvested by centrifugation at 3,000 × g for 10 min at 4°C and stored at −20°C until analysis. Plasma samples were analyzed using an auto-analyzer 7020 (Hitachi High-Technologies Corp., Tokyo, Japan) for glucose, nonesterified fatty acids, BHB, BUN, and cholesterol, according to the method described in the previous study (
      • Wang D.M.
      • Wang C.
      • Liu H.Y.
      • Liu J.X.
      • Ferguson J.D.
      Effects of rumen-protected γ-aminobutyric acid on feed intake, lactation performance, and antioxidative status in early lactating dairy cows.
      ). The assay for plasma superoxide dismutase (SOD) activity was conducted using commercial kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China; A001–1,
      • Xiong J.L.
      • Wang Y.M.
      • Nennich T.D.
      • Li Y.
      • Liu J.X.
      Transfer of dietary aflatoxin B1 to milk aflatoxin M1 and effect of inclusion of adsorbent in the diet of dairy cows.
      ). Plasma sampled at d 57 were used to determine VB12 and biotin, with the same methods used for milk samples.

       BW and Energy Balance Status

      Animal BW was measured on d 0 and 57, relative to calving day, based on the measurement of body length and heart girth using the equation BW (kg) = heart girth2 (m) × body length (m) × 90 (
      • Yan T.
      • Mayne C.S.
      • Patterson D.C.
      • Agnew R.E.
      Prediction of body weight and empty body composition using body size measurements in lactating dairy cows.
      ). The energy balance (EB) status on wk 1 and 8 postpartum was determined as EB (Mcal/d) = (DMI × diet NEL) – [(0.08 × BW0.75) + (milk NEL × milk yield)] (
      • Spicer L.J.
      • Tucker W.B.
      • Adams G.D.
      Insulin-like growth factor-I in dairy cows: relationships among energy balance, body condition, ovarian activity, and estrous behavior.
      ).

       Statistical Analysis

      Data were analyzed using the MIXED procedure (
      • SAS Institute
      SAS User's Guide: Statistics. Version 8.01.
      ). Supplementation of VB12 (weekly intramuscular injections of 0 or 10 mg) and biotin (dietary supplements of 0 or 30 mg/d) were used in a 2 × 2 factorial arrangement in a 12 randomized complete block design with repeated measures. For milk production variables (DMI, yield and composition), covariance structure type AR(1) was used to fit equally spaced repeated measures. For plasma variables, covariance structure spatial power was applied to fit unequally spaced repeated measures (
      • Littell R.C.
      • Milliken G.A.
      • Stroup W.W.
      • Wolfinger R.D.
      SAS system for mixed models.
      ;
      • Chung Y.H.
      • Girard I.D.
      • Varga G.A.
      Effects of feeding dry propylene glycol to early postpartum Holstein dairy cows on production and blood parameters.
      ). The model included block, VB12, biotin, week, and the interactions of VB12 × biotin, VB12 × week, biotin × week, and VB12 × biotin × week as fixed effects. Cows within treatments were subjected as random variables. For all analyses, least squares means were calculated and differences between treatments were separated with a PDIFF option. Significant differences were established at P < 0.05 and trends at P ≤ 0.10 and P > 0.05.

      RESULTS

       Feed Intake, Lactation Performance, and BW Change

      Postpartum DMI was higher in cows with VB12 injection, compared with those not injected with VB12 (Table 2; P < 0.01). Compared with cows that were not treated with VB12, raw (P < 0.01) and energy-corrected (P = 0.05) milk yields were greater in cows with VB12 injection (Table 2). Overall, yields of protein and lactose were higher in VB12-injected cows compared with cows without injection of VB12 (P < 0.01; Table 2). Lactose content was higher in VB12 supplemented cows compared with cows that did not treated with VB12 (P < 0.01; Table 2). The VB12 content in the milk was greater in VB12-supplemented cows than in milk of animals without VB12-injection (P < 0.01; Table 2). No VB12 × week effect was observed on lactation performance of transition cows (P > 0.05). No effect of biotin on overall lactation performance was observed in postpartum dairy cows (P > 0.05). However, milk protein yield was higher in cows supplemented with biotin, compared with that of animals without biotin in wk 6 and 8 postpartum (biotin × week effect, P = 0.02; Figure 1A). Higher milk lactose yield was observed in cows fed biotin during wk 6 to 8 postpartum compared with the animals without no biotin supplemented (biotin × week, P = 0.05; Figure 1B). The milk biotin content was greater in biotin-supplemented cows than that of animals without biotin supplementation (P < 0.01, Table 2). The N conversion in the biotin-supplemented cows tended to be higher than the animals without biotin supplementation (P = 0.08). Supplementation of both VB12 and biotin had an interactive effect on improving milk protein content (P = 0.09).
      Table 2Effect of dietary biotin (Bio) and intramuscular injection of vitamin B12 (VB12) on DMI, lactation performance, and BW change during first 8 wk of lactation in dairy cows
      Sampling times: DMI and milk yield were recorded on d 3, 4, and 5 in each postpartum week, and milk components were determined on d 4 for each postpartum week. Biotin and VB12 concentrations were measured in the 8th week postpartum.
      ItemTreatmentSEMP-value
      ControlBioVB12Bio + VB12BioVB12Biotin + VB12
      DMI, kg/d21.421.123.122.80.580.29<0.010.75
      Milk yield, kg/d
       Raw36.437.239.539.70.990.66<0.010.82
       ECM
      ECM = 12.55 × fat yield (kg/d) + 7.39 × protein yield (kg/d) + 5.34 × lactose yield (kg/d), from Orth (1992).
      36.538.637.339.30.7880.500.050.95
       Fat1.451.511.481.490.0410.830.610.61
       Protein
      Biotin × week interaction, P ≤ 0.05.
      1.131.171.261.300.0250.88<0.010.47
       Lactose
      Biotin × week interaction, P ≤ 0.05.
      1.811.831.992.030.0650.30<0.010.74
      Content, g/100 g
       Fat4.084.183.843.970.1410.670.130.83
       Protein3.253.193.183.310.0380.560.610.09
       Lactose4.944.995.025.050.0250.300.040.15
       Biotin, ng/mL0.0940.280.1130.3190.0372<0.010.440.79
       VB12, ng/mL0.3300.4051.5432.4380.3900.23<0.010.31
      BW change, kg−42.9−37.5−32.0−29.64.240.370.030.73
      Energy balance, Mcal/d−4.07−3.75−2.28−2.011.160.290.840.99
      Feed efficiency
      Feed efficiency = milk yield/DMI.
      1.671.681.661.670.0430.970.990.86
      Nitrogen conversion
      Nitrogen conversion = milk protein yield/CP intake.
      0.310.310.300.320.090.080.510.40
      1 Sampling times: DMI and milk yield were recorded on d 3, 4, and 5 in each postpartum week, and milk components were determined on d 4 for each postpartum week. Biotin and VB12 concentrations were measured in the 8th week postpartum.
      2 ECM = 12.55 × fat yield (kg/d) + 7.39 × protein yield (kg/d) + 5.34 × lactose yield (kg/d), from
      • Orth R.
      Sample day and lactation report, DHIA 200. Fact Sheet A-2.
      .
      3 Biotin × week interaction, P ≤ 0.05.
      4 Feed efficiency = milk yield/DMI.
      5 Nitrogen conversion = milk protein yield/CP intake.
      Figure thumbnail gr1
      Figure 1Effects of dietary biotin [0 (dashed line) or 30 mg/d (solid line)] given from 3 wk before the expected calving date to 8 wk postpartum on milk protein yield (A, P = 0.02) and milk lactose yield (B, P = 0.05). Bars indicate the SEM; an asterisk (*) indicates a significant difference (P < 0.05) between treatments.
      The BW loss was lower in cows with VB12 injection (30.8 ± 2.92 kg), compared with animals without VB12 injection (40.2 ± 3.13 kg, P = 0.03; Table 2), with no interaction of VB12 and biotin on BW change (P > 0.05). We found no effect of VB12, biotin, and their interaction on the EB and feed efficiency (P > 0.05).

       Plasma Variables

      The effects of biotin and VB12 on plasma variables are listed in Table 3. When cows were injected with VB12, plasma nonesterified fatty acids concentration was lower throughout the entire experimental period compared with that of cows not treated with VB12 (P < 0.01). Reduced plasma BHB was observed in cows treated with VB12, compared with that of cows without VB12 injection (P = 0.04). The SOD concentration in the plasma was significantly greater in VB12-treated cows compared with that of animals not supplemented with VB12 (P = 0.05). Supplementation of both VB12 and biotin tended to have an interactive effect on improving plasma SOD concentration (P = 0.08). Plasma biotin was greater in cows given biotin compared with animals without biotin supplementation (P = 0.04). Plasma VB12 was greater in cows injected with VB12 compared with that of animals without VB12. Moreover, no significant effects of VB12 × week or biotin × week on plasma variables was observed (P > 0.05, data not shown).
      Table 3Effects of dietary biotin (Bio) and intramuscular injection of vitamin B12 (VB12) on plasma variables in prepartum and postpartum dairy cows
      Sampling times: the blood was sampled at −10, 0, 8, 15, 29, 43 and 57 d relative to calving.
      ItemTreatmentSEMP-value
      ControlBioVB12Bio + VB12BioVB12Biotin + VB12
      Glucose, mmol/L3.443.443.433.540.0370.320.330.33
      Cholesterol, mmol/L3.553.103.623.660.2240.360.160.27
      BUN, mmol/L4.584.414.744.810.1180.810.110.50
      Nonesterified fatty acids, μmol/L323.9303.8216.2246.315.040.82<0.010.24
      BHB, μmol/L550.5522.0471.1499.224.420.930.040.21
      SOD,
      SOD = superoxide dismutase.
      U/mL
      110.4105.9111.1114.51.600.820.040.08
      Biotin,
      Sampling time: the blood was sampled at 57 d relative to calving.
      ng/L
      7321,2587511,21422.50.040.950.89
      VB12,
      Sampling time: the blood was sampled at 57 d relative to calving.
      ng/L
      19516241844152.20.93<0.010.59
      1 Sampling times: the blood was sampled at −10, 0, 8, 15, 29, 43 and 57 d relative to calving.
      2 SOD = superoxide dismutase.
      3 Sampling time: the blood was sampled at 57 d relative to calving.

      DISCUSSION

      In the current study, DMI was approximately 1.7 kg/d higher in VB12-injected cows compared with cows without injection. This result is inconsistent with the studies of
      • Girard C.L.
      • Matte J.J.
      Effects of intramuscular injections of vitamin B12 on lactation performance of dairy cows fed dietary supplements of folic acid and rumen-protected methionine.
      and
      • Duplessis M.
      • Lapierre H.
      • Pellerin D.
      • Laforest J.P.
      • Girard C.L.
      Effects of intramuscular injections of folic acid, vitamin B12, or both, on lactational performance and energy status of multiparous dairy cows.
      , in which VB12 injection had no effects in DMI and milk production in dairy cows.
      • Girard C.L.
      • Matte J.J.
      Effects of intramuscular injections of vitamin B12 on lactation performance of dairy cows fed dietary supplements of folic acid and rumen-protected methionine.
      conducted their study in primiparous lactating cows (starting at 4 wk postpartum), different from our study starting at 3 wk prepartum with multiparous dairy cows. It was reported that when plasma VB12 concentration is greater than 200 pg/mL in multiparous cows, additional folates increased milk protein content, milk yield, and milk protein yield, but no improved performance was observed in primiparous cows whose plasma VB12 concentrations were lower (
      • Girard C.L.
      • Matte J.J.
      • Tremblay G.F.
      Gestation and lactation of dairy cows: A role for folic acid?.
      ;
      • Girard C.L.
      • Matte J.J.
      Dietary supplements of folic acid during lactation: Effects on the performance of dairy cows.
      ).
      • Akins M.S.
      • Bertics S.J.
      • Socha M.T.
      • Shaver R.D.
      Effects of cobalt supplementation and vitamin B12 injections on lactation performance and metabolism of Holstein dairy cows.
      observed no effect of VB12 supplementation in cows with a low folate status (lower than 20 ng/mL). In the current study, plasma VB12 concentration of animals in control group was lower than 200 pg/mL (about 170 pg/mL), whereas VB12 level was over 400 pg/mL after VB12 injection. Thus, it is possible that folate status of cows in the current study (not measured) was high enough to allow a detectable effect on DMI and milk yield. We inferred that effect of VB12 injection on DMI and lactation performance of dairy cows depended on parity and availability of VB12 and folate in the blood.
      Biotin supplementation (30 mg/d) did not affect DMI, which is in agreement with previous studies [20 mg/d for prepartum and 30 mg/d for postpartum in
      • Rosendo O.
      • Staples C.R.
      • Mcdowell L.R.
      • Mcmahon R.
      • Badinga L.
      • Martin F.G.
      • Shearer J.F.
      • Seymour W.M.
      • Wilkinson N.S.
      Effects of biotin supplementation on peripartum performance and metabolites of Holstein cows.
      ; 10 or 20 mg/d in
      • Chen B.
      • Wang C.
      • Liu J.X.
      Effects of dietary biotin supplementation on performance and hoof quality of Chinese Holstein dairy cows.
      ], but is not consistent with
      • Zimmerly C.A.
      • Weiss W.P.
      Effects of supplemental dietary biotin on performance of Holstein cows during early lactation.
      and
      • Chen B.
      • Wang C.
      • Liu J.X.
      Effects of dietary biotin supplementation on performance and hoof quality of Chinese Holstein dairy cows.
      , who observed higher milk yield after biotin supplementation.
      • Chen B.
      • Wang C.
      • Liu J.X.
      Effects of dietary biotin supplementation on performance and hoof quality of Chinese Holstein dairy cows.
      started their work at the early lactating period (DIM = 40), whereas our experiment began at precalving. Although they started their study at prepartum stage,
      • Zimmerly C.A.
      • Weiss W.P.
      Effects of supplemental dietary biotin on performance of Holstein cows during early lactation.
      included both primiparous and multiparous cows. It is not clear whether the response is different between the cows of different parities. Therefore, the different responses to biotin addition among different studies could be attributed to different lactation stages and parity of the cows. However, we observed that biotin addition increased protein (6 and 8 wk) and lactose yield (from 6 to 8 wk) in a time-dependent manner compared with cows without biotin. The improved lactose yield could be associated with the positive effect of biotin on hepatic gluconeogenesis in dairy cows (
      • Majee D.N.
      • Schwab E.C.
      • Bertics S.J.
      • Seymour W.M.
      • Shaver R.D.
      Lactation performance by dairy cows fed supplemental biotin and a B-vitamin blend.
      ) through increasing propionyl-CoA carboxylase decarboxylase activity (
      • Ouattara B.
      • Duplessis M.
      • Girard C.L.
      Optimization and validation of a reversed-phase high performance liquid chromatography method for the measurement of bovine liver methylmalonyl-coenzyme a mutase activity.
      ). Moreover, the enhanced hepatic gluconeogenesis is beneficial to reduce AA deamination in the liver and to increase AA utilization efficiency in the mammary gland (
      • Seymour W.M.
      • Polan C.E.
      • Herbein J.H.
      Effects of dietary protein degradability and casein or amino acid infusions on production and plasma amino acids in dairy cows.
      ), resulting in more AA flowing to mammary gland for milk protein synthesis. Under the experimental conditions of our study, the positive effects of a biotin supplementation on protein and lactose yields were visible only after 6 wk postpartum.
      An interaction on lactation performance between biotin and VB12 could be expected based on their respective roles in propionate utilization for gluconeogenesis (
      • Padmakumar R.
      • Padmakumar R.
      • Banerjee R.
      Evidence that cobalt-carbon bond homolysis is coupled to hydrogen atom abstraction from substrate in methylmalonyl-CoA mutase.
      ;
      • Scott J.M.
      Folate and vitamin B12.
      ;
      • Hügler M.
      • Krieger R.S.
      • Jahn M.
      • Fuchs G.
      Characterization of acetyl-CoA/propionyl-CoA carboxylase in Metallosphaera sedula. Carboxylating enzyme in the 3-hydroxypropionate cycle for autotrophic carbon fixation.
      ). However, under the conditions of our experiment, the biotin × VB12 interactions were not significant for plasma glucose concentration or milk lactose yield and lactation performance.
      Compared with animals without VB12 injection, the reduced BW loss, lower body fat mobilization (lower nonesterified fatty acids and BHB in the plasma) and lower oxidative stress in VB12-injected cows may partly attributed to the increased DMI (
      • Wang D.M.
      • Wang C.
      • Liu H.Y.
      • Liu J.X.
      • Ferguson J.D.
      Effects of rumen-protected γ-aminobutyric acid on feed intake, lactation performance, and antioxidative status in early lactating dairy cows.
      ;
      • Konvičná J.
      • Vargová M.
      • Paulíková I.
      • Kovcáč G.
      • Kostecká Z.
      Oxidative stress and antioxidant status in dairy cows during prepartal and postpartal periods.
      ;
      • Kuhla B.
      • Metges C.C.
      • Hammon H.M.
      Endogenous and dietary lipids influencing feed intake and energy metabolism of periparturient dairy cows.
      ). In contrast, biotin had limited effect on BW change due to its limited effect on DMI in the current study.

      CONCLUSIONS

      Weekly injection of VB12, starting at the prepartum stage, increased postpartum DMI and lactation performance and reduced BW loss of dairy cows. Biotin had a limited effect on overall lactation performance but did show a time-dependent effect on protein and lactose yield. Under the present experimental conditions, we found no interactive effect of a combined supplement of biotin and VB12 on lactation performance of dairy cows.

      ACKNOWLEDGMENTS

      This research was supported by the grants from the National Key Research and Development Program of China (2016YFD0500504) and the China Agriculture Research System (CARS-36, Ministry of Agriculture and Rural Affairs of China, Beijing). Authors gratefully thank all of the group members of the Hangzhou Zhengxing Animal Industries (Hangzhou, China) for their assistance in the feeding and care of the cows. We also acknowledge the members of the Institute of Dairy Science of Zhejiang University (Hangzhou, China) for their assistance in the field sampling and data analysis.

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