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.     

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.     

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.     

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.     

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.     

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.     

Effects of photoperiod and night‐time aeration rate on swim bladder inflation and survival in Pacific bluefin tuna,Thunnus orientalis (Temminck & Schlegel), larvae

Success of swim bladder inflation (SBI) is crucial for early survival of Pacific bluefin tuna (PBF) larvae, because it reduces larval sinking death by enhancing buoyancy. In Experiment 1, we examined the effect of photoperiod on SBI and survival in PBF larvae by comparing photoperiods of 9L: 15D (9L), 14L: 10D (14L: natural photoperiod), 19L: 5D (19L) and 24L: 0D (24L) during 2–10 days post hatch (dph). In Experiment 2, the combined effects of photoperiod (24L and 14L) and nighttime aeration rate (enhanced night‐time aeration: ENA of 1300 mL min^?1 as a countermeasure for sinking death and 130 mL min^?1) on the survival and SBI were also examined during 2–10 dph. Moreover, in Experiment 3 the effect of photoperiod on vertical distribution of larvae in night‐time was examined on 3–5 dph. Photoperiod of 24L in Experiment 1 significantly inhibited SBI compared with 14L and 19L; nevertheless, it significantly improved survival compared with other photoperiods with a dark period. On the other hand, the shortened light period (9L) showed significantly reduced SBI and also survival. In Experiment 2, the countermeasure for sinking death of ENA under 24L did not further improve the survival; rather it tended to reduce the survival. In Experiment 3, larvae distributed less in the bottom layer in 24L than in 14L, suggesting the reducing effect of 24L on sinking death. The results indicate that 24L without ENA is suitable for survival which is the most serious problem in PBF larviculture.     

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.     

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

ABSTRACT:   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.     

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.     

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.     

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.     

Relationship between prey utilization and growth variation in hatchery‐reared Pacific bluefin tuna,Thunnus orientalis (Temminck et Schlegel), larvae estimated using nitrogen stable isotope analysis

In mass culture of Pacific bluefin tuna Thunnus orientalis, a marked growth variation is observed after they start feeding at 6–7 mm in body length (BL) on yolk‐sac larvae of other species, and the growth variation in tuna larvae is a factor leading to the prevalence of cannibalism. To examine the relationship between prey utilization and growth variation, nitrogen stable isotope ratios (δ¹⁵N) of individual larvae were analysed. A prey switch experiment was conducted under two different feeding regimes: a group fed rotifers (rotifer fed group), and a group fed yolk‐sac larvae of spangled emperor, Lethrinus nebulosus (fish fed group) from 15 days after hatching (6.87 mm BL). The fish fed group showed significantly higher growth than the rotifer fed group. Changes in the δ¹⁵N of the fish fed group were expressed as an exponential model and showed different patterns from those of the rotifer fed group. The δ¹⁵N of fast‐growing tuna larvae collected in an actual mass culture tank after the feeding of yolk‐sac larvae was significantly higher than those of the slow‐growing larvae, indicating that slow glowing larvae depended largely on rotifers rather than the yolk‐sac larvae.     

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.     

Carrying capacity and survival strategy for the Pacific bluefin tuna, Thunnus orientalis, in the Western Pacific

The carrying capacity for the Pacific bluefin tuna at each life stage is estimated and its survival strategy is examined numerically, using a new method to define the hypothetical capacity, the standard population, and the search volumes that are necessary and are feasible for the tuna. The carrying capacity for the adult is estimated at 1–2 × 10⁶ individuals, which corresponds with 5–10% of the hypothetical capacity and is comparable with the maximum levels of the southern and the Atlantic bluefin tuna populations. It is hypothesized semiquantitatively that the migration at each life stage and the remarkable decrement of growth at 120 days and about 40 cm occur as an evolutionary response to population excess over the carrying capacity. It is also hypothesized semiquantitatively that the early larvae have minimal food available in the Subtropical Water and develop the predatory morphology, high growth rate, and high mobility, however, at the expense of a high mortality as an evolutionary response to the tuna spawning in the Subtropical Water. This method may be an available tool to not only investigate the carrying capacity and survival strategy of a specific fish species, but also predict when and in how much abundance the fish species occurs in a specific area of its habitat.     

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.     

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.     

Reproductive biology of female Pacific bluefin tuna Thunnus orientalis from south-western North Pacific Ocean

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 incre ased with fork length; and (vii) preliminary estimates of spawning frequency between batches ranged 2–4.5 days based on analysis of postovulatory follicles.     

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.