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

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

Comparison of the lipid profiles from wild caught eggs and unfed larvae of two scombroid fish: northern bluefin tuna (<Emphasis Type="Italic">Thunnus thynnus</Emphasis> L., 1758) and Atlantic bonito (<Emphasis Type="Italic">Sarda sarda</Emphasis> Bloch, 1793)

Lipids and essential fatty acids are determinants of the reproductive process in marine fish, affecting fecundity, egg quality, hatching performance, pigmentation and larval malformation. We have analyzed and characterized the lipids of eggs and unfed larvae of two wild caught scombroid fish, the Atlantic northern bluefin tuna (Thunnus thynnus) and Atlantic bonito (Sarda sarda). Dry matter and total lipid contents, polar and neutral lipid classes and total lipid fatty acid contents were determined in the eggs of bluefin tuna and eggs and unfed larvae during the development of Atlantic bonito. Bluefin tuna eggs had slightly but significantly more dry mass than bonito eggs but very similar lipid content. However, bluefin tuna eggs presented a higher polar lipid content due to increased proportions of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI). Bonito eggs and larvae showed increasing dry mass and decreasing lipid content with development. The proportion of polar lipids increased due to increased PE, PS and PI, whereas choline-containing polar lipids (phosphatidylcholine and sphingomyelin) remained relatively constant. Free cholesterol also increased, whereas the levels of other neutral lipids, especially triacylglycerol and steryl ester fractions, decreased, presumably due to utilization for energy to drive development. Bluefin tuna eggs had higher levels of n − 3 and n − 6 highly unsaturated fatty acids due to higher docosahexaenoic and arachidonic acid contents, respectively, than bonito eggs. The results are discussed in relation to the lipid and fatty acid requirements of larval scombroid fish in comparison to those of other larval marine finfish species under culture conditions.     

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

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

Molecular characterization of southern bluefin tuna myoglobin (<Emphasis Type="Italic">Thunnus maccoyii</Emphasis>)

The primary structure of southern bluefin tuna Thunnus maccoyii Mb has been elucidated by molecular cloning techniques. The cDNA of this tuna encoding Mb contained 776 nucleotides, with an open reading frame of 444 nucleotides encoding 147 amino acids. The nucleotide sequence of the coding region was identical to those of other bluefin tunas (T. thynnus and T. orientalis), thus giving the same amino acid sequences. Based on the deduced amino acid sequence, bioinformatic analysis was performed including phylogenic tree, hydropathy plot and homology modeling. In order to investigate the autoxidation profiles, the isolation of Mb was performed from the dark muscle. The water soluble fraction was subjected to ammonium sulfate fractionation (60–90 % saturation) followed by preparative gel electrophoresis. Autoxidation profiles of Mb were delineated at pH 5.6, 6.5 and 7.4 at temperature 37 °C. The autoxidation rate of tuna Mb was slightly higher than that of horse Mb at all pH examined. These results revealed that tuna myoglobin was unstable than that of horse Mb mainly at acidic pH.     

Environmental influences on Atlantic bluefin tuna (Thunnus thynnus) catch per unit effort in the southern Gulf of St. Lawrence

The Atlantic bluefin tuna (Thunnus thynnus) population in the western Atlantic supports substantial commercial and recreational fisheries. Despite quota establishment and management under the auspices of the International Commission for the Conservation of Atlantic Tunas, only small increases in population growth have been estimated. In contrast to other western bluefin tuna fisheries indices, contemporary estimates of catch per unit effort (CPUE) in the southern Gulf of St. Lawrence have increased rapidly and are at record highs. This area is characterized by the Cold Intermediate Layer (CIL) that is defined by waters <3°C and located at depths of 30–40 m in September. We investigated the influence of several in situ environmental variables on the bluefin tuna fishery CPUE using delta‐lognormal modelling and relatively extensive and consistent oceanographic survey data, as well as dockside monitoring and mandatory logbook data associated with the fishery. Although there is considerable spatial and temporal variation of water mass characteristics, the amount of available habitat in the southern Gulf of St. Lawrence (assuming a > 3°C thermal ambit) for bluefin tuna has been increasing. The percentage of the water column occupied by the CIL was a significant environmental variable in the standardization of CPUE estimates. There was also a negative relationship between the spatial extents of the CIL and the fishery. Properties of the CIL account for variation in the bluefin tuna CPUE and may be a factor in determining the amount of available feeding habitat for bluefin tuna in the southern Gulf of St. Lawrence.     

Prevalence,intensity and pathology of the nasal parasite Nasicola hogansi in Atlantic bluefin tuna (Thunnus thynnus)

Ectoparasitic flatworms of Nasicola (Monogenoidea: Capsalidae), which infect nasal epithelium of true tunas (Thunnus spp.), are not well studied, nor have their impacts on the host's olfactory organ been evaluated. Infections of Nasicola hogansi on Atlantic bluefin tuna, Thunnus thynnus, were investigated with emphasis on the relationship between infection prevalence, abundance and mean intensity with bluefin tuna size, sex, body condition and capture month, as well as histopathological effects. Commercially caught Atlantic bluefin tuna (n = 161, 185–305 cm curved fork length) from the Gulf of Maine were sampled during June through August 2009 for infections by N. hogansi. A total of 247 specimens of N. hogansi were collected, with a prevalence of 45.3%, mean abundance of 1.57 (CI: 1.21–2.03) and mean intensity of 3.45 (CI: 2.91–4.22). Neither fish sex nor landing month had a significant effect on parasite parameters. Larger and better-conditioned Atlantic bluefin tuna had a higher mean intensity of infection. Pathology associated with infection by N. hogansi included extensive necrosis, sloughing of the nasal epithelium and associated inflammation of underlying connective tissues. Further epidemiological and pathological study of this host–parasite system is warranted since impaired olfaction, if present, could adversely affect spawning and migration of this top ocean predator.     

Comparative intestinal absorption of amino acids in rainbow trout (Oncorhynchus mykiss), totoaba (Totoaba macdonaldi) and Pacific bluefin tuna (Thunnus orientalis)

The kinetics of amino acid absorption in the proximal section of three fish species were studied using an everted intestine technique and a pancreatic digest of casein (Tryptone), as the model amino acid mixture. Fresh intestine of rainbow trout (Oncorhynchus mykiss) was used to set up the experimental system, and results were compared with those obtained using intestines from totoaba (Totoaba macdonaldi) and bluefin tuna fish (Thunnus orientalis). The kinetics of amino acid absorption was sigmoidal for all amino acids and for each species of fish tested. In general, specific absorptions of essential amino acids were higher than those of non‐essential ones. No correlation between the concentration of amino acids in the Tryptone solution and the corresponding absorptions was found. The maximum specific absorption rates of all amino acids for trout were 10 times higher than those determined for totoaba and bluefin tuna. The relative amounts of the different amino acids preferentially absorbed in all three species were different. Results obtained from the everted technique may be applicable to the design of formulated diets for large fish species with commercial value.     

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

ABSTRACT:   In the present study, the cDNA encoding myoglobin (Mb) of bigeye tuna Thunnus obesus was cloned and its amino acid sequence deduced in order to investigate the relationship between the primary structure and thermostability of scombridae fish Mb. An open reading frame of bigeye tuna Mb cDNA contained 444 nucleotides encoding 147 amino acids. The primary structure of bigeye tuna Mb was highly conserved when compared with those of bluefin tuna and yellowfin tuna Mb, the sequence identity being 95.2–100.0%. It also showed relatively high identity (82.3–89.1%) with the counterparts of scombridae fish. Myoglobin was then isolated from the dark muscle of four scombridae fish including bigeye tuna. Differential scanning calorimetry and circular dichroism measurements on these Mb revealed that the thermostability of bigeye tuna Mb was lowest and that of skipjack Katsuwonus pelamis Mb highest among the scombridae fish Mb examined. The α-helical contents of scombridae fish Mb at 10°C were in the range of 39.8–44.8%, clearly lower than that of horse Mb (55.3%), suggesting instability of fish Mb. The melting temperatures of these Mb fell in the range of 75.7–79.9°C, lower than that of horse Mb (84.2°C). These results strongly suggest the instability of fish Mb.     

Morphological features and functions of bluefin tuna change with growth

Morphological features of Pacific bluefin tuna Thunnus orientalis have important functions for its fast swimming. Morphological features of tuna change with growth; therefore, morphological functions may develop during this process. In this study, we precisely quantified the morphology of bluefin tuna with growth from juvenile to young adult using a three-dimensional laser profiler and evaluated the fluid dynamic characteristics of tuna using computational fluid dynamics (CFD) and an accurate model based on measured data. As results of measurement of morphological features, the aspect ratio and sweepback angle, which are indices of hydrodynamic characteristics for a hydrofoil, suggested that the lift force of caudal fin was increased as the tuna grows. The results of CFD analysis showed that the coefficient of drag force gradually decreased with growth. Pectoral fins generated lift force, and the ratio of lift force to submerged weight (FL/SW) increased as the tuna grew to 0.2 m total length (TL). After the tuna exceeded 0.2 m TL, FL/SW changed to a wider range in angle of attack as the tuna grew. These results suggest that the morphological function of bluefin tuna develops to enhance its swimming ability as it grows from juvenile to young adult.     

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

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

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

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

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.     

美济礁附近海域3种金枪鱼肌肉成分检测与营养评价

为分析中国南海美济礁附近海域大目金枪鱼(Thunnus obesus)、蓝鳍金枪鱼(T.thynnus)、黄鳍金枪鱼(T.albacares)的营养成分与营养价值,该研究采用国标法检测了3种金枪鱼的水分、蛋白质、灰分、脂肪酸和氨基酸成分及其结构,并进行营养评价.结果显示,3种金枪鱼中黄鳍金枪鱼蛋白质含量最高,蓝鳍金枪鱼...     

Effects of open- and closed-system temperature changes on blood O2-binding characteristics of Atlantic bluefin tuna (Thunnus thynnus)

We investigated the effects of open- and closed-system temperature changes on the O₂ affinity of Atlantic bluefin tuna (Thunnus thynnus) blood using in vitro methods essentially identical to those previously employed on tropical tuna species. Bluefin tuna blood has a general O₂ affinity (P ₅₀ = 2.6–3.1 kPa or 19–23 mm Hg at 0.5% CO₂) similar to that of skipjack tuna, yellowfin tuna, and kawakawa blood (P ₅₀ = 2.8–3.1 kPa at 0.5% CO₂) but significantly above that of bigeye tuna blood (P ₅₀ = 1.6–2.0 kPa at 0.5% CO₂). We therefore hypothesize that bluefin tuna are less tolerant of hypoxia than bigeye tuna. Further, we found the P ₅₀ of bluefin tuna blood to be slightly reduced by a 10°C open-system temperature increase (e.g., from 4.83 kPa at 15°C to 3.95 kPa at 25°C) and to be completely unaffected by a 10°C closed-system temperature change. Bluefin tuna blood, therefore, had a significantly reduced Bohr effect when subjected to the inevitable changes in P CO ₂ and plasma pH that accompany closed-system temperature shifts (0.04–0.09 Δlog P₅₀ΔpH⁻¹) compared with the effects of changes in plasma pH accomplished by changing P CO ₂ alone (0.81–0.94 Δlog P₅₀ Δ pH⁻¹). This response is similar to that of skipjack tuna blood, but different from yellowfin or bigeye tuna blood. During closed-system temperature changes at oxygen levels above P ₅₀, however, bluefin tuna blood showed a reversed temperature effect (i.e., P O ₂ decreased in response to an increase in temperature). Unlike in other tuna species, temperature effects on O₂ affinity of bluefin tuna whole blood were similar to those previously reported for hemoglobin solutions, suggesting that red cell-mediated ligand changes are not involved.     

Tuna and swordfish catch in the U.S. northwest Atlantic longline fishery in relation to mesoscale eddies

To analyze the effects of mesoscale eddies, sea surface temperature (SST), and gear configuration on the catch of Atlantic bluefin (Thunnus thynnus), yellowfin (Thunnus albacares), and bigeye tuna (Thunnus obesus) and swordfish (Xiphias gladius) in the U.S. northwest Atlantic longline fishery, we constructed multivariate statistical models relating these variables to the catch of the four species in 62 121 longline hauls made between 1993 and 2005. During the same 13‐year period, 103 anticyclonic eddies and 269 cyclonic eddies were detected by our algorithm in the region 30–55°N, 30–80°W. Our results show that tuna and swordfish catches were associated with different eddy structures. Bluefin tuna catch was highest in anticyclonic eddies whereas yellowfin and bigeye tuna catches were highest in cyclonic eddies. Swordfish catch was found preferentially in regions outside of eddies. Our study confirms that the common practice of targeting tuna with day sets and swordfish with night sets is effective. In addition, bluefin tuna and swordfish catches responded to most of the variables we tested in the opposite directions. Bluefin tuna catch was negatively correlated with longitude and the number of light sticks used whereas swordfish catch was positively correlated with these two variables. We argue that overfishing of bluefin tuna can be alleviated and that swordfish can be targeted more efficiently by avoiding fishing in anticyclonic eddies and in near‐shore waters and using more light sticks and fishing at night in our study area, although further studies are needed to propose a solid oceanography‐based management plan for catch selection.     

Molecular characterization and expression analysis of heat shock proteins 40, 70 and 90 from kuruma shrimp <Emphasis Type="Italic">Marsupenaeus japonicus</Emphasis>

Heat shock proteins (HSPs) are proteins that are expressed more strongly when the cells are exposed to physiological and stressful conditions. In this study, the full-length cDNAs of heat shock proteins 40 (MjHSP40), 70 (MjHSP70) and 90 (MjHSP90) were cloned from kuruma shrimp Marsupenaeus japonicus. The open reading frames (ORFs) of the cDNA clones have lengths of 1,191, 1,959 and 2,172 bp and encode 396, 652 and 723 amino acid residues, respectively. The predicted MjHSP40 amino acid sequence contains a J domain, a glycine/phenylalanine-rich region, and a central domain containing four repeats of a CxxCxGxG motif, indicating that it is a type I HSP40 homolog. The signature sequences of the HSP70 and HSP90 gene families are conserved in the MjHSP70 and MjHSP90 amino acid sequences. The deduced amino acid sequences of MjHSP70 and MjHSP90 share high identity with previously reported shrimp HSP70s and HSP90s, respectively. The expression of MjHSP90 mRNA increased at 32°C. Additionally, the expressions of MjHSP40, MjHSP70 and MjHSP90 mRNAs increased in defense-related tissues (i.e., hemocytes and lymphoid organ) when the shrimp were challenged with white spot syndrome virus.     

Bluefin tuna (Thunnus thynnus) distribution in relation to sea surface temperature fronts in the Gulf of Maine (1994–96)

Fishery‐linked aerial surveys for bluefin tuna (Thunnus thynnus) were conducted in the Gulf of Maine (GOM) from July through October, 1994–96. Each year, from 507 to 890 surface schools were detected and their locations examined in relation to oceanographic conditions. Correlations between bluefin tuna presence and environmental variables were explored for sea surface temperature (SST), distance to a SST front, frontal density (relative density of all SST fronts seen in a given 1 km area for 2 weeks prior to each tuna sighting), and bottom depth and slope. Mean SST associated with bluefin schools was 18.1°C (±2.8). Schools were located at a mean distance of 19.7 km (±19.6) from SST fronts, and in water masses with an average frontal density of 28.2 m km^?2 (±35.7). Mean bottom depth of detected schools was 139.0 m (±70.3), and mean bottom slope was 0.7% rise (±0.7). A binomial generalized linear model fit to these variables indicated that bluefin are seen closer to fronts than locations in which no tuna were seen. Using simple and partial Mantel tests, we investigated the spatial correlation between bluefin tuna presence and the environmental variables, controlling for spatial autocorrelation. For each day that schools were sighted, we performed 24 Mantel tests, on a combination of response and predictor variables. The spatial relationship between bluefin tuna and SST fronts was inconsistent. Our analysis identified significant spatial structure in the bluefin school locations that had no significant correlation with any of the measured environmental features, suggesting that other untested features, such as prey density, may be important predictors of bluefin distribution in the GOM.     

Molecular evidence for cosmopolitan distribution of platyhelminth parasites of tunas (Thunnus spp.)

Global distribution of platyhelminth parasites and their host specificities are not well known. Our hypothesis was that platyhelminth parasites of large pelagic fishes are common around the world. We analysed molecular variation in three different taxa of platyhelminth parasites infecting four species of tunas: yellowfin tuna (Thunnus albacares, Scombridae) from Western Australia, southern bluefin tuna (Thunnus maccoyii, Scombridae) from South Australia, Pacific bluefin tuna (Thunnus orientalis, Scombridae) from Pacific Mexico and northern bluefin tuna (T. thynnus, Scombridae) from two localities in the Mediterranean (Spain and Croatia). Comparisons of ITS2 and partial 28S rDNA demonstrated two congeneric species of blood flukes (Digenea: Sanguinicolidae) from multiple hosts and localities: Cardicola forsteri from southern bluefin and northern bluefin tunas, and Cardicola sp. from Pacific bluefin and northern bluefin tunas; and a gill fluke, Hexostoma thynni (Polyopisthocotylea: Hexostomatidae), from yellowfin, southern bluefin and northern bluefin tunas. Partial 28S rDNA indicates that a second type of fluke on the gills, Capsala sp. (Monopisthocotylea: Capsalidae), occurs on both southern bluefin and Pacific bluefin tunas. This appears to be the first report of conspecific platyhelminth parasites of teleosts with a wide‐ranging geographical distribution that has been confirmed through molecular approaches. Given the brevity of the free‐living larval stage of both taxa of flukes on the gills (H. thynni and Capsala sp.), we conclude that the only feasible hypothesis for the cosmopolitan distribution of these flatworms is migrations of host tunas. Host migration also seems likely to be responsible for the widespread occurrence of the two species of blood flukes (Cardicola spp.), although it is also possible that these were translocated recently by the spread of infected intermediate hosts.