Japanese Journal of Ichthyology 魚 類 学 雑 誌 Vol. 36, No. 3 19 89 36 巻 3 号 1989 年

Larval Development, Growth and Age Determination in the Sharpnose Pufferfish Canthigaster valentini (Teleostei: Tetraodontidae)

Gregory J. Stroud, Barry Goldman and William. Gladstone

(Received August 23, 1985)

Abstract The eggs, early larvae and juveniles of the sharpnose pufferfish Canthigaster valentini are described, based on material collected in Great Barrier Reef waters. Eggs were obtained in the field by divers and reared in the laboratory. The eggs are spherical, strongly adhesive, 0.68- 0.72mm in diameter, possess a dense cluster of small oil droplets , and hatch around sunset 3 to 5 days after fertilization. Newly hatched larvae have a small yolk sac, pectoral fin folds, 17 myomeres (6 pre-anal, 11 post-anal) and measure 1.30-1.40mm in notochord (standard) length. The eggs of C. valentini differ from those of other tetraodontids in being much smaller and having a longer in- cubation time. The larvae can be distinguished from other tetraodontid larvae by pigmentation , myomere count and size at hatching. Growth is most rapid during the first day of larval life. Age determinations (based on otolith microstructure) of field collected juveniles , both pelagic and newly settled, indicate a pelagic phase of between 64 and 113 days for this species . This estimate appears consistent with the extended pelagic juvenile stages observed in other tetraodontiform fishes and could indicate that C. valentini can delay settlement for some time after becoming competent to settle at a minimum age of 64 days.

The tetraodontid subfamily Canthigasterinae conducted at the Lizard Island Research Station

(Tyler, 1980) is composed of a single genus Canth- in northern Great Barrier Reef of Australia.

igaster containing 24 species (Allen and Randall, Courting individuals of C. valentini were followed

1977; Lubbock and Allen, 1979). Despite the underwater by divers using SCUBA and the time

fact that members of the Canthigasterinae are and location of spawning noted. Eggs were

common over a wide geographical area, the egg collected attached to the algae in which they

and larval development has only been recently were laid (see below) a few minutes after spawn-

described in detail for one species: Canthigaster ing and transferred to the surface in plastic bags.

rivulata (Arai and Fujita, 1988). In the present They were then placed into buckets of fresh

report we describe the eggs, early larvae and seawater and aerated with a portable air pump juveniles of C. valentini (Bleeker) based on labor- until transported to the aquaria.

atory reared and field collected specimens. Data Pelagic juveniles were collected under night lights

on growth and ageing of larvae and juveniles, with a dip net and, except where noted otherwise,

obtained by examining daily increments on otoliths were immediately preserved in 80% ethanol. Set-

(Brothers et al., 1976; Struhsaker and Uchiyama, tled juveniles were collected from a series of small

1976) are also presented. C. valentini is one of artificial reefs using ichthyocides. These reefs were

the more common and widespread members of cleared of individuals weekly, therefore juveniles

the genus and is found on coral reefs of the tropical collected this way were never more than seven days

Indo-Pacific from shallow water to depths of at settled, assuming no inter-reef movement of set-

least 25 metres. A full desritption of the adult tled individuals.

is given by Allen and Randall (1977). Eggs were reared in 100 litre opaque, plastic,

circular tanks following the method of Houde and Materials and methods Ramsay (1971) and Houde (1973). Temperature during six rearing experiments ranged from 23.5

Field collections. Field work and rearing were to 27.1•Ž. Three days after hatching, larvae were

•\ 3 2 7•\ 魚類学雑誌 Japan. J. Ichthyol. 36 (3), 1989

fed on a diet of rotifer Brachionus plicatilis and (Taylor, 1967) in order to study ossified structures. wild size-sorted zooplankton. Natural illumina- Otolith Studies. Lapilli and sagittae were tion (under 70% shade cloth) was used at all times. removed from each larva by teasing them from A total of 93 larvae were sequentially removed and fresh or ethanol-preserved specimens with the preserved in 5% buffered seawater formalin and aid of fine stainless steel needles. The otoliths used for descriptive material. A further 16 were were transferred to clean microscope slides wetted taken along with the above series and preserved on with immersion oil. Generally, no further pre-

80% ethanol for otolith age-verification studies. paration was necessary-the otoliths were immedi- All specimens are housed in the larval fish collec- ately examined in immersion oil or, after drying, tion at the Australian Museum, Sydney. permanently mounted with the neutral mounting Terminology. Terminology of the developmen- medium -Pro-texx". Otoliths removed from tal stages follows that of Leis and Rennis (1983) juveniles were treated as above, but sometimes who consider the transition between larva and required etching for 1-2 minutes in a 1% solu- juvenile to coincide with the attainment of full tion of HC1 before being mounted (Struhsaker meristic characters and the loss of temporary and Uchiyama, 1976). specializations to pelagic life. The smallest growth increments of the mounted Meristics and morphometrics. All counts and otoliths were counted with a compound micro- measurements were made with a binocular dis- scope at magnification of 400-1,500•~after con- secting microscope and ocular micrometer. Ab- trast was maximized by a polarizing filter. In- breviations used in fin ray formulae follow those crements were counted from the nucleus to the of Leis and Rennis (1983). Illustrations were longest axis since they were most clearly discernible drawn with the aid of a camera lucida. in this region. Three consecutive counts were Definitions of morphometric measurements are made of the increments on each otolith: if the dif- as follows: standard length (SL) is the distance ference between these counts was <2 they were from the tip of the snout to the posterior tip of considered acceptable counts. In cases where the notochord (before hypural formation) or this difference was >2 the otolith was re-counted posterior margin of the hypurals; pre-anal length and either a series of counts accepted, or the data is measured from the tip of the snout along the discarded. Consistent (5% error) accepted counts midline to a vertical line through the posterior were obtained between the four otoliths of each edge of the anus; head length is the horizontal individual examined. Accepted counts were aver- distance from the tip of the snout to the upper aged over all four otoliths to give an estimate of edge of the gill opening; snout length is the hor- the age of an individual in days. This procedure izontal distance from the tip of the snout to the was unnecessary for laboratory reared larvae due anterior margin of the fleshy orbit; eye diameter to the ease and consistency in reading their otoliths. is the maximum horizontal distance measured Verification of the daily nature of otolith from anterior to posterior margin of the fleshy growth increments was obtained by comparing the orbit; pre-dorsal length is measured from the tip number of growth increments observed in the of the snout along the midline to a vertical line otoliths of laboratory reared larvae with the through the origin of the dorsal fin; body depth known chronological ages of these larvae. is the vertical distance between body margins (ex- clusive of fins) through the anterior margin of Results the pectoral fin base; body width is the transverse distance between body margins at the pectoral Spawning. At Lizard Island Canthigaster valen- fin base. tini spwans year round. This species has a male- Myomeres (pre-anal plus post-anal) were count- dominated social structure and undergoes pair ed on each larva when they could be distinguished. spawning between 0800 and 1530 hours. Spawn- Several larvae were stained with the vital protein ing generally takes place in coral rubble areas "Rose Bengal" to aid in counting myomeres where the eggs are laid on the botton attached to (Marcy, 1975). filamentous algae (Fig. 1A). Each of five females Osteology. Three juveniles (13.9-16.4 mm SL) followed laid a cluster of eggs numbering between were cleared with trypsin and stained with alizarin 15 and 876 (mean=314).

•\3 2 8•\ Stroud et al.: Development of Pufferfish

Fig. 1. Photomicrograph of Canthigaver valentini eggs. A: Eggs attached to filamentous algae. Each egg is approximately 0.70mm diameter. B: A single egg measuring 0.71mm diameter.

Description and incubation of eggs. Egg size 50 minutes.

ranged from 0.68 to 0.72mm diameter (mean Description of larvae and juveniles. At hatch-

0.70, n=49). The eggs were transparent, = spheri- ing, the larvae of C. valetitini possessed an ovoid cal and possessed a dense cluster of small oil head, a relatively deep body and measured 1.30- globules ranging from 0.01 to 0.05mm in diameter. 1.40mm in standard length. The small yolk sac The egg membrane consisted of nine radial layers contained numerous oil globules ranging from and has a strongly adhesive outer coating. Fine 0.01 to 0.05mm diameter along its dorsal and sediment sticking to this adhesive coating tended posterior margins ( Fig. 2A). The mouth was not

to give the egg an opaque, almost whitish ap- yet formed, the gut was straight and the anus

pearance (Fig. 1B). unopen. Each larva was enclosed in a dermal Incubation time of aquarium reared eggs, which sac which, except for small sections of the dorsal were kept attached to the algae on which they and ventral fin folds, was completely covered in were laid, varied inversely with temperature and numerous minute tubercles. The pectoral fin

ranged from 126.5 hours from spawning at 23.5•Ž folds were small, but present in newly hatched

to 81 hours at 27.1•Ž. However, this time was larvae. further constrained by the apparent requirement for By 24 hours posthatching at 23.5•Ž the yolk sac

hatching shortly after sunset. Hatching of all was considerably reduced and the mouth was six batches reared occurred between 1844 hours beginning to form. The anus was still unopen

(winter) and 2045 hours (summer) and was (Fig. 2B). There was no significant change in

always within one hour of nautical twilight (Nauti- morphology by 48 hours. By 72 hours posthatch-

cal Almanac, British Admiralty): range 14 to ing the yolk sac was almost completely absorbed,

―3 2 9― 魚類学雑誌 Japan. J. Ichthyol. 36 (3), 1989 the mouth was formed, the gut was coiled and ex- of 12 juveniles were D. 9, A. 9, P1. 16-17, C. 5+6. tended to about mid-body, and the anus was open These are within the range of adult values (Allen (Fig. 2C). Most of the smaller oil globules had and Randall, 1977). coalesced by this time and the larvae had com- Newly hatched larvae had unpigmented eyes menced feeding. Neither yolk nor oil globules although there was a band of pigment running were visible in larvae 6 days posthatching (Fig. from behind the posterior section of the eye to 2D, E). The gill opening had, by this time, closed the otic capsule (Fig. 2A). By 1.52mm SL (24 into a small pore. By 9 days posthatching the hours posthatching) this internal pigment had ex- nasal capsules were visible and the head and body tended further antero-dorsally (Fig. 2B). The had become slightly more robust (Fig. 2F). Noto- eyes began to develop pigment by the third day chord flexure had still not occurred. when the larvae were about 1.68mm SL and yolk Laboratory reared larvae grew from an average sac absorption was virtually complete (Fig. 2C). 1.35mm SL at hatching to an average of 1.82mm A large subdermal melanophore, situated approx- SL after nine days when the last of the survivors imately midway between the posterior margin of was taken for age studies. From the graph (Fig. the eye and upper end of the operculum, had 3) it can be seen that growth was most rapid during developed by the time larvae had reached 1.70 the first day of larval life. mm SL (6 days posthatching). A large stellate Morphometric growth and changes in relative external melanophore, just posterior to the base character size with growth is shown in Table 1 for of the pectoral fin fold, was also present at this several laboratory reared larvae and field collected stage of development (Fig. 2D). juveniles. It can be seen that there are marked Brain pigmentation appeared around 1.52mm differences in relative morphometric proportions SL (24 hours posthatching) in the form of a small with the change from larval to juvenile status. stellate subdermal melanophore situated postero- This change is least with the size of the eye which laterally on one, or both sides of the hindbrain gradually increases in relative size with larval (Fig. 2B). By 1.70mm SL (6 days posthatching) growth and reaches a maximum apparently about these small melanophores had been replaced by a the time of settlement, before regressing. Head single large stellate subdermal melanophore locat- length (and snout length), body depth, preanal ed on the posterior region of the hindbrain length and body width have all increased marked- (Fig. 2D, E). ly in juveniles as compared with the larvae, reflect- The yolk sac of a newly hatched larva exhibited ing the change towards robust body shape finally numerous small, scattered, stellate melanophores attained by these fishes. For the laboratory particularly along its dorsal margin where they reared larvae, body width increases to Day 3 then formed a line of heavy pigment (Fig. 2A). A decreases. This possibly indicates a loss of con- similar line, almost continuous with one of the dition of the larvae due to inadequate diet leading yolk sac, ran along the dorsal edge of the gut. One to death. to five small, stellate melanophores were present Myomeres were often obscured by the dermal on the myomeres situated above the gut region sac and/or heavy pigment, but in those larvae in (Fig. 2A) however, by 1.52mm SL (24 hours which they could be distinguished (1.30-1.64mm posthatching) these melanophores had disappeared SL, n=16) the total number of myomeres was (Fig. 2B). As development continued the gut re- always 17 (6 pre-anal plus 11 post-anal). The gion became heavily pigmented anteriorly, dorsally vertebral formula was given by Allen and Randall and dorsolaterally, but remained unpigmented (1977) and Tyler (1980) for C. valentini as 8+ ventrally (Fig. 2C, D, F). 9=17. Total myomeres thus agree 1:1 with ver- Tail pigmentation in newly hatched larvae con- tebral count (cf Miller et al., 1979), although the sisted of a series of melanophores which extended numbers of pre-anal myomeres and pre-caudal from around the level of the anus to five or six vertebrae differ. No fin rays were present in the myomeres caudad as well as from the dorsal to larvae up to age 9 days which had only fin folds. ventral body margins (Fig. 2A). By 1.52mm SL Fin rays most probably develop during late pre- (24 hours posthatching) the melanophores com- flexion and flexion stages as occurs in other tet- prising this series had become more numerous and raodontids (Leis and Rennis, 1983). Fin ray counts formed a distinct pigment band on the tail (Fig.

•\3 3 0•\ Stroud et al.: Development of Pufferfish

A

B

C

D

E

F

Fig. 2. Larval development of Canthigaster valentini. A, lateral view of newly hatched larva (1.32mm SL); B, 24 hours after hatching (1.52mm SL); C, 72 hours after hatching (1.68mm SL); D, 6 days after hatching (1.70mm SL); E, 6 days after hatching (1.70mm SL), dorsal yew; F, 9 days after hatching (1.82mm SL).

•\3 3 1•\ 魚類学雑誌 Japan. J. Ichthyol. 36 (3), 1989

Fig. 3. Comparison of standard length (mm) with age (days) for laboratory reared larvae at 23.0•Ž; mean

SE, n=31.

2B). This pigment band was present in all other vertebra, pterygiophores 2, 3, 4 and 5 were at- stages of larvae which were examined (Fig. 2C-F). tached to caudal vertebrae 2, 3, 4 and 4 respec- The pigmentation of juveniles was typical of that tively. The number and/or arrangement of dorsal of the adults albeit with fewer brown spots on the and anal pterygiophores thus differs somewhat head and body laterally. from that of Canthigaster rostrata described by Ossification is apparently completed before Tyler (1980). settlement as all adult structures were ossified in Age determination. Larvae were reared for up three newly settled juveniles (93-113 days old; to 7 days after hatching. Of the serveral counts 13.88-16.37mm SL) that had been cleared and of each of the four otoliths removed from each stained. The vertebral formula in each individual individual at days one, three, six and nine respec- was 8+9=17. The nine dorsal fin rays were tively, the number of rings counted agreed exactly supported by seven basal pterygiophores situated with the age in days except with one sagitta from between the neural spines of the seventh abdominal one of the nine day larvae which had only eight vertebra and the third caudal vertebra (i.e. between rings. There are thus no rings laid down during vertebra 7 and 11). The nine anal fin rays were the egg stage. supported through fibrous tissue by five basal Field collected newly settled juveniles ranged in pterygiophores. The dorsal edges of these pterygi- age between 64 and 113 days based on otolith ophores articulate by fibrous tissue with the ventral counts (Table 1). Actual age at settlement could, edges of the haemal spines of the caudal vertebrae: however, be up to seven days less than this as Pterygiophore 1 was attached to the first caudal settlement reefs were only visited once per week.

•\3 3 2•\ Stroud et al.: Development of Pufferfish

The one pelagic juvenile ageable gave an otolith raodontine genera Fugu, Sphoeroides, Tetraodon age of 105 days which, subtracting the five days (including subgenera Chelonodon, Monotreta and this individual was in an aquarium before extrac- Tetraodon) and Torquigener. The eggs of these tion of its otoliths, is consistent with the estimates species are typically spherical, 0.85-1.41mm in from settled individuals. No obvious "settle- diameter, contain a dense cluster of small oil ment" or transition marks of the type described by globules and are covered with an adhesive coating Victor (1982) and Brothers et al. (1983) were ob- (Welsh and Breder, 1922; Schreitmuller, 1930; served in the individuals examined. Munro, 1945; Uno, 1955; Fujita, 1962; Leis, 1984). These eggs are generally demersal and Discussion cemented to stones and rocks (Welsh and Breder, 1922; Schreitmuller, 1929, 1930; Randow, The literature on tetraodontid eggs and larvae 1934; Sterba, 1962; Honma et al., 1980; Richter, deals primarily with temperate species of the tet- 1982) or partly buried in the sand (Breder and

Table 1. Morphometric measurements (in percentage of the standard length) for larvae and juveniles of Canthigaster valentini. *Pelagic juveniles, the remainder being collected as settled individuals. **Damaged specimens. ?Otoliths unreadable.

•\3 3 3•\ 魚類 学雑誌 Japan. J. Ichthyol. 36 (3), 1989

Clark, 1947; Uno, 1955). The eggs of Canthigaster with a noticeably longer incubation time (126

valentini are also spherical, demersal, adhesive and hours at 23.5•Ž). possess a dense cluster of oil globules, but differ in In several species of tetraodontids the male guards being much smaller in diameter (0.68-0.72mm) the eggs until hatching occurs (Schreitmuller, 1930; and in being laid attached to clumps of filamentous Sterba, 1962; Richter, 1982). It has been reported algae (Gladstone, 1987a) rather than on rocks or that the males of Monotreta cutcutia also shelter in sand depressions. Richter (1982) reported that their hatched young in a sand depression and the freshwater puffer Carinotetraodon lorteti (named guard them for a further period in a manner not C. somphongsi in his article) spawned in algal unlike that of certain cichlids (Schreitmuller, loc. clumps, however his photographs show the eggs cit.). In contrast, no nest guarding or other form

to be scattered primarily over rocks. In Japan of parental care has been observed in C. valentini however, the marine pufferfish Fugu pardalis is (Gladstone, 1987a).

known to spawn in seagrass beds (Fujita, 1962). Larvae of C. valentini can be distinguished by

Arai and Fujita (1988) recently described spawning their smaller size at hatching (1.30-1.40 versus

in an aquarium by Canthigaster rivulata; unlike 1.60-3.20mm notochord length) and lower myo-

the former demersal spawning species a pair of mere count (17 versus 17-27) of the preflexion

C. rivulata released their gametes together above larvae of other tetraodontid fishes that have been

the substratum. described (cf. Welsh and Breder, 1922; Munro,

There is some additional information on the 1945; Fujita, 1956a, b, 1962, 1966; Fujita and

eggs of Canthigaster spp. Gladstone (1987b) re- Ueno, 1956; Shojima, 1957; Miller et al., 1977;

cently reported that the eggs, and larvae, of C. Leis and Rennis, 1982). The pigmentation,

valentini are unpalatable to several species of reef particularly the band of pigment extending from fish. A consequence of this is that parents do not the dorsal to ventral margins of the tail, is also

defend nests of fertilized eggs. Fujita (1962) very helpful in separating this species from others.

described the unfertilized eggs of C. rivulata as Few tetraodontid larvae have been collected in

transparent, spherical, 0.67-0.71mm in diameter plankton tows around Lizard Island (J. M. Leis,

with a mass of small oil droplets each measuring pers. comm.; Talbot, Goldman and Stroud, un-

0.017-0.035mm in diameter. Fujita (1962) also publ.) and of these only one species resembles C. noted that the egg membrane consisted of several valentini. The preflexation larvae of these two

radial layers. In a more recent paper Arai and species differ in length of the pigment band on the

Fujita (1988) described the fertilized eggs of this tail. In C. valentini the pigment band extends 5-8

species and counted 14 radial layers in the egg myomeres compared with only 3-5 myomeres in the

membrane. The eggs of C. rivulata thus appear other species which remains unidentified at the

to be very similar to those of C. valentini however, present time. Juvenile C. valentini are easily

C. valentini has a slightly larger egg diameter separable from juveniles of other Canthigaster

(0.68-0.72mm), a slightly greater range of oil species by colour pattern and meristics (Allen and droplet size (0.01-0.05mm diameter), and fewer Randall, 1977), which also distinguishes them from

layers in the egg membrane (nine), than the former other tetraodontid genera (Leis and Rennis, 1983).

species. The otolith age determinations of field collected

In the literature, the only other marine pufferfish juveniles indicate a pelagic phase of between 64 which produces eggs of similar size to C. valentini and 113 days for this species. The wide range

is the tropical inshore species Lagocephalus lunaris in values is probably not due to non-daily ring

spadiceus from Japan. Fujita (1966) described deposition, given the consistent results for the

their eggs as colourless, transparent, spherical, reared larvae. This range in age at settlement

0.61-0.70mm in diameter and possessing a cluster could indicate that C. valentini can delay settle-

of oil globules each measuring 0.009-0.07mm in ment for some time (as much as 49 days based on

diameter. He also mentioned that their eggs are our results) after becoming competent to settle at

demersal, but only faintly adhesive in nature. a minimum age of 64 days. Members of the

They had an incubation time of 76 hours at 21.0- tetraodontiform families Diodontidae, Monacan-

24.5•Ž. This contrasts with the eggs of C. valen- thidae and Balistidae also have extended pelagic

tini which are strongly adhesive, slightly larger and juvenile stages and are often of large size by the

•\3 3 4•\ Stroud et al.: Development of Pufferfish time they settle out from the plankton (Berry and Literature cited Vogele, 1961; Berry and Baldwin, 1966; Randall, Allen, G.R. and J.E. Randall. 1977. Review of the 1971; Randall and Klausewitz, 1973; Leis, 1978). sharpnose pufferfishes (Subfamily Canthigasterinae) There is also some evidence to suggest that at least of the Indo-Pacific. Rec. Austr. Mus., 30: 475-517. one ostraciontid species may be able to prolong Arai, H. and S. Fujita. 1988. Spawning behavior and the length of its pelagic juvenile stage (Moyer, early life history of the sharpnose puffer, Canth- pers. comm.; Thresher, 1985). Ambivalence in igaster rivulata, in the aquarium. Japan. J. Ichthyol., settling and consequent repeated re-entry into the 35 (2): 194-202. plankton (Fluchter, 1965; Marliave, 1977) may Berry, F.H. and W.J. Baldwin. 1966. Triggerfishes have also contributed to the wide range observed (Balistidae) of the Eastern Pacific. Proc. Calif. in newly settled individuals of C. valentini. Acad. Sci., 34: 429-474. Other reef fishes may have pelagic periods less Berry, F.H. and L.E. Vogele. 1961. Filefishes (Mon- variable and/or of shorter duration. Larvae of acanthidae) of Western North Atlantic. U.S. Fish Wildl. Serv., Fish Bull., 61: 61-109. Haemulon flavolineatum (Pomadasyidae) for in- Breder, C.M., Jr. and E. Clark. 1947. A contribution stance settle in 11-15 days (Brothers and to the visceral anatomy, development, and relation- McFarland, 1981) and Thalassoma bifasciatum ships of the Plectognathi. Bull. Amer. Mus. Nat. (Labridae) settles in 40-72 days (Victor, 1982). Hist., 88: 291-319. Brothers et al. (1983) examined the otoliths of 43 Brothers, E.B. and W. McFarland. 1981. Correla- species of newly settled reef fishes from One Tree tions between otolith microstructure, growth and life Island, Great Barrier Reef and found the otolith history transitions in newly recruited French grunts ages of individuals within a given species to vary (Haemulon flavolineatum). Rapp. P.-V. Reun. Cons. from one day in some species to 22 days in others. Perm. Int. Explor. Mer., 1978: 396-374. Unfortunately, only small numbers of individuals Brothers, E.B., C.P. Mathews and R. Lasker. 1976. Daily growth increments in otoliths from larval and were examined for the majority of species. Due adult fishes. U.S. Fish Wildl. Serv., Fish. Bull., to the limited available information on age at 74: 1-8. settlement in reef fishes at the present time we Brothers, E.B., D. McB. Williams and P.F. Sale. 1983. cannot say how typical Canthigaster valentini is of Length of larval life in twelve families of fishes at "One Tree Lagoon" reef fishes in its widely variable length of pelagic , Great Barrier Reef, Australia. stage. Mar. Biol., 76: 319-324. Fluchter, J. 1965. Versuche zur Brutaufazucht der Seezunge Solea solea in kleinen Aquarien. Hel- Acknowledgments golander Wiss. Meeresunters., 12: 395-403. (In This paper is a contribution from the Lizard German with English summary.) Island Research Station. The project was sup- Fujita, S. 1956a. On the development of the egg and prelarval stages of the puffer Fugu (Shosaifugu) ported by Commonwealth Research Grants to B. stictonotus (Temminck et Schlegel). Sci. Bull. Fac. Goldman and F.H. Talbot (MST 80/116) to F.H. Agr., Kyushu Univ., 15: 525-530. (In Japanese Talbot and B. Goldman (ARGC D179-16512). with English summary.) W. Gladstone was supported by a Commonwealth Fujita, S. 1956b. On the development of the egg and Postgraduate Research Award and Augmentative prelarval stages of the puffer Fugu (Shosaifugu) poeci- Research Awards from the Great Barrier Reef lonotus (Temminck et Schlegel). Sci. Bull. Fac. Marine Park Authority. The support of the Agr., Kyushu Univ., 15: 531-536. (In Japanese Australian Museum in Sydney and at Lizard with English summary.) Island is gratefully acknowledged. J. Leis as- Fujita, S. 1962. Studies on the life history and culture sisted in providing the pelagic juveniles, in helping of common pufferfishes in Japan. Nagasaki Pref. with taxonomic problems and in constructively Fish. Res. Stn. Rep., (2), 121pp., 40pls. (In Jap- anese.) criticising the manuscript. H. Sweatman helped Fujita, S. 1966. Egg development, larval stages and in the collection of newly settled juveniles and a rearing of the puffer Lagocephalus lunaris spadiceus colleague (who wishes to remain anonymous) as- (Richardson). Japan J. Ichthyol., 13: 162-168. sisted in setting up algal and Brachionus cultures. (In Japanese with English summary.) P. Schmitt helped in teachine one of us (GJS) Fujita, S. and M. Ueno. 1956. On the egg develop- otolith ageing techniques. Our thanks to all. ment and prelarval stages of Fugu (Torafugu) rubripes

•\3 3 5•\ 魚類学雑誌 Japan. J. Ichthyol. 36 (3), 1989

(Temminck et Schlegel). Sci. Bull. Fac. Agr., Copeia, 1971: 462-469. Kyushu Univ., 15: 519-524. (In Japanese with Randall, J.E. and W. Klausewitz. 1973. A review of English summary.) the triggerfish genus Melichthys, with description of a Gladstone, W. 1987a. The courtship and spawning new species from the Indian Ocean. Senckenb. behaviors of Canthigaster valentini (Tetraodontidae). Biol., 54: 57-69. Env. Biol. Fish., 20: 255-261. Randow, H. 1934. Tetraodon cutcutia and Tetra- Gladstone, W. 1987b. The eggs and larvae of the odon fluviatilis Hamilton-Buchanan. Wochenschr. sharpnose pufferfish Canthigaster valentini (Pisces: Aquar. Terrarienk., 31: 561-563. (In German.) Tetraodontidae) are unpalatable to other reef fishes. Richter, H.J. 1982. Spawning Somphongs' puffer Copeia, 1987: 227-230. Carinotetraodon somphongsi. Trop. Fish Hobbyist, Honma, Y., T. Ozawa and A. Chiba. 1980. Matura- 31: 8-25. tion and spawning behaviour of the puffer Fugu Schreitmuller, W. 1929. Tetraodon fluviatilus (Ham- niphobles, occurring on the coast of Sado Island in Buch) green ballfish. Aqu. Life, Baltimore, 13: the Sea of Japan (a preliminary report). Japan 23-26. J. Ichthyol., 27: 129-13. Schreitmuller, W. 1930. Kugelfishe. Das Aquarium, Houde, E.D. 1973. Some recent advances and unsolv- Berlin, Jan.: 12-16, Feb.: 20-26. (In German.) ed problems in the culture of marine fish larvae. Shojima, Y. 1957. On the development of eggs and Proc. World Maric. Soc., 3: 83-112. rearing of larvae of a puffer, Fugu (Higanfugu) pardalis Houde, E.D. and A.J. Ramsay. 1971. A culture (Temminck et Schlegel). Sci. Bull. Fac. Agr., system for marine fish larvae. Prog. Fish Cul., Kyushu Univ., 16: 125-126. (In Japanese with 33: 156-158. English summary.) Leis, J.M. 1978. Systematics and zoogeography of Sterba, G. 1962. Freshwater fishes of the world. the porcupine fishes (Diodon, Diodontidae, Tetra- Longacre Press, London, 878pp. odontiformes), with comments on egg and larval Struhsaker, P. and J.H. Uchiyama. 1976. Age and development. U.S. Fish Wildl. Serv., Fish. Bull., growth of the nehu, Stolephorus purpureus (Pisces: 76: 535-567. Engraulidae), from Hawaiian Islands as indicated Leis, J.M. 1984. Tetraodontioidei: Development. by daily growth increments of sagittae. U.S. Fish Pages 447-450 in H.G. Moser and W.J. Richards, Wildl. Serv., Fish. Bull., 74: 9-17. eds. Ontogeny and systematics of fishes. Special Taylor, W.R. 1967. An enzyme method of clearing Publication No.1, American Society of Ichthy- and staining small vertebrates. Proc. U.S. Natn. ologists and Herpetologists. Allen Press, Kansas, Mus., 122: 1-17. ix+760pp. Thresher, R.E. 1985. Reproduction of reef fishes. Leis, J.M. and D.S. Rennis. 1983. The larvae of T.F.H. Publ., Neptune City, New Jersey, 399pp. Indo-Pacific coral reef fishes. Univ. N.S.W. Press, Tyler, J.C. 1980. Osteology, phylogeny and higher Sydney, 269pp. classification of fishes of the Order Plectognathi Lubbock, R. and G.R. Allen. 1979. Canthigaster (Tetraodontiformes). NOAA Tech. Rep., Circ. 434, leoparda a new sharpnose pufferfish (Teleostei: 420pp. Tetraodontidae) from the central Indo-Pacific. Rev. Uno, Y. 1955. Spawning habit and early development Fr. Aquariol., 6: 87-90. of a puffer Fugu (Torafugu) niphobles (Jordan and Marcy, B. 1975. Mechanics of handling Ichthy- Schneider). J. Tokyo Univ. Fish., 41: 169-183. oplankton. Pages 11-12 in Ichthyoplankton. Re- Victor, B.C. 1982. Daily otolith increments and port of the CICAR Ichthyoplankton Workshop. recruitment in two coral reef wrasses, Thalassoma UNESCO Technical Papers in Marine Science No. asciatum and Halichoeres bivittatus. Mar.bif Biol., 20, Div. of Marine Sciences, UNESCO, Paris, 46pp. 71: 303-308. Marliave, J.B. 1977. Substratum preferences of set- Welsh, W.W. and C.M. Breder Jr. 1922. A con- tling larvae of marine fishes reared in the laboratory. tribution to the life history of the puffer Spheroides J. Exp. Mar. Biol. Ecol., 27: 47-60. maculatus (Schneider). Zoologica (N.Y.), 2: 261- Miller, J.M., W. Watson and J.M. Leis. 1979. An 276. atlas of common nearshore marine fish larvae of the Hawaiian Islands. Sea Grant Misc. Rep. UNIHI- (GJS: Centre for Environmental and Urban Studies, Seagrant-MR-80-02, 127pp. Macquarie University, North Ryde, NSW 2109, Aus- Munro, I.S.R. 1945. Postlarval stages of Australian tralia; BG: Lizard Island Research Station, PMB 37, fishes. No.1. Mem. Qld. Mus., 12: 136-153. Cairns, Qld. 4780, Australia; BG, present address: Randall, J.E. 1971. The nominal triggerfishes (Balis- Australian Museum, P.O. Box A285, Sydney South, tidae) Pachynathus nycteris and Oncobalistes eryth- NSW 2000, Australia; WG: Centre for Environ- ropterus, junior synonyms of Melichthys vidua. mental and Urban Studies, and School of Biological

•\3 3 6•\ Stroud et al.: Development of Pufferfish

Sciences, Macquarie University, North Ryde, NSW 受 精 後3-5日 の 日没 時 に ふ 化 す る.ふ 化 仔 魚 は 小 さ な 2109, Australia; WG, present address: NSW Depart- 卵 黄 嚢,胸 鰭 を そ な え,筋 肉 節 数17(6+11)で 脊 索 長 ment of Agriculture, Fisheries Research Institute, PO (SL)1.30-1.40mmで あ る.シ マ キ ンチ ャ ク フ グ の 卵 Box 21, Cronulla, NSW 2230, Australia) は 卵 径 が ず っ と 小 さ い こ と と ふ 化 所 要 時 間 が 長 い こ と で 他 の フ グ 類 の 卵 と 異 な る.本 種 の 仔 魚 は 色 素 細 胞 の 分 布, 筋 肉 節 数 及 び ふ 化 時 の 大 き さ で 他 の フ グ 類 仔 魚 と 識 別 で シ マ キ ン チ ャ ク フ グ の 仔 魚 の 発 生,成 長 と 日 令 決 定 き る.成 長 は 仔 魚 第1日 目 が 最 も急 速 で あ る.浮 游 生 活 Gregory J. Stroud・Barry Goldman・ 期 な ら び に 新 た に 定 着 し た 野 外 採 集 稚 魚 の 日令 決 定(耳 William Gladstone 石 の 微 細 構 造 に も と つ く)結 果 は,本 種 の 浮 游 生 活 期 が Great Barrier Reefで 採 集 した 材 料 に 基 づ い て,シ 64日 な い し113日 で あ る こ と を示 し て い る.こ の 概 算 マ キ ン チ ャ ク フ グCanthigaster valentinの 卵,初 期 仔 は 他 の フ グ 目魚 類 で 認 め ら れ て い る 延 長 浮 游 稚 魚 期 と も 魚 及 び 稚 魚 に つ い て 述 べ る.卵 は 野 外 で 潜 水 夫 に よ っ て 矛 盾 し な い よ うで あ り,シ マ キ ン チ ャ ク フ グ が64日 の 採 集 さ れ,実 験 室 で 飼 育 さ れ た.本 種 の 卵 は 球 形,強 粘 最 小 日令 で 定 着 能 力 を 得 た 後 は,あ る 期 間 定 着 を 後 ら す 着 性 で 卵 径0.68-0.72mm,小 油 球 の 濃 密 な 塊 を も ち, こ と が で き る こ と を 示 し て い る も の で あ ろ う.

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