We designed five 16S rRNA-targeted oligonucleotide probes (Sp probes) specific for Flavobacterium sp. 5 N-3, which inhibits the growth of red tide phytoplankton Gymnodinium mikimotoi (Dinophyceae). These probes were evaluated by whole-cell hybridization against 5 N-3 cells incubated under laboratory conditions. The fluorescence signal from the cell detected with Sp probe mix5, a mixture of the five probes, was 8.4-fold higher than that obtained with only one Sp probe (Sp01RF). The signal obtained by this method was strong enough to recognize 5 N-3 cells and count them under the epifluorescence microscope, while the signal was often undetectable when a single probe was used. Fluorescence intensities of cells at stationary phases and of ‘starved’ cells in sterile seawater using Sp probe mix5 were low but still sufficient for enumeration. These Sp probes did not hybridize with 11 strains from the Cytophaga/Flavobacteria/Bacteroides phylum and did react with strain 5 N-3 following whole-cell hybridization. These results show that 5 N-3 cells cultivated under our laboratory conditions can be detected by whole-cell hybridization with the five designed probes. These data also suggest that this technique may be useful for detection of an algicidal bacterium 5 N-3 in the natural environment.

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The correlation between hematology and plasma chemistry was examined in natural silver crucian carp Carassius langsdorfii. Three microsatellite DNA markers identified five clonal lines. Blood was obtained from the caudal vessels and analyzed using automatic analysis systems. The correlation between several parameters including erythrocyte count (RBC), mean corpuscular hemoglobin concentration (MCHC), alkaline phosphatase (ALP), plasma glucose (Glu) and total protein (TP) was assessed by liner regression. Hemoglobin (Hgb) and hematocrit (Hct) as a function of the erythrocyte count, hematocrit as a function of the hemoglobin, mean corpuscular hemoglobin (MCH) as a function of the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) as a function of the mean corpuscular hemoglobin, and total protein as a function of triglyceride (TG) were positively correlated for the five clonal lines. Different regression slopes (b-value) of RBC to Hgb and Hct were found among clonal lines, while no such differences were found among slopes of the remaining regression equation. This phenomenon suggests that the significantly different slopes among parameters, which indicated low genetic influence at the each parameter, clearly detected the genetic influences as the compound effect of these parameters.

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In order to characterize the spontaneous green-type pigmentation mutant of Porphyra yezoensis, growth and contents of photosynthetic pigments were compared with those of the wild type in gametophytic blades. The growth of the green mutant was slower than that of the wild type. The content of phycoerythrin was markedly lower in the green mutant than in the wild type. For genetic analysis, the green mutant was crossed with the wild type. In the cross, the heterozygous conchocelis (the wild-type color) produced many sectored F₁ gametophytic blades, which were composed of both parental colors. The segregation ratio by counting the wild-type sectors and the green-type sectors of the F₁ blades was approximately 1:1. This suggests that the green mutant is governed by a single recessive gene. All the monospore germlings from the sectored F₁ gametophytic blades that originated from the heterozygous conchocelis developed into single-colored blades with either the green color or the wild-type color. The advantages of using heterozygous conchocelis (hybrid conchocelis) and monospores from F₁ gametophytic blades that originated from the heterozygous conchocelis in commercial farming of Porphyra yezoensis are detailed and discussed.

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To reveal the ontogeny of pancreatic exocrine function in the early larval stage of eel, cDNAs encoding major pancreatic enzymes, trypsinogen, amylase and lipase were identified from the Japanese eel Anguilla japonica and their expression pattern in larvae was analyzed. The cloned eel trypsinogen precursor consisted of 224 amino acids and showed 82.2% identity to trypsinogen-2 of winter flounder Pleuronectes americanus. The eel amylase precursor consisted of 512 amino acids and showed 77% identity to winter flounder amylase. Eel pancreatic lipase was composed of 470 amino acids and had 58.3% of identity to human pancreatic lipase. In the eel larvae, mRNA expression of trypsinogen and amylase was first detected at 6 days post-hatching (d.p.h.), and the expression level increased between 7 and 8 d.p.h. In contrast, mRNA expression of lipase was first detected at 8 d.p.h. Eel larvae start to feed actively at 8 d.p.h. Thus, it was indicated that eel pancreas starts to synthesize digestive enzymes at 6 d.p.h. and acquires full function by the onset of exogenous feeding at 8 d.p.h.

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To quantify the contribution by cocultured animals to waste assimilation in an intensive shrimp farm in Thailand, the food web structures of the macrobenthos in a reservoir pond, a shrimp culture pond and water treatment ponds were examined using the stable C and N isotope ratio technique. Seawater for aquaculture was drawn from a creek, and stored in a reservoir pond, used for farming the banana prawn Fenneropenaeus merguiensis in culture ponds, and then recycled through treatment ponds where the green mussel Perna viridis was cultured to remove organic wastes discharged from the farming. The clam worm Nereididae sp. and the mud creeper Cerithideopsilla cingulata in the culture pond had &¹³C values of -21.0‰ and -18.4‰, respectively, suggesting that shrimp feed (mean &¹³C=-20.7‰) was the main food source for these species. The &¹³C analysis also suggested that sediments (-23.7‰) in the reservoir pond and particulate organic matter (POM) (-24.0‰) and/or sediments (-25.0‰) in the treatment pond supplied carbon for most macrobenthic animals. However, green mussels in the treatment pond had a mean &¹³C value of -20.5‰, suggesting that shrimp feed was the main food source for this species.

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The mechanism of deciding a posture of the trolling depressor in the towing was investigated. The hydrodynamic characteristics of the trolling depressor, which has been widely used by Japanese coastal fishermen, were investigated using models with a 394.5 cm² plane form area in different shapes and cambers. These models were rigged for an attack angle from -10° to 90°. The lift, drag and moment were measured by a three-component load cell in the circulating water tank. The difference in the shape of the trolling depressor did not affect the drag coefficient. Superior efficiency of the trolling depressor is considered to be due to the effect of camber. The position of the pressure center very near the front edge makes the trolling depressor move downward. Disagreement of points between the towing point and the pressure center accelerated the swinging movement of the trolling depressor.

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For the development of a stepwise cryopreservation technique for larvae of the Pacific oyster Crassostrea gigas, various conditions were examined. Larvae at 9, 12, 15, 18 and 21 h after insemination were cooled at a rate of −1°C/min (seeding at -8°C for 15 min) and then plunged into liquid nitrogen at -35 or -40°C using 1.5 M dimethyl sulfoxide (DMSO) and 250 mM trehalose as cryoprotectants. Among these larvae, 15 h after insemination (the trochophore stage before formation of the shell gland) showed the highest motility and the best external appearance after thawing. Trochophore larvae were cryopreserved in preservation media containing different dilutions (1/4, 1/6, 1/8, 1/10 and 1/30) of seawater. Larvae preserved in the 1/4 seawater medium showed the highest appearance of shelled larvae 4 days after thawing. Trochophore larvae reared in seawater at 21, 25 or 29°C were cryopreserved for 8 months and then reared at 26°C after thawing. Larvae reared at 25°C showed the highest survival rate and normal larval ratio at day 6 after thawing, although larvae reared at 21°C showed the highest rates until day 4. One larva developed at 25°C succeeded to settle.

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Lethal limits of high temperature were studied to clarify the effects on the survival of the endangered Japanese crayfish species Cambaroides japonicus and the alien species Pacifastacus leniusculus. After the acclimation period for 2 weeks at 16°C, the temperature was raised at a rate of 1°C per week. As a result, the ultimate upper lethal temperatures of C. japonicus and P. leniusculus were 27.0 and 31.1°C, respectively, and the lethal temperature for P. leniusculus was significantly higher than for C. japonicus. The natural distributions of these two species are discussed in terms of the temperature tolerance.

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Attempts have been made to assess the end-point temperature (EPT) of heated fish and shellfish meats by using the coagulation method together with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and enzyme activity determination. Unfrozen and frozen fish and shellfish meats were heat-treated at different selected temperatures with 0, 15 or 30 min holding times. Proteins were extracted with NaCl solution. The coagulation method was able to determine EPT of heated fish and shellfish meats between 60 and 67°C. SDS-PAGE patterns of the filtrates from heated meats were closely related to the results of the coagulation method and enzyme activity determination. Two proteins responsible for producing coagulum of fish meat extracts seem to be lactate dehydrogenase (LDH) and glyceraldehyde phosphate dehydrogenase (GAPDH). End-point temperatures determined by these methods were not significantly different between unfrozen and frozen samples. On the contrary, a highly thermostable protein with a molecular mass of 35 kDa was detected in heated shellfish meats up to 108°C. In scallop adductor muscle, this highly thermostable protein was found to be the tropomyosin subunit from its amino acid composition and their partial sequences. Tropomyosin could be used as an EPT indicator up to 108°C for heated shellfish meats.

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The filamentous fungi Rhizopus, like many fungal species, possesses physiologically active substances. Rhizopus extract (RU) is reported to be effective for various aspects of growth and reproduction in many vertebrates. The effects of RU administration on body growth and plasma levels of steroid hormones were investigated in lacustrine sockeye salmon Oncorhynchus nerka. One-year-old fish were fed daily with RU (20 mg/kg feed) from July 1999 to October 2000 for 15 months. Fish were sampled every month and plasma levels of testosterone (T), 11-ketotestosterone (11-KT), estradiol-17β (E₂) and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) were measured by time-resolved fluoroimmunoassay. Body growth of RU-fed fish of both sexes increased significantly in 1⁺ and 2⁺ October, and 2⁺ January-March and July. All RU-fed males and one female matured in 2⁺ October. RU-fed 1⁺ precociously mature males showed increased plasma levels of T, 11-KT and DHP in 1⁺ October. In 2⁺ males, RU significantly elevated plasma levels of T from May to June, 11-KT from June to July, and DHP in October. In sockeye salmon, administration of RU accelerated body growth of both sexes and sexual maturation in males, suggesting physiologically active substances present in RU enhance somatic growth and sexual maturation by sex-specific mechanisms.

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The present study examined the changes in texture and protein components during cold storage of different squid varieties. Raw oval squid, Japanese common squid and arrow squid were sliced fresh and the muscles were stored at 4°C for 0, 4, 8, 12, 18, 24, 48 and 120 h. The rheological measurements, protein components and amounts of collagen were examined. The adhesiveness of each squid increased significantly in the early stage of cold storage. In all varieties, penetration decreased at 4 h, which is considered to be rigor mortis, then increased. The amounts of total collagen, 20°C water-soluble collagen and 70°C water-soluble collagen did not change significantly in each variety during cold storage. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern showed that the 580 kDa component gradually disappeared up to 48 h. The correlations between the amounts of 580 kDa component and adhesiveness or firmness were high. Models of fit based on chemical kinetics accurately expressed the behavior of adhesiveness, firmness and penetration showing that 63.2% of adhesiveness changes occurred in 13-19 h and that 63.2% of firmness changes occurred in 18-24 h.

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The genetic structure and variability of four wild populations of kuruma prawn in Japan were examined by estimating relatedness among individuals. The relatedness was estimated by five microsatellites (MS) DNA markers. Examination of relatedness showed that individuals were related significantly in Kumamoto and Kagoshima. In Kagoshima, some individuals showed full-sib level of relatedness. The analysis of mitochondrial (mt) DNA polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) was also performed, showing that the closely related individuals in Kagoshima tended to share a common haplotype. It is, therefore, supposed that there are many kins in Kumamoto and Kagoshima. However, the heterozygosities and allelic and genotypic frequencies in MS-DNA analysis were not significantly different among the localities. Moreover, the haplotype distributions of mtDNA in Kumamoto and Kagoshima were significantly different from other localities. Thus, it is suggested that no spatial differentiations occurred due to the geographic or historical effects between the localities and that there is the possibility of a mixture of hatchery populations in Kumamoto and Kagoshima.

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The effects of smallmouth bass on the behavior, growth and survival of the prey fish (goldfish, ayu and Japanese dace) were examined experimentally in artificial ponds. In most cases, goldfish aggregated both when smallmouth bass were present and absent. Ayu and Japanese dace aggregated more frequently in the presence of smallmouth bass. All prey fish fed predominantly on benthic algae or aquatic insects and their dietary composition was not affected by smallmouth bass. In goldfish and Japanese dace, the total number of feeding acts exhibited by each individual was significantly less in the presence of smallmouth bass than in the absence and, in goldfish, the growth rate was less when smallmouth bass were present. In contrast, the presence of smallmouth bass did not affect either the total number of feeding acts or the growth rate of ayu. However, ayu were preyed on by smallmouth bass most frequently among the three prey species. Smallmouth bass tended to prey on smaller individuals, but ayu were not smaller than goldfish or Japanese dace.

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A non-linear finite element method is employed to determine the equilibrium configuration and tension distribution of a net set in a uniform current. The method models each mesh bar in the net as an infinitely flexible rope element connected at each end to other elements by a frictionless hinge. The Newton-Raphson method is used to solve the basic non-linear simultaneous equations with respect to tension in mesh bar and knot displacements. Global convergence to the correct solution from an initially estimated state is ensured using a loading iteration in addition to the common configuration iterative procedure in order to accurately model the shape-dependent characteristic of hydrodynamic forces. The method is shown to predict the shape and tension distribution of fishing nets with reasonable accuracy in the case examined in the present study.

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The requirement for taurine in juvenile Japanese flounder Paralichthys olivaceus was determined by feeding diets containing various levels of taurine and cystine. Test diets supplemented with 0.5, 1.0 and 1.5% of taurine or with 0.5, 1.0 and 1.5% of L-cystine were prepared. The basal diet contained 55% protein from white fish meal. These diets were fed to juvenile Japanese flounder with an initial mean bodyweight of 0.9 g (total length (TL) 48 mm) for 5 weeks. Approximately 1.4% taurine content in the diet was required for optimum growth of juvenile flounder. A positive linear relationship was noted between the content of taurine accumulated in the muscle, liver and brain and the level of taurine in the diet. However, there was no increased taurine content in tissues of fish fed the cystine-supplemented diet. In contrast, the fish fed control and cystine-supplemented diets showed higher contents of cystathionine in the tissues. The concentration of cystathionine in tissues rapidly decreased with an increase of taurine in the diet. It was also observed that for each of the dietary groups, a trace amount of taurine was excreted. These results suggest that the taurine content in the diet affects the sulfur amino acid metabolism of juvenile Japanese flounder, and indicate that juvenile flounder are unable to biosynthesize taurine from cystine.

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During spermatogenesis, many proteins are synthesized in the testis prior to the completion of sperm maturation. Many components are involved in the testicular protein synthesis and one of the components that is implicated in the polypeptide chain elongation is elongation factor 1α. In the present study, the molecular cloning of elongation factor 1α (EF-1α) was conducted from a testis cDNA library of the Nile tilapia. The cDNA for tilapia EF-1α (tEF-1α) contains a complete open reading frame encoding 462 amino acids. The predicted amino acid sequence of EF-1α shows an approximate 90% similarity to those identified in other teleost fish, such as medaka, sea bream and zebrafish. Northern blotting revealed that the gene is expressed in all of the examined tissues and in ovulated eggs. The results of in situ hybridization indicate that the gene is expressed specifically in Leydig cells in testis, suggesting the involvement of EF-1α as an actin-binding protein in the cluster formation of Leydig cells. In the ovary, the gene is expressed in the perinucleolus stage of oocytes, suggesting that EF-1α is also implicated in oocyte growth.

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Effect of nonylphenol (NP) products from different companies on in vitro vitellogenin (VTG) synthesis was compared using tilapia (Oreochromis mossambicus) hepatocytes cultures. Addition of NP at a concentration of 10^-3 M to the medium caused death of hepatocytes (NP-A and NP-B) and delay of monolayer formation (NP-C). No cell death was observed at a concentration of 10^-4 M (NP-A, -B and -C) but cell adhesion was slower than control. These results suggest that high concentration of NP is toxic against tilapia hepatocytes. When effects of estradiol-17β (E₂, 10^-7 M) and NP (10^-4 M) on in vitro VTG synthesis were examined, addition of E₂ and NP-A and NP-B to the media resulted in elevation of VTG. NP-C did not induce VTG in the medium. Co-treatment of NP-B and tamoxifen, a non-steroidal anti-estrogen, reduced VTG synthesis. These results suggest that NP has estrogenic potential in primary cultures of tilapia hepatocytes and acts through binding to the estrogen receptor, and that there is a difference in the induction level of VTG among different NP products.

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A growth model for the endangered cyprinid fish Tribolodon nakamurai was derived following otolith analyses of 16 wild and 53 reared specimens. The asteriscus was the most appropriate to measure size among three otolith elements, and its height OH mm was used as size index of otolith. Standard length L cm was best back-calculated using the Gompertz model, L=70.0·exp[-exp{-0.553 (OH-2.73)}]. Translucent zones on the lapilli, analyzed from 5-year-old-reared fish, were regarded as winter slow-growing zones. The ages of 10 wild specimens of 37.0-48.1 cm standard length were calculated as 7-10 years by counting the translucent zones on the lapilli. Age t was best back-calculated using the allometry model, t=1.33·OH^1.37. The growth trajectory of T. nakamurai followed a slender S curve, three typical growth models, von Bertalanffy, Logistic and Gompertz, and Richards' model, which is a general formula of the above three, being fitted using the maximum likelihood method. The Gompertz model, L_t=60.2·exp[-exp{-0.258(t-4.68)}], was found by Akaike's information criterion (AIC) to be the statistically most acceptable growth model.

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We investigated stress responses of ayu Plecoglossus altivelis under the hypothesis that overcrowding during transport can be a stressor and the stress predisposes fish to disease by decline in immunocompetence. The extent of stress response was determined under different fish densities. As inferred from the serum concentration of cortisol 1 day after the treatment, the density at more than 23 kg/m³, which was within the range of normal transporting procedures, acutely evoked intense stress responses. To examine a possible way to reduce the stress response of fish under crowding conditions, we compared two fish groups established in water tanks with circulating (whirling) and turbulent currents at the same density. The cortisol levels 1 day after the treatment were 2.5-fold higher in turbulent current than in whirling current. Fish in whirling current formed a school, while those in turbulent current aggregated with each other. Schooling of fish induced by water flow may reduce their stress responses during transport. A transport system with a directional water flow should keep fish in good condition without reducing the number of fish contained.

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Onset and development, and diel rhythm of aggressive behavior were observed in the early life stages of Japanese flounder Paralichthys olivaceus. The frequency of nip was counted as an index of aggressive behavior. No aggressive behavior was observed during the larval stage until day 39 after hatching (when fish were 13.2 mm in standard length (SL)) in the experimental tanks. The onset of aggressive behavior was observed on day 39, coinciding with completion of metamorphosis from the larval to the juvenile stage, and it developed until day 46. During the metamorphosing stage, typical shivering behavior (Ohm (Ω)-posture) was observed, and it showed a peak on day 19 (SL 7.6 mm) and decreased thereafter. Ω-Posture disappeared in the juvenile stage. Aggressive behavior and Ω-posture were only observed in the daytime. Settlement was first observed from day 19 in less than 30% of larvae and all individuals settled from day 39. As Ω-posture was observed regardless of the presence or absence of food and the peak of frequency in Ω-posture was observed in a specific stage of metamorphosis, this behavior is not related to feeding behavior, and it is possibly a precursor to aggressive behavior in this species.

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Gustatory response of marbled rockfish Sebastiscus marmoratus was recorded from the facial nerve supplying the anterior palate. In amino acids, the lowest threshold was for L-proline at 10^-6-10^-5 M; in ATP-related substances, the value for inosine was approximately 10^-5 M and in organic acids, the value for L-lactic acid was approximately 10^-6 M. An almost complete self-adaptation was observed for stimulants after 5 s application. While a weak solution of L-lactic acid as adapting stimulus depressed the response to it completely, it did not appreciably affect the response to 10-times or more concentrated L-lactic acid solution. In contrast to a weak solution, a moderate solution caused a severe depression of the response to stronger solutions, resulting in a shift of the dose-response curve towards a high concentration range. Time for recovery from adaptation differed depending on chemicals and stimulus duration: for 10^-4 M L-lactic acid, it took about 2.3 and 4.5 s for 1/2 recovery of response magnitude under unadapted condition from the adaptation after 1 s- and 5 s-stimulation, respectively. L-Lactic acid and L-alanine strongly depressed the response to each other, yielding a rightward shift of the dose-response curve. A similar profound cross-adaptation was also observed between inosine and 5'-inosinic acid. The possible role of gustation is discussed.

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Seasonal reproductive and lunar-related spawning cycles of the female honeycomb grouper Epinephelus merra inhabiting Okinawan waters were examined by histological observation of ovaries. Gonadosomatic index (GSI) increased beginning in May and peaked in June. Histological observations revealed that many oocytes laden with yolk were present in the ovaries from May to August. From September to October, ovaries were occupied by immature oocytes. These results suggest that the reproductive season of E. merra lasts for 4 months from May through August in Okinawan waters. When the fish were collected according to the lunar cycle, GSI increased with the approach of the full moon. Oocytes at various development stages were observed from the first quarter to the full moon. Fresh ovulatory follicles were present in the ovaries around the last quarter moon. These results suggest that E. merra has a lunar spawning cycle and spawns between the full moon and the last quarter moon. Ovarian features of the fish collected around the last quarter moon were different among individuals; some fish had many oocytes at the tertiary yolk stage in the ovaries, while the ovaries of the others were occupied by the oocytes at the peri-nucleolus and the oil droplet stages. This observation suggests that a minor release of eggs occurs in this species before or after a major spawning lunar day. Oocytes at the migratory nucleus and the maturation stages were not observed in any ovarian samples. This may mean that maturation of oocytes is related to the spawning behavior of this species and makes rapid progress in the process of aggregation at the spawning sites.

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The effect of dietary digestible protein/digestible energy (DP/DE) ratios and feeding level on growth, feed efficiency, nutrient and energy usage by Atlantic salmon (Salmo salar; initial body weight, 7.0 g/fish) at 15°C was investigated in a 16-week feeding trial. Three diets, differing in their DP and DE contents, namely 37/18 (regular diet, RD), 37/21 (high fat diet, HF) and 44/ 22 (high nutrient-dense diet, HND) g/MJ of dry feed were formulated. DP/DE ratios were 20, 18 and 20 g/MJ for the RD, HF and HND diets, respectively. Salmon were hand-fed three times a day at either 100% or 85% of the feed requirement estimated by a bioenergetics model. At each feeding level, DE intake (kJ/fish) was similar for all three diets. Diet composition did not affect growth rate. However, increasing the digestible energy density from 18 to 22 MJ/kg of dry feed resulted in a significant increase (P<0.05) in feed efficiency. Restricting feed intake significantly decreased live body weight gains for all diets. However, feed efficiency was not affected by feeding level. Diet composition and feeding level did not affect carcass composition and nutrient and energy usage, with the exception of a higher (P<0.05) carcass lipid of fish fed the HF100 diet compared with the fish fed the RD and HND diets and a higher (P<0.05) lipid gain (g/fish) of fish fed the HF100 diet compared with fish fed all the diets at the restricted feeding level. Restricting feeding resulted in significantly lower (P<0.05) energy gain (kJ/fish) compared with fish fed at 100%. Increasing the DE and nutrient density of the diet had no effect on growth but improved feed efficiency and lowered solid wastes (g of solid wastes per kg of fish produced) while dissolved wastes were not affected by dietary ormulation.

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Functional sex reversal of all-female nigorobuna Carassius carassius grandoculis to phenotypic males was examined by immersion exposure of fry to 17-methyltestosterone (17-MT) and controlled water temperature during early development. Fry were reared in water containing different concentrations of 17-MT at 24 and 30°C for 80 days starting 20 days after hatching. Although the fish exposed to 0.1 and 1.0 μg/L 17-MT at 24°C were all male, treatment with 10.0 μg/L 17-MT resulted in 43% females. Twenty-two percent males appeared in the control treatment at 30°C but the control at 24°C was entirely female. The proportion of males in treatments exposed to 0.01-1.0 μg/L 17-MT at 30°C was slightly lower than in the respective treatments at 24°C. These results indicate that the phenotypic expression of sex in nigorobuna is thermolabile and that sex determination is under the control of genetic factors and temperature. Also, control of temperature during early development has been shown to be important for the production of all-female offspring for use as breeding stock for pond culture of fish suitable for preparation as ‘funazushi’.

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A study was conducted to examine the variation in fish farm sediment quality with varying organic carbon input in the form of feed. In order to elucidate the seasonal variation in farm sediment quality, relation between seasonal organic carbon input, temperature and sediment quality was studied. The sediment ignition loss, redox potential and acid volatile sulfide sulfur (AVS-S) content showed a correlation with annual organic carbon input. The seasonal AVS-S content showed a significant correlation with the seasonal organic carbon input as well as temperature. Multiple regression analysis revealed that both of these factors explicated 80% of the seasonal variation in AVS-S content. Although redox conditions of the sediment could also be correlated with both the organic carbon input and temperature, the latter seemed to have a greater control. Thus, the farm sediment quality could be directly related to the annual organic carbon input to the fish farm under similar hydrographic conditions. Acid volatile sulfide sulfur content exhibited a significant correlation with both the annual as well as the seasonal organic carbon input. Therefore, AVS-S content proved to be a fairly simple and better indicator for monitoring the fish farm environment for effective farm management and sustainable aquaculture. Importance of size of pellets as a factor affecting the feed input-sediment quality relation is also demonstrated in the present study.

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Nucleoplasmin, first isolated from Xenopus laevis eggs, promotes nucleosome assembly. Hereby, we have purified a nucleoplasmin-like protein from carp (Cyprinus carpio) eggs using ion exchange and subsequent gel filtration columns. The protein was recognized by a polyclonal antiserum against Xenopus laevis nucleoplasmin and had an amino acid composition similar to other member of the nucleoplasmin family proteins. Partial amino acid sequences from the cyanogen bromide (CNBr)-cleaved fragments showed high homology with Xenopus nucleoplasmin. The protein was also found to form an oligomeric complex and to be phosphorylated. Moreover, this protein promoted sperm nuclear decondensation as well as that of nucleoplasmin from Xenopus laevis eggs. These results suggest that the fish protein isolated here is a member of nucleoplasmin family.

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Taste-active components were determined for a typical first-grade Vietnamese fish sauce Nuoc mam, of which total content of extractive compounds reached as high as 20 g/100 mL. A complete synthetic extract prepared from 35 compounds according to the analytical data reproduced almost satisfactorily the taste of the original fish sauce. From the synthetic extract, 11 compounds were identified to be the taste-active components by a series of omission and addition tests. The components consisted of glutamic and aspartic acids, threonine, alanine, valine, histidine, proline, tyrosine, cystine, methionine, and pyroglutamic acid. The most effective compound for recreating the characteristic flavor of fish sauce was glutamic acid, followed by pyroglutamic acid and alanine. Many of these components contribute to umami, sweetness, and overall taste of fish sauce. Even though the simplified 11-component extract reproduced the taste of fish sauce, the taste strength was weaker than that of the complete synthetic extract or fish sauce itself.

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A Vietnamese fish sauce Nuoc mam used in the present study was rich in oligopeptides of which nitrogen content accounted for 20% of total nitrogen. The high-molecular-weight peptide fractions fractionated by ion-exchange chromatographies and ultrafiltration enhanced sweetness and umami as well as sourness and bitterness of the fish sauce, and increased several flavor characteristics including continuity, first taste and aftertaste. Thus, it is apparent that large amounts of peptides produced during the long-term fermentation of fish sauce are responsible for the complicated taste of the sauce. From each fraction, 17 peptides in total were isolated and determined for their amino acid sequences. These di-, tri- and tetra-peptides synthesized by solid- or liquid-phase method gave any one of bitter, sour, umami taste, or practically no taste in the absence of salt. In the presence of 0.3% NaCl, however, almost all peptides showed sweet and umami tastes. Other than these 17 peptides, there existed many other kinds of peptides in the fish sauce. Thus, these peptides are thought to contribute to the overall taste of the fish sauce.

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Transglutaminase seems to be related to the setting phenomenon of fish meat paste that occurs at temperatures below 40°C. In many reports on the relationship between transglutaminase and setting phenomenon, the enzyme activity has been measured at 25°C. However, it is known that the setting phenomenon is complicated and the effect of calcium and the 3-D structure of myofibrils, which are sensitive to temperature, play important roles in the reaction. In the present study, total activities of transglutaminase of threadfin bream, white croaker, red sea bream and carp meats were measured at various temperatures. Total transglutaminase activities at 25°C of tested fish meats are arranged in order as follows; white croaker, red sea bream, carp, threadfin bream. In contrast, optimal temperature of carp meat is 30°C, red sea bream 40°C, threadfin bream 50°C and white croaker 50-55°C. In carp meat, the enzyme activity at optimal temperature (30°C) became 8.5-fold higher than at 25°C. The data of the total activity at various temperatures are useful in order to comprehend the details of the reaction.

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An ice-nucleating bacterium, designated MACK-4, was isolated from ice-stored mackerel (Scomber australasicus) and identified as a Pseudomonas fluorescens. The optimal temperature and pH for its growth in nutrient broth with 2.5% glycerol (NB-G) were 15°C and 6.5, respectively. The maximal ice-nucleating activity (INA) was obtained after 54 h incubation at 15°C. However, the INA was almost completely lost after 48 h incubation at 25°C or higher. The growth and INA decreased with increase of NaCl added in NB-G within 0.0-4.0%. The INA of MACK-4 was very stable at 5-25°C, pH 4.0-9.5, while that of isolated ice-nucleating matter from MACK-4 was stable at 5-25°C, pH 5.5-9.0.

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