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Lactation Performance by Dairy Cows Fed Supplemental Biotin and a B-Vitamin Blend

D. N. Majee^*, E. C. Schwab^*, S. J. Bertics^*, W. M. Seymour and R. D. Shaver^*

^* Department of Dairy Science, University of Wisconsin, Madison 53706
Roche Vitamins Inc., Parsippany, NJ 07054

Corresponding author:
R. D. Shaver; email: rdshaver{at}facstaff.wisc.edu.

	ABSTRACT

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 
The objective of Trial 1 was to evaluate in dairy cows the effects of dietary supplementation with biotin and a B-vitamin blend on dry matter intake (DMI), milk yield, composition and component yields, total tract nutrient digestion, and plasma metabolites. Twenty-four multiparous Holstein cows averaging 46 ± 8 d in milk at trial initiation were randomly assigned to treatments in a replicated 4 x 4 Latin square design with 28 d periods. The four treatments were: 1) a control diet (C) with no supplemental B-vitamins; 2) C plus supplemental biotin at 20 mg/d (B); 3) C plus supplemental thiamin (150 mg/d), riboflavin (150 mg/d), pyridoxine (120 mg/d), B₁₂ (0.5 mg/d), niacin (3000 mg/d), pantothenic acid (475 mg/d), folic acid (100 mg/d), and biotin (20 mg/d) (BBVIT1X); 4) C plus supplemental thiamin (300 mg/d), riboflavin (300 mg/d), pyridoxine (240 mg/d), B₁₂ (1.0 mg/d), niacin (6000 mg/d), pantothenic acid (950 mg/d), folic acid (200 mg/d), and biotin (40 mg/d) (BBVIT2X). Intake of DM was increased 0.7 kg/d for B vs. C and BBVIT1X and 1.3 kg/d for B vs. BBVIT2X. Milk yield was increased 1.7 kg/d for B vs. C. For BBVIT1X, milk yield was similar to B and BBVIT2X and tended to be higher than C. Yields of milk protein and lactose but not fat were higher for B than C. For BBVIT1X, milk component yields were similar to B and tended to be higher than C, with the exception of lactose yield where BBVIT1X was higher than C. The objective of Trial 2 was to evaluate DMI and milk yield, composition and component yields by dairy cows fed diets supplemented with either 40 mg/d biotin or the B-vitamin blend (BBVIT1X) compared to cows supplemented with 20 mg/d dietary biotin. Neither the 40 mg/d biotin treatment nor the B-vitamin blend enhanced lactation performance over the 20 mg/d biotin treatment. Biotin efficacy in short-term trials suggests that biotin may improve milk yield directly via effects on intake and (or) nutrient metabolism rather than indirectly via improved hoof health. More research is needed to determine the mode of action for supplemental dietary biotin.

Key Words: biotin • B-vitamins • dairy cows • milk production

Abbreviation key: B = Biotin at 1X dose in Trial 1, B1X = Biotin at 1X dose in Trial 2, B2X = Biotin at 2X dose in Trial 2, BBVIT1X = Biotin and B-vitamin blend at 1X dose in Trials 1 and 2, BBVIT2X = Biotin and B-vitamin blend at 2X dose in Trial 1, C = Control in Trial 1

	INTRODUCTION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 
The Dairy NRC (2001) does not provide recommended B-vitamin allowances for lactating dairy cows because of a lack of research on B-vitamin requirements for gestation, health, and milk production. Also, it was felt that, in general, B-vitamin requirements could be met through synthesis by ruminal microorganisms and escape of basal dietary sources from the rumen.

However, recent evidence of improved lactation performance from B-vitamin supplementation can be found in the literature. Supplemental dietary thiamin (150 to 300 mg/d) increased milk and component yields by cows fed low NDF, high NSC diets (Shaver and Bal, 2000). Supplemental dietary folic acid (1.3 to 2.5 g/d) increased milk yield in multiparous but not primiparous cows (Girard and Matte, 1998). Girard and Matte (1997) reported a numerical increase (P > 0.10) in milk production in response to vitamin B₁₂ injections (10 mg/wk, i.m.) in primiparous cows. Because of extensive ruminal degradation of choline (Sharma and Erdman, 1988) and positive milk yield and fat test responses to choline supplemented in a ruminally-protected form (Erdman, 1994), ruminally-protected choline products are available commercially. Supplemental dietary biotin (20 mg/d) improved hoof health and DHI-estimated milk yield (Midla et al., 1998). Milk yield increased linearly with 0, 10, and 20 mg/d biotin supplemented in diets from 14 d prepartum through 100 d postpartum (Zimmerly and Weiss, 2001). Although niacin is a widely used supplemental B-vitamin by the feed industry, the Dairy NRC (2001) concluded from a summary of 25 trials that the routine use of niacin to enhance lactation performance by dairy cows or to minimize the risk of ketosis and fatty liver is not recommended.

The objective of Trial 1 was to evaluate in dairy cows the effects of dietary supplementation with biotin and a B-vitamin blend on DMI, milk yield, composition and component yields, total tract nutrient digestion, and plasma metabolites. The objective of Trial 2 was to evaluate DMI and milk yield, composition and component yields by dairy cows fed diets supplemented with 40 mg/d biotin or the B-vitamin blend compared to cows fed the 20 mg/d biotin dose tested in Trial 1.

	MATERIALS AND METHODS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 
Trial 1
The Research Animal and Resource Center of the College of Agriculture and Life Sciences at the University of Wisconsin-Madison approved the experimental protocol. Twenty-four multiparous Holstein cows were randomly assigned to treatments in a replicated 4 x 4 Latin square design with 28 d periods. Cows were housed and fed in tie-stalls. The first 14 d of each period were for diet adaptation and sampling occurred on d 15 through 28. Cows averaged 46 ± 8 DIM at trial initiation and were injected with bovine somatotropin (Posilac, Monsanto Company, St. Louis, MO) every 14 d starting on d 1 of the experiment. The four treatments were: 1) control diet (C) with no supplemental B-vitamins; 2) C plus supplemental biotin at 20 mg/d (Rovimix H-2 spray-dried 2%) (B); 3) C plus supplemental thiamin (150 mg/d; Rovimix B₁ fine-crystalline powder 92%), riboflavin (150 mg/d; Rovimix B₂ spray-dried 80%), pyridoxine (120 mg/d; Rovimix B₆ fine-crystalline powder 82%), B₁₂ (0.5 mg/d; B₁₂ crystalline powder 1% dilution), niacin (3000 mg/d; Rovimix niacin fine-crystalline powder 99.5%), pantothenic acid (475 mg/d; Rovimix Calpan spray-dried 92%), folic acid (100 mg/d; Rovimix Folic spray-dried 80%), and biotin (20 mg/d; Rovimix H-2 spray-dried 2%) (BBVIT1X); 4) C plus supplemental thiamin (300 mg/d; Rovimix B₁ fine-crystalline powder 92%), riboflavin (300 mg/d; Rovimix B₂ spray-dried 80%), pyridoxine (240 mg/d; Rovimix B₆ fine-crystalline powder 82%), B₁₂ (1.0 mg/d; B₁₂ crystalline powder 1% dilution), niacin (6000 mg/d; Rovimix Niacin fine-crystalline powder 99.5%), pantothenic acid (950 mg/d; Rovimix Calpan spray-dried 92%), folic acid (200 mg/d; Rovimix Folic spray-dried 80%), and biotin (40 mg/d; Rovimix H-2 spray-dried 2%) (BBVIT2X). Average daily intakes of supplemental B-vitamins calculated from premix formulations and actual DMI are presented in Table 1. Control and treatment premixes were included in the respective TMR at 0.46% of DM for average daily premix intakes of 115 g/d. The four mo supply of premixes needed for the trial were prepared immediately prior to the start of the trial by Roche Vitamins, Inc. (Parsippany, NJ) at their Blend Plant in Ames, IA. Shurson et al. (1996) reported average monthly losses of B-vitamin potency after four mo in storage ranging from 0 to 8% across the B-vitamins in a vitamin premix containing choline chloride, which increases the breakdown of B-vitamins; choline chloride was not a premix ingredient in our study. Midla et al. (1998) found no loss of B-vitamin potency 24 h after mixing in a TMR. The formulation target levels for biotin in treatment B and B₁₂, biotin, folic acid, and niacin in BBVIT1X were obtained from Roche guidelines for ruminants (Roche, 2000). The formulation target level for thiamin in BBVIT1X was based on Shaver and Bal (2000). The formulation target levels for pantothenic acid, pyridoxine, and riboflavin in BBVIT1X were based on Roche guidelines for lactating sows (NRC, 1988) extrapolated to lactating dairy cows according to BW and feed intake differences (Roche, 2000). The lower end of the range of Roche B-vitamin guidelines for ruminants and sows (Roche, 2000) was used to develop formulation target levels for BBVIT1X, because of cost considerations for the B-vitamin blend. The BBVIT2X treatment was included to allow for a dose-response comparison.

Trial 2
The conduct of Trial 2 was similar to that of Trial 1 and only the differences between the two trials are presented. Twenty-four multiparous Holstein cows (84 ± 15 DIM at trial initiation) were randomly assigned to treatments in a replicated 3 x 3 Latin square design with 28 d periods. The three treatments were: 1) diet formulated to provide supplemental biotin at 20 mg/hd/d (Rovimix H-2 spray-dried 2%) (B1X); 2) same diet as B1X formulated to provide supplemental biotin at 40 mg/hd/d (Rovimix H-2 spray-dried 2%) (B2X); 3) same diet as B1X formulated to provide supplemental thiamin (150 mg/d; Rovimix B₁ fine-crystalline powder 92%), riboflavin (150 mg/d; Rovimix B₂ spray-dried 80%), pyridoxine (120 mg/d; Rovimix B₆ fine-crystalline powder 82%), B₁₂ (0.5 mg/d; B₁₂ crystalline powder 1% dilution), niacin (3000 mg/d; Rovimix niacin fine-crystalline powder 99.5%), pantothenic acid (475 mg/d; Rovimix Calpan spray-dried 92%), folic acid (100 mg/d; Rovimix Folic spray-dried 80%), and biotin (20 mg/d; Rovimix H-2 spray-dried 2%) (BBVIT1X). Average daily intakes of supplemental B-vitamins calculated from premix formulations and actual DMI are presented in Table 1. The TMR composition is presented in Table 2.

	RESULTS AND DISCUSSION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 
Dietary nutrient composition and lactation performance data for both trials are presented in Tables 2 and 3, respectively. Digestibility and plasma data for Trial 1 are presented in Table 4.

Milk yield was increased (P < 0.05) 1.7 kg/d for B vs. C. It cannot be determined from this study whether the increase in milk yield was in response to the higher DMI or vice versa. Zimmerly and Weiss (2001) reported that milk yield increased 0.9 kg/d and 2.8 kg/d linearly above control (no supplemental biotin) for 10 and 20 mg/d supplemental dietary biotin, respectively, during the first 100 DIM with no statistical change in DMI. Margerison et al. (2002) reported that milk yield was increased 2.0 kg/d above control (no supplemental biotin) by 20 mg/d supplemental dietary biotin during the first 120 DIM. For BBVIT1X, milk yield was similar (P > 0.10) to B and BBVIT2X (38.3 vs. 38.9 and 37.5 kg/d, respectively) and tended (P < 0.10) to be higher than C (38.3 vs. 37.2 kg/d). Milk yield was similar (P > 0.10) for C and BBVIT2X. Feed efficiency (kg milk/kg DMI) was similar (P > 0.10) across treatments and averaged 1.53 (data not presented in table).

Yields of milk protein and lactose but not fat were higher (P < 0.05) for B than C. Zimmerly and Weiss (2001) reported that yield of milk protein but not fat was increased for biotin-supplemented vs. unsupplemented cows. For BBVIT1X, milk component yields were similar (P > 0.10) to B and tended (P < 0.10) to be higher than C, with the exception of lactose yield where BBVIT1X was higher than C (P < 0.05). Yields of milk fat and lactose but not protein were lower (P < 0.05) for BBVIT2X than BBVIT1X. Milk composition was not different across treatments (P > 0.10) except for lactose percentage where B and BBVIT1X were higher (P < 0.05) than C and BBVIT2X. Milk fat and protein percentages were unaffected by dietary biotin supplementation in the studies of Margerison et al. (2002) and Zimmerly and Weiss (2001). In the present experiment lactose yield of cows supplemented with 20 mg/d dietary biotin was increased by 110 g/d compared to the control. This increase in lactose yield is equivalent to 220 g/d of glucose. Biotin is a required co-factor for enzymes involved in propionate utilization and gluconeogenesis (McDowell, 2000). Supplemental biotin may have alleviated a limitation on the activities of these enzymes leading to increased lactose synthesis. We can provide no clear explanation for reduced lactation performance of cows fed BBVIT2X compared to cows fed B or BBVIT1X, which may have been related to lower DMI for BBVIT2X (24.4 vs. 25.0 to 25.7 kg/d). It is unclear why DMI was lower for BBVIT2X, but poor palatability and (or) negative interactions among the B-vitamins at the high dosages may have been factors.

Apparent total-tract DM, OM, and NDF digestibilities were not different across treatments (P > 0.10). This suggests that the observed intake differences were not related to changes in diet digestibility. Our data do not support the suggestion of Zimmerly and Weiss (2001) that biotin increases milk yield partially through improved fiber digestibility. Dietary biotin supplementation had no effect on ruminal VFA molar percentages in the trial of Zimmerly and Weiss (2001). Zimmerly and Weiss (2001) suggested that biotin increases milk yield partially through increased glucose production, because biotin is a cofactor for the glucogenic enzyme pyruvate carboxylase and for the enzyme propionyl-CoA carboxylase (McDowell, 2000). Plasma glucose, NEFA and BHBA concentrations were not different across treatments (P > 0.10). Similarly, dietary biotin supplementation had no effect on plasma glucose and NEFA concentrations measured at 1, 30, 60, and 100 DIM in the trial of Zimmerly and Weiss (2001). Plasma metabolite concentrations do not measure the amount of metabolite produced or utilized, and therefore may not be good indicators of treatment effects on nutrient metabolism.

Biotin efficacy in the short-term (100 d) continuous lactation trial of Zimmerly and Weiss (2001) and our short-term (28 d periods) Latin square trial suggests that biotin may improve milk yield directly via effects on intake, digestion, and (or) metabolism rather than only indirectly via improved hoof health (Midla, 1998). However, no measurement of hoof health was made in the present experiment. Zinn et al. (1987) measured duodenal flows of several B-vitamins, and reported extensive ruminal microbial degradation of supplemental B-vitamins with the exception of biotin and B₆, which largely escaped ruminal degradation. Their observations regarding microbial degradation may partially explain the variable responses to supplemental ruminally-unprotected B-vitamins reported in the literature including the lack of response to BBVIT1X over B in this study, and the more consistent positive efficacy reports for supplemental ruminally-unprotected biotin including the response to B over C in this study. Richards et al. (2002) reported that milk yield was increased 3.7 kg/d above control (no supplemental B-vitamins) by a B-vitamin blend (rumen protected choline, niacin, B₁₂, biotin, folic acid, and thiamin) during the first 90 DIM, but the composition of the blend was not provided because a proprietary product was being evaluated. Stage of lactation may have affected the response to B-vitamins in our trial. Extensive microbial degradation of some supplemental B-vitamins suggests a possible role for ruminally-protected B-vitamins to improve efficacy, consistency of response, and (or) reduce minimum effective dosages of B-vitamin blends, but more research on duodenal flows and animal requirements for the various B-vitamins is needed to improve supplement development. Based on positive efficacy reported for biotin in our study and the studies of Margerison et al. (2002), Midla et al. (1998), and Zimmerly and Weiss (2001), it does not appear that biotin is a B-vitamin that requires ruminal protection.

Trial 2
Body weight, DMI, and milk yield were not different (P > 0.10) among the treatments and averaged 674, 25.8, and 41.0 kg/d, respectively. Feed efficiency (kg milk/kg DMI) was similar (P > 0.10) across treatments and averaged 1.61 (data not presented in table). Milk composition and component yields were not different (P > 0.10) among the treatments. The 40 mg/d biotin treatment (B2X) or the B-vitamin blend (BBVIT1X) did not enhance lactation performance over the 20 mg/d biotin treatment (B1X). Although, Kluenter et al. (1993) reported linear increases in plasma and milk biotin concentrations with increasing incremental intakes of oral supplemental biotin from 0 to 80 mg/d.

	CONCLUSIONS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 
In Trial 1, DMI and milk yield were 0.7 and 1.7 kg/d, respectively, higher for cows supplemented with 20 mg/d dietary biotin compared to control cows. There was no further benefit to supplementing a B-vitamin blend along with biotin. In Trial 2, there was no benefit to dietary supplementation with 40 mg/d biotin or the B-vitamin blend over 20 mg/d supplemental dietary biotin. Conclusions regarding the B-vitamin blend should not be interpreted too broadly, because duodenal flows and animal requirements for the various B-vitamins are not well defined and a blend with a different B-vitamin composition or fed earlier in lactation may elicit a different response. Income minus feed cost is expected to increase $0.20/d per cow from dietary biotin supplementation assuming a 1.7 kg/d increase in milk yield at a milk price of $0.26/kg, $0.15/d cost for 20 mg/d supplemental biotin, and a 0.7 kg/d increase in DMI at a ration cost of $0.15/kg DM. More research is needed to determine the mode of action for supplemental dietary biotin.

Received for publication August 17, 2002. Accepted for publication February 2, 2003.

	REFERENCES

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSSION CONCLUSIONS REFERENCES

 


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