Importance of the proportions of dietary polyunsaturated fatty acids and antioxidants in larval

242 Pages·2008·3.06 MB·English

by  Atalah, Eyad

aAD DE LAS PALMAS DE GRAN CANARIA
Departamento de Biología
Grupode Investigación
en Acuicultura
Las Palmas de Gran Canaria, Spain
2008
DEPARTAMENTO DE BIOLOGIA
Universidad de Las Palmas de Gran Canaria
Anexo I
D. JUAN LUIS GÓMEZ PINCHETTI SECRETARIO DEL
DEPARTAMENTO DE BIOLOGíA DE LA UNIVERSIDAD DE LAS
PALMAS DE GRAN CANARIA,
CERTIFICA,
Que el Consejo de Doctores del Departamento en su sesión de
fecha hoy tomó el acuerdo de dar el consentimiento para su
tramitación, a la tesis doctoral titulada "IMPORTANCE OF THE
PROPORTIONS OF DIETARY POLYUNSATURATED FATTY
ACIDS ANO ANTIOXIDANTS IN LARVAL DEVELOPMENT OF
MARINE FISH" presentada por el doctorando D. EYAD ATALAH,
a
dirigida por la Doctora Da M SOLEDAD IZQUIERDO LÓPEZ y
a
codirigida por la Doctora Da CARMEN M HERNÁNDEZ CRUZ.
CAMPUS UNIVERSITARIO DE TAFIRA-35017Las Palmas-Islas Canarias-España-Tlf.: 34-92845 29 11 -Fax: [phone]
-
UNIVERSIDAD DE LAS PALMAS DE GRAN CANARIA
Departamento: Biologia & Instituto Universitario de Sanidad
animales (lUSA)
Programa de Doctorado: Acuicultura: producción controlada de
animales acuáticos
Titulo de la Tesis
Importance of the proportions of dietary
polyunsaturated fatty acids and antioxidants in larval
development of marine fish
Tesis Doctoral presentada por D. Eyad Atalah
Dirigida por la Catedrática. Marisol Izquierdo López
Codirigida por Prof. Carmen María Hernández Cruz
La Direct~ra La Codirectora El Doctorando
,1.
,
/
Las palmas de gran canaria, a 5 de Noviembre de 2008
aD CANARIA
DELASPAlMASDEGRAN
DepartamentodeBiología

Importance of the proportions of dietary
polyunsaturated fatty acids and antioxidants in larval
development of marine fish

EYAD ATALAH
Grupo de Investigación en Acuicultura
Instituto Canario de Ciencias Marinas
Universidad de Las Palmas de Gran
Canaria

Being a thesis submitted for the degree of
Doctor of Philosophy
In the University of Las Palmas de Gran Canaria, 2008
Directors:
Prof. Marisol Izquierdo & Prof. Carmen María Hernández Cruz
TABLE OF CONTENTS

List of tables ……………………………………………………….. III
List of figures ...…………………………………………………….. V
List of abbreviations ……………………………………………….. VIII
Acknowledgements ……………………………………………….. IX

1. Chapter 1: Introduction 1
1.1. Importance of fish lipid nutrition for aquaculture
development ………………………………………... 1
1.2. Essential fatty acids ………………………………… 2
1.3. Specific importance of DHA, EPA and ARA ……… 5
1.4. Importance of different ratios of DHA, EPA and
ARA ………………………………………………… 6
1.5. Importance of antioxidant vitamins in lipid nutrition . 8
1.6. Vitamin E …………………………………………… 9
1.7. Vitamin C …………………………………………... 11
1.8. Relation vitamin E and C …………………………... 12
1.9. Objectives……………………………………….…… 13
1.10. References…………………………………………… 18

2. Chapter 2: Materials and Methods 39
2.1. Fish …………………………………………………. 39
2.1.1. Gilthead sea bream ………………………… 39
2.1.2. Sea bass …………………………………….. 39
2.2. Experimental conditions ……………………………. 39
2.3. Diet and Feeding ……………………………………. 40
2.3.1. Diet formulation …………………………… 40
2.3.2. Microdiets preparation …………………….. 40
2.4. Larval feeding ……………………………………… 41
2.5. Sampling …………………………………………… 41
2.6. Activity test and survival …………………………... 42
2.7. Growth evaluations …………………………………. 42
2.8. Biochemical Analysis ………………………………. 42
2.8.1. Moisture …………………………………… 42
2.8.2. Protein ……………………………………... 43
2.8.3. Total Lipid ………………………………… 43
2.8.4. Separation of polar lipids…………………... 43
2.8.5. Fatty acid methyl esters preparation and
quantification ………………………………. 43
2.9. Statistical analysis …………………………………. 44
I
3. Chapter 3: Effect of different dietary EPA/DHA ratios on
incorporation of essential fatty acid in gilthead sea bream
larvae Sparus aurata (Linnaeus, 1758) ……………………… 49

4. Chapter 4: Importance of relative levels of dietary ARA and
EPA for culture performance of gilthead seabream (Sparus
aurata) larvae ……………………………………...………….. 71

5. Chapter 5: Importance of dietary arachidonic acid for survival,
growth and stress resistance of larval European sea bass fed
high dietary docosahexaenoic and eicosapentaenoic acids …… 89

6. Chapter 6: The effect of vitamin E and different DHA levels
on growth, survival, and stress resistance in sea bass
(Dicentrarchus labrax) larvae …………………………...……. 107

7. Chapter 7: Enhancement of gilthead seabream (Sparus aurata)
larval growth by dietary vitamin E in relation to two different
levels of essential fatty acids …………………………………. 127

8. Chapter 8: Combined effect of vitamin C and vitamin E
microdiets for gilthead sea bream Sparus aurata …………….… 149

9. Chapter 9: General Conclusions ………………………………. 171

10. Capítulo 10: Resumen amplio en Español ……………………. 173
10.1. Introducción ………………………………………... 174
10.2. Materiales y Métodos …………………………...…. 193
10.3. Resultados y discusión …………………………….. 203
10.4. Bibliografía………………………………………….. 207

II
LIST OF TABLES

Table 1.1 Optimal EPA/DHA or DHA/EPA ratio for different fish species …... 15
Table 1.2. Optimal vitamin E for different fish species ………………………... 16
Table 1.3. Optimal vitamin C for different fish species ………………………... 17
Table 2.1. Main fatty acids of the major dietary lipid sources used in these
experiments …………………………………………………………….. 45
Table 2.2. Mixture of attractants, minerals and vitamins ………………………. 46
Table 3.1. Lipid sources (% total ingredients) and crude lipid (% dry basis)
and moisture (% wet basis) content of the experimental diets ………… 52
Table 3.2. Main fatty acid composition of the experimental diets for gilthead
seabream fed microdiets containing different EPA/DHA proportions … 53
Table 3.3. Effect of dietary EPA and DHA levels on average survival and
resistance to handling stress of larval gilthead seabream ……………… 56
Table 3.4. Effect of dietary EPA and DHA levels on final total length and body
weight of gilthead sea bream …………………………………………... 56
Table 3.5. Average final lipid and fatty acid composition of larval gilthead
seabream ……………………………………………………………….. 61
Table 4.1. Lipid sources (% total ingredients), crude lipid (% dry basis), crude
protein (% dry basis) and moisture (% wet basis) content of the
experimental diets …………………..………………………………….. 74
Table 4.2. Fatty acid composition of experimental diets for larval gilthead
seabream ….…………………………………………………………….. 75
Table 4.3. Lipid content and fatty acid composition of total lipids from gilthead
seabream larvae after 14 days of feeding several EPA/ARA levels …… 82
Table 5.1. Main lipid ingredients composition and analyzed lipid and protein
contents of the experimental diets ……………………………………… 93
Table 5.2. Fatty acid composition of experimental diets for larval European sea
bass ……………………………………………………………………... 94
Table 5.3. Effect of dietary ARA levels on survival, growth and stress
resistance of European sea bass larvae …………………………………. 95

III
Table 5.4. Lipid content and fatty acid composition of total lipids of European
sea bass larvae fed several ARA levels ………………………………… 96
Table 6.1. Main lipid ingredients composition and vitamin E contents of the
experimental diets ……………….…………………………………… 110
Table 6.2. Fatty acid composition of the experimental diets for European sea
bass (g/100 g diet dry weight) ………………………………………….. 111
Table 6.3. Effect of different dietary DHA/vit E contents on larval growth, and
results of the two-way anova statistical analysis ……………….…..... 114
Table 6.4. Fatty acid composition of total lipid of the larval European sea bass
(% total fatty acid)..…………………………………………………...… 118
Table 7.1. Lipid sources (% total ingredients), α-tocopherol (mg/kg) and crude
lipid (% dry basis) content of the experimental diets …………………... 130
Table 7.2. Fatty acid composition of the experimental diets for gilthead
seabream (g/100 g diet dry weight) …………………………………….. 130
Table 7.3. Effect of different dietary DHA/EPA and vit E contents on larval
growth, and results of the two-way anova statistical analysis …………. 135
Table 7.4. Fatty acid composition of polar lipid of the larval gilthead seabream 139
Table 8.1. Dietary vitamin E and C composition ………………………………. 152
Table 8.2. Fatty acid composition of the experimental diets for gilthead
seabream ………………………………………………………………... 153
Table 8.3. Fatty acid composition of polar lipid of the larval gilthead seabream 159
Tabla 10.1. Relación Óptima de EPA/DHA, DHA/EPA, DHA/EPA/ARA y
ARA para las diferentes especies de peces …………………………… 189
Tabla 10.2. Óptimo de vitamina E para las diferentes especies de peces ……… 190
Tabla 10.3. Óptimo de vitamina C para las diferentes especies de peces ……… 191
Tabla 10.4. Principales ácidos grasos dietéticos de las principales fuentes
lípidicas utilizadas en los experimentos ………………………………... 200
Tabla 10.5. Mezcla de atractantes, minerales y vitaminas…………………….... 201

IV
LIST OF FIGURES

Figure 2.1. Tanks used for larval rearing during feeding experiments ………… 47
Figure 2.2. Larval microdiet formulated and prepared for the feeding
experiments…………………………………………………………… 47
Figure 3.1. Effect of dietary EPA (% d.w.) on survival of gilthead seabream
larvae fed different EPA/DHA ratio……………………………………. 55
Figure 3.2. Effect of dietary EPA and DHA (% d.w.) on survival of gilthead
seabream larvae fed different EPA/DHA ratio…………………………. 55
Figure 3.3. Effect of dietary n-3 HUFA (% d.w.) on average final total length
of gilthead seabream larvae fed different EPA/DHA ratio…………….. 57
Figure 3.4. Effect of (EPA+DHA+ARA)*(DHA/EPA)/ARA on total length of
gilthead seabream larvae fed several EPA/DHA ratio at the end of
trial1…………………………………………………………………….. 57
Figure. 3.5 Evolution of dietary oleic acid (% fatty acid) in diet and gilthead
seabream larvae (% fatty acid) fed different EPA/DHA ratio………….. 58
Figure 3.6. Evolution of EPA in diets and gilthead seabream larvae fed
different EPA/DHA ratio……………………………………………….. 58
Figure 3.7. Graphical representation of EPA (x axis) and DHA (y axis)
contents on diets fed to gilthead seabream and the larval growth (SGR,
secondary y)………………………………………………………….. 59
Figure 3.8. Graphical representation of EPA (x axis) and DHA (y axis)
contents on diets fed to gilthead seabream and the larval survival
(secondary y)……………………………………………………………. 60
Figure 4.1. Dietary ARA. EPA and DHA % dry basis………………………… 76
Figure 4.2. The survival rate of fish fed the different experimental diets………. 77
Figure 4.3. Dry whole body weight after 7 days of feeding……………………. 77
Figure 4.4. Final dry whole body weight of fish fed the different experimental
diets……………………………………………………………………... 78
Figure 4.5. Specific growth rate of fish fed the different experimental diets…... 78
Figure 4.6. Final standard length of fish fed the different experimental diets….. 79
Figure 4.7. Final biomass of fish fed the different experimental diets………… 80
V
Figure 4.8. Incorporation of dietary ARA in larvae tissues…………………….. 81
Figure 4.9. Incorporation of dietary EPA in larvae tissue………………………. 81
Figure 5.1. Evolution of body weight of fish fed the different experimental
diets…………………………………………………………………… 97
Figure 5.2. Evolution of standard length of fish fed the different experimental
diets…………………………………………………………………… 98
Figure 5.3. Effect of dietary ARA level on larval survival after handling stress
at the end of the experiment…………………………………………….. 98
Figure 5.4. Effect of dietary ARA level on larval survival……………………... 99
Figure 6.1. Survival rate of fish fed the experimental diets containing several
vit E and DHA………………………………………………………….. 113
Figure 6.2. Survival after 24h of air activity test of fish fed the different
experimental diets………………………………………………………. 114
Figure 6.3. Final total length of fish fed the different experimental diet……….. 115
Figure 6.4. Final dry whole body weight of fish fed the different experimental
diets…………………………………………………………………….. 115
Figure 6.5. Specific growth rate (SGR) of fish fed the different experimental
diets…………………………………………………………………… 116
Figure 6.6. Final biomass of fish fed the different experimental diets…………. 117
Figure 6.7. Correlation between dietary DHA levels (in the 1500 mg vit E
diets) and survival of fish fed the different experimental diets………… 120
Figure 7.1. Survival rate of fish fed the experimental diets containing several
vit E and DHA/EPA…………………………………………………….. 133
Figure 7.2. Correlation between the vitamin E levels (in the 5/2.5 diets) and
survival of fish fed the different experimental diets…………………… 134
Figure 7.3. Final total length of fish fed the different experimental diets……… 134
Figure 7.4. Final dry whole body weight of fish fed the different experimental
diets……………………………………………………………………. 135
Figure 7.5. Specific growth rate of fish fed the different experimental diets… 136
Figure 7.6. Final biomass of fish fed the different experimental diets………… 137
Figure 7.7. Survival after 24h of air activity test of fish fed the different
experimental diets..................................................................................... 137
Figure 7.8. Survival after 24h of exposure to a temperature reduction in fish
VI