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Fish Sci (2009) 75:967973

DOI 10.1007/s12562-009-0106-0

ORIGINAL ARTICLE

Aquaculture

Utilization of genetically modified soybean meal


in Nile tilapia Oreochromis niloticus diets
Indra Suharman Shuichi Satoh Yutaka Haga Toshio Takeuchi
Masato Endo Ikuo Hirono Takashi Aoki

Received: 30 October 2008 / Accepted: 16 March 2009 / Published online: 14 May 2009
The Japanese Society of Fisheries Science 2009

Abstract The utilization of genetically modified soybean


meal (GM SBM) was compared with that of non-GM SBM in
Nile tilapia. Four experimental diets were formulated to
include either non-GM or GM SBM at 34 or 48%, respectively. These diets were fed to juvenile Nile tilapia (49.5 g
average weight) for 12 weeks. The uptake of the cauliflower
mosaic virus 35S promoter fragment of the GM SBM in fish
muscle was examined at 8th and 12th week. After 12th week,
fish were fed the non-GM SBM diets to determine the
residual span of the incorporated promoter fragment. There
was no significant difference in specific growth rate or feed
efficiency between GM and non-GM groups at the same
inclusion level. A small number of muscles from fish
receiving both levels of GM SBM diet were positive for the
promoter fragment. Additionally, the promoter fragment
was not detected by the second day after changing to the nonGM SBM diets. These results indicate that the utilization of
GM SBM was similar to that of non-GM SBM and the promoter fragment was rarely found in fish muscles, suggesting
that suitability and safety of GM SBM in Nile tilapia diet
were similar to those of non-GM SBM.
Keywords Diets  Feed efficiency  Genetically modified
soybean  Growth  Nile tilapia  Utilization

I. Suharman  S. Satoh (&)  Y. Haga  T. Takeuchi 


M. Endo  I. Hirono  T. Aoki
Department of Marine Biosciences, Tokyo University of Marine
Science and Technology, 4-5-7 Konan, Minato, Tokyo
108-8477, Japan
e-mail: ssatoh@kaiyodai.ac.jp
I. Suharman
Department of Aquaculture, Faculty of Fisheries and Marine
Sciences, The University of Riau, Pekanbaru 28293, Indonesia

Introduction
The rapid development of global aquaculture has resulted
in increased production of aquafeeds, which traditionally
relies on fish meal as the main protein source [1]. However,
availability of fish meal is becoming limited due to
increasing demand and decreasing material fish capture.
Therefore, it is essential to evaluate the suitability of
alternative protein sources to replace fish meal as dietary
protein source in aquafeeds. Genetically modified (GM)
soybeans are becoming an increasing part of animal feed
ingredients. GM soybeans are produced by the introduction
of a gene coding for 5-enolpyruvylshikimate-3-phosphate
synthase from Agrobacterium sp. strain CP4 (CP4 EPSPS)
that confers tolerance to the herbicide glyphosate [2].
These soybeans contain recombinant gene composed of the
cauliflower mosaic virus promoter that regulates the CP4
EPSPS and the terminator of the nopaline synthase (NOS)
genes, and has received approval for commercial planting
[2], which has been gaining wide acceptance by
farmers [3].
Several studies have been conducted to determine
nutritional value and suitability of dietary GM soybeans
since soybean meal (SBM) is a preferred protein source for
various animal feeds [47]. Feeding experiments with GM
soybeans have been conducted in some species of animals
including swine [810], catfish [11], Atlantic salmon [12],
and rainbow trout [13]. From those studies, it is concluded
that GM SBM is substantially equivalent to non-GM SBM.
There were no effects on growth performance for those
animals associated with the feeds including GM SBM
compared with non-GM SBM.
Concern has been expressed about the use of GM SBM
in animal feeds, including for fish feeds. One issue raised is
whether foreign deoxyribonucleic acid (DNA) included in

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GM soybeans could be transferred to and accumulate in fish


muscles and questions regarding the safety of the fish products for human consumption. A number of studies have been
conducted to investigate the usefulness of dietary GM SBM
in diet for carnivorous fish species, such as Atlantic salmon
[14] and rainbow trout [13]. Sanden et al. [14] reported that
no foreign DNA fragment was detected in the muscle of
Atlantic salmon fed a GM SBM diet. However, other study
has shown that the cauliflower mosaic virus 35S (CaMV
35S) promoter fragment was detected in the muscle of
rainbow trout fed a GM SBM diet, but it was not present at the
second day after changing to a non-GM SBM diet [13].
Nile tilapia Oreochromis niloticus is one of the most
valuable freshwater fish and cultivated fish species of the
world [15]. This fish is an omnivorous species, utilizes a wide
spectrum of foods, and has digestive systems whose function
and morphology differ from those of carnivorous fish. In
addition, omnivorous fish are known to use dietary carbohydrate more efficiently than carnivorous fish [16]. Therefore, the objective of this study was to evaluate GM SBM as
an ingredient in diets for Nile tilapia and compare it with nonGM SBM. In addition, the fate of the CaMV 35S promoter
fragment in fish muscles fed GM SBM diet was examined.

Materials and methods


Experimental diets
Four experimental diets were formulated to include two
proportions of either non-GM SBM or GM SBM with
approximately 38% crude protein (Tables 1, 2). The diets
were pelleted using a laboratory pelletizer (AEZ12 M,
Hiraga-Seisakusho, Kobe, Japan), dried in a vacuum
freeze-drier (RLE-206, Kyowa Vacuum Tech., Saitama,
Japan), and stored at 4C until use. The chemical compositions of the experimental diets were determined as follows: crude protein using a Kjeltec Auto Sampler System
2400 Analyzer (Foss Ltd., Tokyo, Japan), crude lipid based
on the method of Folch et al. [17], and crude ash and
moisture by the procedures of Takeuchi [18].
Fish, rearing conditions, and feeding
Nile tilapia Oreochromis niloticus juveniles were obtained
from Laboratory of Fish Culture, Tokyo University of
Marine Science and Technology. Prior to the experiment, all
fish were acclimatized to the experimental condition by
feeding a commercial diet for a week. A total of 200 fish
(mean weight SE, 49.5 0.10 g) was randomly assigned
to eight 60-l glass tanks with 25 fish each. Four experimental
diets were randomly assigned to the tanks with duplicates.
Fish were fed twice daily at approximately 09:00 and 16:00 h

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Fish Sci (2009) 75:967973


Table 1 Proximate composition of soybean meal (%)
Crude protein

Crude lipid

Crude ash

Moisture

Non-GM SBM

45.4

1.8

8.4

12.5

GM SBM

44.2

3.1

5.9

12.2

Table 2 Formulation and proximate composition of the experimental


diets (%)
Diet

Non-GM SBM

GM SBM

34%

48%

34%

30

20

48%

Ingredients
Jack mackerel meal

30

20

34

48

7.9

7.9

7.9

10

10

10

GM SBM
Non-GM SBM

34

48

Wheat flour

7.9

Pregelatinized starch

10

Palm oil

Soybean oil

Vitamin premixa

Mineral premixb

Ca(H2PO4)2

Cellulose

Vitamin E (50%)

0.1

0.1

0.1

0.1

Moisture

3.9

3.6

2.3

1.9

Crude protein

38.7

39.0

38.9

38.9

Crude lipid

10.8

10.4

11.0

10.9

Crude ash

9.4

8.9

8.4

7.9

Proximate composition

The vitamin mix had the following components (mg 100 g-1):
thiamin hydrochloride 6, riboflavin 10, pyridoxine hydrochloride 4,
cyanocobalamin 0.01, ascorbic acid 500, niacin 40, Ca-pantothenate,
10, inositol 200, biotin 0.6, folic acid 1.5, p-aminobenzoic acid 5,
vitamin K3 5, vitamin A acetate 4,000 IU, vitamin D3 4,000 IU

P-free mineral mixture (g 100 g-1): NaCl 5.0, MgSO4  7H2O 74.5,
FeC6H5O7nH2O 12.5, trace element mix 5.0, cellulose 3.0. Trace
element mix compositions (mg g-1): ZnSO47H2O 353,
MnSO45H2O 162, CuSO45H2O 31, AlCl36H2O 10, CoCl6H2O 1,
KIO3 3, cellulose 440

to apparent satiation for 12 weeks. After that, fish fed with


diets containing GM SBM were fed with both inclusion
levels of the non-GM SBM diets to apparent satiation for
8 days. Fish were kept in a flow-through system with a flow
rate of 0.61.0 l/min, and sufficient aeration was supplied.
Throughout the experiment, water temperature was maintained at 22.024.0C.
Sampling and biological analysis
At the commencement of the feeding experiment, ten fish
from the stock were randomly sampled and anaesthetized
with ethylene glycol monophenyl ether (300 mg/l) before

Fish Sci (2009) 75:967973

being sacrificed; approximately 25 g dorsal muscle was


dissected from the right side of body part and immediately
frozen on dry ice and stored at -80C until DNA analysis.
The other resected tissues from ten individual fish were
pooled, homogenized, and frozen at -20C for proximate
composition analysis. Moisture, crude ash, crude protein,
and crude lipid content in the samples were determined as
described in Experimental diets. At the end of eighth
week, 20 fish were randomly taken from each treatment
and dorsal muscle samples were dissected for DNA
extraction. Moreover, at the end of 12th week, 30 fish were
weighed from each treatment for final body weight. Ten of
these fish were sacrificed and dorsal muscle samples from
the right side of body part were dissected for DNA
extraction. Those fish were pooled, homogenized, and
frozen at -20C for whole-body composition analysis.
Furthermore, dorsal muscle samples from the right side of
the body part were sampled (n = 5) at the second, fifth,
and eighth day after changing to the non-GM SBM diets.
DNA extraction
Deoxyribonucleic acid extraction from the experimental
diets was done according to Murray and Thompson [19]
with minor modifications: 1 ml CTAB (hexadecyltrimethyl-ammonium bromide) buffer (20 g/l CTAB,
0.1 M TrisHCl pH 8.0, 1.4 M NaCl, and 20 mM EDTA)
was added to 0.1 g sample, mixed by vortex, and incubated
in a water bath at 60C for 60 min. Then, the solution was
centrifuged at 21,8409g for 3 min at 4C. The supernatant
was transferred to a new tube, filled with 200 ll phenol:chloroform:isoamyl alcohol (PCI, 25:24:1), mixed
gently by inverting for 5 min, and centrifuged at 21,8409g
for 3 min at 4C. The upper layer was transferred to a 1.5ml tube and 200 ll PCI was added, mixed gently by
inverting for 20 min, and centrifuged at 21,8409g for
10 min at 4C. The upper layer was gently mixed with
300 ll 2-propanol for 30 s, and centrifuged at 21,8409g
for 5 min at 4C. The DNA pellet was washed with 800 ll
75% ethanol, centrifuged at 21,8409g for 5 min at 4C,
and dried at room temperature. The DNA pellet was dissolved in 50 ll TE buffer (10 mM TrisHCl, 1 mM
EDTA, pH 8.0) as a stock DNA solution, and stored at
-20C. DNA was extracted from fish muscle sample
(approximately 0.25 g) using the phenolchloroform
method. The sample was digested with 40 lg proteinase K
in 400 ll TNESurea buffer (10 mM TrisHCl pH 7.5,
125 mM NaCl, 10 mM EDTA, 0.5% SDS, 5 M urea)
overnight at 37C. DNA was extracted with 400 ll PCI for
20 min over a rotary shaker (NV-220, Nissin, Tokyo,
Japan), and centrifuged at 21,8409g for 10 min at room
temperature. The upper layer was transferred to a 1.5-ml
tube and 300 ll PCI was added, mixed gently for 20 min

969

by a rotary shaker, and then centrifuged at 21,8409g for


10 min at room temperature. To the upper layer was added
800 ll absolute ethanol (-20C) for precipitating, followed by centrifugation at 21,8409g for 10 min at room
temperature. The DNA pellet was washed twice with
400 ll 70% ethanol (-20C) and centrifuged at 21,8409g
for 5 min at room temperature. The DNA pellet was airdried, dissolved in 30 ll TE buffer, and stored at -20C.
Polymerase chain reaction
One pair of specific primers was designed and used to
detect the CaMV 35S fragment that is present in the
genomic structure of GM soybeans. The primer pair 35S-F
(50 -CCT ACA AAT GCC ATC ATT GCG-30 ) and 35S-R
(50 -GGG TCT TGC GAA GGA TAG TG-30 ) were
designed against the sequence of the CaMV 35S promoter
(GenBank accession no. V00141) to amplify a 205-bp
fragment. Polymerase chain reaction (PCR) was carried out
in a reaction mixture (20 ll) containing 2 ll 109 Taq
buffer with 20 mM MgCl2, 1.5 ll dNTP mixture, 1 ll
primers, 0.05 ll Taq polymerase (Takara, Japan), 14.45 ll
distilled water, and 1 ll DNA template. The PCR conditions were: 94C for 10 min (initial denaturation), 40
cycles of 95C for 25 s (denaturation), 60C for 30 s
(annealing), and 72C for 45 s (extension), followed by
72C for 7 min (final extension). The PCR products were
separated on a 1.5% (w/v) agarose gel containing 1%
ethidium bromide. Electrophoresis was carried out at
approximately 100 V for 3545 min, visualized under
ultraviolet (UV) light at 254 nm and photographed using a
charge-coupled device (CCD) camera (SFC Instruments,
Japan).
DNA sequencing
The PCR product was labeled with Thermo Sequenase
Cycle Sequencing Kit (Amersham Biosciences) for
sequencing using the CaMV 35S primers. This PCR
product was sequenced by DNA Sequencer (Applied Biosystems 3130xl Genetic Analyzer, Hitachi, USA).
Sequence data was then pasted into the BLAST test similarity of sequence within the database.
Statistical analysis
Data on fish and feed performance were statistically evaluated by Statistica 6 (Statsoft, USA, 2001). All data were
subjected to one-way analysis of variance (ANOVA), and
Tukeys honest significant difference (HSD) test was used
to determine if the differences among means were statistically significant. Differences between means were considered significant at P \ 0.05.

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Fish Sci (2009) 75:967973

Table 3 Growth and feed performance in Nile tilapia fed diets with different levels of non-GM and GM SBM for 12 weeks (mean standard
error, SE)
Treatments
Performance parameters

Experimental diets
Non-GM SBM 34%

Non-GM SBM 48%

GM SBM 34%

GM SBM 48%

Initial body weight (g)

49.5 0.10

49.2 0.40

49.2 0.44

50.2 0.24

Final body weight (g)

165 8.71

136 1.81

168 12.8

168 1.75

WGa
SGRb

115 8.81
1.43 0.07

87.1 1.40
1.21 0.01

119 12.4
1.46 0.08

118 1.51
1.44 0.01

FEc

0.71 0.00

0.71 0.03

0.75 0.04

0.66 0.01

3.63 0.02

3.59 0.15

3.44 0.16

3.92 0.08

57.5 2.59

56.5 2.99

58.8 2.06

59.8 1.68

PER

PRe
a

Weight gain (WG) (g) = mean final weight (FW) (g) - mean initial weight (IW) (g)

Specific growth rate (SGR) (% body weight/day) = [(ln FW (g) ln IW (g))/duration of feeding trial (days)] 9 100

Feed efficiency (FE) (g/g) = WG (g)/feed intake (g)

Protein efficiency ratio (PER) (g/g) = WG (g)/protein intake (g)

Protein retention (PR) (%) = [(FW 9 final body protein) - (IW 9 initial body protein)]/(feed intake 9 feed protein) 9 100

Results

Table 4 Whole-body composition (% wet weight) of Nile tilapia fed


diets with different levels of non-GM or GM SBM for 12 weeks

Composition of soybean meals and experimental diets


Proximate composition of GM SBM was comparable to
that of non-GM SBM (Table 1). The GM SBM had slightly
higher crude lipid and lower crude ash content than those
of non-GM SBM. Proximate composition of the experimental diets was approximately 38.739.0% of crude
protein, 10.411.0% of crude lipid, 7.99.4% of crude ash,
and 1.93.9% of moisture (Table 2).

Crude
protein
Initial

Crude
lipid

Crude ash Moisture

16.3 0.1 7.3 0.0 4.5 0.0

70.2 0.4

Non-GM SBM 34% 16.0 0.4 6.9 0.4 4.2 0.1b 72.0 1.9
Non-GM SBM 48% 15.9 0.1 6.4 0.5 4.4 0.2b 71.5 1.1
GM SBM 34%
16.1 0.1 8.3 0.7 4.1 0.1b 72.2 0.3
GM SBM 48%

15.6 0.1 8.8 0.3 2.5 0.1a 71.8 0.3

Values (mean SE) in the same column with the same superscript
are not significantly different (P [ 0.05)

Growth performance and feed utilization


All fish readily accepted the experimental diets and no fish
died during the 12-week feeding trial. Weight gain (WG)
and specific growth rate (SGR) were not significantly different in groups receiving either non-GM or GM SBM at
different inclusion levels (Table 3). There were no significant differences in feed efficiency (FE) among all the
treatments. There were no significant differences in protein
efficiency ratio (PER) between non-GM and GM SBM
groups at the same inclusion level. There were no significant differences in protein retention (PR) between non-GM
or GM SBM groups.
Whole-body composition
There were no significant differences in moisture and crude
protein content in the whole body of Nile tilapia (Table 4),
although crude protein content decreased slightly with
increasing soybean meal inclusion in the experimental
diets. Crude lipid content in the whole body of fish fed GM
SBM diets was slightly higher compared with those of fish

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fed non-GM SBM diets. Fish from all experimental groups


showed higher moisture content after 12-week feeding trial
compared with the initial fish.
Detection of the CaMV 35S promoter fragment
The CaMV 35S promoter fragment was detected in the GM
SBM, but not in the non-GM SBM counterpart. In addition,
strong positive signal of the CaMV 35S promoter fragment
was detected only in the GM SBM diets (data not shown).
This indicates that cross-contamination between non-GM
and GM SBM during preparation of the experimental diets
had been excluded.
PCR was applied to examine the existence of the CaMV
35S promoter fragment in the muscle of fish fed non-GM
or GM SBM diets. The result showed that the CaMV 35S
promoter fragment was not detected in the muscle samples
of fish fed diets with two different inclusion levels of nonGM SBM. However, the fragment was detected in 2 fish
muscle samples from 20 samples from fish fed 48% GM

Fish Sci (2009) 75:967973


Fig. 1 Detection of CaMV 35S
promoter fragments in muscles
of Nile tilapia fed with non-GM
or GM SBM diets at the end of
eighth week. M 100-bp DNA
marker, - control without
DNA, ? positive control using
GM SBM DNA, 120
individual Nile tilapia samples

971

Non-GM SBM 34%


Non-GM SBM 48%

GM SBM 34%

GM SBM 48%
M

1 2

3 4 5 6 7 8

10 11 12 13 14 15 16 17 18 19 20

Table 5 Number of detectable fragment of CaMV 35S promoter in muscles from fish fed non-GM or GM SBM diets at 8 and 12 weeks, and
after changing to non-GM SBM diet
Sampling day (weeks)/fish number sampleda

Fed with non-GM or GM SBM diets

Fed with non-GM SBM diet

56 (8)/20

84 (12)/10

86/5

89/5

92/5

Non-GM SBM 34%

Non-GM SBM 48%

GM SBM 34%
GM SBM 48%

0
2

1
0

0
0

0
0

0
0

Number of fish/treatment

SBM diet at the end of eighth week (Fig. 1). In addition,


only one sample was positive out of ten in the muscle
samples from fish fed 34% GM SBM diet at the end of 12th
week (Table 5). Furthermore, the CaMV 35S promoter
fragment was not detected in the muscle samples of fish fed
both GM SBM diets after changing to the non-GM SBM
diets (Table 5). The identity of the CaMV 35S PCR
product of fish muscle sample was verified by sequence
analysis, showing it to be identical to the CaMV 35S
promoter reference sequence (GenBank accession no.
V00141) (data not shown).

Discussion
In the present study, growth performance and feed utilization of Nile tilapia fed GM SBM diet were similar to
those of fish fed on non-GM SBM diet. These results are in
agreement with previous studies on other species such as
catfish [11], Atlantic salmon [12, 14, 20, 21], and rainbow
trout [13]. Similar results were also reported in swine [8],
rat [22], and broiler [23]. This would be caused by similar
feed compositions between GM and non-GM soybean diets
given to the experimental fish. These data suggest that GM
SBM is equivalent to non-GM SBM based on growth of
Nile tilapia. From these findings, it seems that omnivorous
fish fed either GM SBM or non-GM SBM diet was not
significantly different from carnivorous fish in term of
growth and feed utilization.

The present results showed no difference in the chemical


composition of GM SBM and non-GM SBM. Previous
studies have suggested that no compositional differences
exist between GM and conventional soybeans [5, 8, 22,
23]. Based on these findings, it is concluded that SBM
prepared from GM herbicide-tolerant soybean is equivalent
to non-GM SBM with respect to chemical composition.
Concerning proximate composition, crude protein content of Nile tilapia was not greatly affected by inclusion
level of SBM. These results agree with findings of Pongmaneerat et al. [24] and El-Saidy and Gaber [25]. Also,
whole-body lipid and moisture contents did not vary significantly when compared with those of the non-GM
groups. Thus, it is concluded that utilization of GM SBM
for Nile tilapia would be comparable to that of non-GM
SBM.
Examination of the CaMV 35S promoter residue in fish
muscle revealed that most of the muscle samples taken
from fish fed GM SBM diets were negative for the 205-bp
fragment of the CaMV 35S promoter. This result is consistent with the previous study on Atlantic salmon by
Sanden et al. [14], who reported that GM soy fragment
(195 bp) was not detected in the muscle of Atlantic salmon
fed a GM SBM-based diet. Ash et al. [26] also reported
that no transgenic DNA was found in the tissues of hens
fed GM SBM. Similar results were observed in broiler and
pig fed GM corn [2729]. We assume that most of foreign
DNA was rapidly degraded to fragments smaller than
180 bp in the digestive tract, being below the level of

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detection. In addition, Nile tilapia has high capacity to


digest dietary plant protein [30]. This ability is attributed to
the very low stomach pH in this species, which allows them
to digest and extract the cellular content even without
breaking the cellulose cell wall [31].
However, a small number of fish muscle samples were
positive for the CaMV 35S promoter. One possible
explanation would be attributed to intra-individual difference; the foreign DNA is rather slowly degraded by these
fish in the gastrointestinal (GI) tract. Mazza et al. [32]
suggested that the DNA fragments are rapidly degraded
before reaching peripheral organs. Another possibility was
the presence of blood in fish muscle. The remaining DNA
in the GI tract might be absorbed and transported by blood
circulation. Thus, it is possible for the CaMV promoter
fragments to be detected in the muscle as a result of the
presence of blood in this tissue [33].
The present results demonstrate that utilization of protein in GM SBM is similar to that of non-GM SBM with no
negative effects on growth and feed efficiency. The promoter fragment was rarely detected in muscles of fish fed
GM SBM diets and was not present by the second day after
changing to the non-GM SBM diets, verifying the suitability of GM SBM in Nile tilapia diets.

Fish Sci (2009) 75:967973

9.

10.

11.

12.

13.

14.

15.

16.
Acknowledgments The authors are grateful to the Japanese Ministry of Education, Culture, Sports, Science, and Technology (MEXT)
for the scholarship grant extended to Indra Suharman. This study was
financially supported in part by a Grant-in-Aid for Scientific Research
from Japan MEXT (B, 19380121) to Shuichi Satoh.

References

17.

18.

19.
20.

1. Tacon AGJ, Jackson AJ (1985) Utilization of conventional and


unconventional protein sources in practical fish feeds. In: Cowey
CB, Mackie AM, Bell JG (eds) Nutrition and feeding of fish.
Academic Press, London, pp 119145
2. Padgette SR, Kolacz KH, Delannay X, Re DB, Lavallee BJ, Tinius CN, Rhodes WK, Otero YI, Barry GF, Eichholtz DA, Peschke VM, Nida DL, Taylor NB, Kishore GM (1995)
Development, identification and characterization of a glyphosatetolerant soybean line. Crop Sci 35:14511461
3. Swick RA (2000) Feeding genetically enhanced soy to animals.
ASA Tech Bull AN 28:19
4. Burks AW, Fuchs RL (1995) Assessment of the endogenous
allergens in glyphosate-tolerant and commercial soybean varieties. J Allergy Clin Immunol 96:10081010
5. Padgette SR, Taylor NB, Nida DL, Bailey MR, Macdonald J,
Holden LR, Fuchs RL (1996) The composition of glyphosatetolerant soybean seeds is equivalent to that of conventional
soybeans. J Nutr 126:702716
6. Astwood JD, Leach JN, Fuchs RL (1996) Stability of food
allergens to digestion in vitro. Nat Biotechnol 14:12691273
7. Nelson KA, Renner KA (1999) Weed management in wide- and
narrow-row glyphosate-resistant soybean. J Prod Agric 12:460
465
8. Cromwell GL, Lindemann MD, Randolph JH, Parker GR, Coffey
RD, Laurent KM, Armstrong CL, Mikel WB, Stanisiewski EP,

123

21.

22.

23.

24.

25.

26.

Hartnell GF (2002) Soybean meal from roundup ready or conventional soybeans in diets for growing-finishing swine. J Anim
Sci 80:708715
Aumaitre A, Aulrich K, Chesson A, Flachowsky G, Piva G
(2002) New feeds from genetically modified plants: substantial
equivalence, nutritional equivalence, digestibility and safety for
animals and the food chain. Livest Prod Sci 74:223228
Faust MA (2002) New feeds from genetically modified plants: the
US approach to safety for animals and the food chain. Livest Prod
Sci 74:239254
Hammond B, Vicini JL, Hartnell GF, Naylor MW, Knight CD,
Robinson EH, Fuchs RL, Padgette SR (1996) The feeding value
of soybeans fed to rats, chickens, catfish and dairy cattle is not
altered by genetic incorporation of glyphosate tolerance. J Nutr
126:717727
Hemre GI, Sanden M, Bakke-McKellep MA, Sagstad A, Krogdahl
A (2005) Growth feed utilization and health of Atlantic salmon
(Salmo salar L.) fed genetically modified compared to non-modified commercial hybrid soybeans. Aquac Nutr 11:157167
Chainark P, Satoh S, Tokuya H, Viswanath K, Hirono I, Aoki T
(2006) Availability of genetically modified soybean meal in rainbow trout (Oncorhynchus mykiss) diets. Fish Sci 72:10721078
Sanden M, Bruce IJ, Rahman MA, Hemre G (2004) The fate of
transgenic sequences present in genetically modified plant products in fish feed, investigating the survival of GM soybean DNA
fragments during feeding trials in Atlantic salmon, Salmo salar L.
Aquaculture 237:391405
El-Sayed AFM (1998) Total replacement of fish meal with animal
protein sources in Nile tilapia, Oreochromis niloticus (L.), feeds.
Aquac Res 29:275280
Wilson RP (1994) Utilization of dietary carbohydrate by fish.
Aquaculture 124:6780
Folch J, Lee M, Stanley GHS (1957) A simple method for the
isolation and purification of total lipids from animal tissues. J
Biol Chem 226:497507
Takeuchi T (1988) Chemical evaluation of dietary nutrients. In:
Watanabe T (ed) Fish nutrition and mariculture. Japan International Cooperation Agency (JICA), Kanazawa, pp 79228
Murray M, Thompson W (1980) Rapid isolation of high-molecular-weight plant DNA. Nucleic Acids Res 8:43214325
Sanden M, Berntssen MH, Krogdahl A, Hemre GI, BakkeMckellep AM (2005) An examination of the intestinal tract of
Atlantic salmon, Salmo salar L. parr fed different varieties of soy
and maize. J Fish Dis 28:317330
Sanden M, Krogdahl A, Bakke-Mckellep AM, Buddington RK,
Hemre GI (2006) Growth performance and organ development in
Atlantic salmon, Salmo salar L. parr fed genetically modified
(GM) soybean and maize. Aquac Nutr 12:112
Zhu Y, Li D, Wang F, Yin J, Jin H (2004) Nutritional assessment
and fate of DNA of soybean meal from roundup ready or conventional soybeans using rats. Arch Anim Nutr 58:295310
Taylor M, Hartnell G, Lucas D, Davis S, Nemeth M (2007)
Comparison of broiler performance and carcass parameters when
fed diets containing soybean meal produced from glyphosatetolerant (MON 89788), control, or conventional reference soybeans. Poult Sci 86:26082614
Pongmaneerat J, Watanabe T, Takeuchi T, Satoh S (1993) Use of
different protein meals as partial or total substitution for fish meal
in carp diets. Nippon Suisan Gakkaishi 59:12491257
El-Saidy DMS, Gaber MMA (2003) Replacement of fish meal
with a mixture of different plant protein sources in juvenile Nile
tilapia Oreochromis niloticus (L.) diets. Aquac Res 34:1119
1127
Ash J, Novak C, Scheideler SE (2003) The fate of genetically
modified protein from roundup ready soybeans in laying hens.
J Appl Poult Res 12:242245

Fish Sci (2009) 75:967973


27. Yonemochi C, Fujisaki H, Harada C, Kusama T, Hanazumi M
(2002) Evaluation of transgenic event CBH 351 (StarLink) corn
in broiler chicks. J Anim Sci 73:221228
28. Jennings JC, Albee LD, Kolwyck DC, Surber JB, Taylor ML,
Hartnell GF, Lirette RP, Glenn KC (2003) Attempts to detect
transgenic and endogenous plant DNA and transgenic protein in
muscle from broilers fed yieldgard corn borer corn. Poult Sci
82:371380
29. Chowdhury EH, Kuribara H, Hino A, Sultana P, Mikami O,
Shimada N, Guruge KS, Saito M, Nakajima Y (2003) Detection
of corn intrinsic and recombinant DNA fragments and Cry1Ab
protein in the gastrointestinal contents of pigs fed genetically
modified corn Bt11. J Anim Sci 81:25462551

973
30. Watanabe T, Takeuchi T, Satoh S, Kiron V (1996) Digestible
crude protein contents in various feedstuffs determined with four
freshwater fish species. Fish Sci 62:278282
31. Bowen SH (1981) Digestion and assimilation of periphytic
detrital aggregate by Tilapia mossambica. Trans Am Fish Soc
110:239245
32. Mazza R, Soave M, Morlacchini M, Piva G, Marocco A (2005)
Assessing the transfer of genetically modified DNA from feed to
animal tissues. Transgenic Res 14:775784
33. Chainark P, Satoh S, Hirono I, Aoki T, Endo M (2008) Availability of genetically modified feed ingredient: investigations of
ingested foreign DNA in rainbow trout Oncorhynchus mykiss.
Fish Sci 78:380390

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