Agricultural Research Center, Ona * f -"
Research Report RC-1973-4 April 1973 -
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EFFECT OF CAROTENE DEFICIENT RATIONS AND SUPPLEMENTALL : t
VITAMIN A ON PRODUCTION AND LIVER VITAMIN A OF CALVESj j -I
'. .L *- .. ". ** "* c' " '. .:':;..
W1. G. Kirk, R. L. Shirley, J. F. Easley, F. M. Peacock
and Frank G. Martin-'
Vitamin A :icsn essential nutrient for beef cattle.- .It is.involved in
the many physiological activities of other vitamin and" hormones, .protects .
epithelial tissue from infection and- is necessary for-edequate growth., Stress
factors, disease, dittary:deficien.ies, high nitiate level,,, and rich concentrate
rations and high temperatures will increase :the anial.,vitamin A requirements.
Beef cattle obtain most of -their..vi.ta. in A. from carotenes. found in green-
leafy plants.. These carotenes are converted .to vitamin A in the.iintftinal
wall of cattle An excess of.carotene or vitamin. A intake above daily needs .: .
will be stored, in the liver.-as vitamin.-.A.and ,used when needed. Thus the .-
vitamin, and carotene content :of previous and current -rations are important- ,
considerations in. understanding vitamin adequacy in beef production. The- "c ,
objective of three trials was to determine the effect of supplemental .vitamin
A for calves that had been depleted for 140 days on,a ration.low in f protein,
total digestibleinutrients (TDN) and carotene.
1/ Animal Scientist Emeritus, Ons ARC; Animal Nutritionist-and Assistant
.Animal Nutritionist, Main Station, Gainesville; Associate Animal
M;usbandman, One ARC; and Associate Statistician, Main Station, .
Reviews of the known facts of vitamin A in the health of livestock have
been made by Morrison (14), Moore (13), in N.R.C. (16) and Maymone (12). Perry
et al. (17) found that adding vitamin A to a steer ration which met N,R.C.
standards for carotene increased gain and level of liver and plasma vitamin A
but decreased plasma carotene. Kirk et al. (10), show that feeding 30,000 I.U.
vitamin A daily or an initial injection of 2.7 and 5.4 million I.U. to calves
and yearling cattle did not improve rate of gain but increased (P4.,01) liver
vitamin A level. The control cattle did not exhibit any gross symptoms of
vitamin A deficiency other than low rate of gain when vitamin A level of dry
liver was 21 micrograms (mcg) for weanling calves and 5 mcg for yearlings kept
on dry feed for five months before placed on test. Perry et aIlt ( ) increased
production by feeding calves 20,000 I.U. daily or initial injection of 1, 4 and
6 million I.UT vitamin A. Liver storage, however, was not increased with
injection of 1 million I.U. vitamin A. Chapmen et al. () reported that'in two
steer trials gains were not increased by feeding 25,000 I.U. vitamin A daily
while in a third trial gains were 22% above the control.
Kohlmeier and Burroughs ( 'found that no dietary vitamin A was required
for good feed lot performance as long as liver and plasma vitamin A levels
remained above 2 mcg per gram and 25 mcg per.100milliliters (ml1), respectively.
Cattle placed in the feed lot with liver vitamin A from 20 to 40 mcg per gram
will have sufficient tissue reserve for 90 to 120 days under normal conditions.
Cattle with less than 5 mcg per gram of liver would require supplemental vitamin
A almost immediately when placed in the feed lot. They state that the minimal
vitamin A level necessary to maintain the above critical liver and plasma
levels is 636 I.U. per pound of air-dry feed. NRC (15) reported in 1963 that
an adequate calf finishing ration should contain 750 I.U. vitamin A per pound
and in 1970 NRC (16) recommended 1000 I.U.
Ullrey (19) states that recent research show that: (1) chemical analysis
does not establish biopotency of carotene as only 15 of the more than 00
naturally occurring carotenes and carotenoids have vitamin A activity; (2)
presence or absence of the s"ubstancet'in the ration which influence rate of
emulsification and absorption of carotene; and (3) variation in vitamin A
utilization between individual animals of the same species.
Seventy-two weaned calves, 24 in each of three trials with six lots per
trial, were fed a vitamin A deficient ration for 140 days followed by a 56-day
vitamin A supplemental period at the Ona Agricultural Research Center. The
ration ingredients of cottonseed hulls, cottonseed meal, citrus pulp and mineral
mixture, (5 were :elected because they were widely used in mixed Florida
cattle rations. 'Dried citrus pulp furnished from 57.47. to 64.4% by weight
of the total ration.
The following daily additions per animal were made to the basal ration
during the supplemental period: Lot 1, none; Lot 2, 0.5 pounds pangolagrass
hay (hay); Lot 3, 2 pounds hay; Lot 4, 0.5 pounds hay + 12,000 I.U. vitamin
A; Lot 5, 2 pounds hay + 48,000 I.U. vitamin A; and Lot 6, 12,000 I.U. vitamin A.
The rations fed to the six lots of calves in the depletion and supplemental
periods are given in Table 1. The 1963 N.R.C. (5) recommendation for a 600-
pound calf finishing ration should contain 117 protein, 667. TDN and 750 I.U.
vitamin A per pound of feed. In the depletion period the ration fed the six
lots was 5% low in protein, 67. in TDN and 247. in vitamin A. Rations fed in the
supplemental period were from 3% to 7% deficient in both protein and TDN while
the ration fed Lot 1 was 187. and that fed Lot 2 was 127. below the vitamin A
standard and Lots 3, 4, 5 and 6 were 5%, 727., 3497. and 80%, respectively, above
ingredients fed in:depletion and supplemental periods.
I. U. vitamin A
. Supplemental- 56 days
3 4 5 6
Vitamin A requirements % '96 82 88 105' 172 449. 181
Crude protein % 10.5 10.7 10.5 10.2 10.5 10.2 1:0.7
TDN % 62.1 64.2 63.4 61.6 62.6 :61.8 64.3
1/ Vitamin A palmitate, 30,000 I.U. per gram.
_ __ ____
Table 1. Percentage
the standard. 'The calves -ere ed. once.daily as much of the mixed ration as
would be eaten in 24 hours.
A fresh:liver 'sample by the biopsy method () was obtained from each calf
initially and at six 28-day intervals thereafter. A final liver and blood
sample from each animal were taken when slaughtered. A total of 560 fresh
liver samples were assayed for vitamin A and carotene,
The procedure of Gallup and Hoefer (7), modification of the Carr-Price
8bC13 acetic anhydride method, was used to determine the vitamin A and
carotene of liver; that of Kimble (8) for plasma vitamin A and carotene; and
the method outlined by AO.A.C. () for carotene in feed. Individual daily
gain, slaughter data, level of vitamr.n A and carotene in fresh liver collected
at 28-day intervals and final vitamin A and carotene of blood plasma were ;
examined statistically (6.
RESULTS AND DISCUSSION
Production data for the six lots of 12 calves in both the depletion and
supplemental periods each are summarized in Table 2. Average daily gain of the
72 calves fed the same ration in the 140-day depletion period was 1.55 pounds,
a daily gain range of 1.46 pounds for Lot 6 to 1.4$ pounds for Lot 5..
The ration was only moderately palatable as shown by the 2.6 to 2.7 pounds of
feed eaten daily per 100 pounds live weight. Gains were considered relatively
low for calves having a feeder grade of Low Choice. The low gains is attributed
to several factors: deficiency of protein and TDN in the ration; unknown
essential nutrients found in high quality alfalfa meal; and not to a critical
deficiency of vitamin A.
The greater gain by all lots in the supplemental period is to be
expected as calves were larger and more accustomed to their ration and feeding
Table 2. Production data in depletion and
Depletion period, 140 days:
Initial weight, Ib
Daily gain, Ib
TDN/lb gain, lb
Supplemental period, 56 days:
Initial weight, lb
Daily gain, lb
TDN/lb gain, Ib
Ribeye area/100 Lb
carcass, sq. in.
5.6 5.5 6.2
51.1 51.5 51.3 50.6
1 _I_ __ _~ __
routine. Lot 1, control, ate the most feed and had a slightly higher average
daily gain in the 56-day supplemental period than Lots 2 to 6. The addition
of hay, fed lots 2, 3, 4 and 5 reduced level of protein and TDN in their
rations, a probable cause of the lower daily gains. The addition of vitamin
A alone and combined with hay to the basal ration, both at two levels, resulted
in no significant increase in rate of gain and decrease in feed requirements
per unit gain.
No abnormalities were observed in any of the animals at slaughter and
all carcasses were passed by the Federal Meat Inspector. Statistical analysis
did not reveal any significant difference in rate of gain in either the 140-day
depletion period or 56-day supplemental period, for the six lots.
Slaughter and Carcass Results
The average initial feeder grade was "Low Choice", final slaughter grade,
"High Good" and carcass grade "Good" for the 72 calves. There was no signi-
ficant difference in live and carcass grades, dressing percentage, rib eye
area and yield of edible meat for the six lots. Feeder grade indicated a
higher potential than U. S. Good carcass grade if a ration higher in protein
and TDN had been fed. There was no individual or lot data to show that adding
hay and vitamin A to the basal ration improved quality of carcass and increased
the yield of edible meat.
Vitamin A and Carotene in Fresh Liver
Vitamin A. The 72 calves were fed an average of 8093 I.U. vitamin A
daily in the 140-day depletion period, 77%. of the recommended level for
finishing calves having an average weight of 600 pounds. The average vitamin
A level per gram of fresh liver by 28-day periods for the six lots of calves
are given in Table 3. There was considerable initial variation in liver vitamin
b. e , r .,
Table 3. Level of vitamin A in fresh; liver at 28-day intervals.-
,,- ,. i' i *,
Lot 1 2 ..: 3 4
Depletion period 140 days:'
S. fesh live'' : 4/ -
mcg/gram fresh liver- **'
5 : ":6
14 13 ..-
period 56 days:-
Kirk et al. (11)
Fed onty basal ration, 196 days.
Basal 'rat-ion .+ supplements daily per
Lotr 3, 2 b. hay; Lot 4, 0.5 lb hay'+
+4,o000 I.U.; and Lot 6, 12,000 I.U.
., .. . J -,'.
,.. (P/ .01) decrease in
* (P/ .01) increase in
calf; Lot 1, none; Lot 2, 0.5
12,000 I.U. vitamin A; Lot 5,
liver vitamin A, all lots.
liver vitamin A, Lot 5. "
i- ,, ..'. .
lb hay; :
2 lb hay
A between the six lots of calves but these differences became less with each
successive 23-day period. The vitamin A level after C4 days on the depletion
ration for all calves was 78% of the initial value with a further 137. reduc-
tion in the last 56 days, a highly significant (P.O01) rate of decrease.
The highest initial level of vitamin A was 174 mcg per gram of fresh liver
and at the end of the depletion period 32 mcg, both from the same calf. The
lowest individual level at the end of the depletion period was 1 mcg in trial
1, 2 mcg in trial 2 and zero meg in trial 3. These calves, however, appeared
healthy as shown by their appearance and comparable weight gains. It has been
suggested that 23 mcg vitamin A per gram of liver (2) is close to the level
where vitamin A deficiency occurs while the level may be as low as 2 mcg per
gram of liver (9) before animal performance is affected.
There was a further decrease in liver vitamin A in the 56-day supplemental
period for Lot 1, control, and Lots 2, 3 and 4. Feeding 12,000 I.U. daily, Lot
6, maintained liver vitamin A as at the end of the 140-day depletion period.
The data show that liver vitamin A was maintained for this lot when the daily
ration contained 1,353 I.U. per pound; 80% above the NRC (15) recommended level
of 750 I.U. per pound feed. The efficacy of 12,000 I.U. vitamin A daily indi-
cated that part of the 12,000 I.U. fed Lot 4 was required in the digestion of
0.5 pounds low-level carotene hay. Raising the vitamin A level to 3367 I.U.
per pound of feed, the basal ration fed Lot 5 plus 2 pounds of hay and 48,000
I.U. vitamin A daily per calf, resulted in liver vitamin A storage at the same
rate as reduction in last 56 days of the depletion period. A longer than the
56-day period was required to show the effect of the vitamin A supplements on
vitamin A liver storage.
Calves in Lot 1, control, fed a ration 247. below the 1963 NRC vitamin A
recommended level in the depletion and 17% in the supplemental period, showed
no gross symptoms of vitamin A.deficiency. The high average initial level of
113 mcg vitamin A per gram of liver, low rate of gain, 1.58 pounds daily for
the 196 days,.and level of vitamin A in citrus pulp, which made up about 60% of
the total ration eaten, contributed to their well-being in the three 196- day
Carotene. The liver. carotene level for each lot by 28-day intervals is
shown in Table 4. .The highest individual initial level was 43 mcg/gram of
fresh liver with:thwo calves having zero carotene at the end of the depletion
period. Liver carotene was significantly reduced (PL..01); an average of
53% the.first 56 days of the depletion period and 147 in the next 84 days.
The average liver carotene reduction was a further 6% for the 72 calves in
the supplemental period. The supplemental hay and vitamin A at the rates fed
ha4,little if any affect on the carotene level of the liver.
Regressiop..analysis showed that the rate of decline in vitamin A and
carotene in fresh liver during the depletion period differed in the three trials.
Vitamin A and Carotene in Blood Plasma
One blood sample from each calf was obtained at time of slaughter. The
average vitamin A and carotene in blood plasma for-the six lots in each trial
are given in Table 5. The highest individual level of vitamin A was 66 mcg
per 100 ml plasma for a'calf in Lot 5, and the lowest was zero for a calf in
both Lots 1 and 2. The highest individual plasma carotene level was 307 mcg
for a calf in Lot 1 and the lowest was zero for/the same two calves which had
zero plasma vitamin A. Analysis of variance showed that-there was a significant
difference (P-c.0l) in plasma vitamin A between trials and between lots and in
carotene between trials. There was an interaction (P O.-05) between trials and
lots in plasma carotene. Duncan's Multiple range test showed that for the.,
three trials vitamin A in plasma from Lots 4, 5 and 6 was greater (P4.01)
Table 4. Level of carotene in fresh liver at 28-day intervals./
1/ Kirk et al. (11)
2/ Fed only basal ration 196 days.
3/ Supplements fed daily per animal given in Table 3.
** (P/.01) decrease in liver carotene, all lots.
Table 5. Vitamin A and carotene in
per 100 ml plasma
Avg 3 trials
Avg- 3 als 123
Avg 3 trials 123
abc means in the same row or
column with different superscripts differ
- -- --
- -- ---
than for Lots 1, 2 and 3 but no significant difference was detected within
each of the two groups.
There were no significant differences in plasma carotene among the lots
in Trial 1. Lots 3 and 5 had significantly less p.lama: carotene than Lots 1
and 2 in Trial 2. Plasma carotene was higher (P-.0Ol) for Lot 3 trial 3 than
for the other five lots, with no difference among Lots 1, 2, 4, 5 and 6.
Supplemental feeding of hay at two levels-.in combination with Vitamin A
at two levels and vitamin A alone increased plasma vitamin A for Lot 4, 5 and
6; only Lot 5, however, had an increase in liver vitamin A (Table 3) and the-
lowest plasma carotene. Apparently Lot.5 fed 2 pounds of hay and 48,000 I.U.
vitamin A daily converted more of the ration carotene to vitamin A with
excess stored in the liver as vitamin A.
Avergge.idaily. gain for the 72 calves was 1.55 pounds in the 140-day
depletion period and 1.05 pounds in the 56-day supplemental period, low for
calves having an initial feeder grade of Low Choice. All rations were low in
protein and TDHNand unknown nutritional factors found in alfalfa leaf meal to
promote rapid.gain. There was ;no significant difference in rate of gain be-
tween the six lots of calves in either the depletion or supplemental periods.
Calves fed the control ration for 196 days showed no apparent gross vitamin A
deficiency symptoms. "
Vitamin A per gram of fresh liver was reduced from an average of 105 mcg
initially to Smcg:Gfter 140 days on the basal ration, a 917. decrease (P/_.01),
There was a further small decrease in liver vitamin A in the 56-day supplemental
period ; for Lpts 1, .2, .3 and 4 with 'Lot 6 at the same level at the end of
48,000 I.U. vitamin A daily per animal (3367 I.U. vitamin A per pound of feed)
increased from 6 to 17 mcg per gram, the same rate of increase as decrease
in the last 56 days of the depletion period.
Carotene per gram fresh liver was reduced from an average of 15 mcg
initially to 5 mcg in the 140-day depletion period, a 677 decrease (PL..01)
with little change in liver carotene values in the 56-day supplemental
period for any lot.
.The addition of hay and vitamin A, both at two levels, (Lots 4 and 5)
and vitamin.A alone (Lot 6) in the 56-day supplemental period increased
vitamin A. level of.:blood plasma .P i01)'. Lot 5 calves fed the basal.ration
plus 2 pounds hay and 48,000 I.U. vitamin A daily had the lowest level of
plasma carotene and the highest level of liver vitamin A.
1. A.O.A.C. 1960. Official methods of analysis (9th ed.). Association of
Official Agricultural Chemists, Washington, D. C.
2. Beeson, W. M., T. W. Perry, W. H. Smith, and M. T. Mohler. 1965. Effect
of different levels of vitamin A on the performance of Hereford and
Charolais x Hereford heifers self-fed on blue grass pasture. Purdue
Univ. Agr. Exp. Sta. Res. Proj. Rept. 169.
3. Chapman, H. L. Jr., D. H. Cox, C. E. Haines, and G. K, Davis. 1963.
Evaluation of the liver biopsy technique for mineral nutrition studies
with beef cattle. J. Ani. Sci. 22:733-737.
4, Chapman, J. L. Jr., R. L. Shirley,'A. Z. Palmer and J. W. Carpenter. 1971.
Vitamins A and E. Fla. Agr. Exp. Sta. I.F.A.S, Bull. 748.
5. Cunha, T. J., R. L. Shirley, H. L. Chapman, Jr., C. B. Ammerman, G. K. Davis,
W. G. Kirk, and J. F. Hentges, Jr. 1964. Minerals for beef cattle in
Florida. Fla. Agr. Exp. Sta. Bull. 683.
6. Duncan, D. B. 1955. Multiple range and multiple F-test. Biometrics 11:1.
7. Gallup, W. G. and J. A. Hoefer. 1946. Determination of vitamin A in
liver. J. Ind. Eng. Chem. 18:288.
8. Kimble, M. S. 1939. The photoelectric determination of viteain A and
carotene in human blood plasma. J. Lab. Clin. Med. 24:1055.
9. Kohlmeler, R. H. and Wise Burroughs. 1970. Estimation of Critical
plasma and liver vitamin A levels in feedlot cattle with observations upon
influence of body stores and daily dietary requirements. J. Attim; Sci.
10. Kirk, V. G., R. L. Shirley, J. T. Easley, and F. M. Peacock. lS70, Cattle
gains fed and injected with vitamin A. Fla. Agr. Exp. Sta. Tech Bull. 741.
11. Kirk, H. G., R. L. Shirley, J. T. Easley and F. M. Peacock. 1971. Effect
of carotene deficient rations and supplemental vitamin A on gain, feed
utilization and liver vitamin A of calves. J. Anim. Sci. 33:476-4;0.
12. Maymone, B. 1965. Connaissance sur 1' activeit E biochemique de 1A
vitamin A et son importance pour l'utilivsation des vitamins conte8us
den les ailments compose's pour animaux. World Review of Animal Production.
Rome, Italy, July-Sept. 99-123.
13. Moore, Thomas. 1957. Vitamin A. Elsevier Publishing Co., New York.
14. Morrison, F. B. 1956. Feeds and Feeding. 22nd Ed. The Morrison Publishing
Company, Ithaca, N. Y.
15. Nutrient requirements of beef cattle. 1963. National Res. Council Publ.
1137. Washington, D. C.
16. Nutrient requirements of beef cattle. 4th Rev. Ed. 1970. National
Res. Council. Washington, D. C.
17. Perry, T. W., W. M. Beeson, M. T. Mohler and W. H. Smith. 1962. Level
of supplemental vitamin A with and without suncured alfalfa meal for
fattening steer calves. J. Anim. Sci. 22:333-339.
MAR 6 utw
18. Perry, T, W.,, M. Beeson,. H. H. Smith and M. T. Mohler. 1967.
Injectable vs oral vitamin A for fattening steer calves. J. Anim.
19. Ullrey, D. E. 1972. Biological availability of fat-soluble vitamins:
Vitamin A and carotene. Jour. An. Sci. 35:648-657.
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