The investigation was conducted to develop the prospects for application of cryopreservation technique to nauplius larvae of economically important crustacean, such as Japanese prawns or crabs. Since Balanus amphitrite possesses naupliar stages similar to those of prawn, the nauplius-II larvae of barnacle were applied in this experiment. The larvae were equilibrated for 20 minutes in a cryoprotectant solution containing 1.5M dimethyl sulfoxide (DMSO) in 29‰ filtered seawater. They were then frozen to-196°C by a two-step freezing procedure. The optimum conditions for the cryopreservation of the nauplius-II larvae were: from ice seeding temperature of-12°C to the first step cooling temperature of-30°C was by a cooling rate of 0.5°C/min, then rapidly frozen to-196°C. Frozen nauplii were thawed at 50°C, and incubated in 29‰ filtered seawater after washing cryoprotectant. The diatom Chaetoceros calcitrans at a concentration of 40×10⁴ cells/ml was provided as food in the rearing of nauplii. After 28 days preservation in liquid nitrogen (-196°C), 78% of the larvae recovered and 19.8% of the larvae metamorphosed to cyprid.

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Sequence analyses of small subunit ribosomal RNA gene (16S rDNA) were performed on 16 strains of gliding marine bacteria isolated at the time of bloom of Skeletonema costatum in Ariake Sea. The strains were comprised of 8 strains which have capabilities to kill and lyse Skeletonema costatum, 5 strains with algicidal or algal-lytic activity and 3 strains without activity. The strains were divided into 4 clusters including 1, 2, 3, and 10 strains. Cluster A, C and D were close to Cytophaga latercula, Flexibacter maritimus and Cytophaga marinoflava, respectively. Cluster B was close to Flavobacterium aquatile. Neither algicidal nor algal-lytic activity was restricted to any of the clusters. The strains of cluster B contained flexirubin and did not require addition of Na⁺ in growth peptone media.

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Chattonella marina and Heterosigma akashiwo are highly toxic red tide phytoplankton which causes severe damage to fish farming. Recent studies demonstrated that these flagellates release superoxide anion (O₂^-) under normal growth conditions. In the present study, we examined the effects of the fish mucus prepared from skin and gill of yellowtail, red sea bream, and japanese flounder on the production of ₂^- by the flagellates. All mucus tested stimulated C. marina and H. akashiwo to generate increased amounts of O₂^- in a concentration dependent manner. Thus, the common substances present in fish mucus of three species may be responsible for the stimulation of the flagellates. In the presence of 100μg/ml of each mucus, 4-6-fold increased level of O₂^- was detected in both flagellates. After the heat treatment (at 121°C for 15min), no significant decrease of stimulatory effects of the mucus was observed, suggesting that heat-stable components of the mucus may produce the stimulatory effect on the flagellates. Our results suggest the possibility that the fish mucus enhances the oxygen radical-mediated toxic effects of C. marina and H. akashiwo.

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The hearing sensitivities of bastard halibut Paralichthys olivaceus decreased sharply at the frequency band between 200Hz and 400Hz. The hearing response of bastard halibut was measured by sweeping sound which the center frequency moves from non-hearing frequency (300-1600Hz) to hearing frequency (100-200Hz), and discussed the upper limit frequency of hearing.
The classical conditioning was reinforced by the tone burst sound in a continuous time of 1s, a frequency of 100Hz and a sound pressure level of 131dB. The sweeping sound was projected in five steps at intervals of 5dB. The sweeping speed of eight types was adjusted with a combination of a start and end of sweeping frequency. The conditioning response was measured as suppression in the respiratory activity by a piezoelectric sensor fitted on the gill cover.
The response to sweeping sound was 82% at the frequency band between 200Hz and 340Hz, which agreed with the boundary of the hearing area. The upper limit frequency of hearing of bastard halibut existed under 500Hz. The hearing response to sweeping sound did not agree with the audiogram of bastard halibut because the sound pressure level fluctuated in the small water tank. The sweeping speed is inversely proportional to the individual difference of the response time.

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We compared tolerance of three aquaculture fishes (yellowtail, red sea bream and Japanese flounder) to Chattonella marina, and studied physiological responses to exposure to the alga. Resting oxygen uptake (Mo₂) was also determined for the three species. All yellowtails were killed by exposure to C. marina, whereas mortality was 33% and 0% for bream and flounder, respectively. Arterial oxygen pressure (Pao₂) declined rapidly in yellowtail and bream soon after the onset of the exposure. The Pao₂ gradually fell and remained lower than the control values in two flounders, whereas it was close to the pre-exposure levels in the other four flounders at the end of the exposure. Ventilatory response during C. marina exposure was consistently intense in yellowtail, variable among individuals in bream while nearly absent in flounder. Resting Mo₂ was highest in yellowtail, and lowest in flounder. We consider that the observed difference in tolerance to C. marina resulted from the different oxygen requirements of the species and their physiological responses in hypoxia (oxygen regulator/conformer). It appears that venous oxygen extraction as a means to maintain oxygen uptake during hypoxia does not function efficiently in fish with a higher oxygen requirement like yellowtail.

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In order to estimate the genetic differentiation of natural populations of the snake-head fish in Thailand, 12 enzymes were examined in 13 populations collected from the north, the central, and the northeast region. Seventeen enzyme loci were detected, and polymorphism at 6 loci was revealed. The characteristic frequencies of alleles were regionally observed at IDHP-2^* and MPI^* locus. The genetic distances among populations within region were smaller than among populations of 3 regions. The average genetic distance among populations between the north and the northeast region and between the central and the northeast region were larger than that between the north and the central region clearly. The populations could be divided into 2 groups of the north-central and the northeast population on the basis of the dendrogram drawn by genetic distances. The genetic groups coincided with the geographical groups divided by 2 large river systems in Thailand.

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Partial nucleotide sequences in the exonic region of a small subunit ribosomal RNA gene (ssr DNA) were analysed for nine commercially utilized species of Porphyra and eight laver cultivars in order to examine whether these sequences can be used to identify and discriminate between these species and cultivars. Pairwise comparisons of a total of 555 nucleotides revealed that there were from one to ninety-one nucleotide differences (substitutions and insertions/deletions) among the nine species. The one nucleotide difference found between Porphyrayezoensis and P. tenera was authentic, confirmed by repetitive sequence analyses of ssr DNA exonic regions using thalli of these species collected from five different locations. The nucleotide sequences of eight laver cultivars were identical to each other and also identical to field-collected P. yezoensis but not P. tenera. These results suggest that the nine Porphyra species can be distinguished from each other by comparison of the partial nucleotide sequences of the ssr DNA exonic region, while discrimination of laver cultivars is not possible by the comparison of only these sequences.

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Six strains of Tetraselmis, comprising T. tetrathele, T. chui, T. alacris, T. gracilis, T. suecica Spain, and T. suecica Brasil, were cultured in seawater (salinity, 27‰) supplemented with 80mg of ammonium sulfate, 16mg of calcium perphosphate and 4mg of Clewat-32per liter, with or without 100mM glucose, under 14h illumination (3, 000 lx) per day. All the strains except T. alacrrisgrew better in the medium containing glucose. In particular, the growth of T. tetrathele and T. chui was accelerated by the addition of glucose, the rates being 1.42 and 1.40 doublings per day, respectively, at the logarithmic phase. When cultured in the dark, their growth rates ranged between 0.94 and 0.90 doublings per day in the presence of glucose. An appropriate condition for the mixotrophic culture of T. tetrathele was considered to be as follows: 100mM glucose, 0.6mM ammonium sulfate, 10h/day illumination, and 30°C. Under this condition the protein, carbohydrate, lipid, and fatty acid contents of this alga were 31, 11, 13, and 4.5%/cell dry-weight, respectively. Furthermore, the growth of rotifers was enhanced by feeding cells of glucose-grown T. tetrathele.

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To characterize genetically both natural populations and cultured strains of the guppy (Poecilia reticulata), enzyme polymorphism was examined by starch gel electrophoresis. There are three kind of natural populations: wild populations, natural populations which have originated from the introduction of wild guppy and reversion to the wild state of the domesticated guppy (introduced populations). While the cultured strains have originated from the guppy which have been maintained in pet shops and established as strains.
The screening of genetic variation at 30 loci revealed that the introduced populations were significantly higher than the cultured strains in the amount of genetic variability. The quantification of genetic differentiation revealed that the cultured strains were more diversified than the introduced populations.
The genetic variability of the introduced populations was the similar level in comparison with the wild populations of Trinidadian guppy. The degree of genetic differentiation of the wild Trinidadian populations was significantly more diversified than the introduced populations of the present work.
The decrease of genetic variability and increase of genetic differentiation would be caused by founder effect at the division of the population.

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Heritabilities, and genetic and phenotypic correlations were estimated for total length, body weight, body depth, and head length of masu salmon Oncorhynchus masou. During 4 month period after first starting to feed in juveniles, 20 full-sib and 10 half-sib families were reared separately and each family was given environmental conditions as equal as possible. Data were collected at 4 months and 8 months of age by individual tagging with fin cut 1994. The same experiments using colored fluorescent elastomer tags as family markers were repeated in 1995. Overall survivals in the study varied among each family. The heritabilities of growth related traits estimated for sire components (in 1994 and 1995) at 4 months age were found to be quite high: ranging between 0.35 and 0.47 in total length. Estimated heritabilities at 8 months age in each year ranged from 0.18 and 0.42 in total length. Similar values were obtained in the other three related areas of body weight, body depth and head length. Genetic and phenotypic correlations among the growth traits were all positively high. Genetic variations in body weight were found to be remarkably high, about three times as high as that in the total length of the masu salmon. Phenotypic correlations between 4 and 8 month ages were 0.45-0.63. Selective breeding in growth related traits would be effective in masu salmon culture.

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Seasonality of reproduction in sporophytes of Laminaria japonica was investigated on the basis of the nitrogen content of thalli cultivated in the field in southern Hokkaido, Japan. Fertile plants occurred throughout most of the year. Sorus formation was greatest in the basal parts of thalli in fall (September). The secondary peak of sorus formation occurred in the apical parts of thalli in winter (February and March). The seasonality of sorus formation was linked closely to the nitrogen content in the thalli. The nitrogen content per volume of thalli declined from January to April and then increased to a maximum in September. In January and February, the nitrogen content was higher in the apical part(ca. 2mg•cm^-3) than in the basal part of thalli. After June, however, it was higher in the basal part. Sorus area increased in association with the nitrogen content per volume of thalli regardless of the thallus part or season. In addition, although sorus formation took more than 100 days after April, it needed only about 50 days in February. These results suggest that the low nitrogen content in summer delayed sorus formation and decreased the area of the sori. The onset and position of sorus formation depended on the growth and the nitrogen content per volume of each thallus.

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Effects of 17α-methyltestosterone (MT) immersion treatments at selected doses and durations on sex reversal of the mud loach Misgurnus mizolepis were examined. Fry were immersed in water containing MT 0, 25, 50, 100 or 200μg per l for one, two or three weeks. Survival decreased as dose and treatment duration increased; in most instances it was significantly lower than that of the controls (p < 0.05). Growth rate at all the treatment combinations did not differ, when compared to that of control (p > 0.05). The treatment with 200μg MT l^-1 for two or three weeks produced all male populations, and the group treated with 100μg l^-1 for three weeks also became nearly an all male population (99.3%). The morphological and histological attributes of testes of the hormone-treated fish were similar to those of normal males. The hormone treatment altered the secondary sexual character of the fish,
namely the size and morphology of the pectoral fins.

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Mass mortalities of cultured sea urchin Strongylocentrotus intermedius occurred at Date, Shiriuchi and Shikabe Fisheries Breeding Centers in Hokkaido, from the end of May to the beginning of June (seawater temperature ranged 11_??_13°C) in 1995. A short rod psychrophilic bacterium, which has a single polar flagellum was isolated from coelomic fluid and body surface of diseased animals at the three Centers. The isolates were Gram-negative, oxidase positive, and acid production from glucose anaerobically without gas production. The G + C contents of DNAs from the representative strains Da-2 (Date), Sr-3 (Shiriuchi) and Sk-1 (Shikabe) were 42.7, 43.5 and 44.3 mol%, respectively. On the basis of these characteristics, it was presumed that the isolates belong to the genus Vibrio. Experimental infections with sea urchin were performed by immersion method using 10⁷, 10⁶and 10⁵ CFU/mlof the representative strains Da-2, Sr-3 and Sk-1 at 11°C. Groups, which were injured mechanically on the body surface showed higher mortalities than that of the uninjured group. Especially, the group injured on the connective layers around the teeth showed the highest mortality. These results suggested that the isolated bacterium is the causative agent of the disease of sea urchin Strongylocentrotus intermedius occurring at low water temperatures.

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Age and growth of the cardinal fish Apogon lineatus in Tokyo Bay were estimated from right sagittal otoliths of 556 fish. Examination of the outer margins of the otoliths showed that the translucent zone was formed once a year, and can be used as an annulus, with its formation completed at the beginning of the spawning season. Growth of fish, expressed by the von Bertalanffy growth equation, was L_t=86.6[l-exp{-1.12(t+0.01)}] for males, and L_t=118.5[l-exp {-0.37(t+1.03)}] for females, where L_t is total length in mm and t is age in years. Growth of males and females differed. Females were larger than males at each estimated age. Most fish examined were ages 1 and 2, although the oldest fish were 3 years old for both sexes.

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The eth hemolysin gene locus of Edwardsiella tarda encodes the hemolysin EthA protein and its accessory protein EthB. We constructed mutants by site directed marker insertion mutagenesis in ethA or ethB by a method employing the integration of a mobilizable suicide plasmid containing a portion of the ethA or ethB coding sequence into the chromosome. Both the ethA- and ethB-matants lacked extracellular, cell-associated, and intracellular hemolytic activities. These results indicate that the EthB protein is essential for the hemolytic activity of EthA. The EthA protein may be activated in the periplasmic region of the cell by the EthB protein which is located on the outer membrane of the cell. The loss of hemolytic activity of the ethA- and ethB-deficient mutants suggests that E. tarda has only one hemolysin gene locus in the genome.

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Acoustic target strengths of captive juvenile fishes of three species were measured at four frequencies using fish cage method.
Measurements were conducted on a barge constructed at a fishing port for the experiments. Measured fish were sea smelt Hypomesus pretiosus japonicus (Brevoort), goldeye rockfish Sebastes inermis (Cuvier), and purple puffer Takifugu porphyreus (Temminck et Schlegel). They were caught with a miniature purse seine and maintained in fish breeding tanks before the experiments. Their body lengths ranged from 30 to 65mm. Each species was classified into three size classes. Specimens were maintained in a cubical cage of 0.5m each side made of lace cloth during measurement. The echo signals from encaged fish at 25, 50, 100, and 200kHz were recorded with a data recorder. Echo signals were later digitized and processed by squared integration to calculate the target strengths.
The results show that, (1) The backscattering strength of encaged juveniles indicates a linear density dependence for most sample sets, (2) The normalized target strengths TScm of juveniles ranged from -66 to -55 dB, which is maximum 10 dB higher than that of adults, (3) Target strength changed by the frequency, that is, the TScm for L/λ_??_2 was roughly 10 dB higher than the TScm for L/λ_??_2.

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Soybean trypsin inhibitors (SBTI) inhibited trypsin and basic proteinase in the hapatopancreas, pyloric ceaca and intestine of cultured-marine and freshwater fishes. The highest trypsin inhibition by the SBTI was obtained in tiger puffer Takifugu rubripes, followed by rainbow trout Oncorhynchus mykiss, striped jack Pseudocaranx delicatissimus, red sea bream Pagrus major, Japanese striped knifejaw Oplegnathus fasciatus, grouper Epinephelus septemfasciatus, bluefin tuna Thunnus thynnus, purplish amberjack Seriola dumerili, yellowtail Seriola quinqueradiata. The highest basic proteinase inhibition by the SBTI was obtained in red sea bream, followed by rainbow trout, grouper, purplish amberjack, tiger puffer, Japanese striped knifejaw, bluefin tuna, striped jack, yellowtail. These results suggest that the dietary soybean meal utility, being somewhat higher in a rinbow trout, red sea bream, and tiger puffer than yellowtail, do not actually correlate to their sensitivity of trypsin-like and basic proteinase for SBTI.

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Many studies on fishery management assume that the objective of the fishery management is to maximize their economic rent and the fisherman operates to maximize their respective net profits. In order to confirm these assumptions, this paper tries to evaluate the net profit and the marginal net profit of the scoop net fishery in Lake Biwa, which is a main fishery for ayu Plecoglossus altivelis. Here, the net profit is defined as the landing in value terms minus the operating cost. This paper calculates the net profit per boat and the marginal net profit per boat using two cost functions. One includes only the variable cost, and the other is the opportunity cost including the estimated family wages.
The net profit and the marginal net profit have the negative value specially at the end of the fishing season of the scoop net fishery, using a cost function including the estimated family wages. This does not support the above assumptions. This suggests that the fisherman's behavior depends not only on the net profit but also on other factors in the scoop net fishery. This is due to the less adjustability of labor power in the fishery. Under this economical condition of the fishery, it is difficult to maximize the economic rent as an objective of the fishery management.

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Developmental changes in the visual acuity of red sea bream were investigated by histological examination of the retina for 61 individuals from juvenile to adult stage (BL 23-609mm). The highest cone density could be located in the dorso-temporal area of the retina. The visual acuity depends both on the eye lens diameter and the cone density on the retina. The previous study reported that the minimum separable angle is obviously greater than the resolving power of the lens. The lens diameter increased from 1.3mm to 11.8mm proportionally with the growth of body length. The cone density was found to fit in an exponentially declining curve, where it steeply decreases in the range of 23-100mm BL and becomes a gentle slope to be around 200-400 cells/0.01mm² for samples over 100mm BL. According to the results, the visual acuity was increased with growth; from 0.05 for 23mm BL to 0.28 for 504mm BL specimen.
The improvement of acuity can be attributed increase of the lens focal length rather than of the cone density. The visual acuity (V.A.) was shown to fit to the allometric function of the body length (BL) as follows,
V.A.=0.00711BL^0.588, (r²=0.964).

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The effects of a three week intramuscular implantation of gonadotropin-releasing hormone analog (GnRHa) on gonadal maturation and changes in serum gonadal steroid hormone levels were examined in anadromous, maturing sockeye salmon Oncorhynchus nerka during the pre-spawning season. Spermiation occurred in both GnRHa-implanted and control males, but serum 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) levels in the implanted males were higher than in controls. No differences were found in serum testosterone (T) levels between GnRHa-implanted and control males. Serum 11-ketotestosterone (11KT) levels in control males increased for three weeks, but those in GnRHa-implanted males did not change. All GnRHa-implanted females ovulated completely for three weeks, while 25% of control females ovulated partially and the rest of females ovulated 17-24 days after sampling. The number of ovulated eggs of GnRHa-implanted females was greater than that of control females, and percentages of eyed eggs were not different between GnRHa-implanted and control females. Serum levels of DHP increased dramatically in GnRHa-implanted females, but those of T and estradiol-17β (E₂) was similar to control females. These results confirm that GnRHa accelerates gonadal maturation of both sexes through elevation of serum DHP levels in maturing sockeye salmon.

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In order to investigate the composition of extractive nitrogenous constituents in the olive flounder Paralichthys olivaceus cultured by recombinant bovine somatotropin (BST) supplemented-feed and the BST unsupplemented-feed (control) at the south coast of Korea, the muscle was analyzed for extractive nitrogen, free amino acids, oligopeptides, nucleotides and related compounds, quaternary ammonium bases, and guanidino compounds using specimens collected from April 1996 to March 1997. Free amino acids such as taurine, alanine, serine, threonine, lysine, cystathionine, and glutamic acid were rich in all the samples. Among them taurine was the most abundant, occupying from 9 to 38%. As for nucleotides and related compounds, inosine 5'-monophosphate was the principal constituent in all the samples. β-Alaninebetaine, homarine were detected in all the samples but glycinebetaine was found only in some samples. The large amount of creatine was detected in all samples from 305 to 545mg/100g muscle. There was no conspicuous difference in the extractive nitrogenous constituents between the two group of samples when compared with the amount of free amino acids, nucleotides and related compounds, bataines, trimethylamine oxide (TMAO) and trimethylamine (TMA), and creatine and creatinine, except the results of oligopeptides and sensory evaluation (p<0.10).

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Frozen walleye pollack surimi blocks were thawed and heated at 30°C for up to 120min to induce thermal denaturation. The heat-treatment decreased an endogenous TGase and myofibrillar Ca-AT-Pase activities to 14% and 37% of the initial values, respectively. The surimi gel formability and setting response were almost completely lost. Although the addition of MTGase to the heat-denatured surimi completely recovered myosin cross-linking reaction, the gel strength and setting response were only partially recovered. In the presence of EGTA, an inhibitor of the endogenous TGase, the gel formability and setting response of the native surimi paste were remarkably reduced. However, these properties were completely recovered by the addition of MTGase together with EGTA. These results apparently demonstrate that the maximum gel strength of a final gel reflects the development of an elastic gel network which formed by a two-step heating procedure depending on a combination of thermal gelling ability of native myosin molecules and TGase reaction in the surimi paste.

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Full-length cDNA for a 70.6 kDa heat-inducible heat shock protein (HSP70), a member of the HSP70 family, was isolated from a cDNA library of cultured cells in early passage originated in Japanese flounder embryos (JFE cells). It has a single ORF of 1920 bp that encodes a protein of 70.6 kDa. Japanese flounder HSP70 contains an EEVD peptide motif at C-terminal end which is a common feature of the cytosolic HSP70 family. Japanese flounder HSP70 is 86.0%, 83.8%, and 83.6% identical in primary structure to chinook salmon HSP70, bovine HSP70, and human HSP70, respectively. Japanese flounder HSP70 is also 83.6% identical to Japanese flounder HSC71. These results suggest that the amino acid sequence of the cytosolic HSP70 family, HSP70 and HSC71, has been highly conserved among vertebrates. Northern blot analysis showed that HSP70 mRNA was induced with heat shock treatment in the JFE cells in early passage.

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Atka mackerel meats cured with 0.5-2.0M NaCl containing 20mM Tris-acetate (pH 7.0) were incubated at 15°C, 30°C and 50°C and in 60% relative humidity for 0-16 hours, and their drying characteristics and textural change during drying at different temperatures were simultaneously investigated. Slow moisture vaporization occurred at the initial drying period and the critical moisture content significantly decreased with an increase in the NaCl content of cured meats. In addition, the reduction of the drying rate in the later drying period was suppressed when 0.5mol/kg of NaCl were contained in the cured meats. Furthermore, at 15°C and 30°C drying, the increase in the shear force of dried products with the decrease in the moisture content was effectively suppressed by the curing with NaCl. Such changes in the drying characteristics and texture would contribute to depression of the excess hardening and obtaining a long shelf-life of dried products.

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Water soluble proteins (WSP) from the flesh of blue marlin Makaira mazara were modified through the Maillard reaction with glucose (G) or glucose-6-phosphate (G6P) by heating at 60°C for up to 12h. The development of brown color, loss of free amino groups, formation of fructosamine, and amount of bound phosphorus were determined. The Maillard reaction of WSP + G6P was found to progress faster than WSP+G. The incorporation of phosphorus into WSP took place mainly during the early stage of the Maillard reaction. From the results of SDS-PAGE analysis, it was revealed that the polymerization of proteins was also accelerated in WSP+G6P. Increasing phosphorus content of WSP brought about the improvements of solubility at pH above 7 and thermal stability of WSP. WSP modified with G6P at 60°C for 3 to 6h retained protein solubility more than 80% even after heating at 100°C for 10min.

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Post-mortem changes in content of ATP and its related compounds, free amino acids, and polyamines were investigated in the tail muscle of Japanese spiny lobster Panulirus japonicus, rock lobster Jasus lalandii from South Africa, and American lobster Homarus americanus from Canada during storage at 0, 5, and 10°C.
Initial ATP level in the muscle of Japanese spiny lobster was higher than other species and the decreasing rate was the slowest. During storage, AMP and IMP accumulated to a similar extent in the muscle of Japanese spiny lobster, while only IMP accumulated in the muscle of American lobster. In the muscle of rock lobster, neither AMP nor IMP accumulated and inosine increased rapidly. Taurine, asparagine, glutamine, proline, glycine, and arginine were dominant in every lobster, and no polyamine was detected in the early stage of storage. With degradation of arginine, ornithine increased in the muscle of rock lobster, while both ornithine and putrescine increased to about the half of the initial arginine level in the muscle of American lobster. Arginine remained at a higher level, and the other two compounds did not increase in the muscle of Japanese spiny lobster.

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