High‐quality spontaneous spawning in greater amberjack (Seriola dumerili,Risso 1810) and its comparison with GnRHa implants or injections

Production of sufficient high‐quality eggs of greater amberjack (Seriola dumerili) still constitutes the main bottleneck for commercial production of this species. The main objective of this study was to compare the quality of spontaneous spawn of greater amberjack with those obtained by either GnRHa injection or GnRHa implant protocols. Captive amberjack broodstock were distributed in three circular tanks of 40 m³. Broodstock from Tank 1 were not hormonally induced and spawned spontaneously, whereas those of Tank 2 were intramuscularly injected with GnRHa (20 µg/kg body weight) and those of Tank 3 were given EV‐500 µg GnRHa implants. The number of eggs per spawn obtained in the broodstock without hormonal treatment was larger than in those obtained with injections or implants. Egg quality was best in broodstock with spontaneous spawn, followed by GnRHa‐injected fish and then GnRHa implants. Besides, size of larvae from control and injected broodstock was similar between them and significantly higher (p < 0.01) than those from GnRHa implant spawn. Overall, this study showed that it is possible to obtain very high‐quality spontaneous spawn in greater amberjack, providing the adequate conditions. Furthermore, GnRHa weekly injections lead to similar egg viability and hatching rates than spontaneous spawn and higher fertilization rates than GnRHa hormonal implants, which is better than in previous studies.     

Positive phototaxis as the cause of jaw malformations in larval greater amberjack,Seriola dumerili (Risso, 1810): Mitigation by rearing in tanks with low‐brightness walls

Jaw malformations are a serious problem in fingerlings of the greater amberjack, Seriola dumerili. To establish a countermeasure against these malformations, we conducted a larval behavioural test to identify their root cause, and we investigated the ability of low‐brightness rearing tank walls to control their occurrence. Larval distribution was examined in a horizontally illuminated rectangular transparent aquarium (20 cm × 100 cm × 25 cm), and their distribution was biased towards the light source after this lighting was provided from 3 to 13 days post hatching (dph), indicating positive phototaxis; this response disappeared on 22 dph (there are no data between 13 and 22 dph). Larvae were observed to swim phototactically towards the light source and repeatedly collided with the tank wall (walling behaviour). To investigate the ability of wall brightness to control the incidence of jaw malformations, we compared their incidence in a rearing trial with different Munsell colour values for the tank walls: 2.11 (black), 5.85 (grey) and 9.52 (white). The incidence rate of jaw malformations at 25 dph was significantly lower in the tanks with lower‐brightness walls, that is, 5.8%, 22.5% and 26.8% for black, grey and white respectively. Larval walling behaviour was most intense for white, followed in order by grey and black. These results indicate that positive phototaxis elicits larval walling behaviour, causing jaw malformations, and that low‐brightness tank walls control this process.     

Effect of different rearing conditions on body lipid composition of greater amberjack broodstock (Seriola dumerili)

The aim of this study was to assess the effect of two rearing conditions: outdoor environment with great volume tanks (500 m³) and low stocking density (~0.4 kg m^?3); and indoor environment with smaller volume tanks (10 m³) and higher stocking density (~5 kg m^?3), on muscle, liver and ovary lipid composition of Seriola dumerili broodstock born in captivity. The rearing conditions tested seem to affect the pattern of lipid body deposition in broodstock fish of S. dumerili, increasing the muscle and liver triacylglycerides (TG) accumulation of fish kept under indoor conditions, probably due to reduced energy expenditure in swimming, with some variations in the fatty acid profile that may respond to the differences in the culture conditions tested. No significant differences were found for Gonadosomatic Index (GSI) or ovary lipid deposition between groups in this study, which may suggest that the conditions tested do not have a major effect on ovary development. However, one season later the females kept under outdoor conditions released eggs spontaneously, whereas those kept under indoor conditions did not spawn, suggesting that the conditions tested actually have an effect on the broodstock's reproductive fitness. More studies are needed in order to evaluate whether the conditions tested have or not any influence on ovary development.     

Effect of fish oil replacement and probiotic addition on growth,body composition and histological parameters of yellowtail (Seriola dumerili)

Fish (175 g of initial weight) were fed in triplicated groups with four diets formulated by 0% (FO 100), 75% (FO 25) and 100% (with and without probiotics, FO 0 and FO 0+) of fish oil replacement consisting of a mixture of linseed, sunflower and palm oils. After 109 days, growth and nutritional parameters were not affected by the treatment; however, fish fed with 0% of fish oil showed the lowest survival rate and without differences between the same diet with probiotics. As for biometric parameters, significant differences in the viscerosomatic index (VSI) were observed between fish fed the FO 0+ diet and the FO 100 and FO 25 diets. Results obtained from histological analysis did not detect inflammation in gut samples, while liver samples showed a remarkable steatosis in all four treatments. Total fish oil replacement produced a significant difference in the width of the lamina propria. The dietary inclusion of probiotics in the FO 0+ diet seems to favour a recovery of intestine histology. In addition, as fish oil substitution increased, the width of the lamina propria also increased. In conclusion, it is possible to affirm that the four diets administrated to Seriola dumerili did not compromise the correct development of the animals.     

Effect of different algae used for enrichment of rotifers on growth, survival, and swim bladder inflation of larval amberjack Seriola dumerili

The effect of algae with different DHA contents used for the enrichment of rotifers on the growth performance, survival, and swim bladder inflation of larval amberjack Seriola dumerili was investigated. Rotifers were enriched with freshwater Chlorella vulgaris containing three levels of DHA (rotifer containing DHA 0.04, 0.60, 1.32?g DHA 100?g^?1 DM) and Nannochloropsis (0.04?g DHA 100?g^?1 DM; 2.54?g EPA 100?g^?1DM). The larvae were fed the enriched rotifers in triplicate from 3?days post-hatch for 7?days in static condition. The same algae used for rotifer enrichment were supplied to the larval tanks. Growth and survival rate of fish fed the rotifers enriched with Nannochloropsis were higher than those of fish fed the rotifers enriched with all three Chlorella treatments. Swim bladder inflation was lowest in fish fed the rotifers enriched with Nannochloropsis. The proportion of EPA was higher in fish fed the rotifers enriched with Nannochloropsis, while that of DHA increased proportionally with the DHA levels in the rotifers enriched with Chlorella. These results suggest that rotifers enriched with Nannochloropsis (EPA-rich rotifers) are effective to enhance growth and survival, but DHA instead of EPA is essential to improve the swim bladder inflation in amberjack larvae.     

Encephalomyelitis associated with microsporidian infection in farmed greater amberjack, Seriola dumerili (Risso)

An outbreak of a disease characterized by a peculiar spiral movement in farmed greater amberjack, Seriola dumerili (Risso), occurred in Kagoshima Prefecture, Japan, in May 2008, immediately after importing the fish from China. Although neither bacteria nor viruses were detected in routine diagnostic tests, histopathological observations of the affected fish revealed severe inflammation in the tegmentum of the brain including the medulla oblongata and the anterior part of the spinal cord. In addition, a microsporidian parasite was observed in the nerve cell bodies or axons in the inflamed tissues. We identified a microsporidian small subunit rRNA gene (SSU rDNA) from the lesion, and the sequence showed 96.1% identity with that of Spraguea lophii. Subsequent in situ hybridization using probes presumably specific to the SSU rRNA confirmed that the parasite observed in histopathology harboured the identified SSU rRNA. Apparently degenerated microsporidian cells or spores were also frequently observed in tissue sections. Thus, the disease was most probably caused by the infection of a hitherto unknown microsporidian parasite that has a genetic affinity to the genus Spraguea, in the central nervous system of the amberjack.     

Effect of temperature on growth performance of greater amberjack (SERIOLA DUMERILI Risso 1810) Juveniles

In order to successfully diversify Mediterranean aquaculture, it is necessary to determine optimum culture conditions of potential candidate species such as greater amberjack (Seriola dumerili). Among culture conditions, rearing temperature is a key factor for achieving optimum growth and maintaining fish welfare. However, little is known about the optimum culture conditions of greater amberjack (Seriola dumerili). Thus, the aim of this study was to determine the effect of three different rearing temperatures (17, 22 and 26°C) during 120 days on growth performance, body morphometry, biochemical composition, gut transit and liver morphology of greater amberjack (Seriola dumerili) juveniles. After 120 days of rearing, fish raised at 26°C showed higher (p < .05) body weight and specific growth rate than fish held at lower temperatures, as well as improved feed utilization, protein efficiency and nutrient retention percentages. Fish stomach emptying was faster (p < .05) in fish raised at 26°C than in fish held at 22°C and 17°C. Similar results were obtained for gut transit time, being gut emptying faster (p < .05) in fish reared at 26°C than in fish cultured at lower temperatures. Rearing temperature also induced changes in fish morphology which resulted in a higher (p < .05) caudal propulsion efficiency index for fish reared at 26°C. Based on these results, we conclude that greater amberjack fingerlings perform better at 26°C than at 22°C or 17°C.     

Partial and total replacement of fishmeal by a blend of animal and plant proteins in diets for Seriola dumerili: Effects on performance and nutrient efficiency

A 154‐day trial was performed to assess the use of an alternative protein blend (corn gluten, krill and meat meal) as a substitute for fishmeal in diets for juvenile yellowtail, using four isolipidic (140 g/kg) and isoenergetic diets (24 MJ/kg) with the same digestible protein content (50%). The control diet was FM100, without replacement, and in FM66, FM33 and FM0, fishmeal was replaced at 33 g/kg, 66 g/kg and 100 g/kg, respectively. At the end of the experiment, no differences in growth parameters were observed. Fish fed the FM0 diet exhibited the lowest survival (23%). This high mortality may be due to different factors, such as a dietary amino acid imbalance or some antinutrient factors contained in the alternative ingredients. Feed intake, feed conversion ratio, digestible protein intake and protein efficiency ratio were similar in all diets. However, digestible energy intake and protein efficiency retention were lowest in fish fed the FM0 diet. Apparent digestibility coefficients for protein, energy and amino acids diminished as a substitution for fishmeal increased. Significant differences were observed in the diet whole‐fish body profile amino acid retention (AAR) ratio for the seven essential amino acids. In summary, total fishmeal replacement by the blend assayed was not feasible for yellowtail. The FM66 diet resulted in good growth, high survival and good nutrient efficiency.     

Effects of dietary docosahexaenoic acid on growth performance,fatty acid profile and lipogenesis of blunt snout bream (Megalobrama amblycephala)

Six diets were designed to investigate the effects of dietary docosahexaenoic acid (22:6n‐3; DHA) levels (0.5, 1.3, 2.3, 4.2, 8.1 and 15.9 g/kg diets) on growth performance, fatty acid profile and expression of some lipogenesis‐related genes of blunt snout bream (Megalobrama amblycephala). Fish (average weight: 26.40 ± 0.11 g) were randomly fed one of six diets for 8 weeks. Results indicated that the final body weight (FBW) and specific growth rate (SGR) of fish fed 1.3 g/kg DHA were significantly higher than other groups except for the 2.3 g/kg DHA (p < .05). Compared with other groups, the number of lipid droplet clusters of the liver stained with oil red O in the 2.3 g/kg DHA group was the highest, which was consistent with the lipid contents of whole body and liver. The DHA proportion in liver and muscle significantly increased with the increasing dietary DHA levels (p < .05), which reflected fatty acid profiles of diets. The highest mRNA expressions of acetyl‐CoA carboxylase α (ACCα), fatty acid synthase (FAS) and sterol regulatory element‐binding protein‐1 (SREBP‐1) occurred in the 1.3 g/kg DHA group, followed by 2.3 g/kg DHA. In summary, the supplementation of 1.3–2.3 g/kg DHA could improve growth performance and lipogenesis, and the dietary DHA could improve DHA and PUFA proportion in liver and muscle.     

Effect of feeding rotifers enriched with taurine on the growth and survival of larval amberjack Seriola dumerili

The effect of feeding rotifers enriched with taurine on the growth performance and survival of larval amberjack Seriola dumerili was investigated. Rotifers were enriched with a commercial taurine supplement at four levels (0, 200, 400, and 800 mg/l). The larvae were fed the enriched rotifers in triplicate from 3 days post-hatch for 7 days under static conditions. The average taurine contents of the rotifers were 1.5, 2.7, 4.2, and 7.2 mg/g dry matter, respectively. The growth of the fish fed rotifers enriched with the taurine supplement at 800 mg/l was significantly (P < 0.05) improved compared with that of the fish fed the rotifers without taurine enrichment. The survival rate improved proportionally up to a taurine supplement level of 400 mg/l, but no significant differences in survival were observed among treatments. The fraction of the larvae with inflated swim bladders did not vary significantly between treatments. Taurine content in the whole fish body increased with the taurine level in the rotifers. These results suggest that taurine enrichment of rotifers is an effective method of enhancing the growth of amberjack larvae.     

Nutritional evaluation of live food organisms and commercial dry feeds used for seed production of amberjack Seriola dumerili

To improve the nutritional quality of live foods and dry feeds ordinarily used for the seed production of amberjack Seriola dumerili, the nutrient contents of rotifers, Artemia nauplii and commercial feeds used in two larval production stations were evaluated. For comparison of the nutrient contents, artificially produced larvae, wild-caught juveniles and wild zooplankton samples were also analyzed. The proportions of 22∶6n-3 in the polar lipid of the cultured larvae increased by feeding the dry feeds. The taurine contents of the cultured larvae reflected the contents of their foods (rotifers<dry feed<Artemia nauplii). The taurine content and the proportion of 22∶6n-3 in Acartia spp. were higher than in foods fed to the larvae. These parameters in the wild juveniles were higher than the cultured ones. The A/E ratios [(each essential amino acid/total essential amino acids)×1000] of the total amino acids of the live foods and dry feeds were similar to those of the cultured larvae, except for the lower ratios of histidine, arginine, threonine and lysine in the live foods. The mucosal folds of the intestine of the cultured larvae did not show typical signs of dietary phospholipid deficiency. These results suggest that requirements of nutrients such as 22∶6n-3 and taurine should be determined for mass production of amberjack seeds.     

Effect of increasing docosahexaenoic acid content in weaning diets on survival,growth and skeletal anomalies of longfin yellowtail (Seriola rivoliana,Valenciennes 1833)

Five isoproteic (54.8%) and isolipidic (24.1%) microdiets, which varied in their docosahexaenoic acid (DHA) content (0.25%, 0.75%, 1.64%, 1.99% and 3.17%; dw), were manufactured to determine its effects on longfin yellowtail Seriola rivoliana larvae in terms of fish biological performance, whole body fatty acid profile and incidence of skeletal anomalies from 30 dah (11.31 ± 1.79 Total Length, TL) to 50 dah (19.80 ± 0.58 mm TL). The inclusion of dietary DHA up to 3.17% (dw) improved larval resistance to air exposure, although DHA did not significantly affect fish final growth or final survival. Indeed, high levels of dietary DHA (1.99% and 3.17%, dw) tended to increase the incidence of skeletal anomalies in S. rivoliana larvae, albeit no significant differences were observed. Furthermore, the occurrence of severe anomalies such as kyphosis and lordosis, was mainly associated to the larvae fed the highest levels of dietary DHA. In terms of survival, increasing dietary DHA levels did not significantly affect longfin yellowtail survival rate, despite a tendency for enhanced survival. The results of the present study proved that the inclusion of dietary DHA in inert diets up to a 3.17% (dw) and a DHA/EPA ratio above 3.1 increased the final survival and stress resistance in S. rivoliana larvae.     

Dietary docosahexaenoic acid to eicosapentaenoic acid (DHA/EPA) ratio influenced growth performance,immune response,stress resistance and tissue fatty acid composition of juvenile Japanese seabass,Lateolabrax japonicus (Cuvier)

A 10‐week feeding experiment was conducted to investigate the effects of dietary DHA/EPA ratio on juvenile Japanese seabass reared in sea floating cages. Six practical diets were formulated differing only in DHA/EPA ratio: 0.55 (Diet D/E0.55), 1.04 (D/E1.04), 1.53 (D/E1.53), 2.08 (D/E2.08), 2.44 (D/E2.44) and 2.93 (D/E2.93). All diets had the same contents of total n‐3 long‐chain polyunsaturated fatty acid (LC‐PUFA) and arachidonic acid (ARA). The results showed that the final weight and specific growth rate significantly increased with increasing dietary DHA/EPA ratio from 0.55 to 2.08 and thereafter declined. Activities of lysozyme and superoxide dismutase in serum in groups with DHA/EPA of 1.53–2.93 was significantly higher compared to group D/E0.55 while the activity of serum alternative complement pathway in group D/E2.93 was significantly lower compared with group D/E1.53. However, no difference was observed in activities of both respiratory burst of head kidney macrophage and serum catalase among dietary treatments. The per cent survival after air exposure in group D/E1.53, D/E2.08 and D/E2.93 was significantly higher compared with group D/E0.55. The fatty acid composition of whole body and tissues reflected closely those of diets, while fish accumulated more DHA than EPA in fish tissues, especially in muscle. These results suggested that at the same dietary contents of n‐3 LC‐PUFA (appr. 18% of TFA) and ARA (appr. 0.7% of TFA), moderate dietary DHA/EPA ratios of 1.53–2.08 significantly enhanced growth performances, certain innate immune responses, and the stress tolerance of Japanese seabass, in accordance with the preferential incorporation of DHA into fish tissues.     

Increased melanomacrophage centres in the liver of reproductively dysfunctional female greater amberjack Seriola dumerili (Risso, 1810)

The greater amberjack Seriola dumerili is a new aquaculture fish that may display reproductive dysfunctions. During extensive follicular atresia, which is a common reproductive dysfunction in females during vitellogenesis, part of the reabsorbed yolk returns to the liver to be metabolized and recycled. Melanomacrophage centres (MMCs) are aggregates of macrophage-like cells that play a role in the destruction, detoxification and recycling of endogenous and exogenous materials, and have been associated with systemic stress. Wild and captive-reared greater amberjack were sampled in the Mediterranean Sea during two different phases of the reproductive cycle. The liver of reproductively dysfunctional captive-reared females sampled during the spawning season showed a high density of both MMCs and apoptotic cells. A weak liver anti-cytochrome P450 monooxygenase 1A immunoreactivity was observed, suggesting that the examined fish were not exposed to environmental pollutants. We propose that the observed increase in MMCs and apoptosis in captive-reared fish was related to the hepatic overload associated to the metabolism of yolk proteins reabsorbed during extensive follicular atresia. Since follicular atresia is a frequent physiological and pathological event in teleosts, we suggest that the reproductive state should be always assessed when MMCs are used as markers of exposure to stress or pollutants.     

Effect of Continuous Food Supply on the Oxygen Consumption of Young Mediterranean Yellowtail (Seriola dumerili Risso, 1810)

The aim of this work was to investigate the effect of a continuous food supply on the oxygen consumption of young Mediterranean yellowtails (Seriola dumerili Risso, 1810) during the photophase period (14LL:10DD). Four groups with six fish each, weighing individually approx. 600 g, were distributed into open 2 m³ quadrangular fibreglass tanks, and fed with commercial pellets for 60 days. Two of the tanks received a continuous feed supply by means of automatic band feeders (ABF), whilst fish in the other two tanks were fed by hand, twice a day. To improve accuracy in calculating oxygen consumption, the effect of oxygen diffusion between the air and the water was determined by the use of a control tank of similar characteristics, containing no fish. Oxygen consumption instantaneous rate was calculated by the difference between dissolved oxygen concentrations in tanks with and without fish, multiplied by the water flow and divided by the estimated total fish biomass in each tank. In both feeding regimes, an increase in the oxygen consumption levels after feeding commencement was observed. The amplitude, however, was lower and the duration of feeding effect was higher in the group fed by ABF.     

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

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

Visualization of musculature of the larval greater amberjack Seriola dumerili (Risso) using whole‐mount immunostaining with special reference to cranial muscles

Muscles play important roles in feeding, respiration and swimming during not only adult stage but also larval stage in fish. However, there is no information of the muscle development in larval greater amberjack, Seriola dumerili (Risso). Here, we investigated muscle development in the greater amberjack, focusing primarily on the cranial muscles from 0 days post hatch (dph) to 12 dph using a modified whole‐mount immunohistochemical staining method. We found that the muscles required for feeding develop by 3 dph, when the larvae begin to feed. Subsequently, muscle composition in the dorsal branchial arches changes to the adult form between 5 and 8 dph. At 8 dph, all the muscles required for feeding and respiration appear, whereas the dorsal, pelvic and caudal fin muscles required for swimming develop later. This report provides fundamental information on larval greater amberjack muscle development, which will enable the detection of abnormal larval muscles and improve larval rearing techniques by modifying environmental conditions.