Pigment Reduction in Corn Gluten Meal and Its Effects on Muscle ...

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this trial was to evaluate the effect of <strong>in</strong>creas<strong>in</strong>g levels of CGM (up to 18%) on growth <strong>and</strong>pigment deposition <strong>in</strong> fillets from ra<strong>in</strong>bow trout.4.2 Materials <strong>and</strong> methods4.2.1 Fish husb<strong>and</strong>ry <strong>and</strong> experimental conditionsThis study was conducted at the Alma Aquaculture Research Station (AARS, Alma, ON,Canada). Domestic stra<strong>in</strong> of ra<strong>in</strong>bow trout, (O. mykiss) from the AARS’s stock population wereadapted to experimental conditions for two weeks prior to start the experiment. Dur<strong>in</strong>g thatperiod, fish were fed a commercial (with no pigment supplementation) trout diet (Mart<strong>in</strong> MillsInc, Elora, ON, Canada) once a day.The 7 experimental diets were r<strong>and</strong>omly assigned to each of 21 (1,500 L) fibreglass tanks(n=3). Seventy five fish (549 g fish -1 , <strong>in</strong>itial body weight) were r<strong>and</strong>omly allocated <strong>in</strong>to eachtank <strong>and</strong> kept under a 12 h light: 12 h dark photoperiod regime. Filtered well water was suppliedapproximately at a 30.5 L m<strong>in</strong> -1 rate. Water temperature was kept at 8.5 °C. This experiment wasconducted accord<strong>in</strong>g with the guidel<strong>in</strong>es stated by the Canadian Council on Animal Care(CCAC, 1984) <strong>and</strong> the University of Guelph Animal Care Committee.Fish were h<strong>and</strong>-fed twice a week to near satiation dur<strong>in</strong>g 24 weeks. Dur<strong>in</strong>g the rema<strong>in</strong><strong>in</strong>gfive days, fish were fed us<strong>in</strong>g belt feeders. Daily ration to be dispensed by belt feeders wasdeterm<strong>in</strong>ed by averag<strong>in</strong>g the amount of feed given to that tank dur<strong>in</strong>g the previous two dailyh<strong>and</strong> feed<strong>in</strong>gs.Five fish were sampled at the beg<strong>in</strong>n<strong>in</strong>g of the experiment for the determ<strong>in</strong>ation of <strong>in</strong>itialcarcass composition <strong>and</strong> another sample of five fish was used for <strong>in</strong>itial muscle pigment content<strong>and</strong> colour determ<strong>in</strong>ation. At the end of the experimental period, four <strong>and</strong> ten fish from each tank77
were r<strong>and</strong>omly sampled for f<strong>in</strong>al carcass composition <strong>and</strong> f<strong>in</strong>al muscle pigment content <strong>and</strong>colour determ<strong>in</strong>ation, respectively. Fish were sacrificed by an overdose of MS-222 (300 ppm L -1 ). Fish for carcass composition determ<strong>in</strong>ation were autoclaved, thoroughly ground <strong>in</strong>to a slurry,freeze-dried, ground <strong>in</strong>to a f<strong>in</strong>e powder <strong>and</strong> kept at -20 °C until analysis. Fish to be analysed formuscle pigment content <strong>and</strong> colour determ<strong>in</strong>ation were manually sk<strong>in</strong>ned <strong>and</strong> right h<strong>and</strong> sidefilleted right after slaughter; after muscle colour determ<strong>in</strong>ation muscle samples were stored at -20 °C until pigments determ<strong>in</strong>ation analysis.4.2.2 Experimental dietsSix (6) extruded experimental diets were formulated to be isonitrogenous (48 % crudeprote<strong>in</strong>) <strong>and</strong> isoenergetic (20 MJ DE/kg) <strong>and</strong> to conta<strong>in</strong> different <strong>in</strong>gredient comb<strong>in</strong>ations (i.e.fish meal, soy prote<strong>in</strong> concentrate, feather meal, blood meal, wheat meal, soybean meal <strong>and</strong> fishoil) <strong>and</strong> either no CGM (Diet 1), 4% CGM (Diet 2), 9% CGM (Diet 3), 12% CGM (Diet 4), 14%CGM (Diet 5) <strong>and</strong> 18% CGM (Diet 6), <strong>and</strong> meet all the nutritional requirements for ra<strong>in</strong>bowtrout based on NRC (2011) (Table 4.1). A commercial feed conta<strong>in</strong><strong>in</strong>g 10% CGM (Diet 7) wasused. All diets were supplemented with 50 mg kg -1 astaxanth<strong>in</strong> (Carophyll® p<strong>in</strong>k, 8%astaxanth<strong>in</strong>, DSM Nutritional Products, ON, Canada).Noteworthy is that <strong>in</strong>clusion of graded levels of CGM <strong>in</strong> Diet 1, 3 <strong>and</strong> 6 (0, 9 <strong>and</strong> 18%CGM, respectively) was achieved at expense of soy prote<strong>in</strong> concentrate only (Table 4.1).Conversely, <strong>in</strong> the formulation for Diets 2, 4 <strong>and</strong> 5 (4, 12 <strong>and</strong> 14% CGM, respectively), <strong>in</strong>clusionof several <strong>in</strong>gredients (i.e. fish meal, feather meal, blood meal, wheat meal <strong>and</strong> fish oil) wasmodified <strong>in</strong> order to accomplish desired CGM <strong>in</strong>clusion levels.78
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Pigment Re
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ingredients. Furth
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Thanks to all my colleagues, friend
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2.10.1 Enzymatic treatments .......
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LIST OF TABLESTable 3. 1 - Factors
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LIST OF FIGURESFigure 2. 1 - Molecu
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Ingredients commonly in</st
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CHAPTER - 2 LITERATURE REVIEW2.1 In
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leached when 5% of soy flour as LOX
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deep and fast glyc
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observed in ra<str
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Chylomicron constituents are remove
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Muscle growth in r
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2.7.2 Corn gluten
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2010). However, due to its imbalanc
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No differences in
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supplemented with 42% of a vegetabl
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Soybean seeds are the richest known
Page 37 and 38: 2.10.1.4 Bleaching
Page 39 and 40: 2.11 ConclusionInclusion of <strong
Page 41 and 42: 4.0%*68%* 5.6%* 12%* 3.5%* 6.5%*Fig
Page 43 and 44: Figure 2. 4 - Schematic representat
Page 45 and 46: 3.1 IntroductionThe characteristic
Page 47 and 48: many factors such as LOX source lev
Page 49 and 50: During the 12-week
Page 51 and 52: 3.2.3 Analytical methodsDry matter
Page 53 and 54: solution was generated by mix<stron
Page 55 and 56: 3.2.4 Statistical analysisData from
Page 57 and 58: Factors defined as
Page 59 and 60: chromatography). Relative amounts o
Page 61 and 62: After carotenoids are absorbed thro
Page 63 and 64: and ended up <stro
Page 65 and 66: Table 3. 2 - Experimental design us
Page 67 and 68: Table 3. 4 - Analysed proximate com
Page 69 and 70: Proximate composition (Analysed; dr
Page 71 and 72: Table 3. 7 - Lipoxygenase activity
Page 73 and 74: 18 1 0 -1 0 120 15 0 10 2519 -1 0 1
Page 75 and 76: Table 3. 10 - HPLC carotenoid profi
Page 77 and 78: Table 3. 12 - Retain</stron
Page 79 and 80: Table 3. 14 - Fillet carotenoid con
Page 81 and 82: Total yellow pigment(mg•kg -1 )25
Page 83 and 84: Figure 3. 4 - Growth curve of ra<st
Page 85 and 86: effects of dietary CGM and<
Page 87: soybean meal in a
Page 91 and 92: An aliquot (1 ml) of chloroform low
Page 93 and 94: Retained nitrogen
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Page 97 and 98: Values for all-trans astaxanth<stro
Page 99 and 100: previous reports where colour attri
Page 101 and 102: Proximate composition (analysed), d
Page 103 and 104: Table 4. 2 - Growth performance of
Page 105 and 106: Table 4. 3 - Carcass chemical proxi
Page 107 and 108: Table 4. 4 - Retain</strong
Page 109 and 110: Table 4. 5 - Pigment</stron
Page 111 and 112: Table 4. 6 - Colour attributes <str
Page 113 and 114: FiguresFigure 4. 1 - Growth curves
Page 115 and 116: Figure 4. 3 - Muscle colour attribu
Page 117 and 118: the effects of dietary reduced-caro
Page 119 and 120: astaxanthin deposi
Page 121 and 122: After steeping, ke
Page 123 and 124: 0.9994 and 0.9999,
Page 125 and 126: under acidic and a
Page 127 and 128: oxidation within t
Page 129 and 130: colour difference usin</str
Page 131 and 132: TablesTable 5. 1 - Analyzed proxima
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Effects of
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Data are mean (n=2).a Steepwater pH
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CHAPTER - 6 GENERAL DISCUSSIONCurre
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significant reduction in</s
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Reduction of pigme
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though wheat gluten meal conta<stro
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BibliographyAas, G.H., Storebakken,
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Bjerkeng, B., Hatlen, B., Wathne, E
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Choubert, G., Cravedi, J., Laurenti
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Fauconneau, B., Andre, S., Chmaitil
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Johnston, I., Li, X., Vieira, V., N
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Li, M.H., Oberle, D.F., Lucas, P.M.
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Peter, C.M., Han, Y., Bolin
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Skonberg, D.I., Hardy, R.W., Barrow
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Ytrestoyl, T., Struksnæs, G., Kopp
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