You are on page 1of 9

Aquaculture 305 (2010) 150–158

Contents lists available at ScienceDirect

Aquaculture
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e

Energy, protein and amino acid requirements for maintenance and efficiency of
utilization for growth of Atlantic salmon post-smolts determined using increasing
ration levels
Ståle J. Helland ⁎, Bjarne Hatlen, Barbara Grisdale-Helland
Aquaculture Protein Centre, CoE and Nofima Marin, N-6600 Sunndalsøra, Norway

a r t i c l e

i n f o

Article history:
Received 23 October 2009
Received in revised form 25 February 2010
Accepted 15 April 2010
Keywords:
Atlantic salmon Salmo salar
Energy utilization efficiency
Protein utilization efficiency
Amino acid utilization efficiency
Ration level
Maintenance requirement

a b s t r a c t
Ration levels were used to determine the digestible protein (DP) and amino acids as well as digestible (DE)
and metabolizable energy (ME) requirements of Atlantic salmon post-smolts for maintenance and the
efficiency of utilization for gain above maintenance. The salmon (96 g) were held in seawater at a
temperature of 10 °C and were fasted for 28 days or fed incremental ration levels of a fishmeal-based diet for
51 days. The composition of gain was determined by the comparative slaughter technique.
The relationships between intake of digestible or metabolizable nutrients or energy and gain of those
nutrients or energy were linear (R2 ≥ 0.98) between zero intake and maximum voluntary intake. The DE and
ME requirements for maintenance were 31.5 and 30.6 kJ kg− 0.8 d− 1, respectively, and the efficiency of
utilization of DE and ME for gain were 0.80 ± 0.01 and 0.82 ± 0.01, respectively. The efficiency of utilization
for gain of DP was 0.64 ± 0.01 and that of the sum of digestible amino acids 0.57 ± 0.02. The efficiency
of utilization of the individual amino acids for gain varied from 0.45 for tryptophan to 0.65 for histidine.
The maintenance requirement for DP was 117.5 mg kg− 0.7 d− 1 and for digestible sum of amino
acids 204.0 mg kg− 0.7 d− 1. The maintenance requirements for the individual amino acids varied from
4.3 mg kg− 0.7 d− 1 for methionine and tryptophan to 20.9 mg kg− 0.7 d− 1 for leucine. The results suggest
more emphasis should be directed towards determining the maintenance requirements and the efficiency of
utilization for gain above maintenance for the individual amino acids and the sum of amino acids rather than
for crude protein.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction
The California Net Energy System (CNES) presented by Lofgreen
and Garrett (1968) was the first system developed using comparative
slaughter techniques which separated the metabolic requirements of
growing ruminants between maintenance and that needed for body
weight gain. The net energy (NE) value of the feed was determined
using these two requirements. Data for this system was generated by
feeding feedstuffs at two or more ration levels and determining
energy balance. The data were extrapolated to estimate the energy
loss at zero energy intake, and this was assumed to be equivalent to
the heat production of fasting animals. As described by Lofgreen and
Garrett (1968), NE required for maintenance (NEm) is equivalent to
the heat production of the fasting animal; whereas, the metabolizable
energy (ME) requirement for maintenance (MEm) is calculated as ME
intake (MEi) when energy gain is zero. Lofgreen and Garrett (1968)
noted in their earlier work that the efficiency of NE used for weight

⁎ Corresponding author. Tel.: +47 93418897; fax: +47 71695301.
E-mail address: stale.helland@nofima.no (S.J. Helland).
0044-8486/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2010.04.013

gain was linear and therefore constant. In the 1960s, Blaxter and
colleagues (reviewed by Ferrell and Oltjen, 2008) initiated the use of
two linear relationships describing the efficiency of ME use for
maintenance (slope of line relating energy loss and MEi, denoted km)
and for body growth (slope of line relating energy gain and MEi,
denoted kf) (Blaxter et al., 1966) (the latter will be denoted kMEg in
this paper). This method of using different ration levels to estimate
the ME requirements of animals has been used for carp, rainbow trout,
African catfish, Nile tilapia, and Atlantic salmon (Huisman, 1976;
Hogendoorn, 1983; Meyer-Burgdorff et al., 1989; Azevedo et al., 1998,
2005; Bureau et al., 2006). A modification of this method, where
digestible energy (DE) intake (DEi) replaces MEi, and estimates of the
maintenance requirement for digestible energy (DEm) and the
efficiency of utilization of DE for growth above maintenance (kDEg)
have been made, has been used with gilthead seabream, European sea
bass, white grouper, rainbow trout, silver perch, Atlantic cod and
barramundi (Lupatsch et al., 1998, 2001a,b, 2003; Rodehutscord and
Pfeffer, 1999; Booth and Allan, 2003; Lupatsch and Kissil, 2005; Peres
and Oliva-Teles, 2005; Glencross et al., 2007, 2008; Hatlen et al., 2007;
Glencross, 2008, 2009). This technique has also been applied to
digestible protein (DP). The maintenance requirement for DP (DPm)

/ Aquaculture 305 (2010) 150–158 and the efficiency of utilization of DP for gain above maintenance (kDPg) have been determined in red drum.4 17. Norway) (Table 1). Atlantic cod and barramundi (Lupatsch et al.3 °C (mean ±S. It is not a prerequisite with this method that the nutrient or energy is deficient in the diet. Glencross. Sweden). gilthead seabream. Hauler et al. In addition. wholemeal flour Vitamin mixa Mineral mixa Yttrium oxide Chemical composition Dry matter (DM) In DM N × 6. The use of an Nfactor of 6. Yttrium oxide was added to the diet as an inert marker for digestibility determinations. Hauler and Carter. Thus.. (2007) determined that ration level did not affect the efficiency of utilization for gain or the maintenance requirement of the deficient AA.0 18. rainbow trout. more specific knowledge about the requirements of the individual AA is important for formulating diets adequately.1. we use the term Requirements by Ration Level (RRL) to better describe this method. after hydrolysis in 6 N HCl for 23 h at 110 °C. Helland et al. d Asx represents Asp and Asn (analyzed as Asp) and Glx represents Glu and Gln (analyzed as Glu). 1997. 1998. Abboudi et al. 2005. Diet A fishmeal-based extruded diet (4-mm pellets) was produced by Nofima Ingrediens (Fyllingsdalen. When comparing the weight of the SumAA and that of CP. European sea bass. 2003. 2006). 1998. 2008.c Crude lipid Starch Ash Energy Indispensable amino acidsb Arg His Ile Leu Lys Met Phe Thr Trp Val Dispensable amino acidsb Ala Asxd Cys Glxd Gly Pro Ser Tyr 686 213 87 10 4 0. 1972). Ohta and Watanabe. 1998.. the weight of the anhydrous. (2004). The requirements for maintenance and for gain above maintenance for individual AA in fish have been determined using diets with gradient levels of a deficient AA (Rodehutscord et al. Booth and Allan.4 13. This process results in an increase in the weight of each AA because of the water molecule that binds with it. 2008. 2006. Protein-bound amino acid form. as has been done for DP and DE. in line with the 4. overnight).0 10.. Materials and methods 2. IL. 2006. Crude protein (CP) consists of both amino acids (AA) and nonprotein nitrogenous components. 2008.25 in diets using fish meal as the protein source overestimates the true protein content by approximately 30%. Argent Chemical Laboratories. The use of free AA in the calculation of the SumAA overestimates the protein in the sample because of the water molecules. Because of the confusion associated with the use of this name also for a statistical design. McGoogan and Gatlin. c The sum of those amino acids that can be synthesized into protein calculated using the weights of the protein-bound forms of the amino acids.25 Sum amino acidsb. 2007. Bodin et al. Normally. until constant weight). The Norse LT-94 151 Table 1 Formulation and composition of the experimental diet (g or MJ kg− 1). Growth trial Three weeks before the start of the trial. Because the requirement for this nutrient is influenced by both components it does not necessarily represent the animal's metabolic AA requirements for growth. Formulation Fish meal (Norse LT-94) Fish oil (NorSeaOil) Wheat. (1987) estimated the N-factor for fish meal to be 4. Ireland). 500-L cylindro-conical tanks (0. supplied with seawater (33.1 33. 2. USA). Glencross et al. 2009). The AA in the diet were analyzed using a Biochrom 30 AA analyser (Cambridge. USA) and yttrium (inductively coupled plasma mass spectroscopy at Bioforsk. WA. Peres and Oliva-Teles. The biomass in each tank was . The diet was analysed for dry matter (DM) (105 °C.6 25. Norway). U.8 60. 12 l min− 1) with a temperature of 9.0 305. 2. The “factoring” of the requirements with this technique is probably the basis for the name that has often been used to describe it — the factorial design. Bodin et al.8 94. Höganäs. and provide more information about the use of CP or SumAA for expressing the protein quality of a diet. Kjeltec Auto System. crude lipid after HCl hydrolysis (Soxtec HT6.7 22. 2007.1 940. ash (550 °C. Tryptophan and tyrosine were analyzed after basic hydrolysis (Hugli and Moore.9 6. Sweden). 2008.3 a As described by Mundheim et al.9 ppt salinity..1 46.. 2007. DE. has the maintenance requirement or the efficiency of utilization for gain been determined for the AA in a single diet fed at different ration levels.75-m diameter). silver perch.7 3. In none of these works however. the fish had been in seawater for 54 days. The N-factor is established by dividing the SumAA by the nitrogen concentration. starch (determined as glucose using the “GODPOD method” after hydrolysis by α-amylase and amylo-glucosidase. Thus.) following the EC Commission Directive 98/64/EC (1999). 2009). The intention of this experiment was to establish the DP. b fish meal was made from 65% herring and 35% herring waste (Aas et al.25. Boisen et al.S.9 17. it is important to use the correct N-factor for calculating CP..8 367. Höganäs. white grouper. Ås.J. Lupatsch and Kissil. protein-bound form of the AA must be used in the calculation of the SumAA.. Bray.4 18.7 18. Atlantic salmon (Salmo salar) were placed in 16. and then the weight and length of each fish was measured. 2007. Rollin et al. which is defined as the nitrogen concentration of the sample multiplied by an N-factor. Redmond.. the method was expanded to include determination of the requirements for maintenance and for gain above maintenance for the indispensible AA (IAA).8. After a 2-d fast. The sum of the AA (SumAA) that can be synthesized into protein gives a more accurate picture of the protein in a sample than the measurement of CP.7 25.8 541.7 35. Hauler et al. the weight of the free (hydrated) form of the AA is used for expression of the AA in the nutritional literature.9 ±0.2. 2005.9 for fish meal calculated from the data of Aas et al. Moline. The analysis of AA starts with acidic or basic hydrolysis of the protein.. crude protein (N × 6.D. Parr Instrument Company.). Hatlen et al..1 13. the fish were anaesthetized with tricaine methane sulfonate (MS 222. Megazyme. Tecator.1 32. energy (Parr 1271 adiabatic bomb calorimeter.K. Tecator.8 ± 1. the requirements simply characterize how the consumed and/or digestible nutrient or energy from the diet is utilized in the body. and ME requirements of Atlantic salmon post-smolts for maintenance and the efficiency of utilization for gain above maintenance using increasing ration levels.4 40. At the start of the trial.1 26. Peres and OlivaTeles. 2001. (2006).

killed with a blow to the head and sampled for whole-body analysis. Forsberg. AA weights in the diet and whole body are in the protein-bound form. The other groups were fed for 51 days. European sea bass and white grouper (Lupatsch et al. respectively (Elliott and Davison. Percentage data were subjected to arcsin square root transformation before statistical analysis. Retention of N. Following the initial 2 weeks of the trial. Three groups of fish were left unfed for 28 days and then terminated.. The oxygen consumption data for each tank was averaged within each day and then averaged over the experimental period. Water temperature and salinity were measured daily. HIME and HIheat represent the HI for gain (HIg). Therefore.4. from eight fish per tank. whereas. At the end of the trial.. The tanks were equipped for semi-open. All AA are expressed in the protein-bound form. ash. Lupatsch and Kissil. The efficiency of utilization of DP. The tanks were checked daily for dead fish. Heat increment (HI) was calculated using two methods: 1) from the comparative slaughter data. 2.. weights were taken of the whole body. The rations were adjusted based on intake levels the last 3–4 days.8. DE and ME were estimated as the xintercepts in the linear regressions (y = 0). 1981). The samples were stored at −20 °C until being analysed for DM. HIheat = heat − maintenance. Heat (energy used for maintenance. energy. 2001a. 1995) and the oxygen concentration was measured with OxyGuard® probes (OxyGuard International A/S.152 S. and approximate feed intake for each tank was calculated by taking into account the waste feed level and the percentage recovery of dry matter from the diet in the system (Helland et al. and then the weight and length of each fish was measured. Statistics The data were analyzed by one-way analysis of variance and means were compared using the least-squares means procedure (SAS software.7 ±10. Cui and Liu. Denmark). which included the heart and kidney. All fish were sampled at the end of the trial. Apparent digestibility (ADC).1 kg and the mean weight of the fish was 95. 2. Condition factor: 100 × (BW. triplicate groups were fed at levels approximating 25. Elliott and Davison.76 used to characterize a variety of salmonid species. / Aquaculture 305 (2010) 150–158 reduced to 3. assuming 15% of the nitrogen is excreted as urea and 85% as ammonia (Wright et al. with the following exception: the lipid analysis was done without HCl hydrolysis. The water inlet and outlet of each tank were closed for 10 min every 2. NC.1. Dressing percentage. CP (N × 6.. energy and AA as described above. The data were checked for homogeneity of variance using Bartlett's test. the rest of the viscera. 1998. fasted for 3 days to fully empty the gastrointestinal tract. 1975). respectively. Lupatsch et al. the non-faecal nitrogen excretion was calculated as the difference between digestible nitrogen intake and nitrogen deposition. For the determination of the requirements for maintenance and efficiency of utilization for growth above maintenance. Cary. 32 fish per tank). Helland et al. and the carcass. % body weight (BW) per day: 100 × (dry feed intake (FI)) × [(BW0 + BW1) / 2]− 1 × (days fed)− 1. In both of these calculations. molecular weight protein-bound form/free form = 128. 1995.3. Using oxygen consumption of 0. Specific growth rate (SGR). version 9. semi-closed respirometry (Grisdale-Helland and Helland. where C0 and C1 are initial and final concentrations in whole body. Maintenance requirements for DP.5 h and the water was recycled. HIME = MEi − maintenance − energy gain and 2) by combining the comparative slaughter and longterm respiration data. %: 100 × carcass weight × BW− 1.D. Birkerød. Apparent digestibility coefficients of the dietary nutrients and energy were calculated as described by Maynard and Loosli (1969). The faeces were freeze-dried and analysed for lipid. where BW0 and BW1 are initial and final mean body weights per tank. USA). Four tanks of fish were fed in excess to estimate maximum intake. The experiment was carried out with permission from the Norwegian Animal Research Authority. For lysine.1 g Atlantic salmon. 1956) determined the metabolic scaling coefficient for energy to be 0. 1/3 −1 . 1998. Welch's test for . A scaling coefficient for protein and AA has not been determined for Atlantic salmon.4 g (mean ± S. where subscript i stands for indicator and n for nutrient. When the variances were not assumed to be equal. The faeces were collected and frozen several times per day. % per day: 100 × (ln BW1 − ln BW0) × (days fed)− 1. starch. Sampling At the start of the trial. AA and yttrium as described above for the diet. %: 100 − (100 × {% feedi / % faecesi} × {% faecesn / % feedn}). The waste feed was collected daily. The tanks were equipped with individual lights and were designed to accommodate collection of waste feed from the effluent water in wire mesh boxes..19 = 0. for example. respectively. g) × (fork length. and a coefficient of 0. lipid. 2. The individual tank was the experimental unit. 1988.6 kJ per g O2 consumed. faeces were sieved from the effluent water using automatic collectors (Choubert et al. Feed efficiency ratio (FER): wet BW gain × FI− 1.. %: 100 × [(BW1 × C1) − (BW0 × C0)] × (Cdiet × FI)− 1. 2005). 10 fish were anaesthetized.25). as reviewed by Winberg (1956).9 and 23.. 1996). The biological oxygen demand was determined in each tank after removal of the fish at the end of the trial and was found to be negligible. CP (N × 6.81. For the energy budget. Hepatosomatic index (HSI): 100 × liver weight × BW− 1. the waste feed was analyzed for DM content and actual feed intake for each tank was recalculated. IAA. n = 512.7 for the geometric body weights was chosen for these nutrients based on determinations in gilthead seabream. the digestible IAA. The oxygen saturation level for all tanks was maintained over 85% with oxygen supplementation. and the energy loss associated with this excretion was calculated using enthalpy of combustion values of 24. 1975). 1997). Ivlev and Ivlev (1952. The sum of AA (SumAA): sum of weights of protein-bound form of AA that can be used in protein synthesis.1 kJ g− 1 nitrogen for ammonia and urea. The similarity between these coefficients and those determined with other fish species using growth data (reviews by Brett and Groves.5. the decline in oxygen concentration was measured in order to calculate oxygen consumption. Geometric mean body weight: (BW0 × BW1)1/2. and Cdiet is the concentration in the diets.J. cited by Winberg. AA and energy. the geometric body weights of the fish were scaled by metabolic weight exponents. maintenance includes activity and the HI at maintenance (HIm). After the trial. The fish were fed eight times per day using automatic band-feeders. Calculations Relative feed intake (RFI). 2001a. 1979 and Hepher. activity and heat increment) was calculated based on oxygen consumption (13. The remaining fish were killed for whole-body analysis. liver. 1982). Protein-bound AA: the molecular weight of the anhydrous AA residues corresponding to those in the protein chain.877. This was within the variation reported for the energy coefficient of 0. During the closed periods. cm)− 3. 1990.17/ 146. 50 and 75% of the intake of the full-fed groups. Thermal growth coefficient (TGC): 1000 × (BW1/3 1 − BW0 ) × (∑T) where ∑T is the sum day-degrees Celcius (Iwama and Tautz. SAS Institute. DE and ME for growth were calculated from the slope of the regression of gain of these nutrients or energy on the intake of these components. Inc.25). ash.87 to 40. 2003) led us to scale the weights of the fish to a metabolic weight exponent for energy of 0.

1b 94.10 ± 0.64 0.94 0.9a 1.05).1 ± 0.7 ± 0.1a 96.01 0.53 ± 0.01a 92. hepatosomatic index.05).1 ± 1.1a 96.5 ± 0.01 − 0. 3.10) compared to the fish fed the other ration levels (1.1b 94.20 ± 0.16 ± 0.04a 1.0 ± 0.05.00 0.4 ± 0.1a 95.5 ± 0. CA. quadratic decreases in protein.29 ± 0.00 1.2b 94.10 ± 0.1 ± 0.0d 93.1 ± 0.2a 95.15 1.8 ± 0.M.2ab 95.3 ± 0.1c 94.1a 97.8 ± 0.0 ± 0. Lys and Trp. / Aquaculture 305 (2010) 150–158 153 Table 2 Growth.0a 95.81 0.00 ± 0.01 3 3 3 3 4 95.0c 93.34 ± 0.9 ± 0.93 0.8 ± 0.4 ± 0.3 ± 0.3 ± 3. which decreased linearly.8 ± 0.b.4 154. ¶ Asx represents Asp and Asn (analyzed as Asp) and Glx represents Glu and Gln (analyzed as Glu). Relative feed intake level (% body weight d− 1) Dry matter Protein Sum amino acids Lipid Starch Energy Ala Arg Asx¶ Cys Glx¶ Gly His Ile Leu Lys Met Phe Pro Ser Thr Trp Tyr Val a.5 ± 0.7 ± 0.98 ± 0.4 ± 0.73 0.34 ± 0. differences among group means was performed.97 0. and are presented as means ± S.). FER. Leu.4 ± 0.5 ± 0.4 ± 0.4 ± 0. feed efficiency and organ indices of the salmon fasted for 28 days or fed the experimental diet for 51 days (mean ± S.00 0.3 ± 0. dressing percentage.6d 0. and a linear decrease in starch digestibility (P b 0.5 ± 0.73 ± 0.8 ± 0.E.67 0.21 ± 0.1a 96.5 91.E.5 ± 0.02a 3. GraphPad Software.0d 95.1a 94.1ab 5.0 ± 1.00d 94. Digestibility There was no mortality during the experiment.). 3.77 ± 0. Linear and quadratic effects of RFI on the dependent variables were also determined.1b 95.1c 6.3b 93.2c 93.3 ± 0.1 ± 0.7 ± 0.3 ± 0.6 ± 0. HSI.1c 94.7 ± 0.05 ± 0.7 ± 0.05) (Table 3).2 ± 0.1b 95.9 ± 0.89 0.. Dressing.0b 96.1b 94. respectively (Table 2). TGC.6 ± 0.3 ± 1.1 ± 0.7 ± 0.4 ± 0.0a 94.2c 93.01b 96.1b 85.3 ± 0.0c 96.1 ± 0.01b 1.0c 96.3b 94. § .c.5 ± 0.00a 92.0c 95.E.25d 1.00 ± 0.005c 1.1d 89.1b 94.1b 95.4 ± 0. The decreases in Table 3 Apparent digestibility of the experimental diet fed to Atlantic salmon at different ration levels (mean ± S.1a 95.05 for this term.95 0. The increase in feed intake level resulted in a parabolic effect on DM digestibility.98% BW d− 1 and the SGR and TGC varied from 0.78 0.05 107. San Diego.0a 95.86 0.7 ± 0.78 0.95 0.0a 86.8 ± 0.01 1. The feed intake level accounted for 100% of the variation in SGR and TGC. relative weight of gastrointestinal tract. Relative feed intake level (% body weight d− 1) n Body weight (g) Initial Final SGR (% d− 1) TGC FER Dressing (%) GSI (%) HSI (%) CF¶ Fasted 0.1c 93.05 level.5 ± 0.7a 97.5 ± 0.1c 94.86%d− 1 and 0.3a 96.10b 93.88 0.1a 94.01a 1. FER was significantly lower in the fish consuming feed at 0.7 ± 0.3 ± 0.90 0.9 ± 0.26 ± 0.2ab 95.1c 93.1a 96. Growth and feed efficiency RFI varied between 0.7 ± 0.6 ± 0.98 84.0d 97.1c 96.9 ± 0.2 ± 0.20 and 0.4 ± 0.18 ± 0.1b 94.1a 91.20 0.3 ± 0.1c Linear Quadratic 0.0b 94.1a 97. GSI.3 ± 0.1d 94.0b 84. CF.02a 93.8 ± 0.01 95.85 ± 0.3 ± 0.1a 95.84 0.18 ± 0.5 ± 0.1b 6.87 0.7 ± 1.89 0.3 ± 0.6 ± 0.6 ± 0.13 ± 0.2 ± 0.04d 1.D.01% d− 1 and TGC of −0. R2 is given for the quadratic function if P b 0. Results 3.1c 98.00 Linear regression.1b 95.3b 96.1c 91.01.1a 85.8 ± 0.41 ± 0.1 ± 0.06 (mean ± S.1c 82.2a 96.98 SGR.93 0.3 ± 0.1b 96.1a 96.7 ± 0.0 249. R2 is given if P b 0.92 Values with the same letter are not significantly different (P ≥ 0.3 ± 1.1b 85.1a 94.1c 94. Helland et al.1 ± 0.S.4a 5.3 ± 0.10 ± 0.3 ± 0.8 ± 0. feed efficiency ratio.2 ± 0. thermal growth coefficient. USA).20 ± 0.60 ± 0.1c 94.4 ± 1.3 ± 0.1b 98.1b 97.1 ± 0.0a 96. Test of linear and quadratic regression using relative feed intake level (not including fasted fish) as the independent variable.79).0a 95.21 to 1. condition factor.b. N = 512).00 0. The choice of using quadratic or linear regression was done using Akaike's Information Criteria (GraphPad Prism v.9 ± 0.37 ± 0.5 ± 0. lipid and energy digestibility. § Test of linear regression using relative feed intake level (not including fasted fish) as the independent variable.94 0.M.94 0.77–1.1b 95.7 ± 0.08 1.98 0.01b 2.34 to 3.86 ± 0. The salmon that were fasted for 4 weeks had a small decrease in weight and had an SGR of −0. R2§ 0.04 ± 0.6 −0.0c 94.53 0.0 ± 0.1 ± 0.2.c.1a 98.9 ± 0.0d 93.79 ± 0.1c 95.1a 94. a.2 ± 0. specific growth rate.3 ± 0.73 ± 0.83 0.0d 96.2 ± 0.0 ± 0.20% BW d− 1 (1.7 ± 0.78b 1.3b 94.J.1 ± 0.1.8 ± 0.6 ± 0.3 ± 0.d Values with the same letter are not significantly different (P ≥ 0.41.1a 94.8 ± 0.1a 95.02c 0.13± 0.1b 96.2c 93.1a 94.94 ± 0.1b 91.1b 95.0 ± 0.5 193.1a 96.00 ± 0.12 ± 0.01c 1. His.9 ± 0.20 ± 0. ¶ Initial condition factor 1.10a 1.d Regression R2§ 0.78 ± 0.02d 0.8 ± 0.1 ± 0.6 ± 0. 5 for Windows.97 0.1 ± 0.04 1.1b 97.01c 96.1a 95.1b 94. Differences between treatments were considered significant at the P b 0.94 0.1 ± 0.05b 1.4 ± 0.88 0.5b 94.M.7 ± 0.95 0.1d 93.4 ± 0.2a 88.98 1.1b 96.06c 1.2c 95. The increase in feed intake level also led to quadratic decreases in digestibility of the SumAA and all individual AA except Glx.0b 95.

03 4.0 17.1 16.c.89 ± 0.02 3. Met.06 5.95 ± 0.32 ± 0.01ab 4.7 d− 1 for Met and Trp to 20.15 ± 0. quadratic increases in the retention of the other digestible IAA were shown with increased intake (Table 6).63 ± 0.04 7.01ab 14.05b 28.01ab 4.08c Linear d 29.03b 4.04 10. / Aquaculture 305 (2010) 150–158 Table 4 Body composition of the salmon (g 100 g− 1 or kJ g− 1) (mean ± S.72 ± 0.08 6.36 0.03 10. The maintenance requirements for the individual digestible AA varied from 4.02 6.8 ± 0.06 5.M. Thr.08 4.01 8.c. Linear and quadratic regression tested using relative feed intake level (not including fasted fish) as the independent variable.96 ± 0. 3.0 mg kg− 0.06bc 2. R2 is given for the quadratic function if P b 0.01a 4.1b 2.2 12.83 ± 0. His.93 ± 0. The proportions of Ala.03 ± 0.10 0.40 ± 0.58 ± 0.05 ± 0.28 ± 0.5 mg kg− 0.02a 3.04 4.06 4.08 6.09bc 3.5 ± 0.01b 4. Helland et al.06a 2.12 6.57 ± 0.56 ± 0.13 ± 0. The maintenance requirement for DP was 117.3 ± 0.98 ± 0. Protein-bound form of amino acids.05 ± 0.01 3.21 ± 0.9 ± 0.02 6.03 9.9 ± 0.05ab 3.32 ± 0.4 ± 0. ¶ Asx represents Asp and Asn (analyzed as Asp) and Glx represents Glu and Gln (analyzed as Glu).59 ± 0.52 ± 0.98 c 27.73 ± 0.05 4.04 0.96 ± 0.). Trp and Val were heterogeneous and no significant differences were found among the treatments using the Welch test.00 in groups of fish consuming the least feed to 1.02 ± 0.82 0.21 ± 0.01 in those consuming the most feed (Table 2).85 ± 0.02 6.1 12.44 ± 0.03a 4.05 8.07d Quadratic 0.00 10.45 for Trp to 0.03 ± 0.02c 3.38 4.36 0.03 6.02 4. linear increases in the concentrations of lipid and energy and a linear decrease in ash concentration (Table 4).06 3.1d 2.1c 2.18 ± 0.73 ± 0.7 d− 1 for Leu.0 ± 0.18 ± 0. His.3 16.6 ± 0.1 ± 0.03 8.d Regression R2§ Initial Fasted 0.02 1.03 0.09 ± 0.04 4.1 16.03 10.04 ± 0.02b 2. § digestibility with increasing feed intake level were less than 2.3 mg kg− 0.8 ± 0.3.87 ± 0.1 2. Ile and Phe in the fish fed the lowest ration level was significantly lower than in fish fed at the higher ration levels (Table 6).64 0.30 ± 0.84 10.5% for all nutrients except starch.89 ± 0.30 6.05 4.01c 14. ¶ Protein-bound form of amino acids.05 4. AA and energy A quadratic increase in the retention of DP was found in the salmon with the increase in feed intake (Table 6).27 ± 0.E.06 4.05).29 ± 0.04 4.42 ± 0.13 ± 0.68 ± 0.02 0. The efficiency of utilization of DP for gain was 0.48 0.2 17.15 ± 0.10 ± 0.80 0. Except for digestible Met which showed a linear increase in retention with the increase in feed intake.2 ± 0.98 0.14 3.46 0.01 7.1 12.0a 6. Glx and Gly in the SumAA in the body decreased linearly in response to the increase in feed intake.13 ± 0.05 for this term.01 14.08 ± 0.97 ± 0.b.21 6.70 4.03 7.0 ± 0. whereas. HSI (1.68 ± 0.55 ± 0.32 ± 0.7%. R2 is given for the quadratic function if P b 0.01 ± 0.3 ± 0.0 9.19 4.b.07 7. The increase in feed intake resulted in a quadratic increase in the DM concentration in the whole body.05 4.15 ± 0.08 6.93 3.03a Regression R2§ 0. Test of linear and quadratic regression using relative feed intake level (not including fasted fish) as the independent variable.3 ± 0.99 ± 0.03 7. where the drop was 5. Leu. # § . The efficiency of utilization of the individual digestible AA for gain varied from 0.38 Values with the same letter are not significantly different (P ≥ 0.01 1.90 ± 0. 1 and Table 7).0 17.9 ± 0.41 Quadratic 0.01 3.16 ± 0.45 ± 0. Arg.98 Linear 6.53 0.04 ± 0. A quadratic decrease in the proportion of Met in the SumAA was also found when feed intake level increased.47 ± 0.20 0.14 9.2 12.45 4.154 S.02 (Fig.32 ± 0.73 0.05).0bc 7. Utilization of protein.06 ± 0.05 for this term.90 ± 0.9 ± 0.3 10.8 ± 0.2 ± 0.2 ± 0.41 ± 0.95 6.13 6.84 ± 0. Lys and Thr increased linearly (Table 5).02 8.1ab 7.53 ± 0. Relative feed intake level (% body weight d− 1) Initial Dry matter Crude protein Sum amino acids¶ Crude lipid Ash Energy 28.71 6. As with DP.03 4.01 3.02 1.63 1.06 ± 0.03 8.53 ± 0.03 0.09 ± 0.18 ± 0.0 6.26 4.51 ± 0.06 ± 0.11 ± 0.61 ± 0. Relative feed intake level (% body weight d− 1) Ala Arg Asx¶ Cys Glx¶ Gly His Ile Leu Lys Met Phe Pro Ser Thr Trp Tyr Val a. The concentrations of CP and SumAA were not significantly affected by feed intake level.98 14.01a 14. a quadratic increase in the retention of DE with increasing feed intake was also found (Table 6).09 4. Body indices and composition Dressing percentage decreased linearly and the relative weight of the gastrointestinal tract increased linearly with increasing feed intake level (Table 2).9 ± 0.02c 2.09 ± 0.d Values with the same letter are not significantly different (P ≥ 0.4 ± 0.M.91 ± 0. The DE and ME requirements for maintenance Table 5 Amino acid# composition of the salmon (% sum of amino acids) (mean ± S.51 Fasted 0.9 mg kg− 0.01bc 14.01a 4.53 ± 0.01 and of digestible SumAA 0.01 10.04%) was not significantly affected by feed intake level.62 ± 0.22 0.14 ± 0.20 25.64 ± 0.43 ± 0. The variance in retentions of DP and digestible Arg.7 9.61 ± 0.29 ± 0.6 ± 0.97 a.2 7.53 a 0.7 d− 1 (Table 7).6 12.62 ± 0.1a 2.06 4.02 1.17 ± 0.01 6.65 for His (Table 7). Lys.).03 ± 0. The increase in feed intake resulted in a linear increase in condition factor from 1.57 ± 0.44 ± 0.77 2.13 ± 0.01 4.0 2.4. the increase was steeper. the proportions of Cys.2 8. 3.93 ± 0.1 ± 0.03b 4.81 ± 0.1 12.2 6.03 5.03 6.03 4.4 ± 0.47 8.44 ± 0.02 1.0c 8.02 1. but in this case.26 ± 0.J.01 4.01 4.98 0.1 7.7 d− 1 and for the digestible AA 204.02 3.06 ± 0.63 ± 0.35 ± 0.07 4.73 b 25.69 0. The retention of digestible SumAA.E.7 ± 0.02 1.06 2.18 ± 0.04 ± 0.14 8.74 ± 0.20 ± 0.35 ± 0.

1 ± 1. oxygen consumption increased with each increase in MEi.2 Histidine − 4.7 41.82(±0. The energy balance was poorest for fish fed the lowest ration level.62 ± 0.7 ± 1.98 13.4 Threonine − 7.51 Leucine −12.5 5.9 ± 1.b.0 ± 1.2 55.4 ± 0.7 61.22(± 0.9 62. 4.M.J.3 DE −25.2 29.69 0.4a 51. At the highest ration level.1 57. The dashed line.3 132.3 ± 0.80 ± 0.8% of MEi.5 ± 9.98 Linear 41.3 56.3 Valine − 6.1 ± 8.6 ± 2. The solid line is derived from the fed groups: y = 112.91)+0.00 30.0 55.6 kJ kg− 0.1 ± 0.2 21. Fig.4 ± 35.05).6 ± 0. and the efficiency of utilization of DE and ME for growth were 0.98 7.01 and 0.9 ± 0.01 0.4 42.00. These groups were overfed at a level of 8. Linear and quadratic regression tested using relative feed intake level as the independent variable. the Fig.2 2.64(±0.5 ± 1. P b 0.0 2.3 0.0 ± 2. 2.2a 50. R2 =1.4 mg kg− 1 h− 1) (P ≥ 0.9(± 4.5 ± 0.99 204. In the other groups.58 ± 0.8 d− 1) (the fasted fish were included in the regressions).01 0.05).8 ± 0.99 11.02 0.0 ± 3.79 0.001.8b 26.7 d− 1) = −75.60 ± 0. Daily energy and protein gain per unit metabolic weight of Atlantic salmon fed increasing levels of metabolizable energy (ME) and digestible crude protein (DP).1 ± 1.9 0.9 ± 8.1a 56.1 14.8 60.7 56. Discussion The growth rates of the full-fed fish were high and 100% of the variation in growth was explained by the measured feed intake of the fish.0d 62.2 ± 22.7 0.7 d− 1 or kJ kg− 0.02 0.98 18.3 ± 1.0 0.4 26. Pb 0.9a 57. the sum of amino acids¶ (SumAA).02 0.4 ± 9.8 d− 1)=−25. R2 is given for the quadratic function if P b 0.7 ± 0. the indispensable amino acids¶ and energy (%. P b 0.3 0.5 ± 3. For the fed groups of fish.73 0.99 20. is derived from all groups of fish: y = 110.6 mg kg− 1 h− 1) and the groups consuming feed at the lowest level 155 Table 7 Regression coefficients (± standard error) in the equation y = a + bx describing the deposition of protein. For both the fed groups of fish and the fed and fasted groups together.5 ± 0.8 64.3 45.1 ± 0.0 ± 0.5b 23.95.1 59.4 49.8 d− 1).0 0.3 ± 0.02 0.53 or 0.65 0.4 ± 1.13)x + 0.6 ± 2.1 ± 11.53 0.49 ± 0.5 and 30.02 0.3 ± 0.1 60.0 ± 1.4 ± 3.75 0.9 Isoleucine − 7.79 0.22)x + 0.7 d− 1).3 ± 3.9 ± 2.64 ± 0.53 ± 0.7 ± 0.4 ± 1.02 0.001(± 0.6%. The energy budget for each ration level is presented in Table 8. respectively (Fig.57 ± 0.0 ± 0.74 0.8 5.25(± 0.8 d− 1.3 ± 0.0b 22.1 ± 6. a Protein −75.7 0.2 63.0 ± 2.7 55.45 ± 0.002)x2.1 Arginine − 6.73 0.94.1 ± 2.58 ± 0.2 ± 1.62 ± 0. R2 = 0.73% BW d− 1 where there was a strong trend towards a difference (P = 0.7 ± 2.1 ± 2.80 ± 0.8 0.S.1 ± 0.9 8.13(±0.01) × DP intake (mg kg− 0.7 ± 2.10) + 0.3 ± 3.4 26.0 ± 1. respectively.5 ± 11.4 ± 2.01)×ME intake (kJ kg− 0.5 ± 2.6 ± 6. At the lowest ration level.).02 0.84 0.3 37.0 61.9 ME −25.1 ± 0.2 0. mean ± S.E.82 ± 0.9 ± 0.5 ± 5. Analysis of variance showed no difference in oxygen consumption between the fasted groups (110.98 11.1 ± 0.1a 60.0(± 8. Helland et al.0a 59.01 0. except between the groups that consumed feed at 0.99 117.9 Phenylalanine − 4. 3 to 8% of the input was not accounted for.4 63.001.4 ± 3.2 64.05).8 ± 9.73) +0.4 Protein-bound form of amino acids.0 28.76 0.7 SumAA − 117.3 ± 28.1 8.7 d− 1 or kJ kg− 0. Protein SumAA Arg His Ile Leu Lys Met Phe Thr Trp Val Energy Relative feed intake level (% body weight d− 1) Regression R2§ 0.59 ± 0.2 12.3 ± 1. Protein-bound form of amino acids.1 ± 1.1 ± 9.00 31.9% of MEi.2 ± 0.2 ± 1.1 58.7 d− 1 or kJ kg− 0.98 a.02 0.2 − 6. the individual indispensable amino acidsa and energy as functions of intake of the digestible nutrients. HIheat accounted for 11.8 57.20 0.0 7.45 0.5 ± 3. R2 = 0.6 Lysine −11. / Aquaculture 305 (2010) 150–158 Table 6 Retention of digestible protein.2 6. the sum of amino acidsa (SumAA). 2).3 ± 2.01 0.6 ± 3.9 58.6 52.4 ± 2.5 1. Routine oxygen consumption of Atlantic salmon as a function of metabolizable energy (ME) intake. maintenance requirement (− a/b) and loss of nutrients or energy during fasting.8 ± 3.5 60.0 ± 3. The fasted fish are included in these regressions.2b 60.7 59.9 32.99.0 25.99 4.65 ± 0.7 ± 1.4 ± 2.82 ± 0. (118.4c 61.99 10.9a Quadratic 0.7 ± 2.0002)x2. .001. 1. ¶ § were 31.3 60.8) + 0.5 0.c. with 13% of the input not being accounted for.98 4.4 19.05 for this term.5 ± 0.1 ± 2. Protein gain (mg kg− 0.5a 50.4 ± 8.001. whereas.83 0.0 30.01 Maintenance (mg kg− 0. whereas HIME accounted for 14. R2 = 0. digestible energy (DE) or metabolizable energy (ME) (mg kg− 0. P b 0. no HI for gain could be calculated.8 d− 1) 85.3 Tryptophan − 2.9 Methionine − 2.98 8.21(±22.d Values with the same letter are not significantly different (P ≥ 0.0 0.2 ± 8.5 ± 4. the fish lost energy in the body and therefore.6 Loss during fasting (mg kg− 0. which is partially hidden by the solid line.2a 57. quadratic increases in oxygen consumption were found with increasing MEi (Fig.02 0. Clear symbols represent feed-deprived groups and black symbols represent fed groups.01. 1 and Table 7).002(± 0.4 1.9a 57.9 a b (efficiency R2 of utilization) 0.3 ± 2.74 0.5 ± 11.8 d− 1) 0. Energy gain (kJ kg− 0.

Heat. 2008). Hatlen et al. metabolizable energy intake. 1998. Azevedo et al. the fasted groups were included. In the present experiment.48 kJ kg− 0. Glencross. All of these estimates greatly exceed the present value of 0. digestible energy intake. 1981) does not allow for other losses from MEi above maintenance than HIg (Eq.8 d− 1.5 13. HIME curve.5 160.1 24. MEi. 2003). the k-factors for HIg range from 0. HIheat is similarly curved in relation to MEi. especially when the relationship between MEi and HI is linear.6 ± 1. because the fish still had the opportunity to slip with the current since there was no netting across the radius of the tanks. indicating that the rate of loss of energy below maintenance (km) occurred at the same rate as the deposition of energy above maintenance (kg). When the negative values for HIheat are removed from the dataset. Additionally. expressed as CP or SumAA.17 for smolts found by Grisdale-Helland et al. Bureau et al.J.6 Undefined Undefined − 1.5 64.18 to 0.. 2005. Assuming that the comparisons are real.8 d− 1. A quadratic decrease in the Met concentration in the SumAA also occurred. Thus.5% BW d− 1 (1. as we had seen earlier in a similar experiment with Atlantic cod (Hatlen et al. Meyer-Burgdorff et al. (2001b) and Hatlen et al..2 ± 0..1 109. 8 d − 1 ) in the following manner: HIME = 0. Using Eq. There were few indications of fin biting. kHIg values can be calculated for Nile tilapia (0.3 76.1 ± 1. regardless of the energy substrate.1 36. also by definition. submitted for publication).2 38..3 7. The strong influence of ration level on HI is well documented (Brett and Groves. Atlantic cod. 1998. an extrapolation of the linear relationship between MEi and energy gain was not necessary because the 20% feed intake level resulted in an energy balance that was just below maintenance. MEm is MEi when energy gain is zero. 1998. HIME is zero at zero MEi. where HIg is undefined.7 ± 0.1 ± 2.8 ± 1. Above the lowest intake level. this resembles the situation seen when different dietary protein sources were fed to Atlantic halibut (Helland and Grisdale-Helland. Relative feed intake level (% body weight d− 1) GEi DEi MEi Heat HIheat HIME Energy gain 0. In addition... 2001b. was not influenced by the level of feeding.4 ± 1.8 d− 1.6 ± 0. Results with European seabass. 2006.8 ± 0. Low feed availability may have led to greater feedsearching behaviour. 2003). Thus. 1991.1 ± 1. This is similar to results for Atlantic cod (Hatlen et al.3 31. These changes in AA concentrations in SumAA indicate a shift in the type of protein that was synthesised.35) and rainbow trout (0. Booth and Allan. the relationship between MEi and HIheat is linear.96).46 to 0.10.30 to 0. His.. (1). Nile tilapia. The linear increase in condition factor with increasing feed intake was related to linear increases in whole-body fat and ash and a quadratic increase in DM.2 ± 1. 1989. In these relationships.2 26.. Bureau et al.1 ± 1.02 from 0.. gross energy intake. HI at maintenance thus becomes: HIm = 0. As noted earlier.3 0. where the efficiency of utilization of DE was greater between maintenance and fasting than above maintenance.. Lower kDEg values.18 for Atlantic salmon post-smolts and 0. The linearity in the relationships between MEi and other parts of the energy budget gives no indication that the HI should be different above or below MEm.156 S. but species differences are evident. ranging from 0.195X (R2 = 0. Lys and Thr concentrations increased linearly. / Aquaculture 305 (2010) 150–158 Table 8 Energy budget of salmon fed the experimental diet for 51 days (kJ kg− 0. This latter situation is one that is often observed with warm blooded animals (NRC.6 ± 0.83. where X is MEi. the HIME is −7.   kHIg = 1−kMEg = ð1−0:82Þ = 0:18 ð1Þ A traditional RRL between MEi and HIME (kJ kg− 0. HIME. submitted for publication).6 kJ g− 1). The concentrations of several AA in the protein changed however: Ala.78). however. Helland et al. 1981). GrisdaleHelland et al..4 78. (2007) were unable to show that kDEg in fish was influenced by the dietary energy level.20 0. similar to that found in rainbow trout (Storebakken et al. The energy used for locomotion by the salmon is included in the estimates for the maintenance requirements.7 and 0. it is logical to express HI M E (kJ kg − 0 . It is possible that the fish used different energy substrates dependent on the energy intake level.1 ± 0. (submitted for publication). The R2 for the linear relationships between both MEi and DEi and energy gain was 1.80. Azevedo et al.6 ± 0.8 ± 1. All of these measurements indicate that the fish performed well. Both HIheat and HIME values in the present work were strongly affected by feed intake.. and the origin must therefore be the starting point for the MEi vs. This is simply because HIheat was calculated by deducting a constant (maintenance) from heat. or it may have resulted in reduced activity in order for the fish to spare energy.39) (Meyer-Burgdorff et al. 2007. 2006).2 ± 1. Ohta and Watanabe. and the efficiency of utilization of energy for gain was increased by 0. 2005. Glx.8 ± 0. The influence of feed intake on HI is so strong that it questions comparisons between published values.15 and 0. the feeding rate may have affected the activity of the fish.37 to 0. Will RRL logic applied to this energetic term also create feed intake independent parameters? This should be possible because the energy scheme (NRC. Ohta and Watanabe. 1976.5 5.. Lupatsch et al. and were exposed to low levels of disturbance.1 ± 0.5 GEi.5% overfeeding.3 ± 0.1 102.4 172. This is because the water speed in the tanks was standardized. large barramundi and rainbow trout have shown kDEg values between 0. 2008) and white grouper (Lupatsch et al.48 + 0..6 = 5.1 112.73 0. HIheat. Glencross et al. whereas.2 27.5 kJ kg− 0. heat increment for gain from heat [Heat − maintenance (30.. 1998) and Atlantic salmon smolts (Grisdale-Helland et al.2 ± 2.8 d− 1) yields the equation HIME = − 7. gilthead sea bream..195 × 30. but in contrast to the situation with gilthead seabream (Lupatsch et al.6 kJ kg− 0. Another interesting aspect of this dataset is that we have compared long-term respiration ..8 82. Atlantic salmon have a low maintenance require- ment for energy (present work. HI cannot be negative. But. Arg. oxygen consumption (g O2 × 13. but this was not quantifiable.6 ± 0.9 23. This large difference in kHIg values may reflect both species and dietary differences.. The quadratic relationship between oxygen consumption (heat) and MEi may be the result of changes in the activity level of the fish over the range of intake levels. Glencross et al.00. have been reported in carp. at zero MEi. The maintenance estimate was reduced by 0. by definition. as more feed was consumed. and Gly concentrations were reduced linearly with increasing feed intake level. 2008. 2007. The protein concentration in the fish. 2006). Atlantic salmon post-smolts are very efficient in utilizing DE for growth (kDEg 0.22.9 units from 31.9 119. Grisdale-Helland et al.80). Cys.9 18. with a slope of 0. 2005. 2003. The coefficient used assumes constant efficiency of energy conversion. Another explanation for the curvature of these lines may be the use of a single oxycalorific coefficient at each ration level in converting oxygen consumption to energy. when MEi was used instead of DEi. other groups had less than 0.1 ± 2. (1)). 2006).9 ± 1.1 155. It could also reflect a systematic problem that obscures comparisons between the experiments. 1979). Therefore.8 d− 1). silver perch and white grouper (Huisman. it would be interesting to evaluate factors influencing kHIg. 2007.8 (Peres and OlivaTeles. Azevedo et al. Comparisons on a kHIg-basis eliminate this problem. as confirmed in smolts (kDEg 0. but fell at the lowest intake level to 1.97 kJ kg− 0. heat increment for gain from ME (MEi − maintenance − energy gain). Lupatsch et al. 1989.8 d− 1)].9 49. FER was very high in the fish fed at or over 0.4 ± 0. DEi. submitted for publication).53 0.22. although this may involve interacting factors. The energy consumed for swimming activity may not have been equal in each tank though. 1998.3 ± 0..98 29. 2007).195X.70.

The authors argued that the efficiency of utilization of digestible Lys over 100% could be explained by microbial synthesis of Lys in the gut. we observed a change in the distribution of significance if the IAA were expressed in the free or the anhydrous form. supports this contention. J. Y.. Lys and Met levels when the AA were expressed as percentages of the SumAA...L. E. W. B.. Our approach to calculate the requirements for each IAA has also allowed the determination of the requirements of SumAA that can be synthesised into protein.. Helland et al. 2008. 83. 1998. Blaxter. as confirmed in the trial of Grisdale-Helland et al.F.. C. This was explained by the presence of non-protein nitrogen in the CP. van Milgen. 477–485. In contrast. Camb. The difference in the maintenance requirement for DP and SumAA is an indication that the non-protein nitrogen has a much higher rate of recycling in the body than the AA. whereas. Bureau. D..S. 1998. le Boulenge. 1997). G. in a single trial. Booth and Allan. References Aas.. as shown in Table 6. Larondelle. Thus.. 227–238. Y.75 in rainbow trout fry fed wheat gluten based diets containing graded levels of crystalline Lys at 50 to 100% of the dietary Lys requirement. Mambrini. As shown in the early work of Lofgreen and Garrett (1968). Glencross.64 for the salmon.. submitted for publication).33–0. Knowledge about how the RRL-parameters for the individual IAA behave in a surplus situation. Cho. Grisdale-Helland et al. Knowledge about the effect of other biotic and abiotic factors on the RRL parameters will be important for standardizing the experimental protocol and the use of the RRL parameters in growth models. X. 1998. 120A. 369–383. M. T. 13. Ooghe.. the DPm of 117 mg kg− 0.) fry with the diet dilution procedure. it seems that the efficiency of utilization for gain above maintenance is a good inverse indicator of the dietary level of the AA with respect to the fish's minimum requirement.. (2007) reported that the kLys was 0. 1966. J. N. the kLys in maize gluten based diets or diets containing a sesame oil cake and wheat gluten mixture. T.. 67–75. Abboudi.25 does not help in this analysis. Aquaculture 274. 1998. 3 g red drum. Agric. where long-term comparative slaughter experiments normally yield lower estimates for maintenance and gain than respiration studies that are generally short term (NRC. 1983). European seabass. (submitted for publication) were wheat gluten and fish meal. D. Bodin. Booth and Allan.. S... Aquat.. 2005. 2007..55 (Lupatsch et al. 2005). Terjesen. and might camouflage significant differences between treatments. P. J. Similarly. Acknowledgements This experiment was financially supported by the Norwegian Research Council.. S. nutrient and energy utilization and waste outputs of rainbow trout (Oncorhynchus mykiss).A.. 357–368. X.82 reported for barramundi (Glencross.7 d− 1 for salmon is low compared to other species (300–860 mg kg− 0. The RRL system can be expanded into a diet evaluation system that utilizes the maintenance requirements and net efficiencies of utilization of the individual IAA and energy.P. Peres and Oliva-Teles. Atlantic salmon smolts.A. Sci. This is especially important for Lys since it is often the first-limiting IAA in feedstuffs and is also prone to processing damage (Maillard reaction) that is difficult to assess in chemical evaluation systems.5 by 4. but also the biological availability of the IAA affects the efficiency of utilization for gain.6–2. Aquacult. submitted for publication). Effects of diets containing a bacterial protein meal on growth and feed utilisation in rainbow trout (Oncorhynchus mykiss). These species are more efficient than gilthead seabream. Garrett and Johnson.. Helland. Sci.. Larondelle. McGoogan and Gatlin.E.. T. 2007.7 d− 1 for Atlantic cod. The much lower retention of digestible SumAA compared with DP at the lowest ration level. 2008. Leeson. Bureau. it is much lower than the value of 0. Azevedo. Helland and Grisdale-Helland (2006) studied the effect of replacing fish meal in the diet with wheat gluten on AA composition in Atlantic halibut. Lupatsch and Kissil. 1981... Rollin.06) between dietary treatments in the whole body His. Leeson. Hatlen et al. We found differences (P ≤ 0. Lied.2 times higher than the minimum requirement for postsmolts (Berge et al. F. (2007) showed that kDPg is reduced when the dietary CP level is increased. Abboudi.. Sveier. Anim.. Nang Thu et al. Threonine requirements for rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) at the fry stage are similar. Azevedo. Rollin.7 d− 1. W. the dietary Lys level was 1.J. silver perch and barramundi) (Lupatsch et al. In the present trial.. Wauters.. Bakke-McKellep. Berge.. at the minimum requirement point and in a deficient situation. 2006. Lupatsch et al.9 as the N-factor for the protein instead of 6. Abboudi.1 mg kg− 0. We suggest that the differences in the efficiency of utilization for gain above maintenance between the individual AA are related to their different metabolic fates and the dietary level of each AA in relation to their minimum requirement. Nutrition of Atlantic salmon (Salmo salar). 2009. Aquaculture 261. rainbow trout. more efficient absorption of the AA in wheat gluten because of denser and more solid digesta in the fish than may have occurred with the other protein sources and negative influences of imbalanced AA in the maize gluten and sesame oil cake. Grisdale-Helland et al. 1998). each feedstuff contributes differently to energy gain in the animal (termed NEg value of the feed). Hatlen et al. E. was 0. Therefore... Hatlen. 2006. . but no differences when they were related to the CP concentration in the body. Ooghe.. 1998.-B. Physiol. Ooghe. silver perch and white grouper that showed kDPg values of 0. because the DP maintenance estimate will actually be reduced from 117.S. Nutr. Determination of the threonine requirement for maintenance in Atlantic salmon (Salmo salar L. Utilization of the energy and protein of the same diet by cattle of different ages. Grisdale-Helland. 2007..7 d− 1(McGoogan and Gatlin.. 2007) and only 62% of the digestible Lys in the mainly fishmeal-based protein was used for gain above maintenance.... Espe et al. Larondelle. gilthead seabream. Wainman. the requirement and metabolic effect of lysine. Additionally. Biochem. Rollin. Glencross. Aquaculture 261. 67. Glencross et al. 353–365. As Lys is an AA with few other metabolic fates 157 than being incorporated into protein (reviewed by Flodin. (2001b) and Hatlen et al. The authors noted that possible reasons for this difference included slower. The use of 4. Together with the observations of the different requirements of CP and SumAA in the present trial.M. (submitted for publication).. can be used to evaluate the sufficiency of all of the IAA in a diet.P. 2008). Ohta and Watanabe. Peres and Oliva-Teles. 842–851. 2008. W. Y.. Except for results from an experiment with 5-g red drum that showed a DPm of about 150 mg kg− 0. M. A. 2009). J. J. Effects of feeding level and water temperature on growth. Comp... Protein and lysine requirements for maintenance and for tissue accretion in Atlantic salmon (Salmo salar) fry.25 to 92. This is in contrast to the results that are generally obtained with warm blooded animals. and supplemented with graded levels of crystalline Lys.63. 2005. DPm is a poor estimate of the “true protein” maintenance requirement.. European seabass and rainbow trout (Ohta and Watanabe. Mambrini.. these results indicate that the common practice of expressing the AA concentrations as free AA per unit CP is questionable. Comparing efficiency of metabolizable energy utilization by rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) using factorial and multivariate approaches. 1998. (submitted for publication) showed that the same situation occurs with individual AA. The main protein sources in the diets used in the trial of Grisdale-Helland et al. 2007.9/6. It is very common to express the AA concentrations in a sample in relation to the CP content.L.W. P. not only the dietary level. is similar to that reported for Atlantic cod.. H. 2003. Clapperton. 2007.J. S.. 1998. 281–290. K.Y. X. Living Resour. / Aquaculture 305 (2010) 150–158 results with long-term comparative slaughter results and obtained very similar values for the estimates of HI. T.. Glencross et al. the protein source in the diet seems to affect the efficiency of utilization of the IAA. 2005. B. The maintenance requirement for SumAA is 173% higher than that for DP. 1998. 11. The efficiency of utilization of digestible Lys in salmon smolts was reduced from 108% to 85% when the dietary level was increased from a deficiency to a level just over the requirement in a wheat gluten and fishmeal-based diet (Grisdale-Helland et al. The kDPg of 0. 2003. B. The technical staff at Nofima Marin is thanked for their care of the fish and for carrying out laboratory analyses.

Pfeffer.25 from total nitrogen to protein. M.. Response of rainbow trout (Oncorhynchus mykiss) to supplements of individual essential amino acids in a semi purified diet. Transl. Randall.. Helland. 1997.).J. Nutr.. 1166–1175.J. Energy and nitrogen partitioning in 250 g Atlantic cod (Gadus morhua L.. 2001a. Nutr. Corrent.A. Aquacult. Replacement of fish meal with wheat gluten in diets for Atlantic halibut (Hippoglossus hippoglossus): effect on whole-body amino acid concentrations. Clarias lazera (C. Sciaenops ocellatus. 32 (Suppl.C. Exp. Res. Biol.. Lupatsch. 2009. C. McGoogan. S. 175–189. Peres. J. E. Hauler. parr... J. B. A. Energy equivalents of oxygen consumption in animal energetics. Garrett. Plisetskaya... de Saeger. Agric.. E. Rollin.). 95–107. 1995. 317–326. K. 2001. 2008. 2008. D. 1–17. M.M. Aquaculture 261. Y. Lupatsch. O.. 1212–1221. Acta. 1988. Abboudi. 1998.J. Res. G. Fish. 1982. 6th edition. Günther. G. Sci. 2006. Davison. T. Y. Aquaculture 225. 218–229. Reduced water oxygen levels affect maximal feed intake. M. D. S. A. J. European sea bass (Dicentrarchus labrax) and white grouper (Epinephelus aeneus): energy expenditure for protein and lipid deposition. Gatlin III. Dods. Grisdale-Helland. Nutr. Aquaculture 273.. 97A. Carter. Growth.. Luquet. Food conversion efficiencies at maintenance and production levels for carp.. I. Protein level affects the relative lysine requirement of growing rainbow trout (Oncorhynchus mykiss) fry... Rate of metabolism and food requirements of fishes. Biochem. 2008.. Ooghe.O.O. R. 613 pp. Comparison of the lysine utilization efficiency in different plant protein sources supplemented with L-lysine·HCl in rainbow trout (Oncorhynchus mykiss) fry. Nang Thu.W.. Washington. Oltjen. Sklan.. A system for expressing net energy requirements and feed values for growing and finishing beef cattle. submitted for publication. Becker. Br. Peres. K. A. R. K. N... Boisen.. S... W.) fed different dietary proportions of vegetable protein sources in combination with two fish meal qualities. McCafferty.. 1975.W. Sci. 1363–1370. The minimum dietary lysine requirement. N. J. Dods..J.. 279–352. Aquaculture 270. Aquaculture 96. Aquaculture 248. 4. G. Lupatsch.. J. Comparison of energy budget among six teleosts — II.. T. 185–189.F.J. Can. I. 169–174. Chem. 2005. McGraw-Hill. 2828–2834. Detavernier. Aquacult. Lates calcarifer based on Australian production conditions... Helland.. pharmacology. S. Effect of feeding level on growth and nutrient deposition in rainbow trout (Oncorhynchus mykiss Walbaum) growing from 150 to 600 g. 29–41. B. Elliott.. NY.. 1–202. L. Maynard.. Rodehutscord. K.Y. Cambridge. Kissil. B.). Fish. Storebakken. Coll. 359–362.C.E.E. 64. and toxicology of lysine.N. A.. P. T.. Metabolic rates. T. X.K. B. In: Hoar. J. D. Growth variation and fin damage in Atlantic cod (Gadus morhua L. J.. Nerland. National Academy Press. Protein and energy metabolism of European seabass (Dicentrarchus labrax) juveniles and estimation of maintenance requirements. 1981. Hope. S. E. Hung. E.. & V. H. Aquacult. 95. Hua. D. Petri. Glencross. 1090–1098. Salmo salar L. Gatlin III. Tautz.. I. Effect of feed preparation methods on dietary energy budgets in carp and rainbow trout. Bodin. W. Animal Nutrition. Y. W. 1968. D. Nutrition of Pond Fishes. Nutrient partitioning in rainbow trout at different feeding rates. Am. 14..D. T. Biol... Cambridge University Press. 2007.. Meyer-Burgdorff. Osman. A factorial growth and feed utilization model for barramundi. P. 289–302. G. Aquacult. Carter. P. 1979.W. Hugli. Glencross. S. 123–129.. Academic Press. 1983. B. A. Digestibility in fish: improved device for the automatic collection of feces. Winberg... 2001b. Pfeffer..K.. S. El-Mowafi. 14. 793–806. 31. A. pp.S... E.C. Aquacult. Res..D. N. Anim. for protein and energy based on weight gain and body composition. Rodehutscord. A simple growth model for salmonids in hatcheries. 57. 1996..L. 157–163.. 37.G. 102. Nutr. 167–177. body composition and protein utilization in gilthead seabream (Sparus aurata L.. 2004. Ohta. 299–304.G.S.. Kissil.. Wauters.158 S. Helland. N..F.. S. 28.) given graded levels of feed with different protein and lipid content. . Parkouda. Bioenergetic relations of body weight and feeding level. 147–156..G.R. 360–373. 2009. J. Wathelet. 2006. The metabolic roles. W. D.. W. Physiol. Aquaculture 275. P. Cui. Cyprinus carpio L.. Lofgreen. Scand.M. Larondelle. 7–21.J. 71–80..M. S. Helland. Glencross.. 1976.J.. Aquacult. 191–203.. J. G. Calvert. 37. Choubert. Rutherford... 1981. Sklan. S. Oncorhynchus mykiss. Evaluation of the influence of drying process on the nutritional value of lupin protein concentrates when fed to rainbow trout (Oncorhynchus mykiss). 259–273.P... S. D. Kissil. 234–245. Lemme. 198. Salmo gairdneri Richardson. A. Grisdale-Helland...B. 2007. Nutr.. 1983. W. B. Grisdale-Helland.. 127–135. Watanabe. B. 477–488.. 11. J. USA. B. G.. Lupatsch. 38.R. Res.J. K. Liu. S. Energy metabolism in Oreochromis niloticus. 15. Aquaculture 35.A. Garrett. Comp. 1956. B. Ichthyol.. A. 1). Growth and production of the African catfish.. Hawkins.W. Maintenance threonine requirement and efficiency of its use for accretion of whole-body threonine and protein in Atlantic salmon (Salmo salar). Aquacult. C. Oliva-Teles. Ferrell. Anderson... Kissil. J.. 2006. Evans. J.W.. Optimization of feeding regimes for European sea bass Dicentrarchus labrax: a factorial approach. I. X. 127... Lupatsch. 129–138. and rainbow trout. Larondelle. B. J. C.. application.. J. Eggum.. Hauler.. Larondelle. C.M. Sipsas.. NRC. Comparison of energy and protein efficiency among three fish species gilthead sea bream (Sparus aurata). C. N. Sipsas. 2003.. Sklan... Aquat. Loosli. Sci. Lysine deposition responds linearly to marginal lysine intake in Atlantic salmon (Salmo salar L. Biochem. Metabolic requirements of red drum. B..) parr. 128. Utilization of digestible nitrogen and energy from four agricultural ingredients by juvenile silver perch Bidyanus bidyanus. Hogendoorn. Johnson. Anim. Nutr.. M.). B. Vol VIII. Grisdale-Helland.. 1969. 1997. Wright.. Oecologia (Berl. 9. Net energy systems for beef cattle — concepts. and future models.-H. J. Feeding regime does not influence lysine utilization by Atlantic salmon. Hepher. 83–95. Series No. Oliva-Teles. S. Rutherford.. 86.B. 1997. Y. Helland. Moore. Aquaculture 79. 283–291. Pfeffer.J. W. Felskie.. 23–31.. Sci. Brett. Aksnes. Pfeffer. 2003. 2007. Rollin. Aquaculture 263.I. T. 478–491. C... Kissil.. M. J.D. McCafferty. C. Espe. de la Noüe. 247. Grisdale-Helland. 1999. 27. B. Cho. Aquacult.. 649–656. G.M. 283–290. Groves.. 1–8. B... J. Aquaculture 179.W. Can. Aquaculture 272. M. 1990.. E.P.. Flodin. M. New York. E. 165–173. Determination of the tryptophan content of proteins by ion exchange chromatography of alkaline hydrolysates. Aquaculture 202... A.N. Forsberg. S. 1989.. D. Aquacult. G..D.. 195–201.. including an estimate of the maintenance requirement for several amino acids. Lysine requirement and efficiency of lysine utilization in turbot (Scophthalmus maximus) juveniles.) 19.. Rollin. Nutr.. III. 1998. Aquaculture 9. Sci. H. Appl.. H. (Eds. maintenance requirement and efficiency of lysine utilization for growth of Atlantic salmon smolts. Aquaculture 29.. S.A. 315–331.. Methods to assess optimal nutrient composition of diets for Atlantic salmon. J.. Fish Physiol.A.) fed at graded levels of feed restriction. Grisdale-Helland. A critical view on the conversion factor 6. Feed formulations based on energy and protein demands in white grouper (Epinephelus aeneus).. Brett. 2008. Helland. Aquaculture 237. 2779–2794. Bovaerts. 2007. Iwama. 1972. Fish Physiol..L.. 7.. The impact of varying feeding regimes on oxygen consumption and excretion of carbon dioxide and nitrogen in post-smolt Atlantic salmon Salmo salar L. P. Induction of ornithine–urea cycle enzymes and nitrogen metabolism and excretion in rainbow trout (Oncorhynchus mykiss) during early life stages. Nutr. Hatlen.. D. Aquaculture 139.. 16. Nutr. London. 2006. 1995... Fish.. Nutr.. J. G. T. Allan. 168–178. Glencross. Booth. D.... 2007. Anim. B. Evaluation of the influence of Lupinus angustifolius kernel meal on dietary nutrient and energy utilization efficiency by rainbow trout (Oncorhynchus mykiss).. Bureau. Energy and protein requirements for maintenance and growth in gilthead seabream (Sparus aurata L.. Physiological energetics. 1998.. X. Hatlen. A simple method for the measurement of daily feed intake of groups of fish in tanks. J. Sklan. Br. 1987. Evans. 99–114. Huisman. Effects of varying dietary protein and energy supply on growth. S. de Saeger. 2005. feed efficiency and digestibility in salmon (Salmo salar L. 37–53. D. but not protein or energy utilization efficiency of rainbow trout (Oncorhynchus mykiss). H. B. / Aquaculture 305 (2010) 150–158 Bodin... Hawkins. Assessment of lysine requirement for maximal protein accretion in Atlantic salmon using plant protein diets. Aquaculture 265.W. 1991.. Abboudi. Nutritional energetics of ruminants. Nutr. Pack.T. B. Mundheim. Bech-Andersen.. Edwards.. Helland et al. Bd. 194..D. G. 545–555. B.. Nutritional Energetics of Domestic Animals and Glossary of Energy Terms. Aquaculture 261. Maintenance requirement for digestible energy and efficiency of utilisation of digestible energy for retention in rainbow trout.. I.