Aquaculture Research, 2005, 36, 361^369

doi:10.1111/j.1365-2109.2004.01211.x

Enhancement of growth and feed utilization of the European sea bass (Dicentrarchus labrax) fed supplementary dietary salt in freshwater
an, Metin Kumlu, Mehmet K r & Gul Ayten Kiris Orhan Tufan Eroldog
Faculty of Fisheries, Department of Aquaculture, C ukurova University, Adana,Turkey
an, Faculty of Fisheries, Department of Aquaculture, C Correspondence: Dr O Tufan Eroldog ukurova University, 01330 Adana, Turkey. E-mail: mtufan@cu.edu.tr

Abstract
The eects of increased dietary sodium chloride supplement on growth, feed eciency (FE), feed dispensed rate and llet composition and morphological indices of European sea bass (initial weight of 4.9 g) were investigated in freshwater using ve dierent levels of supplemental salt (0 (control diet, with no salt supplemented), 1%, 3%, 5% and 9% of diet). Over the experimental period of 50 days, the nal weight of dietary salt (DS) groups was found to be approximately 19% higher than the control group. The nal weight, specic growth rate and FE were greatest at the moderate level (1^5%) of salt supplementation. Optimum level of DS inclusion was calculated as 3%. Feed eciency was signicantly enhanced with increasing salt level up to 5%. A negative relationship between daily feed intake and FE (r2 5 0.66) was observed. The average muscle ratio (MR) in groups fed 1^5% supplemented salt was around 51.4% whereas sh receiving control and 9% salty diets had 5.3% lower MR (48.7%) (Po0.05). However, no signicant eects of salt inclusion were found on viscera or hepatosomatic indices. Muscle ash and dry matter content slightly rose with increasing DS in the diet, but with no statistical dierence (P40.05). Similarly, no significant changes were observed in either muscle protein or lipid content among the groups (P40.05).

Keywords: dietary sodium chloride, Dicentrarchus labrax, freshwater, food intake, feed eciency

Introduction The European sea bass (Dicentrarchus labrax L.) is an economically important euryhaline marine sh that

frequently inhabits coastal waters of the Mediterranean Sea and is capable of tolerating both hypersa an line and freshwater (FW) environments (Eroldog an 2003; Saillant, Fostier, & Kumlu 2002; Eroldog Haray, Menu, Laureau,Thimonier & Chatain 2003). Juveniles of this species spend most of their rst 1976; year of life in estuaries or lagoons (Barnabe Kelley 1988). The rearing salinity commonly used in intensive sea bass culture systems ranges between oceanic salinity (35^40 ppt) as in the case of marine cage farming in the Mediterranean and low lagunar/ estuarine salinity commonly applied in inland systems. A number of studies examined the eects of salinity on growth performance and feed eciency (FE) of sea bass (Chervinski 1975; Alliot, Pastoureaud & an 1999, 2003). Moreover, in a Thebault 1983; Eroldog an, Kumlu and Aktas (2004) recent study, Eroldog compared food consumption rate in European sea bass cultured in both FW and seawater (SW) and concluded that those in FW consumed more food than sh in SW in order to maintain osmotic and ionic homeostasis during the passive water and ion movements between water and sh (Fontainhas-Fernandes, Monteiro, Gomes, Reis-Henriques & Coimbra 2000). In FW-adapted teleost, passive outward ux of ion from sh must be overcome by active uptake of ions (e.g. Na1, Cl , K1, Mg1 and Ca21) from the water and/or through the diet (Smith, Talbot & Eddy 1989; Karnaky 1998). It is reasonable to expect that diet is an important source of salts that could satisfy the osmoregulatory requirements of the sh in low-salinity water or even FW and might spare energy used for osmoregulation, thereby leaving more energy available for growth (Zaugg, Roley, Prentice, Gores

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 an et al. Aquaculture Research, 2005, 36, 361^369 Enhancement of growth and feed utilization of the D. labrax O T Eroldog

& Waknitz 1983; Gatlin, Macenzie, Craig & Neill 1992; Nandeesha, Gangadhar, Keshavanath & Varghese 2000; Fontainhas-Fernandes, RussellPinto, Gomes, Reis-Henriques & Coimbra 2001). Acclimation with salt-supplemented diets (at 4^8%) was found to be useful for non-smolting rainbow trout and other salmonids before release to the sea (Salman & Eddy 1987; Pelletier & Besner 1992; Staurnes & Finstad 2000), as such diets increase Na1^K1^ATPase activity and salinity tolerance in sh (Zaugg et al. 1983; Salman & Eddy 1988; Pelletier & Besner 1992). In addition to this fact, the results of some recently published reports on euryhaline species have indicated that growth and FE may be improved using salt-enriched diets (Gatlin et al. 1992; Nandeesha et al. 2000; Kim, Kim & Jeon 2001). However, inclusion of high levels of dietary salt (DS, 9^12%) in diets has negligible or even negative eects on growth and FE in salmonids (Salman & Eddy 1988; Pelletier & Besner 1992). It is reasonable to expect that DS could have a practical eect on the activation of the physiological mechanisms involved in salt secretion. Nandeesha and colleagues (2000) found that sh fed 1.5% NaCl showed best growth and this level of salt increased digestive enzyme activity of Cyprinus carpio and Cirrhinus mrigala juvenile. Furthermore, Smith, Eddy and Talbot (1995) observed that redirection of the Na1 to the tissue may reect enhanced uptake and distribution of nutrients (e.g. amino acids, glucose) for growth of salt-loaded sh after feeding. Thus, it may be presumed that sh fed supplementary salt and examination of the eect on the growth response curve can be positive. In view of the recent ndings obtained with the an 2003), DS apEuropean sea bass in FW (Eroldog pears to be imperative not only for continuance of osmotic and/or ionic homeostasis but also for the enhancement of growth and feed conversion eciency in this species. There is no information on the eect of DS supplementation on feed intake and growth in sea bass during the culture in FW. Our hypothesis is that a diet supplemented with salt oers a new approach to improving the growth of sea bass in FW where they need to balance their ion exchange for homeostatic mechanisms. Accordingly, this may be a presumable energy-saving strategy for growth as sh is virtually less stressed in terms of homeostasis in FW. The main objective of the present study was, therefore, to evaluate the eects of dierent levels of DS on growth and feed utilization in European sea bass during the culture in FW.

Materials and methods Experimental sh and design Five-month-old juvenile European sea bass (D. labrax) (4.9 0.5 g, 7.90 0.26 cm) cultured in 20 ppt in captivity were obtained from AKUVATUR Hatchery situated in the south of Turkey. After being reared in the same saline water for 1 week at 23 0.4 1C, all the sh were gradually acclimated to FW (0.4 g L 1) within 2 weeks by lowering the salinity with well water by approximately 1.5 g L 1 per day. Then, sh were acclimated to experimental conditions for another 2 weeks prior to the onset of the experiment. During this period, the sh were fed normal sea bass pellets twice daily during daylight. Two days before the end of this acclimation period, the sh were deprived of food to empty guts before the initial weight measurement. A total of 150 juveniles were divided into ve dierent DS groups (control diet, 1%, 3%, 5% and 9% of diet) to give triplicate replicates. Then, they were randomly stocked in15 circular polypropylene tanks (+ 57 cm; water volume, 70 L) tted with a continuous water ow-through system. During the culture period (50 days), the rearing water in each tank was enduringly saturated with oxygen by supplying air continuously through air-stones from an air-blower. Dissolved oxygen (9.3 1.2 mg L 1) was measured in the water from each tank, and never reduced below 8.2 mg L 1. Experimental diets and feeding trial In order to determine olfaction and taste of feed, one group of sea bass ngerlings (except of the experimental sh) was fed with test diets having enriched salt from 1% to 9%. This preliminary trial pointed out that an increasing salt level in the diet containing up to 9% salt was voluntarily acceptable to juvenile sea bass reared in FW. Before the onset of the study, the experimental sh were conditioned to a commercial sea bass feed for 2 weeks as explained above. Food was in the form of sinking extruded pellets, 3 mm in diameter, containing 48% protein, 12% fat, 12% dry matter and1.7% sodium chloride (NaCl), manufactured for sea bass (PINAR, Izmir, Turkey). The control group was fed with the feed described above. Test diets containing either 1%, 3%, 5% or 9% salt (by weight) were prepared by spraying NaCl onto the pellets while control diet having no additional NaCl but treated in the same way as other treatments. The amount of de-ionized water necessary for dissolving

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 an et al. Aquaculture Research, 2005, 36, 361^369 Enhancement of growth and feed utilization of the D. labrax O T Eroldog

the salt was slowly sprayed onto the diets. Then, water was allowed to evaporate from the feed under ventilated dried air condition at 22 1C. The salt (chlorine as NaCl) content of the experimental diets was analysed using the standard methods (Association Ocial Analytical Chemists (AOAC) 1995) and results showed that control, 1%, 3%, 5% and 9% NaClsupplemented diets contained an actual amount of 1.7 0.07%, 3.3 0.29%, 4.2 0.29%, 6.7 0.53% and 9.85 0.68% NaCl respectively. In order to nd NaCl leach loss of the diets, the triplicate groups of test diets were placed in the water for a period of 1, 3, 5, 10 and 15 s and then analysed using the methods explained above. All the experimental diets randomly assigned to ve groups of sh were kept in the vacuumed-box and held at 4 1C until required and thawed just before feeding to the sh. The sh were carefully fed to visual satiation, by hand, twice a day (rst 09:00^ 09:45 hours and then 17:00^17:45 hours). Every day, the remaining food was recovered and its weight subtracted from the initial amount. In order to follow good feeding practice and to avoid the possibility of inter-workererror, satiation feeding of experimental sh was carried out at the precise time by the same person throughout the culture period. During this feeding period, we found that sh consumed feed in 5^15 s, so that there was no leaching of salt from the test diets released.

Calculations Every10 days, all the sh from each tank were caught and weighed after blotting the sh on a paper towel. Before nal weight measurement, feeding was ceased for 24 h to empty the digestive tract, and then the sh were individually weighed to the nearest 0.05 mg. Growth performance, feed utilization parameters and all indices were described using the following equations: SGR 100 ln Wf ln Wi =days FDR 100 total feed dispensed= ln Wi ln Wf =2=days FE % wet weight gain=feed intake MR % 100 fillet weight=fish weight Survival % Nf Ni 100=Ni where SGR is the specic growth rate (% body weight/day), Wf and Wi are the nal and initial
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biomasses (g), t is time (duration of the respective growth period in days), FDR is the feed dispensed rate (% body weight/day), MR is the muscle ratio (%), FE is the feed eciency and for survival, Ni is the initial number of sh and Nf the nal number of sh. Feed utilization was examined by plotting FE of the tanks against their respective feed intakes. Before the beginning of the experiment,10 sh were randomly captured and killed to determine the initial llet composition of the pre-experimental sh. Random samples of ve sh from each replicate were caught and slaughtered by a sharp blow on the head at the end of the experiment. At the time of slaughtering, scales were removed mechanically and the whole llets (with skin and ribs) at both sides were manually lleted in a single cut along the backbone from the head towards the tail. The liver (gall bladder and bile were excluded from the liver weight) and visceral fat including the liver and gastrointestinal tract from the oesophagus to anus, were dissected free and weighed (to the nearest 0.01g) to estimate hepatosomatic (HSI) and viscerosomatic index (VSI). For proximate analysis, llets of ve sh from each tank were minced together and homogenized in a high-speed laboratory blender before the analysis. The proximate composition of homogenized sh llet was analysed in duplicate using standard laboratory methods essentially in accordance with Association Ocial Analytical Chemists (AOAC) (1984). Kjeldahls method was used to analyse percent llet protein (% of fresh llet weight) (Mattisek, Schnepel & Steiner 1988). Total lipid was estimated gravimetrically using the chloroform^methanol method of Bligh and Dyer (1959). Dry matter was determined by oven-drying at106 1C to constant weights; percent llet ash containing, e.g., non-combustible parts of bones was analysed from combusted llet samples in a furnace at 550 1C for 5 h.

Statistical analyses All statistical analyses were performed using J.M.Ps 3.2.1statistical software (Statistical Analysis Institute p (SAS) 1996). Arcsine transformations of percentage data were performed to achieve homogeneity of variance. The relationship between FDR, FE or DS level was analysed using a second-order polynomial t to the data: Y a bX cX 2 where Y is the FE or FDR, X is the supplemental DS (%) and a, b and c are constants determined by the

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 an et al. Aquaculture Research, 2005, 36, 361^369 Enhancement of growth and feed utilization of the D. labrax O T Eroldog

regression. The optimum salt level in the diet for FE or FDR (DSopt.FE or DSopt.FDR) was estimated as the zero solution [X 5 ^b(2c)^1] to the rst derivation of the polynomial equations. The FE and FDR at DSopt.FE and DSopt.FDR were dened as FEmax and FDRmin. Data were subjected to one-way analysis of variance (ANOVA), and data storage, data handling and graphics were carried out using Microsoft Excel (Microsoft, Redman, WA, USA). In all statistical tests, Po0.05 was taken as the level signicance, which was deter' s test after normality and mined using the Schee homogeneity (Bartletts test) of the data were performed (Sokal & Rholf 1981). Results Over the experimental period of 50 days, survival in all treatments was greater than 96%, with no signicant dierences among the groups fed dierent DS supplements (P40.05) (Table 1). No major dierences were found among the treatments in the nal coecient of variance (P40.05). Leaching test The results obtained from the leaching test show that salt leaching from the diets was 1.2% in 1s and increased up to 20.4% in 15 s post immersion. During the rst 5 s, the average leach loss was only 6.5% (Fig.1).

Growth performance of sea bass The present study showed that the growth performance of European sea bass ngerlings was signicantly enhanced when DS was used, regardless of the inclusion level (Table 1). Under satiation feeding conditions, growth performance results indicated that juveniles fed with a ratio of 1%, 3%, 5% and 9% supplemented salt had signicantly higher growth (nal body weight, SGR) than those fed with control diet (Table 1). The highest mean nal body weight of juveniles fed 3% DS was 1.3 times heavier than the control group. Specic growth rate of the sh fed 3% DS was highest, although there was no signicant dierence among the DS-loaded groups (P40.05). The wet weight values plotted against time suggest linear relationships (Fig. 2). Between the linear equation derived, there was no signicant (P40.05) dierence in the slopes of 1%, 3% and 5% salt-loaded groups. Over the period of 50 days, the highest growth rate as wet weight was detected at 1%, 3% and 5% supplemented-salt groups followed by those of control and 9% salt-supplemented group. To examine the relationship between SGR and supplemented- DS, the equation analysed with a parabolic regression were adopted as follows: SGR 1:254 0:5896DS 0:0864DS2 ; r2 0:87 1

Table 1 Final average body weight (FBW), specic growth rate (SGR), survival, coecient of variation (CV) of nal weight, daily feed intake (DFI), feed eciency (FE), feed dispensed rate (FDR) and some morphological traits of European sea bass ngerlings fed diet supplemented (by weight) with dietary salt (%) for 50 days
Dietary salt (%) Control
Growth performance FBW (g) SGR (% day1) CVnal DFI FE (%) FDR (BW day1) Survival (%) Morphological traits MRw HSIz VSI

11.8 1.7 19.3 142.8 48.8 1.7 100.0

0.9b 0.2b 4.9a 1.4a 0.9c 0.1b 0.0a

14.7 2.2 17.7 118.3 80.0 1.2 96.7

0.5a 0.1a 2.8a 2.9b 0.9ab 0.1a 5.8a

15.1 2.3 14.2 128.2 80.0 1.3 100.0

1.1a 0.1a 2.9a 5.9b 1.4ab 0.1a 0.0a

14.3 2.1 15.1 106.1 83.3 1.2 96.7

0.5a 0.1a 4.2a 3.8b 1.0a 0.2a 5.8a

14.1 2.1 14.9 147.8 62.1 1.6 96.7

0.7a 0.1a 6.2a 9.5a 1.2b 0.1b 5.8a

48.6 0.9b 1.3 0.3a 7.0 0.5a

51.2 0.7a 1.1 0.3a 6.5 0.8a

51.7 0.2a 1.0 0.2a 6.2 0.6a

51.3 0.9a 1.2 0.2a 6.8 0.3a

48.8 0.9b 1.3 0.4a 6.0 0.1a

Data are represented as means standard deviation of three replicates tanks. Within each column, means with no common letter are

signicantly dierent (Po0.05) (ANOVA, Tukey HSD). wMuscle ratio calculated as: MR 5 100 (llet weight/body weight). zHepatosomatic index calculated as: HSI 5 100 (liver weight, g)/(BW, g). Viscera somatic index calculated as: VSI 5 100 (viscera weight, g)/(BW, g).

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 an et al. Aquaculture Research, 2005, 36, 361^369 Enhancement of growth and feed utilization of the D. labrax O T Eroldog

25 y = 1.438x + 1.476 r = 0.89 Leach loss of NaCl (%) 20

Feed eciency and feed dispensed rate Feed eciency of juvenile sea bass was signicantly inuenced by salt in the diet. One-way ANOVA showed a signicant increase in FE of sh fed increasing levels of supplementary salt (1%, 3% and 5%) in the diet, in which FE was greatest at 5% inclusion level (83.3%) (Table 1). However, we could not detect any signicant dierence in FE between sh fed 1% and 3% salt supplements (P40.05). Fish fed the control diet had the lowest FE and were signicantly less ecient at converting feed to growth than were sh fed salted diet (Po0.05). Over the experimental period, FE of 1^5% salt-supplemented groups was 1.6^1.7 times higher than that of the control group. The FDR of the sh was inuenced by DS, with sh fed 1%, 3% and 5% salt inclusion having the lowest FDR (1.24, 1.28 and 1.27 respectively) (Po0.05), with the control (1.7) and 9% (1.6) salt-supplemented groups being higher than other treatments (Table 1). Taking into account the decrease in FDR with DS, the groups that received 1%, 3% and 5% ingested more feed than both the control and the 9% salt-received groups. In fact, throughout the experiment, the sh from those three groups ingested approximately 77.8 g of feed (weight gain FE), whereas sh fed control and 9% salty diet only consumed 33.9 and 57.1g respectively. The optimal DS for maximum FE and minimum FDR (DSopt.FE and DSopt.FDR) was calculated from the rst-order derivative of the parabolic regressions (i.e. when dFE/dDS or dFDR/dDS 5 0). for FE; FE 11:03 46:56DS 7:26DS2 2

15

10

0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Time (sec)

Figure 1 Leach loss of the salt (%) during dierent immersion times (each symbol represents a mean of three replicates).

18 16 14 Weight (g) 12 10 8 6 4 * 20 Days * 30 * 40 * 50 Control 1% 3% 5% 9% 3% 1% 5% 9% Control

10

r2 0:93 for FDR; FDR 2:31 0:73DS 0:12DS2

Figure 2 Growth in weight in juvenile European sea bass fed on dierent levels (control, 1%, 3%, 5% and 9%) of dietary salt (DS) over a period of 50 days. Dietary salt levels (% diet) are given on the right-hand side of the diagram. Asterisks on the X-axis denote signicant dierence between groups at Po0.05. Estimated equations for growth in weight of sea bass maintained in ve dierent DS (%): Y 5 b1 aX, where Y is the wet weight (g), X the time (days) and a and b are constants determined by the regression. The regression equations are: Y 5 2.47 1 1.45X, r2 5 0.97; (control). Y 5 1.94 1 1.98X, r2 5 0.96; (1% salt). Y 5 1.9712.16X, r2 5 0.98; (3% salt). Y 5 2.25 1 1.89X, r2 5 0.97; (5% salt). Y 5 2.65 1 1.79X, r2 5 0.98; (9% salt).

r2 0:89

In line with this equation, in order to obtain maximum growth, optimum supplementary DS was calculated as 3.4%.

In accordance with equations (2) and (3), optimum supplementary salt inclusion ratio DSopt.FE for maximum FE was calculated as 3.2%, while for minimum FDR this value was 3.0% (Fig. 3). As expected, the amount of feed ingested by the population increased as the sh grew. The average daily feed intakes (DFI: mean SD) during the experiment were 142.7, 118.3, 128.2, 109.8 and 147.8 g in the control, 1%, 3%, 5% and 9% supplemented salt in diets fed groups respectively. Feed eciency was negatively correlated with DFI (r2 5 0.66), which signies dierences in feed utilization (FE, gain:feed) (Fig. 4).

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90 85 80 Feed efficiency (%) 75 70 65 60 55 50 45 40 95 115 135 155 Daily feed intake (g) 1% 3% 5% 9% 175

y = -0.7133x + 163.02 r = 0.66

DS was only found in the MR (Table 1). Fish fed with ration of 1%, 3% and 5% supplemented salt were higher than control and 9% salt-loaded diets (Po0.05). The changes in esh composition of the sh at the start and end of the experiment are presented in Table 2. Analysis of llet nutrient composition of sea bass showed no signicant dierence among the treatments for moisture, lipid, protein and ash content during the growth period (P40.05).

Discussion The salt concentrations tested in the present study correspond roughly to typical DS levels found in salmon and tilapia diets commonly used (Tacon & DeSilva 1983; Pelletier & Besner 1992). The supplementation of high levels (4.5^11.6%) of NaCl to the diet has been reported to lower FE and inhibit growth in rainbow trout (Salman & Eddy1988). However, our results demonstrated that an increase in DS up to a level of 5% promotes both growth and feed utilization in sea bass in FW where they need to satisfy their ionic balance for osmoregulation. Excess supplementary salt inclusion (9%) has clearly decreased the FE compared with moderate salt levels (1% and 3%). The eects of DS on growth are controversial; studies with dierent sh species, i.e., Ictalurus punctatus (Murray & Andrews 1979), Salmon salar (Shaw, Saunders, Hall & Henderson 1979), and Oreochromis niloticus (Fontainhas-Fernandes et al. 2000), have shown negligible or even inverse eects of DS supplementation on growth. Moderate levels of salt (around 4% or 5%) have been reported to have benecial effects on growth and feed utilization by many authors (Orino & Kamizono 1975; Tacon & DeSilva 1983) for trout or carp and such supplementation has been demonstrated to be clearly useful for the European sea bass cultured in FW in our study. Growth performance of this species was signicantly enhanced by salt supplementation at low, medium or even high levels tested in our study (1^9%) in comparison with those received that the diet with no salt supplementation (control diet). Gatlin and colleagues (1992) stated that 2% salt supplementation in diet increased the growth rate of juvenile red drum, Sciaenops ocellatus, when reared at low salinity (FW and brackish water o6 ppt) for 8 weeks, but similar benets were not observed when such salt supplementation (2% and 10%) was tested in full-strength sea water (35 ppt). In our study, sh that received moderate levels of salt

Control

Figure 3 Relationship between daily feed intake and feed eciency for European sea bass fed dierent levels (control, 1%, 3%, 5% and 9%) of supplementary dietary salt. Each symbol signies an experimental group.

100 90 80 Feed efficiency (%) 70 60 50 40 30 20 10 0 Control 1 3 5 9 Supplementary dietary salt (%) c FE FDR a a a

4.0 3.5 3.0 b 2.5 2.0 1.5 1.0 0.5 0.0 FDR (body weight day )

Figure 4 Feed eciency and feed dispensed rate (FDR) of European sea bass fed dierent levels (control,1%, 3%,5% and 9%) of dietary salt. Each bar represents a mean SD (n 5 3 for each test dietary salt). Treatments marked with dierent superscripts are signicantly dierent from each other Po0.05.

Morphological traits and llet composition One-way ANOVA showed no signicant dierences among the treatments for either HSI orVSI. Although no eect of DS was observed on the HSI, there was a slight tendency for higher HSI in sh fed control and 9% DS-added diets as also reected in the feed intake data summarized in Table 1. Among morphological parameters, a signicant dierence of the eects of

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 an et al. Aquaculture Research, 2005, 36, 361^369 Enhancement of growth and feed utilization of the D. labrax O T Eroldog

Table 2 Mean initial and nal llet composition of European sea bass ngerlings fed diet-supplemented (by weight) dietary salt for 50 days
Dietary salt (%) Initial
Protein Dry matter Lipids Ash 20.8 26.7 3.1 2.5 0.1 0.1 0.2 0.4

Control
19.7 24.8 3.1 2.4 1.3 0.2 0.3 0.4

1
20.0 24.1 2.9 2.3 0.9 1.0 0.1 0.3

3
18.9 25.0 3.3 2.5 1.0 0.7 0.2 0.2

5
20.3 24.9 3.4 2.5 0.8 0.8 0.1 0.3

9
19.5 25.6 3.2 2.7 1.1 0.4 0.4 0.3

Data are represented as means standard deviation of three replicate tanks.

None of the treatments signicantly diered from each other (P40.05).

had greater growth rate than that those received higher levels (5% and 9%). In order to obtain maximum SGR, the sh needed to obtain 3.4% supplementary salt in their diet. Dietary salt at low saline water may increase the absorption of amino acids and/or satisfy other metabolic requirements by providing ions that the sh cannot suciently extract from hypotonic environments for whole-body Na1 and Cl homeostasis (Gatlin et al. 1992). In addition, an and colleagues (2004) indicated that sea Eroldog bass ngerlings reared in FW might spend more energy than those in SW for ionic and acid^base regulation. Apparently, the lost ions via the gill, kidney and guts in FW are regained by the sh through the DS included in their diets. Apart from the often-quoted metabolic cost of osmoregulation, growth dierences may be the result of changes in hormonal status, digestion, absorption and FE and metabolism. For instance, Smith and colleagues (1995) observed that redirection of Na1 to the tissues may reect enhanced uptake and distribution of nutrients for growth in FW rainbow trout, Oncorhynchus mykiss, after feeding with 12% salt-enriched diets. The regulation of feed intake is controlled using a satiation mechanism, while FE and digestibility was lineau, driven by enzymes in teleosts (Boujard, Ge ' s & Dutto 2000). The Corraze, Kaushik, Gasset, Cove present study highlights a link between the feed utilization and DS. Both FE and FDR were positively inuenced by the DS at 1^5% inclusion levels. Low FDR, high weight gain and FE mean that the sh fed dietary-supplemented salt (1^5%) have consumed less but have digested more food than the control and high level of DS (9%). The possible reason for low FE calculated for the group receiving 9% salt could be explained by protein dilution as suggested by Salman and Eddy (1988), Pelletier and Besner (1992) and also by energy expenditure for ionic regulation (Gatlin et al. 1992; Fontainhas-Fernandes et al. 2000). The

control diet had to be consumed more in order to extract higher level of ions to maintain osmotic an and homeostasis as also suggested by Eroldog colleagues (2004). Enhanced FE might have been the result of stimulated digestive enzymes by DS inclusion. Nandeesha and colleagues (2000) found an increase in protease, amylase and lipase activity and digestibility in C. carpio and C. mrigala fed salt-loaded diets. Similarly, Harpaz, Hakim, Slosman and Eroldo an (2004) found a partial increase in alkaline phosg phatase (AP) and lactase activity in pyloric caeca in Asian sea bass (Lates calcarifer) cultured in FW but not in salt water (20 ppt). Taking into account our results and those of the above researchers, we might also propose such a positive eect of DS inclusion on digestive enzyme activity in European sea bass. Leach loss of some substances such as minerals and vitamins from practical diets is true when introduced into the water environment and the loss may reach a substantial level by a longer immersion period (Slinger, Razzaque & Cho 1979). The preliminary leaching test of the diets showed that NaCl linearly leached out into the water when sprayed onto the pellets. This loss was 1.2% in 1s, reaching up to 20.4% in 15 s. Our pre-experimental observation demonstrated that the sh consumed the feed within the rst 5 s, during which the average NaCl leach loss was insignicant (only 6.5%). Muscle ratio is regarded as an important trait for the improvement of sh production eciency (Flick, Barua & Enriquez 1990). The average MR in the current study had improved substantially after using supplementary DS. The higher MR corresponded with the higher FE-presenting groups. We could not nd any signicant dierences among the groups in the HSI and VSI, which are used to assess the nutritional status of sh and are good predictors of the physiological condition of aquatic animals (Ng, Lu, an et al. 2004). Hashim & Ali 2000; Eroldog

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The levels of DS did not eect the proximate composition of the groups. Dry matter showed a slightly increasing trend with an increase in salt supplementation, which agrees with the observations of Tacon, Knox and Cowey (1984) and Nandeesha and colleagues (2000) in carp and of Orino and Kamizono (1975) in trout. Yet, Duston (1993) found that muscle water content in trout reared in FW was not signicantly aected by the experimental diets containing either control (0%) or 10% DS. Ash content marginally increased with supplemented salt, despite the fact that the results were not signicant. This agrees with the general pattern for trout (Orino & Kamizono 1975) and carp (Tacon et al. 1984) because of higher mineral content in the salty diet. Smith and colleagues (1995) also did not nd any dierence in muscle Na1 ion content of rainbow trout fed with diet containing salt levels between 0% and12%. The increasing protein and lipid content was generally noticed in rainbow trout (Orino & Kamizono1975) and common carp (Tacon et al.1984; Nandeesha et al. 2000), which is inconsistent with the ndings of lipid and protein content observed in the present study, in which these levels were more or less unchanged with increasing levels of DS. In conclusion, the present study has demonstrated that when cultured in FW, growth performance and FE of sea bass ngerlings could be signicantly enhanced by DS addition to practical diets. The optimum inclusion level of supplementary DS was calculated to be around 3%. Acknowledgments This research was supported by the Research Fund of the University of C ukurova, Turkey (FBE.2002.D.68). We thank Haluk Tuncer (Akuvatur AS) for supplying sea bass ngerlings used in the present study. References
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