Dietary utility of enzyme-treated fish meal for juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:   In order to develop an artificial diet, the dietary utility of enzyme-treated fish meal was investigated for juvenile Pacific bluefin tuna Thunnus orientalis (PBT). Diets containing each 63% of Chilean fish meal (FM), enzyme-treated Chilean fish meal (EC) and enzyme-treated Peruvian fish meal (EP), with 10% bonito oil and raw sand lance Ammodytes personatus (SL) were fed to juvenile tuna six times per day for one week. In a different trial, diets EC and SL were fed to tuna six times per day for 2 weeks. Only diet EC sustained similar growth or caused lower survival and higher feed efficiency, hepato- and enterosomatic indices and final carcass lipid content as compared to those of SL. Diets FM and EP led to lower specific growth rate (SGR) but similar feed efficiency, survival and hepatosomatic index, yet higher enterosomatic index. Moreover, PBT fed diet EC for 2 weeks led to similar growth performance but higher final carcass and hepatic lipid contents, and plasma cholesterol and phospholipid levels than those fed SL. Carcass fatty acid composition of diet EC group had lower 20:5  n -3 and 22:6  n -3 levels than the SL group. These results revealed that EC, as a suitable dietary protein source, could sustain growth of PBT, while dietary bonito oil led to higher carcass lipid but lower accumulation of n -3 highly unsaturated fatty acids.     

Partial characterization of the digestive enzymes of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> under culture conditions

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.     

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.     

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.     

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.     

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.     

Whole-body heat transfer coefficient and body temperature change of juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> 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.     

Three cone opsin genes and cone cell arrangement in retina of juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:   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).     

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 .     

Development of visual cells in the Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

The development of rod and cone photoreceptor cells was investigated in the retinas of Pacific bluefin tuna larvae and juveniles, using RET-P1 monoclonal antibody labeling to identify photoreceptors. At 60 h after hatching, which was about when feeding began, opsin (presumably green opsin (Rh2)) was expressed in the outer segments of cone cells. At 15 days after hatching (dah), although many labeled cone cells were observed in the dorsal retina, the same type of cone cells had partially appeared in the ventral retina. The presence of rod cell bodies was confirmed by the expression of Rh1 opsin at 15 dah. At 21 dah, the presence of outer segments of rod cells was confirmed by the expression of Rh1 opsin and by morphology. The observations suggest that the cone cells were substantially operable upon the development of their outer segment at around the beginning of the post-larval stage, and the rod cells began to function at around 15 to 21 dah, before and during metamorphosis.     

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.     

The changes in proximate compositions and glycogen contents in the dorsal ordinary muscles of the full-cycle cultured Pacific bluefin tuna Thunnus orientalis occurring with growth

ABSTRACT:   Using full-cycle cultured (FC) Pacific bluefin tuna (body length [BL], 42.6–66.4 cm; body weight [BW], 1.66–7.40 kg, n  = 15), the changes in chemical compositions and histological structure of the cephalal parts of the dorsal ordinary muscles (DOM) occurring with growth were investigated. A positive correlation ( r  = 0.9644, P  < 0.05) was observed between BL and BW with growth. The protein, lipid and ash contents of DOM and condition factor did not change with growth. However, the glycogen content of DOM increased ( P  < 0.05) from approximately 55 cm (BL) in this study. Using optical microscopic photographs, the various shapes of muscle fibers were observed and it was noted that the muscle fiber diameter increased ( P  < 0.05) with growth. Using transmission electron microscopic observation, many glycogen granules were observed in muscle fibers (especially, side of connective tissue) of DOM throughout the growth stage in this study. These results indicate that the glycogen content of DOM of FC Pacific bluefin tuna increases before the lipid with growth.     

Myxosporean and microsporidian infections in cultured Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> in Japan

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.     

Effect of water temperature on the feeding activity and the resultant mercury levels in the muscle of cultured bluefin tuna Thunnus orientalis (Temminck and Schlegel)

Tuna muscle often contains high levels of mercury, and fish samples with mercury concentrations ten times higher than the specified safety standards have been reported. Here, we report on the relationship between water temperature and the concentration of mercury in the tail muscle tissue of cultured bluefin tuna Thunnus orientalis. The fish used in this study were cultured at Fisheries Laboratory of Kinki University (Amami Experimental Station, Kagoshima, Japan). One hundred fish weighing 26.2–89.4 kg were selected for analysis between February 2007 and January 2008. Water temperature during rearing ranged from 21 to 29 °C. The total mercury levels were measured using the reduction vaporizing atomic absorption method after acid digestion. Body weight increased approximately 1.5 times that observed in a previous study, despite feeding activity either being the same or less than that observed previously. The average mercury concentration in white muscle was 0.353 mg kg^?1, remaining almost constant and independent of body growth. Unlike previous studies, seasonality was not observed in this study. Based on these findings, water temperatures within a certain range were considered to stabilize feeding activity and increase feeding efficiency. Consequently, water temperature is considered to have a moderating effect on seasonal fluctuations in muscle mercury concentrations in cultured bluefin tuna.