Energy partitioning in cultured juvenile Pacific bluefin tuna,Thunnus orientalis (Temminck & Schlegel, 1844)

The characteristics of dietary utilization, energy conversion efficiency, metabolic rate and energy partitioning were measured for cultured Pacific bluefin tuna (PBT) juveniles fed on an artificial diet. Thirty‐one juveniles (1.1 ± 0.3 g BW) were stocked into each of two 2500 L tanks to measure oxygen consumption (?O₂), swimming speed, digestibility and growth performance. ?O₂ elevated until 2.5 ± 0.3 times of pre‐feeding level within 1.5 h after feeding, except for the first feeding, and returned to pre‐feeding level within 3 h. Swimming speed fluctuation was corresponded with the ?O₂ fluctuation, and both parameters were stable from 02:00 to 06:00 and also during the whole day for starved fish. These indicate that feeding has strong influence on their metabolic rate. Energy partitioning for faecal, urinary and branchial, heat increment and voluntary activity, standard metabolism, and retained energy were calculated to be 17.2%, 5.9%, 14.9%, 41.3% and 20.7% of total ingested energy, respectively. The results indicate that, unlike other fish, juvenile PBT distribute large amount of energy for maintenance, which allows only a little proportion of ingested energy available for growth.     

Gustatory responses in Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel)

Gustatory neural responses of the Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel) to extracted compounds of prey organisms, such as amino acids, nucleotide‐related substances, organic acids and organic bases, were electrophysiologically recorded from the facial nerve supplying the anterior palate. Of the 17 amino acids tested, l ‐proline was the most potent amino acid at 10^?2 M, and its threshold was the lowest at around 10^?6 M. l ‐leucine, l ‐methionine, l ‐alanine, l ‐valine and l ‐isoleucine were also highly stimulatory at 10^?2 M; however, the other 11 amino acids examined were not as effective or were ineffective. Thus, the gustatory receptors for amino acids of the Pacific bluefin tuna show a narrowly tuned response profile. Among the seven nucleotide‐related substances tested, uridine‐5′‐monophosphate, inosine‐5′‐monophosphate and adenosine‐5′‐diphosphate were highly stimulatory, and their thresholds were 10^?4–10^?5 M. Inosine elicited a positive response at 10^?2 M but its response magnitude was not so high. Organic acids l ‐lactic and pyruvic acids were effective at 10^?2 M, but no response was elicited at 10^?3 M. Among organic bases, betaine was highly stimulatory, and its response magnitude at 10^?2 M is almost equal to that of l ‐proline at the same concentration. The threshold for betaine was determined to be at around 10^?5 M. Trimethylamine oxide and ammonium chloride were ineffective.     

Completion of the Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) life cycle

Tuna aquaculture is currently dependent on the wild capture of juveniles for production. The development of hatchery technology for bluefin and other tunas would be a major step forward in improving sustainability of their aquaculture. The present study overviews the technology in the life cycle completion of the Pacific bluefin tuna (PBT) Thunnus orientalis (Temminck et Schlegel) under aquaculture conditions in Kinki University, and the problems to be solved for the establishment of tuna hatchery technology. On 23 June 2002, broodstock of PBT that were artificially hatched and reared spontaneously spawned in captivity. The resulting eggs hatched and were subsequently reared to the juvenile stage. The spawning fish were the result of a research project started in 1987 to rear wild‐caught juvenile PBT that were several months old. Fertilized eggs were obtained from these fish in 1995 and 1996. Resulting juveniles (the artificially hatched first generation) were reared to maturity and spawned in 2002. Over the summer of 2002, 1.63 million eggs from these fish were used for a mass rearing experiment, and 17 307 juveniles were produced and transferred to an open sea net cage. Of these artificially hatched second‐generation PBT, 1100 grew to approximately 95 cm total length and 14 kg body weight in 22 months. This procedure means the completion of PBT life cycle under aquaculture conditions, which was first attained among large tuna species. The problems awaiting solution in PBT hatchery production are their unpredictable spawning in captivity, to improve survival during the first 10 days post hatch, to reduce cannibalism in larval and juvenile stages, and to solve collision problem causing high mortality during the juvenile stage.     

Ontogeny of immune system organs in northern bluefin tuna (Thunnus orientalis, Temminck and Schlegel 1844)

Serial sections, prepared from 0.5 to 30 days posthatch (dph) larval and juvenile Thunnus orientalis (Temminck & Schlegel 1844), were stained with haematoxylin and eosin and examined by light microscopy for immune organ development. The early kidney was present at 0.5 dph, undifferentiated stem cells began to appear at 2 dph, and by 7 dph occasional small lymphocytes were present. The thymus was first obvious at 5 dph, located above the fourth branchial arch, small lymphocytes appeared at 7 dph, and by 15 dph an outer thymocytic zone and an inner epithelioid zone were visible. The progenitor spleen was present at 2 dph, located close to the gut, and by 12 dph it consisted of a mass of sinusoids filled with red blood cells, and remained mainly erythroid throughout the period studied. These results suggest that development of immune organs in this species is precocious relative to other marine teleosts.     

Retinal ganglion cell topography in juvenile Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel)

The retinal ganglion cell distribution, which is known to reflect fish feeding behavior, was investigated in juvenile Pacific bluefin tuna Thunnus orientalis. During the course of examination, regularly arrayed cells with a distinctive larger soma, which may be regarded as motion-sensitive cells, were found. The topographical distribution of ordinary-sized ganglion cells, which is usually utilized to estimate fish visual axis and/or visual field characteristics, showed that the highest-density area, termed the area centralis, was localized in the ventral-temporal retina. The retinal topography of ordinary-sized ganglion cells seems to reflect the bluefin tuna’s foraging behavior; while cruising, cells in the area centralis may signal potential prey, such as small schooling pelagic fishes or squids, that are present in the upward-forward direction. Judging from morphological characteristics, the large ganglion cells localized in the small temporal retinal area seem to be equivalent to physiologically categorized off-center Y-cells of cat, which are stimulated by a transient dark spot in a bright visual field. It was inferred that presumed large off-center cells in the temporal retina detect movements of agile prey animals escaping from bluefin tuna as a silhouette against environmental light.     

Effect of Artemia enrichment on the growth and survival of Pacific bluefin tuna Thunnus orientalis (Temminck & Schlegel) larvae

This study was carried out to investigate the suitability of Artemia enriched with docosahexaenoic acid (DHA) and choline as live food on the growth and survival rate of the Pacific bluefin tuna (PBT; Thunnus orientalis) larvae. The PBT larvae were fed either Artemia enriched with oleic acid (Diet 1), DHA (Diet 2), DHA+choline 1.0 mg L^?1 (Diet 3) and DHA+choline 2.0 mg L^?1 (Diet 4) or striped knifejaw larvae (Diet 5, reference diet), in duplicate for 12 days. Enrichment of Artemia with DHA significantly increased the DHA levels to 13.9, 13.8 and 12.5 mg g^?1 on a dry matter basis in Diets 2, 3 and 4 respectively; however, the levels were significantly lower than the reference diet (26.9 mg g^?1 dry matter basis; Diet 5). Although growth and survival rate were significantly improved by the enrichment of Artemia with DHA and choline, the improvement was negligible compared with the enhanced growth and survival rate of the fish larvae‐fed group (P<0.05). The results demonstrated that enriched Artemia does not seem to be the right choice to feed the PBT larvae perhaps because of the difficulties in achieving the correct balance of fatty acid with higher DHA/EPA from Artemia nauplii.     

Promotion of initial swimbladder inflation in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

Initial swimbladder inflation (ISI) of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae was studied to increase the survival of cultured larvae. Experiment 1 was conducted to explore promotion and inhibition of ISI under different water surface conditions; including the use of surface skimmer to remove autogenous surface substances (SS), covering the water surface with liquid‐paraffin‐layer (LP) and oil film (OF), and a control (non‐treatment, NT). Significantly higher inflation frequency was observed in SS (62.2%) than NT (11.9%), LP (2.7%) and OF (3.9%). This indicates that ISI in PBT larvae can be promoted by removal of surface substances on rearing water which inhibit larval air gulping. Experiment 2 aimed to elucidate proper day of larval age to start skimming for promoting ISI with four different periods of oil film removal: from 3 to 8 (SF3D), 4 to 8 (SF4D), 5 to 8 (SF5D), 6 to 8 (SF6D) days‐post‐hatch (dph). Significant improvement in ISI frequency was observed in SF3D (80.2%) but the frequency was very poor in SF4D, SF5D, and SF6D (17.8–7.5%). This implies the need of oil film removal without missing a narrow window, 1 day of 3 dph, to promote ISI in practical PBT larviculture.     

Diel and ontogenetic body density change in Pacific bluefin tuna, Thunnus orientalis (Temminck and Schlegel), larvae

Diel and ontogenetic changes in larval body density related to swim bladder volume were investigated in Pacific bluefin tuna, Thunnus orientalis, to determine the causality of larval mortality – adhesion to the water surface and contact with the tank bottom during seedling production. The density of larvae with deflated swim bladders increased with total length and days post hatch. Diel density change was observed after day 2 post hatch; owing to daytime deflation and night‐time inflation of the swim bladder, the density was relatively higher during the daytime. Increased swim bladder volumes clearly reduced larval density during the night‐time after day 9 post hatch. However, the density of larvae with inflated swim bladders was greater than rearing water density (Δρ>0.0099). The small density difference between larvae and rearing water (Δρ=0.0022?0.0100) until day 4 post hatch may have caused larval mortality by adhesion to the water surface because larvae can be easily transported to the water surface by aeration‐driven upwelling in rearing tanks. Density increased noticeably from day 5 to day 9 post hatch. The increased density difference (Δρ=0.0065?0.0209) in larvae and rearing water possibly induced mortality by contact with the tank bottom because larvae sink particularly during the night‐time on ceasing swimming.     

Influence of initial swimbladder inflation failure on survival of Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

This study investigated the influence of initial swimbladder inflation (ISI) failure on survival of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae to prevent mass mortality due to sinking death. In Experiment 1, swimbladder inflation frequency and survival within an ISI promoted (PS) group, for which surface film of rearing water was removed, and a group without ISI promotion (NPS) were compared in 20 and 30 kL tanks. The PS group demonstrated significantly higher swimbladder inflation frequency and increased survival than the NPS group within 20 kL tanks at 9 days post hatching (dph). Similar tendencies were observed within 30 kL tanks. In Experiment 2, larval distribution and swimbladder inflation frequency in the night‐time were examined through larval sampling within the upper and middle layers of the water column and tank bottom within 1.6 and 30 kL tanks at 5 dph. Larvae at tank bottom had higher distributional density and significantly lower swimbladder inflation frequency than those distributed in the upper and middle layers within 1.6 kL tanks. Similar tendencies were observed within 30 kL tanks. Results of this study suggest the improvement of larval survival due to prevention of sinking death through ISI promotion in mass‐scale PBT larviculture.     

Mortality processes of hatchery‐reared Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) larvae in relation to their piscivory

In mass culture of Pacific bluefin tuna Thunnus orientalis, yolk‐sac larvae of other species are fed as a major prey item to tuna larvae from 7 to 8 mm in total length. Marked growth variations in tuna larvae are frequently observed after feeding of yolk‐sac larvae, and this variation in the growth of tuna larvae is subsequently a factor leading to the prevalence of cannibalistic attacks. To elucidate details of the mortality process of hatchery‐reared tuna larvae after the initiation of yolk‐sac larvae feeding, we compared the nutritional and growth histories of the surviving (live) tuna larvae to those of the dead fish, found dead on the bottom of the tank, as direct evidence of their mortality processes. Cause of mortality of tuna larvae 3 and 5 days after the initiation of feeding of yolk‐sac larvae was assessed from nitrogen stable isotope and otolith microstructure analyses. Stable isotope analysis revealed that the live fish rapidly utilized prey fish larvae, but the dead fish had depended more on rotifers relative to the live fish 3 and 5 days after the initiation of feeding of yolk‐sac larvae. The growth histories based on otolith increments were compared between the live and dead tuna larvae and indicated that the live fish showed significantly faster growth histories than dead fish. Our results suggest that fast‐growing larvae at the onset of piscivory could survive in the mass culture tank of Pacific bluefin tuna and were characterized by growth‐selective mortality.     

Optimal periods of night‐time lighting in the sea cage culture of Pacific bluefin tuna,Thunnus orientalis,juvenile (Temminck and Schlegel)

We have previously reported that night‐time lighting prevents the mass death of Pacific bluefin tuna (PBT) juvenile due to collision and/or contact with the walls of sea cages, immediately after transfer to the cages, and that night‐time lighting does not negatively impact fish stress levels. However, the period of night‐time lighting should be limited to minimize negative impacts on the surrounding environment and aid management. Therefore, we investigated the optimal period of night‐time lighting by evaluating the whole‐body cortisol and glucose levels as stress parameters, growth performance and survival of PBT juvenile in four cages with different periods of night‐time lighting (i.e. unlit, 4‐day, 8‐day and 12‐day lighting). The results showed that almost all fish were died 1 day after transfer to the unlit cage. In comparison, the other groups (4‐day, 8‐day and 12‐day lighting) had high survival rates (92.5–96.0%) without significant difference. However, in the 4‐day‐lighting group, an obvious stress response was recorded on day 5, and growth performance was significantly lower. In the 8‐day‐lighting group, whole‐body cortisol levels were slightly elevated on day 9; however, significantly elevation was not recorded on day 12. These results indicate that the recommended lighting period of night‐time lighting in sea cages is 8–12 days.     

Effect of taurine enrichment in rotifer (Brachionus sp.) on growth of larvae of Pacific bluefin tuna Thunnus orientalis (Temminck & Schlegel) and yellowfin tuna T. albacares (Temminck & Schlegel)

To investigate the dietary effect of taurine on the larval stage of tuna species, Pacific bluefin tuna (PBF) and yellowfin tuna (YFT), larvae were reared until 16 days after hatching (dAH) and 14 dAH, respectively, and replicate samples were fed either non‐taurine‐enriched rotifers (T‐0) or rotifers enriched with 800 mg taurine L^?1 (T‐800). Most PBF and YFT larvae were at the preflexion stage until 7 and 8 dAH, and there were no differences in the growth performance and total protein content of larvae between the T‐0 and T‐800 groups (t‐test; P > 0.05). Thereafter, however, for larvae of both species, these parameters in the T‐800 group significantly increased with enhanced notochord development compared to those in T‐0 group (t‐test; P < 0.05). Except for the RNA content in PBF larvae, there were no significant differences in changes of DNA and RNA content with larval growth between the T‐0 and T‐800 groups, but both PBF and YFT larvae showed increased protein DNA^?1 and protein RNA^?1 ratios in the T‐800 group compared to the T‐0 group after notochord flexion. This indicates that taurine is an important nutrient for the rapid growth of early stage PBF and YFT larvae, and we conclude that the growth improvement of PBF and YFT larvae by dietary taurine supplementation is due to the increase in protein synthesis efficiency after notochord flexion.     

Improving rearing performance in sea‐cage culture of Pacific bluefin tuna Thunnus orientalis juveniles (Temminck and Schlegel) using small sea cages

High levels of mortality occur in large net‐cage culture of Pacific bluefin tuna (PBF) Thunnus orientalis due to poor growth on commencement of sea‐cage culture obstructing the mass production of fingerlings. Therefore, we carried out this study to develop a countermeasure by using small sea cages (square with 13‐m sides). PBF juveniles were directly transferred to a 30‐m‐diameter cage (control) and compared them to fish transferred to small sea cages for 12 days before being merged into another 30‐m‐diameter cage. The survival rate, growth performance and potential factors increasing mortality were examined. The results of our study showed that survival rate in small sea cages was approximately 20% higher than that of the control. Poor growth also occurred in the small sea cages; however, its frequency and daily mortality rate were half those in the control. In addition, growth performance such as specific growth rate and weight gain of PBF juveniles in small sea cages significantly increased compared to the control. These results suggest that using small sea cages could be an effective countermeasure for poor growth, which may mitigate the high‐mortality conditions of current sea‐cage culture systems for PBF.     

Timing to promote initial swimbladder inflation by surface film removal in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

This study investigated the optimal timing of day to promote initial swimbladder inflation (ISI) for improved Pacific bluefin tuna (PBT), Thunnus orientalis, larval survival. Larval swimbladder inflation frequency was compared based on three experiments using different time schemes of surface film removal (SFR) from 3 to 9 days post hatch (dph). SFR was conducted from 05:00 to 19:00 hours (light period: S.5–19), 19:00 to 05:00 hours (dark period: S.19–5), 08:00 to 19:00 hours (S.8–19) and the entire day (S.24) in Experiment 1; from 08:00 to 19:00 hours (S.8–19‐E2), 08:00 to 13:00 hours (S.8–13), 13:00 to 19:00 hours (S.13–19) in Experiment 2; and from 13:00 to 16:00 hours (S.13–16), 16:00 to 19:00 hours (S.16–19), 18:00–19:00 hours (S.18–19) in Experiment 3. The swimbladder inflation frequency at the experiment termination (9 dph) was significantly higher (P < 0.001) in S.24 (91.1 ± 5.7%), S.5–19 (92.2 ± 5.1%) and S.8–19 (93.3 ± 3.4%) than in S.19–5 (11.1 ± 5.1%) in Experiment 1, and remarkably higher in S.8–19‐E2 (81.7%) and S.13–19 (88.3%) than in S.8–13 (0.0%) in Experiment 2, and significantly higher (P < 0.001) in S.16–19 (84.4 ± 5.1%) and S.18–19 (70.0 ± 12.0%) than in S.13–16 (7.8 ± 3.9%) in Experiment 3. These results suggest that the optimal timing to promote larval ISI by SFR is a few hours before the end of light period (16:00–19:00 hours) from 3 to 9 dph.     

Effects of feeding conditions and size differences on aggressive behaviour and cannibalism in the Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel) larvae

We examined the effects of feeding conditions and size differences on the aggressive behaviour and cannibalism in Pacific bluefin tuna (PBT) Thunnus orientalis larvae. In a 24 h experiment, restricted feeding alone was found to remarkably increase the frequency of aggressive behaviour, which was further elevated by differences in fish size. In a 4‐day rearing experiment, while aggressive behaviour was increased by restricted feeding alone, the frequency of cannibalism did not change significantly. Although the frequency of aggressive behaviour did not increase with difference in size factor, small fish in this group gradually tended to die over 4‐day period. In the restricted feeding and size difference group, large fish were observed to bite the small fish, and almost all the small fish died on the day after the start of the experiment. These results suggest that the aggressive behaviour of PBT larvae is chiefly increased by the shortage of live food; however, deaths related to cannibalism mainly occurred in small larvae and rapidly increased with food restriction and differences in fish size.     

Effects of night‐time light intensity on the survival rate and stress responses in juvenile Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel)

Land‐based cultured juvenile Pacific bluefin tuna Thunnus orientalis (PBT) have high mortality rates due to collisions or contacts with tank walls after about 30 days of hatching. To determine the effect of night‐time lighting on their survival, juvenile PBT were reared under four different night‐time light intensities (0, 5, 15 and 150 lx) for 9 days, followed by a 3‐day observation period. High‐intensity, night‐time lighting (150 lx) significantly improved the survival rate (75.8%; P < 0.001) compared with the unlit control group (0 lx, 64.3%). The survival rate in the high‐intensity group decreased after the end of the lighting period. Lighting did not influence whole‐body cortisol levels, glucose levels, or diel changes in plasma cortisol levels. In contrast, the survival rates of fish exposed to light intensities between 5 and 15 lx were slightly lower than that of the unlit control group. These results suggest that providing night‐time lighting of 150 lx or higher is an effective method for reducing the mortality of cultured PBT.     

Effect of tank wall colour and pattern on the survival rate of juvenile Pacific bluefin tuna Thunnus orientalis (Temminck and Schlegel) during ship transportation

Juvenile Pacific bluefin tuna (Thunnus orientalis; PBT) often experience high mortality during ship transportation. This study investigated whether the addition of colours or patterns to the walls of tanks affected survival rate. In the first experiment, three colours and lattice patterns were tested: dark blue single‐colour, red single‐colour, and red–blue lattice pattern. Fish in all tanks exhibited abnormal behaviours when sunlight entered the tanks between 0800 and 1000 hours, but mortality only increased in the single‐coloured tanks as a result of collision with the tank walls. In the second experiment, four colours and patterns were tested: dark blue single‐colour, red–blue lattice pattern, red–blue lattice pattern with shade sheet and red–green lattice pattern with shade sheet. Again, we visually observed that fish in all treatment groups exhibited abnormal behaviour when sunlight entered the tanks, but there were no collision deaths in the lattice‐patterned tanks and survival in this group was significantly higher than in the single‐coloured tanks. Thus, the use of a high‐contrast colour pattern can prevent mass death of juvenile PBT during ship transportation.     

Visual spectral sensitivity of photopic juvenile Pacific bluefin tuna (Thunnus orientalis)

Although Pacific bluefin tuna is a species that relies on vision, its photopic visual function is not well known; we therefore recorded electroretinograms to investigate photopic spectral sensitivity in juveniles of this species (49-81?days post-hatch; standard length 74-223?mm). The peak spectral sensitivity wavelength was 505?nm. We estimated that two (λ(max)?=?512-515?nm and 423-436?nm) or three (λ(max)?=?512-515?nm, 423-436?nm, and 473?nm) types of cone visual pigments contribute to photopic vision; these spectral sensitivities are adapted to surface water habitats in clear ocean and coastal water.     

Influence of swimbladder inflation failure on mortality,growth and lordotic deformity in Pacific bluefin tuna,Thunnus orientalis, (Temminck & Schlegel) postflexion larvae and juveniles

This study examined the influence of swimbladder inflation (SBI) failure on mortality (Experiment 1), lordotic deformity (Experiment 2) and growth (Experiment 1, 2) in Pacific bluefin tuna, Thunnus orientalis, postflexion larvae and juveniles by producing larvae lacking inflated swimbladders. Experiment 1 was conducted for postflexion larvae to juveniles (from 18 to 30 days‐post‐hatch; dph). Mortality was not significantly different between the fish with (WIS) and without (WOIS) inflated swimbladders. Standard length (SL) and body weight (BW) were significantly smaller in WOIS than WIS. Moreover, the SBI was found in WOIS after postflexion stage. In Experiment 2, two examination trials were conducted on SBI, vertebral deformity and growth for juvenile stage. Lordotic deformities were found neither in the WOIS nor in the WIS. Although SL and BW were significantly smaller in WOIS than WIS at 22 dph, after 37 dph no significant differences were found between them. The results of Experiment 1 and 2 indicate that SBI failure causes growth retardation until juveniles of 30 dph; however, it does not cause growth retardation in juveniles after 37 dph and mortality of postflexion larvae and juveniles. Moreover, SBI failure did not cause lordotic deformity in PBT differently from many other fish.