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Textural and chemical changes in the muscle of chum salmon (oncorhynchus keta) during spawning migration Reid, Rebecca Anne 1991

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TEXTURAL THE  M U S C L E OF  AND  CHEMICAL  CHUM SALMON  DURING SPAWNING  CHANGES  (ONCORHYNCHUS  KETA)  MIGRATION  By REBECCA B.S.Ag.,  THESIS THE  ANNE  REID  The U n i v e r s i t y  SUBMITTED  of  Saskatchewan  IN PARTIAL FULFILLMENT  REQUIREMENTS MASTER  FOR  OF  THE  DEGREE  OF  SCIENCE  in THE  FACULTY  OF  (Department  We  accept to  THE  this  o f Food  thesis  the required  UNIVERSITY  OF  May (c)  GRADUATE  Rebecca  as  STUDIES  Science)  conforming  standard  B R I T I S H COLUMBIA 1991  Anne  Reid,  1991  OF  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 agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the head of my department  or  by his  or  her  representatives.  It is  understood  that  copying or  publication of this thesis for financial gain shall not be allowed without my written permission.  Department of  ^hrrcyi  S'CAS^  The University of British Columbia Vancouver, Canada  DE-6 (2788)  r<?^  ABSTRACT  The  spawning  deterioration a  loss  in  the  the  white-grey, texture  of  a  eating  colour  spawning  the  and  were  fish  watery.  sampled in  intermediates,  the  Grade  astaxanthin  three,  in  There  and  the  appeal  a is  red-pink  to firm  of  economic l o s s of  texture  instrumentally  the  raw  lipid  and  of  with  texture  stages  describe  during the  occurred  the this  the  the  an  and  grades  fish, were  were  increase  Hunter  fish based one,  grade  two,  progressively  the  most a  mature.  significant  i n moisture.  values  a/L,  a  and  The a/b  i n c r e a s i n g grade. canned  fish  Universal  a n a l y s i s (TPA)  judges a l s o evaluated fibrousness,  darks,  the  Grade  muscle r e v e a l e d  Instron  profile  mature  their  physical, as  into four  spawning,  decreased s i g n i f i c a n t l y with  firmness,  from  reduced  four  to  least  four,  d e c r e a s e i n p r o t e i n and  its  at  order  grade  Proximate a n a l y s i s of  semi-trained  flesh.  c h a r a c t e r i s t i c s .  were  and  more m a t u r e .  the  The  were c a t e g o r i z e d  external  silver-brights,  using  muscle  s t r u c t u r a l changes which  The  their  The  the  results  o f f - f l a v o u r develops  and  migration  matured.  of  salmon  commodity.  Chum s a l m o n  level  chum  f r e s h p r o d u c t r e s u l t s i n an  i m p o r t a n t B.C.  chemical  of  q u a l i t y of  of  distinct  becomes s o f t  c a n n e d and  on  migration  the  d r y n e s s , and  -ii  was  measured  Testing  method.  Machine  A panel  of  canned f l e s h based  on  chewiness.  The  sensory  panel  significantly spawning  one  The  fish  by  by  appearance  four  of  myofibril  filaments,  Proteinase 3.5,  activity  6.2 at  and pH  p h o s p h a t a s e , significantly lysosomal the  or  and  an  actin  7.0. but  enzymes  were  than  the  parameters, between  between  the  suggested  that  sensory  panel  an  within  smearing, myosin,  in  of  the  was  muscle  not  a  were  microscopic apparent  muscle  the  in  the  as  the  myofibrils  Changes  loss  in  loss  of  of  splitting, loss  level.  fibres  degeneration  thinning,  a  distinct  the  myosin  were  tearing or  actin  i n t e r m y o f i b r i l space.  the  dorsal  There  was  no  change  grade.  of  micrographs.  lysosomal  with  TPA  texture  showed  Focal  and  activity  a  of the  number  increase  3.5  chewy  distinguish  electron  electron  bundles,  and  to  sections  criteria,  the  and  c o r r e l a t i o n  by  clearly  mature.  the  able  measure  light  diameter  in  dry,  fish  Instron.  the  more  silver-bright  instrumental  were  u l t r a t h i n  at  evident  described  pH  the  the  Poor  sensory  micrographs  became  2  four.  and  myofibrillar  ten  characterized  measured  light  the  slope  and  photographed  firm,  Of  and  Thin  of  fibrous,  softening  being  was  more  and  instrumental the  that  fish.  cohesiveness grade  determined  a  muscle  significant at  pH  marker, These  i i i  6.2  to  or  a l s o  findings  largely contributed  muscle.  was  the  assayed increase 7.0.  at in  Acid  i n c r e a s e d  suggested degradation  that in  TABLE OF CONTENTS  ABSTRACT  .  LIST  OF T A B L E S  LIST  OF FIGURES  i  v i i i x  ACKNOWLEDGMENTS  LITERATURE  ,  1  1.1.  Introduction  1.2.  Biochemical  1.3.  1.4.  1.5.  1 changes  i n t h e muscle  o f chum  salmon  Colour  2 4  Texture  8  1.3.1.  Instrumental  1.3.2.  Sensory  Ultrastructure  analysis  8  analysis  o f t h e muscle  1.4.1.  S t r u c t u r e o f muscle  1.4.2.  Muscle  1.4.3.  Starvation  Proteinase  CHEMICAL 2.1.  x i i  REVIEW  1.2.1.  i  degeneration  18 o f chum  salmon  ..  23 23  *  29, 33  activity  36  1.5.1.  Lysosomal  system  1.5.2.  Protein turnover  1.5.3.  Starvation  36 ,  37 39  ANALYSIS  42  Introduction  42  -iv-  2.2. M a t e r i a l s a n d M e t h o d s  43  2.2.1. S a m p l i n g  43  2.2.2. G r a d i n g  43  2.2.3. P r o x i m a t e  analysis T o t a l  lipid M o i s t u r e  44 analysis  ........  analysis  44 46 P r o t e i n d e t e r m i n a t i o n  46  2.2.4. C o l o u r  47 A s t a x a n t h i n measurement  47 H u n t e r L a b measurements  47  2.3. R e s u l t s a n d D i s c u s s i o n  48  2.3.1. A s t a x a n t h i n c o n t e n t  48  2.3.2. H u n t e r L a b v a l u e s  50  2.3.3. P r o x i m a t e  54  analysis  3. TEXTURE ANALYSIS  57  3.1. I n t r o d u c t i o n  57  3.2. M a t e r i a l s a n d M e t h o d s  60  3.2.1. I n s t r u m e n t a l 3.2.2. S e n s o r y  analysis of texture  analysis of texture  3.3. R e s u l t s a n d D i s c u s s i o n  ....  61 62 65  3.3.1. I n s t r u m e n t a l  65  3.3.2. S e n s o r y  73  analysis of texture  -v-  3.3.3.  4.  PROTEINASE  Sensory  and i n s t r u m e n t a l  correlation  ACTIVITY  84  4.1.  Introduction  84  4.2.  Materials  87  and Methods  4.2.1. M e a s u r e m e n t o f p r o t e i n a s e 4.2.2. 4.3.  5.  Results  and D i s c u s s i o n  4.3.1.  Proteinase  4.3.2.  Acid  STRUCTURAL  activity  phosphatase  Introduction  5.2.  Materials  5.2.2. Results  5.3.2.  87  activity  89  90  activity  93  99 99  and Methods  100  microscopy  100  Electron microscopy  100  and D i s c u s s i o n  5.3.1. L i g h t  ..  90  CHANGES I N M U S C L E  5.1.  5.3.  activity  Measurement o f a c i d phosphatase  5.2.1. L i g h t  6.  78  101  microscopy  Electron  101  microscopy  ,  CONCLUSIONS  104  120  7 . REFERENCES  .  8. A P P E N D I X E S  125  134  vi*  LIST  OF  TABLES  Table  l . l .  Content and percentage composition of the major carotenoids i n t h e chum s a l m o n m u s c l e d u r i n g s p a w n i n g m i g r a t i o n (Ando a n d Hatano, 1987).  Table  2.1  Mean v a l u e s f o r a s t a x a n t h i n c o n t e n t t h e d o r s a l m u s c l e o f chum s a l m o n .  Table  2.2  Hunter L a b mean d o r s a l m u s c l e o f chum  Table  2.3  R e s u l t s from t h e c o r r e l a t i o n a n a l y s i s between grade, Hunter L a b v a l u e s and astaxanthin content.  ....  53  Table  2.4  Mean v a l u e s f o r percentage protein, moisture and l i p i d content of dorsal m u s c l e o f chum s a l m o n o n a w e t b a s i s .  . —  55  Table  3.1  Mean i n s t r u m e n t a l TPA p a r a m e t e r s t h e f o u r g r a d e s o f chum s a l m o n .  of  ....  67  Table  3.2  P r o b a b i l i t y values produced one-way ANOVAs on t h e s g u a r e t r a n s f o r m e d I n s t r o n TPA d a t a .  from root  ....  69  Table  3.3  Mean u n t r a n s f o r m e d results from t h e .... sensory panel's e v a l u a t i o n of the t e x t u r a l d i f f e r e n c e s between t h e f o u r g r a d e s o f chum s a l m o n .  74  Table  3.4  R e s u l t s from t h e a n a l y s i s o f v a r i a n c e of the sensory analysis to determine d i f f e r e n c e s between grades and s e q u e n t i a l r e j e c t i v e B o n f e r r o n i procedure f o r the multiple comparison of the data.  77  Table  3.5  Correlation values (r) extracted from a Pearsori c o r r e l a t i o n m a t r i x of the comparison between t h e i n s t r u m e n t a l parameters and t h e sensory attributes.  79  Table  4.1  P r o t e i n a s e a c t i v i t y a t p H 3 . 5 , 6.2 a n d 7.0 d u r i n g t h e d i f f e r e n t stages of spawning m i g r a t i o n .  91  - v i i -  values salmon.  ....  6  i n ....  48  f o r t h e ....  51  ....  Table  4.2  Acid phosphatase a c t i v i t y phosphate) i n d o r s a l muscle o f salmon d u r i n g spawning m i g r a t i o n .  Table  5.1  Percentage of silver-bright a n d .... 119 spawning f i s h a s s i g n e d t o each s c o r e u s e d t o j u d g e m i c r o g r a p h s o f chum d o r s a l muscle.  Table  8.1.  A b s o r b a n c e a t U.V. 280 o f s o l u b l e N .... 134 v e r s u s grade as a measure o f t h e p r o t e i n a s e a c t i v i t y o f chum d o r s a l m u s c l e a t pH 3.5.  Table  8.2.  A b s o r b a n c e a t U.V. 280 o f s o l u b l e N .... 135 v e r s u s grade as a measure of the p r o t e i n a s e a c t i v i t y o f chum d o r s a l m u s c l e a t pH 6.2.  Table  8.3.  A b s o r b a n c e a t U.V. 280 o f s o l u b l e N .... 136 versus grade as a measure o f t h e p r o t e i n a s e a c t i v i t y o f chum d o r s a l m u s c l e a t pH 7.0.  Table  8.4.  C o n c e n t r a t i o n o f p h o s p h a t e (mM) v e r s u s .... 137 grade as a measure of the acid p h o s p h a t a s e a c t i v i t y o f chum m u s c l e d u r i n g spawning m i g r a t i o n .  -viii-  (mM .... chum  95  LIST OF FIGURES F i g u r e 1.1  Generalized texture p r o f i l e (TPA) curve (Bourne, 1982).  a n a l y s i s ....  15  F i g u r e 1.2  Texture p r o f i l e 1982)  (Bourne, . . . .  22  F i g u r e 1.3  A p p e a r a n c e o f m u s c u l a t u r e o f a .... typical fish. White spaces r e p r e s e n t the p a r a l l e l muscle c e l l s , "dividing" l i n e s r e p r e s e n t t h e myocommata which separate the myomeres (Love, 1980).  25  F i g u r e 1.4  Major s u b c e l l u l a r components myotomal muscle ( P e t e r , 1973).  o f ....  27  F i g u r e 2.1  Illustration chum salmon.  of ....  45  F i g u r e 3.1  Example o f t y p i c a l t e x t u r e s c o r e .... s h e e t used t o a n a l y z e t e x t u r e o f t h e f o u r g r a d e s o f t h e c a n n e d chum samples.  64  F i g u r e 3.2  Example o f a t y p i c a l I n s t r o n t e x t u r e . . . . p r o f i l e a n a l y s i s c u r v e f o r a chum sample.  66  F i g u r e 3.3  Standardized sensory scores f o r the .... f o u r g r a d e s o f chum s a l m o n f o r t h e attributes firmness, chewiness, dryness arid f i b r o u s n e s s .  76  F i g u r e 4.1  Flow d i a g r a m o f t h e method u s e d t o . . . . measure t h e p r o t e i n a s e a c t i v i t y i n the d o r s a l muscle o f f o u r g r a d e s o f chum muscle.  88  F i g u r e 4.2  Standard curve o f the absorbance a t .... 4p5 nm v e r s u s t h e c o n c e n t r a t i o n o f E-nitrophenol;  94  F i g u r e 5.1  L i g h t m i c r o g r a p h o f a c r o s s s e c t i o n . . . . 102 of s i l v e r - b r i g h t chum salmon s t a i n e d with p h o s p h o t u n g s t i c acid h a e m a t o x y l i n . ( M a g n i f i c a t i o n 1500).  score  sheet  of the four  -ix-  grades  Figure  5.2  L i g h t m i c r o g r a p h o f a c r o s s s e c t i o n .... 103 o f s i l v e r - b r i g h t chum s a l m o n s t a i n e d w i t h p h o s p h o t u n g s t i c a c i d haematoxylin. ( M a g n i f i c a t i o n 600).  Figure  5.3  L i g h t m i c r o g r a p h o f a l o n g i t u d i n a l .... 105 section of a silver-bright chum salmon s t a i n e d w i t h p h o s p h o t u n g s t i c acid haematoxylin. (Magnification 1500).  Figure  5.4  L i g h t m i c r o g r a p h o f a c r o s s s e c t i o n .... 106 o f a s p a w n i n g chum s a l m o n stained w i t h p h o s p h o t u n g s t i c a c i d h a e m a t o x y l i n . ( M a g n i f i c a t i o n 1500).  Figure  5.5  L i g h t m i c r o g r a p h o f a c r o s s s e c t i o n .... 107 o f a s p a w n i n g chum s a l m o n stained w i t h p h o s p h o t u n g s t i c a c i d haematoxylin. ( M a g n i f i c a t i o n 600).  Figure  5.6  L i g h t m i c r o g r a p h o f a l o n g i t u d i n a l .... 108 s e c t i o n o f a s p a w n i n g chum s a l m o n stained with phosphotungstic acid haematoxylin. ( M a g n i f i c a t i o n 1500).  Figure  5.7  Electron micrograph o f t h e d o r s a l .... 109 muscle of a s i l v e r - b r i g h t chum salmon. ( M a g n i f i c a t i o n 27000).  Figure  5.8  Electron micrograph o f t h e d o r s a l .... 110 muscle of a s i l v e r - b r i g h t chum salmon. ( M a g n i f i c a t i o n 27000).  Figure  5.9  Electron micrograph o f t h e d o r s a l .... muscle of a s i l v e r - b r i g h t chum salmon. ( M a g n i f i c a t i o n 27000).  Figure  5.10  E l e c t r o n m i c r o g r a p h o f t h e d o r s a l .... 112 muscle of a s i l v e r - b r i g h t chum salmon. ( M a g n i f i c a t i o n 27000).  -x-  I l l  F i g u r e 5.11  Electron micrograph of the dorsal m u s c l e o f s p a w n i n g chum salmon. Arrow i n d i c a t e s area o f "smearing". ( M a g n i f i c a t i o n 26000).  F i g u r e 5.12  Electron micrograph of the dorsal m u s c l e o f s p a w n i n g chum salmon. Arrows i n d i c a t e areas o f " s p l i t t i n g " of t h e m y o f i b r i l s . (Magnification 26000).  F i g u r e 5.13  Electron micrograph of the dorsal m u s c l e o f a s p a w n i n g chum s a l m o n . Notice areas with " i n t e r m y o f i b r i l l a r " s p a c e s . ( M a g n i f i c a t i o n 26000).  F i g u r e 5.14  Electron micrograph of the dorsal m u s c l e o f s p a w n i n g chum salmon. Arrows p o i n t t o areas which have myofibril "thinning". (Magnification 26000).  -xi-  ACKNOWLEDGMENTS  No  one c o u l d  ever claim  that  individual  effort.  and  h e l p e d me c o m p l e t e t h i s  advice  sharing  project,  encouragement  I would  t h a n k s t o w a r d s my s u p e r v i s o r ,  f o r h i s patience,  encouragement.  was e n t i r e l y a n  Of t h e many p e o p l e who's  e x t e n d my most s i n c e r e Durance,  a thesis  I would a l s o  h i s invaluable  knowledge  and  D.  Kitts  l i k e t o t h a n k D r . D. W a l k e r f o r  knowledge  meetings ideas  into  a n d D r . W. their  a n d comments.  busy  D r . T.  constant  of electron  m i c r o s c o p y and  f o r t h e S a t u r d a y s h e s p e n t h e l p i n g me w i t h my work. Dr.  like to  Powrie  f o r fitting  committee  s c h e d u l e s and f o r t h e i r  I would  also  like  I thank  t o t h a n k my  helpful father  f o r h i s h e l p i n t h e l a b a n d f o r h i s c o n f i d e n c e i n my work. It author  i sfitting Douglas  t o end t h i s  Adams,  s e c t i o n w i t h a quote from t h e  "So L o n g ,  Fish".  -xii-  and Thanks  fora l l  the  1. LITERATURE  1.1.  Introduction  Chum  salmon  cial  commodity  fishery. landed  i n B.C.,  Chum  o f 27 m i l l i o n  are hatched  after  emerging  to  t h e s e a , where t h e y  on  plankton  may  90 t o 9 9 % o f t h e s t o c k  may  begin  depending often  i n an advanced  their  natal  (Neave,  1966).  they  stop  tein  i n their A  is  a  loses  result  loss  white  Bilinski  colour.  soft  to five  Spawning third  years  late  quality  Chum a r e  as they  enter  who  These  -1-  and pro-  i n t h e salmon The  ultimately from  changes  studied  water,  spawned.  changes  The t e x t u r e changes  enter  the sea  fresh  of the fish.  and  January,  n o t f a r from  have  begins  year and  as  1982).  maturity  they  feeding  migration  or fourth  as  spawn  until  running  downstream  t h e salmon  colour,  (1984)  fast  s u r v i v e on s t o r e d l i p i d  and watery.  et a l .  total  t h e f r yd r i f t  physiological  i n the eating  f o r a  a few days  of sexual  before  of  and S t a r r ,  and o f t e n  muscle  i t sred-pink  to  (Beacham  of these  salmon  Within  or extend  f e e d i n g , and must  white-grey able  Even  beds  three  i n their  stage  streams,  beds,  spend  as J u l y ,  on t h e s t o c k  t h e #3  caught  the winter.  crustacean.  for  as  commer-  dollars.  t h e spawning  as e a r l y  were  i n the gravel  of  and small  i s an important  i t ranks  tonnes  incubating over  from  keta)  where  I n 1987, 11,000  value  streams  fOncorhynchus  REVIEW  firm were  the effect  muscle  becomes and  desir-  noted of  a  by  sexual  maturation These an  on  the  workers  noticed  increase  i n  discolouration tion  of the The  uration fish.  of  quality could  weaker  this  Ando  and  f a t and assess the  (feeding  non-protein,  sarcoplasmic substances  during  used  spawning  as  a  fisheries the  a  deteriora-  the  begin  mat-  ocean-run  stocks  cannot  their  be  spawning  i n the over  harvesting  management  techniques  fishing  measured of  the  At  muscle  and  low  fleet  to  The  reversed and  lipid  stages energy  of  i n the  muscle  state  of  of  first high  target  As so  the that  the  migration, f o r gonad  colour, chum  in  salmon  stage  tested  i n  total  and  p o s i t i v e ,  spawning the  pro-  total  ninhydrin was  salmon  the  ninhydrin  content  source  -2-  the  the  i n  o f chum  changes  was  compounds.  decreased,  early  as  free o i l ,  only  i n the muscle  content  relationship  primary  they  period.  protein,  the  such  i n  and  different  physiological  increased.  product  1982).  moisture  protein  salmon.  i n t h e chum d u r i n g  result  allow  nitrogenous  this  flesh  occur  may  (1986a)  the  coho  decrease  the  of  changes  migration),  sarcoplasmic  gressed,  and  Hatano  canned  harvesting  fish  (Holmes,  Biochemical  throughout  by  Terminal  problem  stocks  to  of  which  fisheries  1.2.  order  a  of  odour.  because  stocks.  protein,  duced  softening  avoided  stock  flesh  i n the  distinguished until  mixed  specific  was  be  the  l i q u i d ,  changes  However,  reduce  free  and  of  changes  f l a v o u r and  adequately run,  quality  and  positive  markedly  re-  indicating  i t  development.  Idler  and  content and  no  Bitners  of  the  change  (I960)  ovaries, i n ovary  run.  Analysis of  tein,  a  slow,  moisture  glycogen  as  in  an  200%  increase i n the  increase by  the  revealed  increase  state  the  of  the  female  early  migratory et  (1960)  indicator  however  f o r both  in  a  in  ovary of  33%  increase  f a t , and  fat  moisture,  end  as  a  the  the  migratory  little  sockeye  significantly the  and  level  increased  found  of  the  in  pro-  change  that  the  term  of  in  lower  the  and  Glycogen  than  decreased  in  the  during  the  later  carbohydrate  stage.  stores  migration,  emergency  liver  condition  salmon.  in  throughout  short  level  physiological  spawning  maintained  source  monitored  sexes  phase,  were  of  were  a l . (1980)  muscle  tioned  milt  Idler  male,  the  11%  protein,  the  steady  and  nutritional  Mommsen  an  a  content.  Chang  levels  measured  and  energy  in  funcin  the  muscle. Duncan  and  actomyosin,  rily  in  to  nitrogenous  the  the  Ando  (1958)  myoalbumin  non-protein crease  Tarr  total  and  myogens,  material  nitrogen  decrease and  studied three  i n actomyosin  Hatano  (1986b)  by  N - m e t h y l h i s t i d i n e by  This  protein  amino  degradation  acid  degradation  and  i s useful  because  as  i t i s  -3-  was  collagen,  ion an  muscle.  stroma  during  fractions,  attributed  followed  protein  phy.  and  sockeye  content  myofibrillar measuring  in  protein  and A  de-  prima-  fraction. the  degree  spawning  exchange index  contained  of  of  migration  chromatogramyofibrillar  only  in  actin  and  myosin,  excreted  i s not  re-utilized  quantitatively  workers  were  protein  able  to  into  urine.  clearly  degradation  for protein synthesis, In  show t h e  during  this  degree  spawning  and  manner, of  is  these  myofibrillar  migration  of  chum  salmon. 1.2.1. Carotenoids such and  a  group  Pacific  salmon,  red  colours  found  Astaxanthin  main  pigment  salmon  are  from  the  animal  The salmon the  HPLC  found  diet.  Thus  a  levels, ries.  the  an  important salmon  the  the  the  closely  fish  associated  (Ando  et  to  throughout  With  an  the  increase  4-keto-zeaxanthin  the  spawning  there  sexual  1985).  follow  carotenoid  migration and  a l .  was  and the  an  skin  zeaxanthin  -4-  be  in  the  metabolism  colour  a l .  change  in  colour  of  of  chum  (1989) of  used chum  Astaxanthin  immature in  chum  serum  levels  considerable were  novo,  obtained  et  carotenoid a  de  physiological state  increase  i n the  Because  Ando  in  the  a l . , 1989).  migration.  found  maturity,  and  with  et  and  ova-  i s  pigmentation  (Ando  muscle  and  1982).  must  ingestion  differences  orange, and  carotenoid  of  animals  yellow,  carotenoid,  this  in  muscle,  carotenoids  sources  in  the  (Simpson,  degree  of  to  which,  integument,  related  reflection  primary  During  in  d i e t a r y pigment  are  pigments  synthesize  other  techniques  salmon  to  of  contribute  i n chum  and  i s  specific  i s  unable  astaxanthin  the  are  as  ries.  of  Colour  detected  was  salmon.  carotenoid i n the  ova-  proportion i n the  of  muscle  of  the  was  spent  via  a  chum.  The  reductive  4-keto-zeaxanthin loss  in  and  to  and the  the  presence  number  of  of  of  a  the  of  hue  the of  the  the  carotenoids  in  The s h i f t  acidity,  causes  astaxanthin  is  an  quantification  posure  to  light,  There  is  consistent colour when  a  a  are  cis  a  need  for  or  it  and  chemical  and p h y s i c a l  spectral  sensitivity  properties  oped  quantitate  the  was  the  from  of  its  result  bonds.  of  As  the  the  wavelengths, In  general,  configura-  heat,  colour.  is  important  excess  of  maximum  of  it  heat,  techniques  1986).  composition  of  characteristics  colour by  a  trans  a result  light, Because during  acid,  the  -5-  of  the  to  ex-  is  the of  the  response Commission  on  perceived  related  light  eye.  provide  information  The c o l o u r  illuminated object  proposed  is  longer  quantitative  the  eye  as  instrumental  factors,  human  this  lipoxygenases.  eye  spectral  to  zeaxanthin  stages  redder.  in  pigment,  the  to  an  the  becomes  and Storebakken,  views  in  four  double  lightening  protect  oxygen,  depicts  increase,  at  found  gradual  to  1.1  carotenoids  bonds  isomer  unstable  change  astaxanthin  increase  conjugated  double  qualitative  (Skrede  of  carotenoid  to  Table  this  1987).  different  food  for  from  chum d u r i n g  compound o c c u r s  the  any  the  system  and  tion.  in  conjugated  absorbance  pathway  relative  (Ando and H a t a n o , colour  proposed  zeaxanthin.  4-keto-zeaxanthin  The  or  metabolic  astaxanthin,  migration  mechanism  to  three  source,  object, A system  mechanism  and  the the  develof  International  the de  Table  1.1.  Grade  Sex  Astaxanthin  Zeaxanthin  4-ketozeaxanthin  M F  14.5 11.4  (89) (91)  0.19 0.10  (1.2) (0.8)  0.34 0.20  (2.1) (1.2)  1  2  Content and p e r c e n t a g e composition of the major c a r o t e n o i d s i n t h e chum s a l m o n m u s c l e during s p a w n i n g m i g r a t i o n (Ando and H a t a n o , 1 9 8 7 ) .  1  2  M F  10.7 9.4  (86) (84)  0.32 0.31  (2.6) (2.8)  0.45 0.48  (3.7) (4.3)  3  M F  0.4 0.4  (73) (78)  0.07 0.04  (11.8) (8.4)  0.03 0.02  (5.0) (4.3)  4  M F  0.7 1.0  (64) (70)  0.26 0.29  (23.7) (20.0)  0.05 0.05  (4.9) (3.4)  Brackets indicate percentage of t o t a l carotenoid. V a l u e s a r e m e a s u r e d i n mg/100 g w h o l e m u s c l e .  -6-  l'Eclairage scales mate  were  not able  of colour  scales  were  colour  posed  as perceived  opponent  eye  between nerve,  switching  which  stage,  a  t h i r d  yellow  the CIE  uniform  esti-  eye, other  on t h e opponent  colour  colour theory  describe  a  of  L a b  makes u s e o f a  flected  response  with  from  i s used  was  colour  related  b y G.E.  The t h e o r y  stated  that i n  i n the retina  then  was  and t h e  to the brain. were  In the  compared  dimension, compared  dimension,  with  l i g h t n e s s ,  colour.  as "b". ( L ) , a  reflectance  a n IBM p e r s o n a l  used  successfully  model  of this  spectrophotometer  computer.  i s analyzed  Visible  values  to ma-  which i s light re-  and t h e r e s u l t a n t  the spectral  have  information.  The modern  reflectance  as  blue to  of colour classification  has been  with  denoted  denoted  t o the percent  quantitative  system  to calculate  pro-  refined  colour  systems  t h e sample  was  later  signals  considered  to provide  vision  intermediate signal-switching  red-green  t o blue  and measure  interfaced  an  colour  Several  Hunter  of colour  receptors  f u n c t i o n which  developed The  took  red-green  an o b j e c t .  trum  t h e human  1980).  was  dimension  non-linear  chine  by  because  reasonably  the r e d colour responses  This  been  a  theory  the light  "a".  of  However  i n 1 8 7 9 , a n d was  there  t o generate  generate  colour  ( d e Man,  green  A  based  Hering  i n 1930  human  optic  to provide  developed  by Ewald  Mueller  stage  1980).  vision.  The  the  ( C I E ) ( d e Man,  spec-  L, a, and b.  -70  1.3.  Texture 1.3.1. Instrumental a n a l y s i s  The  t e x t u r e of a food p r o d u c t  the most d i f f i c u l t rately. caused  i s commonly  organoleptic property  Szczesniak,  (1963) s u g g e s t e d  considered  to describe this  accu-  problem  was  f i r s t by the l a c k of an adequate b r i d g e between theo-  retical  rheology  and  practical  a p p l i c a t i o n s , and  second  cause most of the r e p o r t e d r e s e a r c h had only c o n s i d e r e d cific was  t e x t u r a l c h a r a c t e r i s t i c s of i n d i v i d u a l  bespe-  A need  foods.  thus i d e n t i f i e d t o develop a r a t i o n a l system, and nomen-  clature  t o d e s c r i b e and  translate textural properties into  p r e c i s e l y d e f i n e d , measurable p r o p e r t i e s (Friedman  et a l . ,  1963). Texture  and  c o n s i s t e n c y were d e f i n e d by  two  elements,  f i r s t the p h y s i c a l s t r u c t u r e of the m a t e r i a l ( i . e . i t s geome t r y ) , and (i.e.  second the way  i t s mechanical  and  the m a t e r i a l f e l t  i n the  surface properties).  (1963) grouped t e x t u r e i n t o t h r e e main c l a s s e s , characteristics to s t r e s s , the  - those manifested  geometric  mouth f e e l  factors.  acceptance  of  ters,  hardness,  elasticity,  and  five  and  Szczesniak, mechanical  by the r e a c t i o n of  characteristics  c o n s t i t u e n t s of f o o d ,  mouth  other  - the  food  arrangement  characteristics  - the  T h i s a n a l y s i s of t e x t u r e l e a d t o primary  independent mechanical  cohesiveness,  adhesiveness,  t h r e e secondary  t l e n e s s , chewiness, and  -8-  viscosity,  Hardness was  the  parame-  dependent parameters,  gumminess.  of  and brit-  d e f i n e d as  the  force necessary  ness  was  body  of  the  force, rate  the  siveness forces  after  quired  ticity.  size tion.  was  food the  force  food  was  product  geometric  and  shape Other  a  of  to  energy a  particles,  to  the  to  an  with  to  which  of  other  Brittleness, the  material  parameters  was  the  state  energy  of re-  ready  for  swal-  cohesiveness,  and  elas-  for  disintegrate a swallowing.  described their  described  perception  adhe-  attractive  surface  required to  and  and  the  primary  a  undeformed  removed,  the  s t a t e ready  characteristics  related  the  i n contact.  characteristics  the  was  and  hardness,  the to  food  the  springiness)  overcome  the  up  unit  Chewiness  solid  made  per  was  to  cohesive-  flow  returned  food  to  which  rate of  to  came  related  related  The  were  of  cohesiveness.  Gumminess  semisolid  was  masticate  and  the  was  the  force  necessary  deformation,  bonds  was  material  surface  which  and  and  to  lowing,  and  work  given  changed  deforming  fracturability,  hardness  viscosity  the  the  with  a  internal  (later  the  between  fractured,  of  deformed  was  materials or  product,  which  condition  attain  strength  elasticity  at  to  of  the  relative  relative  mouth  orienta-  feel  moisture  and  quality fat  in  design  of  food. Application an  instrumental  tions  into  Friedman  an et  of  these  unit  capable  unbiased, a l .,  parameters  ( 1963  of  translating  recorded )  required  physical  recognized  -9-  that  the  these  defini-  measurement. in  order  to  characterize  texture,  a  needed.  objective  measurement  into  The  three  tests,  general  and  and  time  such and  as  from  which  ultimate  These  failure  have  p a r a l l e l uniaxial  forces parallel  uniaxial  tangential  ting.  Compression,  tested  i n two  which  means  of  the  compression tion,  and  The  method  ple  of  action of  uniform  before (E)  The  sample  i s  the  texture  (Bourne,  i s defined  The  Young's  the  modulus  one  mechanical or  the  causes  in  of  other  Young's  1982).  -10-  and  bulk  food.  small  shear, or  cut-  can  dimensions  test  i s  be  two  by  uniaxial  i n one  direc-  dimensions. the  The  strain of  princi-  theoretical  t o Hookean  modulus  the  compression,  utilizes  applied a  uniaxial  texture,  three  second  crush  at  of  forces.  separation  is a  analysis  was  more  and  compression,  i s compressed  area by  parameters  flattening,  test  to  compression  cross-sectional  rupture,  causes  sample  profile  the  deformation,  application  compressed  i n  empirical  calculate  extension,  which  compression  uniaxial  the  first  unrestrained of  to  imposing  pressure.  whereby  force,  important measure  ways.  uniaxial of  force  an  hydraulic  by  which  divided  1978).  tension,  force  was  was  c a t e g o r y measured  instruments cause  cause  test  tests,  Poisson's ratio,  foods  to  texture  of  possible  (Bourne,  of  included  first  properties  strength,  measuring  of  objective  fundamental  The  i t was  s h e a r / b u l k modulus  structural  in  tests.  rheological  Texture  a  categories,  imitative  fundamental  comprehensive  solids  ( i . e . 1%) elasticity  E  =  Compressive Compressive  stress strain  =  F/A delta L/L  Where: Stress = force per unit area Strain = deformation per unit dimension F = applied force A = cross sectional area L = unstressed height d e l t a L = change i n h e i g h t due t o F  Thus measure ever,  "E" i s t h e slope  of stiffness  t h e pure  because amount  (Bourne,  definition  first, of  of the stress/strain  and  i s usually  second,  viscoelastic,  not elastic.  weight  on a  placed  stantaneous continuous  elastic  with  a  recovery  deformation,  height.  nition  causes  immediate  deformation"),  r e s p e c t t o time,  inal  because  most  Viscoelasticity  When t h e w e i g h t  elastic  permanent  subjected  In order  called  by  never  t o maintain most  by a  a  recovery Thus  i n a  of the defii n s t e a d by  e t a l . , 1980).  r e s i s t a n c e t o deformation.  sions  a  returns to i t s orig-  overall  on t e s t  when  "retarded  future  of deformability i s representative of a  dependent  are  the instantaneous  modulus  are  large  prolonged,  called  are defined  o f d e f o r m a b i l i t y " (Johnson  to a  occurs  the "purity"  foods  This i s  foods  "retarded recovery".  t h e sample  o f Young's Modulus,  "modulus  i s followed  how-  compression ( " i n -  followed  i s removed,  and a  foods  applies.  decelerating rate of deformation  deformation". partial  food  curve,  F o r most  of "E" rarely  t h e sample  strain,  1982).  • -  The  material's  Actual  measured  values  conditions, especially  specific  dimen-  o f t h e m a t e r i a l , and deformation  -11-  rates.  I t should  be  noted  however  caused then  a  that  product  the  modulus  (Bourne,  to of  correlate  not  and  strain  all  tests  eters,  to  sensed  be  would  only  of  no  compression or  break,  longer  i n the  Imitative conditions struments  to that  Farinograph, Universal  used  apply  these  small  they  were food  into  this  Amylograph, Machine  and  funda-  basis  from  and  tests  et  have do  1972).  been not  found  include  description  include  the not  have  of  and  tended  mechanical provided  of  penetrom-  tenderometers,  to  proper-  a  complete  texture  were  a l . , 1963). to  subjected category  imitate i n the  include  Alveograph,  (Szczesniak,  -12-  which  Bourne,  they  parameters  developed was  of  complete  have  have  function  that  but  of  not  stress  instruments  phase  (Friedman  tests  Testing  parameters  f o r these  does  often  of  a  (Brinton  mechanical  mouth  which  provide  tended  are  independent  primary  for a  have  and  food  consistometers,  a  the  are  quality,  Because  fall  that  necessary  on  because  to  measure  modulus  evaluation,  developed  i n chewing,  how  Young's  the  been  textural  devices.  involved  picture  may  compressors,  concentrate  Thus  often  Instruments  shearing  ties  uniaxial fracture,  also  sensory  properties  parameters  texture.  as  themselves  tests  related  the  such  with  in  has  Empirical be  tests  conditions.  tests  empirical  to  of  spread,  deformability  rheological  mental  degree  rupture,  poorly  meaningful  simple  high  1982).  Fundamental to  i f a  and  1963).  the  actual  mouth.  the  In-  Brabender  the One  Instron of  the  first  instruments  designed  denture  tenderometer  ratory  of  (Friedman a  basic  the et  sisted  of  and  four  response a  product profile"  to  recorder,  to  measure a  of  (Friedman  measure  the  two  zontal  cross-head  within  the  sensing and  a  tor  switch  to  record  of of  chart and  a  of  full  0.2  to  and  of  a  scale  sample's  was  as  the  con-  texturometer  con-  with  a  sensing  motor,  plate,  power  a  supply,  c i r c u i t .  which  thus  The  represented  called  to  a  an the  "texture  kg  with  1972).  cm  per  which By  a  high  use  of  machine  which  drives  deflection (2N  of to  force-distance  -13-  a  of  the  pen ,  and  a  a  load  relationship.  force cell  selec-  possible  over to  d i -  hori-  and  i t is  of  speeds  sensitivity  cells,  5kN)  is  selected  minute,  a  Machine  degree  This  consists  d i f f e r e n t load  force  Testing  d i r e c t i o n at  50  system  Universal  mechanism  vertical  recorder.  500  used  c h a r a c t e r i s t i c s of  foods  Bourne,  0.2  several  a  a  in  for  strain-gauge  Instron  parts,  recording  strip  obtain  range  range  and  the  and  basic  (1963)  articulator  curve  Labo-  Technology later  Their  textural  deformation  vided  of  the  a l . , 1963).  used  (Brinton  Technology  variable-voltage  the  test",  et  precision into  a l . ,  was  was  strain-gauge a  the  under  first  an  force-distance  picture  Bourne  et  units, and  Food  instrument  "texturometer".  drive,  sample  the  mastication  I n s t i t u t e  Friedman  basic  produced  "integral  by  This  pen  method  system  by  their  variable-pulley fast  developed  a l . , 1963).  of  simulate  Massachusetts  prototype  struction  to  a  force  produce Since  a  i t s  introduction, evaluation  evaluation  profile  used  this  parameters  tained  from  Instron  analysis  i s a  t o compress  action  a  which  correlated  1982).  number  This  technique  tidimensional generalized Bourne sample  was  texture  placed  the  sample.  second  The  fracturability.  peak,  compression.  hardness  1,  The I n s t r o n  ob-  to the  approached  abruptly  reversed,  speed.  of a force-time  parameters  could  curve  This from  be  measured  evaluations  (Bourne,  o f t h e need Figure  1.1  f o r mul-  depicts  a  curve. described  between  force  of  speed,  a t t h e same  texture.  profile  descended  then  sensory  of  curves  of the  a reflection  e t a l . (1978)  cross-head  as  was  texture  to imitate the action  stroke  with  analysis  The  known  well  the  foods.  sample  stroke,  textural  of  attached  pieces  a t a constant  an upward  of  were  f o rthe  a  resulted i n the production  which  A  driven  of the compression with  range  to evaluate  bite-sized  The c r o s s - h e a d ,  continued  o f a wide  plates  jaw.  and  technique  Flat  i n a r e c i p r o c a t i n g motion  end  food.  force-deformation  twice  the  use i n the  from  the Instron.  and used  useful  technique  derived  widespread  of solid  of t e x t u r a l parameters  (1982)  profile  has gained  of textural properties  Texture  Bourne  the Instron  two  flat  at constant  at the f i r s t As was  Figure  1.1  as  follows.  plates  on t h e  Instron.  speed  and  significant  the compression formed  cross-head  -14-  peak  continued,  at the point then  compressed  reversed  was a  o f maximum  FIRST BITE  ADHESIVE  FORCE  TIME-*  1  Figure  SECOND BITE  1.1.  Generalized Texture curve (Bourne, 1982).  -15-  Profile  Analysis  (TPA)  direction,  and t h e f o r c e  area  t h e curve  under  and  represented  during the  turned ever,  a  The line  pression phase done  the  maximum  the  right  machine food  during  by t h e machine  mum  than  ture  that  profile.  area  1,  line  defined  the force  was  during  2 was  "the distance  t h e work  could  n o t be d i r e c t l y  Cohesiveness  gumminess  was  was  t h e sample  measured  the product  -16-  done  rise  from  The recov-  o f maxiseveral the tex-  by d i v i d i n g of  on  i n force  also defined  derived  to the  compression.  and i t s p o i n t  e t a l . (1978)  area t o  returned  t h e end o f t h e f i r s t  Bourne  the zero  a s t h e maximum  between  bite,  as t h e work  the positive  t h e second  zero  t o t h e com-  under  as  i n t h e second  the  at the point of  t h e work  Area  possibly  the decompression  defined  Again,  How-  substantial  under  defined  zero  compression".  parameters  by  was  was  2 was  decompression.  i n height  greater  force  was  with  a n d was t h e a r e a  compression.  of the dotted  area  during  of  t h e work r e -  of the food  3 was  food  compression.  of the curve  upward  apart,  and crush to the right  associated  Area  Hardness  second  springiness ered  bite.  The adhesive  force.  during  1 was t h e compression,  theoretically  the adhesion  i t moved  the food  line.  negative  the  as  of the first to pull  force  portion  represented  area  of the small  a n d was  negative  plate  1) was  part  of  t o compress  t o t h e machine  significant  Area  point  The p o s i t i v e  by pen response  error.  done  ( i nArea  by t h e food  caused  force  bite.  line  t o zero.  u p t o t h e maximum  t h e work  the f i r s t  dotted  returned  area  hardness  2  and  cohesiveness,  and  chewiness  was  the  product  of  gumminess  and  springiness. To was  obtain  time  information  consuming,  et  p u t e r  a t t a c h m e n t s  facing  a l . , 1982).  the  As  a  a  signals  and  force-deformation  allowed  the  a n a l y s i s of  number  of  Bourne type  et  rious  not  variation  a  cell  speed  and  texture  with  curves  areas,  and  computer was  to  Sherman  the  apparent  (1973)  Use a the  that  et  the  claimed  the  cautioned  -17-  et to  Instron extract A  to  computer  that  of a  samlarge  curves in  se-  a l . , (1982) record  the  cross-head data  solution  select  and  automatic  from  to  variable texture  operator  simplicity  computer  resulted  able  tests.  output  profile  Abbott  automatically  the  the  was  a l . , (1982)  curves  the  cell  original  was  inter-  system  a n a l y s i s of  for texture  control  the  of  By  l a r g e number  Abbott  a n a l y s i s of  distances which  Despite  and  compression  allow  of  load  interpretation.  and  r e a d o u t s  recording  i n these  they  com-  dual  from  time,  common  signal,  type  data  problem,  a  appropriate  output  this  accuracy  developed.  curves.  cautioned  which  direction,  and  However,  program  profile  problems  and  were  errors in their  load  file  short  a l . , (1978)  the  provided  the  variables.  systems  because  measuring  relatively  to  been  manually  limited  microcomputer,  capable  in a  of  graphs  a u t o m a t i c have  formed,  ples  of  with  and  solution  w i t h  parameters  Instron  force-time  d i f f i c u l t  (Abbott  force-deformation  from  the  the pro-  peaks,  then c a l c u l a t e d .  of  the  Instron,  instrumental  Shama  evaluation  of  textural  sponses  required  selected load  properties that  carefully.  influenced  properties normally (the  the instrumental  comparison  was  properties, necessary  data.  made  between  different  i n order  Thus  i t was  instrumental  of  as t h e r a t e  therefore  to simulate  of the  textural  of  loading com-  influenced  the  that  different  test  i t was  t o achieve  suggested  with  when  a  textural  conditions  relevant  be  applied  d e p e n d i n g o n how  the force  foods  sensory r e -  conditions  evaluation  increased, and  test  and r a t e  In the Instron,  increased  force-deformation  to predict  f o r each food  speed)  also  method  The magnitude  masticated.  pression  a  the instrumental  differently  cross-head  as  may  masticatory  be  pat-  terns . 1.3.2. Organoleptic the  impact  food.  of  a  which  tests  of a process  Aguilera  sensory  Sensory  consumer  "typical"  consumer  hedonic  liked/disliked intensity ples.  have  panels,  differences  used  described  three  which measured  to the product; i . e . how  and a n a l y t i c a l  to  study  types  the response  acceptance  much  panels  t h e sample  panels  of  which  was  measured  i n t h e t e x t u r a l a t t r i b u t e s o f t h e sam-  of the analytical  which  been  on q u a l i t y a t t r i b u t e s o f a  (1990)  questions,  a n d why;  The o b j e c t  frequently  operation  and Stanley  panels;  asked  analysis  merical  data  could  problem  with  the f i r s t  "quality  was  i n t h e mind  be two  panels  analyzed techniques  was  statistically. was  of the observer",  -18-  to obtain  that and  a  nuThe  commonly measured  response and  was  was  food  result  subject  processor The  sory  a  ture. poor  t o many (Lund,  comprehensive  food  t e x t u r e was t h u s  needed  t o pro-  t h e common  between  developed  method,  analyzed  that  texture  by t r a i n i n g  term  t h e machines  were  correlations  tests.  which with  of a  an e x p e r t  texture profile" The  and sensory  as t h e sensory  food taste  profiling"  geometric,  f a t and moisture  through  complete  comprehensive judges,  and order  sensory  a standard  Szczesniak  panel  was  to  pro-  objectively  to create a  thus  used  analysis  "sen-  to describe  of a of  characteristics,  scale,  eta l . ,  demonstrated  product  was  mastication.  rating  not evaluating  of appearance  method  evalua-  product.  organoleptic multidimensional o f t h e degree  of  instrumental evaluations.  o f any food  "texture  was  tex-  problem  instrumental  a technique  this  sample  mechanical,  from  the  Requirements  included  a  panel  and proper  panel  i n  first  f o r this  of  trained  procedures  e t a l . , 1963).  Different standard tural  sen-  e t a l . , (1963) n o t e d  With  (Brant  and a n a l y t i c a l  Szczesniak  good  bite  of a  f o r classifying  very  terms  the control  w e l l - d e f i n e d system  duce  the  outside  conditioning,  rational,  same p r o p e r t y  sory  of a  and suggested  (1963)  variables  previous  1982).  to evaluate  correlation  tions  o f t h e judge's  development  method  duce  the  a  rating  textural scales  intensities.  characteristics composed  These  foods  -19-  were  of foods provided  with  c a t e g o r i z e d by different  the basis  tex-  f o rthe  evaluation  of  cific  foods  that  proper  Rating force  the at a  each  required  to  a  -  a  sample  rate  state  secondary  time one  adhesiveness  over  tongue.  the  evaluation product A  of  i n terms  Use  the  (1963)  cal,  geometric  moisture  used  the  as  garding  Texture the  was  order  these  a  those  to  gumminess  with  -  a  to  liquid  rating  parameter  i t to  secondary  in  mas-  remove  the  eating;  and  from  scales  -  sample  reduce  d u r i n g normal a  a  throughout  required  method  given  was  food  a  spoon  enabled an  classes  properties  a  given  considered to  the  "unknown"  i n which  of  to  Brandt  et  t e x t u r e ; mechani-  of  a  to  fat  recording the  by  order  definite  characteristics  -20-  developed  relating  food  have  then  product.  s y s t e m a t i c means of  hard-  products.  t h r e e main  a  teeth;  force  to masticate  draw  textural  of  characteristics  ance.  to  or  persisted  force  molar  the  s h a t t e r e d ; chewiness  i n order  mouth  sensory  and  content  that  the  known  texture  al.,  or  -  encompassed  ease  so  available.  hardness  with  the  second  the  of  given of  as  Spe-  selected  always  which  required  required  comprehensive  evaluate  tural  to  force  a  -  texture.  were  substance  cracked,  density  adhered  the  were  f o r swallowing;  tication,  -  scales  parameter  per  of  included:  defined  chew  d e f i n e d as  viscosity  a  i n seconds  parameter  that  the  and  penetrate  satisfactory  material  on  developed  crumbled,  of  parameters  standards  cohesiveness,  length of the  point  reference were  and  which  for  mechanical  scales  brittleness ness  the  were  of  and tex-  appear-  pattern reperceived.  First The  bite,  first  m a s t i c a t o r y , and  bite,  or  residual  initial  phase,  characteristics  of  other  characteristics  geometric  second  bite,  or  characteristics other The  duced  bite,  i n  the  throughout  selection  judges  food  tural  of  sheet  that  tained  mechanical  during  encompassed  geometric  based  foods  panel  and  chewing.  changes  in-  characteristics  be  i s shown trained an  interest was  accurately  were were  The  and  until  A  unknown  individually  the  The  for a  scale.  sample  typical  the  geometric  panel  and  then  texchar-  devel-  using the  pre-  texture profile  1.2.  objective of  familiarize  c l a s s i f i c a t i o n of  evaluated.  panelists  independent  availability  to  scales,  rated  studied,  i n Figure  and  trained  rating  texture profile  defined was  on  standard  standard rating  Properly ria  and  the  The  adhesiveness,  observed  phase,  discussion.  complete  determined score  the  characteristics  a  was  sensory  could  group  acteristics oped  The  with  samples  through  residual  and  observed.  included  chewiness,  mechanical  viscosity,  i n i t i a l l y  characteristics  mechanical  personnel.  the  brittleness,  gumminess,  the  the  mastication.  Panel of  of  were d e s c r i b e d .  included  m a s t i c a t o r y phase,  geometric  third  hardness,  phases  complied method;  individual  -21-  with f i r s t  the the  two  crite-  data  ob-  observation,  and  Texture  P r o f i l e Score  Product:  Date: Time:  1.  I n i t i a l ( p e r c e i v e d on f i r s t a) Mechanical Hardness - Fracturability - Viscosity b) Geometric c) Other  2.  Masticatory (perceived during a) Mechanical - Gumminess Chewiness - Adhesiveness b) Geometric c) Other c h a r a c t e r i s t i c s  3)  Residual - Rate of breakdown - Type o f breakdown - Moisture absorption - Mouth c o a t i n g  Figure  1.2.  Sheet  Texture  bite)  chewing)  p r o f i l e score sheet  -22-  (Bourne,  1982).  second, panels  the held  This vided and  at  different  method  of  means o f  sensory  and  verified  by  other  places.  texture  profile  analysis  pro-  obtain descriptive  q u a n t i t a t i v e sensory  data  on  characteristics  products.  It  to  the  textural  advantage  of  food  product  or  through  rigidly  defined  and  nomenclature. the  had  any  objectivity  One  limitation  v a r i a b l e degree  of  the  flexibility,  textural  of  with  characteristic,  points  the  of  of  method  panelists skill  reference  was  related  when  applying  method. 1.4.  U l t r a s t r u c t u r e of 1.4.1.  tion and  Although  there  between  fish  Hallett, at  the  could  be  used  species.  fish, that  and  as  are  found  exist  of  muscle  differences  muscle  a  and  found  model  et  muscle  described  which  muscle  that  of  between  the  arrangement  the  relationship  of of  similar  fish  muscle nuclei  the  that the  to  of  each  skeletal  and  gross  and  -23-  the  organiza-  animals, enough  Bremner similari-  mammalian of  muscle  many  fish  the u l t r a s t r u c t u r e  fresh  and  salt  other  and  similar  muscle.  terrestrial  fibrils,  salmon  muscle  compared  species  mammalian  chum  were  level  describe  eight  in  there  a l . , (1986)  them for  to  of  terrestrial  ultrastructural  Shindo  skeletal  the  Structure  (1985)  ties  of  times  and  researchers  and  the  repeatable,  food  application  to  were  helping  food  a  results  mitochondria  to  Differences  animals  amount  water  of to  included sarcoplasm,  other  muscle  components, reticulum The and  (Bishop gross  Bremner,  which  each  other  from  of the  double fish  with  both  body, of  Within  each  proximately the is the  composed interior  separated  to  and  connective  tissue.  f i ttogether  t o each  Each  hundreds  of  the  basement  (Howgate,  units  non-branching,  of  elongated,  cells,  i s a  depicts  myocommata i n -  forces. take  The  the  shape  surface  axis  their  single  are  an  Myofibrils and  f o r the  the on  are  from  called  1.3  of  sides.  angles  to  cell, most  enveloped by  an  structures  of and the  diffuse  the  close  and  inner  outer are  a  ap-  occupy  of  of  run  environment  and  body.  long  composed  cylindrical  -24-  sheets  at varying  (plasmalemma)  the  the  (myomeres)  Myofibres  1979).  of  muscle  the  myofibres which  membrane  membrane  c o n t r a c t i l e  but  e x t r a c e l l u l a r  double  at  fibres  myomere  fibres.  semi-permeable layer  along  resting  other,  sheets.  from a  muscle  of  responsible  skeleton  appearance  sections  the  The  Hallett  separated  contractile  axial  by  Figure  were  which  muscle  sarcolemma,  and  muscle's the  sides were  1988).  the  sarcoplasmic  sheets  collagenous  of  of  both  myotomes,  sides  parallel  long  Bremner,  give  the  described  of on  transverse  myotome,  of  was  tail  called  shaped"  myocommatal  fish  consisting  the  and  "W-  of  head  of  1967).  t h e myotomes,  of  cones  series  as  of this  transmission on  Odense,  thin  (Hallett  terconnected  arrangement  blocks  by  structure  myotomes  and  (1988)  muscle  myocommata  the  structure  extended  Adjacent  the  and  basic  packed, of  about  Figure  1.3.  Appearance of musculature of a t y p i c a l fish. White spaces represent the p a r a l l e l muscle c e l l s , " d i v i d i n g " l i n e s r e p r e s e n t the myocommata w h i c h s e p a r a t e t h e myomeres. (Love, 1980).  -25-  one  micron  muscle  i n  diameter,  tubules,  the  the  cell's  between  function  either  scribed and  was  and  the  of  the  revealed  a  which  was  the result  thick  and  thin  structures,  of  (Dubowitz,  1985).  ference  i n as  the  the  of  s t r i a t e d  relative  compared  to  were  of  e t a l . (1986) width  of  mammalian  -26-  I,  the  de-  fibre, system, gly-  describes  of  a muscle  the  and  dark  fibre bands,  arrangement  the myofibril.  proteins,  pattern  lies  cell.  light  sarcomeres,  during  apparatus,  1.4  section  made u p  cell  reticulum  overlapping  different  Shindo,  Golgi  of a muscle  which  from  membrane  Figure  regular  bands.  s a r c o p l a s m was  tubule  of  A  muscle  cytoplasm of  filaments,  pattern  filaments  t e r i s t i c  the  of  myofibrils  the  of  sarcoplasmic  longitudinal  of a  the of  reticulum  the  T-system.  impulses  The  of  invagination  centres  electrical  droplets.  repetitious  alternation  the  as  components of a  as  network  the  surrounded  T-tubules.  (1985)  lipid  Observation  regular  an  The  intermediate  free  repetitious  T-system  sarcoplasmic  of  subcellular  muscle  length  lace-like and  mitochondria,  microtubules, cogen,  conduct  Dubowitz,  contains  major  the  reticulum,  and  relaxation.  side  by  to  i s a  which  Z-lines  the  and  the  membrane  the  into  contraction  myofibrils  described  plasma  membranes  on  the  sarcoplasmic  (1973)  midway Its  extend  fibre.  Surrounding  Peter  which  a  These  result  and  of  muscle.  the  were  charac-  skeletal  muscle  found a marked and  of  A  bands In  of  both  diffish white  Sarcolemma Nuclei, ribosomes  T-transverse l— system Intermyofibrillary -tubular complex  Myofibrils  1  —  Sarcoplasmic reticulum  Contractile proteins of myofilaments i . e . myosin, -actin, tropomyosin, troponin, actinin, M-proteins Mitochondria Lysosomes Golgi apparatus Glycogen p a r t i c l e s , l i p i d bodies S o l u b l e p r o t e i n s and enzymes o f carbohydrate metabolism  Figure  1.4.  Major s u b c e l l u l a r muscle. (Peter,  components 1973)  -27-  of  myotomal  and  dark  than  muscle,  that  muscles. finding  was  during  of  and  fibre  II  the  at  a  muscle, of  These  humans,  sarcomere  developed more  muscle were  than  the  had  a  1964;  than  and  muscle,  and  occurred  fibres  be  striated  by  fewer  muscle.  of  contractile  along  tissue  were in  of  tended  -28-  the  called to  swim-  surface  or  I  fibres  with  a  longer  fatigue  were  and  r i c h l y  with  well  contract  (Franzini-Armstrong section  red  type  mitochondria  cross  of  mid-line,  dark,  They  to  to  units,  during  fish's  diameter,  f i b r e s .  A  At  similar  With  constructed  energy  the  large  and  number  a l . , 1981).  mass  similar  be  muscle  1973).  to  glycogen,  of  The  coloured  to  number  and  pink  thought  smaller white  teleost.  characterized  of  production  large  Peter,  this  histologically  thought  was  type  et  or  less  coloured  white  that  which  marine  was  i n a band  inner cristae,  slowly  Porter,  brown  shrinkage  less  s t r i a t e d  however,  a l l fish  brown,  possible  lying  was  mammalian  in a  tissue  i t was  (Dunn  and  vascularized,  light  second  largest  speeds  strip  muscle.  the  bands  d e s c r i b e d two  muscle  This  for anaerobic  high  white  was  in  the  geared  ming  a  capillaries,  facilitate  to  sarcoplasm,  than  f o r  I  specimens.  i n humans,  less  the  feature of  (1967)  was  of  convinced  due  striated  fibres  mitochondria  and  the  muscle.  size,  blood  of  tissue  white  type  not  be  Odense  types this  called  few  i t might  preparation of  distinct  to  were  characteristic  that  width  i l l u s t r a t e d  workers  a  Bishop  bulk  relative  usually The  suggested  the  of  the  and  white  muscle or  fibres  nearly  angular, fibres. metric  revealed  that  the  circular  i n the  or  asymmetric  more  Dark at  muscle  any  centre  fibres  portion  m y o f i b r i l s were  of  of  muscle  at  were  the  fibres,  but  periphery  polygonal,  i t s cross  elliptical,  of  and  section  rectmuscle  less  (Shindo  asym-  et a l . ,  1986). Proctor fibre  type  trutta, the  et  called  brown  red  a l . , (1980)  and  trout. white  character  between  eter,  were  and  swimming  pink  fibres The  thought  to  distinguish  the  and  Adams,  muscle  distribution tissue,  presence  Pathological myofibrils,  or  the  sarcolemma  muscle  were  a  small  role  third  of  Salmo  situated  intermediate and  a  in  red  during  between  size  fibre  and diam-  intermediate  degeneration  (1985)  the  in  used  of  absence found  of  supporting  often  occurred  muscle  fibre  appear  to  the  (Peter,  -29-  the  or  in  other  character and  supand  factors. affect  (Dubowitz, the  to  lesions,  b i o p s i e s may  breaks  the  1985).  continuity  of  necrotizing injuries,  but  cell.  the  irregular,  volume  of  membranes  with  criteria  the  the  i n muscle  of  included  genetic,  i n t e r r u p t i o n s or  fibre  number  functional effect  their  may  a  Factors  changes  necessarily lethal  cells  decrease  the  changes  example,  not  have  diseases.  of  or  For  were  musculature  fibres  were  of  speeds.  Kakwas  porting  presence  the  large white,  1.4.2. M u s c l e  and  the  in  pink  fibres,  the  noted  The  surface  possibly 1973).  a  of  result  Folding  of  of  a  the  sarcolemma  is  split  away  from  folds  in  the  (Dubowitz,  1985).  departs  fibre  size  of  in  ing),  biopsies  severe  may  be  material.  such  as  abnormality  in  also  even  one  and  normal  branching,  or  or  by  an  of and  A  or  amor-  abnormalities band  loss  broadening  appearance),  muscle  characteristic  loss,  streaming,  the  Pathological  replaced  I  of  alterations  necrosis,  include  i r r e g u l a r i t i e s (zigzag  of  splitting,  lost  may  was  alteration  included  Selective  region  alterations  It  may  extensive  appearance".  distribution, In  or  common  because  "classical  membrane  creating  Loss,  most  assess,  muscle  structure  particular  Z-line  the  and  granular  a  to  myofibrils.  myofilament phous  from  observed  of  space.  muscle  changes  basement  membrane,  the  often  loss  plasma  the  considered  difficult  the  the  and  was  most  changes  common,  extracellular  myofilaments diseased  also  and  (thicken-  complete  disin-  tegration . Kakwas  and  fibres  into  mental  necrosis  many  Adams,  three  necrotic  in  tron  which  was  used  were  often  micrograph  changes, the  which  groups.  included  categorized  The there  sarcomeres.  disfigurement, lesions  (1985)  was  The  to  a  group  second  difficult  in  The  to  organelles.  -30-  of  types  and  muscle  a  seg-  few  or  myofibrillar  of  distinguish group,  in  called  group, series  third  fibre  was  clustering  classify a  artifacts. changes  first  changes  subcellular from  elec-  morphological alterations  of  Based  on  mammalian  that  the  tion  contributes  which  physical  are  fatigue  of  curring  during  had  process,  and  following  Friden  micrographs  of  bances  the  were  regular  loss  was  found  in  fibre  of  the  that  second  third,  of  related  the  wavy  the  repetitious  stage  associated  with  extreme  revealed  electron  found  pattern  and  a  of  of  to an  the  point  increase  -31-  of in  the  in  fatigue  man.  also  disturof  these  focal  Z-bands. and  an  Gaps appar-  observed.  acid  the  streaming,  exhaustion the  of  Light  focal  marked  Z-bands,  were  muscle  myofibrillar  myofibril  the  oc-  Myofilament  level,  r e s u l t of  myofilaments  exercise  necrosis  be  star-  changes  p a t t e r n  microscopic  disruption  lattice thick  to  of  conditions  observed  band  contrac-  in rat  of  sections  pro-  contractions.  last  exercise  the  system.  changes  eccentric  the  and  of  hormonal  reproductive  (1983)  i n  effects  the  a l . ,  Factors  energy  disoriented.  at  migra-  first,  types  the  the  to  and  et  total  of  gathering  believe  chum.  changes  bursts  c r o s s - s t r i a t e d  disturbances  ent  and  semi-thin  At  the  force  intense  myofilament.  and  rapid  prolonged,  only  damage  are  to  spawning  of  f o r  a l . , (1985) o b s e r v e d  was  use.  broadening,  low  observed  muscle  of  both  appeared  was  during  deterioration  salmon  development  sarcomere  occurs  a l . , 1985),  undergone  dissolution  in  with  et  et  i t i s reasonable  r e s p o n s i b l e  migration,  Belcastro which  muscle  repetitious,  during  that  spawning  (Belcastro  vation  The  to  associated  longed, tions  stress  l i k e l y  ultrastructure  studies,  It  resulted  hydrolytic  activity was  of  known,  were  muscle however  liberated  sponsible  f o r  tigue. number  of  the  histopathological  to  lysosomal  Pale non  whereby  post a  mortem  low  pH  investigated  utes  post  mortem,  were  served  i n the  et  a l . (1981)  in  t h e PSE  down w i t h  No  also  muscle loss  been  re-  degradation  of  a l . , (1987) muscle  and  an  of  apparent  i n the  muscle  and  i n the  studied  the use  i n  i n the were  i t was  Canada and f a -  increase  were  (PSE)  pork  which  the  muscle.  correlated noted  muscle  i s a well  temperature between  Cassens  et  that  mimicked  24  the  of  o b s e r v e d an after  and  death,  of material  mortis  i s s t i l l  normal  fibre  and  including  -32-  of  to occur  at  high. PSE  was  40  appeared were  muscle. of  ob-  Cloke  disruption  myofibrillar  Z-lines.  min-  disrupted,  bundles  pork  The  muscle  Within  unusual degree  at the  rate  in myofibrils  normal  phenome-  accelerated  material  alteration hours  known  a l . (1963).  together,  marked  an  causes r i g o r  sarcoplasmic  first  have  excess muscle  animals exhibit  "clumped"  disconnected.  may  the  et  of  changes  carcass  by  of  breast  time  differences  were  and  animal myopathies.  glycolyis,  ultrastructural  I-bands  and  the  fibres,  a c t i v i t y ,  exudative  while  enzymes  changes  necrotic  stressed  lysosomal  bodies  enzyme  soft  suggested that  streaming,  lysosome-like  o f human  changes  George  a  Z-line  The  symptoms  f o r these  i n the  migration,  flight-induced  mechanism  i n i t i a t i o n  changes  Excess  No  damaged  material.  ultrastructural after  i t was  i n the  m y o f i b r i l l a r  goose  enzymes.  break-  1.4.3. The for  energy  survival,  first  by  of  the  myotomal  (saithe) loss and  in  and  body  area.  in  had  myofibril  in  becomes d e p l e t e d , the  white  require  is  the  obtained  liver  there  muscle  atrophied  starvation.  fibres  were  tained  their  by  M-line of  the  is  and  exten-  for  energy  62%  the  50%,  and  25%.  proteins, myofibril  fibres,  degraded,  the  on  flatfish.  In the  the In  1981,  the  was  area  -33-  of  muscle  of  the  a  of  myotomal  filaments  sarcomeres  white  of  the  preferential  In  myofibrils  severely whole  s t i l l  main-  the  skeletal  muscle  at  and  studied  of  the After  density  thick  Johnston  of  cell.  observed.  the  the  volume  volume  in  stores, twitch  muscle  of  percent  cross-sectional  addition,  and  L.  (red)  Atrophy  ultrastructure Atrophy  f i f t y  slow  the  virens  glycogen  myofibril  myosin  however  alignment.  A  and  of  ultrastructure  Pollachius  starvation, to  by  the  tissue  cross-sectional  decreased of  in  observed.  in  84  fish,  (white)  of  from  mean  loss  was  days  starvation  marine  80%  decrease  muscle  fibres  of  of  decreased had  studied  period  the  74  (1985) fat  fast  the  periphery  a  stores  low  of  a  degeneration  from  lipid  salmon  development  the  an  reduced  months,  fects  gonad  from  weight,  After  myofibril  of  a  muscle  resulted  the  supply  Johnston  muscle  atrophy  muscle  available  proteins  during  myotomal  3  and  maturing  1980).  Beardall of  non-feeding,  migration,  When t h i s  use  (Love,  that  utilizing  muscles. sive  Starvation  ef-  muscle  fibres  was  associated volume  with  occupied  degeneration thin  a  thin  whereby s t i l l  were  supported  after  s t i l l  14 m o n t h s  h a d some Beardall  of  had  diameter thick  that  used  cells  an  that  of starvation, fish,  thick  a  and  gradual  mechanism  source This  studied  while  hypothe-  histological  Cyprinus  the parallel  although  In se-  this  energy  of the fish  and t h e  lysosome-like  functions. who  effect  fish.  provided  as  (1980)  this  diameter,  suggested  of the  carpio.  orientation  largely  destroyed,  recognizable myofibrils. and  muscle  myofibrils  eral  be  peripheral  disorganized  diameter  Love  i n these  starvation-induced white  by  i n t h e muscle  myofilaments  the  could  was  multi-membrane  I t was  i n myofibrillar  fractional  i n starved  fibres,  and numerous  and  the unraveling  suggested  m a i n t a i n i n g some c o n t r a c t i l e  changes  of  caused  function  white  observed.  common  i n myofibrillar  of contractile  myofibrils  was  Even  Also  Johnston  decrease  filaments,  reduction  sis  lattice.  degenerating  vesicles  i n the diameter  by m y o f i b r i l s .  partial  maintenance verely  decrease  o f Z-disks, which  filament  caused  a  a  Johnston  (1985)  degeneration marine normal  stage,  thick  appearance.  aments  and Z - d i s c  tion,  the myofibril  were  (myosin)  myofilaments  that  f l a t f i s h .  of the myofibrils filaments  listed  were  intact. had  could At  absent,  stages  found  stage  of  i n the  one,  In t h e second  the  stage,  and t h e p e r i p h -  absent. leaving  In the fourth  -34-  be  reduced,  were  been  four  stage  completely,  In the the thin of or  third f i l -  degeneral a r g e l y  disrupted,  and  Haard catch was  of  phagocytosed.  (1987)  the  American  unsuitable  which  was  normal  observed  for  market  colour  fillets  of  cent  appearance.  in  these  and  translucent,  and  indefinite  smooth,  and  in  and  and  in  jellied  89%,  and  The  myotomes  were  of  levels  and  actin  amination muscle section of  of  the  fixation  and  thought  be  there  l e s s muscle  the  looser  Andrews, due  and  to  from  protein  starvation  the  18  -35-  wide  was  loose,  each  %  to  was other  of  78%, 9%.  showed  reduced  spaces  the  jellied  ex-  between  area.  possibly  to Gel  Histological  unit  the  content  normal  per  the  was  water  Long  a  result  jellied  tissue  condition  emaciation of  opales-  fillets  from  large  of  and  the  from  waviness,  This  the  analysis  1987).  structure  muscle,  itself  the  material  1956).  partly to  also  that  were  fibres exhibited  on  (Templeman  maturation,  and  that  (Haard,  the  septa  away  reduced  to  glossy,  myofibrillar protein  myosin  revealed  fibres,  to  the  was  the  Proximate  increased  protein  of  of  the  condition  contrast  face  revealed  of  platessoides)  tissue  pulled  40%  In  tissue  cut  1956).  fillets  electrophoresis of  connective  fillets  to  jellied  texture  connective  the  a  jelly-like,  nature.  crude  of  consumers.  were  20  (Hippoglossoides  firm  The  jellied  the  to  Andrews,  normal  between  because  and  fish  and  the  (Templeman  the  plaice  unacceptable  greyish  that  fish.  during  was  sexual  1.5.  Proteinase 1.5.1.  Dean as  size  brane,  they  erally  had  were  pH  cell  peptides  and  component amino  nucleosides Although  and an  during  direct  involvement  radation many  of  other  there  calcium Doyle, cated  o v e r a l l  periods of  Besides  lysosomes  of  rapid  lysosomal  the  although  some  i n c l u d e d  phospholipases, of  degraded  these For  to  small  free  fatty  nucleic  acids  to  and  enzymes  an  i n  the  activity  protein enzymes  proteins  neutral  However, as  gen-  lipids  of catheptic  i n the  has been r e the  i n the i n i t i a l  deg-  i n  serine  site  There  system,  proteinases,  (CANP)  are  myofibrillar  lysosomal  circumstantial evidence  intracellular  of  degradation,  implicated  proteinases  -36-  number  i s unproven.  alkaline proteinases,  activated 1978).  were  increase  p r o t e o l y t i c enzymes  are also  which  phosphates.  myofibrillar  breakdown.  enzymes  macromolecules.  to monosaccharides,  a n d t h e amount  ported  mem-  function  o f complex  of  thick  nucleases,  complex  vesicles  enzymes  The  system  single  range,  These  proteins  acids,  a  of  acid  sulphatases.  carbohydrates  lysosomes,  pH.  the digestion  by  array  i n the  n e u t r a l  and  was  example,  to  diverse  glycosidases,  phosphatases, organelles  a  the lysosomal  membrane-limited  Surrounded  optimum  at  proteinases,  (1976) d e s c r i b e d  shape.  contained a  system  intracellular  and  a c t i v e  acids,  Lysosomal  and B a r r e t t ,  sedimentable  varying  activity  (Segal  and and  which  impli-  f o r protein  break-  down  included  studies with and  a  i n which  number  of  of lysosomal  turnover form,  is  i t suptake by  degradative  to  that  a  process  half-life  are related,  completed minal  by  within  both  intra  through lysosomal The  than  (Dean of  and  than  which  and  interaction  within  -37-  and  protein  solid  between  the i s a  polypeptide pro-  weight  point  and  are de-  basic  ones.  i s initiated a  peptide,  newly  by  and i s  exposed  ter-  1976).  specific  other  there  proteins or  i t  of  weight  isoelectric  release  cell,  molecular  system  turnover,  i n the  molecular  neutral  and B a r r e t t ,  extracellular  of the  acidic  bonds  and i t s  features  small  lysosomal  protein  membrane  large  the  cleave  system,  First,  weight  protein  susceptible  eukaryotic  turnover.  f o r example  the  which  changes  precise  the size  of  to a  distinct  protein's  exopeptidases  residues Control  the  r a p i d l y  endopeptidases  normal  a r e two  rapidly  Second,  more  In a  f o r example  more  the process  digestive  the molecular  proteins.  Proteolysis  or with  of a protein  of protein  between  breakdown  graded  described  lysosomal  there  i t shalf-life,  teins  correlated  turnover  proteolysis.  relationship and  Protein  e t a l . (1978)  accepted  pathological  of proteolysis  proteinases,  as t h e transformation  breakdown  and  lysosomes.  1.5.2. Segal  physiological  the overall rates  the activity number  of  the  degradation  was  enzyme  brought  binding  supports  t h e membrane  of  about of  the  (Dean,  1978).  and t h e  enzyme  was  therefore  important  conversion  of  neutral  optimum  pH  curred at  pH  most 4.5  To  action  phobic  were  parts  or  strate cise  ditions within  not  of  pH  was  or  as  degrade  example,  from  a  alkaline bound  to  form  pH a  mechanism  the  with  a  optimum  oc-  membrane,  but  between  that  i n very  of  connectin,  the  on  heavy  chain  to  which  surrounds  in  the  with  high  the  required  2.8-4.0,  source  of  function  nebulin,  filamin,  neutral  myofibril  -38-  at  cleave of  the  sub-  although  pre-  enzyme  for acidic  could  only  conoccur  locations.  proteins (>  and  caused  200K)  pro-  and  myosin  heavy  proteinases  (CANP)  could  weight  extent,  adjacent  peptides  the  weight  of  hydro-  to  extracellular  molecular  in-  points  against  myofibrillar  molecular same  pH  to  i n more  unable  requirement  vivo  Its  preferably  was  and  restricted D  D.  activity  the This  activated high  enzyme  the  lysosomal  endopeptidase  clustered  chains,  Maximum  depended  major  cathepsin  This  of  i t i s necessary A  substrates,  cathepsin  Calcium  is  polypeptide  weight  degradation  such  and  strongly  chains.  suggested  extensive  chain.  be  residues.  cells,  an  when  role  ( B a r r e t t , 1978).  Incubation  teins  to  the  side  optimum  with  6.5  lysosome  hemoglobin  substrate  pH  the  found of  more  form  characteristics.  the  molecular  five  For  diphosphatase  i n catabolic processes,  in  aromatic  low  a  i t s activity.  solubilized.  their  only  cleavage  to  to  1,6  rapidly at  when  vestigate found  fructose  understand  enzyme  to  proteins  however the  or  desmin,  Z-line  was  the a  myosin protein  removed.  The  incubation  with  electrophoretic Bird  abled was  acted  t o then  further  i n a cooperative  to that  that  changes  not established  by c a t h e p s i n  similar  suggested  conformational  CANP  followed  pattern  e t a l . , (1978)  stantial  it  CANP  D produced  of cathepsin  cathepsin  D  caused  i n the molecule,  degrade  conclusively fashion  that  suben-  However,  t h e two  t o degrade  D.  which  the molecule.  an  enzymes  myofibrillar  pro-  teins . Another  endopeptidase  myofibrillar cathepsin pH  breakdown  B.  Maximum  6.0, a l t h o u g h  strates  (Barrett, with  natured ments The  t o an  a  from  lecular  weight  tant  incubation  and myosin  myosin  group  10,000-50,000. 150,000  the thiol most  were  found  of the  size was  of less  less  sub-  acidity  a t pH  5.2  of defrag-  than  30,000.  extensive,  and r e -  of polypeptides  also  B  was a t  some  polypeptide  In addition,  was  with  the degradation  smaller  of  proteinase  substrates  effect  caused  molecular  of native  t o the formation  of cathepsin  to progressively  heterogeneous  weights  with  pH o p t i m u m s  ultimate  degradation  leased  The  actin  myosin  was  of the denaturing  1978).  purified  products activity  lower  because  o f importance  produced  a  with  molecular  compound which  was  of  mo-  resis-  to further proteolysis. 1.5.3. Millward  systems  e t a l . , (1978) f o u n d  during  increases  Starvation  s t a r v a t i o n were  i n protein  that  implicated  breakdown.  -39-  t h e muscle  lysosomal  i n the catabolic  F o r example,  during  nutrient 30-150% an  deprivation,  in skeletal  increase  in  starvation suggested mentally the  was  the  that but  the the  Thus  the  The  activity system  but  Other  explanations  normally  The  was  of  reduced. of  the  cell  normal  A l l types  of  The  of  normal  lysosomal  normal been  proteins thus  the  put  protein uptake  was  and  net  the  was  same,  enhanced.  degradative of  be  became  suggestion  system  to  accepted  half-life,  second  p r o t e i n s would  that  suggestion  p r o t e i n s remained  degradative  involved the  the  and  fundaand  have  who  susceptible  stable  size  during  was  first  more  The  (1978)  acceleration  became  molecular  half-life  stituents.  Walker,  p r o t e i n a s e - s e n s i t i v e , and  between  and  degradation  degradation,  catabolism.  proteins  susceptibility  chinery,  hanced  enhanced  attack.  correlation and  an  showed  lysosomes.  catabolism  protein  simply  enhanced  studies  the  and  protein  normal  not  c e l l  preferentially  of  Dice  Several biochemical  proteolytic  charge  from  was  for this  that  by  increased  was  morphological  fragility  described  different  proteolysis.  and  and  catabolism  intracellular protein  the  mechanism  forward  of  were that  muscle,  size  characteristics  protein  ma-  cellular  degraded  con-  at  en-  proportional rates. explanations  hypothesized  to  include  proteinases.  Dice  and  for the  Walker,  non-lysosomal  proteinases  their  during  activity  the  enhanced  degradation  involvement 1978  in  muscle,  starvation.  -40-  observed  If  and these  of  non-lysosomal  the the  were  presence  of  increase  in  enzymes  were  to  use  small  normal point  and  correlation and  and  mechanism benefit a  be ity  from  tend  tageous  to  the of  Other  to  be  with  crucial  regulatory  en-  proteins,  because  disruption  of  organisms use  stable  least  the  acids for  these  have  also  remove  Z-bands,  C-protein.  been  and  However, disassembly cleavage  shown  enzyme  i t was of of  i n  (Asghar  degrade  This  the  to  thick  be  advan-  they  could  cell's  abil-  myosin  lysosomal  degrade  was  play  thin  pathway.  -41-  intact  and  degradation  of ex-  tropomyosin, actin  important  heavy  of  myofibrils,  degrade an  For  filaments prior  light Final  to  stages  1987).  complex,  unable to  possibility  i n i t i a l  Bhatti,  troponin  and  the  the  and  thought  proteinases.  v i a the  suggested  p r o t e i n a s e s  have  plished  amino  i t would  the  workers  myofibrillar  conditions  proteins,  CANP  specific  the  whatever  stable  ample,  the  starvation  that  the  degradation  in  isoelectric  reduced.  commented  Because  myofibrillar  myosin.  be  with  the  metabolically.  non-lysosoma1  and  s u b s t r a t e , then  increased production of  degrade  adapt  their  breakdown  would  (1978)  energy.  not  sacrificed to  weight  Walker  as  protein  occurring, during  source  zymes  proteins of  molecular  Dice  as  basic  to  chains was  and role the by  accom-  2. C H E M I C A L  2.1.  Introduction  The their  spawning  life  cycle  ronmental  and  stresses.  migration i n which  These  lengthy  fungal  disease,  elevated  sexual  maturation  quality. of  By  occur  The lows  to  1986a). such  as  lated  Idler snout  these  and  included  a  near  and  other  and  Bitners,  of  i n protein  and  of the fish  and  starvation,  Bitners  their  be  indices  such  and  (Ando  commercial points  accurately  wasting  of  physical  changes  and  weight,  and r e -  as  changes  i n  Other  the alimentary  and  and an  (1960)  i s commonly Hatano,  i n internal  t h e male  fol-  and  measured  thickness,  -42-  and  biochemical  approximately  maturity  (1958)  lipid  fresh  b a c t e r i a l and  and v i s c e r a l weight.  Both  to  assessed.  of  1960).  Idler  salt  may  stage  complete  i n  envi-  at different  the fish  significant alterations  loss  however,  changes  severe  The  of the animal,  skin  moisture,  from  upon  period  unaccustomed  migration,  impact  value  and Tsuyaki length,  and  state  to biochemical  protein,  enced  these  appearance  the  endure  e t a l . , 1965).  and i t seconomic  indicate  i s a  temperatures,  i n the fish  the physiological  used  water  water  the state  external  must  changes,  fresh  quantitating  salmon  t h e movement  (McBride  the migration,  described,  chum  the fish  include  an o f t e n  which  of  physiological  water,  changes  ANALYSIS  organs  female increase  reported  that  fish  f a t ,  changes tract, (Idler experi-  i n moisture the  females  had  22%  more  f a t  loss  and  25%  more  protein  loss  in  the  muscle. Hatano of  fall  The  creased  object  of  this  was  moisture  on  i n the muscle Materials 2.2.1.  Fish  were  except  Chilliwack were heavy  gauge  pearance  to  (1986).  A  The  10  grades  over  fish  Hatchery  were set  tenth  were  A  as  to D  2  first  divided  16%,  the  assess the  the  fish  appearance. i n crude  year  from  effi-  into  The  protein,  four  second f a t , and  chum  salmon.  period.  In  a  which  bags,  local  were  (Chilliwack,  washed,  first  packing  plant  collected  B.C.).  dressed, and  the  A l l the  packaged  stored  at  from  in  the fish  double  -30°C.  Grading graded of  9  grade  grades  a  obtained  polyethylene  a  to  in  Methods  sampled,  fish  20  salmon  Sampling  2.2.2. The  from  the migrating  spawning  River  graded,  and  were  f o r the  which  chum  the moisture content i n -  to  changes  of  sampled  a l l fish  was  external  the  spawning  and  study  their  chemical composition  decreased  1.6%,  grading system  t o measure  2.2.  year,  to  80%.  based  object  8.5  the  (early)  content  to  a  compared  summer  72  of  grades  from  from  The  and  protein  dropped  ciency  a l . (1983)  (late)  Japan. lipid  et  comparing  pictures was  then  grade  by  added  reduced  1,  grades  their  of  chum  to  include  to  four  E to G  external  composed  spawning  grades as  by  grade  by 2,  ap-  Anon. fish.  assigning grades  H  to  I as  as  grade  grade were  grade 4.  the  A  were  dicular  colour  black  bars  fish,  with  or  which  intensive,  green  or black,  ered  with  a  layer  of  very  dark  green,  skin  and  appearance  of  slime.  slime  the  four  2.2.3.  or  The the  total  method  tracted sample utes  from with  in a  added  Bligh the  25  mL  spark  twice  seconds. filter  of  to  The  paper  white  the  Dyer muscle  blender.  mixture  homogenate with  a  Buchner  noGrade  silver of  dark  skin  perpengreen  or  Grade  perpendicular  colour  was  thick  salmon  used  and  cov-  fish,  colour, 2.1  lipid  had  very  thick  illustrates in this  the  study.  analysis by  (1959). by 50 25  and  was  no  fish.  form  to  Figure  determined  c h l o r o f o r m and free  and  spawning  skin  to  surface.  skin 4,  used  analysis  Total  was and  of  the  dorsal  Grade  grades  Proximate  lipid  the  cover.  to  yellow  sheen,  red  I,  silver-brights,  i n the  distinct, and  1,  uniform  light  than  criteria  on  the  changes  had  heavy  marks  ventral  of dark  the  silvery  of  were  the  black  a  loss  colour  from  of  more m a t u r e  Grade  water  bars  a  fish  follows.  showed  distinct  i n colour,  dark  as  changes  intermediates,  J " , any  description  i n colour  colour  and  "grade  brief  fish  colour,  3,  and  blue-black  ticeable 2,  3,  slightly The  lipid  homogenizing mL mL  methanol of  25  was g  ex-  of  the  f o r two  min-  c h l o r o f o r m was  then  homogenized  each  time  filtered  through  funnel.  Additional  -44-  modifying  Whatman  for  30  No.  1  lipid  was  Grade  Figure 2.1.  1  -  Silver bright  Grade  3  -  Dark  Grade 4  -  Spawning  I l l u s t r a t i o n o f t h e appearance o f t h e f o u r grades of chum salmon used i n t h e s t u d y . -45-  recovered tissue. ated  by  re-horaogenizing  The  filtrate  cylinder,  sured,  and  the  the  tent  was  form  mixture  found  by  removed  using  and  drying  the  granular was  of  tared a  stored  [Total  in  the  W VI V2  to  round  a  dark  in  until  they  were  W  *  a  moisture  triplicate at for  70°C 1  5  for  hour  g  of The  ensure  crude  a  method  using  Approximately digested trated  in  of  protein  the  30  sulfuric  g  of  mL  chloro-  chloroform  bath  at  45°C,  nitrogen.  The  desiccator  weighed.  samples  Total  with lipid  acid  and  muscle were  Protein of  chum  Kjeldahl  was  found in  a  by  drying  vacuum  weighed  and  oven  re-dried  weight.  content  dried  fish  chum  for micro-Kjeldahl 9  con-  analysis  the  constant The  vacuum  lipid  V1/V2]  content  hours.  to  of  mea-  the  The  water  was  weight of l i p i d i n a l i q u o t volume of c h l o r o f o r m l a y e r volume of a l i q u o t  samples  6  of  gradu-  equation: M o i s t u r e The  in a  The  flask. a  residual  layer  5 mL  stream  the  =  = = =  dryness  in  the  settle  removed.  bottom  with  to  chloroform  was  following  lipid  Where:  the  rotoevaporator  gel  using  re-filtering  allowed  layer  residue  silica  found  volume  evaporating  in a  were  then  methanol  was  flasks  was  and  determination the  -46-  muscle  digestions muscle  digestion  HgO  chum  (Anon.,  (0.5-2.5 flasks  catalyst.  was  The  mg  with  found 1970). N)  was  concen-  mixture  was  heated  for  15  to  30  added  as  an  seconds  ide  was  the  organic material.  solution  was  m o n i a  cent  oxidant  diluted  c o n t e n t  protein  constant  i n 25  a  1  mL  speed  the mL  t h e  with  was  to  Once  of  colourimetrically  and  up  2  astaxanthin  lipid  extracted  grade  filter  475  nm  by  the  am-  II.  the  The  ppm  N  per-  by  the  The  assuming (Skrede  and  and  filtering  HunterLab  Labscan  ates  Inc.,  Lab. inch  proximate  L,  the  6000  was  dimensions  a,  cm  X  in  No. 1  UV  (Shimadzu  160  Corp.,  c a r o t e n o i d was (1%)  of  cm  cal-  1900  at  1986). Lab  and  b  the Raw  1  in  Whatman  Shimadzu  measurements in  the  spectra  Spectrocolourimeter in  (1959)  measured  a  the  reflectance  used. 2  were  coefficient  Hunter  Virginia)  aperture  i t through  using  Storebakken,  resolubilizing  method  Spectrophotometer  extinction  from  by  Dyer  readings  cuvettes  parameters  calculated  and  concentration of  an  measurement  measured  Bligh  Recording  Japan).  Colour  1/4  and  the  d e t e r m i n e d  Autoanalyzer  multiplying  of  completed,  was  the  were  was  s a m p l e s  was  Absorbance  quartz  U.V.-Visible  culated  content  acetone,  paper.  length  Kyoto,  decomposition  flasks,  by  perox-  Colour  The  path  the  volumetric A s t a x a n t h i n  reagent  hydrogen  6.25. 2.2.4.  the  30%  digestion  Techicon  determined  of  cm  -47-  region muscle X  0.5  of  raw  chum  recorded (Hunter 400-700  segments cm  flesh  were  of  by  a  Associnm.  A  the  ap-  presented  to  the  instrument  on g l a s s  tissue  from  ferent  locations,  a,  each  fish  slips.  were measured  and t h e values  and b readings  2.3.  cover  f o r each  Results  Triplicate three  were  times  samples  each  averaged  at  of  dif-  to obtain  L,  sample.  and D i s c u s s i o n  2.3.1. Astaxanthin content The formed  grading the basis  w e r e made it  was  the  about  they  t o confirm  corresponded  they  maturation  t h e muscle  ries  v i a t h e serum. i n chum,  loss The  mean  astaxanthin  carotenoids  Analysis  of variance  termined  that  there  between  at least  between  t h e grades  are of dietary i n t h e muscle, colour.  from  the fish  ova-  carotenoid  when  following  randomized difference  one o f t h e grades.  Multiple  with  -48-  single  of the  i n Table  was a s i g n i f i c a n t  made  and  the analysis  are given  of the completely  were  With  are mobilized  i s t h e main measured  as the  (Ando e t a l . , 1985).  obtained of  appearance o f  t o t h e integument  Astaxanthin  reason  occurred  red-pink  the carotenoids  of the fish  For this  deposited  f o r i t s typical  values  that  because i t  a l l conclusions  the exterior  and i s t h e one o f t e n  content  which  changes  and t r a n s p o r t e d  i n colour  important  grades.  are mainly  however,  from  the  that  t o other  are responsible  found  from  In salmonids,  and because  sexual  very  d i f f e r e n c e s between  matured.  origin  was  o f comparison  important  fish  fish  of the fish  degree  2.1.  design (a <  de-  0.05)  comparisons of  freedom  Table  2.1.  Mean v a l u e s f o r astaxanthin content d o r s a l m u s c l e o f chum s a l m o n (mg/kg ) .  Grade  1 2 3  n  i n the  Astaxanthin (mg/kg m u s c l e ) 3  1  5  6.53  (2.6)  a  2  6  6.07  (3.3)  ab  3  10  2.78  (1.8)  be  4  6  1.08  (0.7)  c  Mean ( s t a n d a r d d e v i a t i o n ) , "n" d e n o t e s sample s i z e . Different letters signify difference.  -49-  a  significant  contrasts  (Wilkinson,  sequential DiPonzio grades  1988)  rejective  and  0.05)  from  grads  1 and  4.  1987).  Grade  grade  4.  2  used  L,  and  a  existed by  of  variance  4.  The  and  'a'  extracted the  different  (a  diferent  creasing  grade,  <  rom  freedom  and The  different  4.  and  and  the  are  2,  grades ratios between  a  found  and  each  differences  with  2  to  be-  from  values  and  rejective  3 were d i f f e r e n t  'b'  /L,  the  with  3 and  and  comparison  increase  from  1  2.2.  differences  Multiple and  was  itself.  i n Table  that  following  decreased  muscle  shown  b.  were  1,  The  the  contrasts  values  grade  in  rigorous  colourimeter  indicated  a,  different  grade  4.  L,  without  Hunter  changes  grades  values  2 were  from  and  of  'L'  grade,  nificantly  from  values  detected  grades.  1 and  3  of  procedure  different  grade  and  different  significantly  was  colour  parameters  degree  increasing  grades  Lab  astaxanthin  measure  i n the  Bonferroni  grade  3 was  b v a l u e s were measured,  the  was  improved  (Holland  significantly  to prevent oxidation,  single  tween  the  to  Analysis  1  the  4.  Because  also  with  procedure  Grade  was  Grades  2.3.2. H u n t e r  procedures  corrected  Bonferroni  Copenhaver,  3  and  increasing 4,  grade  grade  decreased  were a/b  and  grade, 3  was  with  i n -  different  proved  to  from  be  except between  sig1  and  2. Examination astaxanthin  of  Tables  c o n t e n t was  2.1  and  2.2  revealed  roughly proportional  -50-  to  that 'a'  mean  values.  Table  Grade  2.2.  n  H u n t e r L a b mean v a l u e s chum s a l m o n .  L  a  value  1  6  40.02  (0.51)  a  2  7  35.82  (0.18)  3  11  41.59  4  11  47.08  f o r the dorsal  value  b  muscle o f  value  10.05  (0.07)  a  11.45  (0.16)  a  a  7.80  (0.13)  a  10.52  (1.46)  a  (0.41)  a  4.40  (0.09)  b  9.40  (0.08)  b  (0.48)  b  -1.30  (0.10)  c  7.67  (0.13)  b  3  "n" d e n o t e s sample s i z e . Mean ( s t a n d a r d e r r o r o f mean). Different letters signify a significant difference.  -51-  (P <  0.05)  A  more  detailed  correlation fourgrades Hunter for  a  and  were  the  and  The  was  performed  was  out  between  the  astaxanthin,  the  so  that  creased.  The  The ratio  as  i s not  a/L  i t was  in  reflected  astaxanthin, flected  the  had  commonly  colour, 'a'  the  a  used  the the  relative of  were  i n the  choice  in  r  in in  as  and  a/b  at  the  1%  values  a  de-  the  of  decrease  s p e c i f i c a l l y  value  in  this  measure  The  may  connective  protein  in  Although  case.  'L'  their  significant.  value.  this  a/b  shown a,  their  not  for  grade  by  and  are  carotenoid,  increase  contractile  to  literature  in  increase  tested  test  increased, L  a/L  astaxanthin,  largest  loss  were  correlated  and  the  logical  and  concentration  b  ratios  this  a/L,  grade  parameters  ratio  of  values  of  the  correlations  significantly negatively  level,  and  results  colour  carried  values  The the  relationship  concentration  colour  fish.  significance, 2.3.  b  the  which  salmon,  individual  Table  at  analysis of  L,  look  have  tissue muscle  re-  as  the  was  d i -  minished. The monly In the  colour  used  this  in  study  dorsal  grade  of  et  maturing  the  muscle  industry i t was  muscle  chum.  distinguish Aksnes  of  between  the  not  clearly  'a'  Hunter  a l l of  the  salmon,  -52-  loss  in  indicator and  value  however,  grades  except the  salar.  com-  maturity.  astaxanthin  differentiate  assessed  Salmo  i s an  i t s quality  that  a l . (1986) v i s u a l l y Atlantic  salmon  assess  found  did The  to  of  loss  between was one of  Changes  in each  able and  two.  colour in  to  in  colour  Table  2.3.  Independent  Results from the c o r r e l a t i o n grade, Hunter Lab values, content.  value  r  1  analysis between and astaxanthin  value  a/L  -0.796  **  astaxanthin  -0.746  **  a  -0.736  **  a/b  -0.750  **  b  -0.468  NS  L _  0.566  NS  2  ' n ' , t h e number o f s a m p l e s , was Significant a t t h e 1% l e v e l , NS a t t h e 5% l e v e l .  -53-  24. = not  2  significant  were  found  colour than  of  the  salmon,  the  i s  sulted  that  in  from  physical  from  chemical  the  in  the  dation  of  of  because of  the  method  of  are  so  An  The lipid  results  are  given  ance  was  ence  between for  differences significantly also  Grades  of in  1,  the  the  or  the  the  the  vivid  the  obtained, they  at  and  were of  A  a due  inconsis-  have  the  any  re-  occurred  time  process. do  found fish  not to  by  protein,  one-way  whether  f o r the  revealed  grades. <  or  during Because  cause be  a  oxi-  better  i t s colour.  different  -54-  there  three  was  < (P  1,  2,  grade  0.05) <  a  and vari-  differ-  parameters.  significant  from (P  moisture,  analysis of  Grades  0.05)  different  were  was  astaxanthin  could  of  analysis  2.4.  (P  value  this  for  This  grades  i s primarily  of  resolubilizing  grades  different  3  which  a n a l y s i s of  protein  the  f a t extraction process,  determine  fish  between  and  less  Hunter  carotenoid  grade  Table  significantly 2,  the  easily  to  crude  fish,  technique  Proximate  completed  results  thus  of d i s c r i m i n a t i o n  between  explanation  during  determining  lack  'a'  carotenoid.  carotenoid,  2.3.3.  the  the  the  oxidation of  the  process,  significantly  observe  isolation  losses  values  was  distinguishing  extraction, drying,  Hunter  was  to  content.  loss  maturation  fish.  redness  astaxanthin  tency  fish  especially  of  i n the  maturing  astaxanthin  measure to  the  late  i s interesting  the  the  occur  immature  It of  to  0.05)  (P and  4.  between from  <  The 0.05)  3  were  Moisture grades. grade  4.  Table  2.4.  Mean v a l u e s moisture and chum s a l m o n .  Grade  Protein  n  (% o n a w e t l i p i d content  Moisture  basis) of protein, of d o r s a l muscle o f  n  Lipid  n  6  1  17 .9  a ( 0 .96)  6  7 4 . 17 a ( 0 . 2 9 )  9  4 .9  2  19 .0 a ( 0 .04)  6  7 5 . 02 a ( 0 . 0 9 )  7  3 .8 a b ( 2 . 1 )  9  3  18 .3 a ( 0 .13)  11  7 5 . 58 a ( 0 . 2 7 )  11  4 .3 a b ( 2 . 3 )  6  4  15 .8 b ( 0 .45)  10  7 9 . 47 b ( 0 . 3 2 )  11  1 .0 b ( 0 . 9 2 )  4  Mean v a l u e s ( s t a n d a r d . e r r o r o f mean), " n " d e n o t e s s a m p l e s i z e o f mean. Different letters signify significant  -55-  a(2.0)  difference.  The  analysis  ence  (P  <  of 0.05)  however  only  other.  These  went  a  until that  in  the  ied  effect  nificant flesh  and  the that  value  ability sexual  of  as  of  imental  the  of  an  analysis  were of  the  same. on  quality  of  quality  and  of  indicator  f o r canning  et  to  a l .  and  changes Atlantic  found of  product.  flesh  was  determine  not  (1986)  a l . (1984)  content  canned  under-  maturity  farmed  salmon  raw  et  each  i t was  chemical  female  moisture  from  sexual  needs,  the  content  however  Aksnes  coho  the  l i p i d s ,  examined  of  Bilinski  between  moisture a  male  fish  stage  energy  maturation  final  the  dramatic.  of  total  significantly  lipid,  advanced  became  f o r the  the  errors  fish.  values  protein  by  and  from  lipid  tail  fish.  of the  studa  sig-  the  raw  It  was  potenaccept-  during progressive increase i n  and  analysis done  and tail  the  Thurston  portions of  the  similar  lowest  the  analysis.  lipid centre  -56-  from  and  the  dorsal  reduce  exper-  different  silver  protein  lowest to  from  (1964) t o o k  proximate  the  only  i n order to  a n a l y z i n g muscle  contained was  proximate T h i s was  Karrick  centre,  highest The  at  and  for  differ-  maturity.  muscle  nape,  was  salmon  Samples  the  that  protein  correlation  concluded tial  suggest  significant  scores  4 differed  despite different  samples  the  mean  of  proximate  salmon  revealed a  loss  changes  that  1 and  results  fish  the  content  between  grades  gradual the  found  lipid  parts  samples salmon The  to  section.  but  the  compare  nape  moisture  values,  from  of  had  the  content.  the  amount  3. TEXTURE ANALYSIS  3.1.  Introduction  The  textural  flection tural  of  elements  ment  in  the  constitute the  the  up  rheological  of  to  with  a  tent  than  soft  of  due  of  to  fish,  the  to  the  Hatae a  a l .  Once  found  heat  the  between stated  during  heat  denaturation  of  struc-  proteins  content  in  structure  Unlike in  and  mammalian  relatively  the  (1990)  significantly  fish.  flesh  of  re-  arrange-  g e l a t i n i z e d by  texture  a l . , (1984)  fish  meat.  a  the  fibrous  the  present  et  of  protein  for  easily  are  complex  The  total  fish are  are  had  et  the  of  tissues  textured  Iso  their  responsible  r e l a t i o n s h i p was  texture  rily  are  texture  of  the  a l . , 1974).  tenderness. the  in  and  to  80%  muscle  concentrations  1979).  contribute  firm  no  tissue  properties  not et  ever,  70  and  fish  (Dunajski,  collagenous  do  (Love  the  and  concentrations and  properties  muscle  muscle,  tissue,  c h a r a c t e r i s t i c s of  higher fish  collagen  raw  fish  cooked  processing  conhow-  content  any  fibrous  product  collagen  was  that  cooking,  cooked found  low  changes were  protein  and in  primain  the  tissue. Love of  a l .  steam-cooked  eters of  et  such  the  tween  as  fish. pH  and  (1974)  cod  pH, A  investigated  muscle body  length,  negative  texture.  in relation  As  and  differences to  -57-  pH  number  physiological  c o r r e l a t i o n was the  a  of  the  in of  texture param-  condition  demonstrated muscle  be-  increased,  the  flesh  became more t e n d e r .  correlate not of  negatively with  true  for  spawning  fish  important,  levels  thus  pH  the  Huynh  ing  chum  6.2  i n the  7.0  f o r the  salmon  et  a l .  of  the  as  (1990)  fibres,  of  fish.  over  each  fibres.  In  firm  shifting  of  the  being as  i n pH  f o r the  light  dry  and  formed, would  in  mortem  dark  the  soft,  that  of  be  spawnpH  fish  from  and  pH  the  other  and fish, than  as  fibres  cooked  by  with  texture  was  in  muscle  measured  correlated  after  fibres  separated there in  the of  being  were  appeared  mastication, discrimination  -58-  muscle  scanning  a  thincorre-  fibres.  samples  bundles  and  sections of  friable  muscle  the  bundles  role  firmness  muscle  textured  microscopy  t e x t u r e was  a  longer  the  Cross  with  revealed  Instead,  used  observe  firm,  thicker,  deformed.  to  cooked  A  6.5  most  post-mortem  much  change  was  fish.  compared  Texturometer  During  measured  drop  pH  were  shifted  not  to  stress  f a t , and  acid  to  Observation a  lactic  found  The  Lower  silver-brights  microscopy  with  muscle.  less  did  protein,  increase i n post  shorter  lated  in  (1989) and  cod  cycle.  an  Texturometer. ner,  the  also  relationship  noted  texture  f i b r e s  in  this  spawning  depletion of  muscle  spawned  Hatae  the  the  l e n g t h was  however  their  resulted  of  expected.  electron  the  glycogen  glycogen  pH,  during  caused  Body  was  tested  not  to  have  into  broken  or  slid,  or  individual  considerably  soft  textured  t e x t u r e was  on  less fish.  thought  to  be  achieved  of  muscle  tial ing  et  a l .  and  muscle  absence  of  heat  firmness  and  The  over  Instron  of  This  able  used. has  tural  many  used  force-time are  able  to  This  by  twice  movement  ways  curve or  to  (TPA)  a  calculated  number to  and  determined  the  i s  an  the  imitative  deformation  of  on  i s  a  bite  or  inter-  precision.  mechanical  method  slid-  tissue.  degree  failure  the  a  wide  size  cell  technique  that  variety  the  tex-  mouth's  with  Based  textural  describe  of  the  samples  motion.  of  of  test  simulates  reciprocating  obtained,  the  presence  the  measure  measure  testing  a  in  depending  analysis  the  cooking intersti-  impede  Machine  the  the  into  fibres  high  measure  compressing in  to  muscle  to a  during  Thus  muscle  Testing  successfully  derived,  other.  with  profile  motion  plates,  i s  food  acted  cooked  different  attributes.  chewing flat  i s  Texture  been  sliding  squeezed  protein  the  of  that  characteristics  in  of  Universal  instrument  foods  each  coagulated  spaces  that  were  coagulated,  friability  machine  suggested  proteins  once  fibres  intra-cellular  type  (1990)  sarcoplasmic  spaces, of  d e t e c t i n g d i f f e r e n c e s i n the  fibres.  Hatae process,  by  on  two the  parameters  texture  of  the  food. Aksnes properties changes the  et of  a l . (1986) maturing  i n odour,  immature,  and  studied the  changes  farmed  Atlantic  f l a v o u r , and  texture  mature  fish.  -59-  The  i n the  salmon. were  texture  Significant  observed of  sensory  the  between immature  fish  was  while  described  the  mature  as  filamentous,  fish  became  with  watery,  a  firm  tough  consistency,  and  "distinctly  soft". The tion  biochemical  during  affects  the  the  changes,  spawning  texture  fish  becomes  able  p r o p e r t i e s , and  ration  in  and  of  sensory  canned versal  this  object  The  of  this to  fish  used  fall  of  B.C.),  and  from  the  fish  were  for  several  overnight  inch sions  were thick 307  and  X  were packed  with  a  were  B.C.  washed, After  washed  steaks.  Three  (Wells 213  with  g  deterioac-  commodity. instrumental  changes the  in  Instron  semi-trained  work  from  and  with  made  desir-  consumer  an  textural  the  taste  the Uni-  panel.  Methods  removed,  200  on  develop  were  As  This  Columbian  obtained  Packers  Chilliwack hatchery  graded,  4°C,  the  adversely  i t s firm,  effect  to  starva-  flesh.  watery.  British  measure  salmon  loses  and  was  chum  cooked  muscle  work  1989  months.  at  and  soft  for this  and  of  significant  Measurements  summer  bones  a  M a t e r i a l s and  The  the  becomes  T e s t i n g Machine,  3.2.  raw  important  method  fish.  the  has  physical s t r e s s of  migration  mature,  texture  ceptability The  more  of  and  dressed,  storage,  in cold  the  Can  piece  of the  the  were  the  and  1%  -30°C thawed  and  butchered of  B.C.).  at  were  skin  cans  the  (Richmond,  frozen  fish  Limited,  salmon,  -60-  and  tinplate  Company  Ltd.  (Chilliwack,  water,  fillets  during  major  into  1.5  the  dimen-  Burnaby,  B.C.)  un-iodized  salt  was  added.  for  an  room  ined  This  from  a  an  Instron  the  panel  of  forks,  free  into  was  mental  evaluation  units  and  used  fish  perimental  units,  48  were  cans,  cans,  and  (TPA)  method  severing a  Testing  the  of end  notched  of  and  5  fish;  21  and  cans,  1  8 fish;  fish;  and  8  3,  4 was  was  Machine  by  Bourne of  nylon  a  (1978). 50  disk  cc  Texture Samples  syringe  in  the  -61-  of  for  each  instru-  experimental  2,  4  of  11  ex-  experimen-  experimental  r e p r e s e n t e d by  of  performed the  flaked  7  fish.  Instrumental analysis analysis  of  were  represent  Half  16  1122,  then  half  grade  exam-  fish  to  consisted  grade  grade  each  mm.  and  were  objectively  was  numbers  grade  with  month  (Model  from  3-5  evaluation  and  tested  salmon  The  W  commercial  salmon  combined  about  follows:  cans,  of  cans  canned  500  f o r one  Machine  and  texture.  7  units,  Instrumental Universal  liquid,  FMC CA),  known held  248°F  further.  were  Testing  The  as  33  3.2.1.  were  at  i n an Clara,  on  grades  also  pieces  of  12  experimental  Santa  based  Three  f o r sensory  units,  units,  and  steam  minutes  cans  four  MA.).  experimental unit. two  The  Universal  sample  ing  p r o c e s s was  Canton,  drained  65  Corporation,  of  sensory Instron  of  i n pure  before being tested  each  Corp.,  opened,  using  retorted  1989).  temperature  by  using  tal  were  (FMC  (Collins,  Cans  one  retort  cooling.  practices at  cans  operator's process time  Laboratory water  The  at  texture  using  the  Profile were  of  Analysis  prepared  i t s zero  bottom  Instron  the  line,  by  plac-  cylinder,  adding with of  10  g  of  another  2.5  cm.  geometry  nylon This  and  cm  the  bite-size  were  form  100  cylinders  The  recording intervals  chewiness, Bourne  and  data  over  sociated  and  2  cm  plate to  used  of  Five  measures  was  0.02  interfaced  seconds. were  a of  to  a  of  were  0.06  calculated of  seconds.  compression  was  as  compress  cross-head each  computer  to  digitized  slopes the  to  of  by the  digitized  maximum  equated  sample  described  from  The  A  cohesiveness,  the  calculated  sample  original  in  Hardness,  values  to  their at  i t  diameter  compression.  was  25%  replicate  a  cylindrical  motion  curves  each  of  uniaxial  fish  Incremental  intervals with  compressing  strict,  coated of  lightly  force/deformation curves  of  force/deformation  a  correct  gumminess  (1978).  height  Instron  of  and  reciprocating  mm/min.  collected.  at  for  a  a  ensured  teflon  in  fish  to  system  twice,  of  allow  disk  flat-faced,  height,  flaked  allowed  5.5  speed  the  slope  as-  instrumental  firmness. 3.2.2. Texture General  on  prior  Three  analysis  Foods  Szczesniak  Sensory  Texture  et  panel  to  familiarize  be  expected, and  salmon  was  Profile  method  (Brandt  Nine  experience training  the  and  with by  to  the  et  were  after a l . ,  selected  i n  the  range  the  a  of list  canned  the 1963; based  project.  conducted  consensus  describe  -62-  modeled  interest  s e s s i o n s were  develop  definitions  panelists  and  panelists  to  of texture  the  a l . 1963).  preliminary  attributes  of  analysis  in  order  textures of  to  texture  fish.  The  definitions compress  chosen  included  the material  between  sation  of a progressive  cavity  during  sample); during to  mastication;  prepare  panelists vertical was  on both  scriptions. of  the four  f o r each  anchored  This  ends  3.1  number  ready  the sen-  i n the oral  wetness  of the  of fibres  the total  evident  effort  required  f o r swallowing.  on a 6 i n c h  with  represented  i s an  linear  reference  attribute fora  Figure  fluids  to  The  t e x t u r a l a t t r i b u t e s by marking  sample  mark  the individual  samples.  to a state  required  dryness,  (i.e. the overall  and chewiness,  assessed line  i n free  the perceived  t h e sample  the force  the molars;  increase  mastication  fibrousness,  firmness,  scale  samples  the panelist's sample,  example  of  a  which  and dejudgement  relative  to  the rating  other sheets  used. Ten of  half  the fish  hour  and i n each  unknown  fish  compare  the texture  samples  samples,  flaked  sented  product  a  present,  required each  tuna  a  was  a n d ham.  t o assess  session  each  judge  t o analyze.  a  soft,  effort product  product  made  held  and canned  with  fish  were  juicy  which  were  helped  flesh,  was  firm,  effort.  t h e same reduce  -63-  t o two  very  four  asked  to  reference ham  repre-  few  fibres  The canned  tuna  d r y , f i b r o u s and  Because  over  given  were  The f l a k e d  t o chew.  needed  t o purchase This  tuna.  needed  chewing  samples  the texture  was  Panelists  o f t h e salmon  considerable  reference  effort  ham  and l i t t l e  represented  sessions  many  cans o f  the ten sessions, l o t numbers  variation  an  f o rt h e  within  the  Name:  Date: EVALUATION  OF T E X T U R E A T T R I B U T E S  Please, evaluate the firmness, juiciness, chewiness and f i b r o u s n e s s o f t h e s e samples by marking a v e r t i c a l line for each sample on t h e h o r i z o n t a l l i n e s t o i n d i c a t e your ratings. L a b e l e a c h v e r t i c a l l i n e w i t h t h e code number o f the sample i t r e p r e s e n t s .  FIRMNESS soft  firm  DRYNESS wet  dry  FIBROUSNESS few  many  CHEWINESS little  effort  much  effort  COMMENTS:  Figure  3.1.  Example o f t y p i c a l t e x t u r e s c o r e s h e e t used t o analyze texture o f t h e 4 grades o f t h e canned chum s a m p l e s .  -64-  reference of  the  points  test  samples.  d r i n k f o r one performed visual  which would The  hour p r i o r  s o d a w a t e r , and  "halo  there  effect"  w o u l d be  booths under r e d  tasting  was  of  each  effect,  smoke  was  t o mask  Water,  or  any  unsalted  sample. were  considered  t h a t b i a s would o c c u r  1977)  i n which  the  a general  f o r each  of  the  four  s a m p l e s were a c t u a l l y t h e  ples  were  presented  to  the  the  from  the  To  with  attributes  by  impression  samples.  j u d g e s were p r e s e n t e d  t o l d when t h e fish  to  lights  samples.  texture  concern  (Larmond,  samples  of  not  Sensory a n a l y s i s  formed about the t e x t u r e of the  nate t h i s coded  panel.  between t h e  attributes  j u d g e s and  p a n e l i s t s judgement  u n s a l t e d c r a c k e r s were a v a i l a b l e f o r c l e a n s -  i n g t h e mouth a f t e r Four  the  j u d g e s were a s k e d to the  in individual  non-uniformity  affect  elimi-  differently and  were  same f i s h .  not  The  panelists in a  sam-  random  order. 3.3.  R e s u l t s and 3.3.1.  Figure  3.2  tained  from the  sis  of  the  are  given  values their  of  the  of  a  Instron  typical f o r the  produced  Table  standard  analysis  Instrumental  shows  curves in  Discussion  3.1.  variance  the  data  table  f o r each  of  Statistical  (ANOVA) was  p o t h e s i s t h a t t h e r e was  no  profile  curve  c a n n e d chum s a m p l e .  This  parameters errors.  by  texture  collection  represents the  four  to  Analyprogram  the  mean  grades,  a n a l y s i s by  performed  ob-  a  test  one-way the  d i f f e r e n c e between g r a d e s f o r  -65-  and  hythe  ' I I I !  • M i l l ' M i l l M ; i | i M  i  ;  :  * : ' i i i • , i M I :  •  *  •  I  i  I  i  Hardness 1  Direction of chart  Figure  3.2.  Example o f a t y p i c a l I n s t r o n t e x t u r e a n a l y s i s c u r v e f o r a chum s a m p l e .  -66-  profile  Table  3.1.  Parameter  Mean i n s t r u m e n t a l TPA g r a d e s o f chum s a l m o n .  Grade  1  Grade  Grade  the  Grade  4  four  1  Area  2  Hard  1  8. 3 3 ( 1 .02)  5. 2 3 ( 1 . 2 0 )  6 . 7 4 ( 0 .85)  5 . 0 9 ( 1 . 13)  Hard  2  5. 5 5 ( 0 .72)  3. 4 8 ( 0 . 8 5 )  4 . 5 4 ( 0 .59)  3 . 4 9 ( 0 . 57)  Cohes.  1. 0 6 ( 0 .04)  1. 0 8 ( 0 . 0 3 )  1 . 0 4 ( 0 .02)  1 .16(0. 0 3 )  Spring.  1. 8 3 ( 0 .13)  1. 9 9 ( 0 . 0 7 )  2 . 0 0 ( 0 .11)  1 . 6 4 ( 0 . 18)  (25 • 0)  78. 5  ( 9 .58)  ( 3 2 . 2)  5 6 . 6 ( 1 1 . 5)  170  3  of  Area  202  152  2  parameters  ( 2 0 .0)  70 . 9 ( 1 5 .7)  139  ( 2 9 . 3)  48 . 7 ( 1 1 . 0)  Slope  1  36. 3  ( 5 .89)  1 9 . 2 ( 4 .8 2 )  28 .8 ( 4 .41)  20 .4 ( 4 . 7 4 )  Slope  2  3 8 . 2 ( 6 .03)  2 0 . 6 ( 5 .7 9 )  30 .2 ( 4 .92)  20 .5 ( 6 . 1 3 )  Gummi  21. 5  14. 1 ( 3 .39)  17 .9  14 .6 ( 3 . 1 3 )  Chewi  9 6 . 4 ( 1 4 .1)  7 0 . 9 ( 1 7 . 6)  84 . 1 ( 1 2 .3)  n  3  12  ( 2 .74)  06  14  ( 2 .38)  60 . 6 ( 1 6 . 0) 05  Standard errors are presented i n brackets. Two o u t l i e r s w e r e r e m o v e d f r o m t h e d a t a s e t . "n" = number o f e x p e r i m e n t a l u n i t s .  -67-  2  texture  parameters.  systematic was  increase  discovered.  reflection the  of  texture  square order were  transformed  data  <  were  but  Area  1 1  chewiness  were  Multiple  comparisons  degree  improved  of  grades  1  and  biological  lead  to  2,  high  contributed  to  1,  2,  3  and  (a  for  slope  s c a t t e r i n the standard the  lack of  the  their  Slope  the  much  2  of and  s i g n i f i -  grades  (a  significant  gumminess,  four  were  fish  were  were  1 were  grades  contrasts,  different 2,  slope  of  in  analysis  3.2.  which  one  between  grade  and  Table  springiness,  between  freedom  in  the  in  data  because  of  a  A  Two  measures  results  and  different  cohesiveness,  significantly hardness  of  outliers  least  sequential rejective  parameter between  not  2,  and  as  was  fish.  the  variances.  parameters  at  and  the  replicate  shown  on  a  mean  inherent  spawning  of  The  ANOVA,  variances  performed  set  only  of  in  of  increasing  variation  than  was  fish.  between  hardness  0.10.  single  fish,  between  are  with  homogeneity  data  the  different  errors  homogeneity  the  assumptions  biological  mature  the  from  cohesiveness  a  less  other  0.05),  of  larger  the  cantly  standard  lack  deviations  than  testing  transformation  deleted  higher the  the  improve  standard  in  The  of  root to  Upon  chum  < at  and  grades.  performed  with  and  corrected  with  the  Bonferroni  procedure.  For  the  4  was  <  0.05).  slope 1  (a  <  significantly  2  Grades (a  0.10).  instrumental  deviations significance  -68-  <  of  1  different and  0.05), The  4  were  and  for  high  measurements the  f o r TPA  mean  degree which values  variables.  Table  TPA  3.2  Probability values produced f r o m o n e - w a y ANOVAs on t h e s q u a r e r o o t t r a n s f o r m e d I n s t r o n TPA d a t a .  Parameter  Probability  Value  Area  1  0.28  (NS)  Area  2  0.10  (NS)  Hardness  1  0.06  Hardness  2  0.08 •''O.OOl  Cohesiveness Springiness  0.26  Slope  1  0.06  Slope  2  0.04  2 2 3 (NS) 2 3  Gumminess  0.15  (NS)  Chewiness  0.24  (NS)  Two o u t l i e r s r e m o v e d f r o m t h e d a t a s e t S i g n i f i c a n t a t t h e 10% l e v e l S i g n i f i c a n t a t t h e 5% l e v e l  -69-  Observation quired both  the  on  quired was  most  the  crease  can  changes  in  a  loss  tissue  tent  the  of  the  from  t h e  was  non-collagenous, fish,  the  increased sticky,  moisture  more  content,  cohesive  Reproducibility a f f e c t e d  by  Johnston, texture the  fish. the  (1981)  was  flesh  the  final of  found  middle  et  one  was  there  was  moisture  concon-  an  increase  farmed  Atlantic  of  protein  the  e x p e n s e  of  Upon  retorting  the  gelatinized,  of  of  and  with  formation  the of  a  -70-  was  units.  o b j e c t i v e evaluation of variations  reduced the  measurements  experimental  structure along  a l . (1986) of  of  product.  because  t h i r d  inThis  increase i n  found  objective texture  and  to  effects  mature,  i t contributed to  that the  composition  a t  re-  physiological  metabolizing  proteins.  p r e p a r a t i o n  difficult  Aksnes  the  4  four.  the  maturing  p l a c e  tissue  and  relative  of  found  grade  a l . (1986)  that  cellular  connective  observation  was  more  sample,  this  increase i n the  et  took  and  became  content  concluded  m u s c l e  spawning,  an  Aksnes  to  re-  grade  c o n s i d e r i n g the  fish  proteins,and  muscle.  It  by  to  the  1  while  which  one  proteins, a  hydroxyproline  salmon.  grade  grade  compress  exception  during  the  that  compression,  An  from  in myofibrillar  nective  in  As  to  cohesiveness,  salmon  processing.  showed  energy  explained  the  trends  second  parameter  be  or  force.  significantly  finding  heat  and  least  TPA  general  force,  first,  the  the  of  which the  variation  f i l l e t s  for  existed  length by  fish  of  using  in the  only  organoleptic  analysis.  In  randomly  from  combined  to  greatly on  the  different  obtain  reduced  canned.  were  In  order  to  texture  within  the  tion was  of  f o r the  decision  whole the  chunk,  state  and  in a  fish  sensory lets,  and  but  and  slide  the  canned  which study  the  fillets  of  were  fillets  onto  from  the  and  fish,  the  effectively  cooked  was  correlation  was  based  more  i s  the  can,  a  flaked  was  on  f o r the found  i n the  the  caused  thus  making  -71-  the  served  as  consumed.  instrumental cooked  mince  between  cooked  f i l -  minced  variation  in  minced  the  fish.  of  the  layers  myomere  i t impossible stated  a  simulate  application  S z c z e s n i a k (1987) eliminated  The  observa-  f o r the  higher than  fillets  two  and  of  that  loca-  product  closely  coefficient  found  of  muscle  or  correlation  found  based  evaluations.  a l . (1983),  No  were  technique  in  commonly  fillets  and  locations  commonly  would  salmon  data  force,  analysis.  was  observed.  the  muscle,  sampled  error  variation  sensory  et  This  various  to  i s not  Borderias  e t a l . (1983) test  the  flaking  addition,  cooked  in  the  packing of  salmon  comparisons  In  to  canned by  a  were  introducing  due  samples  instrumental  away  produce  due  flaked  significant  Borderias pression  cans  of  error  instrumental  species  products. raw  reduce  therefore  in  sensory 5  use  First,  Second,  of  to  cans  along  between  connective tissue  used  tions.  possibility texture  that  three  experimental unit.  in  fish  study,  locations  one  the  differences  the  present  that  heterogeneity  a  to  comto re-  mincing of  the  test  material  scatter  in  mincing not  which  the  measuring  tion  the  of  of  the  A  the  high  degree  disadvantage  the  disrupted,  should  fibres  of  of the  could  then  became  a  nearly  The ringe,  orient  cylinders for  within to  found of  inherent  of  strumental  of be  very  of and  texture. sensory  was  total  integrity  of  same  that  the  However  myomeres,  the  became  measure  stress.  the  gelatiniza-  fibres  to  was  ineffective.  recommended  the  the in-  resisbecause  positioning  d i r e c t i o n each  samples  effective  However  could  in  fish,  in  the time  task.  despite unit,  were  differences  fish  It  forming  units  often  muscle  fish's  fibres  incomplete  the  measurement  the  experimental  experimental  grade,  drying  to  fish.  each  the  designed  impossible  technique  was  and  mechanical  of  the  were  encountered  cooked  structural  be  to  of  resulted  resistance.  tests  difficulty  methods  collagen,  of  a  properties  fish  was  the  to  that  most  organization  sample  same  of  elements  tance  tion  test.  stated  instrumental  the  strumental  ples  of  some t e x t u r a l p a r a m e t e r s  rheological  cooking  myocommata  of  that  (1979)  common  Because  only  was  cause  detected.  Dunajski  that  the  instrumental  however  be  was  the  s t i l l  draining also  Because analysis  -72-  5  50  and  caused  can  came  deviations In  fish  from  addiof  the  contents,  or  variations  samples  sy-  identical  high.  between  the  cc  r e p l i c a t e sam-  standard quite  of  have  a  in preparing having  within  in  for the  both  in the  same  the in-  three  cans, ble  instrumental  after  sulted  sensory  i n some  tributed  measurements analysis  drying  to the  high  3.3.2. Table tions  3.3  from  of  the  the  Sensory  sensory the  analysis  variance  was  there  each  attribute.  cause the  was  each  other  no  attribute  judges  ing,  linear  scale  tended  to  judge  to  to  an  only  and  delay  re-  have  con-  may  texture and  standard  evaluation grades  performed  of  to  between  least  of  the  chum  test the  the  four  grade  devia-  textural  salmon.  rejected  one  tended the  An  hypothesis grades  for  however,  be-  different  this  variance  by  different  of  from  the  further  to  the use  of the  between  there  than scale  judges,  was  trainof  the  judge  "X"  judge  "Y"  scale,  and  end  that  attribute,  and  grades  the  the  n o n - u n i f o r m i t y was  out,  parts  scale, of  individ-  their  example,  middle  textural  the  despite  For  i n the  other  between  use  half  results  used  that  samples.  first  non-uniform  non-homogeneous  to  f o r each  each  system  conclusion  the  of  average  the  the  the  effect  difference  scoring  evaluations  cancelled  eliminate  the  mark  marked  The  culating  of  h y p o t h e s i s was  judges  a l l the  scale.  less  use  "Z"  values  of  lead  different  grouped  muscle,  analysis  at  This  possi-  deviations.  four  had  only  three.  Observation ual  fish  difference This  were  completed.  panel's  between  that  texture  t h e mean r e s u l t s  differences of  was  standard  displays  of  of  the  when  cal-  the  appeared  extreme to  expected.  which  resulted  be To in  a Z-transformation  Table  3.3.  Mean, u n t r a n s f o r m e d r e s u l t s f r o m t h e s e n s o r y panel's evaluation of the textural differences b e t w e e n t h e f o u r g r a d e s o f chum salmon.  Grade  Sensory A t t r i b u t e s Dryness Fibrousness  Firmness 1  7.69(2.6)  2  Chewiness  n  8.34  (2.3)  8.60  (2.3)  7.41  (2.8)  97  6.81(3.5)  7.16  (3.2)  7.87  (3.0)  7.09  (3.0)  43  3  6.54(2.8)  5.22  (2.7)  7.00  (2.8)  7.42  (2.7)  150  4  3.15(2.3)  3.76  (2.5)  2.82  (1.6)  3.15  (2.2)  62  1  Standard n= t o t a l grade.  d e v i a t i o n a r e shown i n b r a c k e t s . number o f o b s e r v a t i o n s f o r a p a r t i c u l a r  -74-  was  performed  judge  on  individually  was  equal  was  justified  values  f o r each  showed  a  grade trend  chewiness,  vealed  a  The  the  data.  the  sequentially  the  overall  this  comparison  and  between Grade  dry.  For the attribute  less  which  distinguished,  fibrous  ness,  no were  than  t h e most  attribute  3.4.  of the 4  grade  3,  found  firm.  3  chewiness,  firm  to  4 was 1  control  grade the 2  2,  2,  least could  significantly  3 and 4 could  1,  to  attribute  and  grades  grades  of  able  For the attribute  than  with  results  than  grades  with  hypotheses  were  and 4 were  grades  -75-  tested  f o r the  dryer  between  Grades  less  judges  grades  dry-  1988)  contrasts  The  and grade  and grade  and were  1987).  fibrousness,  was  (Wilkinson,  procedure  The  graph  one o f t h e  f o r multiple  any o t h e r grade.  difference  distinguished, the  than  at least  o f t h e d a t a were  significantly  2  be  dryer  was  grade  not  was  1  each  bar  of variance r e -  o f freedom  rate  samples.  i n firmness,  Systat  Copenhaver,  i n Table  The  Analysis  Bonferroni  error  DiPonzio  are given  distinguish dryness.  rejective  This  of the standardized  between  comparisons  t h e same  decrease  degree  scores  t o one.  attributes.  a  f o r each  of their  equal  tested  program  single  Multiple  test  of the 4  difference  t o perform  o f each  representation  statistical  used  attributes  was  and f i b r o u s n e s s .  was  (Holland  judge  towards  significant  grades.  t h e mean  each  i s a graphical  clear  the textural  and t h e v a r i a n c e  because  3.3  ness,  of  so that  t o zero  Figure  4  each  1  firm-  versus  also  n o t be  1 a n d 2.  and  3  2  were  For not  -2  1  1  GRADE  1  GRADE  2  SEXUAL  Figure  3.3.  GRADE  3  GRADE  4  MATURITY  Standardized sensory scores f o r the four grades o f chum salmon f o r the attributes firmness, chewiness, d r y n e s s and f i b r o u s n e s s .  -76-  Table  3.4.  R e s u l t s from a n a l y s i s of v a r i a n c e of the sensory s c o r e s t o determine d i f f e r e n c e s between g r a d e s , and s e q u e n t i a l r e j e c t i v e Bonferroni procedure f o r the m u l t i p l e comparison of the data.  Effect 1 vs  2  1  Difference within the grades vs 3 1 vs 4 2 vs 3 2 vs 4  3 vs  Firmness  NS  *  *  NS  *  Dryness  *  *  *  *  *  *  Fibrousness  NS  *  *  *  *  *  Chewiness  NS  NS  *  NS  4  *  * = significantly different at a = 0.05. NS = 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 a t a = 0.05.  -77-  significantly 3.3.3.  different,  Sensory  Correlation  and  and  sensory  Systat  (Wilkinson,  between  the  they  was  values 1988).  parameter  using  a  There  was  Table  i s part  instrumental tion  and  the  sensory  use  of  sensory  or  quality  of  a  be  a  of  accuracy  loss  samples  are  examined  will  mean  (1982)  commented  non-linearly they  analysis,  The  a  problems  much work  on  Levitt, whether sory to  an  useful  to  to  4.  the  significant and  the  from a  by  correla-  attributes  juiciness  with  instru-  correlation  (a  =  0.05).  comparing  Pearson  fatigue  of  time,  i f a and  members  o f t e n the  the  the  correla-  physical  and  that  the  responses  were  chemical  data  which  effective  First  the  panels  could  steps be  sensory  used  used  be  variables  changes sensory  quantified processing.  have  prompted  techniques. to  i n place  organoleptic characteristic.  -78-  used  of  instrumental  described three  instrumental test  could  effects  sensory  panel Lund  generated  the  of  team.  s t u d i e s have  predict  may  number  unavoidable  many  to  there  large  of  sensory  acceptability  Although  developing  the  evaluate  disadvantage  short  associated with  (1974)  to  the  loss  that  have  evaluations.  describe  due  the  reflect.  model  has  in a  related  few  panels  product  changes  into  a  created  attributes  grade  matrix. The  that  between  Pearson  and  matrix  than  correlation  cohesiveness  f i b r o u s n e s s , chewiness, of  chewier  performed  firmness, 3.5  were  instrumental  analysis  mental  tion  but  were  determine of  sen-  selected  Instrumental  Table  3.5.  C o r r e l a t i o n v a l u e s ( r ) e x t r a c t e d from a Pearson c o r r e l a t i o n m a t r i x o f t h e comparison between t h e instrumental parameters, and t h e sensory attributes.  Instrumental Parameters  Firmness  Sensory Dryness  Area  1  0.347  0.239  0.324  0.366  Area  2  0.432  0.235  0.404  0.451  Hard  1  0. 387  0.197  0.323  0.375  Hard  2  0.376  0.182  0.316  0.378  Cohesiveness  0.788*  0.521*  0.827*  0.815*  Springiness  0.411  0.325  0.538  0.489  Slope  1  0.388  0.176  0.292  0.379  Slope  2  0.410  0.182  0.328  0.413  Gumminess  0.314  0.143  0.237  0.324  Chewiness  0.344  0.200  0.319  0.385  *  Sample s i z e Significant  'n' = 19. a t t h e 5%  Attributes Fibrous.  level.  -79-  1  Chewiness  variables  were  information step  was  then  contained  to  described  judge  the  the  by  If  sensory  quickly,  Aguilera analysis  to  be  instrumental cluster  and  discriminant  neously.  Other  l i m i t e d because  bers  varied  was  with  tural  within  magnitude  was  applied  Instron's  often force  such  same to  mea-  function  obtain  the  analysis  a  product  as de-  more  as  set  (Lund,  an  applied  compression  a  of  stepwise  how  regression  one  set  parametric and by  simulta-  of  num-  data  i.e.  correlation  correlation  of  1982). load  and  the  (1973)  a  rate  evaluation found  sinuosoidal  t e s t was  -80-  linear  affected  and  simplification  regression  Sherman  in  and  variables  simple  instrumental  Shama a n d  occurred  of  sensory  techniques  efficient  assumed  were  of  The  indicated  and  multivariate  analysis,  number  only  influenced  properties.  for  for  large  results  of  adequately  variables  considered  texture.  Multiple  each  The  chewing  of  another,  l i m i t e d because  load  a  they  distribution.  variables  approach  methods  were  normal  (1990)  allowed  of  the  third  organoleptic  that  possible  p r i n c i p l e component  interpretation  the  c h a r a c t e r i s t i c of  measurements  analysis  The  the  economically.  best  analysis,  of  shown  be  describe  variables  fulfilled  i t would  Stanley,  the  be  could  variables.  dimensions  methods  and  they  instrumental  organoleptic  accurately  that  sensory  i t could  panels,  of  so  the  underlying  instrumental  scription  and  in  whether  characteristic. sured  selected  linear  that of  that  the tex-  while  pattern, application  the of  stress tion  t o t h e sample.  between  instrumental  leading.  However,  the  stroke  down  linear  motion),  brittleness, Hatae fish  by  fibre  individual sensory of  relation  and  might  methods and  differences  sensory data  tests  misduring  approximately hardness,  sides  which  generated  by  using  firmness,  fibre was  nature,  operated  could  analysis.  behind  of  were  between usually  underlying  or  different  Therefore  cor-  o r an un-  the association. different with  These  r e s u l t s were  -81-  evaluation  equality  and minced  instrumental  sensory  analysis.  fish  were  then  compared  compression,  Instron  be  i n t h e same  fillets  an  con-  could  and r e f l e c t e d  studied  shear,  fish  and  using  I t was  correlations  not imply  cohe-  performed  of the correlation.  of  diameter,  measurements  correlated best  skinned  of 5 species  b u t n o t by The  texture  have  TPA methods.  performed  (an  of various  analysis  e t a l . , (1983)  cooked  often  t o measure  volume,  analysis  o f t h e mechanism  to find  correla-  properties  weight,  and d i s c r i m i n a n t  prediction  Borderias  that  was  the texture  o f measurement.  of both  derstanding  the  fibre  instrumental  which  able  residual  an a s s o c i a t i v e , i n d i r e c t  directions  by  drip,  Statistical  items  cycle  was  studied  discriminant  and  effects  Raw  (1984)  textural  by  data  textural  the chewing  of variance  detected  sensory  evaluating  penetration,  that  and  speculated  crispness.  determining  analysis  i t was  the Instron  and  length.  cluded  by  of  et a l .  siveness,  Thus,  and  Texturometer.  tested to  juiciness The  fish  minces  correlated  techniques tal  than  testing  puncture  mackerel ture  and  and to  be  (r=  0.83).  first  bite", Dulce  flaked, panel  value  tached  to  surements 0.91)  with  and  et  (1987)  fish an  juiciness  Ottawa  texturometer  ceived  by  In  a  this  taste  TPA  while all  TPA  parameters sensory  in  were  was  of  more  good  and  of  It  Brabender Brabender  of  machine.  was  predictor  to  de-boned, sensory cell  Force  atmea-  tenderness  concluded of  the  chewing".  extrusion  both  curve  than  semi-trained  with  an  "response  texture  a  tex-  from  Instron  panel's  the  the  the  the  prolonged  with  a  successful  sensory  0.87). a  whiting  obtained and  texturometer  sensory  analysis  discriminating consistently  dryness  four grades.  height  correlated  (r=  the  (r=  that  texture  as  the per-  panel.  study,  instrumental  data  Universal testing  highly  cell,  evaluation of  c e l l ,  compared  Ottawa  blue  beginning  was  evaluated  Instron  were  the  the  the  instrumen-  shear  canned  sensory  peak  instrumental  Effective  Kramer  used  "structure after  with  and  the  and  juiciness.  shear  Instron  a l .  an  the  with  at  predicting  canned  and  whether  The  The  in  (1985)  Kramer  sensory  fillets.  evaluation of  a  correlated  the  included  correlated  with  Brabender  with  fish  Schreiber  Farinograph. curve  the  determine  could  Instron  did  methods  test,  Karl  better  was  was  power.  different  significantly  Correlation  of  -82-  clearly  the  superior  None  of  the  to 10  between  grades,  different  between  instrumental  parameters  and  sensory  with other  attributes  a l lfour  sensory  significant  showed  that  parameters.  correlations.  -83-  cohesiveness However  correlated  there  were  no  4.  4.1. The the  spawning  for of  concluded  its  poor  the  of  ture  analysis  to  in  lipid  from  72%  of  from to  or  80%  understanding  for  (1983)  other  found  were  muscle 1.6%,  early  no  and and  in a  20%  an  evidence  i n the  chum  fish,  Japanese  16%,  increase  late-run  for  (1983)  decrease  to  is  reasons  Konagaya  showed  from  There  responsible  and  and  during  the  parasites  changes  stages  to  salmon  established.  characterize  the  8.5%  chum  a l . (1983),  maturity  content  in  Konagaya  et  of  well  interest  Hatano  different  protein  now  biochemical  texture.  at  quality  sporozoan  that  proximate  crease  i s  deterioration.  presence  waters  eating  migration  and  used  in  considerable  this the  ACTIVITY  Introduction decrease  however,  PROTEINASE  a  in  chum  in de-  mois-  respec-  tively. Mommsen bolic  enzymes,  ble  protein  end  of  and  of  dicating energy  and  or  enzyme.  was  Two  activity, I t was  during the  reduced  to  alanine  thought  of  soluble  and  of  30%  of  of  of  their  was  the  gonads,  showed  aminotransferase  that  these  -84-  original by  enzymes  (GTP), were  By  the  of  the  level.  70%, and  no  metainsolu-  eleven  decreased  enzymes  13  migration.  activity  development metabolic  activity  spawning  insoluble protein in  the  concentration  migration,  i t s use  source.  the  sockeye  enzymes  soluble  level  a l . (1980) measured  spawning  thirteen The  et  in-  as  an  change  in  and  malic  selectively  maintained  in  physiological tivity the  order function  i n muscle  distance  levels  were  change,  or  6.8  8.5.  and  the  t i v i t y  pH  No  low  number  of  cation  of  served  increase of  A,  and  two  so  that  end  pre-migratory were  that  the  indicated  activity  lead  to  activity  of  enzyme m o l e c u l e s ,  D. of  a c i d i c  were  also  migration  was  de-  i t was  concluded  that  the  the  and  ac-  cathepsin  enzymes,  pH  w i t h  involvement  increase.  was  at  during  thus  d i d not  No  proteinase  the  lysosomal  with  s t u d i e s  presence  ac-  levels.  detected  the  or  migration  B-N-acetylglucosaminidase  lysosomes  in  to pH  in  D  increased linearly  i n h i b i t i o n  enzymes,  cathepsin  Proteinase  in activity  Two  change  f o r these  number  the  migration.  concluded  due  lysosomes.  assayed.  total  was  biochemical  4.05  the  on  workers  at  carboxypeptidase  tected  times  decreases  4.0  activity  pH  essential  migrated  Based these  muscle  throughout  salmon  seven  pepstatin,  The  sustain  buffered to  slight  at  to  conclusion  due not  Partial  to  a  an  that  purifithe  increase  result  of  ob-  in  enzyme  the  acti-  vation. Ando  and  Hatano  N-methylhistidine myofibrillar tained  only  protein into amino  (MeHis)  degradation in actin  synthesis or  the acid  urine. in  the  (1986b)  It  and  i n  muscle  an  vivo.  myosin,  energy, was  as  used  and  index This  i s not  extracts  that of  amino  of  acid  utilized  the  a c i d  maturity  amino  i s excreted  suggested  -85-  the  i s  for  of con-  either  quantitatively level  maturing  chum  of  this  salmon  reflected This  the  approach  degradation also  found  than  for  clearly  that  the  a l .  androgens  and  degradation  tivity  level  of  and  in  to  high  protein degree  spawning MeHis  was  that  were  was  degradation.  of  myofibrillar  migration.  It  higher  females  of  spawning  initiated to  nutrient  i n order  to  serum  sex  closely  levels  during  response  levels,  found  estrogens  catabolism  changed  that the  during  (1986)  salmon muscle  tein  myofibrillar  showed  increased  et  chum  of  for  was  males.  Ando as  degree  steroids  related  proteinase migration.  by  hormone  the  protein  activity Enhanced  levels  d e p l e t i o n and  provide  to  such  energy  of pro-  which  increased  ac-  necessary  for  migration. All total of  the  protein,  consider the  The  3.5,  the  the  that  myosin, during  changes  of  activity  6.2  may  the of  and  the  be  apparent  by  migration. i t was  physical  in loss  proteins, i n -  degraded  chum,  the  decrease  the  myofibrillar  spawning  in  as  muscle In  order  necessary  to  degradation  of  body.  object  lysosomal of  and  such  i n MeHis,  p o s s i b l e mechanisms  fish's  the  increase  activated  understand  evidence,  suggested  a c t i n  proteinases  in  the  myofilaments,  cluding  to  available  and  this  level  7.0  enzymes  study of  during have  myofibrillar  an  was  muscle  to  proteinases  spawning important  component  of  -86-  determine  the  the  buffered  migration. role  i n the  total  changes to  pH  Because degradation  body's  protein,  the  activity  also  acid  Materials  The  decision  made  planned which  when  was  were  placed  on  to  side  of  fish  samples and  dark  fish  vessel,  this  time  muscle stored  was at  fish  on  the  The  was  samples had  (Port  year,  test  obtained  sealed  in  1989  days  emphasis  and  a  of was  to  the  taking  the  Department  a n o t h e r DFO  River  3-5  vessel.  River.  in  travelling  aboard  on  Fraser  Chilliwack  by  fishery  work  state.  McNeil),  Strait  the  collected  collected  Island  1990  of  undergone  second  year  Intermeditest  Spawning  Hatchery.  ziplock  of  fishery  fish  Fish  freezer  were  dorsal  bags  and  -30°C.  proteinase  method  of  The  parts  distilled  dorsal  Ultra-Turrax added were  activity  Konagaya  4.1.  tubes  some  were  the  collected,  that  i n a pre-rigor  4.2.1. Measurement  was  marker,  i n the  fish  (DFO)  were  the  the  Johnstone  and  an  lysosomal  samples  which  In  the  Oceans  from  on  fish  Vancouver i n  chum  discovered  changes.  obtaining  Fisheries  collect  post-rigor  N.E.  the  a  Methods  possible  Silver-bright  obtained  and  i t was  not  post-mortem  ate  phosphatase,  assayed. 4.2.  was  of  muscle  of  the  (1983),  and  mixture  homogenizer. of  2 sets  then diluted  muscle  with  Two  of  -87-  was  measured  at  0°C  grams  of  0.2  in  homogenized f o r 30 of  3 triplicate  2 mL  activity  i s described  o f t h e c h u m was  water/ice  t o each  of proteinase  M  the test  by  Figure  in  three  seconds  in  homogenate tubes.  The  citrate-phosphate  Muscle + ice, 1:3  w/w  homogenize at 0 ° C , 30s 2 g homogenate 2 mL citrate-phoshate buffer, pH 3.5, 6.2, or 7.0. (6 aliquaots / pH)  Test incubate  Blank  1 h at 3 7 ° C  Add cold T C A stock  1 Add cold T C A solution to  i  to  15%  Hold at 4 ° C . 1 h.  15%  I Clarify by filtration  I  Hold at 4 ° C , 1 h.  Read absorbance at 2 8 0 nm Clarify by filtration  I Read absorbance at 2 8 0 nm Figure  4.1.  Flow diagram o f t h e method used t o measure t h e proteinase activity i n the d o r s a l muscle of f o u r g r a d e s o f chum m u s c l e .  -88-  buffer  adjusted  to either  three  t u b e s were d i l u t e d  tubes  w i t h p H 6.2  One  the  incubated  bath  and  added 1  shaker  paper.  280  nm  on  a  of tubes  bated.  bath,  were  subtracted absorbance  of cold  vortex  and then  values  filtered  mixed,  filtered  160  U.V  from  value f o r each  Visible Japan).  the  water  ( T C A ) was  was  #2  measured  samples  #2  measured Recording second  not incu-  i n an i c e  f i l t e r  paper.  a t 2 8 0 nm.  The  of the 'test'  were  values  of the four  Whatman  immediately.  f o r one hour  Whatman  'blank'  hour,  The  pH v a l u e s w e r e  held  the t r i p l i c a t e t h e mean  -  Kyoto,  through  1  was  1 5 % T C A was a d d e d  of the f i l t r a t e  from  through  of the f i l t r a t e  Corp.  7.0.  T h e m i x t u r e was h e l d f o r  (blanks) a t the three  absorbance  mean  (Shimadzu  I n s t e a d , 8 mL  tubes  The  acid  U.V.  of three  After  15% t r i c h l o r o a c e t i c  Shimadzu  of  i n c u b a t e d a t 37 °C i n a  from  before being  sets  2 sets  removed  The absorbance  Spectrophotometer  The  were  Two  o f 3 t u b e s w i t h pH  pH w e r e  samples  of cold  i n the cold  filter  set  3.5 b u f f e r ,  and 2 s e t s  a t each  o r 7.0.  a t a p p r o x i m a t e l y 1 0 0 RPM.  (test)  mL  6.2,  t o t h e i n c u b a t e d homogenate.  hour  at  8  3.5,  w i t h pH  buffer,  s e t of the tubes  water-bath  pH  t o obtain  grades  a  delta  a t the three  pH  values. 4.2.2. M e a s u r e m e n t The sured  using  dorsal 18  g  acid  phosphatase  t h e method  muscle  distilled  of acid  activity  o f Beeken  o f t h e chum  was  phosphatase  o f t h e muscle  and Roessner  homogenized  w a t e r / i c e ) f o r 30 s e c o n d s  -89-  was  mea-  (1972).  The  (0.5 g i n an  activity  tissue  i n  Ultra-Turrax  homogenizer, paper. the  and then  This  enzyme  reagent HC1,  solution  1,  and reagent  utes  was  2,  formed the  mixture  read  at  3 mL 405  o f 0.1  nm.  p-nitrophenol and  1.0 mM. of  each  absorbance 4.3.  The  added  citrate  was  Standard  curve with  mL  of  The  f o r 30  minper-  removed  from  and t h e tubes  were  then  were 0.2,  was p r e p a r e d 3  4.8.  were  were  solutions  pH  of  i n 0.001N  Assays  added,  The  volumes  a t 37°C  a t 1 0 0 RPM.  was  (PNPP).  equal  buffer  filter  t o 1 mL o f  phosphate  incubated  NaOH  #2  prepared 0.4,  with  0.6,  0.8,  by t r e a t i n g  0.15  NaOH,  and  reading  a t 4 0 5 nm. Results  and D i s c u s s i o n  4.3.1.  Proteinase  activity  activity  o f muscle  proteinases  tions  present  the  typical  and  pH  7.0  A  was  pH  used  alkaline  are given  8.1.  p H o f 3.5 r e p r e s e n t e d  i n the lysosomal  muscle  3.5 v e r s u s  Table  N  s o l u t i o n  pH v a l u e s .  ment  Na  The samples  standard  three  slightly  was  by mixing  (PNP) o f c o n c e n t r a t i o n s A  Whatman  p-nitrophenyl  shaker  i n triplicate.  bath,  ( 0 . 2 mL)  prepared  0.1M  i n a water-bath  through  p - n i t r o p h e n y l phosphate  0.4% d i s o d i u m  substrate-enzyme  pH  solution  substrate solution,  substrate  mL  filtered  measured  to detect  index  4.1.  was  a  o f muscle,  after  resolution  The r e s u l t s The b a r graph  of maturity  i s shown  significant  -90-  measured  the acidic  the activity  proteinases.  i n Table  There  system  was  pH  at  condi6.2  of  was  rigor,  of neutral,  or  of the experiof activity  at  i n t h e Appendix i n  correlation  (a <  0.05)  Table  4.1.  Proteinase a c t i v i t y a t pH 3.5, during t h e d i f f e r e n t stages migration.  Grade pH  3.5  Proteinase Activity p H 6.2  6.2 a n d 7.0 o f spawning  p H 7.0  1  0. 0226 ( 0 . 018)  -0 .0163 (0 .017)  0 .0219 (0 .014)  1  0. 0369 ( 0 . 024)  -0 .0041 (0 .006)  0 .0344 (0 .006)  1  - 0 . 0129 (0. 010)  -0 .0310 (0 .034)  0 .0256 (0 .005)  2  0. 1453 (0. 025)  0 .0799 (0 .005)  0 .0468 ( 0 .080)  3  0. 0792 (0. 049)  0 .0347 (0 .049)  0 .0180 ( 0 .011)  3  0. 2365 ( 0 . 028)  -0 .0856 ( 0 .002)  0 .0976 ( 0 .001)  4  0. 1295 (0. 013)  -0 .0015 ( 0 .012)  0 .0719 (0 .013)  4  0. 1060 (o. 008)  0 .0144 (0 .024)  0 .0719 (0 .013)  4  0. 2811 (0. 008)  0 .0714 (0 .014)  0 .0976 (0 .001)  Means o f t r i p l i c a t e deviations a r epresented  determinations, i n brackets.  -91-  standard  between value the is  increased  of  0.701,  proteinase shown  in  proteinase Table rity  8.3,  and  The  findings  dorsal  muscle  tivity  was  a  2,  and  no  at  high  i s  shown  r of  maturity  significant  factors.  in  between  an  graph  of  no  two  the  The  Appendix,  activity  activity  migratory  stages.  pH  3.5  at  in  activity  was  kaline  region,  in  the  each  stage  and  matu-  presence  pH  The 3  1 to  stage  3.  to  (1982,  the  1- mM  of  stages. 1  in  acid  ac-  There  to  stage  Because  slightly  calcium,  a l -  and  proteinases the  chum  autolytic  stage  neutral  the  activity  from  of  contribute  in  Konagaya  the  detected  activated to  by  of  activity  proteinase  considered  these  with  bar  index  however  optimum  little  not  8.2,  the  from  were  versus  study  increase  calcium  The  the  increase  cysteine,  9.  proteinase  with  three  even  maturity,  pH.  fold  mM  six  6.2  7.0  compared  at  of  correlation  of  worker  fold  pH  this  highest  four a  at  at  consistent  This  pH  between  activity  1983).  at  size  found  was  found  was  sample  increased  Table  however  are  and  Appendix,  was  range  a  activity  the  correlation  activity  2  (CANP)  changes  in  the  i s  low  muscle. In  mammalian  systems,  skeletal  lysosomal  content.  However,  Reddi  that  catheptic  activity  of  the  greater  than  intracellular skeletal  that  of  muscle  of  of  flounder,  -92-  et  fish  mammalian  localization  muscle a l .  (1972)  muscle  tissue.  catheptic Reddi  et  very  reported  was  ten  To  study  activity a l .  in  times  in  (1972)  the the used  centrifugation fractions. and  These  nuclei,  although At  pH  less  and  was and  t h e P3  found P4  a n d a t pH  enhanced  muscle as  a  to  cells.  Acid  lysosomal  (Chayen  e t a l . 1973).  the different  sulted ganic the  this  phosphate.  colourimetric structed  as  Addition  analysis.  phosphatase  a graph  6.5  could  there  be  detected.  activity a t pH  found  3.5  i n and  was  of  be the  recognized  levels of  i t sa c t i v i t y  was  o f t h e enzyme  NaOH  of the  lysosomes monitored  with  a t 405  i n Figure  reported Table  was  -93-  and  inor-  curve nm 4.2.  measured of  PNP  into by  was  con-  t o phosphate  con-  The  i n t h e Appendix, 8.4.  PNPP r e -  c o n v e r t e d t h e PNP  standard  absorbance  i n Appendix,  might  around  commonly  increasing  i o n , which A  i s shown  was  levels.  incubation  phenoxide  and  a t pH  o f t h e b r e a k d o w n p r o d u c t s PNP  to convert  centration, acid  assay,  activity.  some  concentration  Therefore,  maturity  had  also  has been  that  highest fraction,  of proteinases  such  The  lysosomal  no a c t i v i t y  increasing  i n formation  y e l l o w  (P4),  phosphatase  marker  indicated  In  (P3), lysosomes,  optimum,  phosphatase  five cells  lysosomal cathepsins  enzyme  at  7.5,  into  ( P I ) , unbroken  i n t h e P3,  was  activity  protein  supernatant.  fractions  activity  4.3.2. A c i d  attributed  included  (S4), final  t h e P2,  activity,  The  the muscle  fractions  activity  4.0  separate  (P2), mitochondria,  microsomes, catheptic  to  activity  T a b l e 4.2  of and  1.00  0.80  >  0.60  D  0.40  0.20  0.00 0.0  0.2  0.4  0.6  Concentration of F i g u r e 4.2  0.8  1.0  PNP  S t a n d a r d c u r v e o f t h e a b s o r b a n c e a t 405 nm v e r s u s t h e c o n c e n t r a t i o n o f £-nitrophenol.  -94-  Table  4.2.  Grade  Acid phosphatase activity (mM p h o s p h a t e ) i n d o r s a l m u s c l e o f chum s a l m o n d u r i n g spawning migration. Phosphate Release  1  0.2541  (0.000)  1  0.2770  (0.011)  1  0.2826  (0.005)  2  0.2903  (0.003)  2  0.2943  (0.007)  3  0.3116  (0.010)  3  0.3071  (0.016)  4  0.3401  (0.022)  4  0.4060  (0.001)  4  0.4611  (0.003)  (mM)  2  Means o f t r i p l i c a t e determinations. S t a n d a r d d e v i a t i o n s a r e shown i n b r a c k e t s .  -95-  Correlation correlation increased  (a  The  between  grades  acid  <  0.05)  activity  0.829.  activity  analysis  at  pH  that  softening  of  acid of  and  of  was of  with with  salmon  a  with  an  and  r  value  of  observed  be  muscle  increase  responsible  tissue  of  proteinase  increasing maturity, may  the  significantly  levels  the  significant  maturity  increased  increased  cathepsins  chum  level  enzyme  The  activity  the  the  together  lysosomal  there  phosphatase,  the  4.  3.5,  phosphatase  gested  between  activity 1  showed  in  sug-  for  the  during  spawning  proteinase  activity  migration. The in  the  effect  muscle  (1988)  during  during  crease It enzymes the  the  i n the 4  activity  the  of  expected also  the was  extract of frozen  cold  that  short low not  of  the  each the  time  tropical  fish.  dif-  the  were  enzymes  and  cathepsin  however  to  the  rate  general  of  de-  species. of  time.  chum's However  storage,  temperature  -96-  Significant  aminopeptidase  frozen  experiment.  species  a  with  expected  of  fish  and  was  activity  storage  et a l .  There  fish  in  Warrier  aminopeptidase  storage.  storage,  decrease  7  of  activities  species of  the  activity  specific  activity  on  consideration.  in  and  varied with  would  and  muscle 0°C  months  widely was  change  different  relatively  months, in  at  between  i n the  decrease B  i n the  storage  important  the  storage  ferences found  B  cold  i s an  studied  cathepsin  of  of  invalidate  proteolytic because less  -30°C, the  than  the  of 2  loss  results  of  There ing  has been  the natural  Steiner muscle  in  the  (1984)  microscopy salmon. revealed muscle.  a  histochemical acid  non-uniform  activity  myofibrils.  were  examined  tions  of  lysosomes  their  location  was  cells  by  could  within  at  the periphery muscle  concluded  that  but  not uniformly  were  there  was  lysosomes  discrete  electron of  coho  sections i n the  appeared  t o be  o f t h e endomysium located  and  among  the  lysosomes  at the peripheral  cell  membrane  Epon  The  Three  muscle  among  occurred  as  The  o f lysosomes  popula-  were  of  found  macrophages),  cells,  i n salmon  tis-  the basis  and  the myofibrils.  distributed.  -97-  on  lysosomes  (identified  localization  embedded  microscopy.  distinguished  cells  distributed  and  e ta l .  whether  of, but within,  cells  obtain  lysosomes  lysosomes  centrally  the tissue.  the connective tissue  of  cryostat  of  compartments,  be  of con-  Steiner  activity  area  found  electron  in  the  micron  phosphatase  enzymes i n  i n t h e muscle  distribution  i n the connective tissue  sues  muscle,  phosphatase 10  cells.  In order t o  location  In order t o determine  t h e muscle  i n muscle  techniques f o r light  of  regard-  i n macrophages  and  striated  workers  lysosomal  cells.  to the connective tissue No  that  either  muscle  micrographs  A l l of the acid  were w i t h i n or  normal  to detect  perimysium. the  be l o c a t e d  among  lysosomes  the occurrence  of  Light  of  suggested  or within  about  cells used  limited  might  tissues,  information  disagreement  occurrence  e t a l . , (1984) tissue  nective  much  muscle  within I t was tissue,  conclusion i n coho  that muscle  provided portant It  further  that  i n the deterioration  would  lysosomes spawning  be i n t e r e s t i n g i n intact migration.  involvement during  evidence  e n z y m e s may  o f chum m u s c l e  during  be  muscle  T h i s would  during provide  proteinases  migration.  -98-  different  stages  more e v i d e n c e  i n muscle  im-  spawning.  t o d e t e r m i n e t h e numbers and s i z e  chum  of lysosomal  spawning  lysosomal  of of  of the  degradation  5. STRUCTURAL CHANGES IN MUSCLE  5.1.  Introduction  Electron ture  of  ders  can  lished  both  normal  be  studied.  body  structure niques  microscopy i s a technology  of  i n  Stanley luctance  on  t e x t u r e  have  ical,  of  to  borrow and  has  electron  processing  and  have  been  m i c r o s c o p y may  during There  spawning are  many  ultrastructural physical  be  appearance  stresses  of  useful  tech-  changes  which  a  In  1976,  certain  re-  investigating  meat  microscopy.  Instead,  much  of  the  empirical  as  removing in thin  of the  of  the  flesh  methods. skin  and  the  combine  Light,  fish's  to  of  chum  brine  retortable and  elec-  i n muscle  ultrastructural  migration,  -99-  mechan-  bone,  profile  1989).  the  which  re-  Different  t o observe changes  at  of  study  as  muscle.  texture  employed  factors  biological  level. was  estab-  such  (Collins,  migration  well  sources  processing  tested  disor-  from  a d d r e s s e d by such  muscle  of  study  there  t o t h e complex  improve  heat  to  struc-  the  range  scientists  properties  techniques  treatments, pouches  to  been  food  that  techniques  directed  desire  wide  ultrastructural  the  or rheological  salmon  The  the  part  a  the  i s a  regarding  established  commented  t i m e was  The  tron  at  and  which  striated there  available  tissue been  result,  Swatland  histochemistry search  a  knowledge  muscle  and  pathological  As  of muscle  which  occur  and  by  level.  a f f e c t  skeletal  together  the  muscle.  with  the  process  of  sexual  myofibrillar The  object  of this  appearance  the  ultrastructural  experiment  o f muscle  an ocean-run  a  from  pre-rigor  muscle, several  trimmed minutes  i n a  the tissue  5  mm  i n a  blocks  chum  dehydrated  were  and  with  salmon to  allowed  at  that  1981).  (Kiernan,  phosphotungstic were  taken  acid  was  After  an ascending  The  a  then fixa-  serial d i -  i n paraffin was  dewaxed  serial  sections  Zeiss  wax. with  dilution were  haematoxylin  using  dorsal  to rest f o r  solution.  descending  sampled  the  The muscle  cut, the tissue  and water  were  from  a n d embedded  ethanol  Photographs  salmon  plate.  a  using  the physi-  parameter  excised  10% f o r m a l i n  and xylene,  sections  o f chum  was  dry petri  was  of ethanol  grades  and r e h y d r a t e d w i t h  1977).  to  microscopy  xylene,  -stained  t o observe  t h e spawning  Tissue  into  tion,  micron  contribute  and Methods  state.  f o r o n e week  Six  was  and compare t h i s  the four  fixed  lution  starvation  fish.  5.2.1. L i g h t  in  from  level  5.2. M a t e r i a l s  Muscle  and  degradation.  cal  of  maturation  MC  of  then  (Lillie, 100  light  microscope. 5.2.2. Grades a  pre-rigor  from  one  strument.  and f o u r  state.  the dorsal The  Electron  A  o f t h e chum  3 mm  muscle tissue  microscopy  diameter  of the fish was  were  core  of tissue  using  a punch  allowed  -100-  salmon  to  rest  sampled was  i n  taken  biopsy i n -  f o r  several  minutes, 7.3).  The  sodium  rinsed with  tissue  was  cacodylate  muscle M  and  was  rinsed  sodium  ethanol  fixed  buffer, in  water  and  embedded  three  Spurr's  ultramicrotome  grids.  aqueous citrate  acetate  graphed  with  5.3.  a  1981).  and  5.3.1. The stain of  the  from  the  muscle closely 5.1,  to  muscle  sections tween  used  Light  during  of  dorsal  packed,  Figure  5.2  the as  grade  cut  was  10  from  1%  were  The  OsO /0.1 4  rinsed  in in  infiltrated  and  the  20  minutes  grids  a  ultrathin  for  0.1M  dehydrated  Austria),  electron  "silver"  blocks  and  placed  minutes  with  with  with  Sato's  viewed  and  on 2%  lead photo-  microscope.  microscopy acid  haematoxylin  and 1  the  changes  clearly  spawning  fish, in  the  morphology  Micrographs  showed fish.  a  change Within  the  m y o f i b r i l s were  the  cross  longitudinal  -101-  histochemical  in  migration.  muscle  illustrated  and  hour,  1  of  stained  spawning  silver-bright  cells  for  serially  were  i n v e s t i g a t e the  the  in  tissue  in  overnight.  fixed  Ribbons  The  4°C  (pH  Discussion  phosphotungstic  was  The  for  transmission  Results  post  and  buffer  glutaraldehyde at  (Reichert,  and  cacodylate  mixture  g r i d s were  (Kiernan,  7.3,  and  Angstroms)  The  uranyl  pH  media.  an  copper  2.5%  oxide.  (500-600  U3  with  times,  sections Om  sodium  buffer  propylene  with  M  buffer  cacodylate  d i s t i l l e d  0.1  section,  section,  of bethe  quite Figure  Figure  5.3.  Figure 5.1.  Light micrograph of a s i l v e r - b r i g h t chum salmon stained with phosphotungstic acid haematoxylin. Bar represents 10 microns. (Magnification 1500). -102-  Figure 5.2.  Light micrograph of a cross section of a s i l v e r - b r i g h t chum salmon stained with phosphotungstic acid haematoxy l i n . Bar represents 20 microns. (Magnification 600). -103-  The  grade  2  and  progressively the  cells  dicate  a  section  of  tearing,  found  of  the  and  The  structural  were  were  and  the  cells  were  Thirty-six cells  observed.  the  graphs  i n two  between  grades. of  the  interesting bright  electron  micrographs  two  a  5.6  i n the  In  muscle  section  the  was  have  cells  of i n -  a  longitudinal  demonstrated  myofibrillar  light  structure.  micrographs  d e s c r i b e d  the  to  bundles.  cross  the muscle  the  spawning  and  had  also  state  had  of  salmon. lost a l l  d i s a p p e a r e d from  unstructured  i n the white  were  chinook  disorganized,  appeared  found  seen  within  Striations  Electron  Although  differences other  of  appeared  muscle,  and  swollen.  however  they  dramatic. 5.3.2.  muscle  in  extremely  show  and  who  muscle  were  spaces  the  (1926)  but  myofibril  Figure  muscle  o b s e r v e d by  cheek  changes  less  large  characteristics.  fibres,  Similar  the  5.5  in integrity  Greene  fibres  Figure  spawning  loss  myofibrillar  the  and  the  changes  myofibrils.  changes  by  Muscle  5.4  similar,  between  dramatic  i n which  loss  were  space  fish,  Figure  muscle  fish  more  spawning  cells.  3  fish  micrographic fields  grade  1  i n many the  note  exhibited  two  grade  micrographs  difficult  Figures of  and  silver-bright  i t was  muscle  to  microscopy  5.7  to  to 5.10  silver-bright  that  the  4  -104-  same  chum  there  and  different fish  were  spawning  are chum  forms  were  distinct chum,  differentiate  micrographs  many o f t h e  of  in  between  electron  micro-  salmon.  It  of  the  was  silver-  of degeneration  Figure 5.3.  Light micrograph of a longitudinal section of a s i l v e r bright chum salmon stained with phosphotungstic acid haematoxylin. Bar represents 10 microns. (Magnification 1500). -105-  Figure 5.4.  Light micrograph of a cross section of a spawning chum salmon stained with phosphotungstic acid haematoxylin. Bar represents 10 microns. (Magnification 1500). -106-  Figure 5.5.  Light micrograph of a cross section of a spawning chum salmon stained with phosphotungstic acid haematoxylin. Bar represents 20 microns. (Magnification 600). -107-  Figure 5.6.  Light micrograph of a l o n g i t u d i n a l section of a spawning chum salmon stained with phosphotungstic acid haematoxy l i n . Bar represents 10 microns. (Magnification 1500). -108-  Figure  5.7.  E l e c t r o n micrograph of the d o r s a l muscle of a s i l v e r - b r i g h t salmon. Bar on upper r i g h t corner of photograph represents 1 micron. ( M a g n i f i c a t i o n = 27,000). -109-  Figure  5.8.  E l e c t r o n micrograph of the d o r s a l muscle of a s i 1 v e r - b r i g h t salmon. Bar on upper right corner of photograph represents 1 micron. ( M a g n i f i c a t i o n = 27,000).  -110-  Figure  5.9.  E l e c t r o n micrograph of the d o r s a l muscle of a s i l v e r - b r i g h t salmon. Bar on upper r i g h t corner of photograph represents 1 micron. ( M a g n i f i c a t i o n = 27,000). - I l l -  Figure  5.10.  E l e c t r o n micrograph of the d o r s a l muscle of a s i 1 v e r - b r i g h t salmon. Bar on upper right corner of photograph represents 1 micron. ( M a g n i f i c a t i o n = 27,000). -112-  as  the  As  a  spawning  result,  a  by  of  For  assigning a  each  teria  the  photograph  was  were  the  partial  and  myosin  score  of  or  tears  tears  Inter-myofibril space  normally  T-tubules spaces was  were  between  assigned  even  were  using  unusual.  Thinning,  the  of the  score judges  of  the  to  and  1  Figure  distin5.12, If  a  any  assigned. in  the  Swollen 1.  a  If  score  the of  swollen.  5.14,  the  actin  Figure  of  determine  in  of  be  was  regular,  within  5.11,  increase  score  cri-  crite-  f i b r i l s .  Figure  myosin  4  bundles.  an  score  The  could  appeared  myofilaments  actin  a  and  tubules  considered  between  Five  the  was  for  small  4.  pattern  of  muscle  was  density  a  i f  were  score  5.13,  sufficient  final  Splitting,  myofibril a  between  spaces  assigned  observed,  bundles  myosin  scored  of, t h e  to  distinct  and  the  found  photograph  spaces  within  sum  0  was  1  The  the  the  f i s h  were  Smearing,  assigned.  Figure  not  as  or  .  between  a  actin  spaces,  entire  creased  If  of  between  criteria  observed),  follows.  loss  0 was  were  four  ranged  frequently.  spawning  photograph)  as  complete  filaments.  a  or  scores  less  distinguish  calculated  described  or  breaks  breaks  the  (none  much  and  micrograph,  on  used  were  each  somewhere  ria  to  s i l v e r - b r i g h t  value  thus  present  criteria  0  and  guished,  of  were  of  (observed for  but  list  micrographs developed.  fish,  0  The  whether  the  was  de-  the  fibril.  Small  myofibrils  were  1.  were  asked  s c o r i n g system,  and  to  categorize  their  -113-  results  the are  micrographs given  in  Figure  5.11.  E l e c t r o n m i c r o g r a p h of the d o r s a l muscle of s p a w n i n g chum s a l m o n . "Smearing" i s present on t h e u p p e r p o r t i o n o f t h e p h o t o g r a p h . Bar on upper r i g h t c o r n e r o f p h o t o g r a p h represents 1 micron. ( M a g n i f i c a t i o n = 26,000). -114-  Figure  5.12.  E l e c t r o n micrograph of the d o r s a l muscle of s p a w n i n g chum s a l m o n . " S p l i t t i n g " i s present to the r i g h t of the photograph. B a r on u p p e r right corner of photograph represents l micron. ( M a g n i f i c a t i o n = 26,000). -115-  Figure  5.13.  E l e c t r o n micrograph of the d o r s a l muscle of s p a w n i n g chum s a l m o n . Notice areas with "inter-myofibril spaces" B a r on u p p e r r i g h t corner of photograph represents 1 micron. ( M a g n i f i c a t i o n = 26,000). -116-  Figure  5.14.  E l e c t r o n micrograph of the d o r s a l muscle of s p a w n i n g chum s a l m o n . "Thinning" within the m y o f i b r i l bundles i s apparent. B a r on u p p e r right corner of photograph represents l micron. ( M a g n i f i c a t i o n = 26,000).  -117  Table  5.1.  signing zero,  The  the  or  ever,  one.  I t was  as  reflection  of  areas  bright that  of  number  of of  system  could  mature  fish.  the  between  the of  chum  the  order It  to  system  integrity  not  have  to  correctly  assess  successfully  distinguish  -118-  not  a  between  in  any  the  silver  to  assume  areas  consider  fish,  a  which  a  degree  this mature  of  number  therefore, that  same  be  degraded, of  would  the  How-  areas  severely  necessary  and  i t was  some  correct  from  to  should  not  micrographs  possible  degradation  Although  i s expected  of  Instead,  muscle  and  as-  score  field.  i t was  fish  I t was  fish.  microscopic  appeared  structure  not  in  silver-bright  method. of  rate  cumulative i t was  grading the  success  the  single  Conversely,  in  a  maturation.  silver-bright  micrographs  87%  that  nature  spawning  degradation.  maturity a  sexual  to  samples  a  of  focal  retained  sample.  the  muscle  of  on  failure  the  during  muscle  other  based  a  72  concluded  inability  considered  the  a  distinguish  samples  this  occured  had  silver-bright  consistently spawning  judges  of  given  scoring and  im-  Table  5.1.  Percentage of silver-bright and spawning fish a s s i g n e d t o each s c o r e used t o judge micrographs o f chum d o r s a l muscle.  Grade Judge  0  1  Score 2  Total 3  4  1  1  Silver Spawn  28 1  13 3  6 13  3 19  0 12  87%  2  Silver Spawn  20 6  20 9  3 10  9 11  0 11  77%  3  Silver Spawn  10 0  19 3  19 13  7 19  0 8  72%  4  Silver Spawn  22 5  22 10  12 10  0 17  0 2  73%  5  Silver Spawn  7 3  44 17  5 12  0 12  0 0  75%  Total correct classification. Silver-bright fish w e r e c l a s s i f i e d a s h a v i n g s c o r e s l e s s t h a n 2.  -119-  6 . CONCLUSIONS  The keta) of  spawning  adversely  the  canned  i n the  product,  order the  affected  product.  silver-brights final  to  are  of  the  chum  ocean  would  stocks  harvested  as  improve  prevent of  they  fish,  (Oncorhynchus  and  eating  harvesting  problems  weaker  salmon  appearance  Although  other  protect  chum  migration  the  quality  chum  only  as  the  q u a l i t y of  the  this  practice.  In  a  approach  large the  portion  mouth  of  of the  river. The chemical  object and  throughout using  of  this  textural  their  methods  such  as  assays,  copy.  addition,  method  were  The  used  fish  their  external  slime  layer  tion  of  Hunter tion based  Lab  of on The  to  a  light  and  graded  into  i t s absorbance visual  white-grey  and  i n the  change  at in  colour  This  physical,  in  was  the  analysis,  electron an  chum  accomplished  colour  and  micros-  instrumental  skin  categories colour,  considerations the  colourimeter,  four  The  important of  occurred  analysis,  panels  the  texture.  appearance.  Colour  assess  transmission  sensory  were  astaxanthin  which  proximate  measure  grade.  to  migration.  to  were  was  changes  spawning  proteinase In  work  raw by  flesh  thickness  in  was  calculating  the  the  and  with  a  concentrathe  muscle  red-pink  colour  nm.  the  was  of  on  determina-  measured  fat-soluble portion 475  based  muscle a  -120-  from  reflection  a  of  the  loss  of  astaxanthin values. with  in  The  the  muscle  content  of  tion  a n a l y s i s between  that  a/L,  tively was  ratio  there  were  15.8%  and 74%  strated muscle  as  from  79%  as  the  grade  =  (a  during  second f i r s t with  bite  fish  The ability the  the  and  the  two  colour  The  best  Lab  significantly  values  also  de-  Correla-  values  indicated  significantly indicator  discrimination  and  nega-  of  grade  of  the  in  1) 2,  2 decreased  the  decreased between  were  to  moisture demon-  energy  sources  in  the  not  ef-  the and  grade  (TPA)  was  a  maximum  salmon  the  in  work  done  on  the  slope  of  the  significantly  grade.  more  successful  grades  was  of  work  increased  the  difference  ratio  the  the  significantly  -121-  %  results  detect  with  four  each  in  softening of  to  Cohesiveness  panelists  17.9  These  analysis  (Area slope  from  that  i t s migration.  progressive  and  findings  increases  matured.  profile  bite  distinguish  attributes  in protein,  cohesiveness,  2),  slope  literature  parameters  first  (Area  fish  texture  were  sensory to  b  Hunter  increased.  were  1.0%  progresses  only  0.05)  to  depletion  compression. grade  and  a/b  confirmed  4.9%  i n measuring The  a  the  decreases  the  severe  muscle.  and  the  decreased  value  and  grade.  analysis  Instrumental fective  L  in  a/L.  lipid  the  the  'a',  significant  to  changes  Hunter  grade  astaxanthin,  Proximate  done  but  c o r r e l a t e d with  the  from  The  significantly,  the  astaxanthin  i n c r e a s i n g grade.  creased  and  of  salmon.  with most  grade,  in  their A l l  of  however  successful  with  the  s e n s o r y  a t t r i b u t e  Z-transformations ble  on t h e data  thus  the analysis  distinguishing relation  texture  that  between  analysis  negatively  sures  t h e grades  indicated correlated  with  best  correlation  indicated  that  correlated  t h e t e x t u r e o f t h e canned  of  myofibrillar  change.  Because  particular was  proteins,  strength. suring  the force  across  each  result  i n a  loss  the moisture  more  fibres  attribute.  was  was a d e f i n i t e  however  presented the loss that  of  would  also  -122-  slide  salmon  would  amount  increase  of  connec-  collagen,  mois-  increasing  to slide  increase.  mea-  to  and an  of gelatinized  necessary  i n any  myofibrillar  fibres  proteins  and t h e r e l a t i v e  the force  this  t h e I n s t r o n was  and s a r c o p l a s m i c p r o t e i n s i n c r e a s e d w i t h  other  mea-  to the Instron,  ture  across  The  softening  not oriented  f o r t h e muscle  As t h e c o n t e n t  fibres  and  by t h e breakdown  tissue.  each  significantly  attributes,  tive  of the fish,  measures  cohesiveness.  caused  i n Cor-  and instrumental  were  the myofibrillar  turity  between  effective  Starvation i n the migrating  content  possi-  t h e I n s t r o n was n o t m e a s u r i n g  necessary  other.  there  t o measure  I t i s possible  was  sensory  sensory  i n t h e sample  not possible  of variances  with  product  t h e muscle  direction  i t was  and instrumental  the four  although  p e r f o r m i n g  judge,  cohesiveness  between  that  By  f o r each  sensory  of  in  each  of variance  between  fibrousness  negative  it  from  t o e l i m i n a t e t h e non-homogeneity  judges  of  d r y n e s s .  ma-  t h e muscle  The pH  proteinase activity  3.5,  6.2  a c t i v i t y  and  with  phosphatase, cantly  visible  crographs number tions ing,  graphs This  loss  of  the  smearing,  a  myofibrils,  and  mine  a  the  maturity  the  focal  graphs  of  apparent  loss  muscle  the fish.  of  nature  a  of  excess  of  of to  micrograph  was  i t was on  one  also  -123-  Light  myofibrillar  focal  including  bundles,  helped  cell,  deter-  spawning  photograph  to  some  or  predict  because  addition,  demonstrated  the  myofilaments  possible  In  of  i n the  the  a  sec-  micro-  organization  of  or  degeneration.  criteria  from not  mi-  tear-  Electron  criteria  degeneration. fish  observe  in myofibril  density four  based  the  to  Longitudinal  of  four  these  silver-bright  useful  diameter,  cytoplasmic f l u i d  i n the  the  be  used  the  muscle.  areas  or  However,  of  the  breaks  fish  in  occurrence  tears  of  possible  degradation  muscle.  cells.  loss  Use  acid  s i g n i f i -  the  would  the  complete  decease  fibril.  silver-bright  of  by  or  the  were  described  spaces,  whether  increased  work  micrographs  revealed  was  of  enzymes.  i n the  splitting,  a  activity  also  structure  an  partial  thinning,  within  these  muscle  intermyofibril  The  Further  integrity  degeneration  in  to  increased  of  increase  enzymes  of myofibrils  and  significant  lysosomal  i n the  an  at  indicated  revealed  showed  measured  results  electron  changes  was  3.5.  marker,  migration.  and  a  muscle  These  identify  Light  the  was  pH  lysosomal  of  during and  at  grade.  contribution  isolate  There  grade  a  with  muscle  7.0.  of  of  microof  the  same c h a n g e s , to the  look  but to a lesser  a t a number  degree.  o f photographs  fish.  -124-  I t would  to accurately  be  necessary  distinguish  7. 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APPENDIXES  Absorbance a t U.V. 280 o f g r a d e as a m e a s u r e o f a c t i v i t y o f chum d o r s a l m u s c l e  s o l u b l e N versus t h e p r o t e i n a s e a t pH 3 . 5 .  0.30  0.20 O oo CM  > ZD  <D O c  0.10  CO _Q i_  O  U) j Q  < 0.00  -0.10 ' — ; 1  1  1  2  3  3  - Index of Maturity  -134-  4  4  4  Table  8.2.  Absorbance a t U.V. 280 of s o l u b l e N versus grade as a measure o f t h e p r o t e i n a s e a c t i v i t y o f c h u m d o r s a l m u s c l e a t pH 6.2.  0.30  0.22 O 03 OJ  > D  0.14  0) u  c  (rj  -Q i_ O  0.06  CO  n <  -0.02  -0.10 ' 1  1  ' 1 2 3 3 4 Index of Maturity  -135-  4  4  Table  8.3.  Absorbance a t U.V. 280 of s o l u b l e N versus grade as a measure o f the p r o t e i n a s e a c t i v i t y o f c h u m d o r s a l m u s c l e a t pH 7.0.  0.30  0.22  h  O 00  CM  >  0.14  0)  o c  iTj  n o (f)  0.06  <  -0.02  h  -0.10 1  1  1  2  3  3  4  4  Index of M a t u r i t y .  -136-  4  T a b l e 8.4.  C o n c e n t r a t i o n o f p h o s p h a t e (mM) v e r s u s g r a d e as a measure o f t h e a c i d p h o s p h a t a s e a c t i v i t y of chum muscle d u r i n g spawning m i g r a t i o n .  0.46  OJ  1  1  1  2 Index  -137-  2  3 of  3  Maturity  4  4  4  


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