Alterations in lipid metabolism and use of energy depots of gilthead sea bream (Sparus aurata) at low temperatures

Gilthead sea bream cultured along the northern Mediterranean coast are affected by the winter season when low temperatures reduce fish feed intake and growth. The coldest episodes can provoke a fish pathology known as ‘winter disease’. The effects of low temperatures, as well as concurrent fasting, have been studied by transferring three groups of gilthead sea bream from 16 °C to 14 °C, 12 °C and 8 °C. Fish at 12 °C and 8 °C refused food, whereas those at 14 °C were not fed following the temperature drop. Changes in body indices, organ composition, liver metabolism, and in particular, lipid fractions and their fatty acids were analysed on days 7 and 20 after the temperature shift. Only the rapid reduction of non-polar lipids in muscle was common for the three conditions. Fasting effects were linked to the maintenance temperature, being maximal after 20 days at 14 °C where fish body weight, hepatosomatic index, and perivisceral fat were reduced by 18%, 40%, and 60%, respectively. In this group, liver lipids did not change, as was the case for the enzymatic activities of liver glucose-6-phosphate and phosphogluconic acid dehydrogenases (G6PDH and PGADH) and lipoprotein lipase (LPL). In contrast, the liver of sea bream submitted to 8 °C accumulated large amounts of non-polar lipids (from 80 mg to 125 mg in 20 days), changing in size and aspect (bigger, pale, and friable). Simultaneously, liver LPL and hepatic lipase (HL) activities decreased. After 20 days at 8 °C, sea bream exhibited incipient acclimation responses to low temperatures: rising levels of unsaturation ratio in gill and liver polar lipids, of docosahexaenoic acid (DHA) in muscle polar lipids, and of G6PDH and PGADH hepatic activities.Fish at 12 °C presented some changes similar to those of the group at 14 °C (e.g., in morphological indices, and LPL and HL activities) and others like the group at 8 °C (increases in G6PDH and PGADH), suggesting a temperature threshold for gilthead sea bream (below 13 °C).     

Oxygen consumption and feeding rates of gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>) reveal lack of acclimation to cold

Low temperature has been implicated in inducing outbreaks of winter syndrome or winter disease in farmed gilthead sea bream (Sparus aurata). The responses of gilthead sea bream to reduced temperature followed by maintenance at low temperaturewere studied. In a first experiment, oxygen consumptionwas measured when water temperature was reduced from 18°C to 8 °C at either a rate of 1 °C· day-1 or as two sharp drops (from 18 °C to 12 °C, and from 12 °C to 8 °C). In a second experiment, the water temperature was reduced from 16 °C to 8 °C or 12 °C and then maintained for 20 days to study the fish acclimation to these temperatures. In both experiments, fish stopped feeding below 13 °C and did not resume feeding when maintained at low temperatures. The decrease in metabolic activity, expressed by the oxygen consumption rate, was directly related to the fall in water temperature: the Q10(18 °C-8 °C) values were between 2.2–2.5, independently of the descend rate in water temperature. However, we observed a more reduced metabolic rate when the water temperature was below 12 °C. Fish maintained at low temperatures showed only a partial recovery in oxygen consumption (15% at 8 °C and 20% at 12 °C) after 20 days. A higher metabolic rate together with a fasting-temperature condition meant that maintenance at 12 °C was more aggressive than at 8 °C, as revealed by the condition factor and energy needs. Data suggest that 12 .C could be a threshold temperature for the metabolic activity of gilthead sea bream. The relationship between low temperatures and their possible implication in the appearance of winter disease in gilthead sea bream is also discussed.     

Stress assessment,quality indicators and shelf life of three aquaculture important marine fish,in relation to harvest practices,water temperature and slaughter method

The objective of this study was to evaluate the effect of harvest practices and slaughter method on stress, quality and shelf life of whole fish (gilthead sea bream, European sea bass and red sea bream) towards the development of a humane slaughter practice. The use of hook and line resulted in significantly lower plasma cortisol and glucose in European sea bass and gilthead sea bream. Water temperature at harvest affected significantly the concentrations of stress indicators (plasma cortisol and glucose), mainly in European sea bass and gilthead sea bream. No significant differences were observed between the harvesting methods, regarding fish appearance, microbial spoilage and sensory scoring and subsequently shelf life. However, significantly lower hardness was recorded in fish harvested using the electric stunner, compared to the fish killed with slurry ice of the same harvesting period. Shelf life ranged between 13–18 days for gilthead seabream, 10–16 days for European sea bass and 10–14 days for red sea bream, depending on harvesting period.     

Correlation of changes in seasonal distribution and catch of red sea bream Pagrus major with winter temperature in the eastern Seto Inland Sea,Japan (1972–2010)

Red sea bream Pagrus major is a commercially important fish in Japan. In eastern Seto Inland Sea (SIS), the catch has increased from 297 tons in 1972 to 2,039 tons in 2010. We examined the relationship, 1972–2010, between increase in catch and winter temperature, based on the catch in February and March and the lowest water temperature at 10 m depth. In 1972–1986, the lowest water temperatures in the inner SIS areas (Osaka Bay, Harima‐nada, and Bisan‐seto) were <8°C, which is physiologically unfavorable for red sea bream. However, in 1987–2010 temperatures were generally ≥8°C. In the inner areas, the catch during winter had been minimal until the early 1980s, presumably because most red sea breams moved to the Kuroshio‐influenced (warmer) Kii Channel area. However, the winter catch in the inner areas of SIS increased from the late 1980s with warm winters. In addition, the catch between April and June, the spawning season, increased in the inner areas from the 1990s, and the catch rate of the inner areas was more than twice higher in the 2000s than in the 1980s. The results suggest that expansion of the distribution area during winter due to warm winter and increase in egg production in the inner areas greatly contributes to the increasing in catch in the eastern SIS.     

A drop in ambient temperature results in a transient reduction of interrenal ACTH responsiveness in the gilthead sea bream (Sparus aurata, L.)

This study investigated the effects of a drop in water temperature (18 °C to 9 °C in 24 h) on the pituitary and interrenal hormones of the gilthead sea bream Sparus aurata. The in vitro sensitivity of the interrenal tissue to ACTH, plasma levels of cortisol, ACTH, -MSH, GH, glucose, lactate and ions were determined. In vitro ACTH, stimulated the release of cortisol from isolated interrenal glands from control gilthead sea bream in a concentration dependent fashion. However, the interrenal cells were less sensitive to ACTH as soon as 24 h following the onset of the temperature drop. At this time, plasma cortisol and ACTH levels were raised, and plasma GH concentrations were decreased, whereas no significant changes were found in plasma -MSH. After 96 h plasma ACTH levels had recovered whereas plasma cortisol levels were still higher than controls after 8 days of the beginning of the experiment. Interrenal sensitivity had recovered after 8 days. The results may help to clarify the relationship between the stress response and the aetiology of the winter syndrome in sea bream.     

Effect of Storage Conditions on Total Volatile Base Nitrogen Determinations in Fish Muscle Extracts

Total volatile base nitrogen (TVBN) is a widely used fish spoilage indicator. Because TVBN is typically measured from muscle extracts, analytical laboratories would benefit from understanding how different conditions for storing extracts affect TVBN results. To examine this question, extracts from the muscle of gilthead sea bream (Sparus aurata) were prepared and stored. Different storage temperatures (4,??20, and??80°C) and storage periods (7 days to 2 months) were tested. To compare how well the different storage conditions worked for different TVBN levels, the study took advantage of the fact that TVBN values change depending on how long the gilthead sea bream is stored before extracts are prepared. Extracts were prepared from fish maintained in ice for 0, 4, and 10 days postmortem. Comparing the storage conditions tested, all offered statistically identical results independent of the storage period and temperature. Nevertheless, only the extract maintained at 4°C for 7 days achieved statistically equal values to not stored extracts.     

Temperature and dietary starch level affected protein but not starch digestibility in gilthead sea bream juveniles

A study was carried out with gilthead sea bream juveniles to assess the effect of water temperature (18 and 25°C) and dietary pregelatinized starch level (10, 20 and 30%) on digestibility of protein and starch and on the activity of proteolytic and amylolytic enzymes. ADC of pregelatinized starch was very high (>99%) irrespectively of dietary inclusion level, and it was not affected by water temperature. ADC of protein was also high (>90%) but improved at the higher water temperature. Dietary starch interacted with protein digestibility, which decreased as dietary starch level increased. Temperature affected both acid and basic protease activities, with acid protease activity being higher at 25°C and basic protease activity being higher at 18°C. However, total proteolytic activity and amylase activities were not affected by water temperature. Dietary carbohydrate exerted no effect on proteolytic or amylolitic activities. It is concluded that gilthead sea bream juveniles digest pregelatinized starch very efficiently irrespective of water temperature, due to adjustments of amylase activity to cope with temperature differences. Pregelatinized starch interacts negatively with protein digestibility, with the ADC of protein decreasing as dietary starch levels increase.     

Physiological stress and innate immune response in gilthead sea bream (<Emphasis Type="Italic">Sparus aurata)</Emphasis> and sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) exposed to combination of trimethoprim and sulfamethoxazole (TMP-SMX)

The purpose of this study was to determine whether gilthead sea bream and sea bass treated with combination of trimethoprim and sulfamethoxazole (TMP-SMX) differed in terms of physiological and innate immune biomarkers. Fish were exposed to TMP-SMX at 40 ppm concentration for 1 h as a prophylactic usage. Plasma cortisol, glucose, electrolytes (Ca, P, Na, K, Cl, and Mg) as well as plasma lysozyme activity, C-reactive protein (CRP), and ceruloplasmin (Cp) were measured soon after treatment and following 24 and 48 h in normal sea water for recovery. Treatment with TMP-SMX in both gilthead sea bream and sea bass led to an increase in plasma cortisol and glucose. Fluctuations in some electrolytes were found after treatment and during recovery period, however, the ratios of monovalent ions in treated sea bream were similar to control. Hematocrit values as well as plasma lysozyme activity in gilthead sea bream and sea bass were not affected by the treatment. CRP in gilthead sea bream and Cp in sea bass responded to the treatment with decreased levels. Both gilthead sea bream and sea bass displayed a rapid physiological stress response and sensitivity to TMP-SMX exposure, which requires more than 48-h period for regaining homeostasis.     

Effects of chronic confinement on physiological responses of juvenile gilthead sea bream, Sparus aurata L., to acute handling

Understanding how gilthead sea bream, Sparus aurata L., an important Mediterranean Sea species for aquaculture, respond physiologically to stressors commonly encountered in intensive rearing is important for effective production, as managing for stress is a major factor in maintaining healthy fish stocks. Our objective was to determine whether holding juvenile gilthead sea bream at a high density (HD), as a chronic stressor, would affect their physiological responses to a subsequent acute handling stressor. After acclimation at a low density (LD) of 6 kg m^?3 in 200‐L circular tanks containing 33–36 g L^?1 recirculating seawater at 19°C under a normal photoperiod, juvenile 37‐g gilthead sea bream were confined for 14 days at a HD of 26 kg m^?3 and then subjected to 30‐s aerial emersion in a dipnet. Plasma levels of cortisol, glucose, lactate, osmolality and chloride were determined in fish held in separate lots during LD (control) and HD confinement at 0, 1, 2, 7 and 14 days, and then after handling at 0, 1, 2, 4 and 8 h. Although plasma cortisol levels were similar in LD and HD fish groups after 14 d of confinement (15 and 23 ng mL^?1, respectively), the cortisol response in fish from the HD treatment at 1 and 2 h following acute handling (70 and 37 ng mL^?1, respectively) was only about half of that measured in the control group (139 and 102 ng mL^?1); plasma cortisol was similar in both groups by 4 and 8 h. In contrast, plasma glucose elevations in response to handling were higher at 4 and 8 h in the HD‐held fish (94 and 72 mg dL^?1, respectively) than in those from the LD treatment (59 and 51 mg dL^?1); glucose responses were similar in both groups at 1 and 2 h after handling and throughout confinement. Plasma lactate levels were higher in LD fish than in the HD group at the beginning of the experiment but were similar after 14 d confinement and responses to handling were similar (e.g. 33 and 35 mg dL^?1 at 1 h). Plasma osmolality showed increases during the first 2 h after acute handling but no differences were evident between the two density treatments at any time during confinement or posthandling. Plasma chloride levels did not change throughout the experiment. The reduced plasma cortisol response to acute handling likely resulted from negative feedback of mildly but chronically elevated circulating cortisol caused by the confinement stressor on the hypothalamic–pituitary–interrenal axis. While other post‐handling physiological changes also showed differences between treatment groups, the suppressed cortisol response in the HD‐held fish suggests a reduction in the gilthead sea bream's normal capacity to respond to an acute stressor.     

Effects of water temperature on mortality in Megalocytivirus‐infected rock bream Oplegnathus fasciatus (Temminck et Schlegel) and development of protective immunity

Rock bream iridovirus (RBIV) causes huge losses, especially in rock bream Oplegnathus fasciatus. Rock bream injected with RBIV and held at 29, 26, 23 or 20 °C had 100% mortality. Conversely, all infected fish held at 17 °C survived even after the temperature was progressively increased to 26 °C at 100 dpi. Rock bream exposed to virus and held for 2, 4 and 7 days at 23/26 °C before the temperature was reduced to 17 °C had mortality rates of 26.6/73.2%, 66.6/100% and 93.4/100%, respectively, through 100 dpi. When surviving fish had the water temperature increased from 17 to 26 °C at 100 dpi, they did not exhibit signs of disease and had low virus copy numbers (below 10³). To investigate the development of a protective immune, rock bream were infected with RBIV and held at 23 °C before shifting the water temperature to 17 °C at 4 dpi. All injected fish survived until 120 dpi. While 100% of the previously unexposed fish died, 80.2% of the previously infected fish survived. When the survivors were rechallenged again at 160 dpi, no further mortality occurred. The high survival rate of fish following rechallenge with RBIV indicates that protective immunity was established in the surviving rock bream.     

Incubation experimentale et developpement embryonnaire de la daurade royale, Sparus aurata (L.), a differentes temperaturesIncubation and embryonic development of gilthead bream, Sparus aurata (L.), at a range of temperatures

Eggs of raised gilthead bream, Sparus aurata (L.), were incubated to hatching at various temperatures ranging from 7.7°C to 26.3°C. For four stages of development, the relationship between temperature and incubation time is given. Time from fertilization to hatching varies from 135 h at 11°C to 40 h at 21.3°C. In our experiments no egg hatched below 11°C or above 22°C. The highest hatching rate and the lowest rate of larval abnormalities were both observed at 14.5°C which is also the spawning temperature.     

Olive oil by-product as a natural antioxidant in gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>) nutrition

The aim of the study reported here was to evaluate the efficiency of a natural antioxidant substance in gilthead sea bream (Sparus aurata) feeds. An olive oil by-product, olive mill vegetation water (VW), contains polyphenols, which have a strong antioxidant activity. A 147-day growth trial was conducted (monofactorial balanced, 4 × 3) with diet as the experimental factor. Two diets [isonitrogenous (crude protein 40%) and isoenergetic (gross energy = 18MJ kg⁻¹ on a dry weight basis] were formulated with 1 and 5% of VW (VW1, VW5); the control diet did not contain VW. A total of 600 juvenile gilthead sea breams (mean body weight 114.1 ± 5.7 g) were utilised. Production parameters and somatic indexes were calculated at the end of the growth trial. Antioxidant activity in fish fillets was investigated using TBARS and DPPH assays. A number of haematological parameters and digestive enzyme patterns were measured in fish in the middle and at the end of the experiment. The TBARS values showed slight delays in the development of oxidation in the fillet of fish fed with VW. There was no statistical difference between fish fed with the experimental diet and the control group, except for maltase activity, which increased with increasing VW in the feed. We found that the use of VW in a gilthead sea bream diet did not have any detrimental effects on gilthead sea bream production and physiological parameters and slightly improved the conservation of the fish fillets.     

Consumption of feeds containing the antibiotic sulfadiazine by gilthead sea bream (Sparus aurata,L.) and rainbow trout (Onchorynchus mykiss,Walbaum)

A method for studying the efficacy of sulfadiazine as a marker to estimate feed intake is presented. Feeding studies were carried out with two species, rainbow trout as a freshwater fish model and gilthead sea bream as a marine fish model, using two temperatures and two sizes of juveniles. The study showed the different feeding behaviour observed in both species, depending on the temperature and the number of days feeding the particles with sulfadiazine, and confirmed a dominant‐subordinate behaviour especially in the case of gilthead sea bream juveniles.     

Preliminary Results on Rearing of Sparus Aurata × Pagrus Pagrus Hybrids. Performance Comparison With the Parental Species

Hybridization between gilthead sea bream, Sparus aurata, and red porgy, Pagrus pagrus, was undertaken and viable hybrids were produced by an artificial hormonal-induced cross. The hybrids, from the cross of female red porgy and male gilthead sea bream (Pp × Sa), survived through the yolk-sac larval stage. The hybrids produced from the cross of female gilthead sea bream and male red porgy (Sa × Pp) presented higher mortality in the larval stage (98.4%) compared to gilthead sea bream (80.0%) and red porgy (92.5%). Afterwards, the Sa × Pp hybrid was compared to the offspring of its parental species under the same rearing conditions (tanks of 500 l capacity, natural light, ambient temperature, self-feeding with commercial feed) in a 40–week experiment. In this comparative study, the Sa × Pp hybrid had intermediate mortality rate (6%), compared to gilthead sea bream (0%) and red porgy (21%). The growth profile of this hybrid resembled the slow growing phases of its parents (red porgy in warm period and gilthead sea bream in cold period). The feed conversion ratio between fish forms was not significantly different (1.54–1.84 in the Sa × Pp hybrid, 1.53–1.56 in the maternal species and 1.33–1.40 in the paternal one). The body weight variation in the Sa × Pp hybrid was quite high (31.7–37.9%), compared to that in gilthead sea bream (19.7–24.7%) and red porgy (18.0–31.3%). With respect to body coloration, the Sa × Pp hybrid was distinguishable and classified into groups of red porgy-like (55.1%), gilthead sea bream-like (40.6%), and intermediate forms (4.3%). The preliminary results of this study proved that the produced Sa × Pp hybrid had too low a performance for aquaculture use, but the interesting consequent hereditary potential could be a useful tool in fish hybridization practice.     

Effects of diet supplementation with white tea and methionine on lipid metabolism of gilthead sea bream juveniles (Sparus aurata)

A growth trial was performed with gilthead sea bream juveniles (Sparus aurata) to evaluate the effect of diet supplementation with white tea and methionine on fish performance and lipid metabolism. For that purpose, four diets were formulated: a fish meal–based diet (Control) and diets identical to the control diet but supplemented with 2.9 % white tea (Tea), 0.3 % methionine (Met) or 2.9 % white tea plus 0.3 % methionine (Tea + Met). Growth performance and feed efficiency parameters, whole-body and liver composition, plasma metabolites concentration and liver glucose 6-phosphate dehydrogenase (G6PDH), malic enzyme (ME) and fatty acid synthetase (FAS) activities were determined. Feed intake was higher in fish fed methionine–supplemented diets, whereas this parameter and growth was decreased in fish fed white tea supplementation. Feed efficiency and protein efficiency ratio were not affected by diet composition. Plasma HDL cholesterol and total lipids concentration were higher in fish fed white tea–supplemented diets. Whole-body lipid, plasma glucose, liver glycogen concentration and liver G6PDH, ME and FAS activities were lower in fish fed white tea–supplemented diets. Results of the present study indicate that methionine seems to act as a feed attractant in diets for sea bream juveniles. Additionally, white tea is an important modulator of lipid metabolism in sea bream juveniles.     

The effects of a temperature rise on oxygen consumption and energy budget in gilthead sea bream

The rates of daily average, minimum and maximum oxygen consumption, specific dynamic action (SDA), ingestion and digestibility in gilthead sea bream, Sparus auratus L., were measured before, during and after a gradual rise of water temperature from 20 to 28 °C at a rate of 1 °C day-1. The relationship between energy gain and the metabolic costs has also been investigated. Oxygen consumption of juvenile sea bream had a clear daily rhythmicity, with low levels during darkness and higher levels during the light period. Both light and feeding processes raised the metabolism of sea bream, but fish fasted for 2 days also showed an increase in oxygen consumption during the light period. Acclimation to 28 °C was complete for the minimum oxygen consumption rate in 12 days, but daily average and maximum oxygen consumption rates on the 19th day after the change were still 33% higher than at 20 °C mainly due to increased SDA. The digestibility of food components did not change significantly throughout the experiment. These relationships are discussed with respect to the potential for growth when fish are reared at 28 °C, although the energy costs are high.     

Osmoregulation and differential penetration of two grey mullets, Chelon labrosus (Risso) and Liza ramada (Risso) in estuarine fish ponds

Grey mullet ponds situated in Arcachon Bay on the western Atlantic coast of France, display contrasting mixohaline habitats.The pattern of osmoregulation of Chelon labrosus and Liza ramada exhibiting differential penetration of fish ponds has been investigated. Key variables of salinity and temperature have been considered. The tolerances to low salinities and to fresh water have been determined. The dynamics of variation in plasma osmotic pressure and major plasma electrolytes (Na⁺, Cl^?, K⁺) were studied in terms of: (1) gradual adaptation to a decreasing salinity gradient; (2) effects of a long-term adaptation in fresh water; (3) effects of low temperatures on sea water adapted fish.Both species can adapt to a wide range of salinities; however, while L. ramada show a good hypoosmotic regulation, with no alteration in plasma concentration over a wide range of salinity and after a long stay (5 months) in fresh water, C. labrosus cannot live for long periods of time in a wholly fresh water habitat. Both field samples obtained from thick-lipped grey mullets forced to stay in a freshwater pond and experimentally adapted fish showed, after 3 months, an unequivoqual decrease in plasma sodium and chloride, and plasma osmolality decreased to the same extent; this pattern of no compensation is bound, sooner or later, to end in death. The inability of C. labrosus to develop a long-term hypoosmoregulation in fresh water is referred to salinity preferences which probably limit the upstream penetration of this species.Plasma osmolality and major electrolytes (sodium and chloride) increased in cold (4 °C) sea water. This response might be due to a reduced ability of grey mullets to osmoregulate in cold waters. In the poly-mesohaline ponds, Winter drop in temperature (4–6 °C) causes grey mullets to migrate to the meso-oligohaline ponds where the mean temperature is more elevated (8 °C) and temperature changes less rapid (6–9 °C).     

Growth and Survival of Larval Red Sea Bream Pagrus major and Japanese Flounder Paralichthys olivaceus Fed Microbound Diets

To establish the practical use of microbound diets (MBD) for larval fish in mass seedling production, rearing experiments of larval red sea bream, Pagrus major , and Japanese flounder, Paralichthys olivaeeus , were conducted. A mixture of various protein sources was used, and dietary amino acid patterns were approximated to those of larval whole body protein. Two thousand red sea bream larvae and 1,000 Japanese flounder larvae, all 10 days old, were placed in 100 liter tanks with running sea water under ambient water temperature, which ranged from 18 to 20C. The particle size of MBD was 125 μm at the beginning of the experiment and adjusted as fish size increased thereafter. Employing MBD together with a small amount of live food could sustain the growth and survival of larval red sea bream and Japanese flounder. Thus, data from the present study indicates that substitution of artificial feeds for live foods is possible for larval fish production, although improvements in MBD diets may be necessary before they are adequate for large scale seedling production.     

Effect of water temperature and light intensity on swim bladder inflation and growth of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> larvae

This study aimed to examine the effect of different water temperatures and light intensities on swim bladder inflation (SBI) and growth of red sea bream Pagrus major larvae to improve rearing techniques for this species. Two sets of experiments were conducted: different rearing temperatures were used in experiment 1 (19, 21, 23, and 25 °C), and different light intensities in experiment 2 (250, 1000, 4000, and 16,000 lx). Water temperature did not affect SBI frequency, but SBI initiation was accelerated at higher temperature, i.e., it was initiated on 3 days post-hatching (dph) at 25 °C and on 6 dph at 19 °C, suggesting that the promotion period for SBI, which needs a surface skimmer to be  run, also accelerated with increasing temperature in red sea bream larviculture. A higher temperature also significantly promoted larval growth, although the notochord of larvae at SBI initiation was shorter at higher temperatures. Light intensity had no effect on either the initiation or the frequency of SBI. However, light intensity of 250 lx significantly reduced early larval growth compared to light intensities higher than 1000 lx. These results indicate that light at an intensity greater than 1000 lx at the water surface is suitable for the early larviculture of red sea bream.