inclusion of camelina meal as a protein source in diets ...€¦ · rainbow trout (oncorhynchus...

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1 Department of Ocean Sciences, Memorial University of Newfoundland, St. John’s, Newfoundland and Labrador, Canada;2 Faculty of Agriculture, Dalhousie University, Truro, Nova Scotia, Canada

Camelina meal (Camelina sativa) (CM) is a potential pro-

tein source for aquaculture feeds, on account of its crude

protein level (380 g kg�1) and inclusion of most indispens-

able amino acids. Two experiments were conducted with

rainbow trout (Oncorhynchus mykiss) and Atlantic salmon

(Salmo salar). Rainbow trout (44.9 g fish�1) were fed diets

with CM at 0 g kg�1 (0% CM), 70 g kg�1 (7% CM),

140 g kg�1 (14% CM) or 210 g kg�1 (21% CM) for

12 weeks at 14 °C in freshwater, and salmon

(241.8 g fish�1) were fed diets with CM at 0 g kg�1 (0%

CM), 80 g kg�1 (8% CM), 160 g kg�1 (16% CM) or

240 g kg�1 (24% CM) for 16 weeks at 14 °C in sea water.

Growth, lipid and amino acid tissue compositions were

compared between species. Trout could tolerate up to 14%

CM diets without affecting the growth compared to the

control, while salmon fed ≥8% CM gained less weight than

the control (P = 0.008). The feed conversion ratio in trout

fed 21% CM was higher than the control (P = 0.002), and

feed intake in salmon fed ≥8% CM was lower than the

control (P = 0.006). Trout fatty acid and amino acid

composition showed minimal differences between CM-fed

and control-fed fish, while salmon showed significant alter-

ations after feeding CM diets. Multivariate analyses

emphasized differences in tissue composition between spe-

cies fed CM diets.

KEY WORDS: amino acids, Atlantic salmon, camelina, fatty

acids, fish meal, rainbow trout

Received 26 June 2014; accepted 30 November 2014

Correspondence: S.M. Hixson, Department of Ocean Sciences, Memorial

University of Newfoundland, 1 Marine Lab Road, St. John’s, Newfound-

land, Canada A1C 5S7. E-mail: [email protected]

Fish meal (FM) is a finite resource, and predictions estimate

that the availability and cost of FM will pose serious pres-

sure on the aquaculture industry in the next 5 years. The

focus of nutritional research in aquaculture in the past

15 years has been on finding sustainable and economical FM

alternatives and testing their efficacy towards the production

of various farmed fish species. In recent years, the commer-

cial feed industry has reduced their dependence on marine

ingredients in an effort to achieve sustainable growth by

including a number of different plant and animal protein

sources (Crampton & Carr 2012). Plant meals are mainly

supplied in the diet as a protein source, but also have neces-

sary functional properties for proper pellet quality. There-

fore, plant ingredients are chosen based on protein quality,

functionality, availability and cost, and these raw materials

can act as substitutes for one another (Crampton & Carr

2012). A single ingredient cannot totally replace FM, but a

mixture of ingredients can mimic the amino acid profile of

FM (Li et al. 2009). New alternative proteins must be tested

to decrease the pressure on FM and provide alternative

options for combinations of protein sources in fish feeds.

The oilseed camelina (Camelina sativa) is a member of

the Cruciferae (Brassicaceae) family, which includes mus-

tards, rapes, broccoli, cabbage, collards, cauliflower and

many weeds. This robust plant requires minimal input for

growth, grows well in low-fertility and saline soils, is toler-

ant of insects and can survive frost and freeze–thaw cycles

after emergence during late winter and spring (Putnam

et al. 1993). Camelina oil is considered a potential source

for biofuels, particularly jet fuel (Mudalkar et al. 2014), as

well as a lipid source in aquaculture feeds for rainbow

trout (Oncorhynchus mykiss) (Hixson et al. 2014a), Atlantic

salmon (Salmo salar) (Hixson et al. 2014b) and Atlantic

cod (Gadus morhua) (Morais et al. 2012). Therefore, there

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ª 2015 John Wiley & Sons Ltd

2015 doi: 10.1111/anu.12276. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Aquaculture Nutrition

was considerable interest in the remaining meal by-product

available after oil extraction. Camelina meal (CM) is con-

sidered a potential protein source in aquaculture feeds, on

account of its crude protein level (380 g kg�1) and inclu-

sion of some indispensable amino acids such as methionine,

lysine, phenylalanine, threonine, leucine, isoleucine and

valine. The lipid fraction remaining in the meal (50 g kg�1)

is high in n-3 polyunsaturated fatty acids (18:3n-3), n-6

fatty acids (18:2n-6) and monounsaturated fatty acids. The

CM used in this study was solvent-extracted to resemble

the oilseed by-product obtained if most of the oil was

removed for other purposes such as biofuel.

Generally, alternative plant protein sources can be

included in diets at relatively low levels without compro-

mising growth. However, including higher levels of these

protein sources in diets for different fish species, particu-

larly carnivorous fish, has reduced feed intake, prevented

digestion and nutrient absorption, induced gut inflamma-

tion, decreased intestinal mucosal length and slowed

growth (Krogdahl et al. 2010). CM contains secondary

plant metabolites such as glucosinolates, sinapines, tannins

and phytate, which are antinutritional factors and may

induce physiological effects if present in significant levels

(Matthaus 1997). Therefore, diets must be carefully formu-

lated to balance protein and amino acids from plant pro-

teins and FM so that fish health, growth, environmental

sustainability and cost efficiency can be achieved.

The objective of this study was to determine the level of

CM, which can be included in the diet without affecting

the growth in rainbow trout and Atlantic salmon. Graded

levels of CM were included up to 210 g kg�1 (rainbow

trout) and 240 g kg�1 (salmon) in the diet to determine the

level at which growth, tissue lipid and fatty acid composi-

tion, and tissue amino acid composition are affected.

Another purpose of the study was to determine whether

amino acids and fatty acids in CM were correlated with

changes in growth using multivariate statistics and also

whether dietary CM would affect the sensory quality of the

final product. This is the first study to use graded levels of

CM to determine an optimal level in diets for salmonids.

Camelina (Calena cultivar) was grown and harvested at

Dalhousie University’s off-campus location (Canning, NS,

Canada). Seeds were single-pressed using a KEK 0500

press at Atlantic Oilseed Processing, Ltd. (Summerside, PE,

Canada) to first extract the oil. The remaining meal was

ground with a hammer mill (screen size 8 mm) into a pre-

pressed meal cake at Atlantic Oilseed Processing, then sol-

vent-extracted with petroleum ether at a concentration of

3 mL g�1 at the Faculty of Agriculture Campus, Dalhousie

University (Truro, NS, Canada). All diets were formulated

as isonitrogenous and isocaloric practical diets and were

produced at the Faculty of Agriculture Campus, Dalhousie

University. In the first experiment with rainbow trout, the

dietary treatments were the following: a control diet with

0 g kg�1 CM (0%), and experimental diets containing 70 g

kg�1 CM (7%), 140 g kg�1 CM (14%) and 210 g kg�1

CM (21%). In the following experiment for Atlantic sal-

mon, dietary treatments included slightly higher increments

of CM: a control diet with 0 g kg�1 CM (0%), and experi-

mental diets containing 80 g kg�1 (8%), 160 g kg�1 (16%)

and 240 g kg�1 (24%). Diets were formulated to meet

nutritional requirements for salmonids (NRC 2011). The

diet formulations were based on the published values for

digestible protein and energy of ingredients (NRC 2011)

and the proximate composition and nutrient digestibility

values of solvent-extracted CM used in the present study

(Fraser & Anderson 2012; Ye 2014). Digestibility experi-

ments with rainbow trout and Atlantic salmon (Fraser &

Anderson 2012) were performed before growth experi-

ments; therefore, ‘digestible nutrients’ values for the CM

used in this study were available for the formulations in

this study. All diets were steam-pelleted using a laboratory

pelleting mill (California Pellet Mill, San Francisco, CA,

USA). Diets were stored at �20 °C until needed.

An experiment was conducted with juvenile rainbow trout

(44.9 � 10 g fish�1 mean initial weight � SD; 15.7 �1.2 cm mean initial length) at the Faculty of Agriculture

Campus, Dalhousie University. Fish were received from Fra-

ser Mill’s hatchery (Antigonish, NS, Canada). Ethical treat-

ment of fish in this experiment followed guidelines of the

Canadian Council of Animal Care (Dalhousie University

Animal Care approved, 2011-016). Fish were randomly dis-

tributed (1116 total) into 12 experimental tanks (200 L),

each tank with 93 fish. Dietary treatments were administered

to triplicate tanks for the 12-week trial. The fish were accli-

mated on the control diet for 1 week prior to initial sam-

pling. A flow through system of freshwater was supplied to

each tank at a rate of 10 L min�1 and a photoperiod of

12 h. The dissolved oxygen (10 mg L�1) and water temper-

ature (14 °C) were monitored daily. Fish were fed to

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Aquaculture Nutrition ª 2015 John Wiley & Sons Ltd

apparent satiation twice daily (0900 and 1600), and feed

consumption was recorded weekly. The initial size of the

pellet was 1.5 mm and increased to 2.5 mm as the fish grew

larger throughout each trial. Mortalities were weighed and

recorded.

A second experiment was conducted with salmon smolts in

sea water (242 � 46 g fish�1 mean initial weight � SD;

27 � 1.8 cm mean initial length) at the Ocean Sciences

Centre (Memorial University of Newfoundland, St. John’s,

Newfoundland and Labrador, Canada). Fish were received

from Cooke Aquaculture (St. Alban’s, Newfoundland and

Labrador, Canada). The salmon (Saint John River stock)

were transferred from the freshwater hatchery to the Ocean

Sciences Centre’s Joe Brown Aquatic Research Building

(JBARB) to undergo smoltification in sea water. Ethical

treatment of fish in this experiment followed guidelines of

the Canadian Council of Animal Care (Memorial University

Institutional Animal Care Protocol approved, 12-50-MR).

The smolts were randomly distributed (600 total) into 15

experimental tanks (500 L), each tank with 50 fish. Dietary

treatments were administered to triplicate tanks for the

16-week trial. Salmon were fed an additional 4 weeks than

rainbow trout to further test the results obtained from the

previous experiment. The fish were acclimated on the control

diet for 1 week prior to initial sampling. Throughout the

duration of the trial, a flow through system of filtered sea

water was supplied to each tank at a rate of 12 L min�1 and

a photoperiod of 12 h. The dissolved oxygen (10 mg L�1)

and water temperature (14 °C) were monitored daily. Fish

were fed to apparent satiation twice daily (0900 and 1600).

The initial size of the pellet was 4.0 mm; it increased to

6.0 mm as the fish grew larger. Mortalities were weighed and

recorded.

Rainbow trout sampling occurred at the Faculty of Agri-

culture Campus, Dalhousie University at week 0 (the day

before experimental diets were fed) and week 12. Atlantic

salmon sampling occurred at the Ocean Sciences Centre at

week 0 (the day before experimental diets were fed) and

week 16. Individual fish were rapidly netted and eutha-

nized by an overdose of anaesthetic (buffered tricaine

methane sulphonate, TMS), and clinical signs of death

were ensured prior to sampling. Three fish per tank were

randomly sampled on each sampling date and measured

for length and weight. The skin was removed on the left

side, and the white muscle tissue was subsampled for

analyses.

Lipid samples were stored on ice during sampling of

each tank and were processed within an hour. Samples

were collected in 50-mL test tubes that had been rinsed

three times with methanol followed by three rinses with

chloroform. The tubes were allowed to dry completely

before they were weighed. The tubes were weighed again

following the addition of the sample. After wet weights of

samples were recorded, samples were covered with 8 mL

of chloroform, the headspace in the tube was filled with

nitrogen, and the Teflon-lined caps were sealed with

Teflon tape and stored at �20 °C. The rainbow trout

samples were stored at �20 °C until shipment to the

Ocean Sciences Centre for analysis (CCAC 12-09-MR,

approved protocol for use of fish tissues from Dalhousie

University). Amino acid samples and carbon, hydrogen

and nitrogen samples were also stored on ice during sam-

pling of each tank and were processed within an hour.

Samples were collected in 20-mL scintillation vials that

had been heated in a furnace for 24 h at 450 °C to burn

the organic material. The vials were weighed before and

after the samples were added. The samples were stored at

�20 °C until analysis.

Lipid samples were extracted according to Parrish (1999).

Samples were homogenized with a Polytron PCU-2-110

homogenizer (Brinkmann Instruments, Rexdale, ON, Can-

ada) in a 2 : 1 mixture of ice-cold chloroform: methanol.

Chloroform-extracted water was added to bring the ratio

of chloroform : methanol : water to 8 : 4 : 3. The sample

was sonicated for 4 min in an ice bath and centrifuged at

2688 RCF (Relative Centrifugal Force) for 2 min at room

temperature. The bottom, organic layer was removed

using a double pipetting technique. A pipette (1 mL) was

placed inside the test tube with negative air pressure to

pass through the top aqueous phase. The tip of the 2-mL

pipette was then placed inside the 1-mL pipette to draw

the bottom organic layer up through the top layer with-

out disturbing the interphase. The organic layer was col-

lected and pooled in a separate test tube. Chloroform

was then added back to the extraction test tube, and the

entire procedure was repeated three times for muscle sam-

ples and five times for liver samples. The samples were

concentrated using a flash-evaporator (Buchler Instru-

ments, Fort Lee, NJ, USA).

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Lipid class composition was determined using an Iatroscan

Mark VI TLC-FID (Mitsubishi Kagaku Iatron, Inc., Tokyo,

Japan), silica-coated Chromarods and a three-step develop-

ment method (Parrish 1987). The lipid extracts were applied

to the Chromarods and focused to a narrow band using ace-

tone. The first development system was hexane : diethyl

ether : formic acid (98.95 : 1.0 : 0.05). The rods were devel-

oped for 25 min, removed from the system for 5 min and

replaced for 20 min. Each rod was then scanned to the low-

est point behind the ketone peak. The second development

was for 40 min in hexane : diethyl ether : formic acid

(79 : 20 : 1), and then, rods were scanned to the lowest point

behind the diacylglycerol peak. The final development sys-

tem had two steps: the first was 100% acetone for two 15-

min time periods, followed by two 10-min periods in chloro-

form : methanol : chloroform-extracted water (5 : 4 : 1);

then, the rods were completely scanned. Before using each

solvent system, the rods were dried in a constant humidity

chamber. The data were collected using Peak Simple soft-

ware (version 3.67; SRI Inc., Menlo Park, CA, USA). The

Chromarods were calibrated using standards from Sigma

Chemicals (St. Louis, MO, USA).

Lipid extracts of tissues were transesterified using the Hil-

ditch reagent (1.5 H2SO4 : 98.5 anhydrous methanol) for

1 h at 100 °C. Reagents were added in the proportion of

1.5 mL reagent per 4–16 mg of lipid (Morrison & Smith

1964). To check the derivatization efficiency, samples were

transesterified and then the lipid class composition of the

methyl ester solution was determined by thin layer chroma-

tography with a flame ionization detector.

All FAMEs were analysed on a HP 6890 gas chromato-

graphy with a flame ionization detector (GC-FID), equipped

with a 7683 autosampler. The column was a ZB wax+ (Phe-

nomenex, Torrance, CA, USA). The column length was

30 m with an internal diameter of 0.32 mm. The column

temperature began at 65 °C where it was held for 0.5 min.

The temperature ramped to 195 °C at a rate of

40 °C min�1, held for 15 min and then ramped to a final

temperature of 220 °C at a rate of 2 °C min�1. This final

temperature was held for 45 s. The carrier gas was hydrogen

flowing at 2 mL min�1. The injector temperature started at

150 °C and ramped to a final temperature of 250 °C at

120 °C min�1. The detector temperature stayed at 260 °C.

Peaks were identified using retention times from standards

purchased from Supelco (Bellefonte, PA, USA): 37 compo-

nent FAME mix (Product number 47885-U), PUFA 3

(product number 47085-U) and PUFA 1 (product number

47033-U). Chromatograms were integrated using the Varian

Galaxie Chromatography Data System, version 1.9.3.2 (Agi-

lent Technologies, Colorado Springs, CO, USA).

Feed samples and muscle tissue samples were homogenized

using a Polytron homogenizer in 10 mL ultrapure (Milli-Q,

EMD Millipore, Billerica, MA, USA) water. The homoge-

nizer was washed three times between samples using ultra-

pure water. An aliquot of the homogenate (0.5 mL) was

hydrolysed with 0.5 mL HCl/phenol (1% by weight) at

110 °C in microreaction vials for 24 h.

Total amino acids were derivatized using an EZ:faastTM

GC-FID Amino Acid Analysis Kit (Phenomenex). Following

hydrolysis and derivatization, samples were run on a Varian

3800 GC-FID with a column length of 10 m and a diameter

of 0.25 mm (ZB – AAA Zebron Amino Acid; Phenomenex).

The injector maintained a constant temperature of 250 °C

and used 2.0 lL of sample with a 1 : 15 split. The column

temperature began at 110 °C, was ramped to 320 °C at a rate

of 32 °C min�1 and held for 2 min to ensure elution of all

amino acids. The carrier gas, helium, flowed at a constant

rate of 1.5 mL min�1. The detector temperature stayed con-

stant at 320 °C. Peaks were integrated using the Varian Gal-

axie Chromatography Data System, version 1.9.3.2 (Walnut

Creek, CA, USA), to obtain a quantitative amino acid pro-

file. Taurine and arginine are not determined using the EZ:

faastTM method. Peak areas were identified and quantified in

comparison with an internal standard and a four-level cali-

bration curve (level 1: 50 nmols mL�1, level 2: 100 nmols

mL�1, level 3: 150 nmol mL�1 and level 4: 200 nmol ml�1).

Standard solutions were supplied with the EZ:faastTM kit.

Muscle tissue samples were dried at 80 °C for 24 h, finely

ground, fumed in a desiccator containing concentrated HCl

for another 24 h, dried for another 24 h at 80 °C and then

run on Perkin Elmer analyzer (Series II; CHNSO Analyzer

2400, Waltham, MA, USA).

Fish were sampled for sensory analysis 2 weeks after final

sampling for lipid analyses (week 18) from the 0 CM and

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24 CM treatments. Three fish from each dietary group

were killed by a blow to the head, gutted and chilled on

ice. Fish were then filleted and portioned into 3 cm 9 3 cm

squares, placed into sampling cups and covered with lids.

Raw samples were carried to an isolated, air-freshened

room with standardized light at the Ocean Sciences Centre.

The room was prepared according to standards outlined by

Carpenter et al. (2000). An untrained panel (n = 24) com-

posed of volunteers performed a triangle test, a hedonic

test and a quantitative descriptive analysis (QDA) to profile

salmon treatments in terms of smell, appearance and tex-

ture. Panelists were given specific instructions about how to

evaluate the samples for appearance, odour and texture

(Carpenter et al. 2000). The triangle test required panelists

to determine which one of the three samples seemed differ-

ent in terms of appearance, texture and odour (two 0%

CM samples and one 24% CM sample). The hedonic test

required panelists to score on a two-anchored (e.g. firm

and soft) linear scale (1–7) for each of the appearance,

odour and texture (FAO, 1999). The QDA asked panelists

to rate the intensity on a scale of 1–7 (1 = no intensity;

7 = distinct intensity) for a number of descriptions (e.g.

brightness, orange intensity, surface moistness). Both QDA

tests were objective and had nothing to do with the likes or

dislikes of each panelist.

Growth performance data were analysed using nonlinear

regression (quadratic) (Shearer 2000). A two-level nested

ANOVA was used to confirm the results of the nonlinear

regression and to differentiate treatments with a post hoc

analysis (Tukey’s HSD) when significant differences

occurred (P < 0.05) and was performed using the general

linear model (Minitab 16 Statistical Software, State Col-

lege, PA, USA). The two-level nested ANOVA model tested

the effect of diet on the response variable and nested fish

individuals within tanks to negate variability among tanks

and individuals, and also testing for tank effects. Tissue

composition data were analysed using this model. For

analysis of growth data that depend on comparison to an

initial measurement and thus must be pooled per tank

(i.e. weight gain, specific growth rate, feed intake, feed

conversion ratio), a one-way ANOVA was performed to test

the effect of diet. For each model tested, the residuals

were examined to evaluate the appropriateness of the

model; therefore, normality, homogeneity and indepen-

dence of residuals were considered. If a P-value was

close to 0.05 and residuals were not normal, a P-value

randomization was conducted >5000 times to test the data

empirically.

In addition, PRIMER (Plymouth Routines in Multivari-

ate Ecological Research; PRIMER-E Ltd, Version 6.1.15,

Ivybridge, UK) was used to analyse selected fatty acid and

amino data, using similarity of percentages analysis (SIM-

PER), analysis of similarities (ANOSIM), principal components

analysis (PCA) and multidimensional scaling (MDS) to

define similarities and differences among tissue and dietary

fatty acid data. Fatty acids and amino acids that accounted

for > 1% of total fatty acids were included in the analyses.

Multidimensional scaling, SIMPER and ANOSIM are multivari-

ate analyses that use a resemblance matrix, and the latter

carries out an approximate analogue of ANOVA. In both

cases, nonparametric Bray–Curtis similarity was chosen.

Multivariate analyses (MDS and PCA) that compared fatty

acid or amino acid profiles between species were focused

on 14% CM (trout) and 16% CM (salmon) diets instead of

the highest CM levels (21% CM and 24% CM) because

the diets were more comparable in terms of CM and FM

levels. The data used in the PCA that included total lipids,

fatty acids, amino acids and growth performance variables

were normalized prior to analysis in order to standardize

the data measured on different scales.

Solvent-extracted CM contained 5.53% ww�1 total lipid,

with similar levels of triacylglycerol (29.4%) and phospho-

lipid (32.3%) (Table 1). 18:2n-6 (27.2%) and 18:3n-3

(26.7%) were the most abundant fatty acids in the CM

lipid fraction. Glutamine was the most abundant amino

acid. The remaining amino acids in CM contributed <10%

each to the total amino acids. Crude protein and gross

energy digestibility of CM by rainbow trout and Atlantic

salmon were previously determined (Fraser & Anderson

2012). Antinutritional factors present were glucosinolates,

fibre, phytate and non-starch polysaccharides (Table 1).

The experimental diets included 70 g kg�1, 140 g kg�1 and

210 g kg�1 CM, with 0 g kg�1 CM as a control (Table 2).

Protein did not differ among diets, but the amount of lipid

varied significantly (142–195 g kg�1) (Supporting Informa-

tion). Triacylglycerol levels were higher in the 0% CM and

14% CM diets than 7% CM and 21% CM. Sterols

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increased significantly with the addition of CM in the diets.

Several fatty acids were significantly different among diets,

which was either the result of CM addition or the differ-

ence in the level of FM in the diets, notably 18:2n-6, 18:3n-

3, 20:1n-9. Several amino acids in the diets different signifi-

cantly; however, alanine and proline decreased significantly

with greater CM inclusion.

The experimental diets included 80 g kg�1, 160 g kg�1 and

240 g kg�1 CM, with 0 g kg�1 CM as a control (Table 1).

Protein did not differ among diets, but the amount of lipid

varied significantly (17.6–25.1%) (Supporting Information).

Triacylglycerol and sterols increased significantly with the

addition of CM in the diets. Several fatty acids were signifi-

cantly different among diets; however, the level of fatty

acid did not necessarily depend on the level of CM inclu-

sion. Leucine and proline decreased significantly with

greater CM inclusion, while lysine and tyrosine increased

significantly with greater CM inclusion.

In all cases, the growth data appropriately fit the quadratic

regression model and the regression results were confirmed

using ANOVA (Supporting Information). The final weight of

rainbow trout varied significantly depending on inclusion of

CM in the diet, with trout on the 21% CM diet having the

lowest final weight and gaining the least amount of weight

after 12 weeks (Table 3). The growth rate followed a similar

pattern; where trout fed a diet without CM grew the fastest,

trout fed the 21% CM diet grew the slowest. The feed con-

version ratio was highest for trout fed the 21% CM diet and

lowest for trout fed the 0% CM and 7% CM diets. The pro-

tein efficiency ratio was lowest in trout fed the 21% CM

diet. There were no significant differences among tanks per

treatment for any growth parameter.

Salmon that were fed any diet containing CM had a sig-

nificantly lower final weight and gained less weight than

the salmon that were fed the 0% CM diet after 16 weeks

of growth (Table 3). Salmon fed any CM diet also con-

sumed significantly less feed throughout the experiment

compared with salmon fed the diet without CM; however,

the feed conversion ratio was not different among treat-

ments. The visceral somatic index was higher in salmon fed

CM diets than the control.

Table 1 Characterization of solvent-extracted camelina meal1

Proximate composition

Solvent-extracted

camelina meal, as fed

Moisture (g kg�1)2 75.0

Crude protein (g kg�1)2 390

Gross energy (kcal kg�1)2 4320

Total lipid (% ww�1) 5.53 � 0.4

Triacylglycerol (% total lipid) 29.4 � 8.2

Phospholipid (% total lipid) 32.3 � 2.2

Fatty acids (weight %)

18:1x9 16.1 � 0.3

18:2x6 27.2 � 0.8

18:3x3 26.7 � 1.1

20:1x9 7.8 � 0.6

20:2x6 1.4 � 0.2

22:1x9 1.1 � 0.1

Amino acids (g/16 g N)

Alanine 4.1 � 0.3

Sarcosine 1.6 � 0.1

Glycine 5.4 � 0.3

Valine 5.9 � 0.5

Leucine 6.4 � 0.3

Isoleucine 4.2 � 0.3

Threonine 4.0 � 0.2

Serine 4.4 � 0.2

Proline 7.0 � 0.5

Thioproline 0.4 � 0.1

Aspartic acid 8.5 � 0.9

Methionine 1.6 � 0.2

Hydroxyproline 1.3 � 0.2

Glutamic acid 9.6 � 0.8

Phenylalanine 4.0 � 0.3

Glutamine 29.1 � 2.3

Lysine 2.9 � 0.3

Histidine 3.8 � 0.1

Tyrosine 2.1 � 0.1

Total amino acid (%) 37.8 � 1.2

Apparent digestibility coefficients3

Crude protein digestibility (%) 83.9 (RT) 85.8 (AS)

Gross energy digestibility (%) 57.4 (RT) 62.3 (AS)

Antinutritional factors

Glucosinolates4 (l mol g�1) 38.1

Fibre (acid detergent)5 (g kg�1) 184

Fibre (neutral detergent)5 (g kg�1) 342

Phytate6 (g kg�1) 6.50

Non-starch polysaccharides7 (g kg�1) 251

1 Summarized from Hixson et al. (2014a,b).2 Ye (2014).3 Fraser & Anderson (2012) (RT = rainbow trout; AS = Atlantic sal-

mon).4 Analysis completed by Agriculture and Agri-food Canada – Sas-

katoon Research Centre (Saskatoon, SK, Canada).5 Analysis completed by Nova Scotia Department of Agriculture

(Truro, NS, Canada).6 Analysis completed by MCN Bioproducts Inc. (Saskatoon, SK,

Canada).7 Analysis completed by University of Manitoba (Winnepeg, MB,

Canada).

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


In rainbow trout, there was no significant difference

depending on diet for measures of total hydrogen, total

nitrogen, carbon to nitrogen ratios, nitrogen intake or

nitrogen gain (Table 4). Trout fed 21% CM retained

more nitrogen in the muscle than trout fed 0% CM

and 7% CM. Total carbon in the muscle tissue was

significantly different between trout fed 7% CM and

21% CM, but did not differ among the remaining

treatments.

In Atlantic salmon, there were no significant differ-

ences in carbon, nitrogen, hydrogen and the carbon to

nitrogen ratio (Table 4). Nitrogen intake was signifi-

cantly greater in salmon fed the 0% CM diet, but did

not differ among salmon fed diets containing CM. Sal-

mon fed the 16% CM diet retained significantly more

nitrogen than salmon fed the 0% CM diet; however,

there were no significant differences among the remain-

ing groups.

Rainbow trout fed the 21% CM diet had significantly

higher lipid levels in the muscle tissue than trout fed the

0% CM diet (Table 5). Trout fed the 0% CM diet stored

less neutral lipid and more polar lipid in their muscle than

trout fed any diet containing CM. Sterols were lower in the

muscle of trout fed the 0% CM diet compared with trout

fed any diet containing CM. Phospholipid and triacylglyc-

erol levels in the muscle tissue were not different among

treatments. Very few fatty acids in the muscle differed

depending on the treatment (Table 5). Levels of 18:3n-3

were significantly lower in trout fed the 0% CM diet com-

pared with trout fed any diet containing CM and did not

differ among trout fed varying levels of CM. Total n-6

fatty acids were significantly higher in trout fed 0% CM

than trout fed any diet with CM, while the CM groups did

not differ among themselves.

Salmon fed 0% CM had significantly lower levels of lipid

in the muscle tissue than salmon fed any diet containing

CM (Table 5). Neutral lipid levels were significantly higher

Table 2 Formulation and proximate composition of control and experimental diets for rainbow trout and Atlantic salmon

Ingredient (g kg�1)

Rainbow trout Atlantic salmon

0% CM 7% CM 14% CM 21% CM 0% CM 8% CM 16% CM 24% CM

Fish meal 335.0 315.0 290.0 352.0 349.0 324.0 299.0 274.0

Camelina meal – 70.0 140.0 210.0 – 80.0 160.0 240.0

Fish oil 175.0 183.0 194.0 193.0 140.0 157.0 173.0 189.0

Soybean meal 100.0 100.0 100.0 100.0 –Empyreal 80.0 80.0 80.0 80.0 50.0 50.0 50.0 50.0

Wheat gluten meal – – – – 150.0 150.0 150.0 150.0

Whey – – – – 50.0 50.0 50.0 50.0

Wheat middlings 168.0 109.0 94.0 – 224.0 153.0 82.0 10.0

Feather meal 50.0 50.0 50.0 50.0 – – – –Poultry by-product meal 50.0 50.0 50.0 50.0 – – – –Pregelatinized starch – – – – 25.0 25.0 25.0 25.0

Lignosol 25.0 25.0 25.0 25.0 – – – –Salt (de-iodized) 3.0 3.0 3.0 3.0 – – – –D/L methionine 2.0 2.0 2.0 2.0 1.7 1.7 1.7 1.7

Vitamin/mineral premix1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0

Pigment/antioxidant premix2 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5

Choline chloride 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0

Lysine-HCl 3.0 3.0 3.0 3.0 – – – –Proximate composition analysed (as fed, g kg�1) (n = 3)

Dry matter 891.0 893.0 894.0 908.0 914.0 917.0 932.0 916.0

Ash 32.0 29.0 31.0 30.0 29.0 25.0 31.0 27.0

Crude protein 444.0 463.0 449.0 456.0 413.0 411.0 424.0 410.0

Lipid 142.0 140.0 180.0 195.0 176.0 195.0 227.0 251.0

1 Vitamin/mineral premix contains per kg: Zinc 77.5 mg, Manganese 125 mg, Iron 84 mg, Copper 2.5 mg, Iodine 7.5 mg, Vitamin A

5000 IU, Vitamin D 4000 IU, Vitamin K 2 mg, Vitamin B12 4 lg, Thiamine 8 mg, Riboflavin 18 mg, Pantothenic acid 40 mg, Niacin

100 mg, Folic acid 4 mg, Biotin 0.6 mg, Pyridoxine 15 mg, Inositol 100 mg, Ethoxyquin 42 mg, Wheat shorts 1372 mg.2 Antioxidant/pigment premix contains per kg: Selenium 0.220 mg, Vitamin E 250 IU, Vitamin C 200 mg, Astaxanthin 60 mg, Wheat

shorts 1988 mg.

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


Table

3Growth

perform

ance

ofsalm

onidsfeddiets

containingcamelinameal1;rainbow

trout2

werefedexperim

entaldiets

for12weeks;

andAtlanticsalm

on3werefeddiets

for

16weeks

Growth

parameter

Rainbow

trout

Atlanticsalm

on

0%

CM

7%

CM

14%

CM

21%

CM

r2P-value

0%

CM

8%

CM

16%

CM

24%

CM

r2P-value

Initialweight

43.4

�12

45.3

�10

44.1

�10

45.9

�10

0.01

0.839

230�

41

246�

62

250�

42

241�

49

0.03

0.637

Finalweight

168�

8.1

a164�

19ab

158�

5.4

b137�

8.3

c0.09

0.037

691�

153a

576�

152b

560�

129b

565�

117b

0.11

0.001

Weightgain

4125�

3.5

a119�

20ab

114�

6.7

ab

90.6

�4.7

b0.65

0.009

471�

39a

329�

72b

309�

45b

327�

17b

0.72

0.003

Initiallength

15.4

�1.2

15.8

�1.3

15.6

�1.2

15.7

�1.1

0.01

0.736

26.2

�2.4

26.9

�1.9

27.5

�1.5

26.8

�1.6

0.05

0.424

Finallength

23.3

�1.4

a22.7

�1.9

ab

22.5

�1.9

ab

21.3

�2.4

b0.12

0.009

35.0

�4.1

33.8

�2.8

33.4

�2.6

33.3

�2.9

0.04

0.081

CF5

1.32�

0.08

1.36�

0.08

1.36�

0.08

1.36�

0.09

0.03

0.274

1.53�

0.1

1.46�

0.1

1.48�

0.1

1.54�

0.5

0.01

0.106

VSI

612.4

�1.2

13.1

�1.7

13.4

�1.3

13.5

�1.8

0.08

0.058

9.8

�1.1

a10.8

�1.0

b11.2

�1.2

b11.4

�0.9

b0.25

0.001

AFI

7100.9

�4.5

107.0

�5.2

100.8

�6.1

104.9

�6.8

0.14

0.498

515�

7.6

a420�

57b

384�

33b

391�

20b

0.78

0.001

FCR8

0.86�

0.05a

0.93�

0.07a

1.04�

0.02ab

1.12�

0.06b

0.52

0.035

1.01�

0.1

1.20�

0.2

1.16�

0.1

1.10�

0.1

0.26

0.299

PER9

2.6

�0.2

a2.4

�0.2

ab

2.1

�0.01ab

1.9

�0.3

b0.69

0.005

2.22�

0.2

1.90�

0.3

1.89�

0.2

2.04�

0.05

0.34

0.001

1P-valueandr2

determ

inedbyquadraticregression.Meanswithdifferentsuperscripts

indicate

significantdifferencesamongtreatm

ents

determ

inedbyTukey’sHSD

(P>0.05).

2Rainbow

troutva

luesare

mean�

SD;initialmeasurements

(n=9),finalmeasurements

(n=18).

3Atlanticsalm

onva

luesare

mean�

SD;initialmeasurements

(n=9),finalmeasurements

(0%

CM

=48;8%

CM

=66,16%

CM

=69,24%

CM

=50).Meanswithdifferentsuper-

scripts

indicate

significantdifferencesamongtreatm

ents

(P>0.05).

4Weightgain

(gfish

�1)=finalweight�

initialweight(calculatedbytankmeans,

n=3).

5Conditionfactor=bodymass/length

3(calculatedbyindividualfish).

6Visceralsomaticindex(%

)=100*(viscera

mass/bodymass).

7Apparentfeedintake(g

fish

�1)=(feedco

nsumed,g)/(numberoffish

pertank)(calculatedbytankmeans,

n=3).

8Fe

edco

nve

rsionratio(g

fish

�1)=(feedintake,gfish

�1)/(w

eightgain,gfish

�1)(calculatedbytankmeans,

n=3).

9Protein

effi

ciency

ratio(g

gain

pergprotein

intake)=(w

eightgain,gfish

�1)/(protein

intake,gfish

�1).

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


in salmon fed the 8% CM and 16% CM diets than in sal-

mon fed the 0% CM and 24% CM diets, while polar lipid

levels were significantly lower in salmon fed the 0% CM

diet than any other treatment. Few fatty acids in the mus-

cle differed depending on the treatment (Table 5). Levels of

18:3n-3 increased significantly in the muscle tissue with

increased levels of dietary CM. 20:5n-3 was significantly

higher in salmon fed the 24% CM diet than salmon fed

both 0% CM and 8% CM diets, while 20:5n-3 in salmon

fed 16% CM was not different with any group. 22:1n-9

was significantly lower in salmon fed 0% CM than salmon

fed any diet containing CM. 22:6n-3 followed the same pat-

tern.

In rainbow trout, individual amino acids did not differ

depending on the diet consumed (Table 6). In salmon, sev-

eral individual amino acids different significantly among

groups, including the following: threonine, aspartic acid,

glutamic acid, phenylalanine, lysine and tyrosine (Table 6).

However, amounts of these amino acids did not necessarily

increase or decrease depending on the level of CM in the

diet, except for lysine which decreased with increased levels

of CM and glutamic acid which increased with dietary

CM.

ANOSIM and SIMPER were conducted to evaluate differences

in muscle tissue fatty acid composition of rainbow trout

and salmon fed diets with varying levels of CM (Support-

ing Information). Trout fed the 0% CM diet had signifi-

cantly different fatty acid profiles to trout that were fed the

14% CM, 21% CM and 7% CM diets (in order of dissimi-

larity), while trout fed the 7% CM had the most similar

fatty acid profiles to trout fed either the 14% CM or the

21% CM diets. 22:6n-3 and 22:1n-9 were mainly responsi-

ble for the differences between fatty acid profiles of trout

fed different diets. Atlantic salmon fed the 0% CM diet

were significantly different to salmon fed the 24% CM,

16% CM and 8% CM diets (in order of dissimilarity).

22:6n-3 was the major contributing fatty acid to the differ-

ences in fatty acid profiles from salmon fed the 0% CM

diet compared to the CM diets.

The MDS plot illustrated distinct separation of fatty acid

profiles clustered according to species and treatment

(Fig. 1). Fatty acid profiles from the muscle tissue of

Atlantic salmon clustered separately from those of rainbowTable

4Carbon,nitrogen

andhydrogen

concentrations(m

gg�1)in

muscle

tissueofrainbow

trout(12weeks)

andAtlanticsalm

on(16weeks)

fedcamelinamealdiets1

Rainbow

trout

Atlanticsalm

on

0%

CM

7%

CM

14%

CM

21%

CM

P-value

0%

CM

8%

CM

16%

CM

24%

CM

P-value

Carbon(m

gg�1)

583�

22ab

568�

13a

571�

14ab

598�

15b

0.029

532�

18

552�

27

548�

29

561�

26

0.536

Nitrogen(m

gg�1)

124�

6.3

125�

7.2

127�

8.3

121�

7.7

0.517

108�

10

93.3

�14

100�

16

94.7

�13

0.197

Protein

(mgg�1)

775�

39

781�

45

796�

52

756�

45

0.517

677�

64

583�

88

626�

97

592�

84

0.197

Hyd

rogen(m

gg�1)

82.5

�2.1

82.8

�2.1

85.6

�2.1

83.7

�4.8

0.381

80.4

�3.4

84.0

�5.4

82.4

�5.6

83.4

�4.1

0.687

Carbon:Nitrogen

4.71�

0.4

4.57�

0.4

4.51�

0.4

4.97�

0.4

0.202

4.97�

0.6

6.1

�1.1

5.6

�1.2

6.0

�1.1

0.193

Nitrogenintake2

3.50�

0.4

3.84�

0.2

3.86�

0.01

3.87�

0.1

0.085

34.0

�0.4

a27.6

�3.2

b26.0

�1.9

b25.7

�1.1

b<0.001

Nitrogengain

31.09�

0.4

1.16�

0.7

1.42�

0.5

1.56�

0.4

0.333

1.21�

0.3

0.89�

0.6

1.15�

0.5

1.0

�0.4

0.431

Nitrogenretention4

32.0

�5.6

a30.0

�6.7

a36.7

�5.1

ab

40.3

�3.8

b0.015

35.8

�7.6

a37.0

�7.3

ab

44.7

�7.0

b39.0

�5.3

ab

0.046

1Valuesare

mean(n

=9)�

SD.Meanswithdifferentsuperscripts

indicate

significantdifferencesamongdiets

foreach

species.

2Nitrogenintake(g

fish

�1)=feedintake*nitrogenin

feed.

3Nitrogengain

(gfish

�1)=(finalbodynitrogen–initialbodynitrogen)/(w

eightgain).

4Nitrogenretention(%

)=100*(nitrogengain/nitrogenintake).

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


Table

5Lipid

class

andfattyacidcompositionin

themuscle

tissueofrainbow

troutandAtlanticsalm

onsm

olts1

Composition

Rainbow

trout

Atlanticsalm

on

0%

CM

7%

CM

14%

CM

21%

CM

P-value

0%

CM

8%

CM

16%

CM

24%

CM

P-value

%ww

�1

Totallipid

11.5

�5.7

a14.5

�2.2

ab

17.9

�3.9

ab

22.4

�3.9

b0.038

14.0

�2.0

a26.9

�6.7

b26.7

�17b

26.5

�8.5

b0.001

Neutrallipid

4.7

�3.6

a12.3

�2.0

b14.8

�3.3

b18.9

�4.1

b0.003

12.6

�2.3

a22.6

�8.1

b21.3

�11b

15.6

�8.0

a0.001

Polarlipid

6.7

�2.3

a2.2

�0.3

b3.0

�1.4

b3.5

�0.6

b0.011

1.5

�0.3

a4.1

�1.8

b10.7

�6.1

b11.0

�3.1

b0.001

%Totallipid

Triacylglycerol

74.9

�6.3

75.9

�2.5

75.9

�6.9

75.9

�11

0.993

88.7

�12a

71.3

�10b

70.2

�12b

69.4

�11c

0.001

Freefattyacid

2.2

�0.5

a3.7

�0.7

b3.2

�0.6

b3.1

�1.0

b0.015

2.0

�1.0

3.5

�1.0

2.0

�1.0

5.0

�1.7

0.088

Sterol

1.6

�0.4

a4.3

�0.7

b3.3

�1.0

b3.5

�1.2

b0.001

0.5

�0.2

3.4

�1.2

3.0

�1.0

1.7

�0.3

0.198

Phospholipid

17.2

�6.6

12.6

�1.5

14.6

�6.2

19.3

�9.0

0.318

4.9

�1.0

a11.4

�3.8

b10.0

�2.9

b8.6

�3.8

ab

0.034

Fattyacids2

14:0

4.7

�0.4

4.8

�0.2

5.0

�0.5

4.8

�0.5

0.492

5.2

�0.3

5.1

�0.3

5.2

�0.2

5.3

�0.2

0.248

16:0

15.5

�0.5

15.8

�0.5

15.5

�0.4

15.1

�0.6

0.366

15.7

�0.6

15.5

�0.6

15.5

�0.3

15.4

�0.5

0.664

16:1n-7

7.1

�0.3

7.1

�0.2

7.4

�0.3

7.1

�0.5

0.270

7.5

�0.5

7.8

�0.2

7.8

�0.2

8.0

�0.1

0.784

18:0

3.2

�0.1

3.2

�0.1

3.1

�0.1

3.0

�0.1

0.127

3.4

�0.2

3.3

�0.1

3.3

�0.1

3.3

�0.1

0.120

18:1n-9

13.0

�0.3

13.2

�0.5

13.6

�0.4

13.5

�0.2

0.549

14.2

�1.2

13.7

�0.8

13.8

�0.9

13.6

�1.1

0.591

18:2n-6

5.7

�0.2

5.4

�0.3

5.3

�0.5

4.8

�0.9

0.157

5.6

�0.4

5.4

�0.2

5.4

�0.3

5.3

�0.3

0.204

18:3n-3

1.2

�0.06a

1.6

�0.1

b1.5

�0.1

b1.6

�0.2

b0.010

0.9

�0.1

a1.4

�0.1

b1.7

�0.1

c2.0

�0.1

d0.001

20:1n-9

3.0

�0.2

3.1

�0.1

3.0

�0.2

3.1

�0.1

0.664

2.8

�0.3

3.1

�0.1

3.0

�0.2

2.7

�0.1

0.999

20:4n-6

0.9

�0.05

0.8

�0.02

0.8

�0.03

0.9

�0.09

0.532

0.8

�0.1

0.8

�0.03

0.8

�0.02

0.8

�0.03

0.997

20:5n-3

10.2

�0.4

10.3

�0.3

10.5

�0.3

10.6

�0.8

0.582

9.8

�0.7

a9.8

�0.4

a9.3

�0.5

ab

9.0

�0.5

b0.029

22:1n-9

2.8

�0.2

2.8

�0.2

2.6

�0.1

2.7

�0.2

0.615

0.3

�0.05a

0.5

�0.02b

0.5

�0.04b

0.5

�0.04b

0.009

22:5n-3

3.0

�0.1

3.1

�0.1

3.1

�0.2

3.0

�0.3

0.460

3.8

�0.1

3.6

�0.3

3.6

�0.1

3.6

�0.2

0.108

22:6n-3

15.6

�1.2

14.5

�0.6

14.0

�1.1

15.3

�1.1

0.131

12.4

�1.0

a10.9

�0.7

b10.8

�0.5

b10.4

�0.4

b0.001

∑SFA3

23.7

�1.2

24.7

�0.8

24.3

�0.6

23.6

�0.9

0.375

25.2

�0.6

24.9

�0.8

25.1

�0.5

25.2

�0.8

0.757

∑MUFA

431.1

�1.7

31.6

�0.6

31.9

�0.7

31.9

�0.7

0.572

33.6

�0.7

a34.7

�0.6

b34.0

�0.7

ab

33.3

�0.8

a0.006

∑PUFA

544.5

�1.2

43.2

�0.8

43.2

�1.1

43.8

�1.1

0.257

40.5

�0.8

39.7

�0.6

40.2

�0.6

40.7

�0.5

0.082

∑n-3

32.9

�1.4

32.2

�0.7

32.1

�1.4

33.4

�1.0

0.462

28.1

�0.8

28.1

�0.6

28.7

�0.7

29.2

�0.6

0.075

∑n-6

7.7

�0.3

a6.4

�0.2

b6.3

�0.5

b5.8

�1.0

b0.004

7.6

�0.6

7.4

�0.3

7.4

�0.3

7.4

�0.4

0.413

1Valuesare

mean(n

=9)�

SD.Meanswithdifferentsuperscripts

indicate

significantdifferencesamongdiets

foreach

species.

2Data

exp

ressedasareapercentageoffattyacidmethyl

ester.

3Sa

turatedfattyacid.

4Monounsaturatedfattyacid.

5Polyunsaturatedfattyacid.

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


trout, and fish fed 0% CM diets clustered separately from

fish that were fed 14% or 16% CM diets. A cluster analysis

confirmed the groups observed in the MDS plot, with

Atlantic salmon and rainbow trout separated from one

common branch, which further separated into treatment

groups from each species branch (Supporting Information).

The partition lines in the MDS plot (Fig. 1) were based on

the groupings revealed by the cluster analysis. Two individ-

uals did not cluster with the others; however, they were not

considered outliers in the original data set, as determined

by Dixon’s outlier test for rainbow trout (P = 0.117) and

Atlantic salmon (P = 0.215).

ANOSIM and SIMPER were conducted to evaluate differences

in muscle tissue amino acid composition of rainbow trout

and salmon fed diets with varying levels of CM (Support-

ing Information). The amino acid profile of rainbow trout

fed 7% CM and 14% CM were the most dissimilar,

although the difference was not significant, while trout fed

0% CM and 21% CM had the most similar amino acid

profiles. The minimal dissimilarity that was observed was

due to lysine, which contributed 19.6–21.1% to the overall

dissimilarity between amino acid profiles. The amino acid

profiles of Atlantic salmon fed the 0% CM and 8% CM

diets were significantly different according to ANOSIM, and

alanine and valine were most responsible for the differences

in amino acid profiles between groups.

A PCA based on amino acid profiles from the muscle tis-

sue of rainbow trout and Atlantic salmon fed either 0%

CM, 14% CM (rainbow trout) or 16% CM (Atlantic sal-

mon) illustrated that amino acid profiles from trout and

salmon are different by occupying the negative (trout) and

positive (salmon) regions of PC1 (Fig. 2). There was no

separation between treatments within a species however,

and PC1 and PC2 explained only approximately half of the

variation in the data (53.5%). Amino acid vectors were

selected based on a correlation > 0.6.

Table 6 Amino acid composition in the muscle tissue of rainbow trout and Atlantic salmon smolts1,2

Rainbow trout Atlantic salmon

0% CM 7% CM 14% CM 21% CM P-value 0% CM 7% CM 16% CM 24% CM P-value

Alanine 7.2 � 0.7 7.1 � 0.7 7.5 � 1.1 7.2 � 0.4 0.887 10.5 � 2.9 9.5 � 2.4 12.1 � 2.2 11.6 � 2.9 0.676

Valine 7.5 � 1.3 7.7 � 1.4 7.1 � 1.1 7.0 � 0.9 0.650 8.4 � 1.5 8.6 � 1.4 10.6 � 4.9 10.4 � 3.8 0.338

Leucine 10.0 � 0.6 9.9 � 0.4 10.2 � 0.8 10.0 � 0.5 0.885 11.3 � 1.2 11.2 � 0.9 12.6 � 1.2 13.0 � 1.9 0.156

Threonine 6.1 � 0.7 5.8 � 0.8 6.0 � 1.0 6.1 � 0.5 0.938 5.7 � 1.0a 7.0 � 1.1b 7.0 � 0.5b 7.2 � 0.8b 0.039

Aspartic acid 13.4 � 1.7 12.8 � 1.7 13.0 � 1.5 13.4 � 1.2 0.947 11.4 � 2.1a 10.1 � 2.3b 11.0 � 2.0a 13.4 � 1.0c 0.008

Methionine 3.9 � 0.3 4.0 � 0.3 4.0 � 0.8 4.1 � 0.4 0.885 2.9 � 0.4 3.0 � 0.4 3.0 � 0.5 2.9 � 0.6 0.996

Glutamic acid 14.2 � 0.9 14.2 � 1.1 13.2 � 0.2 13.4 � 1.2 0.391 14.5 � 1.8a 14.8 � 0.8a 17.6 � 0.3b 18.4 � 2.8b 0.040

Phenylalanine 5.3 � 0.4 5.2 � 0.2 5.3 � 0.3 5.3 � 0.2 0.995 7.6 � 0.8ab 6.2 � 0.8a 9.0 � 1.8b 8.5 � 0.9b 0.019

Lysine 15.2 � 2.6 16.5 � 3.8 15.4 � 4.1 14.6 � 3.4 0.931 13.0 � 0.9a 11.7 � 1.0b 9.8 � 0.9bc 9.4 � 1.0c 0.023

Histidine 4.0 � 1.9 3.9 � 1.9 4.6 � 0.3 4.8 � 0.2 0.627 1.3 � 0.5 1.9 � 0.5 1.7 � 0.6 1.0 � 0.4 0.087

Tyrosine 4.4 � 0.2 4.6 � 0.4 4.6 � 0.2 4.6 � 0.2 0.876 7.8 � 0.9a 7.2 � 0.8ab 7.0 � 0.9b 7.4 � 0.6ab 0.041

∑ Total 120 � 3.7 119 � 4.2 118 � 1.0 120 � 2.0 0.635 125 � 7.1 123 � 5.6 128 � 10 129 � 6.0 0.293

∑ Essential 57.7 � 2.1 58.1 � 3.7 55.5 � 6.1 57.0 � 3.1 0.816 56.7 � 6.5 64.7 � 5.6 61.2 � 6.0 59.0 � 6.2 0.055

∑ Non

-essential

62.9 � 4.2 60.7 � 6.6 62.7 � 7.0 62.6 � 5.0 0.952 68.6 � 9.7ab 57.9 � 5.4a 67.1 � 8.7ab 69.8 � 10b 0.026

1 Values are mean (n = 9) � SD. Means with different superscripts indicate significant differences among diets for each species.2 Data expressed as g 16 g�1 nitrogen.

Figure 1 Multidimensional scaling plot with fatty acid profiles

from muscle tissue of rainbow trout and Atlantic salmon fed diets

with camelina meal (14CM for rainbow trout and 16CM for

Atlantic salmon) and without camelina meal (0CM).

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


A PCA was conducted to relate growth performance

parameters, fatty acid, amino acid, total lipid and lipid

class levels in muscle tissue for both rainbow trout and

Atlantic salmon. In the rainbow trout PCA (Fig. 3a), sev-

eral fatty acids (e.g. 18:3n-3, 22:5n-3, 18:1n-7, 16:1n-7) and

amino acids (e.g. methionine, serine, threonine, glycine,

proline, aspartic acid) grouped together. Lipid classes also

associated with fatty acids, such as sterols, phospholipids,

22:6n-3 and 20:4n-6. However, there were also visible asso-

ciations with both amino acids and fatty acids, which were

in close proximity to each other, such as total lipids and

neutral lipids with leucine and 20:1n-9 with glutamic acid.

The first two components (PC1 + PC2) explained 90.7% of

the variation.

The Atlantic salmon PCA (Fig. 3b) showed different

associations between fatty acids and amino acids than was

observed in the rainbow trout PCA. Several relevant

groupings were observed, such as glycine, tyrosine, leucine

and 16:1n-7; total lipid, phospholipid, lysine and 22:6n-3;

and histidine, 20:1n-9 and neutral lipid. The first two com-

ponents (PC1 + PC2) explained 89.6% of the variation.

Panelists could not distinguish the fillet sample from the

24% CM treatment compared to two fillet samples from

the 0% CM treatment. There were no significant differ-

ences between fillets from the 0% CM treatment and 24%

CM treatment for the parameters tested in the QDA:

brightness (t-stat = 0.380; P = 0.71), orange intensity (t-

stat = 1.33; P = 0.189), surface moistness (t-stat = 0.390;

P = 0.493), firmness (t-stat = �0.630; P = 0.530), marine

odour (t-stat = �0.470; P = 0.638), vegetable odour (t-

stat = 0.200; P = 0.841) and rancid odour (t-stat = 0.690,

P = 0.496). There was no significant difference in appear-

ance (t-stat = 1.28; P = 0.206), odour (t-stat = 0.510;

P = 0.612) and texture (t-stat = 1.26; P = 0.215).

FM is no longer the main protein source in aquaculture

feeds. Instead, a variety of sustainable and economical

ingredients are used to increase the protein content and

balance the amino acid profile, while FM is typically pro-

vided at minimum levels. In the present study, CM was

investigated as a potential protein source to include in diets

for farmed salmonids. Including CM in diets for salmonids

impacted their growth after at least 12 weeks. Rainbow

trout generally performed better than Atlantic salmon

when fed diets including CM and could tolerate up to 14%

CM included in the diet without adversely affecting the

growth performance, while salmon was impacted with only

8% CM inclusion. The difference in the level of tolerance

between species could have been a result of species, age

class, environment and duration of the experiments. Even

within the same species fed a fixed dietary formulation, fish

size and feeding regime can affect growth and digestibility

(Jobling 1983). However, despite the differences between

the two experiments, both salmonid species could not toler-

ate high levels (>14% inclusion) of CM in their diets.

Increased inclusion level of plant proteins in diets for sal-

monids typically results in decreased growth rates, as

reviewed by Collins et al. (2013) in a meta-analysis of

growth data from literature on salmonids fed diets includ-

ing six different plant proteins. In this study, increased CM

inclusion was associated with a decrease in feed consump-

tion and weight gain in salmon, and decreased weight gain

and increased feed conversion ratio in trout, indicating low

utilization of nutrients in both species. The effect on

growth performance was likely due to the antinutritional

factors present in CM.

Camelina meal used in the present study was found to

contain glucosinolates, fibre, non-starch polysaccharides

and phytate. Glucosinolates in particular have been found

to affect nutrient digestibility, growth performance and

Figure 2 Principal component analysis of amino acid profiles from

the muscle tissue of both rainbow trout and Atlantic salmon fed

the 0% CM diet and either the 14% CM or the 16% CM diets.

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


digestion physiology in salmonids and have been reported

when feeding canola protein concentrate (Thiessen et al.

2004; Burr et al. 2013) and rapeseed meal (Burel et al.

2000). Glucosinolates specifically induced hypothyroidism

in rainbow trout (Thiessen et al. 2004). Phytate signifi-

cantly reduces protein utilization and growth, and

increased levels of dietary phytate induced structural

changes in pyloric caeca of rainbow trout (Forster et al.

1999). Non-starch polysaccharides in plant ingredients can

interfere with nutrient availability and utilization, gut mor-

phology and physiology and can negatively affect growth

performance in salmonids (Sinha et al. 2011). These com-

pounds may also affect the overall palatability of the diet,

which can result in reduced feed intake as was observed in

salmon. These antinutritional factors may have caused the

reduced gross energy digestibility as observed in both rain-

bow trout (57%) and Atlantic salmon (62%) (Fraser &

Anderson 2012). Therefore, the presence of antinutritional

factors may have resulted in decreased feed intake and

reduced digestibility, in both salmon and rainbow trout

fed, increasing amounts of CM in the diet, which ultimately

led to decreased growth performance in both species.

Although the known presence of antinutritional factors

and nutrient digestibility coefficients provides evidence on

the effect of CM on growth performance, a direct histologi-

cal analysis on the digestive physiology of salmonids fed

CM is necessary to confirm the negative impact that high

levels of CM can have nutrient absorption and growth

performance.

Fatty acid tissue composition was influenced by

CM inclusion in salmon more than in trout. In salmon,

significant increases in 18:3n-3 and 22:1n-9 and a decrease

in 22:6n-3 were observed with dietary CM inclusion,

whereas in trout, few significant differences in the fatty acid

profile among groups were observed. However, when the

fatty acid profiles were visualized in the MDS plot, these

subtle yet significant changes in the fatty acid profiles

caused distinct clustering of individuals according to treat-

ment and species. Species differences in fatty acid composi-

tion are expected, considering the difference in age class

and environment between rainbow trout and salmon in this

study; yet inclusion of CM in the diet similarly affected the

fatty acid profile of both species. These results were also

confirmed by SIMPER and ANOSIM; the fatty acid profile in

the muscle tissue of individuals fed the highest level of CM

was most different to those fed the FM based diet. How-

ever, the relationship is complicated as a result of changes

in fish oil (FO) and FM amounts in the diet, which did not

necessarily decrease as CM was included in the diet. Die-

tary lipid generally increased with CM inclusion, which

resulted in higher total lipid in the muscle. Therefore, lipid

and fatty acid composition in the tissue is likely a result of

both CM inclusion and FO/FM levels in the diet, not sim-

ply an effect of CM alone. Although the 21% CM diet

contained the highest level of CM, it also supplied the

greatest amount of FO and FM, even compared to the

control. This revealed the complicated relationship between

CM and FO/FM inclusion in the diet. Including 21% CM

in the diet had a detrimental effect on growth in rainbow

trout despite having the highest level of FO/FM in the

diet and there being few significant differences in tissue

composition.

(a) (b)

Figure 3 (a) Principal components analysis (PCA) of fatty acids, amino acids and lipid classes in the muscle tissue of rainbow trout fed

diets containing camelina meal (7% CM, 14% CM, 21% CM). (b) PCA of fatty acids, amino acids and lipid classes in the muscle tissue of

Atlantic salmon fed diets containing camelina meal (7% CM, 14% CM, 21% CM).

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


The amino acid tissue composition in rainbow trout did

not differ among groups fed different levels of dietary CM;

the stability of the amino acid profile despite dietary

changes may explain why rainbow trout growth perfor-

mance was not affected by lower levels of CM. In salmon,

increased dietary CM resulted in increased tissue amounts

of threonine, aspartic acid, glutamic acid and phenylala-

nine, and decreased amounts of lysine and tyrosine, despite

increasing amounts in the diet. The PCA illustrated species

differences in amino acid profiles, as would be expected,

but did not show a division between individuals fed a diet

with CM versus the control. Therefore, the amino acid pro-

file of both rainbow trout and salmon was not distinguish-

able in those that were fed a diet with or without CM

inclusion. Amino acid vectors for tyrosine, phenylalanine,

aspartic acid and glutamic acid were directed towards

Atlantic salmon individuals; these amino acids increased or

decreased significantly with CM inclusion. Although ala-

nine, leucine and valine were not significantly different

among groups of salmon, they increased slightly in the

muscle tissue with CM inclusion and their vectors were

associated with the Atlantic salmon amino acid profiles.

Because concentrations of some of the amino acids were

not significantly different in ANOVA or ANOSIM analyses, cor-

relations with amino acids and individuals in the plot may

indicate synergistic relationships with other factors in both

species.

In order to further understand the relationships among

nutrients in relation to growth performance, a PCA was

conducted which included lipids, fatty acids and amino

acids in the muscle tissue of fish fed CM diets. Both PCA

plots showed associations among lipids, fatty acids and

amino acids that serve common purposes related to growth

and survival. In the rainbow trout plot, several associations

are obvious, such as 18:2n-6 and 18:1n-9, and are likely

associated with the prevalent fatty acids in CM. Phospho-

lipids, sterols, 22:6n-3 and 20:4n-6 have important implica-

tions for cell membrane structure and fluidity in fish

(Sargent et al. 2002). Total lipids, neutral lipids and leucine

are also located together. Leucine stimulates muscle protein

synthesis and is rapidly mobilized from white and red mus-

cle during exercise (Nakashima et al. 2007), while total lip-

ids (and specifically neutral lipids) are critical for energy

production. Specific growth rate and weight gain were

located near tyrosine, histidine, 20:1n-9 and glutamic acid,

all of which serve as an energy substrate (Li et al. 2009).

In the Atlantic salmon PCA, total lipid, fatty acid and

amino acid composition had few similarities with the rain-

bow trout PCA, again accentuating the difference between

these species. Weight gain and specific growth rate are cen-

tred in the plot and closely located to glycine, tyrosine, leu-

cine and 16:1n-7. Again, tyrosine, leucine and

monounsaturated fatty acids serve feed intake, growth and

energy-related functions. A group containing 22:6n-3 and

lysine is also relevant because both key compounds

decreased in the tissue after CM was included in the diet.

A decrease in lysine and an increase in 18:3n-3 may be two

key factors in determining the growth performance of sal-

mon in this experiment. It is surprising that lysine

decreased in the muscle tissue because free lysine-HCl was

included in all diets to balance lysine requirements for sal-

monids (2% of the diet) (NRC 2011). It is possible that the

lysine added to the diets was not well absorbed by salmon,

as crystalline amino acids may be absorbed at a different

rate than protein-bound amino acids, which can reduce the

efficiency of uptake (Peres & Oliva-Teles 2005) and ulti-

mately affect the growth performance (Dabrowski et al.

2010). However, these changes did not necessarily impact

the food-related chemistry of the fillets. The sensory panel

results revealed no difference in the appearance, odour and

texture of salmon fillets that were fed 0% CM or 24% CM

diets. Although it is unlikely that CM would be included at

24% of a commercial diet due to poor growth perfor-

mance, CM inclusion should not impact consumer percep-

tion of the final product.

The results of this experiment reveal how different spe-

cies within the same family may metabolize similar diets

containing a novel ingredient. Rainbow trout seemed to

tolerate dietary CM (up to 14% CM) better than Atlantic

salmon, which were impacted by the lowest level of dietary

CM (8%) in this experiment. There is evidence that Atlan-

tic salmon may be more sensitive to plant meal ingredients

compared to rainbow trout, in terms of digestibility and

nutrient utilization, which has been suggested in previous

studies (Refstie et al. 2000; Krogdahl et al. 2004; Burr

et al. 2013). New research focused on the intestinal histol-

ogy of salmon fed CM diets may directly explain why die-

tary CM was not tolerated. Processing CM into a protein

concentrate or with heat treatment may remove antinutri-

tional compounds; therefore, a feeding experiment with

concentrated or treated CM is needed. The use of multivar-

iate statistics provided a useful visual representation of the

dynamics between fatty acids, amino acids and growth per-

formance after salmonids were fed CM diets that altered

the biochemical composition of their tissues. These rela-

tionships proved to be complicated and synergistic; this

highlights the challenges in supplying non-marine ingredi-

ents in aquaculture feeds and also the difficulties in formu-

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


lating diets to include terrestrial ingredients that are not

nutritionally equivalent to FM and FO.

This study was supported by Genome Atlantic, Atlantic

Canada Opportunities Agency (ACOA)-Atlantic Innova-

tion Fund (AIF) and Research and Development Corpora-

tion of Newfoundland (RDC). The authors would like to

acknowledge Dr. Matthew Rise for conceptual contribution

to the project; Danny Boyce and the Joe Brown Aquacul-

ture Research Building staff for maintenance of the Atlan-

tic salmon; Dr. Marije Booman, Dr. Tiago Hori, Dr. Kim

Johnstone, Charles Yu Feng and Xi Xue for fish sampling,

and Northeast Nutrition for supplying the majority of the

feed ingredients used in the study.

Burel, C., Boujard, T., Escaggre, A., Kaushik, S., Bœuf, G., Mol,

K., Van der Geyten, S. & Kuehn, E. (2000) Dietary low-gluco-

sinolate rapeseed meal affects thyroid status and nutrient utiliza-

tion in rainbow trout (Oncorhynchus mykiss). Br. J. Nutr., 83,

653–664.Burr, G., Wolters, W., Barrows, F. & Donkin, A. (2013) Evalua-

tion of a canola protein concentrate as a replacement for fish-

meal and poultry by-product meal in a commercial production

diet for Atlantic salmon (Salmo salar). Int. Aquat. Res., 5, 5.

Carpenter, R., Lyon, D. & Hasdell, T. (2000) The relationship of

physiology and psychology to sensory analysis. In: Guidelines

for Sensory Analysis in Food Product Development and Quality

Control, (Carpenter, R., Lyon, D. & Hasdell, T. eds), 2nd edn,

pp. 16–19. Aspen Publishers Inc, Gaithersburg, MD, USA.

Collins, S., Overland, M., Skrede, A. & Drew, M. (2013) Effect of

plant protein sources on growth rate in salmonids: meta-analysis

of dietary inclusion of soybean, pea and canola/rapeseed meals

and protein concentrates. Aquaculture, 400, 85–100.Crampton, V. & Carr, I. (2012) Spotlight 5: Fish Forever, pp.

5–14. EWOS publication, Bergen, Norway. http://www.ewos.

com/wps/wcm/connect/ewos-content-group/ewos-group/resources/

ewos-spotlight (Cited 18 May 2014).

Dabrowski, K., Zhang, Y.F., Kwasek, K., Hliwa, P. & Os-

taszewska, T. (2010) Effects of protein-, peptide- and free amino

acid-based diets in fish nutrition. Aquacult. Res., 41, 668–683.FAO (Food and Agriculture Organization). (1999). Examples of

attributes of fishery products used in sensory evaluation. In:

Report of the twenty-third session of the codex committee on fish

and fishery products (Annex I), Bergen, Norway: Food and Agri-

culture Organization of the United Nations.

Forster, I., Higgs, D., Dosanjh, B., Rowshandeli, M. & Parr, J.

(1999) Potential for dietary phytase to improve the nutritive

value of canola protein concentrate and decrease phosphorus

output in rainbow trout (Oncorhynchus mykiss) held in 11°Cfresh water. Aquaculture, 179, 109–125.

Fraser, J. & Anderson, D. (2012) Digestibility of Camelina sativa

seed and its by-products by Atlantic cod (Gadus morhua), rain-

bow trout (Oncorhynchus mykiss), and Atlantic salmon (Salmo

salar). Aquaculture Canada Abstracts 2012: New Frontiers:

Bridging Technology and Economic Growth. 27–30 May 2012.

Hixson, S., Parrish, C. & Anderson, D. (2014a) Changes in lipid

biochemistry of farmed rainbow trout (Oncorhyncus mykiss) fed

diets containing camelina oil (Camelina sativa) as a full replace-

ment of fish oil. Lipids, 49, 97–111.Hixson, S., Parrish, C. & Anderson, D. (2014b) Full substitution

of fish oil with camelina oil, with partial substitution of fish meal

with camelina meal, in diets for farmed Atlantic salmon (Salmo

salar) and its effect on tissue lipids and sensory quality. Food

Chem., 157, 51–61.Jobling, M. (1983) Growth studies with fish- overcoming the prob-

lems of size variation. J. Fish Biol., 22, 153–157.Krogdahl, A., Sundby, A. & Olli, J. (2004) Atlantic salmon (Salmo

salar) and rainbow trout (Oncorhynchus mykiss) digest and

metabolize nutrients differently. Effects of water salinity and die-

tary starch level. Aquaculture, 229, 335–360.Krogdahl, A., Penn, M., Thorsen, J., Refstie, S. & Bakke, A.

(2010) Important anti-nutrients in plant feedstuffs for aquacul-

ture: an update on recent findings regarding responses in salmo-

nids. Aquacult. Res., 41, 333–444.Li, P., Mai, K., Trushenski, J. & Wu, G. (2009) New developments

in fish amino acid nutrition: towards functional and environmen-

tally oriented aquafeeds. Amino Acids, 37, 43–53.Matthaus, B. (1997) Antinutritive compounds in different oilseeds.

Lipid/Fett., 5, 170–174.Morais, S., Edvardsen, R., Tocher, D. & Bell, B. (2012) Tran-

scriptomic analyses of intestinal gene expression of juvenile

Atlantic cod (Gadus morhua) fed diets with Camelina oil as

replacement for fish oil. Comp. Biochem. Physiol. B, 161, 283–293.

Morrison, W. & Smith, L. (1964) Preparation of fatty acid methyl

esters and dimethylacetals from lipids with boron fluoride-meth-

anol. J. Lipid Res., 5, 600–608.Mudalkar, S., Golla, R., Ghatty, S. & Reddy, A. (2014) De novo

transcriptome analysis of an imminent biofuel crop, Camelina sa-

tiva L. using Illumina GAIIX sequencing platform and identifi-

cation of SSR markers. Plant Mol. Biol., 84, 159–171.Nakashima, K., Yakabe, Y., Ishida, A., Yamazaki, M. & Abe, H.

(2007) Suppression of myofibrillar proteolysis in chick skeletal

muscles by a-ketoisocaproate. Amino Acids, 33, 499–503.National Research Council (NRC) (2011) Nutritional Require-

ments of Fish and Shrimp. National Academies Press, Washing-

ton, DC, USA.

Parrish, C. (1987) Separation of aquatic lipid classes by Chro-

marod thin-layer chromatography with measurement by Iatro-

scan flame ionization detection. Can. J. Fish Aquat. Sci., 44,

722–731.Parrish, C. (1999) Determination of total lipid, lipid classes and

fatty acids in aquatic samples. In: Lipids in Freshwater Ecosys-

tems (Arts, M. & Wainman, B. eds), pp. 4–10. Springer-Verlag,New York, NY, USA.

Peres, H. & Oliva-Teles, A. (2005) The effect of dietary protein

replacement by crystalline amino acid on growth and nitrogen

utilization of turbot Scophthalmus maximus juveniles. Aquacul-

ture, 250, 755–764.Putnam, D., Budin, J., Field, L. & Breene, W. (1993) Camelina: a

promising low output oilseed. In: New Crops (Janick, J. &

Simon, J. eds), pp. 314–332. John Wiley and Sons, New York,

NY, USA.

Refstie, S., Korsoen, O., Storebakken, T., Baeverfjord, G., Lein, I.

& Roem, A. (2000) Differing nutritional responses to dietary

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


http://www.ewos.com/wps/wcm/connect/ewos-content-group/ewos-group/resources/ewos-spotlight



soybean meal in rainbow trout (Oncorhynchus mykiss) and

Atlantic salmon (Salmo salar). Aquaculture, 190, 49–63.Sargent, J., Tocher, D. & Bell, G. (2002) The lipids. In: Fish

Nutrition (Halver, J. & Hardy, R. eds), pp. 181–247. Academic

Press Inc., San Francisco, CA, USA.

Shearer, K. (2000) Experimental design, statistical analysis and

modelling of dietary nutrient requirement studies for fish: a criti-

cal review. Aquacult. Nutr., 6, 91–102.Sinha, A., Kumar, V., Makkar, H., De Boeck, G. & Becker, K.

(2011) Non-starch, polysaccharides and their role in fish nutri-

tion – a review. Food Chem., 127, 1409–1426.Thiessen, D., Maenz, D., Newkirk, R., Classen, H. & Drew, M.

(2004) Replacement of fishmeal by canola protein concentrate in

diets fed to rainbow trout (Oncorhynchus mykiss). Aquacult.

Nutr., 10, 379–388.Ye, C. (2014) Evaluation of camelina (Camelina sativa) byproducts

fed to Atlantic salmon (Salmo salar) in practical diets. M.Sc.

Thesis. Appendix A. pp. 100. Dalhousie University, Truro, NS,

Canada.

Additional Supporting Information may be found in the

online version of this article:

Figure S1. Cluster analysis of fatty acid profiles from

Atlantic salmon and rainbow trout fed 0% CM, 14% CM

or 16% CM diets.

Table S1. Lipid class and fatty acid composition of control

and experimental diets for rainbow trout and Atlantic sal-

mon smolts.

Table S2. Amino acid composition of control and experi-

mental diets for rainbow trout and Atlantic salmon smolts.

Table S3. Growth performance of rainbow trout, analyzed

by quadratic regression and confirmed by ANOVA.

Table S4. Analysis of similarities (ANOSIM) and similarity

of percentages (SIMPER) results based on muscle tissue

fatty acid composition in rainbow trout and Atlantic sal-

mon fed diets containing fish meal or increasing levels of

camelina meal.

Table S5. Analysis of similarities (ANOSIM) and similarity

of percentages (SIMPER) results based on muscle tissue

amino acid composition1 in rainbow trout and Atlantic sal-

mon fed diets containing fish meal or increasing levels of

camelina meal.

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inclusion of camelina meal as a protein source in diets ...€¦ · rainbow trout (oncorhynchus...

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