Fisheries Science 61(2), 277-281 (1995)

Metabolic Response to Dietary Carbohydrate to Protein Ratios in Carp

Sadao Shimeno, Driss Kheyyali,•õ and Takafumi Shikata

Laboratory of Fish Nutrition, Faculty of Agriculture, Kochi University, Monobe, Nankoku, Kochi 783, Japan

(Received July 6, 1994)

Common carp Cyprinus carpio were fed at a constant feeding rate for 30 days on isoenergetic diets
containing different levels of carbohydrate (4-58%) and protein (65-27%), and activities of several
hepatopancreatic enzymes together with growth performance were determined. Dietary inclusion of
adequate levels of carbohydrate improved the growth and feed conversion and exerted a protein-spar
ing effect. With increasing dietary carbohydrate levels and decreasing protein levels, the activities of
hepatopancreatic glucosephosphate isomerase, glucose-6-phosphate and phosphogluconate de
hydrogenases, and malic enzyme as well as hepatopancreatic glycogen and serum triglyceride contents
increased, while those of glucose-6-phosphatase, arginase, GOT, and GPT together with serum free
amino acid concentrations decreased. Therefore, high correlation coefficients were obtained between
dietary carbohydrate levels and these biochemical parameter concentrations. The results suggest that
dietary carbohydrate stimulates glycolysis and lipogenesis and depresses gluconeogenesis and amino
acid degradation in the hepatopancreas. This comprehensive regulation of nutrient metabolism may ac
count for the protein-sparing effect exerted by dietary carbohydrate.
Key words: carp, metabolic response, hepatic enzyme, dietary composition, carbohydrate,
ptotein

Carbohydrate is an important energy source for fish as it ing amount of each experimental diet, the feeding rate was
is for mammals, and it is well known that the significance found to be the lowest in the fish fed on the highest protein
of the nutrient differs depending on fish species.1-3) diet. However, the authors previously reported that lo
However, limited information is available on the metabol wered feeding rates depressed hepatopancreatic glycolysis
ic response to dietary carbohydrate in fishes, and its and lipogenesis in the fish.7,8) Moreover, Murai et al.)8 de
biochemical foundation remains unclear.4) The authors scribed that feeding frequency affected the growth and
reported in the previous papers) that herbivorous fish tila body composition in carp. Therefore in the experiment on

pia could adapt to high carbohydrate diets such as diets metabolic response to dietary composition, carp were fed
containing up to 50% starch by increasing its assimilation at constant daily feeding rate (2.3%) and twice a day at
and preservation, while carnivorous fish such as yellowtail 9:00 and 16:00 on one of the 5 experimental diets described
could not adapt to these high carbohydrate diets, resulting later, for 30 days from July 22. The water temperature dur
in metabolic disorders such as glucose intolerance and ing the experimental period ranged between 16.5-19.5•Ž.
lowered glycolysis. 1,3,6)
On the other hand, nutrient metabolisms, especially Experimental Diet
glycolysis and lipogenesis, were known to be affected by Five diets were used in the experiment (Table 1). Name
not only dietary composition but also the feeding rate for ly, white fish meal and wheat gluten were used as a protein
fishes.7,8) In the present study, carp were fed at a constant source, and the mixture of ƒ¿-potato starch and dextrin
feeding rate for 30 days on isoenergetic diets containing (1:1) as a carbohydrate source. Powdered pollack liver oil
different levels of protein and carbohydrate, and the activi was added to adjust dietary lipid levels and to supply their
ties of hepatopancreatic enzymes and body composition as essential fatty acids. Dietary ingredients were mixed thor
well as the growth and feed conversion were determined. oughly and pelleted using a laboratory pellet mill. From
diets I to 5, crude sugar level was increased from 3.5 to
Materials and Methods 57.5%, while the protein level was decreased from 64.6 to
26.5%. The energy levels ranged from 3650 to 3530 kcal/
Fish and Feeding Procedure kg dry diet.
Carp Cyprinus carpio of 49.5 g in average body weight
were used for the experiment. The fish were randamly Assay of Serum and Body Components
divided into 5 groups of 30 individuals each and reared in At the end of the experiment, about ten fish from each
8001 flow through fiber reinforced plastic aquaria with a group were randomly selected at 17 h after the last meal to
flow rate of approximately 21/min in Freshwater Feeding determine the body and serum composition and
Room, Kochi University. In a preliminary test on the feed hepatopancreatic enzymes. General composition of the

* Regulation of Carbohydrate Metabolism in Fish -XX‡V.

•õ Present address: Department of Animal Physiology. Hassan ‡U University, B.P.6202 Rabat, Morocco.
278 Shimeno et a!.

Table 1. Composition of diets for the experiment Table 2. Performance of fish fed for 30 days on test diets

*1 100 •~ Dry diet fed (g)/ 100 g body weight/day

.
*2 100 •~ Weight gain (g)/dry diet fed (g)
.
*3 Weight gain (g)/protein fed (g)
.
*4 100 •~ Weight gain (g)/energy fed (kcal)
.

*1 a -Potato starch: dextrin=1:1 .

*2 Halver (1957)
.14)
*3 Ogino et al . (1979).13)
*4 Calculated energy .3,7,11) Table 3. Proximate composition of the fish at the end of the experi
ment (%)

whole body, hepatopancreas, and muscle was determined

by usual methods.5-8) The blood of 5 fish was individually
drawn from the caudal vein with a syringe, and the concen
trations of serum protein, free amino acid, triglyceride,
phospholipid, cholesterol, free fatty acid, and glucose
were determined by the methods described previously.5,7)

Enzyme Assay
Immediately after killing, the hepatopancreas of the 5
fish was removed, weighed, and individually frozen in liq
uid nitrogen, and stored at -80•Ž for enzyme assay.
Then, it was homogenized with 9 volumes of cold water
for 1 min in a Potter-Elvehjem's glass homogenizer. After
centrifuging at 5000 rpm for 10 min at 0-5•Ž, the resulting
supernatant was used for assaying the following enzymes:

glucosephosphate isomerase (PGI, EC 5.3.1.9); glucose-6

phosphatase (G6Pase, EC 3.1.3.9); glucose-6-phosphate
-
dehydrogenase (G6PDH, EC 1.1.1.49); phosphogluconate * Pooled sample of 3 (whole body and muscl
e) and 7 fish (hepatopancreas).
dehydrogenase (PGDH, EC 1.1.1.44); malate de
hydrogenase (decarboxylating) (NADP-MDH. EC
1.1.1.40); isocitrate dehydrogenase (NADP-ICDH, EC
1.1.1.42); aspartate aminotransferase (GOT,EC 2.6.1.1); drate resulted in a substantial decrease in weight gain. A
alanine aminotransferase (GPT, EC 2.6.1.2); arginase (EC similar tendency among groups was found for feed efficien
3.5.3.1). The enzyme activities were determined by the cy and energy efficiency, although the efficiencies were
methods described previously,5-8) and they were expressed generally low depending on low temperatures and restrict
asp mol of a substrate or coenzyme converted per min per ed feeding rate. Protein efficiency ratio improved as dieta
100 g body weight. ry carbohydrate level was increased from 4 to 48% (groups
Statistical analyses for significance (p<0.05) of the data I to 4), while no more improvement was observed in the
on serum components and enzymes were carried out ac group 5 fed on the diet containing 27% protein and 58%
cording to Duncan's multiple range test.10) carbohydrate.

Results Proximate Composition

As is evident from Table 3, increasing the dietary carbo
Growth Performance hydrate levels resulted in an increase of glycogen or total
The average daily feeding rate was equal among groups sugar content in the hepatopancreas, whole body, and mus
(2.3-2.4%) fed on diets containing different carbohydrate cle. Fat content in the whole body also increased with in
to protein ratios (Table 2), and its ratios affected the creases in carbohydrate levels, but not in the hepatopan
growth and feed conversion. The average weight gain was creas and muscle, suggesting that fat was deposited mainly
highest in groups 3 and 4, fed diets containing 32 and 48% in the visceral cavity. The other body components were
carbohydrate, and more and less inclusion of carbohy not so markedly affected by dietary composition, though
 Metabolic Response to Dietary Carbohydrate in Carp 279

Table 4. Effect of dietary carbohydrate to protein ratios on the concentrations of serum components

*1 Correlation coefficient between serum component concentrations and carbohydrate contents in diets .
*2 Mean •}SD of 5 fish . Values with the same superscripts are not significantly different (p<0.05).

Table 5. Effect of dietary carbohydrate to protein ratios on the activities of hepatopancreatic enzymes
(ƒÊ mol/min/100 g body weight)

*1 Correlation coefficient between enzyme activities and carbohydrate contents in diets.

*2 Mean•}SD of 5 fish
. Values with the same superscripts are not significantly different (p<0.05).

protein content gradualy decreased
carbohydrate levels.
Serum Component
Table 4 shows the effect of dietary carbohydrate to pro
tein ratios on serum components. The concentrations of
triglyceride, phospholipid, and free fatty acid increased
with increases in dietary carbohydrate levels, and they
were highest in the group 5 fed on the highest-carbohy
drate and lowest-protein diet. Therefore, these lipid con
centrations positively correlated with the carbohydrate lev
els in diets (r=0.93-0.99), and significant differences were
recognized in these concentrations among groups.
Although the free amino acid concentration was sig
nificantly higher in the group 1 fed the highest protein diet
than the other groups, the glucose concentration did not
show any trend relating to the dietary composition.
Hepatopancreatic Enzyme
Of the enzymes investigated, NADP-MDH was most
responsible for a difference in dietary carbohydrate to pro
tein ratios (Table 5). Namely, the enzyme activity was
lowest in group 1, then increased gradually and was the
highest in group 5 fed the diet containing 27% protein and
58% carbohydrate. There were significant differences in
the activity between all the two groups tested. A similar
tendency among groups was detected for activities of the
other lipogenic enzymes (G6PDH. PGDH, and NADP-
ICDH) together with glycolytic enzyme PGI, but their
response to dietary carbohydrate was less pronounced.
with increasing dietary These enzyme activities positively correlated with dietary
carbohydrate levels (r=0.90-0.98), and significant differ
ences were found in the activities among groups.
On the other hand, the activities of gluconeogenic en
zyme (G6Pase) and amino acid-degrading enzymes (GOT,
GPT, and arginase) were highest in group 1, then
decreased with increasing carbohydrate levels in diet, and
were lowest in group 5. Therefore, there were negative cor
relations between each enzyme activity and dietary carbo
hydrate levels (r=-0.92~-0.97), and significant differ
ences were detected between groups 1.2 and groups 4.5.
Discussion
In order to clarify the regulatory mechanism of nutrient
metabolism in the fish fed high carbohydrate diets, the ra
tios of serum and liver constituent contents and hepatic en
zyme activities in the group 5 fed on the highest carbohy
drate diet to those in the control group fed on the lowest
carbohydrate diet were calculated and are shown in Fig. 1.
In the carbohydrate-fed group as compared with the con
trol group, the activities of glycolytic and lipogenic en
zymes PGI, G6PDH, PGDH, and NADP-MDH as well as
body glycogen and lipid contents were significantly higher,
while those of gluconeogenic and amino acid-degrading en
zymes G6Pase, GOT, GPT, and arginase were significant
ly lower. Furthermore, since high correlation coefficients
were obtained between the dietary carbohydrate levels and
enzyme activities or serum component concentrations
(Tables 4 and 5), a similar tendency was also found for all
 280 Shimeno et al .

to 27:58 (diet 5) or decreased down to 65:4 (diet 1). There

is much information indicating that moderate inclusion of
carbohydrate into the fish diet ameliorates the growth Per
formance.1,3,11)These growth performances together with
the metabolic response described above, indicate that carp
tolerate a comparatively high level of dietary carbohy
drate, and that the fish have a high ability to utilize both
protein and carbohydrate in their diet.
In order to estimate the protein-sparing effect of carbo
hydrate in diet, the amount of dietary protein required for
10 g weight gain was calculated and is shown in Table 2 .
Fig. 1. Metabolic adaptation to dietary carbohydrate in carp. Values The amount of the highest protein diet in group 1 was
represent the ratios of the activity, content, and concentration in fish 18.8 g, while those of the four groups fed diets containing
fed on the highest carbohydrate diet 5, to those in fish fed on the car lower protein and higher carbohydrate were 15.8, 10.6,
bohydrate-free diet 1. Arrows in boldface and dashed lines indicate 8.3, and 8.0 g, respectively. Taking the highest protein
significant differences between groups.
group as a control, the protein amount required for 10g
weight gain could be spared under the rearing conditions
by approximately 14, 43, 55, and 57% in groups 2, 3, 4,
the parameters examined between the control group and and 5, respectively. Namely, when more carbohydrate was
groups 3 or 4 fed on diets containing less amounts of carbo supplemented, the more dietary protein was spared in the
hydrate. These findings might indicate that feeding carbo fish as it was for tilapia and yellowtail.3.51
hydrate accelerates their glycolysis and lipogenesis and As is mentioned above, carp have a high carbohydrate
depresses gluconeogenesis and amino acid degradation in utilizability, so that adding a large amount of carbohy
the hepatopancreas. drate into the diet can spare a lot of dietary protein
Similar regulatory pattern to adequate levels of carbohy without retarding the growth. As to the regulatory mecha
drate in diet was observed in carp, tilapia, channel catfish, nism of protein-sparing effect by dietary carbohydrate, it
and yellowtail.3,5.6,11-13) However, feeding a high carbohy is suggested that feeding more carbohydrate depressed ami
drate diet to yellowtail resulted in a different pattern such no acid catabolism and gluconeogenesis as shown in Fig .
as lowered glycolysis and lipogenesis, showing glucose in 1, so that less amino acid could be used for energy and glu
tolerance and growth retardatioon.1,3,6) These findings lead cose production.
to the conclusion that there is no substantial difference in
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