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 Aquaculture 264 (2007) 418 – 427
www.elsevier.com/locate/aqua-online

Swimming activity and feeding behaviour of larval European sea

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bass (Dicentrarchus labrax L): Effects of ontogeny and
increasing food density

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V. Georgalas, S. Malavasi ⁎, P. Franzoi, P. Torricelli
Dipartimento di Scienze Ambientali, Università Ca' Foscari di Venezia, Castello 2737/b, Campo della Celestia, 30122 Venezia, Italy
Received 9 December 2005; received in revised form 26 April 2006; accepted 27 September 2006

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Abstract
on
The behaviour of European sea bass larvae reared with the mesocosm technique was analysed in relation to ontogeny and increasing
food density, with a particular focus on swimming and feeding activities. The behaviour of groups of larvae (20 animals each) belonging
to three age classes (10, 20 and 30 days post-hatching) was video-recorded in experimental tanks under controlled laboratory conditions.
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In each replicate, three different food densities, in terms of number of naupliar Artemia/l, were obtained. The videotapes analysis allowed
the identification of 6 behavioural units (larval MAPs) and their quantification in terms of either frequency (number/min), or duration
(percent of time spent in that activity with respect to the total time of observation). Behavioural variation was analysed in relation both to
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age and food density. Results revealed that swimming activity increased significantly with age, especially between the 10 and the 20 days
post-hatching, whereas resting activities decreased with age and, in some cases, also with food density. Frequency of feeding behaviours
decreased with age if they were analysed singly, but when the association between aiming posture (Sigmoid-posture) and attacks was
considered, results showed that these behaviours increased with both age and food density. Further, the increase of feeding efficiency
with age was also confirmed by the increase in the number of Artemia nauplii/individual across the three age groups. Results are
discussed in the light of both a comparative analysis conducted with the available data on the larval behaviour of other marine fish
species, and their potential use for the assessment of the quality of the reared fish, also in relation to the rearing techniques.
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© 2006 Elsevier B.V. All rights reserved.

Keywords: Sea bass larvae; Feeding behaviour; Swimming activity; Ontogeny

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1. Introduction aquaculture and fisheries (Ardizzone et al., 1988). Within

this context, a strong effort is devoted to seed production
Along the North Adriatic coast and especially in in order to restock the natural populations (Cataudella
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brackish lagoons, the juvenile stages of the European sea
bass Dicentrarchus labrax (L.) are a valuable biological
resource, being the object of specialised forms of
⁎ Corresponding author. Tel.: +39 0412347739; fax: +39
0415281494.
E-mail address: mala@unive.it (S. Malavasi).
0044-8486/$ - see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2006.09.035
et al., 2001). Juveniles are, in some cases, produced by
means of the mesocosm technique, based on large
volumes, low larval density and hydrodynamic conditions
more similar to that of the natural environment (Catau-
della et al., 2001; Roncarati et al., 2001). It is documented
that this technique allows the production of fish similar, in
terms of general morphology, to the wild counterparts
(Koumoundouros et al., 1997; Cataudella et al., 2001).
 V. Georgalas et al. / Aquaculture 264 (2007) 418–427
These results suggest that the conditions affecting the
larval phase play a fundamental role for the proper
development of fish. Although ontogeny of some
morphological structures and the relationships between
rearing conditions and body shape in this species have
been investigated in previous studies (Loy et al., 2000;
Diaz et al., 2003), behavioural studies of the larval stage
of this species are lacking.
As stated by Brown et al. (1997), behavioural
analyses could promote the design of better rearing
environments for larval production. This kind of
behavioural data could be combined with morphological
and genetic indicators to provide more complete tools
for the assessment of the quality of the produced fish
and for the improvement of rearing systems.
The aim of this study is to provide a quantitative
analysis of the behaviour of the larval stage of the
European sea bass reared with the mesocosm technique,
with a particular focus on feeding and swimming
activities. The majority of fish larvae are selective,
on
visual predatory planctivorous, whose performance is
affected by several factors, such as ontogeny, turbulence,
light and food density (Puvanedran and Brown, 2002;
Utne-Palm and Stiansen, 2002). The ability to modulate
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the feeding behaviour in relation to prey availability has
been considered an important adaptation to a changing
environment (Munk, 1995). In the present study, feeding
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and swimming behaviours of sea bass larvae were
recorded and quantified under controlled experimental
conditions, and their variation was analysed in relation to
ontogeny (age, in terms of post-hatching days) and food
availability (increasing density of naupliar Artemia). In
particular, the variation of behaviour as a response to a
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short term increase of prey availability was tested in
larvae of three different age groups. Results were
discussed in the light both of their potential use for the
assessment of the quality of the reared fish, and of a
o
comparative analysis conducted with the available data
on the larval behaviour of other marine fish species.
th
2. Material and methods
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2.1. Experimental fish and housing conditions
Larvae of sea bass were obtained from induced
spawning of captive fish in the hatchery facilities at Val
Figheri (Southern Venice Lagoon, northern Italy, 45° 18′
N; 12° 10′ E). The broodstock had spent 7–8 years in
captivity and originated from the Venice Lagoon. This
hatchery facility uses a semi-extensive larval rearing
technique (Cataudella et al., 2001; Shields, 2001). The sea
bass larvae in this case were held in 64 m3 circular
419
polyester tanks situated indoors at a larval density of 5–
6 ind/l. These tanks were filled to a depth of 1.5–1.7 m
and were artificially illuminated at a photoperiod of
24 D:0 N. Light intensity on water level at the center of the
rearing tanks measured with a digital luxmeter (MITEK,
MK5334) 1100–1300 lx. Endogenous production of
microalgae and zooplankton was sustained for the entire
duration of larval rearing and Artemia nauplii was added
py
in order to maintain a stable concentration of Artemia
nauplii/l (see below for details and concentrations for
every given age). No water exchange in the rearing tanks
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took place during the first 35 days of rearing.
Larvae from one batch, stocked into a single
mesocosm tank, were sampled on 10, 20 and 30 post-
hatching for video analysis of behaviour.
2.2. Experimental procedure
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The experimental tanks were small glass-tanks
(3.37 l; 15 × 15 × 15 cm), filled with filtered salt water
with the same characteristics of that of the rearing tanks
(salinity 20). Sheets of thick, dark-blue coloured paper,
covered the rear and lateral walls of each experimental
tank to minimise visual disturbance and provide an
adequate visual contrast with the fish silhouette to
facilitate videotapes analysis. One 60 W lamp was
suspended 40 cm over the tank to produce about 1200/
1300 lx light intensity (measured at the water surface
with a digital luxmeter (MITEK, MK5334). Light
intensity was measured before each experimental
replicate and was compared among experimental
groups, being 1215 lx (± 150) in the 10 days old larvae,
1279 lx (± 205) in the 20 days old larvae and 1301 lx
(± 214) in the 30 days old larvae, and there was no
statistically significant difference among the three
groups (Kruskal–Wallis, n = 28, P N 0.05). Water tem-
perature was also checked at the end of each replicate by
means of a digital thermometer (Hanna Woonsocket,
RI02895), and also in this case no difference among
experimental groups was detected (mean temperatures
19.1 ± 0.22 °C; Kruskal–Wallis, n = 28, P N 0.05).
The adopted experimental design aimed to test for the
effects of ontogeny and increasing prey availability on the
larval behaviour. Variation of larval behaviour was
analysed across three age groups treatments (10, 20 and
30 days post-hatching), each consisting of 8 (10 days old
larvae) or 10 (20 and 30 days old larvae) groups of 20
larvae, for a total of 28 replicates. Within each group of 20
larvae, the behavioural response towards increasing prey
availability was tested by increasing progressively prey
density from 1/4 of the prey density present in the rearing
tank (d0), to a density equal to half the prey density
420 V. Georgalas et al. / Aquaculture 264 (2007) 418–427
present in the rearing tank (d1) and, finally, to a density
equal to the actual prey density present in the rearing tank
(d2). Therefore, the assessment of the behavioural
ontogeny was performed by comparing larval MAPs
across the three independent age groups, whereas the
behavioural variation due to increased prey availability
was analysed by testing for the temporal variation of
MAPs within the same groups of larvae as a consequence
of the progressive increase of prey availability (see below
for statistical analysis). This design was adopted to
simulate the actual situation occurring in the rearing
mesocosm tanks, that is constant prey concentration with
periodic increase of prey availability. In these tanks, a
more or less constant density of Artemia nauplii was
maintained, by adding nauplii into the tanks every 4 h
during daytime. The density of the rearing tank was taken
as d2 in our experiments.
Since the prey density in the rearing tanks changed in
relation to the larval age, the quantity of nauplii which
had to be added in each replicate, was calculated at the
on
time of the experiments for each experimental group (i.e.
age class). This resulted in the following densities,
measured as number of nauplii/l: d0 = 17.5, d1 = 35 and
d2 = 70 for the 10 days old larvae, d0 = 25, d1 = 50 and
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d2 = 100 for the 20 days old larvae, and d0 = 42, d1 = 84
and d2 = 168 for the 30 days old larvae. Experimental
trials were conducted from 10 March to 11 April 2003.
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The time at which the trials were run were randomised
with respect to the time at which the nauplii were added
to the rearing tanks. The average amount of time passed
from the last addition of nauplii in the rearing tanks to
each experimental trial was statistically compared across
the three age groups, to control for the feeding histories
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of the experimental larvae. With the differences across
experimental groups not statistically significant (143.13 ±
58.28 min for the 10 days group, 123.20 ± 78.85 min
for the 20 days old group, 116.60 ± 53.96 min for the
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30 days group, Kruskal–Wallis, n = 28, P N 0.05), feeding
histories of experimental larvae could be considered
th
similar across treatments. Therefore, it was assumed that
satiation/hunger factors acted with an average similar
intensity on the three experimental groups.
Au
Each experimental trial was conducted as follows: 20
larvae were randomly captured from the original rearing
tank, introduced into the experimental tank, and allowed
to acclimatise for a period of 20 min. Preliminary trials
showed that this time was enough to allow larvae to
recover from stress due to transfer, and assume their
normal behaviour.
Before starting each experimental trial, a digital
videocamera was positioned in front of the experimental
tank. The larval behaviour was then recorded during
three consecutive periods, corresponding to the three
progressive increases in prey availability. Each period
lasted 20 min, during which the behaviour of 4 different
individuals was recorded consecutively for 5 min each
(focal animal sampling, Altman, 1974) and with careful
attention an individual was observed only once during
an observation period (Brown, 1986). At the end of
every 20 min period, the prey density was increased by
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adding nauplii into the tanks by means of a glass-pipette.
At the end of every experimental trial, larvae sub-
samples (n = 4, 75 larvae in the 10 days old group; n = 5,
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97 larvae for 20 days; and n = 5, 76 larvae for 30 days)
were taken, anesthetised using MS-222 and fixed in
neutral formalin 8% for subsequent analysis. Total
length of each of these larvae was measured by means of
a calliper, being respectively 7.03 ± 0.6 mm in the
10 days group, 12.08 ± 1.01 mm in the 20 days group,
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and 14.78 ± 0.73 mm in the 30 days age group. Mean
body size differed significantly across the three groups
(Kruskal–Wallis, n = 248, P b 0.001).
These larvae were dissected and examined under
optical stereo-microscope (Zeiss Stemi SV6 (8×–50×);
Zeiss Standard 20 (32×–100×) to determine the gut
content in terms of the number of Artemia nauplii for
each sea bass larva. The number of individual nauplii
was estimated on the basis of the naupliar carapaces
found in each stomach. The analysis of gut content was
not performed to quantitatively assess prey consump-
tion, rather to control for possible general differences in
feeding tactics and rates across the three age groups. The
eventual presence of other type of preys in the stomach,
besides Artemia nauplii, was also noted.
2.3. Behavioural variables and data analysis
A preliminary analysis of videotapes was carried out
in order to identify the main larval behavioural units
(larval MAPs — Modal Action Patterns — according to
Brown, 1986), which were categorised either as events,
i.e. MAPs of very short duration (usually less than 2 s),
or as activities, when they had a measurable duration.
The quantitative analysis of the behavioural sequences
was then carried out by means of a software package
developed by the authors. This software allowed the
acquisition of temporal sequences of larval MAPs from
the videotapes and their quantification either in terms of
frequency (events/minute) or duration (% time destined
to a given activity with respect to the total recorded
time). Frequency or duration of each MAP was obtained
for each focal animal and then averaged on the 4 focal
animals recorded within each 20 min period of
recording, corresponding to a given prey density. In
 V. Georgalas et al. / Aquaculture 264 (2007) 418–427 421

this way, a single value of frequency or duration of a
given MAP was obtained for each replicate, that is for
each group of larvae with a given age, at a given prey
density. Furthermore, a quantitative analysis of beha-
vioural bouts of each activity was performed. The bout
duration (i.e., the time between the start and the end of a
single occurrence of a behavioral activity, in minutes)
and the bout frequency (number of bouts/min) were
by foraging events. The detailed analysis of the
videotaped behavioural sequences allowed the identifi-
cation of six MAPs, three of which were classified as
activities, and three as events, concerning both swim-
ming and feeding behaviour.
The tree identified “activities” were:
1. Normal swimming (NSw): according to the defini-

py
obtained from the analysis of the behavioural sequences tions given by Rosenthal and Hempel (1970), normal
and then treated with the same approach described swimming was the “ordinary” swimming activity,
above for the MAPs frequency and duration. performed with apparently constant speed and

Means of behavioural variables were then compared
across the three age groups (10, 20, 30 days) within a
given prey density (d0, d1 and d2) by means of
Kruskal–Wallis test (followed by Multiple Comparison
Test; Siegel and Castellan, 1992), and across prey
densities within a given age, by means of Friedman
co
sustained by rapid movements of the caudal fin.
2. Sinking (Sk): when the larva ceases momentarily
“normal swimming” and sinks passively through the
water column.
3. Resting (Rs): when the larva ceases all the activities
and rests immobile on the tank bottom.

ANOVA (Siegel and Castellan, 1992). The Friedman
ANOVA was used to test for behavioural differences
within the same groups of larvae as response towards an
on
increasing level of prey availability; being this test
thought, according to Siegel and Castellan (1992), for
dependent samples that is when the same individuals are
subjected to different conditions.
rs
al
The three “events” were:
1. Abrupt swimming (ASw): according to Rosenthal
and Hempel (1970), this type of swimming consisted
of movements of very short duration often connected
with a shift in orientation.

The number of Artemia nauplii/individual was aver- 2. Sigmoid-posture (SPs): this is an aiming posture, as a
aged on the total number of fish larvae of each replicate part of feeding behaviour, which has been observed
and then compared across the three age groups by means as a typical MAP also in other fish larvae (Munk and
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of the Kruskal–Wallis test. Within each age class, the
number of Artemia nauplii/individual was related to the
individual body size of the larvae by means of the
Spearman Correlation Test (Siegel and Castellan, 1992).
3. Results
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Kiorbe, 1985; Temple et al., 2004). The larval body
assumes an S-curve which could precedes a lunge
and the prey ingestion.
3. Attack (Ak): it consisted of the lunge often
performed subsequently to the Sigmoid-posture,
and it was in many cases followed by movements

associated to prey mastication and ingestion.

3.1. Larval ethogram
As shown in Table 1, the most represented activity in
The larval behaviour consisted of alternating bouts of the larval time budget is the “normal swimming”, to
o

swims and motionless periods, punctuated occasionally which the larvae dedicated more than 60% of the total
th

Table 1
Mean values and range (in parenthesis) of the different MAPs in relation to age (DPH, days post-hatching) and Artemia density
Age Artemia % Time Events/min
Au

(DPH) density
NSw Sk Rs SPs Ak ASw
10 d0 70.44 (57.99–81.80) 25.20 (16.63–29.74) 4.37 (0–19.14) 2.74 (0.68–8.63) 5.11 (3.11–10.99) 0.58 (0–1.88)
d1 72.39 (65.87–76.96) 24.17 (16.53–34.12) 3.44 (0–6.50) 2.46 (1.25–5.97) 4.87 (1.78–9.64) 0.60 (0.11–1.53)
d2 69.06 (63.98–74.55) 27.51 (16.56–32.94) 3.43 (0–8.88) 1.13 (0.34–2.47) 2.80 (1.07–3.86) 0.44 (0.05–1.03)
20 d0 81.23 (71.33–87.55) 15.74 (11.85–26.66) 3.03 (0–4.15) 0.64 (0.10–1.43) 1.95 (0.68–3.89) 0.41 (0.10–0.85)
d1 81.57 (72.61–88.34) 15.97 (11.08–26.32) 2.46 (0–6.84) 0.69 (0.08–1.54) 1.75 (0.11–6.67) 0.49 (0–1.22)
d2 83.91 (70.97–90.93) 15.09 (5.69–29.02) 1.00 (0–7.70) 0.78 (0.16–1.05) 1.40 (0.52–4.36) 0.37 (0–0.66)
30 d0 81.45 (69.05–94.04) 15.06 (5.76–22.43) 3.49 (0–17.41) 0.33 (0–1.22) 1.23 (0–4.91) 0.45 (0.09–1.33)
d1 80.56 (73.48–89.08) 18.92 (10.91–24.68) 0.53 (0–3.20) 0.82 (0.09–2.52) 1.06 (0.11–3.58) 0.35 (0–0.89)
d2 84.47 (77.92–90.68) 15.45 (9.31–21.81) 0.08 (0–0.78) 0.87 (0.09–2.12) 1.03 (0.19–2.23) 0.47 (0.09–0.99)
NSw: normal swimming; Sk: sinking; Rs: resting; SPs: Sigmoid-posture; Ak: attack; Asw: abrupt swimming.
422 V. Georgalas et al. / Aquaculture 264 (2007) 418–427

amount of time. The second activity, in terms of relative 10 days old group; Friedman ANOVA, n = 10, P N 0.05
importance in the time budget, is “sinking”, which was all tests for the 20 and 30 days groups). Overall, these
comprised between 15 and 25%, followed by “resting” results on the patterns of changes in swimming activity
which was always above the 5% of the total time.
Regarding the events, “attack” was the most frequent
one, being on average comprised between 1 and 5
attacks/minutes, followed by Sigmoid-posture (mean
range 0.33–2.74) and, finally, by “abrupt swimming”

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(mean range 0.3–0.6).

3.2. Behavioural variation with age and food density

co
Mean time percent of “normal swimming” and
“sinking” changed significantly with age, but not with
food density (Kruskal–Wallis, n = 28, P b 0.05, all tests
and Friedman ANOVA, n = 8, P N 0.05 all tests for the
10 days old group; Friedman ANOVA, n = 10, P N 0.05

al
all tests for the 20 and 30 days groups). The two
activities showed complementary patterns, with the
“normal swimming” increasing while “sinking” de-
on
creasing with age (see Fig. 1a and b). Both activities
changed significantly between 10 and 20 days but not
between 20 and 30 days (Kruskal–Wallis, Multiple
Comparison Test, P b 0.05). At d1 and d2 density, mean
rs

time percent of “resting” decreased significantly with

age whereas at density d0 there was no statistically
significant difference across ages (Kruskal–Wallis,
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n = 28, P N 0.05). In the 30 days age group, the time

percent of this activity and its variation decreased
significantly to zero with increasing food density
(Friedman ANOVA, n = 10, P b 0.05, Fig. 1c).
Results of bout analysis showed very similar
patterns: the mean duration of “normal swimming”
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bouts increased significantly with age (Kruskal–Wallis,

n = 28, P b 0.05, Table 2), whereas the number of bouts/
min of this activity decreased, with main changes
occurring also in this case between 10 and 20 days post-
o

hatching. Mean duration of bouts of “sinking” increased

slightly across age groups, but their frequency decreased
th

significantly from 10 to 20 days post-hatching (Krus-

kal–Wallis, n = 28, P b 0.05, Table 2). A similar pattern
was detected in the mean duration and frequency of
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“resting” bouts, and in this case a strong effect of food

density was also present: as shown for the mean time
percent, mean duration of these bouts decreased
significantly in the older larvae as food density
increased, especially in the 30 days age group (Friedman
ANOVA, n = 10, P b 0.05, Table 2).
Mean frequency of abrupt swimming did not change
Fig. 1. Mean ± S.E. of activity (% time) at each age and prey density: a)
significantly neither with age, nor with food density NSw; normal swimming, b) Sk; sinking, c) Rs; resting. Values are the
within age (Kruskal–Wallis, n = 28, P N 0.05, all tests, means of eight replicates for the 10 days old group and ten replicates
Friedman ANOVA, n = 8, P N 0.05 all tests for the for the 20 and 30 days old group respectively.
 V. Georgalas et al. / Aquaculture 264 (2007) 418–427 423

(0.000–3.534)
(0.000–1.650)
(0.000–3.486)

(0.000–1.774)
(0.000–1.860)
(0.000–1.319)
(0.000–0.794)
(0.000–0.297)
(0.000–1.112)
showed that at older age and, in some cases, at higher
food density, larvae became more active, with the non-
swimming activities decreasing in terms of both
duration and frequency.

1.115
1.251

1.066
0.471
0.672
0.277
0.499
0.170
0.030
Frequency of Sigmoid-postures decreased signifi-

Rs
cantly from a mean higher than 1 postures/min in the
10 days old larvae to a mean lower than 1 in the 20 and

(8.483–13.071)
(8.459–13.613)

(3.475–12.474)
(3.729–12.300)

(3.831–19.229)
(8.562–11.873)

(5.231–11.193)
30 days old larvae, as regards to the only d0 and d1 food

py
(2.209–7.360)

(2.864–8.552)
densities (Kruskal–Wallis, n = 28, P b 0.01 both tests;
Fig. 2a). The frequency of attacks decreased similarly
from 10 days age to 20 days age, and this difference was

co
statistically significant at each food density (Kruskal–

10.274
10.194
11.587
6.692
5.848
6.283
5.350
7.438
4.915
Wallis, n = 28, P b 0.01 all tests; Fig. 2b).

Sk
Frequency of both attacks and S-postures changed
also significantly across food densities, but pattern of
variation depended on the particular age. Within 10 days

12.464 (10.469–13.928)

13.380 (12.456–14.424)
12.343 (9.154–15.754)

7.103 (4.972–13.033)
7.127 (4.246–12.967)

8.371 (4.363–21.921)
7.809 (5.881–11.797)

6.354 (2.931–8.955)

5.535 (3.616–9.348)
old larvae, frequency of S-postures and attacks decreased
with food density (Friedman ANOVA, n = 8, P b 0.05

al Mean values and range (in parenthesis) of the different MAPs in relation to age (DPH, days post-hatching) and Artemia density
both tests), whereas in 30 days old larvae, frequency of

Bouts/min
S-postures increased with increasing food density,
on
especially from d0 to d1 (Friedman ANOVA, n = 8,

NSw
P N 0.05 all tests for the 10 days old group; Friedman
ANOVA, n = 10, P N 0.05 all tests for the 20 and 30 days
groups; Fig. 2a, b).
rs

0.011 (0.000–0.038)
0.009 (0.000–0.018)
0.014 (0.000–0.043)
0.019 (0.000–0.054)
0.010 (0.000–0.049)
0.004 (0.000–0.017)
0.019 (0.000–0.078)
0.004 (0.000–0.023)
0.001 (0.000–0.005)
Finally, the ratio between the frequency of S-posture
and that of attacks was calculated for each replicate, as a
measure of the degree to which the two behaviours were
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associated. Patterns of variation appeared to depend on

the interaction between age and food density. This index
Rs

decreased significantly with age, only within the d0

food treatment (Kruskal–Wallis, n = 28, P b 0.05 all
0.024 (0.016–0.030)
0.026 (0.018–0.047)
0.024 (0.020–0.028)
0.025 (0.016–0.051)
0.029 (0.017–0.046)
0.028 (0.012–0.067)
0.030 (0.016–0.065)

0.034 (0.019–0.045)
0.032 (0.011–0.050)
tests, Fig. 2c), whereas it increased significantly with
age in both d1 and d2 treatment (Kruskal–Wallis,
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n = 28, P b 0.05). Within 20 and 30 days age, the index

increased strongly with food density, especially from d0
to d1 treatment, as shown in Fig. 2c for the 30 days old
Sk

larvae. Within the 30 days old larvae, the index is close

o

to 1 whereas in the younger larvae the ratio is less than 1,

indicating that a higher number of attacks were
NSw: normal swimming; Sk: sinking; Rs: resting.
th

0.086 (0.043–0.221)
0.064 (0.048–0.080)
0.057 (0.049–0.068)
0.150 (0.091–0.407)
0.162 (0.083–0.347)
0.179 (0.100–0.489)
0.212 (0.093–0.484)
0.135 (0.049–0.229)
0.194 (0.084–0.270)
Bout duration (min)

performed with respect to the Sigmoid-postures. An

equal frequency of attacks and postures in the older age
indicates that these two behaviours are more strictly
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associated in these larvae than in the younger larvae.

NSw

The mean number of ingested Artemia nauplii/

individual increased progressively from a value of 2.6
in the 10 days old larvae, to 20.5 in the 20 days old
Artemia
density

larvae and, finally, to 58 in the 30 days old larvae

(Fig. 3). Mean values differed significantly across the
d0
d1
d2
d0
d1
d2
d0
d1
d2

three age groups (Kruskal–Wallis, n = 13, P b 0.01). The

mean number of individuals with empty stomach
Table 2

(DPH)

decreased with age but the difference was not fully

Age

10

20

30

significant (Kruskal–Wallis, n = 13, P = 0.07; Fig. 3).

424 V. Georgalas et al. / Aquaculture 264 (2007) 418–427

larvae (Spearman Correlation, r 2 = 0.38, n = 97,

P b 0.05; r2 = 0.42, n = 76, P b 0.05 for the 20 days and
30 days old larvae respectively), but not within the
10 days old larvae (Spearman Correlation, r2 = 0.055,
n = 75, P N 0.05).
Rotifers were also occasionally found in the stomach
of 10 days old larvae, whereas in the other two age
classes only Artemia nauplii were found as prey.

py
4. Discussion

co
The results of the present study showed the
ontogenetic changes in the behaviour of European sea
bass larvae. A clear result is the significant increase of
swimming activity, especially from 10 to 20 days post-
hatching. As found out by Diaz et al. (2003), the beha-
vioural changes in the larval phase are paralleled by the

al
progressive formation of a more efficient sensory
system.
In particular, our results showed that, as the
on
swimming activity increased significantly from 10 to
20 days, the time spent sinking and resting decreased.
Similar ontogenetic changes have been also reported for
other Teleost fishes (Brown, 1986; Pittman et al., 1990;
rs

Munk, 1995; Rabe and Brown, 2001; Utne-Palm and

Stiansen, 2002). This should coincide with a remarkable
increase in the efficiency of behaviours related to prey
pe

searching and capture, as it will be discussed later, and

this could be also paralleled, at the morpho-physiolog-
ical level, by a full development of the swim bladder.
Our results are fully consistent with what has been found
o r's
th
Au

Fig. 2. Mean ± S.E. of events per minute for each age and prey density: a)
SPs; S-posture, b) Ak; attack; and c) mean ± S.E. of the ratio between SPs/
Ak events. Values are the means of eight replicates for the 10 days old
group and ten replicates for the 20 and 30 days old group respectively.

Fig. 3. Mean ± S.E. of Artemia nauplii per individual/replicate (left

Larval body size and number of ingested nauplii were axis) and number of individuals with empty gut (right axis) for the
positively related, but this relationship was statistically three different ages. Values are the means of 4 replicates in the 10 and
significant only within the 20 days and the 30 days old 30 days old group and 5 replicates in the 20 days old group.
 V. Georgalas et al. / Aquaculture 264 (2007) 418–427
by Chatain (1986) regarding the swim bladder devel-
opment in the sea bass: this author reports an initial
phase of inflation around the 7th day followed by an
“expansion phase” when the larva reaches 12 mm of
body length. This stage coincides with the 20 days old
group of larvae used in the present study. These findings
suggest that all the major changes in the larval swimm-
ing behaviour, reported by the present study, could be
strictly related to the swim bladder development. The
complete swim bladder development should improve
the general swimming performance of the larvae, by
allowing neutral buoyancy.
Despite these differences between age classes, the
swimming activity (normal swimming) in the sea bass
larvae is in every case rather intense, representing
always more than 60% of the total time of activity. If
these results are compared with that found in larval cod
and herring, as reported by Munk and Kiorbe (1985) and
Munk (1995), it is possible to conclude that larval sea
bass shows a behavioural tactic more similar to that of
on
herring larvae with respect to that shown by the cod.
Larval sea bass appeared to fall into the category of
“cruise predators”, showing a more continuous and
intense swimming, while cod showed a so called
rs
“saltatory strategy” (Munk, 1995), characterised by
more frequent pauses and reduced swimming activity.
Further, our results on duration and frequency of bouts
pe
of activities clearly indicate that there is an ontogenetic
shift of the behavioural tactic from “saltatory” to
“cruising”, as shown by Rabe and Brown (2001) in
the witch flounder. The swimming activity seems to
become significantly more continuous as the larva
grows, with the major shift occurring from 10 to 20 days
r's
post-hatching.
Nonetheless, the present results differ from that
obtained by Munk and Kiorbe (1985) and Munk
(1995) on herring and cod regarding the relationship
o
between swimming activity and prey density, although
these studies are not fully comparable with the present
th
one, due to some differences in the experimental de-
sign. In particular, in our experiments no water tur-
bulence was created in the experimental tank, and this
Au
factor could strongly affect the larval behaviour in
many respects, such as the encounter rate between
preys and larvae.
Results of the above cited authors clearly indicated
that swimming activity decreased linearly with prey
density, whereas in the present work such pattern has not
been found. Rather, we found that, especially at higher
densities, time destined to resting decreased with age
and, within older larvae, this variable decreased also
with increasing food density.
425
This result seems to further indicate that sea bass
larvae adopt a “cruise” predatory strategy, leading to an
increase of the swimming activity with food density.
Variation in the frequency of feeding behaviours
appeared to be affected by both age and food density.
Regarding the effect of age, our results contrast
apparently with that found by Rabe and Brown (2001)
and Utne-Palm and Stiansen (2002), since these authors
py
found an increase of the attack rate with age in herring
larvae, whereas the present work showed a tendency to
decrease of both attacks and S-postures frequency with
co
age. However, our results indicate that these two beha-
viours are performed with a ratio of 1 to 1 and therefore
they are associated within a behavioural sequence, only
in the older larvae and at higher food density. This could
indicate that feeding behaviour is fully efficient only
when the S-posture is followed by an attack and that
al
attacks not preceded by an S-posture, are probably
misdirected or unsuccessful behaviours.
In younger larvae (especially the 10 days old
larvae), the number of attacks is higher than the
number of Sigmoid-postures and both behaviours are
not positively related with food density, indicating that
they are not efficient in terms of prey capture and es-
pecially in terms of ratio between attempts and suc-
cessful capture. In other words, only the complete
behavioural sequence composed by an aiming posture
and an attack should be considered as a MAP with a
full feeding function.
The frequency of this MAP seems to properly develop
with age, being modulated by food density, only within
the older larvae, and this is fully in agreement with
previous studies on the relationship between feeding
behaviour, larval age and prey density (Munk, 1995; Cox
and Pankhurst, 2000; Rabe and Brown, 2001). An
alternative explanation could be related to satiation
processes which may act in the younger larvae but not in
the older ones: this would explain the decrease in feeding
MAPs observed in the 10 days larvae in relation to the
increase of prey availability.
An inverse relationship between feeding efficiency
and age is also confirmed by the analysis of the gut
content of the investigated larvae, as the number of
Artemia nauplii/individual strongly increased with age,
whereas the number of individuals found to have their
gut empty tended to decrease. Results on gut content
assessed after the experiments could account not only
for the larval behaviour observed in the experimental
tanks, but also for the feeding behaviour previously
adopted by the larvae in the rearing tanks. However,
these results indicated an ontogenetic difference, which
seemed to parallel the behavioural data.
426 V. Georgalas et al. / Aquaculture 264 (2007) 418–427
These results could possibly be attributed to the
change in the ratio between the prey body size and the
fish larval body size occurring with the growth of the
fish larva: 10 days old larvae are still fed also with
rotifers, and their feeding ability on Artemia nauplii
could be limited just by their body size according to
Diaz et al. (2003). By contrast, as body size increases,
the ability to ingest a larger number of nauplii could
increase with a disproportional pattern, allowing the
larvae both to capture more efficiently a larger number
of preys, and to adjust their behaviour to food density.
According to the available literature (Cunha and Planas,
1999; Cox and Pankhurst, 2000), this pattern could be
explained by the increase of the mouth size and the
parallel decrease of the duration of the feeding event for
a given prey size with the increasing of the larval body
size.
This is also consistent with the positive relationship
between larval body size and the number of ingested
nauplii, observed within the 20 and the 30 days old
on
groups. The positive relationship between the number of
preys and body size indicates a size dependent
mechanism of prey capture, due to the higher capture
success of larger larvae. The lack of this relationship in
rs
the 10 days old larvae suggests that the level of inter-
individual variation in body size is too low, within these
young larvae, to generate body-size related differences.
pe
However, the presence of a relatively large number of
individuals with empty stomach in this group suggests
that a certain degree of inter-individual variation in the
prey capture ability could also act at this age.
The behavioural ontogenetic sequence, as reported
for sea bass larvae in the present study, could be used in
r's
future research for comparisons with that obtained for
larvae reared in conditions different from the mesocosm
technique. Larval ethogram could be therefore regarded
as a tool for the assessment of the quality of the reared
o
fish, which could be combined with the morphological
and genetic tools.
th
Acknowledgments
Au
We are grateful to Professor Stefano Cataudella and
Dr. Attilio Spanò for having provided the hatchery and
experimental facilities for the present study. We would
also like to thank Mr. Piero Trebbi for the technical
support given to the experimental set-up.
This study was funded by the Italian Ministry of
Agricultural Resources (M.I.P.A.F, VI Piano triennale
della pesca e dell'acquacoltura: “Ripopolamento attivo
di lagune, stagni costieri, e localizzate aree della fascia
costiera con giovanili certificati di specie ittiche e di
crostacei, secondo i principi del codice di condotta per
una pesca responsabile (FAO, 1995)”).
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