Four experimental diets were used to determine the effect of additional arginine in the high defatted soybean meal (DSM) diet on the protein retention and the growth of giant gouramy, Osphronemus gouramy Lac. Diet A contained 50% of the protein from brown fish meal (BFM) and 50% from DSM; diet B contained 50% BFM and 50% DSM plus 0.70% arginine; diet C contained 25% BFM and 75% DSM and diet D contained 25% BFM and 75% DSM plus 0.60% arginine. Fish, with mean bodyweight of 21.6 ± 1.13 g, were fed on the experimental diets for 56 days. The DSM diet (diet C) produced lower protein retention (PR), lipid retention (LR), feed efficiency (FE) and daily growth rate (DGR) than diet A (50% BFM + 50% DSM). The addition of arginine in diet D (75% DSM in dietary protein) can improve the DSM quality, which was shown by producing the same PR, LR, FE, and DGR as in diet A. These results indicated that arginine is one of the important essential amino acids for the high defatted soybean meal diet of giant gouramy.

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This study provides information on reproduction and food habits of Galeus eastmani and G. nipponensis in the east and west of Suruga Bay. A total of 731 G. eastmani and 754 G. nipponensis were sampled by commercial bottom trawlers in 1981-1989 and 1992-1996. Size at sexual maturity of both species of Galeus differed between the former and latter period. In G. eastmani, maturity size of females from the east tended to be smaller than those from the west. Both species laid large yolky ova the year round. Both species preyed on a wide variety of marine organisms, mainly Osteichthyes, Cephalopoda and Crustacea. Foods of G. eastmani differed between the east and west by season. Foods in the young stage of G. nipponensis differed from those of immature and mature by season. Body size of the young G. nipponensis was similar for mature G. eastmani, however the young fed on different organisms from G. eastmani in the east. Both species were voracious and opportunistic feeders.

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Growth curves of the psychrotrophic Vibrio isolates showed an increased slope throughout growth (in Type I strains) or a decreased slope after the stationary phase of growth (in Type II strains). Type I and II strains had optimum growth temperatures of 15 to 25°C and 20 to 30°C, respectively. Adenylate levels in Type I strains were markedly high for AMP, moderate for ADP, and low for ATP, and the major part of adenosine phosphates in Type II strains was occupied by AMP during the growth phase with all of them disappearing during the death phase, regardless of the growth temperatures. In Type I strains the concentrations of AMP, ADP, and ATP increased with growth, and in Type II strains the AMP concentration rose quickly, reached a maximum, and fell abruptly to the base during growth, independent of the growth temperatures. The values of energy charge during growth showed as 0.38 or below in both Type strains during growth at optimum temperatures or below. These results differed from those in Escherichia coli. Moreover, it was suggested that the strains of the same Type comprise the same ecological groups. The results indicate that the two Type strains are composed of different ecotypes, and the low energy charge and the dominance of AMP are characteristic of the psychrotrophic Vibrio spp.

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Otolith increments of bluefin tuna Thunnus thynnus were examined to estimate the periodicity of an increment formation and the timing of the first increment formation using larvae and juveniles with known age reared in three laboratories. The increment was formed daily up to 71 days after fertilization. The first increment was formed on the fifth day after fertilization, and around the fourth day after hatching which corresponded to the onset of feeding. Slower growing fish had otoliths with less distinct increments. Since wild captured larvae had otoliths with more distinct increments, fish in the wild either grow faster than that in a laboratory or only faster grown fish survived in the wild.

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Seven natural populations of climbing perch Anabas testudineus collected from the central, eastern, and peninsular areas in Thailand were assayed by allozymic analysis to examine genetic relationships among geographic populations. Twelve out of 18 enzyme loci were polymorphic. Significant differences in allelic frequencies were observed in all pairs of populations at one or more loci, and six loci showed the regional characteristic allelic compositions. Genetic distance between populations was higher between areas, in comparison with those within an area, being significant between the peninsular and the other populations. According to a UPGMA tree based on a genetic distance matrix, the seven populations fell into three major clusters in line with the area divisions. These groupings indicate that genetic relationships among climbing perch populations are much the same as for other freshwater fishes, and would depend on geographic features such as the river systems in Thailand.

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An experiment was conducted to determine the impact of exogenous thyroid hormone on metamorphic events and morphogenesis of larval Japanese flounder Paralichthys olivaceus. Six groups of 600 larvae each were immersed in 10 nmol thyroxine-sodium salt (T₄) during different developmental stages: A and B, C and D, E and F, C to F, and A to F. T₄ treatment during stages E and F resulted in premature metamorphosis including not only advanced settlement but also advanced eye migration of the juvenile fish. T₄ treatments also resulted in rapid absorption of the dorsal fin rays during stages C to F. As a result, metamorphosis to the bottom juvenile was induced by 10 nmol T₄. It was also found that the rate of metamorphosis was proportional to the T₄ immersion period. These results confirm that T₄ plays a major role in metamorphic events and morphogenesis during the early life stages of flounder. Thus, this study suggests that sensitivity to T₄ depends on the larval organ and the larval developmental stage.

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A mathematical model of the midwater trawl system is presented to aid in understanding the dynamic behavior. The equations of motions for the trawl system have been obtained using Lagrange's equation with respect to variables of the displacement of the trawler, the length of the warp connecting the trawler and otter board, the angle of the warp to the horizon and the angle of the hand rope to the horizon. The parameters included in the described model have been estimated using the data of the trawl system movements that were obtained from the midwater trawl towing by the training ship Kaya of Pukyong National University. The movements of the trawl system, calculated using the mathematical model, closely corresponded to the motions of the actual trawl system obtained through the field experiments. The model effectively represented the movement of the actual trawl system. While reproducing the movements of the trawl system using a computer simulation, it is possible to predict the movements of the gear in accordance with the changing parameters related to the gear design and to analyze the gear performance.

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A control system is presented that uses a fuzzy algorithm in controlling the depth of a midwater trawl net, and field experiments carried out on the training ship Kaya of Pukyong National University. The fuzzy controller calculates the length of the warp to be changed, based on the depth error between the desired depth and actual depth of the trawl net and the change ratio in the depth error. The error and the error change are calculated every sampling time. Then the control input (i.e. desirable length of the warp) is determined by inference from the linguistic control rules which an experienced captain or navigator uses in controlling the depth of the trawl net during manual operation. The control input is then transmitted to the trawl winch controller and the length of the warp is changed. The results of these field experiments indicate that this proposed fuzzy controller rapidly follows the desired depth without steady-state error although the desired depth was given in one step, and shows robustness properties against changes in the parameters such as the change of towing speed. The suitability to practical operation of the fuzzy controller designed in this study is fully proved throughout the sea trials.

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In order to evaluate the interaction between handling cost and growth of the bivalve-feeder Halla okudai (Polychaeta: Lysaretidae), the observation of foraging behavior and measurement of energy budget were conducted in laboratory feeding experiments by offering three different bivalves: the short-neck clam Ruditapes philippinarum, the blue mussel Mytilus galloprovinciallis, and the Pacific oyster Crassostrea gigas. The total handling time was 1.6±2.7 times longer in the oyster-group (25.6 ± 9.3 h) than in the clam and mussel-groups, while the total searching and feeding time hardly varied with prey species. No significant difference in the amount of ingestion (4.5±5.3 kcal/polychaete per 15 days) was shown among the three prey groups. However, the growth increment was 1.5±2.1 times higher in the clam-group (1.85 ± 0.65 kcal/polychaete per 15 days) than in the other two prey groups. In contrast, the scope for active rate of respiration and jelly-like material secreted during handling was higher in the oyster-group (0.66 ± 0.14 and 0.24 ± 0.07 kcal/polychaete per 15 days, respectively) and mussel-group (0.62 ± 0.07 and 0.16 ± 0.05 kcal/ polychaete per 15 days) than in the clam-group (0.48 ± 0.09 and 0.06 ± 0.01 kcal/polychaete per 15 days), although there were no significant differences in the standard rate of respiration, excretion, feces and jelly-like material secreted during feeding among the three groups. These results indicate that growth of H. okudai increases with a decreasing handling cost, when feeding on the optimal prey species.

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A suspension-cultured fish cell line was applied to develop a rapid and convenient cytotoxicity assay to evaluate toxicity of a large number of chemicals for fish. Suspension-cultured fish cells, CHSE-sp cells, and monolayer-cultured fish cells, CHSE-214 cells were used. Cytotoxicity of 11 synthesized chemicals chosen from various chemical classes was determined by the neutral red assay. A good correlation was obtained between cytotoxicity values of 11 chemicals to the CHSE-sp cells and to the CHSE-214 cells. Also, good correlations were observed between in vitro toxicity data obtained from the assay using two types of cultured fish cells and in vivo toxicity data obtained from a database on fathead minnow and rainbow trout. From these results, the method using suspension-cultured fish cells used in this study was considered applicable for a screening test method prior to in vivo testing.

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Hemolymph ecdysteroid levels were traced during the molt cycle in female giant freshwater prawn, Macrobrachium rosenbergii. Levels were compared between the reproductive molt cycle, accompanied by ovarian development and oviposition following ecdysis, and the non-reproductive molt cycle, marked by the absence of ovarian development, to examine the involvement of ecdysteroids in ovarian development. Total ecdysteroid levels determined by radioimmunoassay (RIA) showed similar fluctuation patterns between both molt cycles. Levels were low during postmolt stages A-B and gradually decreased to low basal levels at intermolt stages C₀-C₁. Levels then increased during premolt stages D₀-D₂, reached a large peak at the early D₃ stage, and declined sharply prior to ecdysis. In addition, ecdysteroids in the hemolymph were analyzed using high-performance liquid chromatography combined with RIA in order to identify individual ecdysteroid species. During both molt cycles, 20-hydroxyecdysone, ecdysone and high polarity immunoreactive products were detected, and 20-hydroxyecdysone was the most predominant ecdysteroid. Fluctuations in all ecdysteroid species during each type of molt cycle paralleled those of total ecdysteroid levels, and did not show significant differences between the reproductive and non-reproductive molt cycles. These results suggest that ecdysteroids are not involved in ovarian development but in the molting process only.

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Six feeding experiments were conducted to determine energy and protein requirements of different sized yellowtail seriola quinqueradiata (8-280 g) for maximum growth and maintenance of bodyweight under different water temperatures (21.7-27.1°C) in 500 L polycarbonate tanks. For each experiment, fish were fed to satiation or fed at levels between 0 and 80% of the satiation level twice a day. The experimental diets were extruded dry pellets with known digestible energy (DE) and protein (DP) values. The growth rate was highest for the satiation group and decreased proportional to the feeding levels. The DE (kcal/kgBW per day) and DP (g/kgBW per day) requirements for maximum growth, calculated on the basis of the daily feeding rate of the satiation group, were 225 and 21.7, 136 and 14.8, 125 and 11.2, 111 and 10.7, and 92 and 8.2, respectively, for fish with an average initial bodyweight of 8, 63, 160, 237, and 280 g. The requirements for maintenance of bodyweight, based on when the growth rate was zero for the above sizes, were 61 and 5.9, 15 and 1.7, 15 and 1.3, 24 and 2.3, and 12 and 1.0, respectively. The requirements for maintenance of body energy content, estimated on the basis of the relationship between the DE intake and energy retention, were significantly higher than those for maintenance of bodyweight.

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Characteristics in annual reproductive activity of the male golden rabbitfish, Siganus guttatus, were examined by observing testicular histology and measuring plasma levels of steroid hormones, testosterone (T), 11-ketotestosterone (11-KT), and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) by enzyme immunoassay (EIA). The testicular development was divided into four periods: immature (November-March); pre-spawning (April-May); spawning (June-July); and post-spawning (August-October). The immature period was associated with low gonadosomatic index (GSI) values, and the testicular lobules were exclusively occupied by spermatogonia. In the pre-spawning period the testes were characterized by a new generation of spermatogenic cells. Spermatozoa appeared in some testes at the beginning of that period, and became dominant in the testes and the sperm duct in the spawning period. In the post-spawning period, only spermatogonia were observed in the testis, but residual spermatozoa were still observed in the seminal lobules and the sperm duct. Plasma T, 11-KT and DHP levels simultaneously increased during the pre-spawning period, rose markedly at the spawning period, and remained low from the post-spawning to immature periods, suggesting that these changes apparently were related to the annual testicular activity in S. guttatus.

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Head spots of chars were examined for 17 rivers of the Chugoku Region, and relationships between the patterns and water quality were analyzed. To enable quantitative comparison, two spot indices, index I, a ratio of the length to the width for the longest spot, and index II, a ratio of the width of the widest spot to the interorbital width, were calculated. Both spot indices were divided into three ranges, and all char specimens were categorized into eight types, from A to H. Type A with short and wide spots was the most dominant in the Nishiki, Gono and Hii Rivers in the central part of the Chugoku Mountain Chains whereas type H with long and narrow spots was the most dominant in the Gamo River in the eastern part. Electrical conductivity of the water showed a significant positive relationship to index I whereas it showed a significant negative relationship to index II. In contrast, Fe²⁺ concentration of the water showed a significant negative relationship to index I whereas it showed a significant positive relationship to index II.

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To evaluate an interaction between searching cost and growth of the bivalve-feeder Halla okudai (Polychaeta: Lysaretidae), observation of foraging behavior and measurement of energy budget were conducted in feeding experiments for 15 days by supplying two different sizes of prey, Ruditapes philippinarum (small, 24.2 ± 1.5 mm; medium, 34.0 ± 4.1 mm in shell length). The total searching time was longer in the small-prey group (28.9 ± 2.5 h) than in the medium-prey group (16.4 ± 6.4 h), while the total handling and feeding time hardly varied with prey size. The growth rate was higher in the medium-prey group (36.3 ± 5.7%) than in the small-prey group (17.0 ± 9.7%). In contrast, the scope for active rate of respiration and excretion was lower in the medium-prey group (8.5 ± 1.6% and 6.6 ± 0.8%, respectively) than in the small-prey group (20.3 ± 5.5% and 9.4 ± 1.5%), although there was no significant difference in the standard rate of respiration, feces, and jelly-like material secreted during handling and feeding between both prey groups. These results indicate that the differences in searching costs between both prey groups [i.e. the scope for active rate of respiration (11.8%) and excretion (2.8%)] have great effects on the growth (19.3%).

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This study evaluated the contribution of released stocks of ayu, Plecoglossus altivelis, to a target population in the Tohoku area of northern Honshu Island, using allozyme markers. Both natural and hatchery-reared ayu in this area were genetically characterized using 17 gene loci. Samples were taken from one landlocked population from Lake Biwa, from four natural amphidromous populations, and from one hatchery-reared population from Fukushima Prefecture. Two rivers in this region, the Sakari River and the Hienuki River, which had received numerous stocks from all three sources, were examined for stock composition using a maximum likelihood procedure. Significant allele frequency differences were found among the three sources at three of the 17 loci analyzed which are GPI-1*, MDH-1* and MPI* loci. A combination of differences in frequencies of GPI-1*, MPI* and MDH-1* alleles among three released stocks served as natural genetic markers which can identify the source of released stock after stocking. In the Sakari River, which carries an abundant amphidromous ayu population, we observed little influence from either the Fukushima or Lake Biwa stocks. However, in the Hienuki River, which has no native ayu stock, the maximum likelihood evaluated the proportional contribution of released stocks as 36 ± 10% from the Fukushima stock, 42 ± 12% from the Lake Biwa stock, and 22 ± 10% from the amphidromous stock, which corresponds well with the actual proportions of released stock, suggesting that the three released stocks survive and contribute throughout the harvest. The implications of estimates of the stocked contribution are discussed.

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We determined the complete nucleotide sequence of the mitochondrial genome for a Japanese sardine, Sardinops melanostictus (Teleostei: Clupeiformes). The entire genome was purified by gene amplification using long polymerase chain reactions (PCR), and the products were subsequently used as templates for PCR with 30 sets of fish-versatile primers (including three species-specific primers) that amplify contiguous, overlapping segments of the entire genome. Direct sequencing of the PCR products demonstrated that the genome [16 881 base pairs (bp)] contained the same 37 mitochondrial structural genes (two ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes) as found in other vertebrates, with the gene order identical to that in typical vertebrates. A major non-coding region between the tRNA^Pro and tRNA^Phe genes (1200 bp) was considered as the control (D-loop) region, as it has several conservative blocks characteristic to this region.

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A lectin-binding assay was applied to the rainbow trout, Oncorhynchus mykiss, sacculus in order to identify specific sugars in the otolith organ. The binding of biotinylated lectins to the sagitta, otolith membrane and saccular epithelial cells was detected by histochemistry, and their binding to the EDTA-soluble fraction of the otolith matrix and saccular endolymph was detected by dot-blot analysis. N-Acetylglucosamine (GlcNAc) and mannose (Man) were identified in the otolith matrix with both techniques. These data indicate that glycoproteins in the otolith matrix have N-glycosidically linked sugar chains. In the otolith membrane, GlcNAc and Man were identified in the subcupular meshwork, whereas only Man was identified in the gelatinous layer. This is in line with the hypothesis that a part of the subcupular meshwork is incorporated into the otolith as an organic matrix. The saccular epithelial cells reacted with at least one of the lectins which bound to the otolith matrix. The endolymph had the same reactivity with lectins as the otolith matrix. Thus, we suggest that otolith-matrix carbohydrates are produced in the saccular epithelial cells, secreted into the endolymph, and deposited onto the otolith.

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Chionoecetes japonicus is one of the most important commercially fished species in the Sea of Japan. In spite of its economic importance, only limited information on reproduction of this species was hitherto available. Although the reproductive cycle is one of the most important and basic aspects for management, two contrasting opinions have emerged so far: one suggesting a biennial reproductive cycle and the other suggesting a 4-year reproductive cycle. Females of C. japonicus were collected in baited crab pots set on the bottom at depths of 800-1000 m off Jhoetsu, Central Honshu, the Sea of Japan, on nine occasions from April 1993 to April 1994. Carapace width, brood developmental stage and ova size were estimated and analyzed quantitatively. A size frequency histogram showed that most females matured at a modal size of 68.1 mm carapace width, while a small number of crabs matured at modal sizes of either 58.6 or 77.0 mm. Almost all females were multiparous as all but one of the 300 females examined in March had new and/or old ejaculates in their spermathecae. Most multiparous females hatch their larvae and spawn in March and April. Quantitative analysis of ovaries and broods indicated that C. japonicus has a biennial reproductive cycle. The number of eggs in broods varied widely ranging from 0-60 800 eggs, and brood size was correlated with maternal size (P<0.01).

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Reproductive biology of Apogon lineatus in Tokyo Bay, Japan was studied by histological examination of the gonads and measurement of the gonadosomatic indices (GSI). The spawning season occurred during July-October, with both males and females attaining sexual maturity by the end of their first year (60.0 mm TL). It was indicated that A. lineatus is a multiple spawner by the presence of postovulatory follicles and primary-secondary yolk in the same ovaries during the spawning season and the asynchronism of the development of spermatogenesis. Batch fecundity ranged from 7983 to 19 316 eggs for fish of 76.0 mm TL to 106.0 mm TL, respectively, and increased linearly with fish total length. Increases in oocyte diameter were not observed after the tertiary yolk stage, which suggested an adaptation to mouthbrooding, thereby efficiently utilizing the male's restricted buccal cavity to maximize the number of eggs mouthbrooded.

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Demand-feeding behavior of juvenile yellowtails was examined under two different light conditions: in an experimental room and in the outdoors. In the indoor experiment, five groups of 20 juvenile yellowtails (mean bodyweight, 16.0 g) were allocated individually to five 200 L cylindrical tanks set up in the experimental room. A rod-type tactile switch was used for activating the feeding device which delivered 5-10 pellets per actuation. Each tank was illuminated by an overhead fluorescent lamp with a LD 12:12 photoperiod regime. In the outdoor experiment, 69 juvenile yellowtails (mean bodyweight, 79.8 g) were allocated to a 7000 L cylindrical tank. A switch with a string that ends in a rubber, pellet-like knob was used. The feeding device delivered 90-120 pellets by activation of the switch. In both experiments, yellowtail learned demand-feeding within 1-3 days. The number of feeder actuations gradually increased during the experimental period in both experiments, presumably relating both to the rate of learning and to growth of the fish. The yellowtails in the indoor system showed a clear diurnal feeding pattern synchronizing to the given photoperiod, whereas those in the outdoor system showed a clear nocturnal feeding pattern also synchronizing to the outdoor natural photoperiod. Thus, the results showed that yellowtail has an ability for demand-feeding both in light and dark phases. These exact opposite results may have been caused by the difference in intensity of the luminance between the inside of the experimental room and the outdoors.

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A myxosporean parasite was found in the skeletal muscle of porous-head eelpout Allolepis hollandi (Zoarcidae: Perciformes) caught in the Sea of Japan. White pseudocysts, about 3 mm in size, were distributed randomly over the musculature, and gave a negative impact on the appearance of fish products. Fresh spores of the myxosporean were subspherical in shape, 12.0 (11.2-13.5) μm long, 11.0 (10.5-12.0) μm wide, and 7.7 (6.9-8.4) μm thick. Two almost equal polar capsules were 4.8 (3.9-5.4) μm long. A mucous envelope was present around the posterior part of spores. Histological observations revealed that the plasmodia were encased by the host's cartilaginous tissue. Based on spore morphology, tissue-preference and ecological characteristics, we identified the myxosporean as Myxobolus aeglefini Auerbach 1906. This is the first record of a myxosporean infection in porous-head eelpout.

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Solubilization of trimethylamine-N-oxide demethylase (TMAOase) from walleye pollack myofibrillar fraction was examined by various treatments using detergents, acetone, and acid solutions. Without detergent treatments, TMAOase could be extracted from the myofibrillar fraction with 1 M NaCl at pH 4.5, and partially purified by diethylaminoethyl (DEAE)-cellulose and gel filtration chromatography. The specific activity increased 13 000-fold and the yield was 13%, as compared with those in the starting myofibrillar fraction. The enzyme was isolated by means of native PAGE. The partially purified enzyme converted TMAO stoichiometrically to demethylamine (DMA) and formaldehyde (FA), and showed the optimum pH at 7.0. The Km of the enzyme for TMAO was approximately 30 mM. The activation energy was calculated as 38.4 kJ/mol per deg in the temperature range of 0-30°C. The partially purified TMAOase required Fe²⁺ alone for activity. Reducing agents, such as ascorbate, cysteine, and dithiothreitol, were required to maintain the iron in the active form, Fe²⁺. The molecular weight of the enzyme was estimated to be 400 000 by the gel filtration chromatography and to be 25 000 on sodium dodecyl sulfate (SDS)-PAGE. The TMAOase activity was stable in the presence of SDS.

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The permeability of sugar alcohols with different molecular weights (maltitol, sorbitol, and glycerol) into fish meat by soaking was compared. The concentrations of sugar alcohols in the soaked meat increased with an increase in the osmotic pressure of the soaking solution. At the identical osmotic pressure, glycerol gave the highest permeability, followed in descending order by sorbitol and maltitol. In contrast, the moisture content of the soaked meat decreased with an increase in the osmotic pressure of the soaking solution. Maltitol caused the most significant moisture decrease, followed in descending order by sorbitol and glycerol. Comparing the permeability and the dewatering effect in connection with their molecular weights, lowering the molecular weight of sugar alcohols caused an increase in the permeability and a decrease in the dewatering effect. It was concluded that the molecular weight of sugar alcohols had a significant impact on both the permeability and the dewatering effect.

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This study was aimed at evaluating the antioxidant activity of polar carotenoids (zeaxanthin, astaxanthin, and astaxanthin-β-glucoside) in free radical-mediated oxidation of phosphatidylcholine (PC) liposomes. In 1-palmitoyl-2-linoleoyl-PC liposomes, astaxanthin-β-glucoside was the most active, followed by astaxanthin, zeaxanthin, and β-carotene, although there was no significant difference in the antioxidant activity of these carotenoids in chloroform solution. Thus, it is suggested that the different antioxidant activity of four kinds of carotenoids found in the PC liposomes would depend not on their ability to scavenge free radicals, but on other factors such as their location and orientation in PC liposome systems and the degree of their incorporation into PC bilayers. These physicochemical properties would be strongly correlated with the structural acceptability between the PC molecule and carotenoid molecule, especially between two polar groups of the PC bilayer and the carotenoid. Furthermore, two polar groups of the carotenoid can act as trans-bilayer rivets. The tightly packed conformation of PC bilayers caused by this function of the dipolar carotenoid would also be correlated with their antioxidant activity. This was suppported by the result that cholesterol, known as acting like a peg through one half of the bilayer, also increased the oxidative stability of PC liposomes.

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