Dietary Choline for the Lactating Cow:

Possible Effects on Milk Fat Synthesis ~

R. A. E R D M A N , R. D. SHAVER, 2 and J. H. V A N D E R S A L L
Department of Animal Sciences

University of Maryland
College Park 20742

ABSTRACT

ment for dietary choline demonstrates that the

preweaning calf lacks the ability to synthesize
sufficient choline to maintain normal b o d y
functions. Water soluble vitamins are thought
to be synthesized in sufficient quantities by
rumen microorganisms to supply the animal's
dietary needs (16, 20, 21, 22, 23). However, in
most studies with rumen microbial synthesis of
vitamins, actual amounts of water soluble vitamins presented to the duodenum never were
measured, and rates of synthesis were determined only by change of vitamin concentration
between feed and rumen digesta (20, 21, 22,
23). In none of these experiments was ruminal
choline synthesis studied.

Two experiments with low forage

diets tested effects of added dietary choline on milk fat synthesis. In Experiment
1, 18 Holstein cows were in a 12-wk trial.
Choline (0, 1.5, and 3 g/kg) was added to
the concentrate portion of a 70% concentrate, 30% corn silage diet (dry matter).
Treatments were applied in a switchback
design with 4-wk experimental periods.
Highest choline increased milk fat b y .34
percentage units and fat-corrected milk
by 2 kg per day over controls. Rumen pH
and volatile fatty acids were similar
among treatments. Free fatty acids of
blood serum were reduced 3.0 rag/100
ml by 3 g/kg choline. In Experiment 2
with four mature Holsteins, choline
was added (0, 2, 4, and 6 g/kg concentrate) in 25% corn silage and 75% concentrate diets. Treatments were applied in a
4 4 Latin square design. Addition of 4
g/kg choline resulted in 3.7 kg/day and
.77 percentage unit increases of fatcorrected milk and milk fat percent,
respectively, whereas fat-corrected milk
and milk fat percent at 6 g/kg were only
slightly higher than controls. We hypothesized that choline aided transport of
mobilized free fatty acids from adipose
tissue through the liver to the mammary
gland.

Work with the use of added dietary choline

in ruminant rations is little. Dyer et al. (8) reported increased average daily gain of growing
cattle from addition of choline up to 1.3 g/kg
dry matter consumed. Other reports with
finishing cattle reported slight increases (28) of
weight gains, whereas in some experiments (13,
30) no response occurred after addition of
dietary choline.

INTRODUCTION

Choline is an essential dietary nutrient in

preweanling ruminants (15, 25). The requireJanuary 31, 1983.
1Scientific Article No. A-3360, Contribution No.
6432 of the Maryland Agricultural Experiment Station.
2Department of Dairy Science, University of Wisconsin, Madison 53706.
Received

1984 J Dairy Sci 67:410-415

Workers (19) suggested possible roles of

methionine and other lipotropic agents in ruminant lipid metabolism. Addition of dietary
methionine slightly increased milk fat percent
(2, 6). However, the mechanism for the response to methionine is unknown. In rats
choline regulated synthesis of lipoproteins in
the liver, and choline deficiency blocked synthesis of either the apolipoprotein or attachment of synthesized triglyceride to the
apolipoprotein in the liver (17, 24). If the cow
relies primarily on adipose tissue as a source of
long chain fatty acids for milk fat synthesis
(9, 27), then choline deficiency could block the
lipid transport mechanism to the mammary
gland. Feeding experiments were initiated to
test effects of added dietary choline on milk fat
synthesis.

410

TEC HNICAL NOTE

EXPERIMENTAL PROCEDURES
Experiment 1

Eighteen Holstein cows at least 60 days postp a r t u m were in the trial. Cows were fed individually t o t a l m i x e d diets containing 30% corn
silage and 70% c o n c e n t r a t e (dry matter) for a
2-wk a d j u s t m e n t prior to a 12-wk e x p e r i m e n t a l
period (Table 1). At the beginning o f the experimental period, cows were assigned rand o m l y to one o f three treatments: 1) control,
2) control plus 1.5 g choline, or 3) control plus
3 g choline (per kilogram c o n c e n t r a t e as fed).
Diets were applied in a switchback deSign (18)
with 4-wk periods. Diets were slightly deficient
in crude protein (25) because of responses to
m e t h i o n i n e h y d r o x y analog in low protein diets
(6). Low forage diets were fed to test effects of
dietary choline w h e n milk fat percent was
depressed.
Measurements included daily feed intake and
milk p r o d u c t i o n . Milk samples f r o m a.m. and
p.m. milkings were taken twice weekly.
Samples were c o m p o s i t e d b y weight and analyzed for milk fat and protein by Babcock and
dye binding procedures. B o d y weights were
measured 1 day each week. R u m e n samples
were taken b y stomach t u b e on the last day of

411

the e x p e r i m e n t at 4 h after feeding, and pH was

measured i m m e d i a t e l y . Samples t h e n were frozen at - 1 0 C and analyzed for volatile f a t t y
acids ( V F A ) as in (10). Feed samples for dry
m a t t e r (DM) analysis were taken weekly, and
silage and concentrates were analyzed for DM
by t o l u e n e distillation and oven drying at
100C. Acid-detergent fiber c o n t e n t of feeds
was analyzed by m e t h o d s o f Goering and Van
Soest (12). Choline c o n t e n t of diets was determined on c o m p o s i t e samples of c o n c e n t r a t e
and corn silage (15). Statistical analysis was by
the general linear m o d e l p r o c e d u r e (1) where
the regression m o d e l included period, treatment, and cow effects.
T r e a t m e n t effects, f r o m data f r o m the last 2
wk of each e x p e r i m e n t a l period, were separated
into linear and quadratic o r t h o g o n a l contrasts.
R u m e n data were analyzed b y analysis of variance for a c o m p l e t e l y r a n d o m i z e d design (7).
All data e x c e p t r u m e n m e a s u r e m e n t s are least
squares means.
Experiment 2

A second trial was initiated with m o r e choline in similar types of diets. F o u r m a t u r e Holstein cows were in the 12-wk e x p e r i m e n t . Cows
were fed individually diets containing 25% corn

TABLE 1. Composition of concentrates (% as fed).

Added choline, g/kg concentrate
Experiment 1

Experiment 2

1.5

3.0

Ground corn
Soybean meal (49%)
Corn gluten feed
Brewer's dried grains
Corn gluten meal
Trace mineralized salt
Dicalcium phosphate
Limestone
Potassium-magnesium sulfate
Potassium chloride
Vitamin A and D
Choline chloride (50/0)a

52.3
6.0
18.6
18.6
...
.7
1.3
1.0
.6
.6
.3
0

52.0
6.0
18.6
18.6

51.7
6.0
18.6
18.6

.7
1.3
1.0
.6
.6

.7
1.3
1.0
.6
.6

.3
.3

.3
.6

Total ration chemical composition

(dry matter basis)
Crude protein, %
Acid detergent fiber, %
Choline, g/kg

14.7
17.8
.74

14.2
17.9
1.74

14.2
17.9
2.52

Ingredient

84.5
84.1
83.7
. . . . . . . . . . . .
. . . . . . . . . . . .
.

il.;

83.3

ii.'o

ii.;

11.0

.6
1.1
.8
.7
1.1

.6
1.1
.8
.7
1.1

.6
1.1
.8
.7
1.1

.6
1.1
.8
.7
1.1

.2

.2
.4

.2
.8

.2
1.2

13.3
11.8
.38

13.4
11.9
1.90

13.5
12.0
3.58

13.6
12.0
5.38

asyntex Agribusiness, Inc., SSS, Springfield, MO 65805.

Journal of Dairy Science Vol. 67, No. 2, 1984

412

E R D M A N ET AL.

silage and 75% concentrate on a DM basis

(Table 1). Treatments included 0, 2, 4, and 6
g/kg choline added to the concentrate portion
of the diet. Treatments were applied in a 4 4
Latin square design with 3-wk experimental
periods.
Measurements included daily feed intake and
milk production. Cows were weighed weekly.
Milk was sampled for a.m. and p.m. twice
weekly for milk fat determinations Feeds were
sampled weekly for DM determination and
chemical analyses as in Experiment 1. Data
from the last week of each experimental period
were analyzed by analysis of variance (7).
Treatment comparisons were with orthogonal
contrasts for linear, quadratic, and cubic
effects, and individual mean differences were
determined b y Fisher's least significant difference test only after a significant F-test (7).

OX

AND

O0

v-~

~:

xO~'..,t~.'~-

.ooxo

:,4

~
RESULTS

'

" :

DISCUSSION

Intake and production data for both experiments are in Table 2. Choline had no effect
( P > . I ) on total DM intake or intake as a percentage of b o d y weight in either trial. Intake of
DM was less in Experiment 2 (13.8 kg/day)
than Experiment 1 (19.8 kg/day); reasons for
this are not clear because cows in Experiment 2
were producing approximately 25 kg per day
prior to the trial. Others (7, 27, 29) have reported no intake effects of up to 1.3 g/kg DM
added choline in beef cattle diets.
Choline had no effect ( P > . I ) on milk production in either trial. Milk fat percentage increased from 3.43 to 3.77% by increasing added
choline from 0 to 3 g/kg concentrate in Experiment 1, although differences only approached
significance at 10% probability. In Experiment
2, choline increased ( P < . I ) milk fat .77 percentage units, where 4 g/kg were added whereas
2 and 6 g/kg added choline did not alter milk
fat percentage. Numerical increases of milk fat
percent in Experiment 2 were greater than in
Experiment 1, and amounts of added choline
were also greater. Milk protein percent was unaffected ( P > . I ) by treatment in Experiment 1
(Table 2) and for that reason was not examined
in the second experiment.
Fat-corrected milk production increased 2
kg/day by addition of 3 g/day choline in
Experiment 1, although differences only approached significance at 10%. Added choline
J o u r n a l o f D a i r y S c i e n c e Vol. 67, No. 2, 1 9 8 4

0
,'d

'

'

.-4

.~

t~

m_

.~

'

'

.~
=

"7.
o

~8
~a

e4
.~
~:
~-

~= ~=~ ~-~ ~ . ~

,-4

TECHNICAL NOTE
up to 4 g/kg concentrate resulted in 3.7 kg increase ( P < . I ) of fat-corrected milk production
in Experiment 2 (Table 2). Trends were similar
for fat yield in both Experiments 1 and 2
(P<.I). Milk fat percents for control animals
were 3.43 and 2.64 in Experiments 1 and 2.
Although percent corn silage was almost identical (30 vs. 25%), milk fat percent was probably not depressed in Experiment 1 because acid
detergent fiber percentage of total ration was
much higher (17.8 vs. 11.8) because of more
acid detergent fiber in corn silage. It is possible
the response to comparable amounts of added
choline (Table 2) may be similar regardless of
whether or not cows are fat depressed.
Rumen pH and V F A patterns were not affected by choline addition in Experiment 1
(Table 3). These data represent samples taken
only in the last experimental period. Triglycerides in blood serum were not affected
( P > . I ) by addition of choline, although there
was a slight increase by the 3 g/kg concentrate
treatment. Addition of 3 g/kg choline reduced
(P<.01) serum fatty acids 3.0 and 5.5 rag/100
ml compared to control and 1.5 g/kg treatment
groups. Blood samples in Experiment 2 were
lost. Blood free fatty acid data from Experiment 1 suggested that choline may have an
effect on transport and mobilization of free
fatty acids from adipose tissue.

413
DISCUSSION

The possible role of choline and other tipotropic agents in cows with ketosis and fatty
liver was suggested (11, 19), Data from our
experiment may suggest a possible role of choline in milk fat depression. Recent work with
fasting-induced
and
spontaneous
ketosis
demonstrates the effect of mobilization of fatty
acids from adipose on liver fat accumulation (4,
11). These studies suggested an inability of the
liver to release synthesized triglycerides resulting from fatty acids taken up b y the liver in the
ketotic lactating cow.
Decreasing free fatty acids in Experiment 1
suggest that choline may be affecting transport
of free fatty acids, and this could be explained
by increased transport of synthesized triglycerides from the liver. Impaired release could be
caused b y either a lack of adequate apolipoprorein synthesis or a blockage of attachment of
triglycerides to apolipoproteins which blocks
release from the liver (17, 24). Because choline
may play a critical role in both steps (17, 24),
it may be that a choline deficiency plays a critical role in transport of triglycerides from the
liver in lactating cows.
Inadequate dietary choline in milk fat
depression may be related to diet. Increasing
proportions of concentrate in the ration associated with milk fat depression would lower in-

TABLE 3. Effect of added choline on rumen and blood measurements (Experiment 1).

Added choline, g/kg concentrate

1.5

3.0

6
5.89

6
6.11

6
6.01

".16--

Molar %
Acetate
Prop innate
Butyrate
Acetate/propionate

52.7
36.7
10.6
1.43

53.6
37.0
9.4
1.45

54.7
35.0
10.3
1.47

2.7
3.2
1.2
.25

Serum, mg/100 ml
Triglyceride
Free fatty acids

14.4
7.6 ab

14.6
10.1 b

18.9
4.6 a

2.2
1.2Q

Item
Observations
Rumen pH

SE

a'bMeans in the same row with different superscripts differ (P<.01).

Qsignificant quadratic effect (P<.01).
Journal of Dairy Science Vol. 67, No. 2, 1984

414

ERDMAN ET AL.

take o f d i e t a r y c h o l i n e b e c a u s e cereal grains are

t y p i c a l l y low in c h o l i n e c o n t e n t . C h o l i n e d o e s
n o t a p p e a r t o have a n y e f f e c t o n r u m e n pH o r
m o l a r p r o p o r t i o n s of a c e t a t e a n d p r o p i o n a t e
b a s e d o n d a t a f r o m E x p e r i m e n t 1 a n d (29).
This suggests t h a t t h e e f f e c t o f a d d e d c h o l i n e ,
if any, o n m i l k fat p r o d u c t i o n is n o t r e l a t e d t o
changes o f r u m e n f e r m e n t a t i o n .
R u m e n m e t a b o l i s m o f d i e t a r y c h o l i n e in t h e
f o r m of p l a n t p h o s p h o l i p i d is relatively u n k n o w n . However, w o r k b y Neill et al. (26)
d e m o n s t r a t e d t h a t 14C labeled p h o s p h a t i d y l
c h o l i n e was d e g r a d e d r a p i d l y t o t r i m e t h y l a m i n e
in t h e r u m e n . Diets in t h e s e studies (26) were
p r i m a r i l y forage a n d c o n c e n t r a t i o n of a d d e d
c h o l i n e was 100 t o 2 0 0 t i m e s less t h a n in o u r
s t u d y . Thus, b y p a s s o f d i e t a r y c h o l i n e is possible if c h o l i n e d e g r a d a t i o n s y s t e m s in t h e
r u m e n can b e o v e r w h e l m e d at high supplementation.
A t least one species o f r u m e n p r o t o z o a ,
Entodiniurn caudatum, has a g r o w t h requirem e n t f o r c h o l i n e (3). W o r k w i t h pure c u l t u r e s
of t h e s e species d e m o n s t r a t e d t h a t free c h o l i n e
is a b s o r b e d rapidly w h e n a d d e d to c u l t u r e
m e d i u m (5). L i m i t e d w o r k w i t h f l o w o f c h o l i n e
to t h e d u o d e n u m in s h e e p s h o w e d t h a t p r o t o zoa m a y b e t h e o n l y carriers o f c h o l i n e to t h e
l o w e r g a s t r o i n t e s t i n a l t r a c t (14). Because protozoal populatons tend to decline when rumen
pH decreases, it is possible t h a t f l o w o f c h o l i n e
t o t h e d u o d e n u m for a b s o r p t i o n is also red u c e d . H o w e v e r , it is u n k n o w n w h e t h e r c h o l i n e
is a b s o r b e d d i r e c t l y t h r o u g h t h e r u m e n wall.
R u m e n c o n d i t i o n s t h a t cause a decline in m i l k
f a t p e r c e n t , such as r e d u c e d p H a n d shifts in
a c e t a t e : p r o p i o n a t e ratios, m a y also b e associated w i t h a decline o f c h o l i n e flow if p r o t o zoa are t h e p r i m a r y carriers. With r e d u c e d
c h o l i n e , t r a n s p o r t o f triglycerides f r o m t h e liver
m a y b e b l o c k e d just as f o r k e t o t i c cows (4).
T h e e f f e c t of a d d e d c h o l i n e m a y b e m o r e pron o u n c e d f o r cows in early l a c t a t i o n w h e r e lipid
mobilization makes up a higher proportion of
t h e energy n e e d s o f t h e cow. T h e e f f i c i e n c y o f
a d d e d c h o l i n e in p r e v e n t i o n o f m i l k f a t depression is u n c e r t a i n a n d is o n l y suggested f r o m
data in these e x p e r i m e n t s . To d e t e r m i n e
w h e t h e r or n o t this t h e o r y is c o r r e c t will
d e m a n d m o r e e x p e r i m e n t a t i o n w i t h diets cont a i n i n g a w i d e r variety o f feed i n g r e d i e n t s a n d
cows a t h i g h e r p r o d u c t i o n .

Journal of Dairy Science Vol. 67, No. 2, 1984

ACKNOWLEDGMENTS

T h e a u t h o r s wish to a c k n o w l e d g e S y n t e x
Agribusiness, Inc., f o r c h o l i n e analysis a n d partial s u p p o r t of these e x p e r i m e n t s . We also
thank Jordan Thomas and Lauren McDermott
f o r care a n d assistance w i t h e x p e r i m e n t a l
animals a n d Gloria Switalski f o r l a b o r a t o r y
assistance.
REFERENCES

1 Barr, A. J., J. H. Goodnight, J. D. Sall, and J. T.

Helwig. 1976. A users guide to SAS 76. SAS Inst.,
Raleigh, NC.
2 Bhargava, P. K., D. E. Otterby, J. M. Murphy, and
J. D. Donker. 1977. Methionine hydroxy analog
for lactating cows J. Dairy Sei. 60:1594.
3 Brood, T. E., and R.M.C. Dawson. 1976. Role of
choline in the nutrition of the rumen protozoan
Entodiniurn caudaturn. J. Gen. Microbiol. 92:
391.
4 Brumby, P. E., M. Anderson, B. Tuckley, and J. E.
Storry. 1975. Lipid metabolism in the cow during
starvation-induced ketosis. Biochem. J. 146:609.
5 Bygrave, F. L., and R.M.C. Dawson. 1976. Phosphatidyl choline biosynthesis and choline transport
in the anaerobic protozoan Entodiniurn caudaturn.
Biochem. J. 160:481.
6 Chandler, P. T., C. A. Brown, R. P. Johnston, Jr.,
G. K. McCleod, R. D. McCartley, B. R. Moss, A. H.
Rakes, and L. D. Satter. 1976. Protein and
methionine hydroxy analog for lactating cows. J.
Dairy Sci. 59:1897.
7 Cochran, W. G., and B. M. Cox. 1957. Experimental design. John Wiley & Sons, New York, NY.
8 Dyer, J. A. 1969. Choline for fattening cattle.
Anita. Nutr. Health. October.
9 Emery, R. S. 1979. Deposition, secretion, transport and oxidation of fat in ruminants. J. Anim.
Sci. 48:1530.
10 Erdman, R. A., R. L. Botts, R. W. Hemken, and
L. S. Bull. 1980. Effects of sodium bicarbonate
and magnesium oxide on production and physiology in early lactation. J. Dairy Sci. 63:923.
11 Fronk, T. J., L. H. Schultz, and A. R. Hardie.
1980. Effect of dry period over conditioning on
subsequent metabolic disorders and performance
of dairy cows J. Dairy Sci. 63:1080.
12 Goering, H. K., and P. J. Van Soest. 1970. Forage
fiber analysis. Agric. Handbook No. 379. Agric.
Res. Ser., USDA, Washington, DC.
13 Harris, R. R., H. F. Yates, and J. E. Barrett, Jr.
1966. Choline in a roughage steer fattening ration.
J. Anim. Sci. 25:248.
14 John, A., and M. J. Wyatt. 1979. Phosphatidyl choline as a marker of duodenal flow of rumen protozoa in sheep. Proc. Nutr. Soc. 38:144A.
15 Johnson, B. C., H. H. Mitchell, and J. A. Pinkos.
1951. Choline deficiency in the calf. J. Nutr. 43:
379.
16 Lardinois, C. C., R. C. Mills, C. A. Elvehjen, and
E. B. Hart. 1941. Rumen synthesis of Vitamin B

T E C H N I C A L NOTE

17
18
19

20

21

22

23

complex as influenced by ration composition. J.

Dairy Sci. 27:579.
Lombardi, B. 1971. Effects of choline deficiency
on rat hepatocytes. Fed. Proc. 30:139.
Lucas, H. L. 1956. Switchback trials for m o r e than
two treamtents. J. Dairy Sci. 39:146.
McCarthy, R. D., G. A. Porter, and L. C. Griel, Jr.
1968. Bovine ketosis and depressed fat test in
milk. A problem in m e t h i o n i n e metabolism and
serum lipoprotein aberration. J. Dairy Sci. 51:
459.
McElroy, L. W., and H. Goss. 1941b. A quantitative s t u d y of the vitamins in the r u m e n c o n t e n t
of sheep and cows fed vitamin-low diets. IV.
Pantothenic acid. J. Nutr. 21:405.
McElroy, L. W., and H. Goss. 1941a. A quantitative s t u d y o f t h e vitamins in the r u m e n c o n t e n t of
sheep and cows fed vitamin-low diets. III. Thiamin.
J. Nutr. 21:163.
McElroy, L. W., and H. Goss. 1940. A quantitative s t u d y o f the vitamins in the r u m e n c o n t e n t s
of sheep and cows fed vitamin-low diets. I. Riboflavin and vitamin K. J. Nutr. 20: 527.
McElroy, L. W., and H. Goss. 1940. A quantitative s t u d y o f the vitamins in the r u m e n c o n t e n t of

24
25

26

27

28

29

30

415

sheep and cows fed vitamin-low d i e t s II. B 6 (Pyridine). J. Nutr. 20: 541.
Mooheryea, S. 1971. Action of choline in lipoprotein metabolism. Fed. Proc. 30:143.
National Research Council. 1978. Nutrient requirem e n t s o f dairy cattle. Natl. Acad. Sci., Washington, DC.
Neill, A., R. Derek, W. Grine, and R.M.C. Dawson.
1978. Conversion of choline m e t h y l groups
through tirmethylamine into m e t h a n e in the
rumen. Biochem. J. 170:529.
Patton, S., and R. G. Jensen. 1975. Lipid m e t a b o lism and m e m b r a n e functions of the m a m m a r y
gland. Prog. Chem. Fats Other Lipids 14:167.
Rumsey, T. S., and R. R. Oltjen. 1975. Sulfur and
choline in all concentrate beef finishing diets. J.
Anita. Sci. 41:416.
Swingle, R. S., and I. A. Dyer. 1970. E f f e c t s of
choline on r u m e n microbial metabolism. J. Anita.
Sci. 31:404.
Wise, M. B., T. N. Blumer, and E. R. Barrick. 1964.
Effect of various levels of choline on performance
and carcass characteristics of finishing steers fed an
all-concentrate ration. North Carolina Agric. Exp.
Stn. ANS Rep. 139. Anita. Health Ser. 10:22.

Journal of Dairy Science Vol. 67, No. 2, 1984