330 Reproduction and metabolism of (Batillus) cornutus in Chiba, Japan.

SIONE VAILALA MATOTO,1 TOSHIATU SHIMIZU,2 HISANORI MITA,2 KOTARO TSUCHIYA,1 AND SUSUMU SEGAWA.1 1 Tokyo University of Fisheries , Tokyo 108 8477, Japan, 2 Chiba Prefectural Fisheries Research Center, Chiba 295-0024, Japan.

SUMMARY The reproduction and oxygen consumption of Turbo (Batillus) cornutus were studied in specimens collected monthly between March, 1999 and April, 2000 in Chikura, Chiba Prefecture, Japan. Gonadal histology, gonadal index and oocyte diameter were used to determine the reproductive cycles and spawning period for both sexes. Oxygen consumption and ammonia excretion were measured to estimate metabolism. The reproductive cycle was annual, with the peak activity in the late summer. Oxygen consumption and ammonia excretion increased linearly with increasing soft body weight and showed an annual cycle in connecting with gonadal maturation cycle.

KEYWORDS: reproductive cycle, oxygen consumption, gonad maturation, spawning, oocyte,

INTRODUCTION Gonad index (GI): The visceral coil of each individual was fixed and preserved in 10% formalin The Japanese turban top shell, Turbo (Batillus) seawater. GI was determined by measuring the cornutus, represents one of the most common proportion of the area occupied by the gonad to total of edible gastropods in Japan1).T. (B) cornutus inhabits area of the visceral coil section1,2,4). The GI was shallow rocky coast and it is an important coastal calculated using the formula: resource in Japan1,2).Its geographical distribution is restricted only to Japanese coastal waters (except northeastern Honshu and northern and eastern Each preserved visceral coil of the specimens was Hokkaido) and the southern part of Korean Peninsula3). sectioned, following the method established for T Although this is an important species as fisheries cornutus1). The section just behind the stomachal resources and there is need for information on fisheries caecum was used for GI measurement. biology, few comprehensive works have been made in Gonad histology: The gonad-digestive gland reproductive biology and physiology of this species 1,2,4) complex was dehydrated, embedded in paraffin, Monitoring the maturation process is important for the sectioned at 5-8 ƒÊm and stain with Harris Hematoxylin management and sustainable utilization of the and Eosin. resources. The present study was carried out to Based on the histological observation of determine the reproductive cycle of T. cornutus from Chikura, Chiba Prefecture. Oxygen consumption and gametes, gonad maturation process was categorized according to the criteria for T. (B) cornutusl,2,4). ammonia excretion of T. cornutus were measured as an Diameters of the oocytes were measured with indicator of the metabolism and activity in relation micrometer for 150 oocytes or ova with visible nucleus with maturation. in each individual from 4-7 female histological specimens. MATERIALS AND METHODS Oxygen consumption and ammonia excretion: Specimens of live T. cornutus selected from monthly A total number of 525 T. cornutus with shell height samples were used for measurement of oxygen of 63-111mm were collected monthly from March, 1999 to April, 2000 from Chikura, Chiba Prefecture. consumption and ammonia excretion. The total wet Sexes of T. cornutus were determined based on the soft body weight (W) ranged from 17.1 to 72.4g. colour of gonads5).The whole specimens of female All the subjected to the experiment were in and male collected every month were subjected to a post-digestive condition. They were starved at least for chi square goodness-of-fit test to test the null 2 days and acclimated into experimental condition in hypothesis on the sex ratio is 1:1. which average temperature and salinity were 20•Ž and 35 ppt, respectively. Following the acclimation for 4-6 hrs in the respiratory chamber under flow through 331

condition, dissolved oxygen concentration was Gonad Maturation and Oocyte Size Frequency measured by the Winkler titration method66 and ammonia contents by Indophenol method). Five stages of gonad development were identified based on the histological analyses: stage 1, early RESULTS active; stage 2, late active; stage 3, ripe; stage 4, partly spawn; and stage 5, spent. Gonad development Sex Ratio in stage 3 appear from April to October, stage 4 July to December, and stage 5 from September to January.

Sex ratio (male to female) ranged from 0.7 to 1.3 (1.1 Stage 1 was present almost throughout the year 0.2). Since the Chi-square test value did not except July and August, indicating that

significantly different (ƒÔ2 p>0.05) from expected male gametogenesis for next season started from to female sex ratio of 1:1, the population of T. cornutus September. Stage 2 for the next season appeared from from Chiba Prefecture consists of equal number of January and developed into stage 3 from April. male and female all through the year. Spawning started from July and end in October. Oocytes diameter T. cornutus ranged from <10 to Gonad Index (GI) <260 ƒÊm (Fig.2). Large oocytes (>140 ƒÊm)

predominated during a period from May to August, GI in both sexes rose from March 1999 with a peak while small oocytes (<80 ƒÊm) predominated from (around66%) in August and dropped to around 40% October to April. Although large number of small in September(Fig.1). In October, GI declined further oocytes and large oocytes (160ƒÊm< to <200ƒÊm) were

to about 20%, dropped to the lowest (<20%) in present throughout the year, no oocyte larger than Novemberand gradually increased until March 2000. 200ƒÊm was observed in October and November. A

In April 2000, GI was slightly lower than March. decrease in the number of bigger oocytes (>160ƒÊm) Duringthe increasingphase in GI of both sexes from started from September onward, with increase in

Marchto August 1999, GI slightly dropped from June number of small oocytes (<100ƒÊm) indicated that to July before reaching the peak in August. Seasonal changesof GI in both sexes showed similar patterns peak spawning period occurred during a period from late August to September. Oogonia and small oocytes (p>0.05),which indicates a strong synchronization in reproductivedevelopment between male and female (<40ƒÊm) were present throughout the year with throughoutthe year. progressive increase in number starting from September to April. This indicates that gametogenesis in T. cornutus is initiated on and after spawning. Size frequency distribution of oocyte (Fig.2) indicated seasonal changes of gonad maturation.

Oxygen consumption and ammonia excretion

R/W ranged from 47.7 to 82.3 (ml O2/kg W/hr) for T. cornutus 17.1-72.4 g. Figure 3(A) shows the monthly change in mean R/W (ml O2/g W/hr) of T. cornutus. Monthly fluctuation in R/W of male and female

showed similar patterns throughout the year. High R/W was recorded between August to October, while R/W in other months was relatively lower and constant. During the high R/W period, GI changed Fig. 1. Seasonal changes of gonad index and seawater with a peak spawning activity. At this period, the temperature. Error bar indicates standard deviation . seawater temperature was level off and remain around 25•Ž. Therefore, this change of R/W with high value The GI increased with rise in seawater temperature during Aug-Oct period may be resulted from the to a peak in August 1999 , except a slight drop in July effect of maturation and spawning. However, this (Fig.1). When the seawater temperature was level off (above 25•Ž) between August and October, GI phenomenon needs to be further elucidated by more detailed research in future. drastically changed from the peak of more than 60% in

August to a low value of around 20% in October. 332

Fig. 2. Percentage frequency distribution (abscissa) for different oocytes/ova size class (ordinate).

in October. When the spawning season starts in July, Monthly changes in E/W for both sexes is showed in Fig.3(B). E/W in this study ranged from 0.05-0.43 ON ratio was at its lowest and peak O/N ratio coincided with end of spawning season. (ƒÊg-atNH4+-N/gW/hr) at the same condition and ranged of sizes used in R/W. In June and July DISCUSSION 1999, and April 2000, E/W was relatively higher compare to other months during this study. The high Gonad index has been used as a good indicator to E/W value in April, June, July coincided with the estimate the reproductive maturation of gastropods period of progress in gonad maturation while they 1,2,4,8,9) utilised protein in high proportion in metabolism and Combining the information of GI with gonad used lipid for vitellogenesis. E/W for both sexes developmental stages composition and oocyte size fluctuated similarly but different pattern from R/W. frequency distribution, the population of T. cornutus in Chiba Prefecture was estimated to have a single major spawning period a year. This spawning period

peaks in late August through September, which agrees with the results of previous study in Chiba1). Spawning activity of wild population of T. cornutus was from June to November at temperatures ranging from 20 to 25•Ž 1,4,10,11) Based on the histological examination, gametogenesis in T. cornutus was classified into five stages as in previous studies on the same species1,4,12) and generally the same process for gastropods8,9,13) Two gametogenic periods occurred during the reproductive cycle of T. (B) cornutus. The first period occurred during the spring and summer and correlated well with the period of large increase in gonad size (early active, late active and ripe stages); the second period occurred immediately after spawning (partially spawn and spent) and was not reflected as a large change in gonad index. The R on individual basis in the present study

increases with increase in W as described by R=aW, where b range from 0.54 to 1.2 with a mean value of 0.9. The b value reported for 15 different bivalves range from 0.6 to 0.93 with their mean of 0.7214) In Fig. 3. Monthly changes in mean value of R/W (A) and Eucidaris tribuloides, b value was 0.70 during winter E/W (B) at 20•Ž. Error bar indicates the standard (20•Ž) and 0.65 in summer (30•Ž)15). The mean b deviation. value in this study indicates that metabolism of TT

cornutus is almost proportional to body weight. The O/N ratio was scattered widely during the The R/W (ml 02/g W/hr) in the present study reproductive cycle of T. (B) cornutus in this study. varied with season during the reproductive cycle and The mean O/N ratio ranged from 24.9 in July to 54.9 ranged from 0.80 to 1.37 mlO2/kg W/min at average

temperature of 20•Ž. T. cornutus studied by 333

temperature of 1620•Ž. T. cornutus studied by ) by the dimorphic characteristics of the pappilae of the right kidney opening. Venus (Jap. Jour. Malac.). Yamamoto et al.16) reported values of 1.26 and 1.46 1994; 53: (3) 229-239. mlO2/kgW/min at 23•Ž and 28•Ž, respectively. 6. Strickland JDH, Parsons TR. A practical handbook of Therefore, the values of R/W in this study are seawater analysis. Bulletin of Fisheries Research Board of comparable and within those reported for the same Canada., 1968; Vol. 167. specie 16) 7. Solorzano L. Determination of ammonia in natural water As the gonad matured towards spawning period, by phenol hypochlorite method. Limnol. Oceanogr., 1969;

R/W becomes higher during the period of peak 14, 799-801. spawning activity. Mytilus edulis increased R/W in 8. Young JF, DeMartini JD. The reproductive cycle, gonadal histology, and gametogenesis of red abalone, Haliotis winter in conjunction with gametogenesis14,17), but no rufescens (Swainson). California Fish and Game. 1970; 56: effect of gonadal condition and sex on R/W for (4) 298-309. Eucidaris tribuloides15) 9. Hahn K. The reproductive cycle of tropical top shell, O/N ratio is used as good indicator of metabolite, Trochus niloticus, in French Polynesia. Invertebr. Reprod. especially that of protein 21). Although the O/N ratio and Devlop., 1993; 24: (2) 143-156. calculated in this study was varied widely, the ratio 10. Ai T. Spawning and early development of the topshell, was lower in June, July and April which indicates the Turbo cornutus Solander-‡U. Induction of spawning and use of protein at higher rate as metabolite during this larval development. Bull. Jap. Soc. Sci. Fish., 1965: 31: (2), 105-111. period. During the spawning and late active stages 11. Rho S. Experimentation of top shell aquaculture. 1. and other season, the relatively high O/N ratio was Spawning and development of top shell, Turbo cornutus retained, and this indicate that the main metabolite Solander. Korean Fisheries Bulletin, 1976; 9 (1): 43-55. was lipid rather than protein. The pattern O/N ratio in 12. Lee JH. Gametogenesis and reproductive cycle of topshell this study is comparable the one has been described Turbo cornutus Solandar. Bull. Korean Fish Soc., 1980; 13: for bay scallop, Argopecten irradians concentricus's18). (4), 125-143. 13. Webber HH, Giese AC. Reproductive cycle and

ACKNOWLEDMENTS gametogenesis in black abalone Haliotis cracheroidii

(Gasdropoda: Prosobranchiata), Marine Biology 1969; 4: The present research was partially supported by the 152-159. Ministry of Education, Science, and Culture of Japan 14. Bayne BL. Thompson RJ, Widdows J, Physiology: I. In: B.L.Bayne (ed.) Marine mussels, their ecology and as a scholarship grant. We also appreciate to physiology. Cambridge University Press, London, 1976; Associate Professor Hiroshi Yamakawa for his 121-206. helpful advice in part of this research. 15. McPherson BF. Feeding and oxygen uptake of

tropical sea urchin Eucidaris tribuloides (L). Biol.

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