Changes of proximate compositions and myoglobin content in the dorsal ordinary muscles of the cultured Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> with growth

ABSTRACT:   Using the dorsal ordinary muscle (DOM) of cultured Pacific bluefin tuna (body length [BL]: 47.5–81.8 cm, body weight [BW]: 2.1–13.5 kg, n  = 15), the changes of proximate compositions and myoglobin (Mb) content with growth were investigated. There was a positive correlation ( r  = 0.9832, P  < 0.05) of BL and BW in cultured tuna. The protein contents of the DOM of cultured tuna decreased ( P  < 0.05) and the lipid contents had a tendency to increase (not significantly) with growth. The meat color changed from pink to red with growth. In addition, the Mb contents of the DOM of cultured tuna increased ( P  < 0.05) from 1.0 mg/g (minimum BW fish) to 3.8 mg/g (maximum BW fish) with growth. These results indicate that the increase of the Mb content in the DOM of cultured tuna is not caused by the restriction of exercise and overfeeding between 2.1 kg and 13.5 kg of BW.     

Changes in myoglobin content and proximate compositions of the dorsal ordinary muscles of full-cycle cultured Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) with body size

We investigated the changes in myoglobin (Mb) content and proximate compositions of the cephalad parts of the dorsal ordinary muscles (Ce‐DOM) of full‐cycle cultured (FC) Pacific bluefin tuna [body length: 42.6–115.4 cm, body weight (BW): 1.7–38.4 kg, killed from July 2004 to October 2005]. The Mb content of Ce‐DOM increased (P<0.01) until a BW of 7.0 kg was reached. However, the Mb content of Ce‐DOM had a wide variety above a BW of 7.0 kg. On the other hand, the moisture, protein and ash contents of Ce‐DOM of FC tuna decreased (P<0.05 and 0.01) with an increase in BW. However, the lipid content of Ce‐DOM increased gradually above about a BW of 20.0 kg. Furthermore, the Mb content in protein of the Ce‐DOM of FC tuna increased with an increase in BW. These results indicated that the increase in Mb content of Ce‐DOM of FC tuna was not related to changes in proximate composition between a BW of 1.7 and 38.4 kg.     

Effect of fasting on physical/chemical properties of ordinary muscles in full-cycle cultured Pacific bluefin tuna Thunnus orientalis during chilled storage

Using the full-cycle cultured (FC) Pacific bluefin tuna [body weight 16.3±1.9 kg (pre-fasting group, pre-FG), 14.2±0.9 kg (post-fasting group, post-FG)], changes in the physical/chemical properties of the cephalal parts of dorsal (D) and ventral (V) ordinary muscles (OM) by fasting (6 days) during chilled storage (4°C) were investigated. Condition factors were 26.7 (pre-FG) and 20.3 (post-FG, P < 0.05). Fasting changed the liver color to green. Fasting also decreased the amount of protein and lipid contents of the DOM and VOM of FC tuna. The breaking strength and pH of the DOM and VOM of post-FG tuna were higher (P <0.05) than for pre-FG tuna during storage. In contrast, the glycogen contents of DOM and VOM of post-FG tuna were lower than for pre-FG tuna. The color values (L*, a* and b*) of DOM of post-FG tuna were lower than for pre-FG tuna throughout the storage period. In addition, the metmyoglobin (metMb) content of DOM of post-FG tuna was lower (P <0.05) than that of pre-FG tuna, and the metMb content of VOM of post-FG tuna remained low after fasting. These results indicate that fasting suppresses deterioration (especially meat color) of FC tuna muscles during chilled storage.     

Possibility for decreasing of mercury content in bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> by fish culture

ABSTRACT:   The bluefin tuna tested were reared for 22 months from eggs before the beginning of the experiment, and sampling was performed every 3 months over the following year. The experimental results showed that the mercury concentration in the muscle ranged from 0.32 to 0.85 μg/g, which is lower than that found in wild bluefin tuna of a similar size. Increase in the mercury concentration corresponding to the increase in body weight was not shown, and it was quite different with wild bluefin tuna. Furthermore, no significant relationship was found between the lipid concentration and the mercury concentration in muscle. Among the internal organs of cultured bluefin tuna, the heart (0.32–0.66 μg/g), liver (0.43–0.99 μg/g) and spleen (0.59–1.0 μg/g) contained higher concentrations of mercury. It was estimated that the full-cycle cultured bluefin tuna had been fed small fish containing lower concentrations of mercury, and that the mercury concentration of tuna would be almost equal throughout the year because the effect of mercury accumulation would be weakened by body growth. Therefore, it was concluded that selecting diet fish species might decrease mercury contents in cultured bluefin tuna.     

Sizes of adult bluefin tuna Thunnus orientalis in different areas of the western Pacific Ocean

ABSTRACT:   The length frequencies of adult Pacific bluefin tuna Thunnus orientalis caught near Japan, near Taiwan, in the tropical western Pacific Ocean, and in the South Pacific Ocean, were compared. The largest catches consisted of medium-sized adults (160–209 cm fork length) taken near Japan and large adults (≥210 cm) taken near Taiwan. Small adults (120–159 cm) were occasionally caught in the Sea of Japan. Small numbers of medium-sized and large adults were caught in the tropical western Pacific and the South Pacific. The length frequencies of the fish greater than 160 cm in length caught in different areas around Japan were similar to each other. As adult bluefin tuna grow older they spawn earlier, and their distribution, movements, and spawning areas change.     

Comparison of the proximate compositions,breaking strength and histological structure by the muscle positions of the full-cycle cultured Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:   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.     

Distribution,growth and hatch date of juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> in the coastal area of the sea of Japan

ABSTRACT:   Mid-water trawl surveys were conducted from late August to late September in 1999 and 2004 in order to investigate the distribution pattern, hatch date, and growth of juvenile Pacific bluefin tuna Thunnus orientalis in the Sea of Japan. Juveniles were collected at the stations where ambient water temperature (mean water temperature from surface to 30 m deep, WT₀₋₃₀) was 23.4–25.9°C, and most of them were found in waters where WT₀₋₃₀ was higher than 24°C. Sampled juveniles ranged 108–280 mm fork length. Based on otolith analysis, they were estimated to grow to approximately 180 and 250 mm at 60 and 90 days old, respectively, and showed similar growth to that of Atlantic bluefin tuna in the Mediterranean Sea. The back-calculated hatch date of the samples was mostly in July and most juveniles spawned in the Sea of Japan.     

Turbulence effect on survival and feeding of pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> larvae,on the basis of a rearing experiment

ABSTRACT:   Physical conditions such as oceanic turbulence related to food availability are considered to be important factors affecting fish larval survival. Rearing experiments were conducted to elucidate the effects of turbulence on the survival and feeding rates during the initial feeding period of Pacific bluefin tuna Thunnus orientalis . Six levels of turbulence intensity were provided by changing flow rates from pipes set on the bottom of rearing tanks. The result showed a dome-shaped relationship between turbulence level and survival rate, in which the feeding rate appeared higher at a logged turbulence energy dissipation rate of −6.32, and decreased at both higher and lower turbulence levels. Compared with the turbulence intensity in the ocean, the optimal turbulence level for Pacific bluefin tuna larvae corresponded to the turbulence caused by sea surface winds with speeds of 4–12.5 m/s. The estimated optimal turbulence intensity for Pacific bluefin tuna larvae is comparable to that for yellowfin tuna Thunnus albacares .     

Ontogenetic changes in RNA,DNA and protein contents of laboratory-reared Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:     The ontogenetic changes in the growth potential of larval and juvenile laboratory-reared 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.     

Structural and thermodynamic characterization of tropomyosin from fast skeletal muscle of bluefin tuna

ABSTRACT:   Tuna tropomyosin is a mixture of nearly equimolar amounts of two isoforms (designated α and β). cDNA encoding the α form was cloned from bluefin tuna Thunnus thynnus fast skeletal muscle. The full-length cDNA contained 1220 bp, comprising an open reading frame of 855 bp encoding 284 amino acid residues, flanked by 5'-untranslational regions (156 bp) and 3'-untranslational regions (209 bp). The deduced amino acid sequence showed considerably high homology in a range of 93.7–98.6% to those of other vertebrate α-type tropomyosins. In phylogenetic analysis, bluefin tuna tropomyosin showed the closest relationship with the white croaker counterpart. The predicted mass was 32 919 Da, and isoelectric point was 4.50, assuming acetylation of the N-terminus. By differential scanning calorimetry, bluefin tuna tropomyosin gave two major endothermic peaks at 29.3 and 41.5°C, probably caused by the presence of two isoforms. Circular dichroism spectra supported such a unique denaturation profile.     

Muscle fiber types,growth and development in the whole myotome of cultured Pacific bluefin tuna Thunnus orientalis

Following the successful development of Pacific bluefin tuna (PBT) aquaculture, it is of considerable importance to determine the muscle fiber types and their growth patterns for future development. Muscle fiber profiles of dorsal ordinary, lateral ordinary (LO) and dark muscles and their growth patterns in PBT from 3.0 to 54.3 kg body weight were studied. Muscle fibers were histochemically stained for NADH-diaphorase and myosin adenosine triphosphatase activity (mATPase), and immunohistochemically stained with S-58 slow-muscle myosin antibody. All muscle fibers in dorsal and LO muscles showed low NADH-diaphorase activity, and acid-labile (pH 4.0 or 4.3) and alkali-stable mATPase activity. In LO muscle adjacent to dark muscle, three intensities of mATPase activity were observed after acid pre-incubation at pH 4.5 or 5.0, and the activity was related to the muscle fiber diameter. In dark muscle, all small and some large fibers stained intensely for NADH-diaphorase activity, related to their high aerobic metabolism. The high-active fibers with NADH-diaphorase in dark muscle were positive for S-58 antibody. Some large fibers in dark muscle showed intermediate NADH-diaphorase activity and high mATPase activity after alkali pre-incubations. These are fast-twitch oxido-glycolytic fibers in dark muscle and transformed to red muscle fibers with increasing body weight.     

Primary structure and thermostability of bigeye tuna myoglobin in relation to those of other scombridae fish

ABSTRACT:   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.     

In vitro amino acid absorption using hydrolysed sardine muscle or soybean meal at different intestinal regions of the Pacific bluefin tuna (Thunnus orientalis)

The amino acid (AA) absorption along the intestinal tract of the Pacific bluefin tuna (Thunnus orientalis) was evaluated using two hydrolysed protein sources (fresh sardine muscle and soybean meal) with the everted intestine technique. Pork pepsin and pancreatic enzyme extract from the bluefin tuna were used to hydrolyse the protein from fresh sardine (FSH) and soybean meal (SMH) under optimal bluefin tuna fish physiological conditions. Both of the hydrolysate solutions were tested within three intestinal sections from the bluefin tuna. The everted intestinal fractions immersed in the hydrolysate solutions were sampled at different times to analyse for AA and absorption rate calculations. Fresh sardine and SMH contained greater amounts of essential amino acids (EAA) than those of non‐essential amino acids (NEAA); however, the profiles of AA absorbed showed higher absorption of NEAA in both cases. Using a similar concentration solution, the absorption rates within the intestinal fractions showed a preferential absorption in the proximal and distal regions for Arg and His when FSH was used. However, the absorption rates for Lys resulted in a decreasing proximal‐to‐distal gradient between the different intestinal regions for FSH and SMH. The possibility of a catabolic role of certain AAs in the enterocytes being able to explain the differences in absorption is discussed.     

Recruitment abundance index of Pacific bluefin tuna using fisheries data on juveniles

ABSTRACT:   The recruitment abundance index of Pacific bluefin tuna Thunnus orientalis was estimated from 1980 to 2003 fishing year by using the troll fishery data in Nagasaki Prefecture, western Japan. It has been shown that the troll fishery in Nagasaki Prefecture operates with good time–area coverage of the species habitat, and that the fishing power slightly changed during the period analyzed, based on fisheries statistics, published information, and interviews with the fishers. Average catch per unit effort (CPUEs) were standardized by a generalized linear model (GLM) considering the effects of fishing year, season and landing area. Standardized CPUE of age-0 bluefin tuna showed larger fluctuations year by year than the nominal CPUE combined for all ages. High CPUEs in fishing years of 1981, 1994, 1996 and 1999 were observed. Data from these years agreed with the higher recruitments estimated by virtual population analysis (VPA) or higher catch of age-0 fish reported for the Pacific side. The age-specific standardized CPUE of age-0 bluefin tuna in this study was judged to be a useful indicator of recruitment.     

Changes in the scotopic vision of juvenile Pacific bluefin tuna (Thunnus orientalis) with growth

In cultured juvenile Pacific bluefin tuna (Thunnus orientalis), reducing the mass deaths caused by collision or contact with tank or net walls at night is a priority for seedling production. Pacific bluefin tuna is a visually dependant species, although its scotopic vision is poor. We recorded electroretinograms to investigate the visual function with growth in the dark-adapted eyes of juvenile Pacific bluefin tuna. Peak wavelengths of spectral sensitivity [38–62 days posthatch (dph), 77–167 mm standard length (SL)] were observed between 474 and 494 nm. Visual light sensitivity has a tendency to increase slightly with growth at 28–64 dph in individuals that measured 29–175 mm SL. However, visual temporal resolution did not significantly increase with growth at 38–62 days dph in individuals that measured 77–167 mm SL. These results suggest that the mass death continues between 28 and 64 dph because of low visual function and increasing swimming speed with growth.     

Decadal variation in the trans-Pacific migration of northern bluefin tuna (Thunnus thynnus) coherent with climate-induced change in prey abundance

Northern bluefin tuna, Thunnus thynnus, apparently spawn only in the western Pacific and a portion of the juveniles migrate to the eastern Pacific. During the past decade, catches of northern bluefin in the eastern Pacific have declined. One possible cause for this decline, proposed by bluefin stock assessment studies, is a decline in the proportion of bluefin that migrate out of the western Pacific. This hypothesis is examined with several indices of the relative abundance of bluefin tuna in the western and eastern Pacific. These indices suggest a decline in the proportion of bluefin migrating to the eastern Pacific since 1977. This period of reduced bluefin migration coincides with a period when a prey of bluefin, Japanese sardine, Sardinops melanosticta, were abundant off Japan. It is hypothesized that in years when sardines are abundant off Japan, a higher proportion of bluefin stay in the western Pacific compared with years when sardines are scarce. Currently, the adun-dance of sardines off Japan is declining. If this decline continues, this hypothesis predicts an increase in bluefin migrating north of Hawaii and into the eastern Pacific.     

Muscle growth in full-cycle cultured Pacific bluefin tuna Thunnus orientalis from early larval to juvenile stage: histochemical,immunohistochemical and ultrastructural observation

With the development of successful aquaculture techniques, the Pacific bluefin tuna (PBT) has become the most important fish species in Japan. Early muscle growth and cellularity of muscle fibers have a profound effect on ultimate body size and meat quality of fish. Larvae and juveniles of full-cycle cultured PBT were sampled at 3, 15, 29, 41, 70, 128 and 218 days post hatch (dph). Transverse body sections until 41 dph and muscle sections from 41 dph thereafter from dorso-cranial or caudal areas were stained for muscle fiber morphological and morphometric analysis and muscle sections from the dorso-cranial region were used for ultra-structural observation. Red muscle fibers appeared by 15 dph at the horizontal septum both in epaxial and hypaxial regions. Other than a single layer of superficial red fibers and those at the horizontal septum, existence of red muscle fibers was apparent in the PBT myotome. The muscle fiber diameters varied in their size even in adjacent myotomes of PBT. Overall, growth throughout the various stages was a result of both hyperplasia and hypertrophy of muscle fibers. Muscle fiber diameters were greater in the dorso-cranial than in dorso-caudal region at 41, 70 and 128 dph. These results provide additional characterization of PBT growth, which may assist with managing fish production efficiency, harvest age and eating quality.     

Thermal adaptation of pacific bluefin tuna Thunnus orientalis to temperate waters

ABSTRACT:   Immature Pacific bluefin tuna Thunnus orientalis , tagged with archival tags, were released near Tsushima Island in the East China Sea during the winters of 1995 through 1998. Time-series data for ambient and peritoneal cavity temperatures, recorded every 128 or 256 s for 23 fish recovered, were analyzed. The objective of this study was to clarify the process of development of thermoconservation ability with growth in relation to adaptive mechanisms to cooler temperate waters. According to the results, mean ambient temperatures ranged from 14.9 to 20.7°C, which is almost within the optimum temperature range according to previous reports. Mean peritoneal temperatures were higher than ambient temperatures (19.7–27.3°C), but never reached 35°C, which would induce overheating. Although the mean thermal differences between peritoneal and ambient temperatures increased with body size, the rate of increase decreased with body size. A heat budget model suggests that as the insulation of the body develops, the estimated mean values of internal heat production decrease with body size. This is probably due to the allometric scale effect and explains why the thermal difference does not increase quickly with body size. It is likely that Pacific bluefin tuna inhabit cooler temperate waters in mid-latitude regions to avoid overheating.     

Genotyping‐by‐sequencing for construction of a new genetic linkage map and QTL analysis of growth‐related traits in Pacific bluefin tuna

Pacific bluefin tuna (Thunnus orientalis) has high market value, but its wild populations have decreased in recent years. The broodstock of Pacific bluefin tuna that were hatched artificially and reared under aquaculture conditions is beginning to be used for production. The creation of broodstock with commercially valuable traits, such as rapid growth, is therefore of great interest. Genetic linkage map‐based identification of markers associated with quantitative trait loci (QTLs) facilitates marker‐assisted selection (MAS) breeding and allows efficient genetic improvement of broodstock. Single nucleotide polymorphism (SNP)‐based genetic linkage map construction using the genotyping‐by‐sequencing method can expand the number of mapped markers and help identify growth‐related QTLs. In this study, we constructed sex‐specific maps for 24 linkage groups consisting of 677 SNP and 651 microsatellite markers. The total lengths of 93 progenies in the mapping population followed normal distribution, with an average length of 9.4 mm. We performed composite interval mapping in the mapping population. QTL analysis revealed one significant QTL in LG10 on the female linkage map. The genetic linkage map—the second such map generated for Pacific bluefin tuna—and the growth‐related QTLs detected in this study will be useful for tuna aquaculture MAS programs.     

Purification and characterization of glutathione peroxidase 1 in the red muscle of Pacific bluefin tuna Thunnus orientalis

Glutathione peroxidase GPx1 was purified from the red muscle of Pacific bluefin tuna Thunnus orientalis and its enzymatic properties were characterized. The enzyme was optimally active at pH 7.4 with a specific activity for hydrogen peroxide and a K _m of 6.7 μM. cDNA was also isolated and it contained a predicted open reading frame (ORF) encoding a 188 amino acid protein. The phylogenetic tree shows that fish including Pacific bluefin tuna, pufferfish, and zebrafish, but not mammals, possess two genetically distinct types of GPx1, i.e., GPx1a and GPx1b. The purified enzyme was classified as a fish GPx-1b enzyme on the basis of the phylogenetic tree of the GPx1 family.