Hidemi Kumai

Status of Bluefin Tuna Farming, Broodstock Management, Breeding and Fingerling Production in Japan

Development of bluefin tuna farming in Japan has a 30-year history. This paper reviews recent developments as well as the current status and problems of farming, broodstock management, and fingerling production in Japan. Farmed bluefin tuna by wild seed-stock comprises approximately 15–20% (2,400 tons) of the annual catch of bluefin in Japan. Comparisons among farms in Japan indicate that bluefin tuna growth is positively correlated with annual water temperature. Broodstock commence spawning at approximately 3–5 years of age, and eggs are collected from within the cages. The number of bluefin tuna eggs collected in Japan has varied greatly—between zero and about 500 million—with variance between farms and years. Current larval rearing techniques produce tens of thousands of fingerlings of 35–50 mm per season in Japan, but survival rates are 0.01–4.5%. Although there are the bottlenecks in egg collection and larval rearing under the present technology, the approaches for enhancing bluefin resources and the replacement of wild seed-stock by artificial fingerling to tuna farms have firmly been improving in Japan.

Growth and morphological development of larval and juvenileepinephelus bruneus (perciformes: Serranidae)

The growth and morphological development of larval and juvenileEpinephelus bruneus were examined in a hatchery-reared series. Average body length (BL) of newly-hatched larvae was 1.99 mm, the larvae growing to an average of 3.96 mm by day 10, 6.97 mm by day 20, 12.8 mm by day 30, 22.1 mm by day 40 and 24.7 mm by day 45 after hatching. Newly-hatched larvae had many mucous cells in the entire body epidermis. By about 4 mm BL, the larvae had developed pigment patterns peculiar to epinepheline fishes, including melanophores on the dorsal part of the gut, on the tips of the second dorsal and pelvic fin spines, and in a cluster on the ventral surface of the tail. Spinelets on the second dorsal and pelvic fin spines, the preopercular angle spine and the supraocular spine, had started to develop by about 6 mm BL. The notochord tip was in the process of flexion in larvae of 6–8 mm BL, by which time major spines, pigments and jaw teeth had started to appear. Fin ray counts had attained the adult complement at 10 mm BL. After larvae reached 17 mm BL, elements of juvenile coloration in the form of more or less densely-pigmented patches started to appear on the body. Squamation started at 20 mm BL. Major head spines had disappeared or became relatively smaller and lost their serrations by 20–25 mm BL.

Production of cloned red sea bream, Pagrus major, by chromosome manipulation

Red sea bream, Pagrus major, is one of the most important fish cultured in Japan. Two clones of red sea bream were produced. Eggs from a mitotic gynogenetic diploid (mitotic-G2N) red sea bream were inseminated either with sperm from a mitotic-G2N male to produce a heterozygous clone (hetero-clone), or with UV-irradiated sperm of Japanese parrot fish (Oplegnathus fasciatus) and the second meiotic division suppressed by cold shock to produce a homozygous clone (homo-clone). Normal diploids were also produced from one male and female as a control. The clonal status of the fish was confirmed by multilocus DNA fingerprinting. The fingerprinting patterns differed between individuals within the normal diploids. However, there was no variation between individuals within hetero- or homo-clones. The patterns of the homo-clones and the mother were identical, and all the bands of homo-clones were also observed in hetero-clones. Thus, the clonal status of homo- and hetero-clones was confirmed and the production of clones from the broodstock of mitotic-G2N was achieved. The hatching rates, survival rates and growth of the hetero- and homo-clones were recorded for a brief comparison with results of diploid controls.

Ontogenetic changes in RNA, DNA and protein contents of laboratory-reared Pacific bluefin tuna Thunnus orientalis

The ontogenetic changes in the growth potential of larval and juvenile laboratoryreared Pacific bluefin tuna were examined based on RNA-DNA and protein-DNA ratios. Experimental fish were reared at the Ohshima Experiment Station of Kinki University Fisheries Laboratory in August 2002. Samples were taken from 13 to 35 days after hatching (DAH). Metamorphosis from larva to the juvenile stage was observed around 23 DAH. Somatic growth of Pacific bluefin tuna was accelerated after metamorphosis. The value of the RNA-DNA ratio from 13 to 19 DAH increased slightly from 3.77±0.58 (mean±SD) to 7.28±2.23. After that, the ratio markedly increased from 13.89±3.71 on 21 DAH to 19.11±4.27 on 23 DAH, which was the end of the metamorphic period. After 25 DAH, the ratio remained at a high level of 15–20. The protein-DNA ratio showed a similar tendency to the RNA-DNA ratio. These results suggest that the rapid increase in the RNA-DNA ratio in the metamorphic period supports the consequent rapid somatic growth in the juvenile stage. The high ratio after the metamorphic period could be because of the species-specific traits large prey exhibit for their survival and because of the tuna’s fast-growth after the juvenile stage.

A possible correlation between environmental chemicals and pigment cell neoplasia in fish

A croaker (Nibea mitsukurii) has a high incidence of the skin melanoma, chromatophoroma, in a Pacific coastal area in Japan. A sea catfish (Plotosus anguillaris) bearing skin melanosis is also found in the same area. For elucidation of a correlation between these pigment cell neoplasms of the skin and environmental contaminants, an epidemiological survey was conducted to determine the distribution and prevalence of tumor-bearing fish. Based upon observations of a high prevalence of skin neoplasms near the discharge point for kraft pulp mills, experiments were conducted to determine the neoplastic induction efficiency of the effluent on the croaker and sea catfish species. Isolation and identification of mutagens in effluent extracts were carried out using the Ames test, followed by mass spectral analysis of mutagenic fractions. The effluent induced a chromatophoroma on one croaker of the 100 tested, and it induced pigment cell hyperplasia on 70 to 100% of the sea catfish. These skin neoplasms were grossly similar to those observed in the field. Five chloroacetones were identified from the Ames-positive fractions of the effluent, and tetrachlorocyclopentene-1,3-dione and two alpha-dicarbonyl compounds were also detected as mutagens. The above experiments indicate that the mutagenic contaminants found in kraft mill effluent may play an important role in the induction of skin neoplastic disease in fish.

Three cone opsin genes and cone cell arrangement in retina of juvenile Pacific bluefin tuna Thunnus orientalis

In bluefin tuna aquaculture, collision of juveniles with the tank or net walls is a major cause of high mortality. This problem may be related to color sensibility of the visual mechanisms of this species. As a first step in understanding of color vision of Pacific bluefin tuna Thunnus orientalis, we applied a molecular technique and histology to study cone cell distribution in the retina of juvenile fish. We isolated three cone opsin genes encoding one blue-sensitive (SWS2) and two green-sensitive (RH2) visual pigments. In situ hybridization revealed that SWS2 mRNA is localized in the single-cone photoreceptors. The localization of the two RH2 signals in distinct cone cells was not determined, probably because of the high homology between the two RH2 genes. Single-cone photoreceptors appeared frequently in the ventral-temporal retina in approximately 80-mm fish and in the temporal retina in approximately 230-mm fish. These cone distributions may define a visual field with best color contrast vision in front and above the fish with a short wavelength (blue) reflecting target (sensed by single cones), and may be enhanced against the longer wavelength (green) background when fish see a target below them (sensed by double cones).

Characterization of transthyretin in the Pacific bluefin tuna, Thunnus orientalis.

Abstract A cDNA encoding transthyretin was cloned from the Pacific bluefin tuna (Thunnus orientalis). This cDNA contains a complete open reading frame encoding 151 amino acid residues. The deduced amino acid sequence is 81% and 55% identical to the gilthead seabream and common carp forms, respectively, and 33–39% to mammalian, reptilian, and amphibian forms. A 1.0-kb transcript was found in the the liver and ovary; the liver is the main source of this protein. Analysis of triiodo-L-thyronine (T3) and L-thyroxine (T4) binding demonstrated that both T3 and T4 bind to bluefin transthyretin. The binding activity of T3 for bluefin transthyretin is higher than that of T4. These results indicate that bluefin transthyretin acts as a transporter of thyroid hormones (THs) in the plasma, and plays an important role in the function of THs in target cells.

Differences in the biochemical content of buoyant and non-buoyant eggs of the Japanese eel, Anguilla japonica

Eggs of the Japanese eel, Anguilla japonica, were obtained by hormonally induced maturation and ovulation, and then categorized as buoyant and non-buoyant types in seawater after insemination. The water content of the buoyant eggs, 93.1%, was significantly higher than that of the non-buoyant eggs, 87.5%. Furthermore, the buoyant eggs had 1.5 times more free amino acids (FAA), 573.5 μmol/g dry weight (DW), than the non-buoyant eggs, 385.8 μmol/g DW, while no significant difference in egg lipid content or protein content was found. Compared to other pelagic eggs reported in the literature, the FAA content of the buoyant eggs was relatively low, although the water content was similar. These results suggest that there is a close relation between the quality of Japanese eel eggs obtained by typical hormonal inducement and the FAA content of the eggs, but the roles of FAA in the acquisition of egg buoyancy and fulfilling the early nutritional requirements are comparatively minor.

Whole‐body heat transfer coefficient and body temperature change of juvenile Pacific bluefin tuna Thunnus orientalis according to growth

Bluefin tuna maintain a higher body temperature than ambient sea water. Body heat is derived mainly from metabolic heat to elevate and maintain regional body temperature that is higher than the ambient, while heat loss is caused by heat transfer throughout the whole body surface and gills. Retention of high body temperature is thought to differ at each growth stage, so that a larger body mass maintains a higher body temperature. We evaluated the whole-body heat transfer coefficient, thermal difference between each tissue and water temperature, and metabolic heat in tissues during swimming of juvenile bluefin tuna as a function of fork length (FL) using a small thermometer and a treadmill-type flow tank. A system for maintaining high body temperature was well developed in fish with FL greater than 20.0 cm. Whole-body heat transfer coefficient was fitted to a −0.695 power of mass. Juvenile bluefin tuna showed a transition speed of 3.0 FL/s at which they switched from aerobic to anaerobic motion.

Testes maturation of reared Pacific bluefin tuna Thunnus orientalis at two-plus years old

Stable reproduction is essential for supplying artificially hatched fish to tuna aquaculture. We observed testes maturation in reared Pacific bluefin tuna (PBT) Thunnus orientalis at 2+ years of age. The incidence of males with mature testes was 25.0%, and 40% of the males had developing testes that contain spermatozoa, while oocytes of the same aged females were not mature. These fish were wild-caught at 0+ years old in August 1997 and the gonads were examined in October 1998 and January–February 2000. Therefore, the age at examination in 2000 was estimated to be 2 years and 7–10 months old considering the spawning season of the wild PBT and the size when captured. Histological examination of thematured and developing testes showed that they contained spermatozoa, spermatids, spermatocytes, and spermatogonia. All the spermatozoa were observed to be motile in sea water under light microscopy. From the results of this and previous studies, matured males are probably fertile for at least 5 months a year in Kushimoto. The testes maturation observed at young age in captivity is considered promising to reduce the cost of brood-stock maintenance for the juvenile production of PBT, especially if the sperm are cryopreserved.