Aquacult. Sci.　63（2），207－212（2015）

Received 19 November 2014; Accepted 6 February 2015.1 Shimonoseki Marine Science Museum “Kaikyokan”, Shimonoseki Academy of Marine Science, Shimonoseki, Yamaguchi

750-0036, Japan.2 Department of Applied Aquabiology, National Fisheries University, Shimonoseki, Yamaguchi 759-6595, Japan.＊Corresponding author: Tel, (+81) 286-5111; Fax, (+81) 286-7435; Email, sakaih@fish-u.ac.jp (H. Sakai).

Note

Spawning and rearing of a porcupine puffer Cyclichthys orbicularis (Diodontidae, Tetraodontiformes) in captivity

Hiroyuki DOI1, Toshiaki ISHIBASHI

1 and Harumi SAKAI2,＊

Abstract: Cyclichthys orbicularis (Diodontidae, Tetraodontiformes) were spawned in captivity, the

larvae being reared for more than 60 days, in the Shimonoseki Marine Science Museum Aquarium.

Isolated epipelagic eggs, 2.2 mm in mean diameter, hatched two days after spawning. Hatched

larvae were 3.5 mm in total length, the head and trunk being covered by a “vesicular dermal

sac”. Fin rays were completed in 17 days after hatching at 7.6 mm TL, some rudimentary spines

appearing on the dorsum. Spines had become unmovable in 39 days after hatching at 20.8 mm TL.

Growth rates from hatching to 8 days and thereafter from 9 days were different, growth regression

equations between TL (y) and days after hatching (x) being y = 0.052x + 3.518 and y = 0.295x + 2.046,

respectively.

Key words: Porcupine puffer; Spawning; Development; Growth

Porcupine puffers of the family Diodontidae

include seven genera, 17 species and two sub-

species, variously distributed throughout tem-

perate to tropical seas worldwide (Froese and

Pauly 2014). Four genera, including seven

species, have been recorded from Japanese

waters (Aizawa and Doiuchi 2013). A few

reports on life history have indicated that iso-

lated epipelagic eggs are spawned, juveniles

and young fishes migrating to open pelagic

waters (Nishimura 1960; Fujita 1962; Leis 1978;

Sakamoto and Suzuki 1978; Fujita and Matsuura

2014), in contrast to tetraodontid puffers that

spawn demersal adhesive eggs (Fujita 1962;

Sakamoto and Suzuki 1978; Fujita and Matsuura

2014).

Two of the three known species of the genus

Cyclichthys have been recorded from Japan, Cy.

orbicularis (Bloch) and Cy. spilostylus (Leis and

Randall) (Aizawa and Doiuchi 2013; Froese and

Pauly 2014).

The present report details the first observa-

tions of spawning, eggs and larval development

for more than 60 days of Cy. orbicularis held

in the Shimonoseki Marine Science Museum

Aquarium. Although the observations were not

especially detailed, it was clear that this species

spawns isolated epipelagic eggs just as other

diodontids (Fujita and Matsuura 2014), the

larval and juvenile development being similar to

that of Diodon holacanthus Linnaeus, described

by Sakamoto and Suzuki (1978).

Materials and Methods

Wild-caught adults of Cy. orbicularis from

unknown locality, a female ca. 20 cm in stan-

dard length (SL) and a male ca. 15 cm SL,

208 H. Doi, T. Ishibashi and H. Sakai

were purchased from a fish dealer and main-

tained in a closed circulating-filtering exhibition

aquarium of 1.1 m3 (0.75×1.00×1.50 m) with

a large imitation coral rock on the coral sand

bottom, in the Shimonoseki Marine Science

Museum from August 2001. Water tempera-

ture was kept at 25℃ and subject to cyclic 12

h light and dark periods (Fig. 1A). Two indi-

viduals of Arothron nigropunctatus (Bloch and

Schneider) and each one of A. manilensis (Marion

de Procé), Tetrosomus concatenatus (Bloch),

Lactoria diaphana (Bloch and Schneider),

Lactophrys trigonus (Linnaeus), Chilomycterus

antillarum (Jordan and Rutter) and Ch. anten-

natus (Cuvier), all Tetraodontiformes, were

maintained in the same aquarium, being fed

three times weekly with processed giant tiger

prawns and Manila clams.

Although sexing of adult Cy. orbicularis is

usually very difficult, the belly of the female

became swollen several days before spawn-

ing. Both adults nestled close to each other on

the bottom the day before spawning (Fig. 1B).

Although the actual spawning behavior was

not observed, the female belly shrank after

spawning. Spawned eggs were found on five

separate occasions in May 21 (18 : 00), June 22

(9 : 00), August 27 (16 : 00), September 26 (9 : 00

and 11 : 30) and November 6 (12 : 00), 2004.

The eggs spawned in May and June were fer-

tilized but not so on the remaining occasions.

The rearing experiment was performed on the

second set of eggs spawned on 22 June 2004.

The rearing was continued more than 200 days,

but measurement and morphological observa-

tion were performed only from eggs to 49 days

after hatching.

When spawned eggs were found in the sur-

face layer of the aquarium, water samples

including the eggs were transferred to a 30 l

plastic tank and incubated at 25℃ with mild

airation.

Hatched larvae were sucked up with a glass

pipette and transferred to another 30 l plastic

tank, and reared at 25℃ without any special pH

control. The water was partly exchanged for

fresh water and excrement removed daily. The

tank was illuminated continuously over 24 h

daylight periods with two fluorescent lamps in

order to promote continuous feeding (Yoseda et

al. 2003).

Larvae, juveniles and young fish were fed

several times a day with Brachionus plicatilis

from the first 18 days, Artemia larvae from days

seven to 27, frozen Artemia adults from days

18 to 48, and processed krill, giant tiger prawns

and Manila clams from 48 days after hatching.

A

BFig. 1. The exhibition aquarium (1.1 m3; 0.75×1.00×1.50 m) in which the parental Cyclichthys orbicularis were

maintained and spawned (A); parents nestling to each

other on the bottom the day before spawning (B).

209Spawning of porcupine puffer

Results

Although the developmental stage was not

determined when found, the isolated epipelagic

eggs were spherical, 2.2 mm in mean diameter

(1.9-2.4 mm, n = 181), the embryo and primor-

dial eyes forming within one day of spawning

(Fig. 2A).

Hatching occurred two days after spawn-

ing. Hatched larvae (Fig. 2B, 2C) were 3.5 mm

in total length (TL), with the mouth and anus

closed. Membranous pectoral fins were obvious,

and the head and trunk covered by a “vesicu-

lar dermal sac” (Leis 1978) or “pliable shell”

(Sakamoto and Suzuki 1978). Melanophores

were scattered on the body, tail and belly, with

many xanthophores also on the body surface.

The eyes were not pigmented.

Nineteen hours after hatching, the eyes

had become pigmented and the mouth open

(3.5 mm TL, Fig. 2D). Two days after hatch-

ing, membranous dorsal and anal fins were

separated (3.5 mm TL, Fig. 2E). Five days after

hatching, soft rays were apparent in the pecto-

ral fins (3.7 mm TL, Fig. 2F), and seven days

later, soft rays appeared in the dorsal and anal

fins also (3.9 mm TL, no image).

Seventeen days after hatching, 12 dorsal, 10

anal and 21 pectoral fin rays were completed,

E

DC

A B

FFig. 2. Cyclichthys orbicularis egg of one day after spawning (2.2 mm in diameter) (A, live); hatched larvae of two

days after spawning (3.5 mm TL), (B, live) and (C); 19 h after hatching (3.5 mm TL) (D); two days after hatching

(3.5 mm TL) (E); five days after hatching (3.7 mm TL), (F, live). Scale bars indicate 1 mm.

210 H. Doi, T. Ishibashi and H. Sakai

with some rudimentary spines (lumps) appar-

ent on the dorsum (7.6 mm TL, Fig. 3A). By

twenty-three days after hatching, the rudi-

mentary spines had grown larger on the body

(9.8 mm TL, Fig. 3B), becoming unmovable 39

days after hatching (20.8 mm TL, Fig. 3C). After

spine development sequence was represented

by conditions at 52 days (about 20 mm TL, no

image) and 69 days (about 25 mm TL, Fig. 3D).

Growth of Cy. orbicularis from hatching to 49

days are shown in Table 1 and Fig. 4. Growth

rates from hatching to 8 days and thereafter

A B

C DFig. 3. Cyclichthys orbicularis young, 17 days after hatching (7.6 mm TL) (A); 23 days (9.8 mm TL) (B); 39 days

(20.8 mm TL) (C); 69 days (about 25 mm TL) (D, live).

Fig. 4. Growth of Cyclichthys orbicularis from hatching to

49 days. Growth rates from hatching to 8 days and thereaf-

ter from 9 days indicated by regression equations between

total length ( y) and days after hatching (x).

Table 1. Growth of Cyclichthys orbicularis from hatching to 49 days

Total Length (mm)

After hatching Mean 　SD＊ Range n

Just 3.5 0.1 3.2 - 3.7 48

19 hours 3.5 0.1 3.4 - 3.7 11

1 day 3.6 0.1 3.3 - 3.8 11

2 days 3.6 0.1 3.5 - 3.7 11

3 days 3.7 0.1 3.5 - 3.9 10

4 days 3.8 0.1 3.7 - 3.9 9

5 days 3.7 0.1 3.5 - 4.0 10

6 days 3.9 0.1 3.7 - 4.0 8

7 days 3.9 0.2 3.4 - 4.0 9

8 days 3.8 0.1 3.6 - 4.0 5

9 days 4.3 0.1 4.2 - 4.4 3

17 days 7.6 - - 1

23 days 9.8 - - 1

39 days 16.9 5.5 13.0, 20.8 2

43 days 14.6 2.3 11.9 - 20.0 10

44 days 13.4 2.0 11.1 - 16.6 7

45 days 15.6 2.4 12.7 - 19.3 7

49 days 20.0 - - 1＊Standard deviation.

211Spawning of porcupine puffer

from 9 days were clearly different as visually

shown in the growth lines in Fig. 4, regression

equations between TL (y) and days after hatch-

ing (x) being y = 0.052x + 3.518 (r = 0.663, P <

0.01, n = 132) and y = 0.295x + 2.046 (r = 0.809, P

< 0.01, n = 32), respectively.

Discussion

Cyclichthys orbicularis development was sim-

ilar to but faster in early and slower in later

stages than D. holacanthus (Sakamoto and

Suzuki 1978), with slightly larger stage sizes

(water temperature under 25℃ and 24.2-

27.9℃, respectively): eggs were 1.9-2.4 mm in

Cy. orbicularis (1.7-1.9 mm in D. holacanthus),

hatching occurred from 2 days after spawning,

hatched larvae being 3.2-3.7 mm TL (102 h

after spawning, 2.5-2.7 mm TL); eye pigmenta-

tion was 19 h after hatching at 3.4-3.7 mm TL

(24 h, 2.6-2.7 mm TL); fin rays were completed

and rudimentary spines appeared in 17 days at

7.6 mm TL (10 days, 4.9-5.9 mm TL); rudimen-

tary spines grew larger in 23 days at 9.8 mm

TL (13 days, 6.0-7.9 mm TL); spines were

completed in 39 days at 20.8 mm TL (16 days,

11.0 mm TL).

Cyclichthys orbicularis spawned isolated epipe-

lagic eggs, the larvae having a “vesicular dermal

sac” or “pliable shell”, as in other diodontids

(Fujita and Matsuura 2014). These character-

istics are shared among three tetraodontiform

families, viz. Diodontidae, Ostraciidae and

Molidae (Sakamoto and Suzuki 1978; Fujita and

Matsuura 2014), suggesting their close phyloge-

netic relationship (Sakamoto and Suzuki 1978).

Although the analysis based on larval character-

istics (Leis 1984) support such a relationship,

other tetraodontiform phylogenetic analyses

based on myology (Winterbottom 1974), spinal

cord morphology (Uehara et al. 2014), osteol-

ogy (Rosen 1984; Santini and Tyler 2003), and

DNA sequence (Yamanoue et al. 2008; Santini

et al. 2013) do not (Santini et al. 2013; Matsuura

2014), suggesting possible larval adaptation and

convergence.

Acknowledgments

We thank Tomo Akita, Eriko Akashi, Yuji

Inatsugu and Takayuki Sonoyama of the

Shimonoseki Marine Science Museum for tech-

nical help. Thanks also go to Dr. Graham Hardy

for help with English.

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メイタイシガキフグの水槽内での産卵と発育土井啓行・石橋敏章・酒井治己

　フグ目ハリセンボン科メイタイシガキフグ Cyclichthys orbicularis が下関市立しものせき水族館の水槽で産卵した。卵は分離浮遊性で平均直径は2.2 mm であり，2日でふ化した（25℃）。ふ化仔魚の全長は3.5 mm で，頭部および躯幹部は厚い外膜“vesicular dermal sac”で覆われていた。ふ化後17日，全長7.6 mm で各鰭条が完成し，背面に棘原基が出現した。ふ化後39日，全長20.8 mm で棘は不動となった。ふ化後 8日までと 9日以降では成長率が異なり，全長（ y ）とふ化後日数（x）の関係式はそれぞれ y = 0.052 x + 3.518 および y = 0.295 x + 2.046であった。