Establishment and characterization of India's first marine fish cell line (SISK) from the kidney of sea bass (Lates calcarifer)

A continuous cell line (SISK) from kidney of sea bass, Lates calcarifer, has been established and characterized. The cell line was maintained in Leibovitz' L-15 supplemented with 15% fetal bovine serum. This cell line has been subcultured more than 100 times over a period of 2 years. The SISK cell line consists of predominantly of epithelial-like cells. These cells showed strong positive for epithelial markers such as cytokeratin 19 and pancytokeratin. The cells were able to grow at temperature between 25 and 32 °C with optimum temperature of 28 °C. The growth rate of sea bass kidney cells increased as the FBS proportion increased from 2% to 20% at 28 °C with optimum growth at the concentrations of 15% or 20% FBS. The distribution of chromosome number was 30 to 56 with a modal peak at 48 chromosomes. Polymerase chain reaction products were obtained from SISK cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this was found to be susceptible to MABV NC1 and nodavirus, and the infection was confirmed by RT-PCR and CPE. This suggests that the SISK cell line has good potential for the isolation of various fish viruses. This cell line has been shown to be susceptible to bacterial extracellular products. The SISK cell line is the India's first marine fish cell line.

Post-haemorrhagic anaemia in sea bass, Dicentrarchus labrax (L.), caused by blood feeding of Ceratothoa oestroides (Isopoda: Cymothoidae)

The effects of the fish parasitic isopod, Ceratothoa oestroides (Risso), on haematological parameters of its cage-cultured sea bass host, Dicentrarchus labrax (L.), were studied. Analyses of blood parameters (cell counts, haemoglobin content and haematocrit) were carried out on parasitized and unparasitized sea bass from a fish farm in Turkey. Parasitized fish had significantly lowered erythrocyte counts, haematocrit and haemoglobin values and significantly increased leucocyte counts. Blood feeding by C. oestroides thus produces a post-haemorrhagic anaemia and the fish appear to mount an immune response to the presence of parasites.

Potential of cottonseed oil as fish oil replacer in European Sea Bass feed formulation

Triplicate groups of 20 European sea bass (35 g) were fed five diets in which the added lipid was 100% fish oil (FO), 40% (CSO40), 60% (CSO60), 80% (CSO80) and 100% (CSO100) refined cottonseed oil (CSO), for a period of 120 days. Overall fish growth, feed conversion ratio and protein utilization were unaffected by dietary treatment, but hepatosomatic and visceral fat indexes increased with increasing dietary CSO. Fillet fatty acid composition of total lipids reflected the fatty acids in the test diets. The monounsaturated fatty acids were significantly higher in fillet of fish fed diet FO, CSO40 and CSO60 compared to other treatments while saturated and polyunsaturated fatty acids (PUFA) were not affected by the dietary treatment. Some fatty acids (18:0, 18:1n-9, 20:5n-3 and 22:6n-3) were present in higher concentration in fillet lipid than in the CSO100 dietary lipid indicating accumulation in fillet relative to test diets. Retention of n-3 LC-PUFA within the fillet was increasingly inefficient among fish fed increasing levels of FO. Thus, this study suggests that CSO can be considered as a relatively effective substitute for fish oil in European sea bass (35 g) in terms of growth performances and feed efficiency as far as fish meal is present in the diet.

Fatty acid metabolism in European sea bass (Dicentrarchus labrax) : effects of n-6 PUFA and MUFA in fish oil replaced diets

Monounsaturated fatty acids (MUFA)-rich and n-6 polyunsaturated fatty acid (n-6 PUFA)-rich vegetable oils are increasingly used as fish oil replacers for aquafeed formulation. The present study investigated the fatty acid metabolism in juvenile European sea bass (Dicentrarchus labrax, 38.4 g) fed diets containing fish oil (FO, as the control treatment) or two different vegetable oils (the MUFA-rich canola/rapeseed oil, CO, and the n-6 PUFA-rich cottonseed oil, CSO) tested individually or as a 50/50 blend (CO/CSO). The whole-body fatty acid balance method was used to deduce the apparent in vivo fatty acid metabolism. No effect on growth performance and feed utilization was recorded. However, it should be noted that the fish meal content of the experimental diets was relatively high, and thus the requirement for n-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA) may have likely been fulfilled even if dietary fish oil was fully replaced by vegetable oils. Overall, relatively little apparent in vivo fatty acid bioconversion was recorded, whilst the apparent in vivo ?-oxidation of dietary fatty acid was largely affected by the dietary lipid source, with higher rate of ?-oxidation for those fatty acids which were provided in dietary surplus. The deposition of 20:5n-3 and 22:6n-3, as % of the dietary intake, was greatest for the fish fed on the CSO diet. It has been shown that European sea bass seems to be able to efficiently use n-6 PUFA for energy substrate, and this may help in minimizing the ?-oxidation of the health benefiting n-3 LC-PUFA and thus increase their deposition into fish tissues.

Enhancement of gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) larval growth by dietary vitamin E en relation to diffetent levels of essential fatty acids

[EN]Most marine fish larvae require high amounts of n-3 HUFA (highly unsaturated fatty acids) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Watanabe, 1982; Izquierdo, 1996). Fish larvae tissue lipids are also very high in n-3 HUFA, what implies a higher risk of peroxidation (Sargent et al. 1999) and cellular damage (Kanazawa, 1991), requiring then antioxidants to protect them intra- and extra-cellularly from free radical compounds. Vitamin E (Vit E) functions as a chain breaking antioxidant, reacting with the lipid peroxide radical produced and preventing the further reaction with a new PUFA. Hence their requirements are related with the dietary and tissue PUFA contents. The objective of the present study was to determine the effect of dietary Vit E on gilthead sea bream and sea bass survival, growth and stress, at different n-3 HUFA levels.

Effects of dietary mannan oligosaccharides (MOS) on European sea bass (Dicentrarchus labrax) juvenile culture

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Effects of dietary conjugated linoleic acid (CLA) on european sea bass (Dicentrarchus labrax) culture

Programa de Doctorado: Acuicultura

Oxidative stress in sea bass (Dicentrarchus labrax) larvae fed on high DHA microdiets. Involvement of several antioxidant nutrients

Programa de Doctorado, Acuicultura: Producción controlada de organismos acuáticos

Seawater carbonate chemistry and biological parameters during experiments with white sea bass Atractoscion nobilis, 2009

A large fraction of the carbon dioxide added to the atmosphere by human activity enters the sea, causing ocean acidification. We show that otoliths (aragonite ear bones) of young fish grown under high CO2 (low pH) conditions are larger than normal, contrary to expectation. We hypothesize that CO2 moves freely through the epithelium around the otoliths in young fish, accelerating otolith growth while the local pH is controlled. This is the converse of the effect commonly reported for structural biominerals.

European sea bass, Dicentrarchus labrax, in a changing ocean

Ocean acidification, caused by rising concentrations of carbon dioxide (CO2), is widely considered to be a major global threat to marine ecosystems. To investigate the potential effects of ocean acidification on the early life stages of a commercially important fish species, European sea bass (Dicentrarchus labrax), 12 000 larvae were incubated from hatch through metamorphosis under a matrix of two temperatures (17 and 19 °C) and two seawater pCO2 levels (ambient and 1,000 µatm) and sampled regularly for 42 days. Calculated daily mortality was significantly affected by both temperature and pCO2, with both increased temperature and elevated pCO2 associated with lower daily mortality and a significant interaction between these two factors. There was no significant pCO2 effect noted on larval morphology during this period but larvae raised at 19 °C possessed significantly larger eyes and lower carbon:nitrogen ratios at the end of the study compared to those raised under 17 °C. Similarly, when the incubation was continued to post-metamorphic (juvenile) animals (day 67-69), fish raised under a combination of 19 °C and 1000 µatm pCO2 were significantly heavier. However, juvenile D. labrax raised under this combination of 19 °C and 1000 µatm pCO2 also exhibited lower aerobic scopes than those incubated at 19 °C and ambient pCO2. Most studies investigating the effects of near-future oceanic conditions on the early life stages of marine fish have used incubations of relatively short durations and suggested that these animals are resilient to ocean acidification. Whilst the increased survival and growth observed in this study supports this view, we conclude that more work is required to investigate whether the differences in juvenile physiology observed in this study manifest as negative impacts in adult fish.

Combined impacts of elevated CO2 and anthropogenic noise on European sea bass (Dicentrarchus labrax)

Ocean acidification (OA) and anthropogenic noise are both known to cause stress and induce physiological and behavioural changes in fish, with consequences for fitness. OA is also predicted to reduce the ocean's capacity to absorb low-frequency sounds produced by human activity. Consequently, anthropogenic noise could propagate further under an increasingly acidic ocean. For the first time, this study investigated the independent and combined impacts of elevated carbon dioxide (CO2) and anthropogenic noise on the behaviour of a marine fish, the European sea bass (Dicentrarchus labrax). In a fully factorial experiment crossing two CO2 levels (current day and elevated) with two noise conditions (ambient and pile driving), D. labrax were exposed to four CO2/noise treatment combinations: 400 µatm/ambient, 1000 µatm/ambient, 400 µatm/pile-driving, and 1000 µatm/pile driving. Pile-driving noise increased ventilation rate (indicating stress) compared with ambient noise conditions. Elevated CO2 did not alter the ventilation rate response to noise. Furthermore, there was no interaction effect between elevated CO2 and pile-driving noise, suggesting that OA is unlikely to influence startle or ventilatory responses of fish to anthropogenic noise. However, effective management of anthropogenic noise could reduce fish stress, which may improve resilience to future stressors.

Fertilization strategies for Sea Bass Dicentrarchus labrax (Linnaeus, 1758): effects of pre-incubation and duration of egg receptivity in seawater

Studying gamete biology can provide important information about a species fertilization strategy as well as their reproductive ecology. Currently, there is a lack of knowledge about how long sea bass Dicentrarchus labrax eggs can remain viable after being activated in seawater. The objectives of this study were to understand the effects of pre-incubation of fresh and overripe sea bass eggs in seawater and to determine the duration of egg receptivity. Pooled eggs (fresh and overripe) from four females were pre-incubated in seawater for 0 min (control), 0.5 min, 1 min, 3 min, 10 min and 30 min and then fertilized by pooled sperm from four males. The fresh eggs had a higher fertilization success than overripe eggs. Our results revealed a significant effect of pre-incubation time for both the fresh (P < 0.01) and overripe eggs (P < 0.01). Fertilization success of eggs significantly declined for both these treatments after 3 min of pre-incubation, which clearly indicates that sea bass eggs are able to be fertilized by sperm for up to 3 min after release into seawater. This study has particular importance for understanding fertilization strategies, reproductive potential, as well as reproductive ecology of sea bass.

Evaluation of the impact of polyethylene microbeads ingestion in European sea bass (Dicentrarchus labrax) larvae

Microplastics are present in marine habitats worldwide and may be ingested by low trophic organisms such as fish larvae, with uncertain physiological consequences. The present study aims at assessing the impact of polyethylene (PE 10-45µM) microbeads ingestion in European sea bass (Dicentrarchus labrax) larvae. Fish were fed an inert diet including 0, 104 and 105 fluorescent microbeads per gram from 7 until 43 days post-hatching (dph). Microbeads were detected in the gastrointestinal tract in all fish fed diet incorporating PE. Our data revealed an efficient elimination of PE beads from the gut since no fluorescent was observed in the larvae after 48h depuration. While the mortality rate increased significantly with the amount of microbeads scored per larvae at 14 and 20 dph, only ingestion of the highest concentration slightly impacted mortality rates. Larval growth and inflammatory response through Interleukine-1-beta (IL-1) gene expression were not found to be affected while cytochrome-P450-1A1 (cyp1a1) expression level was significantly positively correlated with the number of microbeads scored per larva at 20 dph. Overall, these results suggest that ingestion of PE microbeads had limited impact on sea bass larvae possibly due to their high potential of egestion