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Aquaculture 265 (2007) 176 186

The effect of fish stocking density on the growth of California halibut

(Paralichthys californicus) juveniles
German E. Merino a,b,, Raul H. Piedrahita c , Douglas E. Conklin d
Departamento de Acuicultura, Facultad de Ciencias del Mar, Universidad Catolica del Norte, Larrondo 1281, Coquimbo, Chile
Centro de Estudios Avanzados en Zonas Aridas (CEAZA), Unidad Regional de Desarrollo Cientfico & Tecnolgico (CONICYT),
Colina El Pino s/n, La Serena, Chile
Biological and Agricultural Engineering Department, University of California, Davis, California, USA
Department of Animal Science, University of California, Davis, California, USA

Received 27 July 2006; received in revised form 12 January 2007; accepted 17 January 2007


The ability to raise California halibut (Paralichthys californicus) at a relatively high density, thus maximizing culture area
usage, is of particular importance for future commercial operations. The effect of stocking densities between 100 and 300% of
coverage area (PCA) on growth of juvenile (11.6 g) fish was tested in experiments that lasted 8 to 10 weeks. The experiments were
carried out in a recirculation system in small rectangular tanks and raceways with water depths under 6 cm. It was found that
maximum fish growth was achieved for fish stocked initially at 100% PCA. The results showed that California halibut can be
grown in shallow tanks and raceways at a relatively high stocking density without significantly compromising growth rate and
2007 Elsevier B.V. All rights reserved.

Keywords: Stocking density; California halibut; Paralichthys californicus; Surface area; Culture density; Flatfish

1. Introduction rainbow trout (Oncorhynchus mykiss) (Trzebioatowski

et al., 1981), Nile tilapia (Oreochromis niloticus) (Carro-
The ability to raise fish at a relatively high density, thus Anzalota and McGuinty, 1986), hamoor (Epinephelus
maximizing usage of the fish production infrastructure, is tauvina) (Abdullah et al., 1987), Arctic charr (Salvelinus
of importance to commercial aquaculture especially in alpinus) (Christiansen et al., 1992), African catfish
recirculation systems. Both positive and negative relation- (Clarias gariepinus) (Kaiser et al., 1995), gilthead sea
ships between stocking density and growth have been bream (Sparus aurata) (Canario et al., 1998), and lake
reported and the pattern of this interaction appears to be sturgeon (Acipenser fulvescens) (Fajfer et al., 1999).
species specific. The importance of stocking density on There are a few studies on the effect of stocking densities
fish growth has been reported for several species, such us on the growth of flatfish: turbot (Scophthalmus maximus)
(Martinez-Tapia and Fernandez-Pato, 1991; Irwin et al.,
Corresponding author. Departamento de Acuicultura, Facultad de
1999, 2003), olive flounder (Paralichthys olivaceus)
Ciencias del Mar, Universidad Catolica del Norte, Larrondo 1281, (Jeon et al., 1993), Atlantic halibut (Hyppoglossus
Coquimbo, Chile. Tel.: +56 51 209 767; fax: +56 51 209 764. hyppoglossus) (Bjrnsson, 1994), and summer flounder
E-mail address: (G.E. Merino). (Paralichthys dentatus) (King et al., 1998).
0044-8486/$ - see front matter 2007 Elsevier B.V. All rights reserved.
G.E. Merino et al. / Aquaculture 265 (2007) 176186 177

Flatfish species are bottom dwelling fish and to tance for future commercial operations. Consequently,
enhance growth and survival it has been recommended this research was planned to estimate the effect that
to provide an environment with a high bottom area to stocking density may have on the growth rate of
water volume ratio (Cripps and Poxton, 1992; Kaiser California halibut juveniles.
et al., 1995; iestad, 1999; Labatut and Olivares, 2004).
Stocking density for flatfish may be expressed either as 2. Materials and methods
biomass per unit of bottom area or as a percentage of
coverage area (PCA = percent ratio of total fish ventral A study was carried out to test the impact of three
area to total tank bottom area). Commercial land-based stocking densities nominally 100, 200, and 300%
facilities usually stock their fish at densities as high as PCA on fish performance. Stocking densities were
40 kg/m2 for olive flounder (Honda, 1998), 70 kg/m2 for tested in triplicate in small tanks also referred to as
sole (Liewes, 1984), and 120 kg/m2 for turbot (Person experimental tanks (Fig. 1), attached to the California
Le-Ruyet et al., 1991). Bjrnsson (1994) suggested that Halibut Recirculating Hatchery (Conklin et al., 2003;
the optimal stocking density for 2 kg Atlantic halibut is Merino, 2004). Fish morphometrics (total length, TL;
between 25 and 50 kg/m2 and between 50 and 100 kg/m2 total surface area with fins, TSA; body surface area
for 10 kg Atlantic halibut. However, the normal without fins, BSA) were determined at the beginning of
operational range for flatfish species varies from 15 to the experiment and at Week 8 by image analysis
50 kg/m2 depending on rearing conditions (Jones, 1981; (Merino, 2004). Fish average mass (W, g) was measured
Person Le-Ruyet et al., 1991; Liewes, 1984; Martinez- at the beginning of the experiment and every other week
Tapia and Fernandez-Pato, 1991; Mallekh et al., 1998; thereafter. The stocking density experiment lasted
Silva and Velez, 1998; Kikuchi, 2000). The suggested 10 weeks.
safe stocking density, as percentage of coverage area, Three of the raceways (Fig. 2) within the California
for flatfish is between 100 and 200% coverage area (Jeon Halibut Recirculating Hatchery (Conklin et al., 2003;
et al., 1993; Bjrnsson, 1994; King et al., 1998). Merino, 2004) were stocked at nominally 100, 200,
California halibut (Paralichthys californicus) is a 300% PCA as were the experimental tanks. The purpose
gregarious species in the wild and when stocked in was to validate the use of the experimental tanks by
tanks, therefore high stocking densities might be comparing W for the fish population from raceways and
possible for a commercial aquaculture system. To date, experimental tanks. Stocking densities were not repli-
no studies have been published that examine stocking cated for the raceways. After Week 6, the raceways
density for California halibut. The ability to raise stocked at 200 and 300% PCA started to have water
California halibut at a relatively high density, thus quality problems due to limitations in flow capacity and
maximizing culture area usage, is of particular impor- the comparison was terminated.

Fig. 1. Schematic diagram of an experimental tank used for the density experiment. All measurements are in cm. Each slot to introduce nets or weirs is
0.6 cm wide (measurement not shown). Figure is not to scale. Large arrow indicates the direction of the water flow. Water depth was 6.0 cm (not
shown). Tanks were built from dark grey acrylic plate, 0.6 cm thick.
178 G.E. Merino et al. / Aquaculture 265 (2007) 176186

Fig. 2. Schematic diagram of a raceway. Water flow was from left to right (arrow). Water depth was between 3.5 and 4.5 cm. Figure is not to scale.

2.1. Fish stock maintained at 6 cm by a weir located at the end of the

tank (second weir in Fig. 1). The first weir was placed
For this study 171 days post hatch (dph) weaned with the effluent screen (Fig. 1). A quiescent zone was
California halibut grown at the UC Davis California created between the weirs, allowing for observation of
Halibut Recirculating Hatchery were used. The juve- the amount of feed uneaten by the fish. Tanks were
niles (11.6 g wet mass) were reared in raceways on a flushed every day in the morning and before feeding by
recirculating seawater system with a constant tempera- removing the second weir for a few seconds. The
ture of 22 1 C and salinity of 30 g/L (Gadomski and function of the first weir was to keep a constant water
Caddell, 1991; Madon, 2002). The fish were fed daily at level of 2 cm during the flushing process.
1.2 % body mass with commercial dry pelleted feeds Water velocity in the experimental tanks stocked at
(EWOS Alpha #2; EWOS Pacific #3; http://www. 100% PCA was adjusted to 0.08 0.01 body lengths Light was provided by overhead fluorescent per second (bl/s) to match the corresponding velocity
tubes on a 16 L: 8 D (L = light; D = dark) photoperiod (0.09 bl/s) calculated for the 100% PCA raceway
(Boeuf and Le Bail, 1999; Klokseth and iestad, 1999). (Table 1). A velocity of 0.09 bl/s corresponded to a flow
rate of about 5 L/min for an experimental tank stocked at
2.2. Experimental tanks 100% PCA, resulting in a loading rate of 21.6 L/min kg.
This loading rate for the 100% PCA was used to
Rectangular tanks with a total internal length of determine the flow rates for the 200% (8 L/min) and
60 cm, and an effective experimental length of 41 cm, 300% (9 L/min) PCA experimental tanks, resulting in
were used for the stocking density experiment (Fig. 1). velocities of 0.16 0.01 bl/s and 0.25 0.01 bl/s for the
Three tank widths, 9.3 cm, 12.3 cm, and 15.3 cm, 200 and 300% PCA, respectively. Thereafter and as the
having a surface area of 381. 3 cm2, 504.3 cm2, and fish grew, the experimental tank flow rates were adjusted
627.3 cm2, respectively, were used to achieve the every other week to keep the initial relative swimming
desired stocking densities. Screens (5 mm aperture) velocities constant over time. Water velocities within
were placed at both ends of a tank to hold the fish within experimental tanks induced relative swimming velocities
the experimental area. The water depth in each tank was below 1 bl/s as recommended by Merino (2004).

Table 1
Culture conditions at time of stocking for raceways stocked at nominal densities of 100%, 200%, and 300% PCA
Nominal Biomass Fish W TSA TL Initial Flow Loading rate Velocity
PCA (%) (g) number (g) (cm2) (cm) PCA (%) (L/min) (L/min kg)
cm/s bl/s a
100 2404 214 11.23 30.6 9.9 107 9 3.7 1.1 0.09
200 4579 392 11.68 31.4 10.1 201 12 2.6 1.4 0.13
300 6795 616 11.03 30.0 9.9 306 15 2.2 1.8 0.16
Raceway surface area 6105 cm2.
Body length per second.
G.E. Merino et al. / Aquaculture 265 (2007) 176186 179

2.3. Raceways (Lifegard model AF2 from Aquatic Eco-Systems Inc; during the light hours
The raceways used were 2.41 m long, 0.28 m wide (0900 to 2200 h) 7 days/week for 10 weeks. Total fish
with a water depth between 3.5 and 4.5 cm (Fig. 2). A mass was measured at stocking time and then every
19 cm quiescent zone was created at the effluent section. other week to adjust the amount of feed, maintaining a
The tank surface area available for fish rearing, ration of 1.2% body mass per day. Fish mortalities were
excluding the quiescent zone was 6105 cm2. Cylin- replaced at sampling time with fish that matched the
drical nets (5 mm aperture) were placed at the influent average population morphological data for the respec-
and effluent to prevent fish from escaping and to provide tive replicate.
a frame support for a dissolved oxygen sensor.
Water flow rates in the raceways were set at stocking 2.5. Fish growth in raceways
time to have 5 mg/L of dissolved oxygen at the effluent,
resulting in 9, 12, and 15 L/min for the 100%, 200%, and This experiment was set up to compare growth
300% PCA treatment, respectively (Table 1). A maximum tendencies between fish stocked within the experimental
flow of 15 L/min per raceway could be achieved in the
system. The flow rates were maintained throughout the Table 2
six week duration of the test. Water velocities within the Summary of data from California halibut reared at three stocking
densities in experimental tanks for Week 0 (Initial), Week 8 (Final),
raceways induced relative swimming velocities below
and for selected parameters on Week 10 (Final at Week 10)
1 bl/s as recommended by Merino (2004).
Nominal stocking density as PCA (%)
2.4. Morphometrics and growth in experimental tanks 100 200 300
Average SD Average SD Average SD
California halibut juveniles to be stocked in the tanks W (g)
were obtained from a population (n = 1840) with an Initial 11.8 0.4 11.1 0.2 11.2 0.4
average W of 11.6 g, determined by counting and Final 23.9a 1.0 20.9b 0.4 19.0c 0.7
weighing the fish as a group. For simplicity and to Final at Week 10 28.5a 1.6 24.4b 0.6 21.2c 0.6
TL (cm)
minimize fish handling, the fish to be stocked in the
Initial 10.4 0.03 10.3 0.1 10.3 0.1
experimental tanks were selected from the population on Final 12.9a 0.1 12.5b 0.1 12.1c 0.2
the basis of total length (TL). The target TL for Wof 11.6 g TSA (cm2/fish)
was 10.0 cm and was determined as (Merino, 2004): Initial 33.3 0.3 33.0 0.6 32.9 0.4
Final 51.2a 1.2 48.1b 0.6 44.6c 1.2
TLcm 4:79W g0:302 ; r2 0:99: 1 BSA (cm2/fish)
Initial 21.7 0.5 21.5 0.4 21.4 0.4
The TSA corresponding to an average fish of 11.6 g Final 33.3a 0.2 31.9b 0.4 29.5c 0.8
CV (%)
was 31.2 cm2 and was determined as (Merino, 2004):
Initial 12.7a 2.3 12.9a 3.3 11.9a 1.8
Final 26.4a 1.8 23.0b 1.1 26.7a 1.1
TSAcm 7:08W g0:6056 ;
r 0:99:
2 Final at Week 10 29.4a 2.6 25.8a 3.6 30.1a 2.3
PCA (%)
The number of fish (N) to be stocked in each of the Initial 106 1 209 3 311 4
experimental tanks was estimated based on a TSA of Final 163 4 302 7 421 12
31.2 cm2/fish, the culture area available (A, cm2), and Final at Week 10 165 6 311 5 425 8
the target stocking density. The three nominal stocking Increment (%) 56 49 37
BFA (kg/m2)
densities, 100, 200 and 300% PCA, were established by Initial 3.8 0.1 7.1 0.1 10.6 0.4
stocking 20, 32, and 36 individuals, respectively, into Final 7.6 0.3 13.3 0.2 17.9 0.6
each of three replicate experimental tanks per treatment Final at Week 10 9.1 0.5 15.5 0.4 20.0 0.6
(Table 2). The percentage of covered area (PCA) was Increment (%) 140 118 89
determined as: BFV (kg/m3)
Initial 62.5 1.9 117.5 1.9 176.0 5.9
TSAcm2 N 100
Final 127.2 5.5 221.4 3.9 298.2 10.7
PCA% : 3 Final at Week 10 151.6 8.4 274.1 6.2 333.5 9.8
Acm2 Increment (%) 143 133 90
A different letter in a row indicates significant differences ( p b 0.05).
The fish were fed commercial dry pelleted feeds SD = Standard deviation; Increment = [((final at week 10) (initial)) /
(EWOS 1 and 2 mm) by automatic carrousel feeders (initial)] 100.
180 G.E. Merino et al. / Aquaculture 265 (2007) 176186

tanks and fish stocked within the raceways for 6 weeks. BSA) were gathered through image analysis (Merino,
California halibut juveniles were taken randomly from a 2004) only for fish within the experimental tanks at
population of 1580 fish with a mean W of 11.6 g and an initial stocking and at Week 8. Fish were not fed 24 h
estimated TSA of 31.2 cm2/fish (Eq. (2)). The number before sampling. Feeding was resumed the day after
of fish to be stocked in each raceway was determined sampling.
based on the calculated average TSA of 31.2 cm2/fish, Fish growth as percent gain with respect to the values
the tank culture area available (6105 cm2), and the at experiment initiation was calculated based on mass
nominal stocking density (100, 200, or 300% PCA). (PWG, %), total surface area (TSAG, cm2), and body
Each group of fish was weighed and the average W was surface area (BSAG, cm2). Other values determined
used to calculate the corresponding TSA (Eq. (2)) and were specific growth rate (SGR, %/day, Fonds et al.,
the actual PCA (Eq. (3)) (Table 1). 1995), feed efficiency (relative to feed offered, FE, g/g,
Raceways were cleaned daily in the morning before Forster and Ogata, 1996), coefficient of variation for the
feeding. Mortalities were removed and counted daily fish within the culture tanks (mass based, CV, %),
and were not replaced. Fish biomass was recorded every culture density as biomass of fish per area (BFA, kg/m2)
other week to adjust the amount of feed and maintain a or per water volume (BFV, kg/m3 ), and Fulton's
ration of 1.2% body mass per day. The fish were fed condition factor (K) (Innis, 1990) (Eq. (4)).
commercial dry pelleted feeds (EWOS 1 and 2 mm)
W g
by automatic belt feeders (model BFS12A from Aquatic K 100 4
Eco-Systems, Inc.) during the light hours (0900 to TLcm3
2200 h) 7 days/week for 6 weeks.
2.8. Statistical analysis
2.6. Water quality measurements
The response variables analyzed were W, TL, TSA,
Dissolved oxygen (DO) concentrations in the BSA, SGR, PWG, TSAG, BSAG, FE and CV. Response
effluents from the tanks were measured three times per variables were analyzed for significant differences
day. In the case of the raceways, effluent oxygen among treatments using a one-way complete model
concentrations were measured every 10 min with analysis of variance (Dean and Voss, 1999) with SAS
dissolved oxygen sensors (Sensorex model DO6000; statistical software, release 8.02 Level 02M0 (SAS Temperature was monitored Institute Inc., Cary, NC; The
with a copper-constantan thermocouple (Omega; stated null hypothesis (Ho) was that there is no placed in the system reservoir. difference in fish response between stocking density
Data from oxygen sensors and thermocouples were levels, and the corresponding alternative hypothesis (Hi)
recorded with a micrologger (Model 21X, Campbell was that at least one level is different. Significance was
Scientific, Inc.; TAN, analyzed using Duncan's multiple range test and
nitrite and nitrate were measured using a Hach Tukey's studentized range test at a significance of
Odyssey spectrophotometer (model DR/2500; http:// 0.05 (Dean and Voss, 1999). Readings of pH were taken with a The three raceways stocked at densities similar to
Fisher Scientific Accumet pHmeter (model 50; https:// those of the experimental tanks were used to validate the Alkalinity was measured by use of the small tanks by comparing the tendencies for
titration (La Motte Chemical test kit, model WAT-DR; growth data from raceways and experimental tanks Salinity was measured with a stocked at similar densities.
YSI SCT meter (model 33; TAN,
nitrite, nitrate, pH, salinity, and alkalinity were moni- 3. Results
tored daily at 9:00 h before fish feeding. Sodium
bicarbonate was added daily at 9:00 h and 17:00 h to Salinity during the experiments was 30.1 1.3 g/L,
maintain alkalinity. and temperature was 22.4 0.7 C. Temperature in-
creased during Week 6 and remained at 24.1 1.0 C
2.7. Fish sampling until Week 9 due to a chiller failure. During Weeks 9 and
10 the temperature was lowered to 19.6 1.2 C. During
All fish were weighed as groups every other week to the higher temperature period fish behaved normally but
determine biomass so that feeding rations could be TAN concentration increased from less than 1.0 mg/L to
adjusted. Individual morphological data (TL; TSA; and 5.5 mg/L. Nitrite nitrogen concentration varied between
G.E. Merino et al. / Aquaculture 265 (2007) 176186 181

Table 2 lists the growth performance of the fish reared

at the three stocking densities. At stocking time the fish
were chosen to provide a uniform TSA. There were no
significant differences ( p N 0.05) between treatment
means for W, TL, TSA, BSA, and CV at the start of the
experiment. Mean W was significantly different for the
three treatments for Weeks 8 ( p b 0.05) and 10 ( p b 0.05),
with the mean W for the 100% PCA treatment being
greater than that for the 200% PCA treatment, which was
in turn greater than that for the 300% PCA (Table 2).
Significant differences ( p b 0.05) between treatments
were seen on Week 8 for TL, TSA, BSA, and CV
(Table 2). The coefficient of variation (CV) increased
significantly ( p b 0.05) from the start of the experiment
to Week 10 for the three stocking densities. However,
there was no clear trend in terms of the evolution over
time of CV differences between the three stocking
densities. In fact, although CVs were significantly
Fig. 3. Mean mass (W ) of California halibut juveniles reared at three different ( p b 0.05) among stocking densities at Week
densities during 10 weeks. Vertical lines represent standard deviations. 8, no differences were found at Week 10 (Table 2).
At the end of the experiment the largest fish were
those stocked originally at 100% PCA, and the smallest
0.2 and 0.9 mg/L. Water pH remained unchanged over those stocked at 300% PCA. The increase in fish
time, having an average of 7.3 0.2. Alkalinity varied biomass per area (BFA) at Week 10 was 140%, 118%,
between 2 and 3 meq/L and had an average value of and 89% for the 100% PCA, 200% PCA, and 300%
2.46 0.3 meq/L. PCA, respectively (Table 2). The increase in fish
The lowest dissolved oxygen in the influent water to biomass per volume (BFV) at Week 10 was 143%,
both the experimental tanks and raceways at stocking time 133%, and 90% for the 100% PCA, 200% PCA, and
was 6.4 0.1 mg/L. Effluent dissolved oxygen concentra- 300% PCA, respectively (Table 2). Growth in W for the
tions from the raceways reached 4.2 0.3, 4.4 0.4, and duration of the experiment was represented well by
4.9 0.4 mg O2/L for 300% PCA, 200% PCA, and 100% exponential curves (Fig. 3).
PCA, respectively, during Week 6, when the raceway Average PWG, SGR, TSAG, and BSAG were
experiment was terminated. The lowest dissolved oxygen affected significantly by stocking density (Table 3).
concentration in the effluents from the experimental tanks Significant decreases ( p b 0.05) in PWG, SGR, TSAG,
was 5.8 0.2 mg/L and was observed at the end of the and BSAG were observed as density was increased.
experiment (Week 10) in the tanks stocked at 300% PCA.

3.1. Morphometrics and growth in experimental tanks

Table 3
Fish growth and feed efficiency for California halibut reared at three
A total of five fish were replaced in replicates of the stocking densities for 8 weeks in experimental tanks (mean valuestandard
300% PCA treatment, two of which died in the tanks deviation)
and the other three jumped out of the tanks. Five fish Nominal stocking density (%)
were replaced in replicates of the 200% PCA treatment, 100 200 300
one of which died in the tank and the others jumped out
Average SD Average SD Average SD
of the culture tanks. No fish had to be replaced in
replicates of the 100% PCA treatment. PWG (%) 103.5a 3.8 88.4b 3.4 69.5c 6.4
SGR (%/d) 1.18a 0.03 1.06b 0.03 0.88c 0.06
Some fish in the 300% PCA treatment tended to
TSAG (%) 53.9a 3.9 45.9b 1.4 35.5c 2.7
attach themselves to the side walls of the tanks. A total BSAG (%) 53.4a 2.3 48.2a 1.6 37.7b 2.8
of three fish with damaged tails were found in the 300% FE (g biomass/g feed) 1.22a 0.03 1.10b 0.02 0.89c 0.06
PCA treatment tanks from Weeks 2 to 10. No injured Values followed by a different letter on the same row are significantly
fish were seen in any of the 100% and 200% PCA different ( p b 0.05).
treatment tanks. SD = Standard deviation.
182 G.E. Merino et al. / Aquaculture 265 (2007) 176186

The FE was affected significantly by stocking similar and larger than for the fish stocked at 300%
density (Table 3). A significant decrease ( p b 0.05) in PCA.
FE occurred as stocking density was increased, from a
maximum of 1.22 g/g in fish initially stocked at 100% 4. Discussion
PCA to 0.89 g/g for fish stocked at 300% PCA (Table 3).
The condition factor, K, at stocking time for all PCAs Water velocities within the tanks and raceways
was 1.0. At Week 8, K was 1.11, 1.06 and 1.08 for 100, induced swimming velocities which were less than
200, and 300% PCA, respectively. 1.0 bl/s. Merino (2004) found that California halibut
juveniles grew faster and made more efficient use of the
3.2. Fish growth comparison between raceways and feed provided at relative swimming velocities of 0.5 and
experimental tanks 1.0 bl/s than at 1.5 bl/s. Bengston and Alves (2002)
found that summer flounder (124 g, 257 g, and 387 g),
This experiment was terminated on Week 6 due to grew best when reared at 0.5 bl/s in comparison to
low oxygen concentrations in the raceways stocked at those grown at relative swimming velocities less than
200 and 300% PCA. Growth data for fish reared at three 0.3 bl/s or greater than 1.3 bl/s. Ogata and Oku (2000)
stocking densities in the raceways and experimental found that water flow velocities corresponding to a
tanks are shown in Table 4. relative swimming velocity beyond 2.1 bl/s (19.5 to
Fish were stocked at about the same BFA in both 23.5 C) led to growth retardation and lower feed
vessel types but water depths were lower in the efficiency for all olive flounder sizes tested and reported
raceways resulting in higher volumetric densities in that the optimal water flow velocity for growth occurred
comparison to the experimental tanks (Table 4). The at about 1.0 bl/s for a 12.5 cm/body length (5.7 g/fish).
overall tendency for W at Week 6 was to decrease with Therefore, there is enough evidence to support that the
stocking density with the exception of fish stocked in swimming velocities used in these experiments do not
raceways at 100 and 200% PCA for which the W were affect the growth performance of California halibut
Table 4 All water quality parameters were within values
Growth summary and observed volumetric density for California recommended in the literature for other flatfish species.
halibut juveniles reared during six weeks at three stocking densities
In the case of TAN, the worst condition measured was
within two types of culture vessels
5.5 mg TAN/L (salinity 30.1 g/L; 24.1 C; pH 7.3)
Nominal stocking density (%) which is equivalent to about 0.05 mg/L un-ionized
100 200 300 ammonia as nitrogen (NH3N), Juvenile turbot ( 20 g)
Average SD Average SD Average SD reduced their food intake when un-ionized ammonia
Raceways was over 0.117 mg NH3N/L, and a reduction in body
W (g) mass gain per day occurred when the level was over
Initial 11.2 11.7 11.0 0.108 mg NH3N/L (pH 8, 16 C, 28 g/L salinity)
Final 20.6 20.5 18.2
(Rasmussen and Korsgaard, 1996). Person-Le Ruyet
BFA (kg/m2)
et al. (1997) reported that for 13, 23, and 104 g turbot
Initial 3.9 7.5 11.1
Final 7.1 13.1 17.5 reared in water having in the effluent over 80% oxygen
BFV (kg/m3) saturation (6 mg/L), the growth was not affected at
Initial 101 165 228 concentrations of 0.21, 0.18, and 0.09 mg NH3N/L,
Final 181 286 359
while growth stopped immediately for all groups above
0.8 mg NH3N/L ( pH 8, 17 C, 34.5 g/L salinity).
Experimental tanks
W (g) Turbot survival was not affected at levels below 0.4 mg
Initial 11.8 0.4 11.1 0.2 11.2 0.4 NH3N/L (13 mg TAN/L) (Person-Le Ruyet et al.,
Final 20.7 0.9 18.3 0.4 17.0 0.7 1997). Therefore the worst concentration of un-ionized
BFA (kg/m2) ammonia as nitrogen recorded in this experiment is
Initial 3.7 0.1 7.1 0.1 10.6 0.4
within a safe range. In the case of dissolved oxygen, the
Final 5.6 0.2 9.8 0.4 14.2 0.4
BFV (kg/m3) stocking density trial carried out in the raceways was
Initial 62.5 1.9 117.5 1.9 176.0 5.9 terminated at the end of Week 6 when dissolved oxygen
Final 109.8 4.7 193.9 4.5 267.6 10.3 in the raceway effluent dropped below 60% of oxygen
Tank depth was 6 cm and raceway depth was 3.64.5 cm. saturation, the value recommended as an allowable limit
There was no SD for raceways as they were not replicated. for turbot (Brown et al., 1984). Effluent dissolved
G.E. Merino et al. / Aquaculture 265 (2007) 176186 183

oxygen in the experimental tanks remained above 75% 298.2 kg/m3) (Table 2). Fish held at the lowest densities
of saturation throughout the experiment. had the largest SGR (Table 3). Most studies on flatfish
Fish stocked in the experimental tanks at 200% and stocking densities have found that higher densities result
300% PCA did not eat all the feed offered between in lower growth. Irwin et al. (1999) reported that turbot,
Weeks 8 and 10. The increase in pellets left in the with initial biomass of 8.62 g/fish, were stocked at four
quiescent zones of the tanks loaded at 200% and 300% different densities (10%, 14%, 17%, and 21% PCA),
PCA started when the stocking density reached 283% and after 45 days fish reared at the higher stocking
PCA and 397% PCA, respectively. California halibut densities (final density 149.3% PCA) showed signifi-
reared in the experimental tanks had a drop in SGR cantly slower growth rates than those held at lower
during the last 2 weeks of experimentation, regardless of densities (final density 67.4% PCA). Jeon et al. (1993)
the tank stocking density. A reduction of feeding activity reported that olive flounder with initial biomass of 42 g/
and a drop in SGR can indicate a possible stressful fish were stocked at five different densities (33%, 50%,
condition. 100%, 200%, and 300% PCA), and after 35 days of
The culture density of flatfish is frequently reported experimentation it was found that fish at the highest
as the PCA (Bjrnsson, 1994; Klokseth and iestad, stocking density (final density 460% PCA) showed a
1999). A common practice to estimate surface area of significantly slower growth rate than those held at the
flatfish in aquaculture is by placing the fish over a paper lowest density (final density 45% PCA). King et al.
grid, drawing the fish edges, and counting the area (1998) summer flounder with initial biomass of 7.8 g/
enclosed by the drawing (Bjrnsson, 1994; King et al., fish, were stocked at three different densities (100%,
1998; Fairchild and Howell, 2001). The limits of this 150%, and 200% PCA), and after 58 days the fish reared
current methodology are numerous. Foremost is that at the highest stocking density (final density 493%
since the process is labor intensive and time consuming PCA) showed significantly slower growth rates than
only a few animals can be sampled. Sampled fish are those held at the lowest density (final density 355%
subjected to excessive manipulation, with a high PCA).
potential for injury. Merino (2004) used photographic Feed efficiency was affected when California halibut
and computer image analysis techniques to perform an were stocked at higher densities as has been described
accurate and fast method to estimate surface area of for olive flounder (Jeon et al., 1993). In the current
flatfish which he called total surface area (TSA) (area of study, a considerable amount of feed was found in the
fish edges) as well as body surface area (BSA) quiescent zone of tanks stocked at 200% and 300% PCA
(excluding the fins) of California halibut. starting on Week 8. There are reports that with rising
Fulton's condition factor (K ) is an indicator of stocking density, an increase in aggressive behavior,
general fish well-being. California halibut in the present growth, and feed efficiency was observed for the
study had a K value between 1.06 and 1.11, which dominant fish while stress, fin damage, and metabolic
according to Fulton's condition definition will be rates increased in subordinate fish (Refstie and Kittel-
considered as a healthy fish. sen, 1976; Purser and Hart, 1991; Canario et al., 1998).
A study done with turbot did not find differences on
4.1. Morphometrics and growth in experimental tanks growth nor aggressive behavior between equally sized
or mixed animals (Sunde et al., 1998). Fish stocked in
Morphometric analysis was planned originally for a the present study were pre-selected by size and only one
period of 10 weeks, but a mechanical problem with the case of damaged tail was observed in each of the 300%
chiller resulted in an increase in the water temperature PCA replicates. Bjrnsson (1994) found that for
from 22 to 24 C during Week 6. Therefore, it was Atlantic halibut, more of the feed consumed was used
decided to do a morphometric analysis at the next for metabolism than for growth with increasing stocking
scheduled sampling (beginning of Week 8). Madon densities. Thus it is possible that an increase in
(2002) who experimented with California halibut metabolic rate, which was not measured, at higher
juveniles of similar sizes to the ones used in this stocking densities might have been the cause for the feed
research did not detect differences in growth for efficiency and growth reduction found here with
temperatures ranging from 20 to 28 C. California halibut.
In the experiments described here, fish stocked at A direct effect of stocking density on growth
100% PCA (final 163% PCA; 127.2 kg/m3) grew larger variability has been reported by a number of authors
than those stocked at 200% PCA (final 302% PCA; (Andrews et al., 1971; Irwin et al., 1999; Lambert and
221.4 kg/m3 ) and 300% PCA (final 421% PCA; Dutil, 2001). The relative size difference between
184 G.E. Merino et al. / Aquaculture 265 (2007) 176186

members of a population usually increases with angular tanks and raceways at a relatively high stocking
increasing stocking density due to social interactions, density without significantly compromising survivabil-
development of hierarchies, and establishment of ity. In this study, the number of fish was held constant
territorial borders (Lambert and Dutil, 2001). Further- for the duration of the experiment, and biomass density
more, an increase in the CV for fish within a population (BFA and BFV) was allowed to increase with time due
is considered to be indicative of the establishment of to the growth of the fish. Initial stocking densities of
hierarchies (Jobling, 1994; Irwin et al., 1999; Lambert 100% PCA produced a higher individual fish growth
and Dutil, 2001). The CV for turbot was found to over time than the other stocking densities tested, which
increase more rapidly in groups held at higher stocking indicates that stocking density does affect the growth of
densities (Irwin et al., 1999). It was also found that the California halibut juveniles. The results indicate that, of
CV for turbot increased with increasing size of fish both the densities tested, the optimal one is somewhere
for graded and ungraded groups (Sunde et al., 1998). In between 100% and 200% PCA. Therefore, tanks with
this study, the CV increased significantly for all groups California halibut juveniles should be split when the
of California halibut during the 10 weeks of experi- stocking density reaches a maximum target PCA to
mentation, indicating that growth rates were affected at reduce the BFA and prevent a significant reduction in
all stocking densities analyzed. However, there was no SGR.
clear trend in terms of the evolution over time of CV Volumetric biomass densities for California halibut
differences among stocking densities (Table 2). juveniles can be relatively high. In this study, BFV
reached values between 151.6 and 333.5 kg/m3, which are
4.2. Fish growth comparison between raceways and larger than those reported for other flatfish (Irwin et al.,
experimental tanks 1999; Iglesias et al., 1987; Jones et al., 1981; Labatut and
Olivares, 2004). The high BFV reached in this study were
Experimental tanks were used in triplicate to set up a the result of rearing the fish in shallow tanks.
density experiment and three raceways were stocked at In this study relatively low BFA of 9.1 and 20.0 kg/m2
the same densities (100%, 200%, and 300% PCA) and were tested due to the small size of the fish used, but
used as controls against the experimental tanks. Results work with olive flounder (Honda et al., 1993; Honda,
for the experimental tanks agreed well with those from 1998), turbot (Person Le-Ruyet et al., 1991), and
the raceways (Table 4). Raceways, which were 5.6 times Atlantic halibut (Bjrnsson, 1994) suggest that values
larger, 23 times wider, and stocked with 9 to 16 times of 48 (for a 120 g fish), 25 to 50 kg/m2 (for fish N 300 g),
more biomass than experimental tanks had growth and 25 to 50 (for a 2 kg fish), respectively, are possible.
results similar to those in the tanks. The BFA can be estimated by knowing W, the
Because of the shallowness of the water in the tanks corresponding TSA, and the PCA. The TSA can be
and raceways used, BFV in this research (Table 4) are measured for flatfish by using image analysis (Merino,
larger than the 75 kg/m3 maximum reported for turbot 2004). For California halibut, TSA can be estimated
(Irwin et al., 1999; Martinez-Tapia and Fernandez-Pato, using Eq. (2) (Merino, 2004). Having W and TSA, an
1991). In addition BFV and BFA in the current research
were also larger than the 200.8 kg/m3 and 8.03 kg/m2,
respectively, reported recently by Labatut and Olivares
(2004) for 7.63 g turbot reared at 262 PCA% in shallow
raceways with a rearing area of 36,000 cm2 and a
velocity of 0.64 bl/s. Areal densities reported for conven-
tional aquaculture vessels are up to 29.5 kg/m2 for
juvenile summer flounder (King et al., 1998), 36.3 kg/m2
for olive flounder (Chang et al., 1995), 68 kg/m2 for turbot
(Martinez-Tapia and Fernandez-Pato, 1991), and 95 kg/
m2 for halibut (Bjrnsson, 1994) which suggests that
flatfish can be raised at high densities.

4.3. Practical implications

The practical implications of the present results are Fig. 4. Projected stocking density of California halibut corresponding
that California halibut can be grown in shallow rect- to 100%, 200%, and 300% PCA related to biomass of fish.
G.E. Merino et al. / Aquaculture 265 (2007) 176186 185

equation can be written that relates fish biomass (W ) State Resources Agency. The views expressed herein are
with stocking density: those of the authors and do not necessarily reflect the
  views of NOAA or any of its sub-agencies. The U.S.
W g Government is authorized to reproduce and distribute
BFA a k 5
TSAcm2 for governmental purposes.
Other sponsors were the Chilean Government
where is the stocking factor and is equal 1 for 100% through the Beca Presidente de la Republica, the
PCA, 2 for 200% PCA, and so on; and k is a conversion Organization of American States (OAS) through a
factor (10) to express g/cm2 as kg/m2. Increases in W will fellowship under the Regular Training Program (PRA),
result in the corresponding rises in BFA for a given PCA and the University of California at Davis through a
as shown in Fig. 4. The graph does not represent growth, Jastro-Shields Research Scholarship.
merely the calculated biomass of fish stocked per surface
area (BFA) for a given fish size (W ) and desired PCA. For References
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