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.     

Development of visual cells in the Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

The development of rod and cone photoreceptor cells was investigated in the retinas of Pacific bluefin tuna larvae and juveniles, using RET-P1 monoclonal antibody labeling to identify photoreceptors. At 60 h after hatching, which was about when feeding began, opsin (presumably green opsin (Rh2)) was expressed in the outer segments of cone cells. At 15 days after hatching (dah), although many labeled cone cells were observed in the dorsal retina, the same type of cone cells had partially appeared in the ventral retina. The presence of rod cell bodies was confirmed by the expression of Rh1 opsin at 15 dah. At 21 dah, the presence of outer segments of rod cells was confirmed by the expression of Rh1 opsin and by morphology. The observations suggest that the cone cells were substantially operable upon the development of their outer segment at around the beginning of the post-larval stage, and the rod cells began to function at around 15 to 21 dah, before and during metamorphosis.     

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.     

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.     

Myxosporean and microsporidian infections in cultured Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> in Japan

During the parasitological survey of cultured juvenile Pacific bluefin tuna (PBT) Thunnus orientalis in 2007 and 2008, two myxosporeans and one microsporidian were found. Morphological and molecular analysis showed that the heart-infecting and brain-infecting myxosporeans are identified as Kudoa shiomitsui and K. yasunagai, respectively. This is a new host record for both species. High prevalence of infection (77–100%) with K. shiomitsui was observed in October to December (1–2 months post-transfer to sea cages), whereas only a few fish were infected with K. yasunagai. A microsporidian observed as white “cysts” in the trunk muscle of PBT had a resemblance to Microsporidium seriolae, the causative parasite of beko disease in yellowtail. Phylogenetic analysis showed that the microsporidian from PBT is closely related to but distinct from several other muscle-infecting species such as M. seriolae, Microsporidium sp. RSB, and Microsporidium sp. SH. Additionally, the spore dimension (2.7 × 1.5 μm on average) was remarkably smaller than the others. These results suggest that the present microsporidian is an undescribed species and designated Microsporidium sp. PBT. Prevalence and intensity of infection with Microsporidium sp. PBT reached a maximum of 100% and 20 cysts/fish, respectively. Although pathogenic effects of the two Kudoa species on fish health remain unknown, the microsporidian could be of concern to PBT aquaculture due to unsightly cysts in the musculature, reducing the market value of the fish.     

Development of phototaxis in the early life stages of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

We investigated the development of phototaxis in larval and early juvenile stages of Pacific bluefin tuna Thunnus orientalis to detail behavior development in relation to light. We observed the distribution of Pacific bluefin tuna in an illumination gradient tank from 1 to 26 days after hatching (DAH). Two long rectangular tanks (100 × 10 × 10 cm) were used for experiments; each tank was divided into four sections for observations and one tank was equally illuminated at 1.0 × 10³ lx as the control tank, and the other was at 4.0 × 10², 1.0 × 10³, 1.0 × 10⁴ and 7.0 × 10⁴ lx as the illumination gradient tank. Laboratory-reared larvae and juvenile were released into each tank and acclimatized for 10 min. Thereafter, the number of individuals in each section was counted for 5 times with 10-min intervals. Until 2 DAH (3.6 ± 0.1 mm BL), larvae distributed homogeneously in each section in both tanks. After 3 DAH (3.7 ± 0.1 mm BL) when the eyes of the larvae were pigmented, most fish moved into the 7.0 × 10⁴ lx section in the illumination gradient tank, while fish in the control tank maintained a uniform distribution during the experimental period (Chi-square test, P < 0.05). The results suggest that the post-larvae and juvenile stages show strong positive phototaxis.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Turbulence effect on survival and feeding of pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> larvae,on the basis of a rearing experiment

ABSTRACT:   Physical conditions such as oceanic turbulence related to food availability are considered to be important factors affecting fish larval survival. Rearing experiments were conducted to elucidate the effects of turbulence on the survival and feeding rates during the initial feeding period of Pacific bluefin tuna Thunnus orientalis . Six levels of turbulence intensity were provided by changing flow rates from pipes set on the bottom of rearing tanks. The result showed a dome-shaped relationship between turbulence level and survival rate, in which the feeding rate appeared higher at a logged turbulence energy dissipation rate of −6.32, and decreased at both higher and lower turbulence levels. Compared with the turbulence intensity in the ocean, the optimal turbulence level for Pacific bluefin tuna larvae corresponded to the turbulence caused by sea surface winds with speeds of 4–12.5 m/s. The estimated optimal turbulence intensity for Pacific bluefin tuna larvae is comparable to that for yellowfin tuna Thunnus albacares .     

Three cone opsin genes and cone cell arrangement in retina of juvenile Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis>

ABSTRACT:   In bluefin tuna aquaculture, collision of juveniles with the tank or net walls is a major cause of high mortality. This problem may be related to color sensibility of the visual mechanisms of this species. As a first step in understanding of color vision of Pacific bluefin tuna Thunnus orientalis , we applied a molecular technique and histology to study cone cell distribution in the retina of juvenile fish. We isolated three cone opsin genes encoding one blue-sensitive ( SWS2 ) and two green-sensitive ( RH2 ) visual pigments. In situ hybridization revealed that SWS2 mRNA is localized in the single-cone photoreceptors. The localization of the two RH2 signals in distinct cone cells was not determined, probably because of the high homology between the two RH2 genes. Single-cone photoreceptors appeared frequently in the ventral–temporal retina in approximately 80-mm fish and in the temporal retina in approximately 230-mm fish. These cone distributions may define a visual field with best color contrast vision in front and above the fish with a short wavelength (blue) reflecting target (sensed by single cones), and may be enhanced against the longer wavelength (green) background when fish see a target below them (sensed by double cones).     

Anesthesia induces stress in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>), Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) and Atlantic halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

Stress in response to anesthesia with benzocaine, MS-222, metomidate and isoeugenol was studied in Atlantic salmon (Salmo salar), Atlantic halibut (Hippoglossus hippoglossus), and Atlantic cod (Gadus morhua) with no concomitant stress from handling or confinement in association with anesthesia or sampling. All of the anesthetics tested induced a stress response in all species, displayed by a release of cortisol to the water. MS-222 anesthesia elicited the highest cortisol release rates, reaching maximum levels 0.5 h post-exposure and returning to basal levels after 3–4 h. Benzocaine anesthesia caused a bimodal response where the initial peak in cortisol release rate was followed by a second increase lasting towards the end of the trial (6 h). This bimodality was more profound in Atlantic salmon than in Atlantic halibut and Atlantic cod. Metomidate anesthesia induced the lowest release of cortisol of the agents tested in both Atlantic halibut and Atlantic cod, but resulted in a bimodal response in Atlantic salmon where the initial increase in cortisol release was followed by a larger increase peaking at 2–2.5 h post exposure before returning to basal levels after 5 h. The stress induced in Atlantic salmon by isoeugenol anesthesia resembled that of MS-222, but did not reach the same elevated level. Overall, the cortisol release was most profound in Atlantic salmon followed by Atlantic halibut and Atlantic cod.     

Proportions of Atlantic sailfish (<Emphasis Type="Italic">Istiophorus platypterus</Emphasis>) and spearfish (<Emphasis Type="Italic">Tetrapturus pfluegeri</Emphasis>) caught in tuna longline fisheries

The reliability of estimations of catches of pelagic longline fisheries is especially worrying in the case of non-target species such as the sailfish (Istiophorus platypterus) and spearfish (Tetrapturus pfluegeri), because they are often recorded aggregated in onboard maps. For this reason, generalized linear models were used to identify a number of important explanatory variables and their effect on sailfish and spearfish proportions. The response variable (number of sailfish in the aggregate capture) is assumed as binomial using the logit link function. The explanatory variables used were foram year, qrt (quarter), fleet, de (distance from equator) and d (distance from areas above sea level). The selected models showed that the equatorial and coastal regions presented higher densities or catchabilities of sailfish when compared to spearfish, as in the case for fleets of China Taipei and the United States. The lowest mean proportion of sailfish [sailfish/(sailfish + spearfish)] of the third quarter were driven mostly by the low estimations calculated for Japan and Brazil.     

Diet of young southern bluefin tuna <Emphasis Type="Italic">Thunnus maccoyii</Emphasis> in the southwestern coastal waters of Australia in summer

The diet of juvenile (predominantly age 1) southern bluefin tuna Thunnus maccoyii (SBT, N = 720), caught over 11 years of the recruitment monitoring survey off southern Western Australia during summer, consisted overwhelmingly of teleosts (97.4% by volume). Pilchard Sardinops sagax (27.4%V), blue mackerel Scomber australasicus (16.7%V), and jack mackerel Trachurus declivis (14.2%V) were the major taxa, with pilchard more abundant in coastal waters and jack mackerel more frequently encountered in fish caught closer to the shelf-edge. Prey size varied from 5 to 240 mm, with 67% of ingested items measuring between 30 and 50 mm. Pilchard dominated the prey size category 130–190 mm (84% by number), but the overall contribution of this species to the diet of juvenile SBT was much lower than previously reported. Future research in relation to the feeding ecology of juvenile SBT should focus on the biology and ecology of the young lifestages of the main prey species in this area and on prey distribution and dynamics as a key factor linking environmental change and SBT distribution.     

Recent expansion of Northeast Atlantic and Mediterranean populations of <Emphasis Type="Italic">Melicertus</Emphasis> (<Emphasis Type="Italic">Penaeus</Emphasis>) <Emphasis Type="Italic">kerathurus</Emphasis> (Crustacea: Decapoda)

We analysed the genetic diversity of Melicertus kerathurus (Penaeidae), a commercially valuable penaeid shrimp that is distributed in the Mediterranean Sea and eastern Atlantic Ocean. We examined the polymorphism of a 494 bp DNA segment of the mitochondrial COI region in 173 individuals, sampled in nine Mediterranean and two Atlantic samples, covering the whole range of the species from the tropical waters of the Gulf of Guinea to the eastern part of the Mediterranean Sea. The mean nucleotide and haplotype diversities were π = 0.00275 and h = 0.718, respectively, for the global data set, with the highest values occurring in the African samples and the lowest in the Adriatic Sea. A clear sample differentiation was found (F _st = 0.194), but this did not reflect a geographical pattern and there were only faint traces of an Atlantic–Mediterranean subdivision. Mismatch analysis and a high significant negative value of Tajima’s D suggested that M. kerathurus is not at mutation drift-equilibrium, but underwent a recent expansion after a period of low effective sample size. A postglacial recolonisation of the Mediterranean from an Atlantic refuge could be hypothesised based on these data.     

Onset of individual growth difference in larviculture of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> using fertilized eggs obtained from one female

We conducted a rearing experiment on Pacific bluefin tuna (PBT) larvae, which originated from one female broodstock, and determined the growth history of the larvae to clarify when the growth difference occurs. We investigated the otolith microstructure of the PBT larvae to examine the individual growth history and to predict the age and body size at the onset of individual growth difference. Since total length (TL) of fish can be back-calculated from otolith radius, we back-calculated TLs of 100 fish of 19 days post hatch (dph) divided into three size groups (small, intermediate, large). Growth difference was recognized from 3 dph (mouth opening), and the difference became larger thereafter. Growth of large-size fish was assumed to be promoted by the feeding conditions of PBT larvae.     

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.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.