(Amphiprion perideraion)

J. Fish. Soc. Taiwan, 35(1): 75-85
Embryo Development and Larval Rearing of Pink Clownfish
(Amphiprion perideraion)
Yuan-Shing H0 1, Che-Ming Chen', Wen-Vie Chen' and Wen-Been Chang 2 ,3*
(Received, February 1, 2008; Accepted, March 15, 2008)
ABSTRACT
Pink clownfish (Amphiprion perideraion) belongs to the subfamily of Amphiprioninae in the
Pomacentridae family. During the 247 days of observation , the pink clownfish ovulated 21 times. The
oviposition came after the brooders began to clean the spawning substrate together ; the venter of female
grew larger with a protrusion in the female breed aperture. The spawning interval and hatching time required
of A. peride raion under 27 ± 1DC of water temperature and 34 ± 1 parts per thousand (ppt) of salinity was
13.0 ± 3.7 day & 7.2 ± 1.2 (n = 49 ) days post spawning (DPS) , with about 300-700 eggs per spawning.
The spawning activity lasted more than one hour starting from 09 : 15 to 14 : 05 with most spawning from
09 : 00 to 11 : 00. These adhesion demersal eggs were orange and ellipsoidal, which were about 1.85 ±
0.20 (n = 30) mm in diameter. The yolks were about 1.30 ± 0.13 mm in diameter and contained several
oil droplets were about 152 ± 109 urn in diameter. The newly hatched larvae were about 3.20-3.80 mm in
total length, which were phototactic and the phototaxis decreased as they grew. The larvae were fed with
rotifers (Brachionus plicatilis) from day one post hatching (1 DPH) to 11 DPH. On 12 DPH , copepod was
added to the feeds and the amount was increased gradually until it totally replaced rotifers on the 15 DPH .
The 22 DPH fry showed fighting and territorial behavior. The band on the body surface of the larvae at 26
DPH (12.56 mm) is similar to that of adult. The 59 DPH old fry were about 16.62-28.63 mm in total length
and could be completely fed with pellet feed . The development of protective systems in those without any
changes in color is incomplete .
Key words: Amphiprion perideraion, Embryo development, Larval rearing .
INTRODUCTION
Tropical marine ornamentals comprise
an increasingly important fishery worldwide. It has been shown that both
anemone and anemonefish densities were
significantly lower in exploited areas than
in protected areas, and anemonefish and
anemones comprised close to 60% of the
total ornamental fish catch in Philippines
(Shuman et al ., 2005). Fishes on coral
reefs are characterized by large fluctuations
in recruitment (Doherty, 1988; Sale et al. ,
1994). These fluctuations in cohort strength
may be governed in part by environmental
1 Eastern
conditions affecting growth and development of larvae (Meekan & Fortier, 1996 ;
Theilacker et al., 1996). As a result of such
mechanisms acting singly or in conjunction,
a critical period or catastrophic mortality
stage can be identified in the reproductive
cycle of marine fishes which chronologically
correlates very closely with the period shortly
after the yolk sac has been fully absorbed.
The recruitment of the vast majority of coral
reef fishes relies on the inflow of a pelagic
larval stage (Leis, 1991) with the larval
distribution range dependent on several
factors including water current and larval life
span. The tropical marine anemonefishes
Marine Biology Research Center, Fisheries Research Institute
of Marine Biodiversity and Evolution, National Dong Hwa University
3 National Museum of Marine Biology and Aquarium
• Corresponding author, E-mail: [email protected]
2 Institute
TAO
76
Yuan-Shing Ho, Che-Ming Chen, Wen-Vie Chenand Wen-Been Chang
(Pomacentridae) are important in the trade
for ornamental fish (Wilkerson, 1998).
Clownfish are one of the easier tropical
marine aquarium fish to breed. Some
mariculture centers and scientific laboratories
have started rearing these anemonefish
(McLarney 1985, 1986; Miyagawa, 1989;
Hoff, 1993; Young, 1996; Job et al., 1997).
Unlike many of the other tropical marine
aquarium fish , clownfish regularly spawn
in marine aquarium. Pink clownfish
(Amphiprion perideraion) , found in the
depths from 3 m to 30 m, inhabits coral reefs
and is usually associated with the anemone
Heteractis magnifica sometimes seen
associated with other anemones, Heteractis
crispa and Stichodactyla gigantea , has
been exploited commercially for the growing
marine aquarium trade. In heavily exploited
areas, replenishment may be dependent
on spawning from elsewhere. Amphiprion
spp. have a short larval period (15-20 days)
relative to other coral reef fishes (Wellington
& Victor, 1989). The consequential reduction
in stock size may increase the risk of stock
fluctuations and render them vulnerable to
over exploitation. The purpose of this article
is to convey the information that we have
found and learned a few very important
steps to breeding , spawning, and larval
rearing technique of c1ownfish. Studies on
the early ontogeny of A. perideraion not only
contribute to the existing knowledge about
the developmental features of this species,
but also establish a model for comparison
when normal patterns of the embryonic
development are altered.
MATERIALS AND METHODS
Eight mature brood stock clownfish
(Amphiprion perideraion) and several
radiation sea anemone (Radianthus ritten)
were raised in a 400 L FRP tank. The
tank was maintained at 27 ± 1DC of water
temperature and 34 ± 1 parts per thousand
(ppt) of salinity. We used a halogen lamp
to illuminate the tank for 9 hours/day. The
broodstock were fed fresh and frozen shrimp,
squid, fish and artificial granular fodder
daily. Behavior mode and time sequence
before and after spawning of clownfish
were recorded using a digital camera. The
number of eggs was estimated after the end
of each spawning.
After spawning , the diameter of fertilized
eggs and oil globule were measured by a
microscope with a 40x magnification. A total
of 10 adhesive eggs were sampled randomly
at each sampling interval. The eggs were
transferred into sterile vessels disinfected
with UV light. The sterile vessels were filled
with filtered seawater. The eggs need to be
constantly agitated by a gentle flow of water.
Each egg was placed on a slide to observe
the embryonic morphological development.
Time was recorded when 50% of the
embryos attained cleavage, blastulation ,
gastrulation, organogenesis , embryo , prehatching and larval stage clearly. The main
morphological and functional features of
each development stage were recorded.
Newly-hatched larvae with phototaxis will
swim towards the upper part of the tank that
we can collection and raises them. Larvae
on different periods were fed with rotifer
(Brachionus plicatilis), copepod , nauplii of
Artemia and artificial feedstuff, respectively.
Digital camera with a stereomicroscope
can take the shape change of fish larvae
fin, body length and body color on different
growth phases, periodically.
30 larvae with change in color and 30
larvae without any change in color (17 day
post hatching, DPH) with a radiation sea
anemone (R. ritten) put into 1000 ml beaker
with 500 ml fresh sea water, respectively. It
let larvae easy to touch the sea anemone at
any time. The survival rate of fish larvae was
recorded after 2 hours and 24 hours.
TAO
RESULT
Among the 8 adult fish only three
survived and two of them became a pair.
The total length and weight of matured
males and females are 73 .8 mm, 7.5 g
and 68.7 mm , 6.4 g , respectively. The
brood stock of pink clownfish (Amphiprion
perideraion) was fed with fresh and frozen
shrimp, squid , fish and artificial granular
fodder. Before spawning commences the
Reproductionof Pink Clownfish
fish will go through some sort of courtship .
This will probably be displayed in head
wagging , shimmering and biting. There is
considerable "interplay" between the male
and female prior to spawning, the male fish
draw the female to the spawning surface and
encourage her to clean, by biting to remove
any algae or debris, from the proposed
spawning site and keep clean until spawning
(Figure 1). The first spawn took place on
March 17,2003. When spawning begins ,
the female will lay a line of eggs and then
the male will immediately follow and fertilize
them.
The spawning interval of A. perideraion
was 13 .0 ± 3 .7 (n = 49) days under a
seawater temperature of 27 ± 1°C and salinity
of 34 ± 1 ppt. Pink clownfish spawned about
300-700 eggs in total per spawning in this
study, and the spawning activity lasted more
77
than an hour, which usually started from 09
: 15 to 14 : 05 with most spawning occurred
from 09 : 00 to 11 : 00. The hatching time
required for A. perideraion were 7.2 ± 1.2 (n
49) day post spawning (DPS).
Once the clownfish started spawning,
they are likely to repeat at intervals of around
10 to 15 days mostly, usually takes place
between 9 and 11 am, and spawns a total
of 21 times (300-700 eggs/ spawn) from
March 17 to November 13, 2003. The
parent fish usually perform this function by
fanning the eggs with their pectoral fins.
The constant movement prevents clumping
and smothering , which makes the eggs
unsusceptible to fungus and bacterial attack,
and ensures that the eggs are well aerated.
The deposited eggs were orange , prolate
spherical and adhesive, with a yolk of about
1.13-1.45 mm in diameter, which contained a
=
TAO
Fig. 1. Breeding activit ies of Amphiprion perideraion . A: The pair of brooder we re cleaning the spawning
substrate together ; B: Female in spawning ; C: Ejaculating male; D: Fertilized eggs were guarded by
brooders.
Yuan-Shing Ho, Che-Ming Chen, Wen-VieChen and Wen-Been Chang
78
large oil globule (152 ± 109 IJm in diameter).
The size of yolk reduced gradually when the
cleavage proceeds as the same basic pattern
in teleost fish . The fertilized eggs looked
like little capsules about 1.85 ± 0.20 (n 30)
mm long in diameter. The yolks were about
1.30 ± 0.13 mm in diameter and contained
several oil droplets were about 152 ± 109
IJm in diameter. Embryonic development
of A. perideraion from 0 h to 24 h after
insemination is considered as the cleavage
phase. The embryon ic development is
summarized in Figure 2. The egg indicated a
typical discoidal cleavage, creating a cellular
=
c
B
A
region at the posterior of the yolk mass.
The second cleavage occurred a asingle
meridional plane at right angle to the first ,
giving rise to 4 blastomeres 1 h 32 min after
insemination (Fig. 2, A). The egg attained
the 8 blastomere stage at 2 h (Fig . 2, B).
The embryo of A. perideraion took 2 h 25
min, 3 h, 3 h 30 min, 4 h 20 min, 16 h 45 min
to reach 16, 32, 64-blastomere, morula stage
and gastrula stage (Fig. 2, C-G), respectively.
The blastomeres roundish mound perched
on the top of the yolk in the figures (Fig. 2,
F). Gastrulation process is characterized
as different morphogenetic movements for
n
E
b
.J
op
og
F
G
L
TAO
o
h
Fig. 2. Embryonic development of Amphiprion perideraion. A: 4-blastomere stage; B: 8- blastomere stage; C:
16-blastomere stage; D: 32-blastomere stage; E: 64-blastomere stage; F: Morula Stage; G: Gastrula
stage; H: 2/3 of yolk was covered with blastodisc, embryo appeared; I: Optic vesicles appeared, 5
somites; J: Auditory vesicles formed, 14 somites; K: Optic lens and tail formed, tail freed from yolk sac,
chromatophore on embryo and yolk, 16 somites; L: Embryo moved spastically, heart rate: 100-104 times/
min, 26 somites; M: Head of embryo move to the top of egg, 28 somites; N: Chromatoplasm precipitated
on eyes, heart rate: 156 times/min, 30 somites; 0: The end of tail reached eyes, pectoral fin swung
sometime, Guanine accumulated on eyes, and heart rate: 176-180 times/min. (b: blastomeres; bp: body
pigment; ep: eye pigment; es: embryonic-shield; ez: evacuation-zone; h: heart; hb: hindbrain; I: lens; og:
oil globule; op: otic placode; t: tial; y: yolk).
Reproduction of PinkClownfish
the required occurrence of epiboly. Epiboly
began 16 h 45 min after insemination, when
the blastomeres covered one-quarter of
the egg. The epiboly was evident at the
animal pole forming a cap-like structure. The
thickened part of the blastomeres became
an embryonic shield (Fig. 2, G). Embryonic
body formation was observed when the
blastomeres covered the whole yolk and
formed the notochord at 23 h 10 min (Fig. 2,
H). After blastopore closed the yolk mass ,
the body of A. perideraion embryo was
formed and tail-bud appeared at the posterior
of the yolk mass . Notochord appeared
with the head and eye directed towards the
attached end of the egg . This transformation
is called organogenesis ; auditory vesicles
took place , and embryo reached the
5-somite stage at 27 h 48 min (Fig. 2, I). The
embryo reached the 14-somite stage at 33 h
20 min (Fig. 2, J). The embryo reached the
16-somite at 38 h 32 min with the formation
of optic lens and tail; the tail freed from yolk
sac and the chromatophore was visible on
embryo and yolk (Fig. 2, K). Embryo moved
spastically, the heart was cup-shaped with
pink-red in color and the pulsation reached
100-104 times/min , and the embryo reached
the 26-somite at 43 h 20 min (Fig. 2, L). on
56 h after fertilization , the head of embryo
turned to the top of egg, 28-somite appeared
(Fig. 2, M); on 78 h 30 min after fertilization,
chromatoplasm precipitated on eyes , and
heartbeat rate became 156 times/min,
30-somite appeared (Fig. 2 , N); Pectoral
fin became functional while the head and
tail moved actively. The end of tail reached
eyes, Guanine accumulated on eyes , and
the heart rate was 176-180 times/min on
day 5 (115 h 11 min) after insemination (Fig.
2, 0). Comparative the iridescent metallic
silver eye color implied that the embryo was
getting to be hatched. Such color changes
also indicate that the embryo development
is healthy. The newly hatched larvae with
26-somite, 3.20-3.80 mm in total length and
the heart heartbeat rate became 180-200
times/min , occurred at 151 h 20 min (Fig. 2,
P), which is phototactic and the phototaxis
lessened as they grew.
The first day post hatching (DPH) larvae
79
(Fig. 3 , A) and just before the mouth of
the larvae opens, rotife rs (7-13 ind/ml) are
distributed. The caudal fin began forming
and the pectoral fin developed in round
shape as fin fold at average body length 3.55
mm and swim towards the lower part of the
tank by the end of 2 DPH (Fig. 3, B). A mode
of 15 dorsal soft rays, 12 anal soft rays and
19 caudal soft rays has developed at 4 DPH.
Larval meristic data as D X,16; A II, 13;
Vertebrae 14 has began forming between 6
and 12 DPH. The fin development observed
at this stage was already similar to that of
an adult. Clownfish larvae (Fig . 3, C-D)
with black or dark gray body color but light
silvery abdomen was observed between 8
DPH (4.47 mm) and 10 DPH (6 .22 mm).
The larvae's body color changes into pink
(Fig. 3, E) at 12 DPH (7.13 mm) and tries to
feed copepod at this stage . Thin white lines
appeared on the head top of larvae at 15
DPH and can feed full copepod at this stage.
The thin white lines also appeared on the
base of dorsal fin and connect with those on
the head top of larvae (Fig. 3, F) at 22 DPH
(10.44 mm). A thin narrow and perpendicular
white band appeared on the rear of the
head (Fig. 3, G) , and the larvae fought for
territories at 22 DPH (10.93 mm). The band
on the body surface of the larvae at 26 DPH
(12.56 mm) is similar to that of adult. The
larva has already accepted artificial granular
fodder completely at 59 DPH (16.62-28.63
mm) (Fig. 3, H).
Larvae in the culture tank were fed with
rotifers (Brachionus plicati/is) from 1-11 DPH.
Copepod were introduced to the tank at 12
DPH for 48 days and increased gradually
until it totally replaced rotifers at the 15th day
(Fig. 4). Subsequently, artificial feedstuff was
fed to the larvae at 59 DPH (16.62-28.63
mm).
The mortality ratio of turned coloration
and didn't coloration of A. perideraion larvae
(17 DPH) which were killed by radiation sea
anemone (R. rittert) in 2 hours & 24 hours
were 3 (10.0%) & 4 (13.3%) and 27 (10.0%)
& 30 (13 .3%) , respectively. The mortality
ratio of A. perideraion larvae with change
in color and without any change in color
(17 DPH) in the present study has showed
TAO
80
Yuan-Shing Ho, Che-Ming Chen , Wen-Vie Chen and Wen-Been Chang
TAO
Fig. 3. Morphological changes of the Amphiprion perideraion at fry and larval stage. A: Newly-hatched larva,
3.08 mm in total length; B: 2 DPH, 3.55 mm in total length; C: 8 DPH, 4.47 mm in total length ; D: 10 DPH ,
6.22 mm in total length; E: 12 DPH , 7.13 mm in total length ; F: 22 DPH , 10.44 mm in total length ; G: 24
DPH, 10.93 mm in total length; H: 26 DPH, 12.56 mm in total length.
Reproduction of PinkClownfish
81
50
45
40
35
""""
E
E
'-'
..c
......
eo
l:
~
-a......
0
r-
30
25
20
15
Rotifer
10
Pe llet feed
5
0
0
30
60
90
120
150
180
210
240
Age (Days)
Fig. 4. Growth performance of Amphiprion perideraion fed different diets.
that the development of protective systems
in those without any changes in color is
incomplete.
DISCUSSION
The protandrous anemonefish often
forms a group consisting of a large female ,
a small male, and a smaller nonbreeder at
an isolated single host anemone , where the
home ranges of subordinates were covered
with the female's home range. With in the
group, the dominant individuals suppress the
growth of subordinates , resulting in large size
differences (Kobayashi and Hattori , 2006).
Female Amphiprion perideraion laid demersal
eggs on rocky nest sites which are protected
by males. Theses males guard the fertilized
eggs from predators until the embryonic fish
hatch out as pelagic larvae . Pigmentation
of newly deposited egg depends primarily
on species and parental nutrition (Allen ,
1974; Swanson, 1996; Wilkerson, 2001;
Arezo et a/., 2005; Kovac, 2005). The yolk of
clownfish was bright in color, tinted with the
parent color pigment. It has been recorded
that Clark clown, Pinks clown and Maroons
clown eggs are yellowish-orange, pinkish
and red , respectively (Wilkerson , 2001).
These findings are in agreement with the
results obtained in the present study, which
show that the newly deposited Amphiprion
perideraion eggs are orange -pinkish in color.
The color of A. perideraion eggs changed
from orange -pinkish to grayish pink and gray
during incubation.
Allen (1974) recordedthat A. chrysopterus
took 3 h to reach 8-cell stage , 5 h to
reach 32-cell stage and 10 h to enter the
blastula stage . At the blastulation stage ,
the cleavage process of A. perideraion was
recorded earlier than that of A. chrysopterus.
Falk-Petersen (2005) mentioned that the
blastula formed after hours or a couple of
days depended upon species or incubation
temperature. The early cleavage varies
depending on egg qual ity and fishes
(Kjorsvik et a/., 2003). Similarly, the embryo
of A. perideraion took 2 h, 2 h 25 min, 3 h, 3
h 30 min, 4 h 20 min, 16 h 45 min to reach
8, 16, 32, 64-blastomere, morula stage and
gastrula stage, respectively.
TAO
82
Yuan-Shing Ho, Che-Ming Chen, Wen-Vie Chenand Wen-Been Chang
Wilkerson (2001) reported that heart,
brain and spinal cord on Amphiprion embryo
formed on day 3 after deposited. The heart
beat was approximately 200 beats per
minute with the major blood circulation clearly
visible on day 4 for A. chrysopterus (Allen ,
1974). The embryo of A. perideraion on 78
h 30 min after fertilization, chromatoplasm
precipitated on eyes , and heartbeat rate
became 156 times/min, 30-somite appeared.
The appearance of the first pectoral fin on A.
chrysopterus embryo was recorded on the
fifth day (104 h) (Allen, 1974). Pectoral fin of
A. perideraion became functional while the
head and tail moved actively on day 5 (115 h
11 min) after insemination in this study.
Anemonefishes are known to have a
protective mucous coat that allows them to
contact the tentacles of their host anemone
without being stung. There are two conflicting
hypotheses as to the source and biochemical
properties of this mucous coating. One
hypothesis proposes that anemonefishes
acquire anemone substances from their
hosts during the behavioral process of
acclimation, which protect the fish from being
stung (Mar iscal , 1970b; Elliot et al., 1994) .
Anemonefishes are considered to use
anemone mucus as "chemical camouflage"
or "macromolecular mimicry" to avoid
recognition as "not-self' by the anemone,
and possible subsequent stinging. Another
hypothesis is that anemonefishes produce
their own protective mucus coat, which lacks
substances that elicit cnida (nematocyst
and spirocyst) discharge by their hosts
(Miyagawa, 1989). The sea anemonefishes benefit by receiving protection from
predators among the nematocyst-Iaden
tentacles of the sea anemone host, perhaps
by receiving some form of tactile stimulation,
by being less susceptible to various diseases
and by feeding on anemone tissue, prey ,
waste material and perhaps crustacean
symbionts (Mariscal , 1970a). A pyridinium
compound , amphikuemin , which induces
characteristic attracted swimming (toward
the chemical stimulus) in anemone fish
Amphipriun perideraion, has been isolated
from sea anemone Radianthus kuekenthali
(Konno et al., 1990). The mortality ratio of
A. perideraion larvae with change in color
and without any change in color (17 DPH)
in the present study has showed that the
development of protective systems in those
without any changes in color is incomplete.
The above mechanisms acting singly or
in conjunction, a critical period or catastrophic
mortality stage can be identified in the
reproductive cycle of marine fishes which
chronologically correlates very closely with
the period shortly after the yolk sac has been
fully absorbed. Periodic unavailability of food
may not only cause death through starvation
(Blaxter & Ehrlich , 1974), but can lead to
malnutrition (Ehrlich et al. , 1976), lower
growth (Theilacker & Watanabe, 1989),
reduced performance (Margulies , 1993)
and ultimately lower probabilities of survival
through increased predation (Neilson et al.,
1986; Rice et al., 1987; Miller et al., 1988).
Amphiprion spp. have a short larval period
(15-20 days) relative to other coral reef fishes
(Wellington & Victor, 1989). It is similar
to A. pe rideraion larvae (17 DPH) in the
present study. Recent studies have shown
that the body condition of newly settled fish
within and between recruitment episodes is
highly variable (McCorm ick & Molony, 1993;
Kerrigan , 1996), but at present , the causes
of this variability are unclear. The body
condition of newly settled fish may provide
an important insight into the environmental
conditions experienced by fish during their
larval phase (McCormick , 1998). A variety of
histological and morphological measures of
body condition have been used successfully
to discriminate between fish of different
feeding histories (Theilacker & Watanabe,
1989; Margulies, 1993; Theilacker et al.,
1996). However, the temporal response
of fish to food intake is likely to be species
specific, and more experimental studies are
required to facilitate interpretation in relation
to feeding history for species targeted for
research.
TAO
ACKNOWLEDGMENTS
We would like to thank Mr. Jin -Shan
Wang , Mr. Shi-Chang Lin (Eastern Marine
Biology Research Center) , and Mr. Shen-gi
Reproduction of PinkClownfish
Chen (aquaculture department of Pingtung
University of Science and Technology) for
their assisted in this study.
REFERENCES
Allen , G. R. (1974). The Anemonefishes. 2nd
ed . The class ification and biology. T.F.H.
Publication, Inc. USA. 352 p.
Arezo , M. J., L. Pereiro and N. Bero is (2005).
Early development in the annual fish
Cynolebias viarius. J. Fish BioI . 66: 13571370.
Blaxter, J. H. S. and K. F. Ehrlich (1974). Changes
in behaviour during starvation in herring and
plaice larvae. In The Early Life History of Fish
(Blaxter, J. H. S., ed.), pp. 575-588.
Doherty , P. J. (1988 ). Spatial and temporal
patterns in recruitment. In The Ecology of
Fishes on Coral Reefs (Sale, P., ed.), pp. 261294.
Ehrlich, K. F., J. H. S. Blaxter and R. Pemberton
(1976). Morphological and histological
changes during growth and starvation of
herring and plaice larvae. Mar. BioI. 35: 105118.
Elliott, J. K., R. N. Mariscal and K. H. Roux (1994).
Do anemonefishes use molecular mimicry
to avoid being stung by host anemones? J.
Exp. Mar. BioI. Ecol. 179: 99-113.
Falk-Petersen, I. B. (2005). Comparative organ
differentiation during ea rly life stages of
marine fish. Fish Shellfish Immun. 19: 397412.
Hoff, F. H . (1993) . Marine o rn a mental fish
culture. From discovery to commercialization.
European Aquaculture Society, Oostende
(Belgium). Spec. Publ. Eur. Aquacult. Soc. 19,
238.
Job , S., M. Arvedlund and M. Marnane (1997) .
Culture of coral reef fishes. Austasia Aquacult.
11(3): 56-59.
Kerrigan, B. A. (1996). Temporal patterns in size
and cond ition at settlement in two tropical
reef fishes (Pomacentridae: Pomacentrus
amboinensis and P. nagasakiensis). Mar.
Ecol. Pro. Ser. 135: 27-41.
Kjorsvik , E., K. Hoehne-Reitan and K. I. Reitan
(2003). Egg and larval qual ity criteria as
predictive measures for juvenile production in
turbot (Scophthalmus maximus L.). Aquacul-
83
ture 227: 9-20.
Kobayashi , M. and A. Hattori (2006) . Spacing
pattern and body size composition of the
protandrous anemonefish Amphiprion
frenatus inhabiting colonial host anemones.
Ichthyological Research 53(1): 1-6.
Konno, K., G. W. Qin, K. Nakanishi, M. Murata and
y. Naya (1990). Synthesis of amphikuemin
and analogs: A synomone that mediates
partner recognition between anemonefish and
sea anemone. Heterocycles, 30: 247-252.
Kovac , V. (2005). Early development of Zingel
streber. J. Fish BioI. 57: 1381-1403.
Leis, J. M. (1991). The pelagic stage of coral reef
fishes : the larval biology of coral reef fishes.
In The Ecology of Fishes on Coral Reefs
(Sale, P. F., ed.), pp. 183-230.
Margulies, D. (1993). Assessment of the nutritional condition of larval and early juvenile
tuna and Spanish mackerel (P isces ,
Scombridae) in the Panama Bight. Mar. BioI.
115: 317-330.
Mariscal , R . N. (1970a) . The nature of the
symbiosis between Indo-Pacific anemone
fishes and sea anemones. Mar. BioI. 6: 58-65.
Mariscal, R. N. (1970b). An experimental analysis
of the protection of Amphiprion xanthurus
and some other anemone fishs from sea
anemones. J. Exp . Mar. BioI. Ecol. , 4: 134149.
McCormick, M. I. (1998). Condition and growth of
reef fish at settlement: is it important? Aust.
J. Ecol. 23: 258-264.
McCormick, M. I. and B. W. Molony (1993) .
Quality of the reef fish Upeneus tragula
(Mullidae) at settlement: is size a good
indicator of condition? Mar. Ecol. Pro. Ser. 98:
45-54.
McLarney, B. (1985). Pioneers in saltwater
aquarium fish. Part I. Aquacult. Mag .
(November/December), 38-41.
McLarney, B. (1986 ). Pioneers in saltwate r
aquarium fish . Part II. Aquacult. Mag.
(January/February),31-33.
Meekan, M. G. and L. Fortier (1996). Selection for
fast growth during the larval life of Atlantic cod
Gadus morhua on the Scotia Shelf. Mar. Ecol.
Pro. Ser. 137: 25-37.
Miller, 1. J., L. B. Crowde r, J. A. Rice and E. A.
Marschall (1988). Larval size and recruitment
mechanisms in fishes: toward a conceptual
TAO
84
Yuan-Shing Ho, Che-Ming Chen, Wen-Vie Chen and Wen-Been Chang
framework. Can. J. Fish. Aquat. Sci. 45: 16571670.
Miyagawa, K. (1989). Experimental analysis of
the symbiosis between anemonefishes and
sea anemones . Ethology 80: 19-46.
Neilson, J. D., R. I. Perry, P. Valerio and K. G.
Waiwood (1986). Condition of Atlantic cod
Gadus morhua larvae after the transition
to exogenous feeding: morphometries ,
buoyancy and predator avoidance. Mar. Ecol.
Pro. Ser. 32: 229-235.
Rice , J . A ., L. B. Crowder and F. P. Binkowski
(1987). Evaluating potential sources of
mortality in larval bloater (Coregonus hoyt):
starvation and vulnerability to predation. Can.
J. Fish. Aquat. Sci. 44: 467-472.
Sale, P. F., J. A. Guy and W. J. Steel (1994).
Ecological structure of assemblages of coral
reef fishes on isolated patch reefs. Oecologia
98: 83-99.
Shuman , C. S., G. Hodgson and R. F. Ambrose
(2005). Population impacts of collecting sea
anemones and anemonefish for the marine
aquarium trade in the Philippines. Coral Reefs
24(4): 564-573.
Swanson, C . (1996) . Early development of
milkfish : effects of salinity on embryonic
and larval metabolism, yolk abso rption and
growth. J. Fish BioI. 48: 405-421.
Theilacker, G. H., K. M. Bailey, M. F. Canino and S.
M. Porter (1996). Variations in larval walleye
pollock feeding and condition: a synthesis.
Fisheries Oceanography 5 (Supplement 1),
112-123.
Theilacker, G . H. and Y. Watanabe (1989).
Midgut cell height defines nutritional status
of laboratory raised larval northern Anchovy
Engraulis mordax. Fish. Bull. 87: 457-469.
Wellington, G. M . and B . C . Victor (1989) .
Planktonic larval duration of one hundred
species of Pacific and Atlantic damselfishes
(Pomaemtridae). Mar. BioI. 101: 557-567.
Wilkerson , J. D. (1998). Clownfishes. A guide
to their captive care, breeding and natural
history. 1st ed. Microcosm Ltd. , Shelburne,
VT.240p.
Wilkerson , J. D. (2001). Clownfishes: A guide
to their captive care , breeding and natural
history. Microsom Ltd. USA. 240 p.
Young, F. A. (1996). The state of tropical marine
aquarium animal cultivation . Freshwat. Mar.
Aquar. 5: 126-129.
TAO
Repn叫uction
of PinkClownfish
85
粉紅海葵魚之生殖行為及育苗研究
何源興 1 . 陳哲 明 1 . 陳文義 1 . 張文炳口*
(2008 年2 月1 日 收件 ; 2008 年3 月15 日 接受 )
粉紅海 葵魚 ( A m p h ip ri o n perideraion) 屬 雀 綱 科 ( P o m a c e n t r i d a e ) 之 海 葵 魚 亞 科
(Amphiprioninae) 。 在247 日 的 研 究 期 間 共 計觀察 到 產 卵21 次 ; 如 同 時 觀察 到 種 魚 有 清潔
產卵床行 為 、雌魚腹部明顯 膨大 、生殖突起明 顯突出，則 可判定種魚即將產卵 。在 2 7
1 °C 、 鹽 度34
(n
±
1%。環 境條 件 下 ， 每 次 產 卵 聞 隔 約13.0
= 49) 。 產卵 時 間 在09 :15-14:05
±
3 .7 日 ， 孵 化 時 間 需7.2
.大部份 集 中在 0 9 : 0 0 - 1 1
:00
±
±
1 .2 日
.產卵行 為持 續約 1個多
小時 ;每次產卵 數 約 3 0 0 - 7 0 0顆，卵粒為橘紅色 、精圓 形 、分離之 沉性附著卵，長 徑 為
1.85 ± 0.20 (n
= 30) mm
.卵黃長徑為 1 . 3 0
± 0.13 mm
.內有許多油球，油 球徑 1 52
± 109
IJm 。 且 孵 化 日 之 晚 間 應保持 完全 黑 暗 狀 態 。 育苗 期 間 水 溫 ， 孵化 之仔 魚 全 長 為3.20-3 .80
mm · 並具驅光性 ， 隨著成長 ， 仔魚驅光性變 弱 。 孵化後1 -11 日 間 ， 投予輪 虫 ( Bra ch ion us
plicatilis)
，第 1 2 日起，即 可兼投撓足 類 ( C o p e p o d ) ，第 1 5 日以 後完全投予撓足 類 ， 孵化 後
第 59 日 ， 魚 苗之全 長為 1 6 . 62 -2 8 . 6 3
mm
'已 可以完全接 受人工粒狀飼料， 孵化後第 2 6 -3 0
日，魚苗體色斑紋大致和成魚相似 ， 孵化後第 2 2日之魚苗 已有領域及爭鬥行為 。體色已轉
變之仔魚 ，對海葵觸 手發 射 的 刺 細 胞始 開 始有免疫 的 功 能。
關鍵詞: 粉 紅 海 葵 魚， 生 殖 行 為， 胚胎 發 育， 育苗 。
TAO
1 水產試驗所東部海洋生物研究 中 心
2 國 立 東華大學海洋生物 多樣性及演化研究所學水產養殖學
3 國 立海洋生物 博物 館
* 通訊作者:
[email protected]