Recent advances in the functional morphology of chloride cells in killifish Fundulus heteroclitus, a euryhaline teleost with seawater (SW) preference, were reviewed. Immunocytochemical detection of chloride cells with anti-Na⁺/K⁺-ATPase revealed transitional processes of the chloride cell distribution during early life stages. Chloride cells first appear in the yolk-sac membrane at an early embryonic stage, followed by their appearance in the body skin in the later embryonic stages. The principal site for the chloride cell distribution then shifts from the yolk-sac membrane and body skin during embryonic stages to the gills and opercular membrane in larval and later developmental stages. Morphologically distinct SW- and freshwater (FW)-type chloride cells were identified in adult killifish adapted to SW and FW, respectively. Both types of chloride cells are equally active in the two environments, but exert different ion-transporting functions. Following direct transfer of killifish from SW to FW, SW-type chloride cells were transformed into FW-type cells as a short-term response, followed by the promotion of chloride cell replacement as a long-term response. In killifish acclimated to low NaCl (0.1 mM) FW, intense immunoreactivity for vacuolar-type proton pump (V-ATPase) was detected in the basolateral membrane of FW-type chloride cells, whereas the immunoreactivity was much weaker in fish acclimated to FW with higher NaCl concentrations (1.0 and 10.0 mM). These results suggest the occurrence of active ion absorption in FW-type chloride cells and the involvement of V-ATPase in ion-absorbing mechanisms. In view of recent advances in this field, future chloride cell research should be considered in relation to the functional diversity of chloride cells.

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The reproductive traits and the monthly larval abundance of the mantis shrimp Oratosquilla oratoria were investigated in Tokyo Bay, Japan, in 2002. The goal of the study was to elucidate the cause of changes in the monthly pattern of larval abundance from the 1980s to the 1990s as these changes relate to variation in the stock size of the adult shrimp. Oogenesis was divided into 10 stages by histological observation. The developmental stage of oocytes in an individual's ovary was synchronous, suggesting that almost all the oocytes in an ovary are spawned at the same time. The size at first maturity was estimated to be 7≤body length (BL)<8 cm. Fecundity was expressed as a function of BL, ranging from 19 300 eggs for 8 cm BL to 92 100 eggs for 14 cm BL. Small female shrimps (<10 cm BL) spawned around August. Most large female shrimps (≥10 cm BL) spawned around May, and some large female shrimps also spawned until September. Although most large female shrimps spawned in spring, the larval abundance was low before July and high from August onwards. The results suggest that a substantial decrease in the stock size of large individuals causes the low larval abundance before July.

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Kawakawa Euthynnus affinis is the most abundant species among the small tuna in the coastal waters of Taiwan. The migration pattern of kawakawa to Taiwan was determined in the present study based on the species' feeding habits and reproductive biology and on temporal variation of catch for kawakawa and its prey fishes. Schools of this species successively migrate to Taiwan from September to May, and then move away in June. They travel to the waters off Taiwan with the warm Kuroshio current, migrate toward coastal waters for feeding and move away for spawning. It is suggested in the present study that the kawakawa spawning ground is in the waters of the northern Philippines. The migration of kawakawa exhibits size-segregating behavior. Schools of immature young fish first arrive in the coastal waters around Taiwan, followed by the adults.

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Morphometric differences were used to elucidate the stock structure of sword prawn Parapenaeopsis hardwickii in the East China Sea and Taiwan Strait. Six samples were collected separately from the north of the East China Sea (NECS-02) and waters off Tamsui (Tamsui-10), Taichung (Taichung-11 and Taichung-02), Putai (Putai-02) and Cheding (Cheding-02). Eleven measurements made for each individual were size-standardized by allometric methods and the resulting measurements were analyzed by cluster analysis and canonical variate analysis (CVA). Randomization tests were used to test the morphometric variation between groups derived from cluster analysis and CVA. The results of cluster analysis and CVA indicated that the six samples were clustered into three groups: the first group included the NECS-02 sample; the second included the Tamsui-10 and Taichung-11 samples; and the third included the Taichung-02, Putai-02 and Cheding-02 samples. Randomization tests showed that morphometric differences among the three groups were significant. Animals with the same morphometric measurements are often assumed to constitute a stock, so there appear to be at least three morphologically distinguishable stocks of this species in the East China Sea and Taiwan Strait. The first stock is in the north of the East China Sea, the second is in the waters adjacent to Tamsui and Taichung, and the third is in the waters adjacent to Cheding, Putai and Taichung.

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Antimicrobial peptides serve as an important component of the innate immune system of all species by functioning to provide a rapid first line defense against infection. Arthropod antimicrobial peptides have been well described in insects, whereas only a few molecules have been identified in crustaceans. Five variants (types 1-5) of Marsupenaeus japonicus crustin-like peptide cDNA that were obtained from a hemocyte cDNA library and polymerase chain reaction (PCR) amplification are reported here. Marsupenaeus japonicus crustin-like peptide type 1, the predominant type, has a cDNA consisting of 679 nucleotides and an open reading frame consisting of 573 base pairs coding for 191 amino acid residues. Other types contain varying glycine-rich repeats at the N-terminal amino acid sequences. The deduced amino acid sequences of these variants are highly similar to those of Litopenaeus setiferus (80% identity), Litopenaeus vannamei (80% identity) and Carcinus maenas crustins (44% identity). Expression of Marsupenaeus japonicus crustin-like peptide mRNA was detected in hemocytes, but not in the heart, hepatopancreas, gill, fore-gut, mid-gut, muscle, subcuticular epithelium or ovary. The expression level of crustin-like peptide mRNA increased significantly 1, 3 and 7 days post-peptidoglycan feeding as determined by quantitative real-time PCR. These results suggest that crustin-like peptide could have an important role in shrimp defense mechanisms.

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The spawning bed selection of herring Clupea pallasii off Minedomari, Atsuta, on the west coast of Hokkaido, Japan, is discussed in relation to the topography of the rocky shore and wave conditions during the spawning season. From 1998 to 2003, herring have spawned their eggs mainly on the leaves of the seagrass Phyllospadix iwatensis Makino in almost the same site on the rocky shore off Minedomari in the Atsuta area. The site is connected to a valley-like feature offshore. Wave conditions were surveyed at Minedomari during the spawning season in 2000 and the wave height was estimated from 1998 to 2003, except for in 2000, using the correlation of wave height between Minedomari and Ishikari Bay New Port, approximately 18 km south-west of Minedomari. Herring spawned under calm conditions, during which the wave height was approximately 0.5 m off Atusta and Aoshima and was 0.18-1.28 m off Minedomari. The distribution of water particle velocity on the sea bottom surface as a result of ocean waves off the Minedomari area, which was estimated based on the wave height and the topography of the coast, suggested herring could swim easily into the shallower area along the valley-like feature off Minedomari. Therefore, topographical features are thought to be one of the reasons why herring have used Minedomari as a spawning bed. Additionally, seepage of freshwater from the bottom, which was observed in this area, could also be the reason why herring spawn in Minedomari repeatedly.

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In order to find out the role of sodium bicarbonate (NaHCO₃) on the initiation of sperm motility in the Japanese eel Anguilla japonica, interactions were investigated between NaHCO₃ and various reagents (K⁺ channel blocker 4-aminopyridine [4-AP], ammonium chloride [NH₄Cl], sodium acetate and calcium chloride [CaCl₂]) that could regulate internal factors (intracellular K⁺, intracellular pH [[pH]_i] and intracellular Ca²⁺) in sperm motility. Contradictory effects of NaHCO₃ were observed (i.e. an inhibitory effect when 4-AP was absent and a promoting effect when 4-AP was present). Sodium bicarbonate inhibited the initiation of sperm motility in the Japanese eel. However, NaHCO₃ restored the motility of immotile sperm that 4-AP inhibited. The inhibitory effect of NaHCO₃ disappeared with the addition of NH₄Cl, which raised [pH]_i, but the promoting effect was not affected by [pH]_i. Although NaHCO₃ recovered motility in the presence of 4-AP, this recovery was also observed with the addition of CaCl₂ instead of NaHCO₃. In the initiation of sperm motility in the Japanese eel, two roles for NaHCO₃ are suggested: an inhibitory role relating to the regulation of [pH]_i and a promoting role relating to the uptake of another initiation factor, which could be Ca²⁺.

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To clarify faunal overlap between a seagrass bed and adjacent coral and sand areas, and the number of reef fishes utilizing the seagrass bed as juvenile habitat, visual censuses were conducted at Amitori Bay, Iriomote Island, Japan. The numbers of species and individuals of fishes were significantly higher in the coral area than in the seagrass bed and sand area. Cluster and ordination analyses based on the number of individuals of each species demonstrated that the fish assemblage structure differed among the three habitats in each season, but with some overlaps. Approximately half the seagrass bed fishes occurred in the adjacent coral area (coral-seagrass species). Dominant species of coral-seagrass species utilized the seagrass bed as an important juvenile habitat. Thus, some overlaps in habitat use were present between the seagrass bed and adjacent coral area. Despite such overlaps, however, coral-seagrass species accounted for only approximately 15% of coral reef fishes overall, indicating that most of the latter hardly utilize the seagrass bed directly in the study area.

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Presence-absence sampling (PAS) is a sampling technique that observes only the presence or absence of objects, such as eggs, in sampling units. The purpose of the present paper was to study the oversight probability in a PAS study for detecting fish eggs in connection with the spatial distribution of eggs. It was shown that the oversight probability of presence of eggs under no-eggs observation is a monotone decreasing function of the parameter of the degree of contagion.

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A pleuronectiform fish, the barfin flounder Verasper moseri, expresses three gonadotropin-releasing hormone (GnRH) forms in the brain: salmon GnRH (sGnRH), chicken GnRH-ll (cGnRH-ll) and seabream GnRH (sbGnRH). To clarify the effects of photoperiod on GnRH systems, changes in brain and pituitary GnRH peptide levels were examined using time-resolved fluoroimmunoassays. In experiment 1, 5-month-old male barfin flounder (mean total length 9.0 cm, body weight 11.0 g) were divided into short (8:16 h light: dark [L:D] cycle; lights on 08.00-16.00 hours) and long photoperiod (16:8 h L:D cycle; lights on 04.00-20.00 hours) groups in mid September and maintained until November under natural water temperature (19.3-15.2°C). Brain sGnRH concentrations were significantly higher in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in total length, body weight, plasma testosterone concentration, brain cGnRH-ll concentration and pituitary sbGnRH content. In experiment 2, 7-month-old male barfin flounder (mean total length 16.5 cm, body weight 76.8 g) were divided into short and long photoperiod groups in mid December and maintained until February under natural water temperature (12.5-6.6°C). Total length, body weight and condition factor were significantly greater in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in plasma testosterone concentration and GnRH levels in the brain and pituitary. These results indicate that levels of sGnRH in barfin flounder are influenced by photoperiodic treatment dependent on water temperature and/or body size.

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Black porgy are hatched and released in marine ranching areas in Japan, Korea and Taiwan to maintain fisheries resources in coastal areas. If acoustic methods are used for field surveys, target strength (TS) information on the species is needed to convert acoustic data to biomass density. Target strength of black porgy (Acanthopagrus schlegeli) 15.5-32.9 cm fork length (nine individuals) was measured at 38 and 120 kHz (split beam) by ex situ measurements (cage and fishhook methods) of living fish, a tethered method with shock-frozen fish, and an acoustic scattering model, the Kirchhoff ray mode (KRM) model based on fish morphology. The ex situ TS-to-fork length (FL, cm) relationships were TS_38kHz=20log₁₀FL-64.6(r=0.77) and TS_120kHz=20log₁₀FL-65.2(r=0.70). The values for the KRM model were TS_38kHz=20log₁₀FL-64.8(r=0.88) and TS_120kHz=20log₁₀FL-65.4(r=0.80). The agreement between the ex situ measurements and the KRM model for the mean TS was very good. In contrast, the results for the tethered method (TS_38kHz=20log₁₀FL-69.3[r=0.42]) differed markedly from the other two methods, perhaps because of the effects of the shock-freezing procedure in air. The results of the ex situ measurements and scattering model provide basic information for studying TS and conducting acoustic surveys of black porgy, even though the tethered method must be modified.

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Age-based population assessment is widely used, but there are cases where information on age or even body length of landed fish is difficult to collect. In the present study, the biomass and fishing mortality of the southern Hokkaido stock of Pacific cod Gadus macrocephalus was estimated directly from body weight composition, using weight-based virtual population analysis, from 1994 to 2000. The estimated biomass over 1 kg body weight was 5607 t in 1994 and increased to 7908 t in 2000. The increase was explained by an increase in recruitment.

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This study is the first to report on the high occurrence of agglutinated pelvic fin membrane deformities in hatchery-reared black rockfish Sebastes inermis. For 5 years, this symptom was marked in hatchery-reared fish, with 58.7% of fish deformed on average (varying between 46.7 and 72.0%). The deformity was a peculiarity in hatchery-reared fish, but is negated in wild fish, and was not related to whether the fish broodstock originated from the wild or from a hatchery. Mark-release experiments showed that deformed fish were almost the same as normal hatchery-reared fish in growth and survival rates, and, theoretically, the recapture numbers of hatchery-stocked fish, estimated by deformity, almost coincided with actual recapture numbers, confirmed by otolith tagging. The results of the present study indicate that deformity in hatchery-reared black rockfish is useful as a stock separation tool.

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Age and growth of alfonsino Beryx splendens (Lowe) were studied using counts of presumed daily incremental growth rings on the transverse section of otoliths of fish collected from the Kanto District, central Japan. Microstructural growth increments were observed from the core to the outermost margin of the broadest of approximately 50 branches formed on the surface of an otolith. Of 98 otoliths examined, 46 were readable. The number of increments and fish lengths ranged from 448 (218 mm fork length [FL]) to 3701 (411 mm FL). The ages of these fish were estimated to be 1 year, 2 months and 10 years, 2 months, respectively, assuming that an increment was formed daily. The von Bertalanffy growth equation combined for males and females was expressed as follows: L_t=542{1-exp[-0.133(t+2.00)]}, where L_t is fork length (mm) at yearly age t. The results of the present study were compared with those of other researchers who had studied alfonsino from central Japan.

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This study was carried out to investigate the genetic variability and the population genetics of Sparus auratus. In fact, despite its importance in Mediterranean fisheries and aquaculture, very little it is known concerning its population structure. Samples of wild gilthead sea bream were collected in seven different localities along the Mediterranean and Atlantic coasts and were genetically characterized by means of microsatellite markers. Genotyping at four microsatellite loci revealed high polymorphism (7-38 alleles/locus) and expected heterozygosities, which ranged from 0.80 to 0.85. A slight but significant population structure was found (F_ST=0.010). In fact, at least three populations of gilthead sea bream within the Western Mediterranean Sea were identified (Sardinian Sea, Sardinian Channel and Central Tyrrhenian Sea), which are also genetically differentiated from those of the Atlantic Ocean and the Adriatic Sea.

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In the present study, rubber bands were tied around the girth of rainbow trout Oncorhynchus mykiss to simulate and assess the physiological damage and survival rates of fish that encounter gill nets. Physiological condition was assessed by analysis of blood lactate, pH, oxygen partial pressure and plasma potassium concentration before and after release from binding. Twenty-four hours after release, half of the test fish had died. Although the binding forces used to hamper fish did not differ significantly, blood lactate levels in the dead fish rose to 10-fold higher than those in the survivors. Consequently, the pH level fell in the group that died, whereas in the surviving group it fell only slightly and soon recovered. Potassium concentration increased after release from binding in fish that died, and the PO₂ levels in these fish continued to drop until just before death. It was concluded that when fish get entangled in fishing nets, blood lactate accumulates because of physical fatigue and the elevated lactate levels lead to metabolic acidosis. Finally, fatal metabolic conditions could cause higher mortality rates, even in escapees.

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Small abalone meats were heated at 80°C and 98°C for 0-120 min and the differences in chemical, physical and sensory changes of the cooked meats were investigated. The decrease in moisture and weight and the increase in browning and Hunter's b-value were relatively higher for cooking at 98°C than at 80°C. After cooking for 20-120 min, the total amount of adenosine triphosphate and its related compounds on a dry weight basis decreased by 17-27% at 80°C and by 30-39% at 98°C; the total amount of free amino acids on a dry weight basis changed insignificantly at 80°C but decreased by 22-35% after cooking at 98°C. The meats cooked at 80°C were higher in cutting force whereas the levels in the samples cooked at 98°C did not decrease until samples had been cooked for 60 min. The hydroxyproline content showed little change during cooking except for in samples cooked at 98°C for 120 min, in which the content was found to be low. The extended cooking at 80°C improved the acceptability of small abalone meat, whereas only the acceptability score of aroma increased significantly for cooking at 98°C.

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In the present study, the cDNA encoding myoglobin (Mb) of bigeye tuna Thunnus obesus was cloned and its amino acid sequence deduced in order to investigate the relationship between the primary structure and thermostability of scombridae fish Mb. An open reading frame of bigeye tuna Mb cDNA contained 444 nucleotides encoding 147 amino acids. The primary structure of bigeye tuna Mb was highly conserved when compared with those of bluefin tuna and yellowfin tuna Mb, the sequence identity being 95.2-100.0%. It also showed relatively high identity (82.3-89.1%) with the counterparts of scombridae fish. Myoglobin was then isolated from the dark muscle of four scombridae fish including bigeye tuna. Differential scanning calorimetry and circular dichroism measurements on these Mb revealed that the thermostability of bigeye tuna Mb was lowest and that of skipjack Katsuwonus pelamis Mb highest among the scombridae fish Mb examined. The α-helical contents of scombridae fish Mb at 10°C were in the range of 39.8-44.8%, clearly lower than that of horse Mb (55.3%), suggesting instability of fish Mb. The melting temperatures of these Mb fell in the range of 75.7-79.9°C, lower than that of horse Mb (84.2°C). These results strongly suggest the instability of fish Mb.

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The complete mitochondrial genome sequence of 10 walleye pollocks, Theragra chalcogramma, from the Japan Sea and Bering Sea was determined. The 16 568-16 571 bp genome contains the same 37 mitochondrial structural genes (two ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes) as found in all other vertebrates analyzed, in an organization identical to that of other bony fish. The major non-coding region had several conserved sequence features. Nucleotide variations of ND1, ND5, and control region were high, and these regions appear to be good candidates for high-resolution markers in population studies.

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Carp myosin was conjugated with alginate oligosaccharide (AO) through the Maillard reaction under low relative humidity, and the functional properties of the myosin-AO conjugate were investigated to clarify the role of myosin in the functional improvement of fish myofibrillar proteins (Mf) by the glycosylation. The findings were as follows. First, myosin became highly solubilized at lower NaCl concentrations by conjugation with AO and NaCl-dependence of the solubility was lost when>12% of the available lysine residues were reacted with AO and 50 μg/mg of AO was attached to myosin. Second, the thermal stability of myosin was effectively improved by conjugation with AO. Heat-treatment at 50°C for 6 h has no effect on the solubility of the myosin-AO conjugate regardless of the NaCl concentration. Third, the improved functionalities of myosin conjugated with AO remained even at a nearly isoelectric point. The improving effect of AO-conjugation on the characteristics of myosin was almost the same as Mf reacted with AO. Therefore, it is apparent that that improved functionalities of the glycosylated Mf reflect the functional changes of myosin.

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Lipid class and fatty acid composition of phospholipids from the gonads of skipjack tuna were examined to evaluate effective utilization of the processing of by-products. The predominant phospholipids in the ovaries were phosphatidylcholine (PC; 47.9%), phosphatidylethanolamine (PE; 19.3%) and lyso-phosphatidylcholine (LPC; 19.1%). In contrast, those in the testes were PC (40.1%), PE (29.3%) and phosphatidylserine (PS; 9.6%). The percentage of docosahexaenoic acid (DHA) was markedly high at more than 50% in LPC of the ovaries, and PE and PS of the testes were also high. The percentages of DHA at sn-position 2 of the predominant phospholipids, except for PC in the testes, were more than 60%, in particular PE in the testes was remarkably high at 81.9%. After storage for 2 days at 5°C, the LPC content in the ovaries increased twofold and the DHA level of LPC was the same as before, though the contents of other phospholipids decreased.

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The photoprotective activity of the scallop shell water-extract was investigated using cultured rat skin keratinocyte cells. It was found that the scallop shell water-extract protected keratinocyte cells against ultraviolet (UV)-B-induced damage. The scallop shell water-extract had protective effects against hydrogen peroxide (H₂O₂) in skin keratinocyte cells and also showed 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity and lipid peroxidation inhibitory activity. In addition, the scallop shell water-extract promoted proliferation of keratinocyte cells. The scallop shell water-extract may protect skin keratinocyte cells against UV-B-induced damage through two factors; antioxidant activity and growth-promoting activity in skin keratinocyte cells. These results suggest utilization of the scallop shell water-extract as materials for protecting skin.

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The two Staphylococcus strains that had been isolated from fish sauce mush (moromi) made from frigate mackerel in Japan and proved to improve fish sauce odor, were examined for their taxonomic positions. The sequence analysis based on 16S rRNA and rpoB showed that the two strains, R4Nu and R5G, had an identical sequence with sequence identities of 99.5% and 99.0% to the above two genes from the closest species of S. nepalensis, respectively. A DNA hybridization test of the two strains showed more than 80% DNA similarity with S. nepalensis, thus confirming the above-mentioned species identification. Polymerase chain reaction primers specific to the strain isolated from fish sauce mush were designed from rpoB and examined for the distribution of this species to various fish sauces made in Asian countries as well as to fish sauce starter (malt) made from soy beans and barley in Toyama Prefecture, Japan. The amplified DNA fragment bearing the S. nepalensis gene was detected in the enriched culture of the malt, although no positive reaction was shown with fish sauce samples. These results suggest that S. nepalensis indebted to improve fish sauce odor was originated from the fish sauce starter malt.

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