Professional Documents
Culture Documents
net/publication/19784714
CITATIONS READS
83 161
3 authors, including:
Some of the authors of this publication are also working on these related projects:
All content following this page was uploaded by Gerald B Huntington on 14 May 2014.
The online version of this article, along with updated information and
services, is located on the World Wide Web at:
http://jas.fass.org/content/66/2/342
www.asas.org
J. H. E i s e m a n n , G. B. H u n t i n g t o n a n d C. L. Ferrell 2'3
U S D A - A R S , R o m a n L. H r u s k a U.S. M e a t A n i m a l R e s e a r c h C e n t e r
Clay Center, NE 6 8 9 3 3
and
Beltsville A g r i c u l t u r a l R e s e a r c h C e n t e r
Beltsville, MD 2 0 7 0 5
ABSTRACT
The objective of this study was to measure acute (d 1) and chronic (d 9) effects of dietary
clenbuterol on heart rate, blood flow, oxygen uptake, and net uptake/release of metabolites in the
hindquarters of growing steers. The design was a single reversal with two 9-d periods of control or 8
mg clenbuterol/d with 5 d between periods. Within 2 h of initial consumption of 2 mg clenbuterol
(d 1), heart rate and blood flow doubled and arterial plasma concentrations of glucose, L-lactate
and nonesterified fatty acid (NEFA) increased, whereas cx-NH2 N and NH 3 concentrations decreased,
demonstrating an acute response. Uptake of oxygen increased and net uptake of cz-NH2 N de-
creased. Net release of both L-lactate and NEFA increased. On d 9, there were no acute responses
to clenbuterol consumption; however, heart rate, blood flow, and NEFA concentration remained
chronically elevated, and plasma concentrations of acetate and propionate decreased compared
with control feeding. Net uptake of c*-NH2 N, oxygen and release of L-lactate by the hindquarters
chronically increased on d 9 of clenbuterol feeding. Changes in both blood flow and arteriovenous
(AV) concentration difference contributed to changes in uptake/release. The chronic metabolic
changes and oxygen uptake were consistent with increased N retention in the hindquarters through
increased protein synthesis, decreased use of acetate and increased reliance on NEFA for cellular
energy. In conclusion, the data show that the perturbation of homeostatic regulation by dietary
clenbuterol on d 1 evolved to establishment of homeorhetic regulation by d 9 that is consistent
with increased skeletal protein accretion in growing steers.
(Key Words: Cattle, Blood Flow, Metabolism, Regulation.)
Introduction m a l in such a w a y t h a t a c c r e t i o n of m u s c l e or
C h a n g e s in m e t a b o l i c r e g u l a t i o n a n d n u t r i e n t a d i p o s e tissue is s u b s t a n t i a l l y c h a n g e d . T h e r e
p a r t i t i o n i n g d u r i n g g r o w t h are s u b t l e a n d t h e r e - are a n u m b e r of a g e n t s c a p a b l e o f e f f e c t i n g
f o r e are d i f f i c u l t t o m e a s u r e or assess. O n e w a y such changes, t h e m o s t d r a m a t i c o f w h i c h are
to study the regulation of nutrient partitioning c o m p o u n d s k n o w n collectively as /3-adrenergic
is t o p e r t u r b t h e p h y s i o l o g y o f t h e g r o w i n g ani- agonists. T w o o f t h e s e c o m p o u n d s , c l e n b u t e r o l
a n d c i m a t e r o l , are orally active a n d are effective
in m a n y species i n c l u d i n g pigs, s h e e p a n d c a t t l e
( B a k e r et al., 1 9 8 4 ; D a l r y m p l e et al., 1 9 8 4 ;
R i c k s et al., 1 9 8 4 ; H a n r a h a n et al., 1986). A t
levels t h a t did n o t i n h i b i t gain, c l e n b u t e r o l feed-
1Mention of a trade name, proprietary product, or
specific equipment does not constitute a guarantee or ing increased p r o t e i n c o n t e n t of t h e 9 t h to 1 1 t h
warranty by the USDA and does not imply approval rib s e c t i o n b y 13% a n d d e c r e a s e d fat c o n t e n t
to the exclusion of other products that may be suitable. b y 20% in cattle (Ricks et al., 1 9 8 4 ) . Even m o r e
2The authors thank B. Larsen, L. Laaker, and K.
d r a m a t i c carcass c h a n g e s were o b s e r v e d in cattle
Sorensen for feeding and care of the steers; M. Buschow
and R. Jaeger for assistance at surgery; C. Felber, D. in r e s p o n s e to c i m a t e r o l ( H a n r a h a n et al., 1 9 8 6 ) .
Robertson, J. Whitt, and E. Zetina for technical sup- O f i n t e r e s t are t h e u n d e r l y i n g m e t a b o l i c
port; and J. Rosch for typing the manuscript. changes a n d c o n t r o l s t h a t m u s t b e a l t e r e d to
3The authors with to thank Boehringer lngelheim b r i n g a b o u t t h e carcass changes previously cited.
Animal Health, Inc., for providing the clenbuterol used
in this study. T o d a t e , t h e m e c h a n i s m s r e s p o n s i b l e for a l t e r i n g
Received April 1, 1987. n u t r i e n t p a r t i t i o n i n g in r e s p o n s e to/3-adrenergic
Accepted August 13, 1987. agonists are n o t well d e f i n e d . T h e o b j e c t i v e s o f
on ice. Aliquots of whole blood were removed use of heparin to flush catheters was not alter-
for later analyses, packed cell volume (PCV) ing NEFA concentrations.
was determined, and the samples were centri- A transit-time ultrasonic blood-flow meter a
fuged at 1500 • g for 20 min at 4 C to obtain (Model T101) was used to measure blood flow
plasma. Plasma was aliquoted and stored at - 2 0 and heart rate at the time of each blood sample.
C. Integrated average blood flow rates monitored
Total oxygen was determined on individual at 10-s intervals were averaged for 5 min, en-
heparinized whole blood samples, sealed in 1-ml compassing the blood sampling interval, to ob-
syringes until analyzed with a Lex-O2-Con K tain a single value for blood-flow rate at each
oxygen analyzer s . Equal aliquots of blood were blood sampling interval.
pooled within arterial or venous source and steer Calculations. Plasma flow was calculated
on sampling days for determination of volatile using the following equation:
fatty acids (VFA) by the method of Reynolds
et al. (1986). Similarly, for analysis of D-3-hy- Plasma flow (L/min) =
droxybutyrate, 1 ml of whole blood from each
sample was combined with .1 ml of 6N per- (1 - (PCV/100)).Blood flow (L/rain).
chloric acid and mixed. After completion of
sample collection, the acidified whole blood Net uptake or release of nutrients was calcu-
was centrifuged at 12,000 x g for 30 min at 4 lated as the product of arteriovenous (AV)
C. The supernatant fraction was neutralized concentration difference and whole blood (or
with 6N KOH and centrifuged at 1500 x g for plasma) flow.
10 min at 4 C. The supernatant fraction from For variables analyzed on individual samples,
the second centrifugation was stored frozen mean values were calculated for each steer and
until analyzed by the method of Williamson sample day. For each variable, the mean values
and Mellanby (1974). from each steer during the control and treated
Plasma concentration of glucose was mea- periods were used to calculate a paired t-sta-
sured by a glucose oxidase method 6 modified tistic for d 1, to evaluate acute response to
to use 3,3r-dimethoxybenzide-dihydrochlOride clenbuterol. The variation among animal means
as the coupling agent, c~-amino N (a-NH2 N, was used to test the effect of treatment. The
leucine equivalent) by the method of Broderick same procedure was used with d-9 data to
and Kang (1980) modified to include sample evaluate chronic response to clenbuterol.
dialysis, urea by a diacetylmonoxine method 7
and ammonia by a hypochlorite method s on Resu Its
individual samples b y automated procedures. Steers consumed all feed offered during
Plasma concentration of L-lactate was deter- measurements. On d 1 of clenbuterol feeding,
mined with a membrane-immobilized enzyme steers consumed the 0700 feeding in approxi-
system involving lactate oxidase 9. Plasma con- mately 15 min. Following the initial feeding,
centration of nonesterified fatty acids (NEFA, steers consumed little feed for the remainder of
palmitate equivalent) were analyzed on a pooled the 24-h period. Steers gradually resumed
sample using an enzymatic assay 1~ The kit consumption over the next 4 d. Average daily
instructions were modified by diluting reagents intake was 3.87 + .09 kg dry matter (712 g
1 + 1 with deionized water, adding one-half the crude protein and 11.46 Mcal metabolizable
volume of reagents specified and incubating for energy)/steer. Over the 23-d experimental
20 min. Recoveries were 94.1% (SE = .4%). period, steers gained 20 + 2 kg body weight.
Analysis of individual samples indicated no Initial consumption of clenbuterol caused a
trend toward increased NEFA concentrations rapid doubling of heart rate (figure la). Heart
throughout sampling during control or clenbu- rate decreased slightly from this peak but
terol periods (data not shown), indicating that remained elevated and relatively constant
throughout the duration of sampling on d 1.
There was some fluctuation in heart rate after
control feeding (figure la), but no dramatic
s Lexington Instruments, Waltham, MA. changes. Blood flow showed a response simi-
#Technicon Industrial Method No. 566-79T, Tech-
nicon Industrial Systems, Tarrytown, NY. lar to that of heart rate (figure la). Mean values
7Technicon Industrial Method No. 339-01. of both heart rate and blood flow were increased
e Technicon Industrial Method No. 337--74T. (P<.01) with clenbuterol compared with
D0y I Doy 9
(figure 5a). Arterial c o n c e n t r a t i o n o f urea was
180
B n o t a f f e c t e d (figure 3a). Mean arterial c o n c e n -
i60
3E t r a t i o n s o f glucose ( P < . 0 2 ) , L-lactate ( P < . 0 1 )
~ i40
i
/ and N E F A ( P < . 0 1 ) w e r e higher, c o n c e n t r a t i o n s
<~ 120
I o.S~'-o""o-o-or O-o-O-o.o o f t~-NH2 N ( P < . 0 5 ) and NH 3 (P<.O1) w e r e
I00
lad l o w e r and c o n c e n t r a t i o n s o f O2, acetate,
~ a0 propionate, D-fl-hydroxybutyrate and urea
~ 60 w e r e n o t altered o n d 1 o f c l e n b u t e r o l com-
<
~ 40 p a r e d w i t h c o n t r o l f e e d i n g (table 2). On d 9,
' ' ' I I I t h e r e w e r e no a c u t e changes in arterial c o n c e n -
t r a t i o n o f any o f t h e m e t a b o l i t e s m e a s u r e d
i 16 (figure 2b t o 5b) following c l e n b u t e r o l feeding.
14 F u r t h e r , t h e r e were no c h r o n i c r e s p o n s e s t o
J
12 ,o.-O..o-O'~176 ~ c l e n b u t e r o l in m e a n arterial c o n c e n t r a t i o n s o f
iO
/ o-~a-o-o- o-o.o_o.o.o-o
e ,~
o
0
m 4
1700 0900 1100 1300
'
0700
' '
0900
' I
IlO0
I
13100
6.6 f ~
T I M E OF D A Y ( h ) T I M E OF D A Y ( h ) 6.4 / '~
E 6,2 ~' ~ ~Q
Figure 1. Heart rate and blood flow to the hind-
o 6.0 b.Cl
quarters in steers at each sampling time. Sampling
J
began at 0700 and continued for 6 h at 30-min inter- 5.8
vals. The arrow indicates time of feeding. A: Samples 5.1B
b..o.~..o,.o-O
< 1.0
w
c o n t r o l feeding o n d 1 (table 2). Packed cell
I I I i i I I
v o l u m e was also increased ( P < . 0 5 ) o n d 1 o f
clenbuterol compared with control feeding 20
+ +
r
(table 2). By d 9 o f c l e n b u t e r o l feeding, a c u t e
changes in heart rate and blood flow were no _= 16
1
=0 I
longer observed, following consumption of E
clenbuterol at the beginning of the sampling IZ
0700
, ,
0900
, ,
Im~
, ,
1300
feeding (figure l b and table 2). T I M E OF DAY ( h ) T I M E OF D A Y ( h )
O n d 1, c l e n b u t e r o l f e e d i n g caused a transi-
Figure 2. Arterial 02 concentration, arteriovenous
ent increase in arterial concentration of O2 (fig- concentration difference and 02 uptake in steers at
ure 2a), a progressive increase in arterial concen- each sampling time. Sampling began at 0700 and con-
tration o f glucose (figure 3a) and L-lactate tinued for 6 h at 30-min intervals. The arrow indicates
(figure 4a) and a progressive decrease in arterial time of feeding. A: Samples taken on d 1 of control
( e - - e ) or clenbuterol (o--o) treatment. Average SEM
c o n c e n t r a t i o n o f NH 3 (figure 3a) and a-NH2 N for control and clenbuterol treatments, respectively,
was: Arterial 02 (raM) .20 and .21; arteriovenous 02
(mM) .23 and .23; 02 uptake (mmol/min) 1.07 and
2.01. B: Samples taken on d 9 of control ( e - - e ) or
clenbuterol (o--o) treatment. Average SEM for control
9yellow Springs Instrument Co., Inc., Yellow and clenbuterol treatments, respectively, was: Arterial
Springs, OH. 02 (raM) .16 and .20; arteriovenous 02 (mM) .17 and
l~ Chemicals USA, Dallas, TX. .23; 02 uptake (mmol/min) 1.86 and 2.25.
8
. . z~ ~ z ~
~o
~z .u
b.,
~ ~ ~ . . . .
Z
z~
Om <
-6
~z
ooo ZZZZ . . . . .
<
Z
0
m~
ea~
eL
Om
~Z
[...,
Z .~ o,
.. B.-~
e~
gE'~
r A ,o
18
~E .3
5t H
tl
z 4F ,o,"~
.I
,= :t/
. . . . . I
o'oo'o~o' ,~o',r o,oo o,0o ,,oo ,~o
TIME OF DAY(h) TIME OF DAY(h)
1
Figure 3. Arterial plasma glucose, urea-N and o
NH 3-N concentrations in steers at each sampling time.
Sampling began at 0700 and continued for 6 h at 30-
min intervals. The arrow indicates time of feeding. A:
Samples taken on d 1 of control ( e - - e ) or clenbuterol
(o . . . . o) treatment. Average SEM for control and clen-
buterol treatments, respectively, was: Glucose (mM)
.25 and .89; urea-N (mM) 1.0 and 1.3; NH~ (raM)
.019 and .006. B: Samples taken on d 9 of control
+!
t
( o - - e ) or clenbuterol (o--o) treatment. Average SEM
for control and clenbuterol treatments, respectively,
was: Glucose (mM) .25 and .22; urea N (mAd) 1.0 and
1.5 ; NH 3 (mM) .023 and .014.
!!'
-4
J I 9 . . . . . . J . i i
m~
Z
0
F-
<
I
<
r~ -1
z
d
[..,
< . . . . . ~ o ~
,4
Z
Z
0 8
#.
<
8~
-4
0 I
g "~ X
,4 4 9 9 9 ~ "~ ~ 44
I
II
..~ ~ II
o ~, o
A .B
-~.0
1.8
e
Z 1.6
1,4
III I.Z t \ t
E-,
Z
~ .. .
~t
.1 <
I i I i . . . . I I I
[.-,
Z .1~
T
U .011
| .. ~ q~ .
.04 Z
1
5_< .oz ,d
mr U
0
!l I
- .02 . . . . I i I Z
<
A
-~
ei
0700 OgO0 II~ ' 1300 0700 OlO0 IlO0 I~O0 e
o
T I M E OF D A Y ( h i T I M E OF D A Y ( I I )
Z
Figure 5. Arterial plasma ~-amino nitrogen (c~-NH2 ~ q ~ ~. .
N), arteriovenous concentration difference and c~-NH2
N uptake in steers at each sampling time, Sampling
began at 0700 and continued for 6 h at 30-min inter-
vals. The arrow indicates time of feeding. A: Samples
taken on d 1 of control ( o - - e ) or ctenbuterol (o--o)
treatment. Average SEM for control and clenbuterol
treatments, respectively, was: ~-NH~ N (mM) .20 and
.04; arteriovenous a-NH~ N (raM) .03 and .02; ~-NH~
N uptake (mmol/min) .12 and .12. B: Samples taken
on d 9 of control ( o - - ~ ) or clenbuterol (o--o) treat- I
ment. Average SEM for control and clenbuterol treat-
ments, respectively, was: ~-NHz N (raM) .21 and .12;
arteriovenous e~-NH~ N (raM) .03 and .02; ce-NH2 N
uptake (mmol/min) .13 and .11. <
X "x~
I
increased w i t h c l e n b u t e r o l c o m p a r e d w i t h con-
t r o l feeding. By d 9, c l e n b u t e r o l f e e d i n g caused
a s u s t a i n e d increase in u p t a k e o f 0 2 (figure 2 b ;
t a b l e 4), increased u p t a k e of a-NH2 N (figure e~
ably indicates that the drug is absorbed in the crease cardiac output 33% if stroke volume
proximal sections of the gastrointestinal tract. were unchanged. The change in distribution of
Clenbuterol has been described mainly as a cardiac output may be due to stimulation of
132-adrenergic agonist because of effects on arterial fl-adrenergic receptors located in skele-
bronchodilation; however, the marked tachy- tal muscle (Smith and Hamlin, 1977) and(or)
cardia caused by administration of the com- may be a result of increased tissue metabolism.
pound to steers suggests fll-adrenergic activity, The acute changes in blood concentration of
as defined by Lands et al. (1967). By d 9, metabolites (figures l a to 5a) reflect the extent
chronic tachycardia in response to clenbuterol of metabolic perturbation induced by clenbuter-
feeding was still present (figure lb). Williams et ol. Of the metabolites exhibiting changes in
al. (1986) reported tachycardia in young bull concentration, those associated with carbo-
calves fed clenbuterol at 20 lag/kg BW; however, hydrate and lipid metabolism (glucose, L-lac-
adaptation, indicated by a return o f heart rate tate, /~-hydroxybutyrate and N E F A ) increased,
to pretreatment values, occurred b y d 3 of whereas those related to N metabolism (0~-NH2
treatment. Acute tachycardia in response to N, NH3) decreased. Combined information on
clenbuterol (Herbert et al., 1985) or cimaterol changes in concentration and uptake or release
(Beermann et al., 1986a) was observed in sheep. elucidates whether entry or exit from the plas-
An acute increase in blood flow to the hind- ma pool is altered to produce nonsteady-state
quarters in response to cimaterol was observed conditions. Our data suggest that increased con-
also in sheep (Beermann et al., 1986a). Increased centration of glucose reflects increased entry
PCV in response to clenbuterol on d 1 (table 2) through an increase in liver gluconeogenesis
may reflect splenic contraction, generally and(or) glycogenolysis. Similarly, increased
described as an a-adrenergic mediated event entry of lactate through an increase in peri-
(Ignarro and Titus, 1968). pheral glycolysis and of N E F A through in-
Isoproterenol, a nonselective 13-adrenergic creased lipolysis are consistent with observed
agonist, acutely increased both heart rate and changes. Thornton et al. (1985) demonstrated a
stroke volume to increase cardiac output in the direct acute effect of clenbuterol added to in
guinea pig (Van de Walle and Martin, 1985). vitro incubations of sheep adipose tissue to
Total peripheral resistance was decreased. In increase lipid mobilization. The acute changes
response to isoproterenol, absolute blood flow in carbohydrate and lipid metabolites were
to the carcass and portal-drained visceral (PDV) similar to ~ and 13-adrenergic-mediated metabo-
tissues was increased, as was the proportion of lic effects previously reported in sheep (Bassett,
cardiac output distributed to the carcass of the 1970).
guinea pigs, suggesting a selective vasodilation Decreased concentration of~-NH2 N coupled
in response to a /3-adrenergic agonist, in addi- with decreased peripheral uptake on d 1 indi-
tion to increased overall flow. We have observed cate a decrease in entry rate, most likely through
increased blood flow to both the hindquarters decreased protein degradation. Short-term
and PDV in steers following initial ingestion of studies using skeletal muscle from rats demon-
2 mg clenbuterol, and the increase in blood flow strated a/3-adrenergic-mediated decrease in pro-
to the hindquarters was greater than that to the tein degradation (Li and Jefferson, 1977) and
PDV (unpublished observations). Administra- amino acid release (Garber et al., 1976). The
tion of clenbuterol to horses acutely increased time course of changes in a-NH~ N concentra-
cardiac output by 44%, whereas cardiac output tion suggesting decreased protein degradation is
was increased only 15% after 8 d of clenbuterol consistent with the observed increase in N reten-
treatment (16/ag/kg BW; Claussen, 1981). These tion on d 1 of clenbuterol (Herbert et al., 1985)
cited studies support the concept that the ini- or cimaterot (Beerman et al., 1986b) infusion in
tial increase in blood flow to the hindquarters lambs nourished by intragastric infusion. Epine-
was caused by increased cardiac output, as well phrine (a and j3-adrenergic agonist) had no effect
as selective vasodilation in response to clenbu- on in vitro protein synthesis in diaphragm mus-
terol. Further, these studies suggest that the cle from rats (Nutting, 1982).
large (47%) chronic increase in blood flow to Chronic effects of clenbuterol on metabolite
the hindquarters on d 9 of clenbuterol feed- concentrations contrasted with acute effects
ing (table 2) partly reflects a change in distri- observed on d 1. Lack of acute metabolic
bution of cardiac output. The increased heart changes on d 9 demonstrates adaptation to the
rate on d 9 o f clenbuterol feeding would in- drug and a change in tissue sensitivity to clen-
buterol. The change in sensitivity could be re- suggests increased glycolysis with clenbuterol
lated to receptor of post-receptor mediated feeding. Mobilization and increased oxidation
events. Concentrations of V F A decreased and of NEFA inhibits glucose oxidation at pyruvate
concentrations of N E F A increased chronically, dehydrogenase, with consequent increased
whereas concentration of variables associated release of lactate (Newsholme and Leech,
with carbohydrate and N metabolism were un- 1983). The fact that glucose uptake was not
changed. Previous studies demonstrated a posi- altered on d 9 of clenbuterol feeding, but that
tive correlation between whole body irreversi- lactate release was (table 4), exemplifies this
ble loss or oxidation and b l o o d concentrations metabolic shift. On the average, glucose uptake
of acetate (Annison et al., 1967; Pethick et al., corrected for lactate release 11 accounted for
1981) or NEFA (Lear and Ford, 1966; Pethick 39.8 and 29.1%, maximum, of 02 uptake for
et al., 1983; Eisemann et ai., 1986); however, control and clenbuterol treatments, respec-
this association did not hold for leucine (Else- tively, on d 9. The potential contribution of
mann et al., 1986). Decreased entry of acetate f3-hydroxybutyrate to O212 use was constant at
from the portal-drained viscera or decreased 19.8 and 21.9%, respectively. Because acetate is
endogenous hepatic production may be re- used for both oxidation and lipid synthesis,
sponsible for decreased entry. Decreased AV potential contribution to 02 consumption was
concentration difference (table 3) and a trend not calculated.
toward decreased acetate uptake in the hind- By d 9 of treatment, hindquarters uptake of
quarters (table 4) supports decreased acetate a-NHR N was increased, suggesting an increase
use (exit) for oxidation and(or) lipogenesis on in protein deposition in response to chronic
a whole-body basis. Chronically elevated arterial clenbuterol treatment. Chronic administration
N E F A concentrations suggest continued lipid of clenbuterol to rats resulted in either an in-
mobilization and oxidation with chronic clen- crease (Emery et al., 1984) or no change (Reeds
buterol treatment. N E F A could also be directly et al., 1986) in fractional synthesis rate of mus-
oxidized, from hydrolysis of triacylglycerol in cle protein. Also, the fractional synthesis rate
muscle tissues, without entering the plasma of muscle protein was not altered in sheep with
pool. Consistent with the tendency toward de- chronic clenbuterol administration (Bohorov et
creased acetate uptake, there was a trend for al., 1987). Emery et al. (1984) administered 2
increased lipid mobilization from the hind- mg-(kg BW~ - 1 subcutaneously, whereas
quarters; however, other body depots may Reeds et al. (1986) fed clenbuterol at approxi-
make a larger contribution to total lipid mobili- mately 160 to 5 6 0 / l g . ( k g BW-d) - 1 . Bohorov
zation. et al. (1987) included clenbuterol in the feed to
For O~ and metabolites monitored, mean effect a dose of 17 m g . ( h d . d ) - 1 for sheep from
AV differences and extraction ratios changed in 27 to 40 kg BW. Dose or method of administra-
similar directions with both acute and chronic tion may contribute to the variation in observed
clenbuterot feeding (table 3). Increased use of effects.
02 in the hindquarters (table 4), indicating in- Oxygen consumption by the hindquarters
creased oxidative metabolism, could result from was increased 3.06 mmol/min (4.41 mol/d) by
increased catabolism of N E F A or glucose. If chronic clenbuterol treatment. Assuming tight-
glucose is metabolized to lactate, rather than ly coupled mitochondria (i.e., 1 mol O2 con-
completely oxidized to H 2 0 and COs, this sumed equals 6 mol ATP formed), increased 02
would not be accompanied by an increase in 02 consumed is equivalent to the formation of a
consumption in the hindquarters because ATP maximum of 26.5 tool of ATP. The increment
production would be at the substrate level in a-NH 2 N uptake was .216 mol/d or 148.5 g
rather than coupled to electron transport. The crude protein (assuming 16% N and average
large release of lactate on d 1 of clenbuterol molecular weight 110 g/mol amino acid). If 5
feeding and continued release on d 9 (table 4) mol ATP are the minimum required/peptide
b o n d formed (equivalent to 1 tool amino acid
or 110 g protein; Millward and Garlick, 1976),
then 6.75 tool of ATP or 25.6% of the increased
11Calculated stoichiometrieally based on: 1 glucose ATP would be needed if the 148.5 g increment
+ 6 0 z ~ 6 CO2 + 6 HzO. Each mole lactate release is in crude protein resulted from protein synthesis.
equivalent to .5 mole glucose.
12Calculated stoichiometrically based on: 1 3- Increased NEFA oxidation could provide the
hydroxybutyric acid + 4.5 0 2 ~ 4 CO 2 + 4 HzO. needed ATP. Increased substrate cycling (News-
Soar. 1987. The effect of the /3-2-adrenergic NRC. 1984. Nutrient Requirements of Beef Cattle.
agonist clenbuterol or implantation with oestra- Sixth Revised Ed. National A c a d e m y of Sciences-
diol plus trenbolone acetate on protein m e t a b o - National Research Council. Washington, DC.
lism in wether Iambs. Br. J. Nutr. 57:99. Nutting, D. F. 1982. Anabolic effects of catechola-
Claussen, H. 1981. B e s t i m m u n g des Herzminuten- mines in diaphragm muscle f r o m h y p o p h y s e c t o -
volumens mit der T h e r m o d i l u t i o n s m e t h o d e bei mized rats. Endocrinology 11 ~307.
Pferden vor u n d nach Verabreichung yon Clenbu- Pethick, D. W., D. B. Lindsay, P. J. Barker and A. J.
terol oder Carazolol. Inaugural Dissertation, Han- Northrop. 1981. Acetate supply and utilization
nover, Tierarztliche. Hochschule No. 1 - 7 . by t h e tissues o f sheep in vivo. Br. J. Nutr. 46:97.
Dalrymple, R. H., P. K. Baker and C. A. Ricks. 1984. Pethick, D. W., D. B. Lindsay, P. J. Barker and A. J.
Repartitioning agents to improve performance Northrop. 1983. T h e metabolism of circulating
and b o d y composition. Proc. Georgia Nutr. Conf. non-esterified f a t t y acids by t h e whole animal,
pp 111--118. hind-limb muscle and uterus of pregnant ewes.
Eisemann, J. H., A. C. H a m m o n d , D. E. Bauman, P. J. Br. J. Nutr. 49:129.
Reynolds, S- N. McCutcheon, H. F. Tyrrell and Prior, R. L. 1978. Effect o f level of feed intake on
G. L. Haaland. 1986. Effect of bovine growth lactate and acetate metabolism and lipogenesis in
h o r m o n e administration on metabolism of grow- vivo in sheep. J. Nutr. 108:926.
ing Hereford heifers: Protein and lipid metabo- Prior, R. L., G. B. H u n t i n g t o n and P. J. Reynolds.
lism and plasma concentrations of metabolites 1984. Role of insulin and glucose on metabolite
and hormones. J. Nutr. 116:2504. uptake by t h e hind half of beef steers. J. Anim.
Emery, P. W., N. J. Rothwell, M. J. Stock and P. D. Sci. 58:1446.
Winter. 1984. Chronic effects of ~32-adrenergic Reeds, P. J., S. M. Hay, P. M. Dorwood and R. M.
agonists on b o d y composition and protein syn- Palmer. 1986. Stimulation o f muscle growth b y
thesis in t h e rat. Biosci. Rep. 4:83. clenbuterol: Lack of effect on muscle protein
Garber, A. J., I. E. Karl and D. M. Kipnis. 1976. Ala- biosynthesis. Br. J. Nutr. 56:249.
nine and glutamine synthesis and release from Reynolds, P. J., G. B. H u n t i n g t o n and C. K. Reynolds.
skeletal muscle. 4. ~3-adrenergic inhibition o f 1986. Determination o f volatile fatty acids,
amino acid release. J. Biol. Chem. 251:851. lactate, and 3 - h y d r o x y b u t y r a t e in blood b y ion
Hanrahan, J. P., J. F. Quirke, W. Bomann, P. Allen, J. exchange cleanup and gas chromatography. J.
McEwan, J. Fitzsimons, J. Kotzian and J. F. Anim. Sci. 63 (Suppl. 1):424.
Roche. 1986. ~3-agonists and their effects on Ricks, C. A., R. H. Dalrymple, P. K. Baker and D. L.
growth and carcass quality. In: W. Haresign and Ingle. 1984. Use of a 3-agonist to alter fat and
D. J. Cole (Ed.) Recent Advances in Animal muscle deposition in steers. J. Anita. Sci. 59:
Nutrition. pp 125--138. Butterworth, L o n d o n . 1247.
Herbert, F., F. D. DeB Hovell and P. J. Reeds. 1985. Smith, C. R. and R. L. Hamlin. 1977. Regulation of
Some preliminary observations on the immediate t h e heart and blood vessels. In: M. J. Swenson
effects of clenbuterol on heart rate, b o d y t e m - (Ed.) Dukes' Physiology of Domestic Animals. pp
perature and nitrogen retention in lambs wholly 1 0 2 - 1 2 0 . Cornell University Press, Ithaca, New
nourished b y intragastric infusion. Proc. Nutr. York.
Soc. 44:150A. T h o r n t o n , R. F., R. K. T u m e , G. Payne, T. W. Larsen,
Ignarro, L. J. and E. Titus. 1968. The presence of G. W. J o h n s o n and M. A. Hohenhaus. 1985. The
antagonistically acting alpha and beta adrenergic influence of t h e 32-adrenergic agonist, clenbu-
receptors in t h e m o u s e spleen. J. Pharmacol. Exp. terol, On lipid metabolism and carcass composi-
Ther. 160:72. tion of sheep. Proc. N. Z. Soc. Anim. Prod.
Lands, A. M., A. Arnold, J. P. McAuliff, F. P. L u d u e n a 45:97.
and T. G. Brown. 1967. Differentiation o f Van de Walle, A.F.G.M. and C. B. Martin. 1985. Effect
receptor s y s t e m s activated b y s y m p a t h o m i m e t i c of isoproterenol on uterine blood flow and
amines. Nature 214:597. cardiac o u t p u t distribution in pregnant guinea
Lear, W.M.F. and E.J.H. Ford. 1966. utilization of pigs. A m . J. Obstet. Gynecol. 152:1058.
free fatty acids by starved and pregnant sheep. Whitehurst, G. B., D. C. Beitz, M. A. Pothoven, W. R.
Biochem. J. 101:317. Ellison and M. H. Crurnp. 1978. Lactate as a
Li, J. B. and L. S. Jefferson. 1977. Effect o f isopro- precursor of f a t t y acids in bovine adipose tissue.
terenol on amino acid levels and protein turnover J. Nutr. 108:1806.
in skeletal muscle. A m . J. Physiol. 232:E243. Williams, P.E.V., L. Pagliani and G. M. Innes. 1986.
Millward, D. J. and P. J. Garlick. 1976. The energy The effect of a 3-agonist (clenbuterol) on t h e
cost of growth. Proc. Nutr. Soc. 35.339. heart rate, nitrogen balance and some carcass
Newsholme, E. A. and B. Crabtree. 1976. Substrate characteristics of veal calves. Livestock Prod. Sci.
cycles in metabolic regulation and in heat genera- 15:289.
tion. Biochem. Soc. Symp. 41:61. Williamson, D. H. and J. Mellanby. 1974. D-(-)-3-
Newsholme, E. A., and A. R. Leech. 1983. Biochem- h y d r o x y b u t y r a t e . In: H. U. Bergmeyer (Ed.)
istry for the Medical Sciences. J o h n Wiley and Methods of Enzymatic Analysis. pp 1836--1839.
Sons, New York. Academic Press, New York.