Fishing efficiency of shrimp trammel net popularly used in Indonesia considerably depends on the area swept by its lower line. The lower line part substituted by a chain was investigated in the present study. As a simple experiment, the chain was towed by a lorry along the hypotenuse of a right triangle drawn on the floor. The tensions of the towed end of chain were measured by a two-component load cell installed in the lorry. The chain curves were photographed by a camera. From measurements of the trajectory of chain movement, the moving angle ø was given as a linear function of the tension angle θ for the parts of chain far from the hypotenuse, ø=αθ+b, and a constant, ø=α (α is the towing angle), for the parts of chain near the hypotenuse. Substituting these equations for the unknown moving angle function in the differential equations, the chain curve and the tension were numerically calculated. When the frictional coefficient was given as μ=μ₉₀ sin²(ø-θ)μ+₀ cos² (ø-θ), the calculated results well agreed with the experimental results. μ₉₀ and μ₀ are the frictional coefficients when the chain was perpendicular and parallel to the moving direction.

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Water temperature effects on foraging responses and thermal tolerances of two congeneric charrs, Dolly Varden Salvelinus malma and white-spotted charr S. leucomaenis, were examined in aquarium settings. Appetites decreased at ≥16°C for Dolly Varden and at ≥24°C for white-spotted charr. The lethal temperatures were ≥20°C for Dolly Varden and ≥26°C for white-spotted charr. The upper limits of preferred temperature ranges and the lethal temperatures for Dolly Varden were significantly lower than those of white-spotted charr. Temperature preference and thermal tolerance probably play a major role in the distribution of these two species.

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To allure squids and prevent them from falling off from jigs, two new kinds of jigs were devised, the impeller-jig and the roller-jig. In the former, the velocity of water flow before the impeller installed in the lure case of the jig was proportional to the diameter of the opening holes on the lure case under a constant winding velocity of 0.6m/s and 1.0m/s. For the latter, the revolution of the roller set at the top of the jig was also proportional to winding velocity, and the roller turned faster at the groove helix angle of 30° than that of 20° and 45°. The results showed that the new jigs were useful and effective to allure the target squid and hold them firmly with the visual stimulation of the rotating parts.

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The spawning cycle of Calliurichthys japonicus and Repomucenus huguenini was investigated from March 1992 to February 1993 in a wild population in Tosa Bay, Kochi Prefecture. Calliurichthys japonicus is an annual spawner, having high reproductive activity in summer. Thespecies appears to spawn mainly between 90-120m water depth and extends its spawning ground to shallower waters of the Bay during the peak reproductive period in July. Maturity of females occurs at 12.5cm in SL and 17.7cm SL in male. Repomucenus huguenini is a biannual spawner, spawning withinits normal distribution range of 15-30m water depth during spring (May) and autumn (October to December with a peak in November). This species matures and reproduces from an early age with maturity estimated to occur at 4.5cm SL in both sexes. The temperature in 45-120m when the Gl of C. japonicus samples was at its peak in July, ranged from 17.4-21.6°C and salinity from 34.6-35.0. During the peak reproductive season of R. huguenini in November, the temperature ranged from 22.4-22.8°C and salinity from 34.4-34.6 in 15 to 30m water depth.

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Growth of the barnacle Balanus albicostatus was investigated for the population in an aquarium at 20.0-30.5°C from March to the following February. For feeding, the diatom Chaetoceros calcitrans and Artemia nauplii were given. Shell size in the basal diameter of the rostro-carinal axis and mortality were examined monthly. Mean molting cycle was estimated from the daily frequencyof the molting for 45 days. Shell strain during molting growth was characterized by a strain gauge method.
The mean±S. D. of shell size showed 4.5±1.7mm in March and increased to 10.7±2.4mm in the following February. The daily growth rate ranged from 0 to 29 μm/mm•day duringthe experimental period. It tended to be high until September, and decreased thereafter. The mortality in the period was 18.3%. The molting cycle of the population was 5.0 days at 28°C. The shell showed compressive stress at the intermolt, and its strain decreased during molting.

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Feeding experiments were conducted at 20°C with diets containing 0, 9, 18, 36, and 44% of meat and bone meal (MBM) to examine the potential of MBM as a substitute for fish meal in the diet of Japanese flounder. Juvenile fish, 2 to 5g in initial body weight, were fed on each diet to satiation twice a day, 6 days per week, for 4 and 7 weeks (trials 1 and 2).
In both trials, the final body weight of fish, the weight gain, feed efficiency (FE), and protein efficiency ratio (PER) of fish fed MBM diets tended to decrease as the proportion of MBM in the diet increased. The FE of fish fed the diet containing 44% of MBM as well as PER of those fed 36 and 44% MBM diets were significantly lower than those of the control diet (0% MBM diet). The weight gain of fish fed 36 and 44% MBM diets was also lower than that of the control in trial 2. No significant differences in these parameters were observed among dietary groups containing 0, 9, and 18% of MBM except the final body weight in trial 2. There was little difference in the proximate composition of the whole fish body and the hematological and hematochemical parameters between the fish fed 18% MBM diet and the control group.

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The artificial induction of gynogenesis of amago salmon was cytologically confirmed, and the nucleus behavior and the cytological rates were compared at the early stage of development among the following three groups: normal, hybrid between the female amago salmon and the male rainbow trout, and gynogenetic developed group. The first cleavage of hybrid and gynogenetic development was delayed compared with normal development, while that in the gynogens was later than that of the hybrid. The cytological stage at 68h•°C in cumulative temperature, which is considered the optimum timing for suppressing the first cleavage of gynogen, was prometaphase. In gynogenetically activated eggs, the lump-shaped body appeared in the metaphase plate. Then, it remained near the cleavage plane in the following cell division, and was considered to be degenerated nuclear materials derived from irradiated sperm, i.e. dense chromatin body (DCB), which could not transform to chromosomes. Therefore, paternally derived genetic materials should disappear during the anaphase and the telophase in the first cleavage of gynogenetic development. Some chromosomes were gradually eliminated in the hybrid development after the first cleavage. This phenomenon was considered a major cause of inviability and the subsequent expression of abnormality during the development of the hybrid.

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In order to investigate the early life history of yellowtail Seriola quinqueradiata, age composition in the schools of yellowtail associated with drifting seaweeds was analyzed by otolith daily increments. Increments of sagittal otolith was verified as daily rings through the examination of reared specimens from hatching to day 60. Mean age and birthdate of ten schools (n=291; TL, 18.1-113.8mm) collected in the East China Sea during the period from 19-21 May 1992, ranged from day 39 to 67, and from 12 March to 10 April 1992, respectively. Seven of ten groups showed approximately uniform age composition, because 70% of fish in a school were included within 4 days range of each mode. It is speculated that yellowtail spawned at the same time and same place formed a school and migrated until early juvenile stage associated with drifting seaweeds. Growth rate of reared specimen was higher at an early juvenile stage (ca. day 60) than that of wild specimen.

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Effects of water temperature and light intensity on aggressive behavior were examined in the juvenile yellowtails Seriola quinqueradiata (16-32mm TL). When fish were acclimated to water temperatures at 15, 20, 25, and 30°C, frequency of aggressive behavior increased with the increase in water temperature. Under the light conditions of total dark, 10³, and 10⁴ lx, the highest aggression was observed at 10³ lx, whereas no aggression at dark. No aggressive behavior was observed at 10^-3 lx whereas weak aggression occurred at 10^-2 lx, suggesting the threshold of light intensity between 10^-3 and 10^-2 lx.

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The frictional force acting on the vertical synchronous belts used in hauling upthe net or rope and the main factors affecting the frictional forces were determined through experimental and theoretical analysis. The frictional force was calculated by using Euler's equation. It was shown that the average frictional coefficient between belt and net was 0.78 and that between belt and rope was 0.70. When the net or rope was wound out by belts along a spiral line, the frictional force wasfound to decrease with an increase in the spiral angle. At a constant velocity, the speed of moving up the net or rope had no effect upon the frictional force. On the other hand, a non-uniform velocity of moving reduced the actual frictional force.

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A survey of the occurrence and characteristics of Bdellovibrio around the coastal areas and aquacultural ponds of Taiwan was conducted. The recovery rate of bdellovibrios was highest in estuaries during summer. Prey-dependent Bdellovibrio sp. strain TS-S and strain TS-V were isolated from the estuary of the Tam-shui river, from which prey-independent strains were further selected by streptomycin treatment or by using enriched medium. Their mutation rates were 1.2×10^-6 (streptomycin method) and 3.8×10^-6 (enriched medium) for the strain TS-S and 1.1×10^-5 (streptomycin method) and 2.1×10^-6 (enriched medium) for the strain TS-V. Four prey-independent mutantswere isolated from strain TS-S by the enriched medium method and designated as Bdellovibrio sp. strains TS-S_PI1 through TS-S_PI4. Similarly, two prey-independent mutants, designated as TS-V_PI1 and TS-V_PI2, were obtained from strain TS-V. The physiological characteristics of these six prey-independent mutants differed from those of the three species (Bdellovibrio bacteriovorus, B. stolpii, and B. starrii), in comparisons by biochemical tests and electron microscopic examination. The average cell sizes of TS-S_PIs and TS-V_PIs, were 0.18×0.85 and 0.21×0.78 μm, respectively. These sizes are smaller than the cell sizes of the three species shown in Bergey's Manual.

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We investigated the yearly fluctuation of survival indices before and after shirasu (post larval Japanese sardine) stage, from 1979 to 1992, comparing the relative abundance of shirasu with the annual egg production and with the year-class strength of Japanese sardine off thePacific coast of Japan. The relative abundance of shirasu was estimated, using the catch data from the shirasu boat seine in Enshu-nada and Atsumi-gaikai area, off Shizuoka and Aichi Prefecture, and the location of Kuroshio Current axis. Between egg and shirasu stage from 1979 to 1985, and between shirasu stage and recruitment from 1979 to 1987, the survival indices were high on average though widely fluctuated. However, the former indices after 1986 and the latter indices after 1988 were consistently low. Except after 1988 when both stages' survival indices were low, it was observed that the survival index before the shirasu stage is higher than that after the shirasu stage is low and vice versa.

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Antisera raised against piscine pituitary hormones and heterologous antisera against mammalian pituitary hormones were used to analyze the types and distribution patterns of cells in the adenohypophysis of pejerrey Odontesthes bonariensis. Prolactin (PRL) cells were immunostained in the external border of the rostral pars distalis using anti-chum salmon PRL and anti-carpPRL. Immunoreactive (ir-)corticotrophic (ACTH) cells were identified in the same area using anti-human ACTH and crossreactivity was observed in the pars intermedia (PI), corresponding to the presumptive melanocytestimulating hormone cells. Ir-growth hormone (GH) cells were identified in closecontact with the neurohypophysis in the proximal pars distalis (PPD) using anti-chum salmon GH and anti-carp GH antisera. Ir-gonadotrophic (GtH) cells were distributed in the PPD and in the external area of the PI. Two forms of GtH were identified using antisera raised against chum salmon GtH Iβ and GtH IIβand anti-croaker maturational GtH. Ir-thyroid-stimulating hormone (TSH) cells were revealed with an anti-human TSHβ antiserum in the PPD. Ir-somatolactin (SL) cells were detected in the PI using two antisera raised against chum salmon and red drum SL.

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Exposure to a high concentration of salinity caused a great increase in blood osmotic pressure and death of the guppy. To elucidate the strain differences of osmoregulation in seawater, blood osmotic pressure was measured after transfer from freshwater to 15 ppt dilute sea water in 13 different strains of the guppy. Blood osmotic pressure increased and peaked at 24h after the transfer and then decreased to a level comparable to that in freshwater. Although the blood osmotic pressure infreshwater did not differ among strains, the values 24h after transfer to 15 ppt dilute sea water demonstrated that the blood osmotic pressure was largely different among strains. A significant inverse correlation was observed between the sea water tolerance (mean survival time in 35 ppt sea water) and the blood osmotic pressure at 15 ppt dilute sea water. This suggests that the strain differences in sea water tolerance depend on differences in osmoregulatory function.

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A full-length cDNA for medaka cytoskeletal β-actin was isolated from a liver cDNA library. The entire coding region was 1125 bp long and was well conserved with respect to other vertebrate β-actins. Two primers for a reverse transcriptase-polymerase chain reaction (RT-PCR) assaywere constructed from the nonvariable region among medaka, carp, chicken, and mouse β-actin. RT-PCR amplified a 510 bp DNA band from medaka cDNA synthesized from the blood, brain, heart, intestine, kidney, liver, muscle, ovary, spleen, and testis poly(A)+mRNA.
The constructed primers were used successfully to detect a specific 510 bp fragment of β-actin from zebrafish, rainbow trout, and Japanese flounder.

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Locomotor activity in the rainbow trout Oncorhynchus mykiss was recorded at 10°C using an infrared photocell system. Under light-dark cycles of 12:12 h (LD 12:12), all fishexhibited diel rhythms with higher activity during the light phase. When fish were exposed to constant conditions such as constant darkness or constant dim light, fish exhibited free-running rhythms of locomotor activity with τ in a circadian range. When fish were subjected to phase shift of LD 12:12 cyclesby doubling the light phase or the dark phase, resynchronization to the new LD cycles was attained notimmediately but a few cycles of transient phase were observed. These results indicate that diel rhythms of locomotor activity in the rainbow trout are strongly influenced by a circadian clock entrained to LD cycles.

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Morphological development of veliger larvae and juveniles of the hatchery-rearedsurf clam was examined. Larvae settled by 0.26mm shell length (SL) with the gills and the siphons remaining scarcely differentiated. Although the inner demibranch began to develop just after settlement, the outer one did not do so until about 1.3-1.5mm SL. After settlement, the primary exhalant siphon arose at first by 0.3mm SL. The inhalant siphon appeared between 0.4 and 0.6mm SL, and the exhalant one between 2.2 and 2.5mm SL. Although distinct in early juveniles, the byssal gland degenerated by 5.0mm SL. Digestive organs developed rapidly, once a shell length of 3-5mm SL was reached.
The juvenile stage consists of four substages that differ in their mode of life.Newly settled juveniles depend on interstitial seawater because of the lack of the inhalant siphon. Development of the inhalant siphon enables the juveniles to draw in seawater lying above the bottomsurface. Elongation of the siphons enables the juveniles to burrow deeper into the seabed. Juveniles almost completely attain the adult features and mode of life by 15mm SL.
The organs do not develop harmoniously in the surf clam. Some morphological changes tend to trigger transitions in the mode of life before there has been sufficient development of organs and functions required for the subsequent life style. Such inharmonious organogenesis probably results in deficiencies in adaptation at the transitions.

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The present study examined the pharmacokinetics of oxytetracycline in rainbow trout Oncorhynchus mykiss. The pharmacokinetics of oxytetracycline were determined after intravenous administration (50mg/kg body weight). Serum concentrations of oxytetracycline were determined using high performance liquid chromatography with direct injection. Serum levels could be fitted to a two-compartment model. The calculated half-lives for the distribution phase (T_1/2α) and the elimination phase (T_1/2α) were 0.5 and 52h, respectively. The apparent volume of distribution (V_dβ) and total body clearance (Cl_B) were calculated to be 1.46 l/kg and 19.6ml/kg/h, respectively. The area under the serum concentration-time curve was 2554 μg•h/ml. Serum protein binding in vivo of oxytetracycline was 51.1±7.4%. As an application of this pharmacokinetic study, the oral bioavailability and the dosage regimens of oxytetracycline in rainbow trout were briefly evaluated.

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Twenty eight strains of the marine bacteria which killed the marine dinoflagellate, Gymnodinium mikimotoi, were isolated during the occurrence of the huge red tide by G. mikimotoi in Tanabe Bay in 1990. Among them 22 strains belonged to the genus Vibrio, three to Flavobacterium, two to Acinetobacter and one to Pseudomonas-Alteromonas. All of them grew in filter-sterilized natural seawater without any nutrient supplements, suggesting that the killing bacteria are ubiquitous inseawater and are related to the occurrence of G. mikimotoi opportunistically. Most of the killing bacteria did not affect the growth of three marine diatoms, Skeletonema costatum, Ditylum brightwellii and Thalassiosira sp., and some of them did not affect the growth of the marine dinoflagellate, Alexandrium catenella. These findings strongly suggest that the killing bacteria influence the change of microalgal community in a marine environment.

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To obtain more basic and detail information on the ecology of marine sulfate-reducing bacterial population, we studied the use of in situ hybridization with 16S rRNA probes for their cellular level detection from natural environments. SRB probe was found to have wide specificity to sulfate-reducing bacterial groups. SRB probe hybridizes also with most species of δ-group of purple bacteria. However, most of them were found not to dominate in the coastal and estuarine environments. Although two species, Clostridium novyi and Myxococcus xanthus, have same sequences with SRB probe, they can be eliminated through observation under fluorescent microscope with having endospore and autofluorescence, respectively. Desulfobacter probe was found to be specific to genus Desulfobacter. We modified dual staining method of Hicks et al. (1992) with SRB probe, and compared the numbers of sulfatereducing bacteria enumerated by the present method with conventional MPN method (medium containing sodium lactate) using microcosm of zooplankton decomposition. The numbers (4.5×10⁶-2.7×10⁷/ml) detected using in situ hybridization with SRB probe were about two orders of magnitude higher than MPN method (1.1×10³-1.5×10⁵/ml).

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Microcosm experiments were performed to know the dynamics of sulfate-reducing bacterial popu-lation and volatile fatty acids (VFA) during decomposition of polypepton and detrital material in the surface sediments from Tokyo Bay and Sagami Bay. Population of sulfate-reducing bacteria were detected and counted through fluorescent in situ hybridization with SRB probe which has wide specificity to sulfate-reducing bacterial group, and with Desulfobacter probe which has narrow specificity only to genus Desulfobacter.
In the microcosms of the Tokyo Bay sample, vibrioid (0.7-1.0×2-2.5μm) and rod (1.5-1.8×2.5-3.5μm) shaped SRB were detected on day 3, SRB population changed to cocci (0.7-1.0μm) and rod (1.5-1.8×2.5-3.5μm) shaped cells on day 8. These rod shaped SRB were hybridized also with Desulfobacter probe. The increase of Desulfobacter followed by the increase of utilized acetate. In the Sagami Bay sample, only rod shaped SRB (0.7-1.2×2.5-6μm) was detected on day 35. On day 100, cocci (2-3μm) and short rod (0.7-1.2×1.7-2.2μm) shaped SRB were detected. No cell was hybridized with Desulfobacter probe.
It was considered that there was the succession of sulfate-reducing bacterial population during decomposition of detrital material in the surface sediments. The probe method would be helpful to understand population changes of sulfate-reducing bacteria in the natural bacterial communities.

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To determine which protein bands appeared in SDS-polyacrylamide gel electrophoresis (PAGE) patterns of sarcoplasmic proteins (Sp-P) are responsible for the gel strengthening effect of Sp-P on the myofibrillar protein (Mf-P) gel, the relations between the jelly strength of Mf-P gel added with each of three different sarcoplasmic fractions and the relative amounts of protein bandsin the SDS-PAGE pattern of each sarcoplasmic fraction were examined.
The heat-coagulability of each sarcoplasmic fraction was highly correlated with the jelly strength of each gel. Relative amounts of 94, 64, and 40 kDa components in the SDS-PAGE pattern of each sarcoplasmic fraction also showed a good correlation with the jelly strength of gel. These results suggest that the high jelly strength of Sp-P added Mf-P gel is attributable to a large amount of heat-coagulable proteins, especially the 94, 64, and 40 kDa components of Sp-P.

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A 148 kDa protein which possibly participates in collagen metabolism was isolated from the adductor muscle of the abalone Haliotis discus, by a series of chromatographic procedures involving DEAE-Toyopearl 650 M ion exchange, TSK G3000SWG gel filtration, and Butyl-Toyopearl 650 M hydrophobic columns. Reactivity with collagen was measured as the ability to solubilize the film containing pepsinsolubilized collagen from the same animal. Active fractions of various purification steps solubilized the film at 37°C within 60min. The purified protein consisted of a single polypeptide chain and the molecular weight was estimated by SDS-PAGE to be 148, 000. This protein did not containmethionine and half-cystine, and its N-terminal amino acid sequence differed from any sequencesof known collagenases.

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Changes in chemical constituents and enzyme activities were assayed in red sea bream Pagrus major through embryonic development from eggs immediately after fertilization to larvae just before feeding. No significant changes were found in protein, total nitrogen, and phospholipid contents with proceeding development stages. Although free amino acid (FAA) and triacylglycerol (TAG) contents continuously fell, DNA and RNA contents reversely increased through the development. Ammonia, free glucose, and glycogen contents were maintained at markedly low levels. Aspartate aminotransferase, alanine aminotransferase, creatine kinase, and alkaline phosphatase activities showed a similar change through the development, decreasing or remaining constant at low levels up to the stage of Kupffer's vesicle appearance and increasing steeply thereafter. These results suggest that endogenous TAG and FAA are preferentially expended as energy reserves under the embryonic development of red sea bream. The rapid increases of the enzyme activities after the stage of Kupffer's vesicle appearance might correspond to their organ differentiation and function.

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Changes in fatty acids and prostaglandin E₂ (PGE₂) concentrations in the acetic acid treated red al-gae Gracilaria (G.) asiatica and G. rhodocaudata samples were investigated. Fatty acid concentrations increased in lipid, particularly phospholipid and free fatty acid classes of ogonori treated with 4% acetic acid at 37°C and heated at 100°C. In the acetic acid treated ogonori, free arachidonic acid (AA) significantly increased, but PGE₂ decreased. These results suggested that the enhancement of AA would be due to the depression of PGE₂ formation from AA with the acetic acid treatment. Fatty acids concentrations varied according to species of ogonori. AA concentration of G. asiatica, acausal species of human poisonings which occurred in Japan, showed higher levels in both the raw andacetic acid treated than that of G. rhodocaudata.

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The glass transition in protamines, mainly salmine (from salmon), in the presence of maltose or dextran has been studied using differential scanning calorimetry. The water activities (Aw) of salmine and its mixtures with maltose or dextran were adjusted from Aw 0.11 to 0.93 and state diagrams showing relationships between glass transition temperature (Tg), Aw and water content were established. The adsorption isotherms and Tg-water content relations were obtained by applying the Guggenheim-Anderson-Deboer equation and the Gordon-Taylor equation, respectively. Salmine used in this study was in a glassy state (Tg=about 140°C at 0% water content) and its Tg value decreased with increasing water content. The Tg values of salmine were reduced in the presence of maltose, while increased by mixing with dextran. The Tg of the maximally freeze-concentrated phase of salmine (Tg') was found out to be -5.8°C.

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To investigate structure-function relationship for akazara scallop troponin C (TnC), we constructed a bacterial expression system using a cDNA clone which we isolated previously (Ojima, T., Tanaka, H., and Nishita, K. Arch. Biochem. Biophys., 311, 272-276 (1994)). The cDNA for akazara scallop TnC was subcloned into the expression plasmid pET-16b and expressed in Escherichia coli BL21 (DE3) by the induction with isopropyl-β-D (-) -thiogalactopyranoside. Amount of the expressed TnC increased up to approximately 10% of total protein during 4h incubation. The TnC was thenextracted from the bacterial pellet and purified by subsequent column chromatographies on DEAE-Toyopearl 650M and Phenyl-Sepharose CL-4 B. Although the expressed TnC possessed an extra moiety of 6 amino acids at N-terminus, which was derived from adaptor DNA of λgt11 and 5'-untranslated region of the cDNA clone, it showed basically the same properties as those of native TnC with respect to the Ca²⁺-induced difference UV-absorption spectrum and Ca²⁺-regulatory ability when reconstitutedwith native tropo-nin T and troponin I.

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The aqueous extract from the mature roe (gonad index>10%) of the lamprey Lampetra japonica was found to be lethal to mice when injected intravenously, while that from the immature roe(gonad index about 4%) was nontoxic. Dinogunellin, the well-known fish roe toxin, was notdetected in the lamprey roe. The lamprey toxin was a thermolabile, basic protein and purified to an electrophoretically pure state from the mature roe by chromatography on CM-cellulose and hydroxyapatite, FPLC on Mono S, and HPLC on TSKgel Ether-5PW. The purified toxin was highly lethal to miceupon intravenous injection; its LD₅₀ was estimated to be 13 μg/kg. Analyses by SDS-PAGE either with or without a reducing agent showed that the toxin has a molecular mass of 36 kDa composed of two kinds of subunits with molecular masses of 28 and 10 kDa. The amino acid composition of the toxin was characterized by the abundance of Ser, Glx, and Gly and the scarcity of half-Cys.

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