Routine metabolic rate of southern bluefin tuna (Thunnus maccoyii)

Routine metabolic rate (RMR) was measured in fasting southern bluefin tuna, Thunnus maccoyii, the largest tuna species studied so far (body mass=19.6 kg (+/-1.9 SE)). Mean mass-specific RMR was 460 mg kg(-1) h(-1) (+/-34.9) at a mean water temperature of 19 degrees C. When evaluated southern bluefin tuna standard metabolic rate (SMR) is added to published values of other tuna species, there is a strong allometeric relationship with body mass (423 M(0.86), R(2)=0.97). This demonstrates that tuna interspecific SMR scale with respect to body mass similar to that of other active teleosts, but is approximately 4-fold higher. However, RMR (not SMR) is most appropriate in ram-ventilating species that are physiologically unable to achieve complete rest. Respiration was measured in a large (250,000 l) flexible polypropylene respirometer (mesocosm respirometer) that was deployed within a marine-farm sea cage for 29 days. Fasted fish were maintained within the respirometer up to 42 h while dissolved oxygen dropped by 0.056 (+/-0.004) mg l(-1) h(-1). Fish showed no obvious signs of stress. They swam at 1.1 (+/-0.1) fork lengths per second and several fed within the respirometer immediately after measurements.     

Routine metabolic rate of southern bluefin tuna (Thunnus maccoyii).

Routine metabolic rate (RMR) was measured in fasting southern bluefin tuna, Thunnus maccoyii, the largest tuna species studied so far (body mass=19.6 kg (+/-1.9 SE)). Mean mass-specific RMR was 460 mg kg(-1) h(-1) (+/-34.9) at a mean water temperature of 19 degrees C. When evaluated southern bluefin tuna standard metabolic rate (SMR) is added to published values of other tuna species, there is a strong allometeric relationship with body mass (423 M(0.86), R(2)=0.97). This demonstrates that tuna interspecific SMR scale with respect to body mass similar to that of other active teleosts, but is approximately 4-fold higher. However, RMR (not SMR) is most appropriate in ram-ventilating species that are physiologically unable to achieve complete rest. Respiration was measured in a large (250,000 l) flexible polypropylene respirometer (mesocosm respirometer) that was deployed within a marine-farm sea cage for 29 days. Fasted fish were maintained within the respirometer up to 42 h while dissolved oxygen dropped by 0.056 (+/-0.004) mg l(-1) h(-1). Fish showed no obvious signs of stress. They swam at 1.1 (+/-0.1) fork lengths per second and several fed within the respirometer immediately after measurements.     

Cloning and functional characterisation of a fatty acyl elongase from southern bluefin tuna (Thunnus maccoyii)

The synthesis of long chain polyunsaturated fatty acids (LCPUFA), such as eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), involves fatty acyl desaturase and elongase enzymes. The marine fish species southern bluefin tuna (SBT) can accumulate large quantities of omega-3 (n-3) LCPUFA in its flesh but their capacity to synthesize EPA and DHA is uncertain. A cDNA, sbtElovl5, encoding a putative fatty acyl elongase was amplified from SBT liver tissue. The cDNA included an open reading frame (ORF) encoding 294 amino acids. Sequence comparisons and phylogenetic analyses revealed a high level of sequence conservation between sbtElovl5 and fatty acyl elongase sequences from other fish species. Heterologous expression of the sbtElovl5 ORF in Saccharomyces cerevisiae confirmed that it encoded a fatty acyl elongase capable of elongating C_18/20 polyunsaturated fatty acid (PUFA) substrates, but not C₂₂ PUFA substrates. For the first time in an Elovl5, the substrate competition occurring in nature was investigated. Higher activity towards n-3 PUFA substrates than omega-6 (n-6) PUFA substrates was exhibited, regardless of substrate chain length. The sbtElovl5 preferentially elongated 18:4n-3 and 18:3n-6 rather than 20:5n-3 and 20:4n-6. The sbtElovl5 enzyme also elongated saturated and monounsaturated fatty acids.     

Isolation and partial characterisation of immunoglobulin from southern bluefin tuna Thunnus maccoyii Castelnau

Specific and total serum immunoglobulins were extracted by immunoaffinity, mannan-binding protein and Protein A affinity chromatography from southern bluefin tuna (Thunnus maccoyii Castelnau) immunised with rabbit IgG, and from non-immunised southern bluefin tuna. SDS-PAGE in 10% reducing gels revealed two heavy chains with molecular weights of approximately 74.6 +/- 1.3 kDa and 71.2 +/- 0.9 kDa, and two light chains with molecular weights of approximately 29 +/- 1.2 kDa and 28 +/- 1.0 kDa. Under non-reducing, but denaturing, conditions in 4% and 5% SDS-PAGE gels, a high molecular weight and a low molecular weight fraction were demonstrated. By gel filtration using Sephacryl HR 300 a molecular weight of 845 kDa, consistent with a tetramer, was obtained for the high molecular weight fraction, and a molecular weight of 168 kDa, consistent with a monomer, was obtained for the low molecular weight fraction. The extinction coefficient at A280 for the purified immunoglobulin (Ig) was determined to be 1.24. Tuna a-rabbit IgG Ig was reactive with all non-reduced mammalian IgG antigens tested, suggesting that common conformational antigenic determinants were recognised.     

Thermal effects on the blood respiratory properties of southern bluefin tuna, Thunnus maccoyii

Fasting-induced changes in fatty acid composition have been reported to occur within the body lipids of several types of animals; however, little is known about the changes in fatty acid profiles exhibited by reptiles subjected to prolonged fasting. This study characterizes the fatty acid profiles of six reptile species subjected to sublethal periods of fasting lasting 0, 56, 112, and 168 days. Analyses of fatty acid methyl esters (FAMEs) conducted on the total body lipids of rattlesnakes (Crotalus atrox), ratsnakes (Elaphe obsoleta), pythons (Python regius), boas (Boa constrictor), true vipers (Bitis gabonica), and monitor lizards (Varanus exanthematicus) revealed that all of the species exhibited similar characteristic changes in their fatty acid profiles during starvation stress. According to ANOVAs, the four most effective indicators of the onset of starvation were significant increases in the [1] fatty acid unsaturation index as well as ratios of [2] linoleic to palmitoleic acid, [3] oleic to palmitic, and [4] arachidonic to total fatty acid concentrations. The results of this study suggest that FAME analyses might be useful for identifying nutritional stress and/or starvation among squamate reptiles; however, forthcoming studies will be required to validate the generality of these responses. I also review the potential limitations of this approach, and suggest experiments that will be important for future applications of FAME analyses. Ultimately, it is hoped that FAME analyses can be used in conjunction with current practices as an additional tool to characterize the prevalence of starvation experienced by free-living reptiles.     

An epizootic of Caligus chiastos on farmed southern bluefin tuna Thunnus maccoyii off South Australia

In some years, large numbers of Caligus chiastos have been observed on the external surfaces of southern bluefin tuna, particularly on the head and eyes, in some sea cages in Spencer Gulf, Australia. As no epidemiological data were available, we monitored sea lice on tuna (N = 130) in 4 research cages sampled at 6 wk intervals during the 2005 farming season. No lice were observed on a sample of 10 wild-caught tuna when the cohort was transferred to cages in early April. By late May more than half the sampled tuna (22 of 40) were infected, with up to 42 parasites; we also recorded one unidentified Caligus sp. at this time. In early July the number of tuna infected with lice declined to 10%; in the final sample in late August none were detected. Prevalence in May was significantly higher than on other dates (p < or = 0.001), whereas mean abundances did not differ significantly (p > 0.05). The decline in prevalence corresponded with a seasonal fall in temperature, from ca. 17 degrees C in May to 14 degrees C in August. Counts of lice at the peak of infection were associated with the severity of eye damage (Spearman's rank correlation coefficient, r(S,38df) = 0.654, p < 0.001); this may be because lice graze on the cornea or because tuna injure their eyes when flashing (rubbing against objects). Counts at this time were also strongly and inversely correlated with the condition index (r(S,38df) = -0.707, p < 0.001). It appears that tuna become infested with adult sea lice via wild teleosts and elasmobranchs attracted to sea cages.     

Historical changes in juvenile southern bluefin tuna Thunnus maccoyii growth rates based on otolith measurements

Suspected historic changes in juvenile southern bluefin tuna Thunnus maccoyii growth rates were investigated using otolith increment width data. Four hundred and ninety otoliths were selected from fish estimated to be between 1 and 41 years-old. The distance between the first five annuli were measured on the otoliths, giving estimates of otolith growth for age classes 1+ to 4+ years for fish spawned from the early 1960s to mid 1990s. The data showed that growth rates of juveniles (age 1+ and 2+ years) started to increase at around 1979–1980, and that growth continued to increase throughout the 1980s and early 1990s. Lee's phenomenon was not observed in the data. Correlation tests did not reveal clear relationships between annual otolith growth and regional environmental variables such as sea surface temperature or Southern Oscillation Index. The increase in otolith growth, however, was consistent with juvenile growth estimates obtained from other sources, and correlated with large-scale trends in population size and environmental conditions.     

Age and growth rate determination of southern bluefin tuna, Thunnus maccoyii, using otolith banding

Otoliths of southern bluefin, Thunnus maccoyii , of between 42 cm and 167 cm f.l. , taken from waters off New South Wales, South Australia, and Western Australia were prepared to reveal annual banding. Methods of preparation and examination are detailed.
Otolith growth was demonstrated to be directly proportional to fish growth over the size range studied. Sampling over 13 months provided validation of the annual nature of bands for fish in their 3rd, 4th and 5th year of growth. Band formation of fish in their 2nd, 6th, 7th, 8th and 9th years of growth also appeared to be annular, though samples were available from an insufficient number of months for confident validation.
Von Bertalanffy growth parameters derived from determined age-at-length are L_L, 261.3cm; k , 0.108, t ₀, –0.157.     

Patterns of horizontal distribution of the larvae of southern bluefin (Thunnus maccoyii) and other tuna in the Indian Ocean

The fine- to coarse-scale distribution patterns of tuna larvaein the east Indian Ocean were investigated by a combinationof continuous transect sampling using surface tows and randomsampling using double oblique tows. Thunnus maccoyii was themost abundant species, reaching densities near patch centresof 22 m^–1 in surface tows, which is 25 times greater thanthe highest previous records for tuna larvae. Patches of T.maccoyiilarvae in areas of high abundance appeared to 5.2 for T.maccoyii.There was no change in the index when tow distance was doubledto 1200 m, older larvae. Lloyd's index of patchiness was consistentlyhigh for all tuna species, ranging from 3.0 to 5.2 for T.maccoyyii.There was no change in the index when tow distance was doubledto 1200 m, which suggests that the dominant patch size was somewhatlarger than the larger sampling interval. Sampling larvae atthe same site 4 days apart resulted in estimates of abundancethat differed by an order of magnitude. Abundance estimatedfrom a single station would depend largely on what day the stationwas occupied and where the sample was taken in relation to apatch. ¹Present address: Victoria Institute of Marine Sciences andDepartment of Zoology, University of Melbourne, Parkville, Victoria3052, Australia     

Amino acid composition and physico-chemical properties of bluefin tuna (Thunnus thynnus) myoglobin.

1. The heart ventricle myoglobin of Atlantic bluefin tuna has been purified and its amino acid composition has been determined. 2. The perturbing effect of guanidine hydrochloride on the molecular structure of tuna ferrimyoglobin and its corresponding apoprotein has been investigated by Soret absorbance and ultraviolet fluorescence. 3. The conformation-free energy of unfolding delta G0 has been calculated by thermodynamic treatments of the data concerning guanidine unfolding. 4. The results have been compared with other known myoglobins, particularly those of yellowfin tuna.     

Effects of immunostimulants on ranched southern bluefin tuna Thunnus maccoyii: immune response, health and performance

Ranched southern bluefin tuna Thunnus maccoyii were fed baitfishes supplemented with vitamins (predominantly E and C) or vitamins and immunostimulants, nucleotides and β-glucans, over 12 weeks after transfer and monitored for enhancement in immune response, health and performance through their 19 week grow-out period. Fish from two different tows were sampled separately at three different sampling points: at transfer to grow-out pontoons, at 8 weeks post-transfer and at harvest, 19 weeks post-transfer. Lysozyme activity was enhanced during vitamin supplementation compared to control fish. Performance (i.e. survival, condition index and crude fat), health (i.e. blood plasma variables including pH, osmolality, cortisol, lactate and glucose) and alternative complement activity were not commonly improved through diet supplementation. There were some tow-specific improvements in performance through vitamin supplementation including survival, selected parasite prevalence and intensity, and alternative complement activity. Immunostimulant supplementation also showed a tow-specific improvement in plasma cortisol level. Tow-specific responses may suggest that life history, previous health condition and husbandry can affect the success of vitamin and immunostimulant enhancement of immune response, health and performance of ranched T. maccoyii.     

Postprandial metabolic increment of southern bluefin tuna Thunnus maccoyii ingesting high or low‐lipid sardines Sardinops sagax

This study examined the postprandial metabolism and swimming speed of southern bluefin tuna Thunnus maccoyii when fed sardines Sardinops sagax of either high‐lipid and high‐energy content or low‐lipid and low‐energy content. Five groups of two or three T. maccoyii (mean ±s.e. mass = 19·8 ± 0·5 kg, n = 14) were fed either low [2·2% lipid, 5·5 MJ kg^?1 gross energy (GE)] or high‐lipid (12·9%, 9·2 MJ kg^?1 GE) S. sagax. Before feeding, T. maccoyii swam at 0·74 ± 0·03 body lengths s^?1 (n = 5) and their routine metabolic rate was 305 ± 15 mg kg^?1 h^?1. Swimming speed and metabolic rate of T. maccoyii increased following feeding. Thunnus maccoyii swam 1·3 and 1·8 times faster during digestion of low and high‐lipid S. sagax, respectively. Postprandial peak metabolic rate, duration of elevated metabolism and total postprandial metabolic increment were all greater for T. maccoyii that ingested high‐lipid S. sagax. When total postprandial increment is represented as a proportion of ingested energy, there was no difference between high and low‐lipid meals, equating to between 30 and 35% of ingested energy. It was estimated that increased postprandial swimming costs account for 25 and 46% of the total postprandial metabolic response for low and high‐lipid S. sagax meals, respectively. Specific dynamic action (SDA) accounts for c. 20% of ingested energy regardless of S. sagax lipid level. These results confirm that the postprandial metabolic increment of T. maccoyii is greater than most other fish species. Much of the high cost of postprandial metabolic increment can be attributed to increased postprandial swimming costs. For T. maccoyii, it appears that activity and SDA are not independent, which complicates bioenergetic evaluation. High postprandial metabolic costs accentuate the great energetic requirements of T. maccoyii.     

Purification and properties of rat liver mitochondrial glutathione peroxidase.

Glutathione peroxidase (glutathione:hydrogen peroxide oxidoreductase, EC 1.11.1.9) was purified from rat liver mitochondria. The enzyme was shown to be pure by polyacrylamide-gel electrophoresis and to contain multiple forms that differed in charge. Selenium was specifically associated with the enzyme. The enzyme was inhibited by iodoacetic acid and iodoacetamide in an unusual pattern of reduction by sulfhydryl compounds and pH dependency. The mitochondrial and cytoplasmic forms of the enzyme were compared, and an explanation of the inhibition patterns is offered.     

Trophic ecology of Atlantic bluefin tuna Thunnus thynnus larvae

Feeding intensity, diet composition, selectivity, energy ingestion and dietary niche breadth of larval Atlantic bluefin tuna Thunnus thynnus were studied on the eastern (Mediterranean) spawning grounds of the species. Larval T. thynnus were collected in the Balearic Archipelago (north-west Mediterranean Sea) during 2004 and 2005 using surveys specific for larval scombrids. Larvae between 2·6 and 8·7 mm standard length (L(S) ) are diurnal feeders, and 94% of the guts collected during daylight hours were full. The mean ±s.d. number of prey per gut was 7·1 ± 5·7, with mean ±s.d. ranging from 3·0 ± 1·6 in the smallest T. thynnus larvae to 11·1 ± 5·8 in 5·0-6·0 mm L(S) larvae. Up to 21 prey were found in a single larval gut (5·0-6·0 mm L(S) ) at the end of the day. Larvae progressively selected larger prey and exhibited increased carbon content concurrent with preflexion development of feeding and locomotory structures. Larvae of 5·0-6·0 mm L(S) exhibited positive selection of cladocerans over other prey (Chesson's index), whereas copepod nauplii dominated the diets of earlier stages. The dietary niche breadth measured increased initially but decreased at c. 5·5 mm L(S) . Appendicularians were found in the diet of larger larval sizes, but no piscivory was observed. Results are discussed in light of the sparse existing data for larval T. thynnus and other larval tuna species.     

Purification and characterization of a novel monomeric glutathione peroxidase from rat liver

A novel glutathione peroxidase, which is active toward hydroperoxides of phospholipid in the presence of a detergent, has been purified to homogeneity from a rat liver postmicrosomal supernatant fraction by ammonium sulfate fractionation and three different column chromatographies. From a DE52 column, glutathione peroxidase active toward phosphatidylcholine dilinoleoyl hydroperoxides was eluted in one major and two minor peaks. The enzyme in the major peak was found to be separated from the "classic" glutathione peroxidase and glutathione S-transferases and further purified by Sephacryl S-200 and Mono Q column chromatographies. The purified enzyme was found to be homogeneous on polyacrylamide gel electrophoresis under nondenaturing conditions as well as that in the presence of sodium dodecyl sulfate. The molecular weight of the enzyme as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 22,000, and that by gel filtration was comparable, indicating that the enzyme protein is a single polypeptide. The purified enzyme was found to catalyze the reduction of phosphatidylcholine dilinoleoyl hydroperoxides to the corresponding hydroxy derivatives. The isoelectric point of the enzyme was found at pH 6.2, and the optimum pH for the enzyme activity was 8.0. The enzyme was active toward cumene hydroperoxide, H2O2, and 1-monolinolein hydroperoxides in the absence of a detergent. The enzyme activity toward phospholipid hydroperoxides was minute in the absence of a detergent but was remarkably enhanced by the addition of a detergent. From these results, the presently purified enzyme is obviously different from the classic glutathione peroxidase and also from phospholipid hydroperoxide glutathione peroxidase purified from pig heart (Ursini, F., Maiorino, M., and Gregolin, C. (1985) Biochim. Biophys. Acta 839, 62-70), though considerably similar to the latter.     

A unique selenoprotein isolated from yellowfin tuna (Thunnus albacares) liver

A selenoprotein, with an approximate molecular weight of 2000, was isolated from yellowfin tuna (Thunnus albacares) liver. The selenium (Se) content of this selenoprotein fraction represented greater than 50% of the Se in the original liver extract. Most of the unrecovered Se was left in the pellet following homogenization. Although the protein was very sensitive to oxidizing conditions, it remained stable in the presence of reducing agents such as glutathione and dithiothreitol under a nitrogen atmosphere. After preparative isoelectric focusing of the purified selenoprotein, selenium was detected in three distinct bands, with the predominant band occurring at pH 6.2.     

Spatio-temporal variation in the elemental compositions of otoliths of southern bluefin tuna Thunnus maccoyii in the Indian Ocean and its ecological implication

The elements Na, Mg, Mn, Ca, Sr and Ba in otoliths of southern bluefin tuna Thunnus maccoyii , collected from their feeding ground in the central Indian Ocean and spawning ground between southern Java and north-western Australia were measured by laser-ablation inductively coupled plasma mass spectrometry (LA-ICPMS) and compared among sampling locations and developmental stages. The Na, Mg and Mn to Ca concentration ratios were significantly higher at the larval stage than at the adult stage, and the ratio reached a peak at the first inflection point of the otolith, mean ± s.d. 43·3 ± 4·9 days after hatching and decreased sharply to a low level thereafter. The temporal change of the elements:Ca ratios in the first inflection point corresponded to the life stage transition from larva to juvenile, indicating that the uptake rate of elements from ambient waters was significantly influenced by the ontogenetic change in the fish. The elemental composition at the otolith edge differed significantly in sub-adults on the feeding grounds and adults on the spawning grounds. Thus, the otolith elemental composition can be used as a biological tracer to study the time of the ontogenetic shift and to reconstruct the past migratory environmental history of T. maccoyii . In addition, the elemental composition of the otolith core of the adult was similar between feeding and spawning grounds, indicating that the fish in the Indian Ocean had the same larval origin, which is consistent with the single spawning population hypothesis.