JOURNAL OF EXPERIMENTAL ZOOLOGY 292:573–579 (2002)

Temperature Sex Determination in the European

Sea Bass, Dicentrarchus labrax (L., 1758) (Teleostei,
Perciformes, Moronidae): Critical Sensitive
Ontogenetic Phase
GEORGE KOUMOUNDOUROS,1,2* MICHALIS PAVLIDIS,1,2
LINA ANEZAKI,1 CONSTANTINA KOKKARI,1 ASPASIA STERIOTI,1
PASCAL DIVANACH,1 AND MAROUDIO KENTOURI1,2
1
Institute of Marine Biology of Crete, GR-71 003, Heraklion, Crete, Greece
2
University of Crete, Department of Biology, GR-71 409, Heraklion, Crete,
Greece

ABSTRACT The temperature sex determination (TSD) mechanism in the European sea bass
(Dicentrarchus labrax L.) was studied in respect to: a) the TSD sensitivity during the different
developmental stages; and b) the intrapopulation correlation of sex determination with the growth
rate up to the end of the TSD-sensitive period. At the stage of half-epiboly, eggs from the same
batch were divided into four groups and subjected to different thermal treatments: a) 15°C (G15
group) and b) 20°C (G20 group) up to the middle of metamorphosis stage; c) 15°C up to the end of
yolk-sac larval stage and subsequently to 20°C (G15-5 group); and d) 15°C up to the end of the
preflexion stage and then to 20°C (G15-10 group). At the end of the treatments, size grading was
applied and four additional populations were established from the upper (L) and lower (S) size
portions of the G15 and G20 populations: G15L, G15S, G20L, and G20S. During the following
growing phase, all populations were subjected to common rearing conditions. The sex ratios of
each population were macroscopically determined at 190–210 mm mean total length. Female inci-
dence was significantly affected (P < 0.05) by the different thermal treatments: 66.1% in the G15,
47.1% in the G15-10, 37.6% in the G15-5, and 18.1% in the G20 group. In addition, sex ratio was
correlated with the growth rate of the fish up to the end of the TSD-sensitive period, with the
larger fish presenting a significantly higher (P < 0.01) female incidence than the smaller fish in
both thermal regimes tested: 73.1% in G15L vs. 57% in G15S, and 36.6% in G20L vs. 22.5% in
G20S group. Results provide, for the first time, clear evidence that the sea bass is sensitive to
TSD during all different ontogenetic stages up to metamorphosis, and that sex ratio is correlated
with the growth rate of the fish well before the differentiation and maturation of the gonads. J.
Exp. Zool. 292:573–579, 2002. © 2002 Wiley-Liss, Inc.

Phenotypic sex in several gonochoristic fish is
considered to be the outcome of genetic and envi-
ronmental or social (population density, relative
size of juveniles) factors. Among the several envi-
ronmental factors studied so far, the thermal con-
ditions prevailing during the period of primary sex
differentiation appear to be the main environmen-
tal determinant of sex (reviewed by Baroiller et
al., ’99). The phenomenon of temperature sex de-
termination (TSD) in fish was first indicated in
Menidia menidia, where female or male biased
populations were reported to follow exposure to
high or low temperatures, respectively (Conover
and Kynard, ’81). Since then, TSD has presented
in several fish species, such as Odontesthes bon-
ariensis (Strüssmann et al., ’96a), O. argentinensis
(Strüssmann et al., ’96b), Poecilia melanogaster and
Apistogramma spp. (Römer and Beisenherz, ’96),
Patagonina hatcheri (Strüssmann et al., ’97),
Oreochromis niloticus (Baroiller et al., ’95; Baras
et al., 2001), O. aureus (Desprez and Mélard, ’98;
Baras et al., 2000), Paralichthys olivaceus (Yama-
moto, ’99), O. mossambicus (Wang and Tsai, 2000),
and Dicentrarchus labrax (Pavlidis et al., 2000),
with a great range of response to the respective
temperature effect.
*Correspondence to: George Koumoundouros, University of Crete,
Department of Biology, P.O. Box 2208, GR-71 409, Heraklion, Crete,
Greece. E-mail: koumound@imbc.gr
Received 6 August 2001; Accepted 10 January 2002
Published online in Wiley InterScience (www.interscience.wiley.com).
DOI: 10.1002/jez.10095

© 2002 WILEY-LISS, INC.

574 G. KOUMOUNDOUROS ET AL.
Thermolabile sex determination has a great
adaptive significance for wild fish populations
(Conover and Heins, ’87a,b; Strüssmann et al.,
’96a,b; Baras et al., 2000) and a high practical
interest for aquaculture (reviewed by Baroiller et
al., ’99; Beardmore et al., 2001). Furthermore, the
identification of the TSD-labile ontogenetic period
is of great importance for wild fish, since it deter-
mines the sex ratio of the natural populations in
relation to the different environmental preferences
of the various developmental stages. Besides, it
could elucidate the exact critical period of TSD in
reared fish, to avoid the negative side effects of
potentially harmful temperature conditions. Pre-
vious studies suggested that the critical period of
TSD in fish is located just prior to the histologi-
cally recognizable gonadal differentiation (Conover
and Fleisher, ’86; Strüssmann et al., ’97) or dur-
ing the sensitive period for exogenous steroid-in-
ducible sex determination (Strüssmann et al., ’96a;
Blàzquez et al., ’98; Baroiller et al., ’99).
The European sea bass, D. labrax, is a gono-
choristic species that lacks heteromorphic sex
chromosomes (Cataudella et al., ’73; Sola et al.,
’93). The first primordial germ cells of the gonads
appear at 10.6 mm standard length (SL), but go-
nads become differentiated only after 90–120 mm
SL (Roblin and Bruslé, ’83; Blàzquez et al., ’99).
Recently, it has been shown (Pavlidis et al., 2000)
that the sea bass presents a moderate to strong
TSD during early ontogeny (stage of half-epiboly
to metamorphosing larvae of 17–18 mm total
length), i.e., well before the critical period of sen-
sitivity to exogenous steroids (20 to 40–80 mm
fork length and 57–137 days post-fertilization, af-
ter Blàzquez et al., ’98). Although relatively small,
the defined temperature sensitive period for sex
determination in sea bass, as well as in other fish
species, is composed by different developmental
stages with clearly different morphological and
allometric patterns, which reflect significant func-
tional changes (Koumoundouros et al., 2001).
This study will explore the critical sensitive on-
togenetic phase of TSD in sea bass during the dif-
ferent developmental stages up to 17–18 mm total
length (TL). In addition, to elucidate the higher
growth rate observed in female fish (Pavlidis et al.,
2000), the intrapopulation correlation of sex deter-
mination with the growth rate will be investigated.
MATERIALS AND METHODS
Eggs were obtained (February 16, 2000) from a
single batch of a wild-caught broodstock, sponta-
neously matured and spawned. Broodstock matu-
ration and spawning, as well as larval and juve-
nile rearing, were performed according to Pavlidis
et al. (2000). All the established experimental
populations were in duplicate.
TSD-sensitive ontogenetic period
The response of sea bass to the TSD during dif-
ferent developmental phases was examined at two
temperatures, one female inducing (15°C) and one
male inducing (20°C) (Pavlidis et al., 2000). Thus,
fish ontogeny was performed (Fig. 1) at: a) 15°C
(Group G15) or at b) 20°C (Group G20) during
the whole thermolabile developmental period
(half-epiboly to middle of metamorphosis phase)
(Pavlidis et al., 2000), and at c) 15°C up to the
end of yolk-sac larval stage and subsequently at
20°C (Group G15-5), or d) at 15°C up to the end
of the preflexion stage (all the fish presented a
flexed notochord) (Koumoundouros et al., 2001)
and then at 20°C (Group G15-10). At the end of
the different thermal treatments, all fish were
subjected to a salinity floating test, to remove the
individuals without an inflated swimbladder
(Chatain and Corrao, ’92) and then were counted.
Afterward, a random sample of 500 individuals
was taken from each experimental population and
subjected to common rearing conditions (Pavlidis
et al., 2000).
Relationship of body size at the end of the
TSD-sensitive period with sex determination
Following the salinity floating test and a re-
covery period of two weeks, size grading was ap-
Fig. 1. Experimental design followed, in respect to the de-
velopmental stage, fish size, and age. Thermal treatment (G15
= 15°C; G15-10 = 15°C up to the end of the preflexion stage
and subsequently at 20°C; G15-5 = 15°C up to the end of
yolk-sac larval stage and subsequently at 20°C; G20 = 20°C)
lasted from the stage of half-epiboly until the middle of meta-
morphosis, and then fish were exposed to common rearing
conditions. Solid heavy lines: exposure of fish to 15°C; Dashed
line: exposure of fish to 20°C; Solid light line: exposure of
fish to common rearing conditions; TL: total length; PHD:
post-hatching days; TSD: temperature sex determination.
 ONTOGENETIC SENSITIVITY OF TSD IN SEA BASS
plied to select the upper and lower size portions
(5–10%) of the populations reared at constant
temperatures (15°C and 20°C) during the TSD-
sensitive period. Size grading was based on the
frequency distribution of the individual body
weight of a random sample of 100 fish (Fig. 2),
taken from each population following anaestheti-
zation (Ethylenglycol-monophenylether, Merck,
München, Germany, 0.2–0.5 ml/l–1) and weighing
(± 1 mg). Subsequently, a random sample of 300
individuals was taken from each size class (small
and large fish) and experimental groups (G15 and
G20) and subjected to common rearing conditions.
Size grading resulted in significant (P < 0.001)
differences of the body weight between the small
(S) and large (L) population classes of G15 and
G20 groups (Table 1).
Sampling and sex identification
Sex identification was carried out on 47–58 sea
bass juveniles, randomly sampled from each popu-
lation at 190–210 mm mean TL (300–342 days
post TSD-sensitive period, Fig. 1). The gonads of
anaesthetized, measured (± 1 mm TL), and
weighed (± 0.1 g) fish were dissected, and three
Fig. 2. Frequency distribution of the body weight, prior
to size grading, of the populations reared at 15°C (G15) or
20°C (G20) from the stage of half-epiboly until the middle of
metamorphosis. W: body weight.
575
TABLE 1. Mean weight (mg) of the different subpopulations,
following the size grading1
Mean W SD Min Max n
G15S 495a 43 426 550 20
G15L 1316a 138 1208 1572 20
G20S 510b 33 420 561 20
G20L 1281b 118 1149 1350 20
1
Size grading was performed at the end of the different thermal treat-
ments, following a two-week acclimation period. S or L, small or big
size class, respectively, of the populations reared at 15 (G15) or 20°C.
W, body weight; SD, standard deviation; n, sample size. Thermal treat-
ments lasted from the stage of half epiboly to the middle of metamor-
phosis, and then fish were exposed to common rearing conditions.
Same superscript letters show statistical differences between groups
(P < 0.001).
independent investigators macroscopically deter-
mined sex. Macroscopically undifferentiated go-
nads (< 10%) were histologically examined for sex
determination (Pavlidis et al., 2000).
Data management and statistical analysis
The effect of temperature was evaluated by cal-
culating thermal sums (product of temperature
and age) for each population (Fuiman et al., ’98).
As 15°C was shown to be the lower temperature
threshold of TSD in sea bass (Pavlidis et al., 2000),
thermal sums were estimated by using the differ-
ence of each temperature regime from 15°C.
The student’s t-test or Mann-Whitney U-test,
in the case of not normally distributed data or
heterogeneous variances, was applied to check any
significant differences between the different ex-
perimental groups in the body weight and TL. Sur-
vival and sex proportion among the different
experimental groups were compared by the use
of the G-test (Sokal and Rohlf, ’81).
RESULTS
During the embryonic and larval stages, the
different experimental groups presented similar
survival rates (37–47%, P > 0.05). During the fol-
lowing experimental period, survival rate was
high and the same for all groups (92–96%).
The rearing temperature significantly affected
the sex ratio in the experimental populations,
with the G15 and G20 groups presenting the
highest (66.1%) and lowest (18.1%) female inci-
dence, respectively (Table 2). In the experimen-
tal groups G15-5 and G15-10, as the duration of
exposure at 20°C was increased, the lower female
proportion was observed (compared to G15, P <
0.001) without, however, approaching the female
incidence of the G20 group (P < 0.001, Table 2).
The graphical correlation of female incidence on
576 G. KOUMOUNDOUROS ET AL.

TABLE 2. Comparison of body weight, total length smaller fish in both thermal regimes tested (Table
and female incidence between the differently 3). However, the maximum and minimum female
thermally treated groups1
incidence that was presented at the lower (G15)
TL ± SD W ± SD F and upper (G20) range of TSD in sea bass were
Group Sex (mm) (g) n (%) not affected by the growth rate of the individuals;
G15 F 200 ± 15 75 ± 18 72 66.11,2,3 the G15L and the G20S groups were not signifi-
M 200 ± 16 74 ± 19 37 cantly different from the G15 and the G20 groups,
G15-10 F 203 ± 18 79 ± 21 48 47.11,4 respectively (P > 0.05, Table 3). In each respec-
M 200 ± 18 76 ± 21 54 tive experimental population, no significant dif-
G15-5 F 202 ± 14 79 ± 18 41 37.62,4
M 196 ± 20 76 ± 22 68 ferences were observed between the sizes of the
G20 F 191 ± 16 68 ± 15 19 18.11,2,3 two sexes (P > 0.05), except of G20L where males
M 195 ± 19 75 ± 21 86 were significantly larger than females (P < 0.01
1
Thermal treatment (G15 = 15°C; G15-10 = 15°C up to the end of the for TL, P < 0.05 for W) (Table 3).
pre-flexion stage and subsequently at 20°C; G15-5 = 15°C up to the
end of yolk-sac larval stage and subsequently at 20°C; G20 = 20°C) DISCUSSION
lasted from the stage of half epiboly until the middle of metamorpho-
sis, and then fish were exposed to common rearing conditions. TL, In several actinopterygians, as well as in many
total length; W, body weight; n, number of fish; F, females; M, males.
Same superscript numbers indicate significance difference at P < 0.001
reptiles and some amphibians, the sexual pheno-
(1,2,3) or at P < 0.05 (4). type is determined, at least in part, by the incu-
bation temperature of eggs (Pieau et al., ’95;
the thermal sum above 15°C, showed that the pat- Baroiller et al., ’99). The main types of tempera-
tern of decrease of the female incidence was con- ture-dependent sex determination (TSD) among
tinuous for the G15, G15-10, and G15-5 groups, reptiles, amphibians, and fish are: high tempera-
but not for the G20 group, which presented lower tures yield female or male biased populations;
female incidence than what was theoretically ex- both high and low temperatures induce female
pected from the model (Fig. 3). There was no sta- TABLE 3. Comparison of body weight and total length
tistically significant difference in the body size between the different size classes at the end of the
between the two sexes of each experimental popu- experiment.1
lation, at the age of 342 days post TSD sensitive- TL ± SD W ± SD
period (P > 0.05, Table 2). Group Sex (mm) (g) n %F
Sex ratio in sea bass was significantly corre-
G15L F 212 ± 17 1
101 ± 262
79 73.1
lated with the growth rate of fish up to the end of M 211 ± 173 98 ± 254 29
the TSD-sensitive period (expressed as body size G15S F 198 ± 181 82 ± 252 54 57.4
at a specified age). Larger fish showed a signifi- M 197 ± 223 81 ± 304 40
cantly higher (P < 0.01) female incidence than the G20L F 205 ± 19a 99 ± 28b 37 36.6
M 213 ± 185,a 110 ± 316,b 64
G20S F 200 ± 22 91 ± 29 25 22.5
M 201 ± 155 91 ± 216 86
Comparison of female incidence between the different
experimental groups.2

G15L G15S G20 G20L G20S

G15 ns ns *** *** ***

G15L – ** *** *** ***
G15S – *** *** ***
G20 – *** ns
G20L – **
1
Thermal treatment (G15 = 15°C; G20 = 20°C) lasted from the stage
of half epiboly until the middle of metamorphosis, and then fish were
exposed to common rearing conditions. Following a recovery period
Fig. 3. Relationship of female incidence with the water of two weeks after the thermal treatment, size grading was applied
to select the upper (L) and lower size portions (S) of the G15 and G20
temperature and duration of exposure from the stage of half-
groups. TL, total length; W, body weight; n, number of fish; F, fe-
epiboly until the middle of metamorphosis. Thermal sums males; M, males. Same superscripts indicate significance difference
are expressed in days °C above 15°C. G15 = 15°C; G15-10 = at P < 0.001 (1,2), P < 0.01 (a,3), or at P < 0.05 (b,4,5,6).
15°C up to the end of the preflexion stage and subsequently 2
Statistically significant differences between the female incidence of
at 20°C; G15-5 = 15°C up to the end of yolk-sac larval stage the different populations are indicated with *** (P < 0.001), ** (P <
and subsequently at 20°C; G20 = 20°C. 0.01), or * (P < 0.05).
 ONTOGENETIC SENSITIVITY OF TSD IN SEA BASS
populations with males emerging from interme-
diate temperatures, while, in one case, both high
and low temperatures induce monosex male popu-
lations whereas intermediate temperatures yield
a 1:1 sex ratio (Conover and Kynard, ’81; Sullivan
and Schultz, ’86; Conover and Heins, ’87a,b;
Middaugh and Hemmer, ’87; Lagomarsino and
Conover, ’93; Römer and Beisenherz, ’96; Strüss-
mann et al., ’96a,b; Strüssmann et al., ’97; Desprez
and Mélard, ’98; Yamamoto, ’99; Wang and Tsai,
2000; Baras et al., 2000, 2001).
In several fish species, temperature exerts its
effect on sex differentiation only when applied at
a specific ontogenetic phase. In the Atherinid spe-
cies studied so far, the sensitive period of sex
determination is presented during the larval de-
velopment and just prior to the differentiation of
gonads (8–21 mm TL in M. menidia after Conover
and Fleisher, ’86; 11–18 mm TL in O. bonariensis
after Strüssmann et al., ’97), while in the cichlid
Apistogramma trifasciata the sensitivity to TSD
is expressed during the embryonic and the larval
stage (Römer and Beisenherz, ’96).
Recently, it has been shown that, in the Euro-
pean sea bass, higher temperatures favor the for-
mation of males whereas lower temperatures favor
the formation of females (Pavlidis et al., 2000).
The present data are in accordance and provide
more evidence that a thermolabile sex determi-
nation mechanism occurs in sea bass, with a criti-
cal period from the stage of half-epiboly to the
middle of metamorphosis. Additionally, the dura-
tion of thermal manipulation for production of
female-biased populations has been explored. Re-
sults clearly showed that temperature exerts its
effect throughout the tested developmental period,
and that the proportion of males was increased
with increasing thermal sums. However, the com-
parison of female occurrence between the G15,
G15-5, and G20 groups indicated that the early
developmental period up to the end of yolk-sac
larval stage is relatively more sensitive than the
following feeding larval stage, resulting in the
same increase of the female incidence (20% in-
crease from G20 to G15-5 vs. 28% increase from
G15-5 to G15), but in seven folds shorter develop-
mental period (27 vs. 211 degree days above 15°C).
There is a general perception that phenotype
sex in gonochoristic fish can only be manipulated
around the time of the gonadal differentiation pe-
riod. In tilapias, some atherinids, and in reptiles,
the thermosensitive period is similar to the labile
period, i.e., the critical period during which the
gonads are still undifferentiated but exhibit maxi-
577
mum sensitivity to the action of exogenous hor-
monal treatment (Baroiller et al., ’99). In sea bass,
the differentiation of the primordial gonads ap-
pears to occur at 14.0 mm TL (Roblin and Bruslé,
’83), and the labile period is located from 57 to
137 days post fertilization (Blàzquez et al., ’98).
Our results suggest that temperature exerts its
effect on sex determination not only prior to the
differentiation of the primordial gonads, but even
before the histological differentiation of the first
germ cells (9.0 mm TL, Roblin and Bruslé, ’83),
indicating that presumptive gonads are commit-
ted to a certain sex phenotype prior to gonadal
differentiation. Under this assumption, different
incubation temperatures may regulate the expres-
sion of sex-determining genes at hypothalamic
sites and the hypothalamic control of secretion of
several hormones involved in sex differentiation
(Janzen and Paukstis, ’91; Francis, ’92; Pavlidis
et al., 2000).
The European sea bass, as well as other fish
species, exhibit a clear sexual growth dimorphism,
which is expressed well after the sex differentia-
tion (Blàzquez et al., ’98; Yamamoto, ’99; Pavlidis
et al., 2000; Beardmore et al., 2001). This dimor-
phism could be explained by the fact that male
sea bass reach sexual maturation one year ear-
lier than females (Stequert, ’72; Bruslé and Roblin,
’84; Kara, ’97), thus expending relatively more en-
ergy on gonadal growth. In addition to this sexual
dimorphism, the present paper showed that the
growth rate up to the end of the TSD-sensitive
period (i.e., well before sexual differentiation and
maturation) is significantly correlated with sex
determination in the European sea bass, with the
males being favored in the smaller and the fe-
males in the bigger size classes of a unique popu-
lation. However, the correlation of early growth
rate with sex determination is strongly limited in
the range of TSD-induced sex ratio, with the G15
fast growing and the G20 slow growing—individu-
als do not exceed the thermally determined maxi-
mum and minimum female frequency, respectively.
This result, together with the fact that the sex
ratio was not further altered at temperatures
lower than 15°C (Pavlidis et al., 2000), indicates
that, similar to other fish species (reviewed by
Baroiller et al., ’99), the sex plasticity in sea bass
can not lead to monosex populations. Therefore,
although sea bass lack heteromorphic sex chro-
mosomes (Cataudella et al., ’73; Sola et al., ’93),
it can be assumed that sex plasticity in sea bass
is partially controlled by GSD mechanisms. A
physiological or genetic correlation of the growth
578 G. KOUMOUNDOUROS ET AL.
rate with sex determination may provide an ex-
planation for the growth sexual dimorphism ob-
served by the present study at the end of the
TSD-sensitive period. Since all the populations
were subjected to the same rearing conditions and
were fed ad libitum, the side effects of other fac-
tors on the correlation of growth rate with sex
determination have to be excluded.
The plasticity of sea bass ontogeny is general,
and it is not exclusive for sex determination. As
in many other species (Policansky, ’82; Seikai et
al., ’86; Chambers and Leggett, ’87; Polo et al.,
’91; Fuiman et al., ’98), temperature differentially
affects the growth and developmental rate of sea
bass, thus leading to differentiation at smaller TL
under high than under low water temperatures:
Hatching, feeding onset, notochord flexion, fin de-
velopment, and the transitions of the allometric
growth of the body occur at significantly smaller
TL at 20°C than at 13°C or 15°C (Koumoundouros
et al., 2001). Similarly, gonadal differentiation in
different fish species (Conover and Fleisher, ’86;
Strüssmann et al., ’96b, ’97) occurs at significantly
smaller TL under high than under low tempera-
tures. Additionally, male sea bass are not only the
dominant sex at elevated temperatures (Pavlidis
et al., 2000; present study), but they also mature
at a smaller body size than females (Stequert, ’72;
Bruslé and Roblin, ’84; Blàzquez et al., ’95; Kara,
’97). This global character of temperature-induced
ontogenetic plasticity indicates a common princi-
pal or secondary responsible mechanism, with di-
rect consequences for larval survival (discussed
by Koumoundouros et al., 2001) and species re-
production. This mechanism could be the differ-
ential effect of temperature on the developmental
rate of the different cells, tissues, and organs,
which could lead to significant differences between
their interactions during ontogeny.
TSD expression in sea bass is of high interest
not only for aquaculture (i.e., environmental sex
manipulation), but also for natural populations,
given the fact that, although the thermal range
of species reproduction in the natural environment
is 11–15°C (Barnabe, ’76; Mendez et al., ’95;
Mananos et al., ’97), the thermal range of its on-
togeny is 8–20°C (Marangos et al., ’86; Jennings
and Pawson, ’92), thus including the identified
TSD range (Pavlidis et al., 2000; present study).
ACKNOWLEDGMENTS
The authors express their sincere thanks to D.G.
Sfiakianakis and V. Szisch for valuable efforts in
fish sampling. Part of this study was financially
supported by the Commission of the European
Communities, Agriculture and Fisheries (FAIR)
specific RTD program, CT961941, “Early control
for fish production with special reference to muscle
development, gene expression and temperature.”
It does not necessarily reflect its views and in no
way anticipates the Commission’s future policy in
this area.
LITERATURE CITED
Baras E, Prignon C, Gohoungo G, Mélard C. 2000. Pheno-
typic sex differentiation of blue tilapia under constant and
fluctuating thermal regimes and its adaptive and evolution-
ary implications. J Fish Biol 57:210–223.
Baras E, Jacobs B, Mélard C. 2001. Effect of water tempera-
ture on survival, growth and phenotypic sex of mixed
(XX¯XY) progenies of Nile tilapia Oreochromis niloticus.
Aquaculture 192:187–199.
Barnabe G. 1976. Contribution a la connaissance de la biologie
du loup Dicentrarchus labrax (Poisson Serranide) de la re-
gion de Sete. Thèse d’Etat, Université des Sciences et Tech-
niques de Languedoc, Montpellier.
Baroiller JF, Chourrout D, Fostier A, Jalabert B. 1995. Tem-
perature and sex chromosomes govern sex ratios of the
mouthbrooding cichlid fish Oreochromis niloticus. J Exp Zool
273:216–223.
Baroiller JF, Guiguen Y, Fostier A. 1999. Endocrine and en-
vironmental aspects of sex differentiation in fish. Cell Mol
Life Sci 55:910–931.
Beardmore JA, Mair GC, Lewis RI. 2001. Monosex male pro-
duction in finfish as exemplified by tilapia: applications,
problems, and prospects. Aquaculture 197:283–301.
Blàzquez M, Piferrer F, Zanuy S, Carrillo M, Donaldson EM.
1995. Development of sex control techniques for European
sea bass (Dicentrarchus labrax L.) aquaculture: effects of
dietary 17-methyltestosterone prior to sex differentiation.
Aquaculture 135:329–342.
Blàzquez M, Zanuy S, Carillo M, Piferrer F. 1998. Effects of
rearing temperature on sex differentiation in the Euro-
pean sea bass (Dicentrarchus labrax L.). J Exp Zool 281:
207–216.
Blàzquez M, Carrillo M, Zanuy S, Piferrer F. 1999. Sex ra-
tios in offspring of sex-reversed sea bass and the relation-
ship between growth and phenotypic sex differentiation. J
Fish Biol 55:916–930.
Bruslé J, Roblin C. 1984. Sexualité du loup Dicentrarchus
labrax en condition d’elevage controlé. In: Barnabé G,
Billard R, editors. L’Aquaculture du Bar et des Sparidés.
Paris: INRA Publ. p 33–43.
Cataudella S, Civitelli MV, Campanna E. 1973. The chromo-
somes of some mediterranean teleosts: Scorpaenidae,
Serranidae, Labridae, Blenniidae, Gobiidae (pisces-Scorp-
aeniformes, Perciformes). Boll Zool 40:385–389.
Chambers RC, Leggett WC. 1987. Size and age at metamor-
phosis in marine fishes: an analysis of laboratory-reared
winter flounder (Pleuronectes americanus) with a review of
variation in other species. Can J Fish Aquat Sci 44:1936–
1947.
Chatain B, Corrao D. 1992. A sorting method for eliminating
fish larvae without functional swimbladders. Aquaculture
107:81–88.
Conover DO, Fleisher MH. 1986. Temperature sensitive pe-
 ONTOGENETIC SENSITIVITY OF TSD IN SEA BASS
riod of sex determination in the Atlantic silverside, Menidia
menidia. Can J Fish Aquat Sci 43:514–520.
Conover DO, Heins SW. 1987a. The environmental and ge-
netic components of sex ratio in Menidia menidia (Pisces:
Atherinidae). Copeia 1987:732–743.
Conover DO, Heins SW. 1987b. Adaptive variation in envi-
ronmental and genetic sex determination in a fish. Nature
326:496–498.
Conover DO, Kynard BE. 1981. Environmental sex determi-
nation: interaction of temperature and genotype in a fish.
Science 213:577–579.
Desprez D, Mélard C. 1998. Effect of ambient water tem-
perature on sex determinism in the blue tilapia, Oreo-
chromis aureus. Aquaculture 162:79–84.
Francis RC. 1992. Sexual lability in teleosts: developmental
factors. Q Rev Biol 67:1–18.
Fuiman LA, Polling KR, Higgs DM. 1998. Quantifying devel-
opmental progress for comparative studies of larval fishes.
Copeia 1998:602–611.
Janzen FJ, Paukstis GL. 1991. A preliminary test of the adap-
tive significance of environmental sex determination in rep-
tiles. Evolution 45:435–440.
Jennings S, Pawson MG. 1992. The origin and recruitment
of bass, Dicentrarchus labrax, larvae to nursery areas. J
Mar Biol Assoc UK 72:199–212.
Kara MH. 1997. Sexual cycle and fecundity of seabass,
Dicentrarchus labrax, of Annaba Gulf. Cahiers Biol Marin
38:161–168 (cited in Pawson et al., 2000).
Koumoundouros G, Divanach P, Anezaki L, Kentouri M. 2001.
Temperature-induced ontogenetic plasticity in sea bass
(Dicentrarchus labrax). Mar Biol 139:817–830.
Lagomarsino IV, Conover DO. 1993. Variation in environmen-
tal and genotypic sex-determining mechanisms across a lati-
tudinal gradient in the fish, Menidia menidia. Evolution
47:487–494.
Mananos EL, Zanuy S, Carrillo M. 1997. Photoperiodic ma-
nipulations of the reproductive cycle of sea bass (Dicen-
trarchus labrax) and their effects on gonadal development,
and plasma 17β-estradiol and vitellogenin levels. Fish
Physiol Biochem 16:211–222.
Marangos C, Yagi H, Ceccaldi HJ. 1986. The role of tem-
perature and salinity on hatching rate and morphogen-
esis during embryo development in Dicentrarchus labrax
(Linnaeus, 1758) (Pisces, Teleostei, Serranidae). Aquacul-
ture 54:287–300.
Mendez E, Anastasiadis P, Kentouri M, Pavlidis M, Divanach
P. 1995. Preliminary data on spawning activity of five Medi-
terranean teleost species kept in captivity, in Crete (Greece).
In: Castello I, Orvay F, Calderer I, Reig A, editors. Pro-
ceedings of the 5th national congress on aquaculture. Uni-
versity of Barcelona, Barcelona. p 398–403.
Middaugh DP, Hemmer MJ. 1987. Influence of environ-
mental temperature on sex-ratios in the tidewater silver-
side, Menidia peninsulae (Pisces: Atherinidae). Copeia 1987:
958–964.
Pavlidis M, Koumoundouros G, Sterioti A, Somarakis S,
Divanach P, Kentouri M. 2000. Evidence of temperature-
579
dependent sex determination in the European sea bass
(Dicentrarchus labrax L.). J Exp Zool 287:225–232.
Pawson MG, Pickett GD, Witthames PR. 2000. The influence
of temperature on the onset of first maturity in sea bass. J
Fish Biol 56:316–327.
Pieau C, Girondot M, Richard-Mercier N, Dorizzi M, Desvages
G, Zaborski P. 1995. Temperature variation and sex deter-
mination in reptilia. Exp Medicine 13:516–523.
Policansky D. 1982. Influence of age, size, and temperature
on metamorphosis in the starry flounder, Platichthys
stellatus. Can J Fish Aquat Sci 39:514–517.
Polo A, Yufera M, Pascual E. 1991. Effects of temperature on
egg and larval development of Sparus aurata L. Aquacul-
ture 92:367–375.
Roblin C, Bruslé J. 1983. Ontogenèse gonadique et différ-
enciation sexuelle du loup Dicentrarchus labrax, en condi-
tions d’elevage. Reprod Nutr Dev 23:115–127.
Römer U, Beisenherz W. 1996. Environmental determination
of sex in Apistogramma (Cichlidae) and two other freshwa-
ter fishes (Teleostei). J Fish Biol 48:714–725.
Seikai T, Tanangonan JB, Tanaka M. 1986. Temperature in-
fluence on larval growth and metamorphosis of the Japa-
nese flounder Paralichthys olivaceus in the laboratory.
Nippon Suisan Gakkaishi 52:977–982.
Sokal RR, Rohlf FJ. 1981. Biometry. The principles and prac-
tice of statistics in biological research, edition 2. New York:
W.H. Freeman. 857 p.
Sola L, Bressanello S, Rossi AR, Iaselli V, Crosetti D,
Cataudella S. 1993. A karyotype analysis of the genus
Dicentrarchus labrax by different staining techniques. J
Fish Biol 43:329–337.
Stequert B. 1972. Contribution à l’étude du bar Dicentrarchus
labrax (L.) des réservoirs à poissons de la région d’Arcachon.
Thèse de 3ème cycle Université Bordeaux (cited in Pawson
et al., 2000).
Strüssmann CA, Moriyama S, Hanke EF, Calsina Cota JC,
Takashima F. 1996a. Evidence of thermolabile sex determi-
nation in pejerrey. J Fish Biol 48:643–651.
Strüssmann CA, Calsina Cota JC, Phonlor G, Higuchi H,
Takashima F. 1996b. Temperature effects on sex differen-
tiation of two South American atherinids, Odontesthes
argentinensis and Patagonia hatcheri. Env Biol Fishes
47:143–154.
Strüssmann CA, Saito T, Usui M, Yamada H, Takashima F.
1997. Thermal thresholds and critical period of thermo-
labile sex determination in two Atherinid fishes, Odon-
testhes bonariensis and Patagonia hatcheri. J Exp Zool
278:167–177.
Sullivan JA, Schultz RJ. 1986. Genetic and environmental
basis of variable sex ratios in laboratory strains of Poe-
ciliopsis lucida. Evolution 40:152–158.
Wang L-H, Tsai C-L. 2000. Effects of temperature on the de-
formity and sex differentiation of tilapia, Oreochromis
mossammbicus. J Exp Zool 286:534–537.
Yamamoto E. 1999. Studies on sex-manipulation and produc-
tion of cloned populations in hirame, Paralichthys olivaceus
(Temminck et Schlegel). Aquaculture 173:235–246.