Journal of Thermal Biology 37 (2012) 686695

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Journal of Thermal Biology

journal homepage: www.elsevier.com/locate/jtherbio

Growth and IGF-I response of juvenile Nile tilapia (Oreochromis niloticus)

to changes in water temperature and dietary protein level
J. Qiang a,b, H. Yang a,b, H. Wang c, M.D. Kpundeh a, P. Xu a,b,n
a

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China

Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences,
Wuxi 214081, Jiangsu, China
c
Fisheries College, Guangdong Ocean University, Zhanjiang 524025, China
b

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 16 June 2012
Accepted 31 July 2012
Available online 23 August 2012

Water temperature and dietary protein level play an important role in inuencing the growth and insulinlike growth factor I (IGF-I) in Nile tilapia juveniles. The combined effect of temperature (2034 1C) and
dietary protein level (2550%) on the specic growth rate (SGR), feed efciency (FE), serum IGF-I level and
hepatic IGF-I mRNA level was examined under laboratory conditions by employing central composite
design and response surface method. Results showed that the linear effects of temperature and dietary
protein level on the SGR, FE, serum IGF-I and hepatic IGF-I mRNA level were signicant (Po0.05); the
quadratic effects of temperature and dietary protein level on the FE and serum IGF-I were signicant
(Po0.05). The interaction of temperature and dietary protein level on the FE, serum IGF-I and hepatic IGF-I
mRNA level all proved signicant (Po0.05). The optimal temperature/dietary protein level combination
was determined, i.e., 29.9 1C/40.3%, at which the greatest SGR (2.748%/d) and FE (0.775) were simultaneously arrived. Both SGR and FE were linearly correlated with serum IGF-I or hepatic IGF-I mRNA level.
These results suggested that optimum combination of temperature and dietary protein level would
enhance tilapia growth efciency and IGF-I would regulate growth and FE.
& 2012 Elsevier Ltd. All rights reserved.

Keywords:
Nile tilapia (Oreochromis niloticus)
Water temperature
Dietary protein level
Growth
Insulin-like growth factor I
Response surface

1. Introduction
The quality and high yield of commercial shes are intimately
related to feed quality, feeding regime and the rearing environment.
Typically, protein makes up a larger proportion of the feed, and at
the same time is also one of the most important dietary nutrients
(Singh et al., 2006). The intake of proteins is advantageous for tissue
renewal/repair and metabolism in sh (Alvarez-Gonzalez et al.,
2001). When the level of dietary proteins is greater than 40%, the
growth of tilapia juveniles will be promoted (Al Hafedh, 1999);
whereas deciency of dietary proteins can affect tilapia growth
and survival (Al Hafedh, 1999), gonadal vitelline development
(Gunasekera and Lam, 1997), and fertilization (El-Sayed and
Kawanna, 2008). Therefore, higher levels of protein is typically
needed in sh feed. For instance, a dietary protein level of 50% is
required by juvenile starry ounder, Platichthys stellatus (Lee, 2006);
3845% by juvenile black sea bream, Sparus macrocephalus (Zhang
et al., 2010); and 50% by juvenile silver pomfret, Pampus argenteus
(Hossain et al., 2010).

n
Corresponding author at: Wuxi Fisheries College, Nanjing Agricultural
University, Wuxi 214081, China. Tel./fax: 86 510 85557959.
E-mail address: Xup@ffrc.cn (P. Xu).

0306-4565/$ - see front matter & 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jtherbio.2012.07.009

The requirement of dietary proteins for aquatic animals is closely

associated with water temperature (Peres and Oliva-Teles, 1999;
Singh et al., 2009), salinity (Perez-Velazquez et al., 2007), size
(Abdel-Tawwab et al., 2010), rearing density (Omar et al., 1997),
dietary lipid level (Carmona-Osalde et al., 2005), dietary energy
(Pirozzi et al., 2010), sources of protein (Garcia and Villarroel, 2009;
Zheng et al., 2011), and so forth. Temperature is a crucial factor in
aquaculture; it has direct impacts on sh biochemical reactions
(Streit Jr et al., 2010). Singh et al. (2008) found that when water
temperature ranged between 28 1C and 32 1C, the dietary protein
requirement for mrigal Cirrhinus mrigala fry was 36%, and the
protein utilization rate was higher at dietary protein level of 28%.
When water temperatures were 15 1C and 20 1C, optimal protein
level of sea bass juveniles Dicentrarchus labrax was 50%, but the daily
protein requirement and food intake were markedly higher at 20 1C
than at 15 1C (Hidalgo and Alliot, 1988). Increasing water temperature quickens the sh energy metabolism within certain range
(Likongwe et al., 1996). There have been many reports on the
inuence of water temperature or dietary protein level upon the
growth and feed utilization in tilapia (Abdel-Tawwab et al., 2010; Al
Hafedh, 1999; Azaza et al., 2008; Likongwe et al., 1996). Using
factorial experimental design, Musuka et al. (2009), examined the
effect of dietary protein requirement and water temperature on the
growth performance of Tilapia rendalli; this is the only report in

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

terms of the inuence of dietary protein level at differing water

temperatures in tilapia so far.
Neuroendocrine factors play a role in regulating and controlling sh growth and energy metabolism (Beckman et al., 2004).
Growth hormone (GH), insulin-like growth factor (IGF-I) and
insulin, are three important neuroendocrine factors that can
contribute greatly to growth and metabolism (Planas et al.,
os et al., 1999). GH level can directly affect the secretion
2000; Ban
of IGF-I in liver. The production and secretion of these hormones
are directly or indirectly related to some exogenous factors, such
as nutritional status (Cameron et al., 2007; Riley et al., 2009; Enes
et al., 2010; Jiang et al., 2010), feeding frequency (Petersona et al.,
2009), water temperature (Imsland et al., 2007), salinity (Taylor
et al., 2008), photoperiod (Cruz and Brown, 2009) and season
(Davis and McEntire, 2006). For example, increased water temperature led to an increased plasma IGF-I levels in Chinook
salmon Oncorhynchus tshawytscha (Beckman et al., 1998), turbot
Scophthalmus maximus (Imsland et al., 2007), sunshine bass
(Morone chrysops  Morone saxatilis) (Davis and McEntire,
2011); as well as increased hepatic IGF-I mRNA level in Nile
tilapia, Oreochromis niloticus (Vera Cruz et al., 2006). Dietary
protein level is also closely correlated with the level of GH and
IGF-I (Perez-Sanchez et al., 1995; Dyer et al., 2004). No reports
have been found thus far, however, on the interactive effect
between water temperature and dietary protein level on growth,
feed efciency, serum IGF-I and hepatic IGF-I mRNA level in Nile
tilapia.
In order to gain a better understanding of growth and IGF-I
response in the Nile tilapia under different temperatures and dietary
protein levels, this study was conducted and aimed at examining: (i)
the combined effect of water temperature and dietary protein level
on growth, feed efciency, serum IGF-I and hepatic IGF-I mRNA level
in Nile tilapia juveniles under laboratory conditions; (ii) the quadratic effect of the above two factors and predictive model equations
of growth, feed efciency, serum IGF-I and hepatic IGF-I mRNA level
towards water temperature and dietary protein level; (iii) the
relationship of growth and feed efciency to serum IGF-I and
hepatic IGF-I mRNA level. This study was conducted to provide
scientic basis for growth regulation in Nile tilapia.

2. Materials and methods

2.1. Experimental sh
Healthy Nile tilapia juveniles, of the 16th generation of the
Genetically Improved Farm Tilapia (GIFT) strain, were obtained as

687

experimental sh from Yixing, the experimental base of Freshwater Fisheries Research Center of Chinese Academy of Fishery
Sciences. Prior to the experiment, these sh were acclimatized
for 10 days in an indoor cement pool with water temperature
27 1C70.3, under natural photoperiod and continuous aeration
using recirculating water. The sh were trained to accept
submerged feed (crude protein 37.5%, fat 8.0%).
2.2. Experimental design
In order to examine the relationship between the responses,
which were the specic growth rate (SGR), feed efciency (FE),
serum IGF-I level and hepatic IGF-I mRNA level, and the factors of
interest, which were temperature and dietary protein level,
central composite design was used in this experiment. Temperature ranged from 20 1C to 34 1C, and dietary protein level ranged
from 25% to 50% (Proximate composition of the experimental
diets was determined using AOAC (2000) procedures). Each factor
has ve coded levels with this central composite design:  a, 1,
0, 1, a, respectively, with a being the star arm. The numbers of
factorial and axial points are four and four, respectively. The
number of center points was set equal to ve, thus the star arm
a 71.41421 in this case. The total number of experimental runs
was 13, and each run was replicated thrice. See Table 1 for coded
and actual combinations of temperature and dietary protein
levels.
2.3. Acclimatization of experimental sh
The acclimatization of experimental sh to temperature was
conducted in plastic buckets (1.2 m3) in a progressive fashion,
with a temperature increase or decrease of about 2 1C each day.
After acclimatized to the temperatures as set up in Table 1,
experimental sh continued to be reared for 7 days under this
regime. Temperatures were controlled using electronic thermostat rods (range 1836 1C).
2.4. Experimental management
A total of 39 plastic buckets (1.2 m3 each) were used in the
experiment, with each experimental run having three buckets or
replicates. Volume (1 m3) of tap water that had been aerated
consecutively for 3 day was added to each of the 39 plastic
buckets into which also a 300-W submersible pump was put for
the purposes of water ltration and recirculation. The temperatures were adjusted to the corresponding ones as in Table 1.

Table 1
Experimental design of temperature-dietary protein level combinations and response observations (Mean 7SD) (n 15).
Run

1
2
3
4
5
6
7
8
9
10
11
12
13

Coded

Actual

T(1C)

P (%)

0
1
0
0
1
0
0
0

0
1
a
0
1
0

27.0
22.1
27.0
27.0
22.1
27.0
27.0
27.0
34.0
20.0
31.9
27.0
31.9

37.5
28.7
25.0
37.5
46.3
37.5
50.0
37.5
37.5
37.5
46.3
37.5
28.7

a
a
1
0
1

a
0
0
0
1
0
1

SGR (%/d)

FE

Serum IGF-I (ng/mL)

Hepatic
IGF-I mRNA/b-actin mRNA

2.454 70.147
1.501 70.102
1.989 70.151
2.503 70.217
1.764 70.146
2.378 70.152
2.484 70.163
2.526 70.231
2.330 70.205
1.408 70.107
2.991 70.271
2.597 70.219
2.570 70.248

0.757 7 0.068
0.436 7 0.032
0.576 7 0.042
0.782 7 0.071
0.6057 0.055
0.8037 0.063
0.7037 0.072
0.752 7 0.061
0.629 7 0.047
0.391 7 0.026
0.683 7 0.055
0.741 7 0.058
0.772 7 0.073

28.429 74.154
17.283 72.951
24.251 73.817
28.639 74.284
17.538 73.026
30.412 74.160
22.492 73.281
28.635 73.942
23.389 73.229
15.232 71.902
23.511 72.763
29.418 74.751
29.702 74.095

3.869 7 0.927
1.5037 0.414
2.851 7 0.832
3.927 7 0.963
1.0007 0.311
3.562 7 1.027
2.0527 0.615
3.693 7 1.094
2.916 7 1.135
1.0457 0.322
2.758 7 0.681
3.722 7 0.775
3.674 7 0.892

Note: a is star arm, and 9a9 1.41421 for this experimental design.

688

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

Forty-ve sh were contained in each experimental run, with

each replicate having fteen individuals. The body weight and
body length of the experimental sh used were 27.64 (71.09 g)
and 9.43 ( 70.46 cm), respectively. Results of MANOVA showed
that there was no signicant difference between various experimental runs and between all replicates (P40.05). The feed that
with differing levels of protein was given as prescribed in Table 2.
Fish were offered experimental diets to apparent satiation three
times daily (7:00 h, 11:30 h and 16:00 h), six days a week. Fish
were considered satiated when two to three pellets feed were not
consumed within 3 min after being offered. The amount of feed
consumed by each group was subsequently calculated as summation of given diets during the experimental period. Fish in each
bucket were fortnightly group-weighed. Continuous aeration was
ensured during the entire experimental period. Fish feces were
siphoned out every day. Water (1/3) was replaced after every 3
days, with a temperature difference of 70.3 1C. Dissolved oxygen
was greater than 5 mg/L; pH was 7.470.2; ammonia-N and
nitrite were both less than 0.01 mg/L. The whole experiment
was carried out under natural photoperiod and lasted for 56 days;
from 1 November 2011 to 26 December 2011.
2.5. Measurement of response
Feeding stopped 24 h before the conclusion of experiment, and
the data measurement of all the sh within each experimental
run was done on the second day. SGR and FE were computed as
follows:
SGR%=d lnW 2 lnW 1 =t 2 t 1   100
FE W 2 W 1 =F
where W1, W2 are body weights (g) at starting (t1) and ending
time (t2) of the experiment, respectively; F is total feeding amount
(g). To avoid changes in measurement index induced by stress,
sh were killed with an overdose of tricaine methanesulfonate
(2%; MS-222) within 1 min after being captured. Blood samples
were collected from the caudal vein of ve sh in each plastic
Table 2
Ingredients and composition of experimental diets.
Ingredients

Dietary protein level (%) (g/100 g dry diet)

25.0

28.7

37.5

46.3

50.0

Fish meal
Casein
Gelatin
Corn starch
Fish oil soybean oil (1:1)
Soybean meal
cottonseed meal
Rapeseed meal
Vitamin premix1
Mineral premix2
Choline chloride
Vitamin C phosphate ester
Ca(H2PO4)2
Total

5.00
4.00
1.00
36.80
7.00
12.00
15.00
15.00
1.00
1.00
0.50
0.20
1.50
100

5.00
7.60
1.90
32.30
7.00
12.00
15.00
15.00
1.00
1.00
0.50
0.20
1.50
100

5.00
16.20
4.05
21.55
7.00
12.00
15.00
15.00
1.00
1.00
0.50
0.20
1.50
100

5.00
23.80
5.95
12.05
7.00
12.00
15.00
15.00
1.00
1.00
0.50
0.20
1.50
100

5.00
27.50
6.88
7.42
7.00
12.00
15.00
15.00
1.00
1.00
0.50
0.20
1.50
100

Proximate composition (%)

Dry matter
Crude protein
Crude lipid
Ash
Gross energy (kJ/g diet)

91.57
25.16
7.87
6.23
12.90

91.68
28.95
7.89
6.38
12.97

91.74
38.04
7.93
6.64
13.12

91.94
46.09
7.97
6.87
13.26

92.05
49.92
7.99
6.92
13.31

Note:(1) Vitamin premix (mg/kg dry diet):VA 10, VD 0.05,VE 400, VK 40, VB1 50,
VB2 200,VB3 500, VB6 50, VB7 5, VB11 15, VB12 011,VC1000, inositol 2000,
choline 5000; (2) Mineral premix (mg/kg dry diet): FeSO4  7H2O 372, CuSO4  5H2O 25,
ZnSO4  7H2O 120, MnSO4  H2O 5, MgSO4 2475, NaCl 1875, KH2PO4 1000,
Ca (H2 PO4)2 2500.

bucket. All blood samples were kept for 2 h in a refrigerator at

4 1C, were then centrifuged for 10 min at 4 1C and 3500 rpm to
obtain the serum. The supernatant was transferred and held in
refrigerator at  80 1C for further use. Another ve sh samples
were collected from each plastic bucket, and some 0.1 g liver were
obtained and immediately frozen in liquid nitrogen, then stored
at  80 1C until measurement of IGF-I mRNA levels.
2.6. Serum IGF-I level assay
Serum IGF-I was measured using the sh insulin-like growth
factor I radioimmunoassay (RIA) kit from Tianjin Nine Tripods
Medical & Bioengineering Co., Ltd. (Tianjin, China). Fish RIA kits
follow acid:ethanol extraction as described in Taylor et al. (2005).
The resultant supernatant was collected and 50 mL assay collected
for analysis, following the manufacturers protocol. The minimum
detection limit was 0.15 ng/mL, with intra- and inter-assay
coefcient of variation of 4.4% and 13.9%, respectively.
2.7. Real-time quantitative PCR (qPCR)
Tissue mRNA expression levels were determined by qPCR.
Primers for qPCR were designed with reference to the known
sequences of tilapia as given in Table 3. All primers were
synthesized by Shanghai Genecore, BioScience & Technology
Company, China. The PCR products were 100110 bp long.
We extracted total RNA from the tissue using Trizol reagent
(Dalian Takara Co. Ltd., China). RNA samples were treated with
DEPC water (Dalian Takara Co. Limited, China). The quantity of
the RNA was calculated using the absorbance at 260 nm. The
integrity and relative quantity of RNA were checked by electrophoresis. We generated cDNA from 350 ng RNA using PrimeScript
RT reagent Kit Perfect Real Time kit (Dalian Takara). PCR amplication was conducted using an ABI 7900HT Fast Real-Time PCR
System (ABI, USA) and SYBR Green PCR Master Mix (ABI),
according to the manufacturers instructions. The amplications
were performed in a 96-well plate in a 25 mL reaction volume
containing 12.5 mL of 2  SYBR Green PCR Master Mix, 0.5 mL
(each) of the forward and reverse primers (10 mM), 2 mL of
template and 9.5 mL of DEPC water. qPCR was performed as
follows: denaturizing at 95 1C for 5 min; 40 cycles for denaturizing at 95 1C for 15 s, annealing at 62 1C for 60 s. We calculated the
relative quantication of the target gene transcript IGF-I with a
chosen reference gene transcript (b-actin) using the 2-DDCT
method. This mathematical algorithm computes an expression
ratio based on real-time PCR efciency and the crossing point
deviation of the sample versus the control. We measured the PCR
efciency by constructing a standard curve using a serial dilution
of cDNA. The PCR efciency for all assay was 498%. A no
template control (NTC) and no reverse transcriptase control
(NRT) were used as control for template and genomic contamination, respectively. All samples were run in triplicate. Values of
IGF-I mRNA were then expressed relative to the lowest sample
level (control) measured within an experiment (assigned an
arbitrary value of 1).

Table 3
Primer sequences.
Target
mRNA

Sequence

NCBI Genbank
accession no.

IGF-I

F: 50 - TTGTCTGTGGAGAGCGAGGCTT-30
F: 50 - CAGCTTTGGAAGCAGCACTCGT-30
F: 50 -CCACACAGTGCCCATCTACGA-30
R: 50 -CCACGCTCTGTCAGGATCTTCA-30

XM003448059

b-actin

EU887951.1

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

689

2.8. Data processing

Experimental data were recorded in terms of mean ( 7SD). The
following form of the relationship between the responses and the
factors of interest was assumed:

2.48

where Y^ is the prediction of response (SGR, FE, serum IGF-I or

hepatic IGF-I mRNA level); b0 is regression constant; b1, b2, linear
effects of temperature and dietary protein level, respectively; b3,
interactive effect between temperature and dietary protein level;
b4, b5, quadratic effects of temperature and dietary protein level,
respectively. All these effects were estimated using the least
squares method. STATISTICA (v 8.0) Software was used for
plotting and analyzing the experimental data. Correlations
between SGR, FE, serum IGF-I and hepatic IGF-I mRNA level were
tested using Pearsons product-moment correlations of the SPSS
software (v15.0) and drawing were done using Microsoft excel.
Signicance level was set at Po0.05.

Specific growth rate (%d

Y^ b0 b1 T b2 P b3 T  P b4 T 2 b5 P 2

3.00

1.95
1.43
0.90

50.0

34.0
43.8

Die

tar

yp

Fig. 1. Response surface plot of the effect of temperature and dietary protein level
and their mutual interactions on SGR of Nile tilapia juveniles (n 15).

The experimental results of SGR are listed in Table 1, and the

least squares estimates of regression coefcients are displayed in
Table 4. Using ANOVA, the P value for the model of SGR was
0.0031, showing that the model of SGR built was of high
signicance. The P value for the lack-of-t test of the model was
0.0993, showing that the model was adequate. From Table 4, the
linear and quadratic effect of water temperature on SGR was
highly signicant (P o0.01); the linear effect of dietary protein
level on SGR was highly signicant (Po0.01), but the quadratic
effect was insignicant (P40.05); the interaction between the
two factors of interest was also not signicant (P40.05). According to the magnitude of coded coefcients, water temperature
was more crucial than dietary protein level. The actual model
equation derived was as follows:
2

SGR 8:8079 0:583T 0:0927P 0:001TP0:0096T 0:0013P

The coefcient of determination of the model R2,0.958, demonstrating that this model equation was of high goodness of t to
experimental data.
In order to visualize how SGR is simultaneously affected by
water temperature and dietary protein level, response surface
plot was drawn (Fig. 1). It could be seen clearly that under the
conditions of the experiment, SGR varied with increased water
temperature and dietary protein level in a curvilinear manner.
The SGR was greater at higher protein levels than at lower protein
Table 4
Signicance, standard error and condence interval of regression coefcients for
SGR model.
Term

Intercept
T
P
TP
T2
P2

Regression
coefcient

Standard
error

95%C.I.
Low

95% C.I.
High

P value

o
0.0001
0.0060
0.5506
0.0017
0.0710

2.492
0.485

0.056
0.045

2.358
0.380

2.625
0.591

0.173
0.040
 0.235
 0.102

0.045
0.063
0.048
0.048

0.068
 0.110
 0.348
 0.215

0.278
0.189
 0.122
0.011

37.5
27.0
)
C
e(
ein
r
31.3
23.5
u
t
lev
era
el
25.0 20.0
(%
mp
e
)
T

rot

3. Results
3.1. Effects of water temperature and dietary protein level
on SGR of Nile tilapia juveniles

30.5

Note: coefcients were given in terms of coded factors. R2 0.958, adjusted

R2 0.928.

Table 5
Signicance, standard error and condence interval of regression coefcients for
FE model.
Term

Regression
coefcient

Standard
error

95%C.I.
Low

95%C.I.
High

P value

Intercept
0.767
T
0.094
P
0.032
TP
 0.065
T2
 0.116

0.016
0.013
0.013
0.018
0.014

0.728
0.063
0.002
 0.108
 0.149

0.806
0.124
0.063
 0.021
 0.083

P2

0.014

 0.084

 0.019

0.0002
0.0404
0.0096
o
0.0001
0.0076

 0.051

Note: coefcients were given in terms of coded factors. R2 0.955, adjusted

R2 0.923.

levels regardless of water temperature differences. For example,

SGR was notably higher when protein level was 45% than when
protein level was 25%, with water temperature held at 27 1C.

3.2. Effects of water temperature and dietary protein level on FE of

Nile tilapia juveniles
The observed data of FE are provided in Table 1, and the model
coefcients estimated using least squares approach are presented
in Table 5. It was found via ANOVA that the model of FE was
highly signicant (P 0.0001); the P value for the lack-of-t test
was 0.1231, showing that the model of FE was of great adequacy;
the linear and quadratic effects of water temperature on FE were
all highly signicant (P o0.01); the linear effect of dietary protein
level on FE was signicant (Po0.05), and the quadratic effect was
highly signicant (Po0.01); the interaction between the two
factors was also highly signicant (P o0.01). Water temperature
was more important than dietary protein level in inuencing FE.
The actual model equation of FE attained was as follows:
FE 5:7582 0:3303T 0:0929P0:0015TP0:0047T 2 0:0007P2

with the coefcient of determination as high as R2 0.955.

690

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

From Fig. 2, it could be seen that FE varied curvilinear with

both water temperature and dietary protein level. For instance,
with water temperature ranging between 20 1C and 29 1C and
protein level kept at 37.5%, FE increased as water temperature
increases; when water temperature was higher than 29 1C, FE
began to decrease. With water temperature held at 27 1C and
protein level ranged between 25% and 40%, FE gradually increased
as dietary protein level increases; as dietary protein levels were
higher than 40%, FE began to decrease. When water temperature
and protein level lay at the combination of 28.9 1C/38.2%, a larger
FE, 0.786, was reached.
3.3. Effects of water temperature and dietary protein level on serum
IGF-I level in Nile tilapia juveniles
Estimates of regression coefcients and their signicance
(Table 6) were obtained through statistical analysis for the
experimental results of serum IGF-I level, as shown in Table 1.
Outputs of ANOVA showed that the model equation of serum
IGF-I level was highly signicant (Po0.01) and adequate
(P0.1042 for the lack-of-t test); the linear and quadratic effects
of water temperature on serum IGF-I level were highly signicant
(Po0.01); the linear effect of dietary protein level on serum

IGF-I was signicant (Po 0.05), and the quadratic effect proved
highly signicant (Po0.01); the interaction between the two
factors was also signicant (Po0.05); water temperature was
more critical than dietary protein level in affecting serum IGF-I.
The actual model equation secured was as follows:
Serum IGF-I 213:5159 12:5650T 3:4680P
0:0368TP0:1931T 2 0:0346P2
the coefcient of determination arrived at was R2 0.967
It could be seen from Fig. 3 that serum IGF-I level varied with
both water temperature and dietary protein level in a curvilinear
fashion. For example, with water temperature ranged between
20 1C and 29 1C and dietary protein level kept at 37.5%, serum
IGF-I level increased as water temperature increases; but when
water temperature was higher than 29 1C, serum IGF-I level began
to decrease. At water temperature of 27 1C and dietary protein
level between 25% and 36%, serum IGF-I level gradually increased
as dietary protein level was increasing; while dietary protein level
higher than 36%, resulted to a gradual decrease in serum IGF-I.
When water temperature and dietary protein level lay at the
combination, 29.2 1C/34.6%, a larger serum IGF-I level, 30.122 ng/mL,
was observed.
3.4. Effects of water temperature and dietary protein level on the
hepatic IGF-I mRNA level of Nile tilapia juveniles

79.00

The observed results of hepatic IGF-I mRNA levels are provided

in Table 1, and the model coefcients estimated using least
squares approach are presented in Table 7. It was found via
ANOVA that the model of hepatic IGF-I mRNA levels was highly
signicant (P 0.0001); the P value for the lack-of-t test was
0.1334, showing that the model of hepatic IGF-I mRNA levels was
of great adequacy. The linear effect of water temperature on IGF-I
mRNA levels was highly signicant (Po0.01), but the quadratic
effect was insignicant (P 40.05); the linear and quadratic effect
of dietary protein level on IGF-I mRNA levels was highly signicant (Po0.01); and the interaction between the two factors of
interest was also highly signicant (Po0.01). Water temperature
was more crucial than dietary protein level. The following actual

45.50
28.75
12.00

50.0

34.0

Fig. 2. Response surface plot of the effect of temperature and dietary protein level
and their mutual interactions on FE of Nile tilapia juveniles (n 15).

Table 6
Signicance, standard error and condence interval of regression coefcients for
serum IGF-I model.
Term

Intercept
T
P
TP
T2
P2

Regression
coefcient
29.107
3.741
 1.053
 1.612
 4.731
 2.701

Standard
error
0.559
0.442
0.442
0.625
0.474
0.474

95%C.I.
Low
27.784
2.695
 2.099
 3.091
 5.853
 3.822

95%C.I.
High
30.430
4.787
 0.007
 0.132
 3.610
 1.579

31.00

el (ng/ml)

30.5
Die 43.8
tar
)
yp
37.5
27.0
C
rot
e(
ein
r
u
31.3
t
23.5
lev
era
el
(%
mp
25.0 20.0
e
)
T

Serum IGF-I lev

Feed efficiency

62.25

25.00
19.00
13.00
7.00

P value

o
0.0001
0.0488
0.0367
o
0.0001
0.0007

Note: coefcients were given in terms of coded factors. R2 0.967, adjusted

R2 0.943.

50.0
Die 43.8
tar
y p 37.5
rot
ein
lev 31.3
el
(%
)

34.0
30.5

27.0
23.5
25.0 20.0

re
atu

(C

er

mp

Te

Fig. 3. Response surface plot of the effect of temperature and dietary protein level
and their mutual interactions on serum IGF-I level of Nile tilapia juveniles (n 15).

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

Table 7
Signicance, standard error and condence interval of regression coefcients for
hepatic IGF-I mRNA model.
Term

Regression
coefcient

Standard
error

95%C.I.
Low

95%C.I.
High

P value

Intercept
T
P
TP
T2
P2

4.755
0.822
 0.319
 0.103
 0.883
 0.647

0.093
0.073
0.073
0.104
0.079
0.079

4.535
0.648
 0.492
 0.349
 1.069
 0.833

4.974
0.995
 0.145
0.142
 0.697
 0.461

o 0.0001
o 0.0001
0.0034
0.3529
o 0.0001

Note: coefcients were given in terms of coded factors. R2 0.978, adjusted

R2 0.963.

4.00

691

3.6. Relationship of serum IGF-I level and hepatic IGF-I mRNA level
to SGR and FE
Serum IGF-I level was found to be positively correlated with
SGR (Fig. 5), with correlation coefcient r 0.768 (Po0.01). The
regression of serum IGF-I level towards SGR was serum IGFI 8.6416 SGR 4.9267
Hepatic IGF-I mRNA level was found to be positively correlated
with SGR (Fig. 6), with correlation coefcient r 0.751 (Po0.01).
The regression of hepatic IGF-I mRNA level towards SGR was
hepatic IGF-I mRNA level 1.7195SGR  1.1146.
Serum IGF-I level was also found to be positively correlated
with FE (Fig. 7), with correlation coefcient r 0.911 (Po0.01).
The regression of serum IGF-I level towards FE was serum IGF-I
36.189 FE 0.509.
Hepatic IGF-I mRNA level was also found to be positively
correlated with FE (Fig. 8), with correlation coefcient r 0.835
(P o0.01). The regression of hepatic IGF-I mRNA level towards FE
was hepatic IGF-I mRNA level 6.8477 FE  1.7326.

4. Discussion
1.60

4.1. Effects of water temperature on SGR and FE in Nile tilapia

juveniles

0.40

Water temperature plays a crucial role in food digestibility in

sh. Higher temperature contributes to increased food intake and
feed utilization, and in turn to intestinal digestion and absorption

-0.80

50.0

35

34.0

30.5
Die 43.8
tar
)
yp
37.5
27.0
(C
rot
e
r
ein
tu
23.5
lev 31.3
era
el
mp
25.0 20.0
e
(%
T
)
Fig. 4. Response surface plot of the effect of temperature and dietary protein level
and their mutual interactions on hepatic IGF-I mRNA level of Nile tilapia juveniles
(n 15).

Serum IGF-I levels (ng/mL)

Hepatic IGF-I mRNA

/-actin mRNA

2.80

28
21
14
Serum IGF-I = 8.6416SGR + 4.9267
r = 0.768

model equation was obtained:

Hepatic IGFI mRNA levels 38:6765 2:1999T 0:6489P

0
1.2

1.5

0:0024T  P0:0367T 2 0:0083P 2

3.5. Simultaneous optimization of responses

To obtain the optimal level combination of the two factors, the
model equations of the SGR and FE were simultaneously optimized, following the procedure of Montgomery (2005). It was
found that the optimum level combination of water temperature
and dietary protein level was 29.9 1C/40.3%, at which the greatest
SGR and FE were simultaneously reached, i.e., 2.748%/d and 0.775,
respectively, with a desirability value of 0.888.

3.3

Fig. 5. Relationship between serum IGF-I level and SGR of Nile tilapia juveniles.
Data from three replicates are mean values (n 15).

5
Hepatic IGF-I mRNA
/-actin mRNA

with the coefcient of determination: R2 0.978.

From Fig. 4 it is observed that IGF-I mRNA levels of juveniles
rst increased with water temperature at 37.5% protein level, and
then decreased when water temperature exceeded 29 1C. IGF-I
mRNA levels of juveniles varied curvilinear as dietary protein
level changed linearly. Hepatic IGF-I mRNA levels of tilapia
juveniles was obviously subdued by low temperature, low and
high dietary protein level.

1.8
2.1
2.4
2.7
Specific growth rate (%/d)

4
3
2
IGF-I mRNA = 1.7195 SGR - 1.1146
r = 0.751

1
0
1.2

1.5

1.8

2.1

2.4

2.7

3.3

Specific growth rate (%/d)

Fig. 6. Relationship between hepatic IGF-I mRNA level and SGR of Nile tilapia
juveniles. Data from three replicates are mean values (n 15).

692

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

Serum IGF-I level (ng/mL)

35
28
21
14
Serum IGF-I = 36.189FE + 0.509
r = 0.911

7
0
0.3

0.4

0.5

0.6

0.7

0.8

0.9

Feed efficiency
Fig. 7. Relationship between serum IGF-I level and FE of Nile tilapia juveniles.
Data from three replicates are mean values (n 15).

Hepatic IGF-I mRNA

/-actin mRNA

IGF - I mRNA = 6.8477 FE - 1.7326

r = 0.835

4
3
2
1
0
0.3

0.4

0.5

0.6
0.7
Feed efficiency

0.8

0.9

Fig. 8. Relationship between hepatic IGF-I mRNA level and FE of Nile tilapia
juveniles. Data from three replicates are mean values (n 15).

(Reid et al., 1995; Musuka et al., 2009). In the present study, the
effect of water temperature on SGR and FE in tilapia juveniles
proved very signicant. For example, when dietary protein level
maintained at 37.5%, the SGR and FE were respectively 2.378
2.597%/d and 0.7410.803 at water temperature 27 1C, notably
higher than 1.408%/d and 0.391 at water temperature 20 1C. This
is analogous with the nding of Musuka et al. (2009). Response
surface analysis showed that when dietary protein level was
37.5%, the greatest SGR and FE reached were 2.706%/d and
0.779 at 30.2 1C respectively. This water temperature falls within
the suitable temperature range, 2931 1C, reported by Popma and
Lovshin (1995) for tilapia growth. It can be seen that less energy
was used for maintaining shs internal requirements at optimal
water temperature, growth was thus effectively promoted. High
water temperature, above 30 1C for example, may pose physiological stress in sh, and the energy produced from the resulting
metabolism is primarily used to sustain homeostasis. At the same
time, high temperature could be attributed to the high rate of
gastric evacuation leading to retard growth and poor feed utilization (Elliot, 1972). Azaza et al. (2008) further reported that
growth rate of Nile tilapia was better at 30 1C than at 34 1C.
4.2. Effects of dietary protein level on SGR and FE
in Nile tilapia juveniles
As dietary protein level increased, tilapia juveniles were found
to grow faster in the study. When water temperature held at
27 1C, SGR value was 2.565%/d at dietary protein level of 45%,
noticeably higher than 1.989%/d at 25%; high dietary protein level

of 45% did not restrain growth in Nile tilapia. This is different

from the result of Abdel-Tawwab et al. (2010), who found that at
water temperature of 27 1C, SGR value was 1.107%/d at dietary
protein level of 45%, slightly lower than 1.143%/d at 35%. The
impact of dietary proteins on the growth of tilapia may be closely
linked to other factors such as rearing density, strain, number of
generation of selection and environment (Al Hafedh, 1999). In this
experiment, the Nile tilapia juveniles used were of the 16th
generation of GIFT strain, and furthermore four rounds of mass
selection have been carried out on them in Yixing experimental
Base, China. When dietary protein level reached 50%, growth can
be inhibited in Nile tilapia. Juancey (1982) also found that, the
Mozambique tilapia Oreochromis mossambicus fed diets of higher
protein level (56%) showed poor growth rate than those fed with
lower protein level of 48%. However, when dietary protein level
was 25%, most of the proteins may have being utilized to
maintain the organisms energy requirements, and the quantity
of essential amino acids to be used for sh growth dwindled.
Alternatively, the higher level of dietary carbohydrates may play
an inhibitive role in tilapia growth (Perez-Sanchez et al., 1995).
Dietary protein level also had signicant impact on FE of
tilapia juveniles. For instance, at a water temperature of 27 1C,
FE increased as dietary protein level varied from 25% to 40% and is
akin to the result reported by Al Hafedh (1999). When dietary
protein level went beyond 40%, FE decreased with increased
dietary protein level. This is similar to the nding reported by
Abdel-Tawwab et al. (2010). Maybe the dietary protein level went
out of the suitable range required; and excessive amino acids
were unable to be normally absorbed. Therefore they are either
transformed into fats accumulating in tissue such as muscle, or
excreted as urine nitrogen and hence caused a waste of proteins
(Alvarez-Gonzalez et al., 2001).
4.3. Change in IGF-I at varying levels of water temperature and
its relationship with SGR and FE
The IGF-I/GH axis of sh species, like that of vertebrates, plays
an important role in sex differentiation, growth and reproduction
(Cameron et al., 2007), whose activity can be affected by both
biological functions of sh organs and external factors (Duan,
1997). IGF-I is mainly produced in liver, which is also the main
source of IGF-I hormone (Debnath, 2010). This study found, water
temperature to be closely related to tilapia serum IGF-I and
hepatic IGF-I mRNA level, and higher water temperature helped
increase serum IGF-I and hepatic IGF-I mRNA level. Vera Cruz
et al. (2006) also found that hepatic IGF-I mRNA level was
signicantly correlated with temperature conditions in Nile
tilapia. Similar results were found in other sh species such as
coho salmon, Oncorhynchus kisutch (Duan et al., 1995), channel
catsh (Silverstein et al., 2000), gilthead sea bream, Sparus aurata
(Mingarro et al., 2002), rainbow trout, Oncorhynchus mykiss
(Gabillard et al., 2003b; Imsland et al., 2007), and halibut
Hippoglossus hippoglossus (Imsland et al., 2008). The serum IGF-I
level was found to be closely correlated with SGR (Fig. 5), with the
Pearson correlation r 0.768 (Po0.01). Analogous ndings of this
relationship have also been reported in barramundi, Lates calcarifer, by Matthews et al. (1997); in coho salmon by Pierce et al.
(2001); in Mozambique tilapia, by Uchida et al. (2003) and in
turbot, by Imsland et al. (2007). At the same time, hepatic IGF-I
mRNA level was signicantly related to SGR (Fig. 6), and this
result was similar to Vera Cruz et al. (2006). However, when the
water temperature was 34 1C, high temperature suppressed
serum IGF-I and hepatic IGF-I mRNA level from our study.
Improved SGR and FE at 30 1C inferred that, this temperature
perhaps was approaching the thermal optima for Nile tilapia;
with 34 1C possibly exceeding it, whereby high temperatures

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

decrease biochemical activity or production rates, that may lead

to reduction in circulating IGF-I. Gabillard et al. (2005) pointed
out that optimal temperature could hasten growth and release of
GH in rainbow trout, and serum IGF-I was positively correlated
with GH, which in turn could increase hepatic IGF-I mRNA level
and serum IGF-I.
Mingarro et al. (2002) also found that serum IGF-I level
increased in temperate sh during suitable season for their
growth. IGF-I, as a cytokinin, can catalyze the ornithine dehydrogenase activity, expedite the intracellular biosynthesis of DNA,
RNA and proteins, and nally cause the proliferation and differentiation of cells, thereby contributing to growth (Tsai et al.,
1994). The FE was for the rst time, found to be positively
correlated with the serum IGF-I and hepatic IGF-I mRNA level of
Nile tilapia juveniles, the Pearson correlation was r 0.911 and
r 0.835 receptively (Po0.01). This is similar to the nding
reported by Pierce et al. (2001) and Luckenbach et al. (2007).
However, this correlation was not found in the study of Imsland
et al. (2007).
4.4. Effect of dietary protein level on serum IGF-I
and hepatic IGF-I mRNA level
As an important hormone, IGF-I has many functional tendencies, such as regulating cellular metabolism, promoting cellular
growth, division and differentiation, enhancing the development
of oosperm, mediating growth and osmotic pressure adaptation
(Mendez et al., 2000; Gabillard et al., 2003a). In this study, dietary
protein level was found to have signicant impact on the serum
IGF-I and hepatic IGF-I mRNA level in tilapia juveniles. Dyer et al.
(2004) found that plasma IGF-I level was positively correlated
with growth rate and dietary protein level in barramundi (Lates
calcarifer), with r2 0.65 and 0.59, respectively. Perez-Sanchez
et al. (1995) reported that plasma IGF-I level increased with
increasing dietary protein level in gilthead sea bream (Sparus
aurata). IGF-I might provide a useful tool for monitoring sh
growth and evaluating nutritional status (Ma et al., 1992;
Wilkinson et al., 2006). Jiang et al. (2010) also found that dietary
protein level could noticeably affect the hepatic IGF-I mRNA level
in mud carp (Cirrhinus molitorella), and SGR was signicantly
correlated with the serum IGF-I level. Different sources of protein
can signicantly affect serum IGF-I and hepatic IGF-I mRNA level
in sh. Gomez-Requeni et al. (2004) reported that total or partial
replacement of sh meal by a mixture of plant protein sources
could lead to decreased growth, reduced plasma IGF-I and hepatic
IGF-I mRNA level in juvenile gilthead sea bream.

693

the dietary proteins. Thus, the demand and utilization of dietary

protein in tilapia diet changed at different water temperature.
These changes may alter the GH/IGF-I axis, and in turn impact on
growth and feed utilization.
In this study, the quadratic effect of water temperature on SGR
(Table 4), FE (Table 5) and serum IGF-I level (Table 6) and the
quadratic effect of dietary protein level on FE, serum IGF-I level
and hepatic IGF-I mRNA level (Table 7) were found to be
signicant for the rst time. This may be of great value in tilapia
production, because when water temperature and dietary protein
level deviate from the optimal factor combination, the
response(s) of interest would be declining rapidly. Therefore, if
the rearing of tilapia juveniles could be arranged according to the
optimal factor combination derived in this study, the production
efciency would be sharpened.
4.6. Model building and response optimization
The factorial design was used mainly in previous studies that
dealt with the relationship of SGR and FE with water temperature
and/or dietary protein level, whereas in our study the central
composite design was adopted. Using the central composite design,
the effect of the factors of interest can be examined in concert
rather than singly; the predictive model equations can be established, and the optimal factor combination also can be determined
through simultaneous optimization of response models. In this
study, the model equations of SGR, FE, serum IGF-I level and
hepatic IGF-I mRNA level using least squares method were
obtained, with the coefcients of determination R2 0.958, 0.955,
0.967, 0.978, respectively. These models can be used for practical
projection. Using the procedure of Montgomery (2005), the model
equations of SGR and FE were simultaneously optimized, and the
optimal factor combination of water temperature and dietary
protein level, i.e., 29.9 1C/40.3%, was obtained, at which the largest
SGR and FE were 2.748%/d and 0.775, respectively, with the
desirability function value of 0.888. It should be seen, however,
that in practical production, due to the increasing sh meal prices,
a higher level of dietary protein may bring about an increasing cost
of culture. Increasing dietary lipid level or determining optimal
ratio of protein to carbohydrate may help improve the utilization
of dietary nitrogen and diminish the waste of protein. Other factors
such as salinity, pH, dissolved oxygen and photoperiod may play a
combined role in the utilization of dietary proteins in tilapia. This
should be further examined.

Acknowledgments
4.5. Interaction and quadratic effects of water temperature
and dietary protein level on SGR, FE and serum IGF-I and
hepatic IGF-I mRNA level
The growth of sh is an intricate process which is affected by
both biotic and abiotic factors such as nutrition, temperature,
rearing density and salinity. The interaction among these factors
may occur. In our study, the interaction between water temperature and dietary protein level on FE, serum IGF-I and hepatic IGF-I
mRNA level was signicant in Nile tilapia, but that on SGR was
insignicant, such has not being found or reported by others.
When water temperature is relatively lower, higher demand for
dietary proteins or amino acids may be required by tilapia
juveniles to maintain their energy requirement. At higher water
temperature, say 27 1C, which may be more suitable for the
growth of tilapia juveniles, more dietary proteins may be used
for growth. Whereas in high water temperature environment,
physiological stress usually occurs within sh and the activity of
proteinase is prohibited (Han et al., 2011), thereby underutilizing

The study was nanced by the following grants: Special Fund for
Agro-scientic Research in the Public Interest (200903046-02);
Special Fund for Modern Agricultural Industry Technology System
Construction-Tilapia Industry Technology System (CARS-49); Postgraduate Scientic Research Innovation Program of Jiangsu Ordinary
Higher Colleges and Universities (CXLX11-0708); Special Fund for
Guangdong Marine Fisheries Science & Technology Extension Program (A201009C02, A2010002-010b); Guangdong Science & Technology Program (2010B090500032); National Science & Technology
Program (2012BAD26B00). The authors are also grateful to the
director, Mr. Zhu Zhixiang, of the Yixing Experimental Base of
Chinese Academy of Fishery Sciences, for providing us with biological material, site and other technical assistance.
References
Abdel-Tawwab, M., Ahmad, M.H., Khattab, Y.A.E., Shalaby, A.M.E., 2010. Effect of
dietary protein level, initial body weight, and their interaction on the growth,

694

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

feed utilization, and physiological alterations of Nile tilapia, Oreochromis

niloticus (L.). Aquaculture 298, 267274.
Al Hafedh, Y.S., 1999. Effects of dietary protein on growth and body composition of
Nile tilapia. Aquaculture Res. 30, 385393.
Alvarez-Gonzalez, C.A., Civera-Cerecedo, R., Ortiz-Galindo, J.L., Dumas, S., MorenoLegorreta, M., Grayeb-Del Alamo, T., 2001. Effect of dietary protein level on
growth and body composition of juvenile spotted sand bass, Paralabrax
maculatofasciatus, fed practical diets. Aquaculture 194, 151159.
AOAC, 2000. Ofcial Methods of Analysis 17th Ed. Association of the Ofcial
Analytical Chemists, Washington, DC, USA.
Azaza, M.S., Dhraief, M.N., Kraiem, M.M., 2008. Effects of water temperature on
growth and sex ratio of juvenile Nile Tilapia Oreochromis niloticus (Linneaus)
reared in geothermal waters in southern Tunisia. J. Therm. Biol. 192, 132145.
os, N., Planas, J.V., Gutierrez, J., Navarro, I., 1999. Regulation of plasma insulinBan
like growth factor-I levels in brown trout (Salmo trutta). Comp. Biochem.
Physiol. C 124, 3340.
Beckman, B.R., Larsen, D.A., Moriyama, S., Leepawlak, B., Dickhoff, W.W., 1998.
Insulin-like growth factor-I and environmental modulation of growth during
smoltication of spring Chinook salmon (Oncorhynchus tshawytscha). Gen.
Comp. Endocrinol. 109, 325335.
Beckman, B.R., Shimizu, M., Gadberry, B.A., Parkins, P.J., Cooper, K.A., 2004. The
effect of temperature change on the relations among plasma IGF-I, 41-kDa
IGFBP, and growth rate in postsmolt coho salmon. Aquaculture 241, 601619.
Cameron, C., Moccia, R., Azevedo, P.A., Leatherland, J.F., 2007. Effect of diet and
ration on the relationship between plasma GH and IGF-1 concentrations in
Arctic charr, Salvelinus alpinus (L.). AquacultureRes. 38, 877886.
Carmona-Osalde, C., Olvera-Novoa, M.A., Rodrguez-Serna, M., 2005. Effect of the
proteinlipids ratio on growth and maturation of the craysh Procambarus
(Austrocambarus) llamasi. Aquaculture 250, 692699.
Cruz, E.M.V., Brown, C.L., 2009. Inuence of the photoperiod on growth rate and
insulin-like growth factor-I gene expression in Nile tilapia Oreochromis
niloticus. J. Fish Biol. 75, 130141.
Davis, K.B., McEntire, M., 2006. Effect of photoperiod on feeding, intraperitoneal
fat, and insulin-like growth factor-I in sunshine bass. J. World Aquacult. Soc.
37, 431436.
Davis, K.B., McEntire, M., 2011. Inuence of reproductive status, sex hormones and
temperature on plasma IGF-I concentrations in sunshine bass (Morone chrysops
 Morone saxatilis). Comp. Biochem. Physiol. A 158, 1316.
Debnath, S., 2010. A review on the physiology of Insulin like Growth Factor-I
(IGF-I) peptide in bony shes and its phylogenetic correlation in 30 different
taxa of 14 families of teleosts. Adv. Environ. Biol. 5, 3152.
Duan, C., 1997. The insulin-like growth factor system and its biological actions in
sh. Am. Zool. 37, 491503.
Duan, C., Plisetskaya, E.M., Dickhoff, W.W., 1995. Expression of insulin-like growth
factor I in normally and abnormally developing coho salmon (Oncorhynchus
kisutch). Endocrinology 136, 446452.
Dyer, A.R., Barlow, C.G., Bransden, M.P., Carter, C.G., Glencross, B.D., Richardson, N.,
Thomas, P.M., Williams, K.C., Carragher, J.F., 2004. Correlation of plasma IGF-I
concentrations and growth rate in aquacultured nsh a tool for assessing the
potential of new diets. Aquaculture 236, 583592.
El-Sayed, A.M., Kawanna., M., 2008. Effects of dietary protein and energy levels on
spawning performance of Nile tilapia (Oreochromis niloticus) broodstock in a
recycling system. Aquaculture 258, 179184.
Elliot, J.M, 1972. Rates of gastric evacuation in brown trout, Salmo trutta(L.).
Freshwater Biol. 2, 118.
Enes, P., Sanchez-Gurmachesc, J., Navarroc, I., Gutierrezc, J., Oliva-Teles, A., 2010.
Role of insulin and IGF-I on the regulation of glucose metabolism in European
sea bass (Dicentrarchus labrax) fed with different dietary carbohydrate levels.
Comp. Biochem. Physiol. A 157, 346353.
Gabillard, J.C., Weil, C., Rescan, P.Y., Navarro, I., Gutierrez, J., Le Bail, P.Y., 2003a.
Environmental temperature increases plasma GH levels independently
of nutritional status in rainbow trout (Oncorhynchus mykiss). Gen. Comp.
Endocrinol. 133, 1726.
Gabillard, J.C., Weil, C., Rescan, P.Y., Navarro, I., Gutierrez, J., Le Bail, P.Y., 2003b.
Effects of environmental temperature on IGF1, IGF2, and IGF type I receptor
expression in rainbow trout (Oncorhynchus myskiss). Gen. Comp. Endocrinol.
133, 233242.
Gabillard, J.C., Weil, C., Rescan, P.Y., Navarro, I., Gutierrez, J., Le Bail, P.Y., 2005.
Does the GH/IGF system mediate the effect of water temperature on sh
growth? A review. Cybium 29, 107117.
Garcia, J.A., Villarroel, M., 2009. Effect of feed type and feeding frequency on
macrophage functions in tilapia (Oreochromis niloticus L.). Fish Shellsh
Immunol. 27, 325329.
Gunasekera, R.M., Lam, T.J., 1997. Inuence of dietary protein level on ovarian
recrudescence in Nile tilapia, Oreochromis niloticus (L.). Aquaculture 149,
5769.
Gomez-Requeni, P., Mingarro, M., Calduch-Giner, J., Medale, F., Martin, S., Houlihan, D., Kaushik, S., Perez-Sanchez, J., 2004. Protein growth performance,
amino acid utilisation and somatotropic axis responsiveness to sh meal
replacement by plant protein sources in gilthead sea bream (Sparus aurata).
Aquaculture 220, 493510.
Han, Q., Liu, L.G., Zhang, J.P., Zhao, D.H., Pan, Z., Wu, R.N., 2011. The effect of
temperature and PH on activities of digestive enzymes in catsh (Silurus asotus
linnaeus) in Dongting lake area. Acta Hydr. Sin. 34, 2229.

Hidalgo, F., Alliot, E., 1988. Inuence of water temperature on protein requirement
and protein utilization in juvenile Sea bass, Dicentrarchus labrax. Aquaculture
72, 115129.
Hossain, M.A., Almatar, S.M., James, C.M, 2010. Optimum dietary protein level for
juvenile silver pomfret, Pampus argenteus (Euphrasen). J. World Aquacult. Soc.
41, 710720.

Imsland, A.K., Bjornsson,

B.T., Gunnarsson, S., Foss, A., Stefansson, S.O, 2007.
Temperature and salinity effects on plasma insulin-like growth factor-I
concentrations and growth in juvenile turbot (Scophthalmus maximus). Aquaculture 271, 546552.
Imsland, A.K., Foss, A., Rothc, B., Stefanssonc, S.O., Vikingstadb, E., Pedersene, S.,
Sandvike, T., Norberg, B., 2008. Plasma insulin-like growth factor-I concentrations and growth in juvenile halibut (Hippoglossus hippoglossus): effects of
photoperiods and feeding regimes. Comp. Biochem. Physiol. A 151, 6670.
Jiang, J., Zhang, D., Qiu, L., Lin, H., Jiang, S., 2010. Research on assessing effects of
diets of mud carp (Cirrhinus molitorella) using IGF-I mRNA expression level.
South China Fish. Sci. 6, 6672.
Likongwe, J.S., Stecko, T.D., Stauffer, J.R., Carline, R.F., 1996. Combined effects of
water temperature and salinity on growth and feed utilization of juvenile Nile
tilapia Oreochromis niloticus (Linneaus). Aquaculture 146, 3746.
Luckenbach, J.A., Murashige, R., Daniels, H.V., Godwin, J., Borski, R.J., 2007.
Temperature affects insulin-like growth factor I and growth of juvenile
southern ounder, Paralichthys lethostigma. Comp. Biochem. Physiol. A 146,
95104.
Ma, L., Burton, K.A., Saunders, J.C., Dauncey, M.J., 1992. Thermal and nutritional
inuences on tissue levels of insulin-like growth factor-I mRNA and peptide.
J. Therm. Biol. 17, 8995.
Matthews, S.J., Kinhult, A.K., Hoeben, P., Sara, V.R., Anderson, T.A., 1997. Nutritional
regulation of insulin-like growth factor-I mRNA expression in barramundi, Lates
calcarifer. J. Mol. Endocrinol. 18, 273276.
Mendez, E., Castillo, J., Smith, A., Capilla, E., Gabillard, J.C., Planas, J.V., Navarro, I.,
Gutierrez, J., 2000. IGF I and IGF-II throughout sh development. Comp.
Biochem. Physiol. A 126, 64.
Montgomery, D.C., 2005. Design and Analysis of Experiments, 6th ed. John Wiley &
Sons, Inc., New York, USA, pp. 405444.
Mingarro, M., Vega-Rubin de Celis, S., Astola, A., Pendon, C., Valdivia, M.M.,
Perez-Sanchez, J., 2002. Endocrine mediators of seasonal growth in gilthead
sea bream (Sparus aurata): the growth hormone and somatolactin paradigm.
Gen. Comp. Endocrinol. 128, 102111.
Musuka, C.G., Likongwe, J.S., Kangombe, J., Jere, W.W.L., Mtethiwa, A.H., 2009.
The effect of dietary protein and water temperatures on performance of T.
rendalli Juveniles reared in indoor tanks. Pak. J. Nutr. 8, 15261531.
Omar, E., Al-Sagheer, F.M., Nour, A.M., Abou Akkada, A.R., 1997. Effect of protein
level and stocking density on growth performance, feed utilization and
resistance of Nile tilapia (Oreochromis niloticus) to infection against aeromonas
septicemia (Aeromonas hydrophila) Cahiers. Opt. Med. 22, 6777.
Peres, H., Oliva-Teles, A., 1999. Inuence of temperature on protein utilization in
juvenile European seabass (Dicentrarchus labrax). Aquaculture 170, 337348.
Perez-Velazquez, M., Gonzalez-Felix, M.L., Jaimes-Bustamente., F., Martnez-Cordova, L.R., Trujillo-Villalba, D.A., Davis, D.A., 2007. Investigation of the effects
of salinity and dietary protein level on growth and survival of Pacic White
Shrimp, Litopenaeus vannamei. J. World Aquaculture Soc. 38, 475485.
Petersona, B.C., Bilodeau-Bourgeois, A.L., Smalla, B.C., 2009. Response of the
somatotropic axis to alterations in feed intake of channel catsh (Ictaluruspunctatus). Comp. Biochem. Physiol. A 153, 457463.
Pierce, A.L., Beckman, B.R., Schearer, K.D., Larsen, D.A., Dickhoff, W.W., 2001.
Effects of ration on somatotropic hormones and growth in coho salmon. Comp.
Biochem. Physiol. B 128, 255264.
Pirozzi, I., Booth, M.A., Allan, G.L., 2010. The interactive effects of dietary protein
and energy on feed intake, growth and protein utilization of juvenile mulloway (Argyrosomus japonicus). Aquaculture Nutr. 16, 6171.
os, N., Capilla, E., Navarro, I., Gutierrez, J., 2000. Insulin
Planas, J.V., Mendez, E., Ban
and IGF-1 receptors in trout adipose tissue are physiologically regulated by
circulating hormone levels. J. Exp. Biol. 203, 11531159.
Popma, T.J., Lovshin, L.L., 1995. Worldwide Prospects for Commercial Production of
Tilapia, International Center for Aquaculture and Aquatic Environments
Department of Fisheries and Allied Aquacultures Auburn University, Alabama,
p. 9.
Perez-Sanchez, J., Mart-Palanca, H., Kaushik, S.J., 1995. Ration size and protein
intake affect circulating growth hormone concentration, hepatic growth
hormone binding and plasma insulin-like growth factor-I immunoreactivity
in a marine teleost, the gilthead sea bream (Sparus aurata). J. Nutr. 125,
546552.
Reid, S.D., Dockray, J.J., Linton, T.K., McDonald, D.G., Wood, C.M., 1995. Effects of a
summer temperature regime representative of a global warming scenario on
growth and protein synthesis in hardwater- and softwater-acclimated juvenile
rainbow trout (Oncorhynchus mykiss). J. Therm. Biol 20, 231244.
Riley Jr., L.G., Walker, A.P., Dorough, C.P., Schwandt, S.E., Grau, E.G., 2009. Glucose
regulates ghrelin, neuropeptide Y, and the GH/IGF-I axis in the tilapia,
Oreochromis mossambicus. Comp. Biochem. Physiol. A 154, 541546.
Silverstein, J.T., Wolters, W.R., Shimizu, M., Dickhoff, W.W., 2000. Bovine growth
hormone treatment of channel catsh: strain and temperature effects on
growth, plasma IGF-I levels, feed intake and efciency and body composition.
Aquaculture 190, 7788.

J. Qiang et al. / Journal of Thermal Biology 37 (2012) 686695

Singh, R.K., Balange, A.K., Ghughuskar, M.M., 2006. Protein sparing effect of
carbohydrates in the diet of Cirrhinus mrigala (Hamilton,1822) fry. Aquaculture 258, 680684.
Singh, R.K., Chavan, S.L., Desai, A.S., Khandagale, P.A., 2008. Inuence of dietary
protein levels and water temperature on growth, body composition and
nutrient utilization of Cirrhinus mrigala (Hamilton, 1822) fry. J. Therm. Biol.
33, 2026.
Singh, R.K., Desai, A.S., Chavan, S.L., Khandagale., P.A., 2009. Effect of water
temperature on dietary protein requirement, growth and body composition
of Asian catsh, Clarias batrachus fry. J. Therm. Biol. 34, 813.
Streit Jr, D.P., Tesser, M.B., Burkert, D., Sanchez, C.C., Sampaio, L.A, 2010. Survival
and growth of juvenile marine pejerrey, Odontesthes argentinensis, reared at
different temperatures. J. World Aquacult. Soc. 41, 931935.
Taylor, J.F., Migaud, H., Porter, M.J.R., Bromage, N.R., 2005. Photoperiod inuences
growth rate and plasma insulin-like growth factor-I levels in juvenile rainbow
trout, Oncorhynchus mykiss. Gen. Comp. Endocrinol. 142, 169185.
Taylor, J.F., Porter, M.J.R., Bromage, N.R., Migaud, H., 2008. Relationships between
environmental changes, maturity, growth rate and plasma insulin-like growth
factor-(IGF-I) in female rainbow trout. Gen. Comp. Endocrinol. 155, 257270.

695

Tsai, I.P., Madsen, S.S., Stephen, D.M., Bern, A Howard, 1994. Endocrine control of
cartilage growth in coho salmon: GH inuence in vivo on the response to IGF-I
in vitro. Zool. Sci. 11, 299303.
Uchida, K., Kajimura, S., Riley, L.G., Hirano, T., Aida, K., Grau, E.G., 2003. Effects of
fasting on growth hormone/insulin-like growth factor-I axis in the tilapia,
Oreochromis mossambicus. Comp. Biochem. Physiol. A 134, 429439.
VeraCruz, E.M., Brown, C.L., Luckenbach, J.A., Picha, M.E., Bolivar, R.B., Borski, R.J,
2006. Insulin-like growth factor-I cDNA cloning, gene expression and potential
use as a growth rate indicator in Nile tilapia, Oreochromis niloticus. Aquaculture 251, 585595.
Wilkinson, R.J., Porter, M., Woolcott, H., Longland, R., Carragher, J.F., 2006. Effects
of aquaculture related stressors and nutritional restriction on circulating
growth factors (GH, IGF-I and IGF-II) in Atlantic salmon and rainbow trout.
Comp. Biochem. Physiol. A 145, 214224.
Zhang, J.Z., Zhou, F., Wang, L.L., Shao, Q.J., Xu, Z.R., Xu, J.Z, 2010. Dietary protein
requirement of juvenile Black Sea Bream, Sparus macrocephalus. J. World
Aquacult. Soc. 41, 151164.
Zheng, K., Liang, M., Yao, H., Wang, J., Chang, Q., 2011. Effect of dietary sh protein
hydrolysate on growth, feed utilization and IGF-I levels of Japanese ounder
(Paralichthys olivaceus). Aquaculture Nutr. 4, 17.