Identification of growth‐related nucleotide polymorphism in cultured Pacific bluefin tuna,Thunnus orientalis

Pacific bluefin tuna (PBF), Thunnus orientalis, is commercially one of the most important species of tuna. In this study, amplified fragment length polymorphism (AFLP) screening was conducted to find the growth‐related polymorphic DNA in cultured PBF. Fish hatched in 2007 were harvested at an age of 818–1994 days. They were categorized into superior, average and inferior growth groups, depending on their growth score at the time of harvest. On AFLP screening of 24 fish, with eight fish from each group, 215 polymorphic DNA fragments were observed. A second amplification, with EcoRI + ACC and MseI + CCC primers, generated a polymorphic fragment of 630 bp at a rate of 80.0% (n = 15) in the superior, 56.3% (n = 16) in the average and 20.0% (n = 15) in the inferior growth groups. Polymerase chain reaction (PCR) primers, which could amplify both AFLP‐positive and AFLP‐negative loci, were developed using the consensus sequence outside the AFLP target fragment. Eleven haplotypes were obtained by sequence analysis of the PCR product at the AFLP target loci. Among those, haplotype 1 was statistically significant in the superior and average growth groups and could be used as a molecular marker for distinguishing the individuals with superior and average growth from those with inferior growth.     

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.     

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.     

Long‐term changes in recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) and environmental conditions around Japan

Recruitment of age‐0 Pacific bluefin tuna (Thunnus orientalis) from 1952 to 2014 was examined by a sequential regime shift detection method. The regime shifts in recruitment were detected in 1957, 1972, 1980, 1994 and 2009. The durations of regime shift ranged from 8–15 years and averaged 13.0 years. In both the total (1952–2014) and data rich (1980–2014) periods, negative relationships were found between recruitment and the Pacific Decadal Oscillation in autumn, and positive relationships were found between recruitment and sea surface temperature (SST) anomalies in the northern part of the East China Sea, in the southwestern part of the Sea of Japan, and in the waters off Shikoku and Tokai in summer and autumn. The 1994 and 2009 regime shifts in recruitment occurred in the same years as shifts in SST anomalies in the northern part of the East China Sea in summer. These results suggest that the ocean conditions in the northern part of the East China Sea are closely related to recruitment of Pacific bluefin tuna, and that the warmer conditions result in higher recruitment of the species.     

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.     

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.     

Heritability estimates for growth‐related traits in the Pacific oyster (Crassostrea gigas) using a molecular pedigree

Pacific oyster is one of the leading species in world aquaculture, but heritability estimation applying mixed‐family approach has not been actively pursued. In this study, heritability for growth‐related traits in the Pacific oyster was first estimated by creating a single cohort of 45 families in a full‐factorial mating design consisting of nine sires and five dams. A total of 270 offspring were analysed and parentage assignment inferred by six microsatellite markers achieved 100% success. All parents contributed to the spawn and a total of 42 full‐sib families were represented. Using an animal model, heritability estimates at 12 months of age were 0.49 ± 0.25 for shell height, 0.36 ± 0.19 for shell length, 0.45 ± 0.23 for shell width and 0.35 ± 0.17 for wet weight. Genetic correlation between shell height and wet weight was quite high (0.79 ± 0.25), suggesting that direct selection of shell height, which is an easily measurable trait, also improves wet weight. The results obtained in this study indicate that growth‐related traits could be improved by exploiting additive genetic effects through selective breeding.     

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.     

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.     

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.     

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.     

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.     

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.     

Identification of quantitative trait loci for growth‐related traits in the swimming crab Portunus trituberculatus

The swimming crab (Portunus trituberculatus) is an economically important species in Asian aquaculture. Implementing growth‐related traits of P. trituberculatus into genetic breeding programmes is an ongoing effort. We used a previously published genetic linkage map of P. trituberculatus, containing 55 simple sequence repeat (SSR) markers and 172 amplified fragment length polymorphism (AFLP) techniques to map quantitative trait loci (QTL) for growth‐related traits in a single full sibling F₂ family. Ten growth‐related traits were measured for QTL mapping. Composite interval mapping identified 9 QTL on the female map and 16 on the male map. Individual QTL with additive effects explained 11–38% of the phenotypic variance for various traits using the female parent's map, and from 1% to 21% using the male parent's map. Two QTL explaining a large percentage of variation in body weight were detected on chromosome 17 on the female map, and on chromosome 16 on the male map, and contributed 38% and 18% of the phenotypic variance respectively. This is the first study to report the detection and positioning of major QTL affecting growth in a true crab species (Brachyura). The mapping of growth‐related QTL in this study raises the possibility of improving the growth of P. trituberculatus through marker‐assisted selection.     

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.     

黄颡鱼遗传图谱构建及生长相关性状的QTL定位

以野生(♂)和人工养殖(♀)黄颡鱼杂交的100个F1个体为作图群体,用SSR、SRAP和TRAP3种DNA分子标记技术构建黄颡鱼的遗传连锁图谱。图谱整合了13个SSR标记,89个SRAP标记,26个TRAP标记。其中雌性框架图谱包括16个连锁群,图谱的长度为585.5cM;雄性框架图谱包括15个连锁群,图谱的长度为752.3cM;共享框架图谱包括5个连锁群,图谱的长度为231.3cM。用该连锁图谱对黄颡鱼的5个生长相关性状进行QTL扫描,在雌性图谱上检测到1个头宽的QTL,定位于第七连锁群上,LOD值为3.2,可解释的表型变异为13%。在雄性图谱上分别检测到1个体高和体长的QTL,均定位于第一连锁群上。体高QTL的LOD值为2.4,可解释的表型变异为12%。全长QTL的LOD值为2.1,可解释的表型变异为11%。3个QTL均可用于黄颡鱼的生长性状的标记辅助育种。