Embryonic and larval development of black skirt tetra

(Gymnocorymbus ternetzi, Boulenger, 1895) under

laboratory conditions
I

hsan C elik
1
, Pnar C elik
1
, S u kran Cirik
1
, Mert Gu rkan
2
& Sibel Hayretdag
2
1
Department of Aquaculture, Fisheries Faculty, C anakkale Onsekiz Mart University, C anakkale, Turkey
2
Department of Biology, Faculty of Science, C anakkale Onsekiz Mart University, C anakkale, Turkey
Correspondence: I

C elik, Department of Aquaculture, Fisheries Faculty, C anakkale Onsekiz Mart University, Terzioglu Campus,
17100 C anakkale, Turkey. E-mail: celik_ihsan@yahoo.com
Abstract
The embryonic and larval development of black
skirt tetra, Gymnocorymbus ternetzi, are described
under controlled laboratory conditions. In addition,
major histomorphological changes and the allomet-
ric growth patterns during larval development have
been described. The laboratory-reared broodstock,
that is 1 year of age, were spawned. Hatching
occurred 2021 h after spawning at 24 0.5C.
The cleavage was nished in 2 h and the early blas-
tula stage occurred at 2:04 hours after spawning.
The gastrulation started at 3:20 hours and 30%
epiboly was observed at 3:34 hours after spawning.
Eight-somite stage was observed at 08:33 hours.
And embryonic developmental stage was completed
at 21 h after spawning. The newly hatched larvae
were 1442 14.3 lm in mean total length (TL).
The mouth opened at 3 days after hatching (DAH).
The yolk sac had been totally absorbed and the
larvae started to swim actively within 34 days.
Notochord exion began at 11 DAH. The metamor-
phosis was completed and the larvae transformed
into juveniles at 32 DAH. In this paper, the full
developmental sequence from egg to juvenile of
G. ternetzi is described for the rst time.
Keywords: Gymnocorymbus ternetzi, embryonic
development, larval development, morphological
characteristics, allometric growth
Introduction
Characidae is a family of freshwater subtropical
and tropical sh, found in southwestern Texas,
Mexico, Central and South America and it is a
large family that comprises about 152 genera and
776 species (Nelson 1994). The black skirt tetra
(Gymnocorymbus ternetzi) is just one of many sh
in the group of tetras, Characins, is traded in the
ornamental sh industry. It is a popular species in
the trade of freshwater ornamental sh, since it is
attractive in appearance, undemanding in mainte-
nance and easily bred (Frankel 2004; Uma &
Chandran 2008). Information on embryonic and
larval development of sh is a fundamental key
which enables a closer approach to their biology
and taxonomy (Reynalte-Tataje, Zaniboni-Filho &
Esquivel 2004). Morphological features are very
important as they furnish information of life his-
tory of sh and they provide critical parameters to
hatchery production (Martinez & Bolker 2003;
Silva 2004). Early life history characters of sh
can be used in assessing phylogenetic relationships
(Richards & Leis 1984; Stiassny & Mezey 1993;
Britz 1997; Meijide & Guerrero 2000)). In addi-
tion, studies on embryonic and larval development
of any sh species can be useful in directing the
husbandry efforts of sh breeder to the specic
state and requirements of each development
stage (Marimuthu & Haniffa 2007). There is vast
literature on embryonic and larval stages of sh,
distributed among the elds of aquaculture,
applied ecology, behavioural ecology, biological
oceanography, comparative functional morphology
and physiology, sheries science, limnology and
systematic ichthyology (Takeshita, Onikura,
Matsui & Kimura 1997; Webb 1999; Arvedlund,
McCormick & Ainsworth 2000; Borges, Faria, Gil,
Goncalves & Almada 2003; Martell, Kieffer &
2011 Blackwell Publishing Ltd 1260
Aquaculture Research, 2012, 43, 12601275 doi: 10.1111/j.1365-2109.2011.02930.x
Trippel 2005; Marimuthu & Haniffa 2007; Du,
Wang, Jiang, Liu, Wang, Li & Zhang 2010). How-
ever, detailed study about the embryonic and lar-
val development of characins is scarce. In
addition, information is lacking concerning ontog-
eny of black skirt tetra from egg to juvenile. In the
present study, the embryonic and larval develop-
ment of laboratory-reared black skirt tetra (G.
ternetzi) from egg to juvenile are described in
detail for the rst time. In addition, major histo-
morphological changes and the allometric growth
patterns during larval development have been
identied.
Materials and methods
Broodstock maintenance
One-year-old black skirt tetras (G. ternetzi) were
used as broodstock in the experiment. They were
fed with commercial ornamental sh feeds (Tetr-
amin Granulat, Tetra, Germany; Protein: 46%, Oil:
12%, Fibre: 3%, Ash: 11%, Moisture: 8%), three
times a day. During broodstock culture, water
temperature, pH and conductivity were monitored
daily at 24 0.5C, between 6.0 and 6.5 and
between 100 and 200 lS respectively. Water
temperature was controlled with additional sub-
merged heaters. The photoperiod was maintained
at 9L/15D by uorescent lighting (lights on;
07:0018:00 hours). Broodstocks were kept in
40 L glass aquariums. Three pairs (3 males/3
females) were randomly selected from broodstock
tank and were placed into a 15 L spawning tank
late in the afternoon. Spawning was observed the
next day around dawn and lasted 13 h. Eggs and
larvae were obtained from three pairs of brood-
stocks.
Observations and measurements of embryos and
larvae
Fertilized eggs were collected immediately after
spawning and maintained at 24 0.5C. Some of
them were transferred into a beaker (500 mL) for
embryonic development observations. The others
were maintained in 15 L aquaria at 24 0.5C.
Eggs were observed from spawning to hatching
under an Olympus BX51 research microscope
(Hatagaya, Shibuya-ki, Tokyo, Japan) and photo-
graphed using a colour video camera (Q Imaging,
Micropublisher 3.3 RTV, Burnaby, BC, Canada).
Embryonic development stages were identied
according to Kimmel, Ballard, Kimmel, Ullman
and Schilling (1995).
Larvae were fed once a day with Artemia sp.
(INVE Aquaculture Inc., Dendermonde, Belgium)
until the end of the experiment at 32 days after
hatching (DAH). They were randomly sampled
(n = 5) daily from hatch to 18 DAH and at 1-day
interval from 18 to 32 DAH. These specimens were
observed under an Olympus SZX7 zoom stereomi-
croscope, photographed by a colour video camera
and measured using image analysis program (Q
Capture Pro, version 5.1.1.14, Dendermonde, Can-
ada). On the other hand, they were used for obser-
vations on general morphology and for the
following morphometric measurements (mm): body
depth (BD), eye diameter (ED), head length (HL),
pre-anal length (PAL), Pre-anal myomer length
(PrAM), post-anal myomer length (PoAM), snout
length (SnL), tail length, total length (TL), trunk
length (Fig. 1) Larval developmental stages were
identied according to Kendall, A.W., Ahlstrom
and Moser (1984) and differentiated into four peri-
ods I: yolk-sac larva, II: preexion larva, III: exion
larva and IV: postexion larva.
Figure 1 Morphometric characters measured in the black skirt tetra larvae.
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C elik et al.
Allometric growth patterns were calculated as a
power function of TL (Fuiman 1983) with the
exponent and intercept obtained from linear regres-
sions on log-transformed data (Gisbert, Merino,
Muguet, Bush, Piedrahita & Conklin 2002). The al-
lometric equation Y = aX
b
of BD, ED, HL, PAL,
PrAM, PoAM, SnL, tail length and trunk length on
TL was estimated. Here Y is the dependent variable
(measured character), X is the independent vari-
able (TL), a is the intercept and b is the growth
coefcient. When isometric growth occurred,
b = 1, a positive allometric growth occurred when
b > 1 and a negative one when b < 1.
Histological observations
For histological evaluations, larvae were randomly
collected (n = 10) daily from hatch (0 DAH) to 10
and every 2 days from 10 to 32 DAH. These speci-
mens were xed in Bouins solution and 70% alco-
hol, dehydrated through a series of alcohol
concentrations, cleared in xylene and embedded in
(a) (b) (c)
(d) (e) (f)
(g) (h) (i)
(j)
(k) (l)
(m) (n)
(o)
(p) (r) (s)
Figure 2 The stages of embryonic development Gymnocorymbus ternetzi: (a) 2-blastomere stage; (b) 4-blastomere
stage; (c) 8-blastomere stage; (d) 16-blastomere stage; (e) 32-blastomere stage; (f) early blastula stage; (g) late blas-
tula stage; (h) early gastrula stage; (i) 30% epiboly; (j) 50% epiboly; (k) 75% epiboly; (l) 8-somite stage; (m) 11-
somite stage; (n) 13-somite stage; (o) otic capsule; (p) muscular effect; (q) otolith appearance; (r) hatching, 3 h after
hatching. Developmental stages were determined by comparison with standard zebrash embryonic stages as
described by Kimmel et al. (1995). Scale bars, ar: 500 micron, s: 1mm.
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1262
Embryonic and larval development of black skirt tetra I

C elik et al. Aquaculture Research, 2012, 43, 12601275

parafn wax. Wax blocks were cut using a micro-
tome (Slee, Cut5062, Germany) at 5lm. Sagittal
sections were stained with Gills haematoxylin/
eosin (HE) procedures for general histology. Sec-
tions were observed under a light microscope
(Olympus BX50) to describe the larval development
and photographed using a colour video camera.
Results
Embryonic development
The eggs were adhesive, demersal and spherical in
shape. The eggs ranged in diameter from 930.23
to 1063.95 lm, with a mean of 977.36
45.86 lm (n = 19). The egg capsule was transpar-
ent, while the yolk was brownish. The cleavage
of eggs was meroblastic and the rst cleavage
(two-celled stage) occurred within 0:30 hours after
spawning (Fig. 2a). Blastodisc divided to form two
equal cells. The second cleavage occurred 0:43
hours and four blastomeres are clearly observed
(Fig. 2b). Blastodisc divided via meridional cleav-
age to form four equal cells The third and fourth
cleavages dene 8 and 16 respectively (Fig. 2c
and d). The eggs cleaved into 8 and 16 cells
respectively. The third cleavage is horizontal and
results in a 2 9 4 array (Fig. 2c). The forth cleav-
ages occur in two separate planes, cleavage furrow
parallel to second cleavage plane and results in a
4 9 4 array (Fig. 2d). Eight and 16 cell stages
were observed at 0:50 hours and 1:04 hours
respectively (Fig. 2c and d). The fth cleavage
took place after 1:10 hours from spawning
(Fig. 2e). Blastoderm divided via meridional cleav-
age into 32 cells and the 32 blastomeres are
already formed. The cells became smaller and were
arranged irregularly. The early blastula stage
occurred at the vegetal pole 2:04 hours after
spawning (Fig. 2f). At this stage, the crowded cells
expanded over the yolk and the blastomeres were
divided asynchronously. The late blastula stage
consists of a multicellular blastomere (Fig. 2g) and
fully completed at approximately 2:303:00 hours.
The gastrulation started at 3:20 hours after
spawning (Fig. 2h). Blastoderm cells spread over
the yolk and epibolic cells increased at this stage.
The embryo reached 30% epiboly at 3:34 hours
after spawning and the blastoderm covered 30% of
the yolk (Fig. 2i). 50% and 75% epiboly stages
were completed at 4:10 hours and 5:30 hours
respectively (Fig. 2j and k). The segmentation
stage was characterized by the sequential forma-
tion of the somites and lasted till just prior to
hatching. Eight-somite stage in black skirt tetra
was observed in the central part of the embryo at
08:33 hours (Fig. 2l). Eleven pairs of somites
formed at 09:00 hours and the number of somites
from stage 11 to stage 13 increased within 1 h
(Fig. 2m and n). The formation of the otic capsule
Table 1 Embryonic development stages of Gymnocorymbus ternetzi at 24 0.5C
Main stages Substages Time (h:min) Description Figure
Zygote 2 cells 0:30 First cleavage, blastodisc divided via meridional
cleavage to form two equal cells
1a
4 cells 0:43 Second cleavage, dividing the blastodisc into 4 blastomeres 1b
8 cells 0:50 Third cleavage 1c
16 cells 1:04 Fourth cleavage, 16 blastomeres can be seen 1d
32 cells 1:10 Fifth cleavage 1e
Blastula Early Blastula 2:04 Blastomeres continued to divide but they were less
synchronously
1f
Late Blastula 2:26 Epibolic cells increase 1g
Gastrula Early Gastrula 3:20 Blastoderm cells begin to spread over the yolk 1h
%30 Epiboly 3:34 Germ ring epiboled 1/3 of yolk sac 1i
%50 Epiboly 4:10 Germ ring epiboled 1/2 of yolk sac 1j
%75 Epiboly 5:30 75% coverage of the yolk cell by the blastoderm 1k
Segmentation 8 Somite 8:33 1l
11 Somite 9:00 1m
13 Somite 10:01 1n
Pharyngula Otic capsule 13:15 Otic capsule formed 1o
Muscular effect 15:30 Embryo begins to spin 1p
Pre-hatching stage 18:35 The embryo shows conspicuous muscular contractions 1q
Hatching 3 h after hatching 24:00 Pre-larva is 3 h after hatching 1r
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C elik et al.
started at 13:15 hours and embryo began to spin
at 15:30 hours after spawning (Fig. 2o and p).
The eye development and brain differentiation
(forebrain, midbrain and hindbrain) has taken
place (Fig. 2q). Hatching rates were 8590% in
aquarium at 20 h after spawning. The embryonic
development completed at 21 h (Table 1).
(a)
(b)
(c)
(d)
(e)
(f)
Figure 3 Larval development of Gymnocorymbus terne-
tzi. (a) Post-hatching stage (7 h); (b) yolk-sac stage (1
DAH); (c) yolk-sac stage, the gas bladder was formed but
not completely lled (2 DAH); (d) opened-mouth stage
(3 DAH); (e) preexion larva, exogenous feeding (5
DAH); (f) preexion larva (7 DAH). Scale bars = 1 mm.
(a)
(b)
(c)
(d)
(e)
(f)
Figure 4 Larval development of Gymnocorymbus terne-
tzi. (a) Flexion stage, Notochord exion started (11
DAH); (b) exion stage, the notochord was completely
exed (12 DAH); (c) postexion larva, swim bladder
with two chambers was visible (17 DAH); (d) postex-
ion larva (19 DAH); (e) postexion larva (26 DAH); (f)
end of metamorphosis (30 DAH). Scale bars (g. a, b,
c, d, e) = 1 mm; Scale bar (g. f) = 500lm.
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C elik et al. Aquaculture Research, 2012, 43, 12601275

Larval Development
Morphological observations
Newly hatched larvae
Newly hatched larvae in the post-hatching stage
were laterally compressed and initially elongated.
The TL of newly hatched larvae was
1442 14.3 lm. The mouth was closed and the
yolk sac was 30% of the total body length
(Fig. 3a). Two otoliths which were like black dots
within the otic vesicles were visible (Fig. 3a). The
body of the newly hatched larvae was transparent
but pigmentation has started in the posterior part
of the yolk sac (Fig. 3a). The eyes were still unpig-
mented.
1DAH (TL: 3.03 0.02 mm)
The mouth and anus were closed and the undiffer-
entiated alimentary tract appeared as a straight
tube (Fig. 3b). Eyes were not pigmented and sev-
eral stellate melanophores were scattered over the
surface of body and the yolk sac. The primordial
n fold was well developed in the sagittal plane
but no ns were differentiated (Fig. 3b). The yolk
sac was ovoid and 20% the total body length
(Fig. 3b).
2 DAH (TL: 3.50 0.09 mm)
The yolk sac has become smaller (12% the TL)
(Fig. 3c). Initial ination was observed; the oval-
shaped swim bladder was rst seen in 2 DAH. Pig-
mentation has increased over the eyes and the
body but they are still translucent. Black melano-
phores were scattered on the head region, ventral
and dorsal side of the body (Fig. 3c). The urinary
bladder is visible and seen near the anus. The pri-
mordial n was slightly differentiated but, no anal
and dorsal ns were differentiated but pectoral n
bud was present (Fig. 3c). The larvae could not
swim actively but short periods of swimming were
observed.
34 DAH (TL: 3.64 0.124.03 0.06 mm)
At 3 DAH, the mouth and anus opened (Fig. 3d).
The yolk sac has been completely absorbed and
Figure 5 The main events of larval development in black skirt tetra (Gymnocorymbus ternetzi).
y = 3.0053e
0.0531x
R
2
= 0.9694, n = 112
0
5
10
15
20
25
30
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34
Age (DAH)
T
o
t
a
l

l
e
n
g
t
h

(
m
m
)
Yolk sac Preflexion Postflexion
Flexion
Juvenile
Figure 6 Growth of black skirt tetra larvae from hatch to 32 DAH. Each point represents the mean total
length SD.
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Aquaculture Research, 2012, 43, 12601275 Embryonic and larval development of black skirt tetra I

C elik et al.
the larvae started to swim actively at 34 days.
The larvae started to feed exogenously within
3 days at 24 1C. The eyes were pigmented.
The larvae have a one-chambered swim bladder.
The notochord end is not exed (Fig. 3d).
57 DAH (TL: 4.29 0.074.37 0.05 mm).
The eyes became very prominent and were fully
pigmented (Fig. 3e). Second ination of swim
bladder did not occur. It formed as a single cham-
ber, increased in size and extended posteriorly. The
larvae have still primordial ns (Fig. 3e and f).
The notochord end was not exed (Fig. 3e and f).
The larvae could swim very well. Pigmentation
has increased on the head and lateral parts of the
body, black pigments were dominant, but yellow
pigments were also present (Fig. 3f).
1112 DAH (TL: 4.97 0.155.78 0.23 mm).
Primordial n was still present (Fig. 4a). Pectoral
ns were well developed. Dorsal and anal ns have
begun early differentiation (Fig. 4a and b). The
caudal-n rays begin to form. At 11 DAH, the
notochord end was slightly exed but exion is
more obvious at 12 DAH. Swim bladder increased
in size and extended posteriorly (Fig. 4b). The lar-
vae could swim very well. There were clusters of
pigment over the body but pigmentation was more
concentrated on head region (Fig. 4b).
1517 DAH (TL: 5.75 0.166.09 0.27 mm).
Second ination of swim bladder occurred between
15 DAH and 17 DAH. Swim bladder with two
chambers completely lled (Fig. 4c). Anal and dor-
sal ns begin to develop but have no rays but cau-
dal n rays were more developed (Fig. 4c). Body
shape and pigmentation pattern were not similar
to the adult sh. The stomach of larvae contained
food, ventral region of larvae was swollen and
orange.
1926 DAH (TL: 7.83 0.8211.67 1.37 mm).
At 1922 DAH, dorsal and anal ns have differen-
tiated (Fig. 4d and e). Adipose n began to form
between the dorsal and caudal ns at 22 and 23
DAH. Pigmentation has increased over the body
but the ventral trunk region was still translucent
and food particles (Artemia) could be seen in the
digestive tract. The adipose n was more obvious
at 25 and 26 DAH (Fig. 4e). Dorsal and anal ns
were more developed with the separated rays and
caudal n was forked (Fig. 4e). Body depth was
more than triple that of the previous stage and the
body shape of larvae has approached an adult
shape.
3032 DAH (TL: 14.67 0.9220.40 1.30 mm).
The body shape of larvae and pigmentation pat-
tern were similar to those of the adult (Fig. 4f).
The body was almost completely covered with pig-
ment (Fig. 4f). Larvae have a dark grey to silver
body with black vertical bars but black and grey
pigments were dominant. Two characteristic black
vertical bars were visible on posterior side of the
gills (Fig. 4f). Morphological metamorphosis was
completed and the larvae had completely trans-
formed into juveniles. The main events during
larval development of black skirt tetra are summa-
rized in Fig. 5.
Figure 7 Growth coefcients of head, trunk and tail
length during larval development stage. Each graph
represents the growth coefcients during a total length
(TL) interval.
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C elik et al. Aquaculture Research, 2012, 43, 12601275

y = 0.3783x
1.1261
R
2
= 0.9807, n = 107
0
2
4
6
8
10
12
14
16
0.00 5.00 10.00 15.00 20.00 25.00 0.00 5.00 10.00 15.00 20.00 25.00
0.00 5.00 10.00 15.00 20.00 25.00 0.00 5.00 10.00 15.00 20.00 25.00
0.00 5.00 10.00 15.00 20.00 25.00
Total length (mm)
T
a
i
l

l
e
n
g
t
h

(
m
m
)
y = 0.6101x
0.9197
R
2
= 0.9808, n = 107
0
2
4
6
8
10
12
Total length (mm)
P
A
L

(
m
m
)
y = 0.3635x
0.9329
R
2

= 0.9611, n = 107
0
2
4
6
8
10
Total length (mm)
P
r
A
M

(
m
m
)
y = 0.0296x
1.2008
R
2
= 0.8903, n = 112
0
0.2
0.4
0.6
0.8
1
1.2
1.4
Total length (mm)
S
n
L

(
m
m
)
y = 0.4388x
0.9477
R
2
= 0.9732, n = 107
0
2
4
6
8
10
12
Total length (mm)
P
o
A
M

(
m
m
)
Figure 9 Allometric growth equations and relationship between ve body segments and total length in black skirt
tetra during larval development period (from hatch to 32 DAH). PAL, Pre-anal length; PrAM, Pre-anal myomer
length; PoAM, Post-anal myomer length; SnL, snout length.
y = 0.4333x
0.8191
R
2
= 0.9391, n = 107
0
1
2
3
4
5
6
Total length (mm)
T
r
u
n
k

l
e
n
g
t
h

(
m
m
)
y = 0.1644x
1.1588
R
2
= 0.9525, n = 112
0
1
2
3
4
5
6
7
0.00 5.00 10.00 15.00 20.00 25.00
0.00 5.00 10.00 15.00 20.00 25.00 0.00 5.00 10.00 15.00 20.00 25.00
0.00 5.00 10.00 15.00 20.00 25.00
Total length (mm)
H
e
a
d

l
e
n
g
t
h

(
m
m
)
y = 0.0639x
1.1215
R
2
= 0.9737, n = 112
0
0.5
1
1.5
2
2.5
Total length (mm)
E
y
e

d
i
a
m
e
t
e
r

(
m
m
)
y = 0.1124x
1.3751
R
2
= 0.9584, n = 112
0
2
4
6
8
10
Total length (mm)
B
o
d
y

d
e
p
t
h

(
m
m
)
Figure 8 Allometric growth equations and relationship between four body segments and total length in black skirt
tetra during larval development period (from hatch to 32 DAH).
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1267
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C elik et al.
Allometric growth
Growth of the black skirt tetra larvae followed an
exponential curve during the larval stages and is
represented by the equation y = 1.79e
0.051x
(R
2
= 0.97, n = 112) where y is total length (TL)
mm and x is DAH (Fig. 6). Four larval development
stages were observed after hatching; yolk-sac
larvae, preexion larvae, exion larvae and postex-
ion larvae. The yolk sac has been completely con-
sumed at 4 DAH, when TL was 4.03 0.14 mm.
Notochord has been exed between 10 DAH and
12 DAH, at 5.34 0.57 mm. All the meristic char-
acters were completely developed and juvenile stage
(a)
(b)
(c)
Figure 10 Sagittal sections of black skirt tetra larvae.
(a) 2 DAH (Olympus BX51 1009), (b) 3 DAH (Olym-
pus BX51 409), (c) 4 DAH (Olympus BX51 409). at,
alimentary tract; e, eye; ga, gill arches; l, liver; n, noto-
chord; oe, oesophagus; ph, pharynx; s, stomach; sb,
swim bladder; t, teeth; ys, yolksac.
(a)
(b)
(c)
Figure 11 Sagittal sections of black skirt tetra larvae.
(a) 16 DAH (Olympus SZX7 zoom stereo microscope
209), (b) 24 DAH (Olympus SZX7 zoom stereo micro-
scope 12.5x), (c) 32 DAH (Olympus SZX7 zoom stereo
microscope 89). df, dorsal n rays; e, eye; g, gill; ga,
gill arches; gl, gill lamellae; i, intestine; l, liver; m,
mouth; n, notochord; oe, oesophagus; ph, pharynx; s,
stomach; sb, swim bladder; sb1, rst chamber of swim
bladder; sb2, second chamber of swim bladder; t, teeth.
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1268
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C elik et al. Aquaculture Research, 2012, 43, 12601275

started at 32 DAH, TL was 20.40 2.25 mm at
32 DAH.
In the yolk-sac stage, the head, trunk and tail
length had negative allometric growth in relation
to TL (b = 0.41, b=0.02, b = 0.25 respectively)
(Fig. 7). During the preexion stage, they still
showed negative allometric growth (b = 0.45,
b = 0.39, b = 0.15 respectively) (Fig. 7). The head,
trunk and tail length were positively allometric in
the exion stage (b = 2.43, b = 3.29, b = 2.21
respectively) (Fig. 7). In the postexion stage,
growth of trunk length was negatively allometric
(b = 0.87), while growth in the head and tail
length were positively allometric (b = 1.16,
b = 1.5 respectively) (Fig. 7).
Allometric growth equations between nine mea-
sured body segments and total length during lar-
val development stage (032 DAH) are presented
(Figs 8 and 9). Growth of trunk in length was
negatively allometric from hatch to 32 DAH
(a = 0.43, b = 0.82, R
2
= 0.94, n = 107), while
the growth coefcients of HL, ED and BD were pos-
itively allometric (a = 0.16, b = 1.16, R
2
= 0.95,
n = 112; a = 0.06, b = 1.12, R
2
= 0.97, n = 112
and a = 0.11, b = 1.38, R
2
= 0.96, n = 112
respectively) (Fig. 8).
The characters PAL and PrAM showed negative
allometry (a = 0.61, b = 0.92, R
2
= 0.98, n = 107
and a = 0.36, b = 0.93, R
2
= 0.96, n = 107
respectively), while tail length and SnL exhibited
positive allometric growth (a = 0.38, b = 1.13,
R
2
= 0.98, n = 107 and a = 0.03, b = 1.20,
R
2
= 0.98, n = 112 respectively) (Fig. 9). PoAM
showed isometric allometric growth (a = 0.44,
b = 0.95, R
2
= 0.97, n = 107).
Histological observations
At 2 DAH; the mouth was closed, the alimentary
tract was distinct as a straight tube. The yolk sac
has become smaller (Fig. 10a). The swim bladder
was formed and initial ination was observed at 2
DAH (Fig. 10a). At 3 DAH; the yolk was not
totally consumed. The mouth and anus were open
(Fig. 10b). Gill lamellae were observed in laments
carried by gill arches (Fig. 10b). The swim bladder
increased in size and extended posteriorly. Larvae
began to feed exogenously before complete resorp-
tion of the yolk sac (Fig. 10b). 4 DAH; yolk has
been completely consumed (Fig. 10c). The larvae
were capable of feeding on Artemia and there was
presence of food material in the stomach. The
swim bladder with one chamber increased in size.
Eye pigment was concentrated and opaque. The
liver could be seen on the ventral side of the swim
bladder (Fig. 10c). Four pairs of gill arches with
laments were obvious (Fig. 10c). The swim blad-
der increased in size and extended posteriorly and
second ination of swim bladder in larvae
occurred between 15 and 18 DAH (Fig. 11a). No
histological difference was observed between the
anterior intestine and the posterior intestine
(Fig. 11a). 24 DAH; the dorsal and anal n rays
formed. The swim bladder with two chambers
could be seen (Fig. 11b). 3032 DAH; Metamor-
phosis was completed and the larvae completely
transformed into juveniles. Large sized food parti-
cles were observed in the stomach and intestine
(Fig. 11c). The body depth increased.
Discussion
In this study, the embryonic and larval develop-
ment of the laboratory-reared black skirt tetra (G.
ternetzi) are described. For the rst time, the full
developmental sequence from egg to juvenile in
controlled aquarium conditions is also stated. In
addition, allometric growth of some body parts
was studied. Embryonic development lasted 21 h
and larval development lasted about 3032 days
at 24 0.5C.
Egg size is an important consideration for egg
and larval quality during incubation and rearing
in aquaculture. The average diameter of most
ornamental sh eggs are around 0.8 mm, how-
ever, the size range is wide (Watson & Chapman
2002). Egg diameters of some aquarium sh were
reported as; 0.4 mm for gobies (Watson & Chap-
man 2002), 0.740.90 mm for Hyphessobrycon ser-
pae (Characidae) (Cole & Haring 1999), 0.9 mm
for Microgeophagus ramirezi (Cichlidae) (Coleman &
Galvani 1998), 1.1 mm for Symphysodon discus
(Cichlidae) (Coleman & Galvani 1998), 1.01.6 mm
for Symphysodon spp. (C elik 2008), 4.0 mm for
Tropheus moori (Cichlidae) (Coleman & Galvani
1998), 2.9 mm for Cyathopharynx fucifer (Cichli-
dae) (Coleman & Galvani 1998), 1.65 0.5 for
Cichlasoma dimerus (Cichlidae) (Meijide & Guerrero
2000), 1.18 0.05 mm for Capoeta tetrazona (Cyp-
rinidae) (Tamaru, Cole, Bailey & Brown 2001),
1.41.6 mm for Carassius auratus (Cyprinidae)
(Savas, Sener & Yldz 2006), 0.75 mm for Puntius
conchonius (Cyprinidae) (Bhattacharya, Zhang &
Wang 2005), 0.7 mm for Danio rerio (Cyprinidae)
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1269
Aquaculture Research, 2012, 43, 12601275 Embryonic and larval development of black skirt tetra I

C elik et al.
(Kimmel et al. 1995), 1.47 0.20 mm for Corydo-
ras aeneus (Callichthyidae) (Huysentruyt & Adria-
ens 2005), 1.799 0.0214 mm for Corydoras
paleatus (Callichthyidae) (U

nal & Aral 2006), 1.5

2.0 mm for some ornamental catsh (Watson &
Chapman 2002) and from 0.89 0.97 mm to
1.03 0.06 mm for Trichogaster pectoralis (Osphro-
nemidae) (Morioka, Ito & Kitamura 2010). The egg
size and fecundity depend on several factors, for
example broodstock age, broodstock size, feed and
water quality. The variety of reproductive strate-
gies of sh can cause great differences among
species in the number of eggs, egg size (Andrade-
Talmelli, Kavamoto, Romagosa & Fenerich-Verani
2001) but the eggs of a sh species are in a com-
mon size range. The eggs of black skirt tetra are
spherical, adhesive, demersal and have approxi-
mately 0.98 0.05 mm average diameter. The
number of eggs per spawn, per female was around
150200 and fertilization rate was about 90%.
In most sh species the blastomeres are regular
in size and shape (Hall 2008). In the black skirt
tetra, rst ve cleavages divided the blastodisc into
32 equal-sized blastomeres at the animal pore and
horizontal cleavage occurred between 64 and 128
cell stages (after the fth divison). In the zebrash
Danio (Brachydanio) rerio (Kimmel et al. 1995), in
the Atlantic cod Gadus morhua (Hall, Smith &
Johnston 2004; and in the Cichlasoma dimerus
(Meijide & Guerrero 2000) the rst horizontal
cleavage occurs at the sixth cleavage, between the
32 and the 64 cell stages. In the medaka Oryzias
latipes, it occurs between the 16 and 32 cell stages
(Iwamatsu 1994). It occurs even earlier in the
Holostean sh Amia calva (between the 8 and the
16 cell stages) (Ballard 1986; Nakatsuji, Kitano,
Akiyama & Nakatsuji 1997). and in the ice goby
Leucopsarion petersii (between the 4 and the 8 cell
stages) (Nakatsuji et al. 1997). Theoretical knowl-
edge of embryonic development stages might be
useful for incubation management with regard to
environmental variables, thus larvae malformation
and low productivity in captivity can be prevented
(Alves & Moura 1992). Furthermore, the studies
on embryonic and early larval development are
important to the successful rearing of larvae for
large-scale seed production and aquaculture (Khan
& Mollah 1998; Koumoundouros, Divanach &
Kentouri 2001; Borcato, Bazzoli & Sato 2004;
Rahman, Rahman, Khan & Hussain 2004). In
addition, it was reported that information on egg
characteristics was important for the tness of lar-
vae (Saillant, Chatain, Fostier, Przybyla & Fauvel
2001).
Early blastula was observed 2:04 hours later.
Teleost gastrulation was morphologically charac-
terized by the presence of a germ ring (Arezo,
Pereiro & Berois 2005). In this study, gastrulation
was observed at 3:20 hours and 30% epiboly
began 3:34 h. G. ternetzi embryo reached the
eight-somite stage at 8:33 hours and reached the
pre-hatching stage at 18 h with the otic capsule,
head, primordial n, tail. All the embryos had
hatched after 2122 h after the fertilization at
24 0.5C. It was reported that small differences
in temperature ( 2C) was important on larval
survival of lemonpeel angelsh Centropyge avissi-
mus (Olivotto, Holt, Carnevali & Holt 2006). This
rule is the same for G. ternetzi larvae and for many
ornamental sh. G. ternetzi can spawn between
23C and 30C, eggs were obtained from adult
broodstock maintained at 24 0.5C in this
study. So, the water temperature was kept con-
stant during larval rearing (24 0.5C). Fish
embryos and larvae are generally more sensitive
to temperature change than older sh (Wood &
Mc Donald 1996). It is known that temperature
affects growth (Hunt von Herbing & Boutilier
1996; Wood & Mc Donald 1996; Mommsen 2001;
Martell et al. 2005), metabolic activity, embryonic
and larval mortality (survival) (Hanel, Karjalainen
& Wieser 1996; Bermudes & Ritar 1999; Martell
et al. 2005), energy utilization (Finn, Rnnestad,
van der Meeren & Fyhn 2002), yolk-sac consump-
tion (Fukuhara 1990), enzyme activity (Papout-
soglou & Lyndon 2005) on embryonic and larval
development in sh. For example, while G. ternetzi
eggs hatch within 2021h after fertilization at
24 0.5C, the incubation time can be shorter at
28C (1617 h). On the other hand, most of the
eggs and larvae died under 22C and above 30C.
Minimum, maximum and optimum temperature
levels for growth and survival of G. ternetzi larvae
must be determined by future studies.
At hatching, the larva was 1.44 mm in total
length. The larva reached a size of 1420 mm
between 30 and 32 days and larval development
had completed. Eggs and larvae are not guarded
by the parents; within the rst 3 days of hatching,
the larvae were inactive but short periods of swim-
ming were observed. They started swimming freely
within 34 days. On the other hand, it was
reported that G. ternetzi larvae hatched 2024 h
after spawning and were free-swimming 4872 h
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1270
Embryonic and larval development of black skirt tetra I

C elik et al. Aquaculture Research, 2012, 43, 12601275

after hatching at 24C (Frankel 2004). While
many marine sh larvae had two kinds of energy
reserves, yolk and oil globule (Bjelland & Skiftesvik
2006), black skirt tetra has only yolk sac. The
yolk sac is depleted within 34 days and the lar-
vae start to feed exogenously before complete
absorption of the yolk sac. Mouth opening was
formed on the third day. Compared to other fresh-
water ornamental sh species (Nandini & Sarma
2000), larvae of G. ternetzi have small mouth and
consume a small size range of prey. So, rotifer and
infusoria must be preferred as rst feed. In this
study, infusoria that is the smallest of live foods
for sh larvae was used for rst food until 7 DAH.
Black skirt tetra larvae can catch and eat Artemia
at 5 DAH, so Artemia nauplii has been used from
5 DAH to 32 DAH.
There are two types of swim bladders, physost-
omous and physoclistous (Moyle & Cech 2000;
Trotter, Pankhurst & Battaglene 2004; Govoni &
Forward 2008). According to Moyle and Cech
(2000), characins that include black skirt tetras,
cyprinids, salmonids, pike, catsh, mormyrids and
eels are sh species with physostomous swim blad-
der. Physostomous swim bladders are connected to
the alimentary tract and physostomous larvae
inate their swim bladder with atmospheric air
(Moyle & Cech 2000; Trotter et al. 2004; Govoni
& Forward 2008; Perlberg, Diamant, Or & Zilberg
2008). In this study, the swim bladder was rst
discernible at 2 DAH. Larvae could not start free-
swimming and anus was closed at that time. Mor-
phological and histological ndings indicated that,
the swim bladder was lled with a little air or
uid. It was reported that larvae inate the swim
bladder with gas from the digestion of organic
material in the gut. Although this has been ruled
out, it is possible (Pelster 2004). In larval culture,
swim bladder ination requires special attention as
it affects feeding activity (O

nal, C elik & Cirik

2010). It was reported that swim bladder ination
coincided with yolk sac depletion and the start of
feeding activity in most species such as discus
(Symphysodon spp.,), Nile tilapia (Oreochromis niloticus),
Mozambique tilapia (Oreochromis mossambicus), Dover
sole, Solea solea and striped trumpeter, Latris line-
ata (Boulhic & Gabaudan 1992; Marty, Hinton &
Summerfelt 1995; Trotter, Pankhurst & Hart
2001; O

nal et al. 2010). In contrast, it was

reported that yolk depletion and the start of feed-
ing activity did not appear to correlate distinctly
with swimbladder ination in some species such as
freshwater angelsh (Pterophyllum scalare) (Zilberg,
Or, Rabinski & Diamant 2004), zebrash (Danio
rerio) (Kimmel et al. 1995) and black skirt tetra
(present study). The timing of ination in black
skirt tetra larvae was earlier than in many other
species; for example, in discus, D. Rerio, O. mos-
sambicus, Stizostedion vitreum, Latris lineata, swim
bladder inates at 45 DAH, 5 DAH, 79 DAH,
612 and 1120 DAH respectively (Boulhic &
Gabaudan 1992; Marty et al. 1995; Trotter et al.
2001; O

nal et al. 2010). In contrast, ination in

freshwater angelsh occurred at 12 DAH (Zilberg
et al. 2004).
In the present study, the morphological develop-
ment and allometric growth patterns in the black
skirt tetra larvae were studied. The allometric
growth formula is the most widely used method
of analysis for relative growth during early
larval development (Osse & van den Boogaart
2004; Pen a & Dumas 2009). Teleostean larval
period was characterized by a high degree of allo-
metric growth patterns (Fuiman 1983; Osse, van
den Boogaart, van Snik & van der Sluys 1997;
Geerinckx, Verhaegen & Adriaens 2008). These
patterns can contribute to aquaculture and sher-
ies management by characterizing normal growth
patterns (Pen a & Dumas 2009). Allometric
growth during larval development was studied in
different teleost groups (Osse & van den Boogaart
2004). But allometric growth of many ornamental
sh has not been reported. We described the allo-
metric growth patterns of G.ternetzi larvae from
hatching to day 32. The growth coefcients of
head, trunk and tail during larval periods (Period
I: yolk-sac larva, period ii: preexion larva, period
iii: exion larva, period iv: postexion larva) were
similar to results that have been reported in
other teleosts (Osse & van den Boogaart 1999;
Geerinckx et al. 2008; Huysentruyt, Moerkerke,
Devaere & Adriaens 2009). The allometric changes
chronology would be related to the chronology of
important early life history events (Huysentruyt
et al. 2009) and positive allometry of head and tail
length during this period reects the early priority
to develop organs related to vital functions such as
feeding and swimming (Osse & van den Boogaart
2004).
In conclusion, our ndings indicated that black
skirt tetra eggs are demersal, adhesive and their
larvae are altricial. Hatching occurred after an
incubation period of 2022 h at 24 0.5C.
The cleavage pattern of G.ternetzi is the same as
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1271
Aquaculture Research, 2012, 43, 12601275 Embryonic and larval development of black skirt tetra I

C elik et al.
those of other characin (Romagosa, Narahara &
Fenerich-Verani 2001; dos Anjos & dos Anjos
2006; Pan, Zhan & Gong 2008; Faustino, Nakaghi
& Neumann 2010) and zebrash Danio (Brachyda-
nio) rerio (Kimmel et al. 1995). Egg hatching time
of black skirt tetra is different from that of others.
Early larval development of G.ternetzi was divided
into four different periods: Yolk-sac larva; the pres-
ence of a yolk sac ventrally in the body, between
hatching and 4 DAH. Yolk sac was absorbed and
larvae swim actively 34 days after hatching and
the onset of exogenous feeding occurred 3 days
later. Preexion larva: this period begins at absorp-
tion of yolk sac and ends at the start of upward
exion of the notochord (between 4 and 11 DAH).
Flexion larva: this period (the period during noto-
chord exion) is characterized with the hypural
bones assuming a vertical position, between 11
and 12 DAH. Postexion larva: the period between
completion of exion and the juvenile stage,
1232 DAH. Our ndings may provide a basis for
further studies to determine the complete early life
history of black skirt tetra G. ternetzi and detailed
studies such as these may be helpful for commer-
cial production of some ornamental sh. The
results of this study can contribute to a better
understanding of the embryonic and larval devel-
opment of other commercial characin larvae. They
can be used to explain some aspects of the early
life history at culture conditions and can help to
develop better larval culture methodologies in
hatchery. Similarly, they will be helpful to increase
success rates in the larval culture of some freshwa-
ter ornamental sh species. New approaches can
be brought to solve many problems in larval cul-
ture of many other species.
References
Alves M.S.D. & Moura A. (1992) Estadios de desenvolvi-
mento embrionario de curimata-pioa Prochilodus afnis
(Reinhardt, 1874) (Pisces, Prochilodontidae) em 1992.
In: Encontro Anual de Aquicultura de Minas Gerais Belo
Horizonte. Anais, pp. 6171. Codevasf , Belo Horizon-
te, Tres Marias.
Andrade-Talmelli E.F., Kavamoto E.T., Romagosa E. &
Fenerich-Verani N. (2001) Embryonic development of
the piabanha, Brycon insignis, (Steindachner, 1876)
(Pisces; Characidae). Boletim do Instituto de Pesca 27,
2128.
Arezo M.J., Pereiro L. & Berois N. (2005) Early develop-
ment in the annual sh Cynolebias viarius. Journal of
Fish Biology 66, 13571370.
Arvedlund M., McCormick M.I. & Ainsworth T. (2000)
Effects of photoperiod on growth of larvae and juve-
niles of the anemonesh Amphiprion melanopus. Naga,
The ICLARM Quarterly 23 (2), 1823.
Ballard W.W. (1986) Morphogenetic movements and a
provisional fate map of development in the Holostean
sh Amia cavla. Journal of Experimental Zoology 238,
355372.
Bermudes M. & Ritar A.J. (1999) Effects of temperature
on the embryonic development of the striped trumpeter
(Latris lineata Bloch and Schneider, 1801). Aquacul-
ture176, 245255.
Bhattacharya H., Zhang S.C. & Wang Y.J. (2005) Embry-
onic development of the rosy barb Puntius conchonius
Hamilton 1822 (Cyprinidae). Tropical Zoology 18 (2),
2537.
Bjelland R.M. & Skiftesvik A.B. (2006) Larval develop-
ment in European hake (Merluccius merluccius L.)
reared in a semi-intensive culture system. Aquaculture
Research 37, 11171129.
Borcato F.L., Bazzoli N. & Sato Y. (2004) Embriogenesis
and larval ontogeny of the Pirugordura,Leporinus piau
(Fowler) (Pisces, Anostomidae) after induced spawning.
Revista Brasileira de Zoologia 21, 117122.
Borges R., Faria C., Gil F., Goncalves E.J. & Almada V.C.
(2003) Embryonic and larval development of Gobius
paganellus (Pisces: Gobiidae). Journal of the Marine Bio-
logical Association of the U.K. 83, 11511156.
Boulhic M. & Gabaudan J., (1992) Histological study of
the organogenesis of the digestive system and swim-
bladder of the Dover sole, Solea solea (Linnaeus 1758).
Aquaculture 102, 373396.
Britz R. (1997) Egg surface structure and larval cement
glands in nandid and badid shes with remarks on
phylogeny and biogeography. American Museum Novi-
tates 3195, 117.
C elik I

. (2008) Diskus balklarnda uremeye etki eden

faktorlerin belirlenmesi ve larva/juvenil gelisim ozellikleri.
C anakkale Onsekiz Mart U

niveristesi, Fen Bilimleri

Enstitu su , C anakkale Onskiz Mart University Pub.,
C anakkale/Turkey, PhD Thesis, 97pp.
Cole B. & Haring M. (1999) Spawning and Production of
the Serpae Tetra, Hyphessobrycon serpae. Center for Trop-
ical and Subtropical Aquaculture Publication Number
138, Hawaii.
Coleman R.M. & Galvani A.P. (1998) Egg Size Deter-
mines Offspring Size in Neotropical Cichlid Fishes (Tele-
ostei: Cichlidae). American Society of Ichthyologists and
Herpetologists 1, 209213.
Du R., Wang Y., Jiang H., Liu L., Wang M., Li T. &
Zhang S. (2010) Embryonic and larval development in
barn ounder Verasper moseri (Jordan and Gilbert).
Chinese Journal of Oceanology and Limnology 28(1),
1825.
dos Anjos H.D.B. & dos Anjos C.R. (2006) Biologia
reprodutiva e desenvolvimento embrionaro e larval
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1272
Embryonic and larval development of black skirt tetra I

C elik et al. Aquaculture Research, 2012, 43, 12601275

do cardinal tetra, Paracheirodon axelrodi Schultz, 1956
(Characiformes: Characidae), em laboratorio. Boletim do
Instituto de Pesca, Sao Paulo 32 (2), 151160.
Faustino F., Nakaghi L.S.O. & Neumann E. (2010) Bry-
con gouldingi (Teleostei, Characidae): Aspects of the
embryonic development in a new sh species with aquacul-
ture potential Zygote: pp. 113. Cambridge University
Press, doi:10.1017/S0967199410000535.
Finn R.N., Rnnestad I., van der Meeren T. & Fyhn H.J.
(2002) Fuel and metabolic scaling during the early life
stages of Atlantic cod Gadus morhua. Marine Ecology
Progress Series 24, 217234.
Frankel J.S. (2004) Inheritance of Trunk Banding in the
Tetra (Gymnocorymbus ternetzi Characidae). Journal of
Heredity 95 (2), 262264.
Fuiman L.A. (1983) Growth gradients in sh larvae.
Journal of Fish Biology 23, 117123.
Fukuhara O. (1990) Effects of temperature on yolk utili-
zation, initial growth, and behaviour of unfed marine
sh-larvae. Marine Biology 106 (2), 169174.
Geerinckx T., Verhaegen Y. & Adriaens D. (2008) Onto-
genetic allometries and shape changes in the sucker-
mouth armoured catsh Ancistrus cf. triradiatus
Eigenmann (Loricariidae, Siluriformes), related to suck-
ermouth attachment and yolk-sac size. Journal of Fish
Biology 72, 803814.
Gisbert E., Merino G., Muguet J.B., Bush D., Piedrahita R.
H. & Conklin D.E. (2002) Morphological development
and allometric growth patterns in hatchery-reared
California halibut larvae. Journal of Fish Biology 61,
12171229.
Govoni J.J. & Forward R.B Jr (2008) Buoyancy. In: Fish
Larval Physiology (ed. by R.N. Finn & B.G. Kapoor), pp.
495522. Science Publishers, Eneld, NH, USA.
Hall T.E. (2008) Pattern formation In: Fish larval physiol-
ogy, (eds.R.N. Finn & B.G. Kapoor) pp. 325. Science
Publishers, Eneld, New Hampshire, USA.
Hall T.E., Smith P. & Johnston I.A. (2004) Stages of
embryonic development in the Atlantic cod Gadus mor-
hua. J. Morphology 259, 255270.
Hanel R., Karjalainen J. & Wieser W. (1996) Growth of
swimming muscles and its metabolic cost in larvae of
whitesh at different temperatures. Journal of Fish Biol-
ogy 48, 937951.
Hunt von Herbing I. & Boutilier R.G. (1996) Activity and
metabolism of larval Atlantic cod (Gadus morhua)
from Scotian Shelf and Newfoundland source popula-
tions. Marine Biology 124, 607617.
Huysentruyt F. & Adriaens D. (2005) Adhesive structures
in the eggs of Corydoras aeneus (Gill, 1858; Callich-
thyidae). Journal of Fish Biology 66, 871876.
Huysentruyt F., Moerkerke B., Devaere S. & Adriaens D.
(2009). Early development and allometric growth in
the armoured catsh Corydoras aeneus (Gill, 1858).
Hydrobiologia 627, 4554.
Iwamatsu T. (1994) Stages of normal development
in the medaka Oryzias latipes. Zoological Science 11,
825839.
Kendall A.W., Ahlstrom E.H. & Moser H.G. (1984). Early
life history stages of shes and their characters. In
Ontogeny and systematics of shes: American Society of
Ichthyologists and Herpetologists, Special Publication
No. 1 (ed. by Moser H.G., Richards W.J., Cohen D.M.,
Fahay M.P., Kendall A.W. & Richardson S.L.), pp. 11
22. Allen Press Inc, Lawrence, Kansas, U.S.A.
Khan M.M.R. & Mollah M.F.A. (1998) Embryonic and
larval development of African catsh, Clarias gariepinus
(Burchell). Bangladesh Journal of Fisheries 21, 9197.
Kimmel C.B., Ballard W.W., Kimmel S.R., Ullman B. &
Schilling T.F. (1995) Stages of embryonic development
of the zebrash. Developmental Dynamics 203, 253310.
Koumoundouros G., Divanach P. & Kentouri M. (2001)
Osteological development of Dentex dentex (Osteichth-
yes:Sparidae): dorsal, anal, paired ns and squamation.
Marine Biology 138, 399406.
Marimuthu K. & Haniffa M.A. (2007) Embryonic and lar-
val development of the striped snakehead Channa stria-
tus. Taiwania 52 (1), 8492.
Martell D.J., Kieffer J.D. & Trippel E.A. (2005) Effects of
temperature during early life history on embryonic
and larval development and growth in haddock.
Journal of Fish Biology 66, 15581575.
Martinez G.M. & Bolker J.A. (2003) Embryonic and stag-
ing of summer ounder (Paralichthys dentatus). Journal
of Morphology 255, 162176.
Marty G.D., Hinton D.E. & Summerfelt R.C. (1995) Histo-
pathology of swimbladder nonination in walleye (Stiz-
ostedion vitreum) larvae: role of development and
inammation. Aquaculture 138, 3548.
Meijide F.J. & Guerrero G.A. (2000) Embryonic and
larval development of a substrate-brooding cichlid
Cichlasoma dimerus (Heckel, 1840) under laboratory
conditions. Journal of Zoology 252 (4), 481493.
Mommsen T.P. (2001) Paradigms of growth in sh.
Comparative Biochemistry and Physiology B 129, 207
219.
Morioka S., Ito S. & Kitamura S. (2010) Growth and
morphological development of laboratory-reared larval
and juvenile snakeskin gourami Trichogaster pectoralis.
Ichthyological Research 57, 2431.
Moyle P.B. & Cech J.J. (2000) Fishes: An Introduction to
Ichthyology, Fourth Edition. Prentice-Hall, Upper Saddle
River, NJ, 6978.
Nakatsuji T., Kitano T., Akiyama N. & Nakatsuji N.
(1997) Ice goby (Shiro-uo), Leucopsarion petersii, may
be a useful material for studying teleostean embryo-
genesis. Zoological Science 14, 443448.
Nandini S. & Sarma S.S.S. (2000) Zooplankton prefer-
ence of two species of freshwater ornamental sh
larvae. Journal of Applied Ichthyology 16, 282284.
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1273
Aquaculture Research, 2012, 43, 12601275 Embryonic and larval development of black skirt tetra I

C elik et al.
Nelson J.S. (1994) Fishes of the world. Third edition. John
Wiley & Sons, Inc., New York. 600 p. Characidae).Jour-
nal of Heredity 95 (3), 262264.
Olivotto I., Holt S.A., Carnevali O. & Holt G. J. (2006)
Spawning, early development, and rst feeding in the
lemonpeel angelsh Centropyge avissimus. Aquaculture
253, 270278.
Osse J.W.M., van den Boogaart J.G.M., van Snik G.M.J. &
van der Sluys L. (1997) Priorities during early growth
of sh larvae. Aquaculture 155, 249258.
Osse J.W.M. & van den Boogaart J.G.M. (1999) Dynamic
morphology of sh larvae, structural implications of
friction forces in swimming, feeding and ventilation.
Journal of Fish Biology 55 (Supplement A), 156174.
Osse J.W.M. & van den Boogaart J.G.M. (2004) Allo-
metric growth in Fish Larvae: Timing and Function.
In: The Development of Form and Function in Fishes and
the Question of Larval Adaptation, (ed. by J.J. Govoni),
pp. 167194. American Fisheries Society, Symposium
40, Bethesda, MD, USA.
O

nal U., C elik I

. & Cirik S. (2010) Histological develop-

ment of digestive tract in discus, Symphysodon spp. Lar-
vae. Aquaculture International 18, 589601.
Pan X., Zhan H. & Gong Z. (2008) Ornamental expres-
sion of red uorescent protein in transgenic founders
of white skirt tetra (Gymnocorymbus ternetzi). Marine
Biotechnology 10, 497501.
Papoutsoglou E.S. & Lyndon A.R. (2005) Effect of incu-
bation temperature on carbohydrate digestion in
important teleosts for aquaculture. Aquaculture
Research 36, 12521264.
Pelster B. (2004) The development of the swim bladder:
structure and performance. In: The Development of Form
and Function in Fishes and the Question of Larval Adapta-
tion (ed. by J.J. Govoni), American Fisheries Society Sym-
posium 40, 3746. Bethesda.
Pen a R. & Dumas S. (2009) Development and allomet-
ric growth patterns during early larval stages of the
spotted sand bass Paralabrax maculatofasciatus (Percoi-
dei: Serranidae). Scientia Marina 73(Special issue), 183
189.
Perlberg S.T., Diamant A., Or R. & Zilberg D. (2008)
Characterization of swim bladder non-ination (SBN)
in angelsh, Pterophyllum scalare (Schultz), and the
effect of exposure to methylene blue. Journal of Fish
Diseases 31, 215228.
Rahman M.R., Rahman M.A., Khan M.N. & Hussain M.
G. (2004) Observation on the embryonic and larval
development of silurid catsh, gulsha (Mystus cavasius
Ham). Pakistan Journal of Biological Sciences 7 (6),
10701075.
Reynalte-Tataje D., Zaniboni-Filho E. & Esquivel J.R.
(2004) Embryonic and larvae development of pira-
canjuba, Bryconorbignyanus Valenciennes, 1849 (Pisces,
Characidae). Acta Scientiarum. Biological Sciences Maringa
26 (1), 6771.
Richards W. J. & Leis J. M. (1984) Labroidei: develop-
ment and relationships. In Ontogeny and Systematics of
Fishes: American Society of Ichthyologists and Herpetolo-
gists. Spec. Publ. No. 1, (ed. by Moser H.G., Richards
W.J., Cohen D.M., Fahay M.P., Kendall A.W. & Rich-
ardson S.L.), pp. 542547. Allen Press Inc, Lawrence,
Kansas, USA.
Romagosa E., Narahara M.Y. & Fenerich-Verani N.
(2001) Stages of embryonic development of the
Matrnxa, Brycon cephalus (Pisces, Characidae). Bole-
tim do Instituto de Pesca, Sao Paulo 27 (1), 2732.
Saillant E., Chatain B., Fostier A., Przybyla C. & Fauvel C.
(2001) Parental inuence on early development in the
European sea bass. Journal Fish Biology 58, 15851600.
Savas E., Sener E. & Yldz M. (2006) Japon balklarnda
(Carassius sp.) embriyolojik ve larval gelismenin ince-
lenmesi. I

stanbul Universitesi Veterinerlik Faultesi Dergisi

32 (3), 719.
Silva L.V.F. (2004) Morphology and early development of
silver catsh, Rhamdia quelen, (Siluriforme, Pimelodi-
dae) embryos and larvae. In: Physiology of Fish Eggs
and Larvae Symposium Proceedings, ( ed. by D. MacKin-
lay), 8994. International Congress on the Biology of
Fish Manaus, Brazil.
Stiassny M.L.J. & Mezey J.G. (1993) Egg attachment sys-
tems in the family Cichlidae (Perciformes: Labroidei),
with some comments on their signicance for phyloge-
netic studies. American Museum Novitates 3058, 111.
Takeshita N., Onikura N., Matsui S. & Kimura S. (1997)
Embryonic, larval and juvenile development of the
roughskin sculpin, Trachidermus jasciatus (Scorpaenifor-
mes: Cottidae). Ichthyological Research 44 (3), 257
266.
Tamaru C.S., Cole B., Bailey M.S.R. & Brown B.A.C.
(2001) A manual for commercial production of the tiger
barb, Capoeta tetrazona, a temporary paired tank spawner.
Center for Tropical and Subtropical Aquaculture Publi-
cation Number 129. Hawaii.
Trotter A.J., Pankhurst P.M. & Hart P.R. (2001) Swim-
bladder malformation in hatchery-reared striped trum-
peter Latris lineata (Latridae). Aquaculture 198, 4154.
Trotter A.J., Pankhurst P. M. & Battaglene S.C. (2004)
Morphological development of the swim bladder in
hatchery-reared striped trumpeter Latris lineata. Journal
of Applied Ichthyology 20, 395401.
Uma B. & Chandran M.R. (2008) Induction of Triploidy
in Gymnocorymbus Ternetzi (Boulenger). Research Jour-
nal of Fisheries and Hydrobiology 3 (2), 4147.
U

nal H. & Aral O. (2006) C opcu balklar (Corydoras

spp.) ve yetistiriciligi. Journal of Fisheries and Aquatic
Sciences 23(1-2), 311318.
Watson C.A., ChapmanF.A. (2002). Articial incubation of
sh eggs. Fact Sheet FA-32, Institute of Food and
Agricultural Science, University of Florida Exten-
sion. Available at http://edis.ifas.u.edu/pdfles/FA/
FA05100.pdf (accessed 03 February 2011).
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1274
Embryonic and larval development of black skirt tetra I

C elik et al. Aquaculture Research, 2012, 43, 12601275

Webb J. (1999)Larvae in sh development and evolution.
In The Origin and Evolution of Larval Forms(ed. by B.K.
Hall & M.H. Wake), pp. 109158. Academic Press,
San Diego, CA, USA.
Wood C.M. & Mc Donald D.G. (1996) Global warning:
Implications for freshwater and marine sh. Society for
Experimental Biology Seminar Series 61. Cambridge Uni-
versity Press, Cambridge, pp. 177223.
Zilberg D., Or R., Rabinski T. & Diamant A. (2004)
Morphological and genetic characterization of swim-
bladder non-ination in angelsh Pterophyllum scalare
(Cichlidae). Aquaculture 230, 1327.
2011 Blackwell Publishing Ltd, Aquaculture Research, 43, 12601275 1275
Aquaculture Research, 2012, 43, 12601275 Embryonic and larval development of black skirt tetra I

C elik et al.