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Ovarian Development and Induced Oviposition of the Overwintering Swimming Crab Portunus Trituberculatus (Brachyura: Portunidae) Reared in the Laboratory

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Ovarian Development and Induced Oviposition of the Overwintering Swimming Crab Portunus Trituberculatus (Brachyura: Portunidae) Reared in the Laboratory Blackwell Science, LtdOxford, UKFISFisheries Science0919 92682004 Blackwell Science Asia Pty LtdDecember 2004706988995Original ArticleReproduction of female swimming crabK Hamasaki et al. FISHERIES SCIENCE 2004; 70: 988–995 Ovarian development and induced oviposition of the overwintering swimming crab Portunus trituberculatus (Brachyura: Portunidae) reared in the laboratory Katsuyuki HAMASAKI,1a* Hideyuki IMAI,2b Nobuhiko AKIYAMA3 AND Kyohei FUKUNAGA1c 1Tamano Station, National Center for Stock Enhancement, Fisheries Research Agency, Tamano, Okayama 706-0002 and 2Graduate school of Marine Science and Technology and 3Department of Fisheries, Faculty of Marine Science and Technology, Tokai University, Shimizu-orido, Shizuoka 426-8610, Japan ABSTRACT: Females of the swimming crab Portunus trituberculatus were cultured under natural temperature and high temperature (21∞C) conditions to examine ovarian development and oviposition from autumn (October) to the beginning of breeding season in the following spring (May). Ovaries developed because of vitellogenesis of oocytes from October to December and their developmental state did not change during the overwintering period from December to March. In spring, from late March to mid May, some ovaries reached prematuration and maturation stages and ovipositions began from mid–late April under natural temperature conditions. Females reared in tanks of high tem- perature regimes oviposited earlier than those reared in tanks of natural temperatures. The number of days to oviposition tended to decrease with advancement of the beginning times (early February to late April) of culture of females under high temperature regimes; induced ovipositions were achieved more easily with increasing photoperiod from c. 12 h in early February to c. 14 h in late April. There- fore, it is inferred that the temperature and photoperiod are important environmental factors controlling ovarian development and oviposition. KEY WORDS: ovarian development, oviposition, photoperiod, Portunus trituberculatus, Swimming crab, temperature. INTRODUCTION ovigerous females occur from April/May to August/September in the southern range of its dis- The swimming crab Portunus trituberculatus tribution in Japan.4–7 The females of P. tritubercula- (Miers) is distributed mainly on sandy and muddy tus are also known to oviposit several broods bottoms of bays and inlets from Hakodate, Japan, during a breeding season of the year.8 Larvae (first through Korea to China and Taiwan.1 In Japan, this zoeas) used for seed production were obtained species has been selected as a target species for from ovigerous females caught from the wild and stock enhancement programs through production from females oviposited in tanks.9 It has been sug- and release of juvenile crabs.2 Recently, the annual gested that larvae from the first broods have the production of juvenile crabs at hatcheries has advantage of easy rearing because their body size is reached c. 58 million.3 larger than that of larvae from successive broods.9 Ovaries of P. trituberculatus develop from Therefore, it is considered that induced oviposition autumn (October) to the following spring4–6 and before the breeding season is useful in obtaining larvae from the first brood for a longer period. For several crustaceans, studies have revealed *Corresponding author: Tel: 81-3-5463-0538. that temperature and photoperiod play an impor- Fax: 81-3-5463-0542. Email: [email protected] tant role in regulating ovarian development and aPresent address: Faculty of Marine Science, Tokyo University oviposition; for example, ovarian development is of Marine Science and Technology, Minato, Tokyo 108-8477, accelerated and oviposition is induced under long Japan. photoperiod and high temperature conditions.10–12 b Present address: Faculty of Science, University of the However, information on the reproductive biology Ryukyus, Nishihara, Okinawa 903-0213, Japan. cPresent address: Akkeshi Station, National Center for Stock of P. trituberculatus is very limited to reliably con- Enhancement, Fisheries Research Agency, Akkeshi, Hokkaido trol its ovarian development and oviposition for 088-1108, Japan. seed production; it is only reported briefly that ovi- Received 7 January 2004. Accepted 14 July 2004. positions were induced under high temperature Reproduction of female swimming crab FISHERIES SCIENCE 989 conditions from March–April before breeding were replaced in 70% ethanol solution after 24– season in a hatchery.13 72 h. Pieces of fixed ovary were dehydrated in The present study examined ovarian develop- ethanol, embedded in paraffin, sectioned 5-mm ment and oviposition of laboratory-reared P. tritu- thick, and stained with Mayer’s hematoxylin–eosin. berculatus from autumn (October) to the The ovarian developmental stage was expressed by beginning of breeding season in the following the most advanced stage of oocytes among 10 his- spring (May) under natural temperature condi- tological stages during oogenesis of P. tritubercula- tions. In addition, we analyzed the relationship tus reported by Imai et al.6 Five stages which were between the beginning times (early February to determined for the ovaries in this study were as fol- late April) of culture of females under high tem- lows: yolk granules are dyed with eosin in the yolk perature regimes and the number of days to granule stage; yolk granules fuse and become oviposition. larger peripherally to form yolk globules in the primary yolk globule stage; yolk globules increase in size and the nucleus begins to shrink and MATERIALS AND METHODS becomes basophilic in the secondary yolk globule stage; yolk globules partly fuse to one another and Ovarian development the nuclear membrane becomes ragged in the pre- maturation stage; and fusion of yolk globules Females which had copulated with males in the progresses and germinal vesicle breakdown occurs wild were obtained from natural habitats in the in the maturation stage. Maximum diameters of Seto Inland Sea of Japan: 43 from October–Decem- 200 oocytes, which were opaque as a result of vitel- ber 1991 and 10 in October 1992. Carapace width logenesis after yolk granule stage, from ovary lobes (CW) including lateral spines of females ranged fixed in formalin were measured using a Profile 148–235 mm (mean, 183 mm) and was larger than projector (V-12; Nikon Corp., Tokyo, Japan). the biological minimum (c. 120–130 mm in CW) of Although there were no histological differences this species.4,5 between oocytes from several parts of the ovary, Females were reared in our institute using 7-kL the collection site of subsamples for measure- (450 cm ¥ 183 cm ¥ 83 cm) tanks with bottoms ments of oocyte diameter were fixed at the top of covered with c. 10 cm of fine sand. Each female was the ovary. The relationship between modal values tagged with a numbered vinyl tape to facilitate in the frequency distributions of oocyte diameter individual identification. Tanks were located out- (x) and the GI values (y) was analyzed using a doors and covered with a blackout curtain (shade power function, y = axb. In the present study, the efficiency, 90%) that allowed natural light to enter. model to describe random error of y is exemplified Sand-filtered seawater was supplied using a flow- as a normal distribution Ny()ˆ,s22 yˆ , where the through system (15–24 L/min; salinity, 31–32 psu) standard deviation is proportional to the theoreti- and water temperatures were not regulated. cal value of yy()= ˆ by a certain factor s according to Females were fed with Manila clam Ruditapes Yamakawa and Matsuda.14 Parameters of the equa- philippinarum or frozen Krill Euphausia superba tion were estimated by the maximum likelihood every day. method detailed in the previous paper.15,16 In this study, gonad index (GI), histology, and diameter of oocytes were used to identify the ova- rian developmental state. One to five females were Induced oviposition sacrificed once a month and the total of 28 and nine surviving females were dissected from Octo- From 1986 to 1992, a total of 541 females (141– ber 1991 to May 1992 and in October and Decem- 252 mm CW) were collected in the Seto Inland Sea ber 1992, respectively. Rearing periods of females and reared in two to three 7-kL tanks in each year to the time of sacrifice ranged 27–169 days except and a total of 380 surviving females were used for for females that were sampled after 1–6 days from culture trials of induced oviposition (Table 1). collection in October and November. The CW Induced ovipositions were attempted by rearing (mm) of the specimens were measured with a slide the females under high temperature regimes caliper, then their ovaries were removed. Their (=heating culture) for a certain period during the wet weights (g) were measured after paper-blotting overwintering period in each year from 1987 to and GI was calculated using the formula, 1993, as shown in Table 2. Five to 47 females were GI = (Gonad weight) ¥ 107/CW3. The ovary was H- used for each trial and reared in one to three 4-kL shaped. Right and left lobes of ovaries were fixed in (450 cm ¥ 150 cm ¥ 50 cm) tanks with bottoms Bouin’s solution and 10% neutral formalin solu- covered with fine sand. These tanks had flow- tion, respectively. Ovaries fixed in Bouin’s solution through systems (8–14 L/min; salinity, 31–32 psu) 990 FISHERIES SCIENCE K Hamasaki et al. Table 1 Survival of females caught from the Seto Inland Sea until the beginning of culture trials for induced oviposition of Portunus trituberculatus in the laboratory Collection Females collected No. Date N CW† (mm) Survival (N/%) 19 Dec. 1986 31 213 (183–242) 19/61 2 11 Mar.-13 Apr. 1987 25 199 (155–238) 21/84 3 27 Nov.-30 Nov. 1987 74 209 (173–240) 43/58 4 20 Feb.-22 Apr. 1988 28 202 (158–233) 23/82 5 21 Nov.-24 Nov. 1988 53 218 (181–252) 43/81 6 24 Mar.-21 Apr. 1989 15 210 (182–235) 14/93 7 25 Nov.-13 Dec.
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