Siciliano 1993 Aceite Soya Vitamina E Caballos

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P. D. Siciliano and C. H. Wood

The effect of added dietary soybean oil on vitamin E status of the horse

1993. 71:3399-3402. J Anim Sci

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The Effect of Added Dietary Soybean Oil on

Vitamin E Status of the Horse

P. D. Siciliano and C. H. Wood'

Department of Animal Science, University of Kentucky, Lexington 40546

ABSTRACT: Fourteen 2-yr-old Quarter Horses and

Quarter Horse x Thoroughbreds were randomly as-signed to either a control (CTRL) diet o r a diet

supplemented with 6.4% soybean oil ( S B O ) . The

amounts of both diets that were fed met current NRC

nutrient requirements and were isoenergetic with

similar nutrient:energy ratios. Venous blood samples

(20 mL) were taken at d 0, 30, 60, and 90 of the

experiment and analyzed for serum a-tocopherol,

serum cholesterol, and serum triglyceride. The sum of

serum cholesterol and serum triglyceride values was

used as an estimate of serum total lipid. The ratio ofserum a-tocophero1:serum total lipid was used as a

measure of vitamin E sta tus of horses. The SB O group

had higher serum cholesterol concentrations ( P <

.001) a t 30, 60, and 90 d. Serum triglyceride was not

affected by diet. Serum total lipids tended t o be

greater ( P < .06) for the SBO group. Serum a-

tocopherol was greater ( P <.001) at 90 d in the SBO

group. There was a significant correlation ( P<.001, r

= .62) between serum total lipids and serum a-

tocopherol concentrations in the SB O group. This

correlation was not significant ( P <.07, r =.35) in the

CTRL group. The ratio of serum a-tocophero1:serum

total lipid was not affected by diet. The results of this

experiment suggest that vitamin E status in non-

exercising, 2-yr-old horses fed currently recommended

levels of vitamin E was not affected by the addition of

6.4% SBO to the diet.

Key Words: Horses, Vitamin E, Polyenoic Fatty Acids, Cholesterol, Lipids

Introduction

The use of added dietary fat as a source of energy

for horses has become increasingly popular in recentyears. Two commonly used sources of dietary fat are

corn oil and soybean oil ( SBO). Both corn oil and

SBO contain large amounts of polyunsaturated fatty

acids (PUFA). Diets containing added fat high in

PUFA (e.g., corn oil, soybean oil, and cod liver oil)

have been shown t o decrease vitamin E status, which

resulted in vitamin E deficiency symptoms in chicks

(Dam, 19621, ducks (Jager, 19721, dogs (Hayes et

al., 19681, swine (Grant, 19611, calves, and lambs

(Blaxter, 1962). There have been several studies in

horses concerning the utilization of dietary fat as a

source of energy (reviewed by Lawrence, 1990);

however, none has investigated the effect of fats,specifically those high in PUFA, on vitamin E status

of the horse. Vitamin E has been shown t o affect

immunocompetence of horses (Baalsrud and Overnes,

19861, in addition t o providing protection against

'To whom correspondence should be addressed

Received December 7 , 1992.

Accepted August 16, 1993.

J. Anim. Sci. 1993. 71:3399-3402

muscular dystrophy in foals (Wilson et al., 1976;

Schougaard et al., 1972). The following study was

designed t o investigate the effect of added dietary SBO

(6.4%of the total d ie t) on vitamin E status of horses.

Materials and Methods

Fourteen 2-yr-old horses, of Quarter Horse and

Quarter Horse x Thoroughbred breeding, were ran-

domly assigned t o one of two dietary treatments:

control (CTRL) n = 7 ) or SBO-supplemented (n =

7 ) . Treatment groups were balanced for age and sex.

Mean BW were similar in both treatment groups at

th e start (520 f 10 kg) , and finish (CTRL, 530 f 7 kg;

SBO, 529 -t 12 kg) of the study. Both treatment

groups of horses were fed a ration composed of a grain-based concentrate and alfalfa hay, which was formu-

lated to meet NRC (1989) requirements (Tables 1

and 2) . All ingredients in both diets were analyzed fo r

ADF, CP, calcium, phosphorus, and selenium (Chemi-

cal Service Laboratory, Louisville, KY). Digestible

energy of ingredients was calculated according t o the

equat ion of Fonnesbeck (198 1). Vitamin A concentra-

tion of ingredients was calculated using NRC (1989)

table values. All ingredients in both diets were

3399







3400 SICILIAN0 AND WOOD

Table 1. Composition of control (CTRL) and addeddietary soybean oi l diets (SBO)ab

Ingredient CTRL SB O

Oats, kg/d 2.90 .96

Soybean meal, kg/d - .46

Soybean oil, kgld - .50

Trace minera l sal t, g/dC 14.50 14.50

Vitamin E supplement, g/dd 3.74 3.62Alfalfa hay, kg/d 5.90 5.90

a l O O% dry matter basis.

bDuring the last 30 d of the trial, a new lot of hay with slightly

different nu tri ent composition (compared with hay fed durin g d 0 to

6 0 ) was fed. To maintain previous nutr ien t levels, the amount ofhay was decreased from 5.9 to 5.6 kg/d. The vitamin E supplement

was also adjusted to maintain vitamin E levels fed d 0 to 60.

CComposition of trace mineral salt : NaCl 5 98.59’~~ 96%; 35 mg/

kg of Zn as ZnO; 34 mg/kg of Fe a s Fe2O3; 20 mg kg of Mn as MnOZ;

3.3 mgkg of Cu as CuO; .7 mg kg of I as Ca(I03)2; .5 mg kg of Co as

d176,000 IU of vitamin E activitykg supplied as all-rue-a-

tocopheryl acetate. Manufactured by Carl S. Akey, Lewisburg, OH.

coco3 .

analyzed for vitamin E activity (Animal Health

Diagnostic Laboratory, Lansing, MI). The SBO diet

differed from the control diet in tha t it contained 6.4%

SBO in the total ration (25% in the concentrate

portion of the rat ion) . In addition, soybean meal was

added to the SBO diet so that both treatments were

isonitrogenous. Both SBO and CTRL rations were

isoenergetic and similar nutrient:energy ratios were

maintained. Vitamin E supplement containing

176,000 I U k g provided by all-rac-a-tocopheryl aceta te

was top-dressed on the concentrate of each individual

horse’s ration at each feeding, so that total vitamin E

intake, which included vitamin E activity from theconcentrate, hay, and vitamin E supplement, was

equal in both treatment groups. All horses were

individually fed one-half their respective rations twice

daily. Both rations were readily consumed by all

horses. Horses were individually housed in open-air

pens that were partially covered for shelter.

Blood samples (20 mL) were taken via venipunc-

ture at 0, 30, 60, and 90 d of the experiment, before

the afternoon feeding, and refrigerated ( 3 t o 5°C)

immediately after collection. Serum was separated

within 4 h of collection, and stored a t -4°C. Serum

samples were analyzed fo r a-tocopherol, cholesterol,

and triglycerides. Alpha-tocopherol was measured

using HPLC with a fluorescence detector according to

the method of Hidiroglou (1989). Cholesterol and

triglycerides were measured by colorimetric tech-

niques using commercial kits (Sigma Diagnostics, St .

Louis, M O ) . A lipid control (Sigm a Diagnostics) was

used with both the cholesterol and triglyceride kits t o

verify the procedure and reagent reliability.

Cholesterol and triglyceride concentrations (milli-

gramddeciliter ) in the serum were added to estimate

serum total lipids according t o the method of Thurn-

ham et a l. (1986) . Serum a-tocopherol was expressed

in a ratio with serum total lipid (milligrams of a-

tocophero1:grams of serum total lipid) t o represent

vitamin E status more accurately, according t o th e

method of Horwitt et al. (1972).

Variation in response variables (serum a-

tocophero1:serum tota l lipid [milligrams/graml, serum

a-tocopherol [micrograms/milliliterl,serum total lipids

[milligrams/deciliter] , serum cholesterol [milligrams/

deciliter], and serum triglycerides [milligrams/decil iter]) were partitioned using ANOVA for repeated

measures designs (SAS, 1985). Differences between

treatments were analyzed using between-horse varia-

tion as the error term. Differences due t o period and

period x treatment interaction were analyzed using

within-horses variation. Treatment difference were

considered significant at the P < .05 level. Where

period effects were significant ( P < .05), orthogonal

polynomials were used t o determine the response

trend. Where period x treatment interactions were

significant (P <.05),a LSD test was used as a means

separation technique.

Results and Discussion

Serum a-tocopherol concentrations in the present

study in both treatment groups increased (linear; P <

.001) above the concentrations measured at the

beginning of the study (Table 3 ) . The increase may

have been due t o feeding higher amounts of vitamin E

during the study as compared t o the amount fed before

the experiment. There was a significant diet x period

interaction (P < .03) on serum a-tocopherol. The

concentration of a-tocopherol in the SBO group tendedt o be greater relative t o the CTRL group at 60 d, and

at 90 d the a-tocopherol concentration in the SBO

group was greater ( P <.001) than that in the CTRL

group (Tab le 3). Serum to tal lipids also tended t o be

greater ( P < .06) in the SBO group (Table 3 ) . The

increase in a-tocopherol concentrations in the horses

fed the SBO diet was correlated ( P < .001; r = .62)

with an increase in serum total lipids. Correlations

between tocopherols and total lipids have been

reported in both rat and human studies (Horwitt et

Table 2. Nutrient intake of control (CTRL) andadded dietary soybean oil (SBO) dietsa

Nutrient CTRL SB O

DE , Mcal/d

CP, kg/d

Ca, gid

P, gld

Se, mgid

Vitamin A, IUld

Vitamin E, IU/d

22.08 22.01

1.37 1.37

57.00 56.00

26.00 22.00

2.60 2.00

153,447

720 720

153,542

aAs-fed basis.







VITAMIN E STATUS OF THE HORSE

Table 3. Serum concentrations of cholesterol, total lipids, 0-tocopherol,and a-tocophero1:total lipids in horses fed control diets (CTRL) and

soybean oil-supplemented diets (SBO)”

3401

Days~

Item 0 30 60 90 SE

Cholesterol, mg/dLbC

CTRL

SB O

CTRL

SBO

CTRL

SBO

CTRL

SB O

Total lipids, rng/dLd

n-Tocopherol, pg/rnLde

wTocophero1:total lipids ,

84.99

89.60

118.00

132.30

3.80

4.00

3.30

3.00

mdg‘

85.9Eif

107.478

129.20

157.90

5.00

4.90

3.90

3.10

89.05f

114.119

135.50

167.30

4.90

5.50

3.70

3.30

81.21f

111.298

123.70

161.90

4.70f

6.109

3.90

3.80

3.81

3.81

5.12

5.12

.25

.25

.20

.20~ ~ ~ ~

aBoth diets were fed in amounts to meet NRC requirements for 2-yr-old, nonexercising horses. Both

rations were isoenergetic with similar nutr ient to energy ratios. The SBO diet contained 6.4% soybean oil.

bMain effect of diet ( P < ,041; diet x period interaction ( P < .01).

CLinear period effect ( P < .01).

dLinear period effect ( P < .001).

eDiet x period interaction I P < ,031.f,gMeans within an item within a column lacking a common superscript letter differ ( P < .001).

al., 1972). The correlation between serum CY-

tocopherol and serum total lipids was much stronger

in the SB O group ( P<.001; r =.62) than in the CTRL

group ( P<.07; r =.35).This is in agreement with the

report of Bieri (1990), who found the correlation

between serum tocopherols and serum total lipids to

be much weaker in normolipemic subjects. It is

thought that as serum total lipids increase, vitamin Emoves out of tissues and into circulating high-density

lipoproteins (Bieri et al., 1977). This migration ofvitamin E from tissues t o circulating high-density

lipoproteins in association with increased serum lipids

may explain the increased serum a-tocopherol in the

SBO group in the present study. It is also possible

that the added fat in the SBO diet may have increased

vitamin E absorption. Vitamin E absorption is closely

associated with fat absorption (Bie ri, 1990 ), and

increasing the amount of added dietary fat in the diet

of ponies has been shown to increase the digestibility

of ether extract (Ka ne e t al., 1979); therefore, added

dietary fat may enhance vitamin E absorption.

There was a significant effect of diet ( P<.04) and

a significant diet x period interaction ( P < .01) on

cholesterol. At 30, 60, and 90 d cholesterol concentra-

tions were significantly greater ( P <.001) in the SB Ogroup than in the CTRL group (Table 3 ) . The higher

concentration of serum cholesterol in the horses fed

the SBO diet resulted in the tendency toward in-

creased concentrations of serum total lipid in the

horses fed the SBO diet, as discussed previously.

Increases in serum cholesterol in horses and ponies

fed added dietary fat have been reported in other

studies (Bowman et a l., 1977; Hambleton et al., 1980;

Kurcz et al., 1991 ). It i s possible that the increase in

serum cholesterol in the horses fed the SBO diet was a

result of an increase in cholesterol-containing

lipoproteins needed for fat transport.

Serum cholesterol (Table 3 and serum triglyceride

(d at a not shown) concentrations increased (l inea r, P

< .01) throughout the experimental period in both

treatment groups. This increase in serum cholesterol

and triglyceride concentrations contributed to a sig-

nificant linear effect of period (P .:.001) in serum

total lipids (Table 3 ) . Increasing concentrations ofserum total lipids in both groups may reflect synthesis

of excess energy metabolites into triglycerides for

storage. Both groups of horses were assumed t o be in a

positive energy balance because they gained approxi-

mately .1 kg/d throughout the experiment.

Vitamin E status, measured by the ratio of serum

a-tocophero1:serum total lipids increased (linear, P <

.O 1)above the ratios measured a t the beginning of the

study (Table 3 ) . The increase may have been due t o

differences in vitamin E content of diets fed before the

experiment as compared with experimental diets.

Serum a-tocophero1:serum total lipids w as not

affected by the addition of SBO t o the diet in the

present study (Table 3) . Feeding diets high in PUFA

can increase the amount of PUFA in cell membrane

phospholipids (Rice and Kennedy, 1988) . Membranes

containing large amounts of PUFA in their phos-

pholipids are especially susceptible t o lipid peroxida-

tion (Combs et al., 1975 ). Vitamin E is a biological

antioxidant that protects cell membranes against

damage from lipid peroxidation (Combs et al ., 1975;

Chow, 1991). Increasing PUFA content of tissues has

been shown t o increase vitamin E utilization in rats

fed corn oil as a source of PUFA (Weiser and Salkeld,







3402 SICILIAN0 AND WOOD

1977). The absence of any effect of SBO supplementa-

tion on vitamin E statu s in this experiment suggests

tha t the level of vitamin E in the diet was adequate t ocompensate for the additional PUFA content of the

diet. Hayes et al. ( 19 68 ) reported no sign of decreased

vitamin E sta tus in vitamin E-supplemented dogs fed

graded levels of PUFA; however, the vitamin E status

of dogs fed diets containing PUFA that were not

supplemented with vitamin E decreased with increas-

ing PUFA. In addition, the level of PUFA in this study

may not have been high enough t o interfere with

vitamin E. Jager (19721 found interference between

PUFA and vitamin E in rats only at high dietary

were also highevels of fat (approximately 15%)&at

in PUFA (approximately 50%).

Implications

The results of the present study

soybean oil fed at 6.4% of the total

interfere with vitamin E status in

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D. Lewis. 1980. Dietary fat and exercise conditioning effect on

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Hayes, K. C., S. W. Nielsen, and J. E. Rousseau, J r. 1968. Vitamin E

deficiency and fat stress in the dog. J. Nutr. 99:196.

Hidiroglou, M. 1989. Mammary transfer of vitamin E in dairy cows.

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Horwitt, M. K., C. C. Harvey, C. H. Dam, Jr., and M. T. Searcy.

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indicate that

nonexercising,

diet does not

2-yr-old horses fed currently recommended levels ofvitamin E. These results may not be applicable to the

exercising horse fed soybean oil.

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