- Email: [email protected]
Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: Economic, zootechnical and nutritional meat improvements
Accepted Manuscript Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: Economic, zootechnical and nutritional meat improvements
Antonio Cesar Godoy, Oscar Oliveira Santos, Jarred Hugh Oxford, Iury Walysson de Amorim Melo, Rômulo Batista Rodrigues, Dacley Neu, Ricardo Nunes, Wilson Rogério Boscolo PII: DOI: Article Number: Reference:
S0044-8486(19)30467-3 https://doi.org/10.1016/j.aquaculture.2019.734324 734324 AQUA 734324
To appear in:
aquaculture
Received date: Revised date: Accepted date:
23 February 2019 15 June 2019 18 July 2019
Please cite this article as: A.C. Godoy, O.O. Santos, J.H. Oxford, et al., Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: Economic, zootechnical and nutritional meat improvements, aquaculture, https://doi.org/10.1016/ j.aquaculture.2019.734324
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: economic, zootechnical and nutritional meat improvements Antonio Cesar Godoya,*, Oscar Oliveira Santosa, Jarred Hugh Oxfordb, Iury Walysson de Amorim
Universidade Estadual de Maringá. Av. Colombo, 5790 - Centro de Ciências Exatas -
IP
a
T
Meloc, Rômulo Batista Rodriguesc, Dacley Neuc, Ricardo Nunesc, Wilson Rogério Boscolo c,*
b c
CR
Departamento de Química. Maringá/PR, Brazil
University of Georgia - Poultry Science Department. Athens GA. USA
Universidade Estadual do Oeste do Paraná. Centro de Engenharias e Ciências Exatas. Rua da
US
Faculdade, 645, Toledo/PR, Brazil.
AN
Abstract
The aim of this study was to evaluate different levels of soybean oil inclusion in diets for Nile
M
tilapia. 270 tilapia, with an initial weight of 425.33 ± 32.37 g and initial length of 25.53 ± 2.00 cm, were randomly distributed in 15 tanks. The diets were formulated to be isonitrogenous (30%)
ED
using five increasing levels of soybean oil (0.00; 15.00; 30.00; 45.00 and 60.00 g kg −1) with each level having three replicates. The evaluated parameters were: performance (survival, daily weight
PT
gain, feed conversion, hepatosomatic index, visceral fat, carcass yield, fillet yield and dressed out yield), hematological and biochemical blood aspects, total protein, cholesterol, glucose,
CE
triglycerides, hemoglobin, hematocrit and red blood cells, and the chemical composition of the fillets (moisture, crude protein, lipids, and ash). Quadratic effects ( P < 0.05 ) were observed for
AC
the variables: daily weight, weight gain, final weight, carcass yield, visceral fat, and hepatosomatic index. For blood variables, a quadratic effect ( P < 0.05 ) for total cholesterol was observed. For chemical composition analyses of the fillets, there were no effects ( P > 0.05 ) observed with the increase of soybean oil. A total of 22 fatty acids were determined in the fillets of the tilapia fed with different soybean oil levels. Among fatty acids, the highest concentrations were for 16:0 *
Corresponding author
Email
address:
[email protected]
[email protected] (Wilson Rogério Boscolo)
(Antonio
Cesar
Godoy),
ACCEPTED MANUSCRIPT (238.40 to 262.19 mg g−1); 18:1n-9 (215.36 to 277.61) and 18:2n-6 (LA) (157.84 to 224.49). It is concluded that the addition of around 45.0 g kg−1 of soybean oil to the finisher diet of Nile tilapia provides better growth performance. Animals fed the diet, used the nutrients to produce recommended PUFA/SFA values. Dietary supplementation was also effective at improving the nutrition values of the meat.
T
Keywords: economic improvement, fish growth performance, essentials fatty acid, human
CR
1.
IP
nutrition
Introduction
US
In aquaculture one of the most prominently farmed species is the Nile tilapia (Oreochromis niloticus), this can be attributed to the rapid growth, hardiness, large fillet yield, absence of
AN
intramuscular bones, and excellent acceptance by the consumer market [30]. The tropical fish most farmed in the world [51] and it still growing [22, 23, 24] it is estimated that the production of
M
tilapia will reach approximately 6.6 million tons by 2030 [51].
ED
Fish, in general, have excellent nutritional properties, with balanced protein compositions, essential fatty acids, vitamins, and minerals. Another benefit of fish is the ability to improve the nutritional quality of the meat through supplementation with different additions of fatty acids and
PT
vitamins sources in the diet [44, 19, 56].
There are many components in fish diets with the lipids being considered as an essential
CE
energy source that can significantly influence feed conversion and fish growth [57]. In intensive fish farming, high levels of fat sources in the diet are used in order to meet the fish’s energy
AC
requirements; this is done to prevent the breakdown of protein to be used as an energy source in the body [25]. However, excessive energy supply can influence the animal’s metabolism and cause adverse effects on growth, carcass composition with undesirable fat accumulation in the viscera and abdominal cavity. As well as reducing the percentage of fillet yield and the commercial value of the fish, it has been shown that the tissue lipid composition reflects the feed and can be changed by the manipulation of the diet [35, 60, 54]. Vegetable oils are regarded as valuable energy sources for tropical fish, because they are easily found on the market, and are used according to their availability and cost of the raw
ACCEPTED MANUSCRIPT materials in each region. Soybean oil is more frequently used due to its low cost when compared to other energy sources, and its overall composition meets the animal’s nutritional demands [33]. Soybean oil has an apparent digestibility of 89.85% and 8,485 kcal kg −1 of digestible energy, demonstrating that this nutrient is well used by the species O. niloticus [8]. Some studies have evaluated the inclusion of soybean oil in diets of Nile tilapia showing good results in the literature. The addition of 5.9% of soybean oil in the diet increases the carcass
T
and fillet yields of the tilapia [10, 43]. However, few studies have evaluated the inclusion of
IP
soybean oil in the finisher phase diets of tilapia.
CR
The inclusion of lipids in fish feed might be a potential mechanism to reduce protein levels and commercial cost in tilapia diets if an appropriate level is identified [31]. Due to the small
US
number of studies on the inclusion of soybean oil in the finisher diets of tilapia, this study aimed to evaluate varying levels of soybean oil in the diet of Nile tilapia and to evaluate its effects on
AN
performance parameters, hematological parameters, fillet composition and fatty acids
M
incorporation of the Nile tilapia during the finisher phase.
Materials and methods
2.1.
Experimental design and diets
ED
2.
PT
This study was conducted at the Instituto de Pesquisa em Piscicultura Ambiental (InPAA), at Universidade Estadual do Oeste do Paraná campus Toledo-PR, Brazil, and it was approved by the
CE
ethics committee for animals use in CEUA at the same institution under protocol No. 36/14. The experiment was conducted for 50 days, in which 270 Nile tilapia (O. niloticus) with an initial average weight of 425.33 ± 32.37 g and 25.53 ± 2.00 cm of length were randomly distributed in 15
AC
tanks with dimensions 2.0x6.0x0.6 m, with a capacity of 7.2 m3. The fish were fed until apparent satiation, twice a day during the finishing phase. A completely randomized design was used with five treatments and three replications. The treatments consisted of soybean oil inclusion in the diet a (0.0; 15.0; 30.0; 45.0 and 60.0 g kg−1). The diets (Table 1 and Table 2) were formulated to contain around 303.80 g kg −1 of crude protein and from 12.92 to 14.34 MJ kg−1 of digestible energy, with estimated values based on the suggested values for Nile tilapia (O. niloticus) by the [48]. The increase in soybean oil content was made at the expense of cornmeal in the diets. The addition of vitamins and minerals occurred
ACCEPTED MANUSCRIPT through a commercial premix, which meets the requirements of the species.
Table 1: Composition and nutritional diets with different soybean oil inclusions Oreochromis niloticus. Soybean oil inclusion g kg−1 15.0
30.0
45.0
60.0
Corn
254.1
236.7
219.4
202.0
184.6
Soybean meal 46%
120.9
123.3
125.8
130.6
Wheat middling
150.0
150.0
IP
128.2
150.0
150.0
150.0
Wheat bran
169.5
169.0
168.6
167.6
Bone and meat meal 42%
24.0
24.9
25.3
25.8
26.30
Poultry by-product meal 58%
200.0
200.0
CR
168.1
200.0
200.0
200.0
Feather meal 84%
60.0
60.0
60.0
60.0
60.0
Soybean oil
0.00
15.0
30.0
45.0
60.0
Choline chloride 75%
1.50
1.50
1.50
1.50
1.50
Sodium chloride
3.00
3.00
3.00
3.00
3.00
DL-Methionine
2.70
2.70
2.70
2.80
2.80
L-lysine 50.7%
6.30
6.20
6.10
6.00
6.00
2.20
2.20
2.20
2.20
2.20
0.40
0.40
0.40
0.40
0.40
0.10
0.10
0.10
0.10
0.10
5.0
5.0
5.0
5.0
5.0
0.34
0.35
0.36
0.36
0.37
PT
Vitamin C Antioxidant 1
CE
Vitamin and mineral supplement for fish 2
AC
Cost of diets (US$ kg−1)
Moisture
US AN
ED
L-Threonine
T
0.00
M
Ingredients g kg−1
−1
Analysed values (g kg ) 76.53
76.49
77.03
76.87
76.32
Crude protein
302.49
303.89
303.18
302.98
303.67
Crude fiber
40.16
41.75
40.92
40.15
41.07
Crude fat
49.31
66.22
79.82
93.91
111.22
Ash
75.97
77.34
77.97
75.34
77.35
Calcium
12.37
13.88
14.03
12.96
13.98
Phosphorus
11.53
11.77
12.67
12.12
12.34
ACCEPTED MANUSCRIPT Calculated values (g kg−1) 18.00
18.00
18.00
18.00
18.00
DL-Methionine
7.50
7.50
7.50
7.50
7.50
Methionine + cystine
13.10
13.10
13.10
13.10
13.10
Arginine
20.30
20.30
20.30
20.30
20.30
Threonine
13.50
13.50
13.50
13.50
13.50
Tryptophan
3.10
3.10
3.10
3.10
3.10
Choline g kg−1
3.00
3.00
3.00
3.00
3.00
Fish digestible energy MJ kg−1
12.92
13.25
13.96
14.34
Fish digestible protein
262.00
262.00
262.00
262.00
262.00
Starch
296.20
284.00
271.80
259.60
247.50
IP
CR
1
13.63
BHT (butylated hydroxytoluene).
2
US
*All ingredients were bought at local market.
T
Lysine
Supplementation levels per kg of feed: Vit. A 12,000 IU; Vit. D3 3000 IU; Vit. K3 15 mg kg −1; Vit.
AN
B1 20 mg kgVit. B2 20 mg kg−1; Vit. B6 18 mg kg−1; Vit. B12 0.04 mg kg−1 ; Vit C 300 mg kg−1; Niacin 100 mg kg−1; Calcium Pantothenate 50 mg kg−1; Biotin 1 mg kg−1; Folic Acid 6 mg kg−1;
M
Inositol 150 mg kg−1; Choline 500 mg kg−1; Sulf. Copper 18 mg kg−1; Sulf. Iron 80 mg kg−1; Sulf. Manganese 50 mg kg−1; Sulf. Zinc 120 mg kg−1; Calcium iodate 0.8 mg kg−1; Sulf. Cobalt 0.6 mg
ED
kg−1; selenium 4 mg kg−1, Vit. E 200 mg kg−1. 3 Based on available nutrients (NRC, 2011). and
PT
calculated by Super Crac Software (TD Software, Viçosa, MG, Brazil.
Fatty acid (mg g−1)
0.00
Soybean oil inclusion g kg−1 15.0
30.0
60.0
2.61 ±0.08
205.51 ±4.41
115.96 ±2.41
64.88 ±1.41
61.16 ±1.35
61.44 ±1.36
63.33 ±1.41
62.88 ±1.39
16:1n-9
0.69 ±0.03
0.30 ±0.02
0.20 ±0.01
0.13 ±0.01
0.13 ±0.01
16:1n-7
1.75 ±0.09
1.41 ±0.07
1.49 ±0.08
1.49 ±0.08
1.46 ±0.08
18:1n-9
171.52 ±4.32
207.00 ±5.22
203.26 ±5.21 204.65 ±5.16
207.99 ±5.24
18:1n-7
33.51 ±1.18
23.54 ±0.82
18:2n-6
295.3 ±6.94
361.47 ±8.51
16:0 18:0
2.48 ±0.08
45.0
5.48 ±0.18
AC
14:0
CE
Table 2: Fatty acid composition (mg g−1) of total lipids of experimental diets.
2.45 ±0.08
2.34 ±0.08
116.35 ±2.44 116.37 ±2.44
116.45 ±2.44
23.82 ±0.84
11.27 ±0.40
11.36 ±0.41
362.99 ±8.54 368.48 ±8.67
378.71 ±8.91
ACCEPTED MANUSCRIPT 18:3n-3
27.58 ±0.34
39.20 ±0.48
41.56 ±0.51
43.45 ±0.53
44.95 ±0.55
n 6 / n3
10.71 ±0.14
9.22 ±0.11
8.73 ±0.11
8.48 ±0.10
8.43 ±0.10
SFA
275.87 ±5.81
179.73 ±3.81
180.27 ±3.83 182.15 ±3.87
181.67 ±3.86
MUFA
207.47 ±4.59
232.24 ±5.14
228.77 ±5.06 217.53 ±4.81
220.95 ±4.88
PUFA
322.88 ±7.50
400.67 ±9.30
404.55 ±9.39 411.93 ±9.57
423.66 ±9.84
806.22 ±12.28
812.63 ±12.37
813.58 ±12.39 811.6 ±12.36 826.27±12.59
T
IP
*Results expressed as mean ± standard deviation of three replicates; values with different letters on the same line are significantly different (Tukey) P < 0.05 ; LA: linoleic acid; AA: arachidonic
CR
acid; LNA: -linolenic acid; EPA: eicosapentaenoic acid; DHA: docosahexaenoic acid; SFA: total saturated fatty acids; MUFA: total of monounsaturated fatty acids; PUFA: total
US
polyunsaturated fatty acids; n-6: total of omega 6 fatty acids; n-3: total omega-3 fatty acids; n-6/n-3: rate of omega 6/omega 3; PUFA/SFA: total polyunsaturated fatty acids/total saturated
AN
fatty acids.
M
For the development of diets, the feeds were individually milled in a hammer mill (Vieira, MCS 280, Tatuí-São Paulo, Brazil) with a 0.5 mm sieve. After milling, the ingredients were
ED
weighed and mixed manually. For the inclusion of the different levels of soybean oil in the diet, a premix of the oil was made with a sample of the other ingredients and then mixed with the rest of
PT
the diet. Afterwards, the whole mixture was moistened with 22% of water and processed through the extrusion process with a 3.0 mm matrix through an extruder (Ex-Micro® extruder, ExTeec
CE
Company, Ribeirão Preto, Brazil), keeping the temperature at 80°C, o ensure the diets were not overheated. The diets were dried out in a forced air oven (ET-394/3 Tecnal Piracicaba, Brazil) for
2.2.
AC
12 hours at 55°C.
Water parameters
The physical and chemical water parameters, such as dissolved oxygen (mg L−1), pH and electrical conductivity (µS cm−1) were measured weekly, while the temperature (°C) was measured twice a day with the assistance of the YSI Professional Plus Multiparameter Water Quality Meter device (YSI Pro Plus, Yellow Springs, Ohio, USA). The physical and chemical parameters of the water were on average 7.5 ± 0.83 mg O2 L−1; 6.98 ± 0.86 and 33.42 ± 14.64 µS cm−1 for dissolved oxygen, pH and electrical conductivity, respectively.
ACCEPTED MANUSCRIPT
2.3.
Zootechnical parameters
At the end of the trial, the fish were fasted for 24 hours, to ensure gastrointestinal tract was empty, and then were euthanized with a water benzocaine solution at 250 mg L−1 [39]. The fish were weighed, measured and counted to determine the performance parameters. The weight gain (initial
T
body weight - final body weight), daily weight gain (weight gain/day), survival [(final number of
IP
fish / initial number of fish) x100] and feed conversion (dry diet feed (g) / wet weight gained (g)) were evaluated. The Diet cost rate was calculated, following DCR = diet cost rate (per kg) / weight
CR
gained. Nine fish in each experimental unit were used for the carcass yield assessments ([(body weight (g) - visceral fat weight (g))/(body weight (g))x100]), dressed out yield ([(body weight (g)
US
- (visceral weight (g) + fish head(g))) / (body weight (g)) x100]) and fillet yield ((fillet weigh (g)/body weight (g))x100), hepatosomatic index ([liver weight (g)x100 / final body weight (g)])
Biochemical parameters of blood
M
2.4.
AN
and visceral fat ([visceral fat weight (g) x100 / final body weight (g)]).
ED
Three fish were selected from each experimental unit (nine fish per treatment), for the analysis of hematological and biochemical blood parameters. The fish were anesthetized with benzocaine (100 mg L−1) following the methods described by [50]. An erythrocyte count was performed in a
PT
Neubauer chamber using the microhematocrit method according to [27], and hemoglobin by the cyanmethaemoglobin method [14]. For biochemical blood parameters, plasma was analyzed for
CE
the following parameters: plasma glucose, total serum proteins, total cholesterol, and triglycerides were evaluated using enzymatic-colorimetric methods with the aid of specific kits for each analyte
2.5.
AC
(Gold analyzes Diagnostica, Belo Horizonte - Minas Gerais, Brazil).
Chemical analysis
For the chemical composition analysis of the diets and fillets, the methodology proposed by the [1] was used in which the moisture content was determined by drying the sample in an air circulation oven forced to 105°C until constant weight was reached (Method number 950.46), The ashes were determined by calcination of samples in a muffle furnace (Model 2000B, Belo Horizonte - Minas Gerais, Brazil) at 600°C method number 920.153, calcium and phosphorus were determined by
ACCEPTED MANUSCRIPT method number 968.08D, crude fiber method number 962.09 and protein by the Kjeldahl method (Model MA-036, Piracicaba - São Paulo, Brazil) method number 981.10. The total lipids were determined following [7] methodology. All analyses were performed in triplicate. Fatty acids derivatization was performed following the methodology described by [21]. Fatty acid methyl esters (FAME) were separated by gas chromatography (TRACE Ultra Thermo Scientific (Thermo Scientific, USA) equipped with flame ionization detector (FID) and a fused
T
silica capillary column (100 m x 0.25 mm i.d., 0.25 µm cyanopropyl, CP-7420 select FAME).
IP
Retention times and peak areas were determined using Chrom-Quest software (Thermo Scientific,
Statistical analysis
US
2.6.
CR
USA), following equipment cookbook and [59].
A one-way analysis of variance (ANOVA) was carried out with each diet as treatments and each
AN
tank as experimental units, the data were submitted to the Shapiro-Wilk normality test, at 5% probability. When significant differences were observed, the means were compared using Tukey’s
M
test at a 5% significance level. Regression analyses were performed to estimate the relationship between the treatments and responses. When more than one of the models showed significant
ED
coefficients, the regressions with the best biological explanation were chosen. All data were
Results
CE
3.
PT
analyzed using the software package R [53].
The results for growth performance at the end of the experiment are summarized in Table 3. There
AC
was a quadratic effect ( P < 0.05 ) based on increasing soybean oil inclusion on the variables daily weight gain, weight gain, final weight, carcass yield, visceral fat index, and hepatosomatic index. There was no effect ( P > 0.05 ) of soybean oil inclusion on feed conversion, fillet yield, dressed out yield and survival. Figure 1: Zootechnic parameters of Soybean oil inclusion in Tilapia’s nutrition
The optimal levels of inclusion of soybean oil in diets for Nile tilapia were determined by the adjustments of the quadratic regression models applied to the performance variables. The
ACCEPTED MANUSCRIPT inclusion of 45.29 g kg−1 (Figure 1a) and 54.36 g kg−1 of soybean oil provided better daily weight gain and higher final tilapia weight as shown in Figures S1a and S1b. The best results for carcass yield (Figure 1b), visceral fat and hepatosomatic index were observed with the inclusion of 37.60 kg−1, 55.17 kg−1 and 30.41 kg−1 of soybean oil respectively, as we can see in Figure S3a. The cost of the diet was lowest with the inclusion of 36.90 kg −1 of soybean oil (Figure 1c).
T
Table 3: Performance of fish fed with diets with increasing levels of soybean oil. Soybean oil inclusion g kg-1
SEM
p-value
Effect
407.33
409.00
0.64
>0.05
-
± 8.07
± 8.11
± 8.14
3.99 ±
4.11 ±
4.36 ±
4.17 ±
0.06
< 1.00
Quadratic
0.06 d
0.08 c
0.08 b
0.08 a
0.08 b
1.74 ±
1.76 ±
1.76 ±
1.77 ±
1.76 ±
0.03
0.03
0.03
0.03
0.03
Weight gain
185.66
199.66
205.56
218.00
208.33
(g) 3
± 2.99 d
± 3.198 ± 3.29 b
± 3.49
± 3.33 b
15.0
30.0
45.0
404.66
404.00
405.66
± 8.05
± 8.04
Daily weight
3.71 ±
gain (g) 1 Feed
590.33
(g)
± 9.45 d
611.33
625.33
617.33
± 9.65
± 9.78
± 10.00
± 9.88
cd
bc
b
ab
1.84 ±
1.76 ±
1.74 ±
1.67 ±
1.78 ±
(103) (US$/g)
0.03 a
0.02 b
0.02 bc
0.02 c
0.03 ab
AC
Diet cost rate
Carcass yield (%) 5 Dressed out yield (%) 6 Fillet yield
0.01
>0.05
-
2.87
< 1.00
Quadratic
88.27 ± 89.55 ± 89.93 ± 90.09 ± 89.44 ± 0.19 c
0.20 b
0.20 ab
0.20a
0.17
0.17
0.16
3.15
< 1.00
Quadratic
x106
3,12
< 1.00
Quadratic
x106
0.18
< 1.00
Quadratic
x106
0.20 b
51.18 ± 52.51 ± 52.10 ± 49.46 ± 50.17 ± 0.17
x106
a
603.66
CE
Final weight
PT
c
ED
rate 2
4
x106
M
conversion
AN
Initial weight
US
0.00
CR
60.0
IP
Parameters
0.48
>0.05
-
0.20
>0.05
-
0.16
33.72 ± 34.42 ± 34.00 ± 33.66 ± 33.80 ±
ACCEPTED MANUSCRIPT 0.09
0.08
0.08
0.08
Visceral fat
4.82 ±
4.19 ±
4.11 ±
4.01 ±
3.84 ±
index (%) 8
0.19 a
0.17 b
0.17 b
0.16 b
0.15 b
Hepatosomatic
3.66 ±
2.86 ±
3.05 ±
3.03 ±
3.49 ±
index (%) 9
0.24 a
0.198 c
0.20 bc
0.20bc
0.23 ab
100
100
100
100
100
Survival (%) 10
[*] 1 Daily weight gain [weight gain/day];
0.10
0.0003
Quadratic
0.10
0.0042
Quadratic
-
-
-
T
0.08
2
feed conversion [dry diet feed (g)/wet weight gained 4
Diet cost rate [cost of feed (per
CR
(g)]; 3 weight gain [final body weight initial body weight];
IP
(%) 7
kg)/weight gained x 1000]; 5 carcass yield assessments [(body weight (g) visceral fat weight
7
head(g)))/(body weight (g))x100];
US
(g))/(body weight (g))x100]; 6 dressed out yield [(body weight (g) (visceral weight (g) fish fillet yield [(fillet weigh (g)/body weight (g)) x100];
weight (g) x 100/final body weight (g)];
10
9
hepatosomatic index [liver
AN
visceral fat [visceral fat weight (g) x 100/final body weight (g)];
8
survival [(final number of fish/initial number of fish) x
M
100].
ED
Equation
gain
+ 2.50
kg-1)
R2
0
1
1
0.8929
0.0114
0.0009
< 1.00
PT
Y = 3.69
Adjusted
46.30
SEE
RPE
p-value
0.0010
< 1.00
x106
CE
Daily weight
SO (g
x106
x102 0SO
AC
3.00
x104 SO
2
Weight gain
Y = 184.4 + 1.25SO
1.39 x102 SO 2
45.29
0.8812
0.5699
0.0450
0.0007
0.0516
< 1.00
x106
ACCEPTED MANUSCRIPT Final weight
Y = 590.5
54.36
0.8264
0.8570
0.0677
0.0011
0.0385
< 1.00
x106
+ 1.11SO
1.02 x102 SO 2
Diet cost rate Y = 18.5
36.90
0.8377
0.0137
37.60
0.9183
0.1060
0.01081
0.00173
x102 SO
+ 9.70
1.30
3.20 x102 SO + 3.00
2
55.17
CE
x104 SO
Y = 3.58
ic index
4.50
AC
Hepatosomat
x106
x106
0.7518
0.0916
0.0072
< 1.00
0.0073
0.0003
0.0159
0.00416
x106
PT
index
ED
2
Y = 4.75
< 1.00
M
x103 SO
Visceral fat
0.0084
AN
x102 SO
US
2
yield
< 1.00
CR
+ 1.13
Y = 88.3
0.0004
IP
x101 SO
Carcass
0.00163
T
8.34
(10 3 )
0.0003
30.41
0.6860
0.1151
0.0091
0.0002
x102 SO
+ 7.00
x104 SO 2
* SO = Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE = Relative prediction error
ACCEPTED MANUSCRIPT
Results of hematological and biochemical are summarized in Table 4. For hematological and biochemical blood variables there was an effect ( P < 0.05 ) of increasing soybean oil levels on plasmatic cholesterol and triglycerides. The diet inclusion of soybean oil had no effect ( P > 0.05 ) on glucose, plasma proteins, hemoglobin, and total red cell hematocrit percentage for the blood
IP
T
parameters.
soybean oil. Parameters
CR
Table 4: Haematology and biochemical parameters of fish fed with diets with increasing levels of
Soybean oil inclusion g kg-1 15.0
30.0
45.0
56.04 ±
58.85 ±
58.07 ±
54.65 ±
55.69 ±
8.43
8.85
8.73
8.21
8.37
4.10 ±
4.31 ±
4.22 ±
0.17
0.17
0.17
9.18 ±
8.26 ±
0.84
Erythrocyte
3.97 ±
0.16
0.16
7.56 ±
8.68 ±
9.45 ±
0.75
0.69
0.80
0.88
2.20 ±
2.19 ±
2.17 ±
2.15 ±
2.33 ±
(106 mL-1)
0.10
0.10
0.10
0.10
0.10
Haematocrit
39.22 ±
39.11 ±
39.00 ±
39.11 ±
40.44 ±
0.75
0.74
0.74
0.74
0.77
Total
114.56
122.15
152.92
135.37
222.61
cholesterol
± 5.49 d
± 5.87
± 7.37 b
± 6.49 c
± 10.69
dL-1) Haemoglobin (g dL-1)
AC
(mg dL-1)
CE
(%)
ED
Protein (g
M
4.08 ±
PT
dL-1)
AN
Glucose (mg
60.0
US
0.00
cd
SEM
p-value
Effect
2.84
>0.05
-
0.05
>0.05
-
0.25
>0.05
-
0.03
>0.05
-
0.22
>0.05
-
2.38
< 1.00
Quadratic
x106
a
Triglycerides
184.04
205.94
220.37
268.33
269.35
(mg dL-1)
± 33.84
± 37.06
± 39.67
± 2.30
± 48.48
b
ab
ab
a
a
11.02
0.0143
-
Equation SO (mg−1
Adjusted
SEE
RPE
p-value
ACCEPTED MANUSCRIPT
Total
Y=
g)
R2
0
1
2
8.30
0.7533
3.3396
0.2637
0.0042
0.012303
0.000367
cholesterol 119.5002
0.5844SO + 0.03522SO2
T
* Means with different letter in the same raw are significantly different (Tukey) P < 0.05 ; SO =
IP
Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE =
CR
Relative prediction error
US
According to the quadratic regression analysis (Table4 and Figure S2b), the lowest plasma cholesterol level was observed with the addition of 8.30 g kg−1 of soybean oil in the diet. For
AN
triglycerides, statistical differences are shown ( P < 0.05 ) between the diets with the lowest level being observed in the lowest inclusion level and the highest level of triglycerides being seen with the inclusion of 60 g kg−1 of soybean oil.
M
For the chemical composition of the tilapia fillets (Table 5), a difference ( P < 0.05 ) was
ED
observed between treatments for crude protein and crude fat. There was no effect ( P > 0.05) on
PT
moisture and ash.
Table 5: Composition of tilapia fillets submitted to diets with increasing levels of soybean oil.
CE
Parameters (g kg-1)
0.00
15.0
30.0
45.0
60.0
SEM
p-value
766.0 ±
766.2 ±
766.1 ±
760.7 ±
765.3 ±
0.14
> 0.05
6.05
6.05
6.05
6.07
6.05
180.5 ±
173.9 ±
171.6 ±
171.5 ±
172.3 ±
0.11
0.0085
3.30 a
3.18 ab
3.14 b
3.14 b
3.15 b
38.3 ±
40.6 ±
43.0 ±
45.0 ±
43.3 ±
0.12
< 1.00
3.85 b
4.08 ab
4.33 a
4.53 a
4.35 a
10.9 ±
10.7 ±
11.2 ±
10.7 ±
11.1 ±
0.42
0.42
0.43
0.41
0.43
AC
Moisture
Crude protein
Crude fat
Ash
Soybean oil inclusion g kg-1
x106 0.01
> 0.05
*Means with different letter in the same raw are significantly different (Tukey) P < 0.05 ; SEM =
ACCEPTED MANUSCRIPT standard error of the mean;
The highest crude protein content in fillets was observed in the treatment without oil inclusion. For crude fat, there was a lower value for the control treatment and the 15.0 kg −1 of soybean oil inclusion. However, the other treatments had higher crude fat values. Fatty acid compositions of the tilapia fillets are summarized in Table 6. A total of 22 fatty
T
acids were determined in the tilapia fillets. Among saturated fatty acids (SFA), the highest
IP
concentration was observed for 16:0 (238.40 to 262.19 mg g −1); monounsaturated fatty acids
CR
(MUFA) was 18:1n-9 (215.36 to 277.61) and polyunsaturated fatty acids (PUFA), was 18:2n-6 (LA) (157.84 to 224.49).
US
There was a quadratic effect ( P < 0.05) , (Table6 and Figures 2a and 2b) when dietary soybean oil level increased for 16:1n-7; 18:3n-6, 18:3n-3 (LNA); 20:2n-6 and the total of omega 3
AN
( n-3) fatty acids.
M
Table 6: Fatty acid composition (mg g−1) of total lipids of fillet from Oreochromis niloticus after being fed diets with different soybean oil inclusions. Soybean oil inclusion g kg 1
14:0
0.00
15.0
30.0
45.0
60.0
SEM
p-value
Effect
29.57 ±
31.17 ±
30.34 ±
29.64 ±
35.01±
0.085
>0.05
-
3.00
2.92
2.85
3.37
3.73 ±
3.78 ±
4.07 ±
3.85±
0.054
2.56
-
0.06 c
0.06 b
0.07 c
2.85 4.25 ±
CE
15:0
0.06 a
17:0
18:0
16:1n-7
262.19 ±
AC
16:0
PT
(mg g−1)
ED
Fatty acid
0.05 c
238.40 ± 241.70 ± 260.19 ± 246.30 ±
38.80 a
35.28 b
35.77 b
38.51 a
36.45 b
2.39 ±
3.12 ±
6.68 ±
5.85 ±
3.82 ±
0.15 d
0.19 c
0.41 a
0.36 b
0.24 c
66.30 ±
65.99 ±
54.76 ±
47.14 ±
66.60 ±
6.62 a
4.37 a
2.39 b
1.13 c
9.99 a
68.21±
68.71 ±
62.26 ±
33.67 ±
22.25 ±
9.55 a
9.62 a
8.72 a
4.71 b
3.11 b
x1006 0.271
2.93
-
x1005 0.045
< 1.00
-
x106 0.212
< 1.00
-
x106 0.542
4.14
x105
Quadratic
ACCEPTED MANUSCRIPT 25.03 ±
1.17 a
0.83 bc
0.81 c
0.80 c
0.91 b
20:3n-6
20:4n-6 (AA)
2.46 c
2.35 d
6.88 ±
5.90 ±
5.61 ±
5.60 ±
5.33 ±
0.07 a
0.06 b
0.06 c
0.06 c
0.06 d
20.31 ±
17.45 ±
16.56 ±
16.53 ±
15.76 ±
0.22 a
0.19 b
0.18 c
0.18 c
0.17 d
< 1.00
0.014
< 1.00
0.042
2.36 a
4.61 ±
5.38 ±
6.49 ±
7.28 ±
5.61 ±
0.29 c
0.33 c
0.59 a
0.45 b
0.35 c
x106
10.24 ±
10.83 ±
12.05 ±
12.14 ±
12.08 ±
0.19 c
0.19 b
0.22 a
0.22 a
0.22 a
3.55 ±
3.92 ±
3.72 ±
3.71 ±
3.54 ±
0.09 c
0.04 a
0.04 b
0.04 b
0.04 c
6.57 ±
6.37 ±
8.08 ±
12.28 ±
16.13 ±
0.40 cd
0.39 d
0.49 c
0.75 b
0.99 a
38.06 ±
40.83 ±
43.24 ±
48.85 ±
49.28 ±
0.43 c
0.45 b
0.51 a
0.52 a
CE
27.83 ±
28.23 ±
27.73 ±
(LNA)
0.62 c
0.91 b
1.07 a
1.09 a
1.07 a
20:4n-3
2.18 ±
2.41 ±
2.32 ±
2.34 ±
2.21 ±
0.05 bc
0.06 a
0.06 abc
0.06 ab
0.06 bc
1.47 ±
1.34 ±
1.62 ±
2.37 ±
2.61 ±
(EPA)
0.11 b
0.10 b
0.13 b
0.18 a
0.20 a
22:6n-3
4.92 ±
5.41 ±
5.48 ±
5.48 ±
5.47 ±
(DHA)
0.08 b
0.09 a
0.09 a
0.09 a
0.09 a
n-6
200.87 ±
AC
23.70 ±
1.89 d
253.33 ± 273.56 ± 306.78 ± 311.12 ± 2.38 c
2.57 b
2.88 a
2.92 a
0.047
0.021
-
x106
2.34 a
16.19 ±
0.844
< 1.00
2.06 b
18:3n-3
-
x106
1.95 c
186.00 ± 196.99 ± 222.52 ± 224.49 ±
-
x106
1.66 d
0.40 d
20:5n-3
0.583
< 1.00
157.84 ±
-
x106
CR
20:2n-6
2.47 c
US
18:3n-6
2.60 b
< 1.00
AN
18:2n-6 (LA)
3.03 a
M
20:1n-9
238.46 ± 226.38 ± 225.94 ± 215.36 ±
0.103
T
21.89 ±
IP
22.23 ±
277.61 ±
18:1n-11
22:5n-6
22.62 ±
ED
18:1n-9
32.04 ±
PT
18:1n-7
-
x106 < 1.00
< 1.00
Quadratic
-
x106 0.038
2.92
-
x1006 0.102
< 1.00
-
x106 0.118
< 1.00
-
x106 0.124
< 1.00
Quadratic
x106 0.026
3.13
Quadratic
x103 0.014
2.15
Quadratic
x103 0.006
4.60
-
x1005 1.078
< 1.00
x106
-
ACCEPTED MANUSCRIPT
SFA
32.85 ±
37.26 ±
38.43 ±
38.02 ±
0.72 c
0.95 b
1.08 a
1.11 a
1.10 a
8.11 ±
7.72 ±
7.34 ±
7.98 ±
8.19 ±
0.21 a
0.20 ab
0.19 b
0.21 a
0.21 a
364.70 ±
342.41 ± 337.25 ± 346.88 ± 355.59 ±
3.90 a
MUFA
3.66 c
405.04 ±
3.71 bc
7.89 b
0.094
7.20 c
6.72 c
2.97
0.273
3.123e
1.138
-
3.26 b
3.62 a
3.66 a
PUFA/
0.62 ±
0.84 ±
0.92 ±
0.99 ±
0.98 ±
SFA
0.01 d
0.01 c
0.01 b
0.01 a
0.01 a
x106
0.037
-
x106
3.00 c
US
1.212
< 1.00
2.37 d
286.18 ± 310.82 ± 345.21 ± 349.14 ±
-
x1005
< 1.00
225.63 ±
-
x103
CR
8.37 b
< 1.00
x106
3.80 ab
353.15 ± 333.04 ± 303.63 ± 283.73 ±
9.60 a
PUFA
3.61 c
0.139
T
n-6/ n-3
24.76 ±
IP
n-3
-
x106 < 1.00
-
AN
*Results expressed as mean standard deviation of three replicates; values with different letters on the same line are significantly different (Tukey) P < 0.05 ; LA: linoleic acid; AA: arachidonic
M
acid; LNA: -linolenic acid; EPA: eicosapentaenoic acid; DHA: docosahexaenoic acid; SFA: total saturated fatty acids; MUFA: total of monounsaturated fatty acids; PUFA: total
ED
polyunsaturated fatty acids; n-6: total of omega 6 fatty acids; n-3: total omega-3 fatty acids;
saturated fatty acids. Equation
Adjusted
g−1)
R2
0
1
2
1.02
0.946
4.15
3.28x101
33.22
0.960
2.18x101 1.72 x102
CE
Y = 69.8
SO (g
AC
16:1n-7
PT
n-6/ n-3: rate of omega 6/omega 3; PUFA/SFA: total polyunsaturated fatty acids/total
SEE
RPE
p-value
5.20 x103
0.173
0.019
3.00 x104
0.024
< 1.00
2.97 x102 SO 1.46 x102 SO 2 18:3n-6
Y = 4.09
2.39 x101
x106
ACCEPTED MANUSCRIPT SO
3.60 x103 SO 2 18:3n-3
Y = 16.4
(LNA)
44.50
0.965
5.19 x101 4.10 x102
6.00 x104
0.016
< 1.00
x106
T
5.61x101 SO SO 2 Y = 2.21
29.25
0.780
3.12 x102 2.50 x103
1.13
0.923
7.95x102 6.30 x103
2.00 x104
5.30 x104
1.00 x104
0.018
3.35x103
M
(EPA)
AN
SO 2 Y = 1.39
0.043
US
1.17 x104 SO
20:5n-3
4.00 x105
CR
20:4n-3
IP
6.30 x103
ED
9.00 x104 SO
4.00 x104
PT
SO 2
*.SO = Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE
CE
= Relative prediction error
4.
AC
Figure 2: Fatty acid parameters of fillets of Tilapia fed with Soybean oil inclusion
Discussion
For fish, as well as tilapia, lipids perform essential functions in growth, feed efficiency, healthiness, kidney and gill function, neural and visual development, reproduction, and fillet quality [38]. Vegetable oils are easily found on the market and are important sources of lipids and energy for tropical fish and can be used depending on cost and availability [58]. No fish mortality was observed during the experimental period. [8] conducted a study evaluating different levels of energy and soybean oil inclusion in diets for tilapia also found no
ACCEPTED MANUSCRIPT effect of diets on the animal’s survival. The animals in the current study showed no mortality because they were well nourished and were raised in an adequate environment for their normal development. The quadratic effect observed for daily weight gain demonstrated an optimal level of soybean oil inclusion in the diet at 46.30 g kg−1, corresponding to a weight gain of 4.20 g day−1 and a total weight gain of 210 g. The lowest diet cost rate was observed to be at the inclusion of 36.90
T
g kg−1. Weight gain showed a quadratic effect with optimal growth occurring with the addition of
IP
54.36 kg−1 g of soybean oil in the diet.
CR
[55] found that the inclusion of oil above the diet needs of the animals caused a reduction in the rate of enzymatic reactions and specific activity of the catalytic efficiency of glucose
US
6-phosphate dehydrogenase in the liver of trout. This may explain the decline in growth with the excessive oil level in the diet observed in the current paper.
AN
The feed conversion rate of diets with the high content of soybean oil was similar between treatments, showing that the energy level of the different diet did not influence the feed utilization
M
of the animals. [42] observed feed conversion values of 1.69 for fingerlings Nile tilapia in diets supplemented with 3 and 4.8% of fat. In the present study, the feed conversion rate was superior
ED
due to the animal breeding phase being different. It is known that feed conversion increases with the increasing weight of the animal, namely in the early stages, so it is natural that the feed
PT
conversion is higher in the final stages of farming. The diets evaluated in the current study were in a relationship between digestible protein
CE
and digestible energy between 18.27 and 20.28 mg DP kJ−1, and the best ratio of digestible protein to digestible energy was observed to be 18.77 mg DP kJ−1 with the inclusion of around 45.0 g kg 1
AC
of soybean oil addition. While [10] found an ideal ratio of digestible protein and digestible energy to be 20.45 mg DP kJ−1 for tilapia larvae in the sexual reversion phase. It is essential to consider the protein to energy ratio for a good development of fish, taking into account the impacts of the management and environment [9]. The utilization of an adequate level of oil in feeds for fish brings an opportunity for lower production cost as shown in the diet cost rate in the present study. The highest carcass yield was observed for the inclusion of 37.60 g kg −1 of soybean oil in the diet when using quadratic regression analysis (Table 3). Even though tilapia is a species that is good at utilizing carbohydrates, apparently there was an excess of starch, causing an increase in fat in the abdominal cavity. The diet without soybean oil inclusion provided an increase in the visceral
ACCEPTED MANUSCRIPT fat deposition; this diet contained the highest starch content. Starch may have influenced visceral fat, as increased oil content and decreased starch in the diet resulted in lower fish visceral fat. The higher proportion of carbohydrates may explain increased visceral fat deposition. According to the [48], the body fat deposition content in fish can increase when the diet is formulated with starch above the capacity of utilization as an energy source. However, for this phase of development, with a fully functional digestive tract, there is insufficient information
T
about starch inclusion limit in diets for tilapia.
IP
When it comes to other species, such as mammals, the mechanisms that are potentially
CR
responsible for the lower body fat deposition are the reduction of energy consumption by suppressing appetite and increasing energy expenditure in muscles by lipolysis [36]. Although the
US
fish mechanisms are not clearly understood, they are probably related to the inhibition of lipids absorption and even lipid oxidation [40].
AN
According to the quadratic regression equation for the hepatosomatic index, the lowest value was estimated to be at 30.41 g kg−1 of soybean oil inclusion. In a study by [37], evaluating
M
the interaction between crude fiber and soybean oil in diets for tilapia, the authors observed an increase in the hepatosomatic index, which was also observed by [3]. The hepatosomatic index is
ED
directly linked to glycogen accumulation in the liver and consequently causes an increase in liver weights [16, 45]. Therefore, in the present paper, the diet with the higher content of starch
PT
demonstrated a higher hepatosomatic index, results similar observed by [6] in their study with hybrid catfish, using similar content of starch in its diets. This fact, is most likely related to
CE
increased starch metabolism, through glycogenesis catalyzed by glycogen synthase [48] and glycogen deposition in the liver [28, 6, 64].
AC
The inclusion of soybean oil in the diet had no effect ( P > 0.05 ) on the glucose, total serum proteins, hemoglobin, erythrocyte, and hematocrit of the blood. Evaluating the interaction of the inclusion of 0 to 6% sunflower oil and linseed in diets for tilapia raised in reduced temperatures, [2] observed reductions in erythrocyte levels (2.02 to 1.76) in fish fed 4% of sunflower oil and 2% of flaxseed oil and 3.82 to 3.05 of total protein in fish fed 6% flaxseed oil. The authors concluded that diets with a low proportion of omega-6 and omega-3 fatty acids lead to a reduction in blood profile when the animals are submitted to a thermal challenge. According to the quadratic regression equation, the lowest plasma cholesterol level was observed with the addition of 8.30 kg−1 g of soybean oil inclusion, indicating an upward trend for
ACCEPTED MANUSCRIPT cholesterol as soybean oil inclusion is increased in the diet. This fact is possibly correlated with the characteristic of a body’s cholesterol retention and cholesterol being a precursor of bile salts necessary for the digestion of ingested lipids [46]. The total plasma cholesterol content may be regarded as indicative of the health of fish, when tilapia are kept in unhealthy environments reductions in plasma cholesterol levels have been observed [63]. [4] found a mean value of 120 mg dL−1 for plasma cholesterol, evaluating the
T
interaction of soybean oil and palm oil, as a lipid source, when feeding palm meal to tilapia. [52]
IP
observed that the use of vegetable oils in diets for Acanthopagrus schlegeli contributes to lowered
CR
plasma cholesterol. According to [62], excessive intake of feeds rich in fat content and cholesterol may increase its concentration in the blood. However, it is noteworthy that this molecule is a
US
precursor of hormones such as sterol hormones, which are responsible for the growth of fish, and vitamins needed for vital processes of reproduction and maintenance of health. Nevertheless, there
AN
is no consensus on cholesterol levels, and comparisons are often made with different diets and rearing systems, mainly when energy sources are used in the diet [47]. However, in this study, the
M
total plasma cholesterol contents are in agreement with those proposed by [32] for tilapia in intensive systems.
ED
The triglyceride levels show statistical differences ( P < 0.05 ) between the diets used. Similar results were observed by [20] as they studied the effect of lipid levels on growth and body
PT
composition of juvenile grass carp. Their work showed a linear increase in triglyceride content as the lipid content was increased in the feed. In our study, it is important to note that the increase of
CE
total cholesterol and total triglycerides according to the increased inclusion of soybean oil does not adversely influence the visceral fat deposition since there was a reduction of fat deposited in the
AC
viscera with the increasing inclusion of oil. In the analysis of the chemical composition of the tilapia fillets, the crude protein content the fillets found in this study was similar to those reported by [5], which evaluated the effect of the inclusion of palm oil sources for Oreochromis sp., obtaining results from 16.9 to 17, 9%. Similar results were also observed by [25], who evaluated the influence of inclusion of different fat sources (soybean oil, corn oil, linseed oil, fish oil and olive oil) on muscle composition and concentration of plasma lipoproteins for O. niloticus, obtaining crude protein values from 16.28 to 18.27%. The values vary according to the inclusion of oils. Animals fed with diets isonitrogenous, isocaloric, and with no changes in lipid levels
ACCEPTED MANUSCRIPT usually do not differ in the basic chemical composition [41]. However, [26] stated that the variation in chemical composition of fish might occur due to endogenous and exogenous factors such as genetics, size, sex, reproductive stage, environmental factors, temperature and season, in addition to nutrient intake. In addition, there may be a higher concentration of fat/protein in different parts of the body, in order to eliminate this factor, the fish fillets were ground and homogenized for chemical analysis in the present study.
T
Lipid nutrition is directly related to the quality of fish fillet [34], which influences the
IP
commercial value of the fish. Generally, in older fish, there is greater fat deposition compared to
CR
young fish [49], this is due to the younger fish having a higher growth rate in which dietary energy and protein is used for growth rather than storage for energy reserves [15]. Evaluating the effect of
US
soybean oil to fish oil ratio in Acanthopagrus schlegeli diets, [52] observed differences in the lipid composition of the fillets and found that soybean oil positively influences the deposition of
AN
intramuscular fat as well as decreases intraperitoneal fat deposition. Production of high-quality meat products has been a big focus lately [18]. Optimal finisher
M
diet formulations not only provide good feed efficiency and growth rate, they reduce costs, and also affect the carcass and fillet quality. The composition of the diets in this study had a direct
ED
effect (Table 6 and Figure 2a and 2b) on fillet composition and fatty acid composition. Nile tilapia metabolism requires n-6 fatty acids in their diets to maximize development. They also tend to store
PT
large amounts of these compounds in their musculature [29, 25], this propriety can be used to produce healthier products for human consumption [61].
CE
The increase of long-chain polyunsaturated essential fatty acid levels, (20:5-n3 (EPA) and (22:6-n3 (DHA)), may be attributed to the concentration -linolenic acid in, a precursor of the
AC
n-3 fatty acid series, in the diets. [12], studied the inclusion of -linolenic acid in the nutrition of Nile tilapia. They attribute the increase of DHA to the bioconversion of -linolenic acid, as a substrate, of the -6/-5 desaturases and elongase enzymes in the metabolism of Nile tilapia. Also, [44] attributed the increase of PUFAs in carcasses to the antioxidant activity of vitamin E concentrations incorporated in diets. The diets evaluated in our current work have higher levels of soybean oil that contains a great quantity of vitamin E and PUFAs [13], this can provide greater nutritional values to the fillets. All fillets in this study showed higher PFUA to SFA ratios than the recommended minimum by the Department of Health and Social Security [17].
ACCEPTED MANUSCRIPT 5.
Conclusion
It is concluded that diets with soybean oil inclusion around 45.0 g kg −1, provide better growth performance of tilapia during the finishing phase. Dietary supplementation of soybean oil was effective at improving nutrition values of the meat of tilapia as well as achieving above the recommended PUFA to SFA ratio values described by the Department of Health and Social
References
CR
6.
IP
T
Security.
References
AOAC (2005). Association of Official Analytical Chemists (AOAC). (19th ed.). Hoboken,
US
[1]
NJ, USA: John Wiley & Sons, Inc. doi:10.1002/0471740039.vec0284. Araujo, D., Pezzato, A. C., Barros, M. M., Pezzato, L. E., & Nakagome, F. K. (2011).
AN
[2]
Hematologia de tilápias-do-nilo alimentadas com dietas com óleos vegetais e estimuladas
M
pelo frio. Pesquisa Agropecuária Brasileira, 46, 294–302. doi:10.1590/s0100-204x2011000300010. Atwood, H. L., Tomasso, J. R., Webb, K., & Gatlin, D. M. (2003). Low-temperature
ED
[3]
tolerance of nile tilapia, oreochromis niloticus: effects of environmental and dietary
PT
factors. Aquaculture Research, 34, 241–251. doi:10.1046/j.1365-2109.2003.00811.x. de Azevedo, R. V., Tonini, W. C. T., & Braga, L. G. T. (2013). Óleo e torta de dend^e em
CE
[4]
rações para juvenis de tilápia-do-nilo. Pesquisa Agropecuária Brasileira, 48, 1028–1034.
[5]
AC
doi:10.1590/s0100-204x2013000800031. Bahurmiz, O. M., & Ng, W.-K. (2007). Effects of dietary palm oil source on growth, tissue fatty acid composition and nutrient digestibility of red hybrid tilapia, oreochromis sp., raised from stocking to marketable size. Aquaculture, 262, 382–392. doi:10.1016/j.aquaculture.2006.11.023. [6]
Bernardes, C.,, Navarro, R., Santos, B. G., Silva, R. F.,, & and (2016). Effects of dietary carbohydrate/lipid ratios on growth, body composition, and nutrient utilization of hybrid catfish (pseudoplatystoma reticulatum x leiarius marmoratus). Revista Colombiana de Ciencias Pecuarias, 29. URL:
ACCEPTED MANUSCRIPT https://doi.org/10.17533%2Fudea.rccp.v29n1a08. doi:10.17533/udea.rccp.v29n1a08. [7]
Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Canadian journal of biochemistry and physiology, 37, 911–917.
[8]
Boscolo, W. R., Hayashi, C., & Meurer, F. (2002). Digestibilidade aparente da energia e
[9]
IP
niloticus, l.). Revista Brasileira de Zootecnia, 31, 539–545.
T
nutrientes de alimentos convencionais e alternativos para a tilápia do nilo (oreochromis
Boscolo, W. R., Hayashi, C., & Meurer, F. (2002). Farinha de varredura de mandioca
l.). Revista Brasileira de Zootecnia, 31, 546–551.
Boscolo, W. R., Hayashi, C., Meurer, F., Feiden, A., & Wolff, L. (2004). Desempenho e
US
[10]
CR
(manihot esculenta) na alimentação de alevinos de tilápia do nilo (oreochromis niloticus
características de carcaça de tilápias do nilo (oreochromis niloticus l.) alimentadas com
AN
rações contendo diferentes níveis de gordura. Acta Scientiarum. Animal Sciences, 26, 443–447.
Boscolo, W. R., Signor, A., Feiden, A., Bombardelli, R. A., Signor, A. A., & Reidel, A.
M
[11]
(2005). Energia digestível para larvas de tilápia-do-nilo (oreochromis niloticus) na fase de
ED
reversão sexual. Revista Brasileira de Zootecnia, 34, 347 1813–1818. doi:10.1590/s1516-35982005000600003. Chen, C., Guan, W., Xie, Q., Chen, G., He, X., Zhang, H., Guo, W., Chen, F., Tan, Y., &
PT
[12]
Pan, Q. (2018). n-3 essential fatty acids in nile tilapia, oreochromis niloticus :
CE
Bioconverting LNA to DHA is relatively efficient and the LC-PUFA biosynthetic pathway is substrate limited in juvenile fish. Aquaculture, 495, 513–522.
[13]
AC
doi:10.1016/j.aquaculture.2018.06.023. Clemente, T. E., & Cahoon, E. B. (2009). Soybean oil: Genetic approaches for modification of functionality and total content. PLANT PHYSIOLOGY, 151, 1030–1040. doi:10.1104/pp.109.146282. [14]
Collier, H. B. (1944). Standardization of blood haemoglobin determinations. Canadian Medical Association Journal, 50, 550.
[15]
Contreras-Guzmán, E. S. (1994). Bioquímica de pescados e derivados. FUNESP.
[16]
Cyrino, J. E. P., Portz, L., & Martino, R. C. (2000). Retenção de proteína e energia em juvenis de. Scientia Agricola, 57, 609–616.
ACCEPTED MANUSCRIPT [17]
DHSS, U. (1994). Nutritional Aspects of Cardiovascular Disease. Technical Report Report on Health and Social Subjects London.
[18]
Dokmanovic, M., Baltic, M. Z., Duric, J., Ivanovic, J., Popovic, L., Todorovic, M., Markovic, R., & Pantic, S. (2015). Correlations among stress parameters, meat and carcass quality parameters in pigs. Asian-Australasian Journal of Animal Sciences, 28, 435–441. doi:10.5713/ajas.14.0322.
T
Drotningsvik, A., Mjøs, S. A., Pampanin, D. M., Slizyte, R., Carvajal, A., Remman, T.,
IP
[19]
Høgøy, I., & Gudbrandsen, O. A. (2016). Dietary fish protein hydrolysates containing
CR
bioactive motifs affect serum and adipose tissue fatty acid compositions, serum lipids, postprandial glucose regulation and growth in obese zucker fa/fa rats. British Journal of
[20]
US
Nutrition, 116, 1336–1345.
Du, Z.-Y., Liu, Y.-J., Tian, L.-X., Wang, J.-T., Wang, Y., & 373 Liang, G.-Y. (2005).
AN
Effect of dietary lipid level on growth, feed utilization and body composition by juvenile grass carp (ctenopharyngodon idella). Aquaculture Nutrition, 11, 139–146. Duckett, S. K., Andrae, J. G., & Owens, F. N. (2002). Effect of high-oil corn or added corn
M
[21]
oil on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in
ED
beef steers fed finishing diets. Journal of Animal Science, 80, 3353–3360. doi:10.2527/2002.80123353x. FAO (2014). The State of World Fisheries and Aquaculture 2014. Rome.
[23]
FAO (2016). The state of world fisheries and aquaculture 2016. Technical Report FAO
PT
[22]
[24]
CE
Rome. URL: http://www.fao.org/3/a-i5555e.pdf. FAO (2018). The state of world fisheries and aquaculture 2018: meeting the sustainable
[25]
AC
development goals. Rome: UN. Ferreira, M. W., de Araujo, F. G., Costa, D. V., Rosa, P. V., Figueiredo, H. C., & Murgas, L. D. (2011). Influence of dietary oil sources on muscle composition and plasma lipoprotein concentrations in nile tilapia, oreochromis niloticus. Journal of the World Aquaculture Society, 42, 24–33. [26]
Fonseca-Madrigal, J., Karalazos, V., Campbell, P., Bell, J. G., & Tocher, D. R. (2005). Inuence of dietary palm oil on growth, tissue fatty acid compositions, and fatty acid metabolism in liver and intestine in rainbow trout (oncorhynchus mykiss). Aquaculture Nutrition, 11, 241–250.
ACCEPTED MANUSCRIPT [27]
Goldenfarb, P. B., Bowyer, F. P., Hall, E., & Brosious, E. (1971). Reproducibility in the hematology laboratory: the microhematocrit determination. American Journal of Clinical Pathology, 56, 35–39.
[28]
Hemre, G.-I., Monnsen, T., & Krogdahl, A. (2002). Carbohydrates in fish nutrition: effects on growth, glucose metabolism and hepatic enzymes. Aquaculture Nutrition, 8, 175–194. URL: https://doi.org/10.1046%2Fj.1365-2095.2002.00200.x.
IP
[29]
T
doi:10.1046/j.1365-2095.2002.00200.x.
Higuchi, L. H., Feiden, A., Matsushita, M., Santarosa, M., Zanqui, A. B., Bittencourt, F., &
CR
Boscolo, W. R. (2013). Quantificação de ácidos graxos de alevinos de tilápia do nilo (oreochromis niloticus) alimentados com diferentes fontes de óleos vegetais. Semina:
[30]
US
Ciências Agrárias, (pp. 1913–1924).
Hildsorf, A. (1995). Genética e cultivo de tilápias vermelhas, uma revisão. Boletim do
[31]
AN
Instituto de Pesca, 22, 73–84.
Hooley, C. G., Barrows, F. T., Paterson, J., & Sealey, W. M. (2014). Examination of the
M
effects of dietary protein and lipid levels on growth and stress tolerance of juvenile tilapia,oreochromis niloticus. Journal of the World Aquaculture Society, 45, 115–126.
[32]
ED
doi:10.1111/jwas.12110.
Hrubec, T. C., Cardinale, J. L., & Smith, S. A. (2000). Hematology and plasma chemistry
PT
reference intervals for cultured tilapia (oreochromis hybrid). Veterinary clinical pathology, 29, 7–12.
Izquierdo, M., Obach, A., Arantzamendi, L., Montero, D., Robaina, L., & Rosenlund, G.
CE
[33]
(2003). Dietary lipid sources for seabream and seabass: growth performance, tissue
[34]
AC
composition and flesh quality. Aquaculture Nutrition, 9, 397–407. Justi, K., Hayashi, C., Visentainer, J., De Souza, N., & Matsushita, M. (2003). The influence of feed supply time on the fatty acid profile of nile tilapia (oreochromis niloticus) fed on a diet enriched with n-3 fatty acids. Food chemistry, 80, 489–493. [35]
Kaushik, S. (1990). Importance des lipides dans la alimentation des poissons. Aqua Revue, 29, 9–16.
[36]
Kennedy, A., Martinez, K., Schmidt, S., Mandrup, S., LaPoint, K., & McIntosh, M. (2010). Antiobesity mechanisms of action of conjugated linoleic acid. The Journal of nutritional biochemistry, 21, 171–179.
ACCEPTED MANUSCRIPT [37]
Lanna, E. A. T., Pezzato, L. E., Furuya, W. M., Vicentini, C. A., Cecon, P. R., & Barros, M. M. (2004). Fibra bruta e óleo em dietas práticas para alevinos de tilápia do nilo (oreochromis niloticus). Revista Brasileira de Zootecnia, (pp. 2177–2185).
[38]
Lim, C., & Webster, C. D. (2001). Nutrition and fish health. Food Products Press New York.
[39]
Luna, S., & Teixeira, M. (2007). Eutanásia: considerações éticas e indicações técnicas.
Makol, A., Torrecillas, S., Fernández-Vaquero, A., Robaina, L., Montero, D., Caballero,
IP
[40]
T
Revista CFMV, 13, 60–69.
CR
M. J., Tort, L., & Izquierdo, M. (2009). Effect of conjugated linoleic acid on dietary lipids utilization, liver morphology and selected immune parameters in sea bass juveniles
and Molecular Biology, 154, 179–187.
Menoyo, D., Lopez-Bote, C. J., Bautista, J. M., & Obach, A. (2003). Growth, digestibility
AN
[41]
US
(dicentrarchus labrax). Comparative Biochemistry and Physiology Part B: Biochemistry
and fatty acid utilization in large atlantic salmon (salmo salar) fed varying levels of n-3 and
[42]
M
saturated fatty acids. Aquaculture, 225, 295–307. Meurer, F., Hayashi, C., Boscolo, W. R., & Soares, C. M. (2002). Lipídeos na alimentação
Zootecnia, 31, 566–573.
Moreira, P. O., Rocha, J. D. M., da Silva, T. C., Bittencourt, F., Dacley Hertes, N., &
PT
[43]
ED
de alevinos revertidos de tilápia do nilo (oreochromis niloticus l.). Revista Brasileira de
Boscolo, W. R. (2015). Óleo de soja utilizado pré e pós-processamento de rações para
[44]
CE
tilápia do nilo. Bol. Inst. Pesca, São Paulo, 41, 547–555. Navarro, R. D., Navarro, F. K. S. P., Filho, O. P. R., Ferreira, W. M., Pereira, M. M., &
AC
Filho, J. T. S. (2012). Quality of polyunsaturated fatty acids in nile tilapias (oreochromis niloticus) fed with vitamin e supplementation. Food Chemistry, 134, 215–218. doi:10.1016/j.foodchem.2012.02.097. [45]
Navarro, R. D., Pinto, O., Filho, R., & Calado, L. L. (2006). Comparação morfometrica e índices somáticos de machos e fêmeas do lambari prata ( Astayanax scabripinnis Jerenyns, 1842) em diferente sistema de cultivo. Zootecnia Tropical, 24, 165–176. doi:S0798-72692006000200006.
[46]
Nelson, D. L., Lehninger, A. L., & Cox, M. M. (2008). Lehninger principles of biochemistry. Macmillan.
ACCEPTED MANUSCRIPT [47]
Neu, D., Furuya, W., Boscolo, W., Potrich, F., Lui, T., & Feiden, A. (2012). Glycerol inclusion in the diet of nile tilapia (oreochromis niloticus) juveniles. Aquaculture Nutrition, 19, 211––217. doi:10.1111/j.1365-2095.2012.00968.x.
[48]
NRC (2011). Nutrient Requirements of Fish and Shrimp. Washington, DC: The National Academies Press.
[49]
Ogawa, M., & Maia, E. L. (1999). Manual da Pesca Ciência e tecnologia do pescado. São
IP
[50]
T
Paulo: Varela.
Okamura, D., de Araújo, F. G., Rosa, P. V. e., de Freitas, R. T. F., Murgas, L. D. S., &
CR
Cesar, M. P. (2010). Influência da concentração de benzocaína e do comprimento dos peixes na anestesia e na recuperação de tilápias-do-nilo. Revista Brasileira de Zootecnia,
[51]
US
39, 971–976. doi:10.1590/S1516-35982010000500005. Omasaki, S., Janssen, K., Besson, M., & Komen, H. (2017). Economic values of growth Aquaculture, 481, 124–132. URL:
AN
rate, feed intake, feed conversion ratio, mortality and uniformity for nile tilapia.
M
https://doi.org/10.1016%2Fj.aquaculture.2017.04.013. doi:10.1016/j.aquaculture.2017.04.013. Peng, S., Chen, L., Qin, J. G., Hou, J., Yu, N., Long, Z., Ye, J., & Sun, X. (2008). Effects of
ED
[52]
replacement of dietary fish oil by soybean oil on growth performance and liver
PT
biochemical composition in juvenile black seabream, Acanthopagrus schlegeli. Aquaculture, 276, 154–161. doi:10.1016/j.aquaculture.2008.01.035. R Development Core Team (2013). R: A language and environment for statistical
CE
[53]
computing. R Foundation for Statistical Computing, Vienna, Austria. URL
[54]
AC
http://www.R-project.org/. Ribeiro, P. A. P., Logato, P. V. R., Paula, D. A. d. J., Costa, A. C., Murgas, L. D. S., & de Freitas, R. T. F. (2008). Efeito do uso de óleo na dieta sobre a lipogênese e o perfil lipídico de tilápias-do-nilo. Revista Brasileira de Zootecnia, 37, 1331–1337. doi:10.1590/S1516-35982008000800001. [55]
Sánches-Muros, M. J., García-Rejón, L., Lupiáñez, J. A., & Higuera, M. (1996). Long-term nutritional effects on the primary liver and kidney metabolism in rainbow trout (Oncorhynchus mykiss). II. Adaptive response of glucose 6-phosphate dehydrogenase activity to high-carbohydrate/low-protein and high-fat/non-carbohydrate diets.
ACCEPTED MANUSCRIPT Aquaculture Nutrition, 2, 193–200. doi:10.1111/j.1365-2095.1996.tb00059.x. [56]
Sousa, Á. B. B. d., Júnior, S., de Oliveira, O., Visentainer, J. V., & Almeida, N. M. d. (2017). Total lipid nutritional quality of the adipose tissue from the orbital cavity in nile tilapia from continental aquaculture. Acta Scientiarum. Animal Sciences, 39, 335–341.
[57]
Steffens, W. (1997). Effects of variation in essential fatty acids in fish feeds on nutritive
T
value of freshwater fish for humans. Aquaculture, 151, 97–119.
Vargas, R. J., Souza, S. M., Tognon, F., Gomes, M. E., & Kessler, A. d. M. (2007).
CR
[58]
IP
doi:10.1016/S0044-8486(96)01493-7.
Desempenho de alevinos de tilápias do nilo alimentados com dietas contendo diferentes
[59]
US
fontes de lipídeos. Revista Brasileira de Agrociência, 13, 377–381. Visentainer, J. V. (2012). Aspectos analíticos da resposta do detector de ionização em
AN
chama para ésteres de ácidos graxos em biodiesel e alimentos. Química Nova, 35, 274–279. doi:10.1590/s0100-40422012000200008. Wang, J.-T., Liu, Y.-J., Tian, L.-X., Mai, K.-S., Du, Z.-Y., Wang, Y., & Yang, H.-J.
M
[60]
(2005). Effect of dietary lipid level on growth performance, lipid deposition, hepatic
ED
lipogenesis in juvenile cobia (rachycentron canadum). Aquaculture, 249, 439–447. doi:10.1016/j.aquaculture.2005.04.038. Wood, J., Richardson, R., Nute, G., Fisher, A., Campo, M., Kasapidou, E., Sheard, P., &
PT
[61]
Enser, M. (2004). Effects of fatty acids on meat quality: a review. Meat Science, 66, 21–32.
[62]
CE
doi:10.1016/s0309-1740(03)00022-6. Xu, W.-n., Li, J.-y., Li, X.-f., Huang, G.-q., Liu, W.-b. et al. (2013). Alterations of liver
AC
histology and blood biochemistry in blunt snout bream megalobrama amblycephala fed high-fat diets. Fisheries Science, 79, 661–671. [63]
Yoneyama, Y., Yonemori, Y., Murata, M., Ohnuki, H., Hibi, K., Hayashi, T., Ren, H., & Endo, H. (2009). Wireless biosensor system for real-time cholesterol monitoring in fish “Nile tilapia”. Talanta, 80, 909–15. doi:10.1016/j.talanta.2009.08.014.
[64]
Zhou, P., Wang, M., Xie, F., Deng, D.-F., & Zhou, Q. (2016). Effects of dietary carbohydrate to lipid ratios on growth performance, digestive enzyme and hepatic carbohydrate metabolic enzyme activities of large yellow croaker (larmichthys crocea). Aquaculture, 452, 45–51. URL:
ACCEPTED MANUSCRIPT https://doi.org/10.1016%2Fj.aquaculture.2015.10.010.
AC
CE
PT
ED
M
AN
US
CR
IP
T
doi:10.1016/j.aquaculture.2015.10.010.
ACCEPTED MANUSCRIPT
AC
CE
PT
ED
M
AN
US
CR
IP
T
Soybean oil in increasing levels affect the fishes muscle composition; Vegetable oil has influence in fish nutrition; Soybean oil as energy and fat acids source; Soybean oil does increasing tilapias' development.
Antonio Cesar Godoy, Oscar Oliveira Santos, Jarred Hugh Oxford, Iury Walysson de Amorim Melo, Rômulo Batista Rodrigues, Dacley Neu, Ricardo Nunes, Wilson Rogério Boscolo PII: DOI: Article Number: Reference:
S0044-8486(19)30467-3 https://doi.org/10.1016/j.aquaculture.2019.734324 734324 AQUA 734324
To appear in:
aquaculture
Received date: Revised date: Accepted date:
23 February 2019 15 June 2019 18 July 2019
Please cite this article as: A.C. Godoy, O.O. Santos, J.H. Oxford, et al., Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: Economic, zootechnical and nutritional meat improvements, aquaculture, https://doi.org/10.1016/ j.aquaculture.2019.734324
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Soybean oil for Nile tilapia (Oreochromis niloticus) in finishing diets: economic, zootechnical and nutritional meat improvements Antonio Cesar Godoya,*, Oscar Oliveira Santosa, Jarred Hugh Oxfordb, Iury Walysson de Amorim
Universidade Estadual de Maringá. Av. Colombo, 5790 - Centro de Ciências Exatas -
IP
a
T
Meloc, Rômulo Batista Rodriguesc, Dacley Neuc, Ricardo Nunesc, Wilson Rogério Boscolo c,*
b c
CR
Departamento de Química. Maringá/PR, Brazil
University of Georgia - Poultry Science Department. Athens GA. USA
Universidade Estadual do Oeste do Paraná. Centro de Engenharias e Ciências Exatas. Rua da
US
Faculdade, 645, Toledo/PR, Brazil.
AN
Abstract
The aim of this study was to evaluate different levels of soybean oil inclusion in diets for Nile
M
tilapia. 270 tilapia, with an initial weight of 425.33 ± 32.37 g and initial length of 25.53 ± 2.00 cm, were randomly distributed in 15 tanks. The diets were formulated to be isonitrogenous (30%)
ED
using five increasing levels of soybean oil (0.00; 15.00; 30.00; 45.00 and 60.00 g kg −1) with each level having three replicates. The evaluated parameters were: performance (survival, daily weight
PT
gain, feed conversion, hepatosomatic index, visceral fat, carcass yield, fillet yield and dressed out yield), hematological and biochemical blood aspects, total protein, cholesterol, glucose,
CE
triglycerides, hemoglobin, hematocrit and red blood cells, and the chemical composition of the fillets (moisture, crude protein, lipids, and ash). Quadratic effects ( P < 0.05 ) were observed for
AC
the variables: daily weight, weight gain, final weight, carcass yield, visceral fat, and hepatosomatic index. For blood variables, a quadratic effect ( P < 0.05 ) for total cholesterol was observed. For chemical composition analyses of the fillets, there were no effects ( P > 0.05 ) observed with the increase of soybean oil. A total of 22 fatty acids were determined in the fillets of the tilapia fed with different soybean oil levels. Among fatty acids, the highest concentrations were for 16:0 *
Corresponding author
address:
[email protected]
[email protected] (Wilson Rogério Boscolo)
(Antonio
Cesar
Godoy),
ACCEPTED MANUSCRIPT (238.40 to 262.19 mg g−1); 18:1n-9 (215.36 to 277.61) and 18:2n-6 (LA) (157.84 to 224.49). It is concluded that the addition of around 45.0 g kg−1 of soybean oil to the finisher diet of Nile tilapia provides better growth performance. Animals fed the diet, used the nutrients to produce recommended PUFA/SFA values. Dietary supplementation was also effective at improving the nutrition values of the meat.
T
Keywords: economic improvement, fish growth performance, essentials fatty acid, human
CR
1.
IP
nutrition
Introduction
US
In aquaculture one of the most prominently farmed species is the Nile tilapia (Oreochromis niloticus), this can be attributed to the rapid growth, hardiness, large fillet yield, absence of
AN
intramuscular bones, and excellent acceptance by the consumer market [30]. The tropical fish most farmed in the world [51] and it still growing [22, 23, 24] it is estimated that the production of
M
tilapia will reach approximately 6.6 million tons by 2030 [51].
ED
Fish, in general, have excellent nutritional properties, with balanced protein compositions, essential fatty acids, vitamins, and minerals. Another benefit of fish is the ability to improve the nutritional quality of the meat through supplementation with different additions of fatty acids and
PT
vitamins sources in the diet [44, 19, 56].
There are many components in fish diets with the lipids being considered as an essential
CE
energy source that can significantly influence feed conversion and fish growth [57]. In intensive fish farming, high levels of fat sources in the diet are used in order to meet the fish’s energy
AC
requirements; this is done to prevent the breakdown of protein to be used as an energy source in the body [25]. However, excessive energy supply can influence the animal’s metabolism and cause adverse effects on growth, carcass composition with undesirable fat accumulation in the viscera and abdominal cavity. As well as reducing the percentage of fillet yield and the commercial value of the fish, it has been shown that the tissue lipid composition reflects the feed and can be changed by the manipulation of the diet [35, 60, 54]. Vegetable oils are regarded as valuable energy sources for tropical fish, because they are easily found on the market, and are used according to their availability and cost of the raw
ACCEPTED MANUSCRIPT materials in each region. Soybean oil is more frequently used due to its low cost when compared to other energy sources, and its overall composition meets the animal’s nutritional demands [33]. Soybean oil has an apparent digestibility of 89.85% and 8,485 kcal kg −1 of digestible energy, demonstrating that this nutrient is well used by the species O. niloticus [8]. Some studies have evaluated the inclusion of soybean oil in diets of Nile tilapia showing good results in the literature. The addition of 5.9% of soybean oil in the diet increases the carcass
T
and fillet yields of the tilapia [10, 43]. However, few studies have evaluated the inclusion of
IP
soybean oil in the finisher phase diets of tilapia.
CR
The inclusion of lipids in fish feed might be a potential mechanism to reduce protein levels and commercial cost in tilapia diets if an appropriate level is identified [31]. Due to the small
US
number of studies on the inclusion of soybean oil in the finisher diets of tilapia, this study aimed to evaluate varying levels of soybean oil in the diet of Nile tilapia and to evaluate its effects on
AN
performance parameters, hematological parameters, fillet composition and fatty acids
M
incorporation of the Nile tilapia during the finisher phase.
Materials and methods
2.1.
Experimental design and diets
ED
2.
PT
This study was conducted at the Instituto de Pesquisa em Piscicultura Ambiental (InPAA), at Universidade Estadual do Oeste do Paraná campus Toledo-PR, Brazil, and it was approved by the
CE
ethics committee for animals use in CEUA at the same institution under protocol No. 36/14. The experiment was conducted for 50 days, in which 270 Nile tilapia (O. niloticus) with an initial average weight of 425.33 ± 32.37 g and 25.53 ± 2.00 cm of length were randomly distributed in 15
AC
tanks with dimensions 2.0x6.0x0.6 m, with a capacity of 7.2 m3. The fish were fed until apparent satiation, twice a day during the finishing phase. A completely randomized design was used with five treatments and three replications. The treatments consisted of soybean oil inclusion in the diet a (0.0; 15.0; 30.0; 45.0 and 60.0 g kg−1). The diets (Table 1 and Table 2) were formulated to contain around 303.80 g kg −1 of crude protein and from 12.92 to 14.34 MJ kg−1 of digestible energy, with estimated values based on the suggested values for Nile tilapia (O. niloticus) by the [48]. The increase in soybean oil content was made at the expense of cornmeal in the diets. The addition of vitamins and minerals occurred
ACCEPTED MANUSCRIPT through a commercial premix, which meets the requirements of the species.
Table 1: Composition and nutritional diets with different soybean oil inclusions Oreochromis niloticus. Soybean oil inclusion g kg−1 15.0
30.0
45.0
60.0
Corn
254.1
236.7
219.4
202.0
184.6
Soybean meal 46%
120.9
123.3
125.8
130.6
Wheat middling
150.0
150.0
IP
128.2
150.0
150.0
150.0
Wheat bran
169.5
169.0
168.6
167.6
Bone and meat meal 42%
24.0
24.9
25.3
25.8
26.30
Poultry by-product meal 58%
200.0
200.0
CR
168.1
200.0
200.0
200.0
Feather meal 84%
60.0
60.0
60.0
60.0
60.0
Soybean oil
0.00
15.0
30.0
45.0
60.0
Choline chloride 75%
1.50
1.50
1.50
1.50
1.50
Sodium chloride
3.00
3.00
3.00
3.00
3.00
DL-Methionine
2.70
2.70
2.70
2.80
2.80
L-lysine 50.7%
6.30
6.20
6.10
6.00
6.00
2.20
2.20
2.20
2.20
2.20
0.40
0.40
0.40
0.40
0.40
0.10
0.10
0.10
0.10
0.10
5.0
5.0
5.0
5.0
5.0
0.34
0.35
0.36
0.36
0.37
PT
Vitamin C Antioxidant 1
CE
Vitamin and mineral supplement for fish 2
AC
Cost of diets (US$ kg−1)
Moisture
US AN
ED
L-Threonine
T
0.00
M
Ingredients g kg−1
−1
Analysed values (g kg ) 76.53
76.49
77.03
76.87
76.32
Crude protein
302.49
303.89
303.18
302.98
303.67
Crude fiber
40.16
41.75
40.92
40.15
41.07
Crude fat
49.31
66.22
79.82
93.91
111.22
Ash
75.97
77.34
77.97
75.34
77.35
Calcium
12.37
13.88
14.03
12.96
13.98
Phosphorus
11.53
11.77
12.67
12.12
12.34
ACCEPTED MANUSCRIPT Calculated values (g kg−1) 18.00
18.00
18.00
18.00
18.00
DL-Methionine
7.50
7.50
7.50
7.50
7.50
Methionine + cystine
13.10
13.10
13.10
13.10
13.10
Arginine
20.30
20.30
20.30
20.30
20.30
Threonine
13.50
13.50
13.50
13.50
13.50
Tryptophan
3.10
3.10
3.10
3.10
3.10
Choline g kg−1
3.00
3.00
3.00
3.00
3.00
Fish digestible energy MJ kg−1
12.92
13.25
13.96
14.34
Fish digestible protein
262.00
262.00
262.00
262.00
262.00
Starch
296.20
284.00
271.80
259.60
247.50
IP
CR
1
13.63
BHT (butylated hydroxytoluene).
2
US
*All ingredients were bought at local market.
T
Lysine
Supplementation levels per kg of feed: Vit. A 12,000 IU; Vit. D3 3000 IU; Vit. K3 15 mg kg −1; Vit.
AN
B1 20 mg kgVit. B2 20 mg kg−1; Vit. B6 18 mg kg−1; Vit. B12 0.04 mg kg−1 ; Vit C 300 mg kg−1; Niacin 100 mg kg−1; Calcium Pantothenate 50 mg kg−1; Biotin 1 mg kg−1; Folic Acid 6 mg kg−1;
M
Inositol 150 mg kg−1; Choline 500 mg kg−1; Sulf. Copper 18 mg kg−1; Sulf. Iron 80 mg kg−1; Sulf. Manganese 50 mg kg−1; Sulf. Zinc 120 mg kg−1; Calcium iodate 0.8 mg kg−1; Sulf. Cobalt 0.6 mg
ED
kg−1; selenium 4 mg kg−1, Vit. E 200 mg kg−1. 3 Based on available nutrients (NRC, 2011). and
PT
calculated by Super Crac Software (TD Software, Viçosa, MG, Brazil.
Fatty acid (mg g−1)
0.00
Soybean oil inclusion g kg−1 15.0
30.0
60.0
2.61 ±0.08
205.51 ±4.41
115.96 ±2.41
64.88 ±1.41
61.16 ±1.35
61.44 ±1.36
63.33 ±1.41
62.88 ±1.39
16:1n-9
0.69 ±0.03
0.30 ±0.02
0.20 ±0.01
0.13 ±0.01
0.13 ±0.01
16:1n-7
1.75 ±0.09
1.41 ±0.07
1.49 ±0.08
1.49 ±0.08
1.46 ±0.08
18:1n-9
171.52 ±4.32
207.00 ±5.22
203.26 ±5.21 204.65 ±5.16
207.99 ±5.24
18:1n-7
33.51 ±1.18
23.54 ±0.82
18:2n-6
295.3 ±6.94
361.47 ±8.51
16:0 18:0
2.48 ±0.08
45.0
5.48 ±0.18
AC
14:0
CE
Table 2: Fatty acid composition (mg g−1) of total lipids of experimental diets.
2.45 ±0.08
2.34 ±0.08
116.35 ±2.44 116.37 ±2.44
116.45 ±2.44
23.82 ±0.84
11.27 ±0.40
11.36 ±0.41
362.99 ±8.54 368.48 ±8.67
378.71 ±8.91
ACCEPTED MANUSCRIPT 18:3n-3
27.58 ±0.34
39.20 ±0.48
41.56 ±0.51
43.45 ±0.53
44.95 ±0.55
n 6 / n3
10.71 ±0.14
9.22 ±0.11
8.73 ±0.11
8.48 ±0.10
8.43 ±0.10
SFA
275.87 ±5.81
179.73 ±3.81
180.27 ±3.83 182.15 ±3.87
181.67 ±3.86
MUFA
207.47 ±4.59
232.24 ±5.14
228.77 ±5.06 217.53 ±4.81
220.95 ±4.88
PUFA
322.88 ±7.50
400.67 ±9.30
404.55 ±9.39 411.93 ±9.57
423.66 ±9.84
806.22 ±12.28
812.63 ±12.37
813.58 ±12.39 811.6 ±12.36 826.27±12.59
T
IP
*Results expressed as mean ± standard deviation of three replicates; values with different letters on the same line are significantly different (Tukey) P < 0.05 ; LA: linoleic acid; AA: arachidonic
CR
acid; LNA: -linolenic acid; EPA: eicosapentaenoic acid; DHA: docosahexaenoic acid; SFA: total saturated fatty acids; MUFA: total of monounsaturated fatty acids; PUFA: total
US
polyunsaturated fatty acids; n-6: total of omega 6 fatty acids; n-3: total omega-3 fatty acids; n-6/n-3: rate of omega 6/omega 3; PUFA/SFA: total polyunsaturated fatty acids/total saturated
AN
fatty acids.
M
For the development of diets, the feeds were individually milled in a hammer mill (Vieira, MCS 280, Tatuí-São Paulo, Brazil) with a 0.5 mm sieve. After milling, the ingredients were
ED
weighed and mixed manually. For the inclusion of the different levels of soybean oil in the diet, a premix of the oil was made with a sample of the other ingredients and then mixed with the rest of
PT
the diet. Afterwards, the whole mixture was moistened with 22% of water and processed through the extrusion process with a 3.0 mm matrix through an extruder (Ex-Micro® extruder, ExTeec
CE
Company, Ribeirão Preto, Brazil), keeping the temperature at 80°C, o ensure the diets were not overheated. The diets were dried out in a forced air oven (ET-394/3 Tecnal Piracicaba, Brazil) for
2.2.
AC
12 hours at 55°C.
Water parameters
The physical and chemical water parameters, such as dissolved oxygen (mg L−1), pH and electrical conductivity (µS cm−1) were measured weekly, while the temperature (°C) was measured twice a day with the assistance of the YSI Professional Plus Multiparameter Water Quality Meter device (YSI Pro Plus, Yellow Springs, Ohio, USA). The physical and chemical parameters of the water were on average 7.5 ± 0.83 mg O2 L−1; 6.98 ± 0.86 and 33.42 ± 14.64 µS cm−1 for dissolved oxygen, pH and electrical conductivity, respectively.
ACCEPTED MANUSCRIPT
2.3.
Zootechnical parameters
At the end of the trial, the fish were fasted for 24 hours, to ensure gastrointestinal tract was empty, and then were euthanized with a water benzocaine solution at 250 mg L−1 [39]. The fish were weighed, measured and counted to determine the performance parameters. The weight gain (initial
T
body weight - final body weight), daily weight gain (weight gain/day), survival [(final number of
IP
fish / initial number of fish) x100] and feed conversion (dry diet feed (g) / wet weight gained (g)) were evaluated. The Diet cost rate was calculated, following DCR = diet cost rate (per kg) / weight
CR
gained. Nine fish in each experimental unit were used for the carcass yield assessments ([(body weight (g) - visceral fat weight (g))/(body weight (g))x100]), dressed out yield ([(body weight (g)
US
- (visceral weight (g) + fish head(g))) / (body weight (g)) x100]) and fillet yield ((fillet weigh (g)/body weight (g))x100), hepatosomatic index ([liver weight (g)x100 / final body weight (g)])
Biochemical parameters of blood
M
2.4.
AN
and visceral fat ([visceral fat weight (g) x100 / final body weight (g)]).
ED
Three fish were selected from each experimental unit (nine fish per treatment), for the analysis of hematological and biochemical blood parameters. The fish were anesthetized with benzocaine (100 mg L−1) following the methods described by [50]. An erythrocyte count was performed in a
PT
Neubauer chamber using the microhematocrit method according to [27], and hemoglobin by the cyanmethaemoglobin method [14]. For biochemical blood parameters, plasma was analyzed for
CE
the following parameters: plasma glucose, total serum proteins, total cholesterol, and triglycerides were evaluated using enzymatic-colorimetric methods with the aid of specific kits for each analyte
2.5.
AC
(Gold analyzes Diagnostica, Belo Horizonte - Minas Gerais, Brazil).
Chemical analysis
For the chemical composition analysis of the diets and fillets, the methodology proposed by the [1] was used in which the moisture content was determined by drying the sample in an air circulation oven forced to 105°C until constant weight was reached (Method number 950.46), The ashes were determined by calcination of samples in a muffle furnace (Model 2000B, Belo Horizonte - Minas Gerais, Brazil) at 600°C method number 920.153, calcium and phosphorus were determined by
ACCEPTED MANUSCRIPT method number 968.08D, crude fiber method number 962.09 and protein by the Kjeldahl method (Model MA-036, Piracicaba - São Paulo, Brazil) method number 981.10. The total lipids were determined following [7] methodology. All analyses were performed in triplicate. Fatty acids derivatization was performed following the methodology described by [21]. Fatty acid methyl esters (FAME) were separated by gas chromatography (TRACE Ultra Thermo Scientific (Thermo Scientific, USA) equipped with flame ionization detector (FID) and a fused
T
silica capillary column (100 m x 0.25 mm i.d., 0.25 µm cyanopropyl, CP-7420 select FAME).
IP
Retention times and peak areas were determined using Chrom-Quest software (Thermo Scientific,
Statistical analysis
US
2.6.
CR
USA), following equipment cookbook and [59].
A one-way analysis of variance (ANOVA) was carried out with each diet as treatments and each
AN
tank as experimental units, the data were submitted to the Shapiro-Wilk normality test, at 5% probability. When significant differences were observed, the means were compared using Tukey’s
M
test at a 5% significance level. Regression analyses were performed to estimate the relationship between the treatments and responses. When more than one of the models showed significant
ED
coefficients, the regressions with the best biological explanation were chosen. All data were
Results
CE
3.
PT
analyzed using the software package R [53].
The results for growth performance at the end of the experiment are summarized in Table 3. There
AC
was a quadratic effect ( P < 0.05 ) based on increasing soybean oil inclusion on the variables daily weight gain, weight gain, final weight, carcass yield, visceral fat index, and hepatosomatic index. There was no effect ( P > 0.05 ) of soybean oil inclusion on feed conversion, fillet yield, dressed out yield and survival. Figure 1: Zootechnic parameters of Soybean oil inclusion in Tilapia’s nutrition
The optimal levels of inclusion of soybean oil in diets for Nile tilapia were determined by the adjustments of the quadratic regression models applied to the performance variables. The
ACCEPTED MANUSCRIPT inclusion of 45.29 g kg−1 (Figure 1a) and 54.36 g kg−1 of soybean oil provided better daily weight gain and higher final tilapia weight as shown in Figures S1a and S1b. The best results for carcass yield (Figure 1b), visceral fat and hepatosomatic index were observed with the inclusion of 37.60 kg−1, 55.17 kg−1 and 30.41 kg−1 of soybean oil respectively, as we can see in Figure S3a. The cost of the diet was lowest with the inclusion of 36.90 kg −1 of soybean oil (Figure 1c).
T
Table 3: Performance of fish fed with diets with increasing levels of soybean oil. Soybean oil inclusion g kg-1
SEM
p-value
Effect
407.33
409.00
0.64
>0.05
-
± 8.07
± 8.11
± 8.14
3.99 ±
4.11 ±
4.36 ±
4.17 ±
0.06
< 1.00
Quadratic
0.06 d
0.08 c
0.08 b
0.08 a
0.08 b
1.74 ±
1.76 ±
1.76 ±
1.77 ±
1.76 ±
0.03
0.03
0.03
0.03
0.03
Weight gain
185.66
199.66
205.56
218.00
208.33
(g) 3
± 2.99 d
± 3.198 ± 3.29 b
± 3.49
± 3.33 b
15.0
30.0
45.0
404.66
404.00
405.66
± 8.05
± 8.04
Daily weight
3.71 ±
gain (g) 1 Feed
590.33
(g)
± 9.45 d
611.33
625.33
617.33
± 9.65
± 9.78
± 10.00
± 9.88
cd
bc
b
ab
1.84 ±
1.76 ±
1.74 ±
1.67 ±
1.78 ±
(103) (US$/g)
0.03 a
0.02 b
0.02 bc
0.02 c
0.03 ab
AC
Diet cost rate
Carcass yield (%) 5 Dressed out yield (%) 6 Fillet yield
0.01
>0.05
-
2.87
< 1.00
Quadratic
88.27 ± 89.55 ± 89.93 ± 90.09 ± 89.44 ± 0.19 c
0.20 b
0.20 ab
0.20a
0.17
0.17
0.16
3.15
< 1.00
Quadratic
x106
3,12
< 1.00
Quadratic
x106
0.18
< 1.00
Quadratic
x106
0.20 b
51.18 ± 52.51 ± 52.10 ± 49.46 ± 50.17 ± 0.17
x106
a
603.66
CE
Final weight
PT
c
ED
rate 2
4
x106
M
conversion
AN
Initial weight
US
0.00
CR
60.0
IP
Parameters
0.48
>0.05
-
0.20
>0.05
-
0.16
33.72 ± 34.42 ± 34.00 ± 33.66 ± 33.80 ±
ACCEPTED MANUSCRIPT 0.09
0.08
0.08
0.08
Visceral fat
4.82 ±
4.19 ±
4.11 ±
4.01 ±
3.84 ±
index (%) 8
0.19 a
0.17 b
0.17 b
0.16 b
0.15 b
Hepatosomatic
3.66 ±
2.86 ±
3.05 ±
3.03 ±
3.49 ±
index (%) 9
0.24 a
0.198 c
0.20 bc
0.20bc
0.23 ab
100
100
100
100
100
Survival (%) 10
[*] 1 Daily weight gain [weight gain/day];
0.10
0.0003
Quadratic
0.10
0.0042
Quadratic
-
-
-
T
0.08
2
feed conversion [dry diet feed (g)/wet weight gained 4
Diet cost rate [cost of feed (per
CR
(g)]; 3 weight gain [final body weight initial body weight];
IP
(%) 7
kg)/weight gained x 1000]; 5 carcass yield assessments [(body weight (g) visceral fat weight
7
head(g)))/(body weight (g))x100];
US
(g))/(body weight (g))x100]; 6 dressed out yield [(body weight (g) (visceral weight (g) fish fillet yield [(fillet weigh (g)/body weight (g)) x100];
weight (g) x 100/final body weight (g)];
10
9
hepatosomatic index [liver
AN
visceral fat [visceral fat weight (g) x 100/final body weight (g)];
8
survival [(final number of fish/initial number of fish) x
M
100].
ED
Equation
gain
+ 2.50
kg-1)
R2
0
1
1
0.8929
0.0114
0.0009
< 1.00
PT
Y = 3.69
Adjusted
46.30
SEE
RPE
p-value
0.0010
< 1.00
x106
CE
Daily weight
SO (g
x106
x102 0SO
AC
3.00
x104 SO
2
Weight gain
Y = 184.4 + 1.25SO
1.39 x102 SO 2
45.29
0.8812
0.5699
0.0450
0.0007
0.0516
< 1.00
x106
ACCEPTED MANUSCRIPT Final weight
Y = 590.5
54.36
0.8264
0.8570
0.0677
0.0011
0.0385
< 1.00
x106
+ 1.11SO
1.02 x102 SO 2
Diet cost rate Y = 18.5
36.90
0.8377
0.0137
37.60
0.9183
0.1060
0.01081
0.00173
x102 SO
+ 9.70
1.30
3.20 x102 SO + 3.00
2
55.17
CE
x104 SO
Y = 3.58
ic index
4.50
AC
Hepatosomat
x106
x106
0.7518
0.0916
0.0072
< 1.00
0.0073
0.0003
0.0159
0.00416
x106
PT
index
ED
2
Y = 4.75
< 1.00
M
x103 SO
Visceral fat
0.0084
AN
x102 SO
US
2
yield
< 1.00
CR
+ 1.13
Y = 88.3
0.0004
IP
x101 SO
Carcass
0.00163
T
8.34
(10 3 )
0.0003
30.41
0.6860
0.1151
0.0091
0.0002
x102 SO
+ 7.00
x104 SO 2
* SO = Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE = Relative prediction error
ACCEPTED MANUSCRIPT
Results of hematological and biochemical are summarized in Table 4. For hematological and biochemical blood variables there was an effect ( P < 0.05 ) of increasing soybean oil levels on plasmatic cholesterol and triglycerides. The diet inclusion of soybean oil had no effect ( P > 0.05 ) on glucose, plasma proteins, hemoglobin, and total red cell hematocrit percentage for the blood
IP
T
parameters.
soybean oil. Parameters
CR
Table 4: Haematology and biochemical parameters of fish fed with diets with increasing levels of
Soybean oil inclusion g kg-1 15.0
30.0
45.0
56.04 ±
58.85 ±
58.07 ±
54.65 ±
55.69 ±
8.43
8.85
8.73
8.21
8.37
4.10 ±
4.31 ±
4.22 ±
0.17
0.17
0.17
9.18 ±
8.26 ±
0.84
Erythrocyte
3.97 ±
0.16
0.16
7.56 ±
8.68 ±
9.45 ±
0.75
0.69
0.80
0.88
2.20 ±
2.19 ±
2.17 ±
2.15 ±
2.33 ±
(106 mL-1)
0.10
0.10
0.10
0.10
0.10
Haematocrit
39.22 ±
39.11 ±
39.00 ±
39.11 ±
40.44 ±
0.75
0.74
0.74
0.74
0.77
Total
114.56
122.15
152.92
135.37
222.61
cholesterol
± 5.49 d
± 5.87
± 7.37 b
± 6.49 c
± 10.69
dL-1) Haemoglobin (g dL-1)
AC
(mg dL-1)
CE
(%)
ED
Protein (g
M
4.08 ±
PT
dL-1)
AN
Glucose (mg
60.0
US
0.00
cd
SEM
p-value
Effect
2.84
>0.05
-
0.05
>0.05
-
0.25
>0.05
-
0.03
>0.05
-
0.22
>0.05
-
2.38
< 1.00
Quadratic
x106
a
Triglycerides
184.04
205.94
220.37
268.33
269.35
(mg dL-1)
± 33.84
± 37.06
± 39.67
± 2.30
± 48.48
b
ab
ab
a
a
11.02
0.0143
-
Equation SO (mg−1
Adjusted
SEE
RPE
p-value
ACCEPTED MANUSCRIPT
Total
Y=
g)
R2
0
1
2
8.30
0.7533
3.3396
0.2637
0.0042
0.012303
0.000367
cholesterol 119.5002
0.5844SO + 0.03522SO2
T
* Means with different letter in the same raw are significantly different (Tukey) P < 0.05 ; SO =
IP
Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE =
CR
Relative prediction error
US
According to the quadratic regression analysis (Table4 and Figure S2b), the lowest plasma cholesterol level was observed with the addition of 8.30 g kg−1 of soybean oil in the diet. For
AN
triglycerides, statistical differences are shown ( P < 0.05 ) between the diets with the lowest level being observed in the lowest inclusion level and the highest level of triglycerides being seen with the inclusion of 60 g kg−1 of soybean oil.
M
For the chemical composition of the tilapia fillets (Table 5), a difference ( P < 0.05 ) was
ED
observed between treatments for crude protein and crude fat. There was no effect ( P > 0.05) on
PT
moisture and ash.
Table 5: Composition of tilapia fillets submitted to diets with increasing levels of soybean oil.
CE
Parameters (g kg-1)
0.00
15.0
30.0
45.0
60.0
SEM
p-value
766.0 ±
766.2 ±
766.1 ±
760.7 ±
765.3 ±
0.14
> 0.05
6.05
6.05
6.05
6.07
6.05
180.5 ±
173.9 ±
171.6 ±
171.5 ±
172.3 ±
0.11
0.0085
3.30 a
3.18 ab
3.14 b
3.14 b
3.15 b
38.3 ±
40.6 ±
43.0 ±
45.0 ±
43.3 ±
0.12
< 1.00
3.85 b
4.08 ab
4.33 a
4.53 a
4.35 a
10.9 ±
10.7 ±
11.2 ±
10.7 ±
11.1 ±
0.42
0.42
0.43
0.41
0.43
AC
Moisture
Crude protein
Crude fat
Ash
Soybean oil inclusion g kg-1
x106 0.01
> 0.05
*Means with different letter in the same raw are significantly different (Tukey) P < 0.05 ; SEM =
ACCEPTED MANUSCRIPT standard error of the mean;
The highest crude protein content in fillets was observed in the treatment without oil inclusion. For crude fat, there was a lower value for the control treatment and the 15.0 kg −1 of soybean oil inclusion. However, the other treatments had higher crude fat values. Fatty acid compositions of the tilapia fillets are summarized in Table 6. A total of 22 fatty
T
acids were determined in the tilapia fillets. Among saturated fatty acids (SFA), the highest
IP
concentration was observed for 16:0 (238.40 to 262.19 mg g −1); monounsaturated fatty acids
CR
(MUFA) was 18:1n-9 (215.36 to 277.61) and polyunsaturated fatty acids (PUFA), was 18:2n-6 (LA) (157.84 to 224.49).
US
There was a quadratic effect ( P < 0.05) , (Table6 and Figures 2a and 2b) when dietary soybean oil level increased for 16:1n-7; 18:3n-6, 18:3n-3 (LNA); 20:2n-6 and the total of omega 3
AN
( n-3) fatty acids.
M
Table 6: Fatty acid composition (mg g−1) of total lipids of fillet from Oreochromis niloticus after being fed diets with different soybean oil inclusions. Soybean oil inclusion g kg 1
14:0
0.00
15.0
30.0
45.0
60.0
SEM
p-value
Effect
29.57 ±
31.17 ±
30.34 ±
29.64 ±
35.01±
0.085
>0.05
-
3.00
2.92
2.85
3.37
3.73 ±
3.78 ±
4.07 ±
3.85±
0.054
2.56
-
0.06 c
0.06 b
0.07 c
2.85 4.25 ±
CE
15:0
0.06 a
17:0
18:0
16:1n-7
262.19 ±
AC
16:0
PT
(mg g−1)
ED
Fatty acid
0.05 c
238.40 ± 241.70 ± 260.19 ± 246.30 ±
38.80 a
35.28 b
35.77 b
38.51 a
36.45 b
2.39 ±
3.12 ±
6.68 ±
5.85 ±
3.82 ±
0.15 d
0.19 c
0.41 a
0.36 b
0.24 c
66.30 ±
65.99 ±
54.76 ±
47.14 ±
66.60 ±
6.62 a
4.37 a
2.39 b
1.13 c
9.99 a
68.21±
68.71 ±
62.26 ±
33.67 ±
22.25 ±
9.55 a
9.62 a
8.72 a
4.71 b
3.11 b
x1006 0.271
2.93
-
x1005 0.045
< 1.00
-
x106 0.212
< 1.00
-
x106 0.542
4.14
x105
Quadratic
ACCEPTED MANUSCRIPT 25.03 ±
1.17 a
0.83 bc
0.81 c
0.80 c
0.91 b
20:3n-6
20:4n-6 (AA)
2.46 c
2.35 d
6.88 ±
5.90 ±
5.61 ±
5.60 ±
5.33 ±
0.07 a
0.06 b
0.06 c
0.06 c
0.06 d
20.31 ±
17.45 ±
16.56 ±
16.53 ±
15.76 ±
0.22 a
0.19 b
0.18 c
0.18 c
0.17 d
< 1.00
0.014
< 1.00
0.042
2.36 a
4.61 ±
5.38 ±
6.49 ±
7.28 ±
5.61 ±
0.29 c
0.33 c
0.59 a
0.45 b
0.35 c
x106
10.24 ±
10.83 ±
12.05 ±
12.14 ±
12.08 ±
0.19 c
0.19 b
0.22 a
0.22 a
0.22 a
3.55 ±
3.92 ±
3.72 ±
3.71 ±
3.54 ±
0.09 c
0.04 a
0.04 b
0.04 b
0.04 c
6.57 ±
6.37 ±
8.08 ±
12.28 ±
16.13 ±
0.40 cd
0.39 d
0.49 c
0.75 b
0.99 a
38.06 ±
40.83 ±
43.24 ±
48.85 ±
49.28 ±
0.43 c
0.45 b
0.51 a
0.52 a
CE
27.83 ±
28.23 ±
27.73 ±
(LNA)
0.62 c
0.91 b
1.07 a
1.09 a
1.07 a
20:4n-3
2.18 ±
2.41 ±
2.32 ±
2.34 ±
2.21 ±
0.05 bc
0.06 a
0.06 abc
0.06 ab
0.06 bc
1.47 ±
1.34 ±
1.62 ±
2.37 ±
2.61 ±
(EPA)
0.11 b
0.10 b
0.13 b
0.18 a
0.20 a
22:6n-3
4.92 ±
5.41 ±
5.48 ±
5.48 ±
5.47 ±
(DHA)
0.08 b
0.09 a
0.09 a
0.09 a
0.09 a
n-6
200.87 ±
AC
23.70 ±
1.89 d
253.33 ± 273.56 ± 306.78 ± 311.12 ± 2.38 c
2.57 b
2.88 a
2.92 a
0.047
0.021
-
x106
2.34 a
16.19 ±
0.844
< 1.00
2.06 b
18:3n-3
-
x106
1.95 c
186.00 ± 196.99 ± 222.52 ± 224.49 ±
-
x106
1.66 d
0.40 d
20:5n-3
0.583
< 1.00
157.84 ±
-
x106
CR
20:2n-6
2.47 c
US
18:3n-6
2.60 b
< 1.00
AN
18:2n-6 (LA)
3.03 a
M
20:1n-9
238.46 ± 226.38 ± 225.94 ± 215.36 ±
0.103
T
21.89 ±
IP
22.23 ±
277.61 ±
18:1n-11
22:5n-6
22.62 ±
ED
18:1n-9
32.04 ±
PT
18:1n-7
-
x106 < 1.00
< 1.00
Quadratic
-
x106 0.038
2.92
-
x1006 0.102
< 1.00
-
x106 0.118
< 1.00
-
x106 0.124
< 1.00
Quadratic
x106 0.026
3.13
Quadratic
x103 0.014
2.15
Quadratic
x103 0.006
4.60
-
x1005 1.078
< 1.00
x106
-
ACCEPTED MANUSCRIPT
SFA
32.85 ±
37.26 ±
38.43 ±
38.02 ±
0.72 c
0.95 b
1.08 a
1.11 a
1.10 a
8.11 ±
7.72 ±
7.34 ±
7.98 ±
8.19 ±
0.21 a
0.20 ab
0.19 b
0.21 a
0.21 a
364.70 ±
342.41 ± 337.25 ± 346.88 ± 355.59 ±
3.90 a
MUFA
3.66 c
405.04 ±
3.71 bc
7.89 b
0.094
7.20 c
6.72 c
2.97
0.273
3.123e
1.138
-
3.26 b
3.62 a
3.66 a
PUFA/
0.62 ±
0.84 ±
0.92 ±
0.99 ±
0.98 ±
SFA
0.01 d
0.01 c
0.01 b
0.01 a
0.01 a
x106
0.037
-
x106
3.00 c
US
1.212
< 1.00
2.37 d
286.18 ± 310.82 ± 345.21 ± 349.14 ±
-
x1005
< 1.00
225.63 ±
-
x103
CR
8.37 b
< 1.00
x106
3.80 ab
353.15 ± 333.04 ± 303.63 ± 283.73 ±
9.60 a
PUFA
3.61 c
0.139
T
n-6/ n-3
24.76 ±
IP
n-3
-
x106 < 1.00
-
AN
*Results expressed as mean standard deviation of three replicates; values with different letters on the same line are significantly different (Tukey) P < 0.05 ; LA: linoleic acid; AA: arachidonic
M
acid; LNA: -linolenic acid; EPA: eicosapentaenoic acid; DHA: docosahexaenoic acid; SFA: total saturated fatty acids; MUFA: total of monounsaturated fatty acids; PUFA: total
ED
polyunsaturated fatty acids; n-6: total of omega 6 fatty acids; n-3: total omega-3 fatty acids;
saturated fatty acids. Equation
Adjusted
g−1)
R2
0
1
2
1.02
0.946
4.15
3.28x101
33.22
0.960
2.18x101 1.72 x102
CE
Y = 69.8
SO (g
AC
16:1n-7
PT
n-6/ n-3: rate of omega 6/omega 3; PUFA/SFA: total polyunsaturated fatty acids/total
SEE
RPE
p-value
5.20 x103
0.173
0.019
3.00 x104
0.024
< 1.00
2.97 x102 SO 1.46 x102 SO 2 18:3n-6
Y = 4.09
2.39 x101
x106
ACCEPTED MANUSCRIPT SO
3.60 x103 SO 2 18:3n-3
Y = 16.4
(LNA)
44.50
0.965
5.19 x101 4.10 x102
6.00 x104
0.016
< 1.00
x106
T
5.61x101 SO SO 2 Y = 2.21
29.25
0.780
3.12 x102 2.50 x103
1.13
0.923
7.95x102 6.30 x103
2.00 x104
5.30 x104
1.00 x104
0.018
3.35x103
M
(EPA)
AN
SO 2 Y = 1.39
0.043
US
1.17 x104 SO
20:5n-3
4.00 x105
CR
20:4n-3
IP
6.30 x103
ED
9.00 x104 SO
4.00 x104
PT
SO 2
*.SO = Soybean oil; SEM = standard error of the mean; SEE = Standard error of the estimate, RPE
CE
= Relative prediction error
4.
AC
Figure 2: Fatty acid parameters of fillets of Tilapia fed with Soybean oil inclusion
Discussion
For fish, as well as tilapia, lipids perform essential functions in growth, feed efficiency, healthiness, kidney and gill function, neural and visual development, reproduction, and fillet quality [38]. Vegetable oils are easily found on the market and are important sources of lipids and energy for tropical fish and can be used depending on cost and availability [58]. No fish mortality was observed during the experimental period. [8] conducted a study evaluating different levels of energy and soybean oil inclusion in diets for tilapia also found no
ACCEPTED MANUSCRIPT effect of diets on the animal’s survival. The animals in the current study showed no mortality because they were well nourished and were raised in an adequate environment for their normal development. The quadratic effect observed for daily weight gain demonstrated an optimal level of soybean oil inclusion in the diet at 46.30 g kg−1, corresponding to a weight gain of 4.20 g day−1 and a total weight gain of 210 g. The lowest diet cost rate was observed to be at the inclusion of 36.90
T
g kg−1. Weight gain showed a quadratic effect with optimal growth occurring with the addition of
IP
54.36 kg−1 g of soybean oil in the diet.
CR
[55] found that the inclusion of oil above the diet needs of the animals caused a reduction in the rate of enzymatic reactions and specific activity of the catalytic efficiency of glucose
US
6-phosphate dehydrogenase in the liver of trout. This may explain the decline in growth with the excessive oil level in the diet observed in the current paper.
AN
The feed conversion rate of diets with the high content of soybean oil was similar between treatments, showing that the energy level of the different diet did not influence the feed utilization
M
of the animals. [42] observed feed conversion values of 1.69 for fingerlings Nile tilapia in diets supplemented with 3 and 4.8% of fat. In the present study, the feed conversion rate was superior
ED
due to the animal breeding phase being different. It is known that feed conversion increases with the increasing weight of the animal, namely in the early stages, so it is natural that the feed
PT
conversion is higher in the final stages of farming. The diets evaluated in the current study were in a relationship between digestible protein
CE
and digestible energy between 18.27 and 20.28 mg DP kJ−1, and the best ratio of digestible protein to digestible energy was observed to be 18.77 mg DP kJ−1 with the inclusion of around 45.0 g kg 1
AC
of soybean oil addition. While [10] found an ideal ratio of digestible protein and digestible energy to be 20.45 mg DP kJ−1 for tilapia larvae in the sexual reversion phase. It is essential to consider the protein to energy ratio for a good development of fish, taking into account the impacts of the management and environment [9]. The utilization of an adequate level of oil in feeds for fish brings an opportunity for lower production cost as shown in the diet cost rate in the present study. The highest carcass yield was observed for the inclusion of 37.60 g kg −1 of soybean oil in the diet when using quadratic regression analysis (Table 3). Even though tilapia is a species that is good at utilizing carbohydrates, apparently there was an excess of starch, causing an increase in fat in the abdominal cavity. The diet without soybean oil inclusion provided an increase in the visceral
ACCEPTED MANUSCRIPT fat deposition; this diet contained the highest starch content. Starch may have influenced visceral fat, as increased oil content and decreased starch in the diet resulted in lower fish visceral fat. The higher proportion of carbohydrates may explain increased visceral fat deposition. According to the [48], the body fat deposition content in fish can increase when the diet is formulated with starch above the capacity of utilization as an energy source. However, for this phase of development, with a fully functional digestive tract, there is insufficient information
T
about starch inclusion limit in diets for tilapia.
IP
When it comes to other species, such as mammals, the mechanisms that are potentially
CR
responsible for the lower body fat deposition are the reduction of energy consumption by suppressing appetite and increasing energy expenditure in muscles by lipolysis [36]. Although the
US
fish mechanisms are not clearly understood, they are probably related to the inhibition of lipids absorption and even lipid oxidation [40].
AN
According to the quadratic regression equation for the hepatosomatic index, the lowest value was estimated to be at 30.41 g kg−1 of soybean oil inclusion. In a study by [37], evaluating
M
the interaction between crude fiber and soybean oil in diets for tilapia, the authors observed an increase in the hepatosomatic index, which was also observed by [3]. The hepatosomatic index is
ED
directly linked to glycogen accumulation in the liver and consequently causes an increase in liver weights [16, 45]. Therefore, in the present paper, the diet with the higher content of starch
PT
demonstrated a higher hepatosomatic index, results similar observed by [6] in their study with hybrid catfish, using similar content of starch in its diets. This fact, is most likely related to
CE
increased starch metabolism, through glycogenesis catalyzed by glycogen synthase [48] and glycogen deposition in the liver [28, 6, 64].
AC
The inclusion of soybean oil in the diet had no effect ( P > 0.05 ) on the glucose, total serum proteins, hemoglobin, erythrocyte, and hematocrit of the blood. Evaluating the interaction of the inclusion of 0 to 6% sunflower oil and linseed in diets for tilapia raised in reduced temperatures, [2] observed reductions in erythrocyte levels (2.02 to 1.76) in fish fed 4% of sunflower oil and 2% of flaxseed oil and 3.82 to 3.05 of total protein in fish fed 6% flaxseed oil. The authors concluded that diets with a low proportion of omega-6 and omega-3 fatty acids lead to a reduction in blood profile when the animals are submitted to a thermal challenge. According to the quadratic regression equation, the lowest plasma cholesterol level was observed with the addition of 8.30 kg−1 g of soybean oil inclusion, indicating an upward trend for
ACCEPTED MANUSCRIPT cholesterol as soybean oil inclusion is increased in the diet. This fact is possibly correlated with the characteristic of a body’s cholesterol retention and cholesterol being a precursor of bile salts necessary for the digestion of ingested lipids [46]. The total plasma cholesterol content may be regarded as indicative of the health of fish, when tilapia are kept in unhealthy environments reductions in plasma cholesterol levels have been observed [63]. [4] found a mean value of 120 mg dL−1 for plasma cholesterol, evaluating the
T
interaction of soybean oil and palm oil, as a lipid source, when feeding palm meal to tilapia. [52]
IP
observed that the use of vegetable oils in diets for Acanthopagrus schlegeli contributes to lowered
CR
plasma cholesterol. According to [62], excessive intake of feeds rich in fat content and cholesterol may increase its concentration in the blood. However, it is noteworthy that this molecule is a
US
precursor of hormones such as sterol hormones, which are responsible for the growth of fish, and vitamins needed for vital processes of reproduction and maintenance of health. Nevertheless, there
AN
is no consensus on cholesterol levels, and comparisons are often made with different diets and rearing systems, mainly when energy sources are used in the diet [47]. However, in this study, the
M
total plasma cholesterol contents are in agreement with those proposed by [32] for tilapia in intensive systems.
ED
The triglyceride levels show statistical differences ( P < 0.05 ) between the diets used. Similar results were observed by [20] as they studied the effect of lipid levels on growth and body
PT
composition of juvenile grass carp. Their work showed a linear increase in triglyceride content as the lipid content was increased in the feed. In our study, it is important to note that the increase of
CE
total cholesterol and total triglycerides according to the increased inclusion of soybean oil does not adversely influence the visceral fat deposition since there was a reduction of fat deposited in the
AC
viscera with the increasing inclusion of oil. In the analysis of the chemical composition of the tilapia fillets, the crude protein content the fillets found in this study was similar to those reported by [5], which evaluated the effect of the inclusion of palm oil sources for Oreochromis sp., obtaining results from 16.9 to 17, 9%. Similar results were also observed by [25], who evaluated the influence of inclusion of different fat sources (soybean oil, corn oil, linseed oil, fish oil and olive oil) on muscle composition and concentration of plasma lipoproteins for O. niloticus, obtaining crude protein values from 16.28 to 18.27%. The values vary according to the inclusion of oils. Animals fed with diets isonitrogenous, isocaloric, and with no changes in lipid levels
ACCEPTED MANUSCRIPT usually do not differ in the basic chemical composition [41]. However, [26] stated that the variation in chemical composition of fish might occur due to endogenous and exogenous factors such as genetics, size, sex, reproductive stage, environmental factors, temperature and season, in addition to nutrient intake. In addition, there may be a higher concentration of fat/protein in different parts of the body, in order to eliminate this factor, the fish fillets were ground and homogenized for chemical analysis in the present study.
T
Lipid nutrition is directly related to the quality of fish fillet [34], which influences the
IP
commercial value of the fish. Generally, in older fish, there is greater fat deposition compared to
CR
young fish [49], this is due to the younger fish having a higher growth rate in which dietary energy and protein is used for growth rather than storage for energy reserves [15]. Evaluating the effect of
US
soybean oil to fish oil ratio in Acanthopagrus schlegeli diets, [52] observed differences in the lipid composition of the fillets and found that soybean oil positively influences the deposition of
AN
intramuscular fat as well as decreases intraperitoneal fat deposition. Production of high-quality meat products has been a big focus lately [18]. Optimal finisher
M
diet formulations not only provide good feed efficiency and growth rate, they reduce costs, and also affect the carcass and fillet quality. The composition of the diets in this study had a direct
ED
effect (Table 6 and Figure 2a and 2b) on fillet composition and fatty acid composition. Nile tilapia metabolism requires n-6 fatty acids in their diets to maximize development. They also tend to store
PT
large amounts of these compounds in their musculature [29, 25], this propriety can be used to produce healthier products for human consumption [61].
CE
The increase of long-chain polyunsaturated essential fatty acid levels, (20:5-n3 (EPA) and (22:6-n3 (DHA)), may be attributed to the concentration -linolenic acid in, a precursor of the
AC
n-3 fatty acid series, in the diets. [12], studied the inclusion of -linolenic acid in the nutrition of Nile tilapia. They attribute the increase of DHA to the bioconversion of -linolenic acid, as a substrate, of the -6/-5 desaturases and elongase enzymes in the metabolism of Nile tilapia. Also, [44] attributed the increase of PUFAs in carcasses to the antioxidant activity of vitamin E concentrations incorporated in diets. The diets evaluated in our current work have higher levels of soybean oil that contains a great quantity of vitamin E and PUFAs [13], this can provide greater nutritional values to the fillets. All fillets in this study showed higher PFUA to SFA ratios than the recommended minimum by the Department of Health and Social Security [17].
ACCEPTED MANUSCRIPT 5.
Conclusion
It is concluded that diets with soybean oil inclusion around 45.0 g kg −1, provide better growth performance of tilapia during the finishing phase. Dietary supplementation of soybean oil was effective at improving nutrition values of the meat of tilapia as well as achieving above the recommended PUFA to SFA ratio values described by the Department of Health and Social
References
CR
6.
IP
T
Security.
References
AOAC (2005). Association of Official Analytical Chemists (AOAC). (19th ed.). Hoboken,
US
[1]
NJ, USA: John Wiley & Sons, Inc. doi:10.1002/0471740039.vec0284. Araujo, D., Pezzato, A. C., Barros, M. M., Pezzato, L. E., & Nakagome, F. K. (2011).
AN
[2]
Hematologia de tilápias-do-nilo alimentadas com dietas com óleos vegetais e estimuladas
M
pelo frio. Pesquisa Agropecuária Brasileira, 46, 294–302. doi:10.1590/s0100-204x2011000300010. Atwood, H. L., Tomasso, J. R., Webb, K., & Gatlin, D. M. (2003). Low-temperature
ED
[3]
tolerance of nile tilapia, oreochromis niloticus: effects of environmental and dietary
PT
factors. Aquaculture Research, 34, 241–251. doi:10.1046/j.1365-2109.2003.00811.x. de Azevedo, R. V., Tonini, W. C. T., & Braga, L. G. T. (2013). Óleo e torta de dend^e em
CE
[4]
rações para juvenis de tilápia-do-nilo. Pesquisa Agropecuária Brasileira, 48, 1028–1034.
[5]
AC
doi:10.1590/s0100-204x2013000800031. Bahurmiz, O. M., & Ng, W.-K. (2007). Effects of dietary palm oil source on growth, tissue fatty acid composition and nutrient digestibility of red hybrid tilapia, oreochromis sp., raised from stocking to marketable size. Aquaculture, 262, 382–392. doi:10.1016/j.aquaculture.2006.11.023. [6]
Bernardes, C.,, Navarro, R., Santos, B. G., Silva, R. F.,, & and (2016). Effects of dietary carbohydrate/lipid ratios on growth, body composition, and nutrient utilization of hybrid catfish (pseudoplatystoma reticulatum x leiarius marmoratus). Revista Colombiana de Ciencias Pecuarias, 29. URL:
ACCEPTED MANUSCRIPT https://doi.org/10.17533%2Fudea.rccp.v29n1a08. doi:10.17533/udea.rccp.v29n1a08. [7]
Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Canadian journal of biochemistry and physiology, 37, 911–917.
[8]
Boscolo, W. R., Hayashi, C., & Meurer, F. (2002). Digestibilidade aparente da energia e
[9]
IP
niloticus, l.). Revista Brasileira de Zootecnia, 31, 539–545.
T
nutrientes de alimentos convencionais e alternativos para a tilápia do nilo (oreochromis
Boscolo, W. R., Hayashi, C., & Meurer, F. (2002). Farinha de varredura de mandioca
l.). Revista Brasileira de Zootecnia, 31, 546–551.
Boscolo, W. R., Hayashi, C., Meurer, F., Feiden, A., & Wolff, L. (2004). Desempenho e
US
[10]
CR
(manihot esculenta) na alimentação de alevinos de tilápia do nilo (oreochromis niloticus
características de carcaça de tilápias do nilo (oreochromis niloticus l.) alimentadas com
AN
rações contendo diferentes níveis de gordura. Acta Scientiarum. Animal Sciences, 26, 443–447.
Boscolo, W. R., Signor, A., Feiden, A., Bombardelli, R. A., Signor, A. A., & Reidel, A.
M
[11]
(2005). Energia digestível para larvas de tilápia-do-nilo (oreochromis niloticus) na fase de
ED
reversão sexual. Revista Brasileira de Zootecnia, 34, 347 1813–1818. doi:10.1590/s1516-35982005000600003. Chen, C., Guan, W., Xie, Q., Chen, G., He, X., Zhang, H., Guo, W., Chen, F., Tan, Y., &
PT
[12]
Pan, Q. (2018). n-3 essential fatty acids in nile tilapia, oreochromis niloticus :
CE
Bioconverting LNA to DHA is relatively efficient and the LC-PUFA biosynthetic pathway is substrate limited in juvenile fish. Aquaculture, 495, 513–522.
[13]
AC
doi:10.1016/j.aquaculture.2018.06.023. Clemente, T. E., & Cahoon, E. B. (2009). Soybean oil: Genetic approaches for modification of functionality and total content. PLANT PHYSIOLOGY, 151, 1030–1040. doi:10.1104/pp.109.146282. [14]
Collier, H. B. (1944). Standardization of blood haemoglobin determinations. Canadian Medical Association Journal, 50, 550.
[15]
Contreras-Guzmán, E. S. (1994). Bioquímica de pescados e derivados. FUNESP.
[16]
Cyrino, J. E. P., Portz, L., & Martino, R. C. (2000). Retenção de proteína e energia em juvenis de. Scientia Agricola, 57, 609–616.
ACCEPTED MANUSCRIPT [17]
DHSS, U. (1994). Nutritional Aspects of Cardiovascular Disease. Technical Report Report on Health and Social Subjects London.
[18]
Dokmanovic, M., Baltic, M. Z., Duric, J., Ivanovic, J., Popovic, L., Todorovic, M., Markovic, R., & Pantic, S. (2015). Correlations among stress parameters, meat and carcass quality parameters in pigs. Asian-Australasian Journal of Animal Sciences, 28, 435–441. doi:10.5713/ajas.14.0322.
T
Drotningsvik, A., Mjøs, S. A., Pampanin, D. M., Slizyte, R., Carvajal, A., Remman, T.,
IP
[19]
Høgøy, I., & Gudbrandsen, O. A. (2016). Dietary fish protein hydrolysates containing
CR
bioactive motifs affect serum and adipose tissue fatty acid compositions, serum lipids, postprandial glucose regulation and growth in obese zucker fa/fa rats. British Journal of
[20]
US
Nutrition, 116, 1336–1345.
Du, Z.-Y., Liu, Y.-J., Tian, L.-X., Wang, J.-T., Wang, Y., & 373 Liang, G.-Y. (2005).
AN
Effect of dietary lipid level on growth, feed utilization and body composition by juvenile grass carp (ctenopharyngodon idella). Aquaculture Nutrition, 11, 139–146. Duckett, S. K., Andrae, J. G., & Owens, F. N. (2002). Effect of high-oil corn or added corn
M
[21]
oil on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in
ED
beef steers fed finishing diets. Journal of Animal Science, 80, 3353–3360. doi:10.2527/2002.80123353x. FAO (2014). The State of World Fisheries and Aquaculture 2014. Rome.
[23]
FAO (2016). The state of world fisheries and aquaculture 2016. Technical Report FAO
PT
[22]
[24]
CE
Rome. URL: http://www.fao.org/3/a-i5555e.pdf. FAO (2018). The state of world fisheries and aquaculture 2018: meeting the sustainable
[25]
AC
development goals. Rome: UN. Ferreira, M. W., de Araujo, F. G., Costa, D. V., Rosa, P. V., Figueiredo, H. C., & Murgas, L. D. (2011). Influence of dietary oil sources on muscle composition and plasma lipoprotein concentrations in nile tilapia, oreochromis niloticus. Journal of the World Aquaculture Society, 42, 24–33. [26]
Fonseca-Madrigal, J., Karalazos, V., Campbell, P., Bell, J. G., & Tocher, D. R. (2005). Inuence of dietary palm oil on growth, tissue fatty acid compositions, and fatty acid metabolism in liver and intestine in rainbow trout (oncorhynchus mykiss). Aquaculture Nutrition, 11, 241–250.
ACCEPTED MANUSCRIPT [27]
Goldenfarb, P. B., Bowyer, F. P., Hall, E., & Brosious, E. (1971). Reproducibility in the hematology laboratory: the microhematocrit determination. American Journal of Clinical Pathology, 56, 35–39.
[28]
Hemre, G.-I., Monnsen, T., & Krogdahl, A. (2002). Carbohydrates in fish nutrition: effects on growth, glucose metabolism and hepatic enzymes. Aquaculture Nutrition, 8, 175–194. URL: https://doi.org/10.1046%2Fj.1365-2095.2002.00200.x.
IP
[29]
T
doi:10.1046/j.1365-2095.2002.00200.x.
Higuchi, L. H., Feiden, A., Matsushita, M., Santarosa, M., Zanqui, A. B., Bittencourt, F., &
CR
Boscolo, W. R. (2013). Quantificação de ácidos graxos de alevinos de tilápia do nilo (oreochromis niloticus) alimentados com diferentes fontes de óleos vegetais. Semina:
[30]
US
Ciências Agrárias, (pp. 1913–1924).
Hildsorf, A. (1995). Genética e cultivo de tilápias vermelhas, uma revisão. Boletim do
[31]
AN
Instituto de Pesca, 22, 73–84.
Hooley, C. G., Barrows, F. T., Paterson, J., & Sealey, W. M. (2014). Examination of the
M
effects of dietary protein and lipid levels on growth and stress tolerance of juvenile tilapia,oreochromis niloticus. Journal of the World Aquaculture Society, 45, 115–126.
[32]
ED
doi:10.1111/jwas.12110.
Hrubec, T. C., Cardinale, J. L., & Smith, S. A. (2000). Hematology and plasma chemistry
PT
reference intervals for cultured tilapia (oreochromis hybrid). Veterinary clinical pathology, 29, 7–12.
Izquierdo, M., Obach, A., Arantzamendi, L., Montero, D., Robaina, L., & Rosenlund, G.
CE
[33]
(2003). Dietary lipid sources for seabream and seabass: growth performance, tissue
[34]
AC
composition and flesh quality. Aquaculture Nutrition, 9, 397–407. Justi, K., Hayashi, C., Visentainer, J., De Souza, N., & Matsushita, M. (2003). The influence of feed supply time on the fatty acid profile of nile tilapia (oreochromis niloticus) fed on a diet enriched with n-3 fatty acids. Food chemistry, 80, 489–493. [35]
Kaushik, S. (1990). Importance des lipides dans la alimentation des poissons. Aqua Revue, 29, 9–16.
[36]
Kennedy, A., Martinez, K., Schmidt, S., Mandrup, S., LaPoint, K., & McIntosh, M. (2010). Antiobesity mechanisms of action of conjugated linoleic acid. The Journal of nutritional biochemistry, 21, 171–179.
ACCEPTED MANUSCRIPT [37]
Lanna, E. A. T., Pezzato, L. E., Furuya, W. M., Vicentini, C. A., Cecon, P. R., & Barros, M. M. (2004). Fibra bruta e óleo em dietas práticas para alevinos de tilápia do nilo (oreochromis niloticus). Revista Brasileira de Zootecnia, (pp. 2177–2185).
[38]
Lim, C., & Webster, C. D. (2001). Nutrition and fish health. Food Products Press New York.
[39]
Luna, S., & Teixeira, M. (2007). Eutanásia: considerações éticas e indicações técnicas.
Makol, A., Torrecillas, S., Fernández-Vaquero, A., Robaina, L., Montero, D., Caballero,
IP
[40]
T
Revista CFMV, 13, 60–69.
CR
M. J., Tort, L., & Izquierdo, M. (2009). Effect of conjugated linoleic acid on dietary lipids utilization, liver morphology and selected immune parameters in sea bass juveniles
and Molecular Biology, 154, 179–187.
Menoyo, D., Lopez-Bote, C. J., Bautista, J. M., & Obach, A. (2003). Growth, digestibility
AN
[41]
US
(dicentrarchus labrax). Comparative Biochemistry and Physiology Part B: Biochemistry
and fatty acid utilization in large atlantic salmon (salmo salar) fed varying levels of n-3 and
[42]
M
saturated fatty acids. Aquaculture, 225, 295–307. Meurer, F., Hayashi, C., Boscolo, W. R., & Soares, C. M. (2002). Lipídeos na alimentação
Zootecnia, 31, 566–573.
Moreira, P. O., Rocha, J. D. M., da Silva, T. C., Bittencourt, F., Dacley Hertes, N., &
PT
[43]
ED
de alevinos revertidos de tilápia do nilo (oreochromis niloticus l.). Revista Brasileira de
Boscolo, W. R. (2015). Óleo de soja utilizado pré e pós-processamento de rações para
[44]
CE
tilápia do nilo. Bol. Inst. Pesca, São Paulo, 41, 547–555. Navarro, R. D., Navarro, F. K. S. P., Filho, O. P. R., Ferreira, W. M., Pereira, M. M., &
AC
Filho, J. T. S. (2012). Quality of polyunsaturated fatty acids in nile tilapias (oreochromis niloticus) fed with vitamin e supplementation. Food Chemistry, 134, 215–218. doi:10.1016/j.foodchem.2012.02.097. [45]
Navarro, R. D., Pinto, O., Filho, R., & Calado, L. L. (2006). Comparação morfometrica e índices somáticos de machos e fêmeas do lambari prata ( Astayanax scabripinnis Jerenyns, 1842) em diferente sistema de cultivo. Zootecnia Tropical, 24, 165–176. doi:S0798-72692006000200006.
[46]
Nelson, D. L., Lehninger, A. L., & Cox, M. M. (2008). Lehninger principles of biochemistry. Macmillan.
ACCEPTED MANUSCRIPT [47]
Neu, D., Furuya, W., Boscolo, W., Potrich, F., Lui, T., & Feiden, A. (2012). Glycerol inclusion in the diet of nile tilapia (oreochromis niloticus) juveniles. Aquaculture Nutrition, 19, 211––217. doi:10.1111/j.1365-2095.2012.00968.x.
[48]
NRC (2011). Nutrient Requirements of Fish and Shrimp. Washington, DC: The National Academies Press.
[49]
Ogawa, M., & Maia, E. L. (1999). Manual da Pesca Ciência e tecnologia do pescado. São
IP
[50]
T
Paulo: Varela.
Okamura, D., de Araújo, F. G., Rosa, P. V. e., de Freitas, R. T. F., Murgas, L. D. S., &
CR
Cesar, M. P. (2010). Influência da concentração de benzocaína e do comprimento dos peixes na anestesia e na recuperação de tilápias-do-nilo. Revista Brasileira de Zootecnia,
[51]
US
39, 971–976. doi:10.1590/S1516-35982010000500005. Omasaki, S., Janssen, K., Besson, M., & Komen, H. (2017). Economic values of growth Aquaculture, 481, 124–132. URL:
AN
rate, feed intake, feed conversion ratio, mortality and uniformity for nile tilapia.
M
https://doi.org/10.1016%2Fj.aquaculture.2017.04.013. doi:10.1016/j.aquaculture.2017.04.013. Peng, S., Chen, L., Qin, J. G., Hou, J., Yu, N., Long, Z., Ye, J., & Sun, X. (2008). Effects of
ED
[52]
replacement of dietary fish oil by soybean oil on growth performance and liver
PT
biochemical composition in juvenile black seabream, Acanthopagrus schlegeli. Aquaculture, 276, 154–161. doi:10.1016/j.aquaculture.2008.01.035. R Development Core Team (2013). R: A language and environment for statistical
CE
[53]
computing. R Foundation for Statistical Computing, Vienna, Austria. URL
[54]
AC
http://www.R-project.org/. Ribeiro, P. A. P., Logato, P. V. R., Paula, D. A. d. J., Costa, A. C., Murgas, L. D. S., & de Freitas, R. T. F. (2008). Efeito do uso de óleo na dieta sobre a lipogênese e o perfil lipídico de tilápias-do-nilo. Revista Brasileira de Zootecnia, 37, 1331–1337. doi:10.1590/S1516-35982008000800001. [55]
Sánches-Muros, M. J., García-Rejón, L., Lupiáñez, J. A., & Higuera, M. (1996). Long-term nutritional effects on the primary liver and kidney metabolism in rainbow trout (Oncorhynchus mykiss). II. Adaptive response of glucose 6-phosphate dehydrogenase activity to high-carbohydrate/low-protein and high-fat/non-carbohydrate diets.
ACCEPTED MANUSCRIPT Aquaculture Nutrition, 2, 193–200. doi:10.1111/j.1365-2095.1996.tb00059.x. [56]
Sousa, Á. B. B. d., Júnior, S., de Oliveira, O., Visentainer, J. V., & Almeida, N. M. d. (2017). Total lipid nutritional quality of the adipose tissue from the orbital cavity in nile tilapia from continental aquaculture. Acta Scientiarum. Animal Sciences, 39, 335–341.
[57]
Steffens, W. (1997). Effects of variation in essential fatty acids in fish feeds on nutritive
T
value of freshwater fish for humans. Aquaculture, 151, 97–119.
Vargas, R. J., Souza, S. M., Tognon, F., Gomes, M. E., & Kessler, A. d. M. (2007).
CR
[58]
IP
doi:10.1016/S0044-8486(96)01493-7.
Desempenho de alevinos de tilápias do nilo alimentados com dietas contendo diferentes
[59]
US
fontes de lipídeos. Revista Brasileira de Agrociência, 13, 377–381. Visentainer, J. V. (2012). Aspectos analíticos da resposta do detector de ionização em
AN
chama para ésteres de ácidos graxos em biodiesel e alimentos. Química Nova, 35, 274–279. doi:10.1590/s0100-40422012000200008. Wang, J.-T., Liu, Y.-J., Tian, L.-X., Mai, K.-S., Du, Z.-Y., Wang, Y., & Yang, H.-J.
M
[60]
(2005). Effect of dietary lipid level on growth performance, lipid deposition, hepatic
ED
lipogenesis in juvenile cobia (rachycentron canadum). Aquaculture, 249, 439–447. doi:10.1016/j.aquaculture.2005.04.038. Wood, J., Richardson, R., Nute, G., Fisher, A., Campo, M., Kasapidou, E., Sheard, P., &
PT
[61]
Enser, M. (2004). Effects of fatty acids on meat quality: a review. Meat Science, 66, 21–32.
[62]
CE
doi:10.1016/s0309-1740(03)00022-6. Xu, W.-n., Li, J.-y., Li, X.-f., Huang, G.-q., Liu, W.-b. et al. (2013). Alterations of liver
AC
histology and blood biochemistry in blunt snout bream megalobrama amblycephala fed high-fat diets. Fisheries Science, 79, 661–671. [63]
Yoneyama, Y., Yonemori, Y., Murata, M., Ohnuki, H., Hibi, K., Hayashi, T., Ren, H., & Endo, H. (2009). Wireless biosensor system for real-time cholesterol monitoring in fish “Nile tilapia”. Talanta, 80, 909–15. doi:10.1016/j.talanta.2009.08.014.
[64]
Zhou, P., Wang, M., Xie, F., Deng, D.-F., & Zhou, Q. (2016). Effects of dietary carbohydrate to lipid ratios on growth performance, digestive enzyme and hepatic carbohydrate metabolic enzyme activities of large yellow croaker (larmichthys crocea). Aquaculture, 452, 45–51. URL:
ACCEPTED MANUSCRIPT https://doi.org/10.1016%2Fj.aquaculture.2015.10.010.
AC
CE
PT
ED
M
AN
US
CR
IP
T
doi:10.1016/j.aquaculture.2015.10.010.
ACCEPTED MANUSCRIPT
AC
CE
PT
ED
M
AN
US
CR
IP
T
Soybean oil in increasing levels affect the fishes muscle composition; Vegetable oil has influence in fish nutrition; Soybean oil as energy and fat acids source; Soybean oil does increasing tilapias' development.