Response Of Sea Bass Research Articles

RNA-Seq analysis of juvenile gilthead sea bream (Sparus aurata) provides some clues regarding their resistance to the nodavirus RGNNV genotype

Gilthead sea bream (Sparus aurata) is considered an asymptomatic carrier for the nodavirus genotype affecting European sea bass (Dicentrarchus labrax), RGNNV. Only larvae and juveniles of sea bream have been found to be susceptible to the RGNNV/SJNNV reassortant. Nevertheless, the molecular bases of the high resistance of sea bream against RGNNV are not known, and the overall transcriptome response to the virus remains unexplored. In this work, we conducted the first RNA-Seq analysis of sea bream infected with RGNNV to elucidate the immune mechanisms involved in their resistance. Since we recently published the transcriptome response of sea bass infected with RGNNV, we wanted to take the same tissues (brain and head kidney) at the same time points (24 and 72 h postinfection) to conduct comparative analyses. Sea bream responded to RGNNV challenge with a powerful immune arsenal characterized by the high expression of a multitude of type I interferon-related genes, immune receptors and antigen presentation-related genes in both tissues. Moreover, complement-, coagulation- and angiogenesis-related genes were highly enriched in the head kidney at the earlier sampling point. Interestingly, despite the strong immune response found in the brain, inflammation seems to have been restrained, resulting in a neuroprotective scenario. While the response in sea bass was characterized by the activation of the stress axis, which could lead to immunosuppression and neuronal damage, genes involved in these processes were not modulated in sea bream. An efficient antiviral response accompanied by low inflammation and the absence of stimulation of the stress response seem to play a role in the success of sea bream in resisting RGNNV infection.

Suspended Structures Reduce Variability of Group Risk-Taking Responses of Dicentrarchus labrax Juvenile Reared in Tanks

Structural enrichment is considered a useful tool to improve the welfare conditions of captive fish by deliberately increasing the physical heterogeneity and complexity of captivity environments. However, the potential effects of structural enrichment on the stress response at the group level and on social interactions have not been well studied yet. In this study, we demonstrate that suspended vertical structures (U-shaped ropes) can reduce behavioural variability among fish groups (tank level) of European seabass (Dicentrarchus labrax) juveniles. Differences in behavioural responses during group risk-taking tests (e.g., number of passes per fish) between treatments were detected, and these responses in seabass in enriched captive conditions were more homogeneous among tanks compared to fish from non-enriched tanks. These results suggest a positive effect of the structural enrichment on social stabilisation and response to stressful events at the tank level in seabass. However, further research is still needed to improve the knowledge of the potential effects of structural enrichment on fish welfare and aquaculture management, considering different enrichment designs, intensities, and strategies according to farming conditions, biological needs, and preferences of the fish species and life-stage reared in captivity.

Effects of Melissa officinalis L. Essential Oil in Comparison with Anaesthetics on Gill Tissue Damage, Liver Metabolism and Immune Parameters in Sea Bass (Lateolabrax maculatus) during Simulated Live Transport.

Simple SummaryThe traditional method of transporting live fish in water often involves long driving durations and a high transport density. These conditions can induce stress in the fish, severely effecting their immune system and organs, such as the gills, liver and gut. In this study, Melissa officinalis L. essential oil (MOEO) and two kinds of common anaesthetics were applied to live fish during transport to reduce mortality and stress responses in sea bass (Lateolabrax maculatus). Sea bass were transported for 72 h with a transport density of 250 kg/m3 in different concentrations of MOEO and anaesthetics. The effect of MOEO on tissue damage, energy metabolism and on some oxidative stress and immune parameters of the animals were determined and compared with the effect of the anaesthetics. The results of this study indicated that the energy metabolism, gill and liver tissue damage and immune responses of anesthetized and sedated fish were lower than the control fish. MOEO can reduce the effects of stress and tissue damage in live fish when used as a novel sedative and anaesthetic.In the current study, Melissa officinalis L. essential oil (MOEO), a novel sedative and anaesthetic, was employed in transport water to obtain a lower stress effect and higher survival rate for live marine fish. The effect of MOEO and various types of anaesthetics, administered at a low temperature on gill morphology, liver function and immunological parameters of living sea bass (Lateolabrax maculatus) subjected to transport stress, was evaluated to optimize the anaesthetic and sedative concentrations during live sea bass transport. Light microscopy and scanning electron microscopy of sea bass, subjected to simulated live transport for 72 h, demonstrated that the changes in the morphological characteristics of gill tissue treated with 40 mg/L MOEO (A3 group) were minimal in comparison to those observed in untreated sea bass. The results of pyruvate kinase (PK), phosphofructokinase (PFK), hexokinase (HK), hepatic glycogen (Gly), superoxide dismutase (SOD), lipid peroxides (MDA) and Caspase-3 assays indicated that the glycolysis rate, energy consumption, lipid peroxidation and hepatocyte apoptosis were the lowest in the A3 group. The values of the two immune parameters, lysozyme (LZM) and fish immunoglobulin M (IgM), indicated the strongest immunity ability in the A3 group. After 12 h recovery, sea bass treated with 30 mg/L MS-222 (B group) displayed a 100% survival rate, sea bass treated with 20 mg/L (A2 group) and 40 mg/L (A3 group) MOEO displayed a 96% survival rate, sea bass treated with 20 mg/L eugenol (C group) had a 94% survival rate, and untreated sea bass (CK group) had a 50% survival rate. Therefore, the addition MOEO to the transport water had anaesthetic and sedative effects similar to MS-222 and eugenol. The results confirmed that the addition of MOEO to the transport water could reduce tissue damage, energy metabolism, and the oxidative stress response in sea bass during transport.

Nervous Necrosis Virus-like Particle (VLP) Vaccine Stimulates European Sea Bass Innate and Adaptive Immune Responses and Induces Long-Term Protection against Disease.

The rapidly increasing Mediterranean aquaculture production of European sea bass is compromised by outbreaks of viral nervous necrosis, which can be recurrent and detrimental. In this study, we evaluated the duration of protection and immune response in sea bass given a single dose of a virus-like particle (VLP)-based vaccine. Examinations included experimental challenge with nervous necrosis virus (NNV), serological assays for NNV-specific antibody reactivity, and immune gene expression analysis. VLP-vaccinated fish showed high and superior survival in challenge both 3 and 7.5 months (1800 and 4500 dd) post-vaccination (RPS 87 and 88, OR (surviving) = 16.5 and 31.5, respectively, p < 0.01). Although not providing sterile immunity, VLP vaccination seemed to control the viral infection, as indicated by low prevalence of virus in the VLP-vaccinated survivors. High titers of neutralizing and specific antibodies were produced in VLP-vaccinated fish and persisted for at least ~9 months post-vaccination as well as after challenge. However, failure of immune sera to protect recipient fish in a passive immunization trial suggested that other immune mechanisms were important for protection. Accordingly, gene expression analysis revealed that VLP-vaccination induced a mechanistically broad immune response including upregulation of both innate and adaptive humoral and cellular components (mx, isg12, mhc I, mhc II, igm, and igt). No clinical side effects of the VLP vaccination at either tissue or performance levels were observed. The results altogether suggested the VLP-based vaccine to be suitable for clinical testing under farming conditions.

Cognitive appraisal in fish: stressor predictability modulates the physiological and neurobehavioural stress response in sea bass.

The role of cognitive factors in triggering the stress response is well established in humans and mammals (aka cognitive appraisal theory) but very seldom studied in other vertebrate taxa. Predictability is a key factor of the cognitive evaluation of stimuli. In this study, we tested the effects of stressor predictability on behavioral, physiological and neuromolecular responses in the European sea bass (Dicentrarchus labrax). Groups of four fish were exposed to a predictable (signalled) or unpredictable (unsignalled) stressor. Stressor predictability elicited a lower behavioural response and reduced cortisol levels. Using the expression of immediate early genes (c-fos, egr-1, bdnf and npas4) as markers of neuronal activity, we monitored the activity of three sea bass brain regions known to be implicated in stressor appraisal: the dorsomedian telencephalon, Dm (putative homologue of the pallial amygdala); and the dorsal (Dld) and ventral (Dlv) subareas of the dorsolateral telencephalon (putative homologue of the hippocampus). The activity of both the Dm and Dlv significantly responded to stressor predictability, suggesting an evolutionarily conserved role of these two brain regions in information processing related to stressor appraisal. These results indicate that stressor predictability plays a key role in the activation of the stress response in a teleost fish, hence highlighting the role of cognitive processes in fish stress.

The role of CD8α in the immune response of sea bass against nodavirus

The role of CD8α in the immune response of sea bass against nodavirus

Vaccination of European sea bass Dicentrarchus labrax with avirulent Mycobacterium marinum (iipA::kan mutant)

Mycobacteriosis is a chronic progressive disease affecting teleost fishes all over the world. No vaccine is commercially available against its main etiological agent, Mycobacterium marinum. The mycobacterial gene responsible for invasion and intracellular persistence, iipA, is known to modulate M. marinum pathology. The innate and adaptive immune responses in sea bass (Dicentrarchus labrax) vaccinated with M. marinum iipA::kan mutant with (and without) the use of adjuvant, with (and without) a booster vaccination were monitored. The adjuvanted vaccine induced enhanced immune responses. TNF-α transcription levels were extremely high in spleen of the fish vaccinated with the addition of adjuvant in both fish vaccinated once and twice, followed by an IgM response highly specific for M. marinum. Also, histologically, granulomas started appearing in spleen and head-kidney tissues (but with no visible bacteria) within a month after vaccination, mainly with the adjuvanted vaccine. This was followed by reduction in pathology, as demonstrated by the lower number of granulomas (with visible bacteria), indicating that even heat-killed bacteria were able to elicit granulomatous formations. Adhesion of the internal organs and moderate pigmentation were observed in the perivisceral adipose tissue of nearly all vaccinated fish. Although the adjuvanted heat-killed avirulent iipA::kan mutant clearly induced a strong humoral and adaptive immune response, the booster treatment did not seem to have produced a significantly higher degree of protection from the disease compared to fish that received a single vaccination.

Cross-effects of dietary probiotic supplementation and rearing temperature on growth performance, digestive enzyme activities, cumulative mortality and innate immune response in seabass (Dicentrarchus labrax)

Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross-effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 108 CFU/kg diet), and tested against a non-supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α-amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.

Evolution of Th2 responses: characterization of IL-4/13 in sea bass (Dicentrarchus labrax L.) and studies of expression and biological activity

Th2 immunity is a primary host defence against metazoan pathogens and two of the important cytokines involved in this immune response in mammals are IL-4 and IL-13. Recently the origin and evolution of Th2 immune responses have been investigated in fish where a molecule with relatedness to both IL-4 and IL-13 is present, termed IL-4/13. Different IL-4/13 paralogues (IL-4/13 A and IL-4/13B) exist in teleost fish. In this paper, we have focused on the IL-4/13 isoforms found in the European sea bass (Dicentrarchus labrax L.). Two tandem duplicated but divergent IL-4/13 A isoforms and one IL-4/13B are present, a unique situation compared to other teleosts. These genes were studied in terms of their in vitro and in vivo transcript levels after different treatments and their biological activities after production of the recombinant isoforms. The results show that the presence of these three paralogues is associated with different activities, both in terms of their expression profiles and the ability of the proteins to modulate the expression of immune genes in head kidney leukocytes. It is clear that the initiation and control of type-2 responses in seabass is complex due to the presence of multiple IL-4/13 isoforms with overlapping but distinct activities.

Compensatory growth response of European sea bass (Dicentrarchus labraxL.) under cycled starvation and restricted feeding rate

The compensatory growth response of the European sea bass (Dicentrarchus labrax) that faced cycling starvation and restricted ration was assessed. Juveniles (10.5 g) were stocked into 15 tanks at a density of 25 fish per tank. Five different feeding regimes were tested on triplicate groups of fish: CSatiation: control fed for 60 days without deprivation, CRestricted: 25% restricted feeding, S-R: 1 day starvation then 4 days CRestricted feeding, R-F: 1 day CRestricted feeding then 4 days CSatiation feeding, and, finally, S-F: 1 day starvation then 4 days CSatiation feeding. The specific growth rate of fish in the CSatiation (2.5 ± 0.06% day−1), S-F (2.5 ± 0.11% day−1) and R-F (2.4 ± 0.18% day−1) were significantly higher than that of CRestricted (2.2 ± 0.05% day−1) or S-R (2.0 ± 0.01% day−1). Fish in S-F group were able to achieve catching up with the CSatiation. There was no significant difference in feed conversion rates but R-F and S-F consumed approximately 34% more feed than CSatiation following the first re-feeding day. Although, the highest lipid content was observed in CSatiation (14.4%), S-R (33.3% dry matter) had the highest water content compared with the CSatiation and CRestricted (37.8% and 36.9% dry matter respectively). In conclusion, it may be concluded that sea bass has rapid response to cycling starvation/re-feeding and that a 25% restricted feeding ratio is insufficient to invoke a compensatory growth response in sea bass.

Analysis of the Expression and Modulation of Selected mmunerelated Gene Transcripts in the DLEC Cell Line from European Sea Bass (Dicentrarchus labrax L.)

Cell lines have been established from different fish species especially for virus isolation and for studying cell-pathogen interactions, and therefore are of interest in aquaculture. In this paper, we have investigated the presence and the regulation of some immune genes in the DLEC (Dicentrarchus labrax embryonic cells) cell line from European sea bass (Dicentrarchus labrax L.) to preliminary elucidate their action. The basal expression of the selected genes (interleukin- 1? (IL-1?), cyclooxygenase-2 (COX-2), transforming growth factor-? (TGF-?), CD8-?, major histocompatibility complex II-? (MHC II-?), interferon (IFN) and Mx protein (Mx)) have been investigated and, successively, their modulation have been studied both after stimulation with different mitogen agents and after a transfection with a sequence codifying for the coat protein of a fish nervous necrosis virus (NNV). The results have evidenced that the inflammatory molecules (IL-1?, COX-2, TGF-?), constitutively expressed by the DLEC cell line, are not up-regulated by the stimulation with lipopolysaccharide (LPS) from E. coli, whether the expression of the T-cell marker transcripts (CD8-?, MHC II-?) is influenced by the action of a lectin from Phaseolus vulgaris (PHA-L). Finally, the expression of the coat NNV protein in the DLEC cell line, after the transfection, led to an high up-regulation of IFN and Mx gene transcripts. These data suggest that the DLEC cell line recognize specific pathogen-associated molecular patterns (PAMPs) and, therefore, could be useful for studying T-cell pathways and viral responses in sea bass avoiding the use of live test animals.

Molecular characterisation and structural analysis of an interferon homologue in sea bass ( Dicentrarchus labrax L.)

The interferons (IFNs) are a large family of soluble cytokines involved in the immune response against viral pathogens. Three families of IFNs have been identified in mammals (type I, type II and type III) and, recently, homologues of type I and type II genes have been found in various teleost fish species. In this paper we report the cloning of a cDNA encoding an type I IFN molecule from sea bass ( Dicentrarchus labrax L.), its expression analysis and gene structure and, finally, its 3D structure obtained by template-based modelling. The sea bass IFN cDNA consists of 1047 bp that translates in one reading frame to give the entire molecule containing 185 amino acids. The analysis of the sequence revealed the presence of a putative 22 amino acid signal peptide, two cysteine residues and three potential N-glycosylation sites. The sea bass IFN gene contains four introns as with other type I IFN teleost genes, except medaka that contains three introns. Real time PCR was performed after poly I:C stimulation of DLEC cell line to investigate the expression of sea bass IFN and Mx and an induction was observed for both genes. The predicted 3D structure of sea bass IFN is characterized by an “all-alpha” domain that shows an “up-down bundle” architecture made of six helices (ABB′CDE). The two cysteine residues present in the sequence (i.e. Cys 23 and Cys 126) are in a position and at a distance that suggest the possible formation of a disulfide bridge that may stabilize the structure. Our results will give the opportunity to investigate more in detail antiviral immune responses in sea bass and add to studies on the evolution of the IFN system in teleosts and vertebrates more generally.

Short- and long-term effects of a dietary yeast β-glucan (Macrogard) and alginic acid (Ergosan) preparation on immune response in sea bass ( Dicentrarchus labrax)

The present study investigated the immunomodulatory activity of Ergosan, an algal extract containing alginic acid, and Macrogard, a yeast extract containing β-glucans, on innate and specific immunity in sea bass ( Dicentrarchus labrax). Four cycles of experimental feeding using normal fish feed formulation (control group) supplemented with Ergosan (0.5%) or Macrogard (0.1%) were performed at 60-day intervals (15 days of treatment + 45 days of suspension). Serum complement, lysozyme, total proteins and heat shock protein (HSP) concentrations were measured at 15, 30 and 45 days from the end of the first 15-day feeding cycle (short term) and 45 days after the end of each feeding cycle over a 35-week period (long term). The percentage of B- and T-lymphocytes in peripheral blood leucocytes and gut were measured over long-term trial. Significant elevation ( P < 0.05) in serum complement activity occurred in sea bass fed with alginic acid and glucans, at 15 days from the end of first cycle of treatment. Significant elevation ( P < 0.05) in serum lysozyme, gill and liver HSP concentration were observed in the same experimental groups at 30 days from the end of treatment, whereas a significant increase ( P < 0.05) of complement activity was only observed in fish that received an Ergosan diet. At 45 days from the end of treatment, complement, lysozyme and HSP concentration did not differ among groups. Over the long-term period, no significant differences were observed in innate and specific immune parameters, survival, growth performances and conversion index in treated and control fish. A dramatic decrease of both innate and acquired immune parameters was observed during the winter season in all groups, followed by a partial recovery when water temperature increased. Reduction in complement and lysozyme activities was significatively correlated ( p < 0.01) to water temperature variation. The results suggested the potential of alginic acid and β-glucans to activate some innate immune responses in sea bass, and particularly under conditions of immunodepression related to environmental stress.

Antibody response in sea bass (Dicentrarchus labraxL.) in relation to water temperature and oxygenation

The influence of water temperature and water oxygenation on the specific antibody response was evaluated by indirect ELISA in sea bass (Dicentrarchus labrax L.) immunized against human-gamma-globulins emulsified in Freund's complete adjuvant. A higher antibody response was observed in fish reared at 24 and 30 degrees C, than at 12 and 18 degrees C. For the fish group reared at 24 degrees C, the immunological observation was carried out until 300 days after immunization. At the end of this period, fish kept at 24 degrees C still showed immunological competence, although the antibody response began to decrease significantly from 120 days after the immunization. Fish reared under mild hyperoxia and normoxia conditions had higher antibody responses than fish reared under mild hypoxia conditions. Moreover, at the end of the experimental period of 56 days the antibody response of fish reared under hyperoxia conditions was significantly higher than the immune response detected in fish reared under normoxia and hypoxia conditions.