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Lipid composition and fatty acid profiles of eggs from wild and cultured chinook salmon (oncorhynchus… Ashton, Heather 1991

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L I P I D COMPOSITION AND FATTY ACID PROFILES OF EGGS FROM WILD AND CULTURED CHINOOK SALMON (ONCORHYNCHUS TSHAWYTSCHA) BROODSTOCK by Heather B.Sc.  Ashton  U n i v e r s i t y o f V i c t o r i a , 1976.  A THESIS SUBMITTED I N P A R T I A L FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE STUDIES (Department o f Animal  Science)  We a c c e p t t h i s t h e s i s a s c o n f o r m i n g to the required standard  THE UNIVERSITY OF B R I T I S H COLUMBIA O c t o b e r 1991 ©  H e a t h e r A s h t o n , 1991  In  presenting this  thesis in partial  fulfilment  of the requirements  for an advanced  degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further copying  agree that permission for extensive  of this thesis for scholarly purposes may be granted  department  or  by  his or  her  representatives.  It  is  by the head of my  understood  that  copying or  publication of this thesis for financial gain shall not be allowed without my written permission.  Department of  M / A / M L  SCl£MC£  The University of British Columbia Vancouver, Canada  Date  DE-6 (2/88)  / /  o c r o 6 6 / e  is  91  ABSTRACT  In  Experiment  1, e g g s o f B i g Q u a l i c u m  (RC) w i l d and c u l t u r e d C h i n o o k tschawytcha) profiles.  (BQ)  and R o b e r t s o n  Creek  salmon (Oncorhynchus  were a n a l y z e d f o r l i p i d  c o m p o s i t i o n and  fatty  E a c h o f t h e c u l t u r e d b r o o d s t o c k s had b e e n f e d  acid  two  f o r m u l a t e d d i e t s d e s i g n a t e d a s COMM and WV33.  Significantly a c i d s and  h i g h e r c o n c e n t r a t i o n s o f s a t u r a t e d and  n3  l o w e r c o n c e n t r a t i o n s o f n6 and n9 f a t t y a c i d s w e r e  f o u n d i n b o t h t h e t o t a l and p o l a r  l i p i d s o f t h e eggs f r o m  w i l d f i s h than i n those from e i t h e r group of c u l t u r e d H i g h l y u n s a t u r a t e d f a t t y a c i d s (HUFAs), 22:6n3, were t h e major  The  l i p i d s w e r e up t o 5.3  i n t h e c u l t u r e d eggs.  primarily  the  fish.  20:5n3  and  c o n t r i b u t o r s t o t h e n3 s e r i e s i n b o t h  t h e w i l d and c u l t u r e d e g g s . and p o l a r  fatty  The  n3:n6 r a t i o s o f b o t h t h e times greater i n the w i l d  monounsaturate  total than  c o n c e n t r a t i o n was  s i g n i f i c a n t l y g r e a t e r i n t h e c u l t u r e d eggs t h a n i n t h e  wild  eggs.  The  f a t t y a c i d c o m p o s i t i o n o f t h e eggs g e n e r a l l y r e f l e c t e d  f a t t y a c i d p r o f i l e s of the d i e t s fed to the  broodstock.  C o n d i t i o n f a c t o r s f o r t h e w i l d f i s h were s i g n i f i c a n t l y than f o r e i t h e r group of c u l t u r e d f i s h , r e f l e c t i n g significantly  the  lower  the  g r e a t e r body l e n g t h o f t h e w i l d f i s h t h a n  the  cultured  fish.  T h e r e were no s i g n i f i c a n t d i f f e r e n c e s  weights.  Survival  t o e y e i n g among t h e c u l t u r e d  than that  reported  by t h e r e s p e c t i v e  hatcheries  The  for the wild  fertility, poorest fatty  incidences  i n both stocks  p a r t i c u l a r l y i n t h e RC f i s h .  i n t h e RC-WV33 e g g s .  responsible, generally COMM f i s h .  c l o s e r t o those of the wild T h e r e was no e x p l a n a t i o n  2,  feed  to transport  fish  acid profiles.  compromised by t h i s  for  t o e y e i n g was l i p i d and  than t o those o f the  for the i n f e r i o r  compared t o t h e BQ-WV33  fish.  was w i t h d r a w n f r o m a g r o u p o f C h i n o o k 7 day s t a r v a t i o n  t o freshwater f o r f i n a l  were no d i f f e r e n c e s  diet  Survival  ( p o s s i b i l y V i t a m i n C) a p p e a r e d t o be  b r o o d s t o c k f o r 7 d a y s beyond t h e u s u a l  Experiment  eggs a n d low  a s t h e f a t t y a c i d p r o f i l e s i n t h e WV33 eggs were  In e x p e r i m e n t  fatty  f e d t h e WV33 d i e t ,  Some f a c t o r o t h e r t h a n  p e r f o r m a n c e o f t h e RC-WV33 f i s h  prior  government  o f a b n o r m a l and r e t a i n e d  a c i d composition  eggs was l o w e r  eggs.  q u a l i t y o f eggs was p o o r  with high  federal  i n body  in lipid  maturation.  period There  c o m p o s i t i o n and few d i f f e r e n c e s i n  F e c u n d i t y a n d s u r v i v a l t o e y e i n g were n o t treatment.  3 was c a r r i e d o u t t o d e t e r m i n e w h e t h e r a f o r m u l a t e d  (COMM) augmented w i t h k r i l l  (designated  s i x weeks p r i o r t o t h e p r e - t r a n s p o r t  a s t h e BROOD d i e t )  starvation  period  iv  would a f f e c t l i p i d  composition,  incubation success.  f a t t y a c i d p r o f i l e s and  The c o n t r o l d i e t was t h e COMM f o r m u l a t i o n .  The BROOD e g g s were s i g n i f i c a n t l y l o w e r i n t o t a l concentration lipid  due t o t h e p r e s e n c e o f s i g n i f i c a n t l y l e s s n e u t r a l  t h a n i n t h e COMM e g g s .  individual  lipid  Small  significant  differences i n  f a t t y a c i d s were f o u n d b e t w e e n t h e t w o g r o u p s .  W h i l e some o f t h e s e d i f f e r e n c e s r e f l e c t e d t h e d i e t a r y acids (ie. acids  (eg. 2 2 : l n l l ) ,  others  a p p e a r e d t o be due t o r e t e n t i o n  n3 f a t t y a c i d s ) o r t o e l o n g a t i o n (eg. 18:ln9).  and d e s a t u r a t i o n o f f a t t y  No d i f f e r e n c e s i n i n c u b a t i o n  found between t h e groups.  fatty  success  were  V  TABLE OF CONTENTS Page ABSTRACT  i i  TABLE OF CONTENTS L I S T OF FIGURES L I S T OF TABLES ACKNOWLEDGEMENTS  v viii ix xiv  INTRODUCTION  1  LITERATURE REVIEW  7  1 2 3 4 5 6 7 8  L i p i d s o f p h y s i o l o g i c a l importance f o r f i s h D i e t a r y f a t t y a c i d s and r e q u i r e m e n t s f o r g r o w t h Digestion of dietary l i p i d s Role of l i p i d s i n f i s h F a t t y a c i d s y n t h e s i s and m o b i l i z a t i o n Vitellogenesis Embryonic development Feeding regime f o r c u l t u r e d maturing salmonids  7 9 13 15 23 28 32 34  SECTION 1 - A c o m p a r i s o n o f t h e l i p i d c o m p o s i t i o n and f a t t y a c i d p r o f i l e s o f t h e eggs o f two s t o c k s o f w i l d and c u l t u r e d C h i n o o k s a l m o n . 1.1  M a t e r i a l s and Methods 1.1.1 E x p e r i m e n t a l d e s i g n and c o n d i t i o n s 1.1.2 Sample c o l l e c t i o n 1.1.3 Egg c o m p o s i t i o n 1.1.4 Diet composition 1.1.5 Egg s i z e 1.1.6 F e r t i l i z a t i o n and e y e i n g s u c c e s s 1.1.7 S t a t i s t i c a l procedures  37 37 41 42 49 50 51 51  1.2  R e s u l t s and D i s c u s s i o n 1.2.1 M o r p h o m e t r i c measurements 1.2.2 Spawning and i n c u b a t i o n s u c c e s s 1.2.3 C o m p o s i t i o n o f t h e eggs 1.2.4 Fatty acid composition of the t o t a l l i p i d s 1.2.5 Fatty acid composition of the polar l i p i d s 1.2.6 Diets  53 53 55 63 68 81 93  1.3  Conclusions  101  vi SECTION 2 - F e e d w i t h d r a w a l f r o m c u l t u r e d C h i n o o k b r o o d s t o c k p r i o r to t r a n s f e r to freshwater f o r maturation. 2.1  M a t e r i a l s and Methods  109  2.2  R e s u l t s and D i s c u s s i o n 2.2.1 M o r p h o m e t r i c measurements 2.2.2 Spawning and i n c u b a t i o n s u c c e s s 2.2.3 C o m p o s i t i o n o f t h e eggs 2.2.4 Fatty a c i d composition of the t o t a l l i p i d s 2.2.5 Fatty a c i d composition of the polar l i p i d s  111 111 111 115 118 123  2.3  Conclusions  130  SECTION 3 - A l t e r a t i o n o f l i p i d c o m p o s i t i o n and f a t t y a c i d p r o f i l e s i n eggs f r o m C h i n o o k b r o o d s t o c k . 3.1  M a t e r i a l s and Methods  131  3.2  R e s u l t s and D i s c u s s i o n 3.2.1 M o r p h o m e t r i c measurements 3.2.2 Spawning and i n c u b a t i o n s u c c e s s 3.2.3 C o m p o s i t i o n o f t h e eggs 3.2.4 Fatty acid composition of the t o t a l l i p i d s 3.2.5 Fatty acid composition of the polar l i p i d s 3.2.6 Diets  133 133 133 136 138 140 147  3.3  Conclusions  152  CONCLUDING REMARKS  153  REFERENCES  155  APPENDIX 1 - P r e l i m i n a r y a n a l y s e s o f s e l e c t e d n u t r i e n t p a r a m e t e r s i n t h e eggs o f w i l d and c u l t u r e d C h i n o o k salmon - 1 9 8 5 & 1 9 8 6 .  164  APPENDIX 2 - D i e t  Data.  Table A 2 . 1 : Manufacturer's feed l a b e l data f o r c o m m e r c i a l (COMM) and b r o o d (BROOD) diets. T a b l e A 2 . 2 : F o r m u l a t i o n o f t h e West V a n c o u v e r 3 3 (WV33)  diet.  165 166  T a b l e A 2 . 3 : Dry m a t t e r and l i p i d c o m p o s i t i o n o f t h e e x p e r i m e n t a l d i e t s used i n S e c t i o n s 1 , 2 and 3 . 167 Table A 2 . 4 : Fatty a c i d p r o f i l e s of d i e t s . 168 T a b l e A2.5: Dry m a t t e r and l i p i d c o m p o s i t i o n and f a t t y acid p r o f i l e of a euphausiid (krill) sample. 169  vii  APPENDIX 3 - F o r m u l a e f o r c a l c u l a t i n g parameters. APPENDIX 4 - F a t t y a c i d Table A 4 . 1 :  Table A 4 . 2 :  Table A 4 . 3 :  Table A 4 . 4 :  Table A 4 . 5 :  T a b l e A4.6  Table A 4 . 7  Table A 4 . 8 :  composition 170  p r o f i l e s o f eggs.  F a t t y a c i d p r o f i l e s o f TOTAL LIPIDS i n t h e eggs o f BIG QUALICUM b r o o d s t o c k on t h r e e diets. 171 F a t t y a c i d p r o f i l e s o f TOTAL L I P I D S i n t h e eggs o f ROBERTSON CREEK b r o o d s t o c k on three d i e t s . 172 F a t t y a c i d p r o f i l e s o f POLAR LIPIDS i n t h e eggs o f BIG QUALICUM b r o o d s t o c k on t h r e e diets. 173 F a t t y a c i d p r o f i l e s o f POLAR L I P I D S i n t h e eggs o f ROBERTSON CREEK b r o o d s t o c k on three d i e t s . 174 F a t t y a c i d p r o f i l e s o f TOTAL LIPIDS i n t h e eggs o f 4 - y e a r o l d BIG QUALICUM b r o o d s t o c k . Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 days o r 1 4 days p r i o r t o t r a n s f e r t o freshwater f o r maturation. 175 F a t t y a c i d p r o f i l e s o f POLAR LIPIDS i n t h e eggs o f 4 - y e a r o l d BIG QUALICUM b r o o d s t o c k . F e e d was w i t h d r a w n f r o m t h e s e f i s h f o r 7 days o r 1 4 days p r i o r t o t r a n s f e r t o freshwater f o r maturation. 176 F a t t y a c i d p r o f i l e s o f TOTAL LIPIDS i n t h e eggs o f 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l grower d i e t (COMM) o r on a b r o o d (BROOD) d i e t p r i o r t o t r a n s f e r t o freshwater f o r maturation. 177 F a t t y a c i d p r o f i l e s o f POLAR LIPIDS i n t h e eggs o f 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l grower d i e t (COMM) o r on a b r o o d (BROOD) d i e t p r i o r t o t r a n s f e r t o freshwater f o r maturation. 178  viii L I S T OF FIGURES Figure 1  Page Conversion of arachidonic eicosanoids. Fatty  3  Flowchart of procedures f o r l i p i d fatty acid determinations.  5  6  7  8  synthesis  (20:4n6) t o  2  4  acid  acid  Egg a b n o r m a l i t i e s WV33 d i e t .  20  infish.  25 a n a l y s i s and 43  i n C h i n o o k b r o o d s t o c k on t h e 60  A c o m p a r i s o n o f s a t u r a t e d , n3, n6 a n d n9 f a t t y a c i d s a n d n3:n6 r a t i o s i n t o t a l a n d p o l a r l i p i d s o f eggs f r o m c u l t u r e d a n d w i l d B i g Q u a l i c u m a n d R o b e r t s o n C r e e k b r o o d s t o c k w i t h t h e COMM a n d WV33 diets.  95  A c o m p a r i s o n o f s e l e c t e d n3 f a t t y a c i d s , n3 PUFAs and HUFAs i n t o t a l and p o l a r l i p i d s o f eggs f r o m c u l t u r e d and w i l d B i g Qualicum and R o b e r t s o n Creek b r o o d s t o c k w i t h t h e COMM and WV33 d i e t s .  97  A c o m p a r i s o n o f s e l e c t e d f a t t y a c i d s a n d n6 PUFAs i n t o t a l a n d p o l a r l i p i d s o f eggs f r o m c u l t u r e d and w i l d B i g Q u a l i c u m a n d R o b e r t s o n C r e e k b r o o d s t o c k w i t h t h e COMM a n d WV33 d i e t s .  98  A comparison o f s e l e c t e d f a t t y a c i d s i n t h e t o t a l and p o l a r l i p i d s o f eggs o f 4-year o l d b r o o d s t o c k w i t h t h o s e i n t h e i r d i e t s (COMM a n d BROOD).  150  \  ix L I S T OF TABLES Table  1  5a 5b 6a  6b  7a  7b  8a  8b  9a  Page M o r p h o m e t r i c d a t a f o r w i l d and c u l t u r e d B i g Q u a l i c u m and R o b e r t s o n C r e e k f e m a l e b r o o d s t o c k .  54  Spawning d a t a f o r w i l d and c u l t u r e d and R o b e r t s o n C r e e k b r o o d s t o c k .  56  B i g Qualicum  Egg r e t e n t i o n and o t h e r r e p r o d u c t i v e a b n o r m a l i t i e s o b s e r v e d i n B i g Q u a l i c u m and R o b e r t s o n C r e e k c u l t u r e d f e m a l e s on two d i e t s .  58  I n c u b a t i o n d a t a f o r w i l d and c u l t u r e d and R o b e r t s o n C r e e k b r o o d s t o c k .  62  B i g Qualicum  C o m p o s i t i o n o f t h e eggs o f BIG QUALICUM on t h r e e d i e t s .  broodstock 64  C o m p o s i t i o n o f t h e eggs o f ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s .  65  S a t u r a t e d , n 3 , n6 a n d n9 f a t t y a c i d s e r i e s and n 3 : n 6 r a t i o s i n TOTAL LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e d i e t s .  70  S a t u r a t e d , n 3 , n6 and n9 f a t t y a c i d s e r i e s and n 3 : n 6 r a t i o s i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s .  71  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n TOTAL L I P I D S o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e d i e t s .  73  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs a n d t o t a l n3 HUFAs i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s .  74  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n TOTAL L I P I D S o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e d i e t s .  76  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n TOTAL L I P I D S o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on three d i e t s .  77  M o n o u n s a t u r a t e s , r e p o r t e d a s n 5 , n 7 , n 9 , n i l and t o t a l monounsaturated f a t t y a c i d s i n TOTAL LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e diets.  79  X  9b  10a  10b  11a  lib  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, r i l l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e diets.  80  S a t u r a t e d , n3, n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e d i e t s .  82  S a t u r a t e d , n3, n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s i n POLAR LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s .  83  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e d i e t s .  85  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n POLAR LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s .  86  12a  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e diets. 88  12b  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n POLAR LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on three d i e t s .  89  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM b r o o d s t o c k on t h r e e diets.  91  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n POLAR LIPIDS o f eggs f r o m ROBERTSON CREEK b r o o d s t o c k on t h r e e diets.  92  13a  13b  14  15  16  Morphometric d a t a f o r B i g Qualicum 4-year o l d broodstock. Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 days p r i o r t o f r e s h w a t e r t r a n s f e r .  112  Spawning and i n c u b a t i o n d a t a f o r B i g Q u a l i c u m 4-year o l d b r o o d s t o c k . Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 days p r i o r t o f r e s h w a t e r transfer.  113  C o m p o s i t i o n o f t h e eggs o f 4-year o l d B i g Q u a l i c u m broodstock. Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 days p r i o r t o f r e s h w a t e r t r a n s f e r .  116  xi  17  A b s o l u t e volumes o f t o t a l and p o l a r l i p i d p e r egg c a l c u l a t e d f r o m mean egg volumes and p e r c e n t a g e s o f these parameters. 117  18  S a t u r a t e d , n3, n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 days p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  119  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r to t r a n s f e r to freshwater f o r maturation.  120  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r to freshwater f o r maturation.  121  19  20  21  M o n o u n s a t u r a t e s , r e p o r t e d as n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o f r e s h w a t e r for maturation. 122  22  A b s o l u t e volumes o f t o t a l s a t u r a t e s , n3 f a t t y a c i d s , n3 HUFAs and 22:6n3 i n t h e t o t a l and p o l a r l i p i d s p e r egg c a l c u l a t e d f r o m t h e a b s o l u t e v o l u m e s o f t o t a l and p o l a r l i p i d p e r egg and t h e p e r c e n t a g e o f t h e s e p a r a m e t e r s , as r e p o r t e d i n p r e v i o u s t a b l e s . 124  23  S a t u r a t e d , n3, n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 days p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  126  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r to t r a n s f e r to freshwater f o r maturation.  127  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r to freshwater f o r maturation.  128  24  25  xii 26  M o n o u n s a t u r a t e s , r e p o r t e d a s n 5 , n 7 , n 9 , n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n POLAR LIPIDS o f eggs f r o m 4 - y e a r B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o f r e s h w a t e r for maturation. 129  27  Morphometric data f o r 4-year o l d B i g Qualicum b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o r maturation.  134  Spawning and i n c u b a t i o n d a t a f o r B i g Q u a l i c u m 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l d i e t (COMM) o r a b r o o d d i e t (BROOD) p r i o r t o t r a n s f e r t o freshwater f o r maturation.  135  C o m p o s i t i o n o f t h e eggs o f 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l (COMM) d i e t p r i o r t o t r a n s f e r t o freshwater f o r maturation.  137  28  29  30  S a t u r a t e d , n 3 , n6 and n9 f a t t y a c i d s e r i e s and n 3 : n 6 r a t i o s i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o transfer to freshwater f o r maturation. 139  31  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs a n d t o t a l n3 HUFAs i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o transfer to freshwater f o r maturation.  141  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o r maturation.  142  Monounsaturates, r e p o r t e d as n 5 , n 7 , n 9 , n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n TOTAL LIPIDS on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  143  32  33  34  S a t u r a t e d , n 3 , n6 and n9 f a t t y a c i d s e r i e s a n d n 3 : n 6 r a t i o s i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o transfer to freshwater f o r maturation. 144  xiii  35  36  37  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs and t o t a l n3 HUFAs i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o transfer to freshwater f o r maturation.  145  S e l e c t e d n6 f a t t y a c i d s and t o t a l n6 PUFAs i n POLAR LIPIDS o f eggs f r o m 4-year o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o r maturation.  146  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s i n POLAR LIPIDS o f eggs f r o m 4 - y e a r B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  148  xiv  ACKNOWLEDGEMENTS My s i n c e r e a p p r e c i a t i o n goes t o a l l who p r o v i d e d s u p p o r t and e n c o u r a g e m e n t t o me t h r o u g h o u t my s t u d i e s . In p a r t i c u l a r I w i s h t o t h a n k Dr. D a v i d G r o v e s f o r so w i l l i n g l y a c c e p t i n g me as a s t u d e n t a t Sea S p r i n g Salmon Farm and f o r h i s g e n e r o u s s h a r i n g o f knowledge and p r a c t i c a l e x p e r i e n c e i n t h e c u l t u r e and n u t r i t i o n o f s a l m o n . I w i l l never f o r g e t h i s u n f a i l i n g good humour and r e a d y t a l e s o f h i s y e a r s as a s t u d e n t and university professor. I w i s h a l s o t o t h a n k Dr. B e r y l March f o r h e r g u i d a n c e and knowledge and f o r b r i n g i n g t o my a t t e n t i o n t h e g r a m m a t i c a l faux pas o f t h e s p l i t i n f i n i t i v e . Her q u e s t i o n i n g n a t u r e , f a s c i n a t i o n w i t h t h e b i o l o g y o f a l l a n i m a l s and i n s i g h t s i n t o n u t r i t i o n were an i n s p i r a t i o n . Thank you t o Dr. D a v i d H i g g s who p r o v i d e d u s e f u l s u g g e s t i o n s and e n c o u r a g e m e n t t h r o u g h o u t . H i s p o s i t i v e a t t i t u d e toward r e s e a r c h p r o b l e m s and l i f e i n g e n e r a l and h i s sunny d i s p o s i t i o n were g r e a t l y a p p r e c i a t e d . T h a n k s a l s o t o Doug Harpham and t h e s t a f f ( e s p e c i a l l y P e t e r and C a t h y ) o f Sea S p r i n g Salmon Farm L t d ; G i l l e s G a l z y , Andy Hickman and L e s l i e H a r t o f t h e D e p a r t m e n t o f A n i m a l S c i e n c e , UBC; G o r d o n M i l l e r , S u s a n K e l l e r , H e n r i k K r i e b e r g and I a i n Whyt o f t h e P a c i f i c B i o l o g i c a l S t a t i o n i n Nanaimo and West V a n c o u v e r Yvonne Y o l e o f t h e S c a l e A g e i n g Lab a t DFO, V a n c o u v e r ; and Don L a w s e t h and G r a n t L a d o c o e u r , managers o f R o b e r t s o n C r e e k and B i Qualicum H a t c h e r i e s . T h a n k s a l s o t o my f a m i l y - E r i c , M e r r e l l , Mum and Dad, L a u r i e , B r i a n , J o h n , Elma and M a r g a r e t f o r t h e i r l o v e , p a t i e n c e and e n c o u r a g e m e n t t h r o u g h o u t my y e a r s as a s t u d e n t . T h i s work was C h e m a i n u s , BC award.  s u p p o r t e d by Sea S p r i n g Salmon Farm L t d . , and by t h e BC S c i e n c e C o u n c i l t h r o u g h a GREAT  F i n a l l y I w o u l d l i k e t o d e d i c a t e t h i s t h e s i s t o my 17 month o l d d a u g h t e r , M e r r e l l , who w i l l ' l e a r n much more t h a n I ' l l e v e r know.' 1  "I h e a r b a b i e s c r y ; I w a t c h them grow, T h e y ' l l l e a r n much more t h a n , I ' 1 1 e v e r know, And I t h i n k t o m y s e l f : What a W o n d e r f u l W o r l d . " L o u i s Armstrong,  "What a W o n d e r f u l  World."  1 INTRODUCTION  The  s u r v i v a l of c u l t u r e d  tschawytcha) has,  from f e r t i l i z a t i o n  until  This the of  natal  reported  f i g u r e was  by  to  Chinook,  35%.  release  eyeing  of  90%  as  Spring  the  salmon f a r m i n g with the  F i s h e r i e s and  Island  wild.  and  Survival  best  using  s u r v i v a l to  approximately  industry had  and  70%.  s u r v i v a l to  been i n t h e  been e v e n  lower.  Oceans, B i g  held  range Wild  system  and  85%  to (D.  Qualicum  F i s h e r i e s and  to  rear  the  Oceans f o r  60%  under i d e n t i c a l  private hatcheries  for  twelve  Chemainus  smolts f o r  to eyeing to  smolts  government  Chinook from the  rates  have been 20  rears  a federal  eggs f r o m c u l t u r e d B i g  reared the  has  spawn m a t u r i n g  rates of  of  the  a company t h a t  Department of  Chemainus f i s h  inability  incubated  to ponding of approximately  industry,  Vancouver  spawned and  Chinook  r o u t i n e l y have s u r v i v a l r a t e s  Salmon Farm L t d . ,  y e a r s t o t a k e and  survival  f o r the  ponding  comm.).  Sea  back t o t h e  of w i l d  f e d e r a l government h a t c h e r y  o r more and  Hatchery, pers.  wild  high  smolts,  Lawseth, Department of  on  1987,  some p r i v a t e h a t c h e r i e s  spawned i n t h e  to  River  Until  S u r v i v a l t o p o n d i n g had  reared  contract  through to  spawned and  a p r i v a t e h a t c h e r y was  unusually  eyed stage a t 13  t o h a t c h and  s t r e a m s and  same h a t c h e r y p r o c e d u r e s .  ponding  (Oncorhynchus  r e c e n t l y , b e e n much l o w e r t h a n t h a t  t a k e n from t h e i r the  C h i n o o k salmon  release  f o r eggs f r o m  better  than  Qualicum  conditions.  In  the  the  broodstock, 1986,  t o p r o d u c e enough  the  eggs  2 c o n s i s t e n t l y f o r t h e growing salmon f a r m i n g i n d u s t r y i n B r i t i s h Columbia c a l l e d  into question the v i a b i l i t y  o f salmon f a r m i n g  as a whole.  Poor s u r v i v a l  r a t e s a l s o hampered t h e d e v e l o p m e n t  s t r a i n s of f i s h  of domestic  selected f o r tolerance to stress,  r e s i s t a n c e , enhanced  growth r a t e s , improved  disease  reproductive  performance, e t c . w h i l e a v o i d i n g the negative impacts of inbreeding. could i l l  I n 1985  W i t h o u t a d e q u a t e numbers o f p r o g e n y ,  hatcheries  a f f o r d to undertake genetic experiments.  a n d 1986, p r e l i m i n a r y i n v e s t i g a t i o n s i n t o t h e p r o b l e m  o f p o o r egg s u r v i v a l  i n c u l t u r e d Chinook salmon were  by D r . D a v i d G r o v e s  (Sea S p r i n g S a l m o n Farm L t d . ) i n  c o n j u n c t i o n w i t h t h e B r i t i s h C o l u m b i a Salmon  Farmers  A s s o c i a t i o n and t h e G o v e r n m e n t o f C a n a d a , D e p a r t m e n t F i s h e r i e s and O c e a n s (IRAP  initiated  of  ( P I L P ) #CA910-5-0042/866).  B r o o d s t o c k h a n d l i n g and s p a w n i n g t e c h n i q u e s w e r e e x a m i n e d t h e major emphasis  o f t h e i n v e s t i g a t i o n was  broodstock n u t r i t i o n .  directed  With the p a r t i c i p a t i o n of  but  to  several  p r i v a t e h a t c h e r i e s , a survey of s e l e c t e d n u t r i e n t parameters i n c u l t u r e d e g g s and i n t h e e g g s f r o m t h r e e w i l d c a r r i e d out.  R e s u l t s f r o m t h e 1985  presented i n Appendix  was  analyses,  1, r e v e a l e d i m p o r t a n t d i f f e r e n c e s i n  f a t t y a c i d c o m p o s i t i o n between  The  and 1986  stocks  c u l t u r e d and w i l d  eggs.  o b j e c t i v e s o f t h i s t h e s i s were s e t o u t i n v i e w o f t h e s e  3 results. lipid  The  first  profiles  g o a l was  the accurate determination  i n t h e e g g s o f w i l d and  p u r p o s e , t h e e g g s o f two  stocks  cultured fish.  ( B i g Q u a l i c u m and  Creek) o f Chinook salmon were sampled.  Two  i n the  diet  (D.A.  Higgs,  B r i t i s h Columbia.  unpub. d a t a ) .  documenting l i p i d  These d i e t s  and  polar lipids  first  time, are presented  The  r e s u l t s of t h i s  i n Section  t h a n w i l d f i s h and  unpub. r e p o r t ) , i t was  (Appendix 1 ) .  levels  A  of  general  I t was  postulated  n o t t e n d t o be p r o d u c t i v e b r o o d s t o c k '  as p r o t e i n .  in their daily  lives,  and  being deposited  Reasons proposed f o r t h i s  t h a t c u l t u r e d f i s h expend l e s s energy than counterparts  higher  a l s o made t h a t c o n d i t i o n f a c t o r s w e r e l o w e r  i n the c u l t u r e d broodstock.  r a t h e r than  the  1.  t h a t i n g e s t e d e n e r g y i n c u l t u r e d f i s h was fat  the  t h a t c u l t u r e d f i s h p r o d u c e d eggs w i t h  concentrations  ' f a t f i s h do  two  study,  f a t t y a c i d p r o f i l e s of  f o u n d t h a t t h e m u s c l e o f c u l t u r e d f i s h had  the w i l d than  the  i n t h e eggs of Chinook salmon f o r  t h e e a r l y s t u d i e s (D. G r o v e s , 1987  o b s e r v a t i o n was  were  Robertson Creek H a t c h e r i e s ,  c o n c e n t r a t i o n s and  total  lipid  33  O c e a n s , West  W i l d C h i n o o k eggs were sampled f r o m  G o v e r n m e n t o f Canada f a c i l i t i e s .  that  i n wide  independent commercial f e e d companies i n  r e t u r n s t o t h e B i g Q u a l i c u m and  higher  this  groups of c u l t u r e d  D e p a r t m e n t o f F i s h e r i e s and  m a n u f a c t u r e d by two  lipid  For  i n d u s t r y o r t h e o p e n f o r m u l a t i o n , West V a n c o u v e r  Vancouver Laboratory,  In  the  Robertson  f i s h o f each s t o c k were f e d e i t h e r a c o m m e r c i a l d i e t use  of  their  as  were  wild  t h a t t h e y a r e on a  higher  in  4 plane  of n u t r i t i o n  than w i l d f i s h  i m b a l a n c e may e x i s t  or that  some  nutritional  which promotes t h e d e p o s i t i o n o f f a t r a t h e r  than p r o t e i n .  Wild  Chinook r e t u r n i n g t o t h e i r  cease feeding stocks  upon o r p r i o r  then undergo  spawning g r o u n d s .  the  extensive  Greene  prior  ( 1 9 1 9 ) and o t h e r s  d e p l e t i o n o f body r e s e r v e s  This  lends  fish  a r e on a h i g h e r  that  this  higher  production  test  fish  to freshwater.  Many  to reaching  have  reported  i n migrating  Chinook  t h e p e r i o d when gonad t i s s u e s a r e p r o l i f e r a t i n g and  maturing.  To  to entry  long arduous m i g r a t i o n s  the  during  n a t a l streams from t h e ocean  credence t o the suggestion plane  feeding  of high  of n u t r i t i o n  than  that cultured  i s necessary  r e g i m e may be d e t r i m e n t a l  to the  q u a l i t y eggs.  the hypothesis  that the plane  of n u t r i t i o n  i n cultured  t o w a r d s t h e end o f v i t e l l o g e n e s i s i s e x c e s s i v e ,  withdrawn from a group o f c u l t u r e d f i s h transfer  t o freshwater  f o r maturation  g r o u p , t h e c o n t r o l , was s t a r v e d following  the usual  composition  and  protocol  and s u r v i v a l  feed  was  f o r 14 d a y s p r i o r t o  and s p a w n i n g .  f o r one week p r i o r  A second to transfer,  f o r broodstock transport.  Lipid  r a t e s o f t h e eggs o f b o t h g r o u p s were  monitored.  The  third  profile  o b j e c t i v e was aimed a t m o d i f y i n g  the fatty  acid  o f t h e eggs o f c u l t u r e d b r o o d f i s h by f e e d i n g a  commercially  manufactured brood d i e t  containing  krill  for six  5 weeks p r i o r  to t r a n s p o r t to the  enrichment with of  fatty  fish  krill  provided  by  Sea  diet  and  a l l cultured fish  Robertson  Canada, D e p a r t m e n t o f  Creek f i s h  and  four other Pacific  the  study  BC  The  L t d . a t Genoa Bay sites.  from the diet  Fisheries  and  years  n e t pen  Fish  al.,  o f age  from  i n Section  by  and  1986).  when t h e  33  t h e Government  Fish;  of  of  BC This  study  f o r r e a r i n g these  and  diverse sites  along  o f Sea  Salmon Farm  Spring  V a n c o u v e r I s l a n d was  from each s t o c k ,  3.  The  Oceans; t h e Government  facility  n e a r Duncan on  same g e n e t i c mix  et  i n eggs  Salmon F a r m e r s ' A s s o c i a t i o n .  the  one  r e a r e d a t each s i t e ,  of were  a l l were f e d t h e West V a n c o u v e r  t o e l i m i n a t e g e n e t i c and  (Withler  balance  f o r t h i s work were  conducted  stocks at environmentally  coast.  that  f e d t h e West V a n c o u v e r  u n d e r t a k e n t o examine t h e p o t e n t i a l  these  found  C o l u m b i a , M i n i s t r y o f A g r i c u l t u r e and  Research;  33  be  T h i s work i s p r e s e n t e d  were p a r t o f a l a r g e r  British  thought  S p r i n g Salmon Farm L t d . , Chemainus, BC.  B i g Q u a l i c u m and  was  t o what may  f e e d i n g i n the w i l d .  facilities  I t was  w o u l d p r o v i d e a more f a v o u r a b l e  acids, i e . closer  Hatchery  hatchery.  diet  Some f i s h  sampling  factors  matured  from the  i n 1987  f o r the present  at  study  study three  was  undertaken. y  S i n c e t h e p r e l i m i n a r y s t u d i e s i n 1985 unpub. r e p o r t ) ,  the commercial  modifications to t h e i r higher  levels  C and  1986  (D. G r o v e s ,  1987  f e e d c o m p a n i e s have made  formulations  of Vitamins  and  including  E and  the  selenium,  a d d i t i o n of  demonstrated  as  6 essential  f o r the reproductive  (Watanabe, 1985). and b e c a u s e  Because  s u c c e s s o f many  the d i e t s are closed  species formulations  no c o n t r o l l e d e x p e r i m e n t s were p e r f o r m e d t o t e s t  their effects,  i t i s impossible t o a s c e r t a i n p r e c i s e l y which  m o d i f i c a t i o n s have been r e s p o n s i b l e f o r t h e  significant  i n c r e a s e i n s u r v i v a l o f c u l t u r e d e g g s s e e n s i n c e 1986. perhaps not as c o n s i s t e n t l y s u c c e s s f u l as t h e i r w i l d counterparts, survival  c u l t u r e d Chinook  s a l m o n t y p i c a l l y now  r a t e s t o p o n d i n g o f 75-85%.  have  While  7  LITERATURE REVIEW  1  L i p i d s of p h y s i o l o g i c a l importance f o r f i s h  L i p i d s i n c l u d e f a t s , o i l s , waxes a n d r e l a t e d compounds. are  relatively  They  i n s o l u b l e i n water but s o l u b l e i n nonpolar  s o l v e n t s such as c h l o r o f o r m ,  ether  and benzene.  Fatty acids are constituents of a l l l i p i d s . hydrocarbon chain with a terminal carboxylate  They c o n s i s t o f a group.  Fatty  a c i d s c a n be d i v i d e d i n t o two b r o a d c a t e g o r i e s , t h e s a t u r a t e s and t h e u n s a t u r a t e s . bonds.  Saturated  f a t t y a c i d s h a v e no d o u b l e  U n s a t u r a t e d f a t t y a c i d s h a v e d o u b l e b o n d s b e t w e e n some  o f t h e c a r b o n atoms i n t h e h y d r o c a r b o n c h a i n a n d may monoenoic, d i e n o i c , t r i e n o i c ,  called  ... h e x a e n o i c i f o n e , t w o ,  ... s i x d o u b l e bonds a r e p r e s e n t .  three,  C o n s e c u t i v e d o u b l e bonds a r e  m e t h y l e n e i n t e r r u p t e d so t h a t a -CH 2  carbons c a r r y i n g double bonds.  be  g r o u p o c c u r s b e t w e e n two  The p o s i t i o n o f t h e f i r s t  d o u b l e b o n d f r o m t h e m e t h y l end o f t h e m o l e c u l e i n  unsaturates  i s o f p h y s i o l o g i c a l s i g n i f i c a n c e a n d d e t e r m i n e s t h e omega (n) number o f t h e f a t t y a c i d [ 1 1 . The f a t s a n d o i l s  are u s u a l l y considered  i n s o l u b l e i n water  due  [1] F a t t y a c i d n o m e n c l a t u r e f o l l o w s t h e n o t a t i o n C r x n p , where C = t h e number o f c a r b o n atoms i n t h e h y d r o c a r b o n c h a i n , x = t h e number o f d o u b l e b o n d s , n i s a n a b b r e v i a t i o n f o r omega a n d p = t h e p o s i t i o n o f t h e f i r s t d o u b l e bond f r o m t h e m e t h y l e n d of t h e m o l e c u l e . T h e r e f o r e 16:ln7 i s a monoenoic u n s a t u r a t e d f a t t y a c i d w i t h 16 c a r b o n a t o m s , 1 d o u b l e bond a n d t h e p o s i t i o n o f t h e d o u b l e bond i s on c a r b o n 7 c o u n t i n g f r o m t h e m e t h y l e n d .  8 to t h e presence o f nonpolar hydrocarbon groups. groups  i n a s s o c i a t i o n with f a t t y a c i d s , forming  s p h i n g o l i p i d s and o t h e r s ,  However p o l a r phospholipids,  impart v a r i o u s degrees o f water  s o l u b i l i t y t o the molecule.  L i p i d s c a n t h e r e f o r e be c l a s s i f i e d  on t h e b a s i s o f t h e i r p o l a r i t y ,  i e t h e n e u t r a l and t h e p o l a r  lipids.  Neutral  lipids  the major  include the triacylglycerides  lipid  energy depot.  (TAGs), which  TAGs c o n s i s t o f t h r e e  fatty acids esterified to the alcohol, glycerol. particularly 18:ln9, (T)  14:0 a n d 16:0,  c a r b o n o f g l y c e r o l i n TAG m o l e c u l e s .  o f g l y c e r o l may be a t t a c h e d monounsaturate (Christie,  16:ln7 and  (a) and t h i r d  The s e c o n d  (0) c a r b o n  t o another saturate or t o a  o r , a s i s common i n f i s h ,  1986).  long-chain  Saturates,  and t h e monounsaturates,  t y p i c a l l y occupy p o s i t i o n s on t h e f i r s t  form  t o a PUFA o r HUFA  I n t h e second p o s i t i o n , these long  chain  f a t t y a c i d s a r e l e s s s u s c e p t i b l e t o o x i d a t i o n f o r energy (Murray, 1988).  O t h e r common n e u t r a l l i p i d s a r e t h e mono a n d  diacylglycerides  ( w i t h o n e o r two f a t t y a c i d s e s t e r i f i e d t o  g l y c e r o l ) ; c h o l e s t e r o l and c h o l e s t e r y l e s t e r s ( c h o l e s t e r o l esterified  The  polar  t o a f a t t y a c i d ) , and f r e e f a t t y  lipids  acids.  include the phosphoglycerides, g l y c o l i p i d s ,  s p h i n g o m y e l i n s and p l a s m o l a g e n s . g l y c e r o l or sphingosine f a t t y acid residues  T h e s e compounds h a v e e i t h e r a  ( a n amino a l c o h o l ) b a c k b o n e , one o r two  and a p o l a r head group.  of t h e g l y c o l i p i d s a l l o f these p o l a r  With t h e exception  l i p i d s contain a  9 p h o s p h a t e g r o u p and  a r e o f t e n r e f e r r e d t o as  phospholipids.  G l y c o l i p i d s contain sphingosine,  a f a t t y a c i d and  sugar groups.  and  Phosphoglycerides  g l y c o l i p i d s are  c o n s t i t u e n t s of biomembranes.  Phosphatidylcholine  phosphatidylethanolamine  phosphatidylserine  phosphatidylinositol  <PE),  g a n g l i o s i d e s are g l y c o l i p i d s ,  important  i n n e u r a l t i s s u e and  the b r a i n .  of the p h o s p h o l i p i d s  10%  m u s c l e t i s s u e ( M u r r a y et  but are  in brain  and  al. , 1988).  frequently  i n the a - p o s i t i o n of the g l y c e r o l backbone i n p o l a r PUFA o r HUFA i n t h e  p o s i t i o n , the unsaturate  i s protected  monoene i s o x i d i z e d ( G r e e n e and  D i e t a r y f a t t y a c i d s and  Saturated  P-position.  In  i f the s a t u r a t e  this or  S e l i v o n c h i c k , 1987).  requirements f o r growth  f a t t y a c i d s w i t h e v e n numbers o f c a r b o n atoms c a n  synthesized  de  f a t t y a c i d s can  novo f r o m a c e t a t e be e l o n g a t e d  i n t o compounds as r e q u i r e d .  and  (Castell,  i n the d i e t .  1979).  desaturated  and  be  Some d i e t a r y incorporated  Other f a t t y a c i d s cannot  s y n t h e s i z e d , have s p e c i f i c m e t a b o l i c provided  and  plasmolagens  monounsaturated f a t t y a c i d s are  l i p i d s w i t h an n3  2  (PS),  Sphingomyelins  c o m p r i s e a s much a s  present  (PC),  t h r o u g h o u t t h e body  nerve t i s s u e w h i l e  and  important  Cerebrosides  a l s o f o u n d i n b r a i n and  Saturated  o r more  ( P I ) and d i p h o s p h a t i d y l g l y c e r o l  ( c a r d i o l i p i n ) are a l l phosphoglycerides.  especially  one  f u n c t i o n s and  be  must  These a r e t h e e s s e n t i a l f a t t y 1  be  acids.  10 F i s h cannot synthesize linolenic  a n y member o f t h e l i n o l e i c  (n3) s e r i e s u n l e s s  structure i s present.  a precursor  w i t h t h e same omega  In a d d i t i o n , there  i s competitive  i n h i b i t i o n b e t w e e n t h e n 3 , n6 a n d n9 s e r i e s s u c h elongation  Dietary  The n3 s e r i e s i s t h e most p o t e n t  f o l l o w e d by t h e n6 a n d n9 s e r i e s ( H a l v e r ,  s e r i e s ) f a t t y a c i d s f o r rainbow t r o u t .  Castell  calories.  2.7% o f t h e d i e t a r y  T h i s was b a s e d on a c h i e v i n g t h e b e s t  feed conversions  essential  a n d on a l l e v i a t i n g  growth  during  their  heart  studies.  T a k e u c h i a n d Watanabe (1977b) f o u n d t h a t a s t h e t o t a l lipid  increased  l e v e l of  i n t h e d i e t of rainbow t r o u t , t h e requirement  18:3n3 a l s o i n c r e a s e d .  sufficient  rates  several signs of  f a t t y a c i d d e f i c i e n c y , eg. f i n e r o s i o n ,  myopathy and syncope t h a t o c c u r r e d  for  F o r e x a m p l e 1% 18:3n3 was \  i n diets containing relatively  t o 5%) b u t i n d i e t s c o n t a i n i n g 14% l i p i d ,  low l i p i d  l e v e l s (up  the requirement f o r  18:3n3 was more t h a n 2% o f t h e d i e t f o r maximum g r o w t h .  it  i s suggested that the e s s e n t i a l f a t t y a c i d  should for  et  (1972 a , b , c ) e s t a b l i s h e d t h e r e q u i r e m e n t f o r 18:3n3 a t 1%  o f t h e d i e t by w e i g h t o r a p p r o x i m a t e l y  and  1980).  s t u d i e s h a v e d e m o n s t r a t e d t h e e s s e n t i a l i t y o f t h e n3  (linolenic al.  that  a n d d e s a t u r a t i o n o f one s e r i e s i s i n h i b i t e d by t h e  members o f a n o t h e r s e r i e s . inhibitor,  (n6) o r t h e  Hence,  requirement  be e x p r e s s e d a s a p e r c e n t a g e o f t h e d i e t a r y l i p i d a n d  r a i n b o w t r o u t t h e r e q u i s i t e l e v e l was s e t a t  approximately  20% a s 18,:3n3.  Some w o r k e r s for of  b e l i e v e t h a t n6 f a t t y a c i d s may a l s o be e s s e n t i a l  salmonids.  N i c o l a i d e s and Woodall  j u v e n i l e Chinook  (1962) r e p o r t e d 70-80%  salmon e x h i b i t e d dermal  depigmentation  t h e y w e r e f e d d i e t s c o n t a i n i n g e i t h e r no l i p i d , triolein  18:0 a s  a t 1% o f t h e d i e t , o r 0.1% o f t h e d i e t a s 1 8 : 3 n 3 .  When t h e f i s h  w e r e f e d 1% o f t h e d i e t a s 18:2n6 ( a s  trilinolein),  86% d i s p l a y e d normal  pigmentation.  When 0.1%  18:3n3 a n d 1% t r i l i n o l e i n w e r e i n c l u d e d i n t h e d i e t , fish  when  were n o r m a l .  Growth r a t e s were a l s o s t u d i e d and f o u n d t o  be h i g h e s t i n t h o s e f i s h  receiving  d i e t and t h e s e were n o t improved trilinolein)  69% o f t h e  18:3n3 a l o n e a t 0.1% o f t h e  e v e n when 3% 18:2n6 ( a s  was a d d e d t o t h e d i e t .  Ackman a n d T a k e u c h i  ( 1 9 8 6 ) r e p o r t e d l e v e l s o f 18:3n3 a n d 20:4n6  t h a t were t h r e e and t e n t i m e s h i g h e r , r e s p e c t i v e l y ,  i n wild  t h a n i n h a t c h e r y - r e a r e d A t l a n t i c p r e - s m o l t s o v e r w i n t e r i n g a t 00.5° C. which  The h a t c h e r y s m o l t s e x p e r i e n c e d s e v e r e f i n e r o s i o n  often ledto mortality.  T h i s prompted an i n v e s t i g a t i o n  i n t o t h e p o s s i b i l i t y o f an e s s e n t i a l Although the hatchery reared f i s h than t h e w i l d f i s h ,  to  20:4n6.  fatty acid  deficiency.  h a d h i g h e r l e v e l s o f 18:2n6  t h e y d i d n o t e l o n g a t e a n d d e s a t u r a t e 18:2n6  I t was s u g g e s t e d  t h a t h i g h l e v e l s o f n3 f a t t y a c i d s  may h a v e i n h i b i t e d t h e e l o n g a t i o n / d e s a t u r a t i o n o f 18:2n6 i n the hatchery f i s h .  M o r e o v e r , i t was n o t e d t h a t t h e a q u a t i c  12 i n s e c t d i e t of the w i l d f i s h 20:4n6.  T h e r e was  no  c o n c l u s i v e proof  20:4n6 w e r e p r e v e n t i n g Ackman and  included high  f i n erosion  l e v e l s of  that high  l e v e l s of  i n the w i l d f i s h .  T a k e u c h i s u g g e s t e d t h a t 20:4n6, b e i n g  prostaglandins  ( s e e b e l o w ) , may  pre-formed  However  a precursor  have been i m p o r t a n t  to  in a  d e f e n s e mechanism a g a i n s t t h e l e s i o n s .  I t has  a l s o b e e n shown t h a t d i e t a r y i n c l u s i o n o f  prevents  the  accumulation of  20:3n9, an  i n d i c a t o r of e s s e n t i a l  f a t t y a c i d d e f i c i e n c y ( G r e e n e and  Selivonchick,  I t i s g e n e r a l l y b e l i e v e d t h a t the  n6  s e r i e s ) are other  not  1987).  fatty acids  (linoleic  e s s e n t i a l f o r salmonids although  s p e c i e s , p a r t i c u l a r l y the homeothermic  animals.  18:2n6  Castell  they are  land-dwelling  ( 1 9 7 2 a ) d e m o n s t r a t e d t h a t no  combination  18:2n6 and  18:3n3 r e s u l t e d i n a s good a g r o w t h r a t e o r  conversion  as  18:3n3 a l o n e a t 1% o f t h e d i e t .  a l s o showed t h a t t h e b e s t f e d 1%  18:3n3 a s t h e  Yu and  Sinnhuber  kisutch) as n3  depressed growth. acids  acid.  o r h i g h d i e t a r y l e v e l s o f n3 The  (Oncorhynchus  of a d i e t c o n t a i n i n g  p r e s e n c e o f more t h a n 1%  10%  approximately  lipid  n6  fatty  fatty  acids  optimum l e v e l o f p o l y u n s a t u r a t e d  (n3+n6) i n t h e d i e t was  (1979)  when t r o u t w e r e  sole dietary essential fatty  The  2.5%  of  feed  e t al.  (1979b) f o u n d t h a t c o h o s a l m o n  fatty acids.  o r e x t r e m e l y low  Yu  growth r a t e s occurred  f r y r e q u i r e d 1 t o 1.5%  for  or  acids  fatty  less.  13 T a k e u c h i et al.  (1979c) i n v e s t i g a t e d f a t t y a c i d r e q u i r e m e n t s i n  chum s a l m o n  (Oncorhynchus  keta)  f r y h e l d i n b o t h f r e s h and  saltwater.  The b e s t w e i g h t g a i n a n d f e e d c o n v e r s i o n was  o b t a i n e d when b o t h 1% 18:3n3 a n d 1% 18:2n6 w e r e p r e s e n t .  F a t t y a c i d r e q u i r e m e n t s f o r Chinook salmon tshawytscha) cycle. acids of  (Oncorhynchus  have n o t been r e p o r t e d f o r any s t a g e o f t h e i r  life  The p r e s e n c e o f l o n g c h a i n h i g h l y u n s a t u r a t e d n3 f a t t y (n3 HUFAs) a r e a n o t a b l e f e a t u r e o f t h e body c o m p o s i t i o n  a l lPacific  salmon  (Plotnikoff  e t al., 1 9 8 4 ) , a n d j u v e n i l e  C h i n o o k s a l m o n a r e a b l e t o c o n v e r t 18:3n3 t o n3 HUFAs ( M u g r d i t c h i a n e t al., 1982 a n d D o s a n j h e t al., 1 9 8 8 ) . suggests the importance of the l i n o l e n i c  readily This  (n3) s e r i e s f o r t h i s  species.  Signs of e s s e n t i a l  fatty acid deficiency  poor growth; e l e v a t e d fin;  l e v e l s o f 20:3n9; n e c r o s i s o f t h e c a u d a l  fatty, pale l i v e r ;  dermal d e p i g m e n t a t i o n ; i n c r e a s e d water .  c o n t e n t o f t h e muscle; syncope mitochondrial of  liver  i n salmonids include:  swelling;  i n response t o shock;  h e a r t myopathy; i n c r e a s e d  increased  respiration  homogenates; and low hemoglobin l e v e l s ( S i n n h u b e r ,  1969).  3  Digestion of dietary  lipids  Hydrolysis of triacylglycerides, lipids,  t h e m a j o r component o f d i e t a r y  i s c a t a l y s e d by p a n c r e a t i c  lipase.  This results i n the  f  14 l i b e r a t i o n o f f a t t y a c i d s and m o n o a c y l g l y c e r i d e s . acids esterified  t o t h e TAG a r e a l l s a t u r a t e d , t h e n t h e r a t e o f  hydrolysis f o r a l l three positions w i l l p o s i t i o n two i s o c c u p i e d hydrolysed, in  be t h e same.  by a n u n s a t u r a t e  leaving a monoacylglyceride.  p o s i t i o n one o r t h r e e ,  r a t e as a saturated  i t will  If  t h e n i t may n o t be I f the unsaturate i s  be h y d r o l y s e d  f a t t y a c i d (Leger,  Hydrolysis of dietary phospholipids formation  I f the f a t t y  a t t h e same  1985).  i s thought t o r e s u l t i n the  o f f r e e f a t t y a c i d s and l y s o p h o s p h o l i p i d s  mammals ( H e n d e r s o n a n d T o c h e r , 1987).  as i n  Typically polar  lipids  have an e s s e n t i a l f a t t y a c i d e s t e r i f i e d t o t h e 3 p o s i t i o n and a s a t u r a t e o r monoene a t t h e a p o s i t i o n . probably  This  arrangement  makes t h e e s s e n t i a l f a t t y a c i d l e s s s u s c e p t i b l e t o  h y d r o l y s i s a n d r e m o v a l t h a n f a t t y a c i d s i n t h e a p o s i t i o n (Dave et al., 1976, c i t e d  i n C a s t l e d i n e and B u c k l e y ,  Liberated f a t t y acids, monoacylglycerides a r e a b s o r b e d by t h e p r o x i m a l The  rate of absorption  and  1982).  lysophospholipids  i n t e s t i n e and t h e p y l o r i c  i s much s l o w e r  t h a n i n mammals a n d  appears t o i n c r e a s e w i t h t h e degree o f u n s a t u r a t i o n water temperature  caecae.  ( H e n d e r s o n a n d T o c h e r , 1987).  and t h e  A l l dietary  f a t t y a c i d s , except those that a r e r a p i d l y incorporated phospholipids, et al., 1977).  into  a r e e i t h e r c a t a b o l i z e d f o r e n e r g y o r s t o r e d (Yu S i r e et al.  (1981) h a s d e m o n s t r a t e d t h a t  fatty  a c i d s a r e r e - e s t e r i f i e d t o TAGs w i t h i n t h e i n t e s t i n a l e p i t h e l i u m and i t i s t h o u g h t t h a t l y s o p h o s p h o l i p i d s  are r e -  15 e s t e r i f i e d w i t h f a t t y a c i d s t o p h o s p h o l i p i d s as o c c u r s i n mammals ( H e n d e r s o n a n d T o c h e r , 1987).  A f t e r c r o s s i n g t h e i n t e s t i n a l w a l l , most d i e t a r y packaged for (eg.  i n t o c h y l o m i c r o n s a n d some i n t o  t r a n s p o r t by t h e lymph ( L e g e r , 1985).  lipids are  lipoproteins Most  (VLDL)  lipoprotein  VLDL, LDL, HDL) s y n t h e s i s t a k e s p l a c e i n t h e l i v e r i n  salmonids, i n c l u d i n g the l i p o p r o t e i n , v i t e l l o g e n i n , which i s p r o d u c e d by m a t u r i n g f e m a l e f i s h  ( H e n d e r s o n a n d T o c h e r , 1987).  U p t a k e a n d s t o r a g e o f TAGs o c c u r s i n a d i p o s e t i s s u e and S a r g e n t , 1985) a n d i n m u s c l e , p a r t i c u l a r l y  (Henderson  i n thered  m u s c l e i n s a l m o n i d s ( H e p h e r , 1988).  4  Role of l i p i d s  i n fish  Energy i s r e q u i r e d f o r maintenance, growth and r e p r o d u c t i o n i n all  animals.  While f i s h p r e f e r e n t i a l l y c a t a b o l i z e p r o t e i n t o  meet t h e i r e n e r g y r e q u i r e m e n t s , d i e t s f o r f a r m e d f i s h a r e formulated t o minimize p r o t e i n consumption f o r energy. spares c o s t l y p r o t e i n f o r growth and t i s s u e r e p a i r .  This  Salmonids  have a l i m i t e d a b i l i t y t o u t i l i z e c a r b o h y d r a t e as an energy source.  E x c e s s (>20%) d i e t a r y c a r b o h y d r a t e f e d t o r a i n b o w  trout resulted i n enlarged l i v e r s  with glycogen-filled vacuoles  i n t h e h e p a t o c y t e s , d e p r e s s e d growth r a t e s and i n c r e a s e d mortality  ( H i l t o n a n d S l i n g e r , 1981).  therefore incorporates l i p i d s source.  The n u t r i t i o n i s t  i n t o t h e d i e t as t h e major  The m e t a b o l i z a b l e e n e r g y v a l u e o f l i p i d s  i s 9.45  energy  16 kcal/g l i p i d ,  more t h a n d o u b l e t h a t a v a i l a b l e f r o m  carbohydrate  (4.0 k c a l / g f o r d e x t r i n and g l u c o s e ) o r p r o t e i n (4.5 k c a l / g f o r crude p r o t e i n )  (McCallum  and H i g g s ,  1989).  A s i d e from the p r o v i s i o n of energy,  l i p i d s f u n c t i o n i n the  t r a n s p o r t o f t h e f a t s o l u b l e v i t a m i n s , A, D, tissues. provide  They i m p r o v e  t h e p a l a t a b i l i t y o f f o r m u l a t e d d i e t s and  e s s e n t i a l f a t t y a c i d s as d i s c u s s e d  previously.  F a t t y a c i d s a r e i n t e g r a l p a r t s o f many v i t a l body o f a l l a n i m a l s .  The  function in cell  components o f t h e  amphipathic nature (having  and n o n p o l a r c o m p o n e n t s ) o f p o l a r their  E and K, t o a l l  membranes.  both  polar  l i p i d molecules i s c e n t r a l to O r i e n t a t i o n of these  lipids  a t w a t e r - o i l i n t e r f a c e s i s such t h a t the p o l a r group i s i n the w a t e r p h a s e and t h e n o n p o l a r p o r t i o n i s i n t h e o i l p h a s e . b i l a y e r of these p o l a r l i p i d s  A  i s the basic s t r u c t u r e of  b i o l o g i c a l membranes, b o t h c e l l u l a r and s u b c e l l u l a r .  N a t u r a l l y o c c u r r i n g unsaturated f a t t y acids are almost  entirely  o f t h e c i s c o n f i g u r a t i o n and t h e m o l e c u l e i s b e n t  at a  d o u b l e b o n d . As t h e l e v e l o f u n s a t u r a t i o n  120°  increases  in  membranes, t h e m o l e c u l e s p a c k t o g e t h e r l e s s t i g h t l y due greater  numbers o f t h e s e b o n d s a n d t h e c o n c o m i t a n t  v a n d e r Waal a t t r a c t i o n s .  reduction i n  High l e v e l s of u n s a t u r a t i o n  membrane f l u i d i t y and p e r m e a b i l i t y . T h i s important i n cold acclimated  species  to  promote  i s especially  such as t h e  w h e r e i t has b e e n shown t h a t a s t e m p e r a t u r e  salmonids  decreases  a  17 restructuring  o f membranes w i t h  (Hazel,  L i e et a l . , 1989).  1979;  Without a s h i f t associated  t o a more u n s a t u r a t e d  functions  mitochondrial  s u c h as  and  o x i d a t i o n , ATP  as  unsaturation  temperatures  occurs.  production,  gradients, death  The  increased  (Hazel,  would o c c u r  fatty  Leray  e t al.  a c i d composition  28.0%  salt  extent  i n PS  increase important  transport  toward  increase  and  of  of  less in ion  u l t i m a t e l y heat  o f membranes a l s o r e f l e c t s (1984) r e p o r t e d of  PC  f o r 22:6n3 w i t h i n  water.  activity  a d i s s i p a t i o n of  excitability  i n the  r a i n b o w t r o u t , i e f r o m 22.4% to  with  an  of  compromised.  a shift  shift  rates  1979).  dynamic c o m p o s i t i o n  changes.  would be  increase,  neural  catalytic  to cytoplasmic  Without t h i s  membrane p e r m e a b i l i t y  occurs  s t a t e , membrane  nonelectrolyte permeability  Conversely,  unsaturation  carbohydrate transport,  membrane-bound enzymes, n u c l e a r RNA,  greater  to  one  day  a transformation  intestinal  11.9%  f o r 18:0  PI.  and  i n PE  and  the  of  from  10.6%  to a  to  lesser  T h e s e c h a n g e s were r e l a t e d t o a measured  i n membrane f l u i d i t y  and  i t was  i o n t r a n s p o r t mechanisms may  fluidity  to f u n c t i o n i n the  Sheridan  e t al.  suggested  r e q u i r e an  that increase  in  more s a l i n e e n v i r o n m e n t .  ( 1 9 8 5 ) f o u n d an  22:6n3 i n b o t h t h e  mucosa  in  of t r a n s f e r from f r e s h  S i m i l a r c h a n g e s were f o u n d and  salinity  increase  triacylglycerides  and  i n 2 0 : 5 n 3 , 22:5n3 phospholipids  in  and  18 hatchery  reared  same t i m e t h e y  steelhead trout during  smoltification.  noted the r e d u c t i o n i n t o t a l  of the parr-smolt  transformation.  This  At the  body l i p i d s  lipid  typical  d e p l e t i o n was  due  /  to  a d e c r e a s e i n monoenes a n d s a t u r a t e d f a t t y a c i d s r a t h e r  PUFA a n d i l l u s t r a t e s f a t t y a c i d s by  Bell  et al.  s e l e c t i v e r e t e n t i o n o f PUFA o v e r  other  salmonids.  ( 1 9 8 5 ) d e t e r m i n e d t h a t 22:6n3 was e s s e n t i a l f o r t h e  maintenance of healthy g i l l maximus.  than  epithelium i n turbot,  Four d i e t s were f e d t h a t c o n t a i n e d  Scophthalmus  n6 f a t t y  acids  o n l y o r t h a t had 20:5n3 t o 22:6n3 r a t i o s o f 1.8, 2.2 o r 1 3 . 8 . J u v e n i l e t u r b o t w e r e a b l e t o s u r v i v e w i t h 20:5n3 t o 22:6n3 r a t i o s o f 1.8 o r 2.2 b u t c o u l d n o t s u r v i v e on t h e d i e t c o n t a i n i n g o n l y n6 f a t t y a c i d s . (13.8  r a t i o ) also experienced  apparently  unable t o convert  T h o s e on t h e h i g h 20:5n3  high m o r t a l i t y . 20:5n3 t o 2 2 : 6 n 3 .  These f i s h  and t h e  and s e c o n d a r y l a m e l l a e s l o u g h e d o f f  leaving a skeleton of connective in  were  When d i e t a r y  22:6n3 was l o w o r a b s e n t , c h l o r i d e c e l l s d i s a p p e a r e d e p i t h e l i u m of the primary  diet  t i s s u e w i t h masses o f d e b r i s  the i n t e r l a m e l l a r spaces.  The r a p i d t u r n o v e r  of g i l l  e p i t h e l i u m , and p a r t i c u l a r l y t h e  chloride c e l l s with t h e i r convoluted numerous m i t o c h o n d r i a ,  p l a s m a membranes a n d  makes t h e g i l l s  a s e n s i t i v e i n d i c a t o r of  t h e e s s e n t i a l f a t t y a c i d s t a t u s o f f i s h and i l l u s t r a t e s t h e absolute  r e q u i r e m e n t f o r 22:6n3 a s a s t r u c t u r a l component o f  biomembranes.  Langdon and Thorpe (1984) d e m o n s t r a t e d a  19 p r o l i f e r a t i o n and e n l a r g e m e n t o f c h l o r i d e c e l l s on t h e f i l a m e n t s o f A t l a n t i c salmon to saltwater.  (Salmo  salar)  s m o l t s on t r a n s f e r  T h e s e c e l l s p r o d u c e Na^-IC" A T P a s e and  s u c c i n i c dehydrogenase,  gill  gill  k e y enzymes i n o s m o r e g u l a t i o n and i o n  transport.  F a t t y a c i d s h a v e an i m p o r t a n t r o l e a s s u b s t r a t e s synthesis.  Eicosanoids  prostacyclins  comprise the prostaglandins  ( P G I ) , thromboxanes  (TX) and  PGI and TX t o g e t h e r  All  a r e p h y s i o l o g i c a l l y a c t i v e compounds d e r i v e d  pronounced  a r e o f t e n r e f e r r e d t o as  f a t t y a c i d s and c o n t a i n i n g a 5 - c a r b o n  Eicosanoids  a r e e x t r e m e l y p o t e n t and a v e r y effect.  As l i t t l e  c o n t r a c t i o n o f smooth m u s c l e They a r e s y n t h e s i z e d  (PG),  leukotrienes (LT).  PG,  (eicosa-)  for eicosanoid  prostanoids. f r o m 20  carbon  ring.  s m a l l amount h a s  a  a s 1 ng PG/ml c a u s e s t h e i n a n i m a l s ( M u r r a y e t al.,  1988).  w i t h i n membranes, r a p i d l y t a k e n up,  used  and r a p i d l y d e - a c t i v a t e d .  I n mammals, t h e p r e d o m i n a n t 20:4n6, a s shown i n F i g u r e giving  pathway i s from a r a c h i d o n i c 1 (adapted from Murray  r i s e t o the group 2 e i c o s a n o i d s .  acid,  et al.,  G r o u p s 1 and  1988)  3 are  d e r i v e d f r o m 20:3n6 and 2 0 : 5 n 3 , r e s p e c t i v e l y , i n a n a l o g o u s p a t h w a y s ( F i s c h e r and Weber, 1 9 8 4 ) . eicosanoid  The  s u b s c r i p t of the  i n d i c a t e s t h e number o f d o u b l e bonds i n t h e m o l e c u l e  and t h e group t o w h i c h i t b e l o n g s . The  functions of eicosanoids  a r e many and v a r i e d .  group 2 thromboxane s y n t h e s i z e d  TXA  2  (a  i n p l a t e l e t s f r o m 20:4n6) i s a  20  MEMBRANE PHOSPHOLIPID phospholipase 20:4n6 lipoxygenase  cyclooxygenase  LEUKOTRIENES  PROSTANOIDS ( e g . P G I ) & THROMBOXANES ( e g . T X A ) 2  2  f Figure  1: C o n v e r s i o n o f a r a c h i d o n i c a c i d ( 2 0 : 4 n 6 ) t o g r o u p 2 eicosanoids. By a n a l o g o u s p a t h w a y s , e i c o s a t r i e n o i c a c i d (20:3n6) a n d e i c o s a p e n t a e n o i c a c i d ( 2 0 : 5 n 3 ) a r e s u b s t r a t e s f o r t h e s y n t h e s i s o f groups 1 and 3 eicosanoids, respectively, ( a f t e r M u r r a y et al., 1 9 8 8 )  21 potent PGI  2  s t i m u l a t o r o f p l a t e l e t a g g r e g a t i o n and v a s o c o n s t r i c t i o n .  i s p r o d u c e d i n t h e w a l l s o f b l o o d v e s s e l s and i s  a n t a g o n i s t i c t o TXA .  TXA  2  (a group 3 thromboxane, a l s o  3  s y n t h e s i z e d i n p l a t e l e t s b u t f r o m 20:5n3) a l s o s t i m u l a t e s p l a t e l e t aggregation than TXA .  PGI  2  because TXA  3  aggregation.  3  b u t i t h a s much w e a k e r c l o t t i n g  equals  PGI  2  i n a n t i - c l o t t i n g potency  i s weaker, t h e balance  s h i f t s toward  effects but  anti-  I n a d d i t i o n , group 3 p r o s t a n o i d s b l o c k t h e  s y n t h e s i s o f g r o u p 2 p r o s t a n o i d s by i n h i b i t i n g t h e r e l e a s e o f 20:4n6 f r o m membrane p h o s p h o l i p i d s .  This i s thought  b a s i s o f t h e low i n c i d e n c e o f i s c h e m i c h e a r t d i s e a s e , p l a t e l e t aggregation  and p r o l o n g e d  clotting  times  t o be t h e reduced  i n Eskimos  whose d i e t i s h i g h i n f i s h o i l s w i t h h i g h c o n c e n t r a t i o n s o f 20:5n3 a n d l o w i n c h o l e s t e r o l , t r i a c y l g l y c e r i d e s a n d v e r y l o w density  lipoproteins  (Murray  e t al.,  1988).  L e u k o t r i e n e s a r e s y n t h e s i z e d i n l e u k o c y t e s , p l a t e l e t s and macrophages.  They a c t t o p r o m o t e t h e i n f l a m m a t i o n  response i n  i n f e c t i o n s and t h e h y p e r s e n s i t i v i t y r e a c t i o n i n a l l e r g i e s .  In  mammals e i c o s a n o i d s a l s o m o d u l a t e t h e a c t i o n o f h o r m o n e s , r e g u l a t e blood flow t o p a r t i c u l a r organs,  control  ion transport  a c r o s s some membranes, a n d m o d u l a t e s y n a p t i c t r a n s m i s s i o n a l o n g nerve c e l l s  Although  (Murray  1988).  20:5n3 a p p e a r s t o be t h e t h e m a i n p r o s t a g l a n d i n  precursor i n f i s h , of t h e PG  e t al.,  3  there i s l i t t l e  evidence  f o r the production  series or t h e i r effects i n f i s h tissue  (Greene and  22  Selivonchick,  1987).  P G F « i s a p o t e n t in v i t r o s t i m u l a t o r 2  o v u l a t i o n i n brook t r o u t  (Stacey  and G o e t z , 1 9 8 2 ) ; h o w e v e r i t  i s n o t c l e a r what n a t u r a l r o l e t h e PGF at ovulation  (Goetz, 1983).  the production  and t h a t  o f 20:4n6 t o a m u s c l e and  ( p r e s u m a b l y T X A ) ( A n d e r s o n et al.,  and S e l i v o n c h i c k  (1987)  3  may  Greene  be i m p o r t a n t i n  in fish.  22:6n3, o f t e n t h e  i n c l u d i n g s a l m o n i d s , may  20:5n3 f o r PG s y n t h e s i s  (Tinoco,  Greene and S e v i l o n c h i c k ,  1987).  may  1982;  predominant  be r e t r o c o n v e r t e d  to  Yu and S i n n h u b e r ,  1972;  The p r e s e n c e o f 22:6n3 i n  be more t h a n a m e c h a n i s m t o p r o m o t e  a t low t e m p e r a t u r e s and may,  i n f a c t , be a  ( G r e e n e and  local  storage  d e p o t f o r PG p r e c u r s o r s  1987).  O t h e r w o r k e r s have d e m o n s t r a t e d t h e s y n t h e s i s  leukotrienes directly  1981)  3  ( F i s c h e r and Weber, 1 9 8 4 ) .  T h e r e i s some s p e c u l a t i o n t h a t  membrane p h o s p h o l i p i d s  vaso-  t o PGI , the potent vaso-  suggest t h a t PGI  temperature a c c l i m a t i o n  may  1979,  2  d i l a t i n g compound, a s i n man  fluidity  Studies  20:5n3 ( o r i t s p r o d u c t s )  be a p r e c u r s o r  have  demonstrated  3  20:5n3 may  HUFA i n f i s h ,  (1986)  may  o f T X A , f r o m 20:5n3 i n r a i n b o w t r o u t .  i n h i b i t the transformation c o n t r a c t i n g PG  prostaglandins  Kayama et al.  on p l a i c e s k i n i n d i c a t e d t h a t  of  Selivonchick,  f r o m 22:6n3 (German et al.,  of  1986a,  1986b).  The  lipids  i n t h e r e t i n a l membrane o f t h e human e y e c o n t a i n  o r more o f t h e f a t t y a c i d , 2 2 : 6 n 3 . eye change shape  30%  Rhodopsin molecules i n the  i n response t o l i g h t ,  enabling  i m a g e s t o be  23 perceived.  I t i s thought that the f l e x i b i l i t y  of the r e t i n a l  membrane, due t o t h e p r e s e n c e o f 2 2 : 6 n 3 , f a c i l i t a t e s t h e response of rhodopsin  (Castell,  1988).  Tocher and H a r v i e  (1988) f o u n d h i g h  phosphoglycerides  ( P C , PE, PS, and P I ) o f t h e r e t i n a s o f  r a i n b o w t r o u t a n d c o d (Gadus  l e v e l s o f 22:6n3 i n t h e  morhua).  They p o s t u l a t e d  22:6n3 may be r e t r o c o n v e r t e d t o 20:5n3 b e f o r e synthesis.  P I was f o u n d t o h a v e h i g h e r  than the other phosphoglycerides the f i s h  i n t h i s study  i n t h e eye  l e v e l s o f 20:5n3  i n b o t h r e t i n a s and b r a i n s o f  l e a d i n g t o s p e c u l a t i o n t h a t P I may  as a r e s e r v o i r f o r p r o s t a g l a n d i n  5  prostaglandin  No p o t e n t i a l f u n c t i o n f o r p r o s t a g l a n d i n s  was s u g g e s t e d .  that  serve  precursors.  F a t t y a c i d s y n t h e s i s and m o b i l i z a t i o n  The l i v e r  i s the p r i n c i p a l  in fish.  Little  s i t e o f de novo  synthesis takes  place  c o n t r a s t t o t h e s i t u a t i o n i n mammals. a l s o been r e p o r t e d  De novo  i n the adipose  tissue i n  of rainbow t r o u t  1982).  s y n t h e s i s of f a t t y a c i d s from p r e c u r s o r  in the production  synthesis  F a t t y a c i d s y n t h e s i s has  in v i t r o i n t h e o v a r y  (Weigand and I d l e r ,  fatty acid  acetate  results  o f s a t u r a t e d and monoenoic f a t t y a c i d s  e v e n numbers o f c a r b o n a t o m s .  with  F i s h c a n n o t s y n t h e s i z e any  members o f t h e n3 o r n6 f a t t y a c i d s e r i e s b u t , when p r o v i d e d i n the d i e t ,  salmonids can elongate  and d e s a t u r a t e  n3 and n6  fatty  24 a c i d s t o PUFAs a n d HUFAs a s shown i n F i g u r e 2 ( C a s t e l l ,  Rahm a n d Holman  1979).  (1964) f o u n d t h a t by i n c r e a s i n g t h e amounts o f  d i e t a r y 1 8 : 2 n 6 , t h e e l o n g a t i o n a n d d e s a t u r a t i o n o f 18:3n3 t o 2 0 : 5 n 3 , 22:5n3 and 22:6n3 was s u p p r e s s e d i n w e a n l i n g This suppression a high  rats.  c o u l d be d i s p l a c e d i n t h e o t h e r d i r e c t i o n when  l e v e l o f 18:3n3 was f e d , i e t h e e l o n g a t i o n a n d  d e s a t u r a t i o n o f 18:2n6 t o 2 0 : 3 n 6 , 20:4n6 and 22:5n6  was  inhibited.  This  i n h i b i t o r y effect i s apparently  and,  i n salmonids,  Yu a n d S i n n h u b e r  t h a t n3 f a t t y a c i d s a r e more p o t e n t than the reverse.  concentration  ( 1 9 7 6 , 1979a) d e t e r m i n e d i n h i b i t o r s o f n6 m e t a b o l i s m  The m e c h a n i s m o f c o m p e t i t i v e  r e l a t e d t o s u b s t r a t e s p e c i f i c i t y o f t h e enzyme, which desaturates et  al.,  1981).  inhibition i s 66-desaturase,  18:2n6 t o 18:3n6 a n d 18:3n3 t o 18:4n3  I t i s g e n e r a l l y accepted  i e desaturation occurs  i n the order  fatty  acids  1987).  I n h i b i t i o n o f n3 e l o n g a t i o n a n d d e s a t u r a t i o n may a l s o when t h e l e v e l o f s a t u r a t e s i n t h e d i e t i s r a i s e d . and  Watanabe  acid,  n3 > n6 > n9 e x c e p t when  i s a l a r g e p r e p o n d e r a n c e o f n6 o r n9 f a t t y  (Henderson and T o c h e r ,  (Leger  that the preferential  a f f i n i t y o f t h i s enzyme i s f o r t h e more u n s a t u r a t e d  there  dependent  occur  Takeuchi  (1977b) f o u n d t h a t a n i n c r e a s e i n d i e t a r y  laurate  f r o m 4 t o 14% i n c r e a s e d t h e r e q u i r e m e n t f o r 18:3n3 f r o m 1 t o 2% i n rainbow  trout.  25  SATURATED AND  MONOENOIC FATTY ACIDS  ACETATE  14: 0 — > 1 4 : l n 5 - -> 1 6 : l n 5 16 1 0 --> 1 6 : l n 7 - -> 1 8 : l n 7 18: 0 --> 1 8 : l n 9 - -> 2 0 : l n 9  1 1 22: 0  20: 0 --> 2 0 : l n l l  -- > 2 2 : l n l l  --> 2 2 : l n l 3  POLYUNSATURATED FATTY  18:ln9 / \ 20:ln9 18:2n9 \ / 20:2n9  18:2n6 / \ 20:2n6 18:3n6 \ / 20:3n6 / \ 22:3n6 20:4n6 \ / 22:4n6 \ 22:5n6  \  20:3n9  Figure  2:  ACIDS  18:3n3 / \ 20:3n3 18:4n3 \ / 20:4n3 / \ 22:4n3 20:5n3 \ / 22:5n3 \ 22:6n3  Fatty acid synthesis i n f i s h . ( f r o m C a s t e l l , 1979)  V  Competition  26 between s u b s t r a t e s f o r 6 6 - d e s a t u r a s e e x p l a i n s t h e  r e s u l t s o f many o f t h e e a r l y f e e d i n g trout fed diets deficient t h e a d d i t i o n o f 18:2n6.  studies.  Growth r a t e s i n  i n n3 f a t t y a c i d s w e r e i m p r o v e d by However, i f t h e d i e t  contained  a d e q u a t e l e v e l s o f n3 f a t t y a c i d s , t h e a d d i t i o n o f 18:2n6 a t 0.5 o r 1% d e p r e s s e d t h e g r o w t h r a t e .  F u r t h e r m o r e , when 18:3n3  was h e l d a t 0.5 o r 1% a n d 18:2n6 was i n c r e a s e d f r o m 0 t o 5%, the  22:6n3 a n d t o t a l  n3 f a t t y a c i d s i n t h e p h o s p h o l i p i d  f r a c t i o n d e c r e a s e d w h i l e t h e 18:2n6 l e v e l Sinnhuber, 1981).  increased  (Yu a n d '  T h i s i s an e x a m p l e o f c o n c e n t r a t i o n  d e p e n d e n t i n h i b i t i o n o f n3 e l o n g a t i o n a n d d e s a t u r a t i o n by h i g h l e v e l s o f n6 f a t t y  L e g e r et al. as  acids.  ( 1 9 8 1 ) showed t h a t a h i g h  22:6n3 c o u l d e x e r t a n e g a t i v e  l e v e l o f n3 HUFAs s u c h  f e e d b a c k on t h e d e s a t u r a t i o n  o f b o t h 18:3n3 a n d 18:2n6.  M o b i l i z a t i o n and c a t a b o l i s m o f l i p i d s occur when t h e f o o d  supply  i s s c a r c e and d u r i n g t h e spawning  m i g r a t i o n , when w i l d P a c i f i c energy d e f i c i t  1982).  salmon cease f e e d i n g .  When a n  i s p e n d i n g TAGs a r e p r e f e r e n t i a l l y c a t a b o l i z e d  over phospholipids 1980,  during starvation,  (Yu et al., 1 9 7 7 ; C a s t l e d i n e a n d  When p h o s p h o l i p i d s  Buckley,  are catabolized, i ti s l i k e l y  t h a t t h e PUFA o r HUFA i n t h e fl p o s i t i o n a r e p r o t e c t e d  from  o x i d a t i o n and i t i s t h e monoenoic f a t t y a c i d i n t h e a p o s i t i o n t h a t i s b r o k e n down.  Recycling of long chain  between p h o s p h o g l y c e r i d e s  to maintain  unsaturates  vital physiological  27 f u n c t i o n has  b e e n p o s t u l a t e d by C a s t l e d i n e and  C a s t l e d i n e and induce  Buckley  any  crossover  o f PUFAs and  l i p i d pool to phospholipids. in  the n e u t r a l l i p i d  s a t u r a t e s and  monoenes b u t  was  No  phospholipids  HUFAs f r o m t h e  f r a c t i o n w e r e s l i g h t and evident.  p h o s p h o l i p i d c o m p o n e n t , t h e r e was  to  t o determine whether  no  composition  preferential  However i n t h e  a general  decrease i n the  no c a t a b o l i s m o f 18:2n6 o r  t r a n s f e r o f t h e PUFAs and  22:6n3  HUFAs f r o m n e u t r a l  occurred.  M o b i l i z a t i o n also occurs  when n u t r i t i o n a l l y u n b a l a n c e d  d e f i c i e n t d i e t s a r e f e d as  i s the  experimental  f a t t y a c i d content  lipid  neutral  Changes i n f a t t y a c i d  c a t a b o l i s m o f f a t t y a c i d s was  evident.  (1982).  (1980) s t a r v e d j u v e n i l e r a i n b o w t r o u t t o  m o b i l i z a t i o n o f f a t t y a c i d s and  t h e r e was  Buckley  designs.  The  or  i n t e n t i o n i n some of the  neutral  f r a c t i o n o f t h e body l a r g e l y r e f l e c t s t h e c o m p o s i t i o n  dietary  lipid.  Phospholipids  d i e t a r y c h a n g e s and  a r e b u f f e r e d t o some e x t e n t  fatty acids with specific  functions are conserved.  Yu  e t al.  retained disproportionately high  c o n c e n t r a t i o n o f 22:6n3 was  (1977) f o u n d t h a t t r o u t  l e v e l s o f PUFAs and  diets.  In p a r t i c u l a r  high, i n d i c a t i n g the  ( 1 9 8 0 , 1982)  HUFAs i n  levels the  importance  t h i s f a t t y a c i d f o r t r o u t . , S i m i l a r l y when C a s t l e d i n e Buckley  from  metabolic  the p h o s p h o l i p i d f r a c t i o n i n comparison with the employed i n t h e i r e x p e r i m e n t a l  of  and  fed d i e t s deficient i n e s s e n t i a l fatty  a c i d s , n e a r l y c o m p l e t e r e t e n t i o n o f n3 f a t t y a c i d s i n t h e  of  28 phospholipids particularly  was  f o u n d . An  1 8 : l n 9 and  i n c r e a s e i n n9 f a t t y a c i d s ,  20:3n9, was  s e e n as t h e  experiment  progressed.  M e c h a n i s m s may  a l s o e x i s t t o modulate other  (1977) f e d i s o c a l o r i c d i e t s  lipids.  containing various  levels  s a t u r a t e d f a t t y a c i d s t o t h r e e groups of rainbow Regardless  and  constant  i n the t o t a l  (approximately  24%).  of s a t u r a t e s i n c r e a s e d  body l i p i d s  d e s a t u r a t i o n of the  Mobilization process  of  they remained  This suggests r e g u l a t i o n As  the  i n the d i e t , the c o n c e n t r a t i o n  monoenes, p a r t i c u l a r l y 1 8 : l n 9 i n c r e a s e d , i n d i c a t i n g and  al.  trout.  maintainance of a s p e c i f i c degree of s a t u r a t i o n .  level  et  of  of the s a t u r a t e l e v e l s used i n t h e i r d i e t s ,  found t h a t the c o n c e n t r a t i o n fairly  Yu  of  elongation  saturates.  of l i p i d s a l s o occurs  during maturation  and  in  the  vitellogenesis.  y  6  Vitellogenesis  Vitellogenesis  i s the  s y n t h e s i s of y o l k m a t e r i a l s , p a r t i c u l a r l y  v i t e l l o g e n i n , and  t h e i r accumulation  W e i g a n d and  (1982) d e m o n s t r a t e d t h e a b i l i t y o f o o c y t e s  synthesize termed  Idler  lipids early  i n t h e i r development, i n the  'endogenous v i t e l l o g e n e s i s ' .  considered  a m i s n o m e r as  by t h e d e v e l o p i n g  i t i s the  This terminology  oocytes.  period may  be  l i v e r where v i t e l l o g e n i n  s y n t h e s i z e d , as d i s c u s s e d b e l o w , n o t t h e o v a r y .  to  Endogenous  is  29 s y n t h e s i s of y o l k l i p i d s vitellogenesis  p r e c e d e s and  i n time but  terms i s r e l a t i v e l y minor W a l s h , 1988). be  related  The  i t s contribution ( W i e g a n d and  in  exogenous quantitative  I d l e r , 1982;  Mommsen  and  i m p o r t a n c e o f e n d o g e n o u s s y n t h e s i s seems t o  to the p r o v i s i o n  subsequent accumulation 1984).  overlaps  of a s t r u c t u r a l framework f o r  o f y o l k m a t e r i a l s ( F r e m o n t et  I t i s d u r i n g exogenous v i t e l l o g e n e s i s  the  al.,  t h a t the  ovary  g a i n s most o f i t s mass.  The  onset  of v i t e l l o g e n e s i s  i s under hormonal c o n t r o l .  response to the p r o d u c t i o n of a gonadotropin the ovarian f o l l i c l e s  synthesize estrogen  secreted i n t o the systemic primary  t a r g e t f o r 17  synthesis  circulation.  8-estradiol,  and  (exogenous v i t e l l o g e n e s i s )  by t h e p i t u i t a r y ,  hormones w h i c h The  liver  i t responds w i t h  and  export of  Vitellogenin  i s a very high d e n s i t y , female s p e c i f i c  l i p o p h o s p h o p r o t e i n c o m p l e x w h i c h c o n t a i n s c a . 80% essential  1985.).  I t also c a r r i e s carbohydrates,  mineral  salts  (Mommsen and  s e l e c t i v e l y t a k e n up oocytes  including  phosvitin  and  lipovitellin.  Up  t o 87%  of the  and is  developing  i n t o i t s components,  According  (1982), v i t e l l o g e n i n i s t h e m a j o r s o u r c e lipids.  Vitellogenin  i s cleaved  and  (Leger,  phosphate groups  f r o m t h e c i r c u l a t i o n by t h e  by m i c r o p i n o c y t o s i s and  protein  fatty acids  W a l s h , 1988).  the  vitellogenin  W a l s h , 1988).  r i c h i n p h o s p h o l i p i d s and  are  i s the  (Mommsen and  is  In  t o Wiegand  of ovarian p o l a r  l i p i d i n the v i t e l l o g e n i n of  the  30 goldfish,  Carassius  lipovitellin  auratus,  lipid  phospholipid.  i s polar l i p i d  a n d 58% o f t h e  i n c o h o s a l m o n , Oncorhynchus  kisutch,  Early i n vitellogenesis, polar  predominate i n the ovary.  Later, during  lipids  t h e exogenous  TAGs s t a r t t o i n c r e a s e a n d f i n a l l y p r e d o m i n a t e  Circulating  in  stage,  ( W i e g a n d , 1982).  l i p o p r o t e i n s , t h e VLDL, LDL a n d HDL a r e  m a c r o m o l e c u l a r components o f t h e b l o o d vertebrates.  i s  plasma o f a l l  They t r a n s p o r t l i p i d s f r o m t h e s i t e o f a b s o r p t i o n  t h e i n t e s t i n a l mucosa a n d f r o m t h e s i t e o f b i o s y n t h e s i s i n  the  liver  storage core  i n t o t h e c i r c u l a t i o n and t o t h e s i t e s o f c o n v e r s i o n ,  oru t i l i z a t i o n .  Lipoproteins c o n s i s t o f a hydrophobic  o f TAGs a n d c h o l e s t e r o l s u r r o u n d e d b y a h y d r o p h i l i c  envelope o f p o l a r c o n s t i t u e n t s such as p h o s p h o l i p i d s apoproteins  ( M u r r a y et al., 1988) a n d p r o v i d e  materials f o r oogenesis. deposited  directly  transported  Dietary  and  another source o f  l i p i d s a r e t h o u g h t t o be  i n t o t h e eggs d u r i n g  this  by l i p o p r o t e i n s and b y p a s s i n g  period,  the adipose t i s s u e  ( F r e m o n t et al., 1984; L u q u e t a n d W a t a n a b e , 1986).  The  exact  p a r t i t i o n i n g o f v i t e l l o g e n i n and l i p o p r o t e i n , f o r t h e  movement o f y o l k p r e c u r s o r s understood.  i n t o the oocyte i s not w e l l  I t i s thought t h a t both a r e i n v o l v e d perhaps a t  d i f f e r e n t times  ( W i e g a n d , 1982).  L e g e r et al. (1981)  t h a t egg l i p i d s  i n t r o u t a r e f o u n d i n two s e p a r a t e  t h e e g g a n d t h a t t h e y c a n be s e p a r a t e d oil  report  fractions of  by c e n t r i f u g a t i o n . The  g l o b u l e was f o u n d t o c o n t a i n p r i m a r i l y TAGs w i t h t r a c e s o f  31 c h o l e s t e r o l and c h o l e s t e r y l  esters; the yolk globule contained  l i p o v i t e l l i n and p h o s v i t i n .  The n3 f a t t y a c i d c o n c e n t r a t i o n ,  and p a r t i c u l a r l y  2 2 : 6 n 3 , was h i g h e s t i n t h e l i p o v i t e l l i n  was s u g g e s t e d t h a t t h e m a j o r both l i p i d globule.  source of s t r u c t u r a l  and i t  components,  a n d p r o t e i n , f o r t h e d e v e l o p i n g embryo i s t h e y o l k The o i l g l o b u l e i s t h e m a j o r  energy  reservoir  through  incubation.  A v a r i e t y o f o t h e r compounds i s a l s o a c c u m u l a t e d  by t h e g r o w i n g  o o c y t e s , i n c l u d i n g g l y c o g e n , c a r o t e n o i d s , l e c t i n s , wax a n d sterol  e s t e r s , and s i a l o g l y c o p r o t e i n s .  energy  sources, the functions of others i s not c l e a r  and W a l s h ,  W h i l e some o f t h e s e a r e  1988).  Maximum g r o w t h deficiency  and t h e a l l e v i a t i o n o f e s s e n t i a l f a t t y  acid  s i g n s a r e g e n e r a l l y used as t h e c r i t e r i a f o r  e s t a b l i s h i n g d i e t a r y requirements f o r l i p i d essential  (Mommsen  fatty acids.  l e v e l s and  L i t t l e a t t e n t i o n has been p a i d i n  s e t t i n g t h e s e requirements t o p h y s i o l o g i c a l changes such as v i t e l l o g e n e s i s and m a t u r a t i o n .  However, r e c e n t i n f o r m a t i o n  suggests that i n general n u t r i t i o n a l may be q u i t e d i f f e r e n t f r o m f i s h lifecycles. showing  F o r example Luquet  requirements of broodstock  i n the somatic stages of t h e i r a n d Watanabe (1986) r e p o r t w o r k  t h a t m a t u r i n g s a l m o n i d s may be b e t t e r a b l e t o u t i l i z e  c a r b o h y d r a t e t h a n y o u n g e r f i s h due t o t h e p r e s e n c e o f a n amylase  i n t h e i r p y l o r i c caecae.  demonstrated  that diets relatively  T a k e u c h i et al. low i n p r o t e i n  (1981) (33-35%) had  32 no a d v e r s e e f f e c t s o n r e p r o d u c t i o n diet high years g.,  compared w i t h a c o m m e r c i a l  i n p r o t e i n ( 4 3 - 4 7 % ) when f e d t o r a i n b o w t r o u t f o r 3  a s l o n g a s t h e d i e t was a l s o h i g h  i n c l u d i n g 5-7% b e e f t a l l o w ) .  by R o l e y  i n energy  (390 k c a l / 1 0 0  A s i m i l a r r e s u l t was  reported  (1983) who f o u n d t h a t maximum g r o w t h r a t e was  achieved  w i t h a d i e t c o n t a i n i n g b e t w e e n 37 a n d 4 7 % p r o t e i n a n d 3.8 kcal/g metabolizable  e n e r g y b u t t h a t f e c u n d i t y , egg s i z e a n d  embryo s u r v i v a l w e r e u n a f f e c t e d by a p r o t e i n l e v e l 27%  a t t h e same e n e r g y  as low as  level.  F r e m o n t et al. (1984) d e m o n s t r a t e d t h a t r a i n b o w t r o u t w e r e to maintain  high  able  l e v e l s o f 22:6n3 i n v i t e l l o g e n i n a n d  l i p o p r o t e i n when f e d a n n3 d e f i c i e n t d i e t f o r 6 m o n t h s , p r i o r spawning.  Levels of a l l other  to  n3 f a t t y a c i d s , i n c l u d i n g  20:5n3, were r e d u c e d d r a s t i c a l l y d u r i n g t h i s p e r i o d .  The  tenacious  acids  r e t e n t i o n o f 22:6n3 o v e r a l l o t h e r  i n d i c a t e s i t s probable  n3 f a t t y  importance f o r reproductive  a p o s s i b l e d i f f e r e n c e i n requirements f o r growth reproduction  7  i n this  success  and  versus  species.  Embryonic development  From t h e f o r e g o i n g , supply  i t i s c l e a r t h a t salmon embryos r e q u i r e a  o f f a t t y a c i d s a n d p r e f o r m e d compound l i p i d s  <triacylglycerides, phospholipids,  sphingolipids, cholesterol  e t c . ) t o d e v e l o p s u c c e s s f u l l y t h r o u g h t h e i n c u b a t i o n and a l e v i n stages.  These l i p i d s w i l l  form t h e major energy source  as w e l l  33 as  s t r u c t u r a l c o m p o n e n t s f o r g r o w t h and  precursors  compounds o f p h y s i o l o g i c a l i m p o r t a n c e s u c h as prostaglandins. yolk supply of the  has  Deficiency  steroids  of e s s e n t i a l f a t t y a c i d s  d i r e c o n s e q u e n c e s f o r t h e . h e a l t h and  developing  When EFA  for  r a t e s and  sea  abnormalities  L e r a y et  red  sea  b r e a m , low  fecundity  poor h a t c h a b i l i t y r e s u l t e d  case of red  bream, EFA  and  al.  in  the  survival  fish.  d e f i c i e n t d i e t s were f e d t o b r o o d s t o c k o f  t r o u t , c a r p and  and  and  rainbow  fertilization  (Watanabe, 1 9 8 2 ) .  In  deficient diets resulted in  egg  embryo d e f o r m i t i e s .  (1985) o b s e r v e d d e f o r m i t i e s  i n t r o u t e m b r y o s when  b r o o d s t o c k were f e d n 3 - d e f i c i e n t d i e t s w i t h c o n c o m i t a n t l e v e l s o f n6 16-  and  curled  fatty acids.  3 2 - c e l l stage,  disturbed.  L a t e r , the  into helical  Anomalies occurred  when t h e a r r a n g e m e n t o f c e l l s bodies of the  shapes.  L e r a y et  (1985)  precursors  the  was  a l e v i n s were c u r v e d al.  high  as e a r l y as  or  speculated  t h a t t h e s e d e f o r m i t i e s w e r e r e l a t e d t o t h e a b s e n c e o f n3 the  the  of hydroxy f a t t y a c i d s , thought to act  HUFAs,  as  modulators or mediators i n processes of c e l l u l a r  recognition  occurring  Sprecher,  1983;  B o u k h c h a c h e and  1985). also  i n embryonic development  The  L a g a r d e , 1982,  ( A v e l d a n o and cited  i n Leray et  r o l e o f 20:5n3 as a p r o s t a g l a n d i n  implicated.  precursor  al., was  34 Leray  e t al.  times  a n d more r a p i d ( p r e m a t u r e ) y o l k r e s o r p t i o n t h a n i n  control  8  (1985) a l s o r e p o r t e d p r o l o n g e d  embryo d e v e l o p m e n t  f i s h when n 3 - d e f i c i e n t d i e t s were f e d t o  Feeding  regime f o r c u l t u r e d maturing  salmonids  D e p l e t i o n o f a major p o r t i o n o f t h e somatic salmonids latter  occurs  broodstock.  reserves of wild  during v i t e l l o g e n e s i s , p a r t i c u l a r l y  i n the  s t a g e s when f e e d i n g c e a s e s a n d t h e f i s h m i g r a t e  spawning grounds.  This journey  to the  i s l o n g and a r d u o u s f o r s t o c k s  t h a t spawn h u n d r e d s o f m i l e s f r o m t h e o c e a n .  In Chinook  s a l m o n , a l o s s o f c a . 85% o f t h e n e u t r a l l i p i d s , 60-70% o f t h e p h o s p h o l i p i d s a n d 30% o f t h e p r o t e i n f r o m m u s c l e , t h e m a j o r s t o r a g e d e p o t i n t h i s s p e c i e s , was d o c u m e n t e d a f t e r a 4-5 journey  o f more t h a n  River to their  Kato  800 m i l e s f r o m t h e mouth o f t h e C o l u m b i a  spawning grounds (Greene,  (1975) o b s e r v e d  month  1919).  t h a t c u l t u r e d maturing  trout naturally  r e d u c e t h e i r f e e d i n t a k e s e v e r a l months p r i o r t o s p a w n i n g , b u t to  a l e s s e r e x t e n t t h a n t h e w i l d f i s h do.  salmon a l s o reduce t h e i r feed to  spawning but they w i l l  until  i n t a k e over  Cultured  Chinook  s e v e r a l months p r i o r  continue  t o t a k e some f e e d f i g h t  t h e y a r e moved t o f r e s h w a t e r  (personal observation).  Gutsell  up  (1940) f e d t h r e e l o t s o f 3 - y e a r o l d r a i n b o w t r o u t  e i t h e r t o s a t i a t i o n , approximately  50% o f s a t i a t i o n , o r a b o u t  25% o f s a t i a t i o n f o r e i g h t months b e f o r e  spawning.  No  d e f i n i t i o n was the  fish  g i v e n f o r s a t i a t i o n b u t r e f e r e n c e was  f e d t o s a t i a t i o n as b e i n g " o v e r f e d " .  Group 1  ( s a t i a t i o n ) had t h e b e s t f e c u n d i t y on an i n d i v i d u a l f o l l o w e d by g r o u p  2 ( 5 0 % s a t i a t i o n ) and  However 1 0 % o f g r o u p  1 females f a i l e d  decreased i n the o r d e r : group survival group  t o e y e i n g was  t h a t egg q u a l i t y  Ridelman  e t al.  ^  basis  3 (25% s a t i a t i o n ) .  t o spawn.  1 > group  Egg  2 > group  g r e a t e s t i n group  2 ( 7 8 . 7 % ) and t h e n by g r o u p  made t o  size  3.  Percent  3 (80.7%) f o l l o w e d  1 (70.4%).  I t was  by  concluded  s u f f e r e d from o v e r f e e d i n g the broodstock.  (1984) s t a r v e d a group of rainbow  b r o o d s t o c k f o r c a . 40 d a y s p r i o r t o s p a w n i n g .  No  trout differences  i n egg q u a l i t y , q u a n t i t y o r v i a b i l i t y w e r e f o u n d . V i t e l l o g e n e s i s was  a p p a r e n t l y c o m p l e t e , o r n e a r l y s o , by  days p r i o r t o spawning  and  no n e g a t i v e e f f e c t s o f t h e  40  treatment  were e v i d e n t .  S p r i n g a t e e t al. rations to  ( 0 . 7 % o r 0.35%  spawning.  ration.  (1985) f e d rainbow t r o u t e i t h e r f u l l  The  first  o f body w e i g h t p e r d a y ) fish  While a l l of the f u l l - f e d  spawned, c a . 1 1 % o f t h e r e s t r i c t e d g r e a t e r (by 22%)  f i s h of s i m i l a r  fish  The  failed  to  full half  fish  mature.  i n t h e f o r m e r g r o u p b u t when  s i z e s from both groups  f e c u n d i t i e s were s i m i l a r .  prior  t o be s t r i p p e d w e r e t h o s e f e d  s t a r t e d t o spawn.  F e c u n d i t y was  half  f o r a year  T h i s o c c u r r e d a p p r o x i m a t e l y 2-3 weeks b e f o r e t h e  ration fish  larger  or  were compared,  eggs o f t h e f u l l - f e d  i n b o t h d i a m e t e r and w e i g h t and p r o d u c e d  fish  were  larger f r y .  36 Atresia  ( e g g r e s o r p t i o n ) l e v e l s w e r e c a . 7% i n t h e f u l l  group versus  c a . 22% i n t h e h a l f r a t i o n group.  differences i n the t o t a l a c i d and m i n e r a l Fertilization  lipid  i n t h e two g r o u p s .  and s u r v i v a l t o e y e i n g  age w e r e n o t d i f f e r e n t . o b s c u r e d by v a r i a n c e s  T h e r e w e r e no  o r p r o t e i n l e v e l s a n d t h e amino  p r o f i l e s were s i m i l a r  success  ration  Initial  a n d t o 6 months o f  differences i n f r ysize  were  i n g r o w t h i n t h e two g r o u p s a f t e r 4  months.  Scott  (1962) r e p o r t e d t h a t t h e d e g r e e o f f o l l i c u l a r  atresia  increased with the level of starvation i n cultured trout. s i z e was n o t a f f e c t e d by a r e d u c e d f e e d i n g  Egg  regime, but t h e  p r o p o r t i o n o f f i s h w i t h i n a p a r t i c u l a r age c l a s s i n a p o p u l a t i o n t h a t m a t u r e d was r e d u c e d . obtained  by B a g e n a l  studies are conflicting.  t h e r e a p p e a r s t o be no c l e a r c u t b e n e f i t t o f a s t i n g  on t h e r e p r o d u c t i v e p r o c e s s e s  of Pacific  between s t u d i e s i n e x p e r i m e n t a l nutritional  salmon.  Differences  conditions, particularly  b r o o d f i s h c a n be m a i n t a i n e d  without  p r i o r t o s p a w n i n g w i t h no d e t r i m e n t , f e e d and l a b o u r c o s t s would a c c r u e  However  f e e d i n g f o r some p e r i o d  a s i g n i f i c a n t saving i n  t o the farmer.  o r s u r v i v a l c a n be i m p r o v e d by o p t i m i z i n g f e e d i n g before  i n the  h i s t o r y o f t h e f i s h and i n t h e l e v e l o f food  d e p r i v a t i o n , may a c c o u n t f o r t h e d i s s i m i l a r f i n d i n g s . if  were  (1969) w o r k i n g w i t h brown t r o u t .  The r e s u l t s o f many o f t h e p r e c e d i n g At present  Similar results  I f fecundity protocol  spawning, an a d d i t i o n a l advantage i s o b t a i n e d .  37 SECTION  1 - A comparison o f t h e l i p i d acid  profiles  cultured  1.1  composition  and f a t t y  o f t h e eggs o f two s t o c k s o f w i l d a n d  Chinook  salmon.  MATERIALS AND METHODS  1.1.1  E x p e r i m e n t a l d e s i g n and c o n d i t i o n s  Cultured  3 - y e a r o l d C h i n o o k salmon were h e l d  pens a t t h e b r o o d s t o c k r e a r i n g Ltd.  fed  s i t e o f Sea S p r i n g  i n Genoa Bay, n e a r Duncan on V a n c o u v e r  stocks, B i g Qualicum two f o r m u l a t e d  described  later).  i n saltwater net  Island.  (BQ) a n d R o b e r t s o n C r e e k  diets  Salmon Farm Two  ( R C ) , were e a c h  ( d e s i g n a t e d a s COMM o r WV33 a n d  The e x p e r i m e n t a l d e s i g n c o n s i s t e d  of four  pens o f f i s h ,  each c o n t a i n i n g  This  i s a c o m m e r c i a l o p e r a t i o n a n d i t was n o t p o s s i b l e  to  facility  assign  one s t o c k / d i e t c o m b i n a t i o n .  f i s h randomly t o t r e a t m e n t groups because t h e  b r o o d s t o c k were a l r e a d y  i ntheir  this project.  subsampled  sampled  However,  randomly,  as d e s c r i b e d  t h i r d year a t the s t a r t of f i s h f r o m e a c h p e n were  later.  No r e p l i c a t i o n  was  possible.  The  numbers o f f i s h i n t h e p e n s v a r i e d w i d e l y .  fed  t h e COMM d i e t  pens  I n t h e two pens  t h e r e were 149 (RC) a n d 811 (BQ) a n d i n t h e  f e d t h e WV33 d i e t  different  o f pens  t h e r e were 189 (BQ) a n d 283 ( R C ) .  s i z e s o f pens were a l s o u s e d , 680 m  3  Two  a n d 340 m , t h e 3  38 s m a l l e r pens h o l d i n g were.  Not  A l l were below 6 k g /  a l l of the f i s h  September o f t h e i r olds  the f i s h  f e d t h e WV33 d i e t . m. 3  i n t h e pens m a t u r e d third  h a t c h e r y where t h e y were h e l d t o spawning.  held  i n 1988.  were b r o u g h t  with consecutively Manufacturing, dorsal  Fish  During  maturing as 3-year  and t r a n s p o r t e d t o t h e  f o r several  maturation, p r i o r  When t h e f i s h  i n 1987.  summer, t h o s e f i s h  ( 1 9 8 7 b r o o d s t o c k ) were i d e n t i f i e d  f o r spawning  Pen d e n s i t i e s  weeks f o r f i n a l  not maturing  i n 1987 were  t o t h e h a t c h e r y t h e y were  numbered p l a s t i c  Inc.,Seattle,  musculature a n t e r i o r  spaghetti tags  ( F l o y Tag &  Wash., USA), i n s e r t e d  to the dorsal  tagged  i n the  f i n . A l l numbers  were r e c o r d e d and u s e d t o g e n e r a t e t h e random s e l e c t i o n to  be sampled  sampled.  f o r eggs.  Subsequently f i v e  these t e n f o r complete deviation  all  t h r e e were  The  fish  cut  back  were randomly  analysis of their  randomly  selected eggs.  f e m a l e s on t h e COMM d i e t m a t u r e d .  from  A  In t h i s  case  sampled.  were f e d t o s a t i a t i o n  year of t h e i r their  lipid  fish  f r o m e a c h g r o u p were  f r o m t h i s p r o c e d u r e r e s u l t e d when o n l y t h r e e o f t h e  Robertson Creek  of  Ten f i s h  of f i s h  rearing  period  a t Genoa Bay and r e c e i v e d c a . 1%  body w e i g h t p e r d a y . naturally  t w i c e a day d u r i n g t h e t h i r d  During t h e i r  t o c a . 0.5% o f t h e i r  final  summer, t h e y  body w e i g h t  Towards t h e end o f t h e summer t h e y were t a k i n g  even  p e r day. less  feed.  39 Because the pens c o n t a i n e d impossible 1987  t o s t a t e more p r e c i s e l y t h e  b r o o d s t o c k . They w e r e n o t  holding period  One  some f i s h t h a t d i d n o t m a t u r e , i t i s  pen  other  ( c a . 2-4  pen  V a n c o u v e r 33 d i e t (WV33). p r o p r i e t a r y and  provided  on t h e  feed  The not  f o r the  a v a i l a b l e , other  M a l e s and  commercial  than the  information  the  for  i n A p p e n d i x 2 T a b l e A2.1.  f e m a l e s were h e l d  a f e m a l e was  s e p a r a t e l y and  s t a r t of spermiation.  checked r e g u l a r l y was  k i l l e d w i t h a s h a r p blow t o t h e head  and  d i v i d e d among t h e  M i l t was  in  When o v u l a t i o n  egg  o f e g g s r e c e i v e d t h e m i l t f r o m two stock.  the  The  eggs were s t r i p p e d i n t o a c o l l e c t i n g b u c k e t .  c o l l e c t e d and  data  A2.2.  f o r o v u l a t i o n or the evident,  the  West  f o r m u l a t i o n o f t h e West V a n c o u v e r 33 d i e t i s p r e s e n t e d Appendix 2 Table  the  spawning.  formulation  Feed l a b e l  in  freshwater  f e d t h e open f o r m u l a t i o n  label.  commercial d i e t i s given  the  f e d a c o m m e r c i a l d i e t (COMM);  o f e a c h s t o c k was  d i e t was  fed at a l l during  weeks) p r i o r t o  o f e a c h s t o c k was  l e v e l of feeding  taken without  Milt  was  b u c k e t s so t h a t e a c h b u c k e t d i f f e r e n t males of the  killing  the  f i s h and  same  an  i n d i v i d u a l m a l e c o u l d p o t e n t i a l l y be u s e d t o f e r t i l i z e  many  females.  The  eggs o f each f e m a l e were i n c u b a t e d  (Flex-a-Lite Consolidated, and  the  first  egg  i n separate  Tacoma, Wash., USA).  pick, small  Heath t r a y s  After  l o t s o f eggs w e r e o f t e n  eyeing combined.  40 Consequently  d a t a on t h e i n c u b a t i o n  were n o t a v a i l a b l e beyond t h e e y e d  Wild  fish  from each  spawn were sampled Hatchery,  s u c c e s s o f many stage.  stock returning a t B i g Qualicum  by a r r a n g e m e n t  to their Hatchery  and R o b e r t s o n  f o r m a t u r i t y b e f o r e spawning.  eggs o r m i l t .  an egg b u c k e t p r i o r fertilization, No a t t e m p t  Eggs f r o m  to fertilization  however,  by s e v e r a l m a l e s . i n t o Heath  I t was t h e r e f o r e wild  r u n ( a l l age g r o u p s  females impossible to fish.  D a t a on  t o g e t h e r ) were,  old.  In order t o o b t a i n 3-year  olds,  3-year  oldfish  known t o r e t u r n .  T h i s was d e t e r m i n e d  h a t c h e r y managers, Mr. G r a n t Lawseth a t Robertson within  the size  out during  oldfish i s  i n consultation with the  LaDocoeur a t B i g Qualicum  Creek.  4, and  f o r comparison  s a m p l i n g was c a r r i e d  t h a t p o r t i o n o f t h e r u n when t h e peak o f 3 - y e a r  falling  trays.  t o the hatcheries are 3,  salmon r e t u r n i n g  with the cultured  Don  After  available.  Female C h i n o o k 5-years  and s t r i p p e d o f  f e m a l e s were combined i n  success of i n d i v i d u a l  success of the e n t i r e  fish  Ovulating  was made t o keep eggs f r o m d i f f e r e n t  separate a t these hatcheries.  the  several  t h e egg b u c k e t s were e m p t i e d  track the incubation  Creek  i n f r e s h w a t e r r a c e w a y s and  f e m a l e s a n d s p e r m i a t i n g m a l e s were b o t h k i l l e d their  streams t o  When t h e r e t u r n i n g w i l d  r e a c h e d t h e h a t c h e r y , t h e y were h e l d regularly  natal  w i t h t h e D e p a r t m e n t o f F i s h e r i e s and  O c e a n s , Government o f Canada.  checked  individuals  In a d d i t i o n  range o f 3-year  and Mr.  only those  oldfish  were  fish  sampled.  41 S c a l e s were t a k e n c o i n c i d e n t a l l y w i t h egg samples f o r subsequent c o n f i r m a t i o n o f age.  S c a l e s were a g e d by Ms  Y o l e o f t h e A g e i n g U n i t o f t h e Department  Yvonne  o f F i s h e r i e s and  Oceans.  After  the f i s h  were r a n d o m l y complete  1.1.2  lipid  Sample  Approximately within the  were aged, f i v e selected  from a l l  3-year o l d f i s h 3-year o l d f i s h  sampled f o r  collection  50-60 grams o f u n f e r t i l i z e d  ovarian f l u i d ,  placed  eggs were t a k e n  They were d r a i n e d  into plastic  o f most o f  bags and i m m e d i a t e l y  A l l samples were r e - p a c k a g e d , w i t h o u t t h a w i n g ,  oxygen b a r r i e r  bags  into  (W. G r a c e & Co. Canada L t d . , M i s s i s a u g a ,  O n t a r i o ) w i t h i n a week o f c o l l e c t i o n Samples  stock  analysis.  10 m i n u t e s o f s t r i p p i n g .  frozen.  o f each  were a n a l y s e d w i t h i n  f o r s t o r a g e a t -35°C.  10 months o f c o l l e c t i o n .  F o r k l e n g t h and w e i g h t d a t a were t a k e n f r o m a l l f e m a l e s , cultured  and w i l d ,  Approximately  t h a t were  12 h o u r s a f t e r  spawned.  fertilization,  from t h e Heath t r a y o f each c u l t u r e d sampled. (Velsen, to  T h e s e eggs were p u t i n t o 1980) and l a t e r  d e t e r m i n e whether  examined  fertilization  t e n eggs were t a k e n  female t h a t  had been  Stockard's c l e a r i n g  solution  under a d i s s e c t i n g m i c r o s c o p e had t a k e n p l a c e .  Because  42 t h e eggs o f s e v e r a l f e m a l e s a t B i g Qualicum h a t c h e r i e s are p o o l e d a t spawning, n o t be a s s e s s e d f o r t h e w i l d  1.1.3  Egg  and R o b e r t s o n  fertilization  Dry  success could  fish.  composition  A flowchart o u t l i n i n g a l l a n a l y t i c a l procedures Figure  Creek  i s presented i n  3.  weight  determination  S a m p l e s o f c a . 5-10  grams o f f r o z e n e g g s w e r e a c c u r a t e l y  w e i g h e d i n t o e a c h o f t h r e e d r i e d and t a r e d a l u m i n u m p a n s . pans were d r i e d o v e r n i g h t t o c o n s t a n t w e i g h t weight  o f t h e e g g s was  weight. wet  Dry w e i g h t was  weight  of the eggs.  d i f f e r e n c e b e t w e e n 100% of  t h e wet  lipid  by  composition parameters  subtracting the  C.  The  P e r c e n t m o i s t u r e was and  in calculating  A l l formulae used  original  c a l c u l a t e d as  the percent dry weight.  dry  pan  c a l c u l a t e d as a p e r c e n t o f t h e  and d r y w e i g h t s w e r e u s e d  i n the samples.  Lipid  determined  a t 70°  The  The  the  means  percent  for calculating  are p r e s e n t e d i n Appendix  3.  extraction  L i p i d was  e x t r a c t e d from t h e eggs a c c o r d i n g t o t h e p r o c e d u r e  B l i g h and  D y e r (1959) w i t h m o d i f i c a t i o n s by C h r i s t i e  A s a m p l e o f c a . 20 grams o f f r o z e n e g g s was  of  (1982).  a c c u r a t e l y weighed  43  •> d r y m a t t e r & m o i s t u r e determination f r o z e n egg sample •>  lipid extraction ( B l i g h a n d D y e r , 1959)  > TL FAMES  —>  total lipid determination chromatography ( s i l i c a cartridges! CHCl /neutral lipids 3  CH 0H7polar 1ipids 3  > PL FAMES — >  GLC  TLC polar l i p i d determination  Figure  3:  > TLC  Flowchart of procedures f o r l i p i d analysis and f a t t y a c i d d e t e r m i n a t i o n s ( s e e t e x t f o r full description). TL PL FAMES TLC GLC  = = = = =  total lipids polar l i p i d s f a t t y a c i d methyl e s t e r s t h i n l a y e r chromatography gas l i q u i d chromatography  44  and ml and  homogenized  f o r 2 minutes  of methanol.  An a d d i t i o n a l  filtered  apparatus f i t t e d  50 ml o f c h l o r o f o r m were  filtrates added.  was  w i t h a Whatman No.1  cylinder.  latter  solution The  addition  complete,  second  t h e same p r o c e d u r e .  The  KCl s o l u t i o n  poured  i n t o a 500  ml  two  was  graduated  of the KCl caused the s e p a r a t i o n of the and  a c h l o r o f o r m phase,  lipid.  After  removed by a s p i r a t i o n . 20  and a  ml o f 2.6%  the extracted  on a s e c o n d  Buchner  p a p e r . The c o n t e n t s  t h e volume o f t h e c h l o r o f o r m l a y e r  aqueous l a y e r  The  100  i n t o an aqueous p h a s e  containing  repeated  was  and  filter  100  added  seconds.  t o the homogenizer  out using  were combined  The  solution  carried  30  under p r e s s u r e through a  t h e f u n n e l were r e t u r n e d  extraction  the  o f c h l o r o f o r m and  homogenization continued f o r a f u r t h e r  homogenate was  of  w i t h 50 ml  gram sample  the  separation was  measured  This procedure  o f eggs  was  from each  and was  fish  analysed.  The  amount o f l i p i d  layer  of each e x t r a c t i o n  aliquots was  into dried  was  subsamples  of the c h l o r o f o r m  d e t e r m i n e d by p i p e t t i n g  and w e i g h e d  aluminum p a n s .  weight.  overnight  layers  C. and w e i g h e d  The mean amount o f d r i e d  each e x t r a c t i o n  lipid  a t 70°  calculated  3).  The  ml  pans were  to a constant  i n 10 ml was  and u s e d t o c a l c u l a t e  i n e a c h sample, (Appendix  lipid  10  The c h l o r o f o r m  e v a p o r a t e d by g e n t l e warming on a h o t p l a t e .  then d r i e d  for  in triplicate  the t o t a l  determined amount o f  b a s e d on t h e volume o f t h e c h l o r o f o r m Lipid  i n the o r i g i n a l  as a p e r c e n t o f wet  egg  and d r y w e i g h t  sample  f o r each  was  45 extraction  and  the  Separation  of polar  An  a l i q u o t of the  of  lipid  (Buchi  was  lipids  chloroform  the  Sep  Pak  Luer  nitrogen  l o c k of the  gas  lipid  the  poured  lipid  solvent  ml  poured polar  the  open end  lipids  of the  a slight  through the  lipid  stand.  of the  onto the  Rotovapor  oil  tubing  A small  was  Associates, and  attached  stream  tubing  was  of  and  rubber  syringe b a r r e l to  was  top  15%  force  flow  of  fraction  the  first  silica  the  cork  and  60  The  eluent  of the  ml  contained  sample.  of methanol  procedure repeated,  polar  lipids  was  was  the  was  eluting  sample.  o f n e u t r a l and  was  tubing  nitrogen to force  cartridges.  eluted,  of the  hexane i n c h l o r o f o r m  s y r i n g e , and  lipid  s y r i n g e and  from the  separation  plastic  a  mg  cartridges.  loaded  top  neutral  i n t o the  remaining  200-250  b a r r e l of a g l a s s s y r i n g e which  of a s o l u t i o n of  allowing slowly  j o i n e d by  silica  was  on  c a r t r i d g e s (Waters  a retort  non-polar or n e u t r a l  After  The  60  The  a p p l i e d through a p l a s t i c  i n the  i n t o the  attached  on  i n t o the  cartridge,  the  was  assembly  After  silica  Massachusetts)  mounted v e r t i c a l l y  the  phase, c o n t a i n i n g c a .  e v a p o r a t e d under reduced p r e s s u r e  t o two  Milford,  cork  determined.  Rotovapor-R, S w i t z e r l a n d ) .  applied  to  means were  checked  the  46  frequently Plates  by t h i n l a y e r c h r o m a t o g r a p h y  (A. G. Merck, Darmstad,  consisting volume),  (TLC) u s i n g  W. Germany) a n d a s o l v e n t  of hexane-diethy1 ether-formic  according  Silica  acid  G  system  ( 8 0 : 2 0 : 2 by  t o t h e methods o f C h r i s t i e ( 1 9 8 2 ) .  were v i s u a l i z e d w i t h i o d i n e v a p o r a n d s e p a r a t i o n  Bands  was f o u n d t o  be v e r y s a t i s f a c t o r y .  The  polar  three  lipid  eluent  10 ml a l i q u o t s were p i p e t e d  aluminum p a n s ;  of  lipid  weight.  amount o f p o l a r polar  fatty  lipid  entire f i r s t  evaporated neutral  polar  lipid  the weight  (Appendix  3).  The  of polar  esters.  containing  and d r i e d o v e r n i g h t  plus  calculated  i n t h e sample  was u s e d f o r t h e p r e p a r a t i o n  eluent,  of polar  The mean was u s e d t o c a l c u l a t e t h e  lipid  a c i d methyl  overnight  The w e i g h t  i n e a c h a l i q u o t was c a l c u l a t e d by s u b t r a c t i n g  remaining  The  t o a constant  t h e d r y aluminum p a n .  total  i n t o d r y , pre-weighed  e v a p o r a t e d on a warm h o t p l a t e ; d r i e d  ( 7 0 ° C ) ; and w e i g h e d lipid  was made up t o 80 ml w i t h m e t h a n o l and  the neutral  t o a constant  lipids,  weight.  was  The  w e i g h t was u s e d a s a c h e c k on t h e  amount o f l i p i d  originally  applied  to the s i l i c a  cartridges.  Neutral  and p o l a r  lipid  concentrations  were c a l c u l a t e d a s  p e r c e n t a g e s o f t h e d r y weight o f t h e eggs. concentration lipid  Polar  lipid  was a l s o c a l c u l a t e d a s a p e r c e n t a g e o f t h e t o t a l  and as a p e r c e n t a g e o f t h e f a t f r e e d r y weight  of the  47 eggs.  These c a l c u l a t i o n s  Polar this  lipid  determinations  was  d e t e r m i n a t i o n per  acid  Fatty  acid  Since the p r e c i s i o n  satisfactory, sample was  i t was  (of the o r i g i n a l  lipid  was  esterification  used  An  extraction),  to produce t o t a l eluent, after  lipid,  t o make p o l a r l i p i d  used  hexane i n 2 ml c o u l d be  septum v i a l s  analysed  FAME s o l u t i o n s  equipped  silica  by  gas  a Varian  with a flame  a Supelcowax 10  fused  decided  aliquot  that  one  f o l l o w i n g the of  Christie  of the  chloroform  c o n t a i n i n g c a . 50 mg FAMEs.  The  FAMEs.  into  3700 gas  their  liquid  component  Peak a r e a s were i n t e g r a t e d by  (Varian  Instrument  Group, P a l o A l t o ,  mm  a CDS CA,  ID,  The  column  Ontario)  0.25  ym  microprocessor  USA).  fatty  chromatograph  detector.  (30 m x 0.32  film).  until  chromatography.  ionization  column  polar  u n d e r n i t r o g e n below 0 ° C. liquid  of  FAMEs were s t o r e d  ( S u p e l c o Canada L t d . , O a k v i l l e ,  capillary  of  balance  determination of percent  (1 u l ) were s e p a r a t e d  a c i d m e t h y l e s t e r s on  was  these  procedure  lipid  the p o l a r l i p i d  (GLC)  but  sufficient.  sodium m e t h o x i d e .  phase  they  of  m e t h y l e s t e r s (FAMEs) were p r e p a r e d  (1982), using  in  3.  profiles  base c a t a l y z e d methyl  was  i n Appendix  d e t e r m i n a t i o n s were done i n d u p l i c a t e i n i t i a l l y  became e x p e n s i v e .  Fatty  are presented  48 The  GLC  then for  was  temperature  i n c r e a s e d 2° C. 20 m i n u t e s .  220° C.  The  purified  through  injection  r a t i o was  a heated  and  furnace  f o r 10  and  minutes  h e l d a t 230° C.  d e t e c t i o n temperatures  50:1.  remove a l l t r a c e s o f o x y g e n and 22  C.  p e r m i n u t e t o 230° C.  The  split  programmed a t 170°  Helium  carrier  (Supelco  water.  gas  linear  p e a k s were compared w i t h t h o s e  PUFA-1 and  Rapeseed s t a n d a r d s  identified  a c c o r d i n g t o Ackman  than  0.20  included  velocity  % of the t o t a l i n the  (Supelco (1982).  o b t a i n e d by  Canada L t d . )  letter  code.  occurring  Peaks r e p r e s e n t i n g  a r e a o f t h e c h r o m a t o g r a m were  Appendix  eluted.  T h e s e c o d e s were u s e d  The  levels  are 4,  same p o s i t i o n  gained  proceeded.  not  consistently  and  f o r any  peak  and  A2.5  T a b l e s A4.1-A4.8 i n t h e o r d e r t h a t t h e y  were  unidentifiable  l e s s than  carried  0.50  fatty  i n the t o t a l  out  acid profile  2, T a b l e s A2.4  a c i d s were g e n e r a l l y p r e s e n t  %.  and  in duplicate.  by t h e d u p l i c a t i o n  fatty  less  i n t h e chromatograms o f a l l  i n c l u d e d i n Appendix  Fatty acid p r o f i l e s initially  running  profiles.  i n the  s a m p l e s and  the  was  and  U n i d e n t i f i a b l e p e a k s were a s s i g n e d a number o r number  at  to  cm/sec.  Fatty acid  and  was  Canada L t d . )  The  were  was  of e f f o r t ,  polar lipids  were  However, s i n c e l i t t l e  was  a single determination  deemed s u f f i c i e n t  as t h e  analyses  of  49  /  To  avoid f a t t y a c i d o x i d a t i o n during a n a l y s i s , the f o l l o w i n g  precautions  > Egg to  were t a k e n :  samples were k e p t i n t h e f r e e z e r u n t i l  analysis.  p r o c e d u r e and ice  bath  > BHT, to  \  t h e b u c k e t o f t h e h o m o g e n i z e r was  placed  i n an  (50-100 m g / l i t r e ) , was  added  step.  butylated hydroxy-toluene  a l l s o l v e n t s u s e d i n t h e e x t r a c t i o n and  > After evaporation  lipid  silica  was  of s o l v e n t s under reduced p r e s s u r e ,  used to f o r c e l i p i d  cartridges.  t h a t exposure  As  the eluent  l i p i d was  and  of  solvents through  i n c o r p o r a t i o n of a i r i n t o the  the  l e f t the c a r t r i d g e s , the  r u n down t h e s i d e o f  c o l l e c t i o n f l a s k to avoid s p l a s h i n g w i t h the  Diet  the  minimal.  s o l v e n t c a r r y i n g the  1.1.4  Solvents  use.  broken w i t h n i t r o g e n ensuring  t o a i r was  > Nitrogen  separation  i n the p r e p a r a t i o n of methyl e s t e r s .  were f l u s h e d w i t h n i t r o g e n p r i o r t o  vacuum was  prior  F r o z e n eggs were used i n t h e e x t r a c t i o n  during the g r i n d i n g  p r o c e d u r e s and  immediately  the  possible  sample.  composition  A s a m p l e o f e a c h o f t h e two  formulated  d i e t s was  s t o r e d i n o x y g e n b a r r i e r b a g s , as f o r t h e egg  taken  samples.  and  50 To  determine  dry weight  a c c u r a t e l y weighed pans.  slurry  water  i n t h e pans was  was  extracted  applied  t o the eggs,  to  the s t i l l  for  soak  several  Total  determined on  of t h r e e d r i e d  were r e l a t i v e l y  and  then d r i e d  and m o i s t u r e  and  tared  hard,  fatty  following  following  acid  profiles  i t was  out  ml  of The  a t 70°  C  calculated.  necessary  solvent to  i n the t o t a l  i n d u p l i c a t e and  system  grinding.  the procedures f o r the eggs.  t h e d i e t s were c a r r i e d  aluminum  t h e same p r o c e d u r e  i n the f i r s t  them down p r i o r  were  them down.  i n t h e p e l l e t s was  with the e x c e p t i o n t h a t  t o break  5-10  to a constant weight  from the d i e t s  frozen pellets  minutes  lipid  grams o f f r o z e n p e l l e t s  were added t o t h e pans t o b r e a k  mean d r y w e i g h t  Lipid  each  Because the p e l l e t s  distilled  and  into  c a . 5-10  lipid  were  A l l analyses  t h e means were  calculated.  1.1.5  Egg  size  Egg  s i z e was  the  samples  measured i n two  A minimum o f t e n eggs  preserved i n Stockard's s o l u t i o n  taken to monitor calibrated  ways.  fertilization  in millimeters.  (Velsen,  Mean egg  d i a m e t e r was  length divided  Egg  m e a s u r e d by d i s p l a c e m e n t o f w a t e r  by d i v i d i n g  eggs.  d i a m e t e r was  Egg  total  determined  by t h e number o f eggs m e a s u r e d .  A g a i n a minimum o f t e n eggs was determined  1980),  s u c c e s s , were p l a c e d i n a t r o u g h  from the t o t a l volume was  from  measured and  volume d i s p l a c e d  then c a l c u l a t e d  i n a buret.  t h e mean volume by t h e number o f  f r o m t h i s mean.  Egg  diameters determined c o n s i s t e n c y but measurement  1.1.6  by e a c h method were c o m p a r e d f o r  only diameters determined  by  ( f i r s t method) were r e p o r t e d .  Fertilization  Fertilization  and e y e i n g  success  s u c c e s s was m o n i t o r e d by t a k i n g a minimum o f t e n  eggs from a l l f i s h  sampled  12-24 h o u r s a f t e r  T h e s e e g g s were p l a c e d i n S t o c k a r d ' s s o l u t i o n for  direct  clearing.  fertilization. ( V e l s e n , 1980)  Egg c l e a v a g e , i n d i c a t i n g t h a t f e r t i l i z a t i o n  had  o c c u r r e d , was e a s i l y o b s e r v e d by eye o r u n d e r t h e l o w power, o b j e c t i v e of a d i s s e c t i n g  F e c u n d i t y was a p p r o x i m a t e d of  microscope.  a t spawning  e g g s spawned by e a c h f e m a l e .  by e s t i m a t i n g t h e v o l u m e  Eggs a r e t o o f r a g i l e a t t h i s  t i m e t o c o u n t and i t i s n o t a d v i s a b l e t o d r a i n t h e o v a r i a n fluid  s o t h a t a n a c c u r a t e w e i g h t c a n be o b t a i n e d .  At eyeing  t h e e g g s a r e much more r o b u s t and a c o u n t o f t h e l i v e and t h e d e a d e g g s was made f o r e a c h f i s h .  The t o t a l  o f l i v e p l u s dead  e g g s e q u a l s t h e number o f e g g s spawned o r t h e f e c u n d i t y . E y e i n g s u c c e s s was c a l c u l a t e d a s t h e p e r c e n t a g e o f v i a b l e at  t h e eyed  1.1.7  All  s t a g e d i v i d e d by t h e f e c u n d i t y .  Statistical  procedures  d a t a r e p o r t e d a s p e r c e n t a g e s w e r e t r a n s f o r m e d by t h e  arcsine transformation before analyses.  Morphometric  data  eggs  52  (condition  f a c t o r s , egg d i a m e t e r s , e t c . ) ; i n c u b a t i o n  (fertilization parameters polar  rates, eyeing success, e t c . ) ;  (dry matter; moisture; t o t a l  lipid,  results  composition  lipid;  polar  lipid;  f a t free dry weight b a s i s , e t c . ) , i n d i v i d u a l f a t t y  a c i d s , and f a t t y a c i d s e r i e s  (saturates,  all  among t h e t h r e e d i e t s f o r e a c h  analysed f o r differences  s t o c k and f o r d i f f e r e n c e s the  general  of variance (SAS,  1985).  l i n e a r model p r o c e d u r e  f o r each d i e t  using  (PROC GLM) f o r t h e a n a l y s i s  ( 1 x 6 ANOVA), recommended f o r u n b a l a n c e d d a t a Differences  l e a s t s q u a r e means to  between s t o c k s  n 3 , n6, n 9 , e t c . ) were  sets  b e t w e e n means were d e t e r m i n e d by  (LSMEANS) w i t h  t h e PDIFF o p t i o n  i d e n t i f y w h i c h means w e r e d i f f e r e n t .  was s e t a t 95% f o r a l l t e s t s .  (SAS, 1985)  The s i g n i f i c a n c e l e v e l  53 1.2  RESULTS AND  1.2.1  DISCUSSION  Morphometric  measurements  F o r k l e n g t h and w e i g h t d a t a from a l l c u l t u r e d females This  information  i s not r o u t i n e l y taken  hatcheries during  Condition the  factor  formula  but a l l f i s h  this  study  where W - w e i g h t  morphometric data  only as  5 o f t h e 10 f i s h  sampled  diet  2 5  (g) a n d FL = f o r k l e n g t h (cm)  are presented  i n Table  1.  An ANOVA was c a r r i e d  The  o f both  i n the  o u t on a l l t h r e e Because t h e  t«|st r i s k e d  a Type  F o r t h e RC g r o u p s an  t h e RC-COMM f i s h  w o u l d have been v e r y  wild fish  of this  o f t h e BQ-WILD f i s h .  Type 2 e r r o r w o u l d have been  females  i n c l u d i n g BQ-WILD.  ANOVA was p e r f o r m e d e x c l u d i n g test  Unfortunately  sampled a t B i g Q u a l i c u m were 3 y e a r s o l d  g r o u p s o f t h e BQ s t o c k ,  e r r o r i n t h e case  according to  .  sample s i z e was so low, t h e r e s u l t s  this  f o r eggs a t  (1968):  i n d i c a t e d by s c a l e a n a l y s i s and o n l y t h r e e  RC-COMM g r o u p m a t u r e d .  1  a t B i g Qualicum o r  were w e i g h e d and m e a s u r e d .  o f V a n s t o n e and M a r k e r t  3  staff  Salmon Farm.  (CF) was c a l c u l a t e d f o r e a c h f i s h  CF = (W x 1 0 0 0 ) / F L *  All  by t h e h a t c h e r y  spawned a t Sea S p r i n g  Robertson Creek H a t c h e r i e s these  were r e c o r d e d  a s t h e power o f  low ( i e t h e p o s s i b i l i t y  of a  high).  s t o c k s were s i g n i f i c a n t l y  longer (and  54 i  Table  1.  M o r p h o m e t r i c d a t a (mean a n d s t a n d a r d e r r o r o f t h e means) f o r w i l d and c u l t u r e d B i g Q u a l i c u m (BQ) a n d R o b e r t s o n C r e e k (RC) f e m a l e C h i n o o k b r o o d s t o c k . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t BQ WILD mean  Fork  l e n g t h , cm  Weight, kg Condition  factor  BQ WV33 sem (n)  W e i g h t , kg  69.8  5.1"  0.258 (5)  5.1B 0.179 (22)  5.0  3.7-  0.126 (5)  5.1  5.1  factor  fa  0.097 (22)  b  RC WV33 sem (n)  mean  sem (n) b  0.447 (46) 0.124 (46)  b  0.134 (46)  RC" COMM sem (n)  mean  sem (n)  0.547 (12)  67.2 "0.838 (17)  70.7"  1.764 (3)  6.3- •0.124  4.1 «0.166 (17)  4.8"  0.252 (3)  4.8  4.7"  0.152 (3)  80.3  a  (12) Condition  mean  70.1 «0.678 (22)  mean  l e n g t h , cm  sem (n)  77. 8- 0.903 (5)  RC WILD  Fork  mean  BQ COMM  4.1-  0.067 (12)  b  b  b  0.167 (17)  * = The RC-COMM f i s h were e x c l u d e d f r o m t h e ANOVA as t h e sample s i z e was i n a d e q u a t e . See t e x t f o r d i s c u s s i o n . ab •  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between f o r t h e parameter i n d i c a t e d .  stocks  55 heavier  i n t h e c a s e o f t h e RC f i s h )  o r t h e RC-WV33 g r o u p . significantly cultured in  However t h e mean c o n d i t i o n  l o w e r f o r t h e BQ-WILD f i s h  fish.  Condition  t h e RC-WILD f i s h  For s i m p l i c i t y ,  significant  The  were s i g n i f i c a n t l y  b u t t h e r e was no d i f f e r e n c e  By c o n t r a s t ,  t h e RC-WV33 f i s h  differences diet.  1.2.2  The  i n a parameter indicated  Spawning and i n c u b a t i o n  volume o f eggs  stripped  in  h e a v i e r t h a n t h e BQ-WILD  i n length or condition  factor.  i n b o t h l e n g t h and  differences  are too f r a g i l e  group.  success  n o t be d e t e r m i n e d u n t i l t o count u n t i l  Eggs f r o m one o r more f e m a l e s were combined t h e r e were t o o few eggs t o f i l l t h e number o f s u r v i v i n g fecundity  i n condition  f r o m e a c h f e m a l e was measured a t  but fecundity could  individual  by '•'  i n l e n g t h o r w e i g h t between s t o c k s on t h e COMM  s t a g e a s eggs  if  between  follow.  between t h e s t o c k s f o r e a c h d i e t  spawning  lower  T h e r e were no s i g n i f i c a n t  T h e r e were no s i g n i f i c a n t  factors  significantly  were s m a l l e r  w e i g h t t h a n t h e BQ-WV33 f i s h .  f a c t o r was  fish.  g r o u p have been  1 and i n any o f t h e t a b l e s t h a t  RC-WILD f i s h  fish  differences  groups  t h a n f o r t h e BQ  f a c t o r was a l s o  t h a n i n t h e RC-WV33  the s t o c k s f o r each d i e t Table  t h a n t h e BQ c u l t u r e d  then  (Table 2 ) .  a t spawning i f  an i n c u b a t i o n t r a y  eggs was low.  t h e eyed  or a t eyeing  As a r e s u l t ,  d a t a were n o t a v a i l a b l e  f o ra l l the f i s h  56 Table  2.  Spawning d a t a (mean a n d s t a n d a r d e r r o r o f t h e means) f o r w i l d a n d c u l t u r e d B i g Q u a l i c u m (BQ) R o b e r t s o n C r e e k (RC) C h i n o o k b r o o d s t o c k . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD - n a t u r a l d i e t BQ  BQ  WILD mean  BQ COMM  WV33  sem (n)  mean  sem (n)  Volume o f e g g s spawned, ml  no d a t a "  5 9 7 . 6 " 44.8 (21)  Fecundity (# e g g s spawned)  no d a t a "  3 0 1 9 . 9 " 249.2 (12)  Egg d i a m e t e r , mm  7.1-«0.271 (5)  and  7.7* 0.044 (9)  mean  sem (n)  605.4  23.9 (46)  3204.0  92.9 (40)  7.9  b  0.038 (17)  ab = a s i g n i f i c a n t d i f f e r e n c e (a =0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. :  RC  WILD mean  sem (n)  RC  RC COMM  WV33 mean  sem (n)  Volume o f e g g s spawned, m l * *  no d a t a "  3 1 1 . 5" 39.7 (13)  Fecundity"* (# e g g s spawned)  no d a t a "  1 5 9 5 . 6 " 413.6 (5)  Egg d i a m e t e r , mm*"  7.7" 0.061 (12)  * = These d a t a a r e n o t r e c o r d e d **  "  7.7  0.125 (15)  mean  sem (n)  666.7  44.1 (3)  2527.5  294.5 (2)  8.3  0.133 (3)  a t t h e government h a t c h e r i e s  = ANOVA was n o t c o n d u c t e d f o r t h e RC f i s h a s t h e r e was no d a t a f o r t h e WILD f i s h a n d sample s i z e was i n a d e q u a t e f o r t h e COMM f i s h . = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d f o r t h e parameter i n d i c a t e d .  between  stocks  57  t h a t were spawned. observations values  (n) was l o w e r f o r f e c u n d i t y compared w i t h  reported  f o r t h e volume o f eggs spawned  Combined egg l o t s , fecundities,  T h i s e x p l a i n s why t h e number o f  representing the f i s h with  were e x c l u d e d  are  over-estimates,  fish  No  collected  Hatcheries. could  No  o f t h e means.  o f t h e BQ and RC  eggs f r o m more t h a n one  f o rindividual  f i s h are  a t B i g Qualicum o r Robertson  Creek  And b e c a u s e t h e r e were i n s u f f i c i e n t  data,  n o t be c o n d u c t e d on t h e RC  significant  2  (see a l s o T a b l e 3 ) .  f e c u n d i t y or incubation data  routinely  i n t h e case  where c o m b i n i n g  f e m a l e was most p r e v a l e n t  t h e lowest  f o r mean f e c u n d i t y i n T a b l e  particularly  f e d t h e WV33 d i e t ,  (Table 2 ) .  from t h e c a l c u l a t i o n  Consequently the f i g u r e s given  the n  ANOVA  fish.  d i f f e r e n c e was f o u n d  i n f e c u n d i t y , whether  d e t e r m i n e d by volume o r by c o u n t , between t h e BQ-COMM a n d t h e BQ-WV33 f i s h RC  fish  (Table  2).  An ANOVA c o u l d n o t be p e r f o r m e d on t h e  b e c a u s e t h e sample s i z e  f o r t h e RC-COMM g r o u p was t o o  s m a l l a n d t h e r e were no f e c u n d i t y d a t a However an ANOVA between s t o c k s the  BQ-WV33 f i s h  for the wild  f o r t h e WV33 d i e t  were s i g n i f i c a n t l y  more f e c u n d  fish.  revealed  than  t h e RC-  WV33 g r o u p .  These f i g u r e s i n Table combined egg l o t s .  2 do n o t i n c l u d e t h e eggs f r o m t h e  As shown i n T a b l e  that  3, t h e i n c i d e n c e o f  58 Table  3.  Egg r e t e n t i o n a n d o t h e r r e p r o d u c t i v e a b n o r m a l i t i e s o b s e r v e d i n B i g Q u a l i c u m (BQ) a n d R o b e r t s o n C r e e k (RC) c u l t u r e d f e m a l e s on two d i e t s , e x p r e s s e d a s a p e r c e n t a g e o f a l l spawners i n e a c h s t o c k a n d d i e t group. A c t u a l numbers a r e r e p o r t e d i n p a r e n t h e s e s . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33  Stock/Diet  BQ-WV33  RC-WV33  22  Number o f spawners  %  BQ-COMM  17 (#)  %  RC-COMM  46 (#)  3  %  (#)  %  (#)  Level o f retentionslow 31.8 4.5 moderate 4.5 high  (7) (1) (1)  17 .6 (3) 29 .4 (5) 29 .4 (5)  6.5 0.0 0.0  (3) (0) (0)  0. 0 (0) 0. 0 (0) 0. 0 (0)  Spawners n o t r i p e  9.1  (2)  23 .5 (4)  0.0  (0)  0. 0 (0)  Spawners w i t h o n l y one r i p e o v a r y  4.5  (1)  11 .8 (2)  0.0  (0)  0. 0 (0)  Spawners w i t h eggs  0.0  (0)  47 .1 (8)  0.0  (0)  0. 0 (0)  Spawners w i t h no survivors at eyeing  18.2 (4)  41 .2 (7)  2.2  (1)  33. 3 (1)  Spawners w i t h survivors  68.2 (15)  35 .3 (6)  glassy  eyed  97.3 (45)  66. 7 (2)  1 L e v e l o f egg r e t e n t i o n ( u n r i p e e g g s ; n o t o v u l a t e d ) : low = < a c o u n t o f 400 eggs; m o d e r a t e = > a c o u n t o f 400 eggs o r < a w e i g h t o f 400 grams; h i g h = > a w e i g h t o f 400 grams.  59 reproductive  abnormalities  was h i g h ,  WV33 g r o u p s o f b o t h s t o c k s . unripe only clear  (thel e f t )  ovulated,  so a n d n o t r i p e . apparently group.  Unfortunately  egg r e t e n t i o n b u t i t was  and f l a c c i d ,  BQ-WILD f i s h .  are also given  or nearly  Glassy,  i n t h e RC-WV33  t o double or t r i p l e  often pale  i n Table  were s i g n i f i c a n t l y  found  between egg s i z e  i n t h e RC f i s h  i n colour  l a r g e r than those of the had s i g n i f i c a n t l y  T h i s may r e f l e c t  (Blaxter,  n o t be t e s t e d f o r t h e  were a v a i l a b l e b u t c o r r e l a t i o n  between egg d i a m e t e r a n d f e c u n d i t y were c a l c u l a t e d  BQ-COMM and BQ-WV33.  They were -0.1335  r e s p e c t i v e l y b u t were n o t s i g n i f i c a n t Correlation coefficients  (n=15) a n d -0.2550 a t <x = 0.05 ( Z a r ,  were n o t d e t e r m i n e d f o r t h e  RC-COMM and RC-WV33 g r o u p s b e c a u s e f e c u n d i t y d a t a available  t h e lower  a s an i n v e r s e r e l a t i o n s h i p  This could  a s no f e c u n d i t y d a t a  coefficients  The eggs o f t h e  and f e c u n d i t y h a s been r e p o r t e d  Roy a n d H i g g s , 1 9 8 7 ) . fish  2.  I n t e r e s t i n g l y t h e RC-WILD f i s h  eggs t h a n t h e BQ-WILD f i s h .  fecundity  1984).  intact  4, p h o t o g r a p h s ) .  BQ c u l t u r e d f i s h  (n = 6)  fish  o n l y one  d i d n o t mature a t a l l .  w a t e r - h a r d e n e d , eggs were f r e q u e n t  diameter data  1969;  t h e second remained  Some f i s h  s i z e and were s o f t  (Figure  larger  In several f i s h  Some r e t a i n e d eggs were e n l a r g e d  normal  for  d i e t had  t h a t f e c u n d i t i e s were r e d u c e d a n d t h a t t h e RC-WV33  ovary  wild  a t spawning.  were made o f t h i s  were t h e most s e v e r e l y a f f e c t e d .  Egg  among t h e  Many f e m a l e s f e d t h i s  eggs r e t a i n e d on t h e s k e i n s  rough e s t i m a t e s  particularly  f o ronly  2 and 4 f i s h , r e s p e c t i v e l y .  were  60  Figure  4:  Egg a b n o r m a l i t i e s i n R o b e r t s o n C r e e k f e m a l e s on t h e Top: Female w i t h 660 grams o f r e t a i n e d eggs. Bottom: Abnormal eggs a r e p a l e i n c o l o u r , f l a c c i d and e n l a r g e d 2-3 t i m e s n o r m a l .  WV33 d i e t .  61 No  significant  correlations  length, weight o r c o n d i t i o n  (a=0.05) between  f e c u n d i t y and f o r k  f a c t o r were f o u n d i n t h e BQ-COMM  and BQ-WV33 g r o u p s a s h a s been r e p o r t e d by B l a x t e r T h i s may have been b e c a u s e t h e s e f i s h of  egg r e t e n t i o n a s d i s c u s s e d  No  significant  BQ-COMM  (Table 4 ) .  4 0 % o f t h e eggs f r o m t e n f i s h  with  low f e c u n d i t y  (Table  p o o r egg q u a l i t y  the  RC-WV33 f i s h .  3 fish  was a l s o  Fertility  b u t t h e sample  WV33 eggs and a p p e a r e d  s i z e was t o o low t o a s s e s s  greater  i n t h e BQ-WV33 t h a n t h e RCi n t h e BQ-  eggs.  differences  i n t h e number o f e y e d  s u c c e s s ( T a b l e 4) between  Poor s u r v i v a l  b o t h o f t h e WV33 g r o u p s  t h e BQ-COMM  a n d BQ-  r a t e s t o t h e eyed s t a g e o c c u r r e d i n  (Table  3 ) , e g . 7 o u t o f 17 RC-WV33  and 4 o u t o f 18 BQ-WV33 spawners  eyeing.  ( 6 9 % o f t h e eggs o f  ( n o t t e s t e d ) t o be g r e a t e r  T h e r e were no s i g n i f i c a n t  at  fish  t h e RC-COMM a n d t h e RC-WV33 f i s h .  COMM t h a n i n t h e RC-COMM  spawners  together  g r o u p s and p a r t i c u l a r l y i n  low i n t h e RC-COMM  was s i g n i f i c a n t l y  WV33 e g g s .  This  3) s u g g e s t t h a t p o o r p e r f o r m a n c e was due  between  eggs o r i n e y e i n g  I n t h e RC-WV33 f i s h ,  were f e r t i l e .  i n t h e WV33 d i e t  were f e r t i l e )  any d i f f e r e n c e  between t h e  ( T a b l e 2) and o b s e r v a t i o n s o f spawning  to  Fertility  degrees  above.  only  abnormalities  exhibited varying  d i f f e r e n c e was f o u n d i n f e r t i l i t y  a n d BQ-WV33 g r o u p s  (1969).  h a d no s u r v i v i n g  As e x p e c t e d , t h e RC-WV33 g r o u p p r o d u c e d  eggs  62 Table  4.  I n c u b a t i o n d a t a (mean a n d s t a n d a r d e r r o r o f t h e means) f o r w i l d and c u l t u r e d B i g Q u a l i c u m (BQ) and R o b e r t s o n C r e e k (RC) C h i n o o k b r o o d s t o c k . D i e t c o d e s : COMM = c-o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t BQ WILD mean  Fertility,  Eyed  %  no d a t a "  eggs, #  no d a t a "  mean  sem (n)  8 3 . 6 " 7.20 (8) s  mean  sem (n)  97.8-  2.20 (7)  2406.3 - 168.5 (44)  1940.2-" 353.4 (16)  73.9-" " 4.46 (44)  62.4-"" 9.88 (16)  90.87"  Eyed, % ab  sem (n)  BQ COMM  BQ WV33  = a s i g n i f i c a n t d i f f e r e n c e ( a =0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. RC WILD mean  Fertility, Eyed  sem (n)  RC WV33 mean  sem (n)  mean  10.7 (10)  68.9  sem (n) 26.3 (3)  %"""  no d a t a "  e g g s , #"""  no d a t a "  706.9" 272.1 (8)  2392.5  280.5 (2)  90.26"  36.3"" 14.39 (8)  94.7""  0.070 (2)  Eyed, %  40.1"  RC COMM  * = F e r t i l i t y d a t a n o t a v a i l a b l e a s t h e eggs o f s e v e r a l f i s h a r e combined a t spawning. Eyed f i g u r e s f o r t h e s e f i s h a r e f r o m t h e r e s p e c t i v e h a t c h e r y r e c o r d s a n d were n o t i n c l u d e d i n t h e ANOVA. **  ***  •  = D a t a f o r i n d i v i d u a l f i s h were n o t a v a i l a b l e a f t e r as s m a l l egg l o t s were c o m b i n e d .  eyeing  = ANOVA was n o t c o n d u c t e d f o r t h e RC f i s h a s t h e r e was no d a t a f o r t h e WILD f i s h a n d sample s i z e was i n a d e q u a t e f o r t h e COMM f i s h . = a significant difference stocks f o r t h e parameter  (a=0.05) was f o u n d indicated.  between  63 s i g n i f i c a n t l y fewer eyed eggs than the BQ-WV33 f i s h t h e r e was no s i g n i f i c a n t d i f f e r e n c e i n the eyed between these two groups.  although  percentage  The f i g u r e s f o r the RC-COMM f i s h i n  Table 4 a r e f o r 3 f i s h o n l y (only 3 females matured i n t h i s group) and should not be regarded as an a c c u r a t e assessment o f the success o f these  fish.  At the time o f eyeing, dead eggs were removed from the t r a y s and more groups of s u r v i v i n g eggs were combined t o keep the egg trays f u l l .  Data f o r i n d i v i d u a l f i s h were t h e r e f o r e l o s t  beyond the eyed stage.  T h i s meant t h a t s u r v i v a l t o ponding  c o u l d not be estimated a c c u r a t e l y , even on a group b a s i s . Beyond eyeing, however, m o r t a l i t y among a l l groups was low (D. Groves, p e r s . comm.). r  The v a l u e s f o r s u r v i v a l t o eyeing f o r the BQ-WILD and RC-WILD f i s h i n Table 4 a r e f o r a l l age groups spawned throughout the e n t i r e run.  Three year o l d f i s h a r e g e n e r a l l y somewhat l e s s  s u c c e s s f u l than f o u r year o l d s and the m a j o r i t y of spawners a t Big  Qualicum  and Robertson  (G. Ladocoeur  1.2.3  and D. Lawseth, p e r s . comm. and B l a x t e r , 1969).  Composition  Composition  Creek H a t c h e r i e s a r e f o u r year o l d s  of the eggs  data f o r the eggs o f BQ and RC c u l t u r e d and w i l d  broodstock a r e presented i n Tables 5a & 5b. The eggs of a l l groups of both s t o c k s , except RC-WV33, c o n t a i n e d c a . 40% d r y  64 Table  5a.  C o m p o s i t i o n o f t h e eggs (mean a n d s t a n d a r d e r r o r o f t h e means) o f BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n=5 f o r e a c h g r o u p ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t BQ WILD mean  BQ WV3 3 sem  mean  BQ COMM sem  mean  sem  Dry m a t t e r % Moisture %  39 .47 60 .54  0 .75 0 .75  39 .64" 60 .38*  0 .24 0 .25  39. 32 60. 68  0 .49 0 .49  Lipid, Lipid,  % ww" % dw"  12 .19 30 .89  0 .33 0 .62  11 .82 29 .81  0 .17 0 .44  12. 33 31. 37  0 .20 0 .35  P o l a r l i p i d , % dw Neutral l i p i d , % dw  12 .78 18 .83  0 .17 0 .56  12 .84 17 .51  0 .27 0 .56  13. 24 18. 95  0 .30 0 .43  Polar l i p i d , % of total 1 ipid  41 .33  0 .40  43 .14  0 .41  42. 18  0 .59  Neutral lipid, % of total 1 i p i d  61 .41"  0 .66  58 .79  0 .58  60. 38«  0 .85  Polar l i p i d , % o f f a t f r e e dw  19 .35  0 .55  18 .58  0 .45  19. 75  0 .51  Protein  27 .28  0 .59  27 .80"  0 .27  26. 99  0 .37  + a s h ,%  WW  *ww = wet w e i g h t ; ab •  dw = d r y w e i g h t  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 5b.  65 Table  5b:  C o m p o s i t i o n o f t h e eggs (mean and s t a n d a r d e r r o r o f t h e means) o f ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n=5 f o r e a c h g r o u p e x c e p t RC-COMM where n=3). D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t RC WILD  mean  RC WV33  sem  mean  RC COMM  sem  mean  sem  Dry m a t t e r % Moisture %  40 .27- 0. 26 59 .74- 0. 26  35 .89° • 1 .70 64 .11° • 1 .70  39 .59- 0 .97 60 .41- 0 .97  Lipid, Lipid,  11 .98 0. 13 29 .75- 0. 30  11 .54 0 .32 32 .32° 0 .83  12 .47 0 .11 31 .53- 0 .69  P o l a r l i p i d , % dw N e u t r a l l i p i d , % dw  12 .75 17 .11  0. 22 0. 35  14 .16 19 .05  0 .55 0 .57  13 .91 18 .00  0 .34 0 .78  Polar l i p i d , % of total 1 ipid  42 .88  0. 86  43 .76  0 .77  44 .11  0 .70  Neutral l i p i d , % of total l i p i d  57 .50" 0. 93  58 .19  0 .63  57 • 06« 1 .73  Polar l i p i d , % o f f a t f r e e dw  18 .37  0. 42  20 .86  1 .08  20 .06  0 .81  P r o t e i n + a s h .%  28 .01  0. 33  24 .36" 1 .42  27 .12  0 .92  % ww* % dw"  WW  *ww = w e t w e i g h t ; dw = d r y w e i g h t ab  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same r o w .  • = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n stocks f o r t h e parameter i n d i c a t e d . See T a b l e 5 a .  66 matter  and (by s u b t r a c t i o n ) 6 0 % m o i s t u r e .  dry matter  was s i g n i f i c a n t l y  broodstock  than  lower  The c o n c e n t r a t i o n o f  i n t h e eggs o f t h e RC-WV33  i n e i t h e r o f t h e o t h e r RC g r o u p s  o r i n t h e eggs  o f t h e BQ-WV33 b r o o d s t o c k .  Sinnhuber  ( 1 9 6 9 ) h a s r e p o r t e d an i n c r e a s e i n w a t e r c o n t e n t i n  muscle o f salmonids  fedessential  However EFA d e f i c i e n c y h i g h e r water significant  levels  fatty  i n t h e RC-WV33 e g g s .  difference  acid  greater  As  i n n3 f a t t y  in  lipids  this  involved  and s u r v i v a l  group  than  i n t h e s e groups  was no lipids  eggs.  had a h i g h l e v e l o f  Fecundity  rate to eyeing  i n RC-COMM,  i n the  was s i g n i f i c a n t l y  i n t h e RC-COMM  a p p a r e n t l y water hard eggs.  fertility  diets.  ( T a b l e 6b) and t h e n3  concentration of the polar  i n t h e RC-WV33 eggs t h a n  There  acids of the t o t a l  shown i n T a b l e 3, t h e RC-WV33 g r o u p  glassy,  deficient  seems n o t t o have been a f a c t o r  between t h e RC-WV33 and RC-COMM eggs fatty  acid  (Table 2 ) ,  ( T a b l e 4) were a l l l o w e r  a l t h o u g h t h e sample  were t o o low t o a s s e s s  sizes  statistical  significance.  The  m o i s t u r e v a l u e s f o r t h e groups  ( o t h e r t h a n RC-WV33) a r e  i n t e r m e d i a t e between t h o s e g i v e n f o r c u l t u r e d 66.1%) and w i l d and  presented  content and  this  eggs  ( 5 7 . 7 % ) by G r o v e s  i n Appendix  1.  fish  (62.0 -  ( 1 9 8 7 , unpub. r e p o r t )  The d i f f e r e n c e  i n t h e eggs o f t h e w i l d  eggs  i n moisture  between t h e p r e v i o u s  s t u d y may have been due t o s t o c k e f f e c t s .  The  study  67  previous  study  involved  The  dry matter  44%  was p o l a r  wild  fish  f r o m t h e Chemainus  i n a l l g r o u p s was c a . 3 0 % l i p i d  lipid  River.  o f which c a .  and c a . 5 7 - 6 1 % was n e u t r a l  lipid.  41-  On a f a t  f r e e d r y w e i g h t b a s i s , c a . 1 8 - 2 1 % o f t h e d r y m a t t e r was p o l a r lipid.  T h e r e were no s i g n i f i c a n t  groups o f e i t h e r stock were s i g n i f i c a n t between s t o c k s  i n these parameters.  differences  greater  although not s t a t i s t i c a l l y more n e u t r a l report)  lipid  found t h a t  lipid  across  The  higher  lipid  of c u l t u r e d Groves  levels  fish  lower c o n d i t i o n  no  significant  ( or w i l d ,  Groves  ranged from  and w i l d  found  (Tables  fish  ( 1 9 8 7 , unpub. ranged eggs  11.5-12.5%  5a & 5 b ) .  coupled with the better  factors i nthewild  fish  fish,  fish  reported  x  lead t o speculation broodstock'.  egg l i p i d  here,  However i n t h i s  levels  (Table  The  and g r e a t e r egg lend  some  study, there  d i f f e r e n c e s among any o f t h e BQ g r o u p s ,  i ntotal  from  a s r e p o r t e d by  do n o t t e n d t o be p r o d u c t i v e  suggestion.  contained  i n b o t h t h e m u s c l e a n d t h e eggs  compared t o t h e w i l d  diameters i n the cultured support t o t h i s  l e v e l s and,  (wet w e i g h t b a s i s )  performance o f t h e w i l d  'fat fish  lipid  BQ-WV33 eggs a l s o  study they  ( 1 9 8 7 , unpub. r e p o r t )  reproductive that  greater,  a l l groups, c u l t u r e d  there  concentration  eggs and 6.0-13.2% i n w i l d  1) b u t i n t h i s  diet  i e . t h e BQ-WILD and BQ-COMM  neutral  values  However,  lipid  t h a n t h e RC-WV33 e g g s .  13.5-19.0% i n c u l t u r e d (Appendix  i n neutral  on t h e same d i e t ,  eggs had s i g n i f i c a n t l y  d i f f e r e n c e s between  5 a ) . The  were  cultured  68 significantly WV33 eggs moisture lipid BQ  The  higher  lipid  level  (Table 5b) appeared levels  levels  found  (dw) d e t e r m i n e d  f o r t h e RC-  t o be a r e f l e c t i o n  of the high  i n these  eggs.  were s t a t i s t i c a l l y  The RC-WILD and RC-COMM  no d i f f e r e n t  than  those  of the  eggs.  data of Tables  Since the level t h e s e eggs,  5 a a n d 5b a r e p r e s e n t e d  of carbohydrate  % protein  i s likely  + % ash (composite  as p e r c e n t a g e s . t o be v e r y  value)  low i n  c a n be  calculated as:  100 % - [% m o i s t u r e  Coincident RC-WV33 There  with the s i g n i f i c a n t l y  eggs was a s i g n i f i c a n t l y  were no o t h e r d i f f e r e n c e s  concentration the  between d i e t  two s t o c k s f o r e a c h  fatty  eggs f o r e a c h A4.1 and A4.2. of  fatty  acid  i n the protein  of the t o t a l  profiles  + ash  s t o c k n o r between  f o r selected  acids are included here.  fatty  RC-WV33  lipids  of the t o t a l  are presented  i nthe  + % ash value.  BQ-WV33.  s t o c k by d i e t Data  % protein  than  composition  complete  lower  level  with the exception that  1.2.4  acid  higher moisture  diet  lower  Fatty  wet wt]  groups o f e i t h e r  was s i g n i f i c a n t l y  The  + % lipid,  l i p i d s i nthe  i n Appendix  4, T a b l e s  a c i d s and a s s e m b l a g e s  69 Because t h e b i o s y n t h e s i s  acids  were g r o u p e d a c c o r d i n g l y  As shown  higher  significantly  and c u l t u r e d  higher  were s i g n i f i c a n t l y cultured  (by 6-9%)  levels of saturates.  The n6 f a t t y  lower  (by 4-6%)  i n the wild  fish  acids  eggs t h a n i n t h e  significantly  than i n t h e c u l t u r e d  on two d i e t s  eggs.  were acids,  i n t h e BQ-COMM g r o u p a n d more  T h e r e were no s i g n i f i c a n t d i f f e r e n c e s  RC e g g s .  s e r i e s between t h e two  The BQ-COMM eggs had a  higher concentration T h e r e were no o t h e r  these f a t t y a c i d  s e r i e s between  r a t i o o f n3 t o n6 f a t t y a c i d s  frequently  were a l s o  n 3 , n 6 , o r n9 f a t t y a c i d  i n the cultured  RC-COMM e g g s .  i n the wild  d i f f e r e n t i n n 3 , n6 and n9 s e r i e s o f f a t t y  n3 i n t h e BQ-WV33 g r o u p .  statistically  the difference  eggs a l s o h a d  w i t h more n6 and n9 f a t t y a c i d s  saturates,  fish,  g r o u p s b e i n g c a . 7-13% i n t h e BQ  The n9 f a t t y a c i d s  eggs.  significantly  The  analysed  i n the total  The w i l d  BQ eggs f r o m t h e c u l t u r e d  diets  2), the  o f b o t h s t o c k s had  l e v e l s o f n3 f a t t y a c i d s  eggs and 11-13% i n t h e RC e g g s .  in  fish  o f t h e i r eggs t h a n t h e c u l t u r e d  between t h e w i l d  The  and s t a t i s t i c a l l y  i n T a b l e s 6a & 6 b , t h e w i l d  significantly  lower  (Figure  i n fish  6-8).  (Tables  lipids  fatty acids  t h e n 3 , n6 and n9 omega s e r i e s  proceeds within fatty  of polyunsaturated  o f n6 f a t t y a c i d s  than t h e  significant differences i n  stocks.  i n muscle  l i p i d s has  been u s e d a s an i n d i c a t i o n o f t h e e s s e n t i a l f a t t y  70 Table  6a.  S a t u r a t e d , n3, n6 a n d n9 f a t t y a c i d s e r i e s a n d n3:n6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r each g r o u p ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t .  BQ WILD mean S^  BQ WV33 sem  mean 9-6  sem  BQ COMM mean 8-6  sem  Saturates  19.40-  0.38  1 8 . 0 0 ° 0.23  1 8 . 6 6 ° 0.21  n3  39.95-  0.87  3 2 . 3 0 ° 0.59  2 7 . 2 2 ° 0.35  2.34- 0.07  6.61° 0.06  8.47°«0.22  2 8 . 0 1 ° 0.75  3 0 . 3 3 ° 0.36  n6 n9  21.71-  abc •  0.84  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (<x = 0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 6b. BQ WILD mean  n3:n6 r a t i o "  17.07  BQ WV33  BQ COMM  sem  mean  sem  mean  sem  0.58  4.89  0.07  3.21  0.12  * ANOVA was n o t c o n d u c t e d on t h e n3:n6 r a t i o s .  71 Table  6b.  S a t u r a t e d , n 3 , n6 a n d n9 f a t t y a c i d s e r i e s a n d n3:n6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3 ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  RC WILD mean  RC WV33 sem  %  mean  sem  %  RC COMM mean  sem  %  Saturates  1 9 . 9 5 - 0.32  18.69° 0.24  18.21° 0.01  n3  4 1 . 5 6 - 0.59  30.38° 0.47  28.82° 0.18  n6  2.19- 0.13  6.97° 0.07  8.12°«0.14  n9  2 1 . 8 8 - 0.77  29.04° 0.43  29.87° 0.28  ab •  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 6 a .  RC WILD mean n3:n6  ratio'  18.98  RC WV33  RC COMM  sem  mean  sem  mean  sem  0.95  4.36  0.07  3.55  0.07  * ANOVA was n o t c o n d u c t e d on t h e n3:n6  ratios.  72 acid  s t a t u s o f salmonids (eg. C a s t e l l ,  (1964, c i t e d in  i n Castell,  t h e muscle o f w i l d  difference  i n t h e n3:n6 r a t i o s  between  The n3:n6  ratio  i n the total  The  ratio  i n the t o t a l  The n3:n6  ratio  ( T a b l e 6a & 6 b ) .  i e . 17.07 i n BQ eggs  In c o n t r a s t ,  fish  i n the  fish.  i n t h e BQ eggs was c a . 3 t o 5 t i m e s g r e a t e r i n  wild  for  BQ-WILD eggs b u t o n l y  3.55  t h e n3:n6  l i p i d s o f t h e eggs o f a l l c u l t u r e d  the  eggs.  lipids  f i s h was f o u n d t o be c a . 2.6 t o 2.8  ( b o t h s t o c k s ) was c a . 27 - 5 2 % l o w e r t h a n t h e l e v e l muscle o f w i l d  o f 6.69  C h i n o o k m u s c l e and  times g r e a t e r than Gruger's muscle r a t i o , 18.98 i n RC eggs  ratio  C h i n o o k salmon i n s e a w a t e r .  t h e eggs o f t h e w i l d  and  G r u g e r e t al.  1 9 7 9 ) r e p o r t e d an n3:n6  C h i n o o k eggs i s s t r i k i n g . of  1979).  f i s h than i n the c u l t u r e d  Similarly,  fish  (Table 6 a ) , eg.  17.07  3.21 f o r BQ-COMM a n d 4.89 f o r BQ-WV33  i t was 18.98 f o r t h e RC-WILD eggs b u t o n l y  and 4.36 f o r RC-COMM and RC-WV33 eggs r e s p e c t i v e l y  (Table  6b) .  The n3 s e r i e s o f f a t t y  acids  f u r t h e r b r o k e n down i n t o d o u b l e b o n d s ) , HUFAs  i n the total  n3 PUFAs  (n3 f a t t y  bonds) a n d t h e t h r e e f a t t y  l i p i d s h a s been  (n3 f a t t y  acids with  2 to 4  a c i d s w i t h 5 o r more d o u b l e  acids,  2 0 : 5 n 3 , 22:5n3 and 22:6n3 i n  T a b l e s 7a & 7b.  Over a l l groups i n both was  relatively  stocks,  t h e n3 PUFA p r e s e n t  low a t 2-3.5% o f a l l f a t t y  acids  i n t h e eggs  i n the t o t a l  73  Table  7a.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) and t o t a l n3 HUFAs ( h i g h l y u n s a t u r a t e d f a t t y a c i d s ) (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r each group). D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t .  BQ  BQ  WILD mean  BQ  WV33 sem  %  mean  sem  COMM mean  %  sem  %  20:5n3  13.70-"0.36  8.88° 0.16  7.45°  22:5n3  5.49--0.23  3 . 4 7 ° 0.08  2.74°»0.15  22:6n3  17.09- 0.74  17.73- 0.39  2.92- 0.11  1.91° 0.05  1.91° 0.07  36.28- 0.73  30.08°«0.56  25.06°"0.38  Total  n3 P U F A s  Total  n3 H U F A s  1  2  14.87°  1  n3 PUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  2 to 4  2  n3 HUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  5 o r more  abc •  0.14  0.45  double double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 7b.  74 Table  7b.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) a n d t o t a l n3 HUFAs ( h i g h l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  RC  RC  RC  WILD  WV33  COMM  mean  sem  mean  % 20:5n3  15.06-B0.46  22:5n3  4.81-B0.20  22:6n3  17.81- 0.51  Total  n3 PUFAs -  Total  n3 H U F A s  3  2  sem  % 8.36  0.24  b  b  16.70- 0.25 b  1 . 8 9 0.09 b  37.68- 0.30  sem  %  3 . 1 0 0.07  3.50- 0.42  mean  28.17 n0.38 b  7.88  b  0.09  3.28 "0.25 b  15.63  0.12  b  1.81  b  0.07  26.79 «0.26 b  1  n3 PUFAs bonds  = unsaturated  n3 f a t t y  acids with  2 t o 4 double  2  n3 HUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  5 o r more  abc  double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  B = a significant difference stocks f o r t h e parameter  (a=0.05) was f o u n d between indicated. See T a b l e 7a.  r  75 lipid, the  compared w i t h  fatty  the t o t a l  acids i n the t o t a l  HUFAs were s i g n i f i c a n t l y cultured  higher  T h e r e were no o t h e r  higher  significant  between t h e s t o c k s  BQ-WV33  eggs t h a n  20:5n3 c o n t e n t  (almost of  cultured  eggs.  t h e BQ-COMM.  revealed s i g n i f i c a n t l y  stock. more HUFAs i n  i n t h e BQ-COMM e g g s .  lipids  t h e BQ-WILD e g g s .  was s i g n i f i c a n t l y  higher  i n t h e c u l t u r e d groups significantly  The w i l d eggs o f b o t h  significantly  more 22:5n3 t h a n t h e  The 22:6n3 c o n c e n t r a t i o n i n t h e WILD and WV33  COMM eggs c o n t a i n e d  different  regardless of stock but  significantly  regardless of stock.  between s t o c k s  eggs had a  groups o f each  eggs t h a n  e g g s was n o t s i g n i f i c a n t l y  eggs, a g a i n  i n the  d i f f e r e n c e s i n n3 PUFAs o r  diet  of the total  stocks also contained  the  The BQ-WV33  s t o c k and t h e RC-WILD eggs c o n t a i n e d  more 20:5n3 t h a n  o f n3 PUFAs and  i n t h e w i l d eggs t h a n  double) i n t h e w i l d groups than  either  25-38% o f  i n t h e RC-WV33 eggs and s i g n i f i c a n t l y  more HUFAs i n t h e RC-COMM  The  The l e v e l s  c o n c e n t r a t i o n o f HUFA t h a n  HUFAs between t h e f o r m u l a t e d  the  lipid.  eggs, r e g a r d l e s s o f s t o c k .  significantly  Testing  HUFA w h i c h r e p r e s e n t e d  for the diets  less  22:6n3 t h a n  Significant  were f o u n d  t h e WILD  differences  f o r 20:5n3 i n t h e WILD  g r o u p s a n d f o r 22:5n3 i n t h e COMM and WILD g r o u p s .  L e v e l s o f t h e n6 f a t t y Tables wild  8a & 8b, were a l l s i g n i f i c a n t l y  fish  stock.  a c i d s and t h e n6 PUFAs a s shown i n  than  In both  i n either  lower  i n t h e eggs o f  group o f c u l t u r e d f i s h ,  regardless of  s t o c k s , t h e eggs f r o m t h e COMM g r o u p s had  76 Table  8a.  S e l e c t e d n6 f a t t y a c i d s a n d t o t a l n6 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n - 5 f i s h f o r each group). D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  BQ WILD mean  BQ WV33  sem  mean  sem  BQ COMM mean  sem  18:2n6  0.79- 0.05  3.43 «0.07  4.80° 0.14  20:2n6  0.00- 0.00  0.37  b  0.02  0.39  b  0.04  20:4n6  1.15-"0.03  1.60  b  0.04  1.69  b  0.04  2.34- 0.07  6.61  b  0.06  8.47°«0.22  Total  1  n6 P U F A s  1  n6 PUFAs = u n s a t u r a t e d bonds  abc •  n6 f a t t y  to  acids with  2 t o 4 double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 8 b .  77  Table  8b.  S e l e c t e d n6 f a t t y a c i d s a n d t o t a l n6 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  RC WILD mean %  RC WV33 sem  mean %  sem  RC COMM mean %  sem  18:2n6  0.85- 0.08  3.85°"0.06  4.59° 0.18  20:2n6  0.00- 0.00  0.32° 0.02  0.46° 0.04  20:4n6  1.02--0.03  1.63° 0.03  1.73° 0.04  2.19- 0.13  6.97° 0.07  8.12°«0.14  Total 1  n6 P U F A s  1  n6 PUFAs = u n s a t u r a t e d bonds  abc •  n6 f a t t y  acids with  2 t o 4 double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 8 a .  means  78  significantly acid,  higher  and h i g h e r  groups. either  l e v e l s o f 18:2n6, t h e p r e d o m i n a n t n6  total  n6 PUFAs t h a n  fatty  t h e eggs f r o m t h e WV33  T h e r e were no d i f f e r e n c e s i n 20:4n6 i n t h e eggs o f stock of c u l t u r e d f i s h .  d i f f e r e n c e s were f o u n d  Small  but s i g n i f i c a n t  between t h e two s t o c k s f o r 18:2n6  WV33 > BQ-WV33), 20:4n6  (BQ-WILD > RC-WILD) and t o t a l  (RC-  n6 PUFAs  (BQ-COMM > RC-COMM).  Leray  e t al.  (1985) f o u n d  deformities i n early  embryos when t h e b r o o d s t o c k high  n6 c o n t e n t .  cultured  fish  In t h i s  occurred  was f e d an n3 d e f i c i e n t  study  before  related  (Table 2), f e r t i l i t y  eyeing,  to the observed and s u r v i v a l  were b e t t e r i n t h e COMM t h a n although  The  t h e n3:n6 was  n9, n i l and t o t a l  d i e t with a  most o f t h e m o r t a l i t y i n t h e  h i g h e s t m o r t a l i t y r e p o r t e d by L e r a y . not d i r e c t l y  rainbow t r o u t  during  the period of  However n3:n6 r a t i o mortality.  Fecundity  t o t h e eyed s t a g e  (Table  t h e WV33 g r o u p s i n b o t h  lower i n t h e former  stocks  acids  (Tables  9b) were a l l s i g n i f i c a n t l y  l o w e r i n t h e eggs o f t h e w i l d  than  regardless of stock.  higher  than  i n t h e BQ c u l t u r e d f i s h .  m o n o u n s a t u r a t e s were l o w e r i n t h e RC-WILD f i s h cultured  fish,  RC-WV33 e g g s . diet  though not s i g n i f i c a n t l y  9a & fish  The l e v e l o f  n7 m o n o u n s a t u r a t e s i n t h e eggs o f t h e BQ-WILD f i s h significantly  4)  ( T a b l e s 6a & 6 b ) .  monounsaturated f a t t y  i n the cultured f i s h ,  was  than  was The n7 i n t h e RC  so i n t h e c a s e  of the  T h e r e were no d i f f e r e n c e s i n n5 l e v e l s among t h e  groups of e i t h e r  stock.  Significant  stock d i f f e r e n c e s  79 Table  9a.  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l a n d t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t .  mean  sem  % n5  0.42  mean  sem  % 0.03  BQ COMM  BQ WV33  BQ WILD  mean  sem  %  0.45  0.00  0.43  0.00 0.10  n7  11.65~"0.31  10.02  b  0.11  10.30  n9  21.71- 0.84  28.01  b  0.75  3 0 . 3 3 ° 0.36  0.53- 0.27  1.12  nil Total monounsaturates abc  34.31-«0.98  b  0.05  3 9 . 6 0 ° 0.77  b  1.19°  42.25  b  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 9b.  0.08  0.33 means  80 Table  9b.  Monounsaturates, r e p o r t e d as n 5 , n 7 , n 9 , n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t .  RC WILD  mean  RC WV33  sem  % n5  0.47 0.05  n7  9.97-"0.22  n9  mean  sem  %  RC COMM  mean sem %  0.44 0.01  0.43  0.02  10.05- 0.16  1 0 . 6 2 0.11  21.88- 0.77  29.04  b  0.43  2 9 . 8 7 0.28  0.52- 0.26  1.03  b  0.41  0.81- 0.19  Total m o n o u n s a t u r a t e s 32.83-"0.56  40.56  0.49  4 1 . 7 3 0.19  nil  abc  b  b  b  b  b  b  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  • = a s i g n i f i c a n t d i f f e r e n c e (<x = 0.05) was f o u n d b e t w e e n stocks f o r the parameter i n d i c a t e d . See T a b l e 9 a .  81  were f o u n d o n l y  i n t h e w i l d eggs f o r n7 f a t t y  monounsaturates  (BQ-WILD > RC-WILD i n b o t h  Differences  i n fatty  RC f i s h a r e p r o b a b l y diets  that  acids  acids  cases).  between t h e eggs o f t h e w i l d BQ a n d  a reflection  of the differences i n the  t h e s e f i s h consumed t h r o u g h o u t t h e i r  example o f t h i s  and t o t a l  i s t h e n7 c o n c e n t r a t i o n  lives.  i n the total  An  lipids  w h i c h was 11.65 i n t h e BQ-WILD eggs and 9.97 i n t h e RC-WILD eggs.  Differences  (Leray  et a i . , 1984) i n t h e m a r i n e e n v i r o n m e n t s o f t h e two  stocks  may a l s o c o n t r i b u t e  affecting  i n temperature  the level  (Hazel,  1979) and s a l i n i t y  t o d i f f e r e n c e s between t h e s t o c k s by  of unsaturation  u l t i m a t e l y a t t a i n e d by t h e  eggs.  1.2.5  The  F a t t y ,acid c o m p o s i t i o n  complete f a t t y  total  lipid  each stock  acid profiles  by d i e t .  f o rthe total  Data f o r s e l e c t e d f a t t y  a c i d s and f o r  acids  lipid,  are presented  the saturated  significantly  lower i n t h e p o l a r  in  and n3 f a t t y  r e s u l t e d i n higher  t h e c u l t u r e d eggs o f b o t h  acids  acids  eggs  than  10a & 10b) i n b o t h  n3:n6 r a t i o s  stocks.  were  were  l i p i d s of the wild (Tables  for  here.  and t h e n6 a n d n9 f a t t y  those from t h e c u l t u r e d f i s h This  fraction of  A4.3 and A4.4,  higher  stocks.  lipid  4, T a b l e s  significantly  in  lipids  of the polar  a r e l i s t e d i n Appendix  assemblages o f f a t t y  As  of the polar  i n the wild  T h e r e were no  than  significant  82 Table  10a.  S a t u r a t e d , n 3 , n6 a n d n9 f a t t y a c i d s e r i e s a n d n3:n6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM (BQ) C h i n o o k b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r each group). D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t .  BQ WILD  mean %  BQ WV33  sem  mean %  sem  BQ COMM  mean %  sem  Saturates  2 7 . 8 0 - 0.52  23.78  b  0.53  •24.95  b  0.30  n3  4 5 . 5 7 - 0.86  39.66  b  0.40  37.93  b  0.51  n6  2.41- 0.15  n9  1 3 . 7 3 - 0.74  abc  • 5 . 4 9 0.08 b  •20.68  b  0.72  7.04° 0.10 19.95  b  0.26  = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same r o w .  • = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n stocks f o r the parameter i n d i c a t e d . See T a b l e 1 0 b . BQ WILD  mean n3:n6 r a t i o "  1  BQ WV33  sem  1 8 . 9 1 1.51  mean  sem  7.22 0.07  * ANOVA was n o t c o n d u c t e d on t h e n3:n6  ratios.  BQ COMM  mean sem 5.39 0.14  83 T a b l e 10b.  S a t u r a t e d , n3, n6 a n d n9 f a t t y a c i d s e r i e s a n d n3:n6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m ROBERTSON CREEK (RC) C h i n o o k b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n=3) D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t •  RC WILD mean %  RC WV33 sem  mean %  sem  RC COMM mean %  sem  Saturates  27.61- 0.67  2 4 . 8 0 ° 0.14  B 2 5 . 8 6 ° 0.55  n3  46.81- 0.71  4 1 . 2 2 ° 0.87  3 7 . 9 9 ° 0.35  0.10  • 6.11° 0.12  6.76° 0.21  13.75- 0.55  • 1 8 . 3 7 ° 1.62  1 8 . 6 1 ° 0.34  n6  2.54-  n9  ab = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was found between means with d i f f e r e n t s u p e r s c r i p t s w i t h i n the same row. • = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was found between stocks f o r t h e parameter i n d i c a t e d . See Table 10a. RC WILD mean n3:n6 r a t i o " * ANOVA  RC WV33 sem  18.43 0.89  RC COMM  mean  sem  mean  6.76  0.08  5.62 0.15  was n o t c o n d u c t e d on t h e n3:n6  ratios.  sem  84  differences fatty  between t h e w i l d  BQ-WV33  more n6 f a t t y  a c i d s and l e s s  n9 f a t t y  11a & l i b , s i g n i f i c a n t l y h i g h e r  20:5n3, 22:5n3, n3 PUFAs and HUFAs were p r e s e n t  of  o f t h e eggs o f t h e w i l d  both s t o c k s .  acid, levels  22:6n3.  levels  found t h a t before  acids,  tenacious  rainbow t r o u t  and l i p o p r o t e i n  including  therefore  while  20:5n3, d e c l i n e d  be e s s e n t i a l  for  fatty  o f rainbow acid fatty  precursor i s  diet  for  6 months  o f 22:6n3 i n t h e  levels  of other  drastically.  the reproductive  n3 f a t t y  This fatty success of  salmonids.  A  similar  p a t t e r n o f n6 f a t t y  acid  Fremont et al. (1984)  f e d an n3 d e f i c i e n t levels  stock.  the presence of  lipid  eicosanoid  retention.  fish  i n the  i n either  importance o f t h i s  spawning m a i n t a i n e d h i g h  vitellogenin  groups,  and e s s e n t i a l  The p h y s i o l o g i c a l  by t h i s  differences  o f 22:6n3 i n t h e p o l a r starvation  i n the polar  p a t t e r n was t h e f a t t y  (1980 and 1982) r e p o r t e d  a membrane component and p o s s i b l e  reflected  than  levels of  i n the cultured  T h e r e were no s i g n i f i c a n t  muscle d u r i n g  deprivation.  may  than  o f 22:6n3 among any o f t h e d i e t  unfluctuating  as  fish  The e x c e p t i o n t o t h i s  C a s t l e d i n e and B u c k l e y  trout  acids  eggs.  shown i n T a b l e s  lipids  n3, n6 o r n9  t h a n t h e BQ-COMM eggs and t h e RC-WV33 eggs had  significantly  As  i n saturated,  a c i d s b u t t h e RC-COMM eggs had s i g n i f i c a n t l y more  saturates  the  stocks  a c i d s was s e e n i n t h e p o l a r  acid  85  Table  11a.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) a n d t o t a l n3 HUFAs ( h i g h l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  BQ WILD mean  BQ WV33 sem  %  mean  sem  %  1 4 . 9 7 - 0.45  10.70° 0.25  22:5n3  5.22--0.37  3.48° 0.04  22:6n3  24.14 0.96  Total  n3 P U F A s  Total  n3 H U F A s  1  2  1.10- 0.24 4 4 . 4 7 - 0.76  mean  sem  %  20:5n3  24.69  BQ COMM  0.33  0.44° 0.11 39.23°"0.41  10.59° 0.16 3.19° 0.19 23.59  0.73  0.57° 0.09 37.36° 0.50  1  n3 PUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  2 o r 4 double  2  n3 HUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  5 o r more  abc •  double  = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (<x = 0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e l i b .  86  Table  lib.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) a n d t o t a l n3 HUFAs ( h i g h l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; W V 3 3 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  RC WILD mean  RC WV33 sem  %  mean  sem  RC COMM mean  %  %  20:5n3  15.59- 0.76  11.05  b  0.48  22:5n3  4.42-«0.38  3.35  b  0.13  3.33  0.25  23.93  0 . 4 1 0.14  0.34  22:6n3  25.16  Total  n3 P U F A s  Total  n3 H U F A s  1  2  0.42  1.41- 0.21 45.40- 0.82  25.56  1 0 . 3 9 0.03 b  b  40.81 "0.85 b  n3 PUFAs bonds  = unsaturated  n3 f a t t y  acids with  2 or 3  2  n3 HUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids with  4 o r more  •  b  0.36 0.41  b  0.18  3 7 . 6 5 ° 0.27  1  abc  sem  double double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 1 1 a .  87  lipids  a s was s e e n i n t h e t o t a l  lower l e v e l s cultured the  o f a l l n6 f a t t y  eggs  stocks,  12a & 1 2 b ) .  higher,  20:4n6 a n d t o t a l  reflected  Within  levels  higher  level  i n the polar  a b s e n c e o f 20:2n6  the  total  acids  i n the diet  n9 f a t t y highest  acids  Between  significantly  o f 20:2n6.  higher  levels  lipids  (Tables  o f t h e w i l d eggs  and d e s a t u r a t i o n  inhibit  elongation  1980).  n3 c o n c e n t r a t i o n ,  high  10a a n d 1 0 b ) .  4) f r o m (both  both  stocks)  o f 18:2n6 t o 20:4n6 levels  o f n3 f a t t y  and d e s a t u r a t i o n  o f n6 and  I t was t h e w i l d e g g s , w i t h t h e  t h a t had t h e l o w e s t  and p o l a r  a  T h i s p a t t e r n was  n3:n6 r a t i o s  I t i s known t h a t  (Halver,  both t h e t o t a l  i t was  and u s u a l l y  acids.  ( a n d 20:3n6, s e e A p p e n d i x  suggests that elongation  level  o f 20:4n6  lipids.  s o u r c e o f t h e 20:4n6 i n a l l g r o u p s may have been d i e t a r y .  B o t h 18:2n6 and 20:4n6 were p r e s e n t and  each stock,  o f n6 f a t t y  lipid  and t h e p o l a r  was n o t o c c u r r i n g .  i n the  n6 PUFAs a n d t h e RC-WV33 eggs c o n t a i n e d  The  The  significantly  i n the w i l d than  t h e RC-COMM eggs c o n t a i n e d  significantly  in  acids  namely,  COMM eggs t h a t c o n s i s t e n t l y had t h e h i g h e s t ,  significantly  of  (Tables  lipids,  a small  diet.  amount  (0.33%) o f 18:3n6 was p r e s e n t  A d i e t a r y source could  appearance o f these f a t t y fish.  If this  physiological  therefore  acids  was t h e c a s e ,  primary precursor  i n t h e COMM and WV33 d i e t s  account  i n t h e WV33 f o r the  i n t h e eggs o f t h e c u l t u r e d  i t seems u n l i k e l y t h a t  f o r eicosanoid  synthesis  20:4n6, t h e  i n mammals, h a s much  s i g n i f i c a n c e f o r Chinook, p a r t i c u l a r l y  since the  88 Table  12a.  S e l e c t e d n6 f a t t y a c i d s a n d t o t a l n6 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m BIG QUALICUM (BQ) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r each group). D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  BQ WILD mean %  BQ WV3 3  sem  mean %  sem  BQ COMM mean %  sem  18:2n6  0.28- 0.04  1.40° 0 . 0 7  2.03° 0.04  20:2n6  0.00- 0.00  0.42° 0 . 0 2  0.45°B0.03  20:4n6  1.68- 0.04  2.43°«0.07  2.88° 0.12  2.41- 0.15  5.49°n0.08  7.04° 0.10  Total 1  n6 P U F A s  1  n6 PUFAs = u n s a t u r a t e d bonds  abc  n6 f a t t y  acids with  2 t o 4 double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  • = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 1 2 b .  means  89 Table  12b.  S e l e c t e d n6 f a t t y a c i d s a n d t o t a l n6 PUFAs ( p o l y u n s a t u r a t e d f a t t y a c i d s ) (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f eggs f r o m ROBERTSON CREEK (RC) b r o o d s t o c k on t h r e e d i e t s (n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM where n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t .  RC WILD  RC WV33  mean %  sem  mean %  18:2n6  0.34-  0.02  1 . 5 7 0.05  1.83° 0.02  20:2n6  0.00-  0.00  0.40  0.59°«0.05  20:4n6  1.56-  0.04  2.68 n0.05  2.84  2.54- 0.10  6.11 n0.12  6.76° 0.21  Total 1  n6 P U F A s  1  n6 PUFAs = u n s a t u r a t e d bonds  abc •  n6 f a t t y  sem  RC COMM  b  b  0.03  b  b  acids with  mean %  sem  b  0.02  2 t o 4 double  = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 1 2 a .  90 level  o f 2 0 : 4 n 6 was s i g n i f i c a n t l y  cultured  lower i n the w i l d than  the  eggs.  Another e x p l a n a t i o n f o r the absence and i t s p r e s e n c e  i n the  of 20:2n6 i n the w i l d  e m p l o y e d on t h e GLC i n t h e s e a n a l y s e s . (1986), t h i s  c o l u m n does n o t  satisfactorily.  Ackman  of e s s e n t i a l  A c c o r d i n g t o Ackman  s e p a r a t e 2 0 : 3 n 9 and 2 0 : 2 n 6 v e r y  (1986)  fatty  suggests that normally t h i s  a c i d s which leads to  the  In view o f t h e measures  must t a k e  i n d u c e an EFA d e f i c i e n c y i n f i s h  C a s t l e d i n e and B u c k l e y ,  1980 a n d 1 9 8 2 ) ,  it  which  researchers (eg.  i s unlikely that  f o r m a t i o n o f 2 0 : 3 n 9 was o c c u r r i n g i n t h e c u l t u r e d f i s h Rather the  fatty  acid  is  there i s a  formation of 20:3n9. i n order to  the  O n t a r i o ) column  not a problem i n n u t r i t i o n a l b i o c h e m i s t r y u n l e s s deficiency  eggs  c u l t u r e d e g g s may l i e i n t h e u s e o f  S u p e l c o w a x 10 ( S u p e l c o Canada L t d . , O a k v i l l e ,  study.  in  i d e n t i f i e d as  20:2n6,  in  the  this  r e a l l y was  20:2n6.  Monounsaturates,  p a r t i c u l a r l y 1 6 : l n 7 and 1 8 : l n 9 , a r e  c o m p o n e n t s o f TAG a n d p o l a r l i p i d m o l e c u l e s , as previously.  T h e s e two f a t t y  monoenoic f a t t y  a c i d s were t h e  mentioned  predominant  a c i d s i n b o t h t h e p o l a r and t h e t o t a l  As shown i n T a b l e s 13a & 1 3 b , n9 f a t t y  important  a c i d s were  lipids.  significantly  l o w e r i n t h e p o l a r l i p i d s o f w i l d eggs t h a n c u l t u r e d e g g s . n7 f a t t y  a c i d s were not  significantly  g r o u p s o f t h e c u l t u r e d BQ o r RC e g g s . requirement  for a specific level  different  among t h e  T h i s may i n d i c a t e a  o f n7 f a t t y  acids.  The diet  91 Table 13a.  Monounsaturates, r e p o r t e d as n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f e g g s f r o m B I G QUALICUM (BQ) b r o o d s t o c k o n t h r e e d i e t s (n = 5 f i s h f o r each g r o u p ) . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t . :  BQ WILD mean  sem  mean  0.05  0.52  %  sem  mean  0.02  0.48  0.03  5.98  0.17  %  n5  0.50  n7  6.59" 0.21  6.30" 0.27  n9  1 3 . 7 3 - 0.74  20.68 »0.72  nil  0.37  0.34  Total m o n o u n s a t u r a t e s 2 1 . 1 9 - 0.91  BQ COMM  BQ WV33  b  0.71  0.36  28.21 nQ.63 fa  sem  %  19.95° 0.26 0.37  0.04  26.78 "0.14 b  a b c = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same r o w . s = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d b e t w e e n stocks f o r t h e parameter i n d i c a t e d . See T a b l e 1 3 b .  Table  13b.  Monounsaturates, reported as n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean a n d s t a n d a r d e r r o r o f t h e m e a n s ) i n POLAR L I P I D S o f e g g s f r o m ROBERTSON CREEK ( R C ) b r o o d s t o c k o n t h r e e d i e t s ( n = 5 f i s h f o r e a c h g r o u p e x c e p t COMM w h e r e n = 3) . D i e t c o d e s : COMM = c o m m e r c i a l ; W V 3 3 = W e s t V a n c o u v e r 3 3 ; WILD = n a t u r a l diet.  RC WILD mean  RC WV33 sem  % 0.59  n7  •5.21" 0.21  0.07  n9  13.75-  0.55  nil  0.17  0.11  19.72-  0.56  abc  •  sem  %  n5  Total monounsaturates  mean  RC COMM mean  sem  %  0.48  0.01  0.50  0.04  5.75"  0.07  5.99  0.17  18.37 nl.62 b  0.23  0.06  24.82~H1.58  1 8 . 6 1 ° 0.34 0.00  0.00  2 5 . 1 0 ° • 0.24  = a significant difference (a=0.05) w a s f o u n d b e t w e e n with different s u p e r s c r i p t s w i t h i n t h e same r o w . = a s i g n i f i c a n t d i f f e r e n c e (ct = 0.05) was f o u n d between stocks f o r t h e parameter i n d i c a t e d . See T a b l e 1 3 a .  means  93 Between t h e two s t o c k s ,  where t h e r e  were  significant  differences  i n m o n o u n s a t u r a t e s , t h e BQ eggs c o n s i s t e n t l y h a d  the  concentration,  greater  WILD and BQ-WV33  i e . f o r t h e n7 s e r i e s ,  > RC-WV33; f o r t h e n9 s e r i e s ,  WV33; and f o r t o t a l  BQ-WILD > RC-  BQ-WV33  > RC-  m o n o u n s a t u r a t e s , BQ-WV33 > RC-WV33 and BQ-  COMM > RC-COMM.  1.2.6  Diets  All  diet  data  a r e presented  tag  data  f o r t h e COMM d i e t  the  WV33 d i e t  i n Apppendix  lipid  A2.3) a n d f o r t o t a l  includes  B o t h d i e t s were a n a l y s e d  lipid  feed  for dry  on a wet a n d d r y w e i g h t b a s i s fatty  m a t t e r , m o i s t u r e and l i p i d specifications  This  ( T a b l e A2.1) a n d t h e f o r m u l a t i o n o f  (Table A2.2).  matter, moisture,  2.  acids  (Table A2.4).  (Table  Dry  l e v e l s were f o u n d t o match  s t a t e d on t h e f e e d  tags  of both d i e t s  (Table  A2.1).  For  clarity,  acid  comparisons o f i n d i v i d u a l  series i n the total  presented Figures  together  with  5, 6 and 7.  and p o l a r  Parallel  f o r the wild  lipids  egg d a t a  a c i d s and f a t t y  o f t h e eggs a r e  t h e same p a r a m e t e r s  shown a t t h e b o t t o m o f e a c h f i g u r e . available  fatty  i n the diets i n  f o r the wild f i s h are  Diet data  were n o t  fish.  i The in  saturated, the t o t a l  n3, n6 a n d n9 f a t t y  and p o l a r  lipids  a c i d s e r i e s and n3:n6  ratios  o f t h e eggs o f b o t h s t o c k s  on a l l  94 diets  aredisplayed  diets  (dark bars) a r e p r e s e n t e d  saturates, in  there  (see  some s a t u r a t e s 7)  Figure  t h e major  lipids  or that saturated  n9 f a t t y  However  polar  lipids  lipids  in  lipids  The f a t t y  fish.  i npolar  of thewild  a n d n3 f a t t y  lipids  lipids.  l a r g e s t component o f i n t h e WV33  of fatty  eggs o f b o t h  i n t h e e g g s , t h e n3 f a t t y  i n thetotal  It is  o f t h e COMM e g g s .  acids  The n3 s e r i e s was a l s o d o m i n a n t  and p o l a r  a c i d 16:0  and p o l a r  and w i l d  l a r g e s t component  i n the  stocks.  2% o f  by t h e n9 s e r i e s .  acids  lipids,  i n the  and p o l a r  a c i d s were w i t h i n  i n t h e WV33 d i e t , f o l l o w e d  both t h e t o t a l  acids  t o monounsaturates  o f t h e COMM eggs a n d i n b o t h t h e t o t a l  l e v e l s of saturated  It  i n t h e COMM d i e t , were t h e  i n thetotal  lipids  present  acids.  acid i nthetotal  formed t h e l a r g e s t p r o p o r t i o n  and p o l a r  However,  acids  B o t h t h e COMM  fatty  were c o n v e r t e d  a c i d s , predominant  o f t h e WV33.  each other  f o r the  lipids.  i n saturated  fatty  Data f o r t h e  tendency f o r f a t t y  t h e y were o x i d i z e d .  COMM d i e t a n d t h e s e c o n d that  The  Except  i t was t h e n3 s e r i e s , t h e t h i r d  diet,  5.  i n TAGs a n d t o a l e s s e r e x t e n t  major s e r i e s p r e s e n t  total  also.  o f t h e eggs o f b o t h t h e c u l t u r e d  often present  the  i n Figure  i n thetotal  t h e WV33 d i e t s were h i g h  appears that  The  was a s t r o n g  t h e d i e t t o accumulate  and  was  as histograms  were t h e m a j o r  followed  and t h e s a t u r a t e s  series  by t h e n9 f a t t y i nthe polar  lipids. In t h i s  s t u d y , d i e t was a g r e a t e r  f a c t o r than stock  affecting  95  Figure 5: A c o m p a r i s o n of saturated,  n3, n6 and n9 fatty a c i d s (%) andn3.n6 ratios in  total ( T L ) and polar (PL) lipids of e g g s from c u l t u r e d  and wild (WILD) Big  Q u a l i c u m (BQ) and R o b e r t s o n Creek (RC) b r o o d s t o c k with the c o m m e r c i a l ( C O M M ) and West Vancouver  (WV33) d i e t s .  BQ Eggs - COMM Diet  RC Eggs - COMM Diet RATIO  SATURATES  n3  L i ] TL  n9  n3:n6 RATIO  RATIO  SATURATES  n3 • I COMM  L_S PL  n6 R88 TL  nS  n3:n6 RATIO  [Ml PL  RC Eggs - WV33 Diet  BQ Eggs - WV33 Diet  RATIO  RATIO  SATURATES  r>3 • • WV33  n6 ^  n9 TL  n3:n6 RATIO  SATURATES  C'_! PL  BQ Eggs - WILD Diet  n3  n6  • D WV33  KS> TL  n9 EES PL  RC Eggs - WILD Diet RATIO  I  SATURATES  1  n3  n6 TL  r  RATIO  l  n9  MB PL  n3:n6 RATIO  n3:tl6 RATIO  SATURATES  1  n3  n6 JSSiTL  EUPL  n9  i  r3:n6 RATIO  96 fatty  acid  profiles  Differences COMM and  The  i n fatty  (Figure  influence  of temperature  i n membranes  regime.  Although a wild  of f i s h  diet  would  high  of unsaturation the wild  temperatures  net pens.  their  i n the wild  fish  fish  not a  would  influence  eggs.  factor.  fatty  on t h e  During  their  have been e x p o s e d fish  in their  s t o c k matures  I s l a n d w h i l e the B i g Qualicum  and d i e t s would  t o lower  near  shore  o f f t h e west  s t o c k comes  o f G e o r g i a , i t may be e x p e c t e d  regimes  were  t o t h e same  o u t as a major  than the c u l t u r e d  t h e warmer S t r a i t temperature  A l lcultured  changes  o f HUFAs, t h e e f f e c t o f  S i n c e the Robertson Creek  c o a s t o f Vancouver into  be r u l e d  6 and 7.  levels of  was t h e r e f o r e  high levels  fatty  composition of  c o n t a i n h i g h l e v e l s o f n3  cannot  ocean m i g r a t i o n ,  acid  to alter  and were e x p o s e d  temperature levels  f o r the other  i n response t o temperature  Temperature  a c i d s and p a r t i c u l a r l y  water  between t h e  i n Figures  on t h e f a t t y  ( e g . H a z e l , 1979).  a t t h e same s i t e  temperature  true  below and i l l u s t r a t e d  been documented  reared  c o m p o s i t i o n were g r e a t e r  T h i s was a l s o  and t h e a b i l i t y  unsaturation has  5).  as d e s c r i b e d  membranes  acid  fish.  WV33 d i e t g r o u p s t h a n between t h e two s t o c k s on t h e  same d i e t , acids,  i n t h e eggs o f t h e c u l t u r e d  that  both d i f f e r .  The  most a p p a r e n t d i f f e r e n c e was t h e p r e s e n c e o f h i g h e r l e v e l s o f n3 and s a t u r a t e d in  the polar  than  fatty  lipid  a c i d s and l o w e r  fraction  are consistent  o f n9 f a t t y  o f t h e eggs o f t h e RC-WILD  i n t h e eggs o f t h e BQ-WILD f i s h  differences  levels  with t h e i r  (Figure  5).  respective  acids  fish  These temperature  97  Figure 6: A c o m p a r i s o n of s e l e c t e d n3 fatty a c i d s , n3 PUFAs and H U F A s in total ( T L ) and polar (PL) lipids of eggs from c u l t u r e d  and wild (WILD) Big Q u a l i c u m  (BQ) and R o b e r t s o n Creek (RC) b r o o d s t o c k with the c o m m e r c i a l and West Vancouver  33 (WV33) d i e t s .  BQ Eggs - COMM Diet  22:6n3 • i  22:6n3 XKK TL  COMM  i  HUFAs  RC Eggs - COMM Diet  1 n3 PUFAs  22:6n3 •  22:6n3  WV33  KSSTL  HUFAs  22:6n3  22:6n3  EfflTL  EEDPL  _••=> • ! 22:6n3  22:6n3 E53TL  n3 PUFAs  22:6n3  22:6n3 ^  EUDPL  HUFAs  HUFAs  n3 PUFAs  151 PL  RC Eggs - WV33 Diet  BQ Eggs - WILD Diet  20:6n3  5 1  ••COMM  HUPL  BQ Eggs - WV33 Diet  20:6n3  (COMM)  TL  HUFAs  EES PL  RC Eggs - WILD Diet  n3 PUFAs ^  TL  ,' J PL  n3 PUFAs  98  Figure 7: A c o m p a r i s o n of s e l e c t e d fatty a c i d s and n6 P U F A s in total ( T L ) and polar (PL) lipids of e g g s from c u l t u r e d and wild (WILD) Big Q u a l i c u m (BQ) and R o b e r t s o n Creek (RC) b r o o d s t o c k with the c o m m e r c i a l ( C O M M ) and West Vancouver  (WV33) d i e t s .  BQ Eggs - COMM Diet  RC Eggs - COMM Diet 30 26 20 %  16 10 6  18:1n9  16:1n7  22:1n11  18:2n6 } TL  20:4n6  0  n6 PUFAs  •  E l PL  U  BQ Eggs - WV33 Diet  • •  22:1n11 WV33  18:2n6  ESS! TL  20:4n6  22;1n11 COMM  18:2n6  t\V»3 TL  20:4n8  EH  n6 PUFAs  PL  RC Eggs - WV33 Diet  %  16:1n7  16:107  16  n6 PUFAs  16:1n7  [MO PL  H  BQ Eggs - WILD Diet  22:1n11 WV33  18:2n6  E33 TL  20:4n6  n8 PUFAs  EMI PL  RC Eggs - WILD Diet 30 26 20 %  16  I  10  HI 18:1n9  16:1n7  6  22:1n11  EMI TL  18:2n6  EM  PL  20:4n6  n6 PUFAs  1 18:1n9  16:1n7  22:1n11  ESS TL  18:2n6  EMI  PL  20:4n6  n6 PUFAs  99  regimes.  The two stocks were remarkably s i m i l a r i n  composition  otherwise (see a l s o F i g u r e s 6 and 7 ) .  The  major f a t t y a c i d s i n the n3 s e r i e s of both the c u l t u r e d and  w i l d eggs were 22:6n3 and 20:5n3. present  These f a t t y a c i d s were  i n the eggs i n c o n c e n t r a t i o n s  t h a t were notably  higher  than i n the d i e t s and i t i s evident t h a t they had been accumulated and conserved i n the eggs (Figure 6). f a t t y a c i d , 20:5n3 was higher  While t h e  i n both the t o t a l and p o l a r  l i p i d s o f the w i l d eggs than i n the c u l t u r e d eggs, 22:6n3 l e v e l s were remarkably s i m i l a r , p a r t i c u l a r l y i n the fraction,  polar  i n the c u l t u r e d and w i l d eggs (Figure 6).  In view of  t h e i r p h y s i o l o g i c a l s i g n i f i c a n c e as membrane components and eicosanoid precursors  and because n3 f a t t y a c i d s have e s s e n t i a l  s t a t u s f o r salmonids, i t i s reasonable t o expect c o n s i s t e n t high l e v e l s o f these HUFAs i n the egg where they would be r e q u i r e d by the growing embryo.  As shown i n Figure 7 both the COMM and WV33 d i e t s  contained  high l e v e l s o f monounsaturates, p a r t i c u l a r l y 18:ln9, 16:ln7 and 22:lnll. lipids egg  The f i r s t two are the primary monoenes i n f i s h egg  ( C h r i s t i e , 1986)  composition  and were r e f l e c t e d a c c o r d i n g l y  but 2 2 : l n l l was apparently  eggs or i t was o x i d i z e d as i t was present amounts.  i n the  not deposited i n only very  i n the  small  As p r e v i o u s l y mentioned, 18:ln9 and 16:ln7 a r e  important c o n s t i t u e n t s o f both TAGs and of many p o l a r Both were present  lipids.  i n the w i l d eggs but not i n as l a r g e amounts  100 as was found  i n the c u l t u r e d eggs.  The major n6 f a t t y a c i d present i n the COMM and WV33 d i e t s was 18:2n6 f o l l o w e d by 20:4n6.  The eggs of the c u l t u r e d f i s h  contained small amounts of these f a t t y a c i d s and the COMM eggs of both stocks contained s l i g h t l y more 18:2n6 than the WV33 eggs c o n s i s t e n t with s l i g h t l y higher l e v e l s of t h i s f a t t y i n the COMM d i e t .  acid  The low l e v e l s of 20:4n6, an important  p r o s t a g l a n d i n p r e c u r s o r f o r mammals, i n the eggs of both c u l t u r e d and w i l d f i s h  leads t o s p e c u l a t i o n t h a t t h i s  fatty  a c i d has l i t t l e p h y s i o l o g i c a l s i g n i f i c a n c e f o r Chinook.  Total  n6 PUFAs were c o n s i d e r a b l y higher i n the c u l t u r e d eggs than i n the w i l d eggs.  D i e t a r y sources of these f a t t y a c i d s are few i n  the marine environment  (Ackman, 1986).  101 1.3  CONCLUSIONS  The  i n t e n t i o n of  fatty not  acid profiles  an  objective  speculate  on  of  the  and  wild  of  n3  fatty  levels  of  n6  and  long  chain  precursors reported  has  experiment,  wild  over the  were t h a t  acids, fatty  acids.  fatty  acids  as  16-and  fed  low  levels  suggested that  of  hydroxy f a t t y  and  high  HUFAs acids  the  eggs i n t h i s  lipids  of  lower than  The  the  6,  quantitative  during  the  cultured  i n the  incubation  fish  the  leading  wild  levels  much  HUFAs, and  Leray  et  of  n6  n3:n6 r a t i o s  the higher  importance eicosanoid (1985)  as  early  as  b r o o d s t o c k were  fatty  22:6n3) a r e  Most o f  4).  lower  al.  to m o r t a l i t y  study occurred  acids. the  It  to  mortality  before eyeing  i n both t o t a l  and  was  precursors  critical  the  ( b o t h d i e t s ) were  the  in  and,  as  polar  considerably  eggs.  requirements of are  between  had  t h o u g h t t o be  recognition.  eggs  (Table  physiological  Further,  While  reproductive  lipids  wild  The  which are  cellular  shown i n T a b l e  the  and  to  broodstock  i n the  (20:5n3 and  processes of cultured  superior  s t a g e s when r a i n b o w t r o u t n3  the  Chinook salmon.  membrane components and  been d i s c u s s e d .  32-cell  composition  i t i s tempting  particularly  n9  lipid  cultured  found  embryonic d e f o r m i t i e s  the  cultured  reasons f o r the  fish  levels  for  this  t o compare  and  most o b v i o u s d i f f e r e n c e s  cultured  of  1 was  in wild  possible  performance of  The  Section  unknown.  C h i n o o k eggs f o r HUFAs The  eicosanoids  are  effective  102  in  very  low c o n c e n t r a t i o n s w h i c h s u g g e s t s  precursor  fatty  synthesis.  acids should  attributed  period,  t h e requirement  However a s work p r o c e e d s i n t h i s  eicosanoids a r e being  activated  satisfy  that a small pool of  t o them.  detected  t o maintain  has is  i n a chronic infection,  priorities  area  t o each other  fertility  the standard  and e y e i n g  t h e WV33 f i s h  i sunlikely  reservoir  Also,  little Future  diets  were  t o t h e w i l d eggs, r e g a r d l e s s o f  success  e r r o r o f t h e means,  out-performed  were a n a l y s e d f o r ( T a b l e s 2 & 4)  for reliable  relative  statistical  However t h e COMM f i s h  generally  (see a l s o Table 3 ) .  that t h e success  t h e WV33 eggs was due t o f a t t y  o f t h e COMM eggs r e l a t i v e t o acid  composition.  The n3:n6  i n t h e COMM and WV33 d i e t s were 2.73 and 5.68 ( A p p e n d i x  2, T a b l e A2.4), r e s p e c t i v e l y , COMM g r o u p t h a n lipids  Eicosanoids are  interesting.  Too few f i s h  t o be a s s e s s e d .  ratios  f o r example, a  on t h e two f o r m u l a t e d  than  significance  It  continuous  of the synthetic process.  i s bound t o be v e r y  ( F i g u r e s 5, 6 & 7 ) .  fecundity,  for a  f u n c t i o n s i s not c l e a r .  eggs o f t h e c u l t u r e d f i s h  more s i m i l a r  to  f o r other  known a b o u t t h e k i n e t i c s  stock  up and d e -  f r o m t h e membrane HUFAs and w h e t h e r t h i s  work i n t h i s  The  taken  a response  l a r g e p o o l o f p r e c u r s o r s may be n e c e s s a r y . synthesized  more  and more f u n c t i o n s a r e b e i n g  Eicosanoids are rapidly  and i n order  a s may o c c u r  area,  for their  o f both  a n d were l o w e r i n t h e eggs o f t h e  i n t h e WV33 g r o u p i n b o t h stocks  the total  and p o l a r  ( T a b l e s 6a & 6b and 10a & 1 0 b ) .  103 WV33 d i e t  The and,  was m a n u f a c t u r e d by a c o m m e r c i a l  during the year p r i o r  only twice.  Product  to this  shelf  degradation  t o v a r y i n g degrees  probably  from  this  The  dietary  life  o f 3 months  i t i s likely  critical  f o r vitamin C i n coldwater  w i t h a minimum  o f 700 mg/kg; t h e WV33 d i e t (Appendix  concluded  success.  (NRC, 1 9 8 1 ) .  1900 mg/kg  c a n be  t h a t l o s s e s o f v i t a m i n C were  to reproductive  requirement  While  o f many o f t h e n u t r i e n t s i n t h e  mg/kg d r y d i e t  with  manufactured  (NRC, 1 9 8 1 ) .  o c c u r r e d and n o t h i n g d e f i n i t i v e  study,  particularly  i t was  company  storage t h e r e f o r e exceeded the u s u a l  recommended  diet  study,  feed  The COMM d i e t was was  fish  i s 100  supplemented supplemented  2, T a b l e s A2.1 and A 2 . 2 ) .  Vitamin C  is  g e n e r a l l y added b e c a u s e t h i s v i t a m i n i s h i g h l y s u s c e p t i b l e  to  degradation  by h e a t  and m o i s t u r e  d u r i n g p r o c e s s i n g and  storage.  Despite high supplementation  prolonged  storage times  residual 1986) for  v i t a m i n C.  Slinger  resulted  et al.  i n very  (1979, c i t e d  diet, little  i n Poston,  r e p o r t l o s s e s o f 67 t o 8 3 % due t o s t o r a g e a t a b o u t  6 months.  further  Storage  stored i n black p l a s t i c  by S l i n g e r  et al.  barrels  (1979, c i t e d  i n the sun.  i n Poston,  l o s s e s o f 15 t o 6 7 % o f t h e r e m a i n i n g  activity. (C )) 2  New  a r e now  have  The f e e d f o r t h e In t h e  1986), l e e c h i n g  d u r i n g e x p o s u r e o f f e e d t o w a t e r f o r up t o 10 s e c o n d s further  20°C  c o n d i t i o n s a t t h e n e t pen s i t e may  increased the rate of degradation.  week was study  probably  o f t h e WV33  caused  vitamin C  forms o f v i t a m i n C (eg. a s c o r b a t e - 2 - s u l f a t e commonly  used  i n formulated  diets.  They a r e h e a t  104 and  water  Vitamin roles  s t a b l e a t pH 4-13 ( T u c k e r  C h a s many i m p o r t a n t  i n the prevention  manifestation cartilage); synthesis; radicals and  of s c o l i o s i s  de-toxification  compromised  synthesis  o f c o l l a g e n and  system;  4 5 0  (Sandnes, 1 9 8 8 ) .  catecholamine  i n t h e immune  system  B a s e d on t h e i r  poor  i t i s possible that  i n the f i s h  a l s o have been  and l o r d o s i s (a s c o r b u t i c  o f p o l l u t a n t s ( e g . cadmium) and f r e e  v i a t h e cytochrome P  spawning p e r f o r m a n c e ,  1986).  functions including  and amino a c i d c a t a b o l i s m ;  i n steroidogenesis  Several  biochemical  due t o i m p a i r e d  lipid  and H a l v e r ,  steroidogenesis  f e d t h e WV33 d i e t .  was  Other f u n c t i o n s  may  impaired.  studies i n various  species  (Hilton  et  ai.,1979;  Seymour, 1981 a,b; T o l b e r t , 1 9 7 9 ; Lutwak-Mann, 1 9 5 8 ; and Sandnes and B r a e k k a n , 1981 - a l l c i t e d to  a r e l a t i o n s h i p between t h e h i g h  fish  ovaries with  vitamin  C concentration  followed parallels cells  of vitamin  o f sex s t e r o i d s .  C found i n  A rise i n  i n the ovary during v i t e l l o g e n e s i s prior  to ovulation  t h e s y n t h e s i s o f sex s t e r o i d s by t h e o v a r i a n  and s u g g e s t s t h a t v i t a m i n  steroids  levels  by a d e c r e a s e i n c o n c e n t r a t i o n  hydroxylating  C may be r e q u i r e d  r e a c t i o n s necessary  such as t h e e s t r o g e n s  Waagbo et al. of  the synthesis  i n Sandnes, 1 9 8 8 ) p o i n t  to convert  (Sandnes,  in fish  f e d adequate  i nthe  c h o l e s t e r o l t o sex  1988).  (1989) demonstrated a s i g n i f i c a n t l y  17-P-estradiol  follicle  levels  higher  level,  of vitamin  C than  105 in  those fed d e f i c i e n t  vitellogenin  diets.  A higher l e v e l of c i r c u l a t i n g  i n a supplemented  g r o u p was  Sandnes et al.  (1984) found t h a t  diet  i n vitamin  deficient  tended t o produce supplemented Their  and t h e r e was  study  eggs  for fecundity a high  2).  level  survival  supplemented  Also  a commercial  significant  diet  weight  ± 9,  sd)  groups.  wet  varying  content  against  significant  data reported  hatching  with the in this (a=0.05)  i n the  group.  A  third  the experimental d i e t s ,  group.  weight  mg/kg.  associated  eggs  of s i m i l a r  ± 7,  sd) and c o m m e r c i a l  was  quality  A n a l y s i s o f t h e eggs  i n vitamin C levels  to  revealed  between t h e  s d ) , supplemented (20 y g / g wet  (31  weight  ug/g  ± 10,  of broodstock  i n m o i s t u r e , amount and c o m p o s i t i o n o f l i p i d s  (1989) r e p o r t e d  fertilization  spawning  a significantly  o f a - t o c o p h e r o l and a s c o r b i c  Eskelinen  to  1000  a study undertaken to determine the e f f e c t s  diets  broodstock fed a  s i z e were n o t  of v a r i a b i l i t y  and p r o d u c e d  differences  (15 ug/g  wet  In  egg  than i n the d e f i c i e n t  those of the supplemented  deficient  and  containing  r e p o r t e d was  g r o u p , u s e d as a c o n t r o l fed  trout  of smaller diameter than d i d  r a t e t o e y e i n g and  progeny  found.  C f o r 3 months p r i o r  the case f o r the f e c u n d i t y  (Table  greater  rainbow  broodstock fed a d i e t  figures  d a t a as was  fewer  also  to start  semi-moist f e e d which  acid  for Atlantic  salmon,  that  the highest  total  feeding  ( 9 2 . 7 % ) was  achieved with a  had  a low f a t and m o d e r a t e  and  survival  from  energy c o n t e n t  106 that  had been s u p p l e m e n t e d w i t h a s c o r b i c  The  second h i g h e s t  dry  diet  acid  rate  on s u r v i v a l  level  levels  i n the study  was a t t r i b u t a b l e  and a s c o r b i c  - 230 mg/kg.  to a-tocopherol  No  levels i n  diets.  S o l i m a n et al ( 1 9 8 6 ) d e m o n s t r a t e d t h a t 1250  a diet  supplemented t o  mg/kg s i g n i f i c a n t l y i m p r o v e d h a t c h a b i l i t y  (Oreochromis deficient  mossambicus). tilapia  finding  steroidogenesis,  impaired  affecting  exhibited  maturation  showed t h a t  vitamin C  two weeks l a t e r t h a n t h e  fed a vitamin  maturation during v i t e l l o g e n e s i s . t h e WV33 d i e t  in tilapia  Waagbo et al. ( 1 9 8 9 ) i n t e r p r e t e d  i n rainbow t r o u t ,  impaired  They a l s o  reached maturity  supplemented group.  fed  a t 1050 mg/kg.  ( 9 0 . 3 % ) was o b t a i n e d w i t h a  w h i c h had h i g h e r f a t and p r o t e i n  a t t h e second h i g h e s t  effect the  survival  acid  high  C deficient  ovarian  similar  diet, to  g r o w t h and  In the present levels  a  study,  fish  o f egg r e t e n t i o n and  (one o r b o t h o v a r i e s d i d n o t m a t u r e )  (Table  3) .  Rainbow t r o u t  feda diet  fewer  (unfertilized)  'blank'  containing this  eggs t h a n t r o u t  the rate  of f e r t i l i z a t i o n  feda diet  by H a r d y , 1 9 8 5 ) .  In  was l o w e r among t h e WV33  i n t h e COMM e g g s , a l t h o u g h t h e d i f f e r e n c e  significant fish  1400 ppm v i t a m i n C p r o d u c e d  800 ppm ( R i d e l m a n , 1 9 8 1 , c i t e d  study,  eggs t h a n  containing  was n o t  i n t h e c a s e o f t h e BQ g r o u p a n d t h e r e were t o o few  to assess  significance  i n t h e RC g r o u p  (Table 4 ) .  107 Vitamin  C i s a l s o r e q u i r e d f o r c o l l a g e n s y n t h e s i s by  embryo.  The  again  a reducing  as  f u n c t i o n of v i t a m i n agent  amino a c i d s , p r o l i n e and formation  Little  in hydroxylation  in fish  eggs b u t  i n sea  s t a r t e d a t g a s t r u l a t i o n and  d e m o n s t r a t e d an i n the  was  probably  the  embryo.  Impaired  i n c r e a s e "in the eggs o f  particularly  to  severe  performance  damage o f t h e et  al.,  slowly,  had  feed conversion  ( S o l i m a n e t al.,  From t h e  foregoing,  nutrition and  on  ovarian  growth of the  establishing  the  many t i m e s  (1988)  of  1986).  level  which  deformities, to  poor  Tilapia fry,  of vitamin  r a t e s and  in  collagen in  s p i n a l c o l u m n , and  poor  C,  grew  survival.  When  f r y performance  was  1986).  i n f l u e n c e of v i t a m i n  g r o w t h and  maturation  levels  for this  C i n broodstock  and  progeny appears c r i t i c a l .  optimum  the  after  incubation  larval  b r o o d s t o c k were f e d s u p p l e m e n t e d d i e t s ,  improved  C  Sandnes et al.  lead to  f r o m b r o o d s t o c k f e d a low  the  promoting  percentage of hydroxyproline  hatched  low  increased  increasing levels  (Soliman  The  chains.  rainbow t r o u t through  c o i n c i d e n t with  is  u r c h i n eggs c o l l a g e n  1974).  c o l l a g e n s y n t h e s i s may  hatching  hydroxylated  metabolism of v i t a m i n  b e f o r e h a t c h i n g ( G o l u b et al.,  protein  are  reactions.  o f h y d r o g e n bonds between p r o t e i n  fertilization synthesis  C in collagen synthesis  lysine,  i s known a b o u t t h e  the  vitamin  i n the  Further  survival work  in diets for  on  108 Chinook broodstock  should  be c a r r i e d o u t .  Reasons f o r t h e apparent RC-WV33 f i s h is  always  (Tables  some  success  take  facility  f o r Sea S p r i n g  Otherwise, the  environment  As  source  than  of vitamin  of these  Farm.  were t h e RC-WV33  C or t o other  effect  this  feed.  fish.  i n response t o  factors i n the diet or  fish.  i n d i c a t e d i n M a t e r i a l s and Methods, t h e r e  were 283 f i s h i n  RC-WV33 p e n a n d 1 8 9 i n t h e BQ-WV33 p e n f o r d e n s i t i e s o f c a .  the 3.41  kg/m  fish  a n d 1 4 9 RC f i s h  ca.  3  a n d 2 . 8 4 kg/m , 3  5.96 kg/m  b e l o w 6 kg/m  3  3  Groves, pers. Table  respectively.  a n d 1.05 kg/m , r e s p e c t i v e l y . 3  are considered  appropriate  f o rdensities of Pen d e n s i t i e s  i n t h e i n d u s t r y (D.  comm.) a n d a s t h e m o r p h o m e t r i c d a t a  1 indicates,  g r o w t h was n o t compromised  at the highest  that  d e n s i t y was a s i g n i f i c a n t  t h e RC-WV33  T h e r e were 811 BQ  r e a r e d on t h e COMM d i e t  fish  of  the  were more i n c l i n e d t o  t h e r e may have been some s t o c k  low l e v e l s  Salmon  g e n e r a l l y do n o t u t i l i z e  some o f t h e BQ-WV33 b r o o d s t o c k  advantage o f t h i s  There  i n t h e n e t p e n s a t Genoa Bay,  However, t h e c u l t u r e d f i s h Perhaps  over t h e  2, 3 & 4 ) c a n o n l y be g u e s s e d a t .  'wild feed'  marine broodstock  o f t h e BQ-WV33 f i s h  pen d e n s i t i e s .  broodstock.  presented i n  i n the cultured  I t i stherefore  factor  i n t h e poor  unlikely  performance  109 SECTION 2 - F e e d w i t h d r a w a l f r o m c u l t u r e d prior  2.1  to transfer  MATERIALS AND  t o f r e s h water  groups of c u l t u r e d  4-year o l d Chinook  680  m  a t Genoa Bay.  marine  t h a n 2.5  n e t pens  kg/ m  i n both pens.  3  commercial d i e t in  Section  in  1987  1.  (COMM) a t t h e same r a t e as was  by t h e b r o o d s t o c k . 1% o f body  end o f t h e summer.  The  original  i n t e n t i o n of t h i s  from the f i s h  full  month b e f o r e t r a n s f e r  spawning.  The  fish  i n Pen  i n Pen  16  usual  single  factor  fish  males.  protocol  e x p e r i m e n t was  were p r o g e n y  matured feed  of  their  or l e s s  by  t o withdraw a l l group)  f o r one  t o f r e s h water  f o r m a t u r a t i o n and  4 (full  g r o u p ) were t o a c t as  ration  f o r one week p r i o r  f o r broodstock transport.  w i t h no  fish  were  t o c a . 0.5%  (the r e s t r i c t e d  c o n t r o l s and were t o be s t a r v e d  The  Feeding l e v e l s  determine  w e i g h t p e r day a t t h e s t a r t  feed  less  f e d to the  i n t h e s e pens  to accurately  summer b u t t h e y c u t back n a t u r a l l y  the  the  Pen d e n s i t i e s were  A g a i n , not a l l of the f i s h  approximately final  salmon were h e l d i n  B o t h g r o u p s were f e d t h e same  w h i c h made i t i m p o s s i b l e  consumption  f o r maturation.  METHODS  Two  3  Chinook b r o o d s t o c k  The  to  transfer,  design  was  replication.  o f B i g Q u a l i c u m f e m a l e s and  The p u r e Chemainus s t o c k t y p i c a l l y  Chemainus  spawns b e f o r e  the  110 pure  B i g Qualicum  earlier halted  stock.  than expected. on  September  days l a t e r  and  had  Consequently, Feeding of the  2, 1987 t o be  stopped  October  fish  procedures identical  All  acid  28 and  were b r o u g h t  to the hatchery, held  fertilization  t o t h o s e employed  to  period  95%.  acid  series  fully  techniques f o r  and  lipid and  e y e i n g s u c c e s s were  i n S e c t i o n 1.  were c a r r i e d  were t r a n s f o r m e d by analyses.  The  the arc  One-way ANOVA  o u t on t h e m o r p h o m e t r i c  t o d e t e c t any  of s t a r v a t i o n .  until  of morphometric data,  i n c u b a t i o n data, composition parameters, fatty  fish  spawned a c c o r d i n g t o t h e methods  sine transformation before s t a t i s t i c a l  and  ten  t h e y were t r a n s p o r t e d on  analyses, c o l l e c t i o n  1985)  rapidly  September 16 w i t h o n l y  Sample c o l l e c t i o n ,  f o r measuring  SAS  was  Feeding of the c o n t r o l  d a t a r e p o r t e d as p e r c e n t a g e s  ( P r o c GLM,  fish  of 7 days of s a l t w a t e r s t a r v a t i o n .  i n S e c t i o n 1.  fatty  restricted  t r a n s p o r t e d on  ( c a . 2 - 4 weeks) and  outlined and  September  5 for a total  Maturing mature  on  matured  b u t t h e y were d a r k e n i n g  14 d a y s o f s a l t w a t e r s t a r v a t i o n . was  the f i s h  individual  significant  significance  level  fatty  and acids  differences was  set at  due  Ill 2.2  RESULTS AND  2.2.1  DISCUSSION  Morphometric  measurements  M o r p h o m e t r i c measurements a r e were no  significant  condition  2.2.2  The  restricted  control  fish  during  the  of  fish  the  fish  (Table  period  Fecundity,  produced 14).  not  groups of  f o r the  It i s u n l i k e l y that  significantly  longer  period  volume o r  a f f e c t e d by  smaller  fecundity  or  occurring  two  eggs no  groups  recruitment  would  i n v i t e l l o g e n e s i s (personal  observation).  major e f f e c t  egg  size  effect  No  of  rather  t h a n on  smaller  significant  egg  the egg  size  number. on  d i f f e r e n c e due  r a t e of  T h e r e was  to period  of  no  (Table  on  apparent in this  s t a r v a t i o n was  r a t e , number o f  s u r v i v a l to eyeing  have  s t a r v a t i o n was  s u r v i v a l to eyeing  between g r o u p s i n f e r t i l i z a t i o n produced, or  of  (Table  s t a r v a t i o n would  b e e n d e t e r m i n e d much e a r l i e r Therefore  eggs  smaller.  number o f  of  than  f o r the  s t a r v a t i o n as  short period  b e c a u s e egg  eggs  still  t o be  f o u n d between t h e the  There  fish.  s t a r v a t i o n , i t i s reasonable  adversely  on  14.  length, weight  I f v i t e l l o g e n e s i s was  d i f f e r e n c e was  effect  i n Table  success  w h e t h e r d e t e r m i n e d by  significant  have an  two  incubation  of  starved  spawned, was  15).  differences in fork  f a c t o r between t h e  Spawning and  presented  eyed 15).  eggs  study.  found  112 T a b l e 14.  Period of Starvation, Fork  M o r p h o m e t r i c d a t a (mean and s t a n d a r d e r r o r o f t h e means) f o r B i g Q u a l i c u m 4 - y e a r o l d b r o o d s t o c k . Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 days p r i o r t o f r e s h w a t e r t r a n s f e r .  14  days  l e n g t h , cm sem (n)  84.16 0.69 ( 5 6 )  83.58 0.74 ( 5 7 )  W e i g h t , kg sem (n)  0.19  8.44 (56)  8.41 0.21 ( 5 7 )  Condition factor sem (n)  0.10  4.69 (56)  4.78 0.12 ( 5 7 )  8.60.06 ( 1 9 )  8.3 0.11 ( 1 6 )  Egg  diameter, sem (n)  mm  to  ab = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  113  Table  15.  Period of Starvation,  Spawning and i n c u b a t i o n d a t a (mean and s t a n d a r d e r r o r o f t h e means) f o r B i g Q u a l i c u m 4 - y e a r o l d broodstock. Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 d a y s p r i o r t o f r e s h w a t e r t r a n s f e r .  '7  14  Volume o f eggs spawned, ml sem (n)  1266 52.7 (55)  1259 61.8 (57)  Fecundity (# eggs spawned) sem (n)  4702 155.3 (54)  4831 149.6 (53)  eggs, # sem (n)  4201 164.8 (53)  4474 166.5 (53)  Fertility, % sem (n)  92.8 2.90 (18)  93.8 2.39 (16)  Eyed, % sem (n)  89.05 1.91 (53)  91.96 1.41 (53)  Eyed  days  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) f o r any o f t h e spawning and i n c u b a t i o n parameters, above.  114 Ridelman rainbow  et al. ( 1 9 8 4 ) compared t h e r e p r o d u c t i v e p e r f o r m a n c e o f trout  spawning.  f e d ad libitum  No s i g n i f i c a n t d i f f e r e n c e s  characteristics, fecundity  including  days p r i o r  was i n i t i a t e d  fecundity  r=0.583, fecundity positive  (Critical  and f o r k  length  and w e i g h t  related.  fecundity  study  increases with  Significant  (ot = 0.05)  values of the c o r r e l a t i o n  correlation  group)  and between  ( r = 0 . 3 7 1 , n=60 i n t h e r e s t r i c t e d  and c o n d i t i o n  group).  factor  Correlation  revealed a small  i n the control  fish  group;  analyses of  significant  ( r = 0 . 2 7 6 , n=66) b u t a  (not s i g n i f i c a n t )  i n the r e s t r i c t e d  n=60).  ( 1 9 7 0 ) and Roy and H i g g s  (1987) r e p o r t e d t h a t  t e n d e n c y f o r egg s i z e and f e c u n d i t y In the r e s t r i c t e d f i s h  negative c o r r e l a t i o n fecundity  in this  ( r = 0 . 4 6 5 , n=60 i n t h e r e s t r i c t e d  n=66 i n t h e c o n t r o l  (r=-0.213,  a strong  so by 45  t o t h e peak o f s p a w n i n g .  n=66 i n t h e c o n t r o l  negative correlation  Blaxter  or nearly  I t was  r . Z a r , 1 9 8 4 ) f o r b o t h g r o u p s were f o u n d between  g r o u p and r = 0 . 2 8 0 , fecundity  or chemical  of starvation  i n many s p e c i e s .  correlations  coefficient,  fish  The p e r i o d  ( 1 9 7 0 ) has r e p o r t e d t h a t  positive  small  was c o m p l e t e  42 - 49 d a y s p r i o r  l e n g t h and w e i g h t  i n physical  were f o u n d i n t h e g r o u p s .  vitellogenesis  t o spawning.  f o r 45 days p r i o r t o  egg s i z e , p r o x i m a t e c o m p o s i t i o n ,  o r egg v i a b i l i t y ,  suggested that  Blaxter  or starved  t o be  there i s  inversely  t h e r e was a s i g n i f i c a n t  ( r = - 0 . 6 2 8 , n=14) between egg d i a m e t e r and  b u t t h e r e was no s i g n i f i c a n t c o r r e l a t i o n  between  115 these  parameters  Survival BQ f i s h  to eyeing  1 on t h e COMM d i e t  survival  to eyeing  were d i f f e r e n t ,  i e . BQ  Chemainus m a l e s i n S e c t i o n was p r o b a b l y in  due t o t h e i r  progeny from f o u r year  than t h a t from three Lawseth, p e r s . other  year  comm.).  abnormalities  of only  A l t h o u g h t h e two  1 and BQ c r o s s e d of the Section  age a t m a t u r i t y .  Incubation  o l d broodstock olds  (Groves, levels  i n the three  The  4) p r o d u c e d  74%.  i n Section  The h i g h  found  (Table  90%.  2  fish  success  i s generally pers.  with  greater  comm. and  o f egg r e t e n t i o n and year  old fish  f e d the  i n the four year  old fish  regardless  treatment.  2.2.3  C o m p o s i t i o n o f t h e eggs  T h e r e were no s i g n i f i c a n t matter; lipid fat plus  lipid  effects  concentration  concentration,  ash content  (Table  17).  l e s s o f each of these  level  of the t o t a l of neutral  lipid  lipids;  or p r o t e i n  (Table 16).  p o l a r and n e u t r a l  classes.  s t a r v a t i o n p e r i o d been l o n g e r ,  polar  o r on a  lipid  p e r egg were  Consistently, the r e s t r i c t e d lipid  on d r y  on a wet o r d r y w e i g h t b a s i s ;  i n t h e two g r o u p s  volumes o f t o t a l ,  calculated  due t o t h e t r e a t m e n t  as a p e r c e n t  f r e e dry weight b a s i s ;  Absolute  the  n=18).  2, t h e s u c c e s s  WV33 d i e t were n o t p r e s e n t of  (r=-0.210,  i n b o t h g r o u p s was a p p r o x i m a t e l y  of Section  progeny w i t h stocks  i n the c o n t r o l f i s h  This  greater  g r o u p had  i n d i c a t e s t h a t had effects  on  116 T a b l e 16.  Period of Starvation,  C o m p o s i t i o n o f t h e eggs (mean and s t a n d a r d e r r o r o f t h e means) o f 4 - y e a r o l d B i g Q u a l i c u m broodstock. Feed was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 days p r i o r t o f r e s h w a t e r t r a n s f e r . (n=7 f i s h f o r t h o s e s t a r v e d 7 d a y s ; n=5 f i s h f o r t h o s e s t a r v e d 14 d a y s )  days  7 mean  14 sem  mean  sem  Dry m a t t e r Moisture  40.11 59.89  0.86 0.86  39.21 60.77  0.55 0.55  Lipid, Lipid,  ww* dw"  12.84 32.01  0.35 0.40  12.85 32.78  0.19 0.48  P o l a r l i p i d , dw N e u t r a l l i p i d , dw  12.41 19.76  0.03 0.35  12.70 20.15  0.22 0.62  38.79  0.46  38.75  0.39  lipid  61.73  0.42  61.42  1.29  f a t f r e e dw  18.25  0.13  18.93  0.48  = d r y w27.27 eight  0.56  26.39  0.49  Polar of  lipid, total  Neutral of Polar of  lipid  lipid,  total lipid,  P* r owwt e i=n wet + a sw he ,i g ww h t ; dw  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e f o r any o f t h e c o m p o s i t i o n p a r a m e t e r s , above (a=0.05).  117 Table  17.  Period  A b s o l u t e volumes o f t o t a l and p o l a r l i p i d p e r egg c a l c u l a t e d f r o m mean egg volumes a n d percentages of these parameters. Eggs a r e f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  of Starvation,  Mean egg r a d i u s ,  days  7  mm  4.3 -  Mean egg volume, mm (4/3 IT r )  3  14 4.15  b  333.0  299.4  12.84  12.85  42.8  38.5  38.79  38.75  16.6  14.9  61.73  61.42  26.4  23.6  3  Lipid  concentration  A b s o l u t e volume Polar  lipid  (ww"), %  l i p i d / e g g , mm  of t o t a l  3  lipid,  A b s o l u t e volume p o l a r Neutral  lipid  %  lipid,  of t o t a l  A b s o l u t e volume n e u t r a l  mm  lipid, lipid,  3  % mm  3  ab = o r i g i n a l ANOVA r e s u l t s , s e e T a b l e 14 *ww = wet w e i g h t  118  composition period  of  may  have been f o u n d  and  t h a t even w i t h  restriction, a decline in l i p i d  this  short  concentration  was  occurring.  2.2.4  The  Fatty acid  complete  eggs o f b o t h  The  the  acid  profiles  restricted  of the  c o n t r o l eggs. i n the  fatty  No  total  shown i n T a b l e  total  other  18,  groups.  to  were v e r y  g r o u p and  4.01  T h e r e were no (Table nil  19)  f o r the  (Table  monounsaturates  acid  lipids in 4,  A4.5.  significantly  i n the  total  lipids  profile  (Table  acids  3.98  differences in  i n the  n3:n6 r a t i o s  A4.5).  total  were n o t  f o r the  lipids  subjected  restricted  d i f f e r e n c e s i n the  20)  (Table  fatty 21)  a c i d s or  i n the  total  long chain  i n the  n5,  n3  n7,  eggs f r o m t h e  total  n9  l i p i d s of the  groups of eggs.  The  than  d i f f e r e n c e s were  significant  close at  Table  the  c o n t r o l group.  significant  o r n6  14:0,  fatty  The  total  had  significant  o r n9  between t r e a t m e n t they  broodstock  fatty  lipids  i n Appendix  t h e r e were no n6  total  of the  acid,  lipid  s a t u r a t e d , n3,  ANOVA b u t  of the  groups are presented  levels  present  As  fatty  eggs f r o m t h e  higher  composition  or  two '  restricted  saturated fatty  f i s h had  a c i d s , n3  fatty  lower a b s o l u t e a c i d s , n3  HUFAs  volumes and  of  119 Table  18.  Period of Starvation,  S a t u r a t e d , n 3 , n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r to t r a n s f e r to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  days  7 mean  14 sem  mean  sem  Saturates  16.,83  0,.23  17..54  0.,36  n3  31.,91  0..57  31.,50  0..78  n6  8..02  0,.35  7..93  0..19  n9  29,.19  0,.52  28,.92  0,.19  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (ct = 0 . 0 5 ) f o r any o f t h e s e r i e s o f f a t t y a c i d s above.  Period of Starvation,  days  7  mean % n3:n6 r a t i o "  4.01  14  sem 0.21  * ANOVA was n o t c o n d u c t e d on t h e n3:n6 r a t i o s .  mean % 3.98  sem 0.18  120 Table  19.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 p o l y u n s a t u r a t e d f a t t y a c i d s (n3 PUFAs) and t o t a l h i g h l y u n s a t u r a t e d f a t t y a c i d s (HUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m 4-year o l d B i g Qualicum broodstock p r i o r t o transfer to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Period of Starvation,  7  days mean  14 sem  mean  sem  %  % 20:5n3  8.76  0.24 *  8.41  0.26  22:5n3  3.89  0.13  3.73  0.14  22:6n3  16.64  0.43  16.88  0.48  2.23  0.05  2.17  0.06  29.68  0.62  29.33  0.69  n3  PUFAs  1  n3  HUFAs  2  1  n3  PUFAs == u n s a t u r a t e d bonds  n3  fatty acids  2  n3  HUFAs  n3  fatty  = unsaturated bonds  with 2 t o 4 double  acids with  5 o r more  double  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s ( a = 0 . 0 5 ) f o r any o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  121 Table  20.  Period of Starvation,  S e l e c t e d n6 f a t t y a c i d s , t o t a l n6 p o l y u n s a t u r a t e d f a t t y a c i d s (n6 PUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Qualicum broodstock p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  7  days  ,14  mean %  sem  mean %  sem  18:2n6  4.56  0.16  4.51  0.18  20:2n6  0.42  0.02  0.40  0.02  20:4n6  1.62  0.06  1.64  0.07  8.02  0.29  7.93  0.32  n6 P U F A s 1  1  n6 PUFAs = u n s a t u r a t e d n6 f a t t y bonds  acids  with 2 t o 4 double  one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (a=0.05) f o r any o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  A  122 Table  21.  Period of Starvation,  M o n o u n s a t u r a t e s , r e p o r t e d as n 5 , n 7 , n 9 , n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  days  7 mean  14 sem  mean  sem  n5  0..46  .02  0,.44  0..01  n7  9..66  0,.12  9,.82  0..13  n9  29,.19  0,.41  28,.92  0,.46  0,.77  0,.07  0,.95  0..08  40,.08  0,.47  40,.13  0..52  nil Total  monounsaturates  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (a=0.05) f o r any o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  123 22:6n3 p e r egg ( T a b l e 22) t h a n t h e c o n t r o l indicating  2.2.5  The  that  a decline  Fatty acid  i n these f a t t y  complete f a t t y  acid  profiles  f o r the t o t a l  lipid  fatty  restricted  fish  fatty  14:0 i n t h e p o l a r  the  acid  16:0,  was a l s o  t h e s e eggs.  i n Appendix  profile,  a l s o had s i g n i f i c a n t l y  eggs o f t h e c o n t r o l  lipid  fish  higher  lipids  4, T a b l e A4.6.  7  higher  fraction  levels  of the  t h a n was p r e s e n t i n Another  i n the polar  significantly  i n the  t h e eggs o f t h e  (Table A4.6).  significantly  The o n l y o t h e r  lipids  of the polar  acid  probably  a c i d s had begun.  composition of the polar  eggs o f e a c h g r o u p a r e p r e s e n t e d  As  group,  saturate,  f r a c t i o n of fatty  acid  was 2 1 : 5 n 3 , w h i c h was l o w e r i n t h e eggs o f t h e r e s t r i c t e d  fish  t h a n i n t h e eggs o f t h e c o n t r o l  The  higher concentration  restricted  fish.  of saturates  i n t h e eggs f r o m t h e  b r o o d s t o c k was o n l y a p p a r e n t , an o f f s e t  c o n c e n t r a t i o n o f t h e n3 f a t t y in  different  t h e s e eggs  acids  (particularly  ( T a b l e s 23 and 2 4 ) . T h i s was  t o t h e lower 22:6n3) f o u n d  further  d e m o n s t r a t e d when t h e a b s o l u t e v o l u m e s o f s a t u r a t e d fatty  a c i d s p e r egg were c a l c u l a t e d  terms, t o t a l and  saturated  fatty  acids,  (Table 2 2 ) . n3 f a t t y  22:6n3 were a l l l o w e r i n t h e r e s t r i c t e d  control  group.  and n3  In a b s o l u t e  acids,  n3 HUFAs  eggs t h a n  i n the  124 22.  Table  Period Egg  A b s o l u t e v o l u m e s o f t o t a l s a t u r a t e s , n3 f a t t y a c i d s , n3 HUFAs a n d 2 2 : 6 n 3 i n t h e t o t a l a n d p o l a r l i p i d s p e r egg c a l c u l a t e d f r o m t h e a b s o l u t e v o l u m e s o f t o t a l and p o l a r l i p i d p e r egg a n d t h e p e r c e n t a g e s o f t h e s e parameters, as r e p o r t e d i n previous tables. Eggs were f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  of Starvation,  diameter,  7  days  14  8..6-  mm  3  8. 3  b  A b s o l u t e volume t o t a l  lipid,  mm  4 2 . ,8  38. 5  A b s o l u t e volume p o l a r  lipid,  mm  1 6 . ,6  14. 9  1 6 . ,83 31. ,91 29. .68 1 6 . .64  17. 31. 29. 16.  54 50 33 88  25. 39. 38. 23.  26 20 81 92  6. 12. 11. 6.  8 1 3 5  3  3  Composition of Total L i p i d : Saturates, % n3 f a t t y a c i d s , % n3 HUFAs, %  22:6n3, %  Composition of Polar Saturates, % n3 f a t t y a c i d s , % n3 HUFAs, %  Lipid:  24. .69 41, .33 40, .94 25, .10  22:6n3, %  A b s o l u t e volumes i n T o t a l L i p i d o f : Total saturates, mm T o t a l n3 f a t t y a c i d s , mm T o t a l n3 HUFAs, mm Total 22:6n3, mm 3  3  3  3  Absolute Total Total Total Total ab  volumes i n P o l a r L i p i d o f : saturates, mm n3 f a t t y a c i d s , mm n3 HUFAs, mm 22:6n3, mm  = original  3  3  3  3  ANOVA r e s u l t s ,  see T a b l e s  -  7. ,2 13, .7 12, .7 7, .1 4,.1 6,.9 6,.8 4,.2 23 a n d 24  3. 8 5. 8 5. 8 3.6  b b  125 The  n3:n6 r a t i o s were not t e s t e d by ANOVA but there was  only a  V small d i f f e r e n c e i n the n3:n6 f a t t y a c i d r a t i o s , 6.90 6.62  f o r the c o n t r o l and  There were no other groups i n t o t a l  versus  r e s t r i c t e d groups, r e s p e c t i v e l y .  s i g n i f i c a n t d i f f e r e n c e s between the  saturates,  n6 or n9 f a t t y a c i d s  two  (Table 23)  i n i n d i v i d u a l long c h a i n f a t t y a c i d s or the n3 PUFAs  or  (Table  24) . There were no s i g n i f i c a n t d i f f e r e n c e s i n i n d i v i d u a l n6 a c i d s , i n t o t a l n6 PUFAs (Table (n5,  n7,  25), or i n any  fatty  of the s e r i e s  n9, n i l ) of monounsaturates (Table 26)  i n the  two  treatment groups.  A l o s s of n3 f a t t y a c i d s i s c o n t r a r y  to the  l i t e r a t u r e which  suggests t h a t n3 f a t t y a c i d s are t e n a c i o u s l y p a r t i c u l a r l y i n p o l a r l i p i d s during and  others  are o x i d i z e d  C a s t l e d i n e and 1985a and still  B e l l et al.  r  o c c u r r i n g , the  Buckley,  Fremont et al.,  1985b).  saturates  1980;  1984;  B e l l et  f a t t y a c i d s to the eggs was  However i f v i t e l l o g e n e s i s was  i n d i c a t e t h a t t r a n s f e r of these incomplete when feed was  withdrawn.  the p e r i o d of s t a r v a t i o n been longer, more  pronounced d i f f e r e n c e s i n f a t t y a c i d composition of both t o t a l and  al.  lower l e v e l of n3 f a t t y a c i d s i n the eggs  of the r e s t r i c t e d f i s h may  Again, had  s t a r v a t i o n while  ( C a s t l e d i n e and  Buckley, 1982;  retained  p o l a r l i p i d s may  have been seen.  the  126 Table  23.  Period  of  Starvation,  S a t u r a t e d , n.3, n6 and n9 f a t t y a c i d s e r i e s and n3:n6 r a t i o s (mean and s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r to t r a n s f e r to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  days  14  7 mean  sem  mean  sem  %  %  Saturates  24 .69  0 .24  2 5 . 26  n3  41 . 3 3 -  0 .29  39. 2 0  n6  6 .05  0 .27  5. 95  0 .21  n9  18 .64  0 .57  2 0 . 23  0 .79  ab  = a  significant difference  with d i f f e r e n t  Period of Starvation,  (a = 0. 05) was f o u n d  superscripts  days  ratio"  b  0 .68  b e t w e e n means  t h e same row.  7  14  mean  sem  mean  sem  6.90  0.37  6.62  0.26  %  n3:n6  within  0 .30  * ANOVA was n o t c o n d u c t e d on t h e n 3 : n 6  ratios.  %  127  Table  24.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 p o l y u n s a t u r a t e d f a t t y a c i d s (n3 PUFAs) and t o t a l h i g h l y u n s a t u r a t e d f a t t y a c i d s (HUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f eggs f r o m 4-year o l d B i g Qualicum b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation • <n = 5 f i s h f o r e a c h <group)  P e r i o d of Starvation,  days  7  14  mean  sem  %  mean  sem  %  20:5n3  11.55  0.26  11.08  0.29  22:5n3  3.94  0.14  3.72  0.15  22:6n3  25.10  0.53  23.92  0.59  0.39  0.03  0.39  0.01  n3  PUFAs  1  n3  HUFAs  2  40.94 -  0. 44  38.81  b  0 .50  1  n3  PUFAs = u n s a t u r a t e d bonds  n3  fatty  acids with  2 to 4  2  n3 HUFAs = u n s a t u r a t e d bonds  n3  fatty  acids with  5 o r more  ab  double  double  = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  128 Table  25.  Period of Starvation,  S e l e c t e d n6 f a t t y a c i d s , t o t a l n6 p o l y u n s a t u r a t e d f a t t y a c i d s (n6 PUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d 7 o r 14 d a y s p r i o r to transfer to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  days  7  14  mean %  sem  mean %  sem  18:2n6  1.74  0.08  1.68  0.10  20:2n6  0.43  0.03  0.50  0.03  20:4n6  2.65  0.12  2.71  0.14  6.05  0.24  5.95  0.27  n6 1  PUFAs  1  n6 PUFAs = u n s a t u r a t e d bonds  n6 f a t t y  acids  with  2 to 4 double  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (a=0.05) f o r any o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  129 T a b l e 26.  M o n o u n s a t u r a t e s , r e p o r t e d as n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f eggs f r o m 4-year o l d B i g Q u a l i c u m b r o o d s t o c k s t a r v e d f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Period of S t a r v a t i o n , days  7 mean  14 sem  %  mean  sem  %  n5  0.58  0.02  0.52  0.03  n7  5.47  0.13  5.62  0.15  n9  18.64  0.63  20.22  0.71  nil  -  -  -  24.70  0.57  26.37  T o t a l monounsaturates  0.64  A one-way ANOVA c o n d u c t e d on p e r i o d o f s t a r v a t i o n i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (a=0.05) f o r a n y o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  130  2.3  CONCLUSIONS  This  s t u d y was c o n f o u n d e d  by t h e e a r l i e r  maturation of the broodstock. planned,  greater differences  profiles  and s u r v i v a l  The  s h o r t term  weight  expected  Had i t been c a r r i e d  o u t as  i n egg c o m p o s i t i o n , f a t t y  acid  r a t e s may have o c c u r r e d .  starvation  treatment  had no e f f e c t  c o n t e n t o f t h e eggs o r on t h e c o n d i t i o n broodstock.  than  However l i p i d  factor  on t h e l i p i d  of the  c o n c e n t r a t i o n (as p e r c e n t  b a s i s ) was c o n s i d e r a b l y l o w e r  lipid,  i n t h e eggs o f b o t h  wet  groups  (12.84%; 12.85%) o f f i s h compared w i t h t h e l e v e l s r e p o r t e d by (1987) f o r c u l t u r e d eggs i n 1985 (13.5-19.0%) and i n  Groves  1986 (13.5-15.5%) ( s e e A p p e n d i x 1 ) .  It  appears  starvation  from  t h i s work t h a t  o f b r o o d s t o c k on f e c u n d i t y  salmon eggs a r e n e g l i g i b l e . potential alevins  effect  and e a r l y  incorporating determine Prior  the e f f e c t s  fry.  alevin  and s u r v i v a l  Not examined  of broodstock  starvation  An e x p e r i m e n t  and e a r l y  o r d e r t o save  a farmer  f r y growth,  may w e l l  l a b o u r and f e e d  cease  costs.  i n this  term of  Chinook  s t u d y was t h e  on t h e g r o w t h o f  of longer duration,  a t what s t a g e f e e d w i t h d r a w a l  to this  of short  i s needed t o  has a n e g a t i v e  effect.  feeding broodstock i n  131 SECTION  3 - Alteration profiles  3.1  Two  MATERIALS  groups  680 m Pen  All  i n both  3  fish  4-year  transport  n e t pens a t Genoa B a y . i n Pen 11.  experiment.  starvation  diet  period,  to the industry.  i s p r e s e n t e d i n Appendix  was p r o p r i e t a r y  diet  The  but feed  2 T a b l e A2.1.  label  More  E (200 mg), more v i t a m i n C (500 mg) and l e s s c r u d e f a t  (2%) were p r e s e n t i n t h e BROOD d i e t difference  are  shown  The  fish  profiles  i n t h e COMM  Dry matter,  lipid  diet.  n o t shown on t h e f e e d  o f 4-5% o f low t e m p e r a t u r e  meal t o t h e BROOD d i e t . acid  than  between t h e two d i e t s ,  was t h e a d d i t i o n  dried  krill  c o n c e n t r a t i o n and  o f b o t h d i e t s were d e t e r m i n e d .  i n T a b l e s A2.3 and A2.4 o f A p p e n d i x  These  data  2.  were f e d a t a p p r o x i m a t e l y t h e same r a t e a s t h e f i s h  described matured  to the pre-  Pen 11 was f e d a b r o o d  information  fatty  than 3  (COMM) d u r i n g t h e y e a r  F o r s i x weeks p r i o r  o f t h e BROOD d i e t  tag,  T h e r e were 245 f i s h i n  pens.  formulation  Another  salmon were h e l d i n  Pen d e n s i t i e s were l e s s  (BROOD), c o m m e r c i a l l y a v a i l a b l e  vitamin  broodstock.  o l d Chinook  were f e d a c o m m e r c i a l  before this  Chinook  acid  AND METHODS  3 a n d 262 f i s h  kg/ m  c o m p o s i t i o n and f a t t y  i n eggs f r o m  of cultured  marine  3  of l i p i d  i n S e c t i o n s 1 a n d 2.  A g a i n , because  i n 1987, i t was i m p o s s i b l e t o c a l c u l a t e  not a l l the f i s h feed  132 consumption of the broodstock level  was  approximately  o f t h e summer b e f o r e less,  prior  Maturing  s p a w n i n g and d e c l i n e d t o c a . 0.5%,  until  fully  from  or  t h e n e t p e n s and h e l d i n  m a t u r e and spawned a c c o r d i n g t o t h e  i n S e c t i o n 1.  Sample c o l l e c t i o n  m o r p h o m e t r i c measurements, a n a l y t i c a l calculations,  those  feeding  1% o f body w e i g h t p e r day a t t h e s t a r t  were t r a n s f e r r e d  methods o u t l i n e d  measuring  However  to t r a n s p o r t to the hatchery.  fish  freshwater  accurately.  techniques  d e t e r m i n a t i o n o f egg s i z e  fertilization  and e y e i n g  procedures, and  and p r o c e d u r e s f o r  s u c c e s s were t h e same as  d e s c r i b e d i n S e c t i o n 1.  A one-way  ANOVA  ( P r o c GLM,  SAS 1985) was  m o r p h o m e t r i c and i n c u b a t i o n d a t a ; individual  fatty  a c i d s and f a t t y  significant  differences  percentages  were t r a n s f o r m e d  before analyses.  carried  composition acid  series  due t o t h e d i e t s .  o u t on t h e  parameters; to  determine  A l l data  r e p o r t e d as  by t h e a r c s i n e t r a n s f o r m a t i o n  The s i g n i f i c a n c e  level  was  and  s e t a t 95%.  133 3.2  RESULTS AND  3.2.1  Fork  M o r p h o m e t r i c measurements  l e n g t h and w e i g h t were m e a s u r e d f o r e a c h f e m a l e  and c o n d i t i o n presented in  DISCUSSION  fork  f a c t o r s were c a l c u l a t e d .  i n Table  27.  These d a t a a r e  T h e r e were no s i g n i f i c a n t  l e n g t h o r w e i g h t between t h e two d i e t  condition  factor  significantly  of the f i s h  spawned  differences  groups but t h e  on t h e BROOD d i e t was  l o w e r t h a n t h o s e on t h e COMM  diet.  Egg d i a m e t e r s were a l s o m e a s u r e d a n d no s i g n i f i c a n t d i f f e r e n c e was f o u n d between t h e two g r o u p s  3.2.2  Spawning and i n c u b a t i o n  As shown i n T a b l e significantly COMM d i e t . fecundity  28 t h e f i s h  (Table 27).  success  on t h e BROOD d i e t  produced  g r e a t e r v o l u m e s o f eggs t h a n t h o s e on t h e  However, t h e r e was no s i g n i f i c a n t  difference i n  o r egg d i a m e t e r s between t h e two g r o u p s .  o f m e a s u r i n g t h e volume o f eggs spawned i s c r u d e . fragile  The eggs a r e  a t t h e t i m e o f s p a w n i n g and h a n d l i n g must be m i n i m i z e d .  Egg v o l u m e s were m e a s u r e d by c o m p a r i s o n w i t h at  The method  100ml i n t e r v a l s on t h e s p a w n i n g b u c k e t .  difference  lines  marked o f f  Therefore the  i n egg v o l u m e s between t h e two g r o u p s was p r o b a b l y  due t o t h e i n a c c u r a c y  o f t h e method o f measurement.  T h e r e were no s i g n i f i c a n t  differences  infertilization  success,  134 Table  27.  M o r p h o m e t r i c d a t a (mean and s t a n d a r d e r r o r o f t h e means) f o r 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  COMM  Diet  BROOD  81.5 0.36 ( 1 3 6 )  81.8 0.69 ( 1 4 0 )  W e i g h t , kg sem (n)  7.6 0.07 ( 1 3 6 )  7.5 0.10 ( 1 4 0 )  Condition factor sem (n)  4.74 0.05 ( 1 3 6 )  4.49 0.05 ( 1 4 0 )  Fork  Egg  l e n g t h , cm sem (n)  diameter, sem (n)  mm  0.10  8.41 (16)  b  8.30 0.06 ( 2 9 )  ab•= a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between means w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  135  Table  28.  Spawning and i n c u b a t i o n d a t a (mean and s t a n d a r d e r r o r o f t h e means) f o r B i g Q u a l i c u m 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l d i e t (COMM) o r a b r o o d d i e t (BROOD) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation.  Diet Volume o f eggs spawned, ml sem (n)  COMM  972 21.2 (136)  BROOD  1033 22.9 (140) b  Fecundity (# eggs spawned) sem (n)  4220 130.6 (51)  4413 114.7 (67)  Eyed eggs, # sem (n)  3866 137.4 (51)  3979 118.3 (67)  Fertility, % sem (n)  94.8 2.08 (13)  93.9 3.96 (18)  Eyed, % sem (n)  91.79 1.75 (51)  91.89 1.42 (67)  ab  = a s i g n i f i c a n t d i f f e r e n c e ( a = 0 . 0 5 ) was f o u n d between means w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  136  number o f e y e d eggs p r o d u c e d o r i n t h e s u r v i v a l between t h e two g r o u p s  Correlation  and  (a=0.05;  r=0.403,  (COMM d i e t :  BROOD d i e t :  r=0.438,  no s i g n i f i c a n t  factor  r=0.099, (1987)  n=67)  related  group.  r=-0.212,  (COMM d i e t :  n=18) was f o u n d  n=51; BROOD  Significantly  r=0.250,  i n either  between egg  n=13; BROOD d i e t :  group i n t h i s  2 9 , no s i g n i f i c a n t  greater  b a s i s ) were e v i d e n t  lipid  difference  a percent of t o t a l  COMM g r o u p .  levels  study.  However,  i n dry matter  groups.  (on a wet and d r y w e i g h t  i n t h e eggs o f f i s h  t h o s e f e d t h e BROOD d i e t .  lipid  diet:  and f e c u n d i t y t o  correlation  c o n c e n t r a t i o n was f o u n d between t h e two d i e t  (as  condition  C o m p o s i t i o n o f t h e eggs  As shown i n T a b l e  in  (1970)  Roy and H i g g s  b u t no s i g n i f i c a n t  d i a m e t e r and f e c u n d i t y  3.2.3  were f o u n d between r=-0.207,  n=67)  r=0.290,  by B l a x t e r  r e p o r t e d a s t r o n g t e n d e n c y f o r egg s i z e  be i n v e r s e l y  r=0.320,  (COMM d i e t :  n=67) a s p r e d i c t e d  (COMM d i e t :  i n either  l e n g t h and  n=51; BROOD d i e t :  correlations  and f e c u n d i t y  significant  Z a r , 1984) between f o r k  between body w e i g h t and f e c u n d i t y  n=51; but  28).  a n a l y s e s o f both groups r e v e a l e d  correlations fecundity  (Table  t o eyeing  The p o l a r  f e d t h e COMM d i e t lipid  concentration  l i p i d ) was s i g n i f i c a n t l y on a f a t - f r e e  than  lower i n t h e  d r y weight b a s i s ,  c o n c e n t r a t i o n was n o t s i g n i f i c a n t l y d i f f e r e n t  the polar  between t h e  137 Table  29.  C o m p o s i t i o n o f t h e eggs (mean and s t a n d a r d e r r o r o f t h e means) o f 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l (COMM) d i e t p r i o r t o f r e s h w a t e r transfer. (n=5 f i s h f o r e a c h g r o u p )  BROOD  COMM  Diet mean  sem  mean  sem  Dry m a t t e r Moisture  41.36 58.64  0.40 0.40  40.75 59.25  Lipid, Lipid,  ww" dw"  13.58 32.89  0.14 0.65  12.39 30.29  P o l a r l i p i d , dw N e u t r a l l i p i d , dw  12.04 21.20  0.11 0.19  12.16 18.38  Polar of  lipid, total lipid  36.66  0.85  40.12  Neutral lipid, of t o t a l l i p i d  64.35  0.97  60.67  0.45  Polar of  17.99  0.43  18.35  0.40  27.76  0.54  28.36  0.33  lipid, f a t f r e e dw  Protein  ww ab  + a s h , ww wet  weight;  0.26 0 . 26 fa b  b  b  0.15 0.42 0.19 0.16 0.99  dw = d r y w e i g h t  = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d between means w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  138  two g r o u p s . the  The l o w e r p o l a r  COMM d i e t  g r o u p was t h e r e f o r e  significantly t h e s e eggs.  lipid  higher Feed  neutral  label  T a b l e A2.1, shows t h a t  free  extract  analysis  o f each d i e t  i n t h e eggs  from  to r e f l e c t the  concentration present i n  information,  presented  i n A p p e n d i x 2,  t h e COMM d i e t  contained  up t o 2% more  a n d t h e same l e v e l s  c r u d e f i b r e and a s h .  nitrogen  found  thought  lipid  c r u d e f a t t h a n t h e BROOD d i e t crude p r o t e i n ,  level  The c o n c e n t r a t i o n o f  was n o t l i s t e d . revealed  of moisture,  lipid  A single  proximate  concentrations  (wet w e i g h t  b a s i s ) o f 16.31% a n d 15.85% f o r t h e COMM a n d BROOD d i e t s , respectively. fish to  The h i g h e r  on t h e COMM d i e t  the higher  3.2.4  The  Fatty  level  acid  A4.7.  the  o f crude  acid  and lower  eggs o f f i s h  concentration  lipid  of thet o t a l  levels  levels  of 16:ln7  f e d t h e COMM d i e t  related  i nthe diet.  lipids  profiles of thetotal  higher  i n t h e eggs o f  a p p e a r s t o be d i r e c t l y  groups a r e p r e s e n t e d  Significantly  20:lnll  therefore  composition  complete f a t t y  eggs o f b o t h d i e t  lipid  lipids  i n Appendix  of 16:ln9,  i n the  4, T a b l e  18:4n3 and  a n d 2 0 : l n 9 were p r e s e n t i n than  i n t h o s e f e d t h e BROOD  diet.  No  significant  fatty 30)  acids  differences  i nthetotal  i n t h e two d i e t  statistically  i ntotal lipid  groups.  analyzed  saturates,  fatty  a c i d s were f o u n d  The n 3 : n 6  b u t were c l o s e  n 3 , n6 o r n 9  ratios  (Table  were n o t  i n v a l u e a t 3.62 a n d  139  Table  30.  S a t u r a t e d , n 3 , n6 a n d n 9 f a t t y a c i d s e r i e s a n d n 3 : n 6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k o n a b r o o d d i e t (BROOD) o r o n a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM  mean %  BROOD  sem  mean %  sem  Saturates  17.03  0.36  18.02  0.36  n3  30.44  0.99  29.59  0.20  n6  7.85  0.17  8.17  0.15  0.71  29.06  0.24  30.13  n9  A o n e - w a y ANOVA c o n d u c t e d o n d i e t i n d i c a t e d differences (a=0.05) f o r a n y o f t h e s e r i e s above.  Diet  no s i g n i f i c a n t of fatty acids  COMM mean  BROOD  sem  % n3:n6  ratio"  3.88  mean  sem  3.62  0.05  %  0.07  * ANOVA was n o t c o n d u c t e d on t h e n 3 : n 6  ratios.  140 3.88  f o r t h e BROOD and COMM eggs r e s p e c t i v e l y .  significant  T h e r e were no  d i f f e r e n c e s i n t h e n3 o r n6 PUFAs o r i n t h e n3  HUFAs i n t h e t o t a l  lipids  o f t h e two g r o u p s a s shown i n T a b l e s  31 a n d 32.  The  eggs f r o m t h e f i s h  greater those diet  levels  o f n i l monounsaturates  from f i s h contained  analyses  on t h e COMM d i e t  on t h e BROOD d i e t more 2 0 : l n l l  i n Appendix  i n the total  (Table  and 2 2 : l n l l  2, T a b l e  A2.4.  had s i g n i f i c a n t l y  33).  n9 o r t o t a l  monounsaturates.  The  complete f a t t y  eggs o f b o t h  As  composition  of the polar  profiles  groups a r e p r e s e n t e d  shown i n T a b l e  total  acid  chain on  No s i g n i f i c a n t long chain  n6 f a t t y  acids  t h e COMM d i e t  those  i n Appendix  lipids  4, T a b l e A4.8.  (Table  contained  on t h e BROOD d i e t  differencesi n  f o r t h e COMM eggs t h a n  acids  36).  (Table  35).  f o r the  i n the  35) o r i n t h e l o n g  However, t h e eggs f r o m  significantly  (Table  lipid  were n o t s u b j e c t e d t o  d i f f e r e n c e s were f o u n d  n3 f a t t y  i n the  acids i n the polar  The n3:n6 r a t i o s  ANOVA b u t i t was s l i g h t l y h i g h e r  individual  lipids  34, t h e r e were no s i g n i f i c a n t  o f t h e eggs.  BROOD e g g s .  g r o u p s i n n5, n7,  of the polar  s a t u r a t e s , n3, n6 o r n9 f a t t y  fraction  The COMM  O t h e r w i s e t h e r e were no  d i f f e r e n c e s between t h e two d i e t  Fatty acid  than  a s shown i n t h e d i e t  significant  3.2.5  lipids  fish  more n3 PUFA t h a n d i d  The f a t t y  acid  analyses of  141  Table  31.  S e l e c t e d n3 . f a t t y a c i d s , t o t a l n3 p o l y u n s a t u r a t e d f a t t y a c i d s (n3 PUFAs) and t o t a l h i g h l y u n s a t u r a t e d f a t t y a c i d s (HUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM  BROOD  mean  sem  mean  sem  20:5n3  8.12  0.45  8.29  0.15  22:5n3  3.32  0.22  3.48  0.08  22:6n3  16.38  0.33  15.58  0.14  %  %  n3  PUFAs  1  2.27  0.08  2.00  0.10  n3  HUFAs  2  28.16  0.92  27.58  0.25  1  n3  PUFAs = u n s a t u r a t e d bonds  n3  fatty  acids with  2 to 4  2  n3  HUFAs = u n s a t u r a t e d bonds  n3  fatty  acids with  5 o r more  A one-way ANOVA c o n d u c t e d on d i f f e r e n c e s (a=0.05) f o r any above.  double  double  d i e t i n d i c a t e d no s i g n i f i c a n t of the s e r i e s of f a t t y a c i d s  r  142  Table  32.  S e l e c t e d n6 f a t t y a c i d s , t o t a l n.6 p o l y u n s a t u r a t e d f a t t y a c i d s (n6 PUFAs) (mean and s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM mean  BROOD sem  Q.  mean  sem  0,  18:2n6  4.65  0.19  4.80  0.20  20:2n6  0.40  0.04  0.47  0.02  20:4n6  1.55  0.02  1.64  0.07  7.85  0.17  8.17  0.15  n6 P U F A s 1  1  n6 PUFAs = u n s a t u r a t e d bonds  n6 f a t t y  acids with  2 to 4  double  A one-way ANOVA c o n d u c t e d on d i e t i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (<x = 0.05) f o r any o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  143 Table  33.  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean a n d s t a n d a r d e r r o r o f t h e means) i n TOTAL L I P I D S o f eggs f r o m 4-year o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r a c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM  mean %  BROOD  sem  mean  sem  %  n5  0.45  0.01  0.44  0.01  n7  10.09  0.13  10.69  0.18  n9  30.13  0.71  29.06  0.24  nil Total monounsaturates  1.0241.69  0.07 0.63  0.68° 40.88  ab = a s i g n i f i c a n t d i f f e r e n c e (a=0.05) was f o u n d between w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  0.06 0.26 means  144 T a b l e 34.  S a t u r a t e d , n3, n6 a n d n9 f a t t y a c i d s e r i e s a n d n3:n6 r a t i o s (mean a n d s t a n d a r d e r r o r o f t h e means) i n POLAR L I P I D S o f e g g s f r o m 4-year o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r a c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o rmaturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM  mean  BROOD  sem  -6  mean  sem  -6  Saturates  24..68  0 .41 .  25..85  0,.54  n3  39,.87  0,.52  38,.88  0,.53  n6  6,.20  0,.14  6,.54  0,.17  n9  18,. 31  0,.35  17,.93  0,.79  A one-way ANOVA c o n d u c t e d on d i e t i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (<x = 0.05) f o r a n y o f t h e s e r i e s o f f a t t y a c i d s above.  Diet  n3:n6 r a t i o "  COMM  BROOD  mean %  sem  mean %  sem  6.45  0.21  5.95  0.12  * ANOVA was n o t c o n d u c t e d on t h e n3:n6 r a t i o s .  145 Table  35.  S e l e c t e d n3 f a t t y a c i d s , t o t a l n3 p o l y u n s a t u r a t e d f a t t y a c i d s (n3 PUFAs) and t o t a l h i g h l y u n s a t u r a t e d f a t t y a c i d s (HUFAs) (mean a n d s t a n d a r d e r r o r o f t h e mean) i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r on a r e g u l a r c o m m e r c i a l d i e t (COMM) p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  BROOD  COMM mean  sem  sem  mean  %  %  20:5n3  10.77  0.28  11.00  0.33  22:5n3  3.50  0.18  3.64  0.64  22:6n3  24.27  0.40  23.89  0.38  0.31  0.35  0.31  38.53  n3  PUFAs  1  n3  HUFAs  2  1.3338.54  b  0.04 0.53  1  n3 PUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids  with  2 t o 4 double  2  n3 HUFAs = u n s a t u r a t e d bonds  n3 f a t t y  acids  with  5 o r more  ab  double  = a s i g n i f i c a n t d i f f e r e n c e (ot = 0.05) was f o u n d between means w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  146 Table 36.  S e l e c t e d n6 f a t t y a c i d s , t o t a l n6 p o l y u n s a t u r a t e d f a t t y a c i d s (n6 PUFAs) (mean and s t a n d a r d e r r o r of t h e means) i n POLAR LIPIDS o f eggs from 4-year o l d B i g Qualicum b r o o d s t o c k on a brood d i e t (BROOD) o r on a commercial d i e t (COMM) p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f i s h f o r each group)  Diet  COMM  BROOD  mean %  sem  mean %  sem  18:2n6  2.02  0.09  1.90  0 . 05  20:2n6  0.47  0.06  0.56  0.04  20:4n6  2.57  0.05  2.67  0.09  6.20  0.14  6.54  0.17  n6 1  PUFAs  1  n6 PUFAs = u n s a t u r a t e d bonds  n6 f a t t y a c i d s w i t h 2 t o 4 d o u b l e  A one-way ANOVA c o n d u c t e d on d i e t i n d i c a t e d no s i g n i f i c a n t d i f f e r e n c e s (<x = 0.05) f o r a n y o f t h e f a t t y a c i d s o r s e r i e s o f f a t t y a c i d s above.  147 the d i e t s  (Appendix  differences were f o u n d  2, T a b l e A2.4) d i d n o t r e v e a l a n y l a r g e  i n individual i n n3 HUFAs  n3 PUFAs.  No s i g n i f i c a n t d i f f e r e n c e s  ( T a b l e 35) o r i n n6 PUFAs  ( T a b l e 36)  between eggs o f t h e two g r o u p s .  As  found  diet  contained  lipids ca.  i n the total  than  d i d those  probably  3.2.6  t h e eggs o f t h e f i s h  s i g n i f i c a n t l y more n i l f a t t y  1% more 2 2 : l n l l  that  lipids,  on t h e BROOD d i e t  i n t h e COMM d i e t  accounted  for this  on t h e COMM  acids i n the polar  (Table 37).  than  T h e r e was  i n t h e BROOD d i e t  difference  i n t h e eggs.  Diets  Feed t a g d a t a f o r t h e COMM and BROOD d i e t s a r e p r o v i d e d i n Appendix diet  2, T a b l e A2.1.  The l e v e l  o f crude  was r e p o r t e d t o be 14% (minimum) v e r s u s  t h e COMM d i e t . dried  krill  The BROOD d i e t  meal.  contained  T h i s was e x p e c t e d  composition  i n t h e eggs o f t h e f i s h  differences  i n t h e supplemented  t h e two d i e t s a l s o .  Otherwise  f a t i n t h e BROOD 16% (minimum) i n  5% low t e m p e r a t u r e  to affect on t h i s  levels  the fatty  diet.  of Vitamin  acid  T h e r e were E and C i n  t h e t a g d a t a was i d e n t i c a l  for  t h e two d i e t s .  Partial A2.3.  proximate Contrary  COMM d i e t diets.  analyses of the diets are presented  t o the feed t a g data, the moisture  was 8.50% and t h e l i p i d  levels  i n Table  level  i n the  were c a . 16% f o r  both  T h e r e f o r e t h e m a j o r d i f f e r e n c e s between t h e d i e t s  were  148 Table  37.  M o n o u n s a t u r a t e s , r e p o r t e d a s n5, n7, n9, n i l and t o t a l m o n o u n s a t u r a t e d f a t t y a c i d s (mean and s t a n d a r d e r r o r o f t h e means) i n POLAR LIPIDS o f eggs f r o m 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k on a b r o o d d i e t (BROOD) o r a c o m m e r c i a l d i e t (COMM) p r i o r to transfer to freshwater f o r maturation. (n=5 f i s h f o r e a c h g r o u p )  Diet  COMM mean  BROOD sem  mean  sem  -6  n5  0..52  0..03  0.,52  0..05  n7  5,.73  0..14  6.,07  0,.13  n9  18,.31  0,.35  17,.93  0,.79  0,.45-  nil Total  monounsaturates  25,.18  0,.07 0,.41  0,.19° 24,.79  0,.05 0,.80  ab = a s i g n i f i c a n t d i f f e r e n c e ( a = 0 . 0 5 ) was f o u n d between means w i t h a d i f f e r e n t s u p e r s c r i p t w i t h i n t h e same row.  149  the  Vitamin  the  BROOD  The  fatty  and,  for  sample fatty the  C and E l e v e l s  acid  presented  acid  and  found  in  the  krill the  might  the  BROOD d i e t  or  some  the of  acid  in  Section  1),  to  each  acids  other  in  groups  the  of  show  the  total  in  the  the  have  of  krill  A2.5.  of  profile the  meal  but,  expected  in  of  the  the  the meal  fatty  the  (krill)  seem t o do  in  reflect  not.  levels  For of  14:0  concentration levels  of  BROOD d i e t  possibility  these  that with  When c o m p a r e d the  of  than  had o x i d i z e d  acids.  A2.4  differences  others  higher  in  raises  chain  the  diets  on  sample,  krill  euphausiid  had h i g h e r  based  Table  (fed  to  COMM a n d BROOD d i e t s  were  relatively  was  the the with  broodstock similar  A2.4).  fatty  and p o l a r with  acids  lipids the  in  fatty the  lipids.  acid  eggs  of  same  The i n d i v i d u a l  and between and p o l a r  two  in  WV33 d i e t  together  diets.  the  BROOD d i e t ;  This  selected  total  Some  BROOD d i e t  been  listed a  krill  long  are  BROOD d i e t ,  constituent  similarity  themselves the  in  diets  between  COMM d i e t  (Table  eggs  respective  Table  and the  the  8 displays  the  the  COMM d i e t .  fatty  Figure  in  krill  22:6n3  acids  of  of  than  fatty  loss  of  concentration  the  16:0  20:5n3  profiles  comparison with  presence  and  presence  diet.  is  example  and the  of  and  series  the  COMM a n d t h e  parameters  graphs  of  profiles the  two  this  of  for  BROOD  the  figure  between diet  fatty  the  groups  clearly diets in  both  150  Figure 8: A comparison of selected fatty acids in the total (TL) and polar (PL) lipids of eggs of 4-year old broodstock with those in their diets (COMM and BROOD) COMM Diet  Diet  B R O O D RATIO  n3  nS  HH COMM  E33TL  SATURATES  nS  RATIO  SATURATES  n3:n6 RATIO  n3 • • BROOD  EES PI-  COMM Diet  2ftSn3  22:5n3 •JB COMM  n6  n9  ESS TL  n3m6 RATIO  HZiPL  BROOD Diet  22:6n3  n3 PUFAs  !§33 TL  E2Z1 PL  20:6n3  n3 HUFAs  22:6n3  22:6n3  •JB BROOD  M  n3 PUFAs  TL  n3 HUFAs  EM PL  BROOD Diet  COMM Diet 60  SO  4  j  1  1  16:1n7  1  18:109  1  ^  22:1n11  • • COMM  18:2n6  SS! TL  20:4(|8  [11 PL  ill n6 PUFAs  1  18:1n7  1  18:1n9  1  22:1n11  • • BROOD  ^  BSU  Bli  18:2nS  nS PUFAs  TL  20:4n6  EM PL  151 Egg  composition  Saturates diets  was n o t c o n s i s t e n t l y a r e f l e c t i o n  and n9 f a t t y  b u t t h e n3 f a t t y  concentration  a c i d s were t h e p r e d o m i n a n t a c i d s were f o u n d  i n the t o t a l  s e r i e s i n the  i n the greatest  o f b o t h g r o u p s o f eggs  n9 f a t t y a c i d s ( p r i m a r i l y 18:ln9) a c l o s e s e c o n d .  the  polar the  lipids  highest  o f b o t h g r o u p s o f eggs c o n t a i n e d concentration.  c o n t r i b u t o r s to the high fatty  The of  lipids  of the d i e t .  elongation  n3 l e v e l s  i n both the t o t a l  final  acids, particularly  and d e s a t u r a t i o n egg p r o f i l e s .  concentration  well  n3 f a t t y  acids i n  and p o l a r  acids.  essential fatty  due  The  The n3 HUFAs were t h e p r i m a r y  d i e t s were n o t t h e s o l e s o u r c e o f f a t t y  the  with  of f a t t y  acids also contributed to  lipids  and d e s a t u r a t i o n  as t o t h e p r e s e n c e o f h i g h  Retention  t h e n3 HUFAs, and  The s o u r c e o f t h e h i g h  i n t h e egg t o t a l  to elongation  acids.  a p p e a r e d t o be  of saturated  levels  18:ln9  fatty  i n the d i e t s .  partially a c i d s as  152  3.3  The  CONCLUSIONS  BROOD and COMM d i e t s  fatty  acid  composition  composition.  higher  greater  lipid  proportion  level  of neutral  broodstock f e d t h i s be  the past  eyeing.  differences  to polar diet.  may have  lipids  period of  resulted.  resulted  i n a greater  i n t h e eggs o f t h e  No a d v a n t a g e o r d i s a d v a n t a g e  to higher  lipid  The egg m o r t a l i t y  levels  during  could  the early  which has caused  concern  t y p i c a l l y o c c u r r e d between f e r t i l i z a t i o n and  T h e r e were no s i g n i f i c a n t  between t h e two g r o u p s i n t h i s first  i n egg  been f e d o v e r t h e e n t i r e  i n t h e COMM d i e t  directly attributed  incubation period. in  t o produce major d i f f e r e n c e s  Had t h e d i e t s  vitellogenesis,  The  were n o t s u f f i c i e n t l y d i f f e r e n t i n  feeding,  an e x t r a  differences  study.  store of l i p i d  i n eyeing  rate  Beyond e y e i n g  and up t o  may be u s e f u l  i f lipid  c o n c e n t r a t i o n became l i m i t i n g t o g r o w t h and m a i n t e n a n c e .  The  BROOD d i e t  theoretically  v i t a m i n E and more No e f f e c t  the  period  advantage i n f e c u n d i t y ,  Unless feeding  c a n be shown t o r e s u l t  additional  was e v i d e n t  s u c c e s s was c o n f e r r e d  BROOD d i e t .  cost  (200 I.U./kg)  (500 mg/kg) v i t a m i n C t h a n t h e COMM  of these differences  No s i g n i f i c a n t incubation  c o n t a i n e d more  of this  diet  i n this  diet.  study.  fertilization  rate or  on t h e eggs o f t h e f i s h f e d this  a diet  over a longer  i n some s i g n i f i c a n t i s not warranted.  benefit, the  153 CONCLUDING REMARKS  Since  r  1986, t h e s u r v i v a l  b r o o d s t o c k has i m p r o v e d of  o f eggs f r o m markedly.  s m o l t s has been p r o d u c e d  high  cultured  Chinook  S i n c e 1988 an a n n u a l  surplus  by t h e i n d u s t r y and a t p r e s e n t a  i n c i d e n c e o f p r e - e y e i n g egg m o r t a l i t y  appears  t o be i n t h e  past.  The  year  1987 was p i v o t a l  The  eyed  survival  rates observed  those obtained throughout broodstock in  in  studies Eyed  survivals  of the three year o l d broodstock  1986 and p r i o r  years.  appear  reflected  r e p o r t e d here, r a n g e d  of four year o l d broodstock  manufacturers Sea  i n these  the industry.  f e d t h e COMM d i e t  t h e progeny  progeny  f o r hatchery production of smolts.  from  better  success a t  Improved b r o o d s t o c k management and  spawning and i n c u b a t i o n t e c h n i q u e s may have c o n t r i b u t e d s u c c e s s a t some  Commercial  diet  formulations are closed.  Groves  (1987, unpub. r e p o r t ) s u g g e s t e d  o f s e l e n i u m , v i t a m i n E and/or  in  commercial  during workers  1986.  I t i stherefore  which m o d i f i c a t i o n s promoted  combinations t h e seven  to the  facilities.  impossible t o determine survival.  than  f o r m u l a t i o n made by  t o be t h e main r e a s o n f o r t h i s  S p r i n g Salmon Farm L t d .  c a . 74%  t o 92% i n t h e  - significantly  Changes i n d i e t  from  diets  better  that  v i t a m i n C were low  t h a t were i n u s e and a n a l y z e d  T h e s e n u t r i e n t s have been i d e n t i f i e d  ( e g . Watanabe, 1985; H a l v e r , 1985; K i n g ,  by numerous  1985; B e l l and  154 Cowey, 1985;  Soliman et al., 1986)  production of high quality eggs.  as being important  for the  Diets manufactured since  1986  contain higher levels of vitamins E and C according to feed labels.  Selenium i s not routinely l i s t e d on feed labels.  differences in fatty acid composition  The  of eggs between cultured  and wild broodstock reported herein appear to be secondary in importance for survival i n the l i g h t of the requirements for these nutrients.  155 REFERENCES  Ackman, R.G. 1986. WCOT ( c a p i l l a r y ) g a s - l i q u i d c h r o m a t o g r a p h y . i n " A n a l y s i s o f O i l s and F a t s " , ed R . J . H a m i l t o n and J.B. R o s s e l l . E l s e v i e r Applied Science Publishers. London. Ackman, R.G. and T. T a k e u c h i . 1986. C o m p a r i s o n o f f a t t y a c i d s and l i p i d s o f s m o l t i n g h a t c h e r y - f e d and w i l d A t l a n t i c salmon Salmo salar. L i p i d s 21(2 ): 117-120. A n d e r s o n , A., T. F l e t c h e r and G. S m i t h . 1979. The r e l e a s e of prostaglandin E from the s k i n of the p l a i c e , Pleuronectes platessa L. B r . J . P h a r m a c o l . 66:547-552. 2  A n d e r s o n , A., T. 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C a n .  25:2403-2418.  V e l s e n , F . P . J . 1980. E m b r y o n i c d e v e l o p m e n t i n eggs o f s o c k e y e salmon, Oncorhynchus nerka. Can. Spec. P u b l . F i s h . A q u a t . S c i . 49:19p. Waagbo, R., T. T h o r s e n and K. S a n d n e s . 1989. R o l e o f d i e t a r y a s c o r b i c a c i d i n v i t e l l o g e n e s i s i n rainbow t r o u t (Salmo gairdneri). A q u a c u l t u r e 80:301-314. W a l l a c e , R. 1978. O o c y t e g r o w t h i n nonmammalian v e r t e b r a t e s , i n "The V e r t e b r a t e O v a r y . " e d . R.E. J o n e s . Plenum. New Y o r k . Watanabe, T. 1982. L i p i d n u t r i t i o n P h y s i o l . 73B:3-15.  in fish.  Comp.  Biochem.  Watanabe, T. 1985. I m p o r t a n c e o f t h e s t u d y o f b r o o d s t o c k n u t r i t i o n f o r f u r t h e r development o f a q u a c u l t u r e . i n " N u t r i t i o n and F e e d i n g i n F i s h . " e d . C.B. Cowey, A.M. M a c k i e and J.G. B e l l . Academic P r e s s , I n c . New Y o r k . Whyte, J.N.C. 1988. F a t t y a c i d p r o f i l e s f r o m d i r e c t methanolysis of l i p i d s i n t i s s u e of cultured species. A q u a c u l t u r e 75:193-203. Wiegand, M.D. 1982. V i t e l l o g e n e s i s i n f i s h e s . P r o c . I n t . Symp. R e p r o d . P h y s i o l . F i s h . Wageningen, N e t h e r l a n d s . 2-6 A u g u s t 1982. pp 136-146. Wiegand, M.D. and D.R. I d l e r . 1982. S y n t h e s i s o f l i p i d s by t h e r a i n b o w t r o u t (Salmo gairdneri) o v a r y in vitro. Can. J . Z o o l . 60:2683-2693. W i t h l e r , R.E., W.C. C l a r k e , B.E. R i d d e l l a n d H. K r i e b e r g . 1986. F r e s h w a t e r s u r v i v a l and g r o w t h o f c h i n o o k salmon (Onchorhynchus tshawytscha) under h a t c h e r y conditions. Can. D a t a Rep. F i s h . A q u a t . S c i . 583:33 pp. Yu, T.C. and R.O. S i n n h u b e r . 1972. E f f e c t o f d i e t a r y l i n o l e n i c a c i d and d o c o s a h e x a e n o i c a c i d on g r o w t h a n d f a t t y a c i d c o m p o s i t i o n o f r a i n b o w t r o u t (Salmo g a i r d n e r i ) . L i p i d s 7:450-454. Yu, T.C. and R.O. S i n n h u b e r . 1976. Growth r e s p o n s e o f r a i n b o w t r o u t (Salmo gairdneri) t o d i e t a r y n3 and n6 f a t t y a c i d s . A q u a c u l t u r e 8:309-317.  163 Yu, T.C. and R.O. S i n n h u b e r . 1979a. E f f e c t o f d i e t a r y n3 and n6 f a t t y a c i d s on g r o w t h and f e e d c o n v e r s i o n e f f i c i e n c y o f coho salmon (Oncorhynchus kisutch). A q u a c u l t u r e 16:31-38. Yu, T.C. and R.O. S i n n h u b e r . 1981. Use o f b e e f t a l l o w a s an e n e r g y s o u r c e i n coho salmon (Oncorhynchus kitsutch) r a t i o n s . Can. J . F i s h . A q u a t . S c i . 38:367-370. Yu, T . C , R.O. S i n n h u b e r and J.D. H e n d r i c k s . 1979b. R e p r o d u c t i o n and s u r v i v a l o f r a i n b o w t r o u t (Salmo gairdneri) f e d l i n o l e n i c a c i d as t h e o n l y s o u r c e o f e s s e n t i a l f a t t y a c i d s . L i p i d s 14(6):572-575. Yu, T . C , R.O. S i n n h u b e r and G.B. Putnam. 1977. E f f e c t o f d i e t a r y l i p i d s on f a t t y a c i d c o m p o s i t i o n o f body l i p i d i n r a i n b o w t r o u t (Salmo gairdneri). L i p i d s 12(6):495-499. Zar,  J.H. 1984. " B i o s t a t i s t i c a l A n a l y s i s . " Englewood C l i f f s , N . J . 718 pp.  Prentice-Hall,  Inc.  164 APPENDIX 1 P r e l i m i n a r y analyses of s e l e c t e d n u t r i e n t parameters i n the eggs o f w i l d and c u l t u r e d C h i n o o k salmon - 1985 & 1986. (Mean or range of v a l u e s g i v e n ) 1  1985:  C u l t u r e d Eggs  % Moisture  66.1  % L i p i d , wet w e i g h t b a s i s Fatty  acids  13.5-19.0  %18:1  %18:2 %20:5  S e l e n i u m , ug/g  dry weight  V i t a m i n A, ug/g  dry weight  31.4  5.0 20.5  Wild  Eggs 57.7  8.7-13.2 21.9  0.9 32.0  1.03  1.46  4.1  7.7  1986: %  62 .0  Moisture  % Lipid % Lipid Fatty  ( e g g s ) , wet w e i g h t b a s i s ( m u s c l e ) , wet w e i g h t b a s i s  acids  %18:1  %18:2 %20: 5  S e l e n i u m , ug/g  dry weight  V i t a m i n A, ug/g Vitamin 1  E, IU/kg  from Groves,  dry weight dry weight 1987.  13 .5-15.5 2 .4-8.0 26 .4 3 .8 10 .1  57 .7 6 .0 1 .0 23 .7 1 .0 17 .4  2 .16  3 .24  3 .2  3 .9  106 .7  73 .4  165  APPENDIX 2  D i e t Data  T a b l e A2.1: Manufacturer's feed (BROOD) d i e t s .  label  data  f o r commercial  Commercial  (COMM)  (COMM) and b r o o d  Brood  (BROOl  Moisture  10.0%  10.0%  Min.  Crude P r o t e i n  47.0%  47.0%  Min.  Crude F a t  16.0%  14.0%  3.0%  3.0%  10.0%  10.0%  Max. Crude  Fibre  Maximum A s h Actual  Phosphorus  1.5%  1.5%  Actual  Calcium  2.2%  2.2%  Actual  Sodium  0.7%  0.7%  kg  10,000  10,000  "D" p e r kg  2,400  2,400  Min.  I.U. V i t a m i n "A" p e r  Min.  I.U. V i t a m i n  Min.  I.U. V i t a m i n "E" p e r  kg  300  500  Min.  I.U. V i t a m i n "C" p e r  kg  700  1,200  I n c l u d e d i n BROOD d i e t b u t n o t s t a t e d on t h e f e e d l a b e l was 5% low t e m p e r a t u r e d r i e d k r i l l m e a l .  166 APPENDIX 2  D i e t Data  T a b l e A2.2:  Formulation  o f t h e West V a n c o u v e r 33  Ingredient:  g/kg  steam d r i e d  h e r r i n g meal  550  d r i e d whey corn d i s t . blood  (WV33) d i e t .  80 dried  solubles  flour  45 50  whole f r o z e n e u p h a u s i i d s  20  freeze-dried  10  euphausiids  wheat m i d d l i n g s  81  vitamin  supplement  1  mineral  supplement  2  43 20.4  h e r r i n g o r salmon o i l (with a n t i o x i d a n t )  75  permapel (lignin ascorbic choline  sulphonate  binder)  acid  1.9  chloride  canthaxanthin  18.9  (60%)  4.7  (10% p o t e n c y )  0.4  1000.3 1 The v i t a m i n s u p p l e m e n t s u p p l i e d t h e f o l l o w i n g l e v e l s o f n u t r i e n t s / kg d i e t : v i t a m i n A a c e t a t e 15000 I.U.; cholecalcif e r o l 3000 I.U.; D L - a l p h a - t o c o p h e r o l a c e t a t e 567 I.U.; menadione (as H e t r a z e e n ) 24.8 mg; D - c a l c i u m pantothenate 182.9 mg; p y r i d o x i n e HCl 42.2 mg; r i b o f l a v i n 56.7 mg; niacin 284 mg; f o l i c a c i d 18.9 mg; t h i a m i n e m o n o n i t r a t e 38.4 mg; b i o t i n 2.84 mg; c y a n o c o b a l a m i n 0.057 mg; i n o s i t o l 378 mg 2 The m i n e r a l s u p p l e m e n t s u p p l i e d t h e f o l l o w i n g l e v e l s o f m i n e r a l s / kg d i e t : Mn (as M n S 0 * H 0) 69.2 mg; Zn (as Z n S 0 * 7 H 0 ) 45.0 mg; Co (as C o C l * 6 H 0 ) 0.94 mg; Cu (as C u S 0 * 5 H 0 ) 4.50 mg; Fe (as F e S 0 * 7 H 0 ) 47.3 mg; I (as K I 0 ) 5.1 mg 4  2  2  /  2  2  4  4  2  2  4  3  167 APPENDIX 2  Diet  Data  T a b l e A2.3: Dry m a t t e r a n d l i p i d c o m p o s i t i o n i n S e c t i o n s 1, 2 a n d 3.  of the experimental  diets  used  COMM  WV3 3  BROOD  Dry m a t t e r % Moisture %  91.50 8.50  90.33 9.67  89.91 10.09  Lipid, Lipid,  16.31 17.83  14.38 15.92  15.85 17.63  % wet w e i g h t % d r y weight  COMM = c o m m e r c i a l d i e t ( S e c t i o n s 1 & 2) WV33 = West V a n c o u v e r 33 d i e t ( S e c t i o n 1) BROOD = b r o o d d i e t ( S e c t i o n 3)  168 APPENDIX 2  D i e t Data  Table A2.4 F a t t y a c i d p r o f i l e s of d i e t s . D i e t codes : COMM = c o m m e r c i a l , WV33 = Wes Vancouver 33, BROOD = commercial brood Fatty Acid  COMM %  WV33 %  BROOD %  14:0 0 ** 14:ln9 l a ** 16:0 16:ln9 16:ln7 16:ln5 4 ** 4a ** 5 ** 6 ** 18:0 18:ln9 18:ln7 18:ln5 18:2n6 18:3n6 18:3n3 18:4n3 20: 0? 20:lnll 20:ln9 20:ln7 20:4n6 20:4n3 20:5n3 22:lnll 22:ln9 21:5n3 22:4n6? 22:5n3 22:6n3  5.37 0.87 0.34 0.26 16.75 0.00 5.75 0.00 1.36 0.22 0.41 0.50 2.76 19.40 3.09 0.35 5.03 0.00 0.99 1.03 0.26 2.70 5.78 0.28 0.44 0.29 5.32 9.58 0.65 1.56 0.57 0.62 5.15  5.59 1.75 0.51 0.00 17.00 0.24 5 . 07 0.24 1.27 0.34 0.65 0.33 3.14 17.08 2.77 0.54 3.42 0.33 0.79 1.52 0.00 4.28 2.76 0.36 0.48 0.83 6.96 5.11 0.79 4.59 1.04 1.28 8.07  6.15 0.55 0.39 0.28 18.36 0.21 6.70 0.21 1.46 0.23 0.70 0.59 2.76 19.60 3.25 0.34 5.66 0.00 0.95 1.08 0.00 2.39 5.79 0.28 0.40 0.22 5.11 8.47 0.66 1.46 0.00 0.56 4.30  total  97.68  99.13  99.11  t o t a l n3 14.96 t o t a l n6 5.47 n3:n6 r a t i o 2.73  24.04 4.23 5.68  13.68 6.06 2.26  ** = unknown/ u n i d e n t i f i e d  fatty  acids  169 APPENDIX 2  Diet  Data  T a b l e A2.5: Dry m a t t e r , l i p i d c o m p o s i t i o n e u p h a u s i i d ( k r i l l ) sample.  Fatty Acid  Krill  14: 0 0 ** 14:ln9 l a ** 16: 0 16:ln9 16:ln7 2a ** 3 ** 4 ** 4a ** 5 **  20:4n6 20:4n3 20:5n3 21:5n3 22:4n6 • 22:5n6 22:5n3 22:6n3  7.11 0.43 1.71 0.89 21.25 8.05 0.60 0.46 0.28 1.47 2.94 1.55 0.46 1.16 1.24 7.66 5.09 1.44 0.24 2.04 4.89 1.47 0.25 1.07 0.71 0.31 10.45 0.44 4.49 1.59 0.26 6.00  total  98.00  t o t a l n3 t o t a l n6 n3:n6 r a t i o  24.39 8.47 2.88  ?  * *  6 ** 18:0 18:ln9 18:ln7 18:2n6 18:3n6 18:3n3 18:4n3 20:0? 20:ln7 -? * *  **  and f a t t y a c i d  p r o f i l e of a  %  = unknown/ u n i d e n t i f i e d  Dry m a t t e r % Moisture %  78.47  Lipid, Lipid,  23.47  fatty  % wet w e i g h t % d r y weight  acids  21.54 5.06  170 APPENDIX 3 Percent L i p i d  Formulae  f o r C a l c u l a t i n g Composition Parameters.  (wet w e i g h t b a s i s ) :  [mean d r y w e i g h t o f l i p i d i n 10 ml a l i q u o t o f l i p i d e x t r a c t * (volume o f l i p i d e x t r a c t / 10 m l ) ] * 100 % = t o t a l l i p i d i n sample (total Lipid Percent (total Lipid Percent  l i p i d i n sample/ wet (wet w e i g h t b a s i s ) Lipid  w e i g h t o f sample)  % = Percent  * 100  % = Percent  (dry weight b a s i s ) :  l i p i d i n sample/ d r y w e i g h t o f sample) (dry weight b a s i s ) Polar  * 100  Lipid  of Total  Lipid:  ( t o t a l amount l i p i d i n sample/ volume o f c h l o r o f o r m l a y e r ) * volume o f a l i q u o t a p p l i e d t o s i l i c a c a r t r i d g e s = amount l i p i d in the a l i q u o t a p p l i e d to s i l i c a c a r t r i d g e s . (polar lipid  l i p i d i n a l i q u o t applied to s i l i c a i n a l i q u o t ) * 100% = P e r c e n t P o l a r  Percent Neutral  Lipid  of T o t a l  c a r t r i d g e s / amount L i p i d of T o t a l L i p i d  Lipid:  (neutral l i p i d i n aliquot applied to s i l i c a l i p i d i n a l i q u o t ) * 100% = P e r c e n t N e u t r a l Percent  Polar  Lipid  (dry weight b a s i s ) :  percent polar l i p i d of t o t a l l i p i d * percent = Percent Polar L i p i d (dry weight b a s i s ) Percent Neutral  c a r t r i d g e s / amount L i p i d of Total L i p i d  Lipid  lipid  (dry weight)  (dry weight b a s i s ) :  percent n e u t r a l l i p i d of t o t a l l i p i d * percent l i p i d weight) = percent n e u t r a l l i p i d (dry weight b a s i s ) Percent  Polar  Lipid  o f F a t F r e e Dry  (dry  Weight:  ( p o l a r l i p i d / volume a l i q u o t a p p l i e d t o s i l i c a c a r t r i d g e s ) * volume o f c h l o r o f o r m l a y e r = t o t a l p o l a r l i p i d i n whole sample t o t a l p o l a r l i p i d / (sample d r y w e i g h t - t o t a l amount l i p i d i n sample) * 100% = P e r c e n t P o l a r L i p i d o f F a t F r e e Dry W e i g h t Percent Protein  + P e r c e n t Ash  (wet w e i g h t b a s i s ) :  100 % - [% m o i s t u r e + % l i p i d ] = P e r c e n t (wet w e i g h t b a s i s )  Protein  + Percent  Ash  171 APPENDIX 4  Fatty Acid P r o f i l e s  o f Eggs  Table A4.1: F a t t y a c i d p r o f i l e s (mean a n d s t a n d a r d e r r o r o f t h e means) o f TOTAL L I P I D i n t h e eggs o f B I G QUALICUM b r o o d s t o c k o n t h r e e diets. D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t . (n = 5 f o r a l l groups) WV33 Fatty Acid 14: 0 14:ln9 ^ ** 16: 0 16:ln9 16:ln7 4 ** 5 ** 18:0 18:ln9 18:ln7 18:ln5 1  **  18:2n6 18:2n4 18:3n6 18:3n4 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 £ ** 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 y ** 22:4n6 22:5n3 22:6n3 total  mean  sem  mean  2.08 1.73 0.25 11.71 0.60 . 5.72 0.21 0.41 4.21 23.73 4.30 0.45 0.29 3.43 ~ B 0.00 0.25 0.11 0.54 0.68 0.23 0.72 1.94 0.32 0.36 a. 0.70 1.60 0.69 8.88 ^ 0.39 0.31 0.11 0.26 • 3.47 17.73 Ek Ek EL  EL  Ek  EL  Ek  EL  Ek  Ek EL  Ek  EL  Ek  Ek  0.04 0.59 0.01 0.22 0.06 0.13 0.00 0.03 0.04 0.26 0.05 0.01 0.06 0.07 0.00 0.07 0.05 0.02 0.03 0.00 0.03 0.06 0.03 0.01 0.03 0.04 0.04 0.16 0.02 0.04 0.05 0.02 0.08 0.39  WILD sem  2-  O  98.38  COMM  'o  2.39 1.11 0.25 11.90 0.57 6 .42 0.10 0.42 4.36 26.55 3.87 0.43 0.36 4.79 0.00 0.44 0.17 0.63 0.71 0.21 0.73 2.10 0.34 0.39 0.91 1.68 0.56 7.45 0.45 0.24 0.11 0.23 2.74 14.87 98.49  t>  B  A  t>  to 2L  to k>  .a. £L  to SL £L  fa 21  D B  FA  t>  0.12 0.19 0.01 0.16 0.08 0.07 0.06 0.02 0.09 0.25 0.08 0.01 0.06 0.14 0.00 0.05 0.04 0.03 0.04 0.03 0.03 0.11 0.04 0.04 0.05 0.07 0.04 0.14 0.05 0.00 0.08 0.03 0.15 0.45  mean  sem  -6  2.43 1.71 0.26 11.85 0.42 7.76 0.56 0.42 5.12 18.71 3.89 0.42 0.00 0.79 0.32 0.00 0.47 0.72 0.85 0.50 0.40 0.86 0.00 0.00 0.00 1.15 1.34 13.70 0.13 0.75 0.18 0.40 5.49 17.09  ° • °B  to c= B  B  to B  B  to to to to to e= B  B  B  B  °B  to  c  - m  SL  0.23 0.46 0.03 0.41 0.10 0 .17 0.02 0.03 0.04 0.53 0.15 0.03 0.00 0.05 0.02 0.00 0.02 0 . 05 0.05 0.05 0.19 0.05 0.00 0.00 0.00 0.02 0.04 0.36 0.08 0.36 0.07 0.12 0.23 0.74  98.66  ** = unknown/ u n i d e n t i f i e d f a t t y a c i d s a b c = a s i g n i f i c a n t d i f f e r e n c e ( a = 0.05) was f o u n d b e t w e e n means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same r o w . B = a s i g n i f i c a n t d i f f e r e n c e (<x = 0.05) was f o u n d b e t w e e n stocks f o r the f a t t y acid indicated. See T a b l e A 4 . 2 .  172 APPENDIX  4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.2: v F a t t y a c i d p r o f i l e s (mean a n d s t a n d a r d e r r o r o f t h e means) o f TOTAL L I P I D i n t h e eggs o f ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 33; WILD = n a t u r a l d i e t . (n = 5 f o r WV33 a n d WILD; n = 3 f o r COMM) Fatty Acid 14:0 14:ln9 1 ** 16: 0 16:ln9 16:ln7 4 ** 5 ** 18: 0 18:ln9 18:ln7 18:ln5  WV33 mean  sem  & o  20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 Y ** 22:4n6 22:5n3 22:6n3 total  98.51  18:2n6 18:2n4 18:3n6 18:3n4 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 f  *  ** abc  *  mean  A  SL  A  3L  A  0.06 0.53 0.01 0.22 0 .15 0.13 0.04 0.02 0.12 0.64 0.04 0.01 0.02 0.06 0.00 0.03 0.02 0.01 0.06 0.05 0.04 0.09 0.05 0.01 0.05 0.03 0.04 0.24 0.01 0.03 0.06 0.05 0 .07 0.25  mean  sem  'o  2.29 1.04 0.24 11.15 0.54 6.47 0.04 0.33 4.77 26.16 4.15 a 0.42 0.31 a. 4.59 t> 0.00 a 0.29 a 0.11 a. 0.63 0.56 a 0.25 a 0.54 2.13 fa 0.37 a 0.46 fa 0.98 a 1.73 a 0.62 a 7.88 a 0.27 a f a 0.21 0.20 0.07 a 3.28 a 15.63 H  H  98.67  WILD sem  ?. *o  2.20 2.01 0.25 12.15 0.36 5.99 B 0.06 0.40 4.34 24.96i 4.06 a 0.44 0.32 a 3.85 B 0.00 0.30 a 0.17 •zafa 0.60 0.63 a. 0.17 a 0.66 1.70 0.31 0.32 0.78 a 1.62 a 0.66 a 8.36 a. 0.37 0.33 0.27 0.10 B 3.10 a. 16 .70 a. t>  7 **  COMM  0.07 0.13 0.00 0.10 0.04 0.10 0.03 0.02 0.14 0.20 0.06 0.01 0.01 0.14 0.00 0.02 0.05 0.03 0.02 0.01 0.04 0.11 0.04 0.03 0.03 0.03 0.04 0.07 0.10 0.03 0.15 0.05 0.19 0.09  2.54 0.91 0.20 12.17 0.54 7.01 0.16 • 0.35 5.24 19.56i 2.95 B 0.47 0.00 fa 0.84 o 0.27 B 0.00 fa 0.29 fa 0.71 1.08 fa 0.45 fa 0.41 0.87 cz: 0.00 fa 0.00 cr 0.03 fa 1.02 B 1.71 *>• 15.06 B 0.11 fa 0.37 0.25 0.30 fa 4.81 B 17.81 fa b  B  B  B  FA  B  0.26 0.17 0.05 0.32 0.08 0.14 0.05 0.01 0.07 0.66 0.09 0.05 0.00 0.07 0.03 0.00 0.04 0.09 0.13 0.06 0.19 0.06 0.00 0.00 0.03 0.03 0.21 0.46 0.08 0.05 0.11 0.02 0.20 0.51  98.47  = unknown/ u n i d e n t i f i e d f a t t y a c i d s = a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row, H = a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between stocks f o r t h e f a t t y a c i d i n d i c a t e d . See T a b l e A4.1.  173 APPENDIX 4  F a t t y A c i d P r o f i l e s o f Eggs  Table A 4 . 3 : F a t t y a c i d p r o f i l e s (mean and s t a n d a r d e r r o r o f t h e means) o f POLAR LIPID i n t h e eggs o f BIG QUALICUM b r o o d s t o c k on t h r e e diets. D i e t codes: COMM = commercial; WV33 = West Vancouver 33; WILD = n a t u r a l d i e t . (n = 5 f i s h f o r a l l groups) WV33  Fatty  Acid  mean  COMM sem  9-  1.09 5.64 0.21 15.22 0.45 2.05 0.44 0.28 7.46 12.27 4.04 0.51 1.39 0.05 0.09 0.00 .0.31 2.31 0.20 0.42 0.77 2.43 0.29 10.69 0.40 0.35 0.11 0.47 3.48 24.69  total  98.13  sem  9-  o  14: 0 14:ln9 2 ** 16:0 16:ln9 16:ln7 4 ** 4a ** 18:0 18:ln9 18:ln7 18:ln5 18:2n6 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 20:ln7 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 Y ** 22:4n6 22:5n3 22:6n3  mean  WILD  St EL  EL  EL £L  EL  ~B a  a  EL  B II  0.04 0.79 0.01 0.37 0.06 0.15 0.19 0.03 0.25 0.39 0.13 0.02 0.07 0.05 0.04 0.00 0.02 0.10 0.05 0.02 0.03 0.07 0.04 0.25 0.37 0.14 0.08 0.01 0.04 0.33  '0  0.4 90 2 3 0.13 15.55 0 . 35 2.08 1.05 0.36 8.48 13.73 3.76 0.48 2.03 0.18 0.14 0.00 0.35 2.46 0.14 0.45 1.12 2.88 0.25 10.59 0 .02 0.00 0.25 0.56 3.19 23.59 98.59  EL-  m &L b  B  b  B  t> to  a  B  EL EL  to to  ZL to  0.03 0.42 0.05 0.27 0 .09 0.10 0.39 0.02 0.29 0.23 0.08 0.03 0.04 0.05 0.06 0.00 0.02 0.14 0.06 0.03 0.03 0.12 0.06 0.16 0.03 0.00 0.12 0.02 0.19 0.73  mean 9"6  1.32 2.44 0.23 16.41 0.39 2.87 0.22 0.24 10.07 9.93 3.72 0.49 0.28 0.25 0.19 0.20 0.12 0.96 0.00 0.00 0.00 1.68 0.66 14.97 0.25 0.14 0.29 0.45 5.22 24.14  sem to 0 . 1 0  b EL  B  C3 O •  B  b  to to to to  c= C 3 to to  B  fa  0.69 0.06 0.38 0.06 0.19 0.17 0.06 0.31 0.38 0.19 0.05 0.04 0.07 0.10 0.05 0.09 0.15 0.00 0.00 0.00 0.04 0.09 0.45 0.25 0.07 0.13 0.09 0.37 0.96  98.11  ** = unknown/ u n i d e n t i f i e d f a t t y a c i d s abc = a s i g n i f i c a n t d i f f e r e n c e (a = 0 . 0 5 ) was found between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. • = a s i g n i f i c a n t d i f f e r e n c e (a = 0 . 0 5 ) was found between s t o c k s f o r t h e f a t t y a c i d i n d i c a t e d . See T a b l e A4.4.  174 APPENDIX 4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.4: F a t t y a c i d p r o f i l e s (mean a n d s t a n d a r d e r r o r o f t h e means) o f POLAR L I P I D i n t h e eggs o f ROBERTSON CREEK b r o o d s t o c k on t h r e e d i e t s . D i e t c o d e s : COMM = c o m m e r c i a l ; WV33 = West V a n c o u v e r 3 3 ; WILD = n a t u r a l d i e t . (n = 5 f o r WV33 and WILD; n = 3 f o r COMM) WV33 Fatty Acid  mean  COMM sem  3^>  14: 0 14:ln9 ^ ** 16:0 16:ln9 16:ln7 4 ** 4a * * 18:0 18:ln9 18:ln7 18:ln5 18:2n6 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 20:ln7 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 Y ** 22:4n6 22:5n3 22:6n3  0.99 3.83 0.18 15.73 0 . 32 S L 1.94 0.21 0.17 8.08 1 2 . 2 6 a. 3 . 7 6 Sk 0.47 1.57 0.02 0.09 0.00 0 . 2 3 SL 1.96 0.04 • 0.40 0.90 **• 2.68 »• 0.30 A. 11.05 0.00 0.85 a 0.00 0.55 3.35 25.56  total  97.49  ** abc  B  0.06 1.66 0.06 0.25 0.12 0.08 0.13 0.07 0.27 0.22 0.07 0.01 0.05 0.02 0.05 0.00 0.06 0.12 0.04 0.03 0.05 0.05 0.08 0.48 0.00 0.20 0.00 0.09 0.13 0.25  mean 9"o  1.09 2.30 0.24 15.37 0.00 2.18 0.22 0.08 9.39 13.48 3.73 0.50 1.83 0.07 0.00 0.00 0.00 2.83 0.08 0.59 1.10 2.84 0.27 10.39 0.00 0.00 0.33 0.41 3.33 23.93 96.77  WILD sem  0.06 0.35 0.01 0.49 *=>• 0.00 iS.t> 0.12 • 0.22 0.08 0.27 fa 0 . 4 8 0.28 0.04 fa 0 . 0 2 0.07 0.00 0.00 0.00 0.21 0.08 0.04 to 0 . 0 7 0.02 Si 0.14 0.03 0.00 fa 0 . 0 0 to 0 . 0 7 0.09 0.36 0.41  mean Q. -6  1.33 2.23 0.24 16.28 0.21 2.43 0.05 0.26 9.99 10.25 2.77 0.59 0.34 0.17 0.31 0.12 0.17 1.06 0.00 0.00 0.00 1.55 0.92 15.59 0.00 0.23 0.38 0.64 4.41 25.16  sem to  0.09 0.50 0 . 01 0.64 0 . 07 B 0.12 0.03 0.04 to 0 . 1 3 C2H 0 . 3 2 0.12 0.07 c= 0.02 0.06 0.05 B 0.06 0.11 <Z5 0 . 1 2 0.00 C 2 0.00 CJ 0.00 to 0 . 04 to 0 . 1 1 to 0 . 7 6 0.00 c: 0.03 to 0 . 0 9 0.05 B 0.38 0.42  b  B  b  B  97.67  = unknown/ u n i d e n t i f i e d f a t t y a c i d s = a s i g n i f i c a n t d i f f e r e n c e (a = 0 . 0 5 ) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row. • = a s i g n i f i c a n t d i f f e r e n c e (a = 0 . 0 5 ) was f o u n d between stocks f o r the f a t t y a c i d i n d i c a t e d . See T a b l e A 4 . 3 .  175 APPENDIX 4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.5: F a t t y a c i d p r o f i l e s (mean and s t a n d a r d e r r o r o f t h e means) o f TOTAL LIPIDS i n t h e eggs o f 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k . F e e d was w i t h d r a w n f r o m t h e s e f i s h f o r 7 d a y s o r 14 d a y s p r i o r to t r a n s f e r t o freshwater f o r maturation. (n=5 f o r t h o s e s t a r v e d 7 d a y s ; n=7 f o r t h o s e s t a r v e d 14 d a y s )  Period of S t a r v a t i o n , days Fatty Acid 14: 0 14:ln9  7 mean %  16: 0 16:ln9 16:ln7 5 ** 18:0 18:ln9 18:ln7 18:ln5 7 ** 18:2n6 18:3n6 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 22:4n6 22:5n3 22:6n3  1.70 0.37 0.19 10.65 0.77 6.00 0.38 4.48 26.67 3.66 0.46 0.34 4.56 0.36 0.68 0.77 0.20 0.53 1.38 0.42 0.84 1.62 0.78 8.76 0.24 0.38 0.23 3.89 16.64  Total  98.04  ^  **  14 sem  sem  %  0 . 04 0.05 0.05 0.13 0.05 0.20 0.10 0.10 0.55 0.11 0.02 0.03 0.16 0.03 0.02 0.04 0.02 0.03 0.07 0.02 0.06 0.06 0.05 0.24 0.06 0.16 0.11 0.13 0.43  ** = unknown/ u n i d e n t i f i e d f a t t y  ab  mean 1.95 0.53 0.26 11.27 0.71 6.10 0.44 4.33 26.08 3.72 0.44 0.35 4.51 0.40 0.66 0.78 0.23 0.61 1.60  0.40  0.78 1.64 0.73 8.41 0.34 0.30 0.19 3.73 16.88  b  0.06 0.08 0.00 0.31 0.03 0.04 0.02 0.04 0.18 0.10 0.01 0.02 0.18 0.03 0.01 0.03 0.01 0.04 0.08 0.02 0.05 0.07 0.05 0.26 0.03 0.05 0.04 0.14 0.48  98.37 acids  = a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  176 APPENDIX 4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.6: F a t t y a c i d p r o f i l e s (mean and s t a n d a r d e r r o r o f t h e means) o f POLAR LIPIDS i n t h e eggs o f 4 - y e a r o l d B i g Q u a l i c u m b r o o d s t o c k . F e e d was w i t h d r a w n f r o m t h e s e f i s h f o r 7 o r 14 d a y s p r i o r t o t r a n s f e r t o freshwater f o r maturation. (n=5 f o r t h o s e s t a r v e d 7 d a y s ; n=7 f o r t h o s e s t a r v e d 14 d a y s ) Period of Starvation,  7  days  Fatty Acid  mean  14  sem  mean  sem  % 14: 0 14:ln9 ^  **  16: 0 16:ln9 16:ln7 4 ** 4a * * 18: 0 18:ln9 18:ln7 18:ln5 18:2n6 20:ln9 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 21:5n3 22:4n6 22:5n3 22:6n3 Total ** ab  .  0.88 3.08 0.29 14.79 0.50 1.80 0.31 0.24 9.02 13.38 3.67 0.58 1.74 1.68 0.43 0.85 2.65 0.39 11.55 0.34 0.38 3.94 25.10 97.57  -  -  -  0.01 0.64 0.01 0.15 0.04 0.08 0.11 0.06 0.18 0.14 0.08 0.02 0.08 0.09 0.03 0.06 0.12 0.03 0.26 0.05 0.06 0.14 0.53  1.06 4.51 0.34 15.72 0.35 1.93 0.41 0.29 8.48 13.27 3.69 0.52 1.68 2.09 0.50 0.82 2.71 0.39 11.08 0.08 0.25 3.72 23.92  b  fa  b  0.03 0.92 0.04 0.28 0.14 0.10 0.14 0.03 0.25 0.05 0.14 0.03 0.10 0.27 0.03 0.04 0.14 0.01 0.29 0.04 0.12 0.15 0.59  97.80  = unknown/ u n i d e n t i f i e d f a t t y a c i d s = a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  177 APPENDIX 4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.7: F a t t y a c i d p r o f i l e s (mean and s t a n d a r d e r r o r o f t h e means) o f TOTAL LIPIDS i n t h e eggs o f 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l grower d i e t (COMM) o r on a b r o o d (BROOD) d i e t p r i o r to t r a n s f e r t o f r e s h w a t e r f o r spawning. (n=5 f i s h f o r b o t h g r o u p s ) Diet  Fatty  COMM  BROOD  Acid  mean %  sem  14: 0 14:ln9 1 ** 16: 0 16:ln9 16:ln7 5 ** 18:0 18:ln9 18:ln7 18:ln5 18:2n6 18:3n6 18:3n4 18:3n3 18:4n3 18:4nl 20:lnll 20:ln9 20:2n6 20:3n6 20:4n6 20:4n3 20:5n3 22:lnll 21:5n3 22:4n6 22:5n3 22:6n3  1.95 0.38 0.25 11.11 0.75 6.35 0.50 3.96 27.42 3.73 0.45 0.38 4.65 0.43 0.16 0.70 0.84 0.20 0.68 1.58 0.40 0.69 1.54 0.73 8.11 0.34 0.35 0.13 3.32 16.38  0.10 0.07 0.01 0.22 0.08 0.06 0.0 3 0.07 0.74 0.08 0.01 0.10 0.19 0.09 0.05 0.05 0.08 0.05 0.05 0.09 0.04 0.02 0.02 0.06 0.45 0.03 0.04 0.06 0.22 0.33  Total  98.43  ~j  * *  a.  «5L  mean % 2.12 0.64 0.25 11.72 0.39 to 7.02 to 0.49. 4.17 26.17 3.66 0.44 0.23 4.80 0.29 0.13 0.70 0.60 to 0.20 0.40 to 1.85 to 0.47 0.78 1.64 0.70 8.29 0.28 0.24 0.19 3.47 15.58  sem 0.08 0.12 0.01 0.32 0.12 0.18 0.03 0.11 0.21 0.18 0.01 0.04 0.20 0.02 0.05 0.04 0.05 0.05 0.07 0.06 0.02 0.04 0.07 0.03 0.15 0.03 0.03 0.04 0.08 0.14  97.90  ** = unknown/ u n i d e n t i f i e d f a t t y a c i d s ab = a s i g n i f i c a n t d i f f e r e n c e (a = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  178 APPENDIX 4  Fatty  Acid  P r o f i l e s o f Eggs  T a b l e A4.8: F a t t y a c i d p r o f i l e s (mean and s t a n d a r d e r r o r o f t h e means) o f POLAR LIPIDS i n t h e eggs o f 4 - y e a r o l d b r o o d s t o c k on a c o m m e r c i a l grower d i e t (COMM) o r on a b r o o d (BROOD) d i e t p r i o r t o t r a n s f e r t o f r e s h w a t e r f o r spawning. (n=5 f i s h f o r b o t h g r o u p s ) Diet Fatty Acid  COMM  BROOD  mean %  sem  mean %  sem  14: 0 14:ln9 j_ * * 16:0 16:ln9 16:ln7 4 ** 4a * * 18:0 18:ln9 18:ln7 18:ln5 18:2n6 20:lnll 20:ln9 20:ln7 20:2n6 20:3n6 20:4n6 20:3n3 20:4n3 20:5n3 22:lnll Y ** 22:4n6 22:5n3 22:6n3  1.02 2.14 0.26 15.62 0.54 2.06 1.74 0.30 8.03 13.67 3.67 0.52 2.02 0.28 1.95 0.16 0.47 0.77 2.56 0.91 0.42 10.77 0.16 0.10 0.36 3.49 24.27  0.04 0.46 0.04 0.28 0.04 0.08 0.62 0.01 0.22 0.38 0.12 0.03 0.09 0.04 0.15 0.05 0.06 0.06 0.05 0.29 0.04 0.28 0.08 0.05 0.05 0.18 0.40  1. 01 2.39 0.27 16.33 0.10 2.19 0.16 0.25 8.51 13.15 3.89 0.52 1.90 0.14 2.29 0.08 0.56 0.88 2.67 0.00 0.35 11.00 0.05 0.35 0.54 3.64 23.89  0.05 0.79 0.01 0.54 0.10 0.10 0.10 0.07 0.21 0.08 0.09 0.05 0.05 0.06 0.03 0.05 0.04 0.04 0.09 0.00 0.04 0.33 0.05 0.07 0.04 0.06 0.38  Total  98.27  a.  EK  a. a.  to to  to  to to  97.10  ** = unknown/ u n i d e n t i f i e d f a t t y a c i d s ab = a s i g n i f i c a n t d i f f e r e n c e ( a = 0.05) was f o u n d between means w i t h d i f f e r e n t s u p e r s c r i p t s w i t h i n t h e same row.  

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