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Female fishes allocate a certain quantity of lipids for egg production once mature. In aquaculture, such physiological changes often result in the deterioration of flesh quality. In the study reported here, we aimed to characterize the seasonal changes in the lipid contents in aquaculture-produced female Pacific bluefin tuna (PBT) Thunnus orientalis, which is one of the most commercially important species for aquaculture worldwide. We analyzed the seasonal changes in the lipid content and weight of tissues and organs responsible for lipid allocation towards egg production, including the muscles, liver, mesenteric fat (perigonadal fat), and ovaries. During the spawning season, the total lipid content of the muscles and liver of the mature females decreased, whereas that of the ovaries increased; similarly, the weight of perigonadal fat decreased. We hypothesized that muscle, liver, and perigonadal fats contribute significantly to the accumulation of lipids during the maturation of female PBT. This is the first study to report an association between a reduction in muscle lipid content and sexual maturation in aquaculture-produced female PBT. These findings provide important insights regarding efficient aquaculture operations for this economically important fish. In addition, this study is the first to provide data on the association between the lipid dynamics of female PBT and sexual maturation in aquaculture, further enhancing our understanding of the energy allocation strategy during maturation of female bluefin tuna. [ABSTRACT FROM AUTHOR]

Hatchery-produced Pacific bluefin tuna (PBT) Thunnus orientalis juveniles experienced mass mortality after being transferred to sea cages in a cove on Kakeromajima Island, Kagoshima, Japan, in July 2022. Manca and juveniles of cymothoid isopods were highly prevalent in the branchial cavity of the few surviving PBT juveniles. We identified two species of cymothoid parasites based on their mitochondrial DNA sequences. Most parasites had cytochrome c oxidase subunit 1 (COI) and 16S rDNA sequences similar to those of Norileca indica, while some were identified as Ceratothoa carinata. Adult cymothoids were found in the bigeye scad Selar crumenophthalmus in the same area and were also morphologically and molecularly identified as N. indica. Therefore, it is likely that the mancae and juveniles of the cymothoids accidentally infested the PBT juveniles after they were released from adult parasites in wild natural hosts, such as S. crumenophthalmus, living around the PBT farming cages. This is the first record of N. indica in Japan and the northernmost record of this species. To our knowledge, this is the first report of mass mortality in PBT juveniles caused by parasitic isopods. Our study is the first step in developing measures to avoid the damage caused by the parasites. [ABSTRACT FROM AUTHOR]

A variety of tunas of the genus Thunnus are consumed daily in Japan as sliced raw fish (sashimi and sushi). The consumption of fresh sliced raw fish, i.e., unfrozen or uncooked, can sometimes cause food poisoning that is manifested by transient diarrhea and vomiting for a single day. One of the causes of this type of food poisoning has been identified as live Kudoa septempunctata (Myxosporea: Multivalvulida) in the olive flounder ( Paralichthys olivaceus). Furthermore, raw slices of fresh tunas are highly suspected to be a possible causative fish of similar food poisoning in Japan. In the present study, we conducted a survey of kudoid infections in tunas (the yellowfin tuna Thunnus albacares, the Pacific bluefin tuna Thunnus orientalis, and the longtail tuna Thunnus tonggol) fished in the western Pacific Ocean off Japan and several East Asian countries and characterized morphologically and genetically the kudoid myxospores in pseudocysts or cysts dispersed in the trunk muscles. Pseudocysts of solely Kudoa hexapunctata were identified in the Pacific bluefin tuna (four isolates), whereas in the yellowfin tuna (21 isolates) pseudocysts of Kudoa neothunni and K. hexapunctata were detected at a ratio of 15:6, respectively, in addition to cyst-forming Kudoa thunni in five yellowfin tunas. In the trunk muscles of six longtail tunas examined, pseudocysts of K. neothunni (all six fish) and K. hexapunctata (two fish) were densely dispersed. The myxospores of K. neothunni found in these longtail tunas had seven shell valves and polar capsules (SV/PC) instead of the more common six SV/PC arranged symmetrically. Nucleotide sequences of the 18S and 28S ribosomal RNA gene (rDNA), some with the internal transcribed spacer regions as well, of K. hexapunctata and K. neothunni from the three Thunnus spp., including the seven-SV/PC morphotype, were very similar to previously characterized nucleotide sequences of each species, whereas the 18S and 28S rDNA of four isolates of K. thunni from yellowfin tunas showed a range of nucleotide variations of 99.0-99.9% identity over 1752-1763-bp long partial 18S rDNA and 97.4-99.9% identity over 797-802-bp long partial 28S rDNA. Therefore, this rather high variation of the rDNA nucleotide sequences of K. thunni proved to be contrary to the few variations of K. neothunni and K. hexapunctata rDNA nucleotide sequences. The present study provides a new host record of the longtail tuna for K. neothunni and K. hexapunctata and reveals a high prevalence of the seven-SV/PC myxospore morphotype of K. neothunni in this tuna host. [ABSTRACT FROM AUTHOR]

The genetic population structure of the Pacific bluefin tuna (PBF) Thunnus orientalis and the yellowfin tuna (YFT) T. albacares in the North Pacific Ocean was investigated. The polymorphism of microsatellite (SSR) loci and sequences of mitochondrial DNA control region (mtCR) were analyzed for 71 samples of PBF from Japan and Mexico and 45 samples of YFT from Japan and Panama. In the SSR analyses, both single-locus (−0.010 to 0.008 in PBF and −0.023 to 0.020 in YFT) and global multilocus (0.003 in PBF and −0.002 in YFT) F values among the geographic populations were low and not significant in these species. In the mtCR analyses, neither the neighbor-joining tree nor the minimum spanning network showed genetic differentiation among the geographic populations in each species. The pairwise F values among the geographic populations of them (−0.005 in PBF and −0.020 to −0.014 in YFT) were low and not significant. Our SSR and mtCR data suggested that genetic differentiations were not evident among the eastern and western populations in the North Pacific Ocean either in PBF or in YFT. Mismatch distributions, demographic parameters, and neutrality tests suggested that sudden population expansion of PBF and YFT in the North Pacific Ocean occurred 628,000-731,000 and 450,000-525,000 years ago, respectively. [ABSTRACT FROM AUTHOR]

A feeding experiment was conducted to evaluate the efficacy of, and determine the requirements for dietary AsA ( l-ascorbic acid) using its popular derivative l-ascorbyl-2-monophosphate-Mg (APM) for juvenile Pacific bluefin tuna (PBT). The five test diets (with APM-0, -400, -800, -1200 and -1600) were prepared by adding 0, 400, 800, 1200 and 1600 mg of APM kg diet, respectively. These diets, together with chopped sand lance (the control diet), were administered, in duplicate to 0.27 g PBT at 25 days after hatching (300 juveniles per 15 m tank). The PBT were fed to apparent satiety 6 times a day for 2 weeks. While final body weight, feed conversion and survival showed no differences among diets APM 400-1600, PBT fed APM-0 indicated lower growth performance together with anorexia, dark pigmentation and ataxia from 5 days and reached 80 % mortality at 12 days after the start of the feeding trial. In addition, liver and brain ascorbic acid (AsA) concentrations increased with increasing dietary APM levels, the values were similar in PBT fed diets with APM-1200 and APM-1600, and liver and brain concentrations of AsA were significantly higher in these fish than in the PBT fed the other diets ( P < 0.05). The PBT responded to increasing dietary APM, and the fish fed APM-1200 had the highest weight gain, protein and lipid content. The data indicate that PBT can utilize APM as an AsA source and suggest that dietary supplementation with APM-1200 (454 mg AsA kg diet) would result in satisfactory growth. [ABSTRACT FROM AUTHOR]

During the parasitological survey of cultured juvenile Pacific bluefin tuna (PBT) Thunnus orientalis in 2007 and 2008, two myxosporeans and one microsporidian were found. Morphological and molecular analysis showed that the heart-infecting and brain-infecting myxosporeans are identified as Kudoa shiomitsui and K. yasunagai, respectively. This is a new host record for both species. High prevalence of infection (77-100%) with K. shiomitsui was observed in October to December (1-2 months post-transfer to sea cages), whereas only a few fish were infected with K. yasunagai. A microsporidian observed as white 'cysts' in the trunk muscle of PBT had a resemblance to Microsporidium seriolae, the causative parasite of beko disease in yellowtail. Phylogenetic analysis showed that the microsporidian from PBT is closely related to but distinct from several other muscle-infecting species such as M. seriolae, Microsporidium sp. RSB, and Microsporidium sp. SH. Additionally, the spore dimension (2.7 × 1.5 μm on average) was remarkably smaller than the others. These results suggest that the present microsporidian is an undescribed species and designated Microsporidium sp. PBT. Prevalence and intensity of infection with Microsporidium sp. PBT reached a maximum of 100% and 20 cysts/fish, respectively. Although pathogenic effects of the two Kudoa species on fish health remain unknown, the microsporidian could be of concern to PBT aquaculture due to unsightly cysts in the musculature, reducing the market value of the fish. [ABSTRACT FROM AUTHOR]

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). [ABSTRACT FROM AUTHOR]

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 the matured 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 broodstock maintenance for the juvenile production of PBT, especially if the sperm are cryopreserved. [ABSTRACT FROM AUTHOR]

The digestive enzyme activities of Pacific bluefin tuna Thunnus orientalis were evaluated for specific activity and characterized for pH and temperature optima in crude extracts of stomach, caecal mass, and proximal, middle and distal intestine. A higher level of alkaline proteolytic activity was detected in the caecal mass than in the proximal intestine. Total alkaline proteases, trypsin, chymotrypsin and leucine aminopeptidase (LAP) were tested. The temperature and pH analyses showed that proteolytic activity as well as lipase were maximal in the alkaline range, with a maximum at pH 9.0 and at temperatures between 35 and 60°C, except for the pepsin, which showed maximum activity at the same temperatures but in the acid range (pH 3.0). The α-amylase activity showed a broader range in activity, both for pH and temperature, with higher activity over the alkaline pH values and higher temperature. The lipase activity seems to be nondependent on bile salts under our assay conditions, resulting in a significant activity reduction in the presence of bile salts. This knowledge will allow the development of a gastrointestinal model (everted intestine) where food or feed will be hydrolysed with the fish’s own enzymes, a project that is being undertaken in our laboratory as a contribution to the development of novel diets for tuna fish. [ABSTRACT FROM AUTHOR]

Pacific bluefin is a highly valuable pelagic species that inhabits a broad range in the North Pacific Ocean. The reproductive biology, especially for the spawning aggregation in the south-western North Pacific Ocean, is not well understood. Thus, a total of 119 paired ovary specimens were collected from the Taiwanese longline fleet during the 1999 fishing season (late April through June) to gain a better understanding of important reproductivity-related stock parameters associated with this species. The following conclusions were made: (i) condition factor decreased from late May to early June; (ii) the sex ratio might be 1:1 for spawners; (iii) the gonadosomatic index stayed at a relatively high level and markedly increased from late May to early June; (iv) histological examination of oocytes indicated that all specimens were sexually mature; (v) spawning activity appeared to start in May and peak in late May to early June; (vi) batch fecundity increased with fork length; and (vii) preliminary estimates of spawning frequency between batches ranged 2-4.5 days based on analysis of postovulatory follicles. [ABSTRACT FROM AUTHOR]

Using the skeletal muscle of full-cycle cultured Pacific bluefin tuna (body weight: 13.1 ± 2.6 kg, cultured for about 21 months), the proximate compositions, breaking strength and histological structure of the front and rear parts of the dorsal ordinary muscles (FD-OM and RD-OM) and the ventral ordinary muscles (FV-OM and RV-OM) were compared. The FV-OM showed low moisture, protein, ash and high fat contents ( P < 0.05, respectively) for the other three positions. The breaking strength of FD-OM, RD-OM and RV-OM increased up to 15–18 h and decreased later. However, the breaking strength of FV-OM was maintained during chilled storage. The pH of all muscles decreased up to 15 h, and then stayed at pH 5.7–5.8. However, the pH of FV-OM stayed at a higher level (pH 5.9). The histological structure observed by optical microscopy showed a space extension among muscle cells after 24 h in all positions, and these results were also supported by image analysis. [ABSTRACT FROM AUTHOR]

The pacific bluefin tuna (PBT) Thunnus orientalis is one of the most important species for fisheries in the world. This species has been highly exploited in fisheries, resulting in dwindling of its natural resources. The artificial hatching of a third generation of fully cultured PBT was a success in 2007, suggesting the possibility of mass production of PBT for food and for seed release. To enhance stock structure studies and investigate genetic feature of seedlings, we isolated 25 polymorphic microsatellite loci (2-18 alleles/locus; expected heterozygosity, 0.042-0.941) from PBT. [ABSTRACT FROM AUTHOR]