See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/286031300

Rainbow trout as a model for nutrition and nutrient metabolism studies

Article · January 2013

CITATIONS READS
15 1,193

3 authors:

Stephane Panserat Sadasivam Kaushik

French National Institute for Agriculture, Food, and Environment (INRAE) Universidad de Las Palmas de Gran Canaria
233 PUBLICATIONS   6,671 CITATIONS    424 PUBLICATIONS   19,992 CITATIONS   

SEE PROFILE SEE PROFILE

Françoise Médale
French National Institute for Agriculture, Food, and Environment (INRAE)
216 PUBLICATIONS   7,087 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

ARRAINA View project

Endocrine dosrupting effect of genistein in fish View project

All content following this page was uploaded by Sadasivam Kaushik on 15 February 2018.

The user has requested enhancement of the downloaded file.

In: Trout: From Physiology to Conservation ISBN: 978-1-62417-009-6
Editors: Sergio Polakof and Thomas W. Moon © 2013 Nova Science Publishers, Inc.

Chapter 8

RAINBOW TROUT AS A MODEL FOR NUTRITION

AND NUTRIENT METABOLISM STUDIES

Stéphane Panserat, Sadasivam Kaushik

and Françoise Médale
INRA, Nutrition, Métabolisme, Aquaculture, Saint-Pée-sur-Nivelle, France

ABSTRACT
The rainbow trout [Oncorhynchus mykiss] is a carnivorous [piscivorous] species that
shows many particularities with respect to nutrition. The rainbow trout is a poor user of
dietary carbohydrates illustrated by a clear glucose intolerance phenotype after
carbohydrate intake. Moreover, rainbow trout have a high requirement for protein (the
major macronutrient in diet), with amino acids being largely used for energy production
by catabolism or through the production of fatty acids and glucose. Finally, trout energy
metabolism is related to their poikilothermic lifestyle as with most aquatic animals. This
chapter will focus primarily on protein, energy and glucose metabolism as related to the
nutritional regulation of intermediary metabolism in tissues including liver, muscle, fat
and intestine. In addition, one potential nutritional strategy to improve the efficiency of
nutrient utilization in rainbow trout will be detailed.

1. INTRODUCTION
Salmonids are amongst the most highly cultured fish groups worldwide. For fish
aquaculture to be successful and sustainable, fish nutrition is one of the main criteria to be
improved [1, 2]. It is the intent of this chapter to summarize what features are specific to the
nutrition of salmonids with a specific focus on the rainbow trout, Oncorhynchus mykiss. This
chapter will not provide an extensive description of trout nutrition, but it will describe
specific metabolic and nutritional criteria which characterize the salmonids with respect to the
feed utilization.


E-mail address: panserat@st-pee.inra.fr
132 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

2. NUTRITION IN RAINBOW TROUT: A PREAMBULE

Salmonids are predatory fish in nature and are placed at a highest trophic level of the
food chain of the aquatic environment. These fish species are classified as carnivorous
animals. In this context, they represent a very interesting model of animal nutrition because,
up to now, few strict carnivorous animals have been extensively studied with the exception of
cats [3], mink [4], raptors such as owls [5] and, indeed, the piscivorous fish species. Data on
nutrient requirements of fish have been recently updated by NRC [2]. The nutrition of
salmonids [Atlantic salmon, Salmo salar; rainbow trout] has received significant attention as
they are the predominant species reared and reviews on the nutrition of rainbow trout are
available [6, 7]. Aquafeeds have been developed over the past thirty years for salmonids in
aquaculture; these formulations are based on the well-known nutrient requirements identified
for these species. It is imperative to develop new aquafeeds in order to replace dietary fish
meal and fish oil [originating from threatened marine resources] by either plant ingredients or
other ingredients in the context of the development of sustainable aquaculture [8, 9]. This
evolution of dietary composition necessitates an extensive knowledge of the nutritional
biochemistry [intermediary metabolism] of these aquatic animals [2]. Indeed, studies
regarding the macro- and micro-nutrient requirements of rainbow trout at different
developmental stages are numerous and are well described in a large number of reviews [2,
10-12]; these will not be discussed in the present chapter, but the reader should refer to these
other papers. Instead, we will develop and describe some targeted characteristics linked to the
nutrition of the rainbow trout, which can explain why this animal is so interesting in
nutritional science. In particular, we will describe energy use in fish, which is linked to their
poikilothermy and ammoniotelism. In addition, we will detail the role played by dietary
proteins and amino acids in carnivorous fish. And finally, we will examine the low dietary
carbohydrate use by rainbow trout. In the end, we will provide an example of the new feeding
strategies that are being investigated to improve nutrient use in trout, called nutritional
programming.

3. ENERGY METABOLISM AND THE CONTRIBUTION

OF MACRONUTRIENTS AS ENERGY SOURCES IN RAINBOW TROUT

Rainbow trout, as all organisms, need energy for maintaining vital functions and
renewing tissues, for physical activity, for growth and the development of specific tissues
such as the gonads. These energy needs are met under aerobic conditions by the oxidative
catabolism of organic compounds (carbohydrates, fats and proteins). When animals are
fasted, they draw energy from body reserves provided during times of feeding. When fed,
energy is provided by the macromolecules derived from the digestion of ingested foods.
When energy supplied by food exceeds maintenance needs it is stored as body reserves, and
ultimately leads to growth. Once absorbed, amino acids arising from protein digestion, fatty
acids and glycerol from the digestion of dietary lipids and glucose resulting from digestion of
polysaccharides are either incorporated into cell structures after biosynthesis, or catabolized
in exergonic reactions thereby producing energy that is used to meet energy needs. The
potential energy of organic molecules is released by oxidative pathways that transform the C-
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 133

C bonds of these molecules to CO2 and H2O. Pathways of energy production are common to
all animals:

 formation of acetyl-CoA from fatty acid oxidation or the breakdown of amino acids
or pyruvate from glycolysis;
 incorporation of acetyl-CoA into the tricarboxylic acid cycle with the production of
CO2, reduced nucleotides and hydrogen atoms; and,
 the oxidation of reduced nucleotides by the electron transport chain to molecular
oxygen, a process that is accompanied by capturing the energy of electron transport
in the phosphorylation of ADP, generating ATP, the cellular fuel.

Despite these common pathways, energy metabolism of rainbow trout differs from that of
mammals in several aspects and in particular in terms of both the quantitative requirements
and the specific substrates used for energy production. In addition, while the routes are
common, their regulation by nutrients is often specific.

3.1. Low Energy Needs in Relation to Poikilothermy

Energy needs in the fasting trout are about 30-80 kJ/kg body weight/day depending
mainly upon water temperature. Because fish are poikilothermic animals [i.e., body
temperature vary close to that of the surrounding water], their metabolic rate is strongly
affected by water temperature [13]. When water temperature decreases, fish body temperature
decreases accordingly so that basal energy expenditure is reduced. Maintenance energy
requirements of trout are estimated as 75 to 100 kJ/kg/day [14]; i.e., about 10- to 20-fold
lower than those of endothermic terrestrial vertebrates [15]. The fact that poikilothermic
animals do not maintain internal body temperatures above ambient temperatures represents a
major savings of energy. Thus, rainbow trout are a relevant model for comparative studies
regarding the role of proteins involved in non-shivering thermogenesis such as uncoupling
proteins (UCP) [16]. Excretion of nitrogen wastes directly in the form of ammonia (more than
75% of the total nitrogen wastes) mainly through passive processes contributes also to the
low metabolic rate of fish compared to terrestrial animals and birds that expend significant
energy for the synthesis and excretion of urea and uric acid.

3.2. Differences in Macronutrients Used as Energy Sources

In addition to differences in quantitative energy requirements, energy metabolism of

carnivorous fish such as trout differs from that of mammals and birds by the proportion of
macronutrients that are utilized as substrates for energy production. While glucose is a
principal source of cellular energy in mammals, it is a relatively minor energy source in trout.
In contrast, proteins/amino acids and lipids are the preferred substrates for energy production.
The relative proportions of nutrients that are catabolized vary greatly depending on the
balance of macronutrients in the diet.
134 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

3.3. Energy Production from Amino Acids Catabolism

Trout [as many fish species] differ from terrestrial vertebrates in that they oxidize a large
proportion of the amino acids supplied by the digestion of dietary protein [or breakdown of
body protein when trout are fasted] for energy production and that the main end product of
nitrogen catabolism is ammonia. Nitrogen is removed through the process of transdeami-
nation that involves amino transferases [aspartate and alanine amino transferase being
quantitatively the most important] and glutamate dehydrogenase that plays a major role in the
oxidation of amino acids. The amino acid is transformed into an α-ketoacid with the
concomitant synthesis of glutamate from α-ketoglutarate. The α-ketoglutarate is then
regenerated by the deamination of glutamate by glutamate dehydrogenase. Glutamate, being
transaminated by many amino acids, represents a crossroad of the use of amino acids for
energy production. The liver is the main site of this process in trout and other teleosts.
Glutamate dehydrogenase is present in higher activities in the liver of fish than in mammals
[17]. Ammonia (NH3) from the deamination of amino acids is converted into the less toxic
ammonium ion (NH4+) which is excreted across the gills [75%] and in the urine (25%) [18].
N-ammonia excretion is measured as a relevant indicator of protein catabolism. The
regulation of this pathway is described in section 4 below. The carbon skeleton remaining
after nitrogen removal can be used for gluconeogenesis via oxaloacetate or be oxidized within
the tricarboxylic acid cycle. Due to the major role of α-ketoglutarate, the activity of the
tricarboxylic acid cycle is regulated not only by the activity of citrate synthase [that controls
the entry of acetyl-CoA into the cycle] but also by entry at the level of α-ketoglutarate.
Consequently, isocitrate dehydrogenase (IDH) that produces α-ketoglutarate through the
oxidative decarboxylation of isocitrate while converting NADP+ to NADPH and NAD+ to
NADH is a key-enzyme in the process. In trout liver as in other tissues, activity of NADP+-
IDH exceeds that of NAD+-IDH; activities of NAD+-IDH are present but low possibly as a
result of the instability of this enzyme [19]. Because amino-acid catabolism diverts amino
acids from protein synthesis and generates nitrogen waste, major efforts have been made to
limit this pathway. In rainbow trout this has been achieved by increasing the dietary supply of
non-protein energy in the form of lipids [20].

3.4. Dietary Lipid as an Efficient Energy Source

Most of the basic pathways of lipid metabolism including digestion and absorption, lipid
transport, lipogenesis and β-oxidation are essentially the same in trout as they are in mammals
[21, 22]. The main products of dietary lipid digestion are free fatty acids along with partial
acylglycerols, lyso-phospholipids, cholesterol, and fatty alcohols. Once solubilized or
emulsified in bile salt micelles, these products diffuse mainly by passive process to the
intestinal mucosa where uptake into the enterocytes occurs. In these intestinal mucosal cells,
the predominant fate of the absorbed free fatty acids is re-esterification with glycerol and
lyso-phospholipids to form triacylglycerols and phospholipids. Free cholesterol is transported
from the mucosal cells and fatty alcohols are oxidized into the corresponding fatty acids in
the epithelial cells.
Although some free fatty acids can be transported bound to albumin-like proteins, the
majority of lipids are transported in the blood in the form of lipoproteins that are similar to
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 135

those found in mammals. These include the chylomicrons that transport the majority of
absorbed dietary lipid away from the intestine, the very low density lipoproteins (VLDL) that
have a high lipid:protein ratio, the low density lipoprotein (LDL), and the high density
lipoproteins (HDL) that have a low lipid:protein ratio.
Clearance of lipoproteins from the plasma is mediated by their respective receptors.
Activity and expression of LDL-R are modulated by the dietary fatty acids [23].
Uptake of fatty acids by tissues is mediated by lipoprotein lipase (LPL) that hydrolyzes
triacylglycerols of triacylglycerol-rich lipoproteins (chylomicrons and VLDL) and generates
fatty acids that are taken up by tissues. Intracellular transport of fatty acids is facilitated by
tissue-specific cytoplasmic fatty acid binding proteins (FABP) that bind long-chain fatty acids
[21].
Fatty acid catabolism is a major source of energy in trout through -oxidation that occurs
in peroxisomes and ultimately mitochondria of all cells whereas lipogenesis occurs mainly in
the liver. After activation, fatty acids are transported into the mitochondria in the form of fatty
acylcarnitine esters formed through the action of the carnitine acyltransferase complex,
converted back into fatty acyl-CoA derivatives that then enter a round of dehydrogenation,
hydration, and cleavage steps to produce acetyl-CoA and NADH. Several steps within this
process are tightly regulated by nutrient and hormones. However, the key step seems to be the
entry of fatty-acyls into mitochondria that is controlled by carnitine-acyltransferase 1 and the
second dehydrogenation step that is mediated by the hydroxy-acyl CoA dehydrogenase
[24-26].
Acetyl-CoA generated through fatty-acid -oxidation can then be metabolized via the
tricarboxylic acid cycle to produce more NADH which then provides metabolic energy in the
form of ATP through the processes of electron transport and oxidative phosphorylation. In
vivo studies investigating fatty acid deposition show that, generally, the higher the
concentration of a fatty acid in the diet, the lower its retention, suggesting that increased
concentrations lead to increased oxidation except for C22:6 n-3 [DHA] that is preferentially
retained in tissues [27, 28].
Salmonids including rainbow trout are among the animals that can consume the highest
quantity of fat (as much as humans). Excess dietary lipid is deposited in adipose cells of the
intraperitoneal [perivisceral] adipose tissue, and to some extent as a layer between the skin
and flesh [subcutaneous adipose tissue] and in the muscle (intramuscular fat).
The majority of body lipid depots arise from dietary lipids, the contribution of lipogenesis
being limited [24, 29]. However new lipids can be synthetized from acetyl-CoA itself formed
in the mitochondria from the oxidative decarboxylation of pyruvate [carbohydrate source] or
the oxidative degradation of some amino acids (protein source). The lipogenic pathway is
catalyzed by the cytosolic fatty acid synthetase (FAS) multi-enzyme complex which produces
the saturated fatty acids 16:0 and 18:0. Monounsaturated fatty acids are then produced by
microsomal stearoyl CoA desaturase (SCD) or 9 desaturase, generating 18:1n-9.
Fatty acid elongases produce longer chain saturated and monounsaturated fatty acids,
such as 20:0 and 20:1n-9. However, the polyunsaturated C18 fatty acids cannot be
synthesized de novo by any vertebrate and must be obtained in the diet.
Rainbow trout are able to synthesize the long chain highly unsaturated fatty acids such as
EPA and DHA from linolenic acid (C18:3 n-3) contrary to some marine species, despite the
genes coding for 5-6 desaturases and elongase have been reported [30].
136 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

3.5. The Use of Glucose as an Energy Supplier Depends on the Tissue

Glycolysis, which converts glucose to pyruvate with the formation of ATP, is the route of
glucose catabolism in trout tissues. Glycolytic reactions are reversible and can therefore
operate in the direction of gluconeogenesis (3 carbon conversion to 6 carbon glucose) with
the exception of three steps catalyzed by hexokinase (formation of glucose-6-P),
phosphofructokinase (formation of fructose-1,6-bisphosphate) and pyruvate kinase
(conversion of phosphoenolpyruvate to pyruvate). These three enzymes are considered to be
the limiting and unidirectional steps in glycolysis. Under aerobic conditions, pyruvate, after
decarboxylation to acetyl-CoA, supplies carbon to the tricarboxylic acid cycle. Under
anaerobic conditions, which prevail in the white muscle of trout during intense activity,
pyruvate is converted to lactate by the cytosolic enzyme lactate dehydrogenase (LDH). This
route of pyruvate utilization is of paramount importance for energy production in the white
muscle of trout that supports high-speed swimming White muscle that represents
approximately 95% of the total skeletal muscle mass of trout is composed of anaerobic
glycolytic fibers. The reversible conversion of pyruvate to lactate allows quick bursts of
anaerobic energy supported by muscle glycogen reserves [31]. LDH generates lactate
ensuring the re-oxidation of NADH generated within glycolysis thus allowing such activities
as pursuing and catching prey, chasing competitors, escaping predators [in the wild] to
continue [32]. As in most vertebrates, LDH is present as several isozymes that are tissue
specific. However, the isozymes of trout have particular characteristics. The temperature
required for maximum activity is much lower than that required for the mammalian enzyme
and the enzyme-substrate binding is different [33, 34]. In contrast, aerobic energy production
through oxidative pathways prevails in the red muscle. This tissue [less than 5% of the total
skeletal muscle mass of trout] is composed of aerobic, oxidative fibers that are recruited for
supporting sustained moderate swimming. According to Richards et al. [35], ATP used for
contraction in the red muscle of rainbow trout under sustained swimming is generated by the
oxidation of approximately 45% glucose, 35% lipid and 20% protein. The high energy yield
of oxidative pathways (compared to anaerobic energy production), allows a more effective
use of muscle reserves. In addition some recent studies with trout muscle cells in primary
culture support the idea that mechanisms regulating the entry of glucose into skeletal muscle
may be stimulated by AMPK activation [36]. Activities of the mitochondrial enzymes citrate
synthase (CS) and cytochrome oxidase have been shown to reflect the aerobic pathway in fish
muscles [31, 37] and are measured to assess the contribution of the aerobic pathways to
energy production in the tissues. Although all enzymes of glycolysis have been detected in
trout tissues, in the liver, the major role of glycolysis is to provide precursors for the
biosynthesis of various molecules over the production of pyruvate for oxidation. It may be
that the species that have evolved on high-lipid food sources are more likely to have a poor
utilization of dietary carbohydrate.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 137

4. NUTRITIONAL REGULATION OF PROTEIN

AND AMINO ACID METABOLISM

4.1. Dietary Protein Requirements of Rainbow Trout Are High

As mentioned, critical assessments of nutrient requirements of rainbow trout have been

assessed over the past two decades [6, 7] and especially with reference to protein and amino
acids [11, 38, 39]. In line with what is known for most aquatic animals, the dietary protein
requirement of rainbow trout is high, the optimum protein level in feeds depending upon
dietary digestible energy content. Dietary digestible protein (DP) requirements of rainbow
trout are 36-38% of the diet when there is an adequate supply of non-protein digestible energy
(DE). Under farming conditions, improving dietary protein utilization for body protein
growth with low impacts on the environment in terms of nitrogen loads is of utmost
importance. Optimizing amino acid supply with respect to requirements is especially
significant in the case of animals living in an aquatic environment.
Using the protein efficiency ratio which reflects unit wet weight gain per unit protein
intake as one criterion, it was found relatively early that the maximum efficiency in most
teleost fishes is observed at relatively low dietary protein levels, similar to what is observed in
terrestrial omnivores [38, 40]. When protein gain is considered as the criterion, under optimal
farming conditions, protein retention relative to dietary protein intake is higher (40-50%) in
rainbow trout than in many other teleosts or in most of terrestrial farmed animals. Such an
improvement in protein utilization was made possible over the decades through optimizing
the overall protein supply with regard to the overall demands of the organism meeting the
quantitative needs for all the ten indispensable amino acids (IAA).

4.2. Protein and IAA Requirements for Maintenance and Growth

Rainbow trout, as most fish known today, have a dietary requirement for the same ten
IAA as almost all terrestrial animals including humans. A comparison of data between IAA
requirements of juvenile rainbow trout and that of a pure terrestrial carnivore such as the cat
(see Table 1) shows that while the total IAA requirements of rainbow trout are lower than that
of the kitten, when expressed per unit DE the requirements of rainbow trout are much higher
than that of the kitten. This has indeed been recognized for some time and much effort has
been put into optimizing the DP/DE ratio in trout (Cho and Kaushik, 1990).
From a practical point of view, the ideal dietary protein appears to be the one that has the
IAA profile closely resembling that of the whole body of the fish. Since the whole body
amino acid profile does not seem to vary as a function of age in rainbow trout [41], little
differences in the IAA requirements over the growth cycle would be expected. The
observation that the ideal amino acid profile for trout (in fact, fish in general), is that of the
whole body of trout has practical implications in formulating feeds with ideal amino acid
profiles.
138 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

Table 1.Data on indispensible amino acid needs (expressed as g/100g DP or as mg/kJ

DE) of rainbow trout compared to those of the kitten

Juvenile rainbow trout Kitten

g/100g DP mg/kJ DE g/100g DP mg/kJ DE
Arg 4.2 0.9 10.0 0.48
His 2.2 0.5 3.0 0.14
Ile 3.1 0.6 5.0 0.24
Leu 4.2 0.9 12.0 0.57
Lys 6.7 1.4 8.0 0.38
Met + Cys 3.1 0.6 7.5 0.36
Phe + Tyr 5.0 1.0 8.5 0.41
Thr 3.1 0.6 7.0 0.34
Trp 0.8 0.2 1.5 0.07
Val 3.3 0.7 6.0 0.29
Sum IAA 40.0 8.2 68.5 3.18
Data for rainbow trout recalculated from NRC (2011) and those for kitten from NRC (1986). DP:
digestible protein; DE: digestible energy.

This also possibly explains why fishmeal has remained the most appropriate and the
major dietary protein source in the feeds for farmed rainbow trout. Knowledge on the IAA
needs also provides objective data for supplementing diets with adequate amounts of the
potentially limiting amino acids in developing diets in which a large portion of the dietary
fish meal is substituted with other protein sources be they plant or animal. Much progress has
been made in developing feeds for trout with very low levels of fish meal [39, 42, 43]. The
importance of the ratio between indispensable to dispensable amino acids (IAA/DAA ratio)
has also been stressed and a ratio of 55:45 appears optimal for rainbow trout [44].
When we consider the relative proportions of protein used for maintenance and growth,
data available to date indicate that the maintenance component of the protein requirement for
growth of fish is low (15 to 30% of total protein need), compared to that of terrestrial
vertebrates (>50%), leaving more scope for growth for a given protein intake in the case of
fish. With regard to the IAA requirements for maintenance, the most complete set of data
available today for any fish is that for the rainbow trout [45].
Data on sulphur amino acids of rainbow trout were found to be about two-fold higher
than those recorded for terrestrial omnivores [46]. This possibly also explains the higher need
for exogenous amino acid supply for growth and protein accretion in fish in comparison to
terrestrial monogastrics. The maintenance requirements for arginine were found to be low in
several teleosts including rainbow trout [47].
Since obligatory nitrogen or amino acid losses under protein-free feeding conditions can
be assumed to reflect the minimum physiological needs for IAA, based on data from whole
body amino acid losses in rainbow trout fed a protein-free diet over 4 weeks Fournier et al.
[47] were able to calculate the endogenous losses in amino acids in rainbow trout to be about
350 mg/kg/day providing data on body protein degradation rates.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 139

4.3. Improving Nitrogen Utilization Is a Major Objective

One explanation for the high dietary protein requirements of rainbow trout, as in almost
all teleosts, is that a significant amount of energy is derived from nitrogenous compounds
[48] and amino acids that serve as preferential oxidative substrates [49]. Although rainbow
trout have an apparent high dietary protein requirement for expressing their maximum growth
potential, the efficiency with which dietary protein is converted into edible tissue protein is
generally higher in rainbow trout than in most food producing animals. Since under farming
conditions, the major objective is to increase the supply of proteins for human consumption,
over the past two decades specific strategies have been adopted to enhance protein retention:
by optimizing the quantitative supply of dietary proteins, by improving the amino acid
balance of the diets, and by increasing the supply of DE from non-protein sources.
As regards the nutritional control of protein and amino acid metabolism, some general
concepts put forward very early prevail even today. Rainbow trout, when confronted with
changes in dietary protein levels, appear not to show changes in their amino acid catabolism.
In trout, the activities of enzymes of amino acid catabolism are only slightly affected by
changes in dietary protein levels; data available today suggest that there is more an effect of
dietary protein levels or of overall protein intake on intermediary metabolism [50-52] rather
than on direct amino acid catabolism per se.
This situation is somewhat similar to what is seen in terrestrial carnivores such as the cat
which also exhibits a high dietary protein requirement, high obligatory nitrogen losses and a
limited capacity to control amino acid catabolism [53]. This lack of control of amino acid
catabolism as affected by dietary protein levels is indeed considered to be one major reason
for the high protein requirements of fish [54].
Data based on studies on N balance as well as protein synthesis rates as affected by water
temperature show that any effect on protein accretion is mainly mediated through an increase
in voluntary feed intake and the proportion of the different segments of N balance remain the
same [55]. An increase in the supply of some of the dietary IAA is known to increase protein
synthesis and/or deposition in terrestrial animals. No such proteogenic effect has so far been
demonstrated in rainbow trout receiving an excess dietary leucine [56].

4.4. Efforts towards Reducing Nitrogen Excretion

A significant proportion of ingested nitrogen is lost as soluble nitrogenous wastes arising

from branchial and urinary excretion. Much work on the production and excretion of
nitrogenous catabolites has been undertaken with rainbow trout, showing that the most
prominent excretory products are ammonia [about 80 to 85% of total nitrogen excreted] and a
small amount of urea [57]. There is in general a direct relationship between nitrogen intake
and ammonia excretion but not such a clear effect on urea excretion [58]. Dietary amino acids
absorbed in excess of the amounts required for maintenance and growth are eliminated as
waste products affecting the immediate environment. Very early it was shown with rainbow
trout that ammonia excretion can be significantly reduced by increasing the non-protein
energy supply through digestible carbohydrates or fat with a consequent reduction in the
DP/DE ratio [59]. There is also a good correlation between dietary protein quality (expressed
in terms of the EAA (Essential Amino Acids) index) and N excretion. Attempts to improve
140 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

nitrogen utilization through non-protein nitrogen sources such as urea have not been
successful. Dietary urea for instance is readily absorbed in the digestive tract of juvenile trout
but is totally excreted with no beneficial effect on nitrogen balance [60]. Unlike other
vertebrates, urea excretion in rainbow trout is considered to be a reflection of the maintenance
turnover of arginine [61] and nucleic acids rather than the end product of the ornithine-urea
cycle. Besides the liver which is generally recognized as the active site of ureagenesis, it has
also been shown that urea cycle enzyme activities in skeletal muscle tissue can account for a
significant portion of total urea excretion in juvenile and adult trout [62].
Increased N utilization will decrease nitrogenous wastes into the environment. Based on
studies with rainbow trout, models relating N excretion to N intake have also been proposed,
very much applicable to farming conditions in order to reduce environmental impacts [63].
While ammoniotelism confers some advantages to fish from the energetic point of view,
excess ambient ammonia has adverse effects in all aquaculture systems. Hence, there is a
necessity to optimize the amino acid supply for the reduction of ammonia excretion into the
environment, since the excess supply of amino acids is more likely to increase ammonia
excretion, without having any beneficial effect on protein growth.
Under the current global context of stagnation of world fishery catches and increasing
demands for fishmeal for fish farming, replacement of fishmeal with other protein sources in
aquatic animal feeds is an important issue. When dealing with fish meal replacement, it is
worth looking into possible alternative protein sources which are sufficiently rich in protein
as is fishmeal. Such fishmeal substitutes are limiting in one or more IAA [11]. There is clear
evidence now that dietary amino acid supplementation can reduce the overall protein level in
the feeds of rainbow trout, provided proper attention is paid to the IAA profile. Our
knowledge of the interactions between different amino acids, their roles in other metabolic
pathways beyond protein accretion and as signaling molecules are little known and research
along these lines are warranted from a comparative perspective between different fish and
other terrestrial monogastrics.

5. POOR METABOLIC USE OF DIETARY CARBOHYDRATES

IN RAINBOW TROUT

5.1. No Dietary Carbohydrates Requirement in Rainbow Trout

Glucose metabolism and glucose homeostasis are highly dependent of the feeding status
of the animal. In fed as well as unfed animals, glucose will be used directly as an energy
source in many tissues through glycolysis. But in fed animals dietary glucose in excess will
be either stored as glycogen mainly in muscle and liver or converted into fatty acids in liver
and fat tissue in order to be used latter [during the fasted state] for energy purposes. Feeding
rainbow trout with carbohydrates [highly present in plant ingredients] has been studied
primarily for aquaculture purposes [10, 64, 65]. However it is generally accepted that rainbow
trout do not have a specific requirement for dietary glucose [2] because they are able to
survive and grow with diets devoid of carbohydrates. It is interesting to study dietary
carbohydrates in rainbow trout because it is theoretically a good strategy to incorporate
dietary glucose as ingredients in order to decrease levels of fishmeal [rich in proteins] and
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 141

fish oil [rich in lipids] in new aquafeeds. Carbohydrates are found as a major component of
many plant products, generally in the form of starch. Unfortunately, even though the level of
incorporation of digestible dietary carbohydrates has been largely tested in rainbow trout, it is
always limited at a level lower than 20% (20 g of carbohydrates per 100 g of diet) [64, 66].
Indeed, carbohydrate inclusion in pellets is limited due to a significant decrease in growth
parameters associated with a postprandial persistent hyperglycemia (up to 20 mM) in rainbow
trout fed with more than 20% carbohydrates [10, 66, 67]. Sometimes a detrimental effect on
liver (―fatty liver‖) is also observed [10, 66, 68]. These data suggest a problem in metabolic
use of dietary glucose in rainbow trout.

5.2. Digestion of Dietary Carbohydrates in Rainbow Trout: Positive Effects

of Technological Treatments of Diets

The first step in the nutrition process is the intestinal digestion of nutrients. Carbohydrate
sources can be arranged schematically into two categories: the digestible saccharides
(monosaccharide, disaccharides, polysaccharides (such as starch)) and the nondigestible
polysaccharides [non-starch polysaccharides (NSP)or dietary fiber) Rainbow trout (as many
fish species in aquaculture) cannot use NSP efficiently due to the absence of an adequate gut
microbiota for their digestion [69].
Concerning the others types of carbohydrates, monosaccharides [such as glucose] or
dissacharides (such as sucrose) can be incorporated directly in rainbow trout diets. Even
though they are efficiently absorbed by the gut, this is not a good practical solution for
aquafeed formulation in aquaculture due to their costs [64].
This is why the common source of dietary carbohydrates for fish nutrition is starch which
is common in plant product [10]. Actually, starch is highly digested by rainbow trout after
technological treatments of the pellets (such as extrusion and gelatinization). This process
produces a starch highly accessible to digestive enzymes expressed in trout including amylase
and maltase producing a nutrient that is more than 90% digestible [10, 64, 69].

5.3. Poor Use of Dietary Glucose by Rainbow Trout: Is It True for All Fish
Species?

Dietary carbohydrates are effectively digested and absorbed by the gut, but the challenge
for scientists is to understand and to improve the poor metabolism of dietary glucose by
rainbow trout [10, 64].
There is clearly a metabolic inability to use the level of dietary carbohydrates usually
found in mammalian diets [50% of carbohydrate in the diet for the rat, for example].
Importantly the low glucose use in rainbow trout fed carbohydrates is not observed for all fish
species; indeed, species at a low trophic level (such as the omnivorous common carp,
Cyprinus carpio and the Nile tilapia, Oreochromis niloticus) are able to use higher levels of
carbohydrates (50% in the diets)in contrast to the fish at the higher trophic level [the
piscivores or carnivores] fish such as the salmonids and many marine species [64, 66, 68].
142 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

We can hypothesize from an evolutionary perspective that fish having no carbohydrates

in their natural diet (i.e., piscivorous fish like rainbow trout) have lost their metabolic
capacities to deal with an increase in plasma glucose after a large carbohydrate intake. The
following paragraphs describe the potential bottleneck(s) at least in some tissues such as
intestine, pancreas, liver, muscle and fat tissue that may explain this poor dietary glucose use.
Indeed, determining which metabolic pathway is defective in rainbow trout is important to
understand the specificities of nutrition in this species.

5.4. Some of the Major Hypotheses to Explain Poor Dietary Carbohydrates

Use in Rainbow Trout

 Hypothesis I: poor hormonal response to dietary carbohydrate intake

After carbohydrate intake and in order to initiate glucose use and/or storage in target
tissues such as liver, muscle and fat tissue, plasma insulin levels increase in mammals. In fed
rainbow trout regardless of the macronutrient composition, there is an induction of insulin
secretion [70-72]. However, glucose is a poor insulin secretagogue compared to some amino
acids (e.g. arginine) in many fish species (e.g., see 73, 74) The relatively low level of insulin
secretion after carbohydrate intake could be related to the concomitant high levels of
somatostatin secretion [from the pancreas] as described by some authors [75, 76],
somatostastin being an inhibitor of insulin secretion. Atypical molecular regulation of the
ATP-dependent inward rectifying potassium channels in the pancreas of rainbow trout fed
with carbohydrates could also be another explanation for the poor insulin secretion after
glucose intake [77]. Further studies are however necessary in order to better understand the
nutritional regulation of the secretion for some key (pancreatic) hormones (such as glucagon)
involved in regulation of nutrient (glucose) metabolism. However, the hypoglycemia effect of
insulin injection in the regulation of glycemia in rainbow trout appears to be similar to the one
observed in mammals and is linked to the induction of the insulin signalling pathways (IRS
phosphorylation, Akt phosphorylation) in liver and muscle [78-82].

 Hypothesis II: poor regulation of nutrient signalling in nutrient-sensitive

tissues after carbohydrate intake

Regulation of intermediary metabolism through some metabolic sensors is needed to

control intracellular glucose use. The study of two of these metabolic sensors, AMP-activated
protein kinase (AMPK) and the mammalian Target of Rapamycin [mTOR], has been recently
analysed in rainbow trout by our laboratory [79, 83, 84]. Indeed, after feeding, there is a
decrease in the phosphorylation of AMPK [due to the decrease of the AMP/ATP ratio] in
liver and a concomitant increase in mTOR phosphorylation [due to the concomitant effects of
insulin and amino acids] in liver and muscle as reported in mammals [79, 83, 84]. The
pharmacological activator of hepatic AMPK - AICAR, an analogue of AMP - in vivo and in
vitro (hepatocyte cultures) has the expected induction of glucose catabolism in trout liver
suggesting a similar role for this enzyme in the regulation of intermediary metabolism [84]. In
the same manner, the inhibition of the mTOR pathway using the specific inhibitor rapamycin
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 143

also effects the molecular regulation of intermediary metabolism in trout hepatocytes, in

particular on gluconeogenic and glycolytic enzyme gene expression [85]. Thus mTOR and
AMPK are sensors that globally regulate post-carbohydrate feeding in both mammals and
trout. Moreover, the possible over-activation of mTOR in muscle and liver due to the high
levels of postprandial plasma amino acids in carnivorous fish could have a negative effect on
insulin action and glucose metabolism as has been demonstrated in mammals [86]. Recently
we observed a specific induction of mTOR activation and an inhibition in AMPK
phosphorylation in rainbow trout liver when fed with carbohydrates compared to no
carbohydrates [Kamalam et al.; unpublished data]. These data suggest a system of nutrient
sensing sensitive to carbohydrate intake in rainbow trout very similar to that observed in
mammals.

 Hypothesis III: poor regulation of intermediary metabolism by dietary

carbohydrates in liver, muscle and fat tissue

The consequences of hormones and nutrients are ultimately the regulation of the
intermediary metabolism. A number of studies have detailed the effects of carbohydrate
intake on glucose metabolism especially in rainbow trout [10, 66].
Dietary glucose storage in liver. After feeding carbohydrates, there is an up-regulation of
the hepatic glycolytic pathway linked to increased activities of glucokinase(GK),
phosphofructokinase and pyruvate kinase in rainbow trout [87-89]. The induction of the
activity of hepatic GK after carbohydrate feeding is always associated with a dramatic
induction of glucokinase gene expression [similar to an on-off regulation] showing
unambiguously the possibility of trout adapting to carbohydrate intake. Surprisingly, and in
contrast to mammals, this induction is strictly linked to carbohydrate intake and not to insulin
levels in vivo [80, 81] or in vitro [80, 85]. The storage of glucose in excess as glycogen also
occurs in trout liver as demonstrated by the higher levels of glycogen and higher activities of
glycogen synthase in trout fed with carbohydrates compared to the fish fed without
carbohydrates [66]. The pentose phosphate pathway also responds at both molecular and
biochemical levels in trout liver [66, 90], suggesting that this metabolic pathway may
function for the production of NADPH required for anabolic pathways (including
lipogenesis). Even though lipogenesis is not directly linked to glucose metabolism,
lipogenesis plays an important role in glucose homeostasis for the conversion of glucose [in
excess] into fatty acids. However, the role of hepatic lipogenesis on glucose homeostasis is
marginal due to its low induction by carbohydrate intake [90] even though it is sensitive to
insulin treatment [26]. This poor induction of lipogenesis in rainbow trout fed carbohydrates
could be one explanation for the poor glucose homeostasis in these fish species: an induction
of lipogenesis in liver through a drug [metformin] [90, 91] or after genetic selection [92] is
associated with a lower postprandial glycemia in fed rainbow trout. Moreover, a recent study
has demonstrated that lipogenesis in white adipose tissue of rainbow trout is induced by
insulin suggesting a potential role for this tissue in the use of dietary glucose [26]. This area
needs further study.
Regulation by dietary carbohydrates of hepatic glucose production. In rainbow trout fed
with carbohydrates [87, 89], inhibition of key hepatic gluconeogenic enzymes is generally not
observed at either the molecular or activity levels in contrast to what is observed in mammals
144 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

[93] and in some glucose tolerant fish species [94]. The absence of a clear reduction in
hepatic gluconeogenesis is another possible explanation for poor dietary carbohydrate use by
rainbow trout [10].
Dietary glucose use in white muscle and adipose tissue. In white muscle, no clear
regulation of glucose metabolism (glycolysis, glycogenesis) by dietary carbohydrates is
detected in rainbow trout and other fish species [10, 66, 95]. Indeed, even though there is an
insulin-sensitive glucose transporter 4 (Glut4) in trout muscle [96, 97], glycolytic rates remain
constant and low as reflected by the absence of higher hexokinase and pyruvate kinase (PK)
enzyme activities in muscle of trout fed with compared to without carbohydrates [10, 64, 66,
90]. These data suggest a low adaptive capacity of fish muscle to accept dietary glucose even
though a small increase in glycogen content occurs in fish fed with carbohydrates and PK
activities seem always to be high. The low capacity to use dietary glucose in muscle is
supported by the fact that glucose phosphorylation [the first limiting step of glycolysis] is
always low as shown by the constant low activities of muscle hexokinase in rainbow trout
[10, 64, 66]. Moreover the low number of insulin receptors in muscle of the trout compared
with glucose tolerant fish such as the common carp [98] is also an indicator of a poor capacity
of muscle to use dietary glucose. On the other hand, after feeding carbohydrates, plasma
glucose can also be used by the perivisceral adipose tissue. A recent study in trout reported
glucose transport and glucose phosphorylation and lipogenesis are present in this tissue [26]
but to date, no study has examined the regulation of glucose metabolism by dietary
carbohydrates in this tissue.

6. TOWARDS IMPROVING EFFICIENCY OF NUTRIENT UTILIZATION

IN RAINBOW TROUT: A NEW FEEDING STRATEGY,
THE NUTRITIONAL PROGRAMMING

Many studies have tried to suppress the (metabolic) bottlenecks that may decrease the
capacities of rainbow trout to adapt to new diets in aquaculture (see above). One such strategy
is to modify diet composition through new combinations of feed ingredients. In this context,
some targeted strategies have been developed. One example was to decrease as much as
possible the levels of proteins and fat which, at high levels, have negative impacts on dietary
glucose use by acting on hepatic gluconeogenesis (increasing it) and lipogenesis [decreasing
it] [99, 100]. Here, we will describe an approach that is promising for aquaculture and is
called nutritional programming.
Definition of the nutritional programming. Studies in some mammals and insects (bees)
show that dietary influences exerted at critical early life stages (neonatal nutrition, post-natal
(weaning]) nutrition) may have long-term consequences on physiological functions in later
life [101-103]. This phenomenon, known as nutritional programming, is largely studied in
mammals for the understanding of diseases such as the metabolic syndrome or diabetes [103,
104]. Development represents a period of rapid change in gene expression during which
environmental cues [such as nutrition] may induce persistent changes in the phenotype of an
organism. Variation in nutrition during early life induces different phenotypes that are
contingent on the timing, duration and nature of the nutritional challenge.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 145

Examples of nutritional programming in mammals. Animal models have been valuable in

proving proof of concept for nutritional programming at the fetal stage. A number of animal
models, including both large (e.g. sheep) and small (e.g. rats and mice) animals have been
established to investigate the effects of the early environment on long-term health [102, 104,
105]. In the rat, maternal protein restriction, maternal calorie restriction, maternal anemia,
intrauterine artery ligation, and fetal exposure to glucocorticoids result in features of the
metabolic syndrome in the offspring. The phenotypic outcomes of these insults are very
similar, suggesting that they act through common pathways. Moreover, studies have shown
that alterations in the quality of nutrition (such as levels of carbohydrate intake) in early life
(at postnatal stages), without changes in the total caloric intake, can also function as an
independent environmental cue for induction of metabolic programming effects leading to
chronic hyperinsulinemia and adult-onset obesity [101, 106, 107]. Therefore, altered
nutritional experiences (under, over or modified nutrition) in the immediate postnatal period
are important and independent cues for inducing metabolic programming effects.
Possible mechanisms to explain the nutritional programming. Possible biological
mechanisms for storing the nutritional programming events until adulthood include adaptive
changes in gene expression (epigenetic phenomenon linked to the methylation of DNA and
histones, miRNAs), preferential clonal selection of adapted cells in programmed tissues, and
programmed differential proliferation of tissue cell types [102, 103, 108]. Presently one of the
main (known) mechanisms behind the nutritional programming is the epigenetic changes
which corroborates with distinct gene expression profiles in specific tissues [109]; epigenetics
changes, in particular DNA methylation, underlie the induction and persistence of altered
phenotypes. Differentiation in the expression of the genome of the honey bee by nutrition is
probably the clearest example of induction of alternative phenotypes and epigenotypes by
nutrition in early life [110].
What is known in nutritional programming in rainbow trout. Epigenetic changes
associated with chromatin remodelling and regulation of gene expression underlie the concept
of metabolic programming [103], but these processes have not been deciphered in fish larvae.
Nevertheless, certain metabolic functions [intestinal transport, glucose and fatty acid
metabolism…] in juvenile fish also depend on specific nutritional signals during critical larval
periods, demonstrating that the concept of metabolic programming also exists in fish [111,
112].
The concept of early-programming raises the interesting possibility of directing specific
metabolic pathways or functions in juvenile fish, for example to improve the use of
substitutes to fish meal and oil, and hence to promote sustainability in aquaculture. Two first
and original approaches in nutritional programming have recently been tested in teleosts in
order to improve two crucial metabolic pathways: glucose metabolism [111] and the synthesis
of n-3 highly unsaturated fatty acids (n-3 HUFA) [112, 113].
Geurden et al. [111] reported that an acute but short (few days) nutritional stimulus
consisting of a hyperglucidic diet, applied in early post-hatch life of rainbow trout induced a
persistent positive effect on carbohydrate digestion revealed at the juvenile stages by a higher
persistent expression of amylase and maltase in early-stimulated fish compared to controls
(Figure 1).
These preliminary data and those obtained in other fish species (see [112, 113] for
programming in saltwater fish) are promising and may offer new nutritional strategies in
order to improve use of new ingredients in diets of rainbow trout.
146 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

« Commercial diet »
Very High Carbohydrates diet = 65% dextrin
During 2 months
First feeding: 3 days

In intestine In intestine
Genes Type of regulation Genes Type of regulation
α-amylase Up-regulation 4-fold α-amylase (GI) Up-regulation 1.7-fold
Maltase Up-regulation 2.2-fold Maltase (GI) Up-regulation 1.5-fold

Figure 1. Persistent higher gene expression of carbohydrate digestion enzymes compared to controls in
rainbow trout juvenile after a preliminary stimulus with high intake of carbohydrates [65% of dextrin]
at first feeding [adapted from 111].

CONCLUSION
This chapter has presented some of the main particularities of nutrient use in rainbow
trout, a very efficient user of dietary amino acids, a very poor user of dietary glucose
(carbohydrates), as well as some specificities linked to poïkilothermy(associated with low
energy use) and the aquatic environment (a reduced urea cycle, ammonium elimination across
the gills from the amino acid catabolism). It is clear that the rainbow trout is a very interesting
animal model to study nutrition. In particular, specific regulation of metabolism by amino
acids and glucose in this species could permit the development of new paradigms in
nutritional sciences. Moreover, and because the rainbow trout is also one of the main
aquaculture species, it is expected that this knowledge can improve the nutrition of fish in
aquaculture in the context of the development of new more sustainable diets for farmed fish.
Indeed, as an example, a new nutritional strategy based on nutritional programming could
improve the trout nutrition in the context of sustainable aquaculture.

REFERENCES
[1] FAO. Report of the twenty-ninth session of the comittee on fisheries2011.
[2] NRC. Nutrient Requirements of Fish and Shrimp. Washington: National Academy
Press; 2011.
[3] Verbrugghe A, Hesta M, Daminet S, Janssens GP. Nutritional modulation of insulin
resistance in the true carnivorous cat: a review. Crit. Rev. Food Sci. Nutr.
2012;52[2]:172-82.
[4] Bell J. Ferret nutrition. Vet. Clin. North Am. Exot. Anim. Pract. 1999;2(1):169-92.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 147

[5] Myers MR, Klasing KC. Low glucokinase activity and high rates of gluconeogenesis
contribute to hyperglycemia in barn owls (Tyto alba) after a glucose challenge. J. Nutr.
1999;129(10):1896-904.
[6] Hardy RW, Webster CD, Lin CE. Rainbow trout, Oncorhynchus mykiss. Nutrient
requirements and feeding in finfish for aquaculture. New York: CABI Publishing;
2002. p. 184-202.
[7] Cowey CB. Nutrition: estimating requirements of rainbow trout. Aquaculture.
1992;100(1-3):177-89.
[8] Naylor RL, Hardy RW, Bureau DP, Chiu A, Elliott M, Farrell AP, et al. Feeding
aquaculture in an era of finite resources. Proc. Natl. Acad. Sci. U.S.A.
2009;106(36):15103-10.
[9] Gatlin III DM, Barrows FT, Brown P, Dabrowski K, Gaylord TG, Hardy RW, et al.
Expanding the utilization of sustainable plant products in aquafeeds: a review.
Aquacult. Res. 2007;38(6):551-79.
[10] Panserat S. Nutritional regulation of gene expression for proteins involved in
metabolism in cultured fish: focus on dietary carbohydrates. In: Young G, Thorgaard
GH, Gatlin II DM, Craig S, editors. Current status of molecular research in aquaculture:
Wiley-Blackwell; 2009.
[11] Kaushik SJ, Seiliez I. Protein and amino acid nutrition and metabolism in fish: current
knowledge and future needs. Aquacult. Res. 2010;41(3):322-32.
[12] Corraze G, Kaushik S. Alimentation lipidique et remplacement des huiles de poisson
par des huiles végétales en pisciculture. Cah. Agric. 2009;18:112-8.
[13] Azevedo PA, Cho CY, Leeson S, Bureau DP. Effects of feeding level and water
temperature on growth, nutrient and energy utilization and waste outputs of rainbow
trout (Oncorhynchus mykiss). Aquat. Liv. Res. 1998;11[4]:227-38.
[14] Médale F, Guillaume J. Nutritional energetics. In: Guillaume J, Kaushik S, Bergot P,
Métailler R, editors. Nutrition and Feeding of Fish and Crustaceans. Chichester UK:
Springer Praxis Publishing; 2001. p. 59-79.
[15] Kaushik S, Médale F. Energy requirements, utilization and dietary supply to salmonids.
Aquaculture. 1994;124(1-4):81-97.
[16] Jastroch M, Wuertz S, Kloas W, Klingenspor M. Uncoupling protein 1 in fish uncovers
an ancient evolutionary history of mammalian nonshivering thermogenesis. Physiol.
Genomics. 2005;22(2):150-6.
[17] Walton MJ, Cowey CB. Aspects of intermediary metabolism in salmonid fish. Comp.
Biochem. Physiol. B-Biochem. Mol. Biol. 1982;73(1):59-79.
[18] Médale F, Brauge C, Vallee F, Kaushik SJ. Effects of dietary protein/energy ratio,
ration size, dietary energy source and water temperature on nitrogen excretion in
rainbow trout. Wat. Sci. Tech. 1995;31(10):185-94.
[19] Storey KB, Fields JHA. NAD+-linked isocitrate dehydrogenase in fish tissues. Fish
Physiol. Biochem. 1988;5(1):1-8.
[20] Rasmussen RS, Ostenfeld TH, McLean E. Growth and feed utilisation of rainbow trout
subjected to changes in feed lipid concentrations. Aquac. Int. 2000;8(6):531-42.
[21] Tocher DR. Metabolism and functions of lipids and fatty acids in teleost fish. Rev. Fish.
Sci. 2003;11(2):107-84.
[22] Sargent JR, Tocher DR, Bell JG. The lipids. In: Halver JE, Hardy RW, editors. Fish
nutrition. San Diego: Academic Press; 2002. p. 181-257.
148 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

[23] Richard N, Kaushik S, Larroquet L, Panserat S, Corraze G. Replacing dietary fish oil by
vegetable oils has little effect on lipogenesis, lipid transport and tissue lipid uptake in
rainbow trout (Oncorhynchus mykiss). Br. J. Nutr. 2006;96(2):299-309.
[24] Kolditz C, Borthaire M, Richard N, Corraze G, Panserat S, Vachot C, et al. Liver and
muscle metabolic changes induced by dietary energy content and genetic selection in
rainbow trout (Oncorhynchus mykiss). Am. J. Physiol. Regul Integr Comp Physiol.
2008;294[4]:R1154-64.
[25] Morash AJ, Bureau DP, McClelland GB. Effects of dietary fatty acid composition on
the regulation of carnitine palmitoyltransferase [CPT] I in rainbow trout [Oncorhynchus
mykiss]. Comp. Biochem. Physiol. B-Biochem. Mol. Biol. 2009;152[1]:85-93.
[26] Polakof S, Medale F, Larroquet L, Vachot C, Corraze G, Panserat S. Insulin stimulates
lipogenesis and attenuates -oxidation in white adipose tissue of fed rainbow trout.
Lipids. 2011;46[2]:189-99.
[27] Bell JG, Tocher DR, Henderson RJ, Dick JR, Crampton VO. Altered fatty acid
compositions in Atlantic salmon (Salmo salar) fed diets containing linseed and
rapeseed oils can be partially restored by a subsequent fish oil finishing diet. J. Nutr.
2003;133[9]:2793-801.
[28] Thanuthong T, Francis DS, Senadheera SPSD, Jones PL, Turchini GM. LC-PUFA
biosynthesis in rainbow trout is substrate limited: use of the whole body fatty acid
balance method and different 18: 3n-3/18: 2n-6 ratios. Lipids. 2011:1-17.
[29] Kolditz CI, Plagnes-Juan E, Quillet E, Lefèvre F, Médale F. Changes in white muscle
transcriptome induced by dietary energy levels in two lines of rainbow trout
(Oncorhynchus mykiss) selected for muscle fat content. Br. J. Nutr. 2010;103[05]:
629-42.
[30] Zheng X, Seiliez I, Hastings N, Tocher DR, Panserat S, Dickson CA, et al.
Characterization and comparison of fatty acyl Delta6 desaturase cDNAs from
freshwater and marine teleost fish species. Comp. Biochem. Physiol. B-Biochem. Mol.
Biol. 2004;139[2]:269-79.
[31] Garenc C, Couture P, Laflamme MA, Guderley H. Metabolic correlates of burst
swimming capacity of juvenile and adult threespine stickleback (Gasterosteus
aculeatus). J. Comp. Physiol. B, Biochem. Syst. Environ. Physiol. 1999;169[2]:113-22.
[32] Sherwood Gv, Pazzia I, Moeser A, Hontela A, Rasmussen Jv. Shifting gears: enzymatic
evidence for the energetic advantage of switching diet in wild-living fish. Can. J. Fish.
Aquat. Sci. 2002;59[2]:229-41.
[33] Holmes RS, Markert CL. Immunochemical homologies among subunits of trout lactate
dehydrogenase isozymes. Proc. Natl. Acad. Sci. U.S.A. 1969;64(1):205.
[34] Tylicki A, Masztaleruk D, Strumilo S. Differences in some properties of lactate
dehydrogenase from muscles of the carp Cyprinus carpio and trout Salmo gairdneri.
Journal of Evolutionary Biochemistry and Physiology. 2006;42(2):143-7.
[35] Richards JG, Mercado AJ, Clayton CA, Heigenhauser GJ, Wood CM. Substrate
utilization during graded aerobic exercise in rainbow trout. J. Exp Biol. 2002;205(Pt
14):2067-77.
[36] Palstra AP, Planas JV. Fish under exercise. Fish Physiol Biochem. 2011;37(2):259-72.
[37] Guderley H, Gawlicka A. Qualitative modification of muscle metabolic organization
with thermal acclimation of rainbow trout, Oncorhynchus mykiss. Fish Physiol.
Biochem. 1992;10(2):123-32.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 149

[38] Cowey CB. Amino acid requirements of fish: a critical appraisal of present values.
Aquaculture. 1994;124(1-4):1-11.
[39] Kaushik S. Protein and amino acid nutrition of fish in comparison to terrestrial
vertebrates. In: Nunes AF, Portugal AV, Costa JP, Ribeiro JR, editors. Protein
Metabolism and Nutrition. Portugal: INIA; 1995. p. 47-56.
[40] Bowen SH. Dietary protein requirements of fishes-a reassessment. Can. J. Fish. Aquat.
Sci. 1987;44(11):1995-2001.
[41] Wilson RP, Cowey CB. Amino acid composition of whole body tissue of rainbow trout
and Atlantic salmon. Aquaculture. 1985;48(3-4):373-6.
[42] Kaushik SJ. Soybean products in salmonid diets. In: Lim C, Webster CD, Lee CS,
editors. Alternative protein sources in aquaculture diets. New York, USA: Haworth
Press; 2008. p. 261-79.
[43] Kaushik SJ, Hemre GI. Plant proteins as alternative sources for fish feed and farmed
fish quality. In: Lie O, editor. Improving farmed fish quality and safety. Cambridge,
UK: Woodhead Publishing Limited; 2008. p. 300-27.
[44] Green JA, Hardy RW, Brannon EL. The optimum dietary essential: nonessential amino
acid ratio for rainbow trout [Oncorhynchus mykiss], which maximizes nitrogen
retention and minimizes nitrogen excretion. Fish Physiol. Biochem. 2002;27(1):109-15.
[45] Rodehutscord M, Becker A, Pack M, Pfeffer E. Response of rainbow trout
(Oncorhynchus mykiss) to supplements of individual essential amino acids in a
semipurified diet, including an estimate of the maintenance requirement for essential
amino acids. J. Nutr. 1997;127(6):1166.
[46] Mambrini M, Kaushik S, editors. Effect of temperature on sulfur amino acid
requirements for maintenance and growth of juvenile rainbow trout. Proceedings of
thev7th International Symposium on Protein Metabolism and Nutrition; 1995; Santarem,
Portugal.
[47] Fournier V, Gouillou-Coustans MF, Metailler R, Vachot C, Guedes MJ, Tulli F, et al.
Protein and arginine requirements for maintenance and nitrogen gain in four teleosts.
Br. J. Nutr. 2002;87(5):459-69.
[48] Cowey CB, Luquet P, editors. Physiological basis of protein requirements of fishes.
Critical analysis of allowances. IV International Symposium on Protein Metabolism and
Nutrition. Les Colloques INRA; 1983.
[49] Van den Thillart G. Energy metabolism of swimming trout (Salmo gairdneri). J. Comp.
Physiol. B, Biochem. Syst. Environ. Physiol. 1986;156[4]:511-20.
[50] Kim KI, Grimshaw TW, Kayes TB, Amundson CH. Effect of fasting or feeding diets
containing different levels of protein or amino acids on the activities of the liver amino
acid-degrading enzymes and amino acid oxidation in rainbow trout (Oncorhynchus
mykiss). Aquaculture. 1992;107:89-105.
[51] Sánchez-Muros MJ, García-Rejón L, García-Salguero L, De la Higuera M, Lupiáñez
JA. Long-term nutritional effects on the primary liver and kidney metabolism in
rainbow trout. Adaptative response to starvation and high-protein, carbohydrate-free
diet to glutamate dehydrogenase and alanine aminotransferase kinetics. Int. J. Biochem.
1998;30:55-63.
[52] Moyano FJ, Cardenete G, De la Higuera M. Nutritive and metabolic utilization of
proteins with high glutamic acid content by the rainbow trout (Oncorhynchus mykis)
Comp. Biochem. Physiol. A Physiol. 1991;100(3):759-62.
150 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

[53] Taylor TP, Morris JG, Kass PH, Rogers QR. Maximal growth occurs at a broad range
of essential amino acids to total nitrogen ratios in kittens. Amino Acids.
1998;15(3):221-34.
[54] Cowey CB, Walton MJ, Halver J. Intermediary metabolism. Fish Nutrition. New York:
Academic Press; 1989. p. 259-329.
[55] Fauconneau B. The measurement of whole body protein synthesis in larval and juvenile
carp (Cyprinus carpio). Comparative Biochemistry and Physiology. Part B,
Comparative Biochemistry. 1984;78(4):845-50.
[56] Choo PS, Smith TK, Cho CY, Ferguson HW. Dietary excesses of leucine influence
growth and body composition of rainbow trout. J. Nutr. 1991;121(12):1932-9.
[57] Kaushik S, Cowey CB, Cho CY. Ammoniogenesis and dietary factors affecting
nitrogen excretion. Nutritional strategies and aquaculture waste. Guelph, Canada:
University of Guelph; 1991. p. 3-19.
[58] Kaushik SJ. Influence of nutritional status on the daily patterns of nitrogen excretion in
the carp (Cyprinus carpio L.) and the rainbow trout (Salmo gairdneri R) Reproduction,
Nutrition, Development. 1980;20[6]:1751-65.
[59] Kaushik SJ, de Oliva Teles A. Effect of digestible energy on nitrogen and energy
balance in rainbow trout. Aquaculture. 1985;50(1-2):89-101.
[60] Kaushik SJ, Dabrowski KR, Dabrowska H, Olah E, Luquet P. Utilization of dietary
urea in rainbow trout. Annals Nutr. Metab. 1983;27[2]:94-106.
[61] Fournier V, Gouillou-Coustans MF, Metailler R, Vachot C, Moriceau J, Le Delliou H,
et al. Excess dietary arginine affects urea excretion but does not improve N utilisation
in rainbow trout Oncorhynchus mykiss and turbot Psetta maxima. Aquaculture.
2003;217[1-4]:559-76.
[62] Todgham AE, Anderson PM, Wright PA. Effects of exercise on nitrogen excretion,
carbamoyl phosphate synthetase III activity and related urea cycle enzymes in muscle
and liver tissues of juvenile rainbow trout (Oncorhynchus mykiss). Comp. Biochem.
Physiol., Part A Mol. Integr. Physiol. 2001;129(2-3):527-39.
[63] Aubin J, Tocqueville A, Kaushik SJ. Characterisation of waste output from flow-
through trout farms in France: comparison of nutrient mass-balance modelling and
hydrological methods. Aquat. Liv. Res. 2011;24(01):63-70.
[64] Wilson RP. Utilization of dietary carbohydrate by fish. Aquaculture. 1994;124:67-80.
[65] Hemre GI, Bjornevik M, Beattie C, Björnsson BT, Hansen T. Growth and salt-water
tolerance of juvenile Atlantic salmon, Salmo salar, reared under different combinations
of dietary carbohydrate and photoperiod regime. Aquac. Nutr. 2002;8:23-32.
[66] Hemre GI, Mommsen TP, Krogdahl Å. Carbohydrates in fish nutrition: effects on
growth, glucose metabolism and hepatic enzymes. Aquac. Nutr. 2002;8:175-94.
[67] Moon TW. Glucose intolerance in teleost fish: fact or fiction? Comp. Biochem. Physiol.
B-Biochem. Mol. Biol. 2001;129[2-3]:243-9.
[68] Enes P, Panserat S, Kaushik S, Oliva-Teles A. Dietary carbohydrate utilization by
European sea bass (Dicentrarchus labrax L.) and gilthead sea bream (Sparus aurata L.)
juveniles. Rev. Fish. Sci. 2011;19(3):201-15.
[69] Krogdahl Å, Hemre GI, Mommsen TP. Carbohydrates in fish nutrition: digestion and
absorption in postlarval stages. Aquac. Nutr. 2005;11:103-22.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 151

[70] Baños N, Baró J, Castejón C, Navarro I, Gutiérrez J. Influence of high-carbohydrate

enriched diets on plasma insulin levels and insulin and IGF-I receptors in trout. Regul.
Pept. 1998;77:55-62.
[71] Capilla E, Médale F, Navarro I, Panserat S, Vachot C, Kaushik S, et al. Muscle insulin
binding and plasma levels in relation to liver glucokinase activity, glucose metabolism
and dietary carbohydrates in rainbow trout. Regul. Pept. 2003;110:123-32.
[72] Novoa MS, Capilla E, Rojas P, Baró J, Gutiérrez J, Navarro I. Glucagon and insulin
response to dietary carbohydrate in rainbow trout [Oncorhynchus mykiss]. Gen. Comp.
Endocrinol. 2004;139(1):48-54.
[73] Andoh T. Amino acids are more important insulinotropins than glucose in a teleost fish,
barfin flounder (Verasper moseri) Gen. Comp. Endocrinol. 2007;151[3]:308-17.
[74] Mommsen TP, Moon TW, Plisetskaya EM. Effecs of arginine on pancreatic hormones
and hepatic metabolism in rainbow trout. Physiol. Biochem. Zool. 2001;74(5):668-78.
[75] Ehrman MM, Melroe GT, Kittilson JD, Sheridan MA. Glucose-stimulated somatostatin
gene expression in the Brockmann bodies of rainbow trout (Oncorhynchus mykiss)
results from increased mRNA transcription and not from altered mRNA stability.
Zoolog. Sci. 2004;21(1):87-91.
[76] Eilertson CD, Sheridan MA. Pancreatic somatostatin-14 and somatostatin-25 release in
rainbow trout is stimulated by glucose and arginine. Am. J. Physiol. Regul. Integr.
Comp. Physiol. 1995;269(5 Pt 2):R1017-R23.
[77] Polakof S, Panserat S, Plagnes-Juan E, Soengas JL. Altered dietary carbohydrates
significantly affect gene expression of the major glucosensing components in
Brockmann bodies and hypothalamus of rainbow trout. Am. J. Physiol. Regul. Integr.
Comp. Physiol. 2008;295:R1077-R88.
[78] Castillo J, Codina M, Martínez ML, Navarro I, Gutiérrez J. Metabolic and mitogenic
effects of IGF-I and insulin on muscle cells of rainbow trout. Am. J. Physiol. Regul.
Integr. Comp. Physiol. 2004;286:R935-R41.
[79] Seiliez I, Gabillard JC, Skiba-Cassy S, García-Serrana D, Gutiérrez J, Kaushik S, et al.
An in vivo and in vitro assessment of TOR signaling cascade in rainbow trout (Oncor-
hynchus mykiss) Am. J. Physiol. Regul. Integr. Comp. Physiol. 2008;295(1):R329-35.
[80] Plagnes-Juan E, Lansard M, Seiliez I, Médale F, Corraze G, Kaushik S, et al. Insulin
regulates the expression of several metabolism-related genes in the liver and primary
hepatocytes of rainbow trout (Oncorhynchus mykiss). J. Exp. Biol. 2008;211(Pt
15):2510-8.
[81] Polakof S, Moon TW, Aguirre P, Skiba-Cassy S, Panserat S. Effects of insulin infusion
on glucose homeostasis and glucose metabolic gene expression in rainbow trout fed a
high-carbohydrate diet. J. Exp. Biol. 2010;213:4151-7.
[82] Caruso MA, Sheridan MA. New insights into the signaling system and function of
insulin in fish. Gen. Comp. Endocrinol. 2011;173(2):227-47.
[83] Lansard M, Panserat S, Seiliez I, Polakof S, Plagnes-Juan E, Geurden I, et al. Hepatic
protein kinase B (Akt)-target of rapamycin (TOR)-signalling pathways and
intermediary metabolism in rainbow trout (Oncorhynchus mykiss) are not significantly
affected by feeding plant-based diets. Br. J. Nutr. 2009;102(11):1564-73.
[84] Polakof S, Panserat S, Craig PM, Martyres DJ, Plagnes-Juan E, Savari S, et al. The
metabolic consequences of hepatic AMP-kinase phosphorylation in rainbow trout.
PLoS One. 2011;6[5]:e20228.
152 Stéphane Panserat, Sadasivam Kaushik and Françoise Médale

[85] Lansard M, Panserat S, Plagnes-Juan E, Seiliez I, Skiba-Cassy S. Integration of insulin

and amino acid signals that regulate hepatic metabolism-related gene expression in
rainbow trout: role of TOR. Amino Acids. 2010;39(3):801-10.
[86] Sengupta S, Peterson TR, Sabatini DM. Regulation of the mTOR complex 1 pathway
by nutrients, growth factors, and stress. Mol Cell. 2010;40(2):310-22.
[87] Panserat S, Médale F, Brèque J, Plagnes-Juan E, Kaushik S. Lack of significant long-
term effect of dietary carbohydrates on hepatic glucose-6-phosphatase expression in
rainbow trout (Oncorhynchus mykiss) J. Nutr. Biochem. 2000;11:22-9.
[88] Panserat S, Plagnes-Juan E, Brèque J, Kaushik S. Hepatic phosphoenolpyruvate
carboxykinase gene expression is not repressed by dietary carbohybrates in rainbow
trout (Oncorhynchus mykiss) J. Exp. Biol. 2001;204:359-65.
[89] Panserat S, Plagnes-Juan E, Kaushik S. Nutritional regulation and tissue specificity of
gene expression for proteins involved in hepatic glucose metabolism in rainbow trout
(Oncorhynchus mykiss) J. Exp. Biol. 2001;204 2351-60.
[90] Panserat S, Skiba-Cassy S, Seiliez I, Lansard M, Plagnes-Juan E, Vachot C, et al.
Metformin improves postprandial glucose homeostasis in rainbow trout fed dietary
carbohydrates: a link with the induction of hepatic lipogenic capacities? Am. J. Physiol.
Regul. Integr. Comp. Physiol. 2009;293(3):R707-R15.
[91] Polakof S, Moon TW, Aguirre P, Skiba-Cassy S, Panserat S. Glucose homeostasis in
rainbow trout fed a high-carbohydrate diet: metformin and insulin interact in a tissue-
dependent manner. Am. J. Physiol. Regul. Integr. Comp. Physiol. 2011;300(1):R166-
R74.
[92] Skiba-Cassy S, Lansard M, Panserat S, Medale F. Rainbow trout genetically selected
for greater muscle fat content display increased activation of liver TOR signaling and
lipogenic gene expression. Am. J. Physiol. Regul. Integr. Comp. Physiol.
2009;297(5):R1421-R9.
[93] Pilkis SJ, Granner DK. Molecular physiology of the regulation of hepatic gluco-
neogenesis and glycolysis. Annu. Rev. Physiol. 1992;54:885-909.
[94] Panserat S, Plagnes-Juan E, Kaushik S. Gluconeogenic enzyme gene expression is
decreased by dietary carbohydrates in common carp (Cyprinus carpio)and gilthead
seabream (Sparus aurata) Biochim. Biophys. Acta. 2002;1579(1):35-42.
[95] Felip O, Ibarz A, Fernandez-Borras J, Beltran M, Martin-Perez M, Planas JV, et al.
Tracing metabolic routes of dietary carbohydrate and protein in rainbow trout
(Oncorhynchus mykiss) using stable isotopes [[13C]starch and (15N]protein): effects of
gelatinisation of starches and sustained swimming. Br J. Nutr. 2011:1-11.
[96] Díaz M, Capilla E, Planas JV. Physiological regulation of glucose transporter(GLUT4)
protein content in brown trout [Salmo trutta] skeletal muscle. J. Exp. Biol. 2007;210(Pt
13):2346-51.
[97] Díaz M, Vraskou Y, Gutiérrez J, Planas JV. Expression of rainbow trout glucose
transporters GLUT1 and GLUT4 during in vitro muscle cell differentiation and
regulation by insulin and IGF-I. Am. J. Physiol. Regul. Integr. Comp. Physiol.
2009;296(3):R794-R800.
[98] Párrizas M, Baños N, Baró J, Planas J, Gutiérrez J. Up-regulation of insulin binding in
fish skeletal muscle by high insulin levels. Regul. Pept. 1994;53:211-22.
 Rainbow Trout as a Model for Nutrition and Nutrient Metabolism Studies 153

[99] Kirchner S, Kaushik S, Panserat S. Low protein intake is associated with reduced
hepatic gluconeogenic enzyme expression in rainbow trout (Oncorhynchus mykiss). J.
Nutr. 2003;133:2561-4.
[100] Figueiredo-Silva AC, Panserat S, Kaushik S, Geurden I, Polakof S. High levels of
dietary fat impair glucose homeostasis in rainbow trout. J. Exp. Biol. 2012;215[1]:
169-78.
[101] Patel MS, Srinivasan M, Laychock SG. Metabolic programming: Role of nutrition in
the immediate postnatal life. J. Inherit. Metab. Dis. 2009;32(2):218-28.
[102] Lucas A. Programming by early nutrition: an experimental approach. J. Nutr.
1998;128(2):401S-6S.
[103] Burdge GC, Lillycrop KA. Nutrition, epigenetics, and developmental plasticity:
implications for understanding human disease. Ann. Rev. Nutr. 2010;30:315-39.
[104] Gluckman PD, Hanson MA, Beedle AS. Early life events and their consequences for
later disease: a life history and evolutionary perspective. Am. J. Hum. Biol.
2007;19(1):1-19.
[105] Jones RH, Ozanne SE. Fetal programming of glucose-insulin metabolism. Mol. Cell
Endocrinol. 2009;297(1-2)4-9.
[106] Patel MS, Srinivasan M. Metabolic programming in the immediate postnatal life.
Annals Nutr. Metab. 2011;58(2)18-28.
[107] Patel MS, Srinivasan M. Metabolic programming due to alterations in nutrition in the
immediate postnatal period. J. Nutr. 2010;140(3):658-61.
[108] Warner M, Ozanne S. Mechanisms involved in the developmental programming of
adulthood disease. Biochem. J. 2010;427:333-47.
[109] Bonasio R, Tu S, Reinberg D. Molecular signals of epigenetic states. Science.
2010;330(6004):612.
[110] Kucharski R, Maleszka J, Foret S, Maleszka R. Nutritional control of reproductive
status in honeybees via DNA methylation. Science. 2008;319(5871):1827.
[111] Geurden I, Aramendi M, Zambonino-Infante J, Panserat S. Early feeding of carnivorous
rainbow trout (Oncorhynchus mykiss) with a hyperglucidic diet during a short period:
effect on dietary glucose utilization in juveniles. Am. J. Physiol. Regul. Integr. Comp.
Physiol. 2007;292(6):R2275-R83.
[112] Vagner M, Robin JH, Zambonino Infante JL, Person-Le Ruyet J. Combined effects of
dietary HUFA level and temperature on sea bass (Dicentrarchus labrax) larvae
development. Aquaculture. 2007;266(1-4)179-90.
[113] Vagner M, Zambonino Infante JL, Robin JH, Person-Le Ruyet J. Is it possible to
influence European sea bass (Dicentrarchus labrax)juvenile metabolism by a
nutritional conditioning during larval stage? Aquaculture. 2007;267(1-4):165-74.
[114] NRC. Nutrient requirements of cats. Washington, D.C.: National Academy Press; 1986.

View publication stats